Science.gov

Sample records for ca tu thy

  1. 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

  2. The Thy Project. Final Report.

    ERIC Educational Resources Information Center

    Billimoria, Roshan R., Ed.

    Inspiried by a United Nations effort to establish a worldwide university, the four and one-half year project carried out in Thy (Denmark) is explained in this final report, from its historical beginnings in 1973 to its official completion in 1978. Dedicated to the solution of problems which could be considered universal, the project goals are…

  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. 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

  6. 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

  7. 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

  8. 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

  9. 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

  10. THI1, a protein involved in the biosynthesis of thiamin in Arabidopsis thaliana: structural analysis of THI1(A140V) mutant.

    PubMed

    Garcia, Assuero F; Dyszy, Fabio; Munte, Claudia E; Demarco, Ricardo; Beltramini, Leila M; Oliva, Glaucius; Costa-Filho, Antonio J; Araujo, Ana P U

    2014-06-01

    In eukaryotes, there are still steps of the vitamin B1 biosynthetic pathway not completely understood. In Arabidopsis thaliana, THI1 protein has been associated with the synthesis of the thiazole ring, a finding supported by the identification of a thiamine pyrophosphate (TPP)-like compound in its structure. Here, we investigated THI1 and its mutant THI1(A140V), responsible for the thiamin auxotrophy in a A. thaliana mutant line, aiming to clarify the impact of this mutation in the stability and activity of THI1. Recently, the THI1 orthologue (THI4) was revealed to be responsible for the donation of the sulfur atom from a cysteine residue to the thiazole ring in the thiamine intermediate. In this context, we carried out a cysteine quantification in THI1 and THI1(A140V) using electron spin resonance (ESR). These data showed that THI1(A140V) contains more sulfur-containing cysteines than THI1, indicating that the function as a sulfur donor is conserved, but the rate of donation reaction is somehow affected. Also, the bound compounds were isolated from both proteins and are present in different amounts in each protein. Unfolding studies presented differences in melting temperatures and also in the concentration of guanidine at which half of the protein unfolds, thus showing that THI1(A140V) has its conformational stability affected by the mutation. Hence, despite keeping its function in the early steps during the synthesis of TPP precursor, our studies have shown a decrease in the THI1(A140V) stability, which might be slowing down the biological activity of the mutant, and thus contributing to thiamin auxotrophy.

  11. 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

  12. Isolation and characterization of a thiamin transport gene, THI10, from Saccharomyces cerevisiae.

    PubMed

    Enjo, F; Nosaka, K; Ogata, M; Iwashima, A; Nishimura, H

    1997-08-01

    We isolated a thiamin transporter gene, THI10, from a genomic library of Saccharomyces cerevisiae by the complementation of a yeast mutant defective in thiamin transport activity. The THI10 gene contained an open reading frame of 1,794 base pairs encoding a 598-amino acid polypeptide with a calculated molecular weight of 66, 903. The nucleotide sequence of THI10 is completely identical to that of an anonymous DNA (open reading frame L8083.2) mapped to chromosome XII; two other genes (open reading frames YOR071c and YOR192c) in chromosome XV are extremely similar to THI10. Moreover, the THI10 gene product showed significant sequence homology with yeast allantoin and uracil transporters. Hydropathy profile suggested that THI10 product is highly hydrophobic and contains many transmembrane regions. Gene disruption of the THI10 locus completely abolished the thiamin transport activity and thiamin binding activity in yeast plasma membrane fraction. Both the transport and thiamin binding activities were restored in the disrupted cells when the THI10 open reading frame was expressed by yeast GAL1 promoter, suggesting that the THI10 gene encodes for the thiamin transport carrier protein. Northern blot analysis demonstrated that THI10 gene expression is regulated at the mRNA level by intracellular thiamin pyrophosphate and that it requires a positive regulatory factor encoded by THI3 gene.

  13. 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

  14. Anti-proliferative actions of T-type calcium channel inhibition in Thy1 nephritis.

    PubMed

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

    2013-08-01

    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 Ca(2+) channels (TCCs). The current study investigates the effect of TH1177 on MC proliferation in vitro and in vivo. The effect of Ca(2+) 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

  15. Renaturation of rhodanese by translational elongation factor (EF) Tu. Protein refolding by EF-Tu flexing.

    PubMed

    Kudlicki, W; Coffman, A; Kramer, G; Hardesty, B

    1997-12-19

    The translation elongation factor (EF) Tu has chaperone-like capacity to promote renaturation of denatured rhodanese. This renaturation activity is greatly increased under conditions in which the factor can oscillate between the open and closed conformations that are induced by GDP and GTP, respectively. Oscillation occurs during GTP hydrolysis and subsequent replacement of GDP by EF-Ts which is then displaced by GTP. Renaturation of rhodanese and GTP hydrolysis by EF-Tu are greatly enhanced by the guanine nucleotide exchange factor EF-Ts. However, renaturation is reduced under conditions that stabilize EF-Tu in either the open or closed conformation. Both GDP and the nonhydrolyzable analog of GTP, GMP-PCP, inhibit renaturation. Kirromycin and pulvomycin, antibiotics that specifically bind to EF-Tu and inhibit its activity in peptide elongation, also strongly inhibit EF-Tu-mediated renaturation of denatured rhodanese to levels near those observed for spontaneous, unassisted refolding. Kirromycin locks EF-Tu in the open conformation in the presence of either GTP or GDP, whereas pulvomycin locks the factor in the closed conformation. The results lead to the conclusion that flexing of EF-Tu, especially as occurs between its open and closed conformations, is a major factor in its chaperone-like refolding activity.

  16. 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

  17. Analysis of ThiC variants in the context of the metabolic network of Salmonella enterica.

    PubMed

    Palmer, Lauren D; Dougherty, Michael J; Downs, Diana M

    2012-11-01

    In bacteria, the 4-amino-hydroxymethyl-2-methylpyrimidine (HMP) moiety of thiamine is synthesized from 5-aminoimidazole ribotide (AIR), a branch point metabolite of purine and thiamine biosynthesis. ThiC is a member of the radical S-adenosylmethionine (AdoMet) superfamily and catalyzes the complex chemical rearrangement of AIR to HMP-P. As reconstituted in vitro, the ThiC reaction requires AdoMet, AIR, and reductant. This study analyzed variants of ThiC in vivo and in vitro to probe the metabolic network surrounding AIR in Salmonella enterica. Several variants of ThiC that required metabolic perturbations to function in vivo were biochemically characterized in vitro. Results presented herein indicate that the subtleties of the metabolic network have not been captured in the current reconstitution of the ThiC reaction.

  18. THY1 as a reliable marker for enrichment of undifferentiated spermatogonia in the goat.

    PubMed

    Abbasi, Hassan; Tahmoorespur, Mojtaba; Hosseini, Sayyed Morteza; Nasiri, Zahra; Bahadorani, Mehrnoosh; Hajian, Mahdi; Nasiri, Mohammad Reza; Nasr-Esfahani, Mohammad Hossein

    2013-11-01

    Spermatogonial stem cells are unique cells of testes that can restore fertility upon transplantation into recipient testes. However, use of suitable markers for enrichment of these cells have important potential application. THY1, is an established conserved marker of spermatogonial stem cells in bovine, rodents, and primates, but there is no information available in goats. After three rounds of enzymatic digestion of prepubertal goat testicular tissues, undifferentiated spermatogonia positive for THY1 were isolated by magnetic-activated cell sorting and were used for immunocytochemistry, real-time polymerase chain reaction analysis for gene expression, protein expression, and transplantation into recipient mice. Immunocytochemical analyses showed that significantly higher percentage of THY1(+) cells were positive for PLZF and VASA when compared with unselected population. This result for PLZF was further confirmed at the protein level. Real-time polymerase chain reaction analysis revealed that expression of THY1, PLZF, VASA, BCL6B, and UCHL1 as SCCs characteristic genes in THY1(+) cells was significantly higher than in the initial population. Finally, transplantation of PKH26-labeled cells revealed that THY1(+) cells had higher capacity for colony formation when compared with unselected cells. In conclusion, the results provide indications that THY1 surface marker can be reliably used for enrichment of undifferentiated spermatogonial in the goats.

  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. Domain organization in Candida glabrata THI6, a bifunctional enzyme required for thiamin biosynthesis in eukaryotes.

    PubMed

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

    2010-11-16

    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-5β-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.

  2. Luminescence dating of Holocene aeolian sand movement, Thy, Denmark

    NASA Astrophysics Data System (ADS)

    Murray, A. S.; Clemmensen, L. B.

    2001-12-01

    We report on the application of optically stimulated luminescence dating to an investigation of the frequency of periods of aeolian sand movement and dune formation. On the coast of Thy, Holocene aeolian sediments, interleaved with peaty palaeosols, form some of the most complete successions known in western Denmark. Samples were taken from a coastal exposure, and an inland archaeological site. Dosimetry was based on laboratory gamma spectrometry, and quartz luminescence measurements used the single-aliquot regenerative-dose protocol, and 470±30 nm stimulation. The sand samples cover an age ranging from about 100 years to 4500 years; the youngest dates give us confidence that the material was well bleached at deposition. Good agreement was obtained with the radiocarbon palaeosol ages. We identify three main periods of aeolian sand movement, starting at about 4200, 2700 and 900 years ago; at least during the latter two periods significant aeolian sand movement occurred. All three phases of aeolian activity were separated by long periods of stability and soil formation. A more recent episode of dune formation, restricted to the coast and beginning less than 200 years ago, may have been triggered by modern coastal erosion.

  3. 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...

  4. 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...

  5. 40 CFR 264.553 - Temporary Units (TU).

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... following factors: (1) Length of time such unit will be in operation; (2) Type of unit; (3) Volumes of... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Temporary Units (TU). 264.553 Section... Provisions for Cleanup § 264.553 Temporary Units (TU). (a) For temporary tanks and container storage...

  6. On the localization of Thy-1-like immunoreactivity in the rodent and human nervous system.

    PubMed

    Seiger, A; Almqvist, P; Granholm, A C; Olson, L

    1986-01-01

    The neuronal localisation of the surface glycoprotein Thy-1 was studied using the adult mouse iris whole-mount preparation. Polyclonal antibodies to Thy-1 and indirect immunohistochemical techniques were used on fixed tissues. In the adult intact mouse iris a plexus of delicate bundles and fibres was found in both the dilator and sphincter regions. Ovoid negative spots along the bundles were numerous, probably indicating the location of supportive cells. The ciliary body contained strongly immunoreactive bundles oriented in radial and circular patterns. Numerous Thy-1-positive mast cells were found in the irides. All Thy-1-immunoreactive fibres disappeared in intraocular iris transplants after 4 days, leaving only the Thy-1-positive mast cells. A Thy-1-positive fibre plexus reappeared in intraocular iris transplants after 4 weeks, strongly indicating that Thy-1-immunoreactive fibres in adult mouse irides are associated with the nerve fibres and not with their supportive tissue. Distribution of Thy-1-like immunoreactivity in the developing human nervous system is presented for the first time, and its temporal changes are followed from the eighth gestational week to adulthood. At eight weeks the spinal cord and lower brain stem seemed to show virtually no immunoreactivity. At 10 and 31 weeks gestational age immunoreactivity was found preferentially in white matter areas with a granular appearance, becoming more densely aggregated at the later stage. Two months postnatally the internal capsule was strongly positive in an otherwise negative neuropil, only to disappear completely in the adult brain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2875228

  7. 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

  8. 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

  9. 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.

  10. 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

  11. Molecular characterization of a thiJ-like gene in Chinese cabbage.

    PubMed

    Oh, Kyung-Jin; Park, Yong-Soon; Lee, Kyung-Ah; Chung, Yong Je; Cho, Tae-Ju

    2004-05-31

    A cDNA clone for a salicylic acid-induced gene in Chinese cabbage (Brassica rapa subsp. pekinensis) was isolated and characterized. The cabbage gene encoding a protein of 392 amino acids contained a tandem array of two thiJ-like sequences. ThiJ is a thiamin biosynthesis enzyme that catalyzes the phosphorylation of hydroxymethylpyrimidine (HMP) to HMP monophosphate. Although the cabbage gene shows a similarity to bacterial thiJ genes, it also shares a similarity with the human DJ-1, a multifunctional protein that is involved in transcription regulation, male fertility, and parkinsonism. The cabbage thiJ-like gene is strongly induced by salicylic acid and a nonhost pathogen, Pseudomonas syringae pv. tomato, which elicits a hypersensitive response in Chinese cabbage. Treatment of the cabbage leaves with BTH, methyl jasmonate, or ethephon showed that the cabbage thiJ-like gene expression is also strongly induced by BTH, but not by methyl jasmonate or ethylene. This indicates that the cabbage gene is activated via a salicylic acid-dependent signaling pathway. Examination of the tissue-specific expression revealed that the induction of the cabbage gene expression by BTH occurs in the leaf, stem, and floral tissues but not in the root.

  12. Generation and Screening of Transgenic Mice with Neuronal Labeling Controlled by Thy1 Regulatory Elements.

    PubMed

    Marinković, Petar; Godinho, Leanne; Misgeld, Thomas

    2015-10-01

    Major progress has been made using in vivo imaging in mice to study mammalian nervous system development, plasticity, and disease. This progress has depended in part on the wide availability of two-photon microscopy, which is capable of penetrating deep into scattering tissue. Equally important, however, is the generation of suitable transgenic mouse models, which provide a "Golgi staining"-like labeling of neurons that is sparse and bright enough for in vivo imaging. Particularly prominent among such transgenic mice are the so-called Thy1-XFP mice (in which XFP stands for any fluorescent protein) that are used in numerous studies, especially to visualize spine plasticity in the cortex and remodeling in peripheral synapses. New generations of Thy1-XFP mice are now being generated at a high rate, and these have allowed previously difficult experiments to become feasible. Moreover, with easy access to core facilities or commercial providers of pronuclear injections, generating simple Thy1 transgenic mice is now a possibility even for small laboratories. In this introduction, we discuss the Thy1 regulatory elements used to generate transgenic lines with neuronal labeling. We provide a brief overview of currently available Thy1 transgenic mice, including lines labeling neuronal organelles or reporting neuronal function. PMID:26430261

  13. Dynamic catch of a Thy-1-α5β1+syndecan-4 trimolecular complex

    NASA Astrophysics Data System (ADS)

    Fiore, Vincent F.; Ju, Lining; Chen, Yunfeng; Zhu, Cheng; Barker, Thomas H.

    2014-09-01

    Cancer cell adhesion to the vascular endothelium is a critical step of tumour metastasis. Endothelial surface molecule Thy-1 (CD90) is implicated in the metastatic process through its interactions with integrins and syndecans. However, how Thy-1 supports cell-cell adhesion in a dynamic mechanical environment is not known. Here we show that Thy-1 supports β1 integrin- and syndecan-4 (Syn4)-mediated contractility-dependent mechanosignalling of melanoma cells. At the single-molecule level, Thy-1 is capable of independently binding α5β1 integrin and syndecan-4 (Syn4) receptors. However, in the presence of both α5β1 and Syn4, the two receptors bind cooperatively to Thy-1, to form a trimolecular complex. This trimolecular complex displays a unique phenomenon we coin ‘dynamic catch’, characterized by abrupt bond stiffening followed by the formation of catch bonds, where force prolongs the bond lifetime. Thus, we reveal a new class of trimolecular interactions where force strengthens the synergistic binding of two co-receptors and modulates downstream mechanosignalling.

  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... New Uses for Specific Chemical Substances § 721.600 3-Alkyl-2-(2-anilino)vinyl thi-azolinium salt... substance identified generically as 3-alkyl-2-(2-ani-lino)vin-yl-thi-azo-li-nium salt (PMN P-84-1007)...

  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... New Uses for Specific Chemical Substances § 721.600 3-Alkyl-2-(2-anilino)vinyl thi-azolinium salt... substance identified generically as 3-alkyl-2-(2-ani-lino)vin-yl-thi-azo-li-nium salt (PMN P-84-1007)...

  16. 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... New Uses for Specific Chemical Substances § 721.600 3-Alkyl-2-(2-anilino)vinyl thi-azolinium salt... substance identified generically as 3-alkyl-2-(2-ani-lino)vin-yl-thi-azo-li-nium salt (PMN P-84-1007)...

  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... New Uses for Specific Chemical Substances § 721.600 3-Alkyl-2-(2-anilino)vinyl thi-azolinium salt... substance identified generically as 3-alkyl-2-(2-ani-lino)vin-yl-thi-azo-li-nium salt (PMN P-84-1007)...

  18. 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... New Uses for Specific Chemical Substances § 721.600 3-Alkyl-2-(2-anilino)vinyl thi-azolinium salt... substance identified generically as 3-alkyl-2-(2-ani-lino)vin-yl-thi-azo-li-nium salt (PMN P-84-1007)...

  19. T helper cells in murine germinal centers are antigen-specific emigrants that downregulate Thy-1

    PubMed Central

    1996-01-01

    After immunization, activated splenic T cells proliferate in periarteriolar lymphoid sheaths (PALS) and subsequently migrate to the lymphoid follicle where they enter nascent germinal centers. Analysis of TCR V(D)J gene rearrangements indicates extensive emigration, frequently involving more than a single white pulp region. These migrants constitute a unique set of T helper cells that express antigen- specific alpha beta TCR, CD3, and CD4, but little or no Thy-1, a differentiation antigen present on the great majority of peripheral murine T lymphocytes. The origin of CD4+ Thy-1 follicular T cells appears to be the Thy+ population in the PALS, as both sets commonly share identical V(D)J rearrangements. PMID:9064325

  20. Expression of thiamin biosynthetic genes (thiCOGE) and production of symbiotic terminal oxidase cbb3 in Rhizobium etli.

    PubMed Central

    Miranda-Ríos, J; Morera, C; Taboada, H; Dávalos, A; Encarnación, S; Mora, J; Soberón, M

    1997-01-01

    In this paper we report the cloning and sequence analysis of four genes, located on plasmid pb, which are involved in the synthesis of thiamin in Rhizobium etli (thiC, thiO, thiG, and thiE). Two precursors, 4-methyl-5-(beta-hydroxyethyl)thiazole monophosphate and 4-amino-5-hydroxymethylpyrimidine pyrophosphate, are coupled to form thiamin monophosphate, which is then phosphorylated to make thiamin pyrophosphate. The first open reading frame (ORF) product, of 610 residues, has significant homology (69% identity) with the product of thiC from Escherichia coli, which is involved in the synthesis of hydroxymethylpyrimidine. The second ORF product, of 327 residues, is the product of a novel gene denoted thiO. A protein motif involved in flavin adenine dinucleotide binding was found in the amino-terminal part of ThiO; also, residues involved in the catalytic site of D-amino acid oxidases are conserved in ThiO, suggesting that it catalyzes the oxidative deamination of some intermediate of thiamin biosynthesis. The third ORF product, of 323 residues, has significant homology (38% identity) with ThiG from E. coli, which is involved in the synthesis of the thiazole. The fourth ORF product, of 204 residues, has significant homology (47% identity) with the product of thiE from E. coli, which is involved in the condensation of hydroxymethylpyrimidine and thiazole. Strain CFN037 is an R. etli mutant induced by a single Tn5mob insertion in the promoter region of the thiCOGE gene cluster. The Tn5mob insertion in CFN037 occurred within a 39-bp region which is highly conserved in all of the thiC promoters analyzed and promotes constitutive expression of thiC. Primer extension analysis showed that thiC transcription in strain CFN037 originates within the Tn5 element. Analysis of c-type protein content and expression of the fixNOQP operon, which codes for the symbiotic terminal oxidase cbb3, revealed that CFN037 produces the cbb3 terminal oxidase. These data show a direct relationship

  1. Thy-1 Regulates VEGF-Mediated Choroidal Endothelial Cell Activation and Migration: Implications in Neovascular Age-Related Macular Degeneration

    PubMed Central

    Wang, Haibo; Han, Xiaokun; Kunz, Eric; Hartnett, M. Elizabeth

    2016-01-01

    Purpose This study addresses the hypothesis that age-related stresses upregulate Thy-1 in choroidal endothelial cells (CECs) and contribute to CEC activation and migration, processes important in choroidal neovascularization (CNV). Methods Measurements were made of Thy-1 protein (Western blot) in CECs and Thy-1 mRNA (real time quantitative PCR) in CECs treated with VEGF, CCL11, or PBS or in RPE/choroids from young or old donors or lasered or nonlasered mice. Immunolabeled Thy-1 in ocular sections was compared from young versus old human donor eyes or those with or without neovascular AMD or from lasered versus nonlasered mice. Choroidal endothelial cells transfected with Thy-1 or control siRNA or pretreated with Thy-1 blocking peptide or control were stimulated with VEGF or 7-ketocholesterol (7-KC). Choroidal endothelial cell migration, proliferation, cytoskeletal stress fibers, Rac1 activation, and phosphorylated VEGF receptor 2 (VEGFR2), integrin β3, and Src were measured. Statistics were performed using ANOVA. Results Thy-1 was expressed in retinal ganglion cells and in vascular endothelial-cadherin–labeled choroid and localized to human or mouse laser-induced CNV lesions. Thy-1 protein and mRNA were significantly increased in CECs treated with VEGF or CCL11 and in RPE/choroids from aged versus young donor eyes or from lasered mice versus nonlasered controls. Knockdown or inhibition of Thy-1 in CECs significantly reduced VEGF-induced CEC migration and proliferation, stress fiber formation and VEGFR2, Src, integrin β3 and Rac1 activation, and 7-KC–induced Rac1 and Src activation. Conclusions Thy-1 in CECs regulates VEGF-induced CEC activation and migration and links extracellular 7-KC to intracellular signaling. Future studies elucidating Thy-1 mechanisms in neovascular AMD are warranted. PMID:27768790

  2. 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

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

    PubMed

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

    2015-10-30

    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.

  5. Engineering the elongation factor Tu for efficient selenoprotein synthesis

    PubMed Central

    Haruna, Ken-ichi; Alkazemi, Muhammad H.; Liu, Yuchen; Söll, Dieter; Englert, Markus

    2014-01-01

    Selenocysteine (Sec) is naturally co-translationally incorporated into proteins by recoding the UGA opal codon with a specialized elongation factor (SelB in bacteria) and an RNA structural signal (SECIS element). We have recently developed a SECIS-free selenoprotein synthesis system that site-specifically—using the UAG amber codon—inserts Sec depending on the elongation factor Tu (EF-Tu). Here, we describe the engineering of EF-Tu for improved selenoprotein synthesis. A Sec-specific selection system was established by expression of human protein O6-alkylguanine-DNA alkyltransferase (hAGT), in which the active site cysteine codon has been replaced by the UAG amber codon. The formed hAGT selenoprotein repairs the DNA damage caused by the methylating agent N-methyl-N′-nitro-N-nitrosoguanidine, and thereby enables Escherichia coli to grow in the presence of this mutagen. An EF-Tu library was created in which codons specifying the amino acid binding pocket were randomized. Selection was carried out for enhanced Sec incorporation into hAGT; the resulting EF-Tu variants contained highly conserved amino acid changes within members of the library. The improved UTu-system with EF-Sel1 raises the efficiency of UAG-specific Sec incorporation to >90%, and also doubles the yield of selenoprotein production. PMID:25064855

  6. Support studies for installing the phosphodiester residues of the Thy-1 glycoprotein membrane anchor.

    PubMed

    Campbell, A S; Fraser-Reid, B

    1994-11-01

    Support studies for late-stage installation of the three different types of phosphodiesters found in the rat brain Thy-1 glycoprotein membrane anchor are described. The strategy is geared towards optimizing convergency and the development of chemoselective procedures including deprotection, phosphorylation, esterification and cysteinylation has been investigated. Some of these procedures are being designed for oligosaccharides containing several unprotected hydroxy groups.

  7. Molecular characterization of Thy1 expressing fear-inhibiting neurons within the basolateral amygdala

    PubMed Central

    McCullough, Kenneth M.; Choi, Dennis; Guo, Jidong; Zimmerman, Kelsey; Walton, Jordan; Rainnie, Donald G.; Ressler, Kerry J.

    2016-01-01

    Molecular characterization of neuron populations, particularly those controlling threat responses, is essential for understanding the cellular basis of behaviour and identifying pharmacological agents acting selectively on fear-controlling circuitry. Here we demonstrate a comprehensive workflow for identification of pharmacologically tractable markers of behaviourally characterized cell populations. Thy1-eNpHR-, Thy1-Cre- and Thy1-eYFP-labelled neurons of the BLA consistently act as fear inhibiting or ‘Fear-Off' neurons during behaviour. We use cell-type-specific optogenetics and chemogenetics (DREADDs) to modulate activity in this population during behaviour to block or enhance fear extinction. Dissociated Thy1-eYFP neurons are isolated using FACS. RNA sequencing identifies genes strongly upregulated in RNA of this population, including Ntsr2, Dkk3, Rspo2 and Wnt7a. Pharmacological manipulation of neurotensin receptor 2 confirms behavioural effects observed in optogenetic and chemogenetic experiments. These experiments identify and validate Ntsr2-expressing neurons within the BLA, as a putative ‘Fear-Off' population. PMID:27767183

  8. 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

  9. 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

  10. 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…

  11. 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.

  12. Biochemical characterization of ThiT from Lactococcus lactis: a thiamin transporter with picomolar substrate binding affinity.

    PubMed

    Erkens, Guus B; Slotboom, Dirk Jan

    2010-04-13

    The putative thiamin transporter ThiT from Lactococcus lactis was overproduced in the membrane of lactococcal cells. In vivo transport assays using radiolabeled thiamin demonstrated that ThiT indeed was involved in thiamin transport. The protein was solubilized from the membranes and purified in detergent solution. Size exclusion chromatography coupled to static light scattering, refractive index, and UV absorbance measurements (SEC-MALLS) showed that ThiT is a monomer of 22.7 kDa in detergent solution. When the cells overexpressing ThiT had been cultivated in complex growth medium, all binding sites of the purified protein were occupied with substrate, which had copurified with the protein. MALDI-TOF mass spectrometry analysis confirmed that the copurified substance was thiamin. Substrate-depleted ThiT was obtained by expressing the protein in cells that were cultivated in chemically defined growth medium without thiamin. The intrinsic tryptophan fluorescence of substrate-depleted ThiT was strongly quenched upon thiamin binding. The quenching of the fluorescence was used to determine dissociation constants for thiamin and related compounds. ThiT had an unusually high affinity for thiamin (K(D) = 122 +/- 13 pM) and bound the substrate with a 1:1 (protein:ligand) stoichiometry. TPP, TMP, and pyrithiamin bound to ThiT with nanomolar affinity. A multiple sequence alignment of ThiT homologues revealed that well-conserved residues were clustered in a tryptophan-rich stretch comprising the loop between the predicted membrane spanning segments 5 and 6. Mutational analysis of the conserved residues in this region combined with binding assays of thiamin and related compounds was used to build a model of the high-affinity binding site. The model was compared with thiamin binding sites of other proteins and interpreted in terms of the transport mechanism.

  13. 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…

  14. 40 CFR 264.553 - Temporary Units (TU).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... or RCRA 3008(h), or at a permitted facility that is not subject to § 264.101, the Regional... 40 Protection of Environment 26 2011-07-01 2011-07-01 false Temporary Units (TU). 264.553 Section 264.553 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES...

  15. Purified CD34+ Lin- Thy+ stem cells do not contain clonal myeloma cells.

    PubMed

    Gazitt, Y; Reading, C C; Hoffman, R; Wickrema, A; Vesole, D H; Jagannath, S; Condino, J; Lee, B; Barlogie, B; Tricot, G

    1995-07-01

    High-dose therapy with autologous marrow or peripheral blood stem cell (PBSC) rescue has been extensively applied in the treatment of multiple myeloma (MM) patients during the past 10 years resulting in improved event-free and overall survival when compared with standard chemotherapy. However, relapses are common and cure is unlikely in the majority of patients. Because both bone marrow and PBSCs are contaminated with myeloma cells it is conceivable that relapse after autotransplantation originates at least in part from autografted tumor cells. In this study, mobilized PBSCs were examined for the presence of myeloma cells based on immunophenotyping and sensitive polymerase chain reaction (PCR)-based techniques. In addition, CD34+ Lin- Thy+ stem cells were purified from mobilized PBSC harvests of 10 MM patients by sequentially using counterflow elutriation centrifugation, treatment with phenylalanine methylester, and flow sorting, using 5-parameter gating (propidium iodide, forward scatter, side scatter, CD34+ v Lin- and CD34+ v Thy+). Virtually all mobilized unsorted PBSC preparations contained myeloma cells in sufficient quantities (range, < 0.01 to > 10%) potentially causing a disease relapse. Stem cell purification led to an overall enrichment by about 50-fold in all 10 patients; approximately 90% of the final cell population expressed CD34+ Lin- Thy+ with no evidence of myeloma cell contamination based on flow cytometric analysis of CD38bright cells (< 0.1%). Quantitative PCR amplification of patient-specific complementarity determining region III (CDRIII) DNA sequences showed depletion of clonal B cells by 2.7 to 7.3 logs, with the highest log reduction noted in the samples initially containing the most tumor cells. Our results show that purification of CD34+ Lin- Thy+ cells depletes myeloma cells to undetectable levels from up to 10% present in unsorted PBSCs, thus offering a tool to investigate whether MM relapse after autotransplantation can be reduced

  16. 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

  17. 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

  18. 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

  19. Purification, characterization, and sequencing of novel antimicrobial peptides, Tu-AMP 1 and Tu-AMP 2, from bulbs of tulip (Tulipa gesneriana L.).

    PubMed

    Fujimura, Masatoshi; Ideguchi, Mineo; Minami, Yuji; Watanabe, Keiichi; Tadera, Kenjiro

    2004-03-01

    Novel antimicrobial peptides (AMP), designated Tu-AMP 1 and Tu-AMP 2, were purified from the bulbs of tulip (Tulipa gesneriana L.) by chitin affinity chromatography and reverse-phase high-performance liquid chromatography (HPLC). They bind to chitin in a reversible way. They were basic peptides having isoelectric points of over 12. Tu-AMP 1 and Tu-AMP 2 had molecular masses of 4,988 Da and 5,006 Da on MALDI-TOF MS analysis, and their extinction coefficients of 1% aqueous solutions at 280 nm were 3.3 and 3.4, respectively. Half of all amino acid residues of Tu-AMP 1 and Tu-AMP 2 were occupied by cysteine, arginine, lysine, and proline. The concentrations of peptides required for 50% inhibition (IC(50)) of the growth of plant pathogenic bacteria and fungi were 2 to 20 microg/ml. The structural characteristics of Tu-AMP 1 and Tu-AMP 2 indicated that they were novel thionin-like antimicrobial peptides, though Tu-AMP 2 was a heterodimer composes of two short peptides joined with disulfide bonds.

  20. Thionin Thi2.1 from Arabidopsis thaliana expressed in endothelial cells shows antibacterial, antifungal and cytotoxic activity.

    PubMed

    Loeza-Angeles, Heber; Sagrero-Cisneros, Eduardo; Lara-Zárate, Leticia; Villagómez-Gómez, Erik; López-Meza, Joel E; Ochoa-Zarzosa, Alejandra

    2008-10-01

    Thionins are plant antimicrobial peptides with antibacterial and antifungal activities. Thionin Thi2.1 cDNA from Arabidopsis thaliana was expressed in BVE-E6E7 bovine endothelial cell line and its activity was evaluated against Escherichia coli, Staphylococcus aureus, Candida albicans and different mammal cell lines. Total protein (2.5 microg) from conditioned medium (CM) of clone EC-Thi2.1 inhibited the growth of E. coli, S. aureus (>90%) and C. albicans strains (>80%) in relation to the CM from control cells. Also, CM of EC-Thi2.1 inhibited the viability of several transformed and normal mammal cell lines (38-95%). These results suggest that thionin Thi2.1 is an antimicrobial peptide that could be use in the treatment of mammalian infectious diseases. PMID:18563581

  1. Problem-Centered Design and Personal Teaching Style: An Exploratory Study of Youguang Tu's Course on Philosophy of Education

    ERIC Educational Resources Information Center

    Lei, Hongde

    2016-01-01

    Youguang Tu is a contemporary Chinese philosopher of education. His course on philosophy of education had a significant impact on his students. This exploratory study examines how Tu designed and taught this course. Ultimately, there are two reasons why Tu's course had such a significant influence on his students. The first is that Tu used…

  2. 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

  3. Thymus cell antigen 1 (Thy1, CD90) is expressed by lymphatic vessels and mediates cell adhesion to lymphatic endothelium.

    PubMed

    Jurisic, Giorgia; Iolyeva, Maria; Proulx, Steven T; Halin, Cornelia; Detmar, Michael

    2010-10-15

    The lymphatic vascular system plays an important role in inflammation and cancer progression, although the molecular mechanisms involved are poorly understood. As determined by comparative transcriptional profiling studies of ex vivo isolated mouse intestinal lymphatic endothelial cells versus blood vascular endothelial cells, thymus cell antigen 1 (Thy1, CD90) was expressed at much higher levels in lymphatic endothelial cells than in blood vascular endothelial cells. These findings were confirmed by quantitative PCR, and at the protein level by FACS and immunofluorescence analyses. Thy1 was also strongly expressed by tumor-associated lymphatic vessels, as evaluated in a B16 melanoma footpad model in mice. Blockade of Thy1 inhibited tumor cell adhesion to cultured mouse lymphatic endothelial cells. Importantly, treatment of human dermal microvascular endothelial cells with tumor necrosis factor or phorbol 12-myristate 13-acetate resulted in Thy1 upregulation in podoplanin-expressing lymphatic endothelial cells, but not in podoplanin-negative blood vascular endothelial cells. Moreover, adhesion of human polymorphonuclear and mononuclear leukocytes to human lymphatic endothelial cells was Thy1-dependent. Together, these results identify Thy1 as a novel lymphatic vessel expressed gene and suggest its potential role in the cell adhesion processes required for tumor progression and inflammation.

  4. 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...

  5. TU elements: a heterogeneous family of modularly structured eucaryotic transposons.

    PubMed Central

    Hoffman-Liebermann, B; Liebermann, D; Kedes, L H; Cohen, S N

    1985-01-01

    We describe here a family of foldback transposons found in the genome of the higher eucaryote, the sea urchin Strongylocentrotus purpuratus. Two major classes of TU elements have been identified by analysis of genomic DNA and TU element clones. One class consists of largely similar elements with long terminal inverted repeats (IVRs) containing outer and inner domains and sharing a common middle segment that can undergo deletions. Some of these elements contain insertions. The second class is highly heterogeneous, with many different middle segments nonhomologous to those of the first-class and variable-sized inverted repeats that contain only an outer domain. The middle and insertion segments of both classes carry sequences that also are found unassociated from the inverted repeats at many other genomic locations. We conclude that the TU elements are modular structures composed of inverted repeats plus other sequence domains that are themselves members of different families of dispersed repetitive sequences. Such modular elements may have a role in the dispersion and rearrangement of genomic DNA segments. Images PMID:2987685

  6. 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

  7. 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

  8. 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

  9. 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

  10. 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

  11. 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.

  12. Intestinal, portal, and peripheral profiles of daikenchuto (TU-100)'s active ingredients after oral administration

    PubMed Central

    Watanabe, Junko; Kaifuchi, Noriko; Kushida, Hirotaka; Matsumoto, Takashi; Fukutake, Miwako; Nishiyama, Mitsue; Yamamoto, Masahiro; Kono, Toru

    2015-01-01

    A pharmaceutical grade Japanese traditional medicine, daikenchuto (TU-100), consisting of Japanese pepper, processed ginger, and ginseng, has been widely used for various intestinal disorders in Japan and now under development as a new therapeutic drug in the US. It is suggested that TU-100 ingredients exert pharmacological effects on intestines via two routes, from the luminal side before absorption and the peripheral blood stream after absorption. Therefore, in order to fully understand the pharmacological actions of TU-100, it is critically important to know the intraluminal amounts and forms of ingested TU-100 ingredients. In the present study, after administrating TU-100 to rats, the concentrations of TU-100 ingredients and their conjugates in the peripheral and portal blood and ileal contents were determined by LC-MS/MS. Next, TU-100 was administered to patients with ileostomy bags, but whose small intestines are diagnosed as healthy, and the ingredients/conjugates in the ileal effluent were analyzed. The results suggest that: (1) Pepper ingredients hydroxysanshools are rapidly absorbed and enter systemic circulation, (2) Ginseng ingredients ginsenosides are transported to the colon with the least absorption, (3) Ginger ingredients gingerols are absorbed and some conjugated in the small intestine and transported via the portal vein. While only a small amount of gingerols/gingerol conjugates enter systemic circulation, considerable amounts reappear in the small intestine. Thus, the effect of TU-100 on the intestines is believed to be a composite of multiple actions by multiple compounds supplied via multiple routes. PMID:26516578

  13. 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…

  14. 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 ...

  15. Intestinal, portal, and peripheral profiles of daikenchuto (TU-100)'s active ingredients after oral administration.

    PubMed

    Watanabe, Junko; Kaifuchi, Noriko; Kushida, Hirotaka; Matsumoto, Takashi; Fukutake, Miwako; Nishiyama, Mitsue; Yamamoto, Masahiro; Kono, Toru

    2015-10-01

    A pharmaceutical grade Japanese traditional medicine, daikenchuto (TU-100), consisting of Japanese pepper, processed ginger, and ginseng, has been widely used for various intestinal disorders in Japan and now under development as a new therapeutic drug in the US. It is suggested that TU-100 ingredients exert pharmacological effects on intestines via two routes, from the luminal side before absorption and the peripheral blood stream after absorption. Therefore, in order to fully understand the pharmacological actions of TU-100, it is critically important to know the intraluminal amounts and forms of ingested TU-100 ingredients. In the present study, after administrating TU-100 to rats, the concentrations of TU-100 ingredients and their conjugates in the peripheral and portal blood and ileal contents were determined by LC-MS/MS. Next, TU-100 was administered to patients with ileostomy bags, but whose small intestines are diagnosed as healthy, and the ingredients/conjugates in the ileal effluent were analyzed. The results suggest that: (1) Pepper ingredients hydroxysanshools are rapidly absorbed and enter systemic circulation, (2) Ginseng ingredients ginsenosides are transported to the colon with the least absorption, (3) Ginger ingredients gingerols are absorbed and some conjugated in the small intestine and transported via the portal vein. While only a small amount of gingerols/gingerol conjugates enter systemic circulation, considerable amounts reappear in the small intestine. Thus, the effect of TU-100 on the intestines is believed to be a composite of multiple actions by multiple compounds supplied via multiple routes.

  16. The busiest of all ribosomal assistants: elongation factor Tu.

    PubMed

    Kavaliauskas, Darius; Nissen, Poul; Knudsen, Charlotte R

    2012-04-01

    During translation, the nucleic acid language employed by genes is translated into the amino acid language used by proteins. The translator is the ribosome, while the dictionary employed is known as the genetic code. The genetic information is presented to the ribosome in the form of a mRNA, and tRNAs connect the two languages. Translation takes place in three steps: initiation, elongation, and termination. After a protein has been synthesized, the components of the translation apparatus are recycled. During each phase of translation, the ribosome collaborates with specific translation factors, which secure a proper balance between speed and fidelity. Notably, initiation, termination, and ribosomal recycling occur only once per protein produced during normal translation, while the elongation step is repeated a large number of times, corresponding to the number of amino acids constituting the protein of interest. In bacteria, elongation factor Tu plays a central role during the selection of the correct amino acids throughout the elongation phase of translation. Elongation factor Tu is the main subject of this review. PMID:22409271

  17. 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

  18. A constitutive thiamine metabolism mutation, thi80, causing reduced thiamine pyrophosphokinase activity in Saccharomyces cerevisiae.

    PubMed Central

    Nishimura, H; Kawasaki, Y; Nosaka, K; Kaneko, Y; Iwashima, A

    1991-01-01

    We identified a strain carrying a recessive constitutive mutation (thi80-1) with an altered thiamine transport system, thiamine-repressible acid phosphatase, and several enzymes of thiamine synthesis from 2-methyl-4-amino-5-hydroxymethylpyrimidine and 4-methyl-5-beta-hydroxyethylthiazole. The mutant shows markedly reduced activity of thiamine pyrophosphokinase (EC 2.7.6.2) and high resistance to oxythiamine, a thiamine antagonist whose potency depends on thiamine pyrophosphokinase activity. The intracellular thiamine pyrophosphate content of the mutant cells grown with exogenous thiamine (2 x 10(-7) M) was found to be about half that of the wild-type strain under the same conditions. These results suggest that the utilization and synthesis of thiamine in Saccharomyces cerevisiae is controlled negatively by the intracellular thiamine pyrophosphate level. PMID:1849514

  19. 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.

  20. Radioiodination and biodistribution of the monoclonal antibody TU-20 and its scFv fragment

    NASA Astrophysics Data System (ADS)

    Kubaštová, H.; Kleinova, V.; Seifert, D.; Fišer, M.; Kranda, K.

    2006-01-01

    The ability of the monoclonal antibody TU-20 and its scFv fragment to specifically bind to the C-end of the class III beta-tubulin makes these preparations useful as potential diagnostics for in vivo determination of neurodegenerative diseases that entail degradation of neuronal cytoskeleton. To examine this hypothesis, TU-20 and its scFv were labelled with 125I and their properties were extensively investigated. TU-20 and its scFv were labelled via chloramine-T with the yield 90 95% and 64 78%, respectively. Their quality control, performed by an ELISA and gel electrophoresis, determined adequate properties for further studies. The in vitro experiment, involving autoradiography and immunohistochemistry of mice’ brain slices, enabled confirmation of preserved immunospecificity of the radiolabelled substances. Finally, the in vivo biodistribution proved differences in elimination of either TU-20, scFv TU-20, or iodide from the mice.

  1. The human mitochondrial elongation factor tu (EF-Tu) gene: cDNA sequence, genomic localization, genomic structure, and identification of a pseudogene.

    PubMed

    Ling, M; Merante, F; Chen, H S; Duff, C; Duncan, A M; Robinson, B H

    1997-09-15

    The human mitochondrial elongation factor Tu (EF-Tu) is nuclear-encoded and functions in the translational apparatus of mitochondria. The complete human EF-Tu cDNA sequence of 1677 base pairs (bp) with a 101 bp 5'-untranslated region, a 1368 bp coding region, and a 207 bp 3'-untranslated region, has been determined and updated. The predicted protein from this cDNA sequence is approximately 49.8 kDa in size and is composed of 455 amino acids (aa) with a putative N-terminal mitochondrial leader sequence of approximately 50 aa residues. The predicted amino acid sequence shows high similarity to other EF-Tu protein sequences from ox, yeast, and bacteria, and also shows limited similarity to human cystolic elongation factor 1 alpha. The complete size of this cDNA (1677 bp) obtained by cloning and sequencing was confirmed by Northern blot analysis, which showed a single transcript (mRNA) of approximately 1.7 kb in human liver. The genomic structure of this EF-Tu gene has been determined for the first time. This gene contains nine introns with a predicted size of approximately 3.6 kilobases (kb) and has been mapped to chromosome 16p11.2. In addition, an intronless pseudogene of approximately 1.7 kb with 92.6% nucleotide sequence similarity to the EF-Tu gene has also been identified and mapped to chromosome 17q11.2. PMID:9332382

  2. Enacyloxin IIa, an inhibitor of protein biosynthesis that acts on elongation factor Tu and the ribosome.

    PubMed

    Cetin, R; Krab, I M; Anborgh, P H; Cool, R H; Watanabe, T; Sugiyama, T; Izaki, K; Parmeggiani, A

    1996-05-15

    This work analyzes the action of enacyloxin Ila, an inhibitor of bacterial protein biosynthesis. Enacyloxin IIa [IC50 on poly(Phe) synthesis approximately 70 nM] is shown to affect the interaction between elongation factor (EF) Tu and GTP or GDP; in particular, the dissociation of EF-Tu-GTP is strongly retarded, causing the Kd of EF- Tu-GTP to decrease from 500 to 0.7 nM. In its presence, the migration velocity of both GTP- and GDP-bound EF-Tu on native PAGE is increased. The stimulation of EF-Tu-GDP dissociation by EF-Ts is inhibited. EF- Tu-GTP can still form a stable complex with aminoacyl-tRNA (aa-tRNA), but it no longer protects aa-tRNA against spontaneous deacylation, showing that the EF-Tu-GTP orientation with respect to the 3' end of aa-tRNA is modified. However, the EF-Tu-dependent binding of aa-tRNA to the ribosomal A-site is impaired only slightly by the antibiotic and the activity of the peptidyl-transferase center, as determined by puromycin reactivity, is not affected. In contrast, the C-terminal incorporation of Phe into poly(Phe)-tRNA bound to the P-site is inhibited, an effect that is observed if Phe-tRNA is bound to the A-site nonenzymatically as well. Thus, enacyloxin IIa can affect both EF-Tu and the ribosomal A-site directly, inducing an anomalous positioning of aa-tRNA, that inhibits the incorporation of the amino acid into the polypeptide chain. Therefore, it is the first antibiotic found to have a dual specificity targeted to EF-Tu and the ribosome.

  3. 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.

  4. 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.

    2016-10-01

    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.

  5. 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.

  6. Genetic analyses of the host-pathogen system Turnip yellows virus (TuYV)-rapeseed (Brassica napus L.) and development of molecular markers for TuYV-resistance.

    PubMed

    Juergens, Monique; Paetsch, Claudia; Krämer, Ilona; Zahn, Marc; Rabenstein, Frank; Schondelmaier, Jörg; Schliephake, Edgar; Snowdon, Rod; Friedt, Wolfgang; Ordon, Frank

    2010-02-01

    The aphid transmitted Turnip yellows virus (TuYV) has become a serious pathogen in many rapeseed (Brassica napus L.) growing areas. Three-years' field trials were carried out to get detailed information on the genetics of TuYV resistance derived from the resynthesised B. napus line 'R54' and to develop closely linked markers. F(1) plants and segregating doubled-haploid (DH) populations derived from crosses to susceptible cultivars were analysed using artificial inoculation with virus-bearing aphids, followed by DAS-ELISA. Assuming a threshold of E (405) = 0.1 in ELISA carried out in December, the results led to the conclusion that pre-winter inhibition of TuYV is inherited in a monogenic dominant manner. However, the virus titre in most resistant lines increased during the growing period, indicating that the resistance is incomplete and that the level of the virus titre is influenced by environmental factors. Bulked-segregant marker analysis for this resistance locus identified two closely linked SSR markers along with six closely linked and three co-segregating AFLP markers. Two AFLP markers were converted into co-dominant STS markers, facilitating efficient marker-based selection for TuYV resistance. Effective markers are particularly valuable with respect to breeding for TuYV resistance, because artificial inoculation procedures using virus-bearing aphids are extremely difficult to integrate into practical rapeseed breeding programs.

  7. T-cell-activating monoclonal antibodies, reacting with both leukocytes and erythrocytes, recognize the guinea pig Thy-1 differentiation antigen: characterization and cloning of guinea pig CD90.

    PubMed

    Schäfer, H; Bartels, T; Hahn, G; Otto, A; Burger, R

    1999-11-01

    A glycophosphatidylinositol (GPI)-linked differentiation antigen expressed on guinea pig T and B lymphocytes was identified by several monoclonal antibodies; it has been shown previously that this membrane protein induced strong polyclonal T cell proliferation upon antibody binding and costimulation by PMA. Purification by immunoadsorption and microsequencing revealed that this T-cell-activating protein is the homologue of Thy-1 or CD90. In contrast to the Thy-1 antigen of most other species, guinea pig Thy-1 has a much higher molecular weight, which is due to a more extensive N-linked glycosylation, bringing the molecular weight of the total antigen up to 36 kDa. Molecular cloning of guinea pig Thy-1 indicated that the deduced molecular weight of the protein backbone is 12,777 after removal of an N-terminal 19-amino-acid leader peptide and cleavage of the 31 amino acids for GPI anchoring the C-terminal end. Sequence comparison showed that guinea pig Thy-1 has an 82% homology to human and a 72% homology to mouse Thy-1 on the amino acid level. Immunohistological staining of cryostat sections revealed intensive staining with the monoclonal antibody H154 on fibroblasts, fibrocytes, Kupffer cells, alveolar macrophages, and mesangial cells. As observed in the human, mouse, and rat, Thy-1 is abundant in the guinea pig brain. Unlike Thy-1 expression in other species, guinea pig Thy-1 is strongly expressed on most resting, nonactivated B cells and, to a lesser extent, on erythrocytes. While treatment of erythrocytes and lymphocytes with GPI-specific phospholipase C largely decreased reactivity with mAb H154, T cells retained the proliferative response to antibody and phorbol esters.

  8. Marginal Zone B Cells in Neonatal Rats Express Intermediate Levels of CD90 (Thy-1)

    PubMed Central

    Dammers, Peter M.; Lodewijk, Monique E.; Zandvoort, André; Kroese, Frans G. M.

    2002-01-01

    Here we show that marginal zone (MZ)-B cells in rats can already be detected in neonatal spleen from two days after birth. At this time point, morphologically distinct MZs are not present yet and the vast majority of B cells in spleen are located in a concentric area surrounding the T cell zones (PALS). Before MZs are obviously detectable in spleen (14 days after birth), MZ-B cells seem to be enriched at the outer zones of the concentric B cell areas. Similar to adult rats, neonatal MZ-B cells are intermediate-sized cells that express high levels of surface (s)IgM and HIS57 antigen, and low levels of sIgD and CD45R (HIS24). We show here, however, that in contrast to adult MZ-B cells, MZ-B cells (and also recirculating follicular (RF)-B cells) in neonatal rats express higher levels of CD90 (Thy-1). In adult rats, expression of CD90 on the B cell lineage is confined to immature B cells. We speculate that the expression of CD90 on neonatal MZ-B cells may have implications for their responsiveness to polysaccharide (T cell-independent type 2) antigens. PMID:15144015

  9. 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

  10. 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

  11. 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.

  12. 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

  13. Direct evidence of an elongation factor-Tu/Ts·GTP·Aminoacyl-tRNA quaternary complex.

    PubMed

    Burnett, Benjamin J; Altman, Roger B; Ferguson, Angelica; Wasserman, Michael R; Zhou, Zhou; Blanchard, Scott C

    2014-08-22

    During protein synthesis, elongation factor-Tu (EF-Tu) bound to GTP chaperones the entry of aminoacyl-tRNA (aa-tRNA) into actively translating ribosomes. In so doing, EF-Tu increases the rate and fidelity of the translation mechanism. Recent evidence suggests that EF-Ts, the guanosine nucleotide exchange factor for EF-Tu, directly accelerates both the formation and dissociation of the EF-Tu-GTP-Phe-tRNA(Phe) ternary complex (Burnett, B. J., Altman, R. B., Ferrao, R., Alejo, J. L., Kaur, N., Kanji, J., and Blanchard, S. C. (2013) J. Biol. Chem. 288, 13917-13928). A central feature of this model is the existence of a quaternary complex of EF-Tu/Ts·GTP·aa-tRNA(aa). Here, through comparative investigations of phenylalanyl, methionyl, and arginyl ternary complexes, and the development of a strategy to monitor their formation and decay using fluorescence resonance energy transfer, we reveal the generality of this newly described EF-Ts function and the first direct evidence of the transient quaternary complex species. These findings suggest that EF-Ts may regulate ternary complex abundance in the cell through mechanisms that are distinct from its guanosine nucleotide exchange factor functions.

  14. 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.

  15. 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.

  16. Potential bases for some linear values of cooperative TU games

    SciTech Connect

    Dragan, I.

    1994-12-31

    For the Shapley value, S. Hart and A. Mas-Colell (1988, 1989), have introduced the potentials of the value and extended it to the weighted Shapley value. In a paper by Dragan, Potters and Tijs (1989) a basis of the space of TU games with a fixed set of players has been found, such that the coordinates of each game are the potentials of the game and its subgame. In the present paper, we show first that the same approach used for the Shapley value works also for the weighted Shapley value. Then, we use the potential functions introduced recently by Dragan and Driessen (1994) for some fair division rules: the Center of the imputation set (CIS-value), the Egalitarian non-separable contribution (ENSC-value) and the Egalitarian non-average contribution (ENAC-value). We show that the approach used for finding {open_quotes}potential bases{close_quotes} for weighted Shapley values works also for these egalitarian division rules. In each case, we found a potential basis, the values of the basic vectors have been computed and based upon these values a basis for the null space has been determined.

  17. 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

  18. TuBaFrost: European virtual tumor tissue banking.

    PubMed

    Riegman, P H J; Oomen, M H A; Dinjens, W N M; Oosterhuis, J W; Lam, K H; 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; López-Guerrero, J A; Llombart-Bosch, A; Carbone, A; Gloghini, A; Teodorovic, I; Isabelle, M; Passioukov, A; Lejeune, S; Therasse, P; Van Veen, E B

    2006-01-01

    TuBaFrost is a consortium responsible for the task to create a virtual European human frozen tumor tissue bank, composed of high quality frozen tumor tissue collections with corresponding accurate diagnosis stored in European cancer centers and universities, searchable on the Internet, providing rules for access and use and a code of conduct to comply with the various legal and ethical regulations in European countries. Such infrastructure would enlarge tissue availability and accessibility in large amounts of specified or even rare tumor samples. Design of an infrastructure for European residual tissue banking with the described characteristics, clear focus points emerge that can be broken down in dedicated subjects: (1) standardization and quality assurance (QA) to avoid inter-institute quality variation; (2) law and ethics enabling exchange of tissue samples possible between institutes in the different European countries, where law and ethics are characterized by a strong variability; (3) rules for access, with sufficient incentives for collectors; (4) central database application containing innovations on search and selection procedures; (5) support when needed with histology images; and (6) Internet access to search and upload, with in addition a solid website giving proper information on the procedures, intentions and activities not only to the scientific community, but also to the general public. One consortium decision, part of the incentives for collectors, had major impact on the infrastructure; custodianship over the tissues as well as the tissues stay with the collector institute. Resulting in specimens that are not given to an organization, taking decisions on participation of requests, but instead the local collected tissues stay very easy to access by the collector and allows autonomous negotiation between collector and requestor on cooperation, coauthorship in publication or compensation in costs. Thereby, improving availability of large amounts of

  19. Determination of Atmospheric Pressure Loading at TU Vienna

    NASA Astrophysics Data System (ADS)

    Schindelegger, M.; Schuh, H.; Boehm, J.; Wijaya, D.; Karbon, M.

    2010-12-01

    Loading of the Earth's crust due to variations of global atmospheric pressure can displace the positions of geodetic sites by more than 1 cm, both vertically and horizontally on annual to sub-diurnal time scales. Such atmosphere pressure loading (APL) effects have been observed in high-precision space geodetic data, i.e. Global Navigation Satellite Systems (GNSS), Very Long Baseline Interferometry (VLBI), and Satellite Laser Ranging (SLR). These observational data are primarily used for geodynamic studies and it is important to remove the displacement signals due to APL which - if uncorrected - can propagate into other parameters and effects. In order to accurately model APL corrections, we established the project GGOS Atmosphere at TU Vienna. The overall goal of the project is not only to determine APL corrections but also to develop consistent and homogeneous models for atmospheric angular momentum, troposphere delays of radio signals, and gravity field coefficients for the atmosphere. Within project GGOS Atmosphere, we set up an accurate and rigorous procedure for the calculation of APL effects including the unambiguous definition and accurate calculation of a reference pressure. We use operational analysis as well as re-analysis data sets from the European Centre for Medium-Range Weather Forecasts (ECMWF) with a horizontal resolution of 1 degree. The full APL displacements are divided into tidal and non-tidal components and 6-hourly radial and horizontal corrections are provided for all VLBI sites as well as for the nodes of a global 1 degree grid. The displacements are determined in the Center of Mass (CM) and in the Center of the solid Earth (CE) frames. We validate the correction values by VLBI observations and by comparison with results from other providers. Users can freely access the correction values at http://ggosatm.hg.tuwien.ac.at/.

  20. Identification and characterization of Tu88, an antigenic gene from Theileria uilenbergi.

    PubMed

    Lu, Yizhu; Wang, Yuefeng; Li, Youquan; Gou, Huitian; Luo, Jianxun; Yin, Hong; Liu, Zhijie

    2015-06-01

    Theileria uilenbergi is a pathogen that causes ovine theileriosis. Prevention and control of theileriosis relies on its diagnosis at early stages of occurrence and requires understanding of proteins with antigenic properties from the pathogen. Despite its prevalence in China, only a few molecules with antigenic properties have been characterized from T. uilenbergi. In this study, we identified a cDNA named Tu88 by immunoscreening a T. uilenbergi merozoite cDNA library with T. uilenbergi-positive sera from infected sheep. Recombinant Tu88 (rTu88) expressed in bacteria reacted strongly with the positive sera of T. uilenbergi in western blot analysis indicating its potential as an antigen. Southern blot analysis showed that it is a single copy gene. Protein localization by immunostaining blood smears from an infected sheep demonstrated the presence of native Tu88 in merozoites. These findings suggest that Tu88 is a potential candidate antigen for the development of a sero-diagnostic tool.

  1. Preclinical trials in autosomal dominant AD: implementation of the DIAN-TU trial.

    PubMed

    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

    2013-10-01

    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.

  2. 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

  3. 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

  4. 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

  5. [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

  6. Cross-linking of Thy-1 glycoproteins or high-affinity IgE receptors induces mast cell activation via different mechanisms.

    PubMed Central

    Dráberová, L; Dráber, P

    1993-01-01

    Rat peritoneal and pleural mast cells and rat basophilic leukemia cells, RBL-2H3, have been previously shown to be activated by Thy-1-specific monoclonal antibodies (mAb). In the present study we investigated the mechanism of Thy-1-mediated activation and compared it with activation induced by cross-linking of the high-affinity IgE receptor. Binding of an IgG Thy-1 x 1-specific mAb, MRCOX7 (OX7), to RBL-2H3 cells and mast cells, and activation of RBL-2H3 by the OX7 were abrogated by pretreatment of the cells with phosphatidyl inositol-specific phospholipase C (PI-PLC). The F(ab')2 fragment of OX7, in contrast to the Fab' fragment, induced cell activation as well as intact OX7 mAb. Cells sensitized with IgE exhibited an increased responsiveness to anti-Thy-1 antibodies suggesting formation of functional complexes of IgE receptor/IgE/Thy-1/anti-Thy-1. Pretreatment of RBL-2H3 cells with cholera toxin potentiated activation induced by IgE+antigen (Ag) and IgE+OX7, but had no effect on activation induced by OX7 antibody alone. Similarly, dexamethasone had no effect on OX7-induced activation but inhibited IgE+Ag- and IgE+OX7-induced activation. Analysis of phosphotyrosine-containing proteins in RBL-2H3 cell lysates revealed that IgE+Ag and IgE+OX7 induced a marked increase in tyrosine phosphorylation of several proteins that were not tyrosine phosphorylated in cells exposed to OX7 mAb alone. Similar results were obtained when RBL-2H3-derived cells, expressing transfected mouse Thy-1.2, were activated with Thy-1.2-specific IgM antibody. The combined data suggest that Thy-1-specific antibodies activate cells by a mechanism that is different from activation induced by cross-linking of high-affinity IgE receptor. Images Figure 5 Figure 6 PMID:7902332

  7. Effects of Thy-1+ cell depletion on the capacity of donor lymphoid cells to induce tolerance across an entire MHC disparity in sublethally irradiated adult hosts

    SciTech Connect

    Pierce, G.E.; Watts, L.M. )

    1989-08-01

    Thy-1+ cell depletion with anti-Thy-1.2 mAb and complement markedly reduced the capacity of C57BL/6J, H-2b bone marrow to establish mixed lymphoid chimerism and induce tolerance to C57BL/6J skin grafts across an entire MHC disparity in BALB/c, H-2d hosts conditioned with sublethal, fractionated 7.5 Gy total-body irradiation. In this model tolerance can be transferred to secondary irradiated BALB/c hosts only by cells of C57BL/6J donor, not host, genotype isolated from the spleens of tolerant hosts. Thy-1+ cell depletion abolished the capacity of C57BL/6J donor cells from tolerant BALB/c host spleens to transfer tolerance. The capacity of semiallogeneic BALB/c x C57BL/6J F1, H-2d/b donor BM and spleen cells to induce chimerism and tolerance to C57BL/6J skin grafts in BALB/c parental hosts was also reduced by Thy-1+ cell depletion. Thus the requirement for donor Thy-1+ cells cannot be explained simply on the basis of alloaggression. It is unlikely that the requisite Thy-1+ cells are nonspecific suppressor cells: Thy-1+ cell depletion had no effect on the slight but significant prolongation of third-party C3H/HeJ, H-2k skin grafts in irradiated BALB/c hosts injected with allogeneic C57BL/6J or semiallogeneic BALB/c x C57BL/6J F1 BM compared to irradiated controls injected with medium only. Furthermore, injections of semiallogeneic F1 spleen cells had no significant effect on the survival of the third-party grafts, although these cells were fully capable of inducing tolerance, and their capacity to induce tolerance was significantly reduced by Thy-1+ cell depletion. The requirement for a specific population of lymphoid cells, i.e. Thy-1+, remains unexplained but suggests that donor cells might play a role in the induction or maintenance of tolerance in this model other than merely providing a circulating source of donor antigens.

  8. 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...

  9. 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

  10. 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

  11. 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...

  12. 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...

  13. Torsade de pointes and T-U wave alternans associated with arsenic poisoning.

    PubMed

    Little, R E; Kay, G N; Cavender, J B; Epstein, A E; Plumb, V J

    1990-02-01

    Arsenic intoxication is a common form of heavy metal poisoning. Although arsenic-induced circulatory collapse, seizures, and syncope are well known, the potential for serious ventricular arrhythmias is less well recognized. Reported in this study are two cases of arsenic poisoning causing torsade de pointes. Furthermore, marked prolongation of the QT-U interval and the rarely observed phenomenon of T-U wave alternans are demonstrated. Thus, arsenic intoxication may be complicated by prolongation of the QT-U interval and torsade de pointes. T-U wave alternans occurs in the presence of a long QT-U interval and may be an electrocardiographic warning sign of torsade de pointes.

  14. DETECTORS AND EXPERIMENTAL METHODS: Study on the TU gas for the GEM-TPC detector

    NASA Astrophysics Data System (ADS)

    Qi, Hui-Rong; Li, Yu-Lan; Li, Jin; Gao, Yuan-Ning; Li, Yuan-Jing

    2009-04-01

    In this paper several different working gas mixtures for GEM-TPC were evaluated based on a Garfield simulation. Among them, Ar:CH4:CF4 = 90:7:3 (named herein TU gas) was selected for a detailed study because of its better performance. Some performances of drift velocity, transverse diffusion, spatial resolution and the effective number of electrons in various electric fields were obtained. The performance of a GEM-TPC prototype working in the TU gas was studied and compared with that in Ar:CH4 = 90:10 (P10 gas).

  15. 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

  16. Colors of Potentially Hazardous Asteroids 2005 WJ56 and 2007 TU24

    NASA Astrophysics Data System (ADS)

    Betzler, A. S.; Novaes, A. B.; Villa Nova, T. J.; Santos, D.

    2008-09-01

    The near-Earth asteroids 2005 WJ56 and 2007 TU24 were observed by the authors in early 2008 in Salvador, Bahia, Brazil to determine their color indices. For 2005 WJ56, we determined B-V = 0.81 ± 0.09, V-R = 0.48 ± 0.09 and R-I = 0.56 ± 0.09. For 2007 TU24, the V-R color index was estimated to be 0.41 ± 0.02.

  17. 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.

  18. Substrate-induced conformational changes in the S-component ThiT from an energy coupling factor transporter.

    PubMed

    Majsnerowska, Maria; Hänelt, Inga; Wunnicke, Dorith; Schäfer, Lars V; Steinhoff, Heinz-Jürgen; Slotboom, Dirk Jan

    2013-05-01

    Energy coupling factor (ECF) transporters are a recently discovered class of ABC transporters that mediate vitamin uptake in prokaryotes. Characteristic for ECF-type ABC transporters are small integral membrane proteins (S-components) that bind the transported substrates with high affinity. S-components associate with a second membrane protein (EcfT) and two peripheral ATPases to form a complete ATP-dependent transporter. Here, we have used EPR spectroscopy, stopped-flow fluorescence spectroscopy, and molecular dynamics simulations to determine the structural rearrangements that take place in the S-component ThiT from Lactococcus lactis upon binding of thiamin. Thiamin-induced conformational changes were confined to the long and partially membrane-embedded loop between transmembrane helices 1 and 2 that acts as a lid to occlude the binding site. The results indicate that solitary ThiT functions as a bona fide high-affinity substrate binding protein, which lacks a translocation pathway within the protein.

  19. 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

  20. 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

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

    PubMed

    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

    2014-09-16

    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.

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

    PubMed

    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

    2014-09-16

    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

  4. α1-Tubulin FaTuA1 plays crucial roles in vegetative growth and conidiation in Fusarium asiaticum.

    PubMed

    Hu, Weiqun; Zhang, Xiaoping; Chen, Xiang; Zheng, Jingwu; Yin, Yanni; Ma, Zhonghua

    2015-04-01

    The filamentous ascomycete Fusarium asiaticum contains two homologous genes FaTUA1 and FaTUA2 encoding α-tubulins. In this study, we found that FaTUA2 was dispensable for vegetative growth and sporulation in F. asiaticum. The deletion of FaTUA1 however led to dramatically reduced mycelial growth, twisted hyphae and abnormal nuclei in apical cells of hyphae. The FaTUA1 deletion mutant (ΔFaTuA1-5) also showed a significant decrease in conidiation, and produced abnormal conidia. Pathogenicity assays showed that ΔFaTuA1-5 exhibited decreased virulence on wheat head. Unexpectedly, the deletion of FaTUA1 led to resistance to high temperatures. In addition, ΔFaTuA2 showed increased sensitivity to carbendazim. Furthermore, increased FaTUA2 expression in ΔFaTuA1-5 partially restored the defects of the mutant in mycelial growth, conidial production and virulence, vice versa, increased FaTUA1 expression in the FaTUA2 deletion mutant also partially relieved the defect of the mutant in the delay of conidial germination. Taken together, these results indicate that FaTuA1 plays crucial roles in vegetative growth and development, and the functions of FaTuA1 and FaTuA2 are partially interchangeable in F. asiaticum.

  5. How many hair follicles are innervated by one afferent axon? A confocal microscopic analysis of palisade endings in the auricular skin of thy1-YFP transgenic mouse.

    PubMed

    Suzuki, Maasa; Ebara, Satomi; Koike, Taro; Tonomura, Sotatsu; Kumamoto, Kenzo

    2012-01-01

    Hairs are known as a sensory apparatus for touch. Their follicles are innervated predominantly by palisade endings composed of longitudinal and circumferential lanceolate endings. However, little is known as to how their original primary neurons make up a part of the ending. In this study, innervation of the palisade endings was investigated in the auricular skin of thy1-YFP transgenic mouse. Major observations were 1) Only a small portion of PGP9.5-immunopositive axons showed YFP-positivity, 2) All of thy1-YFP-positive sensory axons were thick and myelinated, 3) Individual thy1-YFP-positive trunk axons innervated 4-54 hair follicles, 4) Most palisade endings had a gap of lanceolate ending arrangement, 5) PGP9.5-immunopositive 10-32 longitudinal lanceolate endings were closely arranged. Only a part of them were thy1-YFP-positive axons that originated from 1-3 afferents, and 6) Single nerve bundles of the dermal nerve network included both bidirectional afferents. Palisade endings innervated by multiple sensory neurons might be highly sensitive to hair movement.

  6. Inactivation of thyA in Staphylococcus aureus Attenuates Virulence and Has a Strong Impact on Metabolism and Virulence Gene Expression

    PubMed Central

    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

    2014-01-01

    ABSTRACT 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. PMID:25073642

  7. Identification and purification of human erythroid progenitor cells by monoclonal antibody to the transferrin receptor (TU 67).

    PubMed

    Herrmann, F; Griffin, J D; Sabbath, K D; Oster, W; Wernet, P; Mertelsmann, R

    1988-04-01

    Anti-TU 67 is a murine monoclonal antibody that recognizes the transferrin receptor. With respect to hematopoietic cells TU 67 is expressed by human multipotent colony-forming cells (CFU-Mix), erythroid progenitor cells (BFU-E and CFU-E) and a fraction of granulocyte/monocyte colony forming cells, but is not expressed by mature hematopoietic cells including erythrocytes, platelets, lymphocytes, and peripheral blood myeloid cells. The TU 67-positive fraction of normal bone marrow, separated by fluorescence-activated cell sorting (FACS) or immune rosettes, contained 87% of the erythroid progenitor cells. Erythroid progenitor cells were enriched up to 50-fold by using a combination of monoclonal antibodies to deplete mature hematopoietic cells, followed by positive selection of BFU-E and CFU-E by TU 67 antibody.

  8. 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

  9. 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.

  10. Arabidopsis EF-Tu receptor enhances bacterial disease resistance in transgenic wheat.

    PubMed

    Schoonbeek, Henk-Jan; Wang, Hsi-Hua; Stefanato, Francesca L; Craze, Melanie; Bowden, Sarah; Wallington, Emma; Zipfel, Cyril; Ridout, Christopher J

    2015-04-01

    Perception of pathogen (or microbe)-associated molecular patterns (PAMPs/MAMPs) by pattern recognition receptors (PRRs) is a key component of plant innate immunity. The Arabidopsis PRR EF-Tu receptor (EFR) recognizes the bacterial PAMP elongation factor Tu (EF-Tu) and its derived peptide elf18. Previous work revealed that transgenic expression of AtEFR in Solanaceae confers elf18 responsiveness and broad-spectrum bacterial disease resistance. In this study, we developed a set of bioassays to study the activation of PAMP-triggered immunity (PTI) in wheat. We generated transgenic wheat (Triticum aestivum) plants expressing AtEFR driven by the constitutive rice actin promoter and tested their response to elf18. We show that transgenic expression of AtEFR in wheat confers recognition of elf18, as measured by the induction of immune marker genes and callose deposition. When challenged with the cereal bacterial pathogen Pseudomonas syringae pv. oryzae, transgenic EFR wheat lines had reduced lesion size and bacterial multiplication. These results demonstrate that AtEFR can be transferred successfully from dicot to monocot species, further revealing that immune signalling pathways are conserved across these distant phyla. As novel PRRs are identified, their transfer between plant families represents a useful strategy for enhancing resistance to pathogens in crops.

  11. 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.

  12. 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

  13. 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

  14. 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

  15. Mapping Anatomy to Behavior in Thy1:18 ChR2-YFP Transgenic Mice Using Optogenetics.

    PubMed

    Fenno, Lief E; Gunaydin, Lisa A; Deisseroth, Karl

    2015-06-01

    Linking the activity of defined neural populations with behavior is a key goal of neuroscience. In the context of controlling behavior, electrical stimulation affords researchers precision in the temporal domain with gross regional specificity, whereas pharmacology allows for more specific manipulation of cell types, but in the absence of temporal precision. The use of microbial opsins--light activated, genetically encoded ion channels and pumps--to control mammalian neurons now allows researchers to "sensitize" genetically and/or topologically defined populations of neurons to light to induce either depolarization or hyperpolarization in both a cell-type-specific and temporally precise manner not achievable with previous techniques. Here, we describe the use of transgenic mice expressing the blue-light gated cation channel Channelrhodopsin-2 (ChR2) under control of the Thy1 promoter for the purpose of linking neuronal activity to behavior through restricted delivery of light to an anatomic region of interest. The surgical procedure for implanting a fiber-optic light delivery guide into the mouse brain, the process of optically stimulating the brain in a behaving animal, and post hoc evaluation are given, along with necessary reagents and discussion of common technical problems and their solutions.

  16. 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

  17. 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

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

    2014-06-01

    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

  18. 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

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

    2014-06-01

    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.

  19. Antibacterial activity of thionin Thi2.1 from Arabidopsis thaliana expressed by bovine endothelial cells against Staphylococcus aureus isolates from bovine mastitis.

    PubMed

    Ochoa-Zarzosa, Alejandra; Loeza-Angeles, Heber; Sagrero-Cisneros, Eduardo; Villagómez-Gómez, Erik; Lara-Zárate, Leticia; López-Meza, Joel E

    2008-03-18

    Bovine mastitis is mainly caused by Staphylococcus aureus and antimicrobial therapy, commonly used for its control, has resulted in an increase in the frequency of resistant staphylococci in recent years. Thus, alternative therapies are desirable and the antimicrobial peptides represent attractive control agents. In this work, we expressed the antimicrobial peptide thionin Thi2.1 cDNA from Arabidopsis thaliana in the bovine endothelial cell line BVE-E6E7 and evaluated its activity against bovine mastitis S. aureus isolates. A polyclonal population from BVE-E6E7 cells transfected with the pThi2.1 construct was obtained and thionin Thi2.1 expression was confirmed by RT-PCR. From this population, eight stably transfected cell clones were obtained and their conditioned media (CM) were evaluated against the S. aureus ATCC 27543 strain. Clones showed high antibacterial activity (>95%) relative to the activity of the polyclonal population. The C8 clone showed the highest antibacterial activity (>99%) and its CM was evaluated against eleven bovine mastitis S. aureus isolates. A 2.5microg aliquot of total protein from the C8 clone's CM inhibited the growth of S. aureus isolates (>40%) relative to the CM from BVE-E6E7 cells used as control. Growth inhibition of S. aureus isolates was dose-dependent, showing a total inhibition at concentrations higher than 3.12microg/ml. These results suggest that thionin Thi2.1 antimicrobial peptide could be use in the treatment of bovine mastitis. PMID:17961938

  20. Mapping of fluorescent protein-expressing neurons and axon pathways in adult and developing Thy1-eYFP-H transgenic mice.

    PubMed

    Porrero, Cesar; Rubio-Garrido, Pablo; Avendaño, Carlos; Clascá, Francisco

    2010-07-23

    Transgenic mouse lines in which a fluorescent protein is constitutively expressed under the Thy1 gene promoter have become important models in cell biology and pathology studies of specific neuronal populations. As a result of positional insertion and/or copy number effects on the transgene, the populations expressing the fluorescent protein (eYFP+) vary markedly among the different mice lines. However, identification of the eYFP+ subpopulations has remained sketchy and fragmentary even for the most widely used lines such as Thy1-eYFP-H mice (Feng, G., Mellor, R.H., Bernstein, M., Keller-Peck, C., Nguyen, Q.T., Wallace, M., Nerbonne, J.M., Lichtman and J.W., Sanes. J.R. 2000. Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP. Neuron 28, 41-51). Here, we provide a comprehensive mapping of labeled cell types throughout the central nervous system in adult and postnatal (P0-P30) Thy1-eYFP-H mice. Cell type identification was based on somatodendritic morphology, axon trajectories, and, for cortical cells, retrograde labeling with Fast Blue to distinguish between subpopulations with different axonal targets. In the neocortex, eYFP+ cells are layers 5 and 6 pyramidal neurons, whose abundance and sublaminar distribution varies markedly between areas. Labeling is particularly prevalent in the corticospinal cells; as a result, the pyramidal pathway axons are conspicuously labeled down to the spinal cord. Large populations of hippocampal, subicular and amygdaloid projection neurons are eYFP+ as well. Additional eYFP+ cell groups are located in specific brainstem nuclei. Present results provide a comprehensive reference dataset for adult and developmental studies using the Thy1-eYFP-H mice strain, and show that this animal model may be particularly suitable for studies on the cell biology of corticospinal neurons.

  1. Etudes infrarouge et NMR d'aryl-2 dihydro-1,2 (4H) thiéno[2,3-c]benzo[e]pyrannones-4

    NASA Astrophysics Data System (ADS)

    Robert, J. F.; Ombetta, J. E.; Xicluna, A.; Panouse, J. J.

    The spectroscopic properties (i.r., NMR) of 2-aryl-1,2-dihydro (4H) thiéno[2,3-c]benzo[e]pyrane-4-ones have been studied. Chemical shifts and coupling constants of dihydrothienylic protons are affected by substitutions on the aryl ring and on homocycle 2', 8 and 9 positions to give a conformational approach of these new compounds. The crystal structure of 2-phenyl dihydrothiénocoumarine agrees the results of the spectroscopic study.

  2. Positive Selection of Anti–Thy-1 Autoreactive B-1 Cells and Natural Serum Autoantibody Production Independent from Bone Marrow B Cell Development

    PubMed Central

    Hayakawa, Kyoko; Asano, Masanao; Shinton, Susan A.; Gui, Ming; Wen, Li-Jun; Dashoff, Joni; Hardy, Richard R.

    2003-01-01

    A natural serum autoantibody specific for the Thy-1 glycoprotein (anti–Thy-1 autoantibody [ATA]) is produced by B-1 cells that are positively selected by self-antigen. Here, using ATAμκ transgenic mice we show that cells with this B cell receptor are negatively selected during bone marrow (BM) development. In a Thy-1 null environment, BM ATA B cells progress to a normal follicular stage in spleen. However, in a self-antigen–positive environment, development is arrested at an immature stage in the spleen, concomitant with induction of CD5. Such cells are tolerant and short-lived, different from B-1. Nonetheless, ATA-positive selection was evident by self-antigen–dependent high serum ATA production, comprising ∼90% of serum immunoglobulin M in ATAμκ mice. Splenectomy did not eliminate ATA production and transfer of tolerant splenic B cells did not induce it. These findings demonstrate that B-1 positive selection, resulting in the production of natural serum ATA, arises independently from the major pathway of BM B cell development and selection. PMID:12515816

  3. 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

  4. Russian Tu-144LL SST Flying Laboratory Landing with Drag Chutes at Zhukovsky Air Development Center

    NASA Technical Reports Server (NTRS)

    1997-01-01

    The modified Tupolev Tu-144 supersonic flying laboratory touches down and deploys a trio of drag chutes following a test flight at the Zhukovsky Air Development Center near Moscow, Russia, in July 1997. 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

  5. Russian Tu-144LL SST Flying Laboratory Landing at Zhukovsky Air Development Center

    NASA Technical Reports Server (NTRS)

    1997-01-01

    The Tupolev Tu-144LL supersonic flying laboratory lifts off from the Zhukovsky Air Development Center near Moscow, Russia, on a 1997 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

  6. Russian Tu-144LL SST Flying Laboratory Takeoff at Zhukovsky Air Development Center

    NASA Technical Reports Server (NTRS)

    1997-01-01

    With its nose drooped and canards extended, the Tupolev Tu-144LL supersonic flying laboratory lifts off from the Zhukovsky Air Development Center near Moscow, Russia on a 1997 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

  7. Russian Tu-144LL SST Flying Laboratory Landing at Zhukovsky Air Development Center

    NASA Technical Reports Server (NTRS)

    1997-01-01

    The Tupolev Tu-144LL supersonic flying laboratory touches down at the Zhukovsky Air Development Center near Moscow, Russia, following a 1997 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

  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. 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

  10. 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

  11. 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

  12. 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.

  13. 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

  14. 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…

  15. Know thy journals

    NASA Astrophysics Data System (ADS)

    Kabala, Z. J.

    How do hydrology and water resources journals compare to each other? This question, as noted by McDonnell [1997], is frequently asked by reappointment, tenure, and promotion committees as well as by researchers and graduate students who wonder where to submit their papers. Although the quality of a journal clearly rests with the articles that it publishes, not vice versa, the quality of one's work is often judged in terms of the quality of the journal in which it is published. Therefore, ignoring the journal rankings may be costly regardless of whether or not we deplore the obsession of our society with rankings. What constitutes then a prudent journal choice? Should it be based on the elusive reputation of a journal, its selectivity, its most recent impact factor (defined in the electronic supplement to this article), or other statistics?

  16. 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.

  17. 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.

  18. TuBaFrost 5: multifunctional central database application for a European tumor bank.

    PubMed

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

    2006-12-01

    Developing a tissue bank database has become more than just logically arranging data in tables combined with a search engine. Current demand for high quality samples and data, and the ever-changing legal and ethical regulations mean that the application must reflect TuBaFrost rules and protocols for the collection, exchange and use of tissue. To ensure continuation and extension of the TuBaFrost European tissue bank, the custodianship of the samples, and hence the decision over whether to issue samples to requestors, remains with the local collecting centre. The database application described in this article has been developed to facilitate this open structure virtual tissue bank model serving a large group. It encompasses many key tasks, without the requirement for personnel, hence minimising operational costs. The Internet-accessible database application enables search, selection and request submission for requestors, whereas collectors can upload and edit their collection. Communication between requestor and involved collectors is started with automatically generated e-mails. PMID:17029787

  19. 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

  20. 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.

  1. Tuberculate fruit gene Tu encodes a C2 H2 zinc finger protein that is required for the warty fruit phenotype in cucumber (Cucumis sativus L.).

    PubMed

    Yang, Xuqin; Zhang, Weiwei; He, Huanle; Nie, Jingtao; Bie, Beibei; Zhao, Junlong; Ren, Guoliang; Li, Yue; Zhang, Dabing; Pan, Junsong; Cai, Run

    2014-06-01

    Cucumber fruits that have tubercules and spines (trichomes) are known to possess a warty (Wty) phenotype. In this study, the tuberculate fruit gene Tu was identified by map-based cloning, and was found to encode a transcription factor (TF) with a single C2 H2 zinc finger domain. Tu was identified in all 38 Wty lines examined, and was completely absent from all 56 non-warty (nWty) lines. Cucumber plants transgenic for Tu (TCP) revealed that Tu was required for the Wty fruit phenotype. Subcellular localization showed that the fusion protein GFP-Tu was localized mainly to the nucleus. Based on analyses of semi-quantitative and quantitative reverse transcription polymerase chain reaction (RT-PCR), and mRNA in situ hybridization, we found that Tu was expressed specifically in fruit spine cells during development of fruit tubercules. Moreover, cytokinin (CTK) content measurements and cytological observations in Wty and nWty fruits revealed that the Wty fruit phenotype correlated with high endogenous CTK concentrations. As a result of further analyses on the transcriptomic profile of the nWty fruit epidermis and TCP fruit warts, expression of CTK-associated genes, and hormone content in nWty fruit epidermis, Wty fruit warts and epidermis, and TCP fruit warts and epidermis, we found that Tu probably promoted CTK biosynthesis in fruit warts. Here we show that Tu could not be expressed in the glabrous and tubercule-free mutant line gl that contained Tu, this result that futher confirmed the epistatic effect of the trichome (spine) gene Gl over Tu. Taken together, these data led us to propose a genetic pathway for the Wty fruit trait that could guide future mechanistic studies.

  2. Upregulation of group IB secreted phospholipase A(2) and its M-type receptor in rat ANTI-THY-1 glomerulonephritis.

    PubMed

    Beck, S; Beck, G; Ostendorf, T; Floege, J; Lambeau, G; Nevalainen, T; Radeke, H H; Gurrieri, S; Haas, U; Thorwart, B; Pfeilschifter, J; Kaszkin, M

    2006-10-01

    Treatment of rat glomerular mesangial cell (GMC) cultures with pancreatic secreted phospholipase A(2) (sPLA(2)-IB) results in an enhanced expression of sPLA(2)-IIA and COX-2, possibly via binding to its specific M-type sPLA(2) receptor. In the current study, we have investigated the expression and regulation of sPLA(2)-IB and its receptor during glomerulonephritis (GN). In vivo we used the well-established rat model of anti-Thy 1.1 GN (anti-Thy 1.1-GN) to study the expression of sPLA(2)-IB and the M-type sPLA(2) receptor by immunohistochemistry. In addition, in vitro we determined the interkeukin (IL)-1beta-regulated mRNA and protein expression in primary rat glomerular mesangial and endothelial cells as well as in rat peripheral blood leukocytes (PBLs). Shortly after induction of anti-Thy 1.1-GN, sPLA(2)-IB expression was markedly upregulated in the kidney at 6-24 h. Within glomeruli, the strongest sPLA(2)-IB protein expression was detected on infiltrated granulocytes and monocytes. However, at the same time, the M-type receptor was also markedly upregulated on resident glomerular cells. In vitro, the most prominent cytokine-stimulated secretion of sPLA(2)-IB was observed in monocytes isolated from rat PBLs. Treating glomerular endothelial cells (GECs) with cytokines elicited only weak sPLA(2)-IB expression, but treatment of these cells with exogenous sPLA(2)-IB resulted in a marked expression of the endogenous sPLA(2)-IB. Mesangial cells did not express sPLA(2)-IB at all. The M-type sPLA(2) receptor protein was markedly upregulated on cytokine-stimulated mesangial and endothelial cells as well as on lymphocytes and granulocytes. During anti-Thy 1.1 rat GN, sPLA(2)-IB and the M-type sPLA(2) receptor are induced as primary downstream genes stimulated by inflammatory cytokines. Subsequently, both sPLA(2)-IB and the M-type sPLA(2) receptor are involved in the autocrine and paracrine amplification of the inflammatory process in different resident and infiltrating

  3. 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

  4. Parallelism to solute transport code MT3DMS and case study in TU. Freiberg

    NASA Astrophysics Data System (ADS)

    Abdelaziz, Ramadan; Leb, Hai Ha

    2014-05-01

    A parallel software for 3-D Multi-Species Transport Model MT3DMS was developed. Open Multiprocessing (OpenMP) was used for communication within the processors. MT3DMS emulated the solute transport by dividing the calculation into flow and transport steps. A new preconditioner, derived from Symmetric Successive Over Relaxation (SSOR), is added into the generalized conjugate gradient solver. A case study in the test field at TU Bergakademie Freiberg was used to produce the results and analyze the code performance. A demonstration test field indicated that the parallel mode for MT3DMS is accessible within a processor and problem size. A low timeframe occurs due to speedups for the field test of the solute transport model.

  5. Bone marrow transplantation across major histocompatibility barriers. V. Protection of mice from lethal graft-vs. -host disease by pretreatment of donor cells with monoclonal anti-Thy-1. 2 coupled to the toxin ricin

    SciTech Connect

    Vallera, D.A.; Youle, R.J.; Neville, D.M. Jr.; Kersey, J.H.

    1982-03-01

    A new method has been devised to eliminate T cells from murine bone marrow grafts across major histocompatibility barriers and thus prevent graft-vs.-host disease (GVHD). The method utilizes a monoclonal antibody directed at the Thy-1.2 antigen but is complement independent. To make anti-Thy-1.2 toxic, the antibody is covalently linked to the toxin ricin. Ricin ordinarily binds, enters, and kills cells through receptors containing galactose. The hybrid protein, anti-Thy-1.2-ricin, can enter and kill cells via the Thy-1.2 receptor. In the presence of lactose the usual entry route for ricin is largely blocked and the hybrid is shown to be a highly selective reagent that is T cell specific in its inhibition of mitogen-stimulated splenocytes. We have used a model of severe and fatal GVHD where BALB/c splenocytes and bone marrow cells are given to irradiated C57BL/6 recipients. Over 90% of these mice die by day 70, exhibiting signs of GVHD. When donor cells are pretreated with 0.5 microgram/ml of anti-Thy-1.2-ricin plus 200 mM lactose before injection, 10 of 11 animals survive through day 70 without signs of GVHD. These studies demonstrate that ricin linked to monoclonal antibodies may have utility related to the prevention of GVHD in human bone marrow transplantation.

  6. 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

  7. 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

  8. 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.

  9. 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

  10. 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.

  11. 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

  12. Evolution of translational elongation factor (EF) sequences: reliability of global phylogenies inferred from EF-1 alpha(Tu) and EF-2(G) proteins.

    PubMed Central

    Creti, R; Ceccarelli, E; Bocchetta, M; Sanangelantoni, A M; Tiboni, O; Palm, P; Cammarano, P

    1994-01-01

    The EF-2 coding genes of the Archaea Pyrococcus woesei and Desulfurococcus mobilis were cloned and sequenced. Global phylogenies were inferred by alternative tree-making methods from available EF-2(G) sequence data and contrasted with phylogenies constructed from the more conserved but shorter EF-1 alpha(Tu) sequences. Both the monophyly (sensu Hennig) of Archaea and their subdivision into the kingdoms Crenarchaeota and Euryarchaeota are consistently inferred by analysis of EF-2(G) sequences, usually at a high bootstrap confidence level. In contrast, EF-1 alpha(Tu) phylogenies tend to be inconsistent with one another and show low bootstrap confidence levels. While evolutionary distance and DNA maximum parsimony analyses of EF-1 alpha(Tu) sequences do show archaeal monophyly, protein parsimony and DNA maximum-likelihood analyses of these data do not. In no case, however, do any of the tree topologies inferred from EF-1 alpha(Tu) sequence analyses receive significant bootstrap support. PMID:8159735

  13. 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

  14. 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

  15. Use of standardized SCID-hu Thy/Liv mouse model for preclinical efficacy testing of anti-human immunodeficiency virus type 1 compounds.

    PubMed Central

    Rabin, L; Hincenbergs, M; Moreno, M B; Warren, S; Linquist, V; Datema, R; Charpiot, B; Seifert, J; Kaneshima, H; McCune, J M

    1996-01-01

    We have developed standardized procedures and practices for infection of SCID-hu Thy/Liv mice with human immunodeficiency virus type 1 for the prophylactic administration of antiviral compounds and for evaluation of the antiviral effect in vivo. Endpoint analyses included quantitation of viral load by intracellular p24 enzyme-linked immunosorbent assay, DNA PCR for the presence of proviral genomes, flow cytometry to measure the representation of CD4+ and CD8+ cells, and cocultivation for the isolation of virus. Efficacy tests in this model are demonstrated with the nucleoside analogs zidovudine and dideoxyinosine and with the nonnucleoside reverse transcriptase inhibitor nevirapine. This small-animal model should be particularly useful in the preclinical prioritization of lead compounds within a common chemical class, in the evaluation of alternative in vivo dosing regimens, and in the determination of appropriate combination therapy in vivo. PMID:8851606

  16. Use of standardized SCID-hu Thy/Liv mouse model for preclinical efficacy testing of anti-human immunodeficiency virus type 1 compounds.

    PubMed

    Rabin, L; Hincenbergs, M; Moreno, M B; Warren, S; Linquist, V; Datema, R; Charpiot, B; Seifert, J; Kaneshima, H; McCune, J M

    1996-03-01

    We have developed standardized procedures and practices for infection of SCID-hu Thy/Liv mice with human immunodeficiency virus type 1 for the prophylactic administration of antiviral compounds and for evaluation of the antiviral effect in vivo. Endpoint analyses included quantitation of viral load by intracellular p24 enzyme-linked immunosorbent assay, DNA PCR for the presence of proviral genomes, flow cytometry to measure the representation of CD4+ and CD8+ cells, and cocultivation for the isolation of virus. Efficacy tests in this model are demonstrated with the nucleoside analogs zidovudine and dideoxyinosine and with the nonnucleoside reverse transcriptase inhibitor nevirapine. This small-animal model should be particularly useful in the preclinical prioritization of lead compounds within a common chemical class, in the evaluation of alternative in vivo dosing regimens, and in the determination of appropriate combination therapy in vivo.

  17. High Level of Agreement between Electronic and Paper Mode of Administration of a Thyroid-Specific Patient-Reported Outcome, ThyPRO

    PubMed Central

    Rasmussen, Sofie Larsen; Rejnmark, Lars; Ebbehøj, Eva; Feldt-Rasmussen, Ulla; Rasmussen, Åse Krogh; Bjorner, Jakob Bue; Watt, Torquil

    2016-01-01

    Introduction and Purpose Use of electronic questionnaires to collect health-related quality-of-life data has evolved as an alternative to paper questionnaires. For the electronic questionnaire to be used interchangeably with the validated paper questionnaire, measurement properties similar to the original must be demonstrated. The aim of the present study was to assess the equivalence between the paper version and the electronic version of the thyroid-related quality-of-life questionnaire ThyPRO. Methods Patients with Graves' hyperthyroidism or autoimmune hypothyroidism in a clinically stable phase were included. The patients were recruited from two endocrine outpatient centers. All patients completed both versions in a randomized test-retest set-up. Scores were compared using intraclass correlation coefficients (ICCs), paired t tests and Bland-Altman plots. Limits of agreement were compared with data from a previous paper-paper test-retest study. Results 104 patients were included. ICCs were generally high for the 13 scales, ranging from 0.76 to 0.95. There was a small but significant difference in the scale score between paper and electronic administration for the Cosmetic complaints scale, but no differences were found for any other scale. Bland-Altman plots showed similar limits of agreement compared to the earlier test-retest study of the paper version of ThyPRO. Conclusion Based on our analyses using ICCs, paired t tests and Bland-Altman plots, we found adequate agreement between the paper and electronic questionnaires. The statistically significant difference in score found in the Cosmetic complaints scale is small and probably clinically insignificant. PMID:27099841

  18. Interpretation and discrimination of marshy wetlands by soil factors in the Kuan-Tu Natural Park, Taiwan.

    PubMed

    Liao, Shao-Wei; Chang, Wen-Lian

    2005-08-01

    This work investigated soil samples collected from Kuan-Tu wetlands, Taiwan. Factor analysis was performed to explain the impact of various soil factors on this marshy wetlands located in suburban Taipei. The results indicated that the latent factors were heavy metals, salinity, and soil organic matter. Canonical discriminant analysis was used to improve an existing vegetation classification scheme by identifying the physical-chemical properties of sediment in Kaun-Tu wetlands, Taiwan. Predictive discriminant analysis was used to examine the ability of the models to predict class membership for unknown soil sample. Multivariate analysis of the spatial patterns of soil quality and vegetation types showed that different properties of soil grew different types of vegetation and absorbed contaminants differently. We can feasibly conserve a suitable habitat for wetland biology by processing these unstable predictor variables. The methodology and results provide useful information concerning the Kuan-Tu wetlands and may be applicable to other wetlands with similar properties that are experiencing similar environmental issues.

  19. Genome-wide haploinsufficiency screen reveals a novel role for γ-TuSC in spindle organization and genome stability

    PubMed Central

    Choy, John S.; O'Toole, Eileen; Schuster, Breanna M.; Crisp, Matthew J.; Karpova, Tatiana S.; McNally, James G.; Winey, Mark; Gardner, Melissa K.; Basrai, Munira A.

    2013-01-01

    How subunit dosage contributes to the assembly and function of multimeric complexes is an important question with implications in understanding biochemical, evolutionary, and disease mechanisms. Toward identifying pathways that are susceptible to decreased gene dosage, we performed a genome-wide screen for haploinsufficient (HI) genes that guard against genome instability in Saccharomyces cerevisiae. This led to the identification of all three genes (SPC97, SPC98, and TUB4) encoding the evolutionarily conserved γ-tubulin small complex (γ-TuSC), which nucleates microtubule assembly. We found that hemizygous γ-TuSC mutants exhibit higher rates of chromosome loss and increases in anaphase spindle length and elongation velocities. Fluorescence microscopy, fluorescence recovery after photobleaching, electron tomography, and model convolution simulation of spc98/+ mutants revealed improper regulation of interpolar (iMT) and kinetochore (kMT) microtubules in anaphase. The underlying cause is likely due to reduced levels of Tub4, as overexpression of TUB4 suppressed the spindle and chromosome segregation defects in spc98/+ mutants. We propose that γ-TuSC is crucial for balanced assembly between iMTs and kMTs for spindle organization and accurate chromosome segregation. Taken together, the results show how gene dosage studies provide critical insights into the assembly and function of multisubunit complexes that may not be revealed by using traditional studies with haploid gene deletion or conditional alleles. PMID:23825022

  20. 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.

  1. 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.

  2. TuBaFrost 4: access rules and incentives for a European tumour bank.

    PubMed

    Lopez-Guerrero, J A; Riegman, P H J; Oosterhuis, J W; Lam, K H; 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; Carbone, A; Gloghini, A; Teodorovic, I; Isabelle, M; Passioukov, A; Lejeune, S; Therasse, P; van Veen, E-B; Dinjens, W N M; Llombart-Bosch, A

    2006-11-01

    When designing infrastructure for a networked virtual tumour bank (samples remain at the collector institutes and sample data are collected in a searchable central database), it is apparent that this can only function properly after developing an adequate set of rules for use and access. These rules must include sufficient incentives for the tissue sample collectors to remain active within the network and maintain sufficient sample levels in the local bank. These requirements resulted in a key TuBaFrost rule, stating that the custodianship of the samples remains under the authority of the local collector. As a consequence, the samples and the decision to issue the samples to a requestor are not transferred to a large organisation but instead remain with the collector, thus allowing autonomous negotiation between collector and requestor, potential co-authorship in publications or compensation for collection and processing costs. Furthermore, it realises a streamlined cost effective network, ensuring tissue visibility and accessibility thereby improving the availability of large amounts of samples of highly specific or rare tumour types as well as providing contact opportunities for collaboration between scientists with cutting edge technology and tissue collectors. With this general purpose in mind, the rules and responsibilities for collectors, requestors and central office were generated. PMID:17027256

  3. Planeando Tu Vida: sex and family life education: fundamentals of development, implementation, and evaluation.

    PubMed

    Pick De Weiss, S; Givaudan, M; Givaudan, S

    1993-01-01

    Misinformation about sexuality, reproduction, and contraception is widespread among Mexican adolescents and existing sex education programs have been limited in both scope and availability. To address this situation, the Instituto Mexicano de Investigacion de Familia y Poblacion (IMIFAP) designed a comprehensive sex education program based on data gathered in a 1986 diagnostic survey of 865 adolescents 12-19 years of age and interviews with 365 pregnant adolescents. As part of this preliminary research, one group of teens was exposed to a traditional sex education course while another participated in a program that used participatory learning techniques and emphasized communication skills, assertiveness training, value clarification, peer support, and decision making processes. The latter, more effective approach served as the basis for design of a course, Planeando Tu Vida. Operational evaluations of this course conducted at completion and four and eight months later indicated significant increases in knowledge about contraception, but no effect on age at first intercourse. On the other hand, adolescent males who took the course before onset of sexual activity were significantly more likely to use contraceptives at first intercourse than those in traditional courses. This finding underscores the importance of early initiation of sex education programs. To date, the curriculum has been used in over 100 public and private schools, reaching more than 30,000 adolescents. IMIFAP has since developed more than 70 additional health education course guides aimed at children from preschool through high school, all of which emphasize a participatory approach to learning.

  4. TuBaFrost 6: virtual microscopy in virtual tumour banking.

    PubMed

    Teodorovic, I; Isabelle, M; Carbone, A; Passioukov, A; Lejeune, S; Jaminé, D; Therasse, P; Gloghini, A; Dinjens, W N M; Lam, K H; 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; van Veen, E-B; Oosterhuis, J W; Riegman, P H J

    2006-12-01

    Many systems have already been designed and successfully used for sharing histology images over large distances, without transfer of the original glass slides. Rapid evolution was seen when digital images could be transferred over the Internet. Nowadays, sophisticated Virtual Microscope systems can be acquired, with the capability to quickly scan large batches of glass slides at high magnification and compress and store the large images on disc, which subsequently can be consulted through the Internet. The images are stored on an image server, which can give simple, easy to transfer pictures to the user specifying a certain magnification on any position in the scan. This offers new opportunities in histology review, overcoming the necessity of the dynamic telepathology systems to have compatible software systems and microscopes and in addition, an adequate connection of sufficient bandwidth. Consulting the images now only requires an Internet connection and a computer with a high quality monitor. A system of complete pathology review supporting bio-repositories is described, based on the implementation of this technique in the European Human Frozen Tumor Tissue Bank (TuBaFrost).

  5. Outwitting EF-Tu and the ribosome: translation with d-amino acids

    PubMed Central

    Achenbach, John; Jahnz, Michael; Bethge, Lucas; Paal, Krisztina; Jung, Maria; Schuster, Maja; Albrecht, Renate; Jarosch, Florian; Nierhaus, Knud H.; Klussmann, Sven

    2015-01-01

    Key components of the translational apparatus, i.e. ribosomes, elongation factor EF-Tu and most aminoacyl-tRNA synthetases, are stereoselective and prevent incorporation of d-amino acids (d-aa) into polypeptides. The rare appearance of d-aa in natural polypeptides arises from post-translational modifications or non-ribosomal synthesis. We introduce an in vitro translation system that enables single incorporation of 17 out of 18 tested d-aa into a polypeptide; incorporation of two or three successive d-aa was also observed in several cases. The system consists of wild-type components and d-aa are introduced via artificially charged, unmodified tRNAGly that was selected according to the rules of ‘thermodynamic compensation’. The results reveal an unexpected plasticity of the ribosomal peptidyltransferase center and thus shed new light on the mechanism of chiral discrimination during translation. Furthermore, ribosomal incorporation of d-aa into polypeptides may greatly expand the armamentarium of in vitro translation towards the identification of peptides and proteins with new properties and functions. PMID:26026160

  6. 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

  7. 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

  8. 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

  9. 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

  10. Tu-144LL SST Flying Laboratory Landing on Runway at Zhukovsky Air Development Center near Moscow, Ru

    NASA Technical Reports Server (NTRS)

    1998-01-01

    The Tupolev Tu-144LL SST Flying Laboratory rolls down the runway at the Zhukovsky Air Development Center near Moscow, Russia, after 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

  11. 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

  12. 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

  13. 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

  14. Molecular cloning and sequence determination of the nuclear gene coding for mitochondrial elongation factor Tu of Saccharomyces cerevisiae.

    PubMed

    Nagata, S; Tsunetsugu-Yokota, Y; Naito, A; Kaziro, Y

    1983-10-01

    A 3.1-kilobase Bgl II fragment of Saccharomyces cerevisiae carrying the nuclear gene encoding the mitochondrial polypeptide chain elongation factor (EF) Tu has been cloned on pBR327 to yield a chimeric plasmid pYYB. The identification of the gene designated as tufM was based on the cross-hybridization with the Escherichia coli tufB gene, under low stringency conditions. The complete nucleotide sequence of the yeast tufM gene was established together with its 5'- and 3'-flanking regions. The sequence contained 1,311 nucleotides coding for a protein of 437 amino acids with a calculated Mr of 47,980. The nucleotide sequence and the deduced amino acid sequence of tufM were 60% and 66% homologous, respectively, to the corresponding sequences of E. coli tufA, when aligned to obtain the maximal homology. Plasmid YRpYB was then constructed by cloning the 2.5-kilobase EcoRI fragment of pYYB carrying tufM into a yeast cloning vector YRp-7. A mRNA hybridizable with tufM was isolated from the total mRNA of S. cerevisiae D13-1A transformed with YRpYB and translated in the reticulocyte lysate. The mRNA could direct the synthesis of a protein with Mr 48,000, which was immunoprecipitated with an anti-E. coli EF-Tu antibody but not with an antibody against yeast cytoplasmic EF-1 alpha. The results indicate that the tufM gene is a nuclear gene coding for the yeast mitochondrial EF-Tu. PMID:6353412

  15. 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

  16. 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

  17. Reducing ppGpp level rescues an extreme growth defect caused by mutant EF-Tu.

    PubMed

    Bergman, Jessica M; Hammarlöf, Disa L; Hughes, Diarmaid

    2014-01-01

    Transcription and translation of mRNA's are coordinated processes in bacteria. We have previously shown that a mutant form of EF-Tu (Gln125Arg) in Salmonella Typhimurium with a reduced affinity for aa-tRNA, causes ribosome pausing, resulting in an increased rate of RNase E-mediated mRNA cleavage, causing extremely slow growth, even on rich medium. The slow growth phenotype is reversed by mutations that reduce RNase E activity. Here we asked whether the slow growth phenotype could be reversed by overexpression of a wild-type gene. We identified spoT (encoding ppGpp synthetase/hydrolase) as a gene that partially reversed the slow growth rate when overexpressed. We found that the slow-growing mutant had an abnormally high basal level of ppGpp that was reduced when spoT was overexpressed. Inactivating relA (encoding the ribosome-associated ppGpp synthetase) also reduced ppGpp levels and significantly increased growth rate. Because RelA responds specifically to deacylated tRNA in the ribosomal A-site this suggested that the tuf mutant had an increased level of deacylated tRNA relative to the wild-type. To test this hypothesis we measured the relative acylation levels of 4 families of tRNAs and found that proline isoacceptors were acylated at a lower level in the mutant strain relative to the wild-type. In addition, the level of the proS tRNA synthetase mRNA was significantly lower in the mutant strain. We suggest that an increased level of deacylated tRNA in the mutant strain stimulates RelA-mediated ppGpp production, causing changes in transcription pattern that are inappropriate for rich media conditions, and contributing to slow growth rate. Reducing ppGpp levels, by altering the activity of either SpoT or RelA, removes one cause of the slow growth and reveals the interconnectedness of intracellular regulatory mechanisms.

  18. 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.

  19. 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.

  20. 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

  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. Crosslinking of elongation factor Tu to tRNA(Phe) by trans-diamminedichloroplatinum (II). Characterization of two crosslinking sites in the tRNA.

    PubMed Central

    Wikman, F P; Romby, P; Metz, M H; Reinbolt, J; Clark, B F; Ebel, J P; Ehresmann, C; Ehresmann, B

    1987-01-01

    Trans-diamminedichloroplatinum (II) was used to induce reversible crosslinks between EF-Tu and Phe-tRNA(Phe) within the ternary EF-Tu/GTP/Phe-tRNA(Phe) complex. Up to 40% of the complex was specifically converted into crosslinked species. Two crosslinking sites have been unambiguously identified. The major one encompassing nucleotides 58 to 65 is located in the 3'-part of the T-stem, and the minor one encompassing nucleotides 31 to 42 includes the anticodon loop and part of the 3'-strand of the anticodon stem. Images PMID:3302946

  4. The TU-025 Keishibukuryogan for Hot Flash Management in Postmenopausal Women: Results and Lessons for Future Research

    PubMed Central

    Plotnikoff, Gregory A.; Watanabe, Kenji; Torkelson, Carolyn; La Valleur, June; Radosevich, David M.

    2011-01-01

    Objective To assess the efficacy of TU-025, keishibukuryogan, a Japanese prescription herbal medicine used for hot flash management, in American women. Methods This randomized, double-blind, placebo-controlled, phase II trial enrolled 178 postmenopausal women, aged 45–58 years, with a Mayo hot flash score greater than 28 per week and who met other inclusion criteria. After a one-week placebo run-in period, participants were randomly assigned to either 12 weeks of placebo, 7.5 g/day or 12.5 g/day. Primary and secondary outcomes were measured by the Mayo Clinic Hot Flash Diary, the Greene Climacteric Index and the Pittsburgh Sleep Quality Index. Results At three months, hot flash scores, climacteric symptoms and sleep quality improved by 34% in the placebo group, 40% in the 7.5 g/day group and 38% in the 12.5 g/day group. (p < 0.001). However, the differences in changes between groups were not statistically significant (p = 0.990). Diarrhea unexpectedly developed in 20% of participants receiving active medication. Conclusions For American women, unlike the clinical experience for Japanese women, TU-025 did not significantly reduce the frequency and severity of hot flash symptoms, improve climacteric symptoms or benefit sleep quality. This study identified several potentially significant methodological factors to be considered in future scientific assessments of traditional Asian medicines. PMID:21738077

  5. 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

  6. 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

  7. 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

  8. 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

  9. 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

  10. 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

  11. 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.

  12. 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

  13. 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

  14. The Ca(2+)/Calmodulin/CaMKK2 Axis: Nature's Metabolic CaMshaft.

    PubMed

    Marcelo, Kathrina L; Means, Anthony R; York, Brian

    2016-10-01

    Calcium (Ca(2+)) is an essential ligand that binds its primary intracellular receptor calmodulin (CaM) to trigger a variety of downstream processes and pathways. Central to the actions of Ca(2+)/CaM is the activation of a highly conserved Ca(2+)/CaM kinase (CaMK) cascade that amplifies Ca(2+) signals through a series of subsequent phosphorylation events. Proper regulation of Ca(2+) flux is necessary for whole-body metabolism and disruption of Ca(2+) homeostasis has been linked to various metabolic diseases. Here we provide a synthesis of recent advances that highlight the roles of the Ca(2+)/CaMK axis in key metabolic tissues. An appreciation of this information is critical to understanding the mechanisms by which Ca(2+)/CaM-dependent signaling contributes to metabolic homeostasis and disease.

  15. :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 ...

  16. Phylogenetic analysis of evolutionary relationships of the planctomycete division of the domain bacteria based on amino acid sequences of elongation factor Tu.

    PubMed

    Jenkins, C; Fuerst, J A

    2001-05-01

    Sequences from the tuf gene coding for the elongation factor EF-Tu were amplified and sequenced from the genomic DNA of Pirellula marina and Isosphaera pallida, two species of bacteria within the order Planctomycetales. A near-complete (1140-bp) sequence was obtained from Pi. marina and a partial (759-bp) sequence was obtained for I. pallida. Alignment of the deduced Pi. marina EF-Tu amino acid sequence against reference sequences demonstrated the presence of a unique 11-amino acid sequence motif not present in any other division of the domain Bacteria. Pi. marina shared the highest percentage amino acid sequence identity with I. pallida but showed only a low percentage identity with other members of the domain Bacteria. This is consistent with the concept of the planctomycetes as a unique division of the Bacteria. Neither primary sequence comparison of EF-Tu nor phylogenetic analysis supports any close relationship between planctomycetes and the chlamydiae, which has previously been postulated on the basis of 16S rRNA. Phylogenetic analysis of aligned EF-Tu amino acid sequences performed using distance, maximum-parsimony, and maximum-likelihood approaches yielded contradictory results with respect to the position of planctomycetes relative to other bacteria. It is hypothesized that long-branch attraction effects due to unequal evolutionary rates and mutational saturation effects may account for some of the contradictions. PMID:11443344

  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. Honor thy gut symbionts redux.

    PubMed

    Gordon, Jeffrey I

    2012-06-01

    Exploring our gut microbial communities with new tools is allowing us to revisit old questions; to develop new concepts about our evolution, postnatal development, systems physiology, individuality, and definitions of health; and to further delineate the impact of our changing life-styles. It is also allowing us to envision exciting new ways for addressing global health problems. This area is inherently interdisciplinary, offering a wealth of opportunities to create new fields, partnerships, and educational initiatives. It is captivating to the public and carries substantial expectations. As such, participating scientists need to sponsor proactive, solution-focused discussions of its societal implications.

  19. 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.

  20. Analysis of transgenic wheat (Triticum aestivum L.) harboring a maize (Zea mays L.) gene for plastid EF-Tu: segregation pattern, expression and effects of the transgene.

    PubMed

    Fu, Jianming; Ristic, Zoran

    2010-06-01

    We previously reported that transgenic wheat (Triticum aestivum L.) carrying a maize (Zea mays L.) gene (Zmeftu1) for chloroplast protein synthesis elongation factor, EF-Tu, displays reduced thermal aggregation of leaf proteins, reduced injury to photosynthetic membranes (thylakoids), and enhanced rate of CO(2) fixation following exposure to heat stress (18 h at 45 degrees C) [Fu et al. in Plant Mol Biol 68:277-288, 2008]. In the current study, we investigated the segregation pattern and expression of the transgene Zmeftu1 and determined the grain yield of transgenic plants after exposure to a brief heat stress (18 h at 45 degrees C). We also assessed thermal aggregation of soluble leaf proteins in transgenic plants, testing the hypothesis that increased levels of EF-Tu will lead to a non-specific protection of leaf proteins against thermal aggregation. The transgenic wheat displayed a single-gene pattern of segregation of Zmeftu1. Zmeftu1 was expressed, and the transgenic plants synthesized and accumulated three anti-EF-Tu cross-reacting polypeptides of similar molecular mass but different pI, suggesting the possibility of posttranslational modification of this protein. The transgenic plants also showed better grain yield after exposure to heat stress compared with their non-transgenic counterparts. Soluble leaf proteins of various molecular masses displayed lower thermal aggregation in transgenic than in non-transgenic wheat. The results suggest that overexpression of chloroplast EF-Tu can be beneficial to wheat tolerance to heat stress. Moreover, the results also support the hypothesis that EF-Tu contributes to heat tolerance by acting as a molecular chaperone and protecting heat-labile proteins from thermal aggregation in a non-specific manner.

  1. [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

  2. Atypical archaeal tRNA pyrrolysine transcript behaves towards EF-Tu as a typical elongator tRNA

    PubMed Central

    Théobald-Dietrich, Anne; Frugier, Magali; Giegé, Richard; Rudinger-Thirion, Joëlle

    2004-01-01

    The newly discovered tRNAPyl is involved in specific incorporation of pyrrolysine in the active site of methylamine methyltransferases in the archaeon Methanosarcina barkeri. In solution probing experiments, a transcript derived from tRNAPyl displays a secondary fold slightly different from the canonical cloverleaf and interestingly similar to that of bovine mitochondrial tRNASer(uga). Aminoacylation of tRNAPyl transcript by a typical class II synthetase, LysRS from yeast, was possible when its amber anticodon CUA was mutated into a lysine UUU anticodon. Hydrolysis protection assays show that lysylated tRNAPyl can be recognized by bacterial elongation factor. This indicates that no antideterminant sequence is present in the body of the tRNAPyl transcript to prevent it from interacting with EF-Tu, in contrast with the otherwise functionally similar tRNASec that mediates selenocysteine incorporation. PMID:14872064

  3. Origin of 48Ca

    NASA Astrophysics Data System (ADS)

    Clayton, Donald D.; Krishnan, Tracy D.; Meyer, Bradley S.

    1997-02-01

    We stress conceptual understanding of why 48Ca survives low-entropy expansions but not high-entropy expansions. Type Ia supernovae are thereby demonstrated to be its source. The interdependence of the quasiequilibrium cluster (QSE) and the number of heavy nuclei is the key. By varying reaction rates, we show that nuclear cross sections are not a sensitive need for the problem of 48Ca nucleosynthesis.

  4. 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

  5. Identification of novel lymphoid tissues in murine intestinal mucosa where clusters of c-kit+ IL-7R+ Thy1+ lympho-hemopoietic progenitors develop

    PubMed Central

    1996-01-01

    We have revealed that about one and a half thousand tiny clusters, filled with one thousand closely packed lymphocytes, can be found throughout the murine small and large intestinal mucosa. They are located in crypt lamina propria (cryptopatches; CP) and can be first detected at 14-17 d after birth. A large fraction of lymphocytes in CP expresses c-kit, IL-7R, Thy1 and a lymphocyte function-associated antigen, LFA-1, whereas most of them remain CD3-, TCR alpha beta-, TCR gamma delta-, sIgM-, and B220-. The population size of IL-2R alpha+, HSA+ and Pgp-1+ subsets is variable (20-50%) and the composition of CD8+, Ly-1+, and CD4+ subsets is smaller but also variable (3-20%). In the small intestine, CP do not contain cells undergoing apoptosis nor cells bearing RAG-1 molecules, but do contain dendritic stromal cells bearing CD11c/CD18 molecules. The frequency of DNA replicating cells in CP is higher than that in Peyer's patches (PP), is lower than that in the thymic cortex and is almost comparable with that in the thymic medulla. The numbers of CP remain the same in aged mice (> 114 wk) but double after estrogen treatment even though the thymi are attenuated sharply in both conditions. Thus, with respect to histogenesis, lymphocyte composition and tissue level of cellular behavior, neither PP, isolated lymphoid follicles, peripheral LNs, nor thymus are identical with CP. Finally, CP are virtually absent in lamina propria of IL-7R-deficient mice that display a profound reduction in thymic and peripheral lymphoid cellularity. By contrast, CP are present in germ- free mice and in athymic (nu/nu), SCID, TCR beta x delta-/-, RAG-2-/-, PP-deficient (aly/aly), stem cell factor (Sl/Sld) and c-kit (W/Wv) mutant mice. Taking all of these results together, CP are the first identification of gut-associated murine lymphoid tissues where the generation of IL-7-dependent lympho-hematopoietic progenitors for T and/or B cell descendants may start to take place at the age of

  6. 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

  7. 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

  8. 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

  9. 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

  10. Heterologous expression of a plastid EF-Tu reduces protein thermal aggregation and enhances CO2 fixation in wheat (Triticum aestivum) following heat stress.

    PubMed

    Fu, Jianming; Momcilović, Ivana; Clemente, Thomas E; Nersesian, Natalya; Trick, Harold N; Ristic, Zoran

    2008-10-01

    Heat stress is a major constraint to wheat production and negatively impacts grain quality, causing tremendous economic losses, and may become a more troublesome factor due to global warming. At the cellular level, heat stress causes denaturation and aggregation of proteins and injury to membranes leading to alterations in metabolic fluxes. Protein aggregation is irreversible, and protection of proteins from thermal aggregation is a strategy a cell uses to tolerate heat stress. Here we report on the development of transgenic wheat (Triticum aestivum) events, expressing a maize gene coding for plastidal protein synthesis elongation factor (EF-Tu), which, compared to non-transgenic plants, display reduced thermal aggregation of leaf proteins, reduced heat injury to photosynthetic membranes (thylakoids), and enhanced rate of CO(2) fixation after exposure to heat stress. The results support the concept that EF-Tu ameliorates negative effects of heat stress by acting as a molecular chaperone. This is the first demonstration of the introduction of a plastidal EF-Tu in plants that leads to protection against heat injury and enhanced photosynthesis after heat stress. This is also the first demonstration that a gene other than HSP gene can be used for improvement of heat tolerance and that the improvement is possible in a species that has a complex genome, hexaploid wheat. The results strongly suggest that heat tolerance of wheat, and possibly other crop plants, can be improved by modulating expression of plastidal EF-Tu and/or by selection of genotypes with increased endogenous levels of this protein.

  11. 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

  12. 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

  13. 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.

  14. Loss of epidermal integrity by T cell-mediated attack induces long-term local resistance to subsequent attack. I. Induction of resistance correlates with increases in Thy-1+ epidermal cell numbers

    PubMed Central

    1990-01-01

    The cutaneous graft-versus-host disease (GVHD) lesions induced by intradermal injection of cloned autoreactive T cells have been shown to subside rapidly and the epidermis returns to normal 2 wk after injection. Those mice that had spontaneously recovered from the cutaneous GVHD became resistant to subsequent attempts to induce the cutaneous GVHD by the T cells while maintaining their activity to mount delayed-type hypersensitivity (DTH) responses and to induce the enlargement of the popliteal lymph nodes (PLN). The resistance appeared to be restricted to the epidermal structures of the injection sites, suggesting the involvement of locally acting suppression mechanisms. This local resistance was not specific for the clonotype used for the induction of the resistance. A loss of the epidermal integrity by an attack of T cells capable of producing cutaneous GVHD was a prerequisite for the induction of the resistance. By up to at least 8 mo after injection of the T cells, no mice became susceptible to the cutaneous GVHD again, provided that the T cells were injected into the same footpad sites that had initially received the T cells. This resistance correlated well with the great increase (20-30-fold) in Thy- 1+ EC number. The great increase in the number of Thy-1+ EC following destruction of epidermal structures may be important in protecting the epidermal integrity from an additional attack by T cells. PMID:1969918

  15. 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.

  16. 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

  17. Increasing HIV-related knowledge, communication, and testing intentions among Latinos: Protege tu Familia: Hazte la Prueba.

    PubMed

    Rios-Ellis, Britt; Espinoza, Lilia; Bird, Mara; Garcia, Melawhy; D'Anna, Laura Hoyt; Bellamy, Laura; Scolari, Rosana

    2010-08-01

    Latinos are less likely to be aware of their HIV seropositivity than African Americans and Whites. 'Protege tu Familia: Hazte la Prueba' is a culturally and linguistically-sensitive HIV/AIDS prevention and testing program targeting Latino families. Using community-based participatory research techniques, Spanish-speaking bicultural community health workers helped develop and then used an educational flip chart and materials to conduct outreach and HIV prevention education in diverse settings. The intervention was created to increase HIV/AIDS-related knowledge, to improve communication regarding sexual risk, and to augment intentions to use condoms and test for HIV. A secondary purpose was to decrease HIV-related stigma by improving knowledge about transmission and reducing homophobia. Participants demonstrated significant increases in HIV knowledge, intention to practice safer sex and communicate sexual risk to partner(s), and intention to test for HIV. Improvements were also found in self-reported comfort levels when interacting with and caring for the HIV positive, thus decreasing HIV/AIDS-related stigma.

  18. Los Angeles, CA, USA

    NASA Technical Reports Server (NTRS)

    1991-01-01

    This rare cloud and smog free view of Los Angeles, CA (34.0N, 118.5W) is a result of strong Santa Ana winds blowing from the east. Both cultural and natural features are well displayed and all of the major streets, highways and freeways can be traced in their entirety throughout the city as well as the major business and commercial sections. On the eastern edge of the scene, the San Andreas fault cuts across from southeast to northwest.

  19. Sulfide capacities of CaO-CaF2-CaCl2 melts

    NASA Astrophysics Data System (ADS)

    Simeonov, Simeon; Sakai, Toshihiko; Maeda, Masafumi

    1992-06-01

    The sulfide capacityC_{s^{2 - } } = ({text{pct S}}^{{text{2 - }}} )(p_{{text{O}}_{text{2}} } /p_{{text{S}}_{text{2}} } )^{1/2} ) of CaO-CaF2-CaCl2 slag was determined at temperatures from 1000 °C to 1300 °C by equilibrating molten slag, molten silver, and CO-CO2-Ar gas mixture. The sulfide capacity increases with replacing CaCl2 by CaF2 in slags of constant CaO contents. The sulfide capacity also increases with increasing temperature as well as with increasing CaO content at a constant ratio of CaF2/CaCl2 of unity. A linear relationship between the sulfide capacity and carbonate capacity in literature was observed on a logarithmic scale.

  20. Measurement of CA1P and CA in leaves

    SciTech Connect

    Moore, B.d.; Kobza, J.; Seemann, J.R. )

    1990-05-01

    Carboxyarabinitol-1-phosphate (CA1P) and carboxyarabinitol (CA) were assayed in leaves by isotope dilution. {sup 14}C-labeled standards were synthesized from (2-{sup 14}C) CABP using acid (CA1P) or alkaline (CA) phosphatase. Either was added to boiling 80% EtOH along with liquid N{sub 2}-killed leaves. Each was largely purified by anion exchange chromatography. CA1P samples were subjected to 2D-TLE/TLC. The specific activity of the {sup 14}C-containing spot was measured using alkaline phosphatase. CA samples were run on an HPLC and the specific activity was determined using a UV monitor and a flow-through radioisotope detector. In 3 of the tested species, light/dark amount of CA1P (nmol/mg Chl) were kidney bean, 0.7/67; sugar beet, 0.8/33; and Alocasia, 0/3.4. Light/dark CA levels (nmol/mg Chl) in these respective species were 897/653, 3.2/3.5, and 5.7/4.6. These results support the hypothesis that CA is a product of CA1P metabolism in vivo under high light, but also indicate that CA is not the only intermediate involved in CA1P synthesis under low light/dark conditions.

  1. 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

  2. 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.

  3. Ca(2+) Binding and Transport Studied with Ca(2+)/EGTA Buffers and (45)Ca(2+).

    PubMed

    Sehgal, Pankaj; Olesen, Claus; Møller, Jesper V

    2016-01-01

    The chapter describes procedures useful for determination of Ca(2+) binding by membranous Ca(2+)-ATPase based on the correction for the removal of Ca(2+) present in a non-bound state in the suspension medium. This is done by a filtration procedure that retains the membranous material on the Millipore filters. With suitable sucking devices it is possible to gently remove without dehydration nearly all medium from the Ca(2+) containing membranes, except that required for wetting of the filters on which they are deposited. Correction for this effect can be done with a double-filter where the radioactive content of the lower (protein-free) filter is subtracted from that present in the upper filter for calculation of Ca(2+) binding. This methodology can be used to study the effect of inhibitors on Ca(2+) binding and -transport, and with Ca(2+)/EGTA buffers to explore the Ca(2+) binding affinities and cooperative aspects of the two transport sites.

  4. 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

  5. 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

  6. 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

  7. 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

  8. 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.

  9. 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

  10. Elongation Factor Tu Prevents Misediting of Gly-tRNA(Gly) Caused by the Design Behind the Chiral Proofreading Site of D-Aminoacyl-tRNA Deacylase

    PubMed Central

    Routh, Satya Brata; Ahmad, Sadeem; Suma, Katta; Kumar, Mantu; Kuncha, Santosh Kumar; Yadav, Kranthikumar; Kruparani, Shobha P; Sankaranarayanan, Rajan

    2016-01-01

    D-aminoacyl-tRNA deacylase (DTD) removes D-amino acids mischarged on tRNAs and is thus implicated in enforcing homochirality in proteins. Previously, we proposed that selective capture of D-aminoacyl-tRNA by DTD’s invariant, cross-subunit Gly-cisPro motif forms the mechanistic basis for its enantioselectivity. We now show, using nuclear magnetic resonance (NMR) spectroscopy-based binding studies followed by biochemical assays with both bacterial and eukaryotic systems, that DTD effectively misedits Gly-tRNAGly. High-resolution crystal structure reveals that the architecture of DTD’s chiral proofreading site is completely porous to achiral glycine. Hence, L-chiral rejection is the only design principle on which DTD functions, unlike other chiral-specific enzymes such as D-amino acid oxidases, which are specific for D-enantiomers. Competition assays with elongation factor thermo unstable (EF-Tu) and DTD demonstrate that EF-Tu precludes Gly-tRNAGly misediting at normal cellular concentrations. However, even slightly higher DTD levels overcome this protection conferred by EF-Tu, thus resulting in significant depletion of Gly-tRNAGly. Our in vitro observations are substantiated by cell-based studies in Escherichia coli that show that overexpression of DTD causes cellular toxicity, which is largely rescued upon glycine supplementation. Furthermore, we provide direct evidence that DTD is an RNA-based catalyst, since it uses only the terminal 2′-OH of tRNA for catalysis without the involvement of protein side chains. The study therefore provides a unique paradigm of enzyme action for substrate selection/specificity by DTD, and thus explains the underlying cause of DTD’s activity on Gly-tRNAGly. It also gives a molecular and functional basis for the necessity and the observed tight regulation of DTD levels, thereby preventing cellular toxicity due to misediting. PMID:27224426

  11. [Organizational problems of disaster victim identification in mass casualties as exemplified by Tu 154-M and Airbus A310 passenger plane crashes].

    PubMed

    Volkov, A V; Kolkutin, V V; Klevno, V A; Shkol'nikov, B V; Kornienko, I V

    2008-01-01

    Managerial experience is described that was gained during the large-scale work on victim identification following mass casualties in the Tu 154-M and Airbus A310 passenger plane crashes. The authors emphasize the necessity to set up a specialized agency of constant readiness meeting modern requirements for the implementation of a system of measures for personality identification. This agency must incorporate relevant departments of the Ministries of Health, Defense, and Emergency Situations as well as investigative authorities and other organizations. PMID:19048869

  12. Expression level and immunolocalization of de novo methyltransferase 3 protein (TuDNMT3) in adult females and males of the two-spotted spider mite, Tetranychus urticae.

    PubMed

    Yang, Si-Xia; Guo, Chao; Zhang, Yan-Kai; Sun, Jing-Tao; Hong, Xiao-Yue

    2015-11-01

    DNA methylation is an epigenetic mechanism for regulating developmental and other important processes in eukaryotes. Several essential components of the DNA methylation machinery have been identified, such as DNA methyltransferases. In the two-spotted spider mite, Tetranychus urticae Koch, we have identified one DNA methyltransferase 3 gene (Tudnmt3) and tentatively investigated its potential role in adult females and males. Here, to better elucidate the functional role of Tudnmt3, its protein structure, expression and localization were subjected to more detailed analyses. Bioinformatic analyses clearly showed that the structure of TuDNMT3 was highly conserved, with several vital amino acid residues for the activation and stabilization of its confirmation. Western blot analyses revealed that this protein was expressed in both genders, with higher expression in adult females, which was inconsistent with the gene expression, suggesting translational regulation of Tudnmt3. Subsequent immunodetection provided supportive evidence for higher expression of the TuDNMT3 protein in adult females and indicated that this protein was generally localized in the cytoplasm and that its expression was predominantly confined to the genital region of spider mites, strengthening the hypothesis that de novo methylation mediated by Tudnmt3 in gonad development or gametogenesis has a different mechanism from maintenance methyltransferase.

  13. GTPase domains of ras p21 oncogene protein and elongation factor Tu: analysis of three-dimensional structures, sequence families, and functional sites.

    PubMed

    Valencia, A; Kjeldgaard, M; Pai, E F; Sander, C

    1991-06-15

    GTPase domains are functional and structural units employed as molecular switches in a variety of important cellular functions, such as growth control, protein biosynthesis, and membrane traffic. Amino acid sequences of more than 100 members of different subfamilies are known, but crystal structures of only mammalian ras p21 and bacterial elongation factor Tu have been determined. After optimal superposition of these remarkably similar structures, careful multiple sequence alignment, and calculation of residue-residue interactions, we analyzed the two subfamilies in terms of structural conservation, sequence conservation, and residue contact strength. There are three main results. (i) A structure-based alignment of p21 and elongation factor Tu. (ii) The definition of a common conserved structural core that may be useful as the basis of model building by homology of the three-dimensional structure of any GTPase domain. (iii) Identification of sequence regions, other than the effector loop and the nucleotide binding site, that may be involved in the functional cycle: they are loop L4, known to change conformation after GTP hydrolysis; helix alpha 2, especially Arg-73 and Met-67 in ras p21; loops L8 and L10, including ras p21 Arg-123, Lys-147, and Leu-120; and residues located spatially near the N and C termini. These regions are candidate sites for interaction either with the GTP/GDP exchange factor, with a GTPase-affected function, or with a molecule delivered to a destination site with the aid of the GTPase domain.

  14. 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

  15. Electromagnetic modelling, inversion and data-processing techniques for GPR: ongoing activities in Working Group 3 of COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Pajewski, Lara; Giannopoulos, Antonis; van der Kruk, Jan

    2015-04-01

    This work aims at presenting the ongoing research activities carried out in Working Group 3 (WG3) 'EM methods for near-field scattering problems by buried structures; data processing techniques' of the COST (European COoperation in Science and Technology) Action TU1208 'Civil Engineering Applications of Ground Penetrating Radar' (www.GPRadar.eu). The principal goal of the COST Action TU1208 is to exchange and increase scientific-technical knowledge and experience of GPR techniques in civil engineering, simultaneously promoting throughout Europe the effective use of this safe and non-destructive technique in the monitoring of infrastructures and structures. WG3 is structured in four Projects. Project 3.1 deals with 'Electromagnetic modelling for GPR applications.' Project 3.2 is concerned with 'Inversion and imaging techniques for GPR applications.' The topic of Project 3.3 is the 'Development of intrinsic models for describing near-field antenna effects, including antenna-medium coupling, for improved radar data processing using full-wave inversion.' Project 3.4 focuses on 'Advanced GPR data-processing algorithms.' Electromagnetic modeling tools that are being developed and improved include the Finite-Difference Time-Domain (FDTD) technique and the spectral domain Cylindrical-Wave Approach (CWA). One of the well-known freeware and versatile FDTD simulators is GprMax that enables an improved realistic representation of the soil/material hosting the sought structures and of the GPR antennas. Here, input/output tools are being developed to ease the definition of scenarios and the visualisation of numerical results. The CWA expresses the field scattered by subsurface two-dimensional targets with arbitrary cross-section as a sum of cylindrical waves. In this way, the interaction is taken into account of multiple scattered fields within the medium hosting the sought targets. Recently, the method has been extended to deal with through-the-wall scenarios. One of the

  16. TU-F-18C-08: Micro-Calcification Detectability Using Spectral Breast CT Based On a Si Strip Detector

    SciTech Connect

    Cho, H; Ding, H; Molloi, S; Barber, W; Iwanczyk, J

    2014-06-15

    Purpose: To investigate the feasibility of micro-calcification (μCa) detectability by using an energy-resolved photon-counting Si strip detector for spectral breast computed tomography (CT). Methods: A bench-top CT system was constructed using a tungsten anode x-ray source with a focal spot size of 0.8 mm and a single line 256-pixel Si strip photon counting detector with a pixel pitch of 100 μm. The slice thickness was 0.5 mm. Five different size groups of calcium carbonate grains, from 105 to 215 μm in diameter, were embedded in a cylindrical resin phantom with a diameter of 16 mm to simulate μCas. The phantoms were imaged at 65 kVp with an Entrance Skin Air Kerma (ESAK) of 1.2, 3, 6, and 8 mGy. The images were reconstructed using a standard filtered back projection (FBP) with a ramp filter. A total of 200 μCa images (5 different sizes of μCas × 4 different doses × 10 images for each setting) were combined with another 200 control images without μCas, to ultimately form 400 images for the reader study. The images were displayed in random order to three blinded observers, who were asked to give a binary score on each image regarding the presence of μCas. The μCa detectability for each image was evaluated in terms of binary decision theory metrics. The sensitivity, specificity, and accuracy were calculated to study the size and dose-dependence for μCa detectability. Additionally, the influence of the partial volume effect on the μCa detectability was investigated by simulation. Results: For a μCa larger than 140 μm in diameter, detection accuracy of above 90 % was achieved with the investigated prototype spectral CT system at ESAK of 1.2 mGy. Conclusion: The proposed Si strip detector is expected to offer superior image quality with the capability to detect μCas for low dose breast imaging.

  17. CaPTC Biennial Meetings

    Cancer.gov

    CaPTC hosts the 'Biennial Science of Global Prostate Cancer Disparities in Black Men' conference to address the growing global public health problem of prostate cancer among Black men in industrialized and developing countries.

  18. CaMKII-dependent SR Ca leak contributes to doxorubicin-induced impaired Ca handling in isolated cardiac myocytes

    PubMed Central

    Sag, Can M.; Köhler, Anne C.; Anderson, Mark E.; Backs, Johannes; Maier, Lars S.

    2011-01-01

    Objective Doxorubicin (DOX) is one of the most effective chemotherapeutic agents, but cardiotoxicity limits DOX therapy. Although the mechanisms are not entirely understood, reactive oxygen species (ROS) appear to be involved in DOX cardiotoxicity. Ca/calmodulin dependent protein kinase II (CaMKII) can be activated by ROS through oxidation and is known to contribute to myocardial dysfunction through Ca leakage from the sarcoplasmic reticulum (SR). Rationale We hypothesized that CaMKII contributes to DOX-induced defects in intracellular Ca ([Ca]i) handling. Methods Cardiac myocytes were isolated from wild-type (WT) adult rat hearts and from mouse hearts lacking the predominant myocardial CaMKII isoform (CaMKIIδ−/−, KO) vs. WT. Isolated cardiomyocytes were investigated 30 min after DOX (10 µmol/L) superfusion, using epifluorescence and confocal microscopy. Intracellular ROS-generation ([ROS]i) and [Ca]i handling properties were assessed. In a subset of experiments, KN-93 or AIP (each 1 µmol/L) were used to inhibit CaMKII. Melatonin (Mel, 100 µmol/L) served as ROS-scavenger. Western blots were performed to determine the amount of CaMKII phosphorylation and oxidation. Results DOX increased [ROS]i and led to significant diastolic [Ca]i overload in rat myocytes. This was associated with reduced [Ca]i transients, a 5.8-fold increased diastolic SR Ca leak and diminished SR Ca content. ROS-scavenging partially rescued Ca handling. Western blots revealed increased CaMKII phosphorylation, but not CaMKII oxidation after DOX. Pharmacological CaMKII inhibition attenuated diastolic [Ca]i overload after DOX superfusion and led to partially restored [Ca]i transients and SR Ca content, presumably due to reduced Ca spark frequency. In line with this concept, isoform-specific CaMKIIδ-KO attenuated diastolic [Ca]i overload and Ca spark frequency. Conclusions DOX exposure induces CaMKII-dependent SR Ca leakage, which partially contributes to impaired cellular [Ca]i homeostasis

  19. [CA 125--a tumor marker?].

    PubMed

    Pabst, T; Ludwig, C

    1995-06-17

    Tumor markers are useful tools in monitoring malignancies postoperatively or under hormone-/chemotherapy. In contrast, they usually lack diagnostic relevance and uncritical use may result in confusing situations. We describe three cases of diagnostic determinations of the tumor marker CA 125 resulting in subsequent partially invasive procedures. Based on these three cases, serum CA 125 levels were examined in 49 patients with abdominal diseases. We found CA 125 to be less a tumor product than an unspecific expression of stimulated mesothelial cells of the peritoneum. CA 125 was a marker for ascites (16 of 16 patients) and an indicator of infra-diaphragmatic involvement in non-Hodgkin's lymphoma (11 of 12 patients). Furthermore, 5 of 6 patients with inflammatory abdominal diseases showed elevated CA 125 levels, as did 13 of 15 patients with solid abdominal tumors of different histology (all non-ovarian cancer, no ascites). In conclusion, CA 125 remains a good marker for follow-up of ovarian cancer, but should not be used for diagnosis of abdominal processes.

  20. COST Action TU1206 "SUB-URBAN - A European network to improve understanding and use of the ground beneath our cities"

    NASA Astrophysics Data System (ADS)

    Campbell, Diarmad; de Beer, Johannes; Lawrence, David; van der Meulen, Michiel; Mielby, Susie; Hay, David; Scanlon, Ray; Campenhout, Ignace; Taugs, Renate; Eriksson, Ingelov

    2014-05-01

    Sustainable urbanisation is the focus of SUB-URBAN, a European Cooperation in Science and Technology (COST) Action TU1206 - A European network to improve understanding and use of the ground beneath our cities. This aims to transform relationships between experts who develop urban subsurface geoscience knowledge - principally national Geological Survey Organisations (GSOs), and those who can most benefit from it - urban decision makers, planners, practitioners and the wider research community. Under COST's Transport and Urban Development Domain, SUB-URBAN has established a network of GSOs and other researchers in over 20 countries, to draw together and evaluate collective urban geoscience research in 3D/4D characterisation, prediction and visualisation. Knowledge exchange between researchers and City-partners within 'SUB-URBAN' is already facilitating new city-scale subsurface projects, and is developing a tool-box of good-practice guidance, decision-support tools, and cost-effective methodologies that are appropriate to local needs and circumstances. These are intended to act as catalysts in the transformation of relationships between geoscientists and urban decision-makers more generally. As a result, the importance of the urban sub-surface in the sustainable development of our cities will be better appreciated, and the conflicting demands currently placed on it will be acknowledged, and resolved appropriately. Existing city-scale 3D/4D model exemplars are being developed by partners in the UK (Glasgow, London), Germany (Hamburg) and France (Paris). These draw on extensive ground investigation (10s-100s of thousands of boreholes) and other data. Model linkage enables prediction of groundwater, heat, SuDS, and engineering properties. Combined subsurface and above-ground (CityGML, BIMs) models are in preparation. These models will provide valuable tools for more holistic urban planning; identifying subsurface opportunities and saving costs by reducing uncertainty in

  1. Rejection of tmRNA·SmpB after GTP hydrolysis by EF-Tu on ribosomes stalled on intact mRNA.

    PubMed

    Kurita, Daisuke; Miller, Mickey R; Muto, Akira; Buskirk, Allen R; Himeno, Hyouta

    2014-11-01

    Messenger RNAs lacking a stop codon trap ribosomes at their 3' ends, depleting the pool of ribosomes available for protein synthesis. In bacteria, a remarkable quality control system rescues and recycles stalled ribosomes in a process known as trans-translation. Acting as a tRNA, transfer-messenger RNA (tmRNA) is aminoacylated, delivered by EF-Tu to the ribosomal A site, and accepts the nascent polypeptide. Translation then resumes on a reading frame within tmRNA, encoding a short peptide tag that targets the nascent peptide for degradation by proteases. One unsolved issue in trans-translation is how tmRNA and its protein partner SmpB preferentially recognize stalled ribosomes and not actively translating ones. Here, we examine the effect of the length of the 3' extension of mRNA on each step of trans-translation by pre-steady-state kinetic methods and fluorescence polarization binding assays. Unexpectedly, EF-Tu activation and GTP hydrolysis occur rapidly regardless of the length of the mRNA, although the peptidyl transfer to tmRNA decreases as the mRNA 3' extension increases and the tmRNA·SmpB binds less tightly to the ribosome with an mRNA having a long 3' extension. From these results, we conclude that the tmRNA·SmpB complex dissociates during accommodation due to competition between the downstream mRNA and the C-terminal tail for the mRNA channel. Rejection of the tmRNA·SmpB complex during accommodation is reminiscent of the rejection of near-cognate tRNA from the ribosome in canonical translation.

  2. GPR surveying of transport infrastructures and buildings; underground utility and void sensing - ongoing activities in Working Group 2 of COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Pajewski, Lara; Plati, Christina; Derobert, Xavier

    2015-04-01

    This work aims at presenting the ongoing research activities carried out in Working Group 2 'GPR surveying of pavements, bridges, tunnels and buildings; underground utility and void sensing' of the COST (European COoperation in Science and Technology) Action TU1208 'Civil Engineering Applications of Ground Penetrating Radar' (www.GPRadar.eu). The principal goal of the COST Action TU1208 is to exchange and increase scientific-technical knowledge and experience of Ground Penetrating Radar (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. Four Working Groups (WGs) carry out the research activities. WG1 focuses on the development of innovative GPR equipment dedicated for civil engineering applications. WG2 deals with the development of guidelines and protocols for the surveying, through the use of a GPR system, of transport infrastructure and buildings, as well as for the sensing of utilities and voids. WG3 deals with the development of electromagnetic forward and inverse scattering methods, for the characterization of GPR scenarios, as well as with data- processing algorithms for the elaboration of the data collected during GPR surveys. WG4 is concerned with the use of GPR in fields different from the civil engineering, as well as with the integration of GPR with other non-destructive testing techniques. Each WG includes several Projects. WG2 includes five Projects. Project 2.1 focuses on outlining 'Innovative inspection procedures for effective GPR surveying of critical transport infrastructures (pavements, bridges and tunnels).' Project 2.2 is concerned with the development of 'Innovative inspection procedures for effective GPR surveying of buildings.' Project 2.3 deals with identifying 'Innovative inspection procedures for effective GPR sensing and mapping of underground utilities and voids, with a focus to urban

  3. Ca2+ scraps: local depletions of free [Ca2+] in cardiac sarcoplasmic reticulum during contractions leave substantial Ca2+ reserve.

    PubMed

    Shannon, Thomas R; Guo, Tao; Bers, Donald M

    2003-07-11

    Free [Ca2+] inside the sarcoplasmic reticulum ([Ca2+]SR) is difficult to measure yet critically important in controlling many cellular systems. In cardiac myocytes, [Ca2+]SR regulates cardiac contractility. We directly measure [Ca2+]SR in intact cardiac myocytes dynamically and quantitatively during beats, with high spatial resolution. Diastolic [Ca2+]SR (1 to 1.5 mmol/L) is only partially depleted (24% to 63%) during contraction. There is little temporal delay in the decline in [Ca2+]SR at release junctions and between junctions, indicating rapid internal diffusion. The incomplete local Ca2+ release shows that the inherently positive feedback of Ca2+-induced Ca2+ release terminates, despite a large residual driving force. These findings place stringent novel constraints on how excitation-contraction coupling works in heart and also reveal a Ca2+ store reserve that could in principle be a therapeutic target to enhance cardiac function in heart failure.

  4. The site of net absorption of Ca from the intestinal tract of growing pigs and effect of phytic acid, Ca level and Ca source on Ca digestibility.

    PubMed

    González-Vega, J Caroline; Walk, Carrie L; Liu, Yanhong; Stein, Hans H

    2014-01-01

    An experiment was conducted to test the hypothesis that the standardised digestibility of Ca in calcium carbonate and Lithothamnium calcareum Ca is not different regardless of the level of dietary Ca, and that phytic acid affects the digestibility of Ca in these two ingredients to the same degree. The objectives were to determine where in the intestinal tract Ca absorption takes place and if there are measurable quantities of basal endogenous Ca fluxes in the stomach, small intestine or large intestine. Diets contained calcium carbonate or L. calcareum Ca as the sole source of Ca, 0% or 1% phytic acid and 0.4% or 0.8% Ca. A Ca-free diet was also formulated and used to measure endogenous fluxes and losses of Ca. Nine growing pigs (initial body weight 23.8 ± 1.3 kg) were cannulated in the duodenum and in the distal ileum, and faecal, ileal and duodenal samples were collected. Duodenal endogenous fluxes of Ca were greater (p < 0.05) than ileal endogenous fluxes and total tract endogenous losses of Ca, but ileal endogenous fluxes were less (p < 0.05) than total tract endogenous losses. Standardised digestibility of Ca was not affected by the level of phytic acid, but decreased (p < 0.05) as Ca level increased in L. calcareum Ca diets, but that was not the case if calcium carbonate was the source of Ca (interaction, p < 0.05). The standardised duodenal digestibility (SDD), standardised ileal digestibility (SID) and standardised total tract digestibility (STTD) of Ca were not different if calcium carbonate was the source of dietary Ca. However, the STTD of Ca in L. calcareum Ca was greater (p < 0.05) than the SID and SDD of Ca. The SDD, SID and STTD of Ca in calcium carbonate were greater (p < 0.05) than those of L. calcareum Ca. In conclusion, under the conditions of this experiment, standardised digestibility of Ca is not affected by the level of phytic acid, but may be affected by dietary Ca level depending on the Ca source. Calcium from calcium carbonate is mostly

  5. Routes of Ca2+ Shuttling during Ca2+ Oscillations: FOCUS ON THE ROLE OF MITOCHONDRIAL Ca2+ HANDLING AND CYTOSOLIC Ca2+ BUFFERS.

    PubMed

    Pecze, László; Blum, Walter; Schwaller, Beat

    2015-11-20

    In some cell types, Ca(2+) oscillations are strictly dependent on Ca(2+) influx across the plasma membrane, whereas in others, oscillations also persist in the absence of Ca(2+) influx. We observed that, in primary mesothelial cells, the plasmalemmal Ca(2+) influx played a pivotal role. However, when the Ca(2+) transport across the plasma membrane by the "lanthanum insulation method" was blocked prior to the induction of the serum-induced Ca(2+) oscillations, mitochondrial Ca(2+) transport was found to be able to substitute for the plasmalemmal Ca(2+) exchange function, thus rendering the oscillations independent of extracellular Ca(2+). However, in a physiological situation, the Ca(2+)-buffering capacity of mitochondria was found not to be essential for Ca(2+) oscillations. Moreover, brief spontaneous Ca(2+) changes were observed in the mitochondrial Ca(2+) concentration without apparent changes in the cytosolic Ca(2+) concentration, indicating the presence of a mitochondrial autonomous Ca(2+) signaling mechanism. In the presence of calretinin, a Ca(2+)-buffering protein, the amplitude of cytosolic spikes during oscillations was decreased, and the amount of Ca(2+) 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 Ca(2+) shuttling pathways in primary mesothelial cells during Ca(2+) oscillations: Ca(2+) shuttled between (i) the endoplasmic reticulum (ER) and mitochondria, (ii) the ER and the extracellular space, and (iii) the ER and cytoplasmic Ca(2+) buffers.

  6. Controls on Sr/Ca and Mg/Ca in scleractinian corals: The effects of Ca-ATPase and transcellular Ca channels on skeletal chemistry

    NASA Astrophysics Data System (ADS)

    Emif Allison, Nicola; Cohen, Itay; Finch, Adrian A.; Erez, Jonathan

    2011-11-01

    The Sr/Ca of aragonitic coral skeletons is a commonly used palaeothermometer. However skeletal Sr/Ca is typically dominated by weekly-monthly oscillations which do not reflect temperature or seawater composition and the origins of which are currently unknown. To test the impact of transcellular Ca2+ transport processes on skeletal Sr/Ca, colonies of the branching coral, Pocillopora damicornis, were cultured in the presence of inhibitors of Ca-ATPase (ruthenium red) and Ca channels (verapamil hydrochloride). The photosynthesis, respiration and calcification rates of the colonies were monitored throughout the experiment. The skeleton deposited in the presence of the inhibitors was identified (by 42Ca spike) and analysed for Sr/Ca and Mg/Ca by secondary ion mass spectrometry. The Sr/Ca of the aragonite deposited in the presence of either of the inhibitors was not significantly different from that of the solvent (dimethyl sulfoxide) control, although the coral calcification rate was reduced by up to 66% and 73% in the ruthenium red and verapamil treatments, respectively. The typical precision (95% confidence limits) of mean Sr/Ca determinations within any treatment was <±1% and differences in skeletal Sr/Ca between treatments were correspondingly small. Either Ca-ATPase and Ca channels transport Sr2+ and Ca2+ in virtually the same ratio in which they are present in seawater or transcellular processes contribute little Ca2+ to the skeleton and most Ca is derived from seawater transported directly to the calcification site. Variations in the activities of Ca-ATPase and Ca-channels are not responsible for the weekly-monthly Sr/Ca oscillations observed in skeletal chronologies, assuming that the specificities of Ca transcellular transport processes are similar between coral genera.

  7. Controls on Sr/Ca and Mg/Ca in scleractinian corals: The effects of Ca-ATPase and transcellular Ca channels on skeletal chemistry

    NASA Astrophysics Data System (ADS)

    Allison, Nicola; Cohen, Itay; Finch, Adrian A.; Erez, Jonathan; EMIF

    2011-11-01

    The Sr/Ca of aragonitic coral skeletons is a commonly used palaeothermometer. However skeletal Sr/Ca is typically dominated by weekly-monthly oscillations which do not reflect temperature or seawater composition and the origins of which are currently unknown. To test the impact of transcellular Ca 2+ transport processes on skeletal Sr/Ca, colonies of the branching coral, Pocillopora damicornis, were cultured in the presence of inhibitors of Ca-ATPase (ruthenium red) and Ca channels (verapamil hydrochloride). The photosynthesis, respiration and calcification rates of the colonies were monitored throughout the experiment. The skeleton deposited in the presence of the inhibitors was identified (by 42Ca spike) and analysed for Sr/Ca and Mg/Ca by secondary ion mass spectrometry. The Sr/Ca of the aragonite deposited in the presence of either of the inhibitors was not significantly different from that of the solvent (dimethyl sulfoxide) control, although the coral calcification rate was reduced by up to 66% and 73% in the ruthenium red and verapamil treatments, respectively. The typical precision (95% confidence limits) of mean Sr/Ca determinations within any treatment was <±1% and differences in skeletal Sr/Ca between treatments were correspondingly small. Either Ca-ATPase and Ca channels transport Sr 2+ and Ca 2+ in virtually the same ratio in which they are present in seawater or transcellular processes contribute little Ca 2+ to the skeleton and most Ca is derived from seawater transported directly to the calcification site. Variations in the activities of Ca-ATPase and Ca-channels are not responsible for the weekly-monthly Sr/Ca oscillations observed in skeletal chronologies, assuming that the specificities of Ca transcellular transport processes are similar between coral genera.

  8. Large Ca2+-dependent facilitation of CaV2.1 channels revealed by Ca2+ photo-uncaging

    PubMed Central

    Lee, Shin-Rong; Adams, Paul J; Yue, David T

    2015-01-01

    Key points CaV2.1 channels constitute a dominant Ca2+ entry pathway into brain neurons, triggering downstream Ca2+-dependent processes such as neurotransmitter release. CaV2.1 is itself modulated by Ca2+, resulting in activity-dependent enhancement of channel opening termed Ca2+-dependent facilitation (CDF). Real-time Ca2+ imaging and Ca2+ uncaging here reveal that CDF turns out to be strikingly faster, more Ca2+ sensitive, and larger than anticipated on previous grounds. Robust resolution of the quantitative profile of CDF enables deduction of a realistic biophysical model for this process. These results suggest that CaV2.1 CDF would figure most prominently in short-term synaptic plasticity and cerebellar Purkinje cell rhythmicity. Abstract CaV2.1 (P-type) voltage-gated Ca2+ channels constitute a major source of neuronal Ca2+ current, strongly influencing rhythmicity and triggering neurotransmitter release throughout the central nervous system. Fitting with such stature among Ca2+ entry pathways, CaV2.1 is itself feedback regulated by intracellular Ca2+, acting through calmodulin to facilitate channel opening. The precise neurophysiological role of this calcium-dependent facilitation (CDF) remains uncertain, however, in large measure because the very magnitude, Ca2+ dependence and kinetics of CDF have resisted quantification by conventional means. Here, we utilize the photo-uncaging of Ca2+ with CaV2.1 channels fluxing Li+ currents, so that voltage-dependent activation of channel gating is no longer conflated with Ca2+ entry, and CDF is then driven solely by light-induced increases in Ca2+. By using this strategy, we now find that CDF can be unexpectedly large, enhancing currents by as much as twofold at physiological voltages. CDF is steeply Ca2+ dependent, with a Hill coefficient of approximately two, a half-maximal effect reached by nearly 500 nm Ca2+, and Ca2+ on/off kinetics in the order of milliseconds to tens of milliseconds. These properties were

  9. How Does Stochastic Ryanodine Receptor-Mediated Ca Leak Fail to Initiate a Ca Spark?

    PubMed Central

    Sato, Daisuke; Bers, Donald M.

    2011-01-01

    Spontaneous calcium (Ca) sparks are initiated by single ryanodine receptor (RyR) opening. Once one RyR channel opens, it elevates local [Ca] in the cleft space ([Ca]Cleft), which opens other RyR channels in the same Ca release unit (CaRU) via Ca-induced Ca-release. Experiments by Zima et al. (J. Physiol. 588:4743–4757, 2010) demonstrate that spontaneous Ca sparks occur only when intrasarcoplasmic-reticulum (SR) [Ca] ([Ca]SR) is above a threshold level, but that RyR-mediated SR Ca leak exists without Ca sparks well below this threshold [Ca]SR. We examine here how single RyR opening at lower [Ca]SR can fail to recruit Ca sparks at a CaRU, while still contributing to SR Ca leak. We assess this using a physiologically detailed mathematical model of junctional SR Ca release in which RyR gating is regulated by [Ca]SR and [Ca]Cleft. We find that several factors contribute to the failure of Ca sparks as [Ca]SR declines: 1), lower [Ca]SR reduces driving force and thus limits local [Ca]Cleft achieved and the rate of rise during RyR opening; 2), low [Ca]SR limits RyR open time (τO), which further reduces local [Ca]Cleft attained; 3), low τO and fast [Ca]Cleft dissipation after RyR closure shorten the opportunity for neighboring RyR activation; 4), at low [Ca]SR, the RyR exhibits reduced [Ca]Cleft sensitivity. We conclude that all of these factors conspire to reduce the probability of Ca sparks as [Ca]SR declines, despite continued RyR-mediated Ca leak. In addition, these same factors explain the much lower efficacy of L-type Ca channel opening to trigger local SR Ca release at low [Ca]SR during excitation-contraction coupling. Conversely, all of these factors are fundamentally important for increasing the propensity for pro-arrhythmic Ca sparks and waves in cardiac myocytes at high [Ca]SR. PMID:22098735

  10. Solar Ca II K Observations

    NASA Astrophysics Data System (ADS)

    Bertello, Luca; Pevtsov, Alexei A.; Tlatov, Andrey; Singh, Jagdev

    2016-07-01

    Some of the most important archives of past and current long-term solar synoptic observations in the resonance line of Ca II K are described here. These observations are very important for understanding the state of the solar magnetism on time scales up to several decades. The first observations of this kind began in 1904 at the Kodaikanal Observatory (India), followed by similar programs at different other locations. Regular full-disk Ca II K monitoring programs started in 1915 at the Mount Wilson Observatory (USA) and in 1917 at the National Solar Observatory of Japan. Beginning in 1919 and in 1926 regular observations were taken also at the Paris-Meudon Observatory (France) and at the "Donati solar tower telescope of the Arcetri Astrophysical Observatory in Italy, respectively. In 1926 the the Astronomical Observatory of the Coimbra University in Portugal started its own program of Ca II K observations. Although some of these programs have been terminated over the years, their data archives constitute a unique resource for studies of solar variability. In the early 1970s, the National Solar Observatory (NSO) at Sacramento Peak (USA) started a new program of daily Sun-as-a-star observations in the Ca II K line. Today the NSO is continuing these observations through its Synoptic Optical Long-term Investigations of the Sun (SOLIS) facility.

  11. Design and testing of Ground Penetrating Radar equipment dedicated for civil engineering applications: ongoing activities in Working Group 1 of COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Pajewski, Lara; Manacorda, Guido; Persico, Raffaele

    2015-04-01

    This work aims at presenting the ongoing research activities carried out in Working Group 1 'Novel GPR instrumentation' of the COST (European COoperation in Science and Technology) Action TU1208 'Civil Engineering Applications of Ground Penetrating Radar' (www.GPRadar.eu). The principal goal of the COST Action TU1208 is to exchange and increase scientific-technical knowledge and experience of GPR techniques in civil engineering, simultaneously promoting throughout Europe the effective use of this safe and non-destructive technique in the monitoring of infrastructures and structures. Working Group 1 (WG1) of the Action focuses on the development of innovative GPR equipment dedicated for civil engineering applications. It includes three Projects. Project 1.1 is focused on the 'Design, realisation and optimisation of innovative GPR equipment for the monitoring of critical transport infrastructures and buildings, and for the sensing of underground utilities and voids.' Project 1.2 is concerned with the 'Development and definition of advanced testing, calibration and stability procedures and protocols, for GPR equipment.' Project 1.3 deals with the 'Design, modelling and optimisation of GPR antennas.' During the first year of the Action, WG1 Members coordinated between themselves to address the state of the art and open problems in the scientific fields identified by the above-mentioned Projects [1, 2]. In carrying our this work, the WG1 strongly benefited from the participation of IDS Ingegneria dei Sistemi, one of the biggest GPR manufacturers, as well as from the contribution of external experts as David J. Daniels and Erica Utsi, sharing with the Action Members their wide experience on GPR technology and methodology (First General Meeting, July 2013). The synergy with WG2 and WG4 of the Action was useful for a deep understanding of the problems, merits and limits of available GPR equipment, as well as to discuss how to quantify the reliability of GPR results. An

  12. EMRE Is a Matrix Ca(2+) Sensor that Governs Gatekeeping of the Mitochondrial Ca(2+) Uniporter.

    PubMed

    Vais, Horia; Mallilankaraman, Karthik; Mak, Don-On Daniel; Hoff, Henry; Payne, Riley; Tanis, Jessica E; Foskett, J Kevin

    2016-01-26

    The mitochondrial uniporter (MCU) is an ion channel that mediates Ca(2+) uptake into the matrix to regulate metabolism, cell death, and cytoplasmic Ca(2+) signaling. Matrix Ca(2+) concentration is similar to that in cytoplasm, despite an enormous driving force for entry, but the mechanisms that prevent mitochondrial Ca(2+) overload are unclear. Here, we show that MCU channel activity is governed by matrix Ca(2+) concentration through EMRE. Deletion or charge neutralization of its matrix-localized acidic C terminus abolishes matrix Ca(2+) inhibition of MCU Ca(2+) currents, resulting in MCU channel activation, enhanced mitochondrial Ca(2+) uptake, and constitutively elevated matrix Ca(2+) concentration. EMRE-dependent regulation of MCU channel activity requires intermembrane space-localized MICU1, MICU2, and cytoplasmic Ca(2+). Thus, mitochondria are protected from Ca(2+) depletion and Ca(2+) overload by a unique molecular complex that involves Ca(2+) sensors on both sides of the inner mitochondrial membrane, coupled through EMRE.

  13. Ca isotopic fractionation patterns in forest ecosystems

    NASA Astrophysics Data System (ADS)

    Kurtz, A. C.; Takagi, K.

    2012-12-01

    Calcium stable isotope ratios are an emerging tracer of the biogeochemical cycle of Ca that are just beginning to see significant application to forest ecosystems. The primary source of isotopic fractionation in these systems is discrimination against light Ca during uptake by plant roots. Cycling of vegetation-fractionated Ca establishes isotopically distinct Ca pools within a forest ecosystem. In some systems, the shallow soil exchangeable Ca pool is isotopically heavy relative to Ca inputs. This has been explained by preferential removal of light Ca from the soil. In other systems, the soil exchange pool is isotopically light relative to inputs, which is explained by recycling of plant-fractionated light Ca back into soil. Thus vegetation uptake of light Ca has been called on to account for both isotopically heavy and light Ca in the shallow soil exchange pools. We interpret patterns in ecosystem δ44Ca with the aid of a simple box model of the forest Ca cycle. We suggest that the δ44Ca of exchangeable Ca in the shallow soil pool primarily reflects the relative magnitude of three key fluxes in a forest Ca cycle, 1) the flux of external Ca into the system via weathering or atmospheric deposition, 2) the uptake flux of Ca from soils into the vegetation pool, and 3) the return flux of Ca to shallow soils via remineralization of leaf litter. Two observations that emerge from our model may aid in the application of Ca isotopes to provide insight into the forest Ca cycle. First, regardless of the magnitude of both vegetation Ca uptake and isotopic fractionation, the δ44Ca of the soil exchange pool will equal the input δ44Ca unless the plant uptake and remineralization fluxes are out of balance. A second observation is that the degree to which the shallow soil exchange pool δ44Ca can differ from the input ratio is controlled by the relative rates of biological uptake and external Ca input. Significant differences between soil exchange and input δ44Ca are seen only

  14. Faculty: Thy Administrator's Keeper? Some Evidence

    ERIC Educational Resources Information Center

    Cunningham, Brendan M.

    2009-01-01

    Colleges and universities face a principal-agent problem. There are information asymmetries over the actions chosen by administrators. Because non-profit constraints limit the financial stake of trustees there may be insufficient monitoring of administrators and, consequentially, shirking. It is conceivable that faculty will serve as "delegated…

  15. Decoding Ca2+ signals in plants

    NASA Technical Reports Server (NTRS)

    Sathyanarayanan, P. V.; Poovaiah, B. W.

    2004-01-01

    Different input signals create their own characteristic Ca2+ fingerprints. These fingerprints are distinguished by frequency, amplitude, duration, and number of Ca2+ oscillations. Ca(2+)-binding proteins and protein kinases decode these complex Ca2+ fingerprints through conformational coupling and covalent modifications of proteins. This decoding of signals can lead to a physiological response with or without changes in gene expression. In plants, Ca(2+)-dependent protein kinases and Ca2+/calmodulin-dependent protein kinases are involved in decoding Ca2+ signals into phosphorylation signals. This review summarizes the elements of conformational coupling and molecular mechanisms of regulation of the two groups of protein kinases by Ca2+ and Ca2+/calmodulin in plants.

  16. Numerical modelling of GPR ground-matching enhancement by a chirped multilayer structure - output of cooperation within COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Baghdasaryan, Hovik V.; Knyazyan, Tamara M.; Hovhannisyan, Tamara. T.; Marciniak, Marian; Pajewski, Lara

    2016-04-01

    As is well know, Ground Penetrating Radar (GPR) is an electromagnetic technique for the detection and imaging of buried objects, with resolution ranging from centimeters to few meters [1, 2]. Though this technique is mature enough and different types of GPR devices are already in use, some problems are still waiting for their solution [3]. One of them is to achieve a better matching of transmitting GPR antenna to the ground, that will increase the signal penetration depth and the signal/noise ratio at the receiving end. In the current work, a full-wave electromagnetic modelling of the interaction of a plane wave with a chirped multilayered structure on the ground is performed, via numerical simulation. The method of single expression is used, which is a suitable technique for multi-boundary problems solution [4, 5]. The considered multilayer consists of two different dielectric slabs of low and high permittivity, where the highest value of permittivity doesn't exceed the permittivity of the ground. The losses in the ground are suitably taken into account. Two types of multilayers are analysed. Numerical results are obtained for the reflectance from the structure, as well as for the distributions of electric field components and power flow density in both the considered structures and the ground. The obtained results indicate that, for a better matching with the ground, the layer closer to the ground should be the high-permittivity one. Acknowledgement This work benefited from networking activities carried out within the EU funded COST Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar" (www.GPRadar.eu, www.cost.eu). Part of this work was developed during the Short-Term Scientific Mission COST-STSM-TU1208-25016, carried out by Prof. Baghdasaryan in the National Institute of Telecommunications in Warsaw, Poland. References [1] H. M. Jol. Ground Penetrating Radar: Theory and Applications. Elsevier, 2009. 509 pp. [2] R. Persico. Introduction to

  17. Turnip mosaic virus (TuMV) is able to use alleles of both eIF4E and eIF(iso)4E from multiple loci of the diploid Brassica rapa.

    PubMed

    Jenner, Carol E; Nellist, Charlotte F; Barker, Guy C; Walsh, John A

    2010-11-01

    Three copies of eIF4E and three copies of eIF(iso)4E have been identified and sequenced from a Turnip mosaic virus (TuMV)-susceptible, inbred, diploid Brassica rapa line, R-o-18. One of the copies of eIF4E lacked exons 2 and 3 and appeared to be a pseudogene. The two other copies of eIF4E and two of the three copies of eIF(iso)4E were isolated from a bacterial artificial chromosome library of R-o-18. Using an Arabidopsis line (Col-0::dSpm) with a transposon knock-out of the eIF(iso)4E gene which resulted in a change from complete susceptibility to complete resistance to TuMV, complementation experiments were carried out with the two versions of eIF4E and the two versions of eIF(iso)4E. When transformed into Col-0::dSpm, all four Brassica transgenes complemented the Arabidopsis eIF(iso)4E knock-out, conferring susceptibility to both mechanical and aphid challenge with TuMV. One of the copies of eIF4E did not appear to support viral replication as successfully as the other copy of eIF4E or the two copies of eIF(iso)4E. The results show that TuMV can use both eIF4E and eIF(iso)4E from B. rapa for replication and, for the first time, that a virus can use eIF4E and eIF(iso)4E from multiple loci of a single host plant.

  18. Turnip mosaic virus (TuMV) is able to use alleles of both eIF4E and eIF(iso)4E from multiple loci of the diploid Brassica rapa.

    PubMed

    Jenner, Carol E; Nellist, Charlotte F; Barker, Guy C; Walsh, John A

    2010-11-01

    Three copies of eIF4E and three copies of eIF(iso)4E have been identified and sequenced from a Turnip mosaic virus (TuMV)-susceptible, inbred, diploid Brassica rapa line, R-o-18. One of the copies of eIF4E lacked exons 2 and 3 and appeared to be a pseudogene. The two other copies of eIF4E and two of the three copies of eIF(iso)4E were isolated from a bacterial artificial chromosome library of R-o-18. Using an Arabidopsis line (Col-0::dSpm) with a transposon knock-out of the eIF(iso)4E gene which resulted in a change from complete susceptibility to complete resistance to TuMV, complementation experiments were carried out with the two versions of eIF4E and the two versions of eIF(iso)4E. When transformed into Col-0::dSpm, all four Brassica transgenes complemented the Arabidopsis eIF(iso)4E knock-out, conferring susceptibility to both mechanical and aphid challenge with TuMV. One of the copies of eIF4E did not appear to support viral replication as successfully as the other copy of eIF4E or the two copies of eIF(iso)4E. The results show that TuMV can use both eIF4E and eIF(iso)4E from B. rapa for replication and, for the first time, that a virus can use eIF4E and eIF(iso)4E from multiple loci of a single host plant. PMID:20672877

  19. CaFe interstellar clouds

    NASA Astrophysics Data System (ADS)

    Bondar, A.; Kozak, M.; Gnaciński, P.; Galazutdinov, G. A.; Beletsky, Y.; Krełowski, J.

    2007-07-01

    A new kind of interstellar cloud is proposed. These are rare (just a few examples among ~300 lines of sight) objects with the CaI 4227-Å, FeI 3720-Å and 3860-Å lines stronger than those of KI (near 7699 Å) and NaI (near 3302 Å). We propose the name `CaFe' for these clouds. Apparently they occupy different volumes from the well-known interstellar HI clouds where the KI and ultraviolet NaI lines are dominant features. In the CaFe clouds we have not found either detectable molecular features (CH, CN) or diffuse interstellar bands which, as commonly believed, are carried by some complex, organic molecules. We have found the CaFe clouds only along sightlines toward hot, luminous (and thus distant) objects with high rates of mass loss. In principle, the observed gas-phase interstellar abundances reflect the combined effects of the nucleosynthetic history of the material, the depletion of heavy elements into dust grains and the ionization state of these elements which may depend on irradiation by neighbouring stars. Based on data collected using the Maestro spectrograph at the Terskol 2-m telescope, Russia; and on data collected using the ESO Feros spectrograph; and on data obtained from the ESO Science Archive Facility acquired with the UVES spectrograph, Chile. E-mail: `arctur'@rambler.ru (AB); marizak@astri.uni.torun.pl (MK); pg@iftia.univ.gda.pl (PG); gala@boao.re.kr (GAG); ybialets@eso.org (YB); jacek@astri.uni.torun.pl (JK)

  20. Les aspects des frottis cervico-vaginaux chez les femmes vivants avec le VIH suivies à Thiès/Sénégal et association avec le degré d'immunodépression

    PubMed Central

    Bammo, Mariama; Dioussé, Pauline; Thiam, Marietou; Diop, Madoky Maguatte; Berthe, Adama; Faye, Flugence Abdou; Diallo, Thierno Abdoul Aziz; Sarr, Fatou Seck; Dione, Haby; Toure, Papa Souleymane; Diop, Bernard Marcel; Ka, Mamadou Mortalla

    2015-01-01

    De nombreuses études ont démontré que les femmes infectées par le VIH ont un risque accru de survenue de néoplasies cervicales intra épithéliales. L'association entre les deux affections étant bidirectionnelle, l'objectif était de décrire les anomalies cervicales chez les femmes séropositives au virus de l'immunodéficience humaine (VIH), de rechercher des facteurs associés et de proposer des recommandations en termes de suivi de ces femmes. Il s'agissait d'une étude transversale, multicentrique recensant l'ensemble des frottis cervico-vaginaux (FCV) et des colposcopies des patientes infectées par le VIH entre 2012 et 2014 dans les services de dermatologie de Thiès et de Mbour. Les données étaient recueillies et analysées par le logiciel EPI Info 2012 version 3.5.4. Les tests statistiques ont été effectués avec un seuil de significativité p <0,05. Etaient inclus 125 patientes. L’âge moyen était de 38,98 ± 10.2 ans [20-77]. Il n'y avait aucun signe d'appels dans 82.4%. Le FCV était normal dans 32.8%, inflammatoire dans 44.8%. Les anomalies cytologiques concernaient 22,4% dont, ASC-H (suspicion de lésions de haut grade: 2.4%), LSIL (lésions de bas grade: 8.8%), HSIL (lésions de haut grade: 4%). Leur majorité (60.7%) avaient un taux de CD4 < 500 et étaient au stade 3 de l'OMS dans 64.3%; la biopsie montrait une dysplasie sévère chez 37.5% des patientes ayant pu réaliser cet examen. Deux patientes ont bénéficié d'un traitement curatif notamment l'exérèse chirurgicale. La survenue de dysplasies cervicales même précoces semble être associée à un stade avancé de l'infection VIH. Un dépistage et un traitement précoces sont absolument nécessaires. PMID:26834915

  1. TU-A-12A-01: Consistency of Lung Expansion and Contraction During Respiration: Implications for Quantitative Imaging

    SciTech Connect

    Patton, T; Du, K; Bayouth, J; Christensen, G; Reinhardt, J

    2014-06-15

    lung tissue expansion. NIH Grant: R01 CA166703.

  2. Evolution of Seawater 44Ca/40Ca Through the Late Cretaceous and Cenozoic

    NASA Astrophysics Data System (ADS)

    Castillo, P. R.; Gopalan, K.; Norris, R. D.; MacIsaac, C.; Liu, X.; MacDougall, J. D.

    2009-12-01

    We analyzed the Ca concentrations and 44Ca/40Ca ratios of surface ocean planktonic (Morozovella, Acarinina, Dentoglobigerina) and benthic (Gavelinella) foraminifera of Late Cretaceous to Late Oligocene ages from DSDP and ODP sites in the Pacific, Atlantic and Indian oceans in order to fill a major gap in the Phanerozoic seawater 44Ca/40Ca curve (Farkass et al., Geochim. Cosmochim. Acta 71, 2007). Our new 44Ca/40Ca data indicate a general increase in foraminiferan-based seawater 44Ca/40Ca from ~-1.3 ‰ δ44Ca/40CaSW in Late Cretaceous to ~0.0 ‰ δ44Ca/40CaSW in Early Miocene (Heuser et al., Paleocean. 20, 2005; Sime et al., Geochim. Cosmochim. Acta 71, 2007). In detail, the 44Ca/40Ca ratio stepped abruptly from ~-1.3 ‰ δ44Ca/40CaSW to a slightly higher value of ~-1.1 ‰ δ44Ca/40CaSW across the Cretaceous-Tertiary (K/T) boundary. A slight positive excursion of ~0.2 ‰ above the background value occurred after the Paleocene Thermal Maximum (55 Ma) but otherwise, the Paleocene to Middle Eocene ratio is relatively stable at ~-1.0 ‰ δ44Ca/40CaSW. The most prominent increase in foraminiferan-based seawater 44Ca/40Ca occurred from Late Eocene to Late Oligocene, roughly coincident with the initial phase of the rapid and steady rise of marine carbonate 87Sr/86Sr ratio in the Tertiary (e.g., DePaolo and Ingram, Science 227, 1985).

  3. Fine tuning of cytosolic Ca 2+ oscillations

    PubMed Central

    Dupont, Geneviève; Combettes, Laurent

    2016-01-01

    Ca 2+ oscillations, a widespread mode of cell signaling, were reported in non-excitable cells for the first time more than 25 years ago. Their fundamental mechanism, based on the periodic Ca 2+ exchange between the endoplasmic reticulum and the cytoplasm, has been well characterized. However, how the kinetics of cytosolic Ca 2+ changes are related to the extent of a physiological response remains poorly understood. Here, we review data suggesting that the downstream targets of Ca 2+ are controlled not only by the frequency of Ca 2+ oscillations but also by the detailed characteristics of the oscillations, such as their duration, shape, or baseline level. Involvement of non-endoplasmic reticulum Ca 2+ stores, mainly mitochondria and the extracellular medium, participates in this fine tuning of Ca 2+ oscillations. The main characteristics of the Ca 2+ exchange fluxes with these compartments are also reviewed.

  4. [Effect of polycarbophil Ca on IBS].

    PubMed

    Mine, Tetsuya

    2006-08-01

    In this chapter, I mentioned the effect of polycarbophil Ca on IBS. IBS is classified into 3 types; diarrhea type, constipation type and combined type. Polycarbophil Ca is effective for all types of IBS.

  5. Fine tuning of cytosolic Ca (2+) oscillations.

    PubMed

    Dupont, Geneviève; Combettes, Laurent

    2016-01-01

    Ca (2+) oscillations, a widespread mode of cell signaling, were reported in non-excitable cells for the first time more than 25 years ago. Their fundamental mechanism, based on the periodic Ca (2+) exchange between the endoplasmic reticulum and the cytoplasm, has been well characterized. However, how the kinetics of cytosolic Ca (2+) changes are related to the extent of a physiological response remains poorly understood. Here, we review data suggesting that the downstream targets of Ca (2+) are controlled not only by the frequency of Ca (2+) oscillations but also by the detailed characteristics of the oscillations, such as their duration, shape, or baseline level. Involvement of non-endoplasmic reticulum Ca (2+) stores, mainly mitochondria and the extracellular medium, participates in this fine tuning of Ca (2+) oscillations. The main characteristics of the Ca (2+) exchange fluxes with these compartments are also reviewed. PMID:27630768

  6. Fine tuning of cytosolic Ca 2+ oscillations

    PubMed Central

    Dupont, Geneviève; Combettes, Laurent

    2016-01-01

    Ca 2+ oscillations, a widespread mode of cell signaling, were reported in non-excitable cells for the first time more than 25 years ago. Their fundamental mechanism, based on the periodic Ca 2+ exchange between the endoplasmic reticulum and the cytoplasm, has been well characterized. However, how the kinetics of cytosolic Ca 2+ changes are related to the extent of a physiological response remains poorly understood. Here, we review data suggesting that the downstream targets of Ca 2+ are controlled not only by the frequency of Ca 2+ oscillations but also by the detailed characteristics of the oscillations, such as their duration, shape, or baseline level. Involvement of non-endoplasmic reticulum Ca 2+ stores, mainly mitochondria and the extracellular medium, participates in this fine tuning of Ca 2+ oscillations. The main characteristics of the Ca 2+ exchange fluxes with these compartments are also reviewed. PMID:27630768

  7. Tu Amigo Pepe: Evaluation of a Multi-media Marketing Campaign that Targets Young Latino Immigrant MSM with HIV Testing Messages.

    PubMed

    Solorio, Rosa; Norton-Shelpuk, Pamela; Forehand, Mark; Montaño, Daniel; Stern, Joshua; Aguirre, Joel; Martinez, Marcos

    2016-09-01

    Latino immigrant men who have sex with men (MSM) are at risk for HIV and delayed diagnosis in the United States. This paper describes the evaluation of a pilot of the Tu Amigo Pepe, a multimedia HIV testing campaign aimed at Latino MSM in Seattle, WA particularly targeting immigrants who may not identify as gay, ages 18-30 years old. The 16-week campaign included Spanish-language radio public service announcements (PSAs), a Web site, social media outreach, a reminder system using mobile technology, print materials and a toll-free hotline. In developing the PSAs, the Integrated Behavioral Model was used as a framework to reframe negative attitudes, beliefs and norms towards HIV testing with positive ones as well as to promote self-efficacy towards HIV testing. The campaign had a significant and immediate impact on attitudes, beliefs, norms and self-efficacy towards HIV testing as well as on actual behavior, with HIV testing rates increasing over time. PMID:26850101

  8. Tu Amigo Pepe: Evaluation of a Multi-media Marketing Campaign that Targets Young Latino Immigrant MSM with HIV Testing Messages.

    PubMed

    Solorio, Rosa; Norton-Shelpuk, Pamela; Forehand, Mark; Montaño, Daniel; Stern, Joshua; Aguirre, Joel; Martinez, Marcos

    2016-09-01

    Latino immigrant men who have sex with men (MSM) are at risk for HIV and delayed diagnosis in the United States. This paper describes the evaluation of a pilot of the Tu Amigo Pepe, a multimedia HIV testing campaign aimed at Latino MSM in Seattle, WA particularly targeting immigrants who may not identify as gay, ages 18-30 years old. The 16-week campaign included Spanish-language radio public service announcements (PSAs), a Web site, social media outreach, a reminder system using mobile technology, print materials and a toll-free hotline. In developing the PSAs, the Integrated Behavioral Model was used as a framework to reframe negative attitudes, beliefs and norms towards HIV testing with positive ones as well as to promote self-efficacy towards HIV testing. The campaign had a significant and immediate impact on attitudes, beliefs, norms and self-efficacy towards HIV testing as well as on actual behavior, with HIV testing rates increasing over time.

  9. Incidence and Remission of Parasomnias among Adolescent Children in the Tucson Children’s Assessment of Sleep Apnea (TuCASA) Study

    PubMed Central

    Furet, Oscar; Goodwin, James L.; Quan, Stuart F.

    2011-01-01

    Background Longitudinal assessments of parasomnias in the adolescent population are scarce. This analysis aims to identify the incidence and remission of parasomnias in the adolescent age group. Methods The TuCASA study is a prospective cohort study that initially enrolled children between the ages of 6 and 11 years (Time 1) and subsequently restudied them approximately 5 years later (Time 2). At both time points parents were asked to complete a comprehensive sleep habits questionnaire designed to assess the severity of sleep-related symptoms that included questions about enuresis (EN), sleep terrors (TR), sleep walking (SW) and sleep talking (ST). Results There were 350 children participating at Time 1 who were studied as adolescents at time 2. The mean interval between measurements was (4.6 years). The incidence of EN, TR, ST, and SW in these 10–18 year old children was 0.3%, 0.6%, 6.0% and 1.1% respectively. Remission rates were 70.8%, 100%, 64.8% and 50.0% respectively. Conclusions The incidence rates of EN, TR, and SW were relatively low moving from childhood to adolescence while remission rates were high across all parasomnias. PMID:21847446

  10. Effectiveness: N(sub TU) relationships for the design and performance evaluation of additional shell-and-tube heat exchanger geometries

    NASA Astrophysics Data System (ADS)

    1988-11-01

    This Data Item 88021, an addition to the Sub-series on Heat Transfer, complements ESDU 86018 by extending the range of configurations covered there and in particular considering the effect of using small numbers of baffles for E- and J-shells and the use of J-shells in series. It also explores the limitations of the assumptions associated with the effectiveness - N(sub TU) method and shows where those assumptions break down. The curves presented for each exchanger geometry show the locus of designs for which a temperature cross may occur and the locus of 95 percent heat transfer effectiveness which indicates the region of uneconomic design. The method assumes a linear temperature/enthalpy relationship (constant specific heat capacity) for both streams. It applies to boiling or condensing flow of a single component with no temperature change, or boiling and condensing flow of a mixture that is always two-phase. It excludes conditions in which transition from single- to two-phase flow occurs. However, by use of average property values, it is possible to extend the method to apply to cases where there is some variation of physical and thermodynamic properties with temperature.

  11. Modeling of Turbulent Boundary Layer Surface Pressure Fluctuation Auto and Cross Spectra - Verification and Adjustments Based on TU-144LL Data

    NASA Technical Reports Server (NTRS)

    Rackl, Robert; Weston, Adam

    2005-01-01

    The literature on turbulent boundary layer pressure fluctuations provides several empirical models which were compared to the measured TU-144 data. The Efimtsov model showed the best agreement. Adjustments were made to improve its agreement further, consisting of the addition of a broad band peak in the mid frequencies, and a minor modification to the high frequency rolloff. The adjusted Efimtsov predicted and measured results are compared for both subsonic and supersonic flight conditions. Measurements in the forward and middle portions of the fuselage have better agreement with the model than those from the aft portion. For High Speed Civil Transport supersonic cruise, interior levels predicted by use of this model are expected to increase by 1-3 dB due to the adjustments to the Efimtsov model. The space-time cross-correlations and cross-spectra of the fluctuating surface pressure were also investigated. This analysis is an important ingredient in structural acoustic models of aircraft interior noise. Once again the measured data were compared to the predicted levels from the Efimtsov model.

  12. Determinants in CaV1 Channels That Regulate the Ca2+ Sensitivity of Bound Calmodulin*

    PubMed Central

    Halling, D. Brent; Georgiou, Dimitra K.; Black, D. J.; Yang, Guojun; Fallon, Jennifer L.; Quiocho, Florante A.; Pedersen, Steen E.; Hamilton, Susan L.

    2009-01-01

    Calmodulin binds to IQ motifs in the α1 subunit of CaV1.1 and CaV1.2, but the affinities of calmodulin for the motif and for Ca2+ are higher when bound to CaV1.2 IQ. The CaV1.1 IQ and CaV1.2 IQ sequences differ by four amino acids. We determined the structure of calmodulin bound to CaV1.1 IQ and compared it with that of calmodulin bound to CaV1.2 IQ. Four methionines in Ca2+-calmodulin form a hydrophobic binding pocket for the peptide, but only one of the four nonconserved amino acids (His-1532 of CaV1.1 and Tyr-1675 of CaV1.2) contacts this calmodulin pocket. However, Tyr-1675 in CaV1.2 contributes only modestly to the higher affinity of this peptide for calmodulin; the other three amino acids in CaV1.2 contribute significantly to the difference in the Ca2+ affinity of the bound calmodulin despite having no direct contact with calmodulin. Those residues appear to allow an interaction with calmodulin with one lobe Ca2+-bound and one lobe Ca2+-free. Our data also provide evidence for lobe-lobe interactions in calmodulin bound to CaV1.2. PMID:19473981

  13. Autonomous CaMKII requires further stimulation by Ca2+/calmodulin for enhancing synaptic strength

    PubMed Central

    Barcomb, Kelsey; Buard, Isabelle; Coultrap, Steven J.; Kulbe, Jacqueline R.; O'Leary, Heather; Benke, Timothy A.; Bayer, K. Ulrich

    2014-01-01

    A hallmark feature of Ca2+/calmodulin (CaM)-dependent protein kinase II (CaMKII) is generation of autonomous (Ca2+-independent) activity by T286 autophosphorylation. Biochemical studies have shown that “autonomous” CaMKII is ∼5-fold further stimulated by Ca2+/CaM, but demonstration of a physiological function for such regulation within cells has remained elusive. In this study, CaMKII-induced enhancement of synaptic strength in rat hippocampal neurons required both autonomous activity and further stimulation. Synaptic strength was decreased by CaMKIIα knockdown and rescued by reexpression, but not by mutants impaired for autonomy (T286A) or binding to NMDA-type glutamate receptor subunit 2B (GluN2B; formerly NR2B; I205K). Full rescue was seen with constitutively autonomous mutants (T286D), but only if they could be further stimulated (additional T305/306A mutation), and not with two other mutations that additionally impair Ca2+/CaM binding. Compared to rescue with wild-type CaMKII, the CaM-binding-impaired mutants even had reduced synaptic strength. One of these mutants (T305/306D) mimicked an inhibitory autophosphorylation of CaMKII, whereas the other one (Δstim) abolished CaM binding without introducing charged residues. Inhibitory T305/306 autophosphorylation also reduced GluN2B binding, but this effect was independent of reduced Ca2+/CaM binding and was not mimicked by T305/306D mutation. Thus, even autonomous CaMKII activity must be further stimulated by Ca2+/CaM for enhancement of synaptic strength.—Barcomb, K., Buard, I., Coultrap, S. J., Kulbe, J. R., O'Leary, H., Benke, T. A., Bayer, K. U. Autonomous CaMKII requires further stimulation by Ca2+/calmodulin for enhancing synaptic strength. PMID:24843070

  14. TU-A-9A-07: X-Ray Acoustic Computed Tomography (XACT): 100% Sensitivity to X-Ray Absorption

    SciTech Connect

    Xiang, L; Ahmad, M; Nikoozadeh, A; Pratx, G; Khuri-Yakub, B; Xing, L

    2014-06-15

    gratefully acknowledge the Department of Defense Prostate Cancer Research Programs W81XWH-13-1-0481 (LX), the National Institutes of Health 1R01 CA133474 and 1R21 A153587, and SRFDP (20124407120012) for funding.

  15. TU-C-BRE-02: A Novel, Highly Efficient and Automated Quality Assurance Tool for Modern Linear Accelerators

    SciTech Connect

    Goddu, S; Sun, B; Yaddanapudi, S; Kamal, G; Mutic, S; Baltes, C; Rose, S; Stinson, K

    2014-06-15

    for QA of TrueBeam systems and its automation makes it highly efficient for testing both geometric and dosimetric aspects of the machine. This is very important for hypo-fractionated SBRT treatments. Received support from Varian Medical Systems, Palo Alto, CA 94304-1038.

  16. Controls on Sr/Ca in Scleractinian Corals: The Effects of Ca-ATPase and Ca channels on Skeletal Chemistry

    NASA Astrophysics Data System (ADS)

    Allison, N.; Cohen, I.; Finch, A. A.; Erez, J.

    2010-12-01

    Coral skeletal Sr/Ca is a commonly used palaeothermometer and has been used to estimate past sea surface temperatures. However the processes controlling Sr incorporation in coral aragonite are poorly understood. The Sr/Ca chemistry of the massive Porites spp. corals typically used for palaeoenvironmental reconstruction is dominated by short-term (weekly-monthly) oscillations of ~10% which do not reflect seawater temperature. This heterogeneity may reflect variations in the composition of the fluid used for calcification. Coral skeletons precipitate from an extracellular calcifying fluid enclosed in a semi-isolated space between the skeleton and the calicoblastic epithelium (the tissue layer at the base of the coral organism). Seawater diffuses directly to the calcification site and the calcification fluid has a composition derived from that of seawater but modified by other transport processes. In zooxanthellate corals, Ca2+ is transported transcellularly to the calcification site by both calcium channels and by the carrier protein Ca-ATPase. Sr2+ has a similar ionic radius to Ca2+, but it is not clear if Sr2+ can substitute for Ca2+ in these transport mechanisms. Variations in the relative contributions of each of the transport mechanisms to the calcification fluid and the efficiencies with which each process transports Sr2+ and Ca2+ could explain the Sr/Ca heterogeneity observed in coral skeletons. To test the impact of transcellular Ca transport processes on skeletal Sr/Ca and Mg/Ca we cultured Pocillopora damicornis corals in the presence of inhibitors of Ca-ATPase (ruthenium red) and Ca channels (verapamil). The photosynthesis, respiration and calcification rates of the colonies were monitored throughout the experiment. The skeleton subsequently deposited was identified (by 42Ca spike) and analysed by secondary ion mass spectrometry. The Sr/Ca and Mg/Ca of the aragonite deposited in the presence of either of the inhibitors was not significantly different from

  17. Mission CaMKIIγ: shuttle calmodulin from membrane to nucleus.

    PubMed

    Malik, Zulfiqar A; Stein, Ivar S; Navedo, Manuel F; Hell, Johannes W

    2014-10-01

    Neuronal plasticity depends on plasma membrane Ca(2+) influx, resulting in activity-dependent gene transcription. Calmodulin (CaM) activated by Ca(2+) initiates the nuclear events, but how CaM makes its way to the nucleus has remained elusive. Ma et al. now show that CaMKIIγ transports CaM from cell surface Ca(2+) channels to the nucleus.

  18. Ca2+ Cycling in Heart Failure

    PubMed Central

    Luo, Min; Anderson, Mark E.

    2013-01-01

    Ca2+ plays a crucial role in connecting membrane excitability with contraction in myocardium. The hallmark features of heart failure are mechanical dysfunction and arrhythmias; defective intracellular Ca2+ homeostasis is a central cause of contractile dysfunction and arrhythmias in failing myocardium. Defective Ca2+ homeostasis in heart failure can result from pathological alteration in the expression and activity of an increasingly understood collection of Ca2+ homeostatic binding proteins, ion channels and enzymes. This review focuses on the molecular mechanisms of defective Ca2+ cycling in heart failure and consider how fundamental understanding of these pathways may translate into novel and innovative therapies. PMID:23989713

  19. Distinct Roles for Dorsal CA3 and CA1 in Memory for Sequential Nonspatial Events

    ERIC Educational Resources Information Center

    Farovik, Anja; Dupont, Laura M.; Eichenbaum, Howard

    2010-01-01

    Previous studies have suggested that dorsal hippocampal areas CA3 and CA1 are both involved in representing sequences of events that compose unique episodes. However, it is uncertain whether the contribution of CA3 is restricted to spatial information, and it is unclear whether CA1 encodes order per se or contributes by an active maintenance of…

  20. Expression and Localization of CaBP Ca2+ Binding Proteins in the Mouse Cochlea.

    PubMed

    Yang, Tian; Scholl, Elizabeth S; Pan, Ning; Fritzsch, Bernd; Haeseleer, Françoise; Lee, Amy

    2016-01-01

    CaBPs are a family of EF-hand Ca2+ binding proteins that are structurally similar to calmodulin. CaBPs can interact with, and yet differentially modulate, effectors that are regulated by calmodulin, such as Cav1 voltage-gated Ca2+ channels. Immunolabeling studies suggest that multiple CaBP family members (CaBP1, 2, 4, and 5) are expressed in the cochlea. To gain insights into the respective auditory functions of these CaBPs, we characterized the expression and cellular localization of CaBPs in the mouse cochlea. By quantitative reverse transcription PCR, we show that CaBP1 and CaBP2 are the major CaBPs expressed in mouse cochlea both before and after hearing onset. Of the three alternatively spliced variants of CaBP1 (caldendrin, CaBP1-L, and CaBP1-S) and CaBP2 (CaBP2-alt, CaBP2-L, CaBP2-S), caldendrin and CaBP2-alt are the most abundant. By in situ hybridization, probes recognizing caldendrin strongly label the spiral ganglion, while probes designed to recognize all three isoforms of CaBP1 weakly label both the inner and outer hair cells as well as the spiral ganglion. Within the spiral ganglion, caldendrin/CaBP1 labeling is associated with cells resembling satellite glial cells. CaBP2-alt is strongly expressed in inner hair cells both before and after hearing onset. Probes designed to recognize all three variants of CaBP2 strongly label inner hair cells before hearing onset and outer hair cells after the onset of hearing. Thus, CaBP1 and CaBP2 may have overlapping roles in regulating Ca2+ signaling in the hair cells, and CaBP1 may have an additional function in the spiral ganglion. Our findings provide a framework for understanding the role of CaBP family members in the auditory periphery. PMID:26809054

  1. Non–Ca2+-conducting Ca2+ channels in fish skeletal muscle excitation-contraction coupling

    PubMed Central

    Schredelseker, Johann; Shrivastav, Manisha; Dayal, Anamika; Grabner, Manfred

    2010-01-01

    During skeletal muscle excitation-contraction (EC) coupling, membrane depolarizations activate the sarcolemmal voltage-gated L-type Ca2+ channel (CaV1.1). CaV1.1 in turn triggers opening of the sarcoplasmic Ca2+ release channel (RyR1) via interchannel protein–protein interaction to release Ca2+ for myofibril contraction. Simultaneously to this EC coupling process, a small and slowly activating Ca2+ inward current through CaV1.1 is found in mammalian skeletal myotubes. The role of this Ca2+ influx, which is not immediately required for EC coupling, is still enigmatic. Interestingly, whole-cell patch clamp experiments on freshly dissociated skeletal muscle myotubes from zebrafish larvae revealed the lack of such Ca2+ currents. We identified two distinct isoforms of the pore-forming CaV1.1α1S subunit in zebrafish that are differentially expressed in superficial slow and deep fast musculature. Both do not conduct Ca2+ but merely act as voltage sensors to trigger opening of two likewise tissue-specific isoforms of RyR1. We further show that non-Ca2+ conductivity of both CaV1.1α1S isoforms is a common trait of all higher teleosts. This non-Ca2+ conductivity of CaV1.1 positions teleosts at the most-derived position of an evolutionary trajectory. Though EC coupling in early chordate muscles is activated by the influx of extracellular Ca2+, it evolved toward CaV1.1-RyR1 protein–protein interaction with a relatively small and slow influx of external Ca2+ in tetrapods. Finally, the CaV1.1 Ca2+ influx was completely eliminated in higher teleost fishes. PMID:20212109

  2. Adenosine stimulates Ca2+ fluxes and increases cytosolic free Ca2+ in cultured rat mesangial cells.

    PubMed Central

    Olivera, A; López-Rivas, A; López-Novoa, J M

    1992-01-01

    Adenosine has been associated with cellular Ca2+ metabolism in some cell types. Since adenosine is able to contract glomerular mesangial cells in culture, and since Ca2+ is the main messenger mediating contractile responses, we studied the effect of adenosine on 45Ca2+ movements into and out of mesangial cells and on the cytosolic free Ca2+ concentration ([Ca2+]i). Adenosine at 0.1 mM increased 45Ca2+ uptake (basal, 9993 +/- 216; + adenosine, 14823 +/- 410 d.p.m./mg; P less than 0.01) through verapamil-sensitive Ca2+ channels. These channels seem to be of the A1-adenosine receptor subtype. Adenosine also stimulated 45Ca2+ efflux from 45Ca(2+)-loaded mesangial cells. This effect was accompanied by a net depletion of intracellular 45Ca2+ content under isotopic equilibrium conditions (basal, 24213 +/- 978; + adenosine, 18622 +/- 885 d.p.m./mg; P less than 0.05). The increase in 45Ca2+ efflux was inhibited by a Ca(2+)-free medium or in the presence of 10 microM-verapamil. However, the intracellular Ca(2+)-release blocker TMB-8 (10 microM) only partially inhibited the adenosine-stimulated 45Ca2+ efflux. In addition, adenosine induced an elevation in [Ca2+]i in mesangial cells with an initial transient peak within 15 s (basal, 113 +/- 7; adenosine, 345 +/- 46 nM), and a secondary increase which was slower (3-4 min) and of lower magnitude than the initial peak (250 +/- 21 nM). In summary, adenosine elevates [Ca2+]i and stimulates both Ca2+ uptake from the extracellular pool and Ca2+ efflux from intracellular pools in mesangial cells. The Ca2+ release from internal stores is produced by a combination of a TMB-8-inhibitable and a non-TMB-8-inhibitable mechanism, and seems to be dependent on Ca2+ influx. PMID:1554371

  3. GPR surveying of transport infrastructures and buildings; underground utility and void sensing - ongoing activities in Working Group 2 of COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Pajewski, Lara; Plati, Christina; Derobert, Xavier

    2015-04-01

    This work aims at presenting the ongoing research activities carried out in Working Group 2 'GPR surveying of pavements, bridges, tunnels and buildings; underground utility and void sensing' of the COST (European COoperation in Science and Technology) Action TU1208 'Civil Engineering Applications of Ground Penetrating Radar' (www.GPRadar.eu). The principal goal of the COST Action TU1208 is to exchange and increase scientific-technical knowledge and experience of Ground Penetrating Radar (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. Four Working Groups (WGs) carry out the research activities. WG1 focuses on the development of innovative GPR equipment dedicated for civil engineering applications. WG2 deals with the development of guidelines and protocols for the surveying, through the use of a GPR system, of transport infrastructure and buildings, as well as for the sensing of utilities and voids. WG3 deals with the development of electromagnetic forward and inverse scattering methods, for the characterization of GPR scenarios, as well as with data- processing algorithms for the elaboration of the data collected during GPR surveys. WG4 is concerned with the use of GPR in fields different from the civil engineering, as well as with the integration of GPR with other non-destructive testing techniques. Each WG includes several Projects. WG2 includes five Projects. Project 2.1 focuses on outlining 'Innovative inspection procedures for effective GPR surveying of critical transport infrastructures (pavements, bridges and tunnels).' Project 2.2 is concerned with the development of 'Innovative inspection procedures for effective GPR surveying of buildings.' Project 2.3 deals with identifying 'Innovative inspection procedures for effective GPR sensing and mapping of underground utilities and voids, with a focus to urban

  4. Comparative study of 64Cu/NOTA-[D-Tyr6,βAla11,Thi13,Nle14]BBN(6-14) monomer and dimers for prostate cancer PET imaging

    PubMed Central

    2012-01-01

    Background Gastrin-releasing peptide receptors [GRPR] are highly over-expressed in multiple cancers and have been studied as a diagnostic target. Multimeric gastrin-releasing peptides are expected to have enhanced tumor uptake and affinity for GRPR. In this study, a 64Cu-labeled 1,4,7-triazacyclononane-1,4,7-triacetic acid [NOTA]-monomer and two NOTA-dimers of [D-Tyr6,βAla11, Thi13, Nle14]bombesin(6-14) ] [BBN(6-14)] were compared. Methods Monomeric and dimeric peptides were synthesized on solid phase support and radiolabeled with 64Cu. NOTA-dimer 1 consists of asymmetrically linked BBN(6-14), while NOTA-dimer 2 has similar spacer between the two BBN(6-14) ligands and the chelator. In vitro GRPR-binding affinities were determined with competitive binding assays on PC3 human prostate cancer cells. In vivo stability and biodistribution of radiolabeled compounds were assessed in Balb/c mice. Cellular uptake and efflux were measured with radiolabeled NOTA-monomer and NOTA-dimer 2 on PC3 cells for up to 4 h. In vivo biodistribution kinetics were measured in PC3 tumor-bearing Balb/c nude mice by μ-positron emission tomography [μPET] imaging and confirmed by dissection and counting. Results NOTA-monomer, NOTA-dimers 1 and 2 were prepared with purity of 99%. The inhibition constants of the three BBN peptides were comparable and in the low nanomolar range. All 64Cu-labeled peptides were stable up to 24 h in mouse plasma and 1 h in vivo. 64Cu/NOTA-dimer 2 featuring a longer spacer between the two BBN(6-14) ligands is a more potent GRPR-targeting probe than 64Cu/NOTA-dimer 1. PC3 tumor uptake profiles are slightly different for 64Cu/NOTA-monomer and 64Cu/NOTA-dimer 2; the monomeric BBN-peptide tracer exhibited higher tumor uptake during the first 0.5 h and a fast renal clearance resulting in higher tumor-to-muscle ratio when compared to 64Cu/NOTA-dimer 2. The latter exhibited higher tumor-to-blood ratio and was retained longer at the tumor site when compared to 64Cu

  5. Ca isotope cycling in a forested ecosystem

    NASA Astrophysics Data System (ADS)

    Holmden, Chris; Bélanger, Nicolas

    2010-02-01

    Reports of large Ca isotope fractionations between trees and soils prompted this study of a Boreal forest ecosystem near La Ronge, Saskatchewan, to improve understanding of this phenomenon. The results on five tree species (black spruce, trembling aspen, white spruce, jack pine, balsam poplar) confirm that nutrient Ca uptake by plants favors the light isotopes, thus driving residual Ca in plant available soil pools towards enrichment in the heavy isotopes. Substantial within-tree fraction occurs in tissues formed along the transpiration stream, with low δ 44Ca values in fine roots (2 mm), intermediate values in stemwood, and high values in foliage. Separation factors between different plant tissues are similar between species, but the initial fractionation step in the tips of the fine roots is species specific, and/or sensitive to the local soil environment. Soil water δ 44Ca values appear to increase with depth to at least 35 cm below the top of the forest floor, which is close to the deepest level of fine roots. The heavy plant fractionated signature of Ca in the finely rooted upper soils filters downward where it is retained on ion exchange sites, leached into groundwater, and discharged into surface waters. The relationship between Ca uptake by tree fine roots and the pattern of δ 44Ca enrichment with soil depth was modeled for two Ca pools: the forest floor (litter) and the underlying (upper B) mineral soil. Six study plots were investigated along two hillside toposequences trending upwards from a first order stream. We used allometric equations describing the Ca distribution in boreal tree species to calculate weighted average δ 44Ca values for the stands in each plot and estimate Ca uptake rates. The δ 44Ca value of precipitation was measured, and soil weathering signatures deduced, by acid leaching of lower B mineral soils. Steady state equations were used to derive a set of model Ca fluxes and fractionation factors for each plot. The model reproduces

  6. Structure of the E. coli ribosome-EF-Tu complex at <3 Å resolution by Cs-corrected cryo-EM.

    PubMed

    Fischer, Niels; Neumann, Piotr; Konevega, Andrey L; Bock, Lars V; Ficner, Ralf; Rodnina, Marina V; Stark, Holger

    2015-04-23

    Single particle electron cryomicroscopy (cryo-EM) has recently made significant progress in high-resolution structure determination of macromolecular complexes due to improvements in electron microscopic instrumentation and computational image analysis. However, cryo-EM structures can be highly non-uniform in local resolution and all structures available to date have been limited to resolutions above 3 Å. Here we present the cryo-EM structure of the 70S ribosome from Escherichia coli in complex with elongation factor Tu, aminoacyl-tRNA and the antibiotic kirromycin at 2.65-2.9 Å resolution using spherical aberration (Cs)-corrected cryo-EM. Overall, the cryo-EM reconstruction at 2.9 Å resolution is comparable to the best-resolved X-ray structure of the E. coli 70S ribosome (2.8 Å), but provides more detailed information (2.65 Å) at the functionally important ribosomal core. The cryo-EM map elucidates for the first time the structure of all 35 rRNA modifications in the bacterial ribosome, explaining their roles in fine-tuning ribosome structure and function and modulating the action of antibiotics. We also obtained atomic models for flexible parts of the ribosome such as ribosomal proteins L9 and L31. The refined cryo-EM-based model presents the currently most complete high-resolution structure of the E. coli ribosome, which demonstrates the power of cryo-EM in structure determination of large and dynamic macromolecular complexes.

  7. Drugs preventing Na+ and Ca2+ overload.

    PubMed

    Ravens, U; Himmel, H M

    1999-03-01

    Cardiac intracellular Na+and Ca2+homeostasis is regulated by the concerted action of ion channels, pumps and exchangers. The Na+, K+-ATPase produces the electrochemical concentration gradient for Na+, which is the driving force for Ca2+removal from the cytosol via the Na+/Ca2+exchange. Reduction of this gradient by increased intracellular Na+concentration leads to cellular Ca2+overload resulting in arrhythmias and contractile dysfunction. Na+and Ca2+overload-associated arrhythmias can be produced experimentally by inhibition of Na+efflux (digitalis-induced intoxication) and by abnormal Na+influx via modulated Na+channels (veratridine, DPI 201-106; hypoxia) or via the Na+, H+exchanger. Theoretically, blockers of Na+and Ca2+channels, inhibitors of abnormal oscillatory release of Ca2+from internal stores or modulators of the Na+, Ca2+and Na+, H+exchanger activities could protect against cellular Na+and Ca2+overload. Three exemplary drugs that prevent Na+and Ca2+overload, i.e. the benzothiazolamine R56865, the methylenephenoxydioxy-derivative CP-060S, and the benzoyl-guanidine Hoe 642, a Na+, H+exchange blocker, are briefly reviewed with respect to their efficacy on digitalis-, veratridine- and ischaemia/reperfusion-induced arrhythmias. PMID:10094840

  8. Neuronal Ca2+ disregulation in diabetes mellitus.

    PubMed

    Biessels, Geert Jan; ter Laak, Mariël P; Hamers, Frank P T; Gispen, Willem Hendrik

    2002-07-01

    The Ca(2+) hypothesis of brain ageing and dementia may account for part of the available data on the pathogenesis of dementia and certain neurodegenerative disorders. The hypothesis proposes that disturbances in the homeostasis of neuronal cytosolic free Ca(2+) are part of a final common pathway, ultimately leading to neuronal dysfunction and cell death. The hypothesis also proposes that a small change in cytosolic free Ca(2+) sustained over a long period of time will result in similar damage as a large change over a short period. Diabetes mellitus is associated with neurological complications in the peripheral and central nervous system, as reflected in peripheral neuropathy, modest cognitive impairments and an increased risk of dementia. In animal models of diabetes, learning impairments are associated with alterations in Ca(2+) -dependent forms of hippocampal synaptic plasticity. Disturbances in the homeostasis of cytosolic free Ca(2+) may present a final common pathway in the multifactorial pathogenesis of neurological complications of diabetes, which involves vascular changes, oxidative stress, and non-enzymatic protein glycation. In line with the Ca(2+) hypothesis of neurodegenerative disorders, a prolonged, small increase in basal cytosolic Ca(2+) levels indeed exists in sensory neurones of diabetic animals. In addition, Ca(2+) dynamics are affected. Ca(2+) channel blockers, such as nimodipine, have been shown to improve experimental peripheral neuropathy, through a vascular mechanism, possibly in combination with direct neuronal effects. Preliminary studies indicate that nimodipine may also improve Ca(2+)-dependent forms of synaptic plasticity in the hippocampus of diabetic rats.

  9. The influence of Ca²⁺ buffers on free [Ca²⁺] fluctuations and the effective volume of Ca²⁺ microdomains.

    PubMed

    Weinberg, Seth H; Smith, Gregory D

    2014-06-17

    Intracellular calcium (Ca(2+)) plays a significant role in many cell signaling pathways, some of which are localized to spatially restricted microdomains. Ca(2+) binding proteins (Ca(2+) buffers) play an important role in regulating Ca(2+) concentration ([Ca(2+)]). Buffers typically slow [Ca(2+)] temporal dynamics and increase the effective volume of Ca(2+) domains. Because fluctuations in [Ca(2+)] decrease in proportion to the square-root of a domain's physical volume, one might conjecture that buffers decrease [Ca(2+)] fluctuations and, consequently, mitigate the significance of small domain volume concerning Ca(2+) signaling. We test this hypothesis through mathematical and computational analysis of idealized buffer-containing domains and their stochastic dynamics during free Ca(2+) influx with passive exchange of both Ca(2+) and buffer with bulk concentrations. We derive Langevin equations for the fluctuating dynamics of Ca(2+) and buffer and use these stochastic differential equations to determine the magnitude of [Ca(2+)] fluctuations for different buffer parameters (e.g., dissociation constant and concentration). In marked contrast to expectations based on a naive application of the principle of effective volume as employed in deterministic models of Ca(2+) signaling, we find that mobile and rapid buffers typically increase the magnitude of domain [Ca(2+)] fluctuations during periods of Ca(2+) influx, whereas stationary (immobile) Ca(2+) buffers do not. Also contrary to expectations, we find that in the absence of Ca(2+) influx, buffers influence the temporal characteristics, but not the magnitude, of [Ca(2+)] fluctuations. We derive an analytical formula describing the influence of rapid Ca(2+) buffers on [Ca(2+)] fluctuations and, importantly, identify the stochastic analog of (deterministic) effective domain volume. Our results demonstrate that Ca(2+) buffers alter the dynamics of [Ca(2+)] fluctuations in a nonintuitive manner. The finding that Ca(2

  10. Topographic specificity of functional connections from hippocampal CA3 to CA1

    NASA Astrophysics Data System (ADS)

    Brivanlou, Iman H.; Dantzker, Jami L. M.; Stevens, Charles F.; Callaway, Edward M.

    2004-02-01

    The hippocampus is a cortical region thought to play an important role in learning and memory. Most of our knowledge about the detailed organization of hippocampal circuitry responsible for these functions is derived from anatomical studies. These studies present an incomplete picture, however, because the functional character and importance of connections are often not revealed by anatomy. Here, we used a physiological method (photostimulation with caged glutamate) to probe the fine pattern of functional connectivity between the CA3 and CA1 subfields in the mouse hippocampal slice preparation. We recorded intracellularly from CA1 and CA3 pyramidal neurons while scanning with photostimulation across the entire CA3 subfield with high spatial resolution. Our results show that, at a given septotemporal level, nearby CA1 neurons receive synaptic inputs from neighboring CA3 neurons. Thus, the CA3 to CA1 mapping preserves neighbor relations.

  11. Dietary calcium deficiency increases Ca2+ uptake and Ca2+ extrusion mechanisms in chick enterocytes.

    PubMed

    Centeno, Viviana A; Díaz de Barboza, Gabriela E; Marchionatti, Ana M; Alisio, Arturo E; Dallorso, Maria E; Nasif, Renée; Tolosa de Talamoni, Nori G

    2004-10-01

    Ca2+ uptake and Ca2+ extrusion mechanisms were studied in enterocytes with different degree of differentiation from chicks adapted to a low Ca2+ diet as compared to animals fed a normal diet. Chicks adapted to a low Ca2+ diet presented hypocalcemia, hypophosphatemia and increased serum 1,25(OH)2D3 and Ca2+ absorption. Low Ca2+ diet increased the alkaline phosphatase (AP) activity, independently of the cellular maturation, but it did not alter gamma-glutamyl-transpeptidase activity. Ca2+ uptake, Ca2+-ATPase and Na(+)/Ca2+ exchanger activities and expressions were increased by the mineral-deficient diet either in mature or immature enterocytes. Western blots analysis shows that vitamin D receptor (VDR) expression was much higher in crypt cells than in mature cells. Low Ca2+ diet decreased the number of vitamin D receptor units in both kinds of cells. In conclusion, changes in Ca2+ uptake and Ca2+ extrusion mechanisms in the enterocytes by a low Ca2+ diet appear to be a result of enhanced serum levels of 1,25(OH)2D3, which would promote cellular differentiation producing cells more efficient to express vitamin D dependent genes required for Ca2+ absorption. PMID:15528161

  12. TMCO1 Is an ER Ca(2+) Load-Activated Ca(2+) Channel.

    PubMed

    Wang, Qiao-Chu; Zheng, Qiaoxia; Tan, Haiyan; Zhang, Bing; Li, Xiaoling; Yang, Yuxiu; Yu, Jie; Liu, Yang; Chai, Hao; Wang, Xi; Sun, Zhongshuai; Wang, Jiu-Qiang; Zhu, Shu; Wang, Fengli; Yang, Maojun; Guo, Caixia; Wang, Heng; Zheng, Qingyin; Li, Yang; Chen, Quan; Zhou, Aimin; Tang, Tie-Shan

    2016-06-01

    Maintaining homeostasis of Ca(2+) stores in the endoplasmic reticulum (ER) is crucial for proper Ca(2+) signaling and key cellular functions. The Ca(2+)-release-activated Ca(2+) (CRAC) channel is responsible for Ca(2+) influx and refilling after store depletion, but how cells cope with excess Ca(2+) when ER stores are overloaded is unclear. We show that TMCO1 is an ER transmembrane protein that actively prevents Ca(2+) stores from overfilling, acting as what we term a "Ca(2+) load-activated Ca(2+) channel" or "CLAC" channel. TMCO1 undergoes reversible homotetramerization in response to ER Ca(2+) overloading and disassembly upon Ca(2+) depletion and forms a Ca(2+)-selective ion channel on giant liposomes. TMCO1 knockout mice reproduce the main clinical features of human cerebrofaciothoracic (CFT) dysplasia spectrum, a developmental disorder linked to TMCO1 dysfunction, and exhibit severe mishandling of ER Ca(2+) in cells. Our findings indicate that TMCO1 provides a protective mechanism to prevent overfilling of ER stores with Ca(2+) ions. PMID:27212239

  13. Ca2+ cycling in heart cells from ground squirrels: adaptive strategies for intracellular Ca2+ homeostasis.

    PubMed

    Li, Xiao-Chen; Wei, Ling; Zhang, Guang-Qin; Bai, Zai-Ling; Hu, Ying-Ying; Zhou, Peng; Bai, Shu-Hua; Chai, Zhen; Lakatta, Edward G; Hao, Xue-Mei; Wang, Shi-Qiang

    2011-01-01

    Heart tissues from hibernating mammals, such as ground squirrels, are able to endure hypothermia, hypoxia and other extreme insulting factors that are fatal for human and nonhibernating mammals. This study was designed to understand adaptive mechanisms involved in intracellular Ca(2+) homeostasis in cardiomyocytes from the mammalian hibernator, ground squirrel, compared to rat. Electrophysiological and confocal imaging experiments showed that the voltage-dependence of L-type Ca(2+) current (I(Ca)) was shifted to higher potentials in ventricular myocytes from ground squirrels vs. rats. The elevated threshold of I(Ca) did not compromise the Ca(2+)-induced Ca(2+) release, because a higher depolarization rate and a longer duration of action potential compensated the voltage shift of I(Ca). Both the caffeine-sensitive and caffeine-resistant components of cytosolic Ca(2+) removal were more rapid in ground squirrels. Ca(2+) sparks in ground squirrels exhibited larger amplitude/size and much lower frequency than in rats. Due to the high I(Ca) threshold, low SR Ca(2+) leak and rapid cytosolic Ca(2+) clearance, heart cells from ground squirrels exhibited better capability in maintaining intracellular Ca(2+) homeostasis than those from rats and other nonhibernating mammals. These findings not only reveal adaptive mechanisms of hibernation, but also provide novel strategies against Ca(2+) overload-related heart diseases. PMID:21935466

  14. Anodic reactions in the Ca/CaCrO/sub 4/ thermal battery

    SciTech Connect

    Guidotti, R.A.; Reinhardt, F.W.

    1985-09-01

    The reaction of Ca with a CaCrO/sub 4/-(LiCl-KCl eutectic) solution at temperatures of 400/sup 0/C to 500/sup 0/C was studied to better understand the nature of the chemical reactions and electrochemical processes that occur in the Ca/CaCrO/sub 4/ thermal battery at the anode during activation and discharge. Limited tests also were conducted with a CaCrO/sub 4/-(CaCl/sub 2/-NaCl-KCl eutectic) solution at 550/sup 0/C. Ca/CaCrO/sub 4/ and CaLi/sub 2//CaCrO/sub 4/ single cells were tested to observe the relative performance differences of Ca and CaLi/sub 2/ anodes. The discharged cells were analyzed by optical microscopy, electron microprobe, Auger electron spectroscopy, and secondary-ion mass spectroscopy. These analytical data were used in conjunction with the results of chemical-reaction experiments to propose a discharge mechanism for the Ca/CaCrO/sub 4/ thermal battery, consistent with experimental observations.

  15. Anodic reactions in the Ca/CaCrO4 thermal battery

    NASA Astrophysics Data System (ADS)

    Guidotti, R. A.; Reinhardt, F. W.

    1985-09-01

    The reaction of Ca with a CaCrO4-(LiCl-KCl eutectic) solution at temperatures of 400(0)C to 500(0)C was studied to better understand the nature of the chemical reactions and electrochemical processes that occur in the Ca/CaCrO4 thermal battery at the anode during activation and discharge. Limited tests also were conducted with a CaCrO4-(CaCl2-NaCl-KCl eutectic) solution at 550(0)C. Ca/CaCrO4 and CaLi2/CaCrO4 single cells were tested to observe the relative performance differences of Ca and CaLi2 anodes. The discharged cells were analyzed by optical microscopy, electron microprobe, Auger electron spectroscopy, and secondary-ion mass spectroscopy. These analytical data were used in conjunction with the results of chemical-reaction experiments to propose a discharge mechanism for the Ca/CaCrO4 thermal battery, consistent with experimental observations.

  16. Serum CA549 in primary breast cancer: comparison with CA15.3 and MCA.

    PubMed

    Gion, M; Plebani, M; Mione, R; Penzo, C; Meo, S; Burlina, A

    1994-04-01

    We carried out a comparison of three commonly used mucin markers, CA549, CA15.3 and MCA. Serum samples from 184 healthy women and 237 patients with primary breast cancer were evaluated. The markers were measured using commercially available immunometric assays. Like CA15.3 and MCA, CA549 was significantly associated with tumour size and lymph node status, being an effective indicator of tumour bulk. CA549 was significantly correlated with both CA15.3 and MCA. Positive/negative concordance rate was very good (93.7%) between CA549 and MCA. Conversely, CA15.3 was positive and CA549 negative in 20.4% of cases. Axillary status was not significantly different in the latter group of patients and in cases in which CA15.3 and CA549 showed concordant results. From the present findings we draw the following major conclusions: 1. CA549 and MCA are highly correlated and their association should not provide additional information; however, they should not be considered interchangeable since they may behave differently in individual cases. 2. CA549 and CA15.3, although well correlated, are discordant in a significant number of cases. Longitudinal studies are needed to verify the usefulness of the association between the two markers. 3. The three evaluated mucin markers are not interchangeable in individual patients; if a patient is monitored with a marker, she should be followed up with the same marker.

  17. Dissection of local Ca(2+) signals inside cytosol by ER-targeted Ca(2+) indicator.

    PubMed

    Niwa, Fumihiro; Sakuragi, Shigeo; Kobayashi, Ayana; Takagi, Shin; Oda, Yoichi; Bannai, Hiroko; Mikoshiba, Katsuhiko

    2016-10-01

    Calcium (Ca(2+)) is a versatile intracellular second messenger that operates in various signaling pathways leading to multiple biological outputs. The diversity of spatiotemporal patterns of Ca(2+) signals, generated by the coordination of Ca(2+) influx from the extracellular space and Ca(2+) release from the intracellular Ca(2+) store the endoplasmic reticulum (ER), is considered to underlie the diversity of biological outputs caused by a single signaling molecule. However, such Ca(2+) signaling diversity has not been well described because of technical limitations. Here, we describe a new method to report Ca(2+) signals at subcellular resolution. We report that OER-GCaMP6f, a genetically encoded Ca(2+) indicator (GECI) targeted to the outer ER membrane, can monitor Ca(2+) release from the ER at higher spatiotemporal resolution than conventional GCaMP6f. OER-GCaMP6f was used for in vivo Ca(2+) imaging of C. elegans. We also found that the spontaneous Ca(2+) elevation in cultured astrocytes reported by OER-GCaMP6f showed a distinct spatiotemporal pattern from that monitored by plasma membrane-targeted GCaMP6f (Lck-GCaMP6f); less frequent Ca(2+) signal was detected by OER-GCaMP6f, in spite of the fact that Ca(2+) release from the ER plays important roles in astrocytes. These findings suggest that targeting of GECIs to the ER outer membrane enables sensitive detection of Ca(2+) release from the ER at subcellular resolution, avoiding the diffusion of GECI and Ca(2+). Our results indicate that Ca(2+) imaging with OER-GCaMP6f in combination with Lck-GCaMP6f can contribute to describing the diversity of Ca(2+) signals, by enabling dissection of Ca(2+) signals at subcellular resolution.

  18. Transcriptional Mechanisms Regulating Ca2+ Homeostasis

    PubMed Central

    Ritchie, Michael F.; Zhou, Yandong; Soboloff, Jonathan

    2011-01-01

    Ca2+ is a dynamic cellular secondary messenger which mediates a vast array of cellular responses. Control over these processes is achieved via an extensive combination of pumps and channels which regulate the concentration of Ca2+ within not only the cytosol but also all intracellular compartments. Precisely how these pumps and channels are regulated is only partially understood, however, recent investigations have identified members of the Early Growth Response (EGR) family of zinc finger transcription factors as critical players in this process. The roles of several other transcription factors in control of Ca2+ homeostasis have also been demonstrated, including Wilms Tumor Suppressor 1 (WT1), Nuclear Factor of Activated T cells (NFAT) and c-myc. In this review, we will discuss not only how these transcription factors regulate the expression of the major proteins involved in control of Ca2+ homeostasis, but also how this transcriptional remodeling of Ca2+ homeostasis affects Ca2+ dynamics and cellular responses. PMID:21074851

  19. Large Ca isotope effect in the CaC{sub 6} superconductor.

    SciTech Connect

    Hinks, D. G.; Rosenmann, D.; Claus, H.; Bailey, M. S.; Jorgensen, J. D.; Materials Science Division

    2007-01-01

    We have measured the Ca isotope effect coefficient, {alpha}(Ca), in the newly discovered superconductor CaC{sub 6} and find a value of 0.53(2). This result shows that the superconductivity is dominated by coupling of the electrons by Ca phonon modes. The C phonons contribute very little, assuming that this material is a conventional electron-phonon coupled superconductor. Thus, in contrast to another layered material MgB{sub 2}, where high-energy phonons in the B layers are responsible for the superconductivity, in layered CaC{sub 6} the phonons responsible for superconductivity are primarily low-energy modes of the intercalated Ca.

  20. Glutamate excitotoxicity and Ca2+-regulation of respiration: Role of the Ca2+ activated mitochondrial transporters (CaMCs).

    PubMed

    Rueda, Carlos B; Llorente-Folch, Irene; Traba, Javier; Amigo, Ignacio; Gonzalez-Sanchez, Paloma; Contreras, Laura; Juaristi, Inés; Martinez-Valero, Paula; Pardo, Beatriz; Del Arco, Araceli; Satrustegui, Jorgina

    2016-08-01

    Glutamate elicits Ca(2+) signals and workloads that regulate neuronal fate both in physiological and pathological circumstances. Oxidative phosphorylation is required in order to respond to the metabolic challenge caused by glutamate. In response to physiological glutamate signals, cytosolic Ca(2+) activates respiration by stimulation of the NADH malate-aspartate shuttle through Ca(2+)-binding to the mitochondrial aspartate/glutamate carrier (Aralar/AGC1/Slc25a12), and by stimulation of adenine nucleotide uptake through Ca(2+) binding to the mitochondrial ATP-Mg/Pi carrier (SCaMC-3/Slc25a23). In addition, after Ca(2+) entry into the matrix through the mitochondrial Ca(2+) uniporter (MCU), it activates mitochondrial dehydrogenases. In response to pathological glutamate stimulation during excitotoxicity, Ca(2+) overload, reactive oxygen species (ROS), mitochondrial dysfunction and delayed Ca(2+) deregulation (DCD) lead to neuronal death. Glutamate-induced respiratory stimulation is rapidly inactivated through a mechanism involving Poly (ADP-ribose) Polymerase-1 (PARP-1) activation, consumption of cytosolic NAD(+), a decrease in matrix ATP and restricted substrate supply. Glutamate-induced Ca(2+)-activation of SCaMC-3 imports adenine nucleotides into mitochondria, counteracting the depletion of matrix ATP and the impaired respiration, while Aralar-dependent lactate metabolism prevents substrate exhaustion. A second mechanism induced by excitotoxic glutamate is permeability transition pore (PTP) opening, which critically depends on ROS production and matrix Ca(2+) entry through the MCU. By increasing matrix content of adenine nucleotides, SCaMC-3 activity protects against glutamate-induced PTP opening and lowers matrix free Ca(2+), resulting in protracted appearance of DCD and protection against excitotoxicity in vitro and in vivo, while the lack of lactate protection during in vivo excitotoxicity explains increased vulnerability to kainite-induced toxicity in Aralar

  1. Ca2+ signaling differentiation during oocyte maturation.

    PubMed

    Machaca, Khaled

    2007-11-01

    Oocyte maturation is an essential cellular differentiation pathway that prepares the egg for activation at fertilization leading to the initiation of embryogenesis. An integral attribute of oocyte maturation is the remodeling of Ca2+ signaling pathways endowing the egg with the capacity to produce a specialized Ca2+ transient at fertilization that is necessary and sufficient for egg activation. Consequently, mechanistic elucidation of Ca2+ signaling differentiation during oocyte maturation is fundamental to our understanding of egg activation, and offers a glimpse into Ca2+ signaling regulation during the cell cycle.

  2. Genetically Encoded Green Fluorescent Ca2+ Indicators with Improved Detectability for Neuronal Ca2+ Signals

    PubMed Central

    Sadakari, Junko; Gengyo-Ando, Keiko; Kagawa-Nagamura, Yuko; Kobayashi, Chiaki; Ikegaya, Yuji; Nakai, Junichi

    2012-01-01

    Imaging the activities of individual neurons with genetically encoded Ca2+ indicators (GECIs) is a promising method for understanding neuronal network functions. Here, we report GECIs with improved neuronal Ca2+ signal detectability, termed G-CaMP6 and G-CaMP8. Compared to a series of existing G-CaMPs, G-CaMP6 showed fairly high sensitivity and rapid kinetics, both of which are suitable properties for detecting subtle and fast neuronal activities. G-CaMP8 showed a greater signal (Fmax/Fmin = 38) than G-CaMP6 and demonstrated kinetics similar to those of G-CaMP6. Both GECIs could detect individual spikes from pyramidal neurons of cultured hippocampal slices or acute cortical slices with 100% detection rates, demonstrating their superior performance to existing GECIs. Because G-CaMP6 showed a higher sensitivity and brighter baseline fluorescence than G-CaMP8 in a cellular environment, we applied G-CaMP6 for Ca2+ imaging of dendritic spines, the putative postsynaptic sites. By expressing a G-CaMP6-actin fusion protein for the spines in hippocampal CA3 pyramidal neurons and electrically stimulating the granule cells of the dentate gyrus, which innervate CA3 pyramidal neurons, we found that sub-threshold stimulation triggered small Ca2+ responses in a limited number of spines with a low response rate in active spines, whereas supra-threshold stimulation triggered large fluorescence responses in virtually all of the spines with a 100% activity rate. PMID:23240011

  3. Extrapolating microdomain Ca2+ dynamics using BK channels as a Ca2+ sensor

    PubMed Central

    Hou, Panpan; Xiao, Feng; Liu, Haowen; Yuchi, Ming; Zhang, Guohui; Wu, Ying; Wang, Wei; Zeng, Wenping; Ding, Mingyue; Cui, Jianming; Wu, Zhengxing; Wang, Lu-Yang; Ding, Jiuping

    2016-01-01

    Ca2+ ions play crucial roles in mediating physiological and pathophysiological processes, yet Ca2+ dynamics local to the Ca2+ source, either from influx via calcium permeable ion channels on plasmic membrane or release from internal Ca2+ stores, is difficult to delineate. Large-conductance calcium-activated K+ (BK-type) channels, abundantly distribute in excitable cells and often localize to the proximity of voltage-gated Ca2+ channels (VGCCs), spatially enabling the coupling of the intracellular Ca2+ signal to the channel gating to regulate membrane excitability and spike firing patterns. Here we utilized the sensitivity and dynamic range of BK to explore non-uniform Ca2+ local transients in the microdomain of VGCCs. Accordingly, we applied flash photolysis of caged Ca2+ to activate BK channels and determine their intrinsic sensitivity to Ca2+. We found that uncaging Ca2+ activated biphasic BK currents with fast and slow components (time constants being τf ≈ 0.2 ms and τs ≈ 10 ms), which can be accounted for by biphasic Ca2+ transients following light photolysis. We estimated the Ca2+-binding rate constant kb (≈1.8 × 108 M−1s−1) for mSlo1 and further developed a model in which BK channels act as a calcium sensor capable of quantitatively predicting local microdomain Ca2+ transients in the vicinity of VGCCs during action potentials. PMID:26776352

  4. Distinct roles for dorsal CA3 and CA1 in memory for sequential nonspatial events.

    PubMed

    Farovik, Anja; Dupont, Laura M; Eichenbaum, Howard

    2010-01-01

    Previous studies have suggested that dorsal hippocampal areas CA3 and CA1 are both involved in representing sequences of events that compose unique episodes. However, it is uncertain whether the contribution of CA3 is restricted to spatial information, and it is unclear whether CA1 encodes order per se or contributes by an active maintenance of memories of sequential events. Here, we developed a new behavioral task that examines memory for the order of sequential nonspatial events presented as trial-unique odor pairings. When the interval between odors within a studied pair was brief (3 sec), bilateral dorsal CA3 lesions severely disrupted memory for their order, whereas dorsal CA1 lesions did not affect performance. However, when the inter-item interval was extended to 10 sec, CA1 lesions, as well as CA3 lesions, severely disrupted performance. These findings suggest that the role of CA3 in sequence memory is not limited to spatial information, but rather appears to be a fundamental property of CA3 function. In contrast, CA1 becomes involved when memories for events must be held or sequenced over long intervals. Thus, CA3 and CA1 are both involved in memory for sequential nonspatial events that compose unique experiences, and these areas play different roles that are distinguished by the duration of time that must be bridged between key events.

  5. Conservation of Ca2+/Calmodulin Regulation across Na and Ca2+ channels

    PubMed Central

    Ben-Johny, Manu; Yang, Philemon S.; Niu, Jacqueline; Yang, Wanjun; Joshi-Mukherjee, Rosy; Yue, David T.

    2014-01-01

    SUMMARY Voltage-gated Na and Ca2+channels comprise distinct ion-channel superfamilies, yet the carboxy tails of these channels exhibit high homology hinting at a long-shared and purposeful module. For different Ca2+ channels, carboxyl-tail inter actions with calmodulin do elaborate robust and similar forms of Ca2+ regulation. However, Na channels have only shown subtler Ca2+modulation that differs among reports, challenging attempts at unified understanding. Here, by rapid Ca2+photoreleaseon to Na channels, we reset this view of Na channel regulation. For cardiac muscle channels (NaV1.5), reported effects from which most mechanistic proposals derive, we observe no Ca2+modulation. Conversely, for skeletal-muscle channels (NaV1.4), we uncover fast Ca2+ regulation eerily similar to that of Ca2+ channels. Channel opathic myotonia mutations halve NaV1.4 Ca2+ regulation, and transplanting the NaV1.4 carboxy tail onto Ca2+ channels recapitulates Ca2+ regulation. Thus we argue for the persistence and physiological relevance of an ancient Ca2+ regulatory module across Na and Ca2+ channels. PMID:24949975

  6. Biphasic decay of the Ca transient results from increased sarcoplasmic reticulum Ca leak

    PubMed Central

    Sankaranarayanan, Rajiv; Li, Yatong; Greensmith, David J.; Eisner, David A.

    2016-01-01

    Key points Ca leak from the sarcoplasmic reticulum through the ryanodine receptor (RyR) reduces the amplitude of the Ca transient and slows its rate of decay.In the presence of β‐adrenergic stimulation, RyR‐mediated Ca leak produces a biphasic decay of the Ca transient with a fast early phase and a slow late phase.Two forms of Ca leak have been studied, Ca‐sensitising (induced by caffeine) and non‐sensitising (induced by ryanodine) and both induce biphasic decay of the Ca transient.Only Ca‐sensitising leak can be reversed by traditional RyR inhibitors such as tetracaine.Ca leak can also induce Ca waves. At low levels of leak, waves occur. As leak is increased, first biphasic decay and then slowed monophasic decay is seen. The level of leak has major effects on the shape of the Ca transient. Abstract In heart failure, a reduction in Ca transient amplitude and contractile dysfunction can by caused by Ca leak through the sarcoplasmic reticulum (SR) Ca channel (ryanodine receptor, RyR) and/or decreased activity of the SR Ca ATPase (SERCA). We have characterised the effects of two forms of Ca leak (Ca‐sensitising and non‐sensitising) on calcium cycling and compared with those of SERCA inhibition. We measured [Ca2+]i with fluo‐3 in voltage‐clamped rat ventricular myocytes. Increasing SR leak with either caffeine (to sensitise the RyR to Ca activation) or ryanodine (non‐sensitising) had similar effects to SERCA inhibition: decreased systolic [Ca2+]i, increased diastolic [Ca2+]i and slowed decay. However, in the presence of isoproterenol, leak produced a biphasic decay of the Ca transient in the majority of cells while SERCA inhibition produced monophasic decay. Tetracaine reversed the effects of caffeine but not of ryanodine. When caffeine (1 mmol l−1) was added to a cell which displayed Ca waves, the wave frequency initially increased before waves disappeared and biphasic decay developed. Eventually (at higher caffeine concentrations), the

  7. HISTORIC IMAGE: VIEW OF CA. 1890SERA ROSTRUM, DEMOLISHED. PHOTOGRAPH CA. ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    HISTORIC IMAGE: VIEW OF CA. 1890S-ERA ROSTRUM, DEMOLISHED. PHOTOGRAPH CA. 1930S. CEMETERY MAINTENANCE LEDGER, NCA HISTORY COLLECTION. - Alexandria National Cemetery, 1450 Wilkes Street, Alexandria, Independent City, VA

  8. Submicroscopic Ca2+ diffusion mediates inhibitory coupling between individual Ca2+ channels.

    PubMed

    Imredy, J P; Yue, D T

    1992-08-01

    Dihydropyridine-sensitive Ca2+ channels in heart demonstrate an important negative feedback property: they close, or inactivate, in response to prior Ca2+ entry. We now find that Ca2+ influx through one channel can selectively contribute to the inactivation of another adjacent channel, without a generalized elevation of bulk intracellular Ca2+ concentration. Intracellular application of the Ca2+ chelator BAPTA greatly diminishes such negative interactions within Ca2+ channel pairs. These findings demonstrate that Ca2+ currents are controlled not only by intrinsic channel properties, but also by local diffusive interactions among neighboring channels. Such inhibitory coupling among channels provides a concrete example of localized Ca2+ signaling, long proposed to exist on the basis of theoretical calculations. PMID:1323309

  9. By Regulating Mitochondrial Ca2+-Uptake UCP2 Modulates Intracellular Ca2+

    PubMed Central

    Gebing, Tina; Reda, Sara; Schwaiger, Astrid; Leitner, Johannes; Wolny, Martin; Eckardt, Lars; Hoppe, Uta C.

    2016-01-01

    Introduction The possible role of UCP2 in modulating mitochondrial Ca2+-uptake (mCa2+-uptake) via the mitochondrial calcium uniporter (MCU) is highly controversial. Methods Thus, we analyzed mCa2+-uptake in isolated cardiac mitochondria, MCU single-channel activity in cardiac mitoplasts, dual Ca2+-transients from mitochondrial ((Ca2+)m) and intracellular compartment ((Ca2+)c) in the whole-cell configuration in cardiomyocytes of wild-type (WT) and UCP2-/- mice. Results Isolated mitochondria showed a Ru360 sensitive mCa2+-uptake, which was significantly decreased in UCP2-/- (229.4±30.8 FU vs. 146.3±23.4 FU, P<0.05). Single-channel registrations confirmed a Ru360 sensitive voltage-gated Ca2+-channel in mitoplasts, i.e. mCa1, showing a reduced single-channel activity in UCP2-/- (Po,total: 0.34±0.05% vs. 0.07±0.01%, P<0.05). In UCP2-/- cardiomyocytes (Ca2+)m was decreased (0.050±0.009 FU vs. 0.021±0.005 FU, P<0.05) while (Ca2+)c was unchanged (0.032±0.002 FU vs. 0.028±0.004 FU, P>0.05) and transsarcolemmal Ca2+-influx was inhibited suggesting a possible compensatory mechanism. Additionally, we observed an inhibitory effect of ATP on mCa2+-uptake in WT mitoplasts and (Ca2+)m of cardiomyocytes leading to an increase of (Ca2+)c while no ATP dependent effect was observed in UCP2-/-. Conclusion Our results indicate regulatory effects of UCP2 on mCa2+-uptake. Furthermore, we propose, that previously described inhibitory effects on MCU by ATP may be mediated via UCP2 resulting in changes of excitation contraction coupling. PMID:26849136

  10. 46 CFR 7.130 - Point Conception, CA to Point Sur, CA.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 1 2010-10-01 2010-10-01 false Point Conception, CA to Point Sur, CA. 7.130 Section 7.130 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY PROCEDURES APPLICABLE TO THE PUBLIC BOUNDARY LINES Pacific Coast § 7.130 Point Conception, CA to Point Sur, CA. (a) A line drawn from the southernmost extremity of Fossil Point at longitude...

  11. 46 CFR 7.125 - Point Vincente, CA to Point Conception, CA.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 1 2010-10-01 2010-10-01 false Point Vincente, CA to Point Conception, CA. 7.125 Section 7.125 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY PROCEDURES APPLICABLE TO THE PUBLIC BOUNDARY LINES Pacific Coast § 7.125 Point Vincente, CA to Point Conception, CA. (a) A line drawn from Redondo Beach East Jetty Light “2” to...

  12. Effect of Ca2+ efflux pathway distribution and exogenous Ca2+ buffers on intracellular Ca2+ dynamics in the rat ventricular myocyte: a simulation study.

    PubMed

    Pásek, Michal; Simurda, Jiří; Orchard, Clive H

    2014-01-01

    We have used a previously published computer model of the rat cardiac ventricular myocyte to investigate the effect of changing the distribution of Ca(2+) efflux pathways (SERCA, Na(+)/Ca(2+) exchange, and sarcolemmal Ca(2+) ATPase) between the dyad and bulk cytoplasm and the effect of adding exogenous Ca(2+) buffers (BAPTA or EGTA), which are used experimentally to differentially buffer Ca(2+) in the dyad and bulk cytoplasm, on cellular Ca(2+) cycling. Increasing the dyadic fraction of a particular Ca(2+) efflux pathway increases the amount of Ca(2+) removed by that pathway, with corresponding changes in Ca(2+) efflux from the bulk cytoplasm. The magnitude of these effects varies with the proportion of the total Ca(2+) removed from the cytoplasm by that pathway. Differences in the response to EGTA and BAPTA, including changes in Ca(2+)-dependent inactivation of the L-type Ca(2+) current, resulted from the buffers acting as slow and fast "shuttles," respectively, removing Ca(2+) from the dyadic space. The data suggest that complex changes in dyadic Ca(2+) and cellular Ca(2+) cycling occur as a result of changes in the location of Ca(2+) removal pathways or the presence of exogenous Ca(2+) buffers, although changing the distribution of Ca(2+) efflux pathways has relatively small effects on the systolic Ca(2+) transient.

  13. Literacy.CA. Issue #19, Winter 2006

    ERIC Educational Resources Information Center

    Murray, Fiona, Ed.

    2006-01-01

    The "literacy.ca" newsletter is a vehicle for literacy workers and supporters to share information, ideas, resources and research on emerging literacy issues. This issue of "literacy.ca" contains the following articles: (1) Riding The Wave: How will the federal election affect progress on a pan-Canadian literacy agenda?; (2) Point of View:…

  14. Causes-of-death analysis of patients with cardiac resynchronization therapy: an analysis of the CeRtiTuDe cohort study

    PubMed Central

    Marijon, Eloi; Leclercq, Christophe; Narayanan, Kumar; Boveda, Serge; Klug, Didier; Lacaze-Gadonneix, Jonathan; Defaye, Pascal; Jacob, Sophie; Piot, Olivier; Deharo, Jean-Claude; Perier, Marie-Cecile; Mulak, Genevieve; Hermida, Jean-Sylvain; Milliez, Paul; Gras, Daniel; Cesari, Olivier; Hidden-Lucet, Françoise; Anselme, Frederic; Chevalier, Philippe; Maury, Philippe; Sadoul, Nicolas; Bordachar, Pierre; Cazeau, Serge; Chauvin, Michel; Empana, Jean-Philippe; Jouven, Xavier; Daubert, Jean-Claude; Le Heuzey, Jean-Yves

    2015-01-01

    Aims The choice of resynchronization therapy between with (CRT-D) and without (CRT-P) a defibrillator remains a contentious issue. Cause-of-death analysis among CRT-P, compared with CRT-D, patients could help evaluate the extent to which CRT-P patients would have additionally benefited from a defibrillator in a daily clinical practice. Methods and results A total of 1705 consecutive patients implanted with a CRT (CRT-P: 535 and CRT-D: 1170) between 2008 and 2010 were enrolled in CeRtiTuDe, a multicentric prospective follow-up cohort study, with specific adjudication for causes of death at 2 years. Patients with CRT-P compared with CRT-D were older (P < 0.0001), less often male (P < 0.0001), more symptomatic (P = 0.0005), with less coronary artery disease (P = 0.003), wider QRS (P = 0.002), more atrial fibrillation (P < 0.0001), and more co-morbidities (P = 0.04). At 2-year follow-up, the annual overall mortality rate was 83.80 [95% confidence interval (CI) 73.41–94.19] per 1000 person-years. The crude mortality rate among CRT-P patients was double compared with CRT-D (relative risk 2.01, 95% CI 1.56–2.58). In a Cox proportional hazards regression analysis, CRT-P remained associated with increased mortality (hazard ratio 1.54, 95% CI 1.07–2.21, P = 0.0209), although other potential confounders may persist. By cause-of-death analysis, 95% of the excess mortality among CRT-P subjects was related to an increase in non-sudden death. Conclusion When compared with CRT-D patients, excess mortality in CRT-P recipients was mainly due to non-sudden death. Our findings suggest that CRT-P patients, as currently selected in routine clinical practice, would not potentially benefit with the addition of a defibrillator. PMID:26330420

  15. CA-MRSA. The new sports pathogen.

    PubMed

    Kurkowski, Christina

    2007-01-01

    Skin infections in athletes caused by community-associated Methicillin-resistant Staphylococcus aureus (CA-MRSA) have been observed within many cities throughout the United States and within many countries throughout the world (Centers for Disease Control and Prevention [CDC], 2003). As the incidence rises in the athletic population, clinicians must learn to identify risk factors for CA-MRSA, diagnosis and treat infections with judicious use of antimicrobial agents and facilitate strategies to limit transmission. Recently, a new consensus guideline for handling CA-MRSA outbreaks in sports has been released by the CDC (Gorwitz et al., 2006). This article includes a review of the evolution of MRSA; distinguishes between healthcare associated Methicillin-resistant Staphylococcus aureus (HA-MRSA) and CA-MRSA; and reviews the diagnosis, management, and prevention strategies to limit transmission of CA-MRSA.

  16. Ca-Dependent Folding of Human Calumenin.

    PubMed

    Mazzorana, Marco; Hussain, Rohanah; Sorensen, Thomas

    2016-01-01

    Human calumenin (hCALU) is a six EF-hand protein belonging to the CREC family. As other members of the family, it is localized in the secretory pathway and regulates the activity of SERCA2a and of the ryanodine receptor in the endoplasmic reticulum (ER). We have studied the effects of Ca2+ binding to the protein and found it to attain a more compact structure upon ion binding. Circular Dichroism (CD) measurements suggest a major rearrangement of the protein secondary structure, which reversibly switches from disordered at low Ca2+ concentrations to predominantly alpha-helical when Ca2+ is added. SAXS experiments confirm the transition from an unfolded to a compact structure, which matches the structural prediction of a trilobal fold. Overall our experiments suggest that calumenin is a Ca2+ sensor, which folds into a compact structure, capable of interacting with its molecular partners, when Ca2+ concentration within the ER reaches the millimolar range. PMID:26991433

  17. Vasospastic angina and Ca channel blockers.

    PubMed

    Minatoguchi, Shinya

    2013-08-01

    Coronary artery spasm is one of the causes of angina pectoris,acute myocardial infarction and ventricular fibrillation-related sudden death. It has been established that Ca channel blockers are protective against vasospastic angina (VSA) and treatment with Ca channel blockers provides a better prognosis of VSA. However, it is not still clarified what kinds of Ca channel blockers shows the best prognosis of VSA. We performed a meta-analysis in which 4Ca channel blockers amlodipine, nifedipine, benidipine and diltiazem were used for the treatment of VSA patients and found that among 4 Ca channel blockers, benidipine showed a statistically significant better prognostic effect on MACE than amlodipine, nifedipine or diltiazem.

  18. Ca-Dependent Folding of Human Calumenin

    PubMed Central

    Mazzorana, Marco; Hussain, Rohanah; Sorensen, Thomas

    2016-01-01

    Human calumenin (hCALU) is a six EF-hand protein belonging to the CREC family. As other members of the family, it is localized in the secretory pathway and regulates the activity of SERCA2a and of the ryanodine receptor in the endoplasmic reticulum (ER). We have studied the effects of Ca2+ binding to the protein and found it to attain a more compact structure upon ion binding. Circular Dichroism (CD) measurements suggest a major rearrangement of the protein secondary structure, which reversibly switches from disordered at low Ca2+ concentrations to predominantly alpha-helical when Ca2+ is added. SAXS experiments confirm the transition from an unfolded to a compact structure, which matches the structural prediction of a trilobal fold. Overall our experiments suggest that calumenin is a Ca2+ sensor, which folds into a compact structure, capable of interacting with its molecular partners, when Ca2+ concentration within the ER reaches the millimolar range. PMID:26991433

  19. The CaMKK2/CaMKIV Relay Is an Essential Regulator of Hepatic Cancer

    PubMed Central

    Lin, Fumin; Marcelo, Kathrina L.; Rajapakshe, Kimal; Coarfa, Cristian; Dean, Adam; Wilganowski, Nathaniel; Robinson, Holly; Sevick, Eva; Bissig, Karl-Dimiter; Goldie, Lauren C.; Means, Anthony R.; York, Brian

    2015-01-01

    Hepatic cancer is one of the most lethal cancers worldwide. Here, we report that the expression of Ca2+/calmodulin-dependent protein kinase kinase 2 (CaMKK2) is significantly up-regulated in hepatocellular carcinoma (HCC) and negatively correlated with HCC patient survival. The CaMKK2 protein is highly expressed in all eight hepatic cancer cell lines evaluated and is markedly up-regulated relative to normal primary hepatocytes. Loss of CaMKK2 function is sufficient to inhibit liver cancer cell growth, and the growth defect resulting from loss of CaMKK2 can be rescued by ectopic expression of wild-type CaMKK2 but not by kinase-inactive mutants. Cellular ablation of CaMKK2 using RNA interference yields a gene signature that correlates with improvement in HCC patient survival, and ablation or pharmacological inhibition of CaMKK2 with STO-609 impairs tumorigenicity of liver cancer cells in vivo. Moreover, CaMKK2 expression is up-regulated in a time-dependent manner in a carcinogen-induced HCC mouse model, and STO-609 treatment regresses hepatic tumor burden in this model. Mechanistically, CaMKK2 signals through Ca2+/calmodulin-dependent protein kinase 4 (CaMKIV) to control liver cancer cell growth. Further analysis revealed that CaMKK2 serves as a scaffold to assemble CaMKIV with key components of the mammalian target of rapamycin/ribosomal protein S6 kinase, 70 kDa, pathway and thereby stimulate protein synthesis through protein phosphorylation. Conclusion The CaMKK2/CaMKIV relay is an upstream regulator of the oncogenic mammalian target of rapamycin/ribosomal protein S6 kinase, 70 kDa, pathway, and the importance of this CaMKK2/CaM-KIV axis in HCC growth is confirmed by the potent growth inhibitory effects of genetically or pharmacologically decreasing CaMKK2 activity; collectively, these findings suggest that CaMKK2 and CaMKIV may represent potential targets for hepatic cancer. PMID:25847065

  20. Measurement of true calcium absorption in premature infants using intravenous 46Ca and oral 44Ca.

    PubMed

    Hillman, L S; Tack, E; Covell, D G; Vieira, N E; Yergey, A L

    1988-06-01

    We have developed a method for measuring true fractional calcium absorption (alpha) in premature infants using two stable isotopes of calcium and tested it in seven studies in seven infants (birth weight 1543 +/- 65 g, gestation 32.8 +/- 7 wk). A total of 7.5 micrograms/kg 46Ca was given as a single intravenous bolus. Immediately thereafter 1.25 mg/kg of 44Ca was given in a single gavage feeding of standard infant formula (Enfamil). A metabolic isolette was used to obtain 4-h collections of urine for 24 h total. 46Ca and 44Ca were measured in urine by thermal ionization mass spectroscopy and expressed as the ratio to naturally occurring 48Ca. The differences in the 46Ca/48Ca and 44Ca/48Ca ratios from natural levels (delta % excess 46Ca and delta % excess 44Ca) were calculated. Percent absorption (alpha) equals a constant times cumulative delta % excess 44Ca/delta % excess 46Ca. The calculation of alpha is independent of urine volume or concentration. The delta % excess 46Ca, showed the expected multiexponential decline as a function of time, and delta % excess 44Ca usually peaked during a 4- to 8-h urine collection. Calculations of alpha using increasingly long sampling times showed that a plateau had been reached by 12 h. alpha values calculated after 16-24 h in the seven infants at 2 wk of age were 41, 48, 45, 46, 25, 55, and 51%. Repeat studies at 3 wk of age were 46, 60, and 54%. These values are somewhat higher than net percent calcium absorption values reported for standard formula and thus appear very appropriate.(ABSTRACT TRUNCATED AT 250 WORDS)

  1. A new method for 44Ca/40Ca determination using cool plasma MC-ICP-MS

    NASA Astrophysics Data System (ADS)

    Fietzke, J.; Eisenhauer, A.; Liebetrau, V.; Bock, B.; Gussone, N.; Nägler, T. F.; Dietzel, M.; Spero, H.; Bijma, J.; Dullo, C.

    2003-04-01

    Here we present a new technique for the direct measurement of 44Ca/40Ca isotope ratios on a MC-ICP-MS (Axiom) using the "cool plasma" technique. By reducing the plasma energy from about 1250 Watts to 400 Watts the isobaric effect resulting from 40Ar^+ can be significantly reduced enabling the simultaneous and precise measurement of 44Ca and 40Ca beam intensities in different Faraday cups. In contrast to the TIMS technique requiring a Ca double spike the isotope measurements on a MC-ICP-MS can be performed by bracketing standards. This reduces the effort for chemical preparations without loss of precision. Isobaric effects of MgO^+ and NaOH^+ interfering with 40Ca and MgOH_2 with 44Ca can be neglected by measuring Ca isotopes near the low mass edge of the peaks. No influences of Sr2+ were found monitoring on 43.5amu. Repeated measurements of two Johnson Matthey CaCO_3 standards (lot No. 4064 and lot No. 9912) revealed values of about -11.29 ppm and 0.57 ppm. These values are in accordance with previous values published by Russel et al. (1978) and Heuser et al. (2002). Repeated measurement of the NIST 915a CaCO_3 standard showed that the variance of a single δ44Ca measurement is about 0.28 ppm (2SD) being comparable with TIMS. MC-ICP-MS based δ44Ca values measured on inorganic precipitates are indistinguishable from earlier measurements of Gussone et al. (in press) based on TIMS δ44Ca measurements confirming that there is a positive δ44Ca-temperature gradient. Our study demonstrates the possibility to measure the whole dispersion of Ca isotopes with a MC-ICP-MS showing that 40Ca can be used for normalization of 44Ca. References Russel W. A. et al. (1978) Ca isotope fractionation on the Earth and other solar system materials. Geochim. Cosmochim. Acta 42, 1075--1090. Heuser A. et al. (2002) Measurement of Calcium Isotopes (δ44Ca) Using a Multicollector TIMS Technique. Int. J. Mass Spec. 220, 387--399. Gussone N. et al. (in press) Model for Kinetic Effects on

  2. Design and testing of Ground Penetrating Radar equipment dedicated for civil engineering applications: ongoing activities in Working Group 1 of COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Pajewski, Lara; Manacorda, Guido; Persico, Raffaele

    2015-04-01

    This work aims at presenting the ongoing research activities carried out in Working Group 1 'Novel GPR instrumentation' of the COST (European COoperation in Science and Technology) Action TU1208 'Civil Engineering Applications of Ground Penetrating Radar' (www.GPRadar.eu). The principal goal of the COST Action TU1208 is to exchange and increase scientific-technical knowledge and experience of GPR techniques in civil engineering, simultaneously promoting throughout Europe the effective use of this safe and non-destructive technique in the monitoring of infrastructures and structures. Working Group 1 (WG1) of the Action focuses on the development of innovative GPR equipment dedicated for civil engineering applications. It includes three Projects. Project 1.1 is focused on the 'Design, realisation and optimisation of innovative GPR equipment for the monitoring of critical transport infrastructures and buildings, and for the sensing of underground utilities and voids.' Project 1.2 is concerned with the 'Development and definition of advanced testing, calibration and stability procedures and protocols, for GPR equipment.' Project 1.3 deals with the 'Design, modelling and optimisation of GPR antennas.' During the first year of the Action, WG1 Members coordinated between themselves to address the state of the art and open problems in the scientific fields identified by the above-mentioned Projects [1, 2]. In carrying our this work, the WG1 strongly benefited from the participation of IDS Ingegneria dei Sistemi, one of the biggest GPR manufacturers, as well as from the contribution of external experts as David J. Daniels and Erica Utsi, sharing with the Action Members their wide experience on GPR technology and methodology (First General Meeting, July 2013). The synergy with WG2 and WG4 of the Action was useful for a deep understanding of the problems, merits and limits of available GPR equipment, as well as to discuss how to quantify the reliability of GPR results. An

  3. Ca/sup 2 +/-CaM-ATPase activity after corticosterone binding to synaptosomal plasma membrane

    SciTech Connect

    Iqbal, Z.; Sze, P.Y.

    1986-03-01

    Studies conducted in the laboratory have demonstrated that corticosterone (CS) binds specifically to synaptosomal plasma membrane (SPM) and modify the cellular events in the synaptosomes. On an exposure of a rat brain synaptosomes to physiological concentrations (<1..mu..M) of CS, the uptake of /sup 45/Ca/sup 2 +/ into synaptosomes was increased by 40-50%. Similarly the binding of /sup 45/Ca/sup 2 +/ to isolated SPM was also enhanced by 50% in the presence of CS. However when intact synaptosomes were pre-incubated with CS, the capacity of membranes to bind /sup 45/Ca/sup 2 +/ was increased by 2-fold. After the incubation of intact synaptosomes with < ..mu..M CS, the activity of trifluoperazine sensitive and CaM dependent Ca/sup 2 +/-ATPase at 1..mu..M Ca/sup 2 +/ was found to be stimulated by 20-30% whereas the activity of Ca/sup 2 +/-ATPase at 1mM Ca/sup 2 +/ remained unaffected. Exposure of synaptosomes to CS was also effective in protecting the reduction of ouabain-sensitive ATPase (Na/sup +/-K/sup +/-ATPase) activity caused by 0.5% ethanol used as a solvent medium for CS. These findings suggest that one of the physiological actions of CS in synaptosomes, after binding to synaptosomal plasma membranes, is an enhancement of Ca/sup 2 +/ transport and an increase of Ca/sup 2 +/-CaM-ATPase activity.

  4. Store-Operated Ca2+ Entry Sustains the Fertilization Ca2+ Signal in Pig Eggs.

    PubMed

    Wang, Chunmin; Zhang, Lu; Jaeger, Laurie A; Machaty, Zoltan

    2015-07-01

    The role of store-operated Ca(2+) entry (SOCE) in the maintenance of sperm-induced Ca(2+) oscillations was investigated in porcine eggs. We found that 10 μM gadolinium (Gd(3+)), which is known to inhibit SOCE, blocked Ca(2+) entry that was triggered by thapsigargin-induced store depletion and also caused an abrupt cessation of the fertilization Ca(2+) signal. In a similar manner 3,5-bis(trifluoromethyl)pyrazole 2 (20 μM), and tetrapandin-2 (10 μM), potent SOCE inhibitors, also blocked thapsigargin-stimulated Ca(2+) entry and disrupted the Ca(2+) oscillations after sperm-egg fusion. The downregulation of Stim1 or Orai1 in the eggs did not alter the Ca(2+) content of the intracellular stores, whereas co-overexpression of these proteins led to the generation of irregular Ca(2+) transients after fertilization that stopped prematurely. We also found that thapsigargin completely emptied the endoplasmic reticulum, and that the series of Ca(2+) transients stopped abruptly after the addition of thapsigargin to the fertilized eggs, indicating that the proper reloading of the intracellular stores is a prerequisite for the maintenance of the Ca(2+) oscillations. These data strengthen our previous findings that in porcine eggs SOCE is a major signaling cascade that is responsible for sustaining the repetitive Ca(2+) signal at fertilization.

  5. Ca2+ microdomains near plasma membrane Ca2+ channels: impact on cell function.

    PubMed

    Parekh, Anant B

    2008-07-01

    In eukaryotic cells, a rise in cytoplasmic Ca(2+) can activate a plethora of responses that operate on time scales ranging from milliseconds to days. Inherent to the use of a promiscuous signal like Ca(2+) is the problem of specificity: how can Ca(2+) activate some responses but not others? We now know that the spatial profile of the Ca(2+) signal is important Ca(2+) does not simply rise uniformly throughout the cytoplasm upon stimulation but can reach very high levels locally, creating spatial gradients. The most fundamental local Ca(2+) signal is the Ca(2+) microdomain that develops rapidly near open plasmalemmal Ca(2+) channels like voltage-gated L-type (Cav1.2) and store-operated CRAC channels. Recent work has revealed that Ca(2+) microdomains arising from these channels are remarkably versatile in triggering a range of responses that differ enormously in both temporal and spatial profile. Here, I delineate basic features of Ca(2+) microdomains and then describe how these highly local signals are used by Ca(2+)-permeable channels to drive cellular responses. PMID:18467365

  6. Extracellular Ca2+-sensing receptors--an overview.

    PubMed

    Chang, Wenhan; Shoback, Dolores

    2004-03-01

    Extracellular Ca2+-sensing receptors (CaRs) are the molecular basis by which specialized cells detect and respond to changes in the extracellular [Ca2+] ([Ca2+]o). CaRs belong to the family C of G-protein coupled receptors (GPCRs). Activation of CaRs triggers signaling pathways that modify numerous cell functions. Multiple ligands regulate the activation of CaRs including multivalent cations, L-amino acids, and changes in ionic strength and pH. CaRs in parathyroid cells play a central role in systemic Ca2+ homeostasis in terrestrial tetrapods. Mutations of the CaR gene in humans cause diseases in which serum and urine [Ca2+] and parathyroid hormone (PTH) levels are altered. CaR homologues are also expressed in organs critical to Ca2+ transport in ancient and modern fish, suggesting that similar receptors may have long been involved in Ca2+ homeostasis in lower vertebrates before parathyroid glands developed in terrestrial vertebrates. CaR mRNA and protein are also expressed in tissues not directly involved in Ca2+ homeostasis. This implies that there may be other biological roles for CaRs. Studies of CaR-knockout mice confirm the importance of CaRs in the parathyroid gland and kidney. The functions of CaRs in tissues other than kidney and parathyroid gland, however, remain to be elucidated.

  7. Optical properties of the giant dielectric material CaCu_3Ti_4O_12 and CdCu_3Ti_4O_12 \\unboldmath

    NASA Astrophysics Data System (ADS)

    Homes, Christopher

    2002-03-01

    The cubic perovskite-related oxide CaCu_3Ti_4O_12 has one of the largest dielectric constants at room temperature ever measured, ɛ_0~= 80 000, [ɛ_0≈ ɛ_1(ω arrow 0)]; ɛ0 drops by a factor of 1000 below about 100 K to ɛ_0~= 100, but curiously no change in the crystallographic structure is observed. The substitution of Cd for Ca results in a much lower value for the dielectric constant of ɛ_0~= 500, but the same general temperature dependence. The temperature-dependent reflectance of CaCu_3Ti_4O_12 and CdCu_3Ti_4O_12 has been measured over a wide frequency range, and the real and imaginary parts of the complex dielectric function were calculated from a Kramers-Kronig analysis. The optical properties are dominated by the unscreened lattice modes. There are a total of 11 infrared-active Tu modes expected; all 11 modes are observed in the CdCu_3Ti_4O_12 material, but only 10 are found in CaCu_3Ti_4O_12; the mode at ≈ 480 cm-1 is absent. The low frequency mode at ≈ 120 cm-1 in CaCu_3Ti_4O_12 shows an anomalous increase in oscillator strength at low temperature, in violation of the f-sum rule.(C.C. Homes et al.), Science 293, 673 (2001). The same behavior is found, although to a lesser extent, in the Cd material. A normal coordinate analysis of the vibrational modes indicates that the low-frequency mode involves primarily the Ca(Cd) atoms, as well as the Cu-O sublattice. The increase in strength of this mode is an indication that this bonding is becoming more ionic, implying that there is a redistribution of charge on this sublattice at low temperature. This suggests that the electric fields responsible for the large value of ɛ0 may originate not from the TiO6 octahedra, as is often seen in ferroelectrics, but from the Ca/Cu-O sublattice. A possible mechanism for the reduction of ɛ0 at low temperature will be discussed in light of recent high-resolution structural studies.

  8. Aging and CaMKII alter intracellular Ca2+ transients and heart rhythm in Drosophila melanogaster.

    PubMed

    Santalla, Manuela; Valverde, Carlos A; Harnichar, Ezequiel; Lacunza, Ezequiel; Aguilar-Fuentes, Javier; Mattiazzi, Alicia; Ferrero, Paola

    2014-01-01

    Aging is associated to disrupted contractility and rhythmicity, among other cardiovascular alterations. Drosophila melanogaster shows a pattern of aging similar to human beings and recapitulates the arrhythmogenic conditions found in the human heart. Moreover, the kinase CaMKII has been characterized as an important regulator of heart function and an arrhythmogenic molecule that participate in Ca2+ handling. Using a genetically engineered expressed Ca2+ indicator, we report changes in cardiac Ca2+ handling at two different ages. Aging prolonged relaxation, reduced spontaneous heart rate (HR) and increased the occurrence of arrhythmias, ectopic beats and asystoles. Alignment between Drosophila melanogaster and human CaMKII showed a high degree of conservation and indicates that relevant phosphorylation sites in humans are also present in the fruit fly. Inhibition of CaMKII by KN-93 (CaMKII-specific inhibitor), reduced HR without significant changes in other parameters. By contrast, overexpression of CaMKII increased HR and reduced arrhythmias. Moreover, it increased fluorescence amplitude, maximal rate of rise of fluorescence and reduced time to peak fluorescence. These results suggest that CaMKII in Drosophila melanogaster acts directly on heart function and that increasing CaMKII expression levels could be beneficial to improve contractility.

  9. Reduced endogenous Ca2+ buffering speeds active zone Ca2+ signaling.

    PubMed

    Delvendahl, Igor; Jablonski, Lukasz; Baade, Carolin; Matveev, Victor; Neher, Erwin; Hallermann, Stefan

    2015-06-01

    Fast synchronous neurotransmitter release at the presynaptic active zone is triggered by local Ca(2+) signals, which are confined in their spatiotemporal extent by endogenous Ca(2+) buffers. However, it remains elusive how rapid and reliable Ca(2+) signaling can be sustained during repetitive release. Here, we established quantitative two-photon Ca(2+) imaging in cerebellar mossy fiber boutons, which fire at exceptionally high rates. We show that endogenous fixed buffers have a surprisingly low Ca(2+)-binding ratio (∼ 15) and low affinity, whereas mobile buffers have high affinity. Experimentally constrained modeling revealed that the low endogenous buffering promotes fast clearance of Ca(2+) from the active zone during repetitive firing. Measuring Ca(2+) signals at different distances from active zones with ultra-high-resolution confirmed our model predictions. Our results lead to the concept that reduced Ca(2+) buffering enables fast active zone Ca(2+) signaling, suggesting that the strength of endogenous Ca(2+) buffering limits the rate of synchronous synaptic transmission. PMID:26015575

  10. Genetical and comparative genomics of Brassica under altered Ca supply identifies Arabidopsis Ca-transporter orthologs.

    PubMed

    Graham, Neil S; Hammond, John P; Lysenko, Artem; Mayes, Sean; O Lochlainn, Seosamh; Blasco, Bego; Bowen, Helen C; Rawlings, Chris J; Rios, Juan J; Welham, Susan; Carion, Pierre W C; Dupuy, Lionel X; King, Graham J; White, Philip J; Broadley, Martin R

    2014-07-01

    Although Ca transport in plants is highly complex, the overexpression of vacuolar Ca(2+) transporters in crops is a promising new technology to improve dietary Ca supplies through biofortification. Here, we sought to identify novel targets for increasing plant Ca accumulation using genetical and comparative genomics. Expression quantitative trait locus (eQTL) mapping to 1895 cis- and 8015 trans-loci were identified in shoots of an inbred mapping population of Brassica rapa (IMB211 × R500); 23 cis- and 948 trans-eQTLs responded specifically to altered Ca supply. eQTLs were screened for functional significance using a large database of shoot Ca concentration phenotypes of Arabidopsis thaliana. From 31 Arabidopsis gene identifiers tagged to robust shoot Ca concentration phenotypes, 21 mapped to 27 B. rapa eQTLs, including orthologs of the Ca(2+) transporters At-CAX1 and At-ACA8. Two of three independent missense mutants of BraA.cax1a, isolated previously by targeting induced local lesions in genomes, have allele-specific shoot Ca concentration phenotypes compared with their segregating wild types. BraA.CAX1a is a promising target for altering the Ca composition of Brassica, consistent with prior knowledge from Arabidopsis. We conclude that multiple-environment eQTL analysis of complex crop genomes combined with comparative genomics is a powerful technique for novel gene identification/prioritization.

  11. A Model-Independent Algorithm to Derive Ca2+ Fluxes Underlying Local Cytosolic Ca2+ Transients

    PubMed Central

    Ventura, Alejandra C.; Bruno, Luciana; Demuro, Angelo; Parker, Ian; Ponce Dawson, Silvina

    2005-01-01

    Local intracellular Ca2+ signals result from Ca2+ flux into the cytosol through individual channels or clusters of channels. To gain a mechanistic understanding of these events we need to know the magnitude and spatial distribution of the underlying Ca2+ flux. However, this is difficult to infer from fluorescence Ca2+ images because the distribution of Ca2+-bound dye is affected by poorly characterized processes including diffusion of Ca2+ ions, their binding to mobile and immobile buffers, and sequestration by Ca2+ pumps. Several methods have previously been proposed to derive Ca2+ flux from fluorescence images, but all require explicit knowledge or assumptions regarding these processes. We now present a novel algorithm that requires few assumptions and is largely model-independent. By testing the algorithm with both numerically generated image data and experimental images of sparklets resulting from Ca2+ flux through individual voltage-gated channels, we show that it satisfactorily reconstructs the magnitude and time course of the underlying Ca2+ currents. PMID:15681645

  12. Aging and CaMKII Alter Intracellular Ca2+ Transients and Heart Rhythm in Drosophila melanogaster

    PubMed Central

    Santalla, Manuela; Valverde, Carlos A.; Harnichar, Ezequiel; Lacunza, Ezequiel; Aguilar-Fuentes, Javier; Mattiazzi, Alicia; Ferrero, Paola

    2014-01-01

    Aging is associated to disrupted contractility and rhythmicity, among other cardiovascular alterations. Drosophila melanogaster shows a pattern of aging similar to human beings and recapitulates the arrhythmogenic conditions found in the human heart. Moreover, the kinase CaMKII has been characterized as an important regulator of heart function and an arrhythmogenic molecule that participate in Ca2+ handling. Using a genetically engineered expressed Ca2+ indicator, we report changes in cardiac Ca2+ handling at two different ages. Aging prolonged relaxation, reduced spontaneous heart rate (HR) and increased the occurrence of arrhythmias, ectopic beats and asystoles. Alignment between Drosophila melanogaster and human CaMKII showed a high degree of conservation and indicates that relevant phosphorylation sites in humans are also present in the fruit fly. Inhibition of CaMKII by KN-93 (CaMKII-specific inhibitor), reduced HR without significant changes in other parameters. By contrast, overexpression of CaMKII increased HR and reduced arrhythmias. Moreover, it increased fluorescence amplitude, maximal rate of rise of fluorescence and reduced time to peak fluorescence. These results suggest that CaMKII in Drosophila melanogaster acts directly on heart function and that increasing CaMKII expression levels could be beneficial to improve contractility. PMID:25003749

  13. Ca2+ signalling, voltage-gated Ca2+ channels and praziquantel in flatworm neuromusculature.

    PubMed

    Greenberg, R M

    2005-01-01

    Transient changes in calcium (Ca2+) levels regulate a wide variety of cellular processes, and cells employ both intracellular and extracellular sources of Ca2+ for signalling. Praziquantel, the drug of choice against schistosomiasis, disrupts Ca2+ homeostasis in adult worms. This review will focus on voltage-gated Ca2+ channels, which regulate levels of intracellular Ca2+ by coupling membrane depolarization to entry of extracellular Ca2+. Ca2+ channels are members of the ion channel superfamily and represent essential components of neurons, muscles and other excitable cells. Ca2+ channels are membrane protein complexes in which the pore-forming alpha1 subunit is modulated by auxiliary subunits such as beta and alpha2delta. Schistosomes express two Ca2+ channel beta subunit subtypes: a conventional subtype similar to beta subunits found in other vertebrates and invertebrates and a novel variant subtype with unusual structural and functional properties. The variant schistosome beta subunit confers praziquantel sensitivity to an otherwise praziquantel-insensitive mammalian Ca2+ channel, implicating it as a mediator of praziquantel action.

  14. Prognostic significance of preoperative serum CA125, CA19-9 and CEA in gastric carcinoma

    PubMed Central

    Wang, Wei; Chen, Xiao-Long; Zhao, Shen-Yu; Xu, Yu-Hui; Zhang, Wei-Han; Liu, Kai; Chen, Xin-Zu; Yang, Kun; Zhang, Bo; Chen, Zhi-Xin; Chen, Jia-Ping; Zhou, Zong-Guang; Hu, Jian-Kun

    2016-01-01

    The prognostic significance of preoperative serum CA125, CA19-9 and CEA in gastric carcinoma (GC) has been widely reported and is still under debate. Here, we evaluated the prognostic significance of preoperative serum CA125, CA19-9 and CEA in patients with GC. 1692 patients with GC who underwent gastrectomy were divided into the training (from January 2005 to December 2011, n = 1024) and the validation (from January 2012 to December 2013, n = 668) cohorts. Positive groups of CA125 (> 13.72 U/ml), CA19-9 (> 23.36 U/ml) and CEA (> 4.28 ng/ml) were significantly associated with more advanced clinicopathological traits and worse outcomes than that of negative groups (all P < 0.01). In Cox regression analysis, tumor size (P < 0.001, P = 0.005), pTNM stage (P < 0.001, P < 0.001) and CA125 (P = 0.026, P = 0.005) were independent prognostic factors both in two cohorts. Nomograms of these two cohorts based on the number of positive serum tumor markers (NPTM) were more accurate in prognostic prediction than TNM stage alone. Our findings suggested that elevated preoperative serum CA125, CA19-9 and CEA were associated with more advanced clinicopathological traits and less favorable outcomes. In addition, CA125 as an independent prognostic factor should be further investigated. Nomogram based on NPTM could accurately predict the prognosis of GC patients. PMID:27097114

  15. Effects of rapid buffers on Ca2+ diffusion and Ca2+ oscillations.

    PubMed Central

    Wagner, J; Keizer, J

    1994-01-01

    Based on realistic mechanisms of Ca2+ buffering that include both stationary and mobile buffers, we derive and investigate models of Ca2+ diffusion in the presence of rapid buffers. We obtain a single transport equation for Ca2+ that contains the effects caused by both stationary and mobile buffers. For stationary buffers alone, we obtain an expression for the effective diffusion constant of Ca2+ that depends on local Ca2+ concentrations. Mobile buffers, such as fura-2, BAPTA, or small endogenous proteins, give rise to a transport equation that is no longer strictly diffusive. Calculations are presented to show that these effects can modify greatly the manner and rate at which Ca2+ diffuses in cells, and we compare these results with recent measurements by Allbritton et al. (1992). As a prelude to work on Ca2+ waves, we use a simplified version of our model of the activation and inhibition of the IP3 receptor Ca2+ channel in the ER membrane to illustrate the way in which Ca2+ buffering can affect both the amplitude and existence of Ca2+ oscillations. PMID:7919018

  16. Regulated release of Ca2+ from respiring mitochondria by Ca2+/2H+ antiport.

    PubMed

    Fiskum, G; Lehninger, A L

    1979-07-25

    Simultaneous measurements of oxygen consumption and transmembrane transport of Ca2+, H+, and phosphate show that the efflux of Ca2+ from respiring tightly coupled rat liver mitochondria takes place by an electroneutral Ca2+/2H+ antiport process that is ruthenium red-insensitive and that is regulated by the oxidation-reduction state of the mitochondrial pyridine nucleotides. When mitochondrial pyridine nucleotides are kept in a reduced steady state, the efflux of Ca2+ is inhibited; when they are in an oxidized state, Ca2+ efflux is activated. These processes were demonstrated by allowing phosphate-depleted mitochondria respiring on succinate in the presence of rotenone to take up Ca2+ from the medium. Upon subsequent addition of ruthenium red to block Ca2+ transport via the electrophoretic influx pathway, and acetoacetate, to bring mitochondrial pyridine nucleotides into the oxidized state, Ca2+ efflux and H+ influx ensued. The observed H+ influx/Ca2+ efflux ratio was close to the value 2.0 predicted for the operation of an electrically neutral Ca2+/2H+ antiport process.

  17. Pericellular Ca2+ recycling potentiates thrombin-evoked Ca2+ signals in human platelets

    PubMed Central

    Sage, Stewart O; Pugh, Nicholas; Farndale, Richard W; Harper, Alan G S

    2013-01-01

    We have previously demonstrated that Na+/Ca2+ exchangers (NCXs) potentiate Ca2+ signaling evoked by thapsigargin in human platelets, via their ability to modulate the secretion of autocoids from dense granules. This link was confirmed in platelets stimulated with the physiological agonist, thrombin, and experiments were performed to examine how Ca2+ removal by the NCX modulates platelet dense granule secretion. In cells loaded with the near-membrane indicator FFP-18, thrombin stimulation was observed to elicit an NCX-dependent accumulation of Ca2+ in a pericellular region around the platelets. To test whether this pericellular Ca2+ accumulation might be responsible for the influence of NCXs over platelet function, platelets were exposed to fast Ca2+ chelators or had their glycocalyx removed. Both manipulations of the pericellular Ca2+ rise reduced thrombin-evoked Ca2+ signals and dense granule secretion. Blocking Ca2+-permeable ion channels had a similar effect, suggesting that Ca2+ exported into the pericellular region is able to recycle back into the platelet cytosol. Single cell imaging with extracellular Fluo-4 indicated that thrombin-evoked rises in extracellular [Ca2+] occurred within the boundary described by the cell surface, suggesting their presence within the open canalicular system (OCS). FFP-18 fluorescence was similarly distributed. These data suggest that upon thrombin stimulation, NCX activity creates a rise in [Ca2+] within the pericellular region of the platelet from where it recycles back into the platelet cytosol, acting to both accelerate dense granule secretion and maintain the initial rise in cytosolic [Ca2+]. PMID:24303163

  18. Calcium transport in bovine rumen epithelium as affected by luminal Ca concentrations and Ca sources

    PubMed Central

    Schröder, Bernd; Wilkens, Mirja R; Ricken, Gundula E; Leonhard-Marek, Sabine; Fraser, David R; Breves, Gerhard

    2015-01-01

    The quantitative role of different segments of the gastrointestinal tract for Ca absorption, the respective mechanisms, and their regulation are not fully identified for ruminants, that is, cattle. In different in vitro experiments the forestomach wall has been demonstrated to be a major site for active Ca absorption in sheep and goats. In order to further clarify the role of the bovine rumen for Ca transport with special attention to luminal Ca concentrations, its ionic form, and pH, electrophysiological and unidirectional flux rate measurements were performed with isolated bovine rumen epithelial tissues. For Ca flux studies (Jms, Jsm) in vitro Ussing chamber technique was applied. Standard RT-PCR method was used to characterize TRPV6 and PMCA1 as potential contributors to transepithelial active Ca transport. At Ca concentrations of 1.2 mmol L−1 on both sides of the tissues, Jms were higher than Jsm resulting under some conditions in significant Ca net flux rates (Jnet), indicating the presence of active Ca transport. In the absence of an electrical gradient, Jnet could significantly be stimulated in the presence of luminal short-chain fatty acids (SCFAs). Increasing the luminal Ca concentrations up to 11.2 mmol L−1 resulted in significant increases in Jms without influencing Jsm. Providing Ca in its form as respective chloride, formate, or propionate salts there was no significant effect on Jms. No transcripts specific for Ca channel TRPV6 could be demonstrated. Our results indicate different mechanisms for Ca absorption in bovine rumen as compared with those usually described for the small intestines. PMID:26564067

  19. Distribution of CA 125 in placental tissues.

    PubMed

    Fuith, L C; Müller-Holzner, E; Marth, C; Perkmann, E; Zeimet, A; Daxenbichler, G

    1989-01-01

    The presence of the tumor marker CA 125 was studied in different compartments of the human placenta. Levels of CA 125 in the cytosol of chorionic villi ranged from 27-17100 U/g (median 560 U/g). In the placental amnion and chorion concentrations ranged from 175-29000 U/g, median 1060 U/g and were not statistically different. In the umbilical cord values were significantly lower (range 44-7600 U/g; median 180 U/g). Maternal serum probes were above the upper limit of normal in all cases (range 48-500 U/ml; median 131 U/ml). Immunohistochemistry detected CA 125 exclusively within the amniotic cells of the placenta and the umbilical cord. This might be because CA 125 fixes more to insoluble structures in the amnion or because of contamination of chorionic villi with the underlying decidua.

  20. Departure gate of acidic Ca2+ confirmed

    PubMed Central

    Jentsch, Thomas J; Hoegg-Beiler, Maja B; Vogt, Janis

    2015-01-01

    More potent, but less known than IP3 that liberates Ca2+ from the ER, NAADP releases Ca2+ from acidic stores. The notion that TPC channels mediate this Ca2+ release was questioned recently by studies suggesting that TPCs are rather PI(3,5)P2-activated Na+ channels. Ruas et al (2015) now partially reconcile these views by showing that TPCs significantly conduct both cations and confirm their activation by both NAADP and PI(3,5)P2. They attribute the failure of others to observe TPC-dependent NAADP-induced Ca2+ release in vivo to inadequate mouse models that retain partial TPC function. PMID:26022292

  1. Role of Ca++ in Shoot Gravitropism. [avena

    NASA Technical Reports Server (NTRS)

    Rayle, D. L.

    1985-01-01

    A cornerstone in the argument that Ca(2+) levels may regulate growth is the finding the EGTA promotes straight growth. The usual explanation for these results is that Ca(2+) chelation from cell walls results in wall loosening and thus accelerated straight growth. The ability of frozen-thawed Avena coleoptile tissue (subjected to 15g tension) to extend in response to EGTA and Quin II was examined. The EGTA when applied in weakly buffered (i.e., 0.1mM) neutral solutions initiates rapid extension. When the buffer strength is increased, similar concentrations of EGTA produce no growth response. This implies when EGTA liberated protons are released upon Ca(2+) chelation they can either initiate acid growth (low buffer conditions) or if consumed (high buffer conditions) have no effect. Thus Ca(2+) chelation in itself apparently does not result in straight growth.

  2. Ca Isotope Fractionation in the Hawaiian Ecosystem

    NASA Astrophysics Data System (ADS)

    Wiegand, B. A.; Chadwick, O. A.; Vitousek, P. M.; Wooden, J. L.

    2003-12-01

    Investigations of the nutrient budgets in Hawaiian soils show the sources of major cations to be weathering of volcanic rock, marine aerosols, and Asian dust inputs. Especially at deeply weathered sites older than 150 ka, soils show strong depletion of the macronutrient calcium. Most of the calcium supply in these soils is of atmospheric origin (marine aerosols and continental dust). In contrast, younger soils are mainly supplied by calcium from weathering of volcanic bedrock. Based on the results of previous studies using strontium isotopic signatures and Sr/Ca ratios (e.g. Kennedy et al. 1998, Chadwick et al. 1999, Whipkey et al. 2000, Stewart et al. 2001) we have conducted research focusing on the isotope composition of calcium as a new tool for the investigation of sources of calcium and biogeochemical processes effecting Ca isotope fractionation in the plant-soil system. The study combines δ 44Ca with 87Sr/86Sr and Sr/Ca data of soils (bulk compositions and extractable Ca and Sr from soil exchange sites) and different plant species including native Ohia trees (Metrosideros polymorpha) from a soil chronosequence along the Hawaiian Island chain. The study sites differ in age of the underlying substrate from 0.3 ka to 4,100 ka, but show similar recent climate (mean annual temperature of 16 ° C) and amount of precipitation (about 2,500 mm/y). 44Ca/40Ca ratios were measured on a MAT262 at Stanford University, using a 42Ca-48Ca double spike, and are reported as δ 44Ca values relative to seawater (δ 44Ca = 0 ‰ ). Results of the extractable, plant available calcium from six soil sites show δ 44Ca values in the range of +1.2 ‰ to -1.3 ‰ with generally more negative values related to younger soil sites where calcium is mainly derived from weathering of volcanic rocks. Bulk soil samples, however, show δ 44Ca values between -0.1 ‰ and -2.5 ‰ , indicating differences in composition as a result of contributions from volcanic minerals, continental dust, and

  3. Ca channel gating during cardiac action potentials.

    PubMed

    Mazzanti, M; DeFelice, L J

    1990-10-01

    How do Ca channels conduct Ca ions during the cardiac action potential? We attempt to answer this question by applying a two-microelectrode technique, previously used for Na and K currents, in which we record the patch current and the action potential at the same time (Mazzanti, M., and L. J. DeFelice. 1987. Biophys. J. 12:95-100, and 1988. Biophys. J. 54:1139-1148; Wellis, D., L. J. DeFelice, and M. Mazzanti. 1990. Biophys. J. 57:41-48). In this paper, we also compare the action currents obtained by the technique with the step-protocol currents obtained during standard voltage-clamp experiments. Individual Ca channels were measured in 10 mM Ca/1 Ba and 10 mM Ba. To describe part of our results, we use the nomenclature introduced by Hess, P., J. B. Lansman, and R. W. Tsien (1984. Nature (Lond.). 311:538-544). With Ba as the charge carrier, Ca channel kinetics convert rapidly from long to short open times as the patch voltage changes from 20 to -20 mV. This voltage-dependent conversion occurs during action potentials and in step-protocol experiments. With Ca as the charge carrier, the currents are brief at all voltages, and it is difficult to define either the number of channels in the patch or the conductance of the individual channels. Occasionally, however, Ca-conducting channels spontaneously convert to long-open-time kinetics (in Hess et al., 1984, notation, mode 2). When this happens, which is about once in every 100beats, there usually appears to be only one channel in the patch. In this rare configuration, the channel is open long enough to measure its conductance in 10 Ca/ 1 Ba. The value is 8-10 pS, which is about half the conductance in Ba. Because the long openings occur so infrequently with Ca as the charge carrier, they contribute negligibly to the average Ca current at any particular time during an action potential. However, the total number of Ca ions entering during these long openings may be significant when compared to the number entering by the

  4. Ca channel gating during cardiac action potentials.

    PubMed

    Mazzanti, M; DeFelice, L J

    1990-10-01

    How do Ca channels conduct Ca ions during the cardiac action potential? We attempt to answer this question by applying a two-microelectrode technique, previously used for Na and K currents, in which we record the patch current and the action potential at the same time (Mazzanti, M., and L. J. DeFelice. 1987. Biophys. J. 12:95-100, and 1988. Biophys. J. 54:1139-1148; Wellis, D., L. J. DeFelice, and M. Mazzanti. 1990. Biophys. J. 57:41-48). In this paper, we also compare the action currents obtained by the technique with the step-protocol currents obtained during standard voltage-clamp experiments. Individual Ca channels were measured in 10 mM Ca/1 Ba and 10 mM Ba. To describe part of our results, we use the nomenclature introduced by Hess, P., J. B. Lansman, and R. W. Tsien (1984. Nature (Lond.). 311:538-544). With Ba as the charge carrier, Ca channel kinetics convert rapidly from long to short open times as the patch voltage changes from 20 to -20 mV. This voltage-dependent conversion occurs during action potentials and in step-protocol experiments. With Ca as the charge carrier, the currents are brief at all voltages, and it is difficult to define either the number of channels in the patch or the conductance of the individual channels. Occasionally, however, Ca-conducting channels spontaneously convert to long-open-time kinetics (in Hess et al., 1984, notation, mode 2). When this happens, which is about once in every 100beats, there usually appears to be only one channel in the patch. In this rare configuration, the channel is open long enough to measure its conductance in 10 Ca/ 1 Ba. The value is 8-10 pS, which is about half the conductance in Ba. Because the long openings occur so infrequently with Ca as the charge carrier, they contribute negligibly to the average Ca current at any particular time during an action potential. However, the total number of Ca ions entering during these long openings may be significant when compared to the number entering by the

  5. The CARRIBA project: Clouds, Aerosol, Radiation, and tuRbulence in the tRade wInd regime over BArbados

    NASA Astrophysics Data System (ADS)

    Wex, H.; Siebert, H.; Roberts, G.; Schmeissner, T.; Ditas, F.; Wehner, B.; Izaguirre, M. A.; Stratmann, F.

    2012-12-01

    The marine aerosol and its influence on cloud droplets in the trade wind cumuli were examined in two measurement campaigns on Barbados (CARRIBA: Clouds, Aerosol, Radiation, and tuRbulence in the tRade wInd regime over BArbados). The campaigns took place in November 2010 and April 2011 and lasted about a month, each. We measured in-situ data ground based and airborne, where for the latter ACTOS (Airborne Cloud Turbulence Observation System), a helicopter borne platform, was employed. Particle number size distributions (NSD), total particle number concentrations and concentrations of CCN (cloud condensation nuclei) measured on ground agreed well with those measured airborne on ACTOS in the marine boundary layer (up to about 500m). Therefore the measurement station at Ragged Point (Prospero et al., 2003) can be assumed to measure aerosol representative of the marine boundary layer. Throughout the two different measurement months, the aerosol could be divided into three distinctly different types, where the difference in total particle number concentrations (N) and CCN largely depended on the history of the aerosol within the last 10 days prior to arriving at Barbados, as could clearly be seen from back-trajectories. For times when N < 300 1/cm3, the Hoppel minimum at around 70nm could clearly be seen. N and CCN were comparably large for continentally influenced air masses (from Africa or north-eastern South America), where particles from biomass burning or desert dust could be found in the accumulation mode size range. A particularly large mode consisting of small particles of about 30-40nm was found for air masses arriving at Barbados from the north. Here, likely continental pollution from North America and/or precipitation over the ocean (i.e. a wash out of preexisting particles) favored new particle formation. Particle hygroscopicity was derived for the different air masses, and κ of 0.7 was determined on average. CCN concentrations derived from measured NSD

  6. A Nanoparticulate Ferritin-Core Mimetic Is Well Taken Up by HuTu 80 Duodenal Cells and Its Absorption in Mice Is Regulated by Body Iron12

    PubMed Central

    Latunde-Dada, Gladys O; Pereira, Dora IA; Tempest, Bethan; Ilyas, Hibah; Flynn, Angela C; Aslam, Mohamad F; Simpson, Robert J; Powell, Jonathan J

    2014-01-01

    Background: Iron (Fe) deficiency anemia remains the largest nutritional deficiency disorder worldwide. How the gut acquires iron from nano Fe(III), especially at the apical surface, is incompletely understood. Objective: We developed a novel Fe supplement consisting of nanoparticulate tartrate-modified Fe(III) poly oxo-hydroxide [here termed nano Fe(III)], which mimics the Fe oxide core of ferritin and effectively treats iron deficiency anemia in rats. Methods: We determined transfer to the systemic circulation of nano Fe(III) in iron-deficient and iron-sufficient outbread Swiss mouse strain (CD1) mice with use of 59Fe-labeled material. Iron deficiency was induced before starting the Fe-supplementation period through reduction of Fe concentrations in the rodent diet. A control group of iron-sufficient mice were fed a diet with adequate Fe concentrations throughout the study. Furthermore, we conducted a hemoglobin repletion study in which iron-deficient CD1 mice were fed for 7 d a diet supplemented with ferrous sulfate (FeSO4) or nano Fe(III). Finally, we further probed the mechanism of cellular acquisition of nano Fe(III) by assessing ferritin formation, as a measure of Fe uptake and utilization, in HuTu 80 duodenal cancer cells with targeted inhibition of divalent metal transporter 1 (DMT1) and duodenal cytochrome b (DCYTB) before exposure to the supplemented iron sources. Differences in gene expression were assessed by quantitative polymerase chain reaction. Results: Absorption (means ± SEMs) of nano Fe(III) was significantly increased in iron-deficient mice (58 ± 19%) compared to iron-sufficient mice (18 ± 17%) (P = 0.0001). Supplementation of the diet with nano Fe(III) or FeSO4 significantly increased hemoglobin concentrations in iron-deficient mice (170 ± 20 g/L, P = 0.01 and 180 ± 20 g/L, P = 0.002, respectively). Hepatic hepcidin mRNA expression reflected the nonheme-iron concentrations of the liver and was also comparable for both nano Fe(III)– and

  7. Efficient 41Ca measurements for biomedical applications

    NASA Astrophysics Data System (ADS)

    Vockenhuber, C.; Schulze-König, T.; Synal, H.-A.; Aeberli, I.; Zimmermann, M. B.

    2015-10-01

    We present the performance of 41Ca measurements using low-energy Accelerator Mass Spectrometry (AMS) at the 500 kV facility TANDY at ETH Zurich. We optimized the measurement procedure for biomedical applications where reliability and high sample throughput is required. The main challenge for AMS measurements of 41Ca is the interfering stable isobar 41K. We use a simplified sample preparation procedure to produce calcium fluoride (CaF2) and extract calcium tri-fluoride ions (CaF3-) ions to suppress the stable isobar 41K. Although 41K is not completely suppressed we reach 41Ca/40Ca background level in the 10-12 range which is adequate for biomedical studies. With helium as a stripper gas we can use charge state 2+ at high transmission (∼50%). The new measurement procedure with the approximately 10 × improved efficiency and the higher accuracy due to 41K correction allowed us to measure more than 600 samples for a large biomedical study within only a few weeks of measurement time.

  8. Intracellular Ca2+ signaling and preimplantation development.

    PubMed

    Armant, D Randall

    2015-01-01

    The key, versatile role of intracellular Ca2+ signaling during egg activation after fertilization has been appreciated for several decades. More recently, evidence has accumulated supporting the concept that cytoplasmic Ca2+ is also a major signaling nexus during subsequent development of the fertilized ovum. This chapter will review the molecular reactions that regulate intracellular Ca2+ levels and cell function, the role of Ca2+ signaling during egg activation and specific examples of repetitive Ca2+ signaling found throughout pre- and peri-implantation development. Many of the upstream and downstream pathways utilized during egg activation are also critical for specific processes that take place during embryonic development. Much remains to be done to elucidate the full complexity of Ca2+ signaling mechanisms in preimplantation embryos to the level of detail accomplished for egg activation. However, an emerging concept is that because this second messenger can be modulated downstream of numerous receptors and is able to bind and activate multiple cytoplasmic signaling proteins, it can help the coordination of development through up- and downstream pathways that change with each embryonic stage.

  9. Stellar neutron capture rates for /sup 46/Ca and /sup 48/Ca

    SciTech Connect

    Kaeppeller, F.; Walter, G.; Mathews, G.J.

    1985-04-01

    Stellar neutron capture rates for /sup 46/Ca and /sup 48/Ca have been measured by the activation technique. Both kT = 25 keV Maxwellian-like incident neutron spectra and non-Maxwellian higher energy spectra have been utilized to study the possible role of individual capture resonances. Maxwellian-averaged (kT = 30 keV) cross sections of 5.7 +- 0.5 and 0.95 +- 0.09 mb are derived for /sup 46/Ca and /sup 48/Ca, respectively. The possibility of a neutron capture origin for /sup 46/Ca and /sup 48/Ca is discussed in the light of these new cross sections, as well as a mechanism for the production of the observed isotopic anomalies in inclusion EK-1-4-1 from the Allende meteorite.

  10. Sarcolemmal Ca(2+)-entry through L-type Ca(2+) channels controls the profile of Ca(2+)-activated Cl(-) current in canine ventricular myocytes.

    PubMed

    Horváth, Balázs; Váczi, Krisztina; Hegyi, Bence; Gönczi, Mónika; Dienes, Beatrix; Kistamás, Kornél; Bányász, Tamás; Magyar, János; Baczkó, István; Varró, András; Seprényi, György; Csernoch, László; Nánási, Péter P; Szentandrássy, Norbert

    2016-08-01

    Ca(2+)-activated Cl(-) current (ICl(Ca)) mediated by TMEM16A and/or Bestrophin-3 may contribute to cardiac arrhythmias. The true profile of ICl(Ca) during an actual ventricular action potential (AP), however, is poorly understood. We aimed to study the profile of ICl(Ca) systematically under physiological conditions (normal Ca(2+) cycling and AP voltage-clamp) as well as in conditions designed to change [Ca(2+)]i. The expression of TMEM16A and/or Bestrophin-3 in canine and human left ventricular myocytes was examined. The possible spatial distribution of these proteins and their co-localization with Cav1.2 was also studied. The profile of ICl(Ca), identified as a 9-anthracene carboxylic acid-sensitive current under AP voltage-clamp conditions, contained an early fast outward and a late inward component, overlapping early and terminal repolarizations, respectively. Both components were moderately reduced by ryanodine, while fully abolished by BAPTA, but not EGTA. [Ca(2+)]i was monitored using Fura-2-AM. Setting [Ca(2+)]i to the systolic level measured in the bulk cytoplasm (1.1μM) decreased ICl(Ca), while application of Bay K8644, isoproterenol, and faster stimulation rates increased the amplitude of ICl(Ca). Ca(2+)-entry through L-type Ca(2+) channels was essential for activation of ICl(Ca). TMEM16A and Bestrophin-3 showed strong co-localization with one another and also with Cav1.2 channels, when assessed using immunolabeling and confocal microscopy in both canine myocytes and human ventricular myocardium. Activation of ICl(Ca) in canine ventricular cells requires Ca(2+)-entry through neighboring L-type Ca(2+) channels and is only augmented by SR Ca(2+)-release. Substantial activation of ICl(Ca) requires high Ca(2+) concentration in the dyadic clefts which can be effectively buffered by BAPTA, but not EGTA. PMID:27189885

  11. Interlaboratory study for coral Sr/Ca and other element/Ca ratio measurements

    NASA Astrophysics Data System (ADS)

    Hathorne, Ed C.; Gagnon, Alex; Felis, Thomas; Adkins, Jess; Asami, Ryuji; Boer, Wim; Caillon, Nicolas; Case, David; Cobb, Kim M.; Douville, Eric; deMenocal, Peter; Eisenhauer, Anton; Garbe-Schönberg, Dieter; Geibert, Walter; Goldstein, Steven; Hughen, Konrad; Inoue, Mayuri; Kawahata, Hodaka; Kölling, Martin; Cornec, Florence L.; Linsley, Braddock K.; McGregor, Helen V.; Montagna, Paolo; Nurhati, Intan S.; Quinn, Terrence M.; Raddatz, Jacek; Rebaubier, Hélène; Robinson, Laura; Sadekov, Aleksey; Sherrell, Rob; Sinclair, Dan; Tudhope, Alexander W.; Wei, Gangjian; Wong, Henri; Wu, Henry C.; You, Chen-Feng

    2013-09-01

    The Sr/Ca ratio of coral aragonite is used to reconstruct past sea surface temperature (SST). Twenty-one laboratories took part in an interlaboratory study of coral Sr/Ca measurements. Results show interlaboratory bias can be significant, and in the extreme case could result in a range in SST estimates of 7°C. However, most of the data fall within a narrower range and the Porites coral reference material JCp-1 is now characterized well enough to have a certified Sr/Ca value of 8.838 mmol/mol with an expanded uncertainty of 0.089 mmol/mol following International Association of Geoanalysts (IAG) guidelines. This uncertainty, at the 95% confidence level, equates to 1.5°C for SST estimates using Porites, so is approaching fitness for purpose. The comparable median within laboratory error is <0.5°C. This difference in uncertainties illustrates the interlaboratory bias component that should be reduced through the use of reference materials like the JCp-1. There are many potential sources contributing to biases in comparative methods but traces of Sr in Ca standards and uncertainties in reference solution composition can account for half of the combined uncertainty. Consensus values that fulfil the requirements to be certified values were also obtained for Mg/Ca in JCp-1 and for Sr/Ca and Mg/Ca ratios in the JCt-1 giant clam reference material. Reference values with variable fitness for purpose have also been obtained for Li/Ca, B/Ca, Ba/Ca, and U/Ca in both reference materials. In future, studies reporting coral element/Ca data should also report the average value obtained for a reference material such as the JCp-1.

  12. Mitochondrial calcium uniporter MCU supports cytoplasmic Ca2+ oscillations, store-operated Ca2+ entry and Ca2+-dependent gene expression in response to receptor stimulation.

    PubMed

    Samanta, Krishna; Douglas, Sophie; Parekh, Anant B

    2014-01-01

    Ca2+ flux into mitochondria is an important regulator of cytoplasmic Ca2+ signals, energy production and cell death pathways. Ca2+ uptake can occur through the recently discovered mitochondrial uniporter channel (MCU) but whether the MCU is involved in shaping Ca2+ signals and downstream responses to physiological levels of receptor stimulation is unknown. Here, we show that modest stimulation of leukotriene receptors with the pro-inflammatory signal LTC4 evokes a series of cytoplasmic Ca2+ oscillations that are rapidly and faithfully propagated into mitochondrial matrix. Knockdown of MCU or mitochondrial depolarisation, to reduce the driving force for Ca2+ entry into the matrix, prevents the mitochondrial Ca2+ rise and accelerates run down of the oscillations. The loss of cytoplasmic Ca2+ oscillations appeared to be a consequence of enhanced Ca2+-dependent inactivation of InsP3 receptors, which arose from the loss of mitochondrial Ca2+ buffering. Ca2+ dependent gene expression in response to leukotriene receptor activation was suppressed following knockdown of the MCU. In addition to buffering Ca2+ release, mitochondria also sequestrated Ca2+ entry through store-operated Ca2+ channels and this too was prevented following loss of MCU. MCU is therefore an important regulator of physiological pulses of cytoplasmic Ca2+.

  13. Regulation of RYR1 activity by Ca(2+) and calmodulin

    NASA Technical Reports Server (NTRS)

    Rodney, G. G.; Williams, B. Y.; Strasburg, G. M.; Beckingham, K.; Hamilton, S. L.

    2000-01-01

    The skeletal muscle calcium release channel (RYR1) is a Ca(2+)-binding protein that is regulated by another Ca(2+)-binding protein, calmodulin. The functional consequences of calmodulin's interaction with RYR1 are dependent on Ca(2+) concentration. At nanomolar Ca(2+) concentrations, calmodulin is an activator, but at micromolar Ca(2+) concentrations, calmodulin is an inhibitor of RYR1. This raises the question of whether the Ca(2+)-dependent effects of calmodulin on RYR1 function are due to Ca(2+) binding to calmodulin, RYR1, or both. To distinguish the effects of Ca(2+) binding to calmodulin from those of Ca(2+) binding to RYR1, a mutant calmodulin that cannot bind Ca(2+) was used to evaluate the effects of Ca(2+)-free calmodulin on Ca(2+)-bound RYR1. We demonstrate that Ca(2+)-free calmodulin enhances the affinity of RYR1 for Ca(2+) while Ca(2+) binding to calmodulin converts calmodulin from an activator to an inhibitor. Furthermore, Ca(2+) binding to RYR1 enhances its affinity for both Ca(2+)-free and Ca(2+)-bound calmodulin.

  14. The ryanodine receptor store-sensing gate controls Ca2+ waves and Ca2+-triggered arrhythmias.

    PubMed

    Chen, Wenqian; Wang, Ruiwu; Chen, Biyi; Zhong, Xiaowei; Kong, Huihui; Bai, Yunlong; Zhou, Qiang; Xie, Cuihong; Zhang, Jingqun; Guo, Ang; Tian, Xixi; Jones, Peter P; O'Mara, Megan L; Liu, Yingjie; Mi, Tao; Zhang, Lin; Bolstad, Jeff; Semeniuk, Lisa; Cheng, Hongqiang; Zhang, Jianlin; Chen, Ju; Tieleman, D Peter; Gillis, Anne M; Duff, Henry J; Fill, Michael; Song, Long-Sheng; Chen, S R Wayne

    2014-02-01

    Spontaneous Ca(2+) release from intracellular stores is important for various physiological and pathological processes. In cardiac muscle cells, spontaneous store overload-induced Ca(2+) release (SOICR) can result in Ca(2+) waves, a major cause of ventricular tachyarrhythmias (VTs) and sudden death. The molecular mechanism underlying SOICR has been a mystery for decades. Here we show that a point mutation, E4872A, in the helix bundle crossing region (the proposed gate) of the cardiac ryanodine receptor (RyR2) completely abolishes luminal, but not cytosolic, Ca(2+) activation of RyR2. The introduction of metal-binding histidines at this site converts RyR2 into a luminal Ni(2+)-gated channel. Mouse hearts harboring a heterozygous RyR2 mutation at this site (E4872Q) are resistant to SOICR and are completely protected against Ca(2+)-triggered VTs. These data show that the RyR2 gate directly senses luminal (store) Ca(2+), explaining the regulation of RyR2 by luminal Ca(2+), the initiation of Ca(2+) waves and Ca(2+)-triggered arrhythmias. This newly identified store-sensing gate structure is conserved in all RyR and inositol 1,4,5-trisphosphate receptor isoforms.

  15. Negative feedback from CaSR signaling to aquaporin-2 sensitizes vasopressin to extracellular Ca2.

    PubMed

    Ranieri, Marianna; Tamma, Grazia; Di Mise, Annarita; Russo, Annamaria; Centrone, Mariangela; Svelto, Maria; Calamita, Giuseppe; Valenti, Giovanna

    2015-07-01

    We previously described that high luminal Ca(2+) in the renal collecting duct attenuates short-term vasopressin-induced aquaporin-2 (AQP2) trafficking through activation of the Ca(2+)-sensing receptor (CaSR). Here, we evaluated AQP2 phosphorylation and permeability, in both renal HEK-293 cells and in the dissected inner medullary collecting duct, in response to specific activation of CaSR with NPS-R568. In CaSR-transfected cells, CaSR activation drastically reduced the basal levels of AQP2 phosphorylation at S256 (AQP2-pS256), thus having an opposite effect to vasopressin action. When forskolin stimulation was performed in the presence of NPS-R568, the increase in AQP2-pS256 and in the osmotic water permeability were prevented. In the freshly isolated inner mouse medullar collecting duct, stimulation with forskolin in the presence of NPS-R568 prevented the increase in AQP2-pS256 and osmotic water permeability. Our data demonstrate that the activation of CaSR in the collecting duct prevents the cAMP-dependent increase in AQP2-pS256 and water permeability, counteracting the short-term vasopressin response. By extension, our results suggest the attractive concept that CaSR expressed in distinct nephron segments exerts a negative feedback on hormones acting through cAMP, conferring high sensitivity of hormone to extracellular Ca(2+). PMID:25977473

  16. Radioisotope tracer studies of inorganic carbon and Ca in microbially derived CaCO3

    USGS Publications Warehouse

    Yates, Kimberly K.; Robbins, Lisa L.

    1999-01-01

    Microbial calcification significantly impacts the cycling and deposition of inorganic carbon. This research employs 45Ca and 14C techniques as radioisotopic tracers to examine the role of cellular cycling of Ca2+ and inorganic carbon in CaCO3 precipitation by the unicellular green alga Nannochloris atomus. Implications of the effects of these physiological aspects on CaCO3 precipitation and the effects of microbial calcification on CaCO3 δ13C ratios are discussed. Results from pulse/chase experiments indicate that intracellular Ca2+ is incorporated into extracellular CaCO3. Intracellular inorganic carbon leaks from cells within 10 to 12 s after injection of unlabelled NaHCO3, providing a source of inorganic carbon for extracellular CaCO3. Cellular expulsion of calcium plays a key role in increasing the CaCO3 saturation state at the site of calcification. The δ13C ratios of microbial carbonates may vary depending on the amount of photorespiratory CO2 incorporated.

  17. Pumping Ca2+ up H+ gradients: a Ca2+–H+ exchanger without a membrane

    PubMed Central

    Swietach, Pawel; Leem, Chae-Hun; Spitzer, Kenneth W; Vaughan-Jones, Richard D

    2014-01-01

    Cellular processes are exquisitely sensitive to H+ and Ca2+ ions because of powerful ionic interactions with proteins. By regulating the spatial and temporal distribution of intracellular [Ca2+] and [H+], cells such as cardiac myocytes can exercise control over their biological function. A well-established paradigm in cellular physiology is that ion concentrations are regulated by specialized, membrane-embedded transporter proteins. Many of these couple the movement of two or more ionic species per transport cycle, thereby linking ion concentrations among neighbouring compartments. Here, we compare and contrast canonical membrane transport with a novel type of Ca2+–H+ coupling within cytoplasm, which produces uphill Ca2+ transport energized by spatial H+ ion gradients, and can result in the cytoplasmic compartmentalization of Ca2+ without requiring a partitioning membrane. The mechanism, demonstrated in mammalian myocytes, relies on diffusible cytoplasmic buffers, such as carnosine, homocarnosine and ATP, to which Ca2+ and H+ ions bind in an apparently competitive manner. These buffer molecules can actively recruit Ca2+ to acidic microdomains, in exchange for the movement of H+ ions. The resulting Ca2+ microdomains thus have the potential to regulate function locally. Spatial cytoplasmic Ca2+–H+ exchange (cCHX) acts like a ‘pump’ without a membrane and may be operational in many cell types. PMID:24514908

  18. Mg/Ca of Continental Ostracode Shells

    NASA Astrophysics Data System (ADS)

    Ito, E.; Forester, R. M.; Marco-Barba, J.; Mezquita, F.

    2007-12-01

    Marine ionic chemistry is thought to remain constant. This, together with the belief that marine calcifiers partition Mg/Ca in a systematic manner as functions of temperature (and Mg/Ca) of water forms the basis of the Mg/Ca thermometer. In continental settings both of these assumptions are usually not true. Continental waters contain a wide variety of solutes in absolute and relative ion concentrations. Hence, waters with identical Mg/Ca may have very different concentrations of Mg and Ca and very different anions. Here we use two examples to focus on the effects of ion chemistry on Mg/Ca partitioning in continental ostracode shells and we ignore the complexities of solute evolution, which can change Mg/Ca over timescales of minutes to millennia. Palacios-Fest and Dettman (2001) conducted a monthly study of ,Cypridopsis vidua at El Yeso Lake in Sonora, Mexico. They established a relation between temperature and average shell Mg/Ca using regression analyses on averaged data. When their Mg/Ca-temperature relation is applied to monthly ,C. vidua data from Page Pond near Cleveland, Ohio, water temperatures of -8 to -1°C are obtained. The observed Mg/Ca ranges for El Yeso Lake (0.31 to 0.46) and Page Pond (0.33 to 0.46) are similar, as are their specific conductivities (700 to 850μS for El Yeso Lake; 400 to 600μS for Page Pond). However, [Ca] is 140-260 mg/L for El Yeso, but only 70-90 mg/L for Page Pond. Page Pond data, in fact, shows a good temperature shell Mg/Ca relation for .C. vidua, but the relation is different from that at El Yeso. Hence, shell Mg/Ca is a multi-valued, family of curves function of temperature and Mg/Ca of water that depends on the [Mg] and [Ca] values in water and perhaps other factors. Our second example comes from sites near Valencia, Spain and involves shell data for ,Cyprideis torosa, an estuarine ostracode that is tolerant of a wide range of salinity and can live in continental waters as long as the carbonate alkalinity to Ca ratio is

  19. Combined detection of CEA, CA 19-9, CA 242 and CA 50 in the diagnosis and prognosis of resectable gastric cancer.

    PubMed

    Tian, Shu-Bo; Yu, Jian-Chun; Kang, Wei-Ming; Ma, Zhi-Qiang; Ye, Xin; Cao, Zhan-Jiang; Yan, Chao

    2014-01-01

    Our aim was to investigate the value of combined detection of serum carcinoembryonic antigen (CEA), carbohydrate antigen (CA) 19-9, CA 242 and CA 50 in diagnosis and assessment of prognosis in consecutive gastric cancer patients. Clinical data including preoperative serum CEA, CA 19-9, CA 242, and CA 50 values and information on clinical pathological factors were collected and analyzed retrospectively. Univariate and multivariate survival analyses were used to explore the relationship between tumor markers and survival. Positive rates of tumor markers CEA, CA 19-9, CA 242 and CA 50 in the diagnosis of gastric cancer were 17.7, 17.1, 20.4 and 13.8%, respectively, and the positive rate for all four markers combined was 36.6%. Patients with elevated preoperative serum concentrations of CEA, CA 19-9, CA 242 and CA 50, had late clinical tumor stage and significantly poorer overall survival. Five-year survival rates in patients with elevated CEA, CA 19-9, CA 242 and CA 50 were 28.1, 25.8, 27.0 and 24.1%, respectively, compared with 55.0, 55.4, 56.4 and 54.5% in patients with these markers at normal levels (p<0.01). In multivariate Cox proportional hazards analyses, an elevated CA 242 level was determined to be an independent prognostic marker in gastric cancer patients. Combined detection of four tumor markers increased the positive rate for gastric cancer diagnosis. CA 242 showed higher diagnostic value and CA 50 showed lower diagnostic value. In resectable gastric carcinoma, preoperative CA 242 level was associated with disease stage, and was found to be a significant independent prognostic marker in gastric cancer patients.

  20. The destiny of Ca(2+) released by mitochondria.

    PubMed

    Takeuchi, Ayako; Kim, Bongju; Matsuoka, Satoshi

    2015-01-01

    Mitochondrial Ca(2+) is known to regulate diverse cellular functions, for example energy production and cell death, by modulating mitochondrial dehydrogenases, inducing production of reactive oxygen species, and opening mitochondrial permeability transition pores. In addition to the action of Ca(2+) within mitochondria, Ca(2+) released from mitochondria is also important in a variety of cellular functions. In the last 5 years, the molecules responsible for mitochondrial Ca(2+) dynamics have been identified: a mitochondrial Ca(2+) uniporter (MCU), a mitochondrial Na(+)-Ca(2+) exchanger (NCLX), and a candidate for a mitochondrial H(+)-Ca(2+) exchanger (Letm1). In this review, we focus on the mitochondrial Ca(2+) release system, and discuss its physiological and pathophysiological significance. Accumulating evidence suggests that the mitochondrial Ca(2+) release system is not only crucial in maintaining mitochondrial Ca(2+) homeostasis but also participates in the Ca(2+) crosstalk between mitochondria and the plasma membrane and between mitochondria and the endoplasmic/sarcoplasmic reticulum.

  1. Ultrastructural and immunohistochemical localization of plasma membrane Ca2+-ATPase 4 in Ca2+-transporting epithelia.

    PubMed

    Alexander, R Todd; Beggs, Megan R; Zamani, Reza; Marcussen, Niels; Frische, Sebastian; Dimke, Henrik

    2015-10-01

    Plasma membrane Ca(2+)-ATPases (PMCAs) participate in epithelial Ca(2+) transport and intracellular Ca(2+) signaling. The Pmca4 isoform is enriched in distal nephron isolates and decreased in mice lacking the epithelial transient receptor potential vanilloid 5 Ca(2+) channel. We therefore hypothesized that Pmca4 plays a significant role in transcellular Ca(2+) flux and investigated the localization and regulation of Pmca4 in Ca(2+)-transporting epithelia. Using antibodies directed specifically against Pmca4, we found it expressed only in the smooth muscle layer of mouse and human intestines, whereas pan-specific Pmca antibodies detected Pmca1 in lateral membranes of enterocytes. In the kidney, Pmca4 showed broad localization to the distal nephron. In the mouse, expression was most abundant in segments coexpressing the epithelial ransient receptor potential vanilloid 5 Ca(2+) channel. Significant, albeit lower, expression was also evident in the region encompassing the cortical thick ascending limbs, macula densa, and early distal tubules as well as smooth muscle layers surrounding renal vessels. In the human kidney, a similar pattern of distribution was observed, with the highest PMCA4 expression in Na(+)-Cl(-) cotransporter-positive tubules. Electron microscopy demonstrated Pmca4 localization in distal nephron cells at both the basolateral membrane and intracellular perinuclear compartments but not submembranous vesicles, suggesting rapid trafficking to the plasma membrane is unlikely to occur in vivo. Pmca4 expression was not altered by perturbations in Ca(2+) balance, pointing to a housekeeping function of the pump in Ca(2+)-transporting epithelia. In conclusion, Pmca4 shows a divergent expression pattern in Ca(2+)-transporting epithelia, inferring diverse roles for this isoform not limited to transepithelial Ca(2+) transport. PMID:26180241

  2. Characterization of conserved and novel microRNAs and their targets, including a TuMV-induced TIR-NBS-LRR class R gene-derived novel miRNA in Brassica.

    PubMed

    He, Xiang-Feng; Fang, Yuan-Yuan; Feng, Lei; Guo, Hui-Shan

    2008-07-01

    Nine conserved miRNA families and three potential novel miRNAs in Brassica rapa were identified from a small RNA library. The expression patterns of some conserved miRNAs had different tissue specificity in Brassica and Arabidopsis. One of the three potential miRNAs, named bra-miR1885, was verified as a true functional miRNA. It could be induced specifically by Turnip mosaic virus (TuMV) infection, and target TIR-NBS-LRR class disease-resistant transcripts for cleavage. Based on the hypothesis for de novo generation of new miRNA genes and the sequence similarity between bra-MIR1885 precursor loci and target transcript sequences, we suggest that bra-MIR1885 is a new miRNA gene that originated through inverted duplication events from TIR-NBS-LRR class disease-resistant protein-coding gene sequences, which became bra-miR1885 targets.

  3. Simultaneous production of H{sub 2} and O{sub 2} in closed vessels by marine cyanobacterium Anabaena sp. TU37-1 under high-cell-density conditions

    SciTech Connect

    Kumazawa, Shuzo; Asakawa, Hidenori

    1995-05-20

    A marine cyanobacterium, Anabaena sp. TU37-1, exhibited stable production of hydrogen and oxygen in closed vessels. About 8.4 and 4.3 mL (at atmospheric pressure) of hydrogen and oxygen accumulated, respectively, in flasks with 20 mL gas phase during 48 h incubation. Thus, concentration of H{sub 2} and O{sub 2} became 26 and 13% of the gas phase, respectively. Duration of hydrogen production was prolonged by the periodic gas replacement in the reaction vessel. The conversion efficiencies of photosynthetically active radiation (fluorescent light, 22 W/m{sup 2}) to hydrogen were 2.4 and 2.2% during the initial 12- and 24-h incubation periods respectively.

  4. Diffusion of Ca and Mg in Calcite

    SciTech Connect

    Cygan, R.T.; Fisler, D.K.

    1999-02-10

    The self-diffusion of Ca and the tracer diffusion of Mg in calcite have been experimentally measured using isotopic tracers of {sup 25}Mg and {sup 44}Ca. Natural single crystals of calcite were coated with a thermally-sputtered oxide thin film and then annealed in a CO{sub 2} gas at one atmosphere total pressure and temperatures from 550 to 800 C. Diffusion coefficient values were derived from the depth profiles obtained by ion microprobe analysis. The resultant activation energies for Mg tracer diffusion and Ca self-diffusion are respectively: E{sub a}(Mg) = 284 {+-} 74 kJ/mol and E{sub a}(Ca) = 271 {+-} 80 kJ/mol. For the temperature ranges in these experiments, the diffusion of Mg is faster than Ca. The results are generally consistent in magnitude with divalent cation diffusion rates obtained in previous studies and provide a means of interpreting the thermal histories of carbonate minerals, the mechanism of dolomitization, and other diffusion-controlled processes. The results indicate that cation diffusion in calcite is relatively slow and cations are the rate-limiting diffusing species for the deformation of calcite and carbonate rocks. Application of the calcite-dolomite geothermometer to metamorphic assemblages will be constrained by cation diffusion and cooling rates. The direct measurement of Mg tracer diffusion in calcite indicates that dolomitization is unlikely to be accomplished by Mg diffusion in the solid state but by a recrystallization process.

  5. CaF2:Yb laser ceramics

    NASA Astrophysics Data System (ADS)

    Akchurin, M. Sh.; Basiev, T. T.; Demidenko, A. A.; Doroshenko, M. E.; Fedorov, P. P.; Garibin, E. A.; Gusev, P. E.; Kuznetsov, S. V.; Krutov, M. A.; Mironov, I. A.; Osiko, V. V.; Popov, P. A.

    2013-01-01

    CaF2:Yb fluoride laser ceramics, prepared by hot-forming, exhibit the same optical properties as starting single crystals. Slope efficiency of the Сa0.95Yb0.05F2.05 is equal to 35% in the pulsed mode of laser operation. Decrease of ytterbium concentration in CaF2:Yb samples down to 3 mol.% resulted in the essential improvement of Сa0.97Yb0.03F2.03 thermal conductivity from 3.5 to 4.5 W/m K, but slightly decreased (down to 30%) slope efficiency of the samples under both pulsed and CW mode of operation. Alternative hot-pressing synthesis of CaF2:Yb fluoride laser ceramics provided materials with superior mechanical properties (microhardness Н = 3.2 GPa and fracture toughness К1С = 0.65 МPа m1/2) in comparison with hot-formed and/or single crystal CaF2:Yb specimens. For the first time, lasing has been observed for the novel aforementioned hot-pressed CaF2:Yb ceramics.

  6. Ca2+ signaling during vertebrate somitogenesis.

    PubMed

    Webb, Sarah E; Miller, Andrew L

    2006-07-01

    A variety of Ca2+ signals, in the form of intercellular pulses and waves, have been reported to be associated with the various sequential stages of somitogenesis: from convergent extension and the formation of the paraxial mesoderm; during the patterning of the paraxial mesoderm to establish segmental units; throughout the formation of the morphological boundaries that delineate the segmental units, and finally from within the maturing somites as they undergo subsequent development and differentiation. Due to both the technical challenges presented in imaging intact, developing embryos, and the subtle nature of the Ca2+ transients generated, they have proved to be difficult to visualize. However, a combination of cultured cell preparations and improvements in explant and whole embryo imaging techniques has begun to reveal a new and exciting class of developmental Ca2+ signals. In this chapter, we review the small, but expanding, number of reports in the literature and attempt to identify common characteristics of the somitogenic Ca2+ transients, such as their mode of generation, as well as their spatial and temporal features. This may help to elucidate the significance and function of these intriguing Ca2+ transients and thus integrate them into the complex signaling networks that orchestrate early developmental events. PMID:16787560

  7. Control of ciliary motility by Ca sup 2+ : Integration of Ca sup 2+ -dependent functions and targets for Ca sup 2+ action

    SciTech Connect

    Evans, T.C.

    1988-01-01

    To identify functions that regulate Ca{sup 2+}-induced ciliary reversal in Paramecium, mutants defective in terminating depolarization-induced backward swimming were selected. Six independent recessive mutations (k-shy) comprising two complementation groups, k-shyA and k-shyB, were identified. All mutants exhibited prolonged backward swimming in depolarizing solutions. Voltage clamp studies revealed that mutant Ca{sup 2+} current amplitudes were reduced, but could be restored to wild type levels by EGTA injection. The recovery of the mutant Ca{sup 2+} current from Ca{sup 2+}-dependent inactivation, and the decay of the Ca{sup 2+}-dependent K{sup +} and Ca{sup 2+}-dependent Na{sup +} currents after depolarization were slow in k-shy compared to wild type. To identify protein targets of Ca{sup 2+} action, ciliary proteins that interact with calmodulin (CaM) were characterized. With a {sup 125}I-CaM blot assay, several CaM-binding proteins were identified including axonemal, soluble, and membrane-bound polypeptides. Competitive displacement studies with unlabeled Paramecium CaM, bovine CaM, and troponinC suggested that both protein types bind CaM with high affinity and specificity. To examine the presence of CaM-binding sites in intact axonemes, a filtration binding assay was developed.

  8. Stability relations in the system CaSiO3-CaMnSi2O6-CaFeSi2O6

    NASA Astrophysics Data System (ADS)

    Abrecht, Jürgen

    1980-10-01

    In the system CaSiO3-CaMnSi2O6-CaFeSi2O6 extensive miscibility gaps between pyroxenoids and clinopyroxenes are observed. The miscibility gap between Mn-bustamite and Mn-wollastonite has been determined experimentally by a hydrothermal technique between 400° and 1200° C at P f= 2 kbar. Further experiments have been performed at P f=9 kbar, which revealed a shifting of the miscibility gap towards more Ca-rich compositions. The bustamite phase is stabilized by high pressures and the wollastonite structure is the stable phase at high temperatures. Similar phase relations as along the join CaSiO3-CaMnSi2O6 exist along the join CaSiO3-CaFeSi2O6 but with a more extensive two-phase field of bustamite-clinopyroxene. Possible phase relations along the joins CaSiO3-CaMnSi2O6, CaSiO3-CaFeSi2O6 and CaFeSi2O6-CaMnSi2O6 are given in temperature-composition diagrams for low pressures, based on natural and experimental data.

  9. Modeling the contributions of Ca2+ flows to spontaneous Ca2+ oscillations and cortical spreading depression-triggered Ca2+ waves in astrocyte networks.

    PubMed

    Li, Bing; Chen, Shangbin; Zeng, Shaoqun; Luo, Qingming; Li, Pengcheng

    2012-01-01

    Astrocytes participate in brain functions through Ca(2+) signals, including Ca(2+) waves and Ca(2+) oscillations. Currently the mechanisms of Ca(2+) signals in astrocytes are not fully clear. Here, we present a computational model to specify the relative contributions of different Ca(2+) flows between the extracellular space, the cytoplasm and the endoplasmic reticulum of astrocytes to the generation of spontaneous Ca(2+) oscillations (CASs) and cortical spreading depression (CSD)-triggered Ca(2+) waves (CSDCWs) in a one-dimensional astrocyte network. This model shows that CASs depend primarily on Ca(2+) released from internal stores of astrocytes, and CSDCWs depend mainly on voltage-gated Ca(2+) influx. It predicts that voltage-gated Ca(2+) influx is able to generate Ca(2+) waves during the process of CSD even after depleting internal Ca(2+) stores. Furthermore, the model investigates the interactions between CASs and CSDCWs and shows that the pass of CSDCWs suppresses CASs, whereas CASs do not prevent the generation of CSDCWs. This work quantitatively analyzes the generation of astrocytic Ca(2+) signals and indicates different mechanisms underlying CSDCWs and non-CSDCWs. Research on the different types of Ca(2+) signals might help to understand the ways by which astrocytes participate in information processing in brain functions.

  10. Method comparison for CA 15-3, CA 19-9, and CA 125 determination using the new LOCI technique of Dimension Vista 1500 and Immulite 2000 XPI.

    PubMed

    Zur, Berndt; Holdenrieder, Stefan; Albers, Eike; Walgenbach-Brünagel, Gisela; Stoffel-Wagner, Birgit

    2012-01-01

    We performed method comparison for the tumor markers CA 15-3, CA 19-9, and CA 125 measured by luminescent oxygen channeling immunoassay technology on the Dimension Vista 1500 and by classic luminescence technology on the Immulite 2000 XPI. Within-day and total imprecision were determined according to Clinical and Laboratory Standards Institute (CLSI) guidelines using three serum pools at different clinically relevant levels. In addition, parallel measurements on both systems were performed in a total of 738 routine serum samples (133 CA 15-3, 395 CA 19-9, and 210 CA 125). Total imprecision of serum pools for CA 15-3 ranged between 4.6% and 5.9%, for CA 19-9 between 4.4% and 7.8%, and for CA 125 between 3.3% and 4.3%. Marker values determined within the measurement range of both systems correlated well with each other (R = 0.88 for CA 15-3, R = 0.93 for CA 19-9, and R = 0.96 for CA 125). Slopes between the Vista and the Immulite method were 0.96 for CA 125, 0.72 for CA 15-3, and 0.87 for CA 19-9, indicating lower values for CA 15-3 and CA 19-9 when measured by the Vista method. This was particularly obvious for CA 19-9 levels in the lower measuring range of <100 U/mL (R = 0.85; slope 0.73).

  11. Sulfide capacity of CaO-CaF2-SiO2 slags

    NASA Astrophysics Data System (ADS)

    Susaki, Katsujiro; Maeda, Masafumi; Sano, Nobuo

    1990-02-01

    The sulfide capacity C S 2- = (pct S2-) · ( P O 2/ P S 2)1/2) of CaO-CaF2-SiO2 slags saturated with CaO, 3CaO · SiO2 or 2CaOSiO2 was determined at 1200 °C, 1250 °C, 1300 °C, and 1350 °C by equilibrating molten slag, molten silver, and CO-CO2 gas mixtures. Higher sulfide capacities were obtained for CaO-saturated slags. A drastic decrease was observed in those values when the ratio pct CaO/pct SiO2 is less than 2. The sulfur partition between carbon-saturated iron melts and presently investigated slags was calculated by using the sulfide capacities obtained and the activity coefficient of sulfur in carbon-saturated iron, which was also experimentally determined. For slags saturated with CaO, partitions of sulfur as high as 10,000 were obtained at 1300 °C and 1350 °C. Correlations between the sulfide capacity and other basicity indexes such as carbonate capacity and theoretical optical basicity were also discussed.

  12. Multifaceted plasma membrane Ca(2+) pumps: From structure to intracellular Ca(2+) handling and cancer.

    PubMed

    Padányi, Rita; Pászty, Katalin; Hegedűs, Luca; Varga, Karolina; Papp, Béla; Penniston, John T; Enyedi, Ágnes

    2016-06-01

    Plasma membrane Ca(2+) ATPases (PMCAs) are intimately involved in the control of intracellular Ca(2+) concentration. They reduce Ca(2+) in the cytosol not only by direct ejection, but also by controlling the formation of inositol-1,4,5-trisphosphate and decreasing Ca(2+) release from the endoplasmic reticulum Ca(2+) pool. In mammals four genes (PMCA1-4) are expressed, and alternative RNA splicing generates more than twenty variants. The variants differ in their regulatory characteristics. They localize into highly specialized membrane compartments and respond to the incoming Ca(2+) with distinct temporal resolution. The expression pattern of variants depends on cell type; a change in this pattern can result in perturbed Ca(2+) homeostasis and thus altered cell function. Indeed, PMCAs undergo remarkable changes in their expression pattern during tumorigenesis that might significantly contribute to the unbalanced Ca(2+) homeostasis of cancer cells. This article is part of a Special Issue entitled: Calcium and Cell Fate. Guest Editors: Jacques Haiech, Claus Heizmann, Joachim Krebs, Thierry Capiod and Olivier Mignen. PMID:26707182

  13. Intercellular Ca2+ Waves: Mechanisms and Function

    PubMed Central

    Sanderson, Michael J.

    2012-01-01

    Intercellular calcium (Ca2+) waves (ICWs) represent the propagation of increases in intracellular Ca2+ through a syncytium of cells and appear to be a fundamental mechanism for coordinating multicellular responses. ICWs occur in a wide diversity of cells and have been extensively studied in vitro. More recent studies focus on ICWs in vivo. ICWs are triggered by a variety of stimuli and involve the release of Ca2+ from internal stores. The propagation of ICWs predominately involves cell communication with internal messengers moving via gap junctions or extracellular messengers mediating paracrine signaling. ICWs appear to be important in both normal physiology as well as pathophysiological processes in a variety of organs and tissues including brain, liver, retina, cochlea, and vascular tissue. We review here the mechanisms of initiation and propagation of ICWs, the key intra- and extracellular messengers (inositol 1,4,5-trisphosphate and ATP) mediating ICWs, and the proposed physiological functions of ICWs. PMID:22811430

  14. Ca3Mn2O7.

    PubMed

    Guiblin, Nicolas; Grebille, Dominique; Leligny, Henri; Martin, Christine

    2002-01-01

    The tricalcium dimanganese heptaoxide (Ca3Mn2O7) member of the Ruddlesden-Popper series Ca(n+1)Mn(n)O(3n+1), i.e. with n = 2, was previously reported with an I-centred tetragonal lattice [a(t) = 3.68 and c(t) = 19.57 A] by Fawcett, Sunstrom, Greenblatt, Croft & Ramanujachary [Chem. Mater. (1998), 10, 3643-3651]. It is now found to be orthorhombic, with an A-centred lattice [a = 5.2347 (6), b = 5.2421 (2) and c = 19.4177 (19) A]. The structure has been refined in space group A2(1)am using X-ray single-crystal diffraction data and assuming the existence of twin domains related by the (1-10) plane. A comparison with the basic perovskite structure CaMnO3 (n = infinity) is proposed.

  15. Characterizing CA{sub 2} and CA{sub 6} using ELNES

    SciTech Connect

    Altay, A.; Carter, C.B.; Rulis, P.; Ching, W.-Y.; Arslan, I.; Guelguen, M.A.

    2010-08-15

    Calcium aluminates, compounds in the CaO-Al{sub 2}O{sub 3} phase system, are used in high-temperature cements and refractory oxides and have wide range of potential technological applications due to their interesting optical, electrical, thermal, and mechanical properties. They are used in both crystalline and glassy form; the glass is an isotropic material while the crystalline materials may be highly anisotropic. This paper will consider two particular crystalline materials, CA{sub 2} and CA{sub 6}, but the results should be applicable to all calcium aluminates. Although CA{sub 2} and CA{sub 6} crystals contain the same chemical species, Ca, Al, and O, the coordination and local environments of these species are different in the two structures and hence they show very different energy-loss near-edge structures (ELNES) when examined by electron energy-loss spectroscopy (EELS) in the TEM. The data obtained using ELNES can effectively provide a fingerprint for each compound and a map for their electronic structure. Once such fingerprints are obtained, they can be used to identify nano-sized particles/grains or material at interfaces and grain boundaries. In the present study, the local symmetry fingerprints for CA{sub 2} and CA{sub 6} structures are reported combining experimental spectra with electronic-structure calculations that allow the different features in the spectra to be interpreted. Al-L{sub 2,3} and O-K edge core-loss spectra from CA{sub 2} and CA{sub 6} were measured experimentally using electron energy-loss spectroscopy in a monochromated scanning transmission electron microscope. The near-edge structures were calculated for the different phases using the orthogonalized linear combination of atomic-orbitals method, and took account of core-hole interactions. It is shown that CA{sub 2} and CA{sub 6} structures exhibit distinctive experimental ELNES fingerprints so that these two phases can be separately identified even when present in small volumes

  16. IP3R, store-operated Ca2+ entry and neuronal Ca2+ homoeostasis in Drosophila.

    PubMed

    Chakraborty, Sumita; Hasan, Gaiti

    2012-02-01

    The IP3R (inositol 1,4,5-trisphosphate receptor) releases Ca2+ from the ER (endoplasmic reticulum) store upon binding to its ligand InsP3, which is thought to be generated by activation of certain membrane-bound G-protein-coupled receptors in Drosophila. Depletion of Ca2+ in the ER store also activates SOCE (store-operated Ca2+ entry) from the extracellular milieu across the plasma membrane, leading to a second rise in cytosolic Ca2+, which is then pumped back into the ER. The role of the IP3R and SOCE in mediating Ca2+ homoeostasis in neurons, their requirement in neuronal function and effect on neuronal physiology and as a consequence behaviour, are reviewed in the present article.

  17. Characterization of materials for Ca/CaCrO4 thermal batteries

    NASA Astrophysics Data System (ADS)

    Guidotti, R. A.; Reinhardt, F. W.; Poole, R. L.

    The performance of pelletized Ca/CaCrO4 thermal batteries is known to be sensitive to processing of the catholyte or DEB mix, which consists of CaCrO4 depolarized(D), KCl-LiCl eutectic electrolyte(E), and SiO2 binder(B). The chemical composition of the DEB mix affects the electrochemical behavior. Little work has been reported, however, for the characterization of DEB mixes in relation to their performance in Ca/CaCrO4 thermal batteries. Considerable variability of battery performance has also been observed when different lots of sheet calcium are used with the same DEB. The causes for this behavior remain elusive. In an effort to resolve these discrepancies in materials behavior, a study was carried out to characterize DEB powders and pellets and, to a lesser extent, sheet calcium with the primary objective of correlating observed battery performance to easily measured physical and chemical properties.

  18. Characterization of cathodic corrosion products in the Ca/CaCrO/sub 4/ thermal battery

    SciTech Connect

    Guidotti, R.A.; Reinhardt, F.W.; Venturini, E.L.; Rogers, J.W. Jr.; Cathey, W.N.

    1985-05-01

    Using thermal analysis techniques, we investigated the corrosion process resulting from the reaction of iron, nickel, and stainless steel (used as current collectors in Ca/CaCrO/sub 4/ thermal batteries) with CaCrO/sub 4/ dissolved in LiCl-KCl eutectic. The reaction product for iron was synthesized in bulk external to the battery and was characterized by chemical analysis, X-ray diffraction, Moessbauer spectroscopy, X-ray photoelectron spectroscopy, static magnetization, and electrical conductivity. This characterization provides a better understanding of the cathodic corrosion processes that occur in the Ca/CaCrO/sub 4/ thermal battery, and how the properties of the reaction layer at the catholyte-current collector interface influence battery performance.

  19. Characterization of cathodic corrosion products in the Ca/CaCrO4 thermal battery

    NASA Astrophysics Data System (ADS)

    Guidotti, R. A.; Reinhardt, F. W.; Venturini, E. L.; Rogers, J. W., Jr.; Cathey, W. N.

    1985-05-01

    Using thermal analysis techniques, the corrosion process resulting from the reaction of iron, nickel, and stainless steel (used as current collectors in Ca/CaCrO4 thermal batteries) with CaCrO4 dissolved in LiCl-KCl eutectic was investigated. The reaction product for iron was synthesized in bulk external to the battery and was characterized by chemical analysis, X-ray diffraction, Moessbauer spectroscopy, X-ray photoelectron spectroscopy, static magnetization, and electrical conductivity. This characterization provides a better understanding of the cathodic corrosion processes that occur in the Ca/CaCrO4 thermal battery, and how the properties of the reaction layer at the catholyte-current collector interface influence battery performance.

  20. Ca2+-induced Ca2+ Release in Chromaffin Cells Seen from inside the ER with Targeted Aequorin

    PubMed Central

    Alonso, Maria Teresa; Barrero, Maria José; Michelena, Pedro; Carnicero, Estela; Cuchillo, Inmaculada; García, Antonio G.; García-Sancho, Javier; Montero, Mayte; Alvarez, Javier

    1999-01-01

    The presence and physiological role of Ca2+-induced Ca2+ release (CICR) in nonmuscle excitable cells has been investigated only indirectly through measurements of cytosolic [Ca2+] ([Ca2+]c). Using targeted aequorin, we have directly monitored [Ca2+] changes inside the ER ([Ca2+]ER) in bovine adrenal chromaffin cells. Ca2+ entry induced by cell depolarization triggered a transient Ca2+ release from the ER that was highly dependent on [Ca2+]ER and sensitized by low concentrations of caffeine. Caffeine-induced Ca2+ release was quantal in nature due to modulation by [Ca2+]ER. Whereas caffeine released essentially all the Ca2+ from the ER, inositol 1,4,5-trisphosphate (InsP3)- producing agonists released only 60–80%. Both InsP3 and caffeine emptied completely the ER in digitonin-permeabilized cells whereas cyclic ADP-ribose had no effect. Ryanodine induced permanent emptying of the Ca2+ stores in a use-dependent manner after activation by caffeine. Fast confocal [Ca2+]c measurements showed that the wave of [Ca2+]c induced by 100-ms depolarizing pulses in voltage-clamped cells was delayed and reduced in intensity in ryanodine-treated cells. Our results indicate that the ER of chromaffin cells behaves mostly as a single homogeneous thapsigargin-sensitive Ca2+ pool that can release Ca2+ both via InsP3 receptors or CICR. PMID:9922451

  1. Kinetic studies of Ca2+ binding and Ca2+ clearance in the cytosol of adrenal chromaffin cells.

    PubMed Central

    Xu, T; Naraghi, M; Kang, H; Neher, E

    1997-01-01

    The Ca2+ binding kinetics of fura-2, DM-nitrophen, and the endogenous Ca2+ buffer, which determine the time course of Ca2+ changes after photolysis of DM-nitrophen, were studied in bovine chromaffin cells. The in vivo Ca2+ association rate constants of fura-2, DM-nitrophen, and the endogenous Ca2+ buffer were measured to be 5.17 x 10(8) M-1 s-1, 3.5 x 10(7) M-1 s-1, and 1.07 x 10(8) M-1 s-1, respectively. The endogenous Ca2+ buffer appeared to have a low affinity for Ca2+ with a dissociation constant around 100 microM. A fast Ca2+ uptake mechanism was also found to play a dominant role in the clearance of Ca2+ after flashes at high intracellular free Ca2+ concentrations ([Ca2+]), causing a fast [Ca2+]i decay within seconds. This Ca2+ clearance was identified as mitochondrial Ca2+ uptake. Its uptake kinetics were studied by analyzing the Ca2+ decay at high [Ca2+]i after flash photolysis of DM-nitrophen. The capacity of the mitochondrial uptake corresponds to a total cytosolic Ca2+ load of approximately 1 mM. Images FIGURE 2 FIGURE 5 FIGURE 6 FIGURE 7 FIGURE 8 PMID:9199815

  2. Anoxia-induced elevation of cytosolic Ca2+ concentration depends on different Ca2+ sources in rice and wheat protoplasts.

    PubMed

    Yemelyanov, Vladislav V; Shishova, Maria F; Chirkova, Tamara V; Lindberg, Sylvia M

    2011-08-01

    The anoxia-dependent elevation of cytosolic Ca(2+) concentration, [Ca(2+)](cyt), was investigated in plants differing in tolerance to hypoxia. The [Ca(2+)](cyt) was measured by fluorescence microscopy in single protoplasts loaded with the calcium-fluoroprobe Fura 2-AM. Imposition of anoxia led to a fast (within 3 min) significant elevation of [Ca(2+)](cyt) in rice leaf protoplasts. A tenfold drop in the external Ca(2+) concentration (to 0.1 mM) resulted in considerable decrease of the [Ca(2+)](cyt) shift. Rice root protoplasts reacted upon anoxia with higher amplitude. Addition of plasma membrane (verapamil, La(3+) and EGTA) and intracellular membrane Ca(2+)-channel antagonists (Li(+), ruthenium red and cyclosporine A) reduced the anoxic Ca(2+)-accumulation in rice. Wheat protoplasts responded to anoxia by smaller changes of [Ca(2+)](cyt). In wheat leaf protoplasts, the amplitude of the Ca(2+)-shift little depended on the external level of Ca(2+). Wheat root protoplasts were characterized by a small shift of [Ca(2+)](cyt) under anoxia. Plasmalemma Ca(2+)-channel blockers had little effect on the elevation of cytosolic Ca(2+) in wheat protoplasts. Intact rice seedlings absorbed Ca(2+) from the external medium under anoxic treatment. On the contrary, wheat seedlings were characterized by leakage of Ca(2+). Verapamil abolished the Ca(2+) influx in rice roots and Ca(2+) efflux from wheat roots. Anoxia-induced [Ca(2+)](cyt) elevation was high particularly in rice, a hypoxia-tolerant species. In conclusion, both external and internal Ca(2+) stores are important for anoxic [Ca(2+)](cyt) elevation in rice, whereas the hypoxia-intolerant wheat does not require external sources for [Ca(2+)](cyt) rise. Leaf and root protoplasts similarly responded to anoxia, independent of their organ origin.

  3. Materials compatibility during the chlorination of molten CaCl/sub 2/. CaO salts. [CaCl/sub 2/. CaO salt

    SciTech Connect

    Rense, C.E.C.; Fife, K.W.; Bowersox, D.F.; Ferran, M.D.

    1987-01-01

    As part of our effort to develop a semicontinuous PuO/sub 2/ reduction process, we are investigating promising materials for containing a 900/sup 0/C molten CaCl/sub 2/ . CaO chlorination reaction. We want the material to contain this reaction and to be reusable. We tested candidate materials in a simulated salt (no plutonium) using anhydrous HCl as the chlorinating agent. Data are presented on the performance of 36 metals and alloys, 9 ceramics, and 3 coatings.

  4. Novel antimigraineur dotarizine releases Ca2+ from caffeine-sensitive Ca2+ stores of chromaffin cells

    PubMed Central

    Novalbos, Jesús; Abad-Santos, Francisco; Zapater, Pedro; Alvarez, Javier; Alonso, María Teresa; Montero, Mayte; García, Antonio G

    1999-01-01

    The novel antimigraineur, dotarizine (30 μM), increased cytosolic Ca2+ concentration, [Ca2+]c, in fura-2-loaded bovine adrenal chromaffin cells. This increase was transient, reached a peak in about 2–5 min (0.53±0.07 μM; n=19) and then declined to basal levels over a further 5 min period.This transient rise of [Ca2+]c was mimicked by 1 μM thapsigargin and by 30 μM cyclopiazonic acid (CPA), but not by 30 μM flunarizine. Both thapsigargin and CPA occluded the effects of dotarizine and vice versa.All three compounds suppressed the transient [Ca2+]c rises induced by caffeine (10 mM, 10 s); blockade induced by thapsigargin was irreversible and that induced by CPA and dotarizine was reversible.Of the three compounds, only dotarizine blocked reversibly the [Ca2+]c spikes induced by short pulses of high K+ (70 mM, 5 s), suggesting that dotarizine blocks voltage-dependent Ca2+ channels but CPA and thapsigargin do not.Dotarizine caused a gradual and reversible depletion of endoplasmic reticulum (ER) Ca2+ in chromaffin cells transfected with ER-targeted aequorin. CPA had a similar effect.These data show that dotarizine shares with thapsigargin and CPA the ability to deplete Ca2+ in the ER; this novel action of dotarizine could be relevant to its prophylactic effects in migraine. Unlike thapsigargin and CPA, however, dotarizine additionally and reversibly blocks Ca2+ entry through voltage-dependent Ca2+ channels. PMID:10516641

  5. Growth rate effects on Mg/Ca and Sr/Ca ratios constrained by belemnite calcite

    NASA Astrophysics Data System (ADS)

    Vinzenz Ullmann, Clemens

    2016-04-01

    Multiple temperature proxies from single species are important to achieve robust palaeotemperature estimates. Besides the commonly employed oxygen isotope thermometer, also Mg/Ca and Sr/Ca ratios perform well as proxies for calcification temperature in the shells of some species. While salinity changes affect the ratios of earth alkaline elements much less than the δ18O thermometer, metabolic effects may exert a strong control on the expression of element ratios. Such effects are hard to study because biomineralization experiments have to overcome large intraspecific variability and can hardly ever isolate the controls of a single parameter on shell geochemistry. The unique geometry of the belemnite rostrum constitutes an exception to this rule. Its shape, large size, and the visibility of growth increments as bands enable the analysis of multiple, correlatable, high resolution geochemical profiles in a single fossil. The effects of the growth rate variability amongst these profiles on Mg/Ca and Sr/Ca ratios has been tested here. Within a specimen of Passaloteuthis bisulcata (Early Toarcian, Cleveland Basin, UK), Mg/Ca and Sr/Ca data were obtained from four profiles. With respect to growth rate in the first profile, which was taken as a reference, the relative growth rates in the remaining three profiles varied by a factor of 0.9 to 2.7. Results suggest that relative growth rate is linearly correlated with Mg/Ca and Sr/Ca, with a decrease of Mg/Ca by 8 % and increase of Sr/Ca by 6 % per 100 % increase in relative growth rate. The observed trends are consistent with abiogenic precipitation experiments and suggest that crystal precipitation rate exerts a significant, predictable control on the element distribution in biogenic calcite.

  6. Cardiac microvascular endothelial cells express a functional Ca+ -sensing receptor.

    PubMed

    Berra Romani, Roberto; Raqeeb, Abdul; Laforenza, Umberto; Scaffino, Manuela Federica; Moccia, Francesco; Avelino-Cruz, Josè Everardo; Oldani, Amanda; Coltrini, Daniela; Milesi, Veronica; Taglietti, Vanni; Tanzi, Franco

    2009-01-01

    The mechanism whereby extracellular Ca(2+) exerts the endothelium-dependent control of vascular tone is still unclear. In this study, we assessed whether cardiac microvascular endothelial cells (CMEC) express a functional extracellular Ca(2+)-sensing receptor (CaSR) using a variety of techniques. CaSR mRNA was detected using RT-PCR, and CaSR protein was identified by immunocytochemical analysis. In order to assess the functionality of the receptor, CMEC were loaded with the Ca(2+)-sensitive fluorochrome, Fura-2/AM. A number of CaSR agonists, such as spermine, Gd(3+), La(3+) and neomycin, elicited a heterogeneous intracellular Ca(2+) signal, which was abolished by disruption of inositol 1,4,5-trisphosphate (InsP(3)) signaling and by depletion of intracellular stores with cyclopiazonic acid. The inhibition of the Na(+)/Ca(2+) exchanger upon substitution of extracellular Na(+) unmasked the Ca(2+) signal triggered by an increase in extracellular Ca(2+) levels. Finally, aromatic amino acids, which function as allosteric activators of CaSR, potentiated the Ca(2+) response to the CaSR agonist La(3+). These data provide evidence that CMEC express CaSR, which is able to respond to physiological agonists by mobilizing Ca(2+) from intracellular InsP(3)-sensitive stores.

  7. Mitochondrial Ca(2+) uptake in skeletal muscle health and disease.

    PubMed

    Zhou, Jingsong; Dhakal, Kamal; Yi, Jianxun

    2016-08-01

    Muscle uses Ca(2+) as a messenger to control contraction and relies on ATP to maintain the intracellular Ca(2+) homeostasis. Mitochondria are the major sub-cellular organelle of ATP production. With a negative inner membrane potential, mitochondria take up Ca(2+) from their surroundings, a process called mitochondrial Ca(2+) uptake. Under physiological conditions, Ca(2+) uptake into mitochondria promotes ATP production. Excessive uptake causes mitochondrial Ca(2+) overload, which activates downstream adverse responses leading to cell dysfunction. Moreover, mitochondrial Ca(2+) uptake could shape spatio-temporal patterns of intracellular Ca(2+) signaling. Malfunction of mitochondrial Ca(2+) uptake is implicated in muscle degeneration. Unlike non-excitable cells, mitochondria in muscle cells experience dramatic changes of intracellular Ca(2+) levels. Besides the sudden elevation of Ca(2+) level induced by action potentials, Ca(2+) transients in muscle cells can be as short as a few milliseconds during a single twitch or as long as minutes during tetanic contraction, which raises the question whether mitochondrial Ca(2+) uptake is fast and big enough to shape intracellular Ca(2+) signaling during excitation-contraction coupling and creates technical challenges for quantification of the dynamic changes of Ca(2+) inside mitochondria. This review focuses on characterization of mitochondrial Ca(2+) uptake in skeletal muscle and its role in muscle physiology and diseases. PMID:27430885

  8. Gonadotropin-releasing hormone-induced Ca2+ transients in single identified gonadotropes require both intracellular Ca2+ mobilization and Ca2+ influx.

    PubMed Central

    Shangold, G A; Murphy, S N; Miller, R J

    1988-01-01

    We examined the effects of gonadotropin-releasing hormone (GnRH) on the intracellular free Ca2+ concentration ([Ca2+]i) in single rat anterior pituitary gonadotropes identified by a reverse hemolytic plaque assay. Concentrations of GnRH greater than 10 pM elicited increases in [Ca2+]i in identified cells but not in others. In contrast, depolarization induced by 50 mM K+ increased [Ca2+]i in all cells. Ca2+ transients induced by GnRH exhibited a complex time course. After an initial rapid rise, the [Ca2+]i fell to near basal levels only to be followed by a secondary extended rise and fall. Analysis of the Ca2+ transients on a rapid time base revealed that responses frequently consisted of several rapid oscillations in [Ca2+]i. Removal of extracellular Ca2+ or addition of the dihydropyridine Ca2+-channel blocker nitrendipine completely blocked the secondary rise in [Ca2+]i but had no effect whatsoever on the initial spike. Nitrendipine also blocked 50 mM K+-induced increases in [Ca2+]i in identified gonadotropes. The secondary rise induced by GnRH could be enhanced by a phorbol ester in a nitrendipine-sensitive fashion. Multiple spike responses to GnRH stimulation of the same cell could only be obtained if subsequent Ca2+ influx was permitted either by allowing a secondary rise to occur or by producing a Ca2+ transient by depolarizing the cells with 50 mM K+. It therefore appears that the response to GnRH consists of an initial phase of Ca2+ mobilization, probably mediated by inositol trisphosphate, and a subsequent phase of Ca2+ influx through nitrendipine-sensitive Ca2+ channels that may be activated by protein kinase C. The relative roles of these phases in the control of gonadotropin secretion are discussed. Images PMID:3045819

  9. Inhibitors of the Ca{sup 2+}/calmodulin-dependent protein kinase phosphatase family (CaMKP and CaMKP-N)

    SciTech Connect

    Sueyoshi, Noriyuki; Takao, Toshihiko; Nimura, Takaki; Sugiyama, Yasunori; Numano, Takamasa; Shigeri, Yasushi; Taniguchi, Takanobu; Kameshita, Isamu Ishida, Atsuhiko

    2007-11-23

    Ca{sup 2+}/calmodulin-dependent protein kinase phosphatase (CaMKP) and its nuclear isoform CaMKP-N are unique Ser/Thr protein phosphatases that negatively regulate the Ca{sup 2+}/calmodulin-dependent protein kinase (CaMK) cascade by dephosphorylating multifunctional CaMKI, II, and IV. However, the lack of specific inhibitors of these phosphatases has hampered studies on these enzymes in vivo. In an attempt to obtain specific inhibitors, we searched inhibitory compounds and found that Evans Blue and Chicago Sky Blue 6B served as effective inhibitors for CaMKP. These compounds also inhibited CaMKP-N, but inhibited neither protein phosphatase 2C, another member of PPM family phosphatase, nor calcineurin, a typical PPP family phosphatase. The minimum structure required for the inhibition was 1-amino-8-naphthol-4-sulfonic acid. When Neuro2a cells cotransfected with CaMKIV and CaMKP-N were treated with these compounds, the dephosphorylation of CaMKIV was strongly suppressed, suggesting that these compounds could be used as potent inhibitors of CaMKP and CaMKP-N in vivo as well as in vitro.

  10. Effects of glucagon and vasopressin on hepatic Ca2+ release.

    PubMed Central

    Kraus-Friedmann, N

    1986-01-01

    The effects of physiological levels of glucagon on Ca2+ efflux were examined in the perfused rat liver. Two methods were used to estimate Ca2+ efflux: prior labeling of the calcium pools with 45Ca2+ and measurement of perfusate Ca2+ with atomic absorption. According to both methods, glucagon administration at the physiological level evoked Ca2+ release. The released Ca2+ originated mostly from a carbonylcyanide p-trifluoromethoxyphenylhydrazone (FCCP)-depletable pool and also from an FCCP-insensitive pool from which Ca2+ could be released with A23187. Maximally effective doses of glucagon and vasopressin had no additive effect on Ca2+ release. Prior administration of vasopressin resulted in markedly reduced Ca2+ release by glucagon. These results indicate that glucagon releases Ca2+ from the same pool that vasopressin does. PMID:3466169

  11. L-type Ca2+ channels serve as a sensor of the SR Ca2+ for tuning the efficacy of Ca2+-induced Ca2+ release in rat ventricular myocytes

    PubMed Central

    Takamatsu, Hajime; Nagao, Taku; Ichijo, Hidenori; Adachi-Akahane, Satomi

    2003-01-01

    In cardiac excitation-contraction coupling, Ca2+-induced Ca2+ release (CICR) from ryanodine receptors (RyRs), triggered by Ca2+ entry through the nearby L-type Ca2+ channel, induces Ca2+-dependent inactivation (CDI) of the Ca2+ channel. Aiming at elucidating the physiological role of CDI produced by CICR (CICR-dependent CDI), we investigated the contribution of the CICR-dependent CDI to action potential (AP) waveform and the amount of Ca2+-influx through Ca2+ channels during AP in rat ventricular myocytes. The elimination of the CICR-dependent CDI, by depletion of the SR Ca2+ with thapsigargin, significantly prolonged AP duration (APD). APD changed in parallel with the magnitude of CICR during the recovery of the SR Ca2+ content after transient depletion by caffeine. Such CICR-dependent change of APD persisted under the highly Ca2+ buffered condition where the Ca2+ signalling was restricted to nanoscale domains. Blockers of the Ca2+-dependent Cl− channel or the BK channel did not affect AP waveform. The amount of Ca2+-influx through Ca2+ channels during the SR-depleted type AP waveform, measured in the SR-depleted myocyte, was increased by 40% over that during the SR-intact type AP waveform measured in the SR-intact myocyte. The protein kinase A stimulation further enhanced the Ca2+-influx during AP under the SR-depleted condition to 70% of that under the SR-intact condition. These results indicate that the CICR-dependent CDI of L-type Ca2+ channels, under control of the privileged cross-signalling between L-type Ca2+ channels and RyRs, play important roles for monitoring and tuning the SR Ca2+ content via changes of AP waveform and the amount of Ca2+-influx during AP in ventricular myocytes. PMID:14561825

  12. Detection of Ca(2+)-binding proteins by electrophoretic migration in the presence of Ca2+ combined with 45Ca2+ overlay of protein blots

    SciTech Connect

    Garrigos, M.; Deschamps, S.; Viel, A.; Lund, S.; Champeil, P.; Moller, J.V.; le Maire, M. , Gif-sur-Yvette )

    1991-04-01

    When high affinity Ca(2+)-binding proteins like calmodulin, or proteins with a high Ca(2+)-binding capacity like calsequestrin, underwent sodium dodecyl sulfate-gel electrophoresis in Laemmli systems, their electrophoretic migration rates were much higher in gels containing 1 mM Ca2+ than in gels containing ethylene glycol bis(beta-aminoethyl ether) N,N{prime}-tetraacetic acid (EGTA). Replacement of EGTA by Ca2+ in the gel, combined with the blotting of electrophoretically separated proteins on polyvinylidene difluoride membranes and subsequent 45Ca2+ overlay, proved a very effective means of detecting Ca(2+)-binding proteins. This combined approach is important since artifacts occur in both techniques when used separately. We found that the usual procedure of adding Ca2+ to the sample before electrophoresis without including it in the gel itself permitted the detection of only very high affinity Ca(2+)-binding proteins.

  13. 78 FR 60366 - California Disaster #CA-00212

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-01

    ... From the Federal Register Online via the Government Publishing Office SMALL BUSINESS ADMINISTRATION California Disaster CA-00212 AGENCY: U.S. Small Business Administration. ACTION: Notice. SUMMARY...: 06/24/2014. ADDRESSES: Submit completed loan applications to: U.S. Small Business...

  14. 77 FR 58901 - California Disaster #CA-00190

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-24

    ... From the Federal Register Online via the Government Publishing Office SMALL BUSINESS ADMINISTRATION California Disaster CA-00190 AGENCY: U.S. Small Business Administration. ACTION: Notice. SUMMARY... Application Deadline Date: 06/14/2013. ADDRESSES: Submit completed loan applications to: U.S. Small...

  15. SNL/CA Cultural Resources Management Plan.

    SciTech Connect

    Larsen, Barbara L.

    2005-11-01

    The SNL/CA Cultural Resources Management Plan satisfies the site's Environmental Management System requirement to promote long-term stewardship of cultural resources. The plan summarizes the cultural and historical setting of the site, identifies existing procedures and processes that support protection and preservation of resources, and outlines actions that would be initiated if cultural resources were discovered onsite in the future.3

  16. 78 FR 39821 - California Disaster #CA-00202

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-07-02

    ... ADMINISTRATION California Disaster CA-00202 AGENCY: U.S. Small Business Administration. ACTION: Notice. SUMMARY: This is a notice of an Administrative declaration of a disaster for the State of California dated 06/25... INFORMATION CONTACT: A. Escobar, Office of Disaster Assistance, U.S. Small Business Administration, 409...

  17. Hippocampal CA1 Ripples as Inhibitory Transients

    PubMed Central

    Krishnan, Giri P; Fellous, Jean-Marc; Bazhenov, Maxim

    2016-01-01

    Memories are stored and consolidated as a result of a dialogue between the hippocampus and cortex during sleep. Neurons active during behavior reactivate in both structures during sleep, in conjunction with characteristic brain oscillations that may form the neural substrate of memory consolidation. In the hippocampus, replay occurs within sharp wave-ripples: short bouts of high-frequency activity in area CA1 caused by excitatory activation from area CA3. In this work, we develop a computational model of ripple generation, motivated by in vivo rat data showing that ripples have a broad frequency distribution, exponential inter-arrival times and yet highly non-variable durations. Our study predicts that ripples are not persistent oscillations but result from a transient network behavior, induced by input from CA3, in which the high frequency synchronous firing of perisomatic interneurons does not depend on the time scale of synaptic inhibition. We found that noise-induced loss of synchrony among CA1 interneurons dynamically constrains individual ripple duration. Our study proposes a novel mechanism of hippocampal ripple generation consistent with a broad range of experimental data, and highlights the role of noise in regulating the duration of input-driven oscillatory spiking in an inhibitory network. PMID:27093059

  18. literacy.ca EXPRESS. October 2009

    ERIC Educational Resources Information Center

    Movement for Canadian Literacy, 2009

    2009-01-01

    This issue of "literacy.ca EXPRESS" focuses on the topic of promising practice. Promising or good practice and lessons learned are used to describe useful practices, approaches or ideas. Articles included in this issue: (1) Practitioner Profile: Meet Connie Jones; (2) Highlights from the LAN (Learner Advisory Network); (3) In the Works... Projects…

  19. literacy.ca EXPRESS. April 2010

    ERIC Educational Resources Information Center

    Movement for Canadian Literacy, 2010

    2010-01-01

    This issue of "literacy.ca EXPRESS" focuses on poverty. The articles included in this issue are: (1) Poverty Overview; (2) Tony's Story; (3) LAN (Learner Advisory Network) Member's Story (Dianne Smith); (4) Linking Adult Literacy to Poverty Reduction; (5) MCL (Movement for Canadian Literacy) Update; (6) Highlights from the LAN; (7) Good to Know...…

  20. literacy.ca EXPRESS. December 2010

    ERIC Educational Resources Information Center

    Canadian Literacy and Learning Network, 2010

    2010-01-01

    This issue of "literacy.ca EXPRESS" features new and exciting developments, updates and exciting new resources. Articles included in this issue are: (1) Introducing CLLN (Canadian Literacy and Learning Network)!; (2) Supporting Learner Leadership; (3) Involving Learners by Patricia Ashie; (4) Catching Confidence; (5) CALL (Committee of Adult…

  1. The study of skeletal calcium metabolism with 41Ca and 45Ca

    NASA Astrophysics Data System (ADS)

    Freeman, Stewart P. H. T.; Beck, Belinda; Bierman, June M.; Caffee, Marc W.; Heaney, Robert P.; Holloway, Leah; Marcus, Robert; Southon, John R.; Vogel, John S.

    2000-10-01

    The living skeleton can be labeled for life by the administration of radiologically trivial amounts of 41Ca tracer. After initial elimination of tracer from the readily exchangeable calcium pools subsequent skeletal calcium turnover maintains and modulates the urine 41Ca content. Uniquely, bone calcium metabolism may then be studied with tracer in near equilibrium with the body's calcium and resorbing calcium directly measured by accelerator mass spectrometry (AMS) of excreta. Our experiments with 25 41Ca labeled subjects demonstrate excellent diurnal stability and remarkable response to intervention of the urine signal. Thus the tracer method may prove a competitive means of measuring the effects of antiresorptive osteoporosis treatments, for therapy development or even clinical monitoring. Novel studies of long-term skeletal evolution are also possible. We realize that routinely administered short-lived calcium radiotracers contain 41Ca impurities and that thousands of experimental participants have been historically inadvertently 41Ca labeled. The 41Ca urine index might now rapidly further be characterized by contemporary measurements of these one-time subjects, and with their by now thoroughly skeleton-equilibrated tracer they might be ideal participants in other new experiments. We are also investigating 45Ca AMS. It may prove preferable to label the skeleton with this radiotracer already familiar to bioscientists, but new to AMS.

  2. Ca(2+)-activated chloride channel activity during Ca(2+) alternans in ventricular myocytes.

    PubMed

    Kanaporis, Giedrius; Blatter, Lothar A

    2016-11-01

    Cardiac alternans, defined beat-to-beat alternations in contraction, action potential (AP) morphology or cytosolic Ca transient (CaT) amplitude, is a high risk indicator for cardiac arrhythmias. We investigated mechanisms of cardiac alternans in single rabbit ventricular myocytes. CaTs were monitored simultaneously with membrane currents or APs recorded with the patch clamp technique. A strong correlation between beat-to-beat alternations of AP morphology and CaT alternans was observed. During CaT alternans application of voltage clamp protocols in form of pre-recorded APs revealed a prominent Ca(2+)-dependent membrane current consisting of a large outward component coinciding with AP phases 1 and 2, followed by an inward current during AP repolarization. Approximately 85% of the initial outward current was blocked by Cl(-) channel blocker DIDS or lowering external Cl(-) concentration identifying it as a Ca(2+)-activated Cl(-) current (ICaCC). The data suggest that ICaCC plays a critical role in shaping beat-to-beat alternations in AP morphology during alternans.

  3. TU-F-12A-06: BEST IN PHYSICS (IMAGING) - A Novel Catheter-Based Radionuclide Imaging System to Characterize Atherosclerotic Plaque

    SciTech Connect

    Zaman, R; Kosuge, H; Carpenter, C; Pratx, G; Sun, C; McConnell, M; Xing, L

    2014-06-15

    Purpose: Atherosclerosis underlies coronary artery diseases, the leading cause of death in the United States and worldwide. In this study, we developed a novel catheter-based radionuclide imaging (CRI) system to image 18F-fluorodeoxyglucose (18F-FDG), a radionuclide, a marker of vascular inflammation, in murine carotid arteries and characterized the system for spatial resolution from multiple scintillating materials. Methods: The catheter system includes 35 mm and 8 mm fixed focal length lenses, which are subsequently connected to a CMOS camera and fiber holder. The distal ferrule of an image bundle is terminated with a wide-angle lens. The novelty of this system is a scintillating balloon with a crystal tip in the front of the wide angle lens to image light from the decay of 18F-FDG emission signal. The scintillating balloon is fabricated from 1mL of silicone RTV catalyst mixed with 1 mL base and 50 mg/mL calcium fluoride doped with Europium (CaF2:Eu). To identify the optimal scintillating materials with respect to resolution, we calculated modulation transfer function (MTF) of Yttrium Aluminum Garnet doped with Cerium (YAG:Ce), anthracene, and CaF2:Eu phosphors using a thin line optical phantom (Fig. 1a-1b). Macrophage-rich FVB murine atherosclerotic carotid plaque model (n = 4) was used in ex vivo experiments. Confirmatory imaging was also performed by an external optical imaging system (IVIS-200). Results: Analysis of the different phosphors (Fig 1b) showed that CaF2:Eu enabled the best resolution of 1.2μm. The CRI system visualized 18F-FDG in atherosclerotic plaques (Fig. 1d). The ligated left carotid (LR) artery exhibited 4× higher 18F-FDG signal intensity compared to the non-ligated right carotid (negative control) artery (1.65×10{sup 2} ±4.07×10{sup 1} vs. 4.44×10{sup 1}±2.17×10{sup 0}, A.U., p = 0.005) and confirmed with IVIS-200 (Fig. 1d). Conclusion: This CRI system enables high-resolution and sensitive detection of 18F-FDG uptake by murine

  4. DA-6034 Induces [Ca(2+)]i Increase in Epithelial Cells.

    PubMed

    Yang, Yu-Mi; Park, Soonhong; Ji, Hyewon; Kim, Tae-Im; Kim, Eung Kweon; Kang, Kyung Koo; Shin, Dong Min

    2014-04-01

    DA-6034, a eupatilin derivative of flavonoid, has shown potent effects on the protection of gastric mucosa and induced the increases in fluid and glycoprotein secretion in human and rat corneal and conjunctival cells, suggesting that it might be considered as a drug for the treatment of dry eye. However, whether DA-6034 induces Ca(2+) signaling and its underlying mechanism in epithelial cells are not known. In the present study, we investigated the mechanism for actions of DA-6034 in Ca(2+) signaling pathways of the epithelial cells (conjunctival and corneal cells) from human donor eyes and mouse salivary gland epithelial cells. DA-6034 activated Ca(2+)-activated Cl(-) channels (CaCCs) and increased intracellular calcium concentrations ([Ca(2+)]i) in primary cultured human conjunctival cells. DA-6034 also increased [Ca(2+)]i in mouse salivary gland cells and human corneal epithelial cells. [Ca(2+)]i increase of DA-6034 was dependent on the Ca(2+) entry from extracellular and Ca(2+) release from internal Ca(2+) stores. Interestingly, these effects of DA-6034 were related to ryanodine receptors (RyRs) but not phospholipase C/inositol 1,4,5-triphosphate (IP3) pathway and lysosomal Ca(2+) stores. These results suggest that DA-6034 induces Ca(2+) signaling via extracellular Ca(2+) entry and RyRs-sensitive Ca(2+) release from internal Ca(2+) stores in epithelial cells.

  5. Calculated and measured [Ca(2+)] in buffers used to calibrate Ca(2+) macroelectrodes.

    PubMed

    McGuigan, John A S; Stumpff, Friederike

    2013-05-01

    The ionized concentration of calcium in physiological buffers ([Ca(2+)]) is normally calculated using either tabulated constants or software programs. To investigate the accuracy of such calculations, the [Ca(2+)] in EGTA [ethylene glycol-bis(β-aminoethylether)-N,N,N|,N|-tetraacetic acid], BAPTA [1,2-bis(o-aminophenoxy) ethane-N,N,N|,N|-tetraacetic acid], HEDTA [N-(2-hydroxyethyl)-ethylenediamine-N,N|,N|-triacetic acid], and NTA [N,N-bis(carboxymethyl)glycine] buffers was estimated using the ligand optimization method, and these measured values were compared with calculated values. All measurements overlapped in the pCa range of 3.51 (NTA) to 8.12 (EGTA). In all four buffer solutions, there was no correlation between measured and calculated values; the calculated values differed among themselves by factors varying from 1.3 (NTA) to 6.9 (EGTA). Independent measurements of EGTA purity and the apparent dissociation constants for HEDTA and NTA were not significantly different from the values estimated by the ligand optimization method, further substantiating the method. Using two calibration solutions of pCa 2.0 and 3.01 and seven buffers in the pCa range of 4.0-7.5, calibration of a Ca(2+) electrode over the pCa range of 2.0-7.5 became a routine procedure. It is proposed that such Ca(2+) calibration/buffer solutions be internationally defined and made commercially available to allow the precise measurement of [Ca(2+)] in biology.

  6. Voltage-gated Ca2+ influx and mitochondrial Ca2+ initiate secretion from Aplysia neuroendocrine cells.

    PubMed

    Hickey, C M; Groten, C J; Sham, L; Carter, C J; Magoski, N S

    2013-10-10

    Neuroendocrine secretion often requires prolonged voltage-gated Ca(2+) entry; however, the ability of Ca(2+) from intracellular stores, such as endoplasmic reticulum or mitochondria, to elicit secretion is less clear. We examined this using the bag cell neurons, which trigger ovulation in Aplysia by releasing egg-laying hormone (ELH) peptide. Secretion from cultured bag cell neurons was observed as an increase in plasma membrane capacitance following Ca(2+) influx evoked by a 5-Hz, 1-min train of depolarizing steps under voltage-clamp. The response was similar for step durations of ≥ 50 ms, but fell off sharply with shorter stimuli. The capacitance change was attenuated by replacing external Ca(2+) with Ba(2+), blocking Ca(2+) channels, buffering intracellular Ca(2+) with EGTA, disrupting synaptic protein recycling, or genetic knock-down of ELH. Regarding intracellular stores, liberating mitochondrial Ca(2+) with the protonophore, carbonyl cyanide-p-trifluoromethoxyphenyl-hydrazone (FCCP), brought about an EGTA-sensitive elevation of capacitance. Conversely, no change was observed to Ca(2+) released from the endoplasmic reticulum or acidic stores. Prior exposure to FCCP lessened the train-induced capacitance increase, suggesting overlap in the pool of releasable vesicles. Employing GTP-γ-S to interfere with endocytosis delayed recovery (presumed membrane retrieval) of the capacitance change following FCCP, but not the train. Finally, secretion was correlated with reproductive behavior, in that neurons isolated from animals engaged in egg-laying presented a greater train-induced capacitance elevation vs quiescent animals. The bag cell neuron capacitance increase is consistent with peptide secretion requiring high Ca(2+), either from influx or stores, and may reflect the all-or-none nature of reproduction.

  7. Voltage-gated Ca2+ influx and mitochondrial Ca2+ initiate secretion from Aplysia neuroendocrine cells.

    PubMed

    Hickey, C M; Groten, C J; Sham, L; Carter, C J; Magoski, N S

    2013-10-10

    Neuroendocrine secretion often requires prolonged voltage-gated Ca(2+) entry; however, the ability of Ca(2+) from intracellular stores, such as endoplasmic reticulum or mitochondria, to elicit secretion is less clear. We examined this using the bag cell neurons, which trigger ovulation in Aplysia by releasing egg-laying hormone (ELH) peptide. Secretion from cultured bag cell neurons was observed as an increase in plasma membrane capacitance following Ca(2+) influx evoked by a 5-Hz, 1-min train of depolarizing steps under voltage-clamp. The response was similar for step durations of ≥ 50 ms, but fell off sharply with shorter stimuli. The capacitance change was attenuated by replacing external Ca(2+) with Ba(2+), blocking Ca(2+) channels, buffering intracellular Ca(2+) with EGTA, disrupting synaptic protein recycling, or genetic knock-down of ELH. Regarding intracellular stores, liberating mitochondrial Ca(2+) with the protonophore, carbonyl cyanide-p-trifluoromethoxyphenyl-hydrazone (FCCP), brought about an EGTA-sensitive elevation of capacitance. Conversely, no change was observed to Ca(2+) released from the endoplasmic reticulum or acidic stores. Prior exposure to FCCP lessened the train-induced capacitance increase, suggesting overlap in the pool of releasable vesicles. Employing GTP-γ-S to interfere with endocytosis delayed recovery (presumed membrane retrieval) of the capacitance change following FCCP, but not the train. Finally, secretion was correlated with reproductive behavior, in that neurons isolated from animals engaged in egg-laying presented a greater train-induced capacitance elevation vs quiescent animals. The bag cell neuron capacitance increase is consistent with peptide secretion requiring high Ca(2+), either from influx or stores, and may reflect the all-or-none nature of reproduction. PMID:23876326

  8. Cenozoic seawater Sr/Ca evolution

    NASA Astrophysics Data System (ADS)

    Sosdian, Sindia M.; Lear, Caroline H.; Tao, Kai; Grossman, Ethan L.; O'Dea, Aaron; Rosenthal, Yair

    2012-10-01

    Records of seawater chemistry help constrain temporal variations in geochemical processes that impact the global carbon cycle and climate through Earth's history. Here we reconstruct Cenozoic seawater Sr/Ca (Sr/Casw) using fossil Conus and turritellid gastropod Sr/Ca. Combined with an oxygen isotope paleotemperature record from the same samples, the gastropod record suggests that Sr/Caswwas slightly higher in the Eocene (˜11.4 ± 3 mmol/mol) than today (˜8.54 mmol/mol) and remained relatively stable from the mid- to late Cenozoic. We compare our gastropod Cenozoic Sr/Casw record with a published turritellid gastropod Sr/Casw record and other published biogenic (benthic foraminifera, fossil fish teeth) and inorganic precipitate (calcite veins) Sr/Caswrecords. Once the uncertainties with our gastropod-derived Sr/Casw are taken into account the Sr/Casw record agrees reasonably well with biogenic Sr/Caswrecords. Assuming a seawater [Ca] history derived from marine evaporite inclusions, all biogenic-based Sr/Casw reconstructions imply decreasing seawater [Sr] through the Cenozoic, whereas the calcite vein Sr/Casw reconstruction implies increasing [Sr] through the Cenozoic. We apply a simple geochemical model to examine the implications of divergence among these seawater [Sr] reconstructions and suggest that the interpretation and uncertainties associated with the gastropod and calcite vein proxies need to be revisited. Used in conjunction with records of carbonate depositional fluxes, our favored seawater Sr/Ca scenarios point to a significant increase in the proportion of aragonite versus calcite deposition in shelf sediments from the Middle Miocene, coincident with the proliferation of coral reefs. We propose that this occurred at least 10 million years after the seawater Mg/Ca threshold was passed, and was instead aided by declining levels of atmospheric carbon dioxide.

  9. ASteCA: Automated Stellar Cluster Analysis

    NASA Astrophysics Data System (ADS)

    Perren, G. I.; Vázquez, R. A.; Piatti, A. E.

    2015-04-01

    We present the Automated Stellar Cluster Analysis package (ASteCA), a suit of tools designed to fully automate the standard tests applied on stellar clusters to determine their basic parameters. The set of functions included in the code make use of positional and photometric data to obtain precise and objective values for a given cluster's center coordinates, radius, luminosity function and integrated color magnitude, as well as characterizing through a statistical estimator its probability of being a true physical cluster rather than a random overdensity of field stars. ASteCA incorporates a Bayesian field star decontamination algorithm capable of assigning membership probabilities using photometric data alone. An isochrone fitting process based on the generation of synthetic clusters from theoretical isochrones and selection of the best fit through a genetic algorithm is also present, which allows ASteCA to provide accurate estimates for a cluster's metallicity, age, extinction and distance values along with its uncertainties. To validate the code we applied it on a large set of over 400 synthetic MASSCLEAN clusters with varying degrees of field star contamination as well as a smaller set of 20 observed Milky Way open clusters (Berkeley 7, Bochum 11, Czernik 26, Czernik 30, Haffner 11, Haffner 19, NGC 133, NGC 2236, NGC 2264, NGC 2324, NGC 2421, NGC 2627, NGC 6231, NGC 6383, NGC 6705, Ruprecht 1, Tombaugh 1, Trumpler 1, Trumpler 5 and Trumpler 14) studied in the literature. The results show that ASteCA is able to recover cluster parameters with an acceptable precision even for those clusters affected by substantial field star contamination. ASteCA is written in Python and is made available as an open source code which can be downloaded ready to be used from its official site.

  10. The Electronic Spectra of CaN2(+) and Ca(N2)2(+)

    NASA Technical Reports Server (NTRS)

    Rodriguez-Santiago, Luis; Bauschlicher, Charles W., Jr.; Arnold, James (Technical Monitor)

    1998-01-01

    The ground and low-lying electronic states of CaN2(+) are studied at several levels of theory. The results for the X(sup 2)Sigma(+) state and the excited (2)(sup 2)Pi state, arising from occupying the Ca 4p orbital, are in good agreement with experiment. The analogous states of Ca(N2)2(+) are studied using the same theoretical approaches, and predictions are made as to the changes caused by the addition of the second N2 ligand.

  11. A study of the low-lying states of CaAr + and CaKr +

    NASA Astrophysics Data System (ADS)

    Heinemann, Christoph; Koch, Wolfram; Partridge, Harry

    1998-04-01

    The spectroscopic constants of the ground 2Σ + states of CaAr + and CaKr + are determined using high quality ab initio methods. The computed binding energies are 789 and 1252 cm -1, respectively, in good agreement with the experimental determination of Pullins, Scurlock, Reddic and Duncan (J. Chem. Phys. 104 (1996) 7518). The much smaller CaKr + binding energy determined by Buthelezi, Bellert, Lewis and Brucat (Chem. Phys. Lett. 246 (1995) 145) is shown to be due to deficiencies in the method used to approximate the binding energy of the excited state.

  12. Ca2+-dependent facilitation of Cav1.3 Ca2+ channels by densin and Ca2+/calmodulin-dependent protein kinase II

    PubMed Central

    Jenkins, Meagan A.; Christel, Carl J.; Jiao, Yuxia; Abiria, Sunday; Kim, Kristin Y.; Usachev, Yuriy M.; Obermair, Gerald J.; Colbran, Roger J.; Lee, Amy

    2010-01-01

    Cav1 (L-type) channels and calmodulin-dependent protein kinase II (CaMKII) are key regulators of Ca2+ signaling in neurons. CaMKII directly potentiates the activity of Cav1.2 and Cav1.3 channels, but the underlying molecular mechanisms are incompletely understood. Here, we report that the CaMKII-associated protein, densin, is required for Ca2+-dependent facilitation of Cav1.3 channels. While neither CaMKII nor densin independently affect Cav1.3 properties in transfected HEK293T cells, the two together augment Cav1.3 Ca2+ currents during repetitive, but not sustained, depolarizing stimuli. Facilitation requires Ca2+, CaMKII activation and its association with densin, as well as densin binding to the Cav1.3 α1 subunit C-terminal domain. Cav1.3 channels and densin are targeted to dendritic spines in neurons and form a complex with CaMKII in the brain. Our results demonstrate a novel mechanism for Ca2+-dependent facilitation that may intensify postsynaptic Ca2+ signals during high-frequency stimulation. PMID:20392935

  13. Exercise training reverses myocardial dysfunction induced by CaMKIIδC overexpression by restoring Ca2+ homeostasis.

    PubMed

    Høydal, Morten A; Stølen, Tomas O; Kettlewell, Sarah; Maier, Lars S; Brown, Joan Heller; Sowa, Tomas; Catalucci, Daniele; Condorelli, Gianluigi; Kemi, Ole J; Smith, Godfrey L; Wisløff, Ulrik

    2016-07-01

    Several conditions of heart disease, including heart failure and diabetic cardiomyopathy, are associated with upregulation of cytosolic Ca(2+)/calmodulin-dependent protein kinase II (CaMKIIδC) activity. In the heart, CaMKIIδC isoform targets several proteins involved in intracellular Ca(2+) homeostasis. We hypothesized that high-intensity endurance training activates mechanisms that enable a rescue of dysfunctional cardiomyocyte Ca(2+) handling and thereby ameliorate cardiac dysfunction despite continuous and chronic elevated levels of CaMKIIδC CaMKIIδC transgenic (TG) and wild-type (WT) mice performed aerobic interval exercise training over 6 wk. Cardiac function was measured by echocardiography in vivo, and cardiomyocyte shortening and intracellular Ca(2+) handling were measured in vitro. TG mice had reduced global cardiac function, cardiomyocyte shortening (47% reduced compared with WT, P < 0.01), and impaired Ca(2+) homeostasis. Despite no change in the chronic elevated levels of CaMKIIδC, exercise improved global cardiac function, restored cardiomyocyte shortening, and reestablished Ca(2+) homeostasis to values not different from WT. The key features to explain restored Ca(2+) homeostasis after exercise training were increased L-type Ca(2+) current density and flux by 79 and 85%, respectively (P < 0.01), increased sarcoplasmic reticulum (SR) Ca(2+)-ATPase (SERCA2a) function by 50% (P < 0.01), and reduced diastolic SR Ca(2+) leak by 73% (P < 0.01), compared with sedentary TG mice. In conclusion, exercise training improves global cardiac function as well as cardiomyocyte function in the presence of a maintained high CaMKII activity. The main mechanisms of exercise-induced improvements in TG CaMKIIδC mice are mediated via increased L-type Ca(2+) channel currents and improved SR Ca(2+) handling by restoration of SERCA2a function in addition to reduced diastolic SR Ca(2+) leak. PMID:27231311

  14. Ca2+ signals regulate mitochondrial metabolism by stimulating CREB-mediated expression of the mitochondrial Ca2+ uniporter gene MCU.

    PubMed

    Shanmughapriya, Santhanam; Rajan, Sudarsan; Hoffman, Nicholas E; Zhang, Xueqian; Guo, Shuchi; Kolesar, Jill E; Hines, Kevin J; Ragheb, Jonathan; Jog, Neelakshi R; Caricchio, Roberto; Baba, Yoshihiro; Zhou, Yandong; Kaufman, Brett A; Cheung, Joseph Y; Kurosaki, Tomohiro; Gill, Donald L; Madesh, Muniswamy

    2015-03-03

    Cytosolic Ca2+ signals, generated through the coordinated translocation of Ca2+ across the plasma membrane (PM) and endoplasmic reticulum (ER) membrane, mediate diverse cellular responses. Mitochondrial Ca2+ is important for mitochondrial function, and when cytosolic Ca2+ concentration becomes too high, mitochondria function as cellular Ca2+ sinks. By measuring mitochondrial Ca2+ currents, we found that mitochondrial Ca2+ uptake was reduced in chicken DT40 B lymphocytes lacking either the ER-localized inositol trisphosphate receptor (IP3R), which releases Ca2+ from the ER, or Orai1 or STIM1, components of the PM-localized Ca2+ -permeable channel complex that mediates store-operated calcium entry (SOCE) in response to depletion of ER Ca2+ stores. The abundance of MCU, the pore-forming subunit of the mitochondrial Ca2+ uniporter, was reduced in cells deficient in IP3R, STIM1, or Orai1. Chromatin immunoprecipitation and promoter reporter analyses revealed that the Ca2+ -regulated transcription factor CREB (cyclic adenosine monophosphate response element-binding protein) directly bound the MCU promoter and stimulated expression. Lymphocytes deficient in IP3R, STIM1, or Orai1 exhibited altered mitochondrial metabolism, indicating that Ca2+ released from the ER and SOCE-mediated signals modulates mitochondrial function. Thus, our results showed that a transcriptional regulatory circuit involving Ca2+ -dependent activation of CREB controls the Ca2+ uptake capability of mitochondria and hence regulates mitochondrial metabolism.

  15. Phosphorylation and activation of nuclear Ca{sup 2+}/calmodulin-dependent protein kinase phosphatase (CaMKP-N/PPM1E) by Ca{sup 2+}/calmodulin-dependent protein kinase I (CaMKI)

    SciTech Connect

    Onouchi, Takashi; Sueyoshi, Noriyuki; Ishida, Atsuhiko; Kameshita, Isamu

    2012-06-15

    Highlights: Black-Right-Pointing-Pointer CaMKP-N/PPM1E underwent proteolytic processing and translocated to cytosol. Black-Right-Pointing-Pointer The proteolysis was effectively inhibited by the proteasome inhibitors. Black-Right-Pointing-Pointer Ser-480 of zebrafish CaMKP-N was phosphorylated by cytosolic CaMKI. Black-Right-Pointing-Pointer Phosphorylation-mimic mutants of CaMKP-N showed enhanced activity. Black-Right-Pointing-Pointer These results suggest that CaMKP-N is regulated by CaMKI. -- Abstract: Nuclear Ca{sup 2+}/calmodulin-dependent protein kinase phosphatase (CaMKP-N/PPM1E) is an enzyme that dephosphorylates and downregulates multifunctional Ca{sup 2+}/calmodulin-dependent protein kinases (CaMKs) as well as AMP-dependent protein kinase. In our previous study, we found that zebrafish CaMKP-N (zCaMKP-N) underwent proteolytic processing and translocated to cytosol in a proteasome inhibitor-sensitive manner. In the present study, we found that zCaMKP-N is regulated by phosphorylation at Ser-480. When zCaMKP-N was incubated with the activated CaMKI, time-dependent phosphorylation of the enzyme was observed. This phosphorylation was significantly reduced when Ser-480 was replaced by Ala, suggesting that CaMKI phosphorylates Ser-480 of zCaMKP-N. Phosphorylation-mimic mutants, S480D and S480E, showed higher phosphatase activities than those of wild type and S480A mutant in solution-based phosphatase assay using various substrates. Furthermore, autophosphorylation of CaMKII after ionomycin treatment was more severely attenuated in Neuro2a cells when CaMKII was cotransfected with the phosphorylation-mimic mutant of zCaMKP-N than with the wild-type or non-phosphorylatable zCaMKP-N. These results strongly suggest that phosphorylation of zCaMKP-N at Ser-480 by CaMKI activates CaMKP-N catalytic activity and thereby downregulates multifunctional CaMKs in the cytosol.

  16. Imaging intraorganellar Ca2+ at subcellular resolution using CEPIA

    PubMed Central

    Suzuki, Junji; Kanemaru, Kazunori; Ishii, Kuniaki; Ohkura, Masamichi; Okubo, Yohei; Iino, Masamitsu

    2014-01-01

    The endoplasmic reticulum (ER) and mitochondria accumulate Ca2+ within their lumens to regulate numerous cell functions. However, determining the dynamics of intraorganellar Ca2+ has proven to be difficult. Here we describe a family of genetically encoded Ca2+ indicators, named calcium-measuring organelle-entrapped protein indicators (CEPIA), which can be utilized for intraorganellar Ca2+ imaging. CEPIA, which emit green, red or blue/green fluorescence, are engineered to bind Ca2+ at intraorganellar Ca2+ concentrations. They can be targeted to different organelles and may be used alongside other fluorescent molecular markers, expanding the range of cell functions that can be simultaneously analysed. The spatiotemporal resolution of CEPIA makes it possible to resolve Ca2+ import into individual mitochondria while simultaneously measuring ER and cytosolic Ca2+. We have used these imaging capabilities to reveal differential Ca2+ handling in individual mitochondria. CEPIA imaging is a useful new tool to further the understanding of organellar functions. PMID:24923787

  17. Ca2+ dialogue between acidic vesicles and ER.

    PubMed

    Morgan, Anthony J

    2016-04-15

    Extracellular stimuli evoke the synthesis of intracellular second messengers, several of which couple to the release of Ca(2+)from Ca(2+)-storing organelles via activation of cognate organellar Ca(2+)-channel complexes. The archetype is the inositol 1,4,5-trisphosphate (IP3) and IP3receptor (IP3R) on the endoplasmic reticulum (ER). A less understood, parallel Ca(2+)signalling cascade is that involving the messenger nicotinic acid adenine dinucleotide phosphate (NAADP) that couples to Ca(2+)release from acidic Ca(2+)stores [e.g. endo-lysosomes, secretory vesicles, lysosome-related organelles (LROs)]. NAADP-induced Ca(2+)release absolutely requires organellar TPCs (two-pore channels). This review discusses how ER and acidic Ca(2+)stores physically and functionally interact to generate and shape global and local Ca(2+)signals, with particular emphasis on the two-way dialogue between these two organelles.

  18. Coupling Ratio for Ca(2+) Transport by Calcium Oxalate Precipitation.

    PubMed

    Sehgal, Pankaj; Olesen, Claus; Møller, Jesper V

    2016-01-01

    The SERCA isoform 1a is constructed to transport 2 Ca(2+) ions across the sarcoplasmic reticulum membrane coupled to the hydrolysis of one molecule of MgATP. However, observed coupling ratios for Ca(2+) transported/ATP hydrolzyed are usually less than 2:1, since part of the Ca(2+) accumulated at high intravesicular concentrations by the active transport of Ca(2+) leaks out of the vesicles because of Ca(2+)-induced Ca(2+) exchange. However, in the presence of a high concentration of oxalate (5 mM) Ca(2+) will precipitate as Ca-oxalate inside the vesicles and thereby be prevented from leaking out and, in addition, this treatment will reduce the intravesicular free concentration of Ca(2+) to a level where optimal coupling ratios of 2:1 can be achieved.

  19. Generation and Behavior Characterization of CaMKIIβ Knockout Mice

    PubMed Central

    Tu, Tao; Goulding, Danielle S.; Haiech, Jacques; Watterson, D. Martin; Van Eldik, Linda J.

    2014-01-01

    The calcium/calmodulin-dependent protein kinase II (CaMKII) is abundant in the brain, where it makes important contributions to synaptic organization and homeostasis, including playing an essential role in synaptic plasticity and memory. Four genes encode isoforms of CaMKII (α, β, δ, γ), with CaMKIIα and CaMKIIβ highly expressed in the brain. Decades of molecular and cellular research, as well as the use of a large number of CaMKIIα mutant mouse lines, have provided insight into the pivotal roles of CaMKIIα in brain plasticity and cognition. However, less is known about the CaMKIIβ isoform. We report the development and extensive behavioral and phenotypic characterization of a CaMKIIβ knockout (KO) mouse. The CaMKIIβ KO mouse was found to be smaller at weaning, with an altered body mass composition. The CaMKIIβ KO mouse showed ataxia, impaired forelimb grip strength, and deficits in the rotorod, balance beam and running wheel tasks. Interestingly, the CaMKIIβ KO mouse exhibited reduced anxiety in the elevated plus maze and open field tests. The CaMKIIβ KO mouse also showed cognitive impairment in the novel object recognition task. Our results provide a comprehensive behavioral characterization of mice deficient in the β isoform of CaMKII. The neurologic phenotypes and the construction of the genotype suggest the utility of this KO mouse strain for future studies of CaMKIIβ in brain structure, function and development. PMID:25127391

  20. Fabrication aspects of PLA-CaP/PLGA-CaP composites for orthopedic applications: a review.

    PubMed

    Zhou, Huan; Lawrence, Joseph G; Bhaduri, Sarit B

    2012-07-01

    For several decades, composites made of polylactic acid-calcium phosphates (PLA-CaP) and polylactic acid-co-glycolic acid-calcium phosphates (PLGA-CaP) have seen widespread uses in orthopedic applications. This paper reviews the fabrication aspects of these composites, following the ubiquitous materials science approach by studying "processing-structure-property" correlations. Various fabrication processes such as microencapsulation, phase separation, electrospinning, supercritical gas foaming, etc., are reviewed, with specific examples of their applications in fabricating these composites. The effect of the incorporation of CaP materials on the mechanical and biological performance of PLA/PLGA is addressed. In addition, this paper describes the state of the art on challenges and innovations concerning CaP dispersion, incorporation of biomolecules/stem cells and long-term degradation of the composites. PMID:22342596

  1. Differential regulation of Ca2+/calmodulin-dependent enzymes by plant calmodulin isoforms and free Ca2+ concentration.

    PubMed

    Lee, S H; Johnson, J D; Walsh, M P; Van Lierop, J E; Sutherland, C; Xu, A; Snedden, W A; Kosk-Kosicka, D; Fromm, H; Narayanan, N; Cho, M J

    2000-08-15

    Multiple calmodulin (CaM) isoforms are expressed in plants, but their biochemical characteristics are not well resolved. Here we show the differential regulation exhibited by two soya bean CaM isoforms (SCaM-1 and SCaM-4) for the activation of five CaM-dependent enzymes, and the Ca(2+) dependence of their target enzyme activation. SCaM-1 activated myosin light-chain kinase as effectively as brain CaM (K(act) 1.8 and 1.7 nM respectively), but SCaM-4 produced no activation of this enzyme. Both CaM isoforms supported near maximal activation of CaM-dependent protein kinase II (CaM KII), but SCaM-4 exhibited approx.12-fold higher K(act) than SCaM-1 for CaM KII phosphorylation of caldesmon. The SCaM isoforms showed differential activation of plant and animal Ca(2+)-ATPases. The plant Ca(2+)-ATPase was activated maximally by both isoforms, while the erythrocyte Ca(2+)-ATPase was activated only by SCaM-1. Plant glutamate decarboxylase was activated fully by SCaM-1, but SCaM-4 exhibited an approx. 4-fold increase in K(act) and an approx. 25% reduction in V(max). Importantly, SCaM isoforms showed a distinct Ca(2+) concentration requirement for target enzyme activation. SCaM-4 required 4-fold higher [Ca(2+)] for half-maximal activation of CaM KII, and 1.5-fold higher [Ca(2+)] for activation of cyclic nucleotide phosphodiesterase than SCaM-1. Thus these plant CaM isoforms provide a mechanism by which a different subset of target enzymes could be activated or inhibited by the differential expression of these CaM isoforms or by differences in Ca(2+) transients.

  2. Ovarian Hormone Loss Impairs Excitatory Synaptic Transmission at Hippocampal CA3–CA1 Synapses

    PubMed Central

    Bryant, Damani N.; Dorsa, Daniel M.; Adelman, John P.; Maylie, James

    2013-01-01

    Premature and long-term ovarian hormone loss following ovariectomy (OVX) is associated with cognitive impairment. This condition is prevented by estradiol (E2) therapy when initiated shortly following OVX but not after substantial delay. To determine whether these clinical findings are correlated with changes in synaptic functions, we used adult OVX rats to evaluate the consequences of short-term (7–10 d, OVXControl) and long-term (∼5 months, OVXLT) ovarian hormone loss, as well as subsequent in vivo E2 treatment, on excitatory synaptic transmission at the hippocampal CA3–CA1 synapses important for learning and memory. The results show that ovarian hormone loss was associated with a marked decrease in synaptic strength. E2 treatment increased synaptic strength in OVXControl but not OVXLT rats, demonstrating a change in the efficacy for E2 5 months following OVX. E2 also had a more rapid effect: within minutes of bath application, E2 acutely increased synaptic strength in all groups except OVXLT rats that did not receive in vivo E2 treatment. E2's acute effect was mediated postsynaptically, and required Ca2+ influx through the voltage-gated Ca2+ channels. Despite E2's acute effect, synaptic strength of OVXLT rats remained significantly lower than that of OVXControl rats. Thus, changes in CA3–CA1 synaptic transmission associated with ovarian hormone loss cannot be fully reversed with delayed E2 treatment. Given that synaptic strength at CA3–CA1 synapses is related to the ability to learn hippocampus-dependent tasks, these findings provide additional insights for understanding cognitive impairment-associated long-term ovarian hormone loss and ineffectiveness for delayed E2 treatment to maintain cognitive functions. PMID:24107948

  3. Dual Effect of Phosphate Transport on Mitochondrial Ca2+ Dynamics*

    PubMed Central

    Wei, An-Chi; Liu, Ting; O'Rourke, Brian

    2015-01-01

    The large inner membrane electrochemical driving force and restricted volume of the matrix confer unique constraints on mitochondrial ion transport. Cation uptake along with anion and water movement induces swelling if not compensated by other processes. For mitochondrial Ca2+ uptake, these include activation of countertransporters (Na+/Ca2+ exchanger and Na+/H+ exchanger) coupled to the proton gradient, ultimately maintained by the proton pumps of the respiratory chain, and Ca2+ binding to matrix buffers. Inorganic phosphate (Pi) is known to affect both the Ca2+ uptake rate and the buffering reaction, but the role of anion transport in determining mitochondrial Ca2+ dynamics is poorly understood. Here we simultaneously monitor extra- and intra-mitochondrial Ca2+ and mitochondrial membrane potential (ΔΨm) to examine the effects of anion transport on mitochondrial Ca2+ flux and buffering in Pi-depleted guinea pig cardiac mitochondria. Mitochondrial Ca2+ uptake proceeded slowly in the absence of Pi but matrix free Ca2+ ([Ca2+]mito) still rose to ∼50 μm. Pi (0.001–1 mm) accelerated Ca2+ uptake but decreased [Ca2+]mito by almost 50% while restoring ΔΨm. Pi-dependent effects on Ca2+ were blocked by inhibiting the phosphate carrier. Mitochondrial Ca2+ uptake rate was also increased by vanadate (Vi), acetate, ATP, or a non-hydrolyzable ATP analog (AMP-PNP), with differential effects on matrix Ca2+ buffering and ΔΨm recovery. Interestingly, ATP or AMP-PNP prevented the effects of Pi on Ca2+ uptake. The results show that anion transport imposes an upper limit on mitochondrial Ca2+ uptake and modifies the [Ca2+]mito response in a complex manner. PMID:25963147

  4. Dual Effect of Phosphate Transport on Mitochondrial Ca2+ Dynamics.

    PubMed

    Wei, An-Chi; Liu, Ting; O'Rourke, Brian

    2015-06-26

    The large inner membrane electrochemical driving force and restricted volume of the matrix confer unique constraints on mitochondrial ion transport. Cation uptake along with anion and water movement induces swelling if not compensated by other processes. For mitochondrial Ca(2+) uptake, these include activation of countertransporters (Na(+)/Ca(2+) exchanger and Na(+)/H(+) exchanger) coupled to the proton gradient, ultimately maintained by the proton pumps of the respiratory chain, and Ca(2+) binding to matrix buffers. Inorganic phosphate (Pi) is known to affect both the Ca(2+) uptake rate and the buffering reaction, but the role of anion transport in determining mitochondrial Ca(2+) dynamics is poorly understood. Here we simultaneously monitor extra- and intra-mitochondrial Ca(2+) and mitochondrial membrane potential (ΔΨm) to examine the effects of anion transport on mitochondrial Ca(2+) flux and buffering in Pi-depleted guinea pig cardiac mitochondria. Mitochondrial Ca(2+) uptake proceeded slowly in the absence of Pi but matrix free Ca(2+) ([Ca(2+)]mito) still rose to ~50 μm. Pi (0.001-1 mm) accelerated Ca(2+) uptake but decreased [Ca(2+)]mito by almost 50% while restoring ΔΨm. Pi-dependent effects on Ca(2+) were blocked by inhibiting the phosphate carrier. Mitochondrial Ca(2+) uptake rate was also increased by vanadate (Vi), acetate, ATP, or a non-hydrolyzable ATP analog (AMP-PNP), with differential effects on matrix Ca(2+) buffering and ΔΨm recovery. Interestingly, ATP or AMP-PNP prevented the effects of Pi on Ca(2+) uptake. The results show that anion transport imposes an upper limit on mitochondrial Ca(2+) uptake and modifies the [Ca(2+)]mito response in a complex manner. PMID:25963147

  5. Ca-α1T, a fly T-type Ca2+ channel, negatively modulates sleep

    PubMed Central

    Jeong, Kyunghwa; Lee, Soyoung; Seo, Haengsoo; Oh, Yangkyun; Jang, Donghoon; Choe, Joonho; Kim, Daesoo; Lee, Jung-Ha; Jones, Walton D.

    2015-01-01

    Mammalian T-type Ca2+ channels are encoded by three separate genes (Cav3.1, 3.2, 3.3). These channels are reported to be sleep stabilizers important in the generation of the delta rhythms of deep sleep, but controversy remains. The identification of precise physiological functions for the T-type channels has been hindered, at least in part, by the potential for compensation between the products of these three genes and a lack of specific pharmacological inhibitors. Invertebrates have only one T-type channel gene, but its functions are even less well-studied. We cloned Ca-α1T, the only Cav3 channel gene in Drosophila melanogaster, expressed it in Xenopus oocytes and HEK-293 cells, and confirmed it passes typical T-type currents. Voltage-clamp analysis revealed the biophysical properties of Ca-α1T show mixed similarity, sometimes falling closer to Cav3.1, sometimes to Cav3.2, and sometimes to Cav3.3. We found Ca-α1T is broadly expressed across the adult fly brain in a pattern vaguely reminiscent of mammalian T-type channels. In addition, flies lacking Ca-α1T show an abnormal increase in sleep duration most pronounced during subjective day under continuous dark conditions despite normal oscillations of the circadian clock. Thus, our study suggests invertebrate T-type Ca2+ channels promote wakefulness rather than stabilizing sleep. PMID:26647714

  6. Uptake of Ca2+ mediated by the (Ca2+ + Mg2+)-ATPase in reconstituted vesicles.

    PubMed

    Gould, G W; McWhirter, J M; East, J M; Lee, A G

    1987-11-01

    The (Ca2+ + Mg2+)-ATPase was purified from skeletal muscle sarcoplasmic reticulum and reconstituted into sealed phospholipid vesicles by solution in cholate and deoxycholate followed by detergent removal on a column of Sephadex G-50. The level of Ca2+ accumulated by these vesicles, either in the presence or absence of phosphate within the vesicles, increased with increasing content of phosphatidylethanolamine in the phospholipid mixture used for the reconstitution. The levels of Ca2+ accumulated in the absence of phosphate were very low for vesicles reconstituted with egg yolk phosphatidylcholine alone at pH 7.4, but increased markedly with decreasing pH to 6.0. Uptake was also relatively low for vesicles reconstituted with dimyristoleoyl- or dinervonylphosphatidylcholine, and addition of cholesterol had little effect. The level of Ca2+ accumulated increased with increasing external K+ concentration, and was also increased by the ionophores FCCP and valinomycin. Vesicle sizes changed little with changing phosphatidylethanolamine content, and the sidedness of insertion of the ATPase was close to random at all phosphatidylethanolamine contents. It is suggested that the effect of phosphatidylethanolamine on the level of Ca2+ accumulation follows from an effect on the rate of Ca2+ efflux mediated by the ATPase.

  7. Rediscovering area CA2: unique properties and functions

    PubMed Central

    Dudek, Serena M.; Alexander, Georgia M.; Farris, Shannon

    2016-01-01

    Hippocampal area CA2 has several features that distinguish it from CA1 and CA3, including a unique gene expression profile, failure to display long-term potentiation and relative resistance to cell death. A recent increase in interest in the CA2 region, combined with the development of new methods to define and manipulate its neurons, has led to some exciting new discoveries on the properties of CA2 neurons and their role in behaviour. Here, we review these findings and call attention to the idea that the definition of area CA2 ought to be revised in light of gene expression data. PMID:26806628

  8. Endo-lysosomal TRP mucolipin-1 channels trigger global ER Ca2+ release and Ca2+ influx

    PubMed Central

    Kilpatrick, Bethan S.; Yates, Elizabeth; Grimm, Christian; Schapira, Anthony H.

    2016-01-01

    ABSTRACT Transient receptor potential (TRP) mucolipins (TRPMLs), encoded by the MCOLN genes, are patho-physiologically relevant endo-lysosomal ion channels crucial for membrane trafficking. Several lines of evidence suggest that TRPMLs mediate localised Ca2+ release but their role in Ca2+ signalling is not clear. Here, we show that activation of endogenous and recombinant TRPMLs with synthetic agonists evoked global Ca2+ signals in human cells. These signals were blocked by a dominant-negative TRPML1 construct and a TRPML antagonist. We further show that, despite a predominant lysosomal localisation, TRPML1 supports both Ca2+ release and Ca2+ entry. Ca2+ release required lysosomal and ER Ca2+ stores suggesting that TRPMLs, like other endo-lysosomal Ca2+ channels, are capable of ‘chatter’ with ER Ca2+ channels. Our data identify new modalities for TRPML1 action. PMID:27577094

  9. Kinetics and stoichiometry of coupled Na efflux and Ca influx (Na/Ca exchange) in barnacle muscle cells

    PubMed Central

    1989-01-01

    Coupled Na+ exit/Ca2+ entry (Na/Ca exchange operating in the Ca2+ influx mode) was studied in giant barnacle muscle cells by measuring 22Na+ efflux and 45Ca2+ influx in internally perfused, ATP-fueled cells in which the Na+ pump was poisoned by 0.1 mM ouabain. Internal free Ca2+, [Ca2+]i, was controlled with a Ca-EGTA buffering system containing 8 mM EGTA and varying amounts of Ca2+. Ca2+ sequestration in internal stores was inhibited with caffeine and a mitochondrial uncoupler (FCCP). To maximize conditions for Ca2+ influx mode Na/Ca exchange, and to eliminate tracer Na/Na exchange, all of the external Na+ in the standard Na+ sea water (NaSW) was replaced by Tris or Li+ (Tris-SW or LiSW, respectively). In both Na-free solutions an external Ca2+ (Cao)-dependent Na+ efflux was observed when [Ca2+]i was increased above 10(-8) M; this efflux was half-maximally activated by [Ca2+]i = 0.3 microM (LiSW) to 0.7 microM (Tris-SW). The Cao-dependent Na+ efflux was half-maximally activated by [Ca2+]o = 2.0 mM in LiSW and 7.2 mM in Tris-SW; at saturating [Ca2+]o, [Ca2+]i, and [Na+]i the maximal (calculated) Cao-dependent Na+ efflux was approximately 75 pmol#cm2.s. This efflux was inhibited by external Na+ and La3+ with IC50's of approximately 125 and 0.4 mM, respectively. A Nai-dependent Ca2+ influx was also observed in Tris-SW. This Ca2+ influx also required [Ca2+]i greater than 10(-8) M. Internal Ca2+ activated a Nai-independent Ca2+ influx from LiSW (tracer Ca/Ca exchange), but in Tris-SW virtually all of the Cai-activated Ca2+ influx was Nai-dependent (Na/Ca exchange). Half-maximal activation was observed with [Na+]i = 30 mM. The fact that internal Ca2+ activates both a Cao-dependent Na+ efflux and a Nai- dependent Ca2+ influx in Tris-SW implies that these two fluxes are coupled; the activating (intracellular) Ca2+ does not appear to be transported by the exchanger. The maximal (calculated) Nai-dependent Ca2+ influx was -25 pmol/cm2.s. At various [Na+]i between 6 and 106 m

  10. Neuronal Ca(2+) dyshomeostasis in Huntington disease.

    PubMed

    Giacomello, Marta; Oliveros, Juan C; Naranjo, Jose R; Carafoli, Ernesto

    2013-01-01

    The expansion of the N-terminal poly-glutamine tract of the huntingtin (Htt) protein is responsible for Huntington disease (HD). A large number of studies have explored the neuronal phenotype of HD, but the molecular aethiology of the disease is still very poorly understood. This has hampered the development of an appropriate therapeutical strategy to at least alleviate its symptoms. In this short review, we have focused our attention on the alteration of a specific cellular mechanism common to all HD models, either genetic or induced by treatment with 3-NPA, i.e. the cellular dyshomeostasis of Ca(2+). We have highlighted the direct and indirect (i.e. transcriptionally mediated) effects of mutated Htt on the maintenance of the intracellular Ca(2+) balance, the correct modulation of which is fundamental to cell survival and the disturbance of which plays a key role in the death of the cell.

  11. Determination, by using GPR, of the volumetric water content in structures, sub-structures, foundations and soil - ongoing activities in Working Project 2.5 of COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Tosti, Fabio; Slob, Evert

    2015-04-01

    This work will endeavour to review the current status of research activities carried out in Working Project 2.5 'Determination, by using GPR, of the volumetric water content in structures, sub-structures, foundations and soil' within the framework of Working Group 2 'GPR surveying of pavements, bridges, tunnels and buildings; underground utility and void sensing' of the COST (European COoperation in Science and Technology) Action TU1208 'Civil Engineering Applications of Ground Penetrating Radar' (www.GPRadar.eu). Overall, the Project includes 55 Participants from over 21 countries representing 33 Institutions. By considering the type of Institution, a percentage of 64% (35 units) comes from the academic world, while Research Centres and Companies include, respectively, the 27% (15 units) and 9% (5 units) of Institutions. Geographically speaking, Europe is the continent most represented with 18 out of 21 countries, followed by Africa (2 countries) and Asia (1 country). In more details and according to the Europe sub-regions classification provided by the United Nations, Southern Europe includes 39% of countries, Western Europe 27%, while Northern and Eastern Europe are equally present with 17% of countries each. Relying on the main purpose of Working Project 2.5, namely, the ground-penetrating radar-based evaluation of volumetric water content in structures, substructures , foundations, and soils, four main issues have been overall addressed over the first two years of activities. The first one, has been related to provide a comprehensive state of the art on the topic, due to the wide-ranging applications covered in the main disciplines of civil engineering, differently demanding. In this regard, two main publications reviewing the state of the art have been produced [1,2]. Secondly, discussions among Working Group Chairs and other Working Project Leaders have been undertaken and encouraged to avoid the risk of overlapping amongst similar topics from other Working

  12. Luminal Ca2+ promoting spontaneous Ca2+ release from inositol trisphosphate-sensitive stores in rat hepatocytes.

    PubMed Central

    Missiaen, L; Taylor, C W; Berridge, M J

    1992-01-01

    1. Spontaneous Ca2+ release from the inositol 1,4,5-trisphosphate (InsP3)-sensitive stores in permeabilized hepatocytes was monitored using Fluo-3 to measure the free [Ca2+] of the medium bathing the cells. 2. Permeabilized cells rapidly sequestered Ca2+, reducing the [Ca2+] to 103 +/- 5 nM. Under conditions that depended critically upon cell density and the amount of Ca2+ in the medium, this was followed by a slow increase in [Ca2+] culminating in a substantial Ca2+ spike representing synchronous discharge from the InsP3-sensitive stores. 3. During the latency preceding the Ca2+ spike, the stores increased their sensitivity to InsP3. This sensitization seemed to be an all-or-none phenomenon. 4. Oxidized glutathione and thimerosal promoted the spontaneous release by sensitizing the InsP3 receptor. 5. An increase in the [Ca2+] within the stores was required for both the increased sensitivity to InsP3 and the subsequent spike. 6. Caffeine (6 mM) antagonized the effect of very low InsP3 concentrations and abolished the Ca2+ spike, without itself releasing Ca2+. 7. Our results suggesting that luminal Ca2+ may sensitive InsP3-sensitive stores leading to spontaneous Ca2+ mobilization will be discussed in the light of a modified version of the two-pool model for explaining cytosolic Ca2+ oscillations. PMID:1484365

  13. TU-C-12A-09: Modeling Pathologic Response of Locally Advanced Esophageal Cancer to Chemo-Radiotherapy Using Quantitative PET/CT Features, Clinical Parameters and Demographics

    SciTech Connect

    Zhang, H; Chen, W; Kligerman, S; D’Souza, W; Suntharalingam, M; Lu, W; Tan, S; Kim, G

    2014-06-15

    including quantitative PET/CT features accurately and precisely predicted pathologic tumor response to CRT in esophageal cancer. This work was supported in part by National Cancer Institute Grant R21 CA131979 and R01 CA172638. Shan Tan was supported in part by the National Natural Science Foundation of China 60971112 and 61375018, and by Fundamental Research Funds for the Central Universities 2012QN086.

  14. Evidence that Ca2+-release-activated Ca2+ channels in rat hepatocytes are required for the maintenance of hormone-induced Ca2+ oscillations.

    PubMed Central

    Gregory, Roland B; Barritt, Gregory J

    2003-01-01

    Store-operated Ca(2+) channels in liver cells have been shown previously to exhibit a high selectivity for Ca(2+) and to have properties indistinguishable from those of Ca(2+)-release-activated Ca(2+) (CRAC) channels in mast cells and lymphocytes [Rychkov, Brereton, Harland and Barritt (2001) Hepatology 33, 938-947]. The role of CRAC channels in the maintenance of hormone-induced oscillations in the cytoplasmic free Ca(2+) concentration ([Ca(2+)](cyt)) in isolated rat hepatocytes was investigated using several inhibitors of CRAC channels. 2-Aminoethyl diphenylborate (2-APB; 75 microM), Gd(3+) (1 microM) and 1-[beta-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl]-1H-imidazole hydrochloride (SK&F 96365; 50 microM) each inhibited vasopressin- and adrenaline (epinephrine)-induced Ca(2+) oscillations (measured using fura-2). The characteristics of this inhibition were similar to those of inhibition caused by decreasing the extracellular Ca(2+) concentration to zero by addition of EGTA. The effect of 2-APB was reversible. In contrast, LOE-908 [( R, S )-(3,4-dihydro-6,7-dimethoxy-isochinolin-1-yl)-2-phenyl- N, N -di[2-(2,3,4-trimethoxyphenyl)ethyl]acetamide mesylate] (30 microM), used commonly to block Ca(2+) inflow through intracellular-messenger-activated, non-selective cation channels, did not inhibit the Ca(2+) oscillations. In the absence of added extracellular Ca(2+), 2-APB, Gd(3+) and SK&F 96365 did not alter the kinetics of the increase in [Ca(2+)](cyt) induced by a concentration of adrenaline or vasopressin that induces continuous Ca(2+) oscillations at the physiological extracellular Ca(2+) concentration. Ca(2+) inflow through non-selective cation channels activated by maitotoxin could not restore Ca(2+) oscillations in cells treated with 2-APB to block Ca(2+) inflow through CRAC channels. Evidence for the specificity of the pharmacological agents for inhibition of CRAC channels under the conditions of the present experiments with hepatocytes is discussed. It

  15. Superconductivity in Ca-doped graphene laminates.

    PubMed

    Chapman, J; Su, Y; Howard, C A; Kundys, D; Grigorenko, A N; Guinea, F; Geim, A K; Grigorieva, I V; Nair, R R

    2016-03-16

    Despite graphene's long list of exceptional electronic properties and many theoretical predictions regarding the possibility of superconductivity in graphene, its direct and unambiguous experimental observation has not been achieved. We searched for superconductivity in weakly interacting, metal decorated graphene crystals assembled into so-called graphene laminates, consisting of well separated and electronically decoupled graphene crystallites. We report robust superconductivity in all Ca-doped graphene laminates. They become superconducting at temperatures (Tc) between ≈4 and ≈6 K, with Tc's strongly dependent on the confinement of the Ca layer and the induced charge carrier concentration in graphene. We find that Ca is the only dopant that induces superconductivity in graphene laminates above 1.8 K among several dopants used in our experiments, such as potassium, caesium and lithium. By revealing the tunability of the superconducting response through doping and confinement of the metal layer, our work shows that achieving superconductivity in free-standing, metal decorated monolayer graphene is conditional on an optimum confinement of the metal layer and sufficient doping, thereby bringing its experimental realization within grasp.

  16. Positive magnetoresistance in Ca-doped cobaltites

    SciTech Connect

    Zhou, S. M. Li, Y.; Guo, Y. Q.; Zhao, J. Y.; Shi, L.

    2014-12-08

    Transport properties of polycrystalline La{sub 1−x}Ca{sub x}CoO{sub 3} (0.10 ≤ x ≤ 0.25) are systemically studied in this work. Three types of magnetoresistance (MR) effects are found in the Ca-doped cobaltites. Two negative MRs appear around high-temperature ferromagnetic transition and at low temperatures, which correspond to the conventional MR due to the field-induced suppression of spin-disorder scattering and the intergranular giant-MR due to spin-dependent transport between the ferromagnetic clusters, respectively. More interestingly, another exotic positive MR emerges at intermediate temperature region, which had not been previously reported in Sr- and Ba-doped cobaltites. It is found that this positive MR is associated with an abnormally magnetic transition and increases with the increase of x. For x = 0.25, the MR at low temperatures is dominated by the positive one, which is isotropic and nearly linear with the magnetic field. The possible origin of the positive MR in the Ca-doped cobaltites is discussed.

  17. Superconductivity in CaBi2.

    PubMed

    Winiarski, M J; Wiendlocha, B; Gołąb, S; Kushwaha, S K; Wiśniewski, P; Kaczorowski, D; Thompson, J D; Cava, R J; Klimczuk, T

    2016-08-01

    Superconductivity is observed with critical temperature Tc = 2.0 K in self-flux-grown single crystals of CaBi2. This material adopts the ZrSi2 structure type with lattice parameters a = 4.696(1) Å, b = 17.081(2) Å and c = 4.611(1) Å. The crystals of CaBi2 were studied by means of magnetic susceptibility, specific heat and electrical resistivity measurements. The heat capacity jump at Tc is ΔC/γTc = 1.41, confirming bulk superconductivity; the Sommerfeld coefficient γ = 4.1 mJ mol(-1) K(-2) and the Debye temperature ΘD = 157 K. The electron-phonon coupling strength is λel-ph = 0.59, and the thermodynamic critical field Hc is low, between 111 and 124 Oe CaBi2 is a moderate coupling type-I superconductor. Results of electronic structure calculations are reported and charge densities, electronic bands, densities of states and Fermi surfaces are discussed, focusing on the effects of spin-orbit coupling and electronic property anisotropy. We find a mixed quasi-2D + 3D character in the electronic structure, which reflects the layered crystal structure of the material. PMID:27435423

  18. Superconductivity in CaBi2.

    PubMed

    Winiarski, M J; Wiendlocha, B; Gołąb, S; Kushwaha, S K; Wiśniewski, P; Kaczorowski, D; Thompson, J D; Cava, R J; Klimczuk, T

    2016-08-01

    Superconductivity is observed with critical temperature Tc = 2.0 K in self-flux-grown single crystals of CaBi2. This material adopts the ZrSi2 structure type with lattice parameters a = 4.696(1) Å, b = 17.081(2) Å and c = 4.611(1) Å. The crystals of CaBi2 were studied by means of magnetic susceptibility, specific heat and electrical resistivity measurements. The heat capacity jump at Tc is ΔC/γTc = 1.41, confirming bulk superconductivity; the Sommerfeld coefficient γ = 4.1 mJ mol(-1) K(-2) and the Debye temperature ΘD = 157 K. The electron-phonon coupling strength is λel-ph = 0.59, and the thermodynamic critical field Hc is low, between 111 and 124 Oe CaBi2 is a moderate coupling type-I superconductor. Results of electronic structure calculations are reported and charge densities, electronic bands, densities of states and Fermi surfaces are discussed, focusing on the effects of spin-orbit coupling and electronic property anisotropy. We find a mixed quasi-2D + 3D character in the electronic structure, which reflects the layered crystal structure of the material.

  19. Toward Laser Cooling of CaF

    NASA Astrophysics Data System (ADS)

    Hemmerling, Boerge; Drayna, Garrett; Chae, Eunmi; Ravi, Aakash; Lu, Hsin-I.; Yeo, Mark; Hummon, Matthew; Collopy, Alejandra; Stuhl, Benjamin; Ye, Jun; Doyle, John

    2013-05-01

    The prospects of novel physics employing polar cold molecules encompass quantum computing and simulations, controlled ultra-cold chemistry and precision measurements. However, a method liable to bring a general class of chemically diverse molecules to the ultracold regime still needs to be developed. We report on the progress of experiments to laser cool CaF molecules, including the implementation of a magneto-optical trap (MOT). We use a 2-stage buffer-gas cooled beam source to produce a cold and slow beam of particles. In this experiment, we plan to load the trap from this buffer-gas source. As a precursor to working with CaF, we successfully implemented the first buffer-gas loaded MOT of Yb, without the use of a Zeeman slower, but using only a non-chirped slowing laser. The lifetime of the MOT was measured to be > 100 ms, with the distance between the source and the MOT ~ 30 cm. We describe a scheme for the laser cooling and magneto-optical confinement of CaF molecules, following an approach similar to those used in the cooling of SrF and YO.

  20. Superconductivity in Ca-doped graphene laminates

    PubMed Central

    Chapman, J.; Su, Y.; Howard, C. A.; Kundys, D.; Grigorenko, A. N.; Guinea, F.; Geim, A. K.; Grigorieva, I. V.; Nair, R. R.

    2016-01-01

    Despite graphene’s long list of exceptional electronic properties and many theoretical predictions regarding the possibility of superconductivity in graphene, its direct and unambiguous experimental observation has not been achieved. We searched for superconductivity in weakly interacting, metal decorated graphene crystals assembled into so-called graphene laminates, consisting of well separated and electronically decoupled graphene crystallites. We report robust superconductivity in all Ca-doped graphene laminates. They become superconducting at temperatures (Tc) between ≈4 and ≈6 K, with Tc’s strongly dependent on the confinement of the Ca layer and the induced charge carrier concentration in graphene. We find that Ca is the only dopant that induces superconductivity in graphene laminates above 1.8 K among several dopants used in our experiments, such as potassium, caesium and lithium. By revealing the tunability of the superconducting response through doping and confinement of the metal layer, our work shows that achieving superconductivity in free-standing, metal decorated monolayer graphene is conditional on an optimum confinement of the metal layer and sufficient doping, thereby bringing its experimental realization within grasp. PMID:26979564

  1. Ca sup + emission in the sunlit ionosphere

    SciTech Connect

    Torr, M.R. ); Torr, D.G.; Bhatt, P.; Swift, W.; Dougani, H. )

    1990-03-01

    In the course of a program of twilight airglow observations from the McDonald Observatory in southwest Texas, the resonance fluorescence emissions from calcium ions were measured. In particular, twilight sequences were obtained during the period of December 19-22, 1987, which coincided with the Ursids meteorite shower. During this meteorite event the intensities of the Ca{sup +} emission lines at 3,934 {angstrom} increased to the point that the surface brightness profiles could be inverted to volume emission rate profiles. These profiles show evidence for strong spatial redistribution of the Ca{sup +} over the course of three days. Prior to the onset of the meteorite activity, emissions from the Ca{sup +} originate from below 100 km, on the occasions when the emissions are visible. By the evening of December 19 a peak is measurable at 108 km. On the morning of December 22, a high-altitude peak was observed above 250 km, with a larger peak down at approximately 85 km. By the evening of December 22, the emission had substantially intensified, with the peak of the layer being at 80 km or below, but with emission being produced all the way up to at least 160 km. Observations of these emissions during meteor shower periods could provide a valuable tracer for the processes responsible for the transport of ions in the D, E, and F region, allowing the full altitude and latitude extent of the distribution to be determined.

  2. Superconductivity in Ca-doped graphene laminates

    NASA Astrophysics Data System (ADS)

    Chapman, J.; Su, Y.; Howard, C. A.; Kundys, D.; Grigorenko, A. N.; Guinea, F.; Geim, A. K.; Grigorieva, I. V.; Nair, R. R.

    2016-03-01

    Despite graphene’s long list of exceptional electronic properties and many theoretical predictions regarding the possibility of superconductivity in graphene, its direct and unambiguous experimental observation has not been achieved. We searched for superconductivity in weakly interacting, metal decorated graphene crystals assembled into so-called graphene laminates, consisting of well separated and electronically decoupled graphene crystallites. We report robust superconductivity in all Ca-doped graphene laminates. They become superconducting at temperatures (Tc) between ≈4 and ≈6 K, with Tc’s strongly dependent on the confinement of the Ca layer and the induced charge carrier concentration in graphene. We find that Ca is the only dopant that induces superconductivity in graphene laminates above 1.8 K among several dopants used in our experiments, such as potassium, caesium and lithium. By revealing the tunability of the superconducting response through doping and confinement of the metal layer, our work shows that achieving superconductivity in free-standing, metal decorated monolayer graphene is conditional on an optimum confinement of the metal layer and sufficient doping, thereby bringing its experimental realization within grasp.

  3. CA 19-9: handle with care.

    PubMed

    Galli, Claudio; Basso, Daniela; Plebani, Mario

    2013-07-01

    Since its inception in the mid-1980s of the 20th century testing for carbohydrate antigen 19-9 (CA 19-9) has raised expectation for an earlier diagnosis and accurate monitoring of several malignant diseases. After almost 30 years, the available evidences have confirmed the appropriateness and usefulness of determining CA 19-9 levels as a prognostic indicator and as a reliable tool for monitoring pancreatic and gastrointestinal cancer, but concerns have been raised about its applications in screening, which is actually not recommended, and in the diagnosis of malignancies, due to several interferences that limit the specificity and to the insufficient sensitivity of this marker. In this paper we aimed to review the basic concepts of CA 19-9 testing and its current applications, with a major focus on the most recent evidences dealing with assay interference, methods comparison and monitoring of malignant diseases. The prognostic value and monitoring recommendations for pancreatic, gastric and colorectal cancers are described in depth.

  4. Comparison of Ca and Ar Diffusion in Phlogopite: Implications for K-Ca and K-Ar Geochronology

    NASA Astrophysics Data System (ADS)

    Cruz, M. F.; Szilas, K.; Grove, M. J.

    2015-12-01

    Coupled geochronology based upon branched decay of 40K-40Ar and 40K-40Ca decay is rarely exploited because 40Ca is the major common isotope of calcium and 40Ca and 40K are difficult to resolve isotopically without resorting to isotope dilution wet chemistry. Recently developed ion microprobe methods based upon measurement of doubly ionized species partially overcome the latter problem and have been applied to high K/Ca micas. The ability to interpret K-Ar and K-Ca results is limited due to uncertainty in the relative diffusion properties of Ca and Ar. To address this problem, we are performing Ar and Ca diffusion experiments and fluid-crystal Ar partitioning experiments with anhydrous F-phlogopite that is stable to 1390°C. As an additional check, we are comparing K-Ca and K-Ar ages from natural mantle phlogopites from a variety of settings to assess the relative retentivity of Ar and Ca. South African xenoliths tend to yield 40Ar/39Ar ages that are much older than K-Ca ages from the same phologopites. Possible excess 40Ar and high common Ca render the comparisons inconclusive, but this suggests that Ca diffuses more readily than Ar in phlogopite. Our most definitive K-Ca phlogopite results (i.e., least affected by common Ca) come from the Archean Seqi dunite of SW Greenland. The K-Ca ages of Seqi phlogopites is 927 ± 26 Ma (2s). Incremental heating 40Ar/39Ar results from the same sample yields a much older result with a terminal age of 3.5 Ga. However, the first 5-10% of 39Ar release are consistent with transient heating at ca. 900 Ma. Considered together, the K-Ca and 40Ar/39Ar results from the Seqi dunite locality strongly suggest that Ca diffusion is more rapid than Ar diffusion in phlogopite.

  5. Dissociation of Ca-bearing Molecules as a Source of Mercury's Ca Exosphere

    NASA Astrophysics Data System (ADS)

    Burger, Matthew H.; Killen, Rosemary M.

    2015-11-01

    Observations of Mercury's calcium exosphere by MESSENGER have revealed three key features: (1) The Ca is extremely energetic, with a temperature ~70,000 K if the source is thermal, (2) the source region is located in the dawn hemisphere, and (3) there is a strong annual variation in the Ca source rate (Burger et al. 2014). Killen and Hahn (2015) have shown that the source rate is consistent with impact vaporization by interplanetary dust and the intersection of Mercury with a cometary dust stream (likely associated with Comet Encke, Christou et al., submitted).Killen et al. (2005) suggested that energetic calcium could be produced by the dissociation of Ca-bearing molecules produced in impact vaporization plumes. We test this hypothesis with a Monte Carlo model that follows the evolution of atomic and molecular calcium produced in impact plumes. Ca-bearing molecules such as CaO, CaOH, and Ca(OH)2 are more likely to be are produced in vapor plumes than atomic Ca (Berezhnoy and Klumov 2008); these molecules quickly break apart either through vibrational dissociation or photodissociation. The excess energy associated with dissociation gives the atomic Ca an extra energy boost above the temperature of the impact plumes (~5000 K). We determine impact vaporization rates and excess energies required by the dissociation process to reproduce the scale height and spatial morphology of the Ca exosphere as observed by the MESSENGER Ultraviolet and Visible Spectrometer (UVVS).ReferencesBerezhnoy, A.A. and Klumov, B.A., Impacts as sources of the exosphere on Mercury, Icarus, 195, 511-522, 2008, doi:10.1016/j.icarus.2008.01.005.Burger, M.H., et al., Seasonal variations in Mercury's dayside calcium exosphere, Icarus, 238, 51-58, 2014, doi:10.1016/j.icarus.2014.04.049.Killen, R.M., et al., The calcium exosphere of Mercury, Icarus, 173, 300-311, 2005, doi:10.1016/j.icarus.2004.08.022.Killen, R.M. and Hahn, J.M., Impact vaporization as a possible source ofMercury's calcium exosphere

  6. Predicting Ca2+ -binding sites using refined carbon clusters.

    PubMed

    Zhao, Kun; Wang, Xue; Wong, Hing C; Wohlhueter, Robert; Kirberger, Michael P; Chen, Guantao; Yang, Jenny J

    2012-12-01

    Identifying Ca(2+) -binding sites in proteins is the first step toward understanding the molecular basis of diseases related to Ca(2+) -binding proteins. Currently, these sites are identified in structures either through X-ray crystallography or NMR analysis. However, Ca(2+) -binding sites are not always visible in X-ray structures due to flexibility in the binding region or low occupancy in a Ca(2+) -binding site. Similarly, both Ca(2+) and its ligand oxygens are not directly observed in NMR structures. To improve our ability to predict Ca(2+) -binding sites in both X-ray and NMR structures, we report a new graph theory algorithm (MUG(C) ) to predict Ca(2+) -binding sites. Using carbon atoms covalently bonded to the chelating oxygen atoms, and without explicit reference to side-chain oxygen ligand co-ordinates, MUG(C) is able to achieve 94% sensitivity with 76% selectivity on a dataset of X-ray structures composed of 43 Ca(2+) -binding proteins. Additionally, prediction of Ca(2+) -binding sites in NMR structures was obtained by MUG(C) using a different set of parameters, which were determined by the analysis of both Ca(2+) -constrained and unconstrained Ca(2+) -loaded structures derived from NMR data. MUG(C) identified 20 of 21 Ca(2+) -binding sites in NMR structures inferred without the use of Ca(2+) constraints. MUG(C) predictions are also highly selective for Ca(2+) -binding sites as analyses of binding sites for Mg(2+) , Zn(2+) , and Pb(2+) were not identified as Ca(2+) -binding sites. These results indicate that the geometric arrangement of the second-shell carbon cluster is sufficient not only for accurate identification of Ca(2+) -binding sites in NMR and X-ray structures but also for selective differentiation between Ca(2+) and other relevant divalent cations.

  7. Regulation of RYR2 by sarcoplasmic reticulum Ca(2+).

    PubMed

    Zhang, Joe Z; Waddell, Helen M M; Jones, Peter P

    2015-06-01

    Ca(2+) is arguably the most important ion involved in the contraction of the heart. The cardiac ryanodine receptor (RyR2), the major Ca(2+) release channel located in the sarcoplasmic reticulum (SR) membrane, is responsible for releasing the bulk of Ca(2+) required for contraction. Moreover, RyR2 is also crucial for maintaining SR Ca(2+) homeostasis by releasing Ca(2+) from the SR when it becomes overloaded with Ca(2+) . During normal contraction, RyR2 is activated by cytosolic Ca(2+) , whereas during store overload conditions, the opening of RyR2 is governed by SR Ca(2+) . Although the process of the cytosolic control of RyR2 is well established, the molecular mechanism by which SR luminal Ca(2+) regulates RyR2 has only recently been elucidated and remains controversial. In addition to the activation of RyR2, SR luminal Ca(2+) also determines when the RyR2 channel closes. RyR2-mediated Ca(2+) release from the SR does not continue until the SR is completely depleted. Rather, it ceases when SR luminal Ca(2+) falls below a certain level. Given the importance of SR Ca(2+) , it is not surprising that the SR luminal Ca(2+) level is tightly controlled by SR Ca(2+) -buffering proteins. Consequently, the opening and closing of RyR2 is heavily influenced by the presence of such proteins, particularly those associated with RyR2, such as calsequestrin and the histidine-rich Ca(2+) -binding protein. These proteins appear to indirectly alter RyR2 activity by modifying the microdomain SR Ca(2+) level surrounding RyR2. PMID:25603835

  8. Conserved properties of individual Ca2+-binding sites in calmodulin

    PubMed Central

    Halling, D. Brent; Liebeskind, Benjamin J.; Hall, Amelia W.; Aldrich, Richard W.

    2016-01-01

    Calmodulin (CaM) is a Ca2+-sensing protein that is highly conserved and ubiquitous in eukaryotes. In humans it is a locus of life-threatening cardiomyopathies. The primary function of CaM is to transduce Ca2+ concentration into cellular signals by binding to a wide range of target proteins in a Ca2+-dependent manner. We do not fully understand how CaM performs its role as a high-fidelity signal transducer for more than 300 target proteins, but diversity among its four Ca2+-binding sites, called EF-hands, may contribute to CaM’s functional versatility. We therefore looked at the conservation of CaM sequences over deep evolutionary time, focusing primarily on the four EF-hand motifs. Expanding on previous work, we found that CaM evolves slowly but that its evolutionary rate is substantially faster in fungi. We also found that the four EF-hands have distinguishing biophysical and structural properties that span eukaryotes. These results suggest that all eukaryotes require CaM to decode Ca2+ signals using four specialized EF-hands, each with specific, conserved traits. In addition, we provide an extensive map of sites associated with target proteins and with human disease and correlate these with evolutionary sequence diversity. Our comprehensive evolutionary analysis provides a basis for understanding the sequence space associated with CaM function and should help guide future work on the relationship between structure, function, and disease. PMID:26884197

  9. Ca cycling and isotopic fluxes in forested ecosystems in Hawaii

    USGS Publications Warehouse

    Wiegand, B.A.; Chadwick, O.A.; Vitousek, P.M.; Wooden, J.L.

    2005-01-01

    Biogeochemical processes fractionate Ca isotopes in plants and soils along a 4 million year developmental sequence in the Hawaiian Islands. We observed that plants preferentially take up 40Ca relative to 44Ca, and that biological fractionation and changes in the relative contributions from volcanic and marine sources produce a significant increase in 44Ca in soil exchangeable pools. Our results imply moderate fluxes enriched in 44Ca from strongly nutrient-depleted old soils, in contrast with high 40Ca fluxes in young and little weathered environments. In addition, biological fractionation controls divergent geochemical pathways of Ca and Sr in the plant-soil system. While Ca depletes progressively with increasing soil age, Sr/Ca ratios increase systematically. Sr isotope ratios provide a valuable tracer for provenance studies of alkaline earth elements in forested ecosystems, but its usefulness is limited when deciphering biogeochemical processes involved in the terrestrial Ca cycle. Ca isotopes in combination with Sr/ Ca ratios reveal more complex processes involved in the biogeochemistry of Ca and Sr. Copyright 2005 by the American Geophysical Union.

  10. Ca Isotopic Ratios in Igneous Rocks: Some Preliminary Results

    NASA Astrophysics Data System (ADS)

    Huang, S.; Farkas, J.; Jacobsen, S. B.

    2009-12-01

    Calcium (Ca) is the 5th most abundant element on the Earth, and it is an important geochemical and cosmochemical tracer. It has six isotopes and only H and He have a larger percentage mass difference (Δm/m) between the heaviest and the lightest isotopes. Systematic Ca isotopic studies have mostly focused on low-temperature geochemical processes, and most Ca isotopic analyses have been applied on modern and ancient marine carbonates and sulphates, documenting large and systematic isotopic variations, which were used to infer the chemical evolution of seawater. Detailed work on igneous rocks is very limited. Here we show two examples of how stable Ca isotopic ratios can be a useful geochemical tool in understanding igneous processes. Ca isotopic fractionation between coexisting clinopyroxene and orthopyroxene from mantle peridotites: We report Ca isotopic ratios on co-existing clino- and ortho-pyroxenes from Kilbourne Hole and San Carlos mantle peridotites. The 44Ca/40Ca in orthopyroxenes is ~0.5 per mil heavier than that in co-existing clinopyroxenes. Combined with published Ca isotopic data on low-temperature Ca-bearing minerals (calcite, aragonite and barite), we show that the fractionation of Ca isotopes between Ca-bearing minerals (at both low-temperature and high-temperature) is primarily controlled by the strength of Ca-O bond in the minerals. The mineral with shorter (i.e., stronger) Ca-O bond yields heavier Ca isotopic ratio. Using our measured 44Ca/40Ca in mantle pyroxenes and the relative proportions of major Ca-bearing minerals in the upper mantle, the estimated 44Ca/40Ca of the upper mantle is 1.1 per mil heavier relative to the NIST 915a, ~0.1 to 0.2 per mil higher than basalts. Ca isotopic variation in Hawaiian shield lavas: Large geochemical and isotopic variations have been observed in lavas forming the large tholeiitic shields of Hawaiian volcanoes, with lavas from the surface of the Koolau volcano (Makapuu-stage) defining one compositional and

  11. Determination of the calcium species in coal chars by Ca K-edge XANES analysis

    NASA Astrophysics Data System (ADS)

    Liu, Li-Juan; Liu, Hui-Jun; Cui, Ming-Qi; Hu, Yong-Feng; Zheng, Lei; Zhao, Yi-Dong; Ma, Chen-Yan; Xi, Shi-Bo; Yang, Dong-Liang; Guo, Zhi-Ying; Wang, Jie

    2013-02-01

    Ca-based additives have been widely used as a sulfur adsorbent during coal pyrolysis and gasification. The Ca speciation and evolution during the pyrolysis of coal with Ca additives have attracted great attention. In this paper, Ca species in the coal chars prepared from the pyrolysis of Ca(OH)2 or CaCO3-added coals are studied by using Ca K-edge X-ray absorption near-edge structural spectroscopy. The results demonstrate that Ca(OH)2, CaSO4, CaS and CaO coexist in the Ca(OH)2-added chars, while Ca(OH)2 and CaSO4 are the main species in the Ca(OH)2-added chars. Besides, a carboxyl-bound Ca is also formed during both the pyrolysis for the Ca(OH)2-added and the CaCO3-added coals. A detailed discussion about the Ca speciation is given.

  12. Pressure-induced structural transformation of CaC2.

    PubMed

    Wang, Lu; Huang, Xiaoli; Li, Da; Huang, Yanping; Bao, Kuo; Li, Fangfei; Wu, Gang; Liu, Bingbing; Cui, Tian

    2016-05-21

    The high pressure structural changes of calcium carbide CaC2 have been investigated with Raman spectroscopy and synchrotron X-ray diffraction (XRD) techniques in a diamond anvil cell at room temperature. At ambient conditions, two forms of CaC2 co-exist. Above 4.9 GPa, monoclinic CaC2-ii diminished indicating the structural phase transition from CaC2-ii to CaC2-i. At about 7.0 GPa, both XRD patterns and Raman spectra confirmed that CaC2-i transforms into a metallic Cmcm structure which contains polymeric carbon chains. Along with the phase transition, the isolated C2 dumbbells are polymerized into zigzag chains resulting in a large volume collapse with 22.4%. Above 30.0 GPa, the XRD patterns of CaC2 become featureless and remain featureless upon decompression, suggesting an irreversible amorphization of CaC2.

  13. Pressure-induced structural transformation of CaC2

    NASA Astrophysics Data System (ADS)

    Wang, Lu; Huang, Xiaoli; Li, Da; Huang, Yanping; Bao, Kuo; Li, Fangfei; Wu, Gang; Liu, Bingbing; Cui, Tian

    2016-05-01

    The high pressure structural changes of calcium carbide CaC2 have been investigated with Raman spectroscopy and synchrotron X-ray diffraction (XRD) techniques in a diamond anvil cell at room temperature. At ambient conditions, two forms of CaC2 co-exist. Above 4.9 GPa, monoclinic CaC2-ii diminished indicating the structural phase transition from CaC2-ii to CaC2-i. At about 7.0 GPa, both XRD patterns and Raman spectra confirmed that CaC2-i transforms into a metallic Cmcm structure which contains polymeric carbon chains. Along with the phase transition, the isolated C2 dumbbells are polymerized into zigzag chains resulting in a large volume collapse with 22.4%. Above 30.0 GPa, the XRD patterns of CaC2 become featureless and remain featureless upon decompression, suggesting an irreversible amorphization of CaC2.

  14. Ca2+ triggers massive exocytosis in Chinese hamster ovary cells.

    PubMed Central

    Coorssen, J R; Schmitt, H; Almers, W

    1996-01-01

    We have tracked the cell surface area of CHO cells by measuring the membrane capacitance, Cm. An increase in cytosolic [Ca2+], [Ca2+]i, increased the cell surface area by 20-30%. At micromolar [Ca2+]i the increase occurred in minutes, while at 20 microM or higher [Ca2+]i it occurred in seconds and was transient. GTPgammaS caused a 3% increase even at 0.1 microM [Ca2+]i. We conclude that CHO cells, previously thought capable only of constitutive exocytosis, can perform Ca2+-triggered exocytosis that is both massive and rapid. Ca2+-triggered exocytosis was also observed in 3T3 fibroblasts. Our findings add evidence to the view that Ca induces exocytosis in cells other than known secretory cells. PMID:8670883

  15. Phosphate Capacities of CaF2-MgO and CaF2-CaO-MgO Slags

    NASA Astrophysics Data System (ADS)

    Akbari, F.; Pickles, C. A.

    2015-02-01

    Previously published sulphide capacity data and thermodynamic arguments have been employed to calculate the phosphate capacities and the phosphorus partition ratios between a molten carbon saturated iron alloy and binary CaF2-MgO slags and also ternary CaF2 -CaO-MgO slags at 1450 °C. For the CaF2-MgO binary system, a linear relationship was found between the phosphate and the sulphide capacities as follows: log ? = 1.2 log Cs + 25.2. For the ternary CaF2-CaO-MgO system at 1450 °C, the logarithm of the calculated phosphate capacities ranged from 19.47 to 20.15. With the addition of CaO, the phosphate capacities initially increased, reached a maximum and then decreased slightly. The addition of MgO to the CaF2-CaO system resulted in a decrease in the phosphate capacity. The calculated phosphorus partition ratios increased slightly with increasing mole fraction of CaO in the ternary system.

  16. Application of Ca stable isotopes to long-term changes in the Ca cycle of a Northern Hardwood forest

    NASA Astrophysics Data System (ADS)

    Kurtz, A. C.; Takagi, K.; Bailey, S. W.; Bullen, T. D.

    2015-12-01

    The Hubbard Brook Ecosystem Study (New Hampshire, USA) presents an unusual opportunity for the application of innovative isotope methods in forest biogeochemistry. Changes in biogeochemical cycling resulting from decades of acid deposition, subsequent reductions in acid deposition, and a series of experimental