289 research outputs found

    Model Checking: Verification or Debugging?

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    Scheduling Optimisations for SPIN to Minimise Buffer Requirements in Synchronous Data Flow:(with appendix)

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    Synchronous Data flow (SDF) graphs have a simple and elegant semantics (essentially linear algebra) which makes SDF graphs eminently suitable as a vehicle for studying scheduling optimisations. We extend related work on using SPIN to experiment with scheduling optimisations aimed at minimising buffer requirements. We show that for a benchmark of commonly used case studies the performance of our SPIN based scheduler is comparable to that of state of the art research tools. The key to success is using the semantics of SDF to prove when using (even unsound and/or incomplete) optimisations are justified. The main benefit of our approach lies in gaining deep insight in the optimisations at relatively low cost

    Regulation and role of differential ethylene biosynthesis in gravistimulated Antirrhinum majus L. cut flower stems

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    Gravistimulation induced differential ethylene production in Antirrhinum majus L. cut flower stems with highest levels in the lower halves of the gravistimulated stems. Expression levels of three different 1-aminocyclopropane-1-carboxylate (ACC) synthase (ACS) genes, an ACC oxidase (ACO) and an ethylene receptor (ETR/ERS homolog) gene were studied in the bending zone after 9 h of gravistimulation. One of the ACS genes (Am-ACS3) was abundantly expressed in the lower halves but not in the upper halves of gravistimulated stems. This strongly suggests that Am-ACS3 is responsible for the observed differential ethylene production in gravistimulated stems. Am-ACO and Am-ETR/ERS gene expression was increased in both the lower and upper halves of gravistimulated stems, suggesting that they play no role in differential ethylene production. When gravistimulation was performed in an environment enriched with either 20 µL/L ethylene or 100 nL/L 1-methylcyclopropene (1-MCP), a slight stimulation of bending by 1-MCP and a slight inhibition of bending by ethylene were observed. The regulation and role of ethylene in gravitropism of cut snapdragon flowering stems is discussed

    An auxin-responsive 1-aminocyclopropane-1-carboxylate synthase is responsible for differential ethylene production in gravistimulated Antirrhinum majus L. flower stems

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    The regulation of gravistimulation-induced ethylene production and its role in gravitropic bending was studied in Antirrhinum majus L. cut flower stems. Gravistimulation increased ethylene production in both lower and upper halves of the stems with much higher levels observed in the lower half. Expression patterns of three different 1-aminocyclopropane-1-carboxylate (ACC) synthase (ACS) genes, an ACC oxidase (ACO) and an ethylene receptor (ETR/ERS homolog) gene were studied in the bending zone of gravistimulated stems and in excised stem sections following treatment with different chemicals. One of the ACS genes (Am-ACS3) was abundantly expressed in the bending zone cortex at the lower side of the stems within 2 h of gravistimulation. Am-ACS3 was not expressed in vertical stems or in other parts of (gravistimulated) stems, leaves or flowers. Am-ACS3 was strongly induced by indole-3-acetic acid (IAA) but not responsive to ethylene. The Am-ACS3 expression pattern strongly suggests that Am-ACS3 is responsible for the observed differential ethylene production in gravistimulated stems; its responsiveness to IAA suggests that Am-ACS3 expression reflects changes in auxin signalling. Am-ACS1 also showed increased expression in gravistimulated and IAA-treated stems although to a much lesser extent than Am-ACS3. In contrast to Am-ACS3, Am-ACS1 was also expressed in non-bending regions of vertical and gravistimulated stems and in leaves, and Am-ACS1 expression was not confined to the lower side cortex but evenly distributed over the diameter of the stem. Am-ACO and Am-ETR/ERS expression was increased in both the lower and upper halves of gravistimulated stems. Expression of both Am-ACO and Am-ETR/ERS was responsive to ethylene, suggesting regulation by IAA-dependent differential ethylene production. Am-ACO expression and in vivo ACO activity, in addition, were induced by IAA, independent of the IAA-induced ethylene. IAA-induced growth of vertical stem sections and bending of gravistimulated flowering stems were little affected by ethylene or 1-methylcyclopropene treatments, indicating that the differential ethylene production plays no pivotal role in the kinetics of gravitropic bending

    Combination antiretroviral therapy and the risk of myocardial infarction

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    Identification of autophosphorylation sites in eukaryotic elongation factor-2 kinase

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    eEF2K [eEF2 (eukaryotic elongation factor 2) kinase] phosphorylates and inactivates the translation elongation factor eEF2. eEF2K is not a member of the main eukaryotic protein kinase superfamily, but instead belongs to a small group of so-called α-kinases. The activity of eEF2K is normally dependent upon Ca2+ and calmodulin. eEF2K has previously been shown to undergo autophosphorylation, the stoichiometry of which suggested the existence of multiple sites. In the present study we have identified several autophosphorylation sites, including Thr348, Thr353, Ser366 and Ser445, all of which are highly conserved among vertebrate eEF2Ks. We also identified a number of other sites, including Ser78, a known site of phosphorylation, and others, some of which are less well conserved. None of the sites lies in the catalytic domain, but three affect eEF2K activity. Mutation of Ser78, Thr348 and Ser366 to a non-phosphorylatable alanine residue decreased eEF2K activity. Phosphorylation of Thr348 was detected by immunoblotting after transfecting wild-type eEF2K into HEK (human embryonic kidney)-293 cells, but not after transfection with a kinase-inactive construct, confirming that this is indeed a site of autophosphorylation. Thr348 appears to be constitutively autophosphorylated in vitro. Interestingly, other recent data suggest that the corresponding residue in other α-kinases is also autophosphorylated and contributes to the activation of these enzymes [Crawley, Gharaei, Ye, Yang, Raveh, London, Schueler-Furman, Jia and Cote (2011) J. Biol. Chem. 286, 2607–2616]. Ser366 phosphorylation was also detected in intact cells, but was still observed in the kinase-inactive construct, demonstrating that this site is phosphorylated not only autocatalytically but also in trans by other kinases
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