245 research outputs found
An Entropy Stable Nodal Discontinuous Galerkin Method for the Two Dimensional Shallow Water Equations on Unstructured Curvilinear Meshes with Discontinuous Bathymetry
We design an arbitrary high-order accurate nodal discontinuous Galerkin
spectral element approximation for the nonlinear two dimensional shallow water
equations with non-constant, possibly discontinuous, bathymetry on
unstructured, possibly curved, quadrilateral meshes. The scheme is derived from
an equivalent flux differencing formulation of the split form of the equations.
We prove that this discretisation exactly preserves the local mass and
momentum. Furthermore, combined with a special numerical interface flux
function, the method exactly preserves the mathematical entropy, which is the
total energy for the shallow water equations. By adding a specific form of
interface dissipation to the baseline entropy conserving scheme we create a
provably entropy stable scheme. That is, the numerical scheme discretely
satisfies the second law of thermodynamics. Finally, with a particular
discretisation of the bathymetry source term we prove that the numerical
approximation is well-balanced. We provide numerical examples that verify the
theoretical findings and furthermore provide an application of the scheme for a
partial break of a curved dam test problem
A Provably Stable Discontinuous Galerkin Spectral Element Approximation for Moving Hexahedral Meshes
We design a novel provably stable discontinuous Galerkin spectral element
(DGSEM) approximation to solve systems of conservation laws on moving domains.
To incorporate the motion of the domain, we use an arbitrary
Lagrangian-Eulerian formulation to map the governing equations to a fixed
reference domain. The approximation is made stable by a discretization of a
skew-symmetric formulation of the problem. We prove that the discrete
approximation is stable, conservative and, for constant coefficient problems,
maintains the free-stream preservation property. We also provide details on how
to add the new skew-symmetric ALE approximation to an existing discontinuous
Galerkin spectral element code. Lastly, we provide numerical support of the
theoretical results
Current status of the small punch test standardization within the ASTM
Since 2014, considerable attention has been paid to the standardization of small punch test technique within the American Society of Testing and Materials (ASTM). In 2016 large inter-laboratory study has been launched within the ASTM subcommittee E10.2 - Behavior and Use of Nuclear Structural Materials, involving 12 laboratories and 6 evaluated structural materials from the nuclear power plant components. Paper describes the current status of ASTM standardization, results of the inter-laboratory study, lessons learned and open questions remaining to be solved for the successful completion of the standardization process
H3 histamine receptor-mediated activation of protein kinase calpha inhibits the growth of cholangiocarcinoma in vitro and in vivo
Histamine regulates functions via four receptors (HRH1, HRH2, HRH3, and HRH4). The D-myo-inositol 1,4,5-trisphosphate (IP(3))/Ca(2+)/protein kinase C (PKC)/mitogen-activated protein kinase pathway regulates cholangiocarcinoma growth. We evaluated the role of HRH3 in the regulation of cholangiocarcinoma growth. Expression of HRH3 in intrahepatic and extrahepatic cell lines, normal cholangiocytes, and human tissue arrays was measured. In Mz-ChA-1 cells stimulated with (R)-(alpha)-(-)-methylhistamine dihydrobromide (RAMH), we measured (a) cell growth, (b) IP(3) and cyclic AMP levels, and (c) phosphorylation of PKC and mitogen-activated protein kinase isoforms. Localization of PKC alpha was visualized by immunofluorescence in cell smears and immunoblotting for PKC alpha in cytosol and membrane fractions. Following knockdown of PKC alpha, Mz-ChA-1 cells were stimulated with RAMH before evaluating cell growth and extracellular signal-regulated kinase (ERK)-1/2 phosphorylation. In vivo experiments were done in BALB/c nude mice. Mice were treated with saline or RAMH for 44 days and tumor volume was measured. Tumors were excised and evaluated for proliferation, apoptosis, and expression of PKC alpha, vascular endothelial growth factor (VEGF)-A, VEGF-C, VEGF receptor 2, and VEGF receptor 3. HRH3 expression was found in all cells. RAMH inhibited the growth of cholangiocarcinoma cells. RAMH increased IP(3) levels and PKC alpha phosphorylation and decreased ERK1/2 phosphorylation. RAMH induced a shift in the localization of PKC alpha expression from the cytosolic domain into the membrane region of Mz-ChA-1 cells. Silencing of PKC alpha prevented RAMH inhibition of Mz-ChA-1 cell growth and ablated RAMH effects on ERK1/2 phosphorylation. In vivo, RAMH decreased tumor growth and expression of VEGF and its receptors; PKC alpha expression was increased. RAMH inhibits cholangiocarcinoma growth by PKC alpha-dependent ERK1/2 dephosphorylation. Modulation of PKC alpha by histamine receptors may be important in regulating cholangiocarcinoma growth. (Mol Cancer Res 2009;7(10):1704-13
Insights into the regulation of DMSP synthesis in the diatom Thalassiosira pseudonana through APR activity, proteomics and gene expression analyses on cells acclimating to changes in salinity, light and nitrogen
Despite the importance of dimethylsulphoniopropionate (DMSP) in the global sulphur cycle and climate regulation, the biological pathways underpinning its synthesis in marine phytoplankton remain poorly understood. The intracellular concentration of DMSP increases with increased salinity, increased light intensity and nitrogen starvation in the diatom Thalassiosira pseudonana. We used these conditions to investigate DMSP synthesis at the cellular level via analysis of enzyme activity, gene expression and proteome comparison. The activity of the key sulphur assimilatory enzyme, adenosine 5′- phosphosulphate reductase was not coordinated with increasing intracellular DMSP concentration. Under all three treatments coordination in the expression of sulphur assimilation genes was limited to increases in sulphite reductase transcripts. Similarly, proteomic 2D gel analysis only revealed an increase in phosphoenolpyruvate carboxylase following increases in DMSP concentration. Our findings suggest that increased sulphur assimilation might not be required for increased DMSP synthesis, instead the availability of carbon and nitrogen substrates may be important in the regulation of this pathway. This contrasts with the regulation of sulphur metabolism in higher plants, which generally involves upregulation of several sulphur assimilatory enzymes. In T. pseudonana changes relating to sulphur metabolism were specific to the individual treatments and, given that little coordination was seen in transcript and protein responses across the three growth conditions, different patterns of regulation might be responsible for the increase in DMSP concentration seen under each treatment
Sulphur flux through the sulphate assimilation pathway is differently controlled by adenosine 5′-phosphosulphate reductase under stress and in transgenic poplar plants overexpressing γ-ECS, SO, or APR
Sulphate assimilation provides reduced sulphur for the synthesis of cysteine, methionine, and numerous other essential metabolites and secondary compounds. The key step in the pathway is the reduction of activated sulphate, adenosine 5′-phosphosulphate (APS), to sulphite catalysed by APS reductase (APR). In the present study, [35S]sulphur flux from external sulphate into glutathione (GSH) and proteins was analysed to check whether APR controls the flux through the sulphate assimilation pathway in poplar roots under some stress conditions and in transgenic poplars. (i) O-Acetylserine (OAS) induced APR activity and the sulphur flux into GSH. (ii) The herbicide Acetochlor induced APR activity and results in a decline of GSH. Thereby the sulphur flux into GSH or protein remained unaffected. (iii) Cd treatment increased APR activity without any changes in sulphur flux but lowered sulphate uptake. Several transgenic poplar plants that were manipulated in sulphur metabolism were also analysed. (i) Transgenic poplar plants that overexpressed the γ-glutamylcysteine synthetase (γ-ECS) gene, the enzyme catalysing the key step in GSH formation, showed an increase in sulphur flux into GSH and sulphate uptake when γ-ECS was targeted to the cytosol, while no changes in sulphur flux were observed when γ-ECS was targeted to plastids. (ii) No effect on sulphur flux was observed when the sulphite oxidase (SO) gene from Arabidopsis thaliana, which catalyses the back reaction of APR, that is the reaction from sulphite to sulphate, was overexpressed. (iii) When Lemna minor APR was overexpressed in poplar, APR activity increased as expected, but no changes in sulphur flux were observed. For all of these experiments the flux control coefficient for APR was calculated. APR as a controlling step in sulphate assimilation seems obvious under OAS treatment, in γ-ECS and SO overexpressing poplars. A possible loss of control under certain conditions, that is Cd treatment, Acetochlor treatment, and in APR overexpressing poplar, is discussed
Genetic regulation of glucoraphanin accumulation in Beneforté® broccoli
Diets rich in broccoli (Brassica oleracea var italica) have been associated with maintenance of cardiovascular health and reduction in risk of cancer. These health benefits have been
attributed to glucoraphanin that specifically accumulates in broccoli. The development of broccoli with enhanced concentrations of glucoraphanin may deliver greater health benefits.
Three high-glucoraphanin F1 broccoli hybrids were developed in independent programmes through genome introgression from the wild species Brassica villosa. Glucoraphanin and other metabolites were quantified in experimental field trials. Global SNP analyses quantified the differential extent of B. villosa introgression
The high-glucoraphanin broccoli hybrids contained 2.5–3 times the glucoraphanin content of standard hybrids due to enhanced sulphate assimilation and modifications in sulphur partitioning between sulphur-containing metabolites. All of the high-glucoraphanin hybrids possessed an introgressed B. villosa segment which contained a B. villosa Myb28 allele. Myb28
expression was increased in all of the high-glucoraphanin hybrids. Two high-glucoraphanin hybrids have been commercialised as Beneforte broccoli.
The study illustrates the translation of research on glucosinolate genetics from Arabidopsis to broccoli, the use of wild Brassica species to develop cultivars with potential consumer benefits, and the development of cultivars with contrasting concentrations of glucoraphanin for use in blinded human intervention studie
The sulfur pathway and diagnosis of sulfate depletion in grapevine
Sulfur is an essential nutrient to all plant species. Plants assimilate sulfur
in a well-described pathway, which has been taken up by roots. Regulatory mech-
anism has been the subject of many research papers. However, recent studies
highlighted differences between crop plants and the model plant Arabidopsis
thaliana. Our work focuses on the identification of genes involved in the sulfur
metabolism in the Vitis vinifera genome, and their response to sulfur deficiency and
other abiotic stress endured by grapevine in the field, namely water stress. Here, we
describe the identification and brief characterization of the first assimilation
enzymes involved in the sulfur pathway, the enzyme responsible for sulfur activa-
tion, ATP sulfurylase (ATPS), and the two enzymes that reduce sulfate to sulfide,
Adenosine 50-phosphosulate reductase (APR) and Sulfite reductase (SiR). A reduc-
tion was observed in the number of ATPS and APR isoforms identified in
V. vinifera genome when compared to A. thaliana or Glycine max genomes. Two
ATPS isoforms were present in the Vitis genome, of which only ATPS1 transcript
was detected in the tested tissues, and one APR isoform, suggesting an absence of
redundancy in the role of both enzymes. ATPS1, APR and SiR transcript level was
up-regulated in response to 2 days exposure to sulfur deficiency in V. vinifera cell
cultures, which was completely reversed by the addition of GSH to the culture
medium. Apparently, oxidative stress triggered GSH has a pivotal role in the
regulation of ATPS1, APR and SiR transcription level, since their up-regulation
was observed in mRNA from field grapevine berries under water stress, which is
known to induce oxidative stress.info:eu-repo/semantics/publishedVersio
The bundle sheath of rice is conditioned to play an active role in water transport as well as sulfur assimilation and jasmonic acid synthesis
Funder: Bill and Melinda Gates Foundation; Id: http://dx.doi.org/10.13039/100000865Summary: Leaves comprise multiple cell types but our knowledge of the patterns of gene expression that underpin their functional specialization is fragmentary. Our understanding and ability to undertake the rational redesign of these cells is therefore limited. We aimed to identify genes associated with the incompletely understood bundle sheath of C3 plants, which represents a key target associated with engineering traits such as C4 photosynthesis into Oryza sativa (rice). To better understand the veins, bundle sheath and mesophyll cells of rice, we used laser capture microdissection followed by deep sequencing. Gene expression of the mesophyll is conditioned to allow coenzyme metabolism and redox homeostasis, as well as photosynthesis. In contrast, the bundle sheath is specialized in water transport, sulphur assimilation and jasmonic acid biosynthesis. Despite the small chloroplast compartment of bundle sheath cells, substantial photosynthesis gene expression was detected. These patterns of gene expression were not associated with the presence or absence of specific transcription factors in each cell type, but were instead associated with gradients in expression across the leaf. Comparative analysis with C3 Arabidopsis identified a small gene set preferentially expressed in the bundle sheath cells of both species. This gene set included genes encoding transcription factors from 14 orthogroups and proteins allowing water transport, sulphate assimilation and jasmonic acid synthesis. The most parsimonious explanation for our findings is that bundle sheath cells from the last common ancestor of rice and Arabidopsis were specialized in this manner, and as the species diverged these patterns of gene expression have been maintained
Deficiency in the Phosphorylated Pathway of Serine Biosynthesis Perturbs Sulfur Assimilation
Anoman AD, Flores-Tornero M, Benstein RM, et al. Deficiency in the Phosphorylated Pathway of Serine Biosynthesis Perturbs Sulfur Assimilation. PLANT PHYSIOLOGY. 2019;180(1):153-170.Although the plant Phosphorylated Pathway of L-Ser Biosynthesis (PPSB) is essential for embryo and pollen development, and for root growth, its metabolic implications have not been fully investigated. A transcriptomics analysis of Arabidopsis (Arabidopsis thaliana) PPSB-deficient mutants at night, when PPSB activity is thought to be more important, suggested interaction with the sulfate assimilation process. Because sulfate assimilation occurs mainly in the light, we also investigated it in PPSB-deficient lines in the day. Key genes in the sulfate starvation response, such as the adenosine 5'phosphosulfate reductase genes, along with sulfate transporters, especially those involved in sulfate translocation in the plant, were induced in the PPSB-deficient lines. However, sulfate content was not reduced in these lines as compared with wild-type plants; besides the glutathione (GSH) steady-state levels in roots of PPSB-deficient lines were even higher than in wild type. This suggested that PPSB deficiency perturbs the sulfate assimilation process between tissues/organs. Alteration of thiol distribution in leaves from different developmental stages, and between aerial parts and roots in plants with reduced PPSB activity, provided evidence supporting this idea. Diminished PPSB activity caused an enhanced flux of S-35 into thiol biosynthesis, especially in roots. GSH turnover also accelerated in the PPSB-deficient lines, supporting the notion that not only biosynthesis, but also transport and allocation, of thiols were perturbed in the PPSB mutants. Our results suggest that PPSB is required for sulfide assimilation in specific heterotrophic tissues and that a lack of PPSB activity perturbs sulfur homeostasis between photosynthetic and nonphotosynthetic tissues
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