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Sample records for sugar-1-phosphate nucleotidyltransferase assay

  1. Mechanism of Mg2+-Accompanied Product Release in Sugar Nucleotidyltransferases.

    Science.gov (United States)

    Vithani, Neha; Ankush Jagtap, Pravin Kumar; Verma, Sunil Kumar; Tripathi, Ravi; Awasthi, Shalini; Nair, Nisanth N; Prakash, Balaji

    2018-03-06

    The nucleotidyl transfer reaction, catalyzed by sugar nucleotidyltransferases (SNTs), is assisted by two active site Mg 2+ ions. While studying this reaction using X-ray crystallography, we captured snapshots of the pyrophosphate (product) as it exits along a pocket. Surprisingly, one of the active site Mg 2+ ions remains coordinated to the exiting pyrophosphate. This hints at the participation of Mg 2+ in the process of product release, besides its role in catalyzing nucleotidyl transfer. These observations are further supported by enhanced sampling molecular dynamics simulations. Free energy computations suggest that the product release is likely to be rate limiting in SNTs, and the origin of the high free energy barrier for product release could be traced back to the "slow" conformational change of an Arg residue at the exit end of the pocket. These results establish a dual role for Mg 2+ , and propose a general mechanism of product release during the nucleotidyl transfer by SNTs. Copyright © 2018 Elsevier Ltd. All rights reserved.

  2. Sphingosine 1-phosphate lyase enzyme assay using a BODIPY-labeled substrate

    International Nuclear Information System (INIS)

    Bandhuvula, Padmavathi; Li Zaiguo; Bittman, Robert; Saba, Julie D.

    2009-01-01

    Sphingosine 1-phosphate lyase (SPL) is responsible for the irreversible catabolism of sphingosine 1-phosphate, which signals through five membrane receptors to mediate cell stress responses, angiogenesis, and lymphocyte trafficking. The standard assay for SPL activity utilizes a radioactive dihydrosphingosine 1-phosphate substrate and is expensive and cumbersome. In this study, we describe an SPL assay that employs an ω-labeled BODIPY-sphingosine 1-phosphate substrate, allowing fluorescent product detection by HPLC and incorporating advantages of the BODIPY fluorophore. The major aldehyde product is confirmed by reaction with 2,4-dinitrophenylhydrazine. The SPL-catalyzed reaction is linear over a 30 min time period and yields a K m of 35 μM for BODIPY-sphingosine 1-phosphate.

  3. Vanadate influence on metabolism of sugar phosphates in fungus Phycomyces blakesleeanus.

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    Milan Žižić

    Full Text Available The biological and chemical basis of vanadium action in fungi is relatively poorly understood. In the present study, we investigate the influence of vanadate (V5+ on phosphate metabolism of Phycomyces blakesleeanus. Addition of V5+ caused increase of sugar phosphates signal intensities in 31P NMR spectra in vivo. HPLC analysis of mycelial phosphate extracts demonstrated increased concentrations of glucose 6 phosphate, fructose 6 phosphate, fructose 1, 6 phosphate and glucose 1 phosphate after V5+ treatment. Influence of V5+ on the levels of fructose 2, 6 phosphate, glucosamine 6 phosphate and glucose 1, 6 phosphate (HPLC, and polyphosphates, UDPG and ATP (31P NMR was also established. Increase of sugar phosphates content was not observed after addition of vanadyl (V4+, indicating that only vanadate influences its metabolism. Obtained results from in vivo experiments indicate catalytic/inhibitory vanadate action on enzymes involved in reactions of glycolysis and glycogenesis i.e., phosphoglucomutase, phosphofructokinase and glycogen phosphorylase in filamentous fungi.

  4. Structure-guided Mutational Analysis of the Nucleotidyltransferase Domain of Escherichia coli DNA Ligase (LigA).

    Science.gov (United States)

    Wang, Li Kai; Zhu, Hui; Shuman, Stewart

    2009-03-27

    NAD(+)-dependent DNA ligases (LigA) are ubiquitous in bacteria, where they are essential for growth and present attractive targets for antimicrobial drug discovery. LigA has a distinctive modular structure in which a nucleotidyltransferase catalytic domain is flanked by an upstream NMN-binding module and by downstream OB-fold, zinc finger, helix-hairpin-helix, and BRCT domains. Here we conducted a structure-function analysis of the nucleotidyltransferase domain of Escherichia coli LigA, guided by the crystal structure of the LigA-DNA-adenylate intermediate. We tested the effects of 29 alanine and conservative mutations at 15 amino acids on ligase activity in vitro and in vivo. We thereby identified essential functional groups that coordinate the reactive phosphates (Arg(136)), contact the AMP adenine (Lys(290)), engage the phosphodiester backbone flanking the nick (Arg(218), Arg(308), Arg(97) plus Arg(101)), or stabilize the active domain fold (Arg(171)). Finer analysis of the mutational effects revealed step-specific functions for Arg(136), which is essential for the reaction of LigA with NAD(+) to form the covalent ligase-AMP intermediate (step 1) and for the transfer of AMP to the nick 5'-PO(4) to form the DNA-adenylate intermediate (step 2) but is dispensable for phosphodiester formation at a preadenylylated nick (step 3).

  5. Identification of residues involved in nucleotidyltransferase activity of JHP933 from helicobacter pyloriby site-directed mutagenesis

    Directory of Open Access Journals (Sweden)

    Ye Xianren

    2016-01-01

    Full Text Available Helicobacter pylori is a well-known bacterial pathogen involved in the development of peptic ulcer, gastric adenocarcinoma and other forms of gastric cancer. Evidence has suggested that certain strain-specific genes in the plasticity region may play key roles in the pathogenesis of H. pylori-associated gastroduodenal diseases. Therefore there is considerable interest in the strain-specific genes located in the plasticity regions of H. pylori. JHP933 is encoded by the gene in the plasticity region of H. pylori strain J99. Recently, the crystal structure of JHP933 has confirmed it as a nucleotidyltransferase (NTase superfamily protein and a putative active site has been proposed. However, no evidence from direct functional assay has been presented to confirm the active site and little is known about the functional mechanism of JHP933. Here, through superimposition with Cid1/NTP complex structures, we modelled the complex structures of JHP933 with different NTPs. Based on the models and using rational site-directed mutagenesis combined with enzymatic activity assays, we confirm the active site and identify several residues important for the nucleotidyl transferring function of JHP933. Furthermore, mutations of these active site residues result in the abolishment of the nucleotidyltransferase activity of JHP933. This work provides preliminary insight into the molecular mechanism underlying the pathophysiological role in H. pylori infection of JHP933 as a novel NTase superfamily protein.

  6. Coordination chemistry of sugar-phosphate complexes with palladium(II), rhenium(V) and zinc(II)

    Energy Technology Data Exchange (ETDEWEB)

    Steinborn, Christian Martin

    2013-05-21

    As described before, some studies dealing with coordination chemistry of sugar phosphates are available but no analogous complexes of Zn{sup II} have been investigated yet. The primary goal of this work is, therefore, to fill this gap. In order to stay close to the active sites of enzymes such as class-II-aldolase, the simple metal fragment Zn{sup II}(dien) is used. NMR spectroscopy is used primarily as analytical method since it enables the investigation of both complex equilibria in solution and pH dependence of metal-binding sites. Since this approach is challenging due to the fast metal-ligand exchange and the absence of CIS values, it is necessary to improve the significance of NMR data collected from sugar-phosphate complexes with Zn{sup II}. Hence, further experiments are performed with molecules similar to sugar phosphates such as reducing and methylated sugars or polyols. Beside NMR spectroscopy, crystal-structure analysis will be used to get more detailed information about the binding pattern of the complexes. Additionally, sugar-phosphate complexes of Pd{sup II} are investigated. Further experiments are conducted, on the one hand, to synthesise more sugarphosphate complexes with ReVON2 fragments, and, on the other hand, to grow crystals confirming the theory about mixed sugar-core-phosphate chelation.

  7. Enzymatic synthesis of rare sugars with L-rhamnulose-1-phosphate aldolase from Thermotoga maritima MSB8.

    Science.gov (United States)

    Li, Zijie; Wu, Xiaoru; Cai, Li; Duan, Shenglin; Liu, Jia; Yuan, Peng; Nakanishi, Hideki; Gao, Xiao-Dong

    2015-09-15

    L-Rhamnulose-1-phosphate aldolase from a thermophilic source (Thermotoga maritima MSB8) (RhaDT.mari) was heterologously overexpressed in Escherichia coli and the stereoselectivity of this enzyme with or without Nus tag was investigated. We also applied this enzyme to the synthesis of rare sugars D-psicose, D-sorbose, L-tagatose and L-fructose using our one-pot four-enzyme system. To the best of our knowledge, this is the first use of RhaD from a thermophilic source for rare sugar synthesis and the temperature tolerance of this enzyme paves the path for large scale fermentation. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Structure-based prediction and identification of 4-epimerization activity of phosphate sugars in class II aldolases.

    Science.gov (United States)

    Lee, Seon-Hwa; Hong, Seung-Hye; An, Jung-Ung; Kim, Kyoung-Rok; Kim, Dong-Eun; Kang, Lin-Woo; Oh, Deok-Kun

    2017-05-16

    Sugar 4-epimerization reactions are important for the production of rare sugars and their derivatives, which have various potential industrial applications. For example, the production of tagatose, a functional sweetener, from fructose by sugar 4-epimerization is currently constrained because a fructose 4-epimerase does not exist in nature. We found that class II D-fructose-1,6-bisphosphate aldolase (FbaA) catalyzed the 4-epimerization of D-fructose-6-phosphate (F6P) to D-tagatose-6-phosphate (T6P) based on the prediction via structural comparisons with epimerase and molecular docking and the identification of the condensed products of C3 sugars. In vivo, the 4-epimerization activity of FbaA is normally repressed. This can be explained by our results showing the catalytic efficiency of D-fructose-6-phosphate kinase for F6P phosphorylation was significantly higher than that of FbaA for F6P epimerization. Here, we identified the epimerization reactions and the responsible catalytic residues through observation of the reactions of FbaA and L-rhamnulose-1-phosphate aldolases (RhaD) variants with substituted catalytic residues using different substrates. Moreover, we obtained detailed potential epimerization reaction mechanism of FbaA and a general epimerization mechanism of the class II aldolases L-fuculose-1-phosphate aldolase, RhaD, and FbaA. Thus, class II aldolases can be used as 4-epimerases for the stereo-selective synthesis of valuable carbohydrates.

  9. Sensitive, specific radioisotope assay for L-glutamine-D-fructose-6- phosphat aminotransferase

    International Nuclear Information System (INIS)

    Callahan, M.; Tourian, A.; Hung, W.Y.

    1981-01-01

    A sensitive and specific radioassay for L-glutamine-D-fructose-6-phosphate aminotransferase (EC 5.3.1.19) activity is presented. Picomoles of product are measurable, and the assay can be applied to systems having limited quantities of available protein, particularly in extracts of either cell or organ cultures. The assay is at least 10,000 times more sensitive under K 1 concentrations of fructose 6-phosphate than the modified Elson-Morgan colorimetric assay and 20 times more sensitive under saturating conditions of fructose 6-phosphate. As little as 0.5 μg of cell-extract protein will yield measurable product. In contrast, 280 μg of crude-extract protein from colon is required with the modified Elson-Morgan colorimetric assay

  10. Increasing the Thermostable Sugar-1-Phosphate Nucleotidylyltransferase Activities of the Archaeal ST0452 Protein through Site Saturation Mutagenesis of the 97th Amino Acid Position.

    Science.gov (United States)

    Honda, Yuki; Zang, Qian; Shimizu, Yasuhiro; Dadashipour, Mohammad; Zhang, Zilian; Kawarabayasi, Yutaka

    2017-02-01

    The ST0452 protein is a bifunctional protein exhibiting sugar-1-phosphate nucleotidylyltransferase (sugar-1-P NTase) and amino-sugar-1-phosphate acetyltransferase activities and was isolated from the thermophilic archaeon Sulfolobus tokodaii Based on the previous observation that five single mutations increased ST0452 sugar-1-P NTase activity, nine double-mutant ST0452 proteins were generated with the intent of obtaining enzymes exhibiting a further increase in catalysis, but all showed less than 15% of the wild-type N-acetyl-d-glucosamine-1-phosphate uridyltransferase (GlcNAc-1-P UTase) activity. The Y97A mutant exhibited the highest activity of the single-mutant proteins, and thus site saturation mutagenesis of the 97th position (Tyr) was conducted. Six mutants showed both increased GlcNAc-1-P UTase and glucose-1-phosphate uridyltransferase activities, eight mutants showed only enhanced GlcNAc-1-P UTase activity, and six exhibited higher GlcNAc-1-P UTase activity than that of the Y97A mutant. Kinetic analyses of three typical mutants indicated that the increase in sugar-1-P NTase activity was mainly due to an increase in the apparent k cat value. We hypothesized that changing the 97th position (Tyr) to a smaller amino acid with similar electronic properties would increase activity, and thus the Tyr at the corresponding 103rd position of the Escherichia coli GlmU (EcGlmU) enzyme was replaced with the same residues. The Y103N mutant EcGlmU showed increased GlcNAc-1-P UTase activity, revealing that the Tyr at the 97th position of the ST0452 protein (103rd position in EcGlmU) plays an important role in catalysis. The present results provide useful information regarding how to improve the activity of natural enzymes and how to generate powerful enzymes for the industrial production of sugar nucleotides. It is typically difficult to increase enzymatic activity by introducing substitutions into a natural enzyme. However, it was previously found that the ST0452 protein

  11. Use of osmolytes during solubilization and reconstitution of phosphate: sugar phosphate antiport from bacteria

    International Nuclear Information System (INIS)

    Ambudkar, S.V.; Sonna, L.A.; Maloney, P.C.

    1986-01-01

    Phosphate:2-deoxyglucose 6-phosphate (Pi:2DG6P) antiport was extracted from Streptococcus lactis or Staphylococcus aureus with 1.1% octylglucoside in the presence of 0.37% E. coli lipid and reconstituted by detergent dilution. Because previous work suggested inactivation at an early stage, the authors introduced protein stabilants during solubilization. When 20% glycerol was used, proteoliposomes showed a 20-fold increase in 32 Pi transport. This enhanced recovery required phospholipid plus glycerol, and was found only when both were added together with the detergent. Glycerol protection yielded proteoliposomes in which antiporters retained their normal kinetic properties, and Pi exchange by the streptococcal example gave a maximal rate (200-400 nmol/min per mg protein) and a turnover number (30-50/s) which suggested that inactivation had been avoided. Further study showed that 20% glycerol could be replaced by equally high concentrations of compounds classified as osmolytes polyols (erythritol, xylitol, sorbitol), sugars (glucose, trehalose) and certain amino acids (glycine, proline, but not valine). The authors suggest that osmolytes may be used to fully stabilize chemiosmotic transporters during reconstitution

  12. Analysis of phosphate esters in plant material. Extraction and purification.

    Science.gov (United States)

    Isherwood, F A; Barrett, F C

    1967-09-01

    1. A critical study was made of the quantitative extraction of nucleotide and sugar phosphates from plant tissue by either boiling aqueous ethanol or cold trichloroacetic acid. The effect of the extraction technique on the inactivation of the enzymes in the plant tissue and the possibility of adsorption of the phosphate esters on the cell wall were especially considered. 2. In the recommended method the plant tissue was frozen in liquid nitrogen, ground to a powder and then blended with cold aqueous trichloroacetic acid containing 8-hydroxyquinoline to prevent adsorption. 3. The extract contained large amounts of trichloroacetic acid, cations, chloride, sugars, amino acids, hydroxy organic acids, phytic acid, orthophosphoric acid and high-molecular-weight material including some phosphorus-containing compounds. All of these were removed as they were liable to interfere with the chromatographic or enzymic assay of the individual nucleotide or sugar phosphates. 4. The procedure was as follows: the last traces of trichloroacetic acid were extracted with ether after the solution had been passed through a column of Dowex AG 50 in the hydrogen form to remove all cations. High-molecular-weight compounds were removed by ultrafiltration and low-molecular-weight solutes by a two-stage chromatography on cellulose columns with organic solvents. In the first stage, sugars, amino acids, chloride and phytic acid were separated by using a basic solvent (propan-1-ol-water-aqueous ammonia) and, in the second stage, the organic acids and orthophosphoric acid were separated by using an acidic solvent (di-isopropyl ether-formic acid-2-methylpropan-2-ol-water). The final solution of nucleotide and sugar phosphates was substantially free from other solutes and was suitable for the detection of individual phosphate esters by either chromatography or enzymic assay. 5. The recovery of d-glucose 6-phosphate or adenosine 5'-triphosphate added to a trichloroacetic acid extract simulating that

  13. Structure-guided mutational analysis of the nucleotidyltransferase domain of Escherichia coli NAD+-dependent DNA ligase (LigA).

    Science.gov (United States)

    Zhu, Hui; Shuman, Stewart

    2005-04-01

    NAD+-dependent DNA ligase (LigA) is essential for bacterial growth and a potential target for antimicrobial drug discovery. Here we queried the role of 14 conserved amino acids of Escherichia coli LigA by alanine scanning and thereby identified five new residues within the nucleotidyltransferase domain as being essential for LigA function in vitro and in vivo. Structure activity relationships were determined by conservative mutagenesis for the Glu-173, Arg-200, Arg-208, and Arg-277 side chains, as well as four other essential side chains that had been identified previously (Lys-115, Asp-117, Asp-285, and Lys-314). In addition, we identified Lys-290 as important for LigA activity. Reference to the structure of Enterococcus faecalis LigA allowed us to discriminate three classes of essential/important side chains that: (i) contact NAD+ directly (Lys-115, Glu-173, Lys-290, and Lys-314); (ii) comprise the interface between the NMN-binding domain (domain Ia) and the nucleotidyltransferase domain or comprise part of a nick-binding site on the surface of the nucleotidyltransferase domain (Arg-200 and Arg-208); or (iii) stabilize the active site fold of the nucleotidyltransferase domain (Arg-277). Analysis of mutational effects on the isolated ligase adenylylation and phosphodiester formation reactions revealed different functions for essential side chains at different steps of the DNA ligase pathway, consistent with the proposal that the active site is serially remodeled as the reaction proceeds.

  14. Sucrose Phosphate Synthase and Sucrose Accumulation at Low Temperature 1

    Science.gov (United States)

    Guy, Charles L.; Huber, Joan L. A.; Huber, Steven C.

    1992-01-01

    The influence of growth temperature on the free sugar and sucrose phosphate synthase content and activity of spinach (Spinacia oleracea) leaf tissue was studied. When plants were grown at 25°C for 3 weeks and then transferred to a constant 5°C, sucrose, glucose, and fructose accumulated to high levels during a 14-d period. Predawn sugar levels increased from 14- to 20-fold over the levels present at the outset of the low-temperature treatment. Sucrose was the most abundant free sugar before, during, and after exposure to 5°C. Leaf sucrose phosphate synthase activity was significantly increased by the low-temperature treatment, whereas sucrose synthase and invertases were not. Synthesis of the sucrose phosphate synthase subunit was increased during and after low-temperature exposure and paralleled an increase in the steady-state level of the subunit. The increases in sucrose and its primary biosynthetic enzyme, sucrose phosphate synthase, are discussed in relation to adjustment of metabolism to low nonfreezing temperature and freezing stress tolerance. Images Figure 1 Figure 2 Figure 3 PMID:16652990

  15. Synthesis of rare sugars with L-fuculose-1-phosphate aldolase (FucA) from Thermus thermophilus HB8.

    Science.gov (United States)

    Li, Zijie; Cai, Li; Qi, Qingsheng; Styslinger, Thomas J; Zhao, Guohui; Wang, Peng George

    2011-09-01

    We report herein a one-pot four-enzyme approach for the synthesis of the rare sugars d-psicose, d-sorbose, l-tagatose, and l-fructose with aldolase FucA from a thermophilic source (Thermus thermophilus HB8). Importantly, the cheap starting material DL-GP (DL-glycerol 3-phosphate), was used to significantly reduce the synthetic cost. Copyright © 2011 Elsevier Ltd. All rights reserved.

  16. Dynamics of sugar-metabolic enzymes and sugars accumulation during watermelon (citrullus lanatus) fruit development

    International Nuclear Information System (INIS)

    Zhang, H.

    2016-01-01

    We analyzed sugar accumulation and the activities of sugar-metabolic enzymes in ripening fruits of three cultivars of watermelon; a high-sugar type w2, a low-sugar type (w1), and their hybrid. In w2, the glucose and fructose contents were higher than the sucrose content in the earlier stage of fruit development, and fruit growth was accompanied by increases in glucose, fructose, and sucrose contents. The sucrose content increased substantially after 20 days after anthesis (DAA) and it was the main soluble sugar in mature fruit (sucrose: hexoses ratio, 0.71). In W, the fructose and glucose contents were significantly higher than the sucrose content in mature fruit (sucrose: hexoses ratio, 0.25). Comparing the two parent cultivars, sucrose was the most important factor affecting the total sugar content in mature fruit, although glucose and fructose also contributed to total sugar contents. The fructose and glucose contents in the fruit of F1 were mid-way between those of their parents, while the sucrose content was closer to that of W (sucrose:hexoses ratio in F1, 0.26). In the early stage of fruit development of W2, the activities of acid invertase and neutral invertase were higher than those of sucrose synthase and sucrose phosphate synthase. After 20 DAA, the acid invertase and neutral invertase activities decreased and those of sucrose synthase and sucrose phosphate synthase increased, leading to increased sucrose content. In W1, the activities of acid invertase and neutral invertase were higher than those of sucrose synthase and sucrose phosphate synthase at the early stage. The sucrose synthase and sucrose phosphate synthase activities were lower in W1 than in W2 at the later stages of fruit development. The patterns of sugar accumulation and sugar-metabolic enzyme activities during fruit development in F1 were similar to those in W1. (author)

  17. Sugar signaling regulation by Arabidopsis SIZ1-driven sumoylation is independent of salicylic acid

    DEFF Research Database (Denmark)

    Castro, Pedro Humberto Araújo R F; Verde, Nuno; Tavares, Rui Manuel

    2018-01-01

    inefficient responses to nutrient imbalance in phosphate, nitrate and copper. Recently, we reported that siz1 also displays altered responses to exogenous sugar supplementation. The siz1 mutant is a salicylic acid (SA) accumulator, and SA may interfere with sugar-dependent responses and signaling events. Here......, we extended our previous studies to determine the importance of SA in the SIZ1 response to sugars, by introducing the bacterial salicylate hydroxylase NahG into the siz1 background. Results demonstrate that siz1 phenotypes involving delayed germination are partially dependent of SA levels, whereas...

  18. Dinuclear copper(II) complexes with {Cu2(mu-hydroxo)bis(mu-carboxylato)}+ cores and their reactions with sugar phosphate esters: A substrate binding model of fructose-1,6-bisphosphatase.

    Science.gov (United States)

    Kato, Merii; Tanase, Tomoaki; Mikuriya, Masahiro

    2006-04-03

    Reactions of CuX2.nH2O with the biscarboxylate ligand XDK (H2XDK = m-xylenediamine bis(Kemp's triacid imide)) in the presence of N-donor auxiliary ligands yielded a series of dicopper(II) complexes, [Cu2(mu-OH)(XDK)(L)2]X (L = N,N,N',N'-tetramethylethylenediamine (tetmen), X = NO3 (1a), Cl (1b); L = N,N,N'-trimethylethylenediamine (tmen), X = NO3 (2a), Cl (2b); L =2,2'-bipyridine (bpy), X = NO3 (3); L = 1,10-phenanthroline (phen), X = NO3 (4); L = 4,4'-dimethyl-2,2'-bipyridine (Me2bpy), X = NO3 (5); L = 4-methyl-1,10-phenanthroline (Mephen), X = NO3 (6)). Complexes 1-6 were characterized by X-ray crystallography (Cu...Cu = 3.1624(6)-3.2910(4) A), and the electrochemical and magnetic properties were also examined. Complexes 3 and 4 readily reacted with diphenyl phosphoric acid (HDPP) or bis(4-nitrophenyl) phosphoric acid (HBNPP) to give [Cu2(mu-phosphate)(XDK)(L)2]NO3 (L = bpy, phosphate = DPP (11); L = phen, phosphate = DPP (12), BNPP (13)), where the phsophate diester bridges the two copper ions in a mu-1,3-O,O' bidentate fashion (Cu...Cu = 4.268(3)-4.315(1) A). Complexes 4 and 6 with phen and Mephen have proven to be good precursors to accommodate a series of sugar monophosphate esters (Sugar-P) onto the biscarboxylate-bridged dicopper centers, yielding [Cu2(mu-Sugar-P)(XDK)(L)2] (Sugar-P = alpha-D-Glc-1-P (23a and b), D-Glc-6-P (24a and b), D-Man-6-P (25a), D-Fru-6-P (26a and b); L = phen (a), Mephen (b)) and [Cu2(mu-Gly-n-P)(XDK)(Mephen)2] (Gly-n-P = glycerol n-phosphate; n = 2 (21), 3 (22)), where Glc, Man, and Fru are glucose, mannose, and fructose, respectively. The structure of [Cu2(mu-MNPP)(XDK)(phen)2(CH3OH)] (20) was characterized as a reference compound (H2MNPP = 4-nitrophenyl phosphoric acid). Complexes 4 and 6 also reacted with d-fructose 1,6-bisphosphate (D-Fru-1,6-P2) to afford the tetranuclear copper(II) complexes formulated as [Cu4(mu-D-Fru-1,6-P2)(XDK)2(L)4] (L = phen (27a), Mephen (27b)). The detailed structure of 27a was determined by X

  19. An immunochemical assay for 8,5'-cyclonucleotides in irradiated nucleic acids

    International Nuclear Information System (INIS)

    Fuciarelli, A.F.; Raleigh, J.A.

    1985-01-01

    The transfer of radiation damage initiated in the sugar phosphate backbone to a nucleotide base as exemplified by 8,5'-cyclonucleotide formation has been investigated in polyadenylic acid, native and heat-denatured DNA. Polyclonal antiserum was raised in rabbits with a protein-8,5'-cycloadenosine-5'monophosphate (8,5'-cycloAMP) conjugate prepared by the carbodiimide method. An indirect, enzyme-linked immunosorbent assay (ELISA) was developed with this antiserum to probe for 8,5'-cycloAMP formation. The assay can readily detect product formation in polyadenylic acid irradiated to a total dose of 1.0 krad in the absence of oxygen. Product formation in native or heat-denatured DNA irradiated in 0.1 M phosphate buffer (pH 7.00) in the absence of oxygen is detected after approximately 20 krads. The authors shall extend these studies to determine the utility of immunochemical assays for investigating the radiation chemistry of nucleic acids

  20. Functional Dissection of the DNA Interface of the Nucleotidyltransferase Domain of Chlorella Virus DNA Ligase*

    Science.gov (United States)

    Samai, Poulami; Shuman, Stewart

    2011-01-01

    Chlorella virus DNA ligase (ChVLig) has pluripotent biological activity and an intrinsic nick-sensing function. ChVLig consists of three structural modules that envelop nicked DNA as a C-shaped protein clamp: a nucleotidyltransferase (NTase) domain and an OB domain (these two are common to all DNA ligases) as well as a distinctive β-hairpin latch module. The NTase domain, which performs the chemical steps of ligation, binds the major groove flanking the nick and the minor groove on the 3′-OH side of the nick. Here we performed a structure-guided mutational analysis of the NTase domain, surveying the effects of 35 mutations in 19 residues on ChVLig activity in vivo and in vitro, including biochemical tests of the composite nick sealing reaction and of the three component steps of the ligation pathway (ligase adenylylation, DNA adenylylation, and phosphodiester synthesis). The results highlight (i) key contacts by Thr-84 and Lys-173 to the template DNA strand phosphates at the outer margins of the DNA ligase footprint; (ii) essential contacts of Ser-41, Arg-42, Met-83, and Phe-75 with the 3′-OH strand at the nick; (iii) Arg-176 phosphate contacts at the nick and with ATP during ligase adenylylation; (iv) the role of Phe-44 in forming the protein clamp around the nicked DNA substrate; and (v) the importance of adenine-binding residue Phe-98 in all three steps of ligation. Kinetic analysis of single-turnover nick sealing by ChVLig-AMP underscored the importance of Phe-75-mediated distortion of the nick 3′-OH nucleoside in the catalysis of DNA 5′-adenylylation (step 2) and phosphodiester synthesis (step 3). Induced fit of the nicked DNA into a distorted conformation when bound within the ligase clamp may account for the nick-sensing capacity of ChVLig. PMID:21335605

  1. Functional dissection of the DNA interface of the nucleotidyltransferase domain of chlorella virus DNA ligase.

    Science.gov (United States)

    Samai, Poulami; Shuman, Stewart

    2011-04-15

    Chlorella virus DNA ligase (ChVLig) has pluripotent biological activity and an intrinsic nick-sensing function. ChVLig consists of three structural modules that envelop nicked DNA as a C-shaped protein clamp: a nucleotidyltransferase (NTase) domain and an OB domain (these two are common to all DNA ligases) as well as a distinctive β-hairpin latch module. The NTase domain, which performs the chemical steps of ligation, binds the major groove flanking the nick and the minor groove on the 3'-OH side of the nick. Here we performed a structure-guided mutational analysis of the NTase domain, surveying the effects of 35 mutations in 19 residues on ChVLig activity in vivo and in vitro, including biochemical tests of the composite nick sealing reaction and of the three component steps of the ligation pathway (ligase adenylylation, DNA adenylylation, and phosphodiester synthesis). The results highlight (i) key contacts by Thr-84 and Lys-173 to the template DNA strand phosphates at the outer margins of the DNA ligase footprint; (ii) essential contacts of Ser-41, Arg-42, Met-83, and Phe-75 with the 3'-OH strand at the nick; (iii) Arg-176 phosphate contacts at the nick and with ATP during ligase adenylylation; (iv) the role of Phe-44 in forming the protein clamp around the nicked DNA substrate; and (v) the importance of adenine-binding residue Phe-98 in all three steps of ligation. Kinetic analysis of single-turnover nick sealing by ChVLig-AMP underscored the importance of Phe-75-mediated distortion of the nick 3'-OH nucleoside in the catalysis of DNA 5'-adenylylation (step 2) and phosphodiester synthesis (step 3). Induced fit of the nicked DNA into a distorted conformation when bound within the ligase clamp may account for the nick-sensing capacity of ChVLig.

  2. Molecular data for a biochemical model of DNA damage: Electron impact ionization and dissociative ionization cross sections of DNA bases and sugar-phosphate backbone

    International Nuclear Information System (INIS)

    Huo, Winifred M.; Dateo, Christopher E.; Fletcher, Graham D.

    2006-01-01

    As part of the database for building up a biochemical model of DNA radiation damage, electron impact ionization cross sections of sugar-phosphate backbone and DNA bases have been calculated using the improved binary-encounter dipole (iBED) model. It is found that the total ionization cross sections of C 3 ' - and C 5 ' -deoxyribose-phosphate, two conformers of the sugar-phosphate backbone, are close to each other. Furthermore, the sum of the ionization cross sections of the separate deoxyribose and phosphate fragments is in close agreement with the C 3 ' - and C 5 ' -deoxyribose-phosphate cross sections, differing by less than 10%, an indication that a building-up principle may be applicable. Of the four DNA bases, the ionization cross section of guanine is the largest, then in decreasing order, adenine, thymine, and cytosine. The order is in accordance with the known propensity of oxidation of the bases by ionizing radiation. Dissociative ionization (DI), a process that both ionizes and dissociates a molecule, is investigated for cytosine. The DI cross section for the formation of H and (cytosine-H1) + , with the cytosine ion losing H at the 1 position, is also reported. The threshold of this process is calculated to be 16.9eV. Detailed analysis of ionization products such as in DI is important to trace the sequential steps in the biochemical process of DNA damage

  3. Structural and biochemical characterization of the cell fate determining nucleotidyltransferase fold protein MAB21L1.

    Science.gov (United States)

    de Oliveira Mann, Carina C; Kiefersauer, Reiner; Witte, Gregor; Hopfner, Karl-Peter

    2016-06-08

    The exceptionally conserved metazoan MAB21 proteins are implicated in cell fate decisions and share considerable sequence homology with the cyclic GMP-AMP synthase. cGAS is the major innate immune sensor for cytosolic DNA and produces the second messenger 2'-5', 3'-5' cyclic GMP-AMP. Little is known about the structure and biochemical function of other proteins of the cGAS-MAB21 subfamily, such as MAB21L1, MAB21L2 and MAB21L3. We have determined the crystal structure of human full-length MAB21L1. Our analysis reveals high structural conservation between MAB21L1 and cGAS but also uncovers important differences. Although monomeric in solution, MAB21L1 forms a highly symmetric double-pentameric oligomer in the crystal, raising the possibility that oligomerization could be a feature of MAB21L1. In the crystal, MAB21L1 is in an inactive conformation requiring a conformational change - similar to cGAS - to develop any nucleotidyltransferase activity. Co-crystallization with NTP identified a putative ligand binding site of MAB21 proteins that corresponds to the DNA binding site of cGAS. Finally, we offer a structure-based explanation for the effects of MAB21L2 mutations in patients with eye malformations. The underlying residues participate in fold-stabilizing interaction networks and mutations destabilize the protein. In summary, we provide a first structural framework for MAB21 proteins.

  4. Radical-induced dephosphorylation of fructose phosphates in aqueous solution

    International Nuclear Information System (INIS)

    Zegota, H.; Sonntag, C. von

    1981-01-01

    Oxygen free N 2 O-saturated aqueous solutions of D-fructose-1-phosphate and D-fructose-6-phosphate were γ-irradiated. Inorganic phosphate and phosphate free sugars (containing four to six carbon atoms) were identified and their G-values measured. D-Fructose-1-phosphate yields (G-values in parentheses) inorganic phosphate (1.6), hexos-2-ulose (0.12), 6-deoxy-2,5-hexodiulose (0.16), tetrulose (0.05) and 3-deoxytetrulose (0.15). D-Fructose-6-phosphate yields inorganic phosphate (1.7), hexos-5-ulose (0.1), 6-deoxy-2,5-hexodiulose (0.36), 3-deoxy-2,5-hexodiulose and 2-deoxyhexos-5-ulose (together 0.18). On treatment with alkaline phosphatase further deoxy sugars were recognized and in fructose-1-phosphate G(6-deoxy-2,5-hexodiulose) was increased to a G-value of 0.4. Dephosphorylation is considered to occur mainly after OH attack at C-5 and C-1 in fructose-1-phosphate and at C-5 and C-6 in fructose-6-phosphate. Reaction mechanisms are discussed. (orig.)

  5. SLC37A1 and SLC37A2 are phosphate-linked, glucose-6-phosphate antiporters.

    Directory of Open Access Journals (Sweden)

    Chi-Jiunn Pan

    Full Text Available Blood glucose homeostasis between meals depends upon production of glucose within the endoplasmic reticulum (ER of the liver and kidney by hydrolysis of glucose-6-phosphate (G6P into glucose and phosphate (P(i. This reaction depends on coupling the G6P transporter (G6PT with glucose-6-phosphatase-α (G6Pase-α. Only one G6PT, also known as SLC37A4, has been characterized, and it acts as a P(i-linked G6P antiporter. The other three SLC37 family members, predicted to be sugar-phosphate:P(i exchangers, have not been characterized functionally. Using reconstituted proteoliposomes, we examine the antiporter activity of the other SLC37 members along with their ability to couple with G6Pase-α. G6PT- and mock-proteoliposomes are used as positive and negative controls, respectively. We show that SLC37A1 and SLC37A2 are ER-associated, P(i-linked antiporters, that can transport G6P. Unlike G6PT, neither is sensitive to chlorogenic acid, a competitive inhibitor of physiological ER G6P transport, and neither couples to G6Pase-α. We conclude that three of the four SLC37 family members are functional sugar-phosphate antiporters. However, only G6PT/SLC37A4 matches the characteristics of the physiological ER G6P transporter, suggesting the other SLC37 proteins have roles independent of blood glucose homeostasis.

  6. Tetranuclear copper(II) complexes bridged by alpha-D-glucose-1-phosphate and incorporation of sugar acids through the Cu4 core structural changes.

    Science.gov (United States)

    Kato, Merii; Sah, Ajay Kumar; Tanase, Tomoaki; Mikuriya, Masahiro

    2006-08-21

    Tetranuclear copper(II) complexes containing alpha-D-glucose-1-phosphate (alpha-D-Glc-1P), [Cu4(mu-OH){mu-(alpha-D-Glc-1P)}2(bpy)4(H2O)2]X3 [X = NO3 (1a), Cl (1b), Br (1c)], and [Cu4(mu-OH){mu-(alpha-D-Glc-1P)}2(phen)4(H2O)2](NO3)3 (2) were prepared by reacting the copper(II) salt with Na2[alpha-D-Glc-1P] in the presence of diimine ancillary ligands, and the structure of 2 was characterized by X-ray crystallography to comprise four {Cu(phen)}2+ fragments connected by the two sugar phosphate dianions in 1,3-O,O' and 1,1-O mu4-bridging fashion as well as a mu-hydroxo anion. The crystal structure of 2 involves two chemically independent complex cations in which the C2 enantiomeric structure for the trapezoidal tetracopper(II) framework is switched according to the orientation of the alpha-D-glucopyranosyl moieties. Temperature-dependent magnetic susceptibility data of 1a indicated that antiferromagnetic spin coupling is operative between the two metal ions joined by the hydroxo bridge (J = -52 cm(-1)) while antiferromagnetic interaction through the Cu-O-Cu sugar phosphate bridges is weak (J = -13 cm(-1)). Complex 1a readily reacted with carboxylic acids to afford the tetranuclear copper(II) complexes, [Cu4{mu-(alpha-D-Glc-1P)}2(mu-CA)2(bpy)4](NO3)2 [CA = CH3COO (3), o-C6H4(COO)(COOH) (4)]. Reactions with m-phenylenediacetic acid [m-C6H4(CH2COOH)2] also gave the discrete tetracopper(II) cationic complex [Cu4{mu-(alpha-D-Glc-1P)}2(mu-m-C6H4(CH2COO)(CH2COOH))2(bpy)4](NO3)2 (5a) as well as the cluster polymer formulated as {[Cu4{mu-(alpha-D-Glc-1P)}2(mu-m-C6H4(CH2COO)2)(bpy)4](NO3)2}n (5b). The tetracopper structure of 1a is converted into a symmetrical rectangular core in complexes 3, 4, and 5b, where the hydroxo bridge is dissociated and, instead, two carboxylate anions bridge another pair of Cu(II) ions in a 1,1-O monodentate fashion. The similar reactions were applied to incorporate sugar acids onto the tetranuclear copper(II) centers. Reactions of 1a with delta

  7. 21 CFR 864.7360 - Erythrocytic glucose-6-phosphate dehydrogenase assay.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Erythrocytic glucose-6-phosphate dehydrogenase assay. 864.7360 Section 864.7360 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages...

  8. Crystal Structure and Substrate Specificity of D-Galactose-6-Phosphate Isomerase Complexed with Substrates

    Science.gov (United States)

    Lee, Jung-Kul; Pan, Cheol-Ho

    2013-01-01

    D-Galactose-6-phosphate isomerase from Lactobacillus rhamnosus (LacAB; EC 5.3.1.26), which is encoded by the tagatose-6-phosphate pathway gene cluster (lacABCD), catalyzes the isomerization of D-galactose-6-phosphate to D-tagatose-6-phosphate during lactose catabolism and is used to produce rare sugars as low-calorie natural sweeteners. The crystal structures of LacAB and its complex with D-tagatose-6-phosphate revealed that LacAB is a homotetramer of LacA and LacB subunits, with a structure similar to that of ribose-5-phosphate isomerase (Rpi). Structurally, LacAB belongs to the RpiB/LacAB superfamily, having a Rossmann-like αβα sandwich fold as has been identified in pentose phosphate isomerase and hexose phosphate isomerase. In contrast to other family members, the LacB subunit also has a unique α7 helix in its C-terminus. One active site is distinctly located at the interface between LacA and LacB, whereas two active sites are present in RpiB. In the structure of the product complex, the phosphate group of D-tagatose-6-phosphate is bound to three arginine residues, including Arg-39, producing a different substrate orientation than that in RpiB, where the substrate binds at Asp-43. Due to the proximity of the Arg-134 residue and backbone Cα of the α6 helix in LacA to the last Asp-172 residue of LacB with a hydrogen bond, a six-carbon sugar-phosphate can bind in the larger pocket of LacAB, compared with RpiB. His-96 in the active site is important for ring opening and substrate orientation, and Cys-65 is essential for the isomerization activity of the enzyme. Two rare sugar substrates, D-psicose and D-ribulose, show optimal binding in the LacAB-substrate complex. These findings were supported by the results of LacA activity assays. PMID:24015281

  9. Crystal structure and substrate specificity of D-galactose-6-phosphate isomerase complexed with substrates.

    Directory of Open Access Journals (Sweden)

    Woo-Suk Jung

    Full Text Available D-Galactose-6-phosphate isomerase from Lactobacillus rhamnosus (LacAB; EC 5.3.1.26, which is encoded by the tagatose-6-phosphate pathway gene cluster (lacABCD, catalyzes the isomerization of D-galactose-6-phosphate to D-tagatose-6-phosphate during lactose catabolism and is used to produce rare sugars as low-calorie natural sweeteners. The crystal structures of LacAB and its complex with D-tagatose-6-phosphate revealed that LacAB is a homotetramer of LacA and LacB subunits, with a structure similar to that of ribose-5-phosphate isomerase (Rpi. Structurally, LacAB belongs to the RpiB/LacAB superfamily, having a Rossmann-like αβα sandwich fold as has been identified in pentose phosphate isomerase and hexose phosphate isomerase. In contrast to other family members, the LacB subunit also has a unique α7 helix in its C-terminus. One active site is distinctly located at the interface between LacA and LacB, whereas two active sites are present in RpiB. In the structure of the product complex, the phosphate group of D-tagatose-6-phosphate is bound to three arginine residues, including Arg-39, producing a different substrate orientation than that in RpiB, where the substrate binds at Asp-43. Due to the proximity of the Arg-134 residue and backbone Cα of the α6 helix in LacA to the last Asp-172 residue of LacB with a hydrogen bond, a six-carbon sugar-phosphate can bind in the larger pocket of LacAB, compared with RpiB. His-96 in the active site is important for ring opening and substrate orientation, and Cys-65 is essential for the isomerization activity of the enzyme. Two rare sugar substrates, D-psicose and D-ribulose, show optimal binding in the LacAB-substrate complex. These findings were supported by the results of LacA activity assays.

  10. Synthesis and Physicochemical Characterization of D-Tagatose-1-Phosphate: The Substrate of the Tagatose-1-Phosphate Kinase in the Phosphotransferase System-Mediated D-Tagatose Catabolic Pathway of Bacillus licheniformis.

    Science.gov (United States)

    Van der Heiden, Edwige; Delmarcelle, Michaël; Simon, Patricia; Counson, Melody; Galleni, Moreno; Freedberg, Darón I; Thompson, John; Joris, Bernard; Battistel, Marcos D

    2015-01-01

    We report the first enzymatic synthesis of D-tagatose-1-phosphate (Tag-1P) by the multicomponent phosphoenolpyruvate:sugar phosphotransferase system (PEP-PTS) present in tagatose-grown cells of Klebsiella pneumoniae. Physicochemical characterization by (31)P and (1)H nuclear magnetic resonance spectroscopy reveals that, in solution, this derivative is primarily in the pyranose form. Tag-1P was used to characterize the putative tagatose-1-phosphate kinase (TagK) of the Bacillus licheniformis PTS-mediated D-tagatose catabolic pathway (Bli-TagP). For this purpose, a soluble protein fusion was obtained with the 6 His-tagged trigger factor (TF(His6)) of Escherichia coli. The active fusion enzyme was named TagK-TF(His6). Tag-1P and D-fructose-1-phosphate are substrates for the TagK-TF(His6) enzyme, whereas the isomeric derivatives D-tagatose-6-phosphate and D-fructose-6-phosphate are inhibitors. Studies of catalytic efficiency (kcat/Km) reveal that the enzyme specificity is markedly in favor of Tag-1P as the substrate. Importantly, we show in vivo that the transfer of the phosphate moiety from PEP to the B. licheniformis tagatose-specific Enzyme II in E. coli is inefficient. The capability of the PTS general cytoplasmic components of B. subtilis, HPr and Enzyme I to restore the phosphate transfer is demonstrated. © 2015 S. Karger AG, Basel.

  11. Assessment of antioxidant activity of cane brown sugars by ABTS and DPPH radical scavenging assays: determination of their polyphenolic and volatile constituents.

    Science.gov (United States)

    Payet, Bertrand; Shum Cheong Sing, Alain; Smadja, Jacqueline

    2005-12-28

    Seven cane brown sugars (four from La Réunion, two from Mauritius, and one from France) were investigated for their polyphenol content and volatile composition in relation to their free radical scavenging capacity determined by ABTS and DPPH assays. The thin layer coated on the sugar crystal was extracted by Soxhlet extractor with dichloromethane. The volatile compounds of brown sugars were studied by GC-MS, and 43 compounds were identified. The total phenolic content of brown sugars was determined according to the Folin-Ciocalteu method. Phenolic compounds were quantified in the brown sugar extracts by LC-UV-ESI-MS. Brown sugar aqueous solutions exhibited weak free radical scavenging activity in the DPPH assay and higher antioxidant activity in the ABTS assay at relatively high concentration. The brown sugar extracts showed interesting free radical scavenging properties despite the low concentration of phenolic and volatile compounds. Sugar is a common foodstuff traditionally used for its sweetening properties, which might be accompanied by antioxidant properties arising from molecules (polyphenols, Maillard products) other than sucrose of the cane brown sugars.

  12. Diagnosis of Hunter's syndrome carriers; radioactive sulphate incorporation into fibroblasts in the presence of fructose 1-phosphate

    International Nuclear Information System (INIS)

    Toennesen, T.; Lykkelund, C.; Guettler, F.

    1982-01-01

    Mutual correction of co-cultivated fibroblasts from patients with Hunter's and Hurler's syndrome could be inhibited by either fructose 1-phosphate or mannose 6-phosphate. In the presence of fructose 1-phosphate a 50% mixture of fibroblasts from a patient with Hunter's syndrome and a normal homozygous individual showed an increased 35 S-sulphate incorporation into acid mucopolysaccharides. When fibroblast cultures from one obligate and two possible carriers of Hunter's syndrome were tested for 35 S-sulphate incorporation, the cultures showed either twice the normal 35 S-sulphate incorporation into acid mucopolysaccharides in the presence of fructose 1-phosphate or an abnormally high incorporation in the presence as well as in the absence of the sugar phosphate. (orig.)

  13. Adaptation of the Nelson-Somogyi reducing-sugar assay to a microassay using microtiter plates.

    Science.gov (United States)

    Green, F; Clausen, C A; Highley, T L

    1989-11-01

    The Nelson-Somogyi assay for reducing sugars was adapted to microtiter plates. The primary advantages of this modified assay are (i) smaller sample and reagent volumes, (ii) elimination of boiling and filtration steps, (iii) automated measurement with a dual-wavelength scanning TLC densitometer, (iv) increased range and reproducibility, and (v) automated colorimetric readings by reflectance rather than absorbance.

  14. [Safety Evaluation of Rare Sugar Syrup: Single-dose Oral Toxicity in Rats, Reverse Mutation Assay, Chromosome Aberration Assay, and Acute Non-Effect Level for Diarrhea of a Single Dose in Humans].

    Science.gov (United States)

    Yamada, Takako; Iida, Tetsuo; Takamine, Satoshi; Hayashi, Noriko; Okuma, Kazuhiro

    2015-01-01

    The safety of rare sugar syrup obtained from high-fructose corn syrup under slightly alkaline conditions was studied. Mutagenicity of rare sugar syrup was assessed by a reverse mutation assay using Salmonella typhimurium and Escherichia coli, and an in vitro chromosomal aberration assay using Chinese hamster lung cell line (CHL/IU). No mutagenicity of rare sugar syrup was detected under these experimental conditions. Oral administration of single dose (15,000 mg/kg) of rare sugar syrup to rats caused no abnormalities, suggesting no adverse effect of rare sugar syrup. In humans, the acute non-effect level of rare sugar syrup for causing diarrhea was estimated as 0.9 g/kg body weight as dry solid base in both males and females.

  15. It takes two to tango: defining an essential second active site in pyridoxal 5'-phosphate synthase.

    Directory of Open Access Journals (Sweden)

    Cyril Moccand

    Full Text Available The prevalent de novo biosynthetic pathway of vitamin B6 involves only two enzymes (Pdx1 and Pdx2 that form an ornate multisubunit complex functioning as a glutamine amidotransferase. The synthase subunit, Pdx1, utilizes ribose 5-phosphate and glyceraldehyde 3-phosphate, as well as ammonia derived from the glutaminase activity of Pdx2 to directly form the cofactor vitamer, pyridoxal 5'-phosphate. Given the fact that a single enzyme performs the majority of the chemistry behind this reaction, a complicated mechanism is anticipated. Recently, the individual steps along the reaction co-ordinate are beginning to be unraveled. In particular, the binding of the pentose substrate and the first steps of the reaction have been elucidated but it is not known if the latter part of the chemistry, involving the triose sugar, takes place in the same or a disparate site. Here, we demonstrate through the use of enzyme assays, enzyme kinetics, and mutagenesis studies that indeed a second site is involved in binding the triose sugar and moreover, is the location of the final vitamin product, pyridoxal 5'-phosphate. Furthermore, we show that product release is triggered by the presence of a PLP-dependent enzyme. Finally, we provide evidence that a single arginine residue of the C terminus of Pdx1 is responsible for coordinating co-operativity in this elaborate protein machinery.

  16. The effects of gamma irradiation on the leaching of reducing sugars, inorganic phosphate and enzymes from barley seeds during germination in water

    International Nuclear Information System (INIS)

    Kurobane, I.; Yamaguchi, H.; Sander, C.; Nilan, R.A.

    1979-01-01

    Gamma irradiation enhanced the leaching of reducing sugars from barley seeds into the water in which the seeds were shaken. Treatments prior to shaking in water, such as overnight soaking in water at 5 0 C and subsequent germination in Petri dishes for 1 or 2 days at 20 0 C, showed pronounced effects on the leaching. The highest effect, which was obtained at 500 krad irradiation, was four times higher than that of the non-irradiated control. Gamma irradiation also stimulated the leaching of inorganic phosphate and slightly that of amylases from barley seeds. When seeds from which the embryos has been removed were shaken into water, no stimulating effect on the leaching of sugars was noted. These results, combined with the irradiation effect on the embryo, suggest that the stimulated leaching of reducing sugars is due to the extreme difference in sensitivity to gamma irradiation between the production of reducing sugars in the endosperm and the development and growth of the embryo. (author)

  17. Molecular Data for a Biochemical Model of DNA Radiation Damage: Electron Impact Ionization and Dissociative Ionization of DNA Bases and Sugar-Phosphate Backbone

    Science.gov (United States)

    Dateo, Christopher E.; Fletcher, Graham D.

    2004-01-01

    As part of the database for building up a biochemical model of DNA radiation damage, electron impact ionization cross sections of sugar-phosphate backbone and DNA bases have been calculated using the improved binary-encounter dipole (iBED) model. It is found that the total ionization cross sections of C3'- and C5'-deoxyribose-phospate, two conformers of the sugar-phosphate backbone, are close to each other. Furthermore, the sum of the ionization cross sections of the separate deoxyribose and phosphate fragments is in close agreement with the C3'- and C5'-deoxyribose-phospate cross sections, differing by less than 10%. Of the four DNA bases, the ionization cross section of guanine is the largest, then in decreasing order, adenine, thymine, and cytosine. The order is in accordance with the known propensity of oxidation of the bases by ionizing radiation. Dissociative ionization (DI), a process that both ionizes and dissociates a molecule, is investigated for cytosine. The DI cross section for the formation of H and (cytosine-Hl)(+), with the cytosine ion losing H at the 1 position, is also reported. The threshold of this process is calculated to be 17.1 eV. Detailed analysis of ionization products such as in DI is important to trace the sequential steps in the biochemical process of DNA damage.

  18. Sugar analog synthesis by in vitro biocatalytic cascade: A comparison of alternative enzyme complements for dihydroxyacetone phosphate production as a precursor to rare chiral sugar synthesis.

    Science.gov (United States)

    Hartley, Carol J; French, Nigel G; Scoble, Judith A; Williams, Charlotte C; Churches, Quentin I; Frazer, Andrew R; Taylor, Matthew C; Coia, Greg; Simpson, Gregory; Turner, Nicholas J; Scott, Colin

    2017-01-01

    Carbon-carbon bond formation is one of the most challenging reactions in synthetic organic chemistry, and aldol reactions catalysed by dihydroxyacetone phosphate-dependent aldolases provide a powerful biocatalytic tool for combining C-C bond formation with the generation of two new stereo-centres, with access to all four possible stereoisomers of a compound. Dihydroxyacetone phosphate (DHAP) is unstable so the provision of DHAP for DHAP-dependent aldolases in biocatalytic processes remains complicated. Our research has investigated the efficiency of several different enzymatic cascades for the conversion of glycerol to DHAP, including characterising new candidate enzymes for some of the reaction steps. The most efficient cascade for DHAP production, comprising a one-pot four-enzyme reaction with glycerol kinase, acetate kinase, glycerophosphate oxidase and catalase, was coupled with a DHAP-dependent fructose-1,6-biphosphate aldolase enzyme to demonstrate the production of several rare chiral sugars. The limitation of batch biocatalysis for these reactions and the potential for improvement using kinetic modelling and flow biocatalysis systems is discussed.

  19. Sugar regulation of SUGAR TRANSPORTER PROTEIN 1 (STP1) expression in Arabidopsis thaliana

    Science.gov (United States)

    Cordoba, Elizabeth; Aceves-Zamudio, Denise Lizeth; Hernández-Bernal, Alma Fabiola; Ramos-Vega, Maricela; León, Patricia

    2015-01-01

    Sugars regulate the expression of many genes at the transcriptional level. In Arabidopsis thaliana, sugars induce or repress the expression of >1800 genes, including the STP1 (SUGAR TRANSPORTER PROTEIN 1) gene, which encodes an H+/monosaccharide cotransporter. STP1 transcript levels decrease more rapidly after the addition of low concentrations of sugars than the levels of other repressed genes, such as DIN6 (DARK-INDUCED 6). We found that this regulation is exerted at the transcriptional level and is initiated by phosphorylatable sugars. Interestingly, the sugar signal that modulates STP1 expression is transmitted through a HEXOKINASE 1-independent signalling pathway. Finally, analysis of the STP1 5′ regulatory region allowed us to delimit a region of 309bp that contains the cis elements implicated in the glucose regulation of STP1 expression. Putative cis-acting elements involved in this response were identified. PMID:25281700

  20. Sugar signals and the control of plant growth and development

    NARCIS (Netherlands)

    Lastdrager, Jeroen|info:eu-repo/dai/nl/357520076; Hanson, Johannes|info:eu-repo/dai/nl/304822299; Smeekens, Sjef|info:eu-repo/dai/nl/072489995

    2014-01-01

    Sugars have a central regulatory function in steering plant growth. This review focuses on information presented in the past 2 years on key players in sugar-mediated plant growth regulation, with emphasis on trehalose 6-phosphate, target of rapamycin kinase, and Snf1-related kinase 1 regulatory

  1. Recent advances in the synthesis of rare sugars using DHAP-dependent aldolases.

    Science.gov (United States)

    Li, Aimin; Cai, Li; Chen, Zhou; Wang, Mayan; Wang, Ning; Nakanishi, Hideki; Gao, Xiao-Dong; Li, Zijie

    2017-11-27

    The occurrence rates of non-communicable diseases like obesity, diabetes and hyperlipidemia have increased remarkably due to excessive consumption of a high-energy diet. Rare sugars therefore have become increasingly attractive owing to their unique nutritional properties. In the past two decades, various rare sugars have been successfully prepared guided by the "Izumoring strategy". As a valuable complement to the Izumoring approach, the controllable dihydroxyacetone phosphate (DHAP)-dependent aldolases have generally predictable regio- and stereoselectivity, which makes them powerful tools in C-C bond construction and rare sugar production. However, the main disadvantage for this group of aldolases is their strict substrate specificity toward the donor molecule DHAP, a very expensive and relatively unstable compound. Among the current methods involving DHAP, the one that couples DHAP production from inexpensive starting materials (for instance, glycerol, DL-glycerol 3-phosphate, dihydroxyacetone, and glucose) with aldol condensation appears to be the most promising. This review thus focuses on recent advances in the application of L-rhamnulose-1-phosphate aldolase (RhaD), L-fuculose-1-phosphate aldolase (FucA), and D-fructose-1,6-bisphosphate aldolase (FruA) for rare sugar synthesis in vitro and in vivo, while illustrating strategies for supplying DHAP in efficient and economical ways. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Sugar analog synthesis by in vitro biocatalytic cascade: A comparison of alternative enzyme complements for dihydroxyacetone phosphate production as a precursor to rare chiral sugar synthesis.

    Directory of Open Access Journals (Sweden)

    Carol J Hartley

    Full Text Available Carbon-carbon bond formation is one of the most challenging reactions in synthetic organic chemistry, and aldol reactions catalysed by dihydroxyacetone phosphate-dependent aldolases provide a powerful biocatalytic tool for combining C-C bond formation with the generation of two new stereo-centres, with access to all four possible stereoisomers of a compound. Dihydroxyacetone phosphate (DHAP is unstable so the provision of DHAP for DHAP-dependent aldolases in biocatalytic processes remains complicated. Our research has investigated the efficiency of several different enzymatic cascades for the conversion of glycerol to DHAP, including characterising new candidate enzymes for some of the reaction steps. The most efficient cascade for DHAP production, comprising a one-pot four-enzyme reaction with glycerol kinase, acetate kinase, glycerophosphate oxidase and catalase, was coupled with a DHAP-dependent fructose-1,6-biphosphate aldolase enzyme to demonstrate the production of several rare chiral sugars. The limitation of batch biocatalysis for these reactions and the potential for improvement using kinetic modelling and flow biocatalysis systems is discussed.

  3. Polyethylene glycol versus dual sugar assay for gastrointestinal permeability analysis: is it time to choose?

    Directory of Open Access Journals (Sweden)

    van Wijck K

    2012-07-01

    Full Text Available Kim van Wijck,1,2 Babs AFM Bessems,2 Hans MH van Eijk,2 Wim A Buurman,2 Cornelis HC Dejong,1,2 Kaatje Lenaerts1,21Top Institute Food and Nutrition, Wageningen, The Netherlands; 2Department of Surgery, NUTRIM School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Center, Maastricht, NetherlandsBackground: Increased intestinal permeability is an important measure of disease activity and prognosis. Currently, many permeability tests are available and no consensus has been reached as to which test is most suitable. The aim of this study was to compare urinary probe excretion and accuracy of a polyethylene glycol (PEG assay and dual sugar assay in a double-blinded crossover study to evaluate probe excretion and the accuracy of both tests.Methods: Gastrointestinal permeability was measured in nine volunteers using PEG 400, PEG 1500, and PEG 3350 or lactulose-rhamnose. On 4 separate days, permeability was analyzed after oral intake of placebo or indomethacin, a drug known to increase intestinal permeability. Plasma intestinal fatty acid binding protein and calprotectin levels were determined to verify compromised intestinal integrity after indomethacin consumption. Urinary samples were collected at baseline, hourly up to 5 hours after probe intake, and between 5 and 24 hours. Urinary excretion of PEG and sugars was determined using high-pressure liquid chromatography-evaporative light scattering detection and liquid chromatography-mass spectrometry, respectively.Results: Intake of indomethacin increased plasma intestinal fatty acid-binding protein and calprotectin levels, reflecting loss of intestinal integrity and inflammation. In this state of indomethacin-induced gastrointestinal compromise, urinary excretion of the three PEG probes and lactulose increased compared with placebo. Urinary PEG 400 excretion, the PEG 3350/PEG 400 ratio, and the lactulose/rhamnose ratio could accurately detect indomethacin-induced increases in

  4. Automated assay for screening the enzymatic release of reducing sugars from micronized biomass

    Directory of Open Access Journals (Sweden)

    Asther Marcel

    2010-07-01

    Full Text Available Abstract Background To reduce the production cost of bioethanol obtained from fermentation of the sugars provided by degradation of lignocellulosic biomass (i.e., second generation bioethanol, it is necessary to screen for new enzymes endowed with more efficient biomass degrading properties. This demands the set-up of high-throughput screening methods. Several methods have been devised all using microplates in the industrial SBS format. Although this size reduction and standardization has greatly improved the screening process, the published methods comprise one or more manual steps that seriously decrease throughput. Therefore, we worked to devise a screening method devoid of any manual steps. Results We describe a fully automated assay for measuring the amount of reducing sugars released by biomass-degrading enzymes from wheat-straw and spruce. The method comprises two independent and automated steps. The first step is the making of "substrate plates". It consists of filling 96-well microplates with slurry suspensions of micronized substrate which are then stored frozen until use. The second step is an enzymatic activity assay. After thawing, the substrate plates are supplemented by the robot with cell-wall degrading enzymes where necessary, and the whole process from addition of enzymes to quantification of released sugars is autonomously performed by the robot. We describe how critical parameters (amount of substrate, amount of enzyme, incubation duration and temperature were selected to fit with our specific use. The ability of this automated small-scale assay to discriminate among different enzymatic activities was validated using a set of commercial enzymes. Conclusions Using an automatic microplate sealer solved three main problems generally encountered during the set-up of methods for measuring the sugar-releasing activity of plant cell wall-degrading enzymes: throughput, automation, and evaporation losses. In its present set-up, the

  5. Cleanup and analysis of sugar phosphates in biological extracts by using solid phase extraction and anion-exchange chromatography with pulsed amperometric detection

    DEFF Research Database (Denmark)

    Smith, Hans Peter; Cohen, A.; Buttler, T.

    1998-01-01

    of Saccharomyces cerevisiae obtained by using cold methanol as quenching agent and chloroform as extraction solvent. It was shown that pretreatment of the cell extract with SPE markedly improved the quality of the liquid chromatography analysis with recoveries of the sugar phosphates close to 100%. Furthermore...

  6. Glycogen synthase activation by sugars in isolated hepatocytes.

    Science.gov (United States)

    Ciudad, C J; Carabaza, A; Bosch, F; Gòmez I Foix, A M; Guinovart, J J

    1988-07-01

    We have investigated the activation by sugars of glycogen synthase in relation to (i) phosphorylase a activity and (ii) changes in the intracellular concentration of glucose 6-phosphate and adenine nucleotides. All the sugars tested in this work present the common denominator of activating glycogen synthase. On the other hand, phosphorylase a activity is decreased by mannose and glucose, unchanged by galactose and xylitol, and increased by tagatose, glyceraldehyde, and fructose. Dihydroxyacetone exerts a biphasic effect on phosphorylase. These findings provide additional evidence proving that glycogen synthase can be activated regardless of the levels of phosphorylase a, clearly establishing that a nonsequential mechanism for the activation of glycogen synthase occurs in liver cells. The glycogen synthase activation state is related to the concentrations of glucose 6-phosphate and adenine nucleotides. In this respect, tagatose, glyceraldehyde, and fructose deplete ATP and increase AMP contents, whereas glucose, mannose, galactose, xylitol, and dihydroxyacetone do not alter the concentration of these nucleotides. In addition, all these sugars, except glyceraldehyde, increase the intracellular content of glucose 6-phosphate. The activation of glycogen synthase by sugars is reflected in decreases on both kinetic constants of the enzyme, M0.5 (for glucose 6-phosphate) and S0.5 (for UDP-glucose). We propose that hepatocyte glycogen synthase is activated by monosaccharides by a mechanism triggered by changes in glucose 6-phosphate and adenine nucleotide concentrations which have been described to modify glycogen synthase phosphatase activity. This mechanism represents a metabolite control of the sugar-induced activation of hepatocyte glycogen synthase.

  7. Substrate Specificity and Inhibitor Sensitivity of Plant UDP-Sugar Producing Pyrophosphorylases

    Directory of Open Access Journals (Sweden)

    Daniel Decker

    2017-09-01

    Full Text Available UDP-sugars are essential precursors for glycosylation reactions producing cell wall polysaccharides, sucrose, glycoproteins, glycolipids, etc. Primary mechanisms of UDP sugar formation involve the action of at least three distinct pyrophosphorylases using UTP and sugar-1-P as substrates. Here, substrate specificities of barley and Arabidopsis (two isozymes UDP-glucose pyrophosphorylases (UGPase, Arabidopsis UDP-sugar pyrophosphorylase (USPase and Arabidopsis UDP-N-acetyl glucosamine pyrophosphorylase2 (UAGPase2 were investigated using a range of sugar-1-phosphates and nucleoside-triphosphates as substrates. Whereas all the enzymes preferentially used UTP as nucleotide donor, they differed in their specificity for sugar-1-P. UGPases had high activity with D-Glc-1-P, but could also react with Fru-1-P and Fru-2-P (Km values over 10 mM. Contrary to an earlier report, their activity with Gal-1-P was extremely low. USPase reacted with a range of sugar-1-phosphates, including D-Glc-1-P, D-Gal-1-P, D-GalA-1-P (Km of 1.3 mM, β-L-Ara-1-P and α-D-Fuc-1-P (Km of 3.4 mM, but not β-L-Fuc-1-P. In contrast, UAGPase2 reacted only with D-GlcNAc-1-P, D-GalNAc-1-P (Km of 1 mM and, to some extent, D-Glc-1-P (Km of 3.2 mM. Generally, different conformations/substituents at C2, C4, and C5 of the pyranose ring of a sugar were crucial determinants of substrate specificity of a given pyrophosphorylase. Homology models of UDP-sugar binding to UGPase, USPase and UAGPase2 revealed more common amino acids for UDP binding than for sugar binding, reflecting differences in substrate specificity of these proteins. UAGPase2 was inhibited by a salicylate derivative that was earlier shown to affect UGPase and USPase activities, consistent with a common structural architecture of the three pyrophosphorylases. The results are discussed with respect to the role of the pyrophosphorylases in sugar activation for glycosylated end-products.

  8. Sphingosine-1-Phosphate (S1P) Signaling in Neural Progenitors.

    Science.gov (United States)

    Callihan, Phillip; Alqinyah, Mohammed; Hooks, Shelley B

    2018-01-01

    Sphingosine-1-phosphate (S1P) and its receptors are important in nervous system development. Reliable in vitro human model systems are needed to further define specific roles for S1P signaling in neural development. We have described S1P-regulated signaling, survival, and differentiation in a human embryonic stem cell-derived neuroepithelial progenitor cell line (hNP1) that expresses functional S1P receptors. These cells can be further differentiated to a neuronal cell type and therefore represent a good model system to study the role of S1P signaling in human neural development. The following sections describe in detail the culture and differentiation of hNP1 cells and two assays to measure S1P signaling in these cells.

  9. A Novel Complementation Assay for Quick and Specific Screen of Genes Encoding Glycerol-3-Phosphate Acyltransferases

    Directory of Open Access Journals (Sweden)

    Jie Lei

    2018-03-01

    Full Text Available The initial step in glycerolipid biosynthesis, especially in diverse allopolyploid crop species, is poorly understood, mainly due to the lack of an effective and convenient method for functional characterization of genes encoding glycerol-3-phosphate acyltransferases (GPATs catalyzing this reaction. Here we present a novel complementation assay for quick and specific characterization of GPAT-encoding genes. Its key design involves rational construction of yeast conditional lethal gat1Δgat2Δ double mutant bearing the heterologous Arabidopsis AtGPAT1 gene whose leaky expression under repressed conditions does not support any non-specific growth, thereby circumventing the false positive problem encountered with the system based on the gat1Δgat2Δ mutant harboring the native episomal GAT1 gene whose leaky expression appears to be sufficient for generating enough GPAT activities for the non-specific restoration of the mutant growth. A complementation assay developed based on this novel mutant enables quick phenotypic screen of GPAT sequences. A high degree of specificity of our assay was exemplified by its ability to differentiate effectively GPAT-encoding genes from those of other fatty acyltransferases and lipid-related sequences. Using this assay, we show that Arabidopsis AtGPAT1, AtGPAT5, and AtGPAT7 can complement the phosphatidate biosynthetic defect in the double mutants. Collectively, our assay provides a powerful tool for rapid screening, validation and optimization of GPAT sequences, aiding future engineering of the initial step of the triacylglycerol biosynthesis in oilseeds.

  10. Trehalose-6-Phosphate: connecting plant metabolism and development

    Directory of Open Access Journals (Sweden)

    Jathish ePonnu

    2011-11-01

    Full Text Available Beyond their metabolic roles, sugars can also act as messengers in signal transduction. Trehalose, a sugar found in many species of plants and animals, is a non-reducing disaccharide composed of two glucose moieties. Its synthesis in plants is a two-step process, involving the production of trehalose-6-phosphate (T6P catalyzed by TREHALOSE-6-PHOSPHATE SYNTHASE (TPS and its consecutive dephosphorylation to trehalose, catalyzed by TREHALOSE-6-PHOSPHATE PHOSPHATASE (TPP. T6P has recently emerged as an important signaling metabolite, regulating carbon assimilation and sugar status in plants. In addition, T6P has also been demonstrated to play an essential role in plant development. This review recapitulates the recent advances in our understanding the role of T6P in coordinating diverse metabolic and developmental processes.

  11. Photosynthesis Activates Plasma Membrane H+-ATPase via Sugar Accumulation.

    Science.gov (United States)

    Okumura, Masaki; Inoue, Shin-Ichiro; Kuwata, Keiko; Kinoshita, Toshinori

    2016-05-01

    Plant plasma membrane H(+)-ATPase acts as a primary transporter via proton pumping and regulates diverse physiological responses by controlling secondary solute transport, pH homeostasis, and membrane potential. Phosphorylation of the penultimate threonine and the subsequent binding of 14-3-3 proteins in the carboxyl terminus of the enzyme are required for H(+)-ATPase activation. We showed previously that photosynthesis induces phosphorylation of the penultimate threonine in the nonvascular bryophyte Marchantia polymorpha However, (1) whether this response is conserved in vascular plants and (2) the process by which photosynthesis regulates H(+)-ATPase phosphorylation at the plasma membrane remain unresolved issues. Here, we report that photosynthesis induced the phosphorylation and activation of H(+)-ATPase in Arabidopsis (Arabidopsis thaliana) leaves via sugar accumulation. Light reversibly phosphorylated leaf H(+)-ATPase, and this process was inhibited by pharmacological and genetic suppression of photosynthesis. Immunohistochemical and biochemical analyses indicated that light-induced phosphorylation of H(+)-ATPase occurred autonomously in mesophyll cells. We also show that the phosphorylation status of H(+)-ATPase and photosynthetic sugar accumulation in leaves were positively correlated and that sugar treatment promoted phosphorylation. Furthermore, light-induced phosphorylation of H(+)-ATPase was strongly suppressed in a double mutant defective in ADP-glucose pyrophosphorylase and triose phosphate/phosphate translocator (adg1-1 tpt-2); these mutations strongly inhibited endogenous sugar accumulation. Overall, we show that photosynthesis activated H(+)-ATPase via sugar production in the mesophyll cells of vascular plants. Our work provides new insight into signaling from chloroplasts to the plasma membrane ion transport mechanism. © 2016 American Society of Plant Biologists. All Rights Reserved.

  12. Nectar secretion requires sucrose phosphate synthases and the sugar transporter SWEET9.

    Science.gov (United States)

    Lin, I Winnie; Sosso, Davide; Chen, Li-Qing; Gase, Klaus; Kim, Sang-Gyu; Kessler, Danny; Klinkenberg, Peter M; Gorder, Molly K; Hou, Bi-Huei; Qu, Xiao-Qing; Carter, Clay J; Baldwin, Ian T; Frommer, Wolf B

    2014-04-24

    Angiosperms developed floral nectaries that reward pollinating insects. Although nectar function and composition have been characterized, the mechanism of nectar secretion has remained unclear. Here we identify SWEET9 as a nectary-specific sugar transporter in three eudicot species: Arabidopsis thaliana, Brassica rapa (extrastaminal nectaries) and Nicotiana attenuata (gynoecial nectaries). We show that SWEET9 is essential for nectar production and can function as an efflux transporter. We also show that sucrose phosphate synthase genes, encoding key enzymes for sucrose biosynthesis, are highly expressed in nectaries and that their expression is also essential for nectar secretion. Together these data are consistent with a model in which sucrose is synthesized in the nectary parenchyma and subsequently secreted into the extracellular space via SWEET9, where sucrose is hydrolysed by an apoplasmic invertase to produce a mixture of sucrose, glucose and fructose. The recruitment of SWEET9 for sucrose export may have been a key innovation, and could have coincided with the evolution of core eudicots and contributed to the evolution of nectar secretion to reward pollinators.

  13. Conformation and functioning of tRNAs: cross-linked tRNAs as substrate for tRNA nucleotidyl-transferase and aminoacyl synthetases

    International Nuclear Information System (INIS)

    Carre, D.S.; Thomas, G.; Favre, A.

    1974-01-01

    The behavior of mixed E. coli tRNAs ''cross-linked'' by irradiation with near ultraviolet light (310-400 nm) has been compared to that of the intact molecules in two enzymatic processes. No change in the rate and extent of the repair of the pCpCpA 3' terminus of tRNA by purified E. coli tRNA nucleotidyltransferase can be detected. In contrast, complex data were obtained in the acylation reaction. They can be understood using other tRNA specific modifications as well as our present knowledge of E. coli tRNA sequences and rare base content [fr

  14. Rapid 2,2'-bicinchoninic-based xylanase assay compatible with high throughput screening

    Science.gov (United States)

    William R. Kenealy; Thomas W. Jeffries

    2003-01-01

    High-throughput screening requires simple assays that give reliable quantitative results. A microplate assay was developed for reducing sugar analysis that uses a 2,2'-bicinchoninic-based protein reagent. Endo-1,4-â-D-xylanase activity against oat spelt xylan was detected at activities of 0.002 to 0.011 IU ml−1. The assay is linear for sugar...

  15. Polyethylene glycol versus dual sugar assay for gastrointestinal permeability analysis: is it time to choose?

    Science.gov (United States)

    van Wijck, Kim; Bessems, Babs Afm; van Eijk, Hans Mh; Buurman, Wim A; Dejong, Cornelis Hc; Lenaerts, Kaatje

    2012-01-01

    Increased intestinal permeability is an important measure of disease activity and prognosis. Currently, many permeability tests are available and no consensus has been reached as to which test is most suitable. The aim of this study was to compare urinary probe excretion and accuracy of a polyethylene glycol (PEG) assay and dual sugar assay in a double-blinded crossover study to evaluate probe excretion and the accuracy of both tests. Gastrointestinal permeability was measured in nine volunteers using PEG 400, PEG 1500, and PEG 3350 or lactulose-rhamnose. On 4 separate days, permeability was analyzed after oral intake of placebo or indomethacin, a drug known to increase intestinal permeability. Plasma intestinal fatty acid binding protein and calprotectin levels were determined to verify compromised intestinal integrity after indomethacin consumption. Urinary samples were collected at baseline, hourly up to 5 hours after probe intake, and between 5 and 24 hours. Urinary excretion of PEG and sugars was determined using high-pressure liquid chromatography-evaporative light scattering detection and liquid chromatography-mass spectrometry, respectively. Intake of indomethacin increased plasma intestinal fatty acid-binding protein and calprotectin levels, reflecting loss of intestinal integrity and inflammation. In this state of indomethacin-induced gastrointestinal compromise, urinary excretion of the three PEG probes and lactulose increased compared with placebo. Urinary PEG 400 excretion, the PEG 3350/PEG 400 ratio, and the lactulose/rhamnose ratio could accurately detect indomethacin-induced increases in gastrointestinal permeability, especially within 2 hours of probe intake. Hourly urinary excretion and diagnostic accuracy of PEG and sugar probes show high concordance for detection of indomethacin-induced increases in gastrointestinal permeability. This comparative study improves our knowledge of permeability analysis in man by providing a clear overview of both

  16. Characterization, cloning and sequencing of a thermostable endo-(1, 3-1, 4) beta-glucanase-encoding gene from an alkalophilic Bacillus-brevis

    CSIR Research Space (South Africa)

    Louw, M

    1993-01-01

    Full Text Available - zyme that produced 1 ~tmol reducing sugar calculated as glucose per minute under the conditions of assay. Bacterial strains, growth media and vectors. The Escherichia coli host strain for the original cloning experiment... the gels were washed in phosphate buffer, pH 6.3 (Beguin 1983). The bands of enzyme activity were detected by staining the lichen- an/PAGE gel with Congo red. Restriction mapping and nucleotide sequencing. Restriction en...

  17. The Determination of Sugars by Chromatographic Method

    OpenAIRE

    Sumartini, Sri; Kantasubrata, Julia

    1992-01-01

    Experiments have been carried out to analyse sugars using TLC and HPLC methods, In the TLC method, separation of sugars was performed on silica plates impregnated with monosodium phosphate and using mixture of ethylacettuel pyridinde/water as an eluent. Whilst in the HPLC method, the use of three column types i.e. diol, RP-18 and modified silica column were tested. The results showed that TLC method was able to measure three sugars i:e. sucrose, glucose and fructose with standard deviations o...

  18. A high-performance liquid chromatography-based radiometric assay for sucrose-phosphate synthase and other UDP-glucose requiring enzymes

    International Nuclear Information System (INIS)

    Salvucci, M.E.; Crafts-Brandner, S.J.

    1991-01-01

    A method for product analysis that eliminates a problematic step in the radiometric sucrose-phosphate synthase assay is described. The method uses chromatography on a boronate-derivatized high-performance liquid chromatography column to separate the labeled product, [14C]sucrose phosphate, from unreacted uridine 5'-diphosphate-[14C]glucose (UDP-Glc). Direct separation of these compounds eliminates the need for treatment of the reaction mixtures with alkaline phosphatase, thereby avoiding the problem of high background caused by contaminating phosphodiesterase activity in alkaline phosphatase preparations. The method presented in this paper can be applied to many UDP-Glc requiring enzymes; here the authors show its use for determining the activities of sucrose-phosphate synthase, sucrose synthase, and uridine diphosphate-glucose pyrophosphorylase in plant extracts

  19. Role of Phosphate Transport System Component PstB1 in Phosphate Internalization by Nostoc punctiforme.

    Science.gov (United States)

    Hudek, L; Premachandra, D; Webster, W A J; Bräu, L

    2016-11-01

    In bacteria, limited phosphate availability promotes the synthesis of active uptake systems, such as the Pst phosphate transport system. To understand the mechanisms that facilitate phosphate accumulation in the cyanobacterium Nostoc punctiforme, phosphate transport systems were identified, revealing a redundancy of Pst phosphate uptake systems that exists across three distinct operons. Four separate PstB system components were identified. pstB1 was determined to be a suitable target for creating phenotypic mutations that could result in the accumulation of excessive levels of phosphate through its overexpression or in a reduction of the capacity to accumulate phosphate through its deletion. Using quantitative real-time PCR (qPCR), it was determined that pstB1 mRNA levels increased significantly over 64 h in cells cultured in 0 mM added phosphate and decreased significantly in cells exposed to high (12.8 mM) phosphate concentrations compared to the level in cells cultured under normal (0.8 mM) conditions. Possible compensation for the loss of PstB1 was observed when pstB2, pstB3, and pstB4 mRNA levels increased, particularly in cells starved of phosphate. The overexpression of pstB1 increased phosphate uptake by N. punctiforme and was shown to functionally complement the loss of PstB in E. coli PstB knockout (PstB - ) mutants. The knockout of pstB1 in N. punctiforme did not have a significant effect on cellular phosphate accumulation or growth for the most part, which is attributed to the compensation for the loss of PstB1 by alterations in the pstB2, pstB3, and pstB4 mRNA levels. This study provides novel in vivo evidence that PstB1 plays a functional role in phosphate uptake in N. punctiforme IMPORTANCE: Cyanobacteria have been evolving over 3.5 billion years and have become highly adept at growing under limiting nutrient levels. Phosphate is crucial for the survival and prosperity of all organisms. In bacteria, limited phosphate availability promotes the

  20. Pentose phosphates in nucleoside interconversion and catabolism.

    Science.gov (United States)

    Tozzi, Maria G; Camici, Marcella; Mascia, Laura; Sgarrella, Francesco; Ipata, Piero L

    2006-03-01

    Ribose phosphates are either synthesized through the oxidative branch of the pentose phosphate pathway, or are supplied by nucleoside phosphorylases. The two main pentose phosphates, ribose-5-phosphate and ribose-1-phosphate, are readily interconverted by the action of phosphopentomutase. Ribose-5-phosphate is the direct precursor of 5-phosphoribosyl-1-pyrophosphate, for both de novo and 'salvage' synthesis of nucleotides. Phosphorolysis of deoxyribonucleosides is the main source of deoxyribose phosphates, which are interconvertible, through the action of phosphopentomutase. The pentose moiety of all nucleosides can serve as a carbon and energy source. During the past decade, extensive advances have been made in elucidating the pathways by which the pentose phosphates, arising from nucleoside phosphorolysis, are either recycled, without opening of their furanosidic ring, or catabolized as a carbon and energy source. We review herein the experimental knowledge on the molecular mechanisms by which (a) ribose-1-phosphate, produced by purine nucleoside phosphorylase acting catabolically, is either anabolized for pyrimidine salvage and 5-fluorouracil activation, with uridine phosphorylase acting anabolically, or recycled for nucleoside and base interconversion; (b) the nucleosides can be regarded, both in bacteria and in eukaryotic cells, as carriers of sugars, that are made available though the action of nucleoside phosphorylases. In bacteria, catabolism of nucleosides, when suitable carbon and energy sources are not available, is accomplished by a battery of nucleoside transporters and of inducible catabolic enzymes for purine and pyrimidine nucleosides and for pentose phosphates. In eukaryotic cells, the modulation of pentose phosphate production by nucleoside catabolism seems to be affected by developmental and physiological factors on enzyme levels.

  1. Early response of plant cell to carbon deprivation: in vivo 31P-NMR spectroscopy shows a quasi-instantaneous disruption on cytosolic sugars, phosphorylated intermediates of energy metabolism, phosphate partitioning, and intracellular pHs.

    Science.gov (United States)

    Gout, Elisabeth; Bligny, Richard; Douce, Roland; Boisson, Anne-Marie; Rivasseau, Corinne

    2011-01-01

    • In plant cells, sugar starvation triggers a cascade of effects at the scale of 1-2 days. However, very early metabolic response has not yet been investigated. • Soluble phosphorus (P) compounds and intracellular pHs were analysed each 2.5 min intervals in heterotrophic sycamore (Acer pseudoplatanus) cells using in vivo phosphorus nuclear magnetic resonance ((31)P-NMR). • Upon external-sugar withdrawal, the glucose 6-P concentration dropped in the cytosol, but not in plastids. The released inorganic phosphate (Pi) accumulated transiently in the cytosol before influx into the vacuole; nucleotide triphosphate concentration doubled, intracellular pH increased and cell respiration decreased. It was deduced that the cytosolic free-sugar concentration was low, corresponding to only 0.5 mM sucrose in sugar-supplied cells. • The release of sugar from the vacuole and from plastids is insufficient to fully sustain the cell metabolism during starvation, particularly in the very short term. Similarly to Pi-starvation, the cell's first response to sugar starvation occurs in the cytosol and is of a metabolic nature. Unlike the cytoplasm, cytosolic homeostasis is not maintained during starvation. The important metabolic changes following cytosolic sugar exhaustion deliver early endogenous signals that may contribute to trigger rescue metabolism. © The Authors (2010). Journal compilation © New Phytologist Trust (2010).

  2. Complex formation of uranium(VI) with fructose and glucose phosphates

    International Nuclear Information System (INIS)

    Koban, A.; Geipel, G.; Bernhard, G.; Fanghaenel, T.

    2002-01-01

    The uptake of heavy metals into plants is commonly quantified by the soil-plant transfer factor. Up to now little is known about the chemical speciation of actinides in plants. To compare the obtained spectroscopic data of uranium complexes in plants with model compounds, we investigate the complexation of uranium with relevant bioligands of various functionalities. A very important class of ligands consists of phosphate esters, which serve as phosphate group and energy transmitters as well as energy storage media in biological systems. Heavy metal ions bound to the phosphate esters can be transported into living cells and then deposited. Therefore, in our study we present the results of uranium complexation with glucose-6-phosphate (G6P), and fructose-6-phosphate (F6P) obtained by time-resolved laser-induced fluorescence spectroscopy (TRLFS). The experiments were performed at a fixed uranyl concentration (10 -5 M) as a function of the ligand concentrations (10 -5 to 10 -3 M) in a pH range from 2 to 4.5. For the glucose phosphate system we observed, using increasing ligand concentrations, a decrease in the fluorescence intensity and a small red shift of the emission bands. From this we conclude that the complexed uranyl glucose phosphate species show only minor or no fluorescence properties. The TRLFS spectra of the glucose phosphate samples indicated the presence of a single species with fluorescence properties. This species has a lifetime of approximately 1.5 μs and was identified as the free uranyl ion. An opposite phenomenon was observed for the fructose phosphate system: there was no decrease in fluorescence intensity. However, a strong red shift of the spectra was observed, illustrating the fluorescence properties of the uranyl fructose phosphate complex. The TRLFS spectra of the fructose phosphate system showed a second lifetime ( 2 2+ UO 2 (lig) x (2-y)+ + y H + (lig = sugar phosphate). Applying the mass action law and transformation to the logarithmic

  3. Phosphate analysis of natural sausage casings preserved in brines with phosphate additives as inactivating agent - Method validation.

    Science.gov (United States)

    Wijnker, J J; Tjeerdsma-van Bokhoven, J L M; Veldhuizen, E J A

    2009-01-01

    Certain phosphates have been identified as suitable additives for the improvement of the microbial and mechanical properties of processed natural sausage casings. When mixed with NaCl (sodium chloride) and used under specific treatment and storage conditions, these phosphates are found to prevent the spread of foot-and-mouth disease and classical swine fever via treated casings. The commercially available Quantichrom™ phosphate assay kit has been evaluated as to whether it can serve as a reliable and low-tech method for routine analysis of casings treated with phosphate. The outcome of this study indicates that this particular assay kit has sufficient sensitivity to qualitatively determine the presence of phosphate in treated casings without interference of naturally occurring phosphate in salt used for brines in which casings are preserved.

  4. Assay system

    International Nuclear Information System (INIS)

    Patzke, J.B.; Rosenberg, B.J.

    1984-01-01

    The accuracy of assays for monitoring concentrations of basic drugs in biological fluids containing a 1 -acid glycoproteins, such as blood (serum or plasma), is improved by the addition of certain organic phosphate compounds to minimize the ''protein effect.'' Kits containing the elements of the invention are also disclosed

  5. Influence of insulin on heat (450) protection by hexose sugars

    International Nuclear Information System (INIS)

    Sandifer, L.; Nagle, W.A.; Henle, K.J.; Moss, A.J. Jr.

    1987-01-01

    Treatment of cultured cells with 100mM D-glucose and D-galactose confers protection against hyperthermia-induced cell death, but the mechanism is not known. The authors measured changes in cell survival and altered levels of intracellular sugar metabolites in Chinese hamster fibroblast (V79) cells. Cells were incubated at 37 0 for 1 or 5 hours prior to a 45 0 heating in balanced salts solution (BSS) with 2mM glutamine and varying concentrations of sugars in the presence and absence of insulin (10 gm/ml). Cells incubated at all sugar concentrations (5-125mM) with insulin showed a more rapid increase in survival: after 17 min. at 45 0 the survival with 125mM sugar plus insulin yielded a 4 fold increase after a 1 or 5 hour incubation. Longer incubation times were required for increased survival in the absence of insulin. The authors also observed increased survival, relative to cells heated in complete medium, for cells incubated in BSS with 2mM glutamine and no sugar. This suggests that glutamine metabolism may lead to an increase in cell heat resistance. These survival results will be related to intracellular changes in sugar metabolites, principally sugar phosphates

  6. The DNA and RNA sugar-phosphate backbone emerges as the key player. An overview of quantum-chemical, structural biology and simulation studies

    Czech Academy of Sciences Publication Activity Database

    Šponer, Jiří; Mládek, Arnošt; Šponer, Judit E.; Svozil, Daniel; Zgarbová, M.; Banáš, Pavel; Jurečka, P.; Otyepka, M.

    2012-01-01

    Roč. 14, č. 44 (2012), s. 15257-15277 ISSN 1463-9076 R&D Projects: GA ČR(CZ) GD203/09/H046; GA ČR(CZ) GAP208/10/2302; GA ČR(CZ) GAP208/11/1822; GA ČR(CZ) GAP208/12/1878; GA ČR(CZ) GA203/09/1476; GA ČR(CZ) GBP305/12/G034 Institutional research plan: CEZ:AV0Z50040702 Keywords : DNA * RNA * sugar-phosphate backbone Subject RIV: BO - Biophysics Impact factor: 3.829, year: 2012

  7. Trehalose-6-phosphate synthesis controls yeast gluconeogenesis downstream and independent of SNF1.

    Science.gov (United States)

    Deroover, Sofie; Ghillebert, Ruben; Broeckx, Tom; Winderickx, Joris; Rolland, Filip

    2016-06-01

    Trehalose-6-P (T6P), an intermediate of trehalose biosynthesis, was identified as an important regulator of yeast sugar metabolism and signaling. tps1Δ mutants, deficient in T6P synthesis (TPS), are unable to grow on rapidly fermentable medium with uncontrolled influx in glycolysis, depletion of ATP and accumulation of sugar phosphates. However, the exact molecular mechanisms involved are not fully understood. We show that SNF1 deletion restores the tps1Δ growth defect on glucose, suggesting that lack of TPS hampers inactivation of SNF1 or SNF1-regulated processes. In addition to alternative, non-fermentable carbon metabolism, SNF1 controls two major processes: respiration and gluconeogenesis. The tps1Δ defect appears to be specifically associated with deficient inhibition of gluconeogenesis, indicating more downstream effects. Consistently, Snf1 dephosphorylation and inactivation on glucose medium are not affected, as confirmed with an in vivo Snf1 activity reporter. Detailed analysis shows that gluconeogenic Pck1 and Fbp1 expression, protein levels and activity are not repressed upon glucose addition to tps1Δ cells, suggesting a link between the metabolic defect and persistent gluconeogenesis. While SNF1 is essential for induction of gluconeogenesis, T6P/TPS is required for inactivation of gluconeogenesis in the presence of glucose, downstream and independent of SNF1 activity and the Cat8 and Sip4 transcription factors. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  8. Loss of peroxisomes causes oxygen insensitivity of the histochemical assay of glucose-6-phosphate dehydrogenase activity to detect cancer cells

    NARCIS (Netherlands)

    Frederiks, Wilma M.; Vreeling-Sindelárová, Heleen; van Noorden, Cornelis J. F.

    2007-01-01

    Oxygen insensitivity of carcinoma cells and oxygen sensitivity of non-cancer cells in the histochemical assay of glucose-6-phosphate dehydrogenase (G6PD) enables detection of carcinoma cells in unfixed cell smears or cryostat sections of biopsies. The metabolic background of oxygen insensitivity is

  9. Thermodynamics of the hydrolysis reactions of α-D-galactose 1-phosphate, sn-glycerol 3-phosphate, 4-nitrophenyl phosphate, phosphocreatine, and 3-phospho-D-glycerate

    International Nuclear Information System (INIS)

    Goldberg, Robert N.; Lang, Brian E.; Lo, Catherine; Ross, David J.; Tewari, Yadu B.

    2009-01-01

    Microcalorimetry, high-performance liquid chromatography (h.p.l.c.), and an enzymatic assay have been used to conduct a thermodynamic investigation of five phosphate hydrolysis reactions: {α-D-galactose 1-phosphate(aq) + H 2 O(l) = D-galactose(aq) + orthophosphate(aq)} (1), {sn-glycerol 3-phosphate(aq) + H 2 O(l) = glycerol(aq) + orthophosphate(aq)} (2), {4-nitrophenyl phosphate(aq) + H 2 O(l) = 4-nitrophenol(aq) + orthophosphate(aq)} (3), {phosphocreatine(aq) + H 2 O(l) = creatine(aq) + orthophosphate(aq)} (4), and {3-phospho-D-glycerate(aq) + H 2 O(l) = D-glycerate(aq) + orthophosphate(aq)} (5). Calorimetrically determined enthalpies of reaction Δ r H(cal) were measured for reactions (1)-(5) and the apparent equilibrium constant K' was measured for reaction (2). The pKs and standard enthalpies of reaction Δ r H 0 for the H + and Mg 2+ binding reactions of the reactants and products in the aforementioned reactions were obtained either from the literature or by estimation. A chemical equilibrium model was then used to calculate standard equilibrium constants K and standard enthalpies of reaction Δ r H 0 for chemical reference reactions that correspond to the overall biochemical reactions that were studied experimentally. Property values from the literature and thermodynamic network calculations were used to obtain values of the equilibrium constants for the chemical reference reactions that correspond to the overall biochemical reactions (1). These values were compared with other results from the literature and also correlated with structural features. The results obtained in this study can be used in the chemical equilibrium model to calculate values of K', the standard apparent Gibbs free energy changes Δ r G '0 , the standard apparent enthalpy changes Δ r H '0 , changes in binding of the proton Δ r N(H + ), and the position of equilibrium for the overall biochemical reactions considered in this study over a reasonably wide range of temperature, pH, p

  10. High-sugar intake does not exacerbate metabolic abnormalities or cardiac dysfunction in genetic cardiomyopathy.

    Science.gov (United States)

    Hecker, Peter A; Galvao, Tatiana F; O'Shea, Karen M; Brown, Bethany H; Henderson, Reney; Riggle, Heather; Gupte, Sachin A; Stanley, William C

    2012-05-01

    A high-sugar intake increases heart disease risk in humans. In animals, sugar intake accelerates heart failure development by increased reactive oxygen species (ROS). Glucose-6-phosphate dehydrogenase (G6PD) can fuel ROS production by providing reduced nicotinamide adenine dinucleotide phosphate (NADPH) for superoxide generation by NADPH oxidase. Conversely, G6PD also facilitates ROS scavenging using the glutathione pathway. We hypothesized that a high-sugar intake would increase flux through G6PD to increase myocardial NADPH and ROS and accelerate cardiac dysfunction and death. Six-week-old TO-2 hamsters, a non-hypertensive model of genetic cardiomyopathy caused by a δ-sarcoglycan mutation, were fed a long-term diet of high starch or high sugar (57% of energy from sucrose plus fructose). After 24 wk, the δ-sarcoglycan-deficient animals displayed expected decreases in survival and cardiac function associated with cardiomyopathy (ejection fraction: control 68.7 ± 4.5%, TO-2 starch 46.1 ± 3.7%, P sugar 58.0 ± 4.2%, NS, versus TO-2 starch or control; median survival: TO-2 starch 278 d, TO-2 sugar 318 d, P = 0.133). Although the high-sugar intake was expected to exacerbate cardiomyopathy, surprisingly, there was no further decrease in ejection fraction or survival with high sugar compared with starch in cardiomyopathic animals. Cardiomyopathic animals had systemic and cardiac metabolic abnormalities (increased serum lipids and glucose and decreased myocardial oxidative enzymes) that were unaffected by diet. The high-sugar intake increased myocardial superoxide, but NADPH and lipid peroxidation were unaffected. A sugar-enriched diet did not exacerbate ventricular function, metabolic abnormalities, or survival in heart failure despite an increase in superoxide production. Copyright © 2012 Elsevier Inc. All rights reserved.

  11. Influence of protonation or alkylation of the phosphate group on the e. s. r. spectra and on the rate of phosphate elimination from 2-methoxyethyl phosphate 2-yl radicals. [. gamma. rays

    Energy Technology Data Exchange (ETDEWEB)

    Behrens, G; Koltzenburg, G; Ritter, A; Schulte-Frohlinde, D [Max-Planck-Institut fuer Kohlenforschung, Muelheim an der Ruhr (Germany, F.R.). Inst. fuer Strahlenchemie

    1978-02-01

    The e.s.r. spectra of l-yl, 2-yl and 3'-yl methoxethyl phosphate radicals derived from CH/sub 3/OCH/sub 2/CH/sub 2/-OPO/sub 3/H/sub 2/ by hydrogen abstraction have been measured in aqueous solutions and the hyperfine constants determined. The coupling constants vary strongly with protonation or alkylation of the phosphate group. The 2-yl radicals eliminate phosphate. The rate-constants for the elimination (ksub(e)) have been estimated by e.s.r. measurements and by product studies as a function of pH using /sup 60/Co ..gamma..-radiolysis. The ksub(e) values vary from approximately 0.3 s/sup -1/ for the CH/sub 3/OCHCH/sub 2/OPO/sub 3//sup - -/ radical and approximately 10/sup 3/s/sup -1/ for CH/sub 3/OCHCH/sub 2/OPO/sub 3/H/sup -/, to approximately 3 x 10/sup 6/s/sup -1/ for CH/sub 3/OCHCH/sub 2/OPO/sub 3/H/sub 2/. Alkylation of the phosphate group increased the elimination rate-constant to a similar extent as protonation. The results support a recent mechanism which described the OH-radical-induced single-strand breaks of DNA in aqueous solution starting from the C-4' radical of the sugar moiety. It is further concluded the C-4' radical of DNA eliminates the 3'-phosphate group faster than the 5'-phosphate group.

  12. 2-O-α-D-glucosylglycerol phosphorylase from Bacillus selenitireducens MLS10 possessing hydrolytic activity on β-D-glucose 1-phosphate.

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    Takanori Nihira

    Full Text Available The glycoside hydrolase family (GH 65 is a family of inverting phosphorylases that act on α-glucosides. A GH65 protein (Bsel_2816 from Bacillus selenitireducens MLS10 exhibited inorganic phosphate (Pi-dependent hydrolysis of kojibiose at the rate of 0.43 s(-1. No carbohydrate acted as acceptor for the reverse phosphorolysis using β-D-glucose 1-phosphate (βGlc1P as donor. During the search for a suitable acceptor, we found that Bsel_2816 possessed hydrolytic activity on βGlc1P with a k cat of 2.8 s(-1; moreover, such significant hydrolytic activity on sugar 1-phosphate had not been reported for any inverting phosphorylase. The H2 (18O incorporation experiment and the anomeric analysis during the hydrolysis of βGlc1P revealed that the hydrolysis was due to the glucosyl-transferring reaction to a water molecule and not a phosphatase-type reaction. Glycerol was found to be the best acceptor to generate 2-O-α-D-glucosylglycerol (GG at the rate of 180 s(-1. Bsel_2816 phosphorolyzed GG through sequential Bi-Bi mechanism with a k cat of 95 s(-1. We propose 2-O-α-D-glucopyranosylglycerol: phosphate β-D-glucosyltransferase as the systematic name and 2-O-α-D-glucosylglycerol phosphorylase as the short name for Bsel_2816. This is the first report describing a phosphorylase that utilizes polyols, and not carbohydrates, as suitable acceptor substrates.

  13. Peroxisome Proliferator-Activated Receptor γ Regulates the Expression of Lipid Phosphate Phosphohydrolase 1 in Human Vascular Endothelial Cells

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    Yazi Huang

    2014-01-01

    Full Text Available Lipid phosphate phosphohydrolase 1 (LPP1, a membrane ectophosphohydrolase regulating the availability of bioactive lipid phosphates, plays important roles in cellular signaling and physiological processes such as angiogenesis and endothelial migration. However, the regulated expression of LPP1 remains largely unknown. Here, we aimed to examine a role of peroxisome proliferator-activated receptor γ (PPARγ in the transcriptional control of LPP1 gene expression. In human umbilical vein endothelial cells (HUVECs, quantitative reverse transcriptase polymerase chain reaction (qRT-PCR demonstrated that activation of PPARγ increased the mRNA level of LPP1. Chromatin immunoprecipitation assay showed that PPARγ binds to the putative PPAR-responsive elements (PPREs within the 5′-flanking region of the human LPP1 gene. Genomic fragment containing 1.7-kilobase of the promoter region was cloned by using PCR. The luciferase reporter assays demonstrated that overexpression of PPARγ and rosiglitazone, a specific ligand for PPARγ, could significantly upregulate the reporter activity. However, site-directed mutagenesis of the PPRE motif abolished the induction. In conclusion, our results demonstrated that PPARγ transcriptionally activated the expression of LPP1 gene in ECs, suggesting a potential role of PPARγ in the metabolism of phospholipids.

  14. Distinct generation, pharmacology, and distribution of sphingosine 1-phosphate and dihydro-sphingosine 1-phosphate in human neural progenitor cells

    Science.gov (United States)

    In-vivo and in-vitro studies suggest a crucial role for Sphingosine 1-phosphate (S1P) and its receptors in the development of the nervous system. Dihydrosphingosine 1-phosphate (dhS1P), a reduced form of S1P, is an active ligand at S1P receptors, but the pharmacology and physiology of dhS1P has not...

  15. Bioconversion of lignocellulose-derived sugars to ethanol by engineered Saccharomyces cerevisiae.

    Science.gov (United States)

    Madhavan, Anjali; Srivastava, Aradhana; Kondo, Akihiko; Bisaria, Virendra S

    2012-03-01

    Lignocellulosic biomass from agricultural and agro-industrial residues represents one of the most important renewable resources that can be utilized for the biological production of ethanol. The yeast Saccharomyces cerevisiae is widely used for the commercial production of bioethanol from sucrose or starch-derived glucose. While glucose and other hexose sugars like galactose and mannose can be fermented to ethanol by S. cerevisiae, the major pentose sugars D-xylose and L-arabinose remain unutilized. Nevertheless, D-xylulose, the keto isomer of xylose, can be fermented slowly by the yeast and thus, the incorporation of functional routes for the conversion of xylose and arabinose to xylulose or xylulose-5-phosphate in Saccharomyces cerevisiae can help to improve the ethanol productivity and make the fermentation process more cost-effective. Other crucial bottlenecks in pentose fermentation include low activity of the pentose phosphate pathway enzymes and competitive inhibition of xylose and arabinose transport into the cell cytoplasm by glucose and other hexose sugars. Along with a brief introduction of the pretreatment of lignocellulose and detoxification of the hydrolysate, this review provides an updated overview of (a) the key steps involved in the uptake and metabolism of the hexose sugars: glucose, galactose, and mannose, together with the pentose sugars: xylose and arabinose, (b) various factors that play a major role in the efficient fermentation of pentose sugars along with hexose sugars, and (c) the approaches used to overcome the metabolic constraints in the production of bioethanol from lignocellulose-derived sugars by developing recombinant S. cerevisiae strains.

  16. Impact of Phosphate, Potassium, Yeast Extract, and Trace Metals on Chitosan and Metabolite Production by Mucor indicus.

    Science.gov (United States)

    Safaei, Zahra; Karimi, Keikhosro; Zamani, Akram

    2016-08-30

    In this study the effects of phosphate, potassium, yeast extract, and trace metals on the growth of Mucor indicus and chitosan, chitin, and metabolite production by the fungus were investigated. Maximum yield of chitosan (0.32 g/g cell wall) was obtained in a phosphate-free medium. Reversely, cell growth and ethanol formation by the fungus were positively affected in the presence of phosphate. In a phosphate-free medium, the highest chitosan content (0.42 g/g cell wall) and cell growth (0.66 g/g sugar) were obtained at 2.5 g/L of KOH. Potassium concentration had no significant effect on ethanol and glycerol yields. The presence of trace metals significantly increased the chitosan yield at an optimal phosphate and potassium concentration (0.50 g/g cell wall). By contrast, production of ethanol by the fungus was negatively affected (0.33 g/g sugars). A remarkable increase in chitin and decrease in chitosan were observed in the absence of yeast extract and concentrations lower than 2 g/L. The maximum chitosan yield of 51% cell wall was obtained at 5 g/L of yeast extract when the medium contained no phosphate, 2.5 g/L KOH, and 1 mL/L trace metal solution.

  17. Trehalose-6-phosphate and SnRK1 kinases in plant development and signaling: the emerging picture

    Directory of Open Access Journals (Sweden)

    Sonia eGazzarrini

    2014-04-01

    Full Text Available Carbohydrates, or sugars, regulate various aspects of plant growth through modulation of cell division and expansion. Besides playing essential roles as sources of energy for growth and as structural components of cells, carbohydrates also regulate the timing of expression of developmental programs. The disaccharide trehalose is used as an energy source, as a storage and transport molecule for glucose, and as a stress-responsive compound important for cellular protection during stress in all kingdoms. Trehalose, however, is found in very low amounts in most plants, pointing to a signaling over metabolic role for this non-reducing disaccharide. In the last decade, trehalose-6-phosphate (T6P, an intermediate in trehalose metabolism, has been shown to regulate embryonic and vegetative development, flowering time, meristem determinacy and cell fate specification in plants. T6P acts as a global regulator of metabolism and transcription promoting plant growth and triggering developmental phase transitions in response to sugar availability. Among the T6P targets are members of the Sucrose-non-fermenting1-Related Kinase1 (SnRK1 family, which are sensors of energy availability and inhibit plant growth and development during metabolic stress to maintain energy homeostasis. In this review, we will discuss the opposite roles of the sugar metabolite T6P and the SnRK1 kinases in the regulation of developmental phase transitions in response to carbohydrate levels. We will focus on how these two global regulators of metabolic processes integrate environmental cues and interact with hormonal signaling pathways to modulate plant development.

  18. Co-precipitation of phosphate and iron limits mitochondrial phosphate availability in Saccharomyces cerevisiae lacking the yeast frataxin homologue (YFH1).

    Science.gov (United States)

    Seguin, Alexandra; Santos, Renata; Pain, Debkumar; Dancis, Andrew; Camadro, Jean-Michel; Lesuisse, Emmanuel

    2011-02-25

    Saccharomyces cerevisiae cells lacking the yeast frataxin homologue (Δyfh1) accumulate iron in the mitochondria in the form of nanoparticles of ferric phosphate. The phosphate content of Δyfh1 mitochondria was higher than that of wild-type mitochondria, but the proportion of mitochondrial phosphate that was soluble was much lower in Δyfh1 cells. The rates of phosphate and iron uptake in vitro by isolated mitochondria were higher for Δyfh1 than wild-type mitochondria, and a significant proportion of the phosphate and iron rapidly became insoluble in the mitochondrial matrix, suggesting co-precipitation of these species after oxidation of iron by oxygen. Increasing the amount of phosphate in the medium decreased the amount of iron accumulated by Δyfh1 cells and improved their growth in an iron-dependent manner, and this effect was mostly transcriptional. Overexpressing the major mitochondrial phosphate carrier, MIR1, slightly increased the concentration of soluble mitochondrial phosphate and significantly improved various mitochondrial functions (cytochromes, [Fe-S] clusters, and respiration) in Δyfh1 cells. We conclude that in Δyfh1 cells, soluble phosphate is limiting, due to its co-precipitation with iron.

  19. A Nostoc punctiforme Sugar Transporter Necessary to Establish a Cyanobacterium-Plant Symbiosis1[C][W

    Science.gov (United States)

    Ekman, Martin; Picossi, Silvia; Campbell, Elsie L.; Meeks, John C.; Flores, Enrique

    2013-01-01

    In cyanobacteria-plant symbioses, the symbiotic nitrogen-fixing cyanobacterium has low photosynthetic activity and is supplemented by sugars provided by the plant partner. Which sugars and cyanobacterial sugar uptake mechanism(s) are involved in the symbiosis, however, is unknown. Mutants of the symbiotically competent, facultatively heterotrophic cyanobacterium Nostoc punctiforme were constructed bearing a neomycin resistance gene cassette replacing genes in a putative sugar transport gene cluster. Results of transport activity assays using 14C-labeled fructose and glucose and tests of heterotrophic growth with these sugars enabled the identification of an ATP-binding cassette-type transporter for fructose (Frt), a major facilitator permease for glucose (GlcP), and a porin needed for the optimal uptake of both fructose and glucose. Analysis of green fluorescent protein fluorescence in strains of N. punctiforme bearing frt::gfp fusions showed high expression in vegetative cells and akinetes, variable expression in hormogonia, and no expression in heterocysts. The symbiotic efficiency of N. punctiforme sugar transport mutants was investigated by testing their ability to infect a nonvascular plant partner, the hornwort Anthoceros punctatus. Strains that were specifically unable to transport glucose did not infect the plant. These results imply a role for GlcP in establishing symbiosis under the conditions used in this work. PMID:23463784

  20. Phosphate Tether-Mediated Ring-Closing Metathesis for the Generation of P-Stereogenic, Z-Configured Bicyclo[7.3.1]- and Bicyclo[8.3.1]phosphates.

    Science.gov (United States)

    Markley, Jana L; Maitra, Soma; Hanson, Paul R

    2016-02-05

    A phosphate tether-mediated ring-closing metathesis (RCM) study to the synthesis of Z-configured, P-stereogenic bicyclo[7.3.1]- and bicyclo[8.3.1]phosphates is reported. Investigations suggest that C3-substitution, olefin substitution, and proximity of the forming olefin to the bridgehead carbon of the bicyclic affect the efficiency and stereochemical outcome of the RCM event. This study demonstrates the utility of phosphate tether-mediated desymmetrization of C2-symmetric, 1,3-anti-diol-containing dienes in the generation of macrocyclic phosphates with potential synthetic and biological utility.

  1. Sphingosine-1-phosphate and renal vasoconstriction

    DEFF Research Database (Denmark)

    Jensen, Boye L

    2018-01-01

    ) and in conjunction with increased S1P release in pathophysiological situations like sepsis and ischemia-reperfusion incidents, this effect could be relevant in acute kidney injury with parallel decreases in renal blood flow and GFR. This article is protected by copyright. All rights reserved.......In the present issue of Acta Physiologica, Guan et al. in their article "Mechanisms of sphingosine-1-phosphate-mediated vasoconstriction of rat afferent arterioles" (1) address the signaling events associated with sphingosine-1-phosphate (S1P)-mediated renal afferent vasoconstriction and show in......, technically demanding, blood-perfused juxtamedullary nephron preparation that S1P signaling relies predominantly on transmembrane calcium influx from the extracellular fluid through L-type calcium channels with contribution from oxidative stress metabolites(1) . So not only is new information on S1P signaling...

  2. Chromatographic assay of degradation products of tributyl phosphate

    International Nuclear Information System (INIS)

    Tripathi, S.C.; Ramanujam, A.; Nadkarni, M.N.; Rao, K.A.; Bandyopadhyay, C.

    1985-01-01

    Gas-liquid chromatography (GLC) and thin-layer chromatographic (TLC) procedures have been developed for the estimation of monobutyl phosphoric acid (H 2 MBP) and dibutyl phosphoric acid (HDBP) present in tributyl phosphate (TBP). Less than one microgram of H 2 MBP and HDBP is visually detected on TLC plate made of silicagel-cellulose (1:1). HDBP as separated by TLC has been quantitatively estimated in the range of 40-260 μg using an indirect spectrophotometric method. GLC method using 10 percent (W/W) XE-60 column can be effectively used for the simultaneous determination of 0.25 percent W/V H 2 MBP and HDBP present in 30 percent TBP-n-dodecane. The sensitivity may be enhanced 100 fold by careful column conditioning and judicious control of operational parameters. (author)

  3. Oxidative stress as a mechanism of added sugar-induced cardiovascular disease.

    Science.gov (United States)

    Prasad, Kailash; Dhar, Indu

    2014-12-01

    Added sugars comprising of table sugar, brown sugar, corn syrup, maple syrup, honey, molasses, and other sweeteners in the prepared processed foods and beverages have been implicated in the pathophysiology of cardiovascular diseases. This article deals with the reactive oxygen species (ROS) as a mechanism of sugar-induced cardiovascular diseases. There is an association between the consumption of high levels of serum glucose with cardiovascular diseases. Various sources of sugar-induced generation of ROS, including mitochondria, nicotinamide adenine dinucleotide phosphate-oxidase, advanced glycation end products, insulin, and uric acid have been discussed. The mechanism by which ROS induce the development of atherosclerosis, hypertension, peripheral vascular disease, coronary artery disease, cardiomyopathy, heart failure, and cardiac arrhythmias have been discussed in detail. In conclusion, the data suggest that added sugars induce atherosclerosis, hypertension, peripheral vascular disease, coronary artery disease, cardiomyopathy, heart failure, and cardiac arrhythmias and that these effects of added sugars are mediated through ROS.

  4. Radiation chemistry of DNA. II. Strand breaks and sugar release by. gamma. -irradiation of DNA in aqueous solution. The effect of oxygen

    Energy Technology Data Exchange (ETDEWEB)

    Dizdaroglu, M; Schulte-Frohlinde, D; von Sonntag, C [Max-Planck-Institut fuer Kohlenforschung, Muelheim an der Ruhr (F.R. Germany). Abt. fuer Strahlenchemie

    1975-11-01

    From ..gamma..-irradiated oxygenated aqueous solutions of DNA, 2-deoxy-tetrodialdose (1) and 2-deoxy-pentos-4-ulose (2) have been isolated as free sugars. The formation of 1 indicates that in the presence of oxygen the DNA strand is not only broken by mere phosphate ester cleavage but also by C-C-bond rupture in the sugar moiety. Such a process has not been encountered in deoxygenated solutions so far. The mechanism for the formation of 1 is discussed.

  5. Molecular cloning, characterization and expression analysis of two members of the Pht1 family of phosphate transporters in Glycine max.

    Directory of Open Access Journals (Sweden)

    Zhaoyun Wu

    Full Text Available BACKGROUND: Phosphorus is one of the macronutrients essential for plant growth and development. The acquisition and translocation of phosphate are pivotal processes of plant growth. In a large number of plants, phosphate uptake by roots and translocation within the plant are presumed to occur via a phosphate/proton cotransport mechanism. PRINCIPAL FINDINGS: We cloned two cDNAs from soybean (Glycine max, GmPT1 and GmPT2, which show homology to the phosphate/proton cotransporter PHO84 from the budding yeast Saccharomyces cerevisiae. The amino acid sequence of the products predicted from GmPT1 and GmPT2 share 61% and 63% identity, respectively, with the PHO84 in amino acid sequence. The deduced structure of the encoded proteins revealed 12 membrane-spanning domains with a central hydrophilic region. The molecular mass values are ∼58.7 kDa for GmPT1 and ∼58.6 kDa for GmPT2. Transiently expressed GFP-protein fusions provide direct evidence that the two Pi transporters are located in the plasma membrane. Uptake of radioactive orthophosphate by the yeast mutant MB192 showed that GmPT1 and GmPT2 are dependent on pH and uptake is reduced by the addition of uncouplers of oxidative phosphorylation. The K(m for phosphate uptake by GmPT1 and GmPT2 is 6.65 mM and 6.63 mM, respectively. A quantitative real time RT-PCR assay indicated that these two genes are expressed in the roots and shoots of seedlings whether they are phosphate-deficient or not. Deficiency of phosphorus caused a slight change of the expression levels of GmPT1 and GmPT2. CONCLUSIONS: The results of our experiments show that the two phosphate transporters have low affinity and the corresponding genes are constitutively expressed. Thereby, the two phosphate transporters can perform translocation of phosphate within the plant.

  6. BRI1 and BAK1 interact with G proteins and regulate sugar-responsive growth and development in Arabidopsis.

    Science.gov (United States)

    Peng, Yuancheng; Chen, Liangliang; Li, Shengjun; Zhang, Yueying; Xu, Ran; Liu, Zupei; Liu, Wuxia; Kong, Jingjing; Huang, Xiahe; Wang, Yingchun; Cheng, Beijiu; Zheng, Leiying; Li, Yunhai

    2018-04-18

    Sugars function as signal molecules to regulate growth, development, and gene expression in plants, yeasts, and animals. A coordination of sugar availability with phytohormone signals is crucial for plant growth and development. The molecular link between sugar availability and hormone-dependent plant growth are largely unknown. Here we report that BRI1 and BAK1 are involved in sugar-responsive growth and development. Glucose influences the physical interactions and phosphorylations of BRI1 and BAK1 in a concentration-dependent manner. BRI1 and BAK1 physically interact with G proteins that are essential for mediating sugar signaling. Biochemical data show that BRI1 can phosphorylate G protein β subunit and γ subunits, and BAK1 can phosphorylate G protein γ subunits. Genetic analyses suggest that BRI1 and BAK1 function in a common pathway with G-protein subunits to regulate sugar responses. Thus, our findings reveal an important genetic and molecular mechanism by which BR receptors associate with G proteins to regulate sugar-responsive growth and development.

  7. The Effect of Phytase on the Oxygen Isotope Composition of Phosphate

    Science.gov (United States)

    von Sperber, C.; Tamburini, F.; Bernasconi, S. M.; Frossard, E.

    2013-12-01

    Plants and microorganisms under phosphorus (P) stress release extracellular phosphatases as a strategy to acquire inorganic phosphate (Pi) (1-2). These enzymes catalyze the hydrolysis of phosphoesters leading to a release of Pi. The enzymatic hydrolysis leads, via a nucleophilic attack, to the incorporation of one oxygen atom from the water into the newly formed Pi molecule. During the incorporation, an isotopic fractionation occurs, which might be used to identify the origin of Pi in the environment (3-6). While the effect of phosphomonoesterases and phosphodiesterases on the oxygen isotope composition of phosphate has been examined, there are, so far, no studies dealing with the effect of phytases (4-6). Phytases catalyze the hydrolysis of myo-inositol-hexakis-phosphate (IP6), which is an important component of organic P in many ecosystems (7). Enzymatic assays with phytase from wheat germ and Aspergillus niger were prepared under sterile and temperature controlled conditions in order to determine the effect of phytases on the oxygen isotope composition of phosphate, which has been liberated from IP6 via enzymatic hydrolysis. Assays with phytase from wheat germ lead to a turnover of the substrate close to 100%, while assays with phytase from Aspergillus niger lead to a turnover of the substrate close to 80%. In the case of the assays with phytase from wheat germ, our results indicate that one sixth of the total 24 oxygen which are associated to the phosphates in IP6 are exchanged with oxygen from water. From this we conclude that the incorporation of one oxygen atom from water occurs only at four phosphate molecules of IP6, while two phosphate molecules do not experience an incorporation of oxygen. This suggests that during the enzymatic hydrolysis, four P-O bonds and two C-O bonds are broken. Provided that, the isotopic fractionation can be calculated with an isotopic mass balance resulting in -8.4‰ (×3.6 SD). This is a value very similar to those reported

  8. A novel rare sugar inhibitor of murine herpes simplex keratitis.

    Science.gov (United States)

    Muniruzzaman, Syed; McIntosh, Megan; Hossain, Ahamed; Izumori, Ken; Bhattacharjee, Partha S

    2016-06-01

    To determine the therapeutic efficacy of a novel rare sugar, l-psicose, for the treatment of HSV-1 induced herpetic stromal keratitis (HSK) in a mouse eye model. One rare sugar l-psicose was assayed for HSV-1 inhibition of in vitro virus adsorption. The IC50 and IC90 values of l-psicose were determined using plaque reduction assay (PRA) in CV-1 cell. Female Balb/c mice were corneally infected with HSV-1, strain KOS-GFP; A topical eye drop treatment of l-psicose was started 24 h after infection and continued four times daily for ten consecutive days. The severity of HSK was monitored by slit lamp examination in a masked fashion and Infectious HSV-1 shedding was determined by PRA. l-psicose was found to have anti-viral activity in vitro at an IC50 dose of 99.5 mM and an IC90 dose of 160 mM. Topical eye drop treatment with 200 mM l-psicose in PBS solution significantly reduced the severity of HSK compared to the mock treatment group. The in vivo mouse ocular model results of l-psicose therapy correlated with accelerated clearance of virus from eye swabs. The results suggest that topical treatment with rare sugar l-psicose has efficacy against HSK through inhibition of HSV-1. Copyright © 2016 The Authors. Production and hosting by Elsevier B.V. All rights reserved.

  9. Total antioxidant content of alternatives to refined sugar.

    Science.gov (United States)

    Phillips, Katherine M; Carlsen, Monica H; Blomhoff, Rune

    2009-01-01

    Oxidative damage is implicated in the etiology of cancer, cardiovascular disease, and other degenerative disorders. Recent nutritional research has focused on the antioxidant potential of foods, while current dietary recommendations are to increase the intake of antioxidant-rich foods rather than supplement specific nutrients. Many alternatives to refined sugar are available, including raw cane sugar, plant saps/syrups (eg, maple syrup, agave nectar), molasses, honey, and fruit sugars (eg, date sugar). Unrefined sweeteners were hypothesized to contain higher levels of antioxidants, similar to the contrast between whole and refined grain products. To compare the total antioxidant content of natural sweeteners as alternatives to refined sugar. The ferric-reducing ability of plasma (FRAP) assay was used to estimate total antioxidant capacity. Major brands of 12 types of sweeteners as well as refined white sugar and corn syrup were sampled from retail outlets in the United States. Substantial differences in total antioxidant content of different sweeteners were found. Refined sugar, corn syrup, and agave nectar contained minimal antioxidant activity (sugar had a higher FRAP (0.1 mmol/100 g). Dark and blackstrap molasses had the highest FRAP (4.6 to 4.9 mmol/100 g), while maple syrup, brown sugar, and honey showed intermediate antioxidant capacity (0.2 to 0.7 mmol FRAP/100 g). Based on an average intake of 130 g/day refined sugars and the antioxidant activity measured in typical diets, substituting alternative sweeteners could increase antioxidant intake an average of 2.6 mmol/day, similar to the amount found in a serving of berries or nuts. Many readily available alternatives to refined sugar offer the potential benefit of antioxidant activity.

  10. Lysophosphatidic acid activates peroxisome proliferator activated receptor-γ in CHO cells that over-express glycerol 3-phosphate acyltransferase-1.

    Directory of Open Access Journals (Sweden)

    Cliona M Stapleton

    2011-04-01

    Full Text Available Lysophosphatidic acid (LPA is an agonist for peroxisome proliferator activated receptor-γ (PPARγ. Although glycerol-3-phosphate acyltransferase-1 (GPAT1 esterifies glycerol-3-phosphate to form LPA, an intermediate in the de novo synthesis of glycerolipids, it has been assumed that LPA synthesized by this route does not have a signaling role. The availability of Chinese Hamster Ovary (CHO cells that stably overexpress GPAT1, allowed us to analyze PPARγ activation in the presence of LPA produced as an intracellular intermediate. LPA levels in CHO-GPAT1 cells were 6-fold higher than in wild-type CHO cells, and the mRNA abundance of CD36, a PPARγ target, was 2-fold higher. Transactivation assays showed that PPARγ activity was higher in the cells that overexpressed GPAT1. PPARγ activity was enhanced further in CHO-GPAT1 cells treated with the PPARγ ligand troglitazone. Extracellular LPA, phosphatidic acid (PA or a membrane-permeable diacylglycerol had no effect, showing that PPARγ had been activated by LPA generated intracellularly. Transient transfection of a vector expressing 1-acylglycerol-3-phosphate acyltransferase-2, which converts endogenous LPA to PA, markedly reduced PPARγ activity, as did over-expressing diacylglycerol kinase, which converts DAG to PA, indicating that PA could be a potent inhibitor of PPARγ. These data suggest that LPA synthesized via the glycerol-3-phosphate pathway can activate PPARγ and that intermediates of de novo glycerolipid synthesis regulate gene expression.

  11. Sugar Allocation to Metabolic Pathways is Tightly Regulated and Affects the Virulence of Streptococcus mutans

    Directory of Open Access Journals (Sweden)

    Miki Kawada-Matsuo

    2016-12-01

    Full Text Available Bacteria take up and metabolize sugar as a carbohydrate source for survival. Most bacteria can utilize many sugars, including glucose, sucrose, and galactose, as well as amino sugars, such as glucosamine and N-acetylglucosamine. After entering the cytoplasm, the sugars are mainly allocated to the glycolysis pathway (energy production and to various bacterial component biosynthesis pathways, including the cell wall, nucleic acids and amino acids. Sugars are also utilized to produce several virulence factors, such as capsule and lipoteichoic acid. Glutamine-fructose-6-phosphate aminotransferase (GlmS and glucosamine-6-phosphate deaminase (NagB have crucial roles in sugar distribution to the glycolysis pathway and to cell wall biosynthesis. In Streptococcus mutans, a cariogenic pathogen, the expression levels of glmS and nagB are coordinately regulated in response to the presence or absence of amino sugars. In addition, the disruption of this regulation affects the virulence of S. mutans. The expression of nagB and glmS is regulated by NagR in S. mutans, but the precise mechanism underlying glmS regulation is not clear. In Staphylococcus aureus and Bacillus subtilis, the mRNA of glmS has ribozyme activity and undergoes self-degradation at the mRNA level. However, there is no ribozyme activity region on glmS mRNA in S. mutans. In this review article, we summarize the sugar distribution, particularly the coordinated regulation of GlmS and NagB expression, and its relationship with the virulence of S. mutans.

  12. Sphingosine-1-Phosphate (S1P)-Related Response of Human Conjunctival Fibroblasts After Filtration Surgery for Glaucoma.

    Science.gov (United States)

    Aoyama-Araki, Yuka; Honjo, Megumi; Uchida, Takatoshi; Yamagishi, Reiko; Kano, Kuniyuki; Aoki, Junken; Aihara, Makoto

    2017-04-01

    To investigate levels of sphingosine-1-phosphate (S1P) in aqueous fluid samples taken before and after filtration surgery and S1P-induced human conjunctival fibroblast (HCF) responses. Levels of S1P and its related sphingophospholipids in aqueous fluid obtained immediately before and after filtration surgery were determined by liquid chromatography-tandem mass spectrometry. HCFs were used for all in vitro experiments. The expression of five S1P receptor subtypes in HCFs was examined by quantitative real-time PCR. The effect of S1P and receptor-specific antagonists on HCF viability and cell migration was assessed by WST-1 assay and scratch migration assay, respectively. Differentiation to myofibroblasts and extracellular matrix production was evaluated by examining changes in F-actin, α-smooth muscle actin (αSMA), and collagen expression with immunocytochemistry, Western blotting, and collagen accumulation assay, respectively. No significant S1P levels in the aqueous fluid samples were detectable immediately before surgery, but postoperative levels of several lysophospholipids, including S1P, dehydro-S1P, and sphingosine, were significantly increased to bioactive concentrations in aqueous fluid in the blebs (P S1P receptor subtypes was detected in HCFs. Although S1P levels did not influence HCF proliferation, S1P enhanced cell migration, which could be inhibited by the S1P2 antagonist JTE 013. F-actin, αSMA, and collagen expression was significantly increased by S1P stimulation and was reduced by JTE 013. Bioactive S1P concentrations were present in the aqueous fluid at the end of filtration surgery. S1P activated HCFs via S1P2 receptors. These results revealed the potential of S1P2 antagonists in preventing scarring after glaucoma filtration surgery.

  13. High inorganic phosphate causes DNMT1 phosphorylation and subsequent fibrotic fibroblast activation

    Energy Technology Data Exchange (ETDEWEB)

    Tan, Xiaoying [Department of Nephrology and Rheumatology, Göttingen University Medical Center, Georg August University, Göttingen (Germany); Department of Cardiology and Pneumology, Göttingen University Medical Center, Georg August University, Göttingen (Germany); Xu, Xingbo [Department of Cardiology and Pneumology, Göttingen University Medical Center, Georg August University, Göttingen (Germany); Zeisberg, Elisabeth M. [Department of Cardiology and Pneumology, Göttingen University Medical Center, Georg August University, Göttingen (Germany); German Center for Cardiovascular Research (DZHK), Göttingen (Germany); Zeisberg, Michael, E-mail: mzeisberg@med.uni-goettingen.de [Department of Nephrology and Rheumatology, Göttingen University Medical Center, Georg August University, Göttingen (Germany); German Center for Cardiovascular Research (DZHK), Göttingen (Germany)

    2016-04-08

    Phosphate is an essential constituent of critical cellular functions including energy metabolism, nucleic acid synthesis and phosphorylation-dependent cell signaling. Increased plasma phosphate levels are an independent risk factor for lowered life-expectancy as well as for heart and kidney failure. Nevertheless, direct cellular effects of elevated phosphate concentrations within the microenvironment are poorly understood and have been largely neglected in favor of phosphor-regulatory hormones. Because interstitial fibrosis is the common determinant of chronic progressive kidney disease, and because fibroblasts are major mediators of fibrogenesis, we here explored the effect of high extracellular phosphate levels on renal fibroblasts. We demonstrate that high inorganic phosphate directly induces fibrotic fibroblast activation associated with increased proliferative activity, increased expression of α-smooth muscle actin and increased synthesis of type I collagen. We further demonstrate that such fibroblast activation is dependent on phosphate influx, aberrant phosphorylation of DNA methyltransferase DNMT1 and aberrant CpG island promoter methylation. In summary, our studies demonstrate that elevated phosphate concentrations induce pro-fibrotic fibroblast activation independent of phospho-regulatory hormones. - Highlights: • We exposed human kidney fibroblasts to media containing 1 mM or 3 mM phosphate. • Increased phosphate influx causes phosphorylation of DNA methyltransferase Dnmt1. • Phosphorylated Dnmt1 causes promoter methylation and transcriptional silencing of RASAL1. • Depletion of RASAL1 causes increased intrinsic Ras-GTP activity and fibroblast activation. • Inorganic phosphate causes fibroblast activation independent of phospho-regulatory hormones.

  14. Partial characterization of the cross-reacting determinant, a carbohydrate epitope shared by decay accelerating factor and the variant surface glycoprotein of the African Trypanosoma brucei.

    Science.gov (United States)

    Shak, S; Davitz, M A; Wolinsky, M L; Nussenzweig, V; Turner, M J; Gurnett, A

    1988-03-15

    The variant surface glycoprotein (VSG) of the African trypanosome is anchored in the cell membrane by a complex glycan attached to phosphatidylinositol. The carboxyl terminal portion of VSG contains a cryptic carbohydrate epitope, the cross-reacting determinant (CRD), that is revealed only after removal of the diacylglycerol by phosphatidylinositol-specific phospholipase C (PIPLC) or VSG lipase. Recently, we have shown that after hydrolysis by PIPLC, decay-accelerating factor (DAF)--a mammalian phosphatidylinositol-anchored protein--also contains the CRD epitope. Using a two site immunoradiometric assay in which the capturing antibody is a monoclonal antibody to DAF and the revealing antibody is anti-CRD, we now show that sugar phosphates significantly inhibited the binding of anti-CRD antibody to DAF released by PIPLC. DL-myo-inositol 1,2-cyclic phosphate was the most potent inhibitor of binding (IC50 less than 10(-8) M). Other sugar phosphates, such as alpha-D-glucose-1-phosphate, which also possess adjacent hydroxyl and phosphate moieties in cis also inhibited binding at low concentrations (IC50 = 10(-5) to 10(-4) M). In contrast, sugar phosphates which do not possess adjacent hydroxyl and phosphate moieties in cis and simple sugars weakly inhibited binding (IC50 greater than 10(-3) M). These results suggest that myo-inositol 1,2-cyclic phosphate contributes significantly to the epitope recognized by the anti-CRD antibody and is consistent with analysis of the carboxyl terminus of VSG, which also suggested the presence of the cyclic inositol phosphate. In light of the recent findings that human serum contains a glycan-phosphatidyl-inositol-specific phospholipase D, which converts DAF from a hydrophobic to a hydrophilic form lacking the CRD, the observation that the phosphate is crucial for expression of the epitope may be relevant in understanding the origin of CRD-negative DAF in urine and plasma.

  15. Elucidation of the regulatory role of the fructose operon reveals a novel target for enhancing the NADPH supply in Corynebacterium glutamicum

    DEFF Research Database (Denmark)

    Wang, Zhihao; Chan, Siu Hung Joshua; Sudarsan, Suresh

    2016-01-01

    is linked to redox and to the general metabolism. We here provide new insights into the regulation of the metabolism of this important platform organism by systematically characterizing mutants carrying various lesions in the fructose operon. Initially, we found that a strain where the dedicated fructose...... uptake system had been inactivated (KO-ptsF) was hampered in growth on sucrose minimal medium, and suppressor mutants appeared readily. Comparative genomic analysis in conjunction with enzymatic assays revealed that suppression was linked to inactivation of the pfkB gene, encoding a fructose-1-phosphate...... kinase. Detailed characterization of KO-ptsF, KO-pfkB and double knock-out (DKO) derivatives revealed a strong role for sugar-phosphates, especially fructose-1-phosphate (F1P), in governing sugar as well as redox metabolism due to effects on transcriptional regulation of key genes. These findings allowed...

  16. Phosphorylation and 14-3-3 binding of Arabidopsis trehalose-phosphate synthase 5 in response to 2-deoxyglucose

    DEFF Research Database (Denmark)

    Harthill, Jean E; Meek, Sarah E M; Morrice, Nick

    2006-01-01

    Trehalose-6-phosphate is a 'sugar signal' that regulates plant metabolism and development. The Arabidopsis genome encodes trehalose-6-phosphate synthase (TPS) and trehalose-6-phosphatase (TPP) enzymes. It also encodes class II proteins (TPS isoforms 5-11) that contain both TPS-like and TPP...

  17. The Role of Sphingosine-1-Phosphate and Ceramide-1-Phosphate in Inflammation and Cancer

    Directory of Open Access Journals (Sweden)

    Nitai C. Hait

    2017-01-01

    Full Text Available Inflammation is part of our body’s response to tissue injury and pathogens. It helps to recruit various immune cells to the site of inflammation and activates the production of mediators to mobilize systemic protective processes. However, chronic inflammation can increase the risk of diseases like cancer. Apart from cytokines and chemokines, lipid mediators, particularly sphingosine-1-phosphate (S1P and ceramide-1-phosphate (C1P, contribute to inflammation and cancer. S1P is an important player in inflammation-associated colon cancer progression. On the other hand, C1P has been recognized to be involved in cancer cell growth, migration, survival, and inflammation. However, whether C1P is involved in inflammation-associated cancer is not yet established. In contrast, few studies have also suggested that S1P and C1P are involved in anti-inflammatory pathways regulated in certain cell types. Ceramide is the substrate for ceramide kinase (CERK to yield C1P, and sphingosine is phosphorylated to S1P by sphingosine kinases (SphKs. Biological functions of sphingolipid metabolites have been studied extensively. Ceramide is associated with cell growth inhibition and enhancement of apoptosis while S1P and C1P are associated with enhancement of cell growth and survival. Altogether, S1P and C1P are important regulators of ceramide level and cell fate. This review focuses on S1P and C1P involvement in inflammation and cancer with emphasis on recent progress in the field.

  18. A phosphate-starvation-inducible outermembrane protein of Pseudomonas fluorescens Ag1 as an immunological phosphate-starvation marker

    DEFF Research Database (Denmark)

    Leopold, Kristine; Jacobsen, Susanne; Nybroe, Ole

    1997-01-01

    A phosphate-starvation-inducible outer-membrane protein of Pseudomonas fluorescens Ag1, expressed at phosphate concentrations below0.08-0.13 mM, was purified and characterized. The purification method involved separation of outer-membrane proteins by SDS-PAGE andextraction of the protein from...... nitrocellulose or PVDF membranes after electrotransfer of proteins to the membranes. The N-terminal amino acidsequence of the purified protein, called Psi1, did not show homology to any known proteins, and in contrast to the phosphate-specific porin OprP ofP. aeruginosa its mobility in SDS-PAGE was not affected...

  19. Gene Cloning of Iranian Leishmania major Mannose-1-Phosphate Guanyltransferase

    Directory of Open Access Journals (Sweden)

    R Salehi

    2009-07-01

    Full Text Available "nBackground: Leishmania is an obligatory intracellular protozoan parasite, which infects human be­ings when infected sand fly vector takes a blood meal.  Most efforts are towards designing an effective vaccine to prevent leishmaniasis. In this way, development of candidate antigen for vaccine has spe­cial im­portant. In this study, we cloned mannose-1-phosphate guanyltransferase gene of Iranian L .major in pET32a expression vector. "nMethods: Primers based on L. major mannose-1-phosphate guanyltransferase sequence gene was de­signed and synthesized. DNA of Leishmania promastigotes was extracted and PCR reaction was done. PCR product was cloned into pTZ57R and sub cloned into pET32a expression vector. "nResults: Recombinant plasmid containing 1140 bp as L. major mannose-1-phosphate guanyltrans­ferase gene was extracted and confirmed by restriction analysis. PCR product was sequenced and de­posited to GenBank. There were some differences in amino acid sequences between Iranian L. major mannose-1-phosphate guanyltransferase and others previously accepted in GenBank "nConclusion: We amplified and cloned Iranian L. major mannose-1-phosphate guanyltransferase successfully.

  20. A European multicientre study on the comparison of HIV-1 viral loads between VERIS HIV-1 Assay and Roche COBAS® TAQMAN® HIV-1 test, Abbott RealTime HIV-1 Assay, and Siemens VERSANT HIV-1 Assay.

    Science.gov (United States)

    Braun, Patrick; Delgado, Rafael; Drago, Monica; Fanti, Diana; Fleury, Hervé; Hofmann, Jörg; Izopet, Jacques; Kühn, Sebastian; Lombardi, Alessandra; Mancon, Alessandro; Marcos, Mª Angeles; Mileto, Davide; Sauné, Karine; O'Shea, Siobhan; Pérez-Rivilla, Alfredo; Ramble, John; Trimoulet, Pascale; Vila, Jordi; Whittaker, Duncan; Artus, Alain; Rhodes, Daniel

    2017-07-01

    Viral load monitoring is essential for patients under treatment for HIV. Beckman Coulter has developed the VERIS HIV-1 Assay for use on the novel, automated DxN VERIS Molecular Diagnostics System. ¥ OBJECTIVES: Evaluation of the clinical performance of the new quantitative VERIS HIV-1 Assay at multiple EU laboratories. Method comparison with the VERIS HIV-1 Assay was performed with 415 specimens at 5 sites tested with COBAS ® AmpliPrep/COBAS ® TaqMan ® HIV-1 Test, v2.0, 169 specimens at 3 sites tested with RealTime HIV-1 Assay, and 202 specimens from 2 sites tested with VERSANT HIV-1 Assay. Patient monitoring sample results from 4 sites were also compared. Bland-Altman analysis showed the average bias between VERIS HIV-1 Assay and COBAS HIV-1 Test, RealTime HIV-1 Assay, and VERSANT HIV-1 Assay to be 0.28, 0.39, and 0.61 log 10 cp/mL, respectively. Bias at low end levels below 1000cp/mL showed predicted bias to be <0.3 log 10 cp/mL for VERIS HIV-1 Assay versus COBAS HIV-1 Test and RealTime HIV-1 Assay, and <0.5 log 10 cp/mL versus VERSANT HIV-1 Assay. Analysis on 174 specimens tested with the 0.175mL volume VERIS HIV-1 Assay and COBAS HIV-1 Test showed average bias of 0.39 log 10 cp/mL. Patient monitoring results using VERIS HIV-1 Assay demonstrated similar viral load trends over time to all comparators. The VERIS HIV-1 Assay for use on the DxN VERIS System demonstrated comparable clinical performance to COBAS ® HIV-1 Test, RealTime HIV-1 Assay, and VERSANT HIV-1 Assay. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. The return of metabolism: biochemistry and physiology of the pentose phosphate pathway

    Science.gov (United States)

    Stincone, Anna; Prigione, Alessandro; Cramer, Thorsten; Wamelink, Mirjam M. C.; Campbell, Kate; Cheung, Eric; Olin-Sandoval, Viridiana; Grüning, Nana-Maria; Krüger, Antje; Alam, Mohammad Tauqeer; Keller, Markus A.; Breitenbach, Michael; Brindle, Kevin M.; Rabinowitz, Joshua D.; Ralser, Markus

    2015-01-01

    The pentose phosphate pathway (PPP) is a fundamental component of cellular metabolism. The PPP is important to maintain carbon homoeostasis, to provide precursors for nucleotide and amino acid biosynthesis, to provide reducing molecules for anabolism, and to defeat oxidative stress. The PPP shares reactions with the Entner–Doudoroff pathway and Calvin cycle and divides into an oxidative and non-oxidative branch. The oxidative branch is highly active in most eukaryotes and converts glucose 6-phosphate into carbon dioxide, ribulose 5-phosphate and NADPH. The latter function is critical to maintain redox balance under stress situations, when cells proliferate rapidly, in ageing, and for the ‘Warburg effect’ of cancer cells. The non-oxidative branch instead is virtually ubiquitous, and metabolizes the glycolytic intermediates fructose 6-phosphate and glyceraldehyde 3-phosphate as well as sedoheptulose sugars, yielding ribose 5-phosphate for the synthesis of nucleic acids and sugar phosphate precursors for the synthesis of amino acids. Whereas the oxidative PPP is considered unidirectional, the non-oxidative branch can supply glycolysis with intermediates derived from ribose 5-phosphate and vice versa, depending on the biochemical demand. These functions require dynamic regulation of the PPP pathway that is achieved through hierarchical interactions between transcriptome, proteome and metabolome. Consequently, the biochemistry and regulation of this pathway, while still unresolved in many cases, are archetypal for the dynamics of the metabolic network of the cell. In this comprehensive article we review seminal work that led to the discovery and description of the pathway that date back now for 80 years, and address recent results about genetic and metabolic mechanisms that regulate its activity. These biochemical principles are discussed in the context of PPP deficiencies causing metabolic disease and the role of this pathway in biotechnology, bacterial and

  2. 26 CFR 301.6511(e)-1 - Special rules applicable to manufactured sugar.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 18 2010-04-01 2010-04-01 false Special rules applicable to manufactured sugar... Assessment and Collection § 301.6511(e)-1 Special rules applicable to manufactured sugar. (a) Use as... the person entitled thereto. Such right accrues as of the date the manufactured sugar, or article...

  3. Saccharification of recalcitrant biomass and integration options for lignocellulosic sugars from Catchlight Energy's sugar process (CLE Sugar).

    Science.gov (United States)

    Gao, Johnway; Anderson, Dwight; Levie, Benjamin

    2013-01-28

    Woody biomass is one of the most abundant biomass feedstocks, besides agriculture residuals in the United States. The sustainable harvest residuals and thinnings alone are estimated at about 75 million tons/year. These forest residuals and thinnings could produce the equivalent of 5 billion gallons of lignocellulosic ethanol annually. Softwood biomass is the most recalcitrant biomass in pretreatment before an enzymatic hydrolysis. To utilize the most recalcitrant lignocellulosic materials, an efficient, industrially scalable and cost effective pretreatment method is needed. Obtaining a high yield of sugar from recalcitrant biomass generally requires a high severity of pretreatment with aggressive chemistry, followed by extensive conditioning, and large doses of enzymes. Catchlight Energy's Sugar process, CLE Sugar, uses a low intensity, high throughput variation of bisulfite pulping to pretreat recalcitrant biomass, such as softwood forest residuals. By leveraging well-proven bisulfite technology and the rapid progress of enzyme suppliers, CLE Sugar can achieve a high yield of total biomass carbohydrate conversion to monomeric lignocellulosic sugars. For example, 85.8% of biomass carbohydrates are saccharified for un-debarked Loblolly pine chips (softwood), and 94.0% for debarked maple chips (hardwood). Furan compound formation was 1.29% of biomass feedstock for Loblolly pine and 1.10% for maple. At 17% solids hydrolysis of pretreated softwood, an enzyme dose of 0.075 g Sigma enzyme mixture/g dry pretreated (unwashed) biomass was needed to achieve 8.1% total sugar titer in the hydrolysate and an overall prehydrolysate liquor plus enzymatic hydrolysis conversion yield of 76.6%. At a much lower enzyme dosage of 0.044 g CTec2 enzyme product/g dry (unwashed) pretreated softwood, hydrolysis at 17% solids achieved 9.2% total sugar titer in the hydrolysate with an overall sugar yield of 85.0% in the combined prehydrolysate liquor and enzymatic hydrolysate. CLE Sugar has

  4. Therapeutic targeting of Neu1 sialidase with oseltamivir phosphate (Tamiflu® disables cancer cell survival in human pancreatic cancer with acquired chemoresistance

    Directory of Open Access Journals (Sweden)

    O’Shea LK

    2014-01-01

    Full Text Available Leah K O'Shea,1 Samar Abdulkhalek,1 Stephanie Allison,2 Ronald J Neufeld,2 Myron R Szewczuk11Department of Biomedical and Molecular Sciences, 2Department of Chemical Engineering, Queen's University, Kingston, ON, CanadaBackground: Resistance to drug therapy, along with high rates of metastasis, contributes to the low survival rate in patients diagnosed with pancreatic cancer. An alternate treatment for human pancreatic cancer involving targeting of Neu1 sialidase with oseltamivir phosphate (Tamiflu® was investigated in human pancreatic cancer (PANC1 cells with acquired resistance to cisplatin and gemcitabine. Its efficacy in overcoming the intrinsic resistance of the cell to chemotherapeutics and metastasis was evaluated.Methods: Microscopic imaging, immunocytochemistry, immunohistochemistry, and WST-1 cell viability assays were used to evaluate cell survival, morphologic changes, and expression levels of E-cadherin, N-cadherin, and VE-cadherin before and after treatment with oseltamivir phosphate in PANC1 cells with established resistance to cisplatin, gemcitabine, or a combination of the two agents, and in archived paraffin-embedded PANC1 tumors grown in RAGxCγ double mutant mice.Results: Oseltamivir phosphate overcame the chemoresistance of PANC1 to cisplatin and gemcitabine alone or in combination in a dose-dependent manner, and disabled the cancer cell survival mechanism(s. Oseltamivir phosphate also reversed the epithelial-mesenchymal transition characteristic of the phenotypic E-cadherin to N-cadherin changes associated with resistance to drug therapy. Low-dose oseltamivir phosphate alone or in combination with gemcitabine in heterotopic xenografts of PANC1 tumors growing in RAGxCγ double mutant mice did not prevent metastatic spread to the liver and lung.Conclusion: Therapeutic targeting of Neu1 sialidase with oseltamivir phosphate at the growth factor receptor level disables the intrinsic signaling platform for cancer cell survival

  5. Effects of sugar solutions on hypothalamic appetite regulation.

    Science.gov (United States)

    Colley, Danielle L; Castonguay, Thomas W

    2015-02-01

    Several hypotheses for the causes of the obesity epidemic in the US have been proposed. One such hypothesis is that dietary intake patterns have significantly shifted to include unprecedented amounts of refined sugar. We set out to determine if different sugars might promote changes in the hypothalamic mechanisms controlling food intake by measuring several hypothalamic peptides subsequent to overnight access to dilute glucose, sucrose, high fructose corn syrup, or fructose solutions. Rats were given access to food, water and a sugar solution for 24h, after which blood and tissues were collected. Fructose access (as opposed to other sugars that were tested) resulted in a doubling of circulating triglycerides. Glucose consumption resulted in upregulation of 7 satiety-related hypothalamic peptides whereas changes in gene expression were mixed for remaining sugars. Also, following multiple verification assays, 6 satiety related peptides were verified as being affected by sugar intake. These data provide evidence that not all sugars are equally effective in affecting the control of intake. Copyright © 2014. Published by Elsevier Inc.

  6. Source/ sink interactions underpin crop yield: the case for trehalose 6-phosphate/ SnRK1 in improvement of wheat

    Directory of Open Access Journals (Sweden)

    Matthew ePaul

    2014-08-01

    Full Text Available Considerable interest has been evoked by the analysis of the regulatory pathway in carbohydrate metabolism and cell growth involving trehalose (TRE. TRE is at small concentrations in mesophytes such as Triticum aestivum. Studies of TRE metabolism, and genetic modification of it, have shown a very wide and important role of the pathway in regulation of many processes in development, growth and photosynthesis. It has now been established that trehalose 6-phosphate (T6P, is formed from glucose-6-phosphate and UDP-glucose, derived from sucrose, by the action of trehalose phosphate synthase (TPS and broken down by trehalose-6-phosphate phosphatase (TPP providing for subtle regulation. The concentration of T6P increases with sucrose concentration. Many of the effects of T6P on metabolism and growth occur via the interaction of T6P with the SnRK1 protein kinase system. A large concentration of sucrose increases T6P and thereby inhibits SnRK1, so stimulating growth of cells and their metabolism. The T6P/SnRK1 mechanism offers an important new view of how the distribution of assimilates to organs, such as developing cereal grains, is achieved. Changing T6P concentrations by genetically modifying TPS and TPP has altered photosynthesis, sugar metabolism, growth and development which affect responses to, and recovery from, environmental factors. This review briefly summarizes the factors determining, and limiting, yield of wheat, particularly mass/grain which is highly conserved. The interactions between the source and sink relations are addressed together with how T6P/SnRK1 might function to determine grain number, size, and yield. The possibility of how these might be increased by modifying trehalose metabolism is considered. Cereal yields globally are not increasing and careful targeting of T6P may offer a way of optimizing grain growth and thus increasing yield in wheat.

  7. Distribution of 226Ra, 232Th and 40K in soils and sugar cane crops at Corumbatai river basin, Sao Paulo State, Brazil

    International Nuclear Information System (INIS)

    Tomazini da Conceicao, Fabiano; Bonotto, Daniel Marcos; Jimenez-Rueda, Jairo Roberto; Frutuoso Roveda, Jose Arnaldo

    2009-01-01

    The common use of phosphate fertilizers NPK and amendments in sugar cane crops in Brazilian agriculture may increase the 226 Ra, 232 Th and 40 K activity concentrations in soils and their availability for plants and human food chain. Thus, the main aim of this study was to evaluate the distribution of 226 Ra, 232 Th and 40 K in soils and sugar cane crops in the Corumbatai river basin, Sao Paulo State, Brazil. The gamma spectrometry was utilized to measure the 226 Ra, 232 Th and 40 K activity concentration in all samples. The soil-to-sugar cane transfer factors (TF) were quantified using the ratio between the radionuclide activity concentration in sugar cane and its activity concentration in soil. The results show that, although radionuclides incorporated in phosphate fertilizers and amendments are annually added in the sugar cane crops, if utilized in accordance with the recommended rates, their use does not lead to hazards levels in soils. The soil-to-sugar cane transfer of radionuclides occurred in the following order 40 K> 226 Ra> 232 Th. Therefore, under these conditions, radionuclides intake through consumption of sugar is not hazardous to human health.

  8. Oxidized LDL-induced angiogenesis involves sphingosine 1-phosphate: prevention by anti-S1P antibody.

    Science.gov (United States)

    Camaré, Caroline; Trayssac, Magali; Garmy-Susini, Barbara; Mucher, Elodie; Sabbadini, Roger; Salvayre, Robert; Negre-Salvayre, Anne

    2015-01-01

    Neovascularization occurring in atherosclerotic lesions may promote plaque expansion, intraplaque haemorrhage and rupture. Oxidized LDL (oxLDL) are atherogenic, but their angiogenic effect is controversial; both angiogenic and anti-angiogenic effects have been reported. The angiogenic mechanism of oxLDL is partly understood, but the role of the angiogenic sphingolipid, sphingosine 1-phosphate (S1P), in this process is not known. Thus, we investigated whether S1P is involved in the oxLDL-induced angiogenesis and whether an anti-S1P monoclonal antibody can prevent this effect. Angiogenesis was assessed by capillary tube formation by human microvascular endothelial cells (HMEC-1) cultured on Matrigel and in vivo by the Matrigel plug assay in C57BL/6 mice. Human oxLDL exhibited a biphasic angiogenic effect on HMEC-1; low concentrations were angiogenic, higher concentrations were cytotoxic. The angiogenic response to oxLDL was blocked by the sphingosine kinase (SPHK) inhibitor, dimethylsphingosine, by SPHK1-siRNA and by an anti-S1P monoclonal antibody. Moreover, inhibition of oxLDL uptake and subsequent redox signalling by anti-CD36 and anti-LOX-1 receptor antibodies and by N-acetylcysteine, respectively, blocked SPHK1 activation and tube formation. In vivo, in the Matrigel plug assay, low concentrations of human oxLDL or murine oxVLDL also triggered angiogenesis, which was prevented by i.p. injection of the anti-S1P antibody. These data highlight the role of S1P in angiogenesis induced by oxLDL both in HMEC-1 cultured on Matrigel and in vivo in the Matrigel plug model in mice, and demonstrate that the anti-S1P antibody effectively blocks the angiogenic effect of oxLDL. © 2014 The British Pharmacological Society.

  9. Rice calcium-dependent protein kinase OsCPK17 targets plasma membrane intrinsic protein and sucrose phosphate synthase and is required for a proper cold stress response

    KAUST Repository

    Almadanim, M. Cecília

    2017-01-19

    Calcium-dependent protein kinases (CDPKs) are involved in plant tolerance mechanisms to abiotic stresses. Although CDPKs are recognized as key messengers in signal transduction, the specific role of most members of this family remains unknown. Here we test the hypothesis that OsCPK17 plays a role in rice cold stress response by analyzing OsCPK17 knockout, silencing, and overexpressing rice lines under low temperature. Altered OsCPK17 gene expression compromises cold tolerance performance, without affecting the expression of key cold stress-inducible genes. A comparative phosphoproteomic approach led to the identification of six potential in vivo OsCPK17 targets, which are associated with sugar and nitrogen metabolism, and with osmotic regulation. To test direct interaction, in vitro kinase assays were performed, showing that the sucrose phosphate synthase OsSPS4, and the aquaporin OsPIP2;1/OsPIP2;6 are phosphorylated by OsCPK17 in a calcium-dependent manner. Altogether, our data indicates that OsCPK17 is required for a proper cold stress response in rice, likely affecting the activity of membrane channels and sugar metabolism.

  10. Synthesis and Physicochemical Characterization of D-Tagatose-1-phosphate: The Substrate of the Tagatose-1-Phosphate Kinase TagK in the PTS-mediated D-Tagatose Catabolic Pathway of Bacillus licheniformis

    Science.gov (United States)

    Van der Heiden, Edwige; Delmarcelle, Michaël; Simon, Patricia; Counson, Melody; Galleni, Moreno; Freedberg, Darón I.; Thompson, John; Joris, Bernard; Battistel, Marcos D.

    2015-01-01

    We report the first enzymatic synthesis of D-tagatose-1-phosphate (Tag-1P) by the multi-component PEP-dependent:tag-PTS present in tagatose-grown cells of Klebsiella pneumoniae. Physicochemical characterization by 31P and 1H NMR spectroscopy reveals that, in solution, this derivative is primarily in the pyranose form. Tag-1P was used to characterize the putative tagatose-1-phosphate kinase (TagK) of the Bacillus licheniformis PTS-mediated D-Tagatose catabolic Pathway (Bli-TagP). For this purpose, a soluble protein fusion was obtained with the 6 His-tagged trigger factor (TFHis6) of Escherichia coli. The active fusion enzyme was named TagK-TFHis6. Tag-1P and D-fructose-1-phosphate (Fru-1P) are substrates for the TagK-TFHis6 enzyme, whereas the isomeric derivatives D-tagatose-6-phosphate (Tag-6P) and D-fructose-6-phosphate (Fru-6P) are inhibitors. Studies of catalytic efficiency (kcat/Km) reveal that the enzyme specificity is markedly in favor of Tag-1P as substrate. Importantly, we show in vivo that the transfer of the phosphate moiety from PEP to the B. licheniformis tagatose-specific enzyme II (EIITag) in E.coli is inefficient. The capability of the PTS general cytoplasmic components of B. subtilis, HPr and EI, to restore the phosphate transfer is demonstrated. PMID:26159072

  11. Continuous ethanol production using yeast immobilized on sugar-cane stalks

    Energy Technology Data Exchange (ETDEWEB)

    Vasconcelos, J.N. de [Alagoas Univ., Maceio, AL (Brazil). Dept. de Engenharia Quimica]. E-mail: jnunes@ctec.ufal.br; Lopes, C.E. [Pernambuco Univ., Recife, PE (Brazil). Dept. de Antibioticos; Franca, F.P. de [Universidade Federal, Rio de Janeiro, RJ (Brazil). Escola de Quimica. Dept. de Engenharia Bioquimica

    2004-09-01

    Sugar-cane stalks, 2.0 cm long, were used as a support for yeast immobilization envisaging ethanol production. The assays were conducted in 38.5 L fermenters containing a bed of stalks with 50% porosity. The operational stability of the immobilized yeast, the efficiency and stability of the process, as well as the best dilution rate were evaluated. Molasses from demerara sugar production was used in the medium formulation. It was diluted to obtain 111.75 {+-} 1.51 g/L without any further treatment. Sulfuric acid was used to adjust the pH value to around 4.2. Every two days Kamoran HJ (10 ppm) or with a mixture containing penicillin (10 ppm) and tetracycline (10 ppm), was added to the medium. Ethanol yield and efficiency were 29.64 g/L.h and 86.40%, respectively, and the total reducing sugars conversion was 74.61% at a dilution rate of 0.83 h{sup -1}. The yeast-stalk system was shown to be stable for over a 60 day period at extremely variable dilution rates ranging from 0.05 h{sup -1} to 3.00 h{sup -1}. The concentration of immobilized cell reached around 109 cells/gram of dry sugar-cane stalk when the fermenter was operating at the highest dilution rate (3.00 h{sup -1}). (author)

  12. Evaluation of the availability of phosphorus from decalcium phosphate and rock phosphates from Patos de Minas, Tapira and Finos de Tapira for sheep, by the isotope dilution technique

    International Nuclear Information System (INIS)

    Vitti, D.M.S.S.

    1989-01-01

    'In vitro' and 'in vivo' assays were carried out to determine the phosphorus availability from dicalcium phosphate and rock phosphates from Patos de Minas, Tapira and Finos de Tapira. Twenty four male sheep, with 40 kg live weight, were assigned to three groups of eight animals each. The animals were housed individually in metabolism cages and received a diet containing cassava meal, urea, molasses, soybean meal and mineral mixture. Phosphate sources were added to give 4 g of phosphorus per animal per day. After two weeks on the experimental diet each sheep was injected intravenously with 200 μCi of 32 P (Na 2 HPO 4 ). Blood samples were collected from the jugular vein at 24 hs intervals for 8 days. The daily fecal outputs were collected for 8 days and sampled. The specific activities of plasma and feces were determined and the fecal endogenous loss and true phosphorus absorption were calculated. For 'in vitro' assay, rumen samples were collected from a fistulated steer and aliquots were incubated with 0.1 μCi 32 P (Na 2 HPO 4 ) in a medium containing the phosphorus sources. After centrifugation microorganisms were separated and phosphorus incorporation determined. The true absorption values were 58.92; 50.85; 47.99 and 42.72% for dicalcium phosphate, Finos de Tapira, Tapira and Patos, respectively. Dicalcium phosphate showed higher availability (P [pt

  13. Fluorescence-based rapid measurement of sphingosine-1-phosphate transport activity in erythrocytes[S

    Science.gov (United States)

    Kobayashi, Naoki; Otsuka, Masato; Yamaguchi, Akihito; Nishi, Tsuyoshi

    2016-01-01

    Sphingosine-1-phosphate (S1P) is present in the blood plasma and acts as a pivotal intercellular signal transmitter in the immune system by recruiting lymphocytes from the thymus and secondary lymphoid tissues. The plasma S1P concentration is maintained by the supply of S1P from erythrocytes. Previously, we showed that S1P release from erythrocytes is mediated by an ATP-dependent transporter. In this study, we attempted to establish a rapid and reliable method for measuring the S1P transport activity in erythrocytes by using a fluorescent S1P analog, 7-nitro-2-1,3-benzoxadiazol-4-yl (NBD)-labeled S1P. NBD-S1P was released from erythrocytes in a time-dependent manner. The NBD-S1P release was reduced after exposure to glyburide, which is an inhibitor of the S1P transporter in erythrocytes. Moreover, the release of NBD-S1P and S1P from erythrocytes was competitively inhibited by intracellular S1P and NBD-S1P, respectively. These results showed that the erythrocyte S1P transporter exports NBD-S1P. We optimized the sample-preparation conditions and lipid extraction to increase the sensitivity of the assay. Furthermore, we successfully measured NBD-S1P release without lipid extraction by decreasing the concentration of BSA in the assay buffer to 0.1%. This method will be useful for the high-throughput screening of S1P transporter inhibitors using conventional fluorometers. PMID:27655910

  14. L-Myo-inositol 1-phosphate synthase in the aquatic fern Azolla filiculoides.

    Science.gov (United States)

    Benaroya, Rony Oren; Zamski, Eli; Tel-Or, Elisha

    2004-02-01

    L-Myo-inositol 1-phosphate synthase (INPS EC 5.5.1.4) catalyzes the conversion of D-glucose 6-phosphate to L-myo-inositol 1-phosphate. INPS is a key enzyme involved in the biosynthesis of phytate which is a common form of stored phosphates in higher plants. The present study monitored the increase of INPS expression in Azolla filiculoides resulting from exposure to inorganic phosphates, metals and salt stress. The expression of INPS was significantly higher in Azolla plants that were grown in rich mineral growth medium than those maintained on nutritional growth medium. The expression of INPS protein and corresponding mRNA increased in plants cultured in minimal nutritional growth medium when phosphate or Zn2+, Cd2+ and NaCl were added to the growth medium. When employing rich mineral growth medium, INPS protein content increased with the addition of Zn2+, but decreased in the presence of Cd2+ and NaCl. These results indicated that accumulation of phytate in Azolla is a result of the intensified expression of INPS protein and mRNA, and its regulation may be primarily derived by the uptake of inorganic phosphate, and Zn2+, Cd2+ or NaCl.

  15. Sugar recognition by human galactokinase

    Directory of Open Access Journals (Sweden)

    Timson David J

    2003-11-01

    Full Text Available Abstract Background Galactokinase catalyses the first committed step of galactose catabolism in which the sugar is phosphorylated at the expense of MgATP. Recent structural studies suggest that the enzyme makes several contacts with galactose – five side chain and two main chain hydrogen bonds. Furthermore, it has been suggested that inhibition of galactokinase may help sufferers of the genetic disease classical galactosemia which is caused by defects in another enzyme of the pathway galactose-1-phosphate uridyl transferase. Galactokinases from different sources have a range of substrate specificities and a diversity of kinetic mechanisms. Therefore only studies on the human enzyme are likely to be of value in the design of therapeutically useful inhibitors. Results Using recombinant human galactokinase expressed in and purified from E. coli we have investigated the sugar specificity of the enzyme and the kinetic consequences of mutating residues in the sugar-binding site in order to improve our understanding of substrate recognition by this enzyme. D-galactose and 2-deoxy-D-galactose are substrates for the enzyme, but N-acetyl-D-galactosamine, L-arabinose, D-fucose and D-glucose are all not phosphorylated. Mutation of glutamate-43 (which forms a hydrogen bond to the hydroxyl group attached to carbon 6 of galactose to alanine results in only minor changes in the kinetic parameters of the enzyme. Mutation of this residue to glycine causes a ten-fold drop in the turnover number. In contrast, mutation of histidine 44 to either alanine or isoleucine results in insoluble protein following expression in E. coli. Alteration of the residue that makes hydrogen bonds to the hydroxyl attached to carbons 3 and 4 (aspartate 46 results in an enzyme that although soluble is essentially inactive. Conclusions The enzyme is tolerant to small changes at position 2 of the sugar ring, but not at positions 4 and 6. The results from site directed mutagenesis could

  16. Platinum Catalyzed Ring-Opening of 1,2-Cyclopropanated Sugars with O-Nucleophiles

    DEFF Research Database (Denmark)

    Beyer, Jürgen; Skaanderup, Philip Robert; Madsen, Robert

    1999-01-01

    In the presence of a catalytic amount of Zeise's dimer 1,2-cyclopropanated sugars undergo regioselective ring-opening at C-1 with O-nucleophiles including alcohols, phenols and water to produce 2-C-branched carbohydrates.......In the presence of a catalytic amount of Zeise's dimer 1,2-cyclopropanated sugars undergo regioselective ring-opening at C-1 with O-nucleophiles including alcohols, phenols and water to produce 2-C-branched carbohydrates....

  17. Impacts of Sugar Import Policy on Sugar Production in Indonesia

    OpenAIRE

    Suryantoro, Agustinus; Susilo, Albertus Magnus; Supriyono, Supriyono

    2013-01-01

    Production of sugar unful lled consumption of Indonesia society. The lack of consumption and productionhave ful lled by import. Assumption national consumption 2,7 million ton, Indonesia will import sugar in 2013predicted about 300.000 ton (Tempo.co, August, 21, 2012).The aims in general of this research are to understand the impact of sugar import policy on sugar production.Especially (1) to understand the factors that in uence sugar import price, (2) to understand impact of sugarimport pric...

  18. Sugars, the clock and transition to flowering

    Directory of Open Access Journals (Sweden)

    Mohammad Reza eBolouri Moghaddam

    2013-02-01

    Full Text Available Sugars do not only act as source of energy, but they also act as signals in plants. This mini review summarizes the emerging links between sucrose-mediated signaling and the cellular networks involved in flowering time control and defense. Cross-talks with gibberellin (GA and jasmonate (JA signaling pathways are highlighted. The circadian clock fulfills a crucial role at the heart of cellular networks and the bilateral relation between sugar signaling and the clock is discussed. It is proposed that important factors controlling plant growth (DELLAs, PIFs, invertases and trehalose- 6-phosphate or T6P might fulfill central roles in the transition to flowering as well. The emerging concept of ‘sweet immunity’, modulated by the clock, might at least partly rely on a sucrose-specific signaling pathway that needs further exploration.

  19. Sap-Sugar Content of Grafted Sugar Maple Trees

    Science.gov (United States)

    Maurice E. Jr. Demeritt; Maurice E. Jr. Demeritt

    1985-01-01

    In March and April 1983, 289 and 196 young grafted sugar maple trees were tapped and evaluated for sap-sugar content. In April, sap was collected from taps both above and below the graft union. Diameter of all tapped trees at 18 inches above the ground was measured. Analysis of the data revealed that: (1) trees selected for high sugar yield cannot be reproduced by...

  20. Novel Platinum Catalyzed Glycosylation Using 1,2-Cyclopropanated Sugars as Glycosyl Donors

    DEFF Research Database (Denmark)

    Beyer, Jürgen; Madsen, Robert

    1998-01-01

    By treating 1,2-cyclopropanated sugars with Zeise's dimer and an alcohol a regioselective ring opening of the cyclopropane takes place to give 2-C-alkyl-2-deoxy-glycosides......By treating 1,2-cyclopropanated sugars with Zeise's dimer and an alcohol a regioselective ring opening of the cyclopropane takes place to give 2-C-alkyl-2-deoxy-glycosides...

  1. Real-time pcr (qpcr) assay for rhizoctonia solani anastomoses group ag2-2 iiib

    International Nuclear Information System (INIS)

    Abbas, S.J.; Ahmad, B.

    2014-01-01

    Rhizoctonia solani anastomosis group AG2-2 IIIB is a severe sugar beet and maize pathogen. It causes crown and root rot disease which leads to yield losses world-wide. The soil-borne pathogen is difficult to detect and quantify by conventional methods. We developed a real-time PCR (qPCR) assay for the quantification of genomic DNA of Rhizoctonia solani AG2-2 IIIB based on the ITS region of rDNA genes. The limit of quantification of the assay is 1.8 pg genomic DNA. The amplification efficiency was 96.4. The assay will be helpful in the diagnoses of Rhizoctonia solani infection of sugar beet and maize roots and in the quantification of R. solani AG2-2 IIIB inoculum in plant debris and soil. (author)

  2. Enzyme-linked electrochemical DNA ligation assay using magnetic beads.

    Science.gov (United States)

    Stejskalová, Eva; Horáková, Petra; Vacek, Jan; Bowater, Richard P; Fojta, Miroslav

    2014-07-01

    DNA ligases are essential enzymes in all cells and have been proposed as targets for novel antibiotics. Efficient DNA ligase activity assays are thus required for applications in biomedical research. Here we present an enzyme-linked electrochemical assay based on two terminally tagged probes forming a nicked junction upon hybridization with a template DNA. Nicked DNA bearing a 5' biotin tag is immobilized on the surface of streptavidin-coated magnetic beads, and ligated product is detected via a 3' digoxigenin tag recognized by monoclonal antibody-alkaline phosphatase conjugate. Enzymatic conversion of napht-1-yl phosphate to napht-1-ol enables sensitive detection of the voltammetric signal on a pyrolytic graphite electrode. The technique was tested under optimal conditions and various situations limiting or precluding the ligation reaction (such as DNA substrates lacking 5'-phosphate or containing a base mismatch at the nick junction, or application of incompatible cofactor), and utilized for the analysis of the nick-joining activity of a range of recombinant Escherichia coli DNA ligase constructs. The novel technique provides a fast, versatile, specific, and sensitive electrochemical assay of DNA ligase activity.

  3. Characterization and preliminary toxicity assay of nano-titanium dioxide additive in sugar-coated chewing gum.

    Science.gov (United States)

    Chen, Xin-Xin; Cheng, Bin; Yang, Yi-Xin; Cao, Aoneng; Liu, Jia-Hui; Du, Li-Jing; Liu, Yuanfang; Zhao, Yuliang; Wang, Haifang

    2013-05-27

    Nanotechnology shows great potential for producing food with higher quality and better taste through including new additives, improving nutrient delivery, and using better packaging. However, lack of investigations on safety issues of nanofood has resulted in public fears. How to characterize engineered nanomaterials in food and assess the toxicity and health impact of nanofood remains a big challenge. Herein, a facile and highly reliable separation method of TiO2 particles from food products (focusing on sugar-coated chewing gum) is reported, and the first comprehensive characterization study on food nanoparticles by multiple qualitative and quantitative methods is provided. The detailed information on nanoparticles in gum includes chemical composition, morphology, size distribution, crystalline phase, particle and mass concentration, surface charge, and aggregation state. Surprisingly, the results show that the number of food products containing nano-TiO2 (TiO2 in gum is nano-TiO2 , and it is unexpectedly easy to come out and be swallowed by a person who chews gum. Preliminary cytotoxicity assays show that the gum nano-TiO2 particles are relatively safe for gastrointestinal cells within 24 h even at a concentration of 200 μg mL(-1) . This comprehensive study demonstrates accurate physicochemical property, exposure, and cytotoxicity information on engineered nanoparticles in food, which is a prerequisite for the successful safety assessment of nanofood products. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Direct comparison of phosphate uptake by adnate and loosely attached microalgae within an intact biofilm matrix

    International Nuclear Information System (INIS)

    Burkholder, J.M.; Wetzel, R.G.; Klomparens, K.L.

    1990-01-01

    We report a direct comparison of phosphate uptake by adnate and loosely attached microalgae in an intact biofilm matrix, with resolution at the level of individual cells. Track scanning electron microscope autoradiography enabled assay of [ 33 P]phosphate uptake from the overlying water by adnate algae left undisturbed on mature leaves of the macrophyte Potamogeton illinoensis or on artificial plant mimics. The epiphyte communities developed in either phosphate-poor or moderately phosphate-enriched water, and they were assayed on both natural and artificial plants. All adnate taxa examined from both natural and artificial plants in both habitats took up significantly less radiolabel when assayed beneath the overlying matrix than when they were exposed to the water upon removal of the overstory material. Track scanning electron microscope autoradiography and track light microscope autoradiography were intercalibrated to enable comparison of [ 33 P]phosphate uptake by adnate and loosely attached components of the epiphyte matrix. Loosley attached cells on substrata from both habitats took up significantly more radiolabel than did underlying adnate cells, indicating that access to phosphate supplies from the water depended on the position of microbial cells in the matrix. In this short-term assay, the adnate microalgae were relatively isolated from the water column nutrient source

  5. Population screening for glucose-6-phosphate dehydrogenase deficiencies in Isabel Province, Solomon Islands, using a modified enzyme assay on filter paper dried bloodspots

    Directory of Open Access Journals (Sweden)

    Landry Losi

    2010-08-01

    Full Text Available Abstract Background Glucose-6-phosphate dehydrogenase deficiency poses a significant impediment to primaquine use for the elimination of liver stage infection with Plasmodium vivax and for gametocyte clearance, because of the risk of life-threatening haemolytic anaemia that can occur in G6PD deficient patients. Although a range of methods for screening G6PD deficiency have been described, almost all require skilled personnel, expensive laboratory equipment, freshly collected blood, and are time consuming; factors that render them unsuitable for mass-screening purposes. Methods A published WST8/1-methoxy PMS method was adapted to assay G6PD activity in a 96-well format using dried blood spots, and used it to undertake population screening within a malaria survey undertaken in Isabel Province, Solomon Islands. The assay results were compared to a biochemical test and a recently marketed rapid diagnostic test. Results Comparative testing with biochemical and rapid diagnostic test indicated that results obtained by filter paper assay were accurate providing that blood spots were assayed within 5 days when stored at ambient temperature and 10 days when stored at 4 degrees. Screening of 8541 people from 41 villages in Isabel Province, Solomon Islands revealed the prevalence of G6PD deficiency as defined by enzyme activity Conclusions The assay enabled simple and quick semi-quantitative population screening in a malaria-endemic region. The study indicated a high prevalence of G6PD deficiency in Isabel Province and highlights the critical need to consider G6PD deficiency in the context of P. vivax malaria elimination strategies in Solomon Islands, particularly in light of the potential role of primaquine mass drug administration.

  6. A laboratory-scale pretreatment and hydrolysis assay for determination of reactivity in cellulosic biomass feedstocks.

    Science.gov (United States)

    Wolfrum, Edward J; Ness, Ryan M; Nagle, Nicholas J; Peterson, Darren J; Scarlata, Christopher J

    2013-11-14

    The rapid determination of the release of structural sugars from biomass feedstocks is an important enabling technology for the development of cellulosic biofuels. An assay that is used to determine sugar release for large numbers of samples must be robust, rapid, and easy to perform, and must use modest amounts of the samples to be tested.In this work we present a laboratory-scale combined pretreatment and saccharification assay that can be used as a biomass feedstock screening tool. The assay uses a commercially available automated solvent extraction system for pretreatment followed by a small-scale enzymatic hydrolysis step. The assay allows multiple samples to be screened simultaneously, and uses only ~3 g of biomass per sample. If the composition of the biomass sample is known, the results of the assay can be expressed as reactivity (fraction of structural carbohydrate present in the biomass sample released as monomeric sugars). We first present pretreatment and enzymatic hydrolysis experiments on a set of representative biomass feedstock samples (corn stover, poplar, sorghum, switchgrass) in order to put the assay in context, and then show the results of the assay applied to approximately 150 different feedstock samples covering 5 different materials. From the compositional analysis data we identify a positive correlation between lignin and structural carbohydrates, and from the reactivity data we identify a negative correlation between both carbohydrate and lignin content and total reactivity. The negative correlation between lignin content and total reactivity suggests that lignin may interfere with sugar release, or that more mature samples (with higher structural sugars) may have more recalcitrant lignin. The assay presented in this work provides a robust and straightforward method to measure the sugar release after pretreatment and saccharification that can be used as a biomass feedstock screening tool. We demonstrated the utility of the assay by

  7. General and efficient one-pot synthesis of novel sugar/heterocyclic(aryl) 1,2-diketones from sugar terminal alkynes by Sonogashira/tetra-n- butylammonium permanganate oxidation.

    Science.gov (United States)

    Zhang, Fuyi; Wu, Xiaopei; Wang, Liming; Liu, Hong; Zhao, Yufen

    2015-11-19

    A new approach for one-pot synthesis of novel sugar/heterocyclic(aryl) 1,2-diketones has been achieved by the reaction of various sugar terminal alkynes with heterocyclic(aryl) iodides at room temperature. This one-pot protocol includes Sonogashira coupling and mild n-Bu4NMnO4 oxidation reaction. This method is mild, general and efficient. Fifty-six examples have been given and the sugar/heterocyclic(aryl) 1,2-diketones were obtained in 71-94% yields. The sugar terminal alkynes include 9 structurally different sugars in pyranose, furanose, and acyclic form which have various protecting groups, sensitive groups, and sterically bulky substituents. The heterocyclic(aryl) iodides include sterically bulky heterocyclic compounds and iodobenzenes with electron-donating, electron-neutral, and electron-withdrawing substituents. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Evidence for the involvement of Ala 166 in coupling Na(+) to sugar transport through the human Na(+)/glucose cotransporter

    DEFF Research Database (Denmark)

    Meinild, A K; Loo, D D; Hirayama, B A

    2001-01-01

    . The affinity for Na(+) was unchanged compared to that of hSGLT1, whereas the sugar affinity was reduced and sugar specificity was altered. There was a reduction in the turnover rate of the transporter, and in contrast to that of hSGLT1, the turnover rate depended on the sugar molecule. Exposure of A166C......We mutated residue 166, located in the putative Na(+) transport pathway between transmembrane segments 4 and 5 of human Na(+)/glucose cotransporter (hSGLT1), from alanine to cysteine (A166C). A166C was expressed in Xenopus laevis oocytes, and electrophysiological methods were used to assay function...... to MTSEA and MTSET, but not MTSES, abolished sugar transport. Accessibility of A166C to alkylating reagents was independent of protein conformation, indicating that the residue is always accessible from the extracellular surface. Sugar and phlorizin did not protect the residue from being alkylated...

  9. Nonenzymatic gluconeogenesis-like formation of fructose 1,6-bisphosphate in ice.

    Science.gov (United States)

    Messner, Christoph B; Driscoll, Paul C; Piedrafita, Gabriel; De Volder, Michael F L; Ralser, Markus

    2017-07-11

    The evolutionary origins of metabolism, in particular the emergence of the sugar phosphates that constitute glycolysis, the pentose phosphate pathway, and the RNA and DNA backbone, are largely unknown. In cells, a major source of glucose and the large sugar phosphates is gluconeogenesis. This ancient anabolic pathway (re-)builds carbon bonds as cleaved in glycolysis in an aldol condensation of the unstable catabolites glyceraldehyde 3-phosphate and dihydroxyacetone phosphate, forming the much more stable fructose 1,6-bisphosphate. We here report the discovery of a nonenzymatic counterpart to this reaction. The in-ice nonenzymatic aldol addition leads to the continuous accumulation of fructose 1,6-bisphosphate in a permanently frozen solution as followed over months. Moreover, the in-ice reaction is accelerated by simple amino acids, in particular glycine and lysine. Revealing that gluconeogenesis may be of nonenzymatic origin, our results shed light on how glucose anabolism could have emerged in early life forms. Furthermore, the amino acid acceleration of a key cellular anabolic reaction may indicate a link between prebiotic chemistry and the nature of the first metabolic enzymes.

  10. Fatty acid synthase cooperates with glyoxalase 1 to protect against sugar toxicity.

    Directory of Open Access Journals (Sweden)

    Damien Garrido

    2015-02-01

    Full Text Available Fatty acid (FA metabolism is deregulated in several human diseases including metabolic syndrome, type 2 diabetes and cancers. Therefore, FA-metabolic enzymes are potential targets for drug therapy, although the consequence of these treatments must be precisely evaluated at the organismal and cellular levels. In healthy organism, synthesis of triacylglycerols (TAGs-composed of three FA units esterified to a glycerol backbone-is increased in response to dietary sugar. Saturation in the storage and synthesis capacity of TAGs is associated with type 2 diabetes progression. Sugar toxicity likely depends on advanced-glycation-end-products (AGEs that form through covalent bounding between amine groups and carbonyl groups of sugar or their derivatives α-oxoaldehydes. Methylglyoxal (MG is a highly reactive α-oxoaldehyde that is derived from glycolysis through a non-enzymatic reaction. Glyoxalase 1 (Glo1 works to neutralize MG, reducing its deleterious effects. Here, we have used the power of Drosophila genetics to generate Fatty acid synthase (FASN mutants, allowing us to investigate the consequence of this deficiency upon sugar-supplemented diets. We found that FASN mutants are lethal but can be rescued by an appropriate lipid diet. Rescued animals do not exhibit insulin resistance, are dramatically sensitive to dietary sugar and accumulate AGEs. We show that FASN and Glo1 cooperate at systemic and cell-autonomous levels to protect against sugar toxicity. We observed that the size of FASN mutant cells decreases as dietary sucrose increases. Genetic interactions at the cell-autonomous level, where glycolytic enzymes or Glo1 were manipulated in FASN mutant cells, revealed that this sugar-dependent size reduction is a direct consequence of MG-derived-AGE accumulation. In summary, our findings indicate that FASN is dispensable for cell growth if extracellular lipids are available. In contrast, FA-synthesis appears to be required to limit a cell

  11. Production of biomass/energy crops on phosphatic clay soils in central Florida

    Energy Technology Data Exchange (ETDEWEB)

    Stricker, J.A. [Univ. of Florida, Bartow, FL (United States); Prine, G.M.; Woodard, K.R. [Univ. of Florida, Gainesville, FL (United States); Anderson, D.L. [Univ. of Florida, Belle Glade, FL (United States); Shibles, D.B.; Riddle, T.C. [Mined Lands Agricultural Research/Demonstration Project, Bartow, FL (United States)

    1993-12-31

    Phosphatic clay is a byproduct of phosphate mining. Presently more than 40,470 ha have been created, most in central Florida, and about 810 ha are being added each year. Phosphatic clays have high fertility and high water holding capacity, reducing fertilization costs and producing high yields without irrigation. Based on 10 years of research, scientists have selected tall annual-regenerating perennial C-4 grasses as having the greatest potential for biomass production in Florida. The purpose of this work was to determine the feasibility of growing these tall perennial grasses for biomass on phosphatic clay. Elephantgrass, sugarcane and energycane, and erianthus were planted in duplicate replications on phosphatic clay soil in late August, 1986. yield was measured by one harvest in December or January each year for four years. Nitrogen fertilization included 112 kg ha{sup {minus}1} the first year followed by 134 kg ha{sup {minus}1} for the next three years. Nitrogen is the only supplemental nutrient needed to grow all tall grass crops on phosphatic clay. The average annual oven dry matter yield over the 4-yr period was 36.3 Mg ha{sup {minus}1} for PI 300086 elephantgrass, 45.2 for N51 elephantgrass, 42.5 for L79-1002 energycane, 49.0 for US72-1153 energycane, 49.7 for US78-1009 sugarcane, 52.2 for US56-9 sugarcane, 56.2 for CP72-1210 sugarcane, and 48.8 for 1K-7647 erianthus. More recent work has utilized domestic sewage sludge as a nitrogen source for the tall grasses. Preliminary sugar yields of selected sugarcane accessions & sweet sorghum were 4.7 Mg ha{sup {minus}1} for CP72-1210, 12.5 for US67-2022, 3.4 for US78-1009 and 1.3 Mg ha{sup {minus}1} for sweet sorghum. The high yields of the tall grasses grown on phosphatic clay with low inputs indicate a great potential for these crops as a source of renewable energy. A sustainable cropping system may be maintained by utilizing municipal sewage sludge as a nitrogen source with tall grasses on phosphatic clay.

  12. Free sugar contents in onion bulbs on different cultivars and different production areas, and their changes by storage and gamma-irradiation. Free sugars in onion bulbs, 1

    Energy Technology Data Exchange (ETDEWEB)

    Nishibori, Sukie [Tokaigakuen Women' s Coll., Nagoya (Japan); Namiki, Kazuko

    1982-05-01

    Changes in the contents of free sugar in onion bulbs of different cultivars and different production areas were investigated during storage and processing by using gas-chromatography. No significant difference was observed in the total free sugar contents among the onions of different cultivars and different production areas except early season's cultivars which showed a slightly lower content of sugar. The free sugar was mainly composed with glucose (1.7 - 3.2%), fructose (1.1 - 2.5%) and sucrose (0.7 - 2.6%), and the composition was somewhat different among the onions of different cultivars and different production areas. During the storage at 4 - 5/sup 0/C, the total free suger contents gradually decreased during the first few months but more markedly to about 30% after sprouting. The middle part of onion bulbs contained more total free sugar than the other parts (middle part, 5.6 ..-->.. 3.5%, inner part, 5.2 ..-->.. 3.2%, outer part, 4.8 ..-->.. 1.5%), and the total free sugar contents in the outer part decreased markedly during the storage. The changes of the sugar contents after the freeze-drying were almost negligible, but showed significant decrease after the hot-air drying, especially for sucrose. The gamma irradiation with 5 - 50 krad gave no appreciable effects on the total free sugar contents though it seemed to make a slight increase in the sucrose content.

  13. The Path of Carbon in Photosynthesis XIX. The Identification of Sucrose Phosphate in Sugar Beet Leaves

    Science.gov (United States)

    Buchanan, J. G.

    1952-09-01

    The recognition and characterization of a sucrose phosphate as an intermediate in sucrose by synthesis by green plants is described. A tentative structure for this phosphate is proposed and its mode of formation suggested.

  14. Sphingosine-1-phosphate signaling in the cardiovascular system

    NARCIS (Netherlands)

    Peters, Stephan L. M.; Alewijnse, Astrid E.

    2007-01-01

    Increasing evidence suggests a key role for the bioactive lipid sphingosine-1-phosphate (S1P) and its G-protein-coupled receptors (S1P(1-5)) in the cardiovascular system. Recent advances in sphingolipid research indicates that cardiomyocyte, vascular smooth muscle and endothelial cell function is

  15. Regulation of Soluble Phosphate on the Ability of Phytate Mineralization and β-Propeller Phytase Gene Expression of Pseudomonas fluorescens JZ-DZ1, a Phytate-Mineralizing Rhizobacterium.

    Science.gov (United States)

    Shen, Lan; Wu, Xiao-Qin; Zeng, Qing-Wei; Liu, Hong-Bin

    2016-12-01

    Phytate-mineralizing rhizobacteria (PMR) play an important role in providing phosphorus for the sustainable plant growth. It is important to investigate the ability of PMR to produce phytase under different phosphate levels for its application. The effects of different concentrations of soluble phosphate on the ability of phytate mineralization of Pseudomonas fluorescens JZ-DZ1, a phytate-mineralizing rhizobacterium, were investigated in both solid and liquid media. The results on solid media showed that halo zone width gradually reduced with concentrations of soluble phosphate increasing from 0.05 to 20 mM, indicating the reduction of the ability of phytate mineralization. The results were consistent with the quantitative detection of phytase activity from the overall trend. An 1866-bp β-propeller phytase (BPP) gene (phyPf) was cloned from the strain, and the deduced amino acid sequence of phyPf shared 98 % of identity with a known BPP from Pseudomonas sp. BS10-3 (AJF36073.1). The results of relative real-time quantitative PCR assay showed that the expression of phyPf was induced by a low concentration (0.1 mM) of soluble phosphate, suggesting that BPP secretion was regulated by gene phyPf. The BPP-harboring bacterium P. fluorescens JZ-DZ1 with low phosphate-inducible ability of phytate mineralization could be potentially applied to promote phosphorus uptake for plants in the future.

  16. RESEARCH OF BIOTRANSFORMATION PROCESS OF CONTAINING SUGAR RAW FOR ISOMALTULOSE RECEIVING

    Directory of Open Access Journals (Sweden)

    O. S. Korneeva

    2014-01-01

    Full Text Available The process of biotransformation containing sugar raw isomaltulosesynthase bacteria of the genus Erwinia to produce isomaltulose - natural sugars substitute. Raw cane sugar, beet molasses and sweet sorghum syrup used for isomaltulose. It was established that cane sugar may serve as a substrate for the enzymatic reaction together with biotransformation of pure sucrose. Yield of isomaltulose in the transformation of raw sugar was comparable to the control and was 97% for 3,5 hours at the optimal isomerization conditions (pH 6,0, 30 º C, the enzyme dosage of 5 U / mg of sucrose. Scientifically substantiated reasons for reducing the degree of transformation of molasses, sugar syrup strength sorghum. The influence of non-sugars accompanying basic substrates - raw cane sugar, molasses, sweet sorghum syrup, when biocatalytic transformation. Proved inhibitory effect of metal ion (aluminum , iron , anions of inorganic acids ( nitrate , chloride, phosphate , amino acids (serine , aspartic acid -hand , organic acids ( citric acid, acetic acid, the process of biotransformation , the formation of sucrose. Reducing the degree of isomerization in the presence of sucrose was observed in the study of non-sugars from 50 to 65 % compared with the control. Isomaltulosesynthase activating effect on , and consequently , the yield of isomaltulose to set calcium and manganese in the form of their sulfates. The introduction of these components into a solution of pure sucrose increases the yield of isomaltulose by 30.5 and 13.2 % respectively. The data obtained will be the basis of studies to optimize the process of biotransformation of various sources of vegetable raw materials rich in sucrose to produce isomaltulose.

  17. Sugar cane stillage: a potential source of natural antioxidants.

    Science.gov (United States)

    Caderby, Emma; Baumberger, Stéphanie; Hoareau, William; Fargues, Claire; Decloux, Martine; Maillard, Marie-Noëlle

    2013-11-27

    Biorefinery of sugar cane is the first economic activity of Reunion Island. Some sugar cane manufactured products (juice, syrup, molasses) have antioxidant activities and are sources of both phenolic compounds and Maillard Reaction Products (MRP). The study aimed to highlight the global antioxidant activity of sugar cane stillage and understand its identity. Chromatographic fractionation on Sephadex LH-20 resin allowed the recovery of a MRP-rich fraction, responsible for 58 to 66% of the global antioxidant activity according to the nature of the sugar cane stillage (DPPH test), and a phenolic compounds-rich fraction for 37 to 59% of the activity. A good correlation was recorded between the antioxidant activity of the sugar cane stillage and its content in total reducing compounds amount (Folin-Ciocalteu assay), among them 2.8 to 3.9 g/L of phenolic compounds (in 5-caffeoylquinic acid equivalent). Preliminary experiments by HPLC-DAD-MS allowed to identify several free phenolic acids and gave clues to identify esters of quinic acids.

  18. Novel mode of inhibition by D-tagatose 6-phosphate through a Heyns rearrangement in the active site of transaldolase B variants.

    Science.gov (United States)

    Stellmacher, Lena; Sandalova, Tatyana; Schneider, Sarah; Schneider, Gunter; Sprenger, Georg A; Samland, Anne K

    2016-04-01

    Transaldolase B (TalB) and D-fructose-6-phosphate aldolase A (FSAA) from Escherichia coli are C-C bond-forming enzymes. Using kinetic inhibition studies and mass spectrometry, it is shown that enzyme variants of FSAA and TalB that exhibit D-fructose-6-phosphate aldolase activity are inhibited covalently and irreversibly by D-tagatose 6-phosphate (D-T6P), whereas no inhibition was observed for wild-type transaldolase B from E. coli. The crystal structure of the variant TalB(F178Y) with bound sugar phosphate was solved to a resolution of 1.46 Å and revealed a novel mode of covalent inhibition. The sugar is bound covalently via its C2 atom to the ℇ-NH2 group of the active-site residue Lys132. It is neither bound in the open-chain form nor as the closed-ring form of D-T6P, but has been converted to β-D-galactofuranose 6-phosphate (D-G6P), a five-membered ring structure. The furanose ring of the covalent adduct is formed via a Heyns rearrangement and subsequent hemiacetal formation. This reaction is facilitated by Tyr178, which is proposed to act as acid-base catalyst. The crystal structure of the inhibitor complex is compared with the structure of the Schiff-base intermediate of TalB(E96Q) formed with the substrate D-fructose 6-phosphate determined to a resolution of 2.20 Å. This comparison highlights the differences in stereochemistry at the C4 atom of the ligand as an essential determinant for the formation of the inhibitor adduct in the active site of the enzyme.

  19. Soil Chemical Properties and Nutrient Uptake of Cocoa as Affected by Application of Different Organic Matters and Phosphate Fertilizers

    Directory of Open Access Journals (Sweden)

    Sugiyanto Sugiyanto

    2008-07-01

    Full Text Available Effort repair of land quality better be done by simultan namely with application of organic matters and inorganic fertilization. The objective of this research is to study the effect of varied organic matters source and phosphate fertilizers on the chemicals soil characteristic and cocoa nutrient uptake. The experiment was laid experimentally in split-plot design and environmentally in randomized complete block design. The main plot was source of P consisted of, control, SP 36 and rock phosphate in dosage of 200 mg P2O5 per kg of air dry soil. Source of organic matter as sub-plot consisted of control (no organic matter, cow dung, cocoa pod husk compost and sugar cane filter cake, each in dosage of 2.5 and 5.0%. Result of this experiment showed application of cow dung, cocoa pod husk compost and sugar cane filter cake increased content of C, N, Ca exchangeable, Fe available, and pH in soil, and SP 36 increased availability of P in soil. Application of sugar cane filter cake increased N, K, Ca, Mg, and SO4 uptake but did not increase Cl uptake, application of cow dung in dosage 5% increased N, K, and Cl uptake and cocoa pod husk compost dosage 5% increased N and K uptake of cocoa. SP 36 increased Mg uptake of cocoa but rock phosphate did not increase it. They were not interaction between organic matters and phosphate fertilizers to nutrient uptake of cocoa. Nutrient soil content as affected by organic matters correlated with nutrient uptake of cocoa.Key words : soil chemical properties, nutrient uptake, cocoa, organic matter, phosphate fertlizers.

  20. Arabidopsis scaffold protein RACK1A modulates rare sugar D-allose regulated gibberellin signaling

    OpenAIRE

    Fennell, Herman; Olawin, Abdulquadri; Mizanur, Rahman M.; Izumori, Ken; Chen, Jin-Gui; Ullah, Hemayet

    2012-01-01

    As energy sources and structural components, sugars are the central regulators of plant growth and development. In addition to the abundant natural sugars in plants, more than 50 different kinds of rare sugars exist in nature, several of which show distinct roles in plant growth and development. Recently, one of the rare sugars, D-allose, an epimer of D-glucose at C3, is found to suppress plant hormone gibberellin (GA) signaling in rice. Scaffold protein RACK1A in the model plant Arabidopsis ...

  1. The Effects of Fenarimol and Methyl Parathion on Glucose 6-Phosphate Dehydrogenase Enzyme Activity in Rats

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    Ferda ARI

    2017-10-01

    Full Text Available Fenarimol and methyl parathion are pesticides that have been used in agriculture for several years. These pesticides have significant effects on environmental and human health. Therefore, we investigated the effects of methyl parathion and fenarimol on glucose 6-phosphate dehydrogenase (EC 1.1.1.49 enzyme activity in rats. The glucose 6- phosphate dehydrogenase is the first enzyme of the pentose phosphate pathway and it is important in detoxifying reactions by NADPH generated. In this study, wistar albino rats administrated with methyl parathion (7 mg kg–1 and fenarimol (200 mg kg−1 by intraperitoneally for different periods (2, 4, 8, 16, 32, 64, and 72 h. The glucose 6-phosphate dehydrogenase enzyme activity was assayed in liver, kidney, brain, and small intestine in male and female rats. The exposure of fenarimol and methyl parathion caused increase of glucose 6-phosphate dehydrogenase enzyme activity in rat tissues, especially at last periods. We suggest that this increment of enzyme activity may be the reason of toxic effects of fenarimol and methyl parathion.

  2. Arabidopsis scaffold protein RACK1A modulates rare sugar D-allose regulated gibberellin signaling.

    Science.gov (United States)

    Fennell, Herman; Olawin, Abdulquadri; Mizanur, Rahman M; Izumori, Ken; Chen, Jin-Gui; Ullah, Hemayet

    2012-11-01

    As energy sources and structural components, sugars are the central regulators of plant growth and development. In addition to the abundant natural sugars in plants, more than 50 different kinds of rare sugars exist in nature, several of which show distinct roles in plant growth and development. Recently, one of the rare sugars, D-allose, an epimer of D-glucose at C3, is found to suppress plant hormone gibberellin (GA) signaling in rice. Scaffold protein RACK1A in the model plant Arabidopsis is implicated in the GA pathway as rack1a knockout mutants show insensitivity to GA in GA-induced seed germination. Using genetic knockout lines and a reporter gene, the functional role of RACK1A in the D-allose pathway was investigated. It was found that the rack1a knockout seeds showed hypersensitivity to D-allose-induced inhibition of seed germination, implicating a role for RACK1A in the D-allose mediated suppression of seed germination. On the other hand, a functional RACK1A in the background of the double knockout mutations in the other two RACK1 isoforms, rack1b/rack1c, showed significant resistance to the D-allose induced inhibition of seed germination. The collective results implicate the RACK1A in the D-allose mediated seed germination inhibition pathway. Elucidation of the rare sugar signaling mechanism will help to advance understanding of this less studied but important cellular signaling pathway.

  3. Biocompatibility of the titanium-based implant surfaces: Effect of the calcium dihydrogen phosphate on osteoblast cells

    Directory of Open Access Journals (Sweden)

    Kaluđerović Milena R.

    2016-01-01

    Full Text Available The influence of the presence of calcium dihydrogen phosphate in acid media on titanium-based implant surfaces, Ticer, employed in clinics, and its white form (Ticer white, on osteoblast cells was investigated. Novel surfaces M1 and M2 were obtained by immersing Ticer and Ticer white surfaces in calcium dihydrogen phosphate solution at pH 3.5. The surfaces were characterized by SEM, EDS and X-ray diffraction. The results related to interaction of investigated surfaces and human osteoblast cells from indirect biocompatibility (MTT and SRB assays, proliferation (DAPI assay and mode of cell death (acridine orange/ethidium bromide (AO/EB double staining were found to be in good agreement, as well as findings from osteocalcin (OC and bone sialoprotein (BSP expression. Surfaces were obtained by employing anodic plasma-electrochemical oxidation with spark discharges without subsequent surface modifications were found to be more compatible. Soaking of Ticer and Ticer white in phosphate solution gave toxic materials (M1 and M2 which induced apoptosis and secondary necrosis in osteoblast cells.

  4. Regulation of root morphogenesis in arbuscular mycorrhizae: what role do fungal exudates, phosphate, sugars and hormones play in lateral root formation?

    Science.gov (United States)

    Fusconi, Anna

    2014-01-01

    Background Arbuscular mycorrhizae (AMs) form a widespread root–fungus symbiosis that improves plant phosphate (Pi) acquisition and modifies the physiology and development of host plants. Increased branching is recognized as a general feature of AM roots, and has been interpreted as a means of increasing suitable sites for colonization. Fungal exudates, which are involved in the dialogue between AM fungi and their host during the pre-colonization phase, play a well-documented role in lateral root (LR) formation. In addition, the increased Pi content of AM plants, in relation to Pi-starved controls, as well as changes in the delivery of carbohydrates to the roots and modulation of phytohormone concentration, transport and sensitivity, are probably involved in increasing root system branching. Scope This review discusses the possible causes of increased branching in AM plants. The differential root responses to Pi, sugars and hormones of potential AM host species are also highlighted and discussed in comparison with those of the non-host Arabidopsis thaliana. Conclusions Fungal exudates are probably the main compounds regulating AM root morphogenesis during the first colonization steps, while a complex network of interactions governs root development in established AMs. Colonization and high Pi act synergistically to increase root branching, and sugar transport towards the arbusculated cells may contribute to LR formation. In addition, AM colonization and high Pi generally increase auxin and cytokinin and decrease ethylene and strigolactone levels. With the exception of cytokinins, which seem to regulate mainly the root:shoot biomass ratio, these hormones play a leading role in governing root morphogenesis, with strigolactones and ethylene blocking LR formation in the non-colonized, Pi-starved plants, and auxin inducing them in colonized plants, or in plants grown under high Pi conditions. PMID:24227446

  5. Radiometric assays for glycerol, glucose, and glycogen

    International Nuclear Information System (INIS)

    Bradley, D.C.; Kaslow, H.R.

    1989-01-01

    We have developed radiometric assays for small quantities of glycerol, glucose and glycogen, based on a technique described by Thorner and Paulus for the measurement of glycerokinase activity. In the glycerol assay, glycerol is phosphorylated with [32P]ATP and glycerokinase, residual [32P]ATP is hydrolyzed by heating in acid, and free [32P]phosphate is removed by precipitation with ammonium molybdate and triethylamine. Standard dose-response curves were linear from 50 to 3000 pmol glycerol with less than 3% SD in triplicate measurements. Of the substances tested for interference, only dihydroxyacetone gave a slight false positive signal at high concentration. When used to measure glycerol concentrations in serum and in media from incubated adipose tissue, the radiometric glycerol assay correlated well with a commonly used spectrophotometric assay. The radiometric glucose assay is similar to the glycerol assay, except that glucokinase is used instead of glycerokinase. Dose response was linear from 5 to 3000 pmol glucose with less than 3% SD in triplicate measurements. Glucosamine and N-acetylglucosamine gave false positive signals when equimolar to glucose. When glucose concentrations in serum were measured, the radiometric glucose assay agreed well with hexokinase/glucose-6-phosphate dehydrogenase (H/GDH)-based and glucose oxidase/H2O2-based glucose assays. The radiometric method for glycogen measurement incorporates previously described isolation and digestion techniques, followed by the radiometric assay of free glucose. When used to measure glycogen in mouse epididymal fat pads, the radiometric glycogen assay correlated well with the H/GDH-based glycogen assay. All three radiometric assays offer several practical advantages over spectral assays

  6. Influence of calcium on ceramide-1-phosphate monolayers

    Directory of Open Access Journals (Sweden)

    Joana S. L. Oliveira

    2016-02-01

    Full Text Available Ceramide-1-phosphate (C1P plays an important role in several biological processes, being identified as a key regulator of many protein functions. For instance, it acts as a mediator of inflammatory responses. The mediation of the inflammation process happens due to the interaction of C1P with the C2 domain of cPLA2α, an effector protein that needs the presence of submicromolar concentrations of calcium ions. The aim of this study was to determine the phase behaviour and structural properties of C1P in the presence and absence of millimolar quantities of calcium in a well-defined pH environment. For that purpose, we used monomolecular films of C1P at the soft air/liquid interface with calcium ions in the subphase. The pH was varied to change the protonation degree of the C1P head group. We used surface pressure versus molecular area isotherms coupled with other monolayer techniques as Brewster angle microscopy (BAM, infrared reflection–absorption spectroscopy (IRRAS and grazing incidence X-ray diffraction (GIXD. The isotherms indicate that C1P monolayers are in a condensed state in the presence of calcium ions, regardless of the pH. At higher pH without calcium ions, the monolayer is in a liquid-expanded state due to repulsion between the negatively charged phosphate groups of the C1P molecules. When divalent calcium ions are added, they are able to bridge the highly charged phosphate groups, enhancing the regular arrangement of the head groups. Similar solidification of the monolayer structure can be seen in the presence of a 150 times larger concentration of monovalent sodium ions. Therefore, calcium ions have clearly a strong affinity for the phosphomonoester of C1P.

  7. Aspirin Inhibits Platelet-Derived Sphingosine-1-Phosphate Induced Endothelial Cell Migration.

    Science.gov (United States)

    Polzin, Amin; Knoop, Betül; Böhm, Andreas; Dannenberg, Lisa; Zurek, Mark; Zeus, Tobias; Kelm, Malte; Levkau, Bodo; Rauch, Bernhard H

    2018-01-01

    Aspirin plays a crucial role in the prevention of cardiovascular diseases. We previously described that aspirin has effects beyond inhibition of platelet aggregation, as it inhibited thrombin-mediated release of sphingosine-1-phosphate (S1P) from human platelets. S1P is a bioactive lipid with important functions on inflammation and apoptosis. In endothelial cells (EC), S1P is a key regulator of cell migration. In this study, we aimed to analyze the effects of aspirin on platelet-induced EC migration. Human umbilical EC migration was measured by Boyden chamber assay. EC migration was induced by platelet supernatants of thrombin receptor-activating peptide-1 (AP1) stimulated platelets. To investigate the S1P receptor subtype that promotes EC migration, specific inhibitors of S1P receptor subtypes were applied. S1P induced EC migration in a concentration-dependent manner. EC migration induced by AP1-stimulated platelet supernatants was reduced by aspirin. S1P1 receptor inhibition almost completely abolished EC migration induced by activated platelets. The inhibition of S1P2 or S1P3 receptor had no effect. Aspirin inhibits EC migration induced by activated platelets that is in part due to S1P and mediated by the endothelial S1P1 receptor. The clinical significance of this novel mechanism of aspirin action has to be investigated in future studies. © 2017 S. Karger AG, Basel.

  8. NELL-1 increases pre-osteoblast mineralization using both phosphate transporter Pit1 and Pit2

    Energy Technology Data Exchange (ETDEWEB)

    Cowan, Catherine M. [Department of Bioengineering, University of California, Los Angeles, 420 Westwood Plaza,7523 Boelter Hall, Los Angeles, CA 90095 (United States); Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, University of California, Los Angeles, 40833 Le Conte Ave, Los Angeles, CA 90095 (United States); Zhang, Xinli; James, Aaron W.; Mari Kim, T.; Sun, Nichole [Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, University of California, Los Angeles, 40833 Le Conte Ave, Los Angeles, CA 90095 (United States); Wu, Benjamin [Department of Bioengineering, University of California, Los Angeles, 420 Westwood Plaza,7523 Boelter Hall, Los Angeles, CA 90095 (United States); Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, University of California, Los Angeles, 40833 Le Conte Ave, Los Angeles, CA 90095 (United States); Ting, Kang [Dental and Craniofacial Research Institute and Section of Orthodontics, School of Dentistry, University of California, Los Angeles, 40833 Le Conte Ave, Los Angeles, CA 90095 (United States); Soo, Chia, E-mail: bsoo@ucla.edu [UCLA and Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic, Hospital Research Center, University of California, Los Angeles, 2641 Charles E. Young Dr. South, Los Angeles, CA 90095 (United States)

    2012-06-08

    Highlights: Black-Right-Pointing-Pointer NELL-1 accelerates extracellular matrix mineralization in MC3T3-E1 pre-osteoblasts. Black-Right-Pointing-Pointer NELL-1 significantly increases intracellular inorganic phosphate levels. Black-Right-Pointing-Pointer NELL-1 positively regulates osteogenesis but not proliferation in MC3T3-E1 cells. Black-Right-Pointing-Pointer NELL-1 regulates inorganic phosphate transporter activity. -- Abstract: NELL-1 is a potent osteoinductive molecule that enhances bone formation in multiple animal models through currently unidentified pathways. In the present manuscript, we hypothesized that NELL-1 may regulate osteogenic differentiation accompanied by alteration of inorganic phosphate (Pi) entry into the osteoblast via sodium dependent phosphate (NaPi) transporters. To determine this, MC3T3-E1 pre-osteoblasts were cultured in the presence of recombinant human (rh)NELL-1 or rhBMP-2. Analysis was performed for intracellular Pi levels through malachite green staining, Pit-1 and Pit-2 expression, and forced upregulation of Pit-1 and Pit-2. Results showed rhNELL-1 to increase MC3T3-E1 matrix mineralization and Pi influx associated with activation of both Pit-1 and Pit-2 channels, with significantly increased Pit-2 production. In contrast, Pi transport elicited by rhBMP-2 showed to be associated with increased Pit-1 production only. Next, neutralizing antibodies against Pit-1 and Pit-2 completely abrogated the Pi influx effect of rhNELL-1, suggesting rhNELL-1 is dependent on both transporters. These results identify one potential mechanism of action for rhNELL-1 induced osteogenesis and highlight a fundamental difference between NELL-1 and BMP-2 signaling.

  9. NELL-1 increases pre-osteoblast mineralization using both phosphate transporter Pit1 and Pit2

    International Nuclear Information System (INIS)

    Cowan, Catherine M.; Zhang, Xinli; James, Aaron W.; Mari Kim, T.; Sun, Nichole; Wu, Benjamin; Ting, Kang; Soo, Chia

    2012-01-01

    Highlights: ► NELL-1 accelerates extracellular matrix mineralization in MC3T3-E1 pre-osteoblasts. ► NELL-1 significantly increases intracellular inorganic phosphate levels. ► NELL-1 positively regulates osteogenesis but not proliferation in MC3T3-E1 cells. ► NELL-1 regulates inorganic phosphate transporter activity. -- Abstract: NELL-1 is a potent osteoinductive molecule that enhances bone formation in multiple animal models through currently unidentified pathways. In the present manuscript, we hypothesized that NELL-1 may regulate osteogenic differentiation accompanied by alteration of inorganic phosphate (Pi) entry into the osteoblast via sodium dependent phosphate (NaPi) transporters. To determine this, MC3T3-E1 pre-osteoblasts were cultured in the presence of recombinant human (rh)NELL-1 or rhBMP-2. Analysis was performed for intracellular Pi levels through malachite green staining, Pit-1 and Pit-2 expression, and forced upregulation of Pit-1 and Pit-2. Results showed rhNELL-1 to increase MC3T3-E1 matrix mineralization and Pi influx associated with activation of both Pit-1 and Pit-2 channels, with significantly increased Pit-2 production. In contrast, Pi transport elicited by rhBMP-2 showed to be associated with increased Pit-1 production only. Next, neutralizing antibodies against Pit-1 and Pit-2 completely abrogated the Pi influx effect of rhNELL-1, suggesting rhNELL-1 is dependent on both transporters. These results identify one potential mechanism of action for rhNELL-1 induced osteogenesis and highlight a fundamental difference between NELL-1 and BMP-2 signaling.

  10. A clinical study of serum phosphate and magnesium in type II diabetes mellitus

    Directory of Open Access Journals (Sweden)

    Revathi.R, Julius Amaldas

    2014-11-01

    Full Text Available To assess serum phosphate and magnesium level in type-2 diabetic patients in comparison with those of control subjects. Methodology: There were 100 diabetic patients and 100 age matched non-diabetic (control subjects included in this study. Serum phosphate, serum magnesium and fasting and postprandial blood sugar measured among the diabetic and control groups using SPSS version 16.0 for analysis. Results: Serum phosphate level was significantly lower in diabetic patients (2.92 ± 0.75 as compared to control subjects (3.38 ± 0.49. Serum magnesium levels were significantly lower in diabetic patients (0.9 ± 0.15 compared to controls (2.75 ± 0.46 Conclusion: The study reveals that hyperglycemia may reduce serum levels of magnesium and phosphorus.

  11. Effect of different levels of sugar on qualitative characteristics of lassi prepared from sour dahi

    Directory of Open Access Journals (Sweden)

    Muhammad Shah Moazzem

    2018-08-01

    Full Text Available An investigation was carried out to develop lassi from sour dahi using different levels of sugar (10, 15, 20 and 25% and 15% water. Lassi quality was assayed through the study of physical, chemical and microbiological parameters. Results revealed that significant difference existed in overall physical score of lassi samples and the highest score was found in 15% sugar lassi whereas, the lowest score was found in 25% sugar lassi. Total solids, carbohydrate, fat, protein and ash contents differed significantly among various levels of sugar added lassi. From chemical test, it appears that, 15% sugar added lassi possess the highest fat and protein values whereas, the highest total solids and carbohydrate values posses in 25% sugar added lassi. No significant difference (p>0.05 revealed in terms of pH value and acidity percentage among lassi types. Lassi made from 10% sugar was most inferior than other levels of sugar added lassi in respect of microbiological quality- total viable count (×104 cfu/mL content was 95.67±2.08 and coliform (×10 cfu/mL content was 1.00±0.00. Considering above mentioned quality aspects, it might be resolved that lassi could be prepared successfully from sour dahi with 15% sugar keeping water level constant at 15%. [Fundam Appl Agric 2018; 3(2.000: 434-439

  12. Sugar dosimeters. Part 1. State of the art

    International Nuclear Information System (INIS)

    Peimel-Stuglik, Z.

    2008-01-01

    A review of the literature dealing with the possibility of using sugars, in particular sucrose, as dosimetric material is presented. All methods involved were divided according to analytical techniques used in dosimetric signal measurements (polarimetry, spectrophotometry and electron paramagnetic resonance - EPR). Double-signal sugar dosimeters (EPR + spectrophotometry) are also described. (author) [pl

  13. ASP4058, a novel agonist for sphingosine 1-phosphate receptors 1 and 5, ameliorates rodent experimental autoimmune encephalomyelitis with a favorable safety profile.

    Directory of Open Access Journals (Sweden)

    Rie Yamamoto

    Full Text Available Sphingosine-1-phosphate (S1P is a biologically active sphingolipid that acts through the members of a family of five G protein-coupled receptors (S1P1-S1P5. S1P1 is a major regulator of lymphocyte trafficking, and fingolimod, whose active metabolite fingolimod phosphate acts as a nonselective S1P receptor agonist, exerts its immunomodulatory effect, at least in part, by regulating the lymphocyte trafficking by inducing down regulation of lymphocyte S1P1. Here, we detail the pharmacological profile of 5-{5-[3-(trifluoromethyl-4-{[(2S-1,1,1-trifluoropropan-2-yl]oxy}phenyl]-1,2,4-oxadiazol-3-yl}-1H-benzimidazole (ASP4058, a novel next-generation S1P receptor agonist selective for S1P1 and S1P5. ASP4058 preferentially activates S1P1 and S1P5 compared with S1P2, 3, 4 in GTPγS binding assays in vitro. Oral administration of ASP4058 reduced the number of peripheral lymphocytes and inhibited the development of experimental autoimmune encephalomyelitis (EAE in Lewis rats. Further, ASP4058 prevented relapse of disease in a mouse model of relapsing-remitting EAE. Although these immunomodulatory effects were comparable to those of fingolimod, ASP4058 showed a wider safety margin than fingolimod for bradycardia and bronchoconstriction in rodents. These observations suggest that ASP4058 represents a new therapeutic option for treating multiple sclerosis that is safer than nonselective S1P receptor agonists such as fingolimod.

  14. Sphingosine-1-phosphate stimulates rat primary chondrocyte proliferation

    International Nuclear Information System (INIS)

    Kim, Mi-Kyoung; Lee, Ha Young; Kwak, Jong-Young; Park, Joo-In; Yun, Jeanho; Bae, Yoe-Sik

    2006-01-01

    Rat primary chondrocytes express the sphingosine-1-phosphate (S1P) receptor, S1P 2 , S1P 3 , S1P 4 , but not S1P 1 . When chondrocytes were stimulated with S1P or phytosphingosine-1-phosphate (PhS1P, an S1P 1 - and S1P 4 -selective agonist), phospholipase C-mediated cytosolic calcium increase was dramatically induced. S1P and PhS1P also stimulated two kinds of mitogen-activated protein kinases, extracellular signal-regulated kinase (ERK) and p38 kinase in chondrocytes. In terms of the two phospholipids-mediated functional modulation of chondrocytes, S1P and PhS1P stimulated cellular proliferation. The two phospholipids-induced chondrocyte proliferations were almost completely blocked by PD98059 but not by SB203580, suggesting that ERK but not p38 kinase is essentially required for the proliferation. Pertussis toxin almost completely inhibited the two phospholipids-induced cellular proliferation and ERK activation, indicating the crucial role of G i protein. This study demonstrates the physiological role of two important phospholipids (S1P and PhS1P) on the modulation of rat primary chondrocyte proliferation, and the crucial role played by ERK in the process

  15. Regulation of fruit and seed response to heat and drought by sugars as nutrients and signals

    Directory of Open Access Journals (Sweden)

    Yong-Hua eLiu

    2013-08-01

    Full Text Available A large body of evidence shows that sugars function both as nutrients and signals to regulate fruit and seed set under normal and stress conditions including heat and drought. Inadequate sucrose import to, and its degradation within, reproductive organs cause fruit and seed abortion under heat and drought. As nutrients, sucrose-derived hexoses provide carbon skeletons and energy for growth and development of fruits and seeds. Sugar metabolism can also alleviate the impact of stress on fruit and seed through facilitating biosynthesis of heat shock proteins (Hsps and non-enzymic antioxidants (e.g. glutathione, ascorbic acid, which collectively maintain the integrity of membranes and prevent programmed cell death (PCD through protecting proteins and scavenging reactive oxygen species (ROS. In parallel, sugars (sucrose, glucose and fructose, also exert signalling roles through cross-talk with hormone and ROS signalling pathways and by mediating cell division and PCD. At the same time, emerging data indicate that sugar-derived signalling systems, including trehalose-6 phosphate (T6P, sucrose non-fermenting related kinase-1 (SnRK and the target of rapamycin (TOR kinase complex also play important roles in regulating plant development through modulating nutrient and energy signalling and metabolic processes, especially under abiotic stresses where sugar availability is low. This review aims to evaluate recent progress of research on abiotic stress responses of reproductive organs focusing on roles of sugar metabolism and signalling and addressing the possible biochemical and molecular mechanism by which sugars regulate fruit and seed set under heat and drought.

  16. Characterization, Leaching, and Filtration Testing for Bismuth Phosphate Sludge (Group 1) and Bismuth Phosphate Saltcake (Group 2) Actual Waste Sample Composites

    International Nuclear Information System (INIS)

    Lumetta, Gregg J.; Buck, Edgar C.; Daniel, Richard C.; Draper, Kathryn; Edwards, Matthew K.; Fiskum, Sandra K.; Hallen, Richard T.; Jagoda, Lynette K.; Jenson, Evan D.; Kozelisky, Anne E.; MacFarlan, Paul J.; Peterson, Reid A.; Shimskey, Rick W.; Sinkov, Sergey I.; Snow, Lanee A.

    2009-01-01

    A testing program evaluating actual tank waste was developed in response to Task 4 from the M-12 External Flowsheet Review Team (EFRT) issue response plan.() The test program was subdivided into logical increments. The bulk water-insoluble solid wastes that are anticipated to be delivered to the Waste Treatment and Immobilization Plant (WTP) were identified according to type such that the actual waste testing could be targeted to the relevant categories. Eight broad waste groupings were defined. Samples available from the 222S archive were identified and obtained for testing. The actual waste-testing program included homogenizing the samples by group, characterizing the solids and aqueous phases, and performing parametric leaching tests. Two of the eight defined groups - bismuth phosphate sludge (Group 1) and bismuth phosphate saltcake (Group 2) - are the subjects of this report. The Group 1 waste was anticipated to be high in phosphorus and was implicitly assumed to be present as BiPO4 (however, results presented here indicate that the phosphate in Group 1 is actually present as amorphous iron(III) phosphate). The Group 2 waste was also anticipated to be high in phosphorus, but because of the relatively low bismuth content and higher aluminum content, it was anticipated that the Group 2 waste would contain a mixture of gibbsite, sodium phosphate, and aluminum phosphate. Thus, the focus of the Group 1 testing was on determining the behavior of P removal during caustic leaching, and the focus of the Group 2 testing was on the removal of both P and Al. The waste-type definition, archived sample conditions, homogenization activities, characterization (physical, chemical, radioisotope, and crystal habit), and caustic leaching behavior as functions of time, temperature, and hydroxide concentration are discussed in this report. Testing was conducted according to TP-RPP-WTP-467

  17. SUGAR-SWEETENED BEVERAGE, SUGAR INTAKE OF INDIVIDUALS AND THEIR BLOOD PRESSURE: INTERMAP STUDY

    Science.gov (United States)

    Brown, Ian J.; Stamler, Jeremiah; Van Horn, Linda; Robertson, Claire E.; Chan, Queenie; Dyer, Alan R.; Huang, Chiang-Ching; Rodriguez, Beatriz L.; Zhao, Liancheng; Daviglus, Martha L.; Ueshima, Hirotsugu; Elliott, Paul

    2011-01-01

    The obesity epidemic has focused attention on relationships of sugars and sugar-sweetened beverages (SSB) to cardiovascular risk factors. Here we report cross-sectional associations of SSB, diet beverages, sugars with blood pressure (BP) for UK and USA participants of the International Study of Macro/Micro-nutrients and Blood Pressure (INTERMAP). Data collected includes four 24-h dietary recalls, two 24-h urine collections, eight BP readings, questionnaire data for 2,696 people ages 40-59 from 10 USA/UK population samples. Associations of SSB, diet beverages, and sugars (fructose, glucose, sucrose) with BP were assessed by multiple linear regression. Sugar-sweetened beverage intake related directly to BP, P-values 0.005 to Sugar-sweetened beverage intake higher by 1 serving/day (355 ml/24-h) was associated with systolic/diastolic BP differences of +1.6/+0.8 mm Hg (both P sugar-sodium interactions: for individuals with above-median 24-h urinary sodium excretion, fructose intake higher by 2 SD (5.6 %kcal) was associated with systolic/diastolic BP differences of +3.4/+2.2 mm Hg (both P sugar-BP differences for persons with higher sodium excretion, lend support to recommendations that intake of SSB, sugars, and salt be substantially reduced. PMID:21357284

  18. Enhancement of soil suppressiveness against Rhizoctonia solani in sugar beet by organic amendments

    NARCIS (Netherlands)

    Postma, J.; Schilder, M.T.

    2015-01-01

    The efficacy of different organic soil amendments on disease suppression to Rhizoctoniasolani AG 2-2IIIB was tested in a bio-assay with sugar beet as a test plant. Lysobacter populations in soil were quantified as a possible mechanism for disease suppression. Disease spread through the bio-assay

  19. Elucidation of the regulatory role of the fructose operon reveals a novel target for enhancing the NADPH supply in Corynebacterium glutamicum.

    Science.gov (United States)

    Wang, Zhihao; Chan, Siu Hung Joshua; Sudarsan, Suresh; Blank, Lars M; Jensen, Peter Ruhdal; Solem, Christian

    2016-11-01

    The performance of Corynebacterium glutamicum cell factories producing compounds which rely heavily on NADPH has been reported to depend on the sugar being metabolized. While some aspects of this phenomenon have been elucidated, there are still many unresolved questions as to how sugar metabolism is linked to redox and to the general metabolism. We here provide new insights into the regulation of the metabolism of this important platform organism by systematically characterizing mutants carrying various lesions in the fructose operon. Initially, we found that a strain where the dedicated fructose uptake system had been inactivated (KO-ptsF) was hampered in growth on sucrose minimal medium, and suppressor mutants appeared readily. Comparative genomic analysis in conjunction with enzymatic assays revealed that suppression was linked to inactivation of the pfkB gene, encoding a fructose-1-phosphate kinase. Detailed characterization of KO-ptsF, KO-pfkB and double knock-out (DKO) derivatives revealed a strong role for sugar-phosphates, especially fructose-1-phosphate (F1P), in governing sugar as well as redox metabolism due to effects on transcriptional regulation of key genes. These findings allowed us to propose a simple model explaining the correlation between sugar phosphate concentration, gene expression and ultimately the observed phenotype. To guide us in our analysis and help us identify bottlenecks in metabolism we debugged an existing genome-scale model onto which we overlaid the transcriptome data. Based on the results obtained we managed to enhance the NADPH supply and transform the wild-type strain into delivering the highest yield of lysine ever obtained on sucrose and fructose, thus providing a good example of how regulatory mechanisms can be harnessed for bioproduction. Copyright © 2016 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  20. Sucrose dependent mineral phosphate solubilization in Enterobacter asburiae PSI3 by heterologous overexpression of periplasmic invertases.

    Science.gov (United States)

    Kumar, Chanchal; Wagh, Jitendra; Archana, G; Naresh Kumar, G

    2016-12-01

    Enterobacter asburiae PSI3 solubilizes mineral phosphates in the presence of glucose by the secretion of gluconic acid generated by the action of a periplasmic pyrroloquinoline quinone dependent glucose dehydrogenase. In order to achieve mineral phosphate solubilization phenotype in the presence of sucrose, plasmids pCNK4 and pCNK5 containing genes encoding the invertase enzyme of Zymomonas mobilis (invB) and of Saccharomyces cerevisiae (suc2) under constitutive promoters were constructed with malE signal sequence (in case of invB alone as the suc2 is secreted natively). When introduced into E. asburiae PSI3, E. a. (pCNK4) and E. a. (pCNK5) transformants secreted 21.65 ± 0.94 and 22 ± 1.3 mM gluconic acid, respectively, in the presence of 75 mM sucrose and they also solubilized 180 ± 4.3 and 438 ± 7.3 µM P from the rock phosphate. In the presence of a mixture of 50 mM sucrose and 25 mM glucose, E. a. (pCNK5) secreted 34 ± 2.3 mM gluconic acid and released 479 ± 8.1 µM P. Moreover, in the presence of a mixture of eight sugars (10 mM each) in the medium, E. a. (pCNK5) released 414 ± 5.3 µM P in the buffered medium. Thus, this study demonstrates incorporation of periplasmic invertase imparted P solubilization ability to E. asburiae PSI3 in the presence of sucrose and mixture of sugars.

  1. Hypoxia-inducible factor-1 plays a role in phosphate-induced vascular smooth muscle cell calcification.

    Science.gov (United States)

    Mokas, Sophie; Larivière, Richard; Lamalice, Laurent; Gobeil, Stéphane; Cornfield, David N; Agharazii, Mohsen; Richard, Darren E

    2016-09-01

    Medial vascular calcification is a common complication of chronic kidney disease (CKD). Although elevated inorganic phosphate stimulates vascular smooth muscle cell (VSMC) osteogenic transdifferentiation and calcification, the mechanisms involved in their calcification during CKD are not fully defined. Because hypoxic gene activation is linked to CKD and stimulates bone cell osteogenic differentiation, we used in vivo and in vitro rodent models to define the role of hypoxic signaling during elevated inorganic phosphate-induced VSMC calcification. Cell mineralization studies showed that elevated inorganic phosphate rapidly induced VSMC calcification. Hypoxia strongly enhanced elevated inorganic phosphate-induced VSMC calcification and osteogenic transdifferentiation, as seen by osteogenic marker expression. Hypoxia-inducible factor-1 (HIF-1), the key hypoxic transcription factor, was essential for enhanced VSMC calcification. Targeting HIF-1 expression in murine VSMC blocked calcification in hypoxia with elevated inorganic phosphate while HIF-1 activators, including clinically used FG-4592/Roxadustat, recreated a procalcifying environment. Elevated inorganic phosphate rapidly activated HIF-1, even in normal oxygenation; an effect mediated by HIF-1α subunit stabilization. Thus, hypoxia synergizes with elevated inorganic phosphate to enhance VSMC osteogenic transdifferentiation. Our work identifies HIF-1 as an early CKD-related pathological event, prospective marker, and potential target against vascular calcification in CKD-relevant conditions. Copyright © 2016 International Society of Nephrology. Published by Elsevier Inc. All rights reserved.

  2. Physiological role of glucose-6-phosphate dehydrogenase in cold acclimation of strawberry (Fragaria × ananassa)

    Science.gov (United States)

    Zhang, Yong; Yu, Dingqun; Luo, Ya; Wang, Xiaorong; Chen, Qing; Sun, Bo; Wang, Yan; Liu, Zejing; Tang, Haoru

    2018-04-01

    In recent years, there has been an increasing interest in study of new resistance mechanism in fruit trees. All these regard the climate change and subsequent fruit production. Glucose-6-phosphate dehydrogenase (G6PDH) catalyzes the first and rate-limiting step of the oxidative pentose phosphate pathway (OPPP), and the expression of this enzyme is related to different biotic and abiotic stresses. Under accumulation of low temperature stress, the significant increase in G6PDH activity was found to be closely correlated to the levels of antioxidant enzymes, malondialdehyde (MDA) contents, sugar contents as well as changes of superoxide (O2•-). It is suggested that the enhancement of cold resistance of strawberry, which induced by cold acclimation, related to the significant increase in G6PDH activity. On one hand, G6PDH activates NADPH oxidase to generate reactive oxygen species (ROS); on the other hand, it may be involved in the activation of antioxidant enzymes, and accelerates many other important NADPH-dependent enzymatic reactions. Then further result in the elevation of membrane stability and cold resistance of strawberry. Interestingly, even though the plants were placed again under a temperature of 25°C for 1 d, the higher cold resistance, enzyme activities and soluble sugar content acquired.

  3. Rock phosphate solubilizing and cellulolytic actinomycete isolates of earthworm casts

    Science.gov (United States)

    Mba, Caroline C.

    1994-03-01

    Four microbial isolates, OP2, OP3, OP6, and OP7, of earthworm casts of Pontoscolex corethrurus were found to be acid tolerant actinomycetes and efficient rock phosphate (RP) solubilizers that could grow fast on NH4Cl-enriched or N-free carboxymethyl cellulose or glucose as sole carbon source. CMC (carboxymethyl cellulose) induced production of extracellular cellulase enzyme and the production of reducing sugar in all the isolates. RP solubilizing power was observed to be inversely related to glucose consumption. The most efficient RP solubilizer was found to consume the least glucose. Growth was faster on cellulose than on glucose media. N-free CMC induced greater glucose production than NH4Cl-enriched CMC medium. Both CMC and glucose media were acidified by all the isolates, however, RP solubilizing power decreased with acidification. Solubilization power was greatest with isolate OP7, which also produced the greatest amount of reducing sugar per gram CMC. Both RP solubilizing power and the cellulolytic efficiency varied among isolates. A minimum of 631 µg P/0.1 g RP and a maximum of 951.4 µg P/0.1 g RP was recorded.

  4. Fructose 1-phosphate is the preferred effector of the metabolic regulator Cra of Pseudomonas putida.

    Science.gov (United States)

    Chavarría, Max; Santiago, César; Platero, Raúl; Krell, Tino; Casasnovas, José M; de Lorenzo, Víctor

    2011-03-18

    The catabolite repressor/activator (Cra) protein is a global sensor and regulator of carbon fluxes through the central metabolic pathways of gram-negative bacteria. To examine the nature of the effector (or effectors) that signal such fluxes to the protein of Pseudomonas putida, the Cra factor of this soil microorganism has been purified and characterized and its three-dimensional structure determined. Analytical ultracentrifugation, gel filtration, and mobility shift assays showed that the effector-free Cra is a dimer that binds an operator DNA sequence in the promoter region of the fruBKA cluster. Furthermore, fructose 1-phosphate (F1P) was found to most efficiently dissociate the Cra-DNA complex. Thermodynamic parameters of the F1P-Cra-DNA interaction calculated by isothermal titration calorimetry revealed that the factor associates tightly to the DNA sequence 5'-TTAAACGTTTCA-3' (K(D) = 26.3 ± 3.1 nM) and that F1P binds the protein with an apparent stoichiometry of 1.06 ± 0.06 molecules per Cra monomer and a K(D) of 209 ± 20 nM. Other possible effectors, like fructose 1,6-bisphosphate, did not display a significant affinity for the regulator under the assay conditions. Moreover, the structure of Cra and its co-crystal with F1P at a 2-Å resolution revealed that F1P fits optimally the geometry of the effector pocket. Our results thus single out F1P as the preferred metabolic effector of the Cra protein of P. putida.

  5. Fructose 1-Phosphate Is the Preferred Effector of the Metabolic Regulator Cra of Pseudomonas putida*

    Science.gov (United States)

    Chavarría, Max; Santiago, César; Platero, Raúl; Krell, Tino; Casasnovas, José M.; de Lorenzo, Víctor

    2011-01-01

    The catabolite repressor/activator (Cra) protein is a global sensor and regulator of carbon fluxes through the central metabolic pathways of Gram-negative bacteria. To examine the nature of the effector (or effectors) that signal such fluxes to the protein of Pseudomonas putida, the Cra factor of this soil microorganism has been purified and characterized and its three-dimensional structure determined. Analytical ultracentrifugation, gel filtration, and mobility shift assays showed that the effector-free Cra is a dimer that binds an operator DNA sequence in the promoter region of the fruBKA cluster. Furthermore, fructose 1-phosphate (F1P) was found to most efficiently dissociate the Cra-DNA complex. Thermodynamic parameters of the F1P-Cra-DNA interaction calculated by isothermal titration calorimetry revealed that the factor associates tightly to the DNA sequence 5′-TTAAACGTTTCA-3′ (KD = 26.3 ± 3.1 nm) and that F1P binds the protein with an apparent stoichiometry of 1.06 ± 0.06 molecules per Cra monomer and a KD of 209 ± 20 nm. Other possible effectors, like fructose 1,6-bisphosphate, did not display a significant affinity for the regulator under the assay conditions. Moreover, the structure of Cra and its co-crystal with F1P at a 2-Å resolution revealed that F1P fits optimally the geometry of the effector pocket. Our results thus single out F1P as the preferred metabolic effector of the Cra protein of P. putida. PMID:21239488

  6. Development of real-time PCR method for the detection and the quantification of a new endogenous reference gene in sugar beet "Beta vulgaris L.": GMO application.

    Science.gov (United States)

    Chaouachi, Maher; Alaya, Akram; Ali, Imen Ben Haj; Hafsa, Ahmed Ben; Nabi, Nesrine; Bérard, Aurélie; Romaniuk, Marcel; Skhiri, Fethia; Saïd, Khaled

    2013-01-01

    KEY MESSAGE : Here, we describe a new developed quantitative real-time PCR method for the detection and quantification of a new specific endogenous reference gene used in GMO analysis. The key requirement of this study was the identification of a new reference gene used for the differentiation of the four genomic sections of the sugar beet (Beta vulgaris L.) (Beta, Corrollinae, Nanae and Procumbentes) suitable for quantification of genetically modified sugar beet. A specific qualitative polymerase chain reaction (PCR) assay was designed to detect the sugar beet amplifying a region of the adenylate transporter (ant) gene only from the species of the genomic section I of the genus Beta (cultivated and wild relatives) and showing negative PCR results for 7 species of the 3 other sections, 8 related species and 20 non-sugar beet plants. The sensitivity of the assay was 15 haploid genome copies (HGC). A quantitative real-time polymerase chain reaction (QRT-PCR) assay was also performed, having high linearity (R (2) > 0.994) over sugar beet standard concentrations ranging from 20,000 to 10 HGC of the sugar beet DNA per PCR. The QRT-PCR assay described in this study was specific and more sensitive for sugar beet quantification compared to the validated test previously reported in the European Reference Laboratory. This assay is suitable for GMO quantification in routine analysis from a wide variety of matrices.

  7. Competitive binding assay for fructose 2,6-bisphosphate

    International Nuclear Information System (INIS)

    Thomas, H.; Uyeda, K.

    1986-01-01

    A new direct assay method for fructose 2,6-bisphosphate has been developed based on competitive binding of labeled and unlabeled fructose 2,6-P 2 to phosphofructokinase. Phosphofructokinase (0.5-1.3 pmol promoter) is incubated with saturating concentrations (5.0-5.5 pmol) of fructose 2,6[2- 32 P]P 2 and samples containing varying concentrations of fructose 2,6-P 2 . The resulting stable binary complex is retained on nitrocellulose filters with a binding efficiency of up to 70%. Standard curves obtained with this assay show strict linearity with varying fructose 2,6-P 2 in the range of 0.5 to 45 pmol, which exceeds the sensitivity of most of the previously described assay methods. Fructose 2,6-P 2 , ATP, and high concentrations of phosphate interfere with this assay. However, the extent of this inhibition is negligible since their tissue contents are one-half to one-tenth that examined. The new assay is simple, direct, rapid, and does not require pretreatment

  8. The role of sphingosine-1-phosphate (S1P) and ceramide-1-phosphate (C1P) in the trafficking of normal and malignant cells

    Science.gov (United States)

    Ratajczak, Mariusz Z.; Suszynska, Malwina; Borkowska, Sylwia; Ratajczak, Janina; Schneider, Gabriela

    2014-01-01

    Introduction A common feature of many types of cells is their responsiveness to chemotactic gradients of factors for which they express the corresponding receptors. The most studied chemoattractants so far are peptide-based growth factors and a family of cytokines endowed with strong chemotactic properties, called chemokines. However, additional evidence has accumulated that, in addition to these peptide-based chemoattractants, an important role in cell migration is played by bioactive lipids. Areas covered Solid evidence has accumulated that two bioactive phosphorylated sphingolipids that are derivatives of sphingolipid metabolism, namely sphingosine-1-phosphate (S1P) and ceramide-1-phosphate (C1P), are potent chemoattractants for a variety of cells. In this review, we will discuss the effect of these two phosphorylated sphingolipids on the trafficking of normal and malignant cells, and, in particular, we will focus on their role in trafficking of normal hematopoietic stem/progenitor cells. Unlike other mediators, S1P under steady state conditions maintain a steep gradient between interstitial fluid and peripheral blood and lymph across the endothelial barrier, which is important in the egress of cells from bone marrow. Both S1P and C1P may be upregulated in damaged tissues, which may result in reversal of this gradient. Expert opinion S1P and C1P are important regulators of the trafficking of normal and malignant cells, and modification of their biological effects will have important applications in optimizing stem cell mobilization and homing, tissue organ/regeneration, and preventing cancer metastasis. PMID:24188167

  9. Fructose 1-phosphate is the one and only physiological effector of the Cra (FruR) regulator of Pseudomonas putida.

    Science.gov (United States)

    Chavarría, Max; Durante-Rodríguez, Gonzalo; Krell, Tino; Santiago, César; Brezovsky, Jan; Damborsky, Jiri; de Lorenzo, Víctor

    2014-01-01

    Fructose-1-phosphate (F1P) is the preferred effector of the catabolite repressor/activator (Cra) protein of the soil bacterium Pseudomonas putida but its ability to bind other metabolic intermediates in vivo is unclear. The Cra protein of this microorganism (Cra(PP)) was submitted to mobility shift assays with target DNA sequences (the PfruB promoter) and candidate effectors fructose-1,6-bisphosphate (FBP), glucose 6-phosphate (G6P), and fructose-6-phosphate (F6P). 1 mM F1P was sufficient to release most of the Cra protein from its operators but more than 10 mM of FBP or G6P was required to free the same complex. However, isothermal titration microcalorimetry failed to expose any specific interaction between Cra(PP) and FBP or G6P. To solve this paradox, transcriptional activity of a PfruB-lacZ fusion was measured in wild-type and ΔfruB cells growing on substrates that change the intracellular concentrations of F1P and FBP. The data indicated that PfruB activity was stimulated by fructose but not by glucose or succinate. This suggested that Cra(PP) represses expression in vivo of the cognate fruBKA operon in a fashion dependent just on F1P, ruling out any other physiological effector. Molecular docking and dynamic simulations of the Cra-agonist interaction indicated that both metabolites can bind the repressor, but the breach in the relative affinity of Cra(PP) for F1P vs FBP is three orders of magnitude larger than the equivalent distance in the Escherichia coli protein. This assigns the Cra protein of P. putida the sole role of transducing the presence of fructose in the medium into a variety of direct and indirect physiological responses.

  10. A new strategy for the cloning, overexpression and one step purification of three DHAP-dependent aldolases: rhamnulose-1-phosphate aldolase, fuculose-1-phosphate aldolase and tagatose-1,6-diphosphate aldolase.

    Science.gov (United States)

    Garcia-Junceda, E; Shen, G J; Sugai, T; Wong, C H

    1995-07-01

    Three DHAP-dependent aldolases, rhamnulose-1-phosphate aldolase (Rham-1PA), fuculose-1-phosphate aldolase (Fuc-1PA) and tagatose-1,6-diphosphate aldolase (TDPA) have been cloned and overexpressed in Escherichia coli using two different expression vectors: pTrcHis for the expression of Rham-1PA and Fuc-1PA and pRSET for the expression of TDPA. In each case the recombinant enzyme is synthesized as a fusion protein with a hexahistidine tag on the N-terminus. The three enzymes have been purified in only one step by chelation affinity chromatography. The effects of cultivation temperature and concentration of inducer have been studied in order to optimize the expression of the recombinant proteins and to avoid the formation of inclusion bodies.

  11. Sugar-induced cephalic-phase insulin release is mediated by a T1r2+T1r3-independent taste transduction pathway in mice.

    Science.gov (United States)

    Glendinning, John I; Stano, Sarah; Holter, Marlena; Azenkot, Tali; Goldman, Olivia; Margolskee, Robert F; Vasselli, Joseph R; Sclafani, Anthony

    2015-09-01

    Sensory stimulation from foods elicits cephalic phase responses, which facilitate digestion and nutrient assimilation. One such response, cephalic-phase insulin release (CPIR), enhances glucose tolerance. Little is known about the chemosensory mechanisms that activate CPIR. We studied the contribution of the sweet taste receptor (T1r2+T1r3) to sugar-induced CPIR in C57BL/6 (B6) and T1r3 knockout (KO) mice. First, we measured insulin release and glucose tolerance following oral (i.e., normal ingestion) or intragastric (IG) administration of 2.8 M glucose. Both groups of mice exhibited a CPIR following oral but not IG administration, and this CPIR improved glucose tolerance. Second, we examined the specificity of CPIR. Both mouse groups exhibited a CPIR following oral administration of 1 M glucose and 1 M sucrose but not 1 M fructose or water alone. Third, we studied behavioral attraction to the same three sugar solutions in short-term acceptability tests. B6 mice licked more avidly for the sugar solutions than for water, whereas T1r3 KO mice licked no more for the sugar solutions than for water. Finally, we examined chorda tympani (CT) nerve responses to each of the sugars. Both mouse groups exhibited CT nerve responses to the sugars, although those of B6 mice were stronger. We propose that mice possess two taste transduction pathways for sugars. One mediates behavioral attraction to sugars and requires an intact T1r2+T1r3. The other mediates CPIR but does not require an intact T1r2+T1r3. If the latter taste transduction pathway exists in humans, it should provide opportunities for the development of new treatments for controlling blood sugar. Copyright © 2015 the American Physiological Society.

  12. GATA4-mediated cardiac hypertrophy induced by D-myo-inositol 1,4,5-tris-phosphate

    International Nuclear Information System (INIS)

    Zhu Zhiming; Zhu Shanjun; Liu Daoyan; Yu Zengping; Yang Yongjian; Giet, Markus van der; Tepel, Martin

    2005-01-01

    We evaluated the effects of D-myo-inositol 1,4,5-tris-phosphate on cardiac hypertrophy. D-myo-inositol 1,4,5-tris-phosphate augmented cardiac hypertrophy as evidenced by its effects on DNA synthesis, protein synthesis, and expression of immediate-early genes c-myc and c-fos, β-myosin heavy chain, and α-actin. The administration of D-myo-inositol 1,4,5-tris-phosphate increased the expression of nuclear factor of activated T-cells and cardiac-restricted zinc finger transcription factor (GATA4). Real-time quantitative RT-PCR showed that D-myo-inositol 1,4,5-tris-phosphate-induced GATA4 mRNA was significantly enhanced even in the presence of the calcineurin inhibitor, cyclosporine A. The effect of D-myo-inositol 1,4,5-tris-phosphate was blocked after inhibition of inositol-trisphosphate receptors but not after inhibition of c-Raf/mitogen-activated protein kinase kinase (MEK)/mitogen-activated protein kinase (ERK) or p38 mitogen-activated protein kinase pathways. The study shows that D-myo-inositol 1,4,5-tris-phosphate-induced cardiac hypertrophy is mediated by GATA4 but independent from the calcineurin pathway

  13. Hierarchy of non-glucose sugars in Escherichia coli.

    Science.gov (United States)

    Aidelberg, Guy; Towbin, Benjamin D; Rothschild, Daphna; Dekel, Erez; Bren, Anat; Alon, Uri

    2014-12-24

    Understanding how cells make decisions, and why they make the decisions they make, is of fundamental interest in systems biology. To address this, we study the decisions made by E. coli on which genes to express when presented with two different sugars. It is well-known that glucose, E. coli's preferred carbon source, represses the uptake of other sugars by means of global and gene-specific mechanisms. However, less is known about the utilization of glucose-free sugar mixtures which are found in the natural environment of E. coli and in biotechnology. Here, we combine experiment and theory to map the choices of E. coli among 6 different non-glucose carbon sources. We used robotic assays and fluorescence reporter strains to make precise measurements of promoter activity and growth rate in all pairs of these sugars. We find that the sugars can be ranked in a hierarchy: in a mixture of a higher and a lower sugar, the lower sugar system shows reduced promoter activity. The hierarchy corresponds to the growth rate supported by each sugar- the faster the growth rate, the higher the sugar on the hierarchy. The hierarchy is 'soft' in the sense that the lower sugar promoters are not completely repressed. Measurement of the activity of the master regulator CRP-cAMP shows that the hierarchy can be quantitatively explained based on differential activation of the promoters by CRP-cAMP. Comparing sugar system activation as a function of time in sugar pair mixtures at sub-saturating concentrations, we find cases of sequential activation, and also cases of simultaneous expression of both systems. Such simultaneous expression is not predicted by simple models of growth rate optimization, which predict only sequential activation. We extend these models by suggesting multi-objective optimization for both growing rapidly now and preparing the cell for future growth on the poorer sugar. We find a defined hierarchy of sugar utilization, which can be quantitatively explained by

  14. Sugars and Desiccation Tolerance in Seeds 1

    Science.gov (United States)

    Koster, Karen L.; Leopold, A. Carl

    1988-01-01

    Soluble sugars have been shown to protect liposomes and lobster microsomes from desiccation damage, and a protective role has been proposed for them in several anhydrous systems. We have studied the relationship between soluble sugar content and the loss of desiccation tolerance in the axes of germinating soybean (Glycine max L. Merr. cv Williams), pea (Pisum sativum L. cv Alaska), and corn (Zea mays L. cv Merit) axes. The loss of desiccation tolerance during imbibition was monitored by following the ability of seeds to germinate after desiccation following various periods of preimbibition and by following the rates of electrolyte leakage from dried, then rehydrated axes. Finally, we analyzed the soluble sugar contents of the axes throughout the transition from desiccation tolerance to intolerance. These analyses show that sucrose and larger oligosaccharides were consistently present during the tolerant stage, and that desiccation tolerance disappeared as the oligosaccharides were lost. The results support the idea that sucrose may serve as the principal agent of desiccation tolerance in these seeds, with the larger oligosaccharides serving to keep the sucrose from crystallizing. PMID:16666392

  15. Agonist-dependent effects of mutations in the sphingosine-1-phosphate type 1 receptor

    NARCIS (Netherlands)

    van Loenen, Pieter B.; de Graaf, Chris; Verzijl, Dennis; Leurs, Rob; Rognan, Didier; Peters, Stephan L. M.; Alewijnse, Astrid E.

    2011-01-01

    The sphingosine-1-phosphate type 1 (S1P(1)) receptor is a new target in the treatment of auto-immune diseases as evidenced by the recent approval of FTY720 (Fingolimod). The ligand-binding pocket of the S1P(1) receptor has been generally characterised but detailed insight into ligand-specific

  16. sphingosine-1-phosphate transport and its role in immunology

    NARCIS (Netherlands)

    Reitsema, V.; Bouma, Hjalmar; Kok, Jan

    2014-01-01

    Sphingosine-1-phosphate (S1P) is a sphingolipid metabolite with many important functions in cellular and systemic physiology, including the immune system. As it cannot traverse the membrane, it is exported from cells by transporters. Several members of the ATP-binding cassette (ABC) transporter

  17. Sugar-binding sites of the HA1 subcomponent of Clostridium botulinum type C progenitor toxin.

    Science.gov (United States)

    Nakamura, Toshio; Tonozuka, Takashi; Ide, Azusa; Yuzawa, Takayuki; Oguma, Keiji; Nishikawa, Atsushi

    2008-02-22

    Clostridium botulinum type C 16S progenitor toxin contains a hemagglutinin (HA) subcomponent, designated HA1, which appears to play an important role in the effective internalization of the toxin in gastrointestinal epithelial cells and in creating a broad specificity for the oligosaccharide structure that corresponds to various targets. In this study, using the recombinant protein fused to glutathione S-transferase, we investigated the binding specificity of the HA1 subcomponent to sugars and estimated the binding sites of HA1 based on X-ray crystallography and soaking experiments using various sugars. N-Acetylneuraminic acid, N-acetylgalactosamine, and galactose effectively inhibited the binding that occurs between glutathione S-transferase-HA1 and mucins, whereas N-acetylglucosamine and glucose did not inhibit it. The crystal structures of HA1 complex with N-acetylneuraminic acid, N-acetylgalactosamine, and galactose were also determined. There are two sugar-binding sites, sites I and II. Site I corresponds to the electron densities noted for all sugars and is located at the C-terminal beta-trefoil domain, while site II corresponds to the electron densities noted only for galactose. An aromatic amino acid residue, Trp176, at site I has a stacking interaction with the hexose ring of the sugars. On the other hand, there is no aromatic residue at site II; thus, the interaction with galactose seems to be poor. The double mutant W176A at site I and D271F at site II has no avidity for N-acetylneuraminic acid but has avidity for galactose. In this report, the binding specificity of botulinum C16S toxin HA1 to various sugars is demonstrated based on its structural features.

  18. Combined subcritical water and enzymatic hydrolysis for reducing sugar production from coconut husk

    Science.gov (United States)

    Muharja, Maktum; Junianti, Fitri; Nurtono, Tantular; Widjaja, Arief

    2017-05-01

    Coconut husk wastes are abundantly available in Indonesia. It has a potential to be used into alternative renewable energy sources such as hydrogen using enzymatic hydrolysis followed by a fermentation process. Unfortunately, enzymatic hydrolysis is hampered by the complex structure of lignocellulose, so the cellulose component is hard to degrade. In this study, Combined Subcritical Water (SCW) and enzymatic hydrolysis are applied to enhance fermentable, thereby reducing production of sugar from coconut husk. There were two steps in this study, the first step was coconut husk pretreated by SCW in batch reactor at 80 bar and 150-200°C for 60 minutes reaction time. Secondly, solid fraction from the results of SCW was hydrolyzed using the mixture of pure cellulose and xylanase enzymes. Analysis was conducted on untreated and SCW-treated by gravimetric assay, liquid fraction after SCW and solid fraction after enzymatic hydrolysis using DNS assay. The maximum yield of reducing sugar (including xylose, arabinose glucose, galactose, mannose) was 1.254 gr per 6 gr raw material, representing 53.95% of total sugar in coconut husk biomass which was obtained at 150°C 80 bar for 60 minutes reaction time of SCW-treated and 6 hour of enzymatic hydrolysis using mixture of pure cellulose and xylanase enzymes (18.6 U /gram of coconut husk).

  19. Alpha-tocopheryl phosphate: a novel, natural form of vitamin E.

    Science.gov (United States)

    Gianello, Robert; Libinaki, Roksan; Azzi, Angelo; Gavin, Paul D; Negis, Yesim; Zingg, Jean-Marc; Holt, Phillip; Keah, Hooi-Hong; Griffey, Annike; Smallridge, Andrew; West, Simon M; Ogru, Esra

    2005-10-01

    We have detected alpha-tocopheryl phosphate in biological tissues including liver and adipose tissue, as well as in a variety of foods, suggesting a ubiquitous presence in animal and plant tissue. Alpha-tocopheryl phosphate is a water-soluble molecule that is resistant to both acid and alkaline hydrolysis, making it undetectable using standard assays for vitamin E. A new method was therefore developed to allow the extraction of both alpha-tocopheryl phosphate and alpha-tocopherol from a single specimen. We used ESMS to detect endogenous alpha-tocopheryl phosphate in biological samples that also contained alpha-tocopherol. Due to the significance of these findings, further proof was required to unequivocally demonstrate the presence of endogenous alpha-tocopheryl phosphate in biological samples. Four independent methods of analysis were examined: HPLC, LCMS, LCMS/MS, and GCMS. Alpha-tocopherol phosphate was identified in all instances by comparison between standard alpha-tocopheryl phosphate and extracts of biological tissues. The results show that alpha-tocopheryl phosphate is a natural form of vitamin E. The discovery of endogenous alpha-tocopheryl phosphate has implications for the expanding knowledge of the roles of alpha-tocopherol in biological systems.

  20. Patterns of sugar feeding and host plant preferences in adult males of An. gambiae (Diptera: Culicidae).

    Science.gov (United States)

    Gouagna, Louis-Clément; Poueme, Rodrigue S; Dabiré, Kounbobr Roch; Ouédraogo, Jean-Bosco; Fontenille, Didier; Simard, Frédéric

    2010-12-01

    Sugar feeding by male mosquitoes is critical for their success in mating competition. However, the facets of sugar source finding under natural conditions remain unknown. Here, evidence obtained in Western Burkina Faso indicated that the distribution of An. gambiae s.s. (M and S molecular forms) males across different peri-domestic habitats is dependent on the availability of potential sugar sources from which they obtain more favorable sites for feeding or resting. Among field-collected anophelines, a higher proportion of specimens containing fructose were found on flowering Mangifera indica (Anacardiaceae), Dolonix regia (Fabaceae), Thevetia neriifolia (Apocynaceae), Senna siamea, and Cassia sieberiana (both Fabaceae) compared to that recorded on other nearby plants, suggesting that some plants are favored for use as a sugar source over others. Y-tube olfactometer assays with newly-emerged An. gambiae s.s. exposed to odors from individual plants and some combinations thereof showed that males use odor cues to guide their preference. The number of sugar-positive males was variable in a no-choice cage assay, consistent with the olfactory response patterns towards corresponding odor stimuli. These experiments provide the first evidence both in field and laboratory conditions for previously unstudied interactions between males of An. gambiae and natural sugar sources. © 2010 The Society for Vector Ecology.

  1. The sphingosine 1-phosphate receptor S1P(2) triggers hepatic wound healing

    NARCIS (Netherlands)

    Serriere-Lanneau, Valerie; Teixeira-Clerc, Fatima; Li, Liying; Schippers, Marlies; de Wries, Willie; Julien, Boris; Tran-Van-Nhieu, Jeanne; Manin, Sylvie; Poelstra, Klaas; Chun, Jerold; Carpentier, Stephane; Levade, Thierry; Mallat, Ariane; Lotersztajn, Sophie

    Sphingosine 1-phosphate (S1P) is a bioactive sphingolipid produced by sphingosine kinase (SphK1 and 2). We previously showed that S1P receptors (S1P(1), S1P(2), and S1P(3)) are expressed in hepatic myofibroblasts (hMF), a population of cells that triggers matrix remodeling during liver injury. Here

  2. An alpha-glucose-1-phosphate phosphodiesterase is present in rat liver cytosol

    International Nuclear Information System (INIS)

    Srisomsap, C.; Richardson, K.L.; Jay, J.C.; Marchase, R.B.

    1989-01-01

    UDP-glucose:glycoprotein glucose-1-phosphotransferase (Glc-phosphotransferase) catalyzes the transfer of alpha-Glc-1-P from UDP-Glc to mannose residues on acceptor glycoproteins. The predominant acceptor for this transfer in both mammalian cells and Paramecium is a cytoplasmic glycoprotein of 62-63 kDa. When cytoplasmic proteins from rat liver were fractionated by preparative isoelectric focusing following incubation of a liver homogenate with the 35S-labeled phosphorothioate analogue of UDP-Glc ([beta-35S]UDP-Glc), the acceptor was found to have a pI of about 6.0. This fraction, when not labeled prior to the focusing, became very heavily labeled when mixed with [beta-35S]. UDP-Glc and intact liver microsomes, a rich source of the Glc-phosphotransferase. In addition, it was observed that the isoelectric fractions of the cytosol having pI values of 2-3.2 contained a degradative activity, alpha-Glc-1-P phosphodiesterase, that was capable of removing alpha-Glc-1-P, monitored through radioactive labeling both in the sugar and the phosphate, as an intact unit from the 62-kDa acceptor. Identification of the product of this cleavage was substantiated by its partial transformation to UDP-Glc in the presence of UTP and UDP-Glc pyrophosphorylase. The alpha-Glc-1-P phosphodiesterase had a pH optimum of 7.5 and was not effectively inhibited by any of the potential biochemical inhibitors that were tested. Specificity for the Glc-alpha-1-P-6-Man diester was suggested by the diesterase's inability to degrade UDP-Glc or glucosylphosphoryldolichol. This enzyme may be important in the regulation of secretion since the alpha-Glc-1-P present on the 62-kDa phosphoglycoprotein appears to be removed and then rapidly replaced in response to secretagogue

  3. Formulation of single super phosphate fertilizer from rock phosphate of Hazara, Pakistan

    Directory of Open Access Journals (Sweden)

    Matiullah Khan

    2012-05-01

    Full Text Available Phosphorus deficiency is wide spread in soils of Pakistan. It is imperative to explore the potential and economics of indigenous Hazara rock phosphate for preparation of single super phosphate fertilizer. For the subject study rock phosphate was collected from Hazara area ground at 160 mesh level with 26% total P2O5 content for manual preparation of single super phosphate fertilizer. The rock phosphate was treated with various concentrations of sulfuric acid (98.9%, diluted or pure in the field. The treatments comprised of 20 and 35% pure acid and diluted with acid-water ratios of 1:5, 1:2, 1:1 and 2:1 v/v for acidulation at the rate of 60 liters 100 kg-1 rock phosphate. The amount was prior calculated in the laboratory for complete wetting of rock phosphate. A quantity of 150 kg rock phosphate was taken as treatment. The respective amount of acid was applied with the spray pump of stainless steel or poured with bucket. After proper processing, chemical analysis of the products showed a range of available P2O5 content from 9.56 to 19.24% depending upon the amount of acid and its dilution. The results reveal at that 1:1 dilutions gave the highest P2O5 content (19.24%, lowest free acid (6 % and 32% weight increase. The application of acid beyond or below this combination either pure or diluted gave hygroscopic product and higher free acids. The cost incurred upon the manual processing was almost half the prevailing rates in the market. These results lead to conclude that application of sulfuric acid at the rate of 60 liters 100 kg-1 with the dilution of 50% (v/v can yield better kind of SSP from Hazara rock phosphate at lower prices.

  4. Measurement of 2-carboxyarabinitol 1-phosphate in plant leaves by isotope dilution

    International Nuclear Information System (INIS)

    Moore, B.D.; Kobza, J.; Seemann, J.R.

    1991-01-01

    The level of 2-carboxyarabinitol 1-phosphate (CA1P) in leaves of 12 species was determined by an isotope dilution assay. 14 C-labeled standard was synthesized from [2- 14 C]carboxyarabinitol 1,5-bisphosphate using acid phosphatase, and was added at the initial point of leaf extraction. Leaf CA1P was purified and its specific activity determined. CA1P was found in dark-treated leaves of all species examined, including spinach (Spinacea oleracea), wheat (Triticum aestivum), Arabidopsis thaliana, and maize (Zea mays). The highest amounts were found in bean (Phaseolus vulgaris) and petunia (Petunia hybrida), which had 1.5 to 1.8 moles CA1P per mole ribulose 1,5-bisphosphate carboxylase catalytic sites. Most species had intermediate amounts of CA1P (0.2 to 0.8 mole CA1P per mole catalytic sites). Such intermediate to high levels of CA1P support the hypothesis that CA1P functions in many species as a light-dependent regulator of ribulose 1,5-bisphosphate carboxylase activity and whole leaf photosynthetic CO 2 assimilation. However, CA1P levels in spinach, wheat, and A. thaliana were particularly low (less than 0.09 mole CA1P per mole catalytic sites). In such species, CA1P does not likely have a significant role in regulating ribulose 1,5-bisphosphate carboxylase activity, but could have a different physiological role

  5. Hawkmoths use nectar sugar to reduce oxidative damage from flight.

    Science.gov (United States)

    Levin, E; Lopez-Martinez, G; Fane, B; Davidowitz, G

    2017-02-17

    Nectar-feeding animals have among the highest recorded metabolic rates. High aerobic performance is linked to oxidative damage in muscles. Antioxidants in nectar are scarce to nonexistent. We propose that nectarivores use nectar sugar to mitigate the oxidative damage caused by the muscular demands of flight. We found that sugar-fed moths had lower oxidative damage to their flight muscle membranes than unfed moths. Using respirometry coupled with δ 13 C analyses, we showed that moths generate antioxidant potential by shunting nectar glucose to the pentose phosphate pathway (PPP), resulting in a reduction in oxidative damage to the flight muscles. We suggest that nectar feeding, the use of PPP, and intense exercise are causally linked and have allowed the evolution of powerful fliers that feed on nectar. Copyright © 2017, American Association for the Advancement of Science.

  6. The relationship between measures of tree vigor and pear thrips damage in sugar maple

    Science.gov (United States)

    Gretchen Smith; Christina M. Petersen; Roy Van Driesche; Charles Burnham

    1991-01-01

    In this presentation I will address three points associated with pear thrips damage and sugar maple. First, I will describe the impact of pear thrips on sugar maple in Massachusetts, in both the sugarbush and the natural forest stand, based on root starch assays that were completed this fall (1988). Secondly, I will discuss the relationship between tree health and...

  7. Electrophysiological and functional effects of sphingosine-1-phosphate in mouse ventricular fibroblasts

    Energy Technology Data Exchange (ETDEWEB)

    Benamer, Najate [UMR CNRS/Universite de Poitiers No. 6187, Pole Biologie Sante Bat B36, BP 633, 1 rue Georges Bonnet, 86022 Poitiers (France); Fares, Nassim [Laboratoire de Physiologie, Faculte de Medecine, Universite Saint Joseph, Beyrouth (Lebanon); Bois, Patrick [UMR CNRS/Universite de Poitiers No. 6187, Pole Biologie Sante Bat B36, BP 633, 1 rue Georges Bonnet, 86022 Poitiers (France); Faivre, Jean-Francois, E-mail: Jean-Francois.Faivre@univ-poitiers.fr [UMR CNRS/Universite de Poitiers No. 6187, Pole Biologie Sante Bat B36, BP 633, 1 rue Georges Bonnet, 86022 Poitiers (France)

    2011-04-29

    Highlights: {yields} In cardiac fibroblasts, SUR2/Kir6.1 channel is activated by S1P via the S1P3R. {yields} S1P increases cell proliferation through SUR2/Kir6.1 activation. {yields} S1P decreases collagen and IL-6 secretion through SUR2/Kir6.1 activation. {yields} S1P stimulates fibroblast migration independently from SUR2/Kir6.1 channel. -- Abstract: The aim of this study was to characterize the effects of sphingosine-1-phosphate (S1P) on cardiac ventricular fibroblasts. Impacts of S1P on fibroblast excitability, cell migration, proliferation and secretion were characterized. The patch-clamp technique in the whole-cell configuration was used to study the S1P-induced current from mouse ventricular fibroblasts. The expression level of the S1P receptor during cell culture duration was evaluated by western-blot. Fibroblast proliferation and migration were quantified using the methylene blue assay and the Boyden chamber technique, respectively. Finally, fibroblast secretion properties were estimated by quantification of the IL-6 and collagen levels using ELISA and SIRCOL collagen assays, respectively. We found that S1P activated SUR2/Kir6.1 channel and that this effect was sensitive to specific inhibition of the S1P receptor of type 3 (S1P3R). In contrast, S1P1R receptor inhibition had no effect. Moreover, the S1P-induced current increased with cell culture duration whereas S1P3R expression level remained constant. The activation of SUR2/Kir6.1 channel by S1P via S1P3R stimulated cell proliferation and decreased IL-6 and collagen secretions. S1P also stimulated fibroblast migration via S1P3R but independently from SUR2/Kir6.1 channel activation. This study demonstrates that S1P, via S1P3R, affects cardiac ventricular fibroblasts function independently or through activation of SUR2/Kir6.1 channel. The latter effect occurs after fibroblasts differentiate into myofibroblasts, opening a new potential therapeutic strategy to modulate fibrosis after cardiac

  8. Electrophysiological and functional effects of sphingosine-1-phosphate in mouse ventricular fibroblasts

    International Nuclear Information System (INIS)

    Benamer, Najate; Fares, Nassim; Bois, Patrick; Faivre, Jean-Francois

    2011-01-01

    Highlights: → In cardiac fibroblasts, SUR2/Kir6.1 channel is activated by S1P via the S1P3R. → S1P increases cell proliferation through SUR2/Kir6.1 activation. → S1P decreases collagen and IL-6 secretion through SUR2/Kir6.1 activation. → S1P stimulates fibroblast migration independently from SUR2/Kir6.1 channel. -- Abstract: The aim of this study was to characterize the effects of sphingosine-1-phosphate (S1P) on cardiac ventricular fibroblasts. Impacts of S1P on fibroblast excitability, cell migration, proliferation and secretion were characterized. The patch-clamp technique in the whole-cell configuration was used to study the S1P-induced current from mouse ventricular fibroblasts. The expression level of the S1P receptor during cell culture duration was evaluated by western-blot. Fibroblast proliferation and migration were quantified using the methylene blue assay and the Boyden chamber technique, respectively. Finally, fibroblast secretion properties were estimated by quantification of the IL-6 and collagen levels using ELISA and SIRCOL collagen assays, respectively. We found that S1P activated SUR2/Kir6.1 channel and that this effect was sensitive to specific inhibition of the S1P receptor of type 3 (S1P3R). In contrast, S1P1R receptor inhibition had no effect. Moreover, the S1P-induced current increased with cell culture duration whereas S1P3R expression level remained constant. The activation of SUR2/Kir6.1 channel by S1P via S1P3R stimulated cell proliferation and decreased IL-6 and collagen secretions. S1P also stimulated fibroblast migration via S1P3R but independently from SUR2/Kir6.1 channel activation. This study demonstrates that S1P, via S1P3R, affects cardiac ventricular fibroblasts function independently or through activation of SUR2/Kir6.1 channel. The latter effect occurs after fibroblasts differentiate into myofibroblasts, opening a new potential therapeutic strategy to modulate fibrosis after cardiac physiopathological injury.

  9. Disruption of the Candida albicans TPS1 Gene Encoding Trehalose-6-Phosphate Synthase Impairs Formation of Hyphae and Decreases Infectivity†

    Science.gov (United States)

    Zaragoza, Oscar; Blazquez, Miguel A.; Gancedo, Carlos

    1998-01-01

    The TPS1 gene from Candida albicans, which encodes trehalose-6-phosphate synthase, has been cloned by functional complementation of a tps1 mutant from Saccharomyces cerevisiae. In contrast with the wild-type strain, the double tps1/tps1 disruptant did not accumulate trehalose at stationary phase or after heat shock. Growth of the tps1/tps1 disruptant at 30°C was indistinguishable from that of the wild type. However, at 42°C it did not grow on glucose or fructose but grew normally on galactose or glycerol. At 37°C, the yeast-hypha transition in the mutant in glucose-calf serum medium did not occur. During growth at 42°C, the mutant did not form hyphae in galactose or in glycerol. Some of the growth defects observed may be traced to an unbalanced sugar metabolism that reduces the cellular content of ATP. Mice inoculated with 106 CFU of the tps1/tps1 mutant did not show visible symptoms of infection 16 days after inoculation, while those similarly inoculated with wild-type cells were dead 12 days after inoculation. PMID:9683476

  10. Influences of Dietary Added Sugar Consumption on Striatal Food-Cue Reactivity and Postprandial GLP-1 Response

    Directory of Open Access Journals (Sweden)

    Hilary M. Dorton

    2018-01-01

    Full Text Available Sugar consumption in the United States exceeds recommendations from the American Heart Association. Overconsumption of sugar is linked to risk for obesity and metabolic disease. Animal studies suggest that high-sugar diets alter functions in brain regions associated with reward processing, including the dorsal and ventral striatum. Human neuroimaging studies have shown that these regions are responsive to food cues, and that the gut-derived satiety hormones, glucagon-like peptide-1 (GLP-1, and peptide YY (PYY, suppress striatal food-cue responsivity. We aimed to determine the associations between dietary added sugar intake, striatal responsivity to food cues, and postprandial GLP-1 and PYY levels. Twenty-two lean volunteers underwent a functional magnetic resonance imaging (fMRI scan during which they viewed pictures of food and non-food items after a 12-h fast. Before scanning, participants consumed a glucose drink. A subset of 19 participants underwent an additional fMRI session in which they consumed water as a control condition. Blood was sampled for GLP-1, and PYY levels and hunger ratings were assessed before and ~75 min after drink consumption. In-person 24-h dietary recalls were collected from each participant on three to six separate occasions over a 2-month period. Average percent calories from added sugar were calculated using information from 24-h dietary recalls. A region-of-interest analysis was performed to compare the blood oxygen level-dependent (BOLD response to food vs. non-food cues in the bilateral dorsal striatum (caudate/putamen and ventral striatum (nucleus accumbens. The relationships between added sugar, striatal responses, and hormone changes after drink consumption were assessed using Spearman’s correlations. We observed a positive correlation between added sugar intake and BOLD response to food cues in the dorsal striatum and a similar trend in the nucleus accumbens after glucose, but not water, consumption

  11. Assessment of phosphate binding by sevelamer carbonate powder for oral suspension mixed in foods.

    Science.gov (United States)

    Hanus, Martin; Zhorov, Eugene; Brommage, Deborah; Plone, Melissa; Holmes-Farley, Stephen Randall

    2012-01-01

    This study investigated mixing sevelamer carbonate powder with foods and beverages other than water. Food samples, including applesauce, oatmeal, chicken, protein powder, scrambled eggs, ginger ale, and diet ginger ale, were subjected to an in vitro assay, and the difference in the amount of phosphate bound between samples pre-exposed to foods and samples where the drug was exposed to foods concurrently was determined Under these assay conditions, pre-exposure to sevelamer carbonate powder had no effect on the ability to bind phosphate. Clinical testing is needed to further evaluate this finding.

  12. Cardiovascular effects of sphingosine-1-phosphate and other sphingomyelin metabolites

    NARCIS (Netherlands)

    Alewijnse, Astrid E.; Peters, Stephan L. M.; Michel, Martin C.

    2004-01-01

    1 Upon various stimuli, cells metabolize sphingomyelin from the cellular plasma membrane to form sphingosylphosphorylcholine (SPC) or ceramide. The latter can be further metabolized to sphingosine and then sphingosine-1-phosphate (S1P). Apart from local formation, S1P and SPC are major constituents

  13. Filtration through nylon membranes negatively affects analysis of arsenic and phosphate by the molybdenum blue method

    DEFF Research Database (Denmark)

    Heimann, Axel Colin; Jakobsen, Rasmus

    2007-01-01

    Filtering synthetic arsenic- or phosphate-containing solutions (1.5-47.6 mu mol/L) with nylon syringe filters significantly reduced absorbances (by 6-74%) when analyzed with the colorimetric molybdenum blue method. Filtering the same solutions with cellulose acetate syringe filters yielded...... no significant differences as compared to unfiltered controls. The detrimental effect of nylon membranes was also observed when pure Milli-Q water was filtered and Subsequently spiked with arsenic(III) or phosphate suggesting that some compound(s) eluting from the filter membranes interfere with the color...... formation in the assay. Consequently, we caution against using nylon filters when filtering water samples for the determination of arsenic or phosphate with the molybdenum blue method....

  14. Phosphate concentration and arbuscular mycorrhizal colonisation influence the growth, yield and expression of twelve PHT1 family phosphate transporters in foxtail millet (Setaria italica.

    Directory of Open Access Journals (Sweden)

    S Antony Ceasar

    Full Text Available Phosphorus (P is an essential element which plays several key roles in all living organisms. Setaria italica (foxtail millet is a model species for panacoid grasses including several millet species widely grown in arid regions of Asia and Africa, and for the bioenergy crop switchgrass. The growth responses of S. italica to different levels of inorganic phosphate (Pi and to colonisation with the arbuscular mycorrhizal fungus Funneliformis mosseae (syn. Glomus mosseae were studied. Phosphate is taken up from the environment by the PHT1 family of plant phosphate transporters, which have been well characterized in several plant species. Bioinformatic analysis identified 12 members of the PHT1 gene family (SiPHT1;1-1;12 in S. italica, and RT and qPCR analysis showed that most of these transporters displayed specific expression patterns with respect to tissue, phosphate status and arbuscular mycorrhizal colonisation. SiPHT1;2 was found to be expressed in all tissues and in all growth conditions tested. In contrast, expression of SiPHT1;4 was induced in roots after 15 days growth in hydroponic medium of low Pi concentration. Expression of SiPHT1;8 and SiPHT1;9 in roots was selectively induced by colonisation with F. mosseae. SiPHT1;3 and SiPHT1;4 were found to be predominantly expressed in leaf and root tissues respectively. Several other transporters were expressed in shoots and leaves during growth in low Pi concentrations. This study will form the basis for the further characterization of these transporters, with the long term goal of improving the phosphate use efficiency of foxtail millet.

  15. Phosphate concentration and arbuscular mycorrhizal colonisation influence the growth, yield and expression of twelve PHT1 family phosphate transporters in foxtail millet (Setaria italica).

    Science.gov (United States)

    Ceasar, S Antony; Hodge, Angela; Baker, Alison; Baldwin, Stephen A

    2014-01-01

    Phosphorus (P) is an essential element which plays several key roles in all living organisms. Setaria italica (foxtail millet) is a model species for panacoid grasses including several millet species widely grown in arid regions of Asia and Africa, and for the bioenergy crop switchgrass. The growth responses of S. italica to different levels of inorganic phosphate (Pi) and to colonisation with the arbuscular mycorrhizal fungus Funneliformis mosseae (syn. Glomus mosseae) were studied. Phosphate is taken up from the environment by the PHT1 family of plant phosphate transporters, which have been well characterized in several plant species. Bioinformatic analysis identified 12 members of the PHT1 gene family (SiPHT1;1-1;12) in S. italica, and RT and qPCR analysis showed that most of these transporters displayed specific expression patterns with respect to tissue, phosphate status and arbuscular mycorrhizal colonisation. SiPHT1;2 was found to be expressed in all tissues and in all growth conditions tested. In contrast, expression of SiPHT1;4 was induced in roots after 15 days growth in hydroponic medium of low Pi concentration. Expression of SiPHT1;8 and SiPHT1;9 in roots was selectively induced by colonisation with F. mosseae. SiPHT1;3 and SiPHT1;4 were found to be predominantly expressed in leaf and root tissues respectively. Several other transporters were expressed in shoots and leaves during growth in low Pi concentrations. This study will form the basis for the further characterization of these transporters, with the long term goal of improving the phosphate use efficiency of foxtail millet.

  16. Sphingosine 1-phosphate and cancer.

    Science.gov (United States)

    Pyne, Nigel J; El Buri, Ashref; Adams, David R; Pyne, Susan

    2017-09-15

    The bioactive lipid, sphingosine 1-phosphate (S1P) is produced by phosphorylation of sphingosine and this is catalysed by two sphingosine kinase isoforms (SK1 and SK2). Here we discuss structural functional aspects of SK1 (which is a dimeric quaternary enzyme) that relate to coordinated coupling of membrane association with phosphorylation of Ser225 in the 'so-called' R-loop, catalytic activity and protein-protein interactions (e.g. TRAF2, PP2A and G q ). S1P formed by SK1 at the plasma-membrane is released from cells via S1P transporters to act on S1P receptors to promote tumorigenesis. We discuss here an additional novel mechanism that can operate between cancer cells and fibroblasts and which involves the release of the S1P receptor, S1P 2 in exosomes from breast cancer cells that regulates ERK-1/2 signalling in fibroblasts. This novel mechanism of signalling might provide an explanation for the role of S1P 2 in promoting metastasis of cancer cells and which is dependent on the micro-environmental niche. Copyright © 2017. Published by Elsevier Ltd.

  17. Alpha 1 B- but not alpha 1 A-adrenoceptors mediate inositol phosphate generation

    NARCIS (Netherlands)

    Michel, M. C.; Hanft, G.; Gross, G.

    1990-01-01

    We used novel highly subtype-selective antagonists to study whether alpha 1A- and/or alpha 1B-adrenoceptors mediate the stimulation of inositol phosphate generation by noradrenaline in rat cerebral cortex. Phentolamine (10 microM) and prazosin (100 nM) completely abolished the stimulated inositol

  18. Evaluation of P availability from Fe and A1 labelled (32P) phosphates

    International Nuclear Information System (INIS)

    Bittencourt, V.S.

    1975-07-01

    Synthetically Fe and A1 labelled phosphates ( 32 P) show a certain amount of available P to the plants when applied to Sao Paulo State soils. This availability decreases from considered amorphous A1-phosphate (A1-P sub(am)) to A1 phosphate with a certain cristalinity grade (A1-P sub(cr)) and than from this to Fe-P sub(am) followed by Fe-P sub(cr), and it is influenced by both the soil characteristics and mainly by the iron constituents of the samples. In this way, one can not expect that the 0,05 N 2 H SO 4 and the CHANG and JACKSON (1957a) solutions can define properly the available P of these soils. The addition of lime to the soils do not drive to a better P absorption by the plants and its effects are dubious

  19. Sphingosine 1-phosphate as a novel immune regulator of dendritic ...

    Indian Academy of Sciences (India)

    Although originally described as an intracellular second messenger, sphingosine 1-phosphate (S1P) has recently been shown to be involved in several physiological and pathological functions as an extracellular mediator. S1P receptors are widely expressed and thought to regulate important functions in cell signalling.

  20. Synthesis, structure and luminescence properties of phosphates A1 ...

    Indian Academy of Sciences (India)

    rule and the localization of this cation in the structure, which is in agreement with the structural characterization. It appears that europium ... 1. Introduction. Atom isomorphism in complex crystalline compounds with ..... isotropic approx- imation of basis atom positions in the structures of phosphates Na0.7Eu0.1Zr2(PO4)3 and.

  1. Dielectric properties of residual water in amorphous lyophilized mixtures of sugar and drug

    Energy Technology Data Exchange (ETDEWEB)

    Moznine, R El [School of Pharmacy, De Montfort University, Leiceste (United Kingdom); Smith, G [School of Pharmacy, De Montfort University, Leicester (United Kingdom); Polygalov, E [School of Pharmacy, De Montfort University, Leicester (United Kingdom); Suherman, P M [School of Pharmacy, De Montfort University, Leicester (United Kingdom); Broadhead, J [AstraZeneca Charnwood R and D, Bakewell Rd, Loughborough (United Kingdom)

    2003-02-21

    Dielectric relaxation spectroscopy was used to investigate the properties of residual water in lyophilized formulations of a proprietary tri-phosphate drug containing a sugar (trehalose, lactose or sucrose) or dextran. The dielectric properties of each formulation were determined in the frequency range (0.1 Hz-0.1 MHz) and temperature range (30 deg. C-T{sub g}). The temperature dependence of the relaxation times for all samples showed Arrhenuis behaviour, from which the activation energy was derived. Proton hopping through the hydrogen-bonded network (clusters) of water molecules was suggested as the principle mode of charge transport. Significant differences in dielectric relaxation kinetics and activation energy were observed for the different formulations, which were found to correlate with the amount of monophosphate degradation product.

  2. Sphingosine-1-Phosphate Signaling in Inflammatory Bowel Disease

    DEFF Research Database (Denmark)

    Nielsen, Ole Haagen; Li, Yuan; Lindbom, Bengt Johansson

    2017-01-01

    primary non-responders or lose responsiveness during maintenance treatment. A new class of small molecules, sphingosine-1-phosphate (S1P) receptor modulators, has recently shown efficacy in IBD. Here we provide an overview of the mechanism of action of this novel treatment principle in the context...... of intestinal inflammation. The remarkable impact of therapeutic modulation of the S1P/S1P receptor axis reflects the complexity of the pathogenesis of IBD and the fact that S1P receptor modulation may be a logical therapeutic approach for the future management of IBD....

  3. Engineering and exploitation of a fluorescent HIV-1 gp120 for live cell CD4 binding assays

    Energy Technology Data Exchange (ETDEWEB)

    Costantini, Lindsey M. [Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461 (United States); Irvin, Susan C. [Department of Pediatrics, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461 (United States); Department of Microbiology and Immunology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461 (United States); Kennedy, Steven C. [Department of Pediatrics, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461 (United States); Guo, Feng [Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461 (United States); Goldstein, Harris; Herold, Betsy C. [Department of Pediatrics, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461 (United States); Department of Microbiology and Immunology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461 (United States); Snapp, Erik L., E-mail: erik-lee.snapp@einstein.yu.edu [Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461 (United States)

    2015-02-15

    The HIV-1 envelope glycoprotein, gp120, binds the host cell receptor, CD4, in the initial step of HIV viral entry and infection. This process is an appealing target for the development of inhibitory drugs and neutralizing antibodies. To study gp120 binding and intracellular trafficking, we engineered a fluorescent fusion of the humanized gp120 JRFL HIV-1 variant and GFP. Gp120-sfGFP is glycosylated with human sugars, robustly expressed, and secreted from cultured human cells. Protein dynamics, quality control, and trafficking can be visualized in live cells. The fusion protein can be readily modified with different gp120 variants or fluorescent proteins. Finally, secreted gp120-sfGFP enables a sensitive and easy binding assay that can quantitatively screen potential inhibitors of gp120-CD4 binding on live cells via fluorescence imaging or laser scanning cytometry. This adaptable research tool should aid in studies of gp120 cell biology and the development of novel anti-HIV drugs. - Highlights: • Development of fluorescent protein labeled HIV-1 envelope gp120. • Imaging of gp120 dynamics and trafficking in live cells. • Quantitative visual assay of antibody-mediated inhibition of gp120 binding to CD4 on live cells.

  4. Engineering and exploitation of a fluorescent HIV-1 gp120 for live cell CD4 binding assays

    International Nuclear Information System (INIS)

    Costantini, Lindsey M.; Irvin, Susan C.; Kennedy, Steven C.; Guo, Feng; Goldstein, Harris; Herold, Betsy C.; Snapp, Erik L.

    2015-01-01

    The HIV-1 envelope glycoprotein, gp120, binds the host cell receptor, CD4, in the initial step of HIV viral entry and infection. This process is an appealing target for the development of inhibitory drugs and neutralizing antibodies. To study gp120 binding and intracellular trafficking, we engineered a fluorescent fusion of the humanized gp120 JRFL HIV-1 variant and GFP. Gp120-sfGFP is glycosylated with human sugars, robustly expressed, and secreted from cultured human cells. Protein dynamics, quality control, and trafficking can be visualized in live cells. The fusion protein can be readily modified with different gp120 variants or fluorescent proteins. Finally, secreted gp120-sfGFP enables a sensitive and easy binding assay that can quantitatively screen potential inhibitors of gp120-CD4 binding on live cells via fluorescence imaging or laser scanning cytometry. This adaptable research tool should aid in studies of gp120 cell biology and the development of novel anti-HIV drugs. - Highlights: • Development of fluorescent protein labeled HIV-1 envelope gp120. • Imaging of gp120 dynamics and trafficking in live cells. • Quantitative visual assay of antibody-mediated inhibition of gp120 binding to CD4 on live cells

  5. Sphingosine 1-phosphate induces neutrophil chemoattractant IL-8: repression by steroids.

    Directory of Open Access Journals (Sweden)

    Md Mostafizur Rahman

    Full Text Available The bioactive sphingolipid sphingosine 1-phosphate (S1P is found in increased amounts in the airways of asthmatics. S1P can regulate airway smooth muscle functions associated with asthmatic inflammation and remodeling, including cytokine secretion. To date however, whether S1P induces secretion of an important chemokine responsible for neutrophilia in airway inflammation--IL-8--was unexplored. The aim of this study was to investigate whether S1P induces IL-8 gene expression and secretion to enhance neutrophil chemotaxis in vitro, as well as examine the molecular mechanisms responsible for repression by the corticosteroid dexamethasone. We show that S1P upregulates IL-8 secretion from ASM cells and enhance neutrophil chemotaxis in vitro. The corticosteroid dexamethasone significantly represses IL-8 mRNA expression and protein secretion in a concentration- and time-dependent manner. Additionally, we reveal that S1P-induced IL-8 secretion is p38 MAPK and ERK-dependent and that these key phosphoproteins act on the downstream effector mitogen- and stress-activated kinase 1 (MSK1 to control secretion of the neutrophil chemoattractant cytokine IL-8. The functional relevance of this in vitro data was demonstrated by neutrophil chemotaxis assays where S1P-induced effects can be significantly attenuated by pretreatment with dexamethasone, pharmacological inhibition of p38 MAPK- or ERK-mediated pathways, or by knocking down MSK-1 with siRNA. Taken together, our study reveals the molecular pathways responsible for IL-8 secretion from ASM cells in response to S1P and indicates ways in which the impact on IL-8-driven neutrophilia may be lessened.

  6. Preparation of Reducing Sugar Hydrolyzed from High-Lignin Coconut Coir Dust Pretreated by the Recycled Ionic Liquid [mmim][dmp] and Combination with Alkaline

    Directory of Open Access Journals (Sweden)

    Hanny Frans Sangian

    2015-03-01

    Full Text Available This study aims to produce reducing sugar hydrolyzed from substrate, coconut coir dust pretreated by recycled ionic liquid and its combination with alkaline. The 1H NMR and FTIR were performed to ver-ify the synthesized ionic liquid methylmethylimidazolium dimethyl phosphate ([mmim][dmp]. The structure of pretreated substrates was analyzed by XRD measurement. The used ionic liquid was recy-cled twice to re-employ for substrate pretreatment. The treated- and untreated-coconut coir dust were hydrolyzed into sugars using pure cellulase. The reaction, which called an enzymatic hydrolysis, was conducted at 60 °C, pH 3, for 48 h. The yields of sugar hydrolyzed from fresh IL-pretreated, 1R*IL-pretreated and 2R*IL-pretreated substrates were of 0.19, 0.15 and 0.15 g sugar / g cellu-lose+hemicellulose, respectively. Pretreatment with NaOH or the combination of NaOH+IL resulted in yields of reducing sugars of 0.25, 0.28 g/g, respectively. When alkaline combined with the recycled ionic liquids, NaOH+1R*IL, NaOH+2R*IL in the pretreatment, the yields of sugar were relatively similar to those obtained using alkaline followed by fresh ionic liquid. If the mixture enzymes, cellu-lase+xylanase, used to liberate sugars from fresh IL-pretreated, or recycled IL-pretreated substrates, the amount of sugar (concentration or yield increased slightly compared to that employing a single cel-lulase. These findings showed that recycled IL pretreatment of the high-lignin lignocellulose, coconut coir dust, is a new prospect for the economical manufacture of fermentable sugars and biofuel in the coming years. © 2015 BCREC UNDIP. All rights reservedReceived: 1st July 2014; Revised: 5th September 2014; Accepted: 5th September 2014 How to Cite: Sangian, H.F., Kristian, J., Rahma, S., Dewi, H., Puspasari, D., Agnesty, S., Gunawan, S., Widjaja, A. (2015. Preparation of Reducing Sugar Hydrolyzed from High-Lignin Coconut Coir Dust Pretreated by the Recycled Ionic Liquid [mmim

  7. Sugar holograms with erioglaucine and tartrazine

    Science.gov (United States)

    Mejias-Brizuela, N. Y.; Olivares-Pérez, A.; Páez-Trujillo, G.; Fuentes-Tapia, I.

    2007-09-01

    An artificial green colorant, composed by erioglaucine (Blue 1) and tartrazine (Yellow 5), was employed in a sugar matrix to improve the material sensibility and to make a comparative analysis of the diffraction efficiency parameter, for holograms replications, the holographic pattern was obtained by a computer and recorded in sugar films and in modified sugar (sugar-colorant). Conventional lithography and UV radiation were used. The results show that the behavior diffraction efficiency of the sugar-colorant films is slightly larger than in the sugar matrix under the same recording conditions.

  8. A four-gene operon in Bacillus cereus produces two rare spore-decorating sugars.

    Science.gov (United States)

    Li, Zi; Mukherjee, Thiya; Bowler, Kyle; Namdari, Sholeh; Snow, Zachary; Prestridge, Sarah; Carlton, Alexandra; Bar-Peled, Maor

    2017-05-05

    Bacterial glycan structures on cell surfaces are critical for cell-cell recognition and adhesion and in host-pathogen interactions. Accordingly, unraveling the sugar composition of bacterial cell surfaces can shed light on bacterial growth and pathogenesis. Here, we found that two rare sugars with a 3- C -methyl-6-deoxyhexose structure were linked to spore glycans in Bacillus cereus ATCC 14579 and ATCC 10876. Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM: C -methyltransferase, and NADPH-dependent CDP-3- C -methyl-6-deoxyhexose 4-reductase. The last enzyme predominantly yielded CDP-3- C -methyl-6-deoxygulose (CDP-cereose) and likely generated a 4-epimer CDP-3- C -methyl-6-deoxyallose (CDP-cillose). Some members of the B. cereus sensu lato group produce CDP-3- C -methyl-6-deoxy sugars for the formation of cereose-containing glycans on spores, whereas others such as Bacillus anthracis do not. Gene knockouts of the Bacillus C -methyltransferase and the 4-reductase confirmed their involvement in the formation of cereose-containing glycan on B. cereus spores. We also found that cereose represented 0.2-1% spore dry weight. Moreover, mutants lacking cereose germinated faster than the wild type, yet the mutants exhibited no changes in sporulation or spore resistance to heat. The findings reported here may provide new insights into the roles of the uncommon 3- C -methyl-6-deoxy sugars in cell-surface recognition and host-pathogen interactions of the genus Bacillus . © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  9. Substrate scope of a dehydrogenase from Sphingomonas species A1 and its potential application in the synthesis of rare sugars and sugar derivatives.

    Science.gov (United States)

    Beer, Barbara; Pick, André; Döring, Manuel; Lommes, Petra; Sieber, Volker

    2018-04-26

    Rare sugars and sugar derivatives that can be obtained from abundant sugars are of great interest to biochemical and pharmaceutical research. Here, we describe the substrate scope of a short-chain dehydrogenase/reductase from Sphingomonas species A1 (SpsADH) in the oxidation of aldonates and polyols. The resulting products are rare uronic acids and rare sugars respectively. We provide insight into the substrate recognition of SpsADH using kinetic analyses, which show that the configuration of the hydroxyl groups adjacent to the oxidized carbon is crucial for substrate recognition. Furthermore, the specificity is demonstrated by the oxidation of d-sorbitol leading to l-gulose as sole product instead of a mixture of d-glucose and l-gulose. Finally, we applied the enzyme to the synthesis of l-gulose from d-sorbitol in an in vitro system using a NADH oxidase for cofactor recycling. This study shows the usefulness of exploring the substrate scope of enzymes to find new enzymatic reaction pathways from renewable resources to value-added compounds. © 2018 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  10. The Effect of Sugar Price Policy on U.S. Imports of Processed Sugar-containing foods

    OpenAIRE

    Jabara, Cathy L.

    1988-01-01

    This paper examines the effects of sugar protection, as well as the effects of these other factors, on U.S. demand for imports of sugar-containing products. The paper also addresses two main issues: (1) whether substitution of alternative sweeteners has allowed U.S. food manufactures to reduce the competitive advantage provided to foreign manufactures by U.S. sugar policies, and (2) which countries -- developed or developing -- have been able to increase their exports of sugar-containing prod...

  11. Exposure to tri-o-cresyl phosphate detected in jet airplane passengers

    International Nuclear Information System (INIS)

    Liyasova, Mariya; Li, Bin; Schopfer, Lawrence M.; Nachon, Florian; Masson, Patrick; Furlong, Clement E.; Lockridge, Oksana

    2011-01-01

    The aircraft cabin and flight deck ventilation are supplied from partially compressed unfiltered bleed air directly from the engine. Worn or defective engine seals can result in the release of engine oil into the cabin air supply. Aircrew and passengers have complained of illness following such “fume events”. Adverse health effects are hypothesized to result from exposure to tricresyl phosphate mixed esters, a chemical added to jet engine oil and hydraulic fluid for its anti-wear properties. Our goal was to develop a laboratory test for exposure to tricresyl phosphate. The assay was based on the fact that the active-site serine of butyrylcholinesterase reacts with the active metabolite of tri-o-cresyl phosphate, cresyl saligenin phosphate, to make a stable phosphorylated adduct with an added mass of 80 Da. No other organophosphorus agent makes this adduct in vivo on butyrylcholinesterase. Blood samples from jet airplane passengers were obtained 24–48 h after completing a flight. Butyrylcholinesterase was partially purified from 25 ml serum or plasma, digested with pepsin, enriched for phosphorylated peptides by binding to titanium oxide, and analyzed by mass spectrometry. Of 12 jet airplane passengers tested, 6 were positive for exposure to tri-o-cresyl phosphate that is, they had detectable amounts of the phosphorylated peptide FGEpSAGAAS. The level of exposure was very low. No more than 0.05 to 3% of plasma butyrylcholinesterase was modified. None of the subjects had toxic symptoms. Four of the positive subjects were retested 3 to 7 months following their last airplane trip and were found to be negative for phosphorylated butyrylcholinesterase. In conclusion, this is the first report of an assay that detects exposure to tri-o-cresyl phosphate in jet airplane travelers. -- Highlights: ► Travel on jet airplanes is associated with an illness, aerotoxic syndrome. ► A possible cause is exposure to tricresyl phosphate in engine lubricating oil. ► A blood

  12. Exposure to tri-o-cresyl phosphate detected in jet airplane passengers

    Energy Technology Data Exchange (ETDEWEB)

    Liyasova, Mariya, E-mail: mliyasov@unmc.edu [Eppley Institute, University of Nebraska Medical Center, Omaha, NE 68198-5950 (United States); Department of Environmental, Agricultural, and Occupational Health, University of Nebraska Medical Center, Omaha, NE (United States); Li, Bin, E-mail: binli@unmc.edu [Eppley Institute, University of Nebraska Medical Center, Omaha, NE 68198-5950 (United States); Schopfer, Lawrence M., E-mail: lmschopf@unmc.edu [Eppley Institute, University of Nebraska Medical Center, Omaha, NE 68198-5950 (United States); Nachon, Florian, E-mail: fnachon@nachon.net [Departement de Toxicologie, Institut de Recherche Biomedicale des Armees, 24 avenue des Marquis du Gresivaudan, 38702 La Tronche (France); Masson, Patrick, E-mail: pmasson@unmc.edu [Departement de Toxicologie, Institut de Recherche Biomedicale des Armees, 24 avenue des Marquis du Gresivaudan, 38702 La Tronche (France); Eppley Institute, University of Nebraska Medical Center, Omaha, NE 68198-5950 (United States); Furlong, Clement E., E-mail: clem@uw.edu [Department of Medicine (Div. Medical Genetics) and Genome Sciences, University of Washington, Seattle, WA 98195 (United States); Lockridge, Oksana, E-mail: olockrid@unmc.edu [Eppley Institute, University of Nebraska Medical Center, Omaha, NE 68198-5950 (United States); Department of Environmental, Agricultural, and Occupational Health, University of Nebraska Medical Center, Omaha, NE (United States)

    2011-11-15

    The aircraft cabin and flight deck ventilation are supplied from partially compressed unfiltered bleed air directly from the engine. Worn or defective engine seals can result in the release of engine oil into the cabin air supply. Aircrew and passengers have complained of illness following such 'fume events'. Adverse health effects are hypothesized to result from exposure to tricresyl phosphate mixed esters, a chemical added to jet engine oil and hydraulic fluid for its anti-wear properties. Our goal was to develop a laboratory test for exposure to tricresyl phosphate. The assay was based on the fact that the active-site serine of butyrylcholinesterase reacts with the active metabolite of tri-o-cresyl phosphate, cresyl saligenin phosphate, to make a stable phosphorylated adduct with an added mass of 80 Da. No other organophosphorus agent makes this adduct in vivo on butyrylcholinesterase. Blood samples from jet airplane passengers were obtained 24-48 h after completing a flight. Butyrylcholinesterase was partially purified from 25 ml serum or plasma, digested with pepsin, enriched for phosphorylated peptides by binding to titanium oxide, and analyzed by mass spectrometry. Of 12 jet airplane passengers tested, 6 were positive for exposure to tri-o-cresyl phosphate that is, they had detectable amounts of the phosphorylated peptide FGEpSAGAAS. The level of exposure was very low. No more than 0.05 to 3% of plasma butyrylcholinesterase was modified. None of the subjects had toxic symptoms. Four of the positive subjects were retested 3 to 7 months following their last airplane trip and were found to be negative for phosphorylated butyrylcholinesterase. In conclusion, this is the first report of an assay that detects exposure to tri-o-cresyl phosphate in jet airplane travelers. -- Highlights: Black-Right-Pointing-Pointer Travel on jet airplanes is associated with an illness, aerotoxic syndrome. Black-Right-Pointing-Pointer A possible cause is exposure to

  13. The Properties of Sintered Calcium Phosphate with [Ca]/[P] = 1.50

    Directory of Open Access Journals (Sweden)

    Moo-Chin Wang

    2012-10-01

    Full Text Available In order to obtain the properties of the sintered as-dried calcium phosphate with [Ca]/[P] = 1.50, the characteristics of sintered pellets have been investigated using X-ray diffraction (XRD, inductively coupled plasma-mass spectrometry (ICP-MS, Fourier-transform infrared (FT-IR spectra, Vickers hardness indentation and scanning electron microscopy (SEM. When the pellet samples were sintered between 700 °C and 1200 °C for 4 h, the hydroxyapatite (Ca10(PO46(OH2, HA still maintained the major phase, accompanied with the rhenanite (NaCaPO4 as the secondary phase and β-tricalcium phosphate (β-Ca3(PO42, β-TCP as the minor phases. In addition, the HA partially transformed to α-tricalcium phosphate (α-Ca3(PO42, α-TCP and tetracalcium phosphate (Ca4(PO42O, TTCP, when the pellet samples were sintered at 1300 °C and 1400 °C, respectively, for 4 h. The maximum density and Vickers Hardness (HV of sintered pellet samples were 2.85 g/cm3 (90.18% theoretical density (T.D. and 407, which appeared at 1200 °C and 900 °C, respectively.

  14. Sphingosine Kinase 1 and Sphingosine-1-Phosphate Signaling in Colorectal Cancer

    Directory of Open Access Journals (Sweden)

    Yonghua Bao

    2017-10-01

    Full Text Available Sphingosine kinase 1 (Sphk1 is a highly conserved lipid kinase that phosphorylates sphingosine to form sphingosine-1-phosphate (S1P. Growing studies have demonstrated that Sphk1 is overexpressed in various types of solid cancers and can be induced by growth factors, cytokines, and carcinogens, leading to the increase of S1P production. Subsequently, the increased Sphk1/S1P facilitates cancer cell proliferation, mobility, angiogenesis, invasion, and metastasis. Therefore, Sphk1/S1P signaling plays oncogenic roles. This review summarizes the features of Sphk1/S1P signaling and their functions in colorectal cancer cell growth, tumorigenesis, and metastasis, as well as the possible underlying mechanisms.

  15. A rapid method for simultaneous quantification of 13 sugars and sugar alcohols in food products by UPLC-ELSD.

    Science.gov (United States)

    Koh, Dong-Wan; Park, Jae-Woong; Lim, Jung-Hoon; Yea, Myeong-Jai; Bang, Dae-Young

    2018-02-01

    A novel, rapid, simultaneous analysis method for five sugars (fructose, glucose, sucrose, maltose, and lactose) and eight sugar alcohols (erythritol, xylitol, sorbitol, mannitol, inositol, maltitol, lactitol, and isomalt) was developed using UPLC-ELSD, without derivatization. The analysis conditions, including the gradient conditions, modifier concentration and column length, were optimized. Thirteen sugars and sugar alcohols were separated well and the resolution of their peaks was above 1.0. Their optimum analysis condition can be analyzed within 15min. Standard curves for sugars and sugar alcohols with concentrations of 5.0-0.1% and 2.0-0.05% are presented herein, and their correlation coefficients are found to be above 0.999 and the limit of detection (LOD) was around 0.006-0.018%. This novel analysis system can be used for foodstuffs such as candy, chewing gum, jelly, chocolate, processed chocolate products, and snacks containing 0.21-46.41% of sugars and sugar alcohols. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. 31P-Nuclear Magnetic Resonance Determination of Phosphate Compartmentation in Leaves of Reproductive Soybeans (Glycine max L.) as Affected by Phosphate Nutrition 1

    Science.gov (United States)

    Lauer, Michael J.; Blevins, Dale G.; Sierzputowska-Gracz, Hanna

    1989-01-01

    Most leaf phosphorus is remobilized to the seed during reproductive development in soybean. We determined, using 31P-NMR, the effect phosphorus remobilization has on vacuolar inorganic phosphate pool size in soybean (Glycine max [L.] Merr.) leaves with respect to phosphorus nutrition and plant development. Phosphate compartmentation between cytoplasmic and vacuolar pools was observed and followed in intact tissue grown hydroponically, at the R2, R4, and R6 growth stages. As phosphorus in the nutrient solution decreased from 0.45 to 0.05 millimolar, the vacuolar phosphate peak became less prominent relative to cytoplasmic phosphate and hexose monophosphate peaks. At a nutrient phosphate concentration of 0.05 millimolar, the vacuolar phosphate peak was not detectable. At higher levels of nutrient phosphate, as plants progressed from the R2 to the R6 growth stage, the vacuolar phosphate peak was the first to disappear, suggesting that storage phosphate was remobilized to a greater extent than metabolic phosphate. Under suboptimal phosphate nutrition (≤ 0.20 millimolar), the hexose monophosphate and cytoplasmic phosphate peaks declined earlier in reproductive development than when phosphate was present in optimal amounts. Under low phosphate concentrations (0.05 millimolar) cytoplasmic phosphate was greatly reduced. Carbon metabolism was coincidently disrupted under low phosphate nutrition as shown by the appearance of large, prominent starch grains in the leaves. Cytoplasmic phosphate, and leaf carbon metabolism dependent on it, are buffered by vacuolar phosphate until late stages of reproductive growth. Images Figure 4 PMID:16666705

  17. Phosphorus in chronosequence of burnt sugar cane in Brazilian cerrado: humic acid analysis by {sup 31}P NMR; Fosforo em cronossequencia de cana-de-acucar queimada no cerrado goiano: analise de acidos humicos por RMN de {sup 31}P

    Energy Technology Data Exchange (ETDEWEB)

    Rossi, Celeste Q.; Pereira, Marcos G.; Garcia, Andreas C., E-mail: mgervasiopereira@gmail.com [Universidade Federal Rural do Rio de Janeiro (UFRRJ), Seropedica, RJ (Brazil). Dept. de Solos; Perin, Adriano; Gazolla, Paulo R. [Instituto Federal de Educacao, Ciencia e Tecnologia Goiano, Rio Verde, GO (Brazil); Gonzalez, Antonio P. [Universidade de Coruna, ES (Spain). Faculdad de Ciencias

    2013-10-01

    The aim of this study was to identify, with the use of {sup 31}P NMR spectroscopy, organic P species in humic acids (HA) in samples from Oxisol cultivated in chronosequence with sugar cane, pasture and Cerrado. The main forms of P-type found were orthophosphate, monoester-P (phosphate sugars) and P-diester (orthophosphate). The {sup 31}P NMR technique proved capable of identifying changes in the areas studied as a function of sugar cane burning time. In areas with 1 and 5 years of burnt cane, a decrease in recalcitrant organic P in humic acids indicated the need for use of P-humic substances for plant nutrition (author)

  18. Active site of Zn2+-dependent sn-glycerol-1-phosphate dehydrogenase from Aeropyrum pernix K1

    Directory of Open Access Journals (Sweden)

    Jin-Suk Han

    2005-01-01

    Full Text Available The enzyme sn-glycerol-1-phosphate dehydrogenase (Gro1PDH, EC 1.1.1.261 is key to the formation of the enantiomeric configuration of the glycerophosphate backbone (sn-glycerol-1-phosphate of archaeal ether lipids. This enzyme catalyzes the reversible conversion between dihydroxyacetone phosphate and glycerol-1-phosphate. To date, no information about the active site and catalytic mechanism of this enzyme has been reported. Using the sequence and structural information for glycerol dehydrogenase, we constructed six mutants (D144N, D144A, D191N, H271A, H287A and D191N/H271A of Gro1PDH from Aeropyrum pernix K1 and examined their characteristics to clarify the active site of this enzyme. The enzyme was found to be a zinc-dependent metalloenzyme, containing one zinc ion for every monomer protein that was essential for activity. Site-directed mutagenesis of D144 increased the activity of the enzyme. Mutants D144N and D144A exhibited low affinity for the substrates and higher activity than the wild type, but their affinity for the zinc ion was the same as that of the wild type. Mutants D191N, H271A and H287A had a low affinity for the zinc ion and a low activity compared with the wild type. The double mutation, D191N/ H271A, had no enzyme activity and bound no zinc. From these results, it was clarified that residues D191, H271 and H287 participate in the catalytic activity of the enzyme by binding the zinc ion, and that D144 has an effect on substrate binding. The structure of the active site of Gro1PDH from A. pernix K1 seems to be similar to that of glycerol dehydrogenase, despite the differences in substrate specificity and biological role.

  19. Phosphate transporter mediated lipid accumulation in Saccharomyces cerevisiae under phosphate starvation conditions.

    Science.gov (United States)

    James, Antoni W; Nachiappan, Vasanthi

    2014-01-01

    In the current study, when phosphate transporters pho88 and pho86 were knocked out they resulted in significant accumulation (84% and 43%) of triacylglycerol (TAG) during phosphate starvation. However in the presence of phosphate, TAG accumulation was only around 45% in both pho88 and pho86 mutant cells. These observations were confirmed by radio-labeling, fluorescent microscope and RT-PCR studies. The TAG synthesizing genes encoding for acyltransferases namely LRO1 and DGA1 were up regulated. This is the first report for accumulation of TAG in pho88Δ and pho86Δ cells under phosphate starvation conditions. Copyright © 2013. Published by Elsevier Ltd.

  20. Human-Phosphate-Binding-Protein inhibits HIV-1 gene transcription and replication

    Directory of Open Access Journals (Sweden)

    Candolfi Ermanno

    2011-07-01

    Full Text Available Abstract The Human Phosphate-Binding protein (HPBP is a serendipitously discovered lipoprotein that binds phosphate with high affinity. HPBP belongs to the DING protein family, involved in various biological processes like cell cycle regulation. We report that HPBP inhibits HIV-1 gene transcription and replication in T cell line, primary peripherical blood lymphocytes and primary macrophages. We show that HPBP is efficient in naïve and HIV-1 AZT-resistant strains. Our results revealed HPBP as a new and potent anti HIV molecule that inhibits transcription of the virus, which has not yet been targeted by HAART and therefore opens new strategies in the treatment of HIV infection.

  1. Food acid content and erosive potential of sugar-free confections.

    Science.gov (United States)

    Shen, P; Walker, G D; Yuan, Y; Reynolds, C; Stacey, M A; Reynolds, E C

    2017-06-01

    Dental erosion is an increasingly prevalent problem associated with frequent consumption of acidic foods and beverages. The aim of this study was to measure the food acid content and the erosive potential of a variety of sugar-free confections. Thirty sugar-free confections were selected and extracts analysed to determine pH, titratable acidity, chemical composition and apparent degree of saturation with respect to apatite. The effect of the sugar-free confections in artificial saliva on human enamel was determined in an in vitro dental erosion assay using change in surface microhardness. The change in surface microhardness was used to categorize the confections as high, moderate or low erosive potential. Seventeen of the 30 sugar-free confections were found to contain high concentrations of food acids, exhibit low pH and high titratable acidity and have high erosive potential. Significant correlations were found between the dental erosive potential (change in enamel surface microhardness) and pH and titratable acidity of the confections. Ten of these high erosive potential confections displayed dental messages on the packaging suggesting they were safe for teeth. Many sugar-free confections, even some with 'Toothfriendly' messages on the product label, contain high contents of food acids and have erosive potential. © 2017 Australian Dental Association.

  2. Antagonism of the morphine-induced locomotor activation of mice by fructose: comparison with other opiates and sugars, and sugar effects on brain morphine.

    Science.gov (United States)

    Brase, D A; Ward, C R; Bey, P S; Dewey, W L

    1991-01-01

    The mouse locomotor activation test of opiate action in a 2+2 dose parallel line assay was used in a repeated testing paradigm to determine the test, opiate and hexose specificities of a previously reported antagonism of morphine-induced antinocociception by hyperglycemia. In opiate specificity studies, fructose (5 g/kg, i.p.) significantly reduced the potency ratio for morphine and methadone, but not for levorphanol, meperidine or phenazocine when intragroup comparisons were made. In intergroup comparisons, fructose significantly reduced the potencies of levorphanol and phenazocine, but not methadone or meperidine. In hexose/polyol specificity studies, tagatose and fructose significantly reduced the potency ratio for morphine, whereas glucose, galactose, mannose and the polyols, sorbitol and xylitol, caused no significant decrease in potency. Fructose, tagatose, glucose and mannose (5 g/kg, i.p.) were tested for effects on brain morphine levels 30 min after morphine (60 min after sugar), and all four sugars significantly increased brain morphine relative to saline-pretreated controls. It is concluded that the antagonism of morphine by acute sugar administration shows specificity for certain sugars and occurs despite sugar-induced increases in the distribution of morphine to the brain. Furthermore, the effects of fructose show an opiate specificity similar to that of glucose on antinociception observed previously in our laboratory, except that methadone was also significantly inhibited in the present study, when a repeated-testing experimental design was used.

  3. Sugar Sugar – don’t be misled / laat je niet misleiden

    NARCIS (Netherlands)

    Toebes, Brigit

    2017-01-01

    NRC Handelsblad’s Saturday 25 November issue contains an entry of eleven pages entirely devoted to sugar. It discusses a broad range of topics related to sugar, including the role of sugar throughout the centuries, sugar consumption in the Netherlands, the amount of sugar in bread, and sugar

  4. Changes in sugar content and related enzyme activities in table grape (Vitis vinifera L.) in response to foliar selenium fertilizer.

    Science.gov (United States)

    Zhu, Shuaimeng; Liang, Yinli; An, Xiaojuan; Kong, Fanchao; Gao, Dekai; Yin, Hongfei

    2017-09-01

    Spraying selenium (Se) fertilizer is an effective method for Se-enriched fruit production. Sugar content in fruit is the major factor determining berry quality. However, changes in sugar metabolism in response to Se fertilizer are unclear. Hence, this study was conducted to identify the effects of Se fertilizer on sugar metabolism and related enzyme activities of grape berries. Additionally, production of leaves with and without Se fertilizer was also investigated. Acid invertase (AI) activity, total soluble sugar and Se content in berries, and photosynthetic rate in leaves produced under Se fertilizer treatments were higher than that of control. Glucose and fructose were the primary sugars in berries, with a trace of sucrose. In both berries and leaves, neutral invertase activity was lower than AI, there was no significant difference in neutral invertase, sucrose synthase and sucrose phosphate synthase between Se fertilizer-treated and control. In berries, AI showed a significant positive correlation with glucose and fructose; also Se content was significantly correlated with sugar content. AI played an important role in the process of sugar accumulation in berries; high AI activity in berries and photosynthetic rate in leaves could explain the mechanism by which Se fertilizer affected sugar accumulation in berries. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  5. Blood sugar test

    Science.gov (United States)

    ... sugar; Blood sugar level; Fasting blood sugar; Glucose test; Diabetic screening - blood sugar test; Diabetes - blood sugar test ... The test may be done in the following ways: After you have not eaten anything for at least 8 ...

  6. Indomethacin differentiates the renal effects of sphingosine-1-phosphate and sphingosylphosphorylcholine

    NARCIS (Netherlands)

    Czyborra, Claudia; Bischoff, Angela; Michel, Martin C.

    2006-01-01

    The sphingomyelin breakdown products sphingosine-1-phosphate (S1P) and sphingosylphosphorylcholine (SPC) constrict intrarenal microvessels in vitro in a pertussis toxin (PTX) sensitive manner, and S1P also reduces renal blood flow in vivo. Nevertheless, both S1P and SPC have been reported to enhance

  7. Lar mountain phosphate ore processing using flotation approach

    International Nuclear Information System (INIS)

    Gharabaghi, M.; Noaparast, M.; Shafaei Tonkaboni, S. Z.

    2007-01-01

    The sample of Lar Mountain Phosphate deposit which is located in the southwest of Iran, was studied to upgrade its phosphate grade. The results obtained from mineralogical studies showed the presence of apatite, CaO, Al 2 O 3 , Fe 2 O 3 and SiO 2 , in which carbonate was detected as the main gangue. Two sets of direct and reverse flotation tests were performed using samples from this deposit with 10% P 2 O 5 . In phosphate flotation (direct approach), the samples were conditioned with sodium silica, oleic acid-fuel oil and Armac T-fuel oil. The direct flotation at pH=9.2 yielded a product with 23.2% of P 2 O 5 and 75.16% recovery. The reverse flotation tests were carried out at pH=5.2, with floating carbonate and pulp de-oiling, using H 2 SO 4 and wash water, and phosphate was then floated from siliceous gangue. In the second sets of the reverse approach, depressing the phosphate and floating silica with Amines in natural pH were done. However the best concentrate assay was 31.2% P 2 O 5 with a 71.12% recovery, which was obtained from reverse tests

  8. Exposure to tri-o-cresyl phosphate detected in jet airplane passengers.

    Science.gov (United States)

    Liyasova, Mariya; Li, Bin; Schopfer, Lawrence M; Nachon, Florian; Masson, Patrick; Furlong, Clement E; Lockridge, Oksana

    2011-11-01

    The aircraft cabin and flight deck ventilation are supplied from partially compressed unfiltered bleed air directly from the engine. Worn or defective engine seals can result in the release of engine oil into the cabin air supply. Aircrew and passengers have complained of illness following such "fume events". Adverse health effects are hypothesized to result from exposure to tricresyl phosphate mixed esters, a chemical added to jet engine oil and hydraulic fluid for its anti-wear properties. Our goal was to develop a laboratory test for exposure to tricresyl phosphate. The assay was based on the fact that the active-site serine of butyrylcholinesterase reacts with the active metabolite of tri-o-cresyl phosphate, cresyl saligenin phosphate, to make a stable phosphorylated adduct with an added mass of 80 Da. No other organophosphorus agent makes this adduct in vivo on butyrylcholinesterase. Blood samples from jet airplane passengers were obtained 24-48 h after completing a flight. Butyrylcholinesterase was partially purified from 25 ml serum or plasma, digested with pepsin, enriched for phosphorylated peptides by binding to titanium oxide, and analyzed by mass spectrometry. Of 12 jet airplane passengers tested, 6 were positive for exposure to tri-o-cresyl phosphate that is, they had detectable amounts of the phosphorylated peptide FGEpSAGAAS. The level of exposure was very low. No more than 0.05 to 3% of plasma butyrylcholinesterase was modified. None of the subjects had toxic symptoms. Four of the positive subjects were retested 3 to 7 months following their last airplane trip and were found to be negative for phosphorylated butyrylcholinesterase. In conclusion, this is the first report of an assay that detects exposure to tri-o-cresyl phosphate in jet airplane travelers. Copyright © 2011 Elsevier Inc. All rights reserved.

  9. 76 FR 62339 - Domestic Sugar Program-2011-Crop Cane Sugar and Beet Sugar Marketing Allotments and Company...

    Science.gov (United States)

    2011-10-07

    ... DEPARTMENT OF AGRICULTURE Commodity Credit Corporation Domestic Sugar Program--2011-Crop Cane Sugar and Beet Sugar Marketing Allotments and Company Allocations AGENCY: Commodity Credit Corporation... the fiscal year (FY) 2012 State sugar marketing allotments and company allocations to sugarcane and...

  10. Sphingomyelinase D activity in model membranes: structural effects of in situ generation of ceramide-1-phosphate.

    Directory of Open Access Journals (Sweden)

    Roberto P Stock

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

  11. Molecular cloning of a novel glucuronokinase/putative pyrophosphorylase from zebrafish acting in an UDP-glucuronic acid salvage pathway.

    Directory of Open Access Journals (Sweden)

    Roman Gangl

    Full Text Available In animals, the main precursor for glycosaminoglycan and furthermore proteoglycan biosynthesis, like hyaluronic acid, is UDP-glucuronic acid, which is synthesized via the nucleotide sugar oxidation pathway. Mutations in this pathway cause severe developmental defects (deficiency in the initiation of heart valve formation. In plants, UDP-glucuronic acid is synthesized via two independent pathways. Beside the nucleotide sugar oxidation pathway, a second minor route to UDP-glucuronic acid exist termed the myo-inositol oxygenation pathway. Within this myo-inositol is ring cleaved into glucuronic acid, which is subsequently converted to UDP-glucuronic acid by glucuronokinase and UDP-sugar pyrophosphorylase. Here we report on a similar, but bifunctional enzyme from zebrafish (Danio rerio which has glucuronokinase/putative pyrophosphorylase activity. The enzyme can convert glucuronic acid into UDP-glucuronic acid, required for completion of the alternative pathway to UDP-glucuronic acid via myo-inositol and thus establishes a so far unknown second route to UDP-glucuronic acid in animals. Glucuronokinase from zebrafish is a member of the GHMP-kinase superfamily having unique substrate specificity for glucuronic acid with a Km of 31 ± 8 µM and accepting ATP as the only phosphate donor (Km: 59 ± 9 µM. UDP-glucuronic acid pyrophosphorylase from zebrafish has homology to bacterial nucleotidyltransferases and requires UTP as nucleosid diphosphate donor. Genes for bifunctional glucuronokinase and putative UDP-glucuronic acid pyrophosphorylase are conserved among some groups of lower animals, including fishes, frogs, tunicates, and polychaeta, but are absent from mammals. The existence of a second pathway for UDP-glucuronic acid biosynthesis in zebrafish likely explains some previous contradictory finding in jekyll/ugdh zebrafish developmental mutants, which showed residual glycosaminoglycans and proteoglycans in knockout mutants of UDP

  12. Consolidation of sugaring marble by hydroxyapatite: some recent developments on producing and treating decayed samples

    OpenAIRE

    Sassoni, Enrico; Graziani, Gabriela; Franzoni, Elisa; Scherer, George W.

    2016-01-01

    Consolidation of sugaring marble (i.e., marble affected by granular disaggregation) still lacks fully effective solutions. Consequently, the use of an innovative phosphate-based treatment, aimed at bonding calcite grains by formation of hydroxyapatite at grain boundaries, has recently been proposed. In this paper, firstly a novel method for producing artificially decayed marble samples, by contact with a heating plate, is proposed. Then, some results are presented about the effectiveness and ...

  13. A Nostoc punctiforme sugar transporter necessary to establish a Cyanobacterium-plant symbiosis.

    Science.gov (United States)

    Ekman, Martin; Picossi, Silvia; Campbell, Elsie L; Meeks, John C; Flores, Enrique

    2013-04-01

    In cyanobacteria-plant symbioses, the symbiotic nitrogen-fixing cyanobacterium has low photosynthetic activity and is supplemented by sugars provided by the plant partner. Which sugars and cyanobacterial sugar uptake mechanism(s) are involved in the symbiosis, however, is unknown. Mutants of the symbiotically competent, facultatively heterotrophic cyanobacterium Nostoc punctiforme were constructed bearing a neomycin resistance gene cassette replacing genes in a putative sugar transport gene cluster. Results of transport activity assays using (14)C-labeled fructose and glucose and tests of heterotrophic growth with these sugars enabled the identification of an ATP-binding cassette-type transporter for fructose (Frt), a major facilitator permease for glucose (GlcP), and a porin needed for the optimal uptake of both fructose and glucose. Analysis of green fluorescent protein fluorescence in strains of N. punctiforme bearing frt::gfp fusions showed high expression in vegetative cells and akinetes, variable expression in hormogonia, and no expression in heterocysts. The symbiotic efficiency of N. punctiforme sugar transport mutants was investigated by testing their ability to infect a nonvascular plant partner, the hornwort Anthoceros punctatus. Strains that were specifically unable to transport glucose did not infect the plant. These results imply a role for GlcP in establishing symbiosis under the conditions used in this work.

  14. Antioxidant and chelating capacity of Maillard reaction products in amino acid-sugar model systems: applications for food processing.

    Science.gov (United States)

    Mondaca-Navarro, Blanca A; Ávila-Villa, Luz A; González-Córdova, Aarón F; López-Cervantes, Jaime; Sánchez-Machado, Dalia I; Campas-Baypoli, Olga N; Rodríguez-Ramírez, Roberto

    2017-08-01

    Maillard reaction products (MRP) have gained increasing interest owing to their both positive and negative effects on human health. Aqueous amino acid-sugar model systems were studied in order to evaluate the antioxidant and chelating activity of MRP under conditions similar to those of food processing. Amino acids (cysteine, glycine, isoleucine and lysine) combined with different sugars (fructose or glucose) were heated to 100 and 130 °C for 30, 60 and 90 min. Antioxidant capacity was evaluated via ABTS and DPPH free radical scavenging assays, in addition to Fe 2+ and Cu 2+ ion chelating capacity. In the ABTS assay, the cysteine-fructose model system presented the highest antioxidant activity at 7.05 µmol mL -1 (130 °C, 60 min), expressed in Trolox equivalents. In the DPPH assay, the cysteine-glucose system presented the highest antioxidant activity at 3.79 µmol mL -1 (100 °C, 90 min). The maximum rate of chelation of Fe 2+ and Cu 2+ was 96.31 and 59.44% respectively in the lysine-fructose and cysteine-glucose systems (100 °C, 30 min). The model systems presented antioxidant and chelating activity under the analyzed temperatures and heating times, which are similar to the processing conditions of some foods. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  15. Structure of AadA from Salmonella enterica: a monomeric aminoglycoside (3′′)(9) adenyltransferase

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Yang [Uppsala University, Biomedical Center, Box 596, SE-751 24 Uppsala (Sweden); Näsvall, Joakim [Uppsala University, Biomedical Center, Box 582, SE-751 23 Uppsala (Sweden); Wu, Shiying [Uppsala University, Biomedical Center, Box 596, SE-751 24 Uppsala (Sweden); Andersson, Dan I. [Uppsala University, Biomedical Center, Box 582, SE-751 23 Uppsala (Sweden); Selmer, Maria, E-mail: maria.selmer@icm.uu.se [Uppsala University, Biomedical Center, Box 596, SE-751 24 Uppsala (Sweden)

    2015-10-31

    The crystal structure of the aminoglycoside-adenylating enzyme AadA is reported together with functional experiments providing insights into its oligomeric state, ligand binding and catalysis. Aminoglycoside resistance is commonly conferred by enzymatic modification of drugs by aminoglycoside-modifying enzymes such as aminoglycoside nucleotidyltransferases (ANTs). Here, the first crystal structure of an ANT(3′′)(9) adenyltransferase, AadA from Salmonella enterica, is presented. AadA catalyses the magnesium-dependent transfer of adenosine monophosphate from ATP to the two chemically dissimilar drugs streptomycin and spectinomycin. The structure was solved using selenium SAD phasing and refined to 2.5 Å resolution. AadA consists of a nucleotidyltransferase domain and an α-helical bundle domain. AadA crystallizes as a monomer and is a monomer in solution as confirmed by small-angle X-ray scattering, in contrast to structurally similar homodimeric adenylating enzymes such as kanamycin nucleotidyltransferase. Isothermal titration calorimetry experiments show that ATP binding has to occur before binding of the aminoglycoside substrate, and structure analysis suggests that ATP binding repositions the two domains for aminoglycoside binding in the interdomain cleft. Candidate residues for ligand binding and catalysis were subjected to site-directed mutagenesis. In vivo resistance and in vitro binding assays support the role of Glu87 as the catalytic base in adenylation, while Arg192 and Lys205 are shown to be critical for ATP binding.

  16. MC3T3-E1 proliferation and differentiation on biphasic mixtures of Mg substituted β-tricalcium phosphate and amorphous calcium phosphate

    International Nuclear Information System (INIS)

    Singh, Satish S.; Roy, Abhijit; Lee, Boeun E.; Banerjee, Ipsita; Kumta, Prashant N.

    2014-01-01

    A low temperature aqueous approach was used to synthesize nanocrystalline, high surface area Mg 2+ substituted β-tricalcium phosphate (β-TCMP) to assess its potential use as a synthetic bone graft substitute. X-ray diffraction indicated that β-TCMP was the predominant crystalline phase formed. However, thermal analysis revealed the presence of a secondary amorphous phase which increased with increasing Mg 2+ concentration. Further analysis by Rietveld refinement indicated that the level of ionic substitution of Ca 2+ by Mg 2+ was significantly lower than the amount of Mg 2+ measured using elemental analysis, confirming the formation of a Mg 2+ rich secondary amorphous phase. MC3T3-E1 proliferation on substrates prepared using β-TCMP was assessed using the MTT assay. In comparison to commercially available β-TCP, increased proliferation was observed on samples prepared with 50% Mg, despite elevated Mg 2+ and PO 4 3− concentrations in culture media. Alkaline phosphatase (ALP) activity and qRT-PCR were used to study the effect of varying Mg 2+ substitution on osteogenic differentiation. Cells cultured on β-TCMP substrates prepared with increased Mg 2+ concentrations expressed significantly increased levels of ALP activity and osteogenic genes such as, osteocalcin, collagen-1, and Runx2, in comparison to those cultured on commercially available β-TCP

  17. Radioactive waste package assay facility. Volume 1. Application of assay technology

    International Nuclear Information System (INIS)

    Findlay, D.J.S.; Green, T.H.; Molesworth, T.V.; Staniforth, D.; Strachan, N.R.; Rogers, J.D.; Wise, M.O.; Forrest, K.R.

    1992-01-01

    This report, in three volumes, covers the work carried out by Taylor Woodrow Construction Ltd., and two major sub-contractors: Harwell Laboratory (AEA Technology) and Siemens Plessey Controls Ltd., on the development of a radioactive waste package assay facility, for cemented 500 litre intermediate level waste drums. In volume 1, the reasons for assay are considered together with the various techniques that can be used, and the information that can be obtained. The practical problems associated with the use of the various techniques in an integrated assay facility are identified, and the key parameters defined. Engineering and operational features are examined and provisional designs proposed for facilities at three throughput levels: 15,000, 750 and 30 drums per year respectively. The capital and operating costs for such facilities have been estimated. A number of recommendations are made for further work. 16 refs., 14 figs., 13 tabs

  18. Preliminary crystallographic analysis of two hypothetical ribose-5-phosphate isomerases from Streptococcus mutans

    International Nuclear Information System (INIS)

    Wang, Chen; Fan, Xuexin; Cao, Xiaofang; Liu, Xiang; Li, Lanfen; Su, Xiaodong

    2012-01-01

    Two hypothetical ribose-5-phosphate isomerases from S. mutans have been produced in E. coli and crystallized. The crystals diffracted to high resolutions suitable for crystallographic analyses. Study of the enzymes from sugar metabolic pathways may provide a better understanding of the pathogenesis of the human oral pathogen Streptococcus mutans. Bioinformatics, biochemical and crystallization methods were used to characterize and understand the function of two putative ribose-5-phosphate isomerases: SMU1234 and SMU2142. The proteins were cloned and constructed with N-terminal His tags. Protein purification was performed by Ni 2+ -chelating and size-exclusion chromatography. The crystals of SUM1234 diffracted to 1.9 Å resolution and belonged to space group P2 1 2 1 2 1 , with unit-cell parameters a = 48.97, b = 98.27, c = 101.09 Å, α = β = γ = 90°. The optimized SMU2142 crystals diffracted to 2.7 Å resolution and belonged to space group P1, with unit-cell parameters a = 53.7, b = 54.1, c = 86.5 Å, α = 74.2, β = 73.5, γ = 83.7°. Initial phasing of both proteins was attempted by molecular replacement; the structure of SMU1234 could easily be solved, but no useful results were obtained for SMU2142. Therefore, SeMet-labelled SMU2142 will be prepared for phasing

  19. 27 CFR 24.181 - Use of sugar.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Use of sugar. 24.181... OF THE TREASURY LIQUORS WINE Production of Wine § 24.181 Use of sugar. Only sugar, as defined in § 24.10, may be used in the production of standard wine. The quantity of sugar used will be determined...

  20. Two MYB-related transcription factors play opposite roles in sugar signaling in Arabidopsis.

    Science.gov (United States)

    Chen, Yi-Shih; Chao, Yi-Chi; Tseng, Tzu-Wei; Huang, Chun-Kai; Lo, Pei-Ching; Lu, Chung-An

    2017-02-01

    Sugar regulation of gene expression has profound effects at all stages of the plant life cycle. Although regulation at the transcriptional level is one of the most prominent mechanisms by which gene expression is regulated, only a few transcription factors have been identified and demonstrated to be involved in the regulation of sugar-regulated gene expression. OsMYBS1, an R1/2-type MYB transcription factor, has been demonstrated to be involved in sugar- and hormone-regulated α-amylase gene expression in rice. Arabidopsis contains two OsMYBS1 homologs. In the present study, we investigate MYBS1 and MYBS2 in sugar signaling in Arabidopsis. Our results indicate that MYBS1 and MYBS2 play opposite roles in regulating glucose and ABA signaling in Arabidopsis during seed germination and early seedling development. MYB proteins have been classified into four subfamilies: R2R3-MYB, R1/2-MYB, 3R-MYB, and 4R-MYB. An R1/2-type MYB transcription factor, OsMYBS1, has been demonstrated to be involved in sugar- and hormone-regulated α-amylase genes expression in rice. In this study, two genes homologous to OsMYBS1, MYBS1 and MYBS2, were investigated in Arabidopsis. Subcellular localization analysis showed that MYBS1 and MYBS2 were localized in the nucleus. Rice embryo transient expression assays indicated that both MYBS1 and MYBS2 could recognize the sugar response element, TA-box, in the promoter and induced promoter activity. mybs1 mutant exhibited hypersensitivity to glucose, whereas mybs2 seedlings were hyposensitive to it. MYBS1 and MYBS2 are involved in the control of glucose-responsive gene expression, as the mybs1 mutant displayed increased expression of a hexokinase gene (HXK1), chlorophyll a/b-binding protein gene (CAB1), ADP-glucose pyrophosphorylase gene (APL3), and chalcone synthase gene (CHS), whereas the mybs2 mutant exhibited decreased expression of these genes. mybs1 also showed an enhanced response to abscisic acid (ABA) in the seed germination and seedling

  1. Lack of awareness among future medical professionals about the risk of consuming hidden phosphate-containing processed food and drinks.

    Directory of Open Access Journals (Sweden)

    Yoshiko Shutto

    Full Text Available Phosphate toxicity is an important determinant of mortality in patients with chronic kidney disease (CKD, particularly those undergoing hemodialysis treatments. CKD patients are advised to take a low phosphate-containing diet, and are additionally prescribed with phosphate-lowering drugs. Since these patients usually seek guidance from their physicians and nurses for their dietary options, we conducted a survey to determine the levels of awareness regarding the high phosphate content in commercially processed food and drinks among medical and nursing students at the Hirosaki University School of Medicine in Japan. For this survey, 190 medical and nursing students (average age 21.7±3 years were randomly selected, and provided with a list of questions aimed at evaluating their awareness of food and drinks containing artificially added phosphate ingredients. While 98.9% of these students were aware of the presence of sugar in commercially available soda drinks, only 6.9% were aware of the presence of phosphate (phosphoric acid. Similarly, only 11.6% of these students were aware of the presence of phosphate in commercially processed food, such as hamburgers and pizza. Moreover, around two thirds of the surveyed students (67.7% were unaware of the harmful effects of unrestricted consumption of phosphate-containing food and drinks. About 28% of the surveyed students consume such "fast food" once a week, while 40% drink at least 1∼5 cans of soda drinks/week. After realizing the potential long-term risks of consuming excessive phosphate-containing food and drinks, 40.5% of the survey participants considered reducing their phosphate intake by minimizing the consumption of commercially processed "fast food" items and soda drinks. Moreover, another 48.4% of students showed interest in obtaining more information on the negative health effects of consuming excessive amounts of phosphate. This survey emphasizes the need for educational initiative to raise

  2. Osh4p exchanges sterols for phosphatidylinositol 4-phosphate between lipid bilayers

    OpenAIRE

    de Saint-Jean, Maud; Delfosse, Vanessa; Douguet, Dominique; Chicanne, Gaetan; Payrastre, Bernard; Bourguet, William; Antonny, Bruno; Drin, Guillaume

    2011-01-01

    Osh/Orp proteins transport sterols between organelles and are involved in phosphoinositide metabolism. The link between these two aspects remains elusive. Using novel assays, we address the influence of membrane composition on the ability of Osh4p/Kes1p to extract, deliver, or transport dehydroergosterol (DHE). Surprisingly, phosphatidylinositol 4-phosphate (PI(4)P) specifically inhibited DHE extraction because PI(4)P was itself efficiently extracted by Osh4p. We solve the structure of the Os...

  3. Alpha 1A and alpha 1B-adrenoceptors enhance inositol phosphate generation in rat renal cortex

    NARCIS (Netherlands)

    Michel, M. C.; Büscher, R.; Philipp, T.; Brodde, O. E.

    1993-01-01

    We have studied the role of alpha 1A- and alpha 1B-adrenoceptors in noradrenaline- and methoxamine-stimulated inositol phosphate accumulation in rat renal cortical slices. [3H]Prazosin binding studies with and without inactivation of alpha 1B-adrenoceptors by chloroethylclonidine treatment suggested

  4. 27 CFR 24.317 - Sugar record.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Sugar record. 24.317... OF THE TREASURY LIQUORS WINE Records and Reports § 24.317 Sugar record. A proprietor who receives, stores, or uses sugar shall maintain a record of receipt and use. The record will show the date of...

  5. A new human NHERF1 mutation decreases renal phosphate transporter NPT2a expression by a PTH-independent mechanism.

    Directory of Open Access Journals (Sweden)

    Marie Courbebaisse

    Full Text Available BACKGROUND: The sodium-hydrogen exchanger regulatory factor 1 (NHERF1 binds to the main renal phosphate transporter NPT2a and to the parathyroid hormone (PTH receptor. We have recently identified mutations in NHERF1 that decrease renal phosphate reabsorption by increasing PTH-induced cAMP production in the renal proximal tubule. METHODS: We compared relevant parameters of phosphate homeostasis in a patient with a previously undescribed mutation in NHERF1 and in control subjects. We expressed the mutant NHERF1 protein in Xenopus Oocytes and in cultured cells to study its effects on phosphate transport and PTH-induced cAMP production. RESULTS: We identified in a patient with inappropriate renal phosphate reabsorption a previously unidentified mutation (E68A located in the PDZ1 domain of NHERF1.We report the consequences of this mutation on NHERF1 function. E68A mutation did not modify cAMP production in the patient. PTH-induced cAMP synthesis and PKC activity were not altered by E68A mutation in renal cells in culture. In contrast to wild-type NHERF1, expression of the E68A mutant in Xenopus oocytes and in human cells failed to increase phosphate transport. Pull down experiments showed that E68A mutant did not interact with NPT2a, which robustly interacted with wild type NHERF1 and previously identified mutants. Biotinylation studies revealed that E68A mutant was unable to increase cell surface expression of NPT2a. CONCLUSIONS: Our results indicate that the PDZ1 domain is critical for NHERF1-NPT2a interaction in humans and for the control of NPT2a expression at the plasma membrane. Thus we have identified a new mechanism of renal phosphate loss and shown that different mutations in NHERF1 can alter renal phosphate reabsorption via distinct mechanisms.

  6. Stability-Indicating HPLC Method for Simultaneous Determination of Chloramphenicol, Dexamethasone Sodium Phosphate and Tetrahydrozoline Hydrochloride in Ophthalmic Solution.

    Science.gov (United States)

    AlAani, Hashem; Alnukkary, Yasmin

    2016-03-01

    A simple stability-indicating RP-HPLC assay method was developed and validated for quantitative determination of Chloramphenicol, Dexamethasone Sodium Phosphate and Tetrahydrozoline Hydrochloride in ophthalmic solution in the presence of 2-amino-1-(4-nitrophenyl)propane-1,3-diol, a degradation product of Chloramphenicol, and Dexamethasone, a degradation product of Dexamethasone Sodium Phosphate. Effective chromatographic separation was achieved using C18 column (250 mm, 4.6 mm i.d., 5 μm) with isocratic mobile phase consisting of acetonitrile - phosphate buffer (pH 4.0; 0.05 M) (30:70, v/v) at a flow rate of 1 mL/minute. The column temperature was maintained at 40°C and the detection wavelength was 230 nm. The proposed HPLC procedure was statistically validated according to the ICH guideline, and was proved to be stability-indicating by resolution of the APIs from their forced degradation products. The developed method is suitable for the routine analysis as well as stability studies.

  7. Prolonging survival of corneal transplantation by selective sphingosine-1-phosphate receptor 1 agonist.

    Directory of Open Access Journals (Sweden)

    Min Gao

    Full Text Available Corneal transplantation is the most used therapy for eye disorders. Although the cornea is somewhat an immune privileged organ, immune rejection is still the major problem that reduces the success rate. Therefore, effective chemical drugs that regulate immunoreactions are needed to improve the outcome of corneal transplantations. Here, a sphingosine-1-phosphate receptor 1 (S1P1 selective agonist was systematically evaluated in mouse allogeneic corneal transplantation and compared with the commonly used immunosuppressive agents. Compared with CsA and the non-selective sphingosine 1-phosphate (S1P receptor agonist FTY720, the S1P1 selective agonist can prolong the survival corneal transplantation for more than 30 days with a low immune response. More importantly, the optimal dose of the S1P1 selective agonist was much less than non-selective S1P receptor agonist FTY720, which would reduce the dose-dependent toxicity in drug application. Then we analyzed the mechanisms of the selected S1P1 selective agonist on the immunosuppression. The results shown that the S1P1 selective agonist could regulate the distribution of the immune cells with less CD4+ T cells and enhanced Treg cells in the allograft, moreover the expression of anti-inflammatory cytokines TGF-β1 and IL-10 unregulated which can reduce the immunoreactions. These findings suggest that S1P1 selective agonist may be a more appropriate immunosuppressive compound to effectively prolong mouse allogeneic corneal grafts survival.

  8. Modulation of low dose radiation effect on pentose phosphate pathway enzymes by B-multivitamin deficiency

    International Nuclear Information System (INIS)

    Zimatkina, T.I.; Lashak, L.K.; Moiseenok, A.G.

    1997-01-01

    Blood, liver, thymus and spleen of albino rats injected subcutaneously with antivitamins (othythiamine and methotrexate) and subjected to prolonger γ-irradiation in the overall dose of 0.75 Gy were assayed for transketolase and glucose-6-phosphate dehydrogenase after 12h, 1, 2, 5 and 40 days from the last radiation dose. High transketolase sensitivity was found both to radiation (activation) and the combined effects of vitamin deficiency and radiation (potentiation of antivitamin inhibitory action) in all the tissues studied. The activity of glucose-6-phosphate dehydrogenase was little changed under the given experimental manipulations, but the combined effect of the factors considerably inhibited the enzyme activities in the organs of the immune system. Consequently, in B-multivitamin deficiency the effect of low radiation doses was subjected to a considerable modulation resulting in profound inhibition of the oxidation and nonoxidative branches of the pentose phosphate pathway. (author). 9 refs, 2 tabs

  9. The ultrasound-assisted sugar extraction from sugar beet cossettes

    International Nuclear Information System (INIS)

    Stasiak, D.M.

    2005-01-01

    The aim of this work was to study the ultrasound-assisted water extraction of sugar from sugar beet cossettes. The ultrasound bath device (25 kHz, 200 W) was used. The sonication accelerated sugar diffusion at both temperatures 18 deg C and 77.6 deg C and gave the higher level of dry matter content SS (4-6 percent) and sugar content CK (7-22 percent) in juice. The SS and CK depended on time of exposition, time and temperature of extraction. In particular, the effects of 5 min ultrasound-assisted extraction were equal to 20 min extraction in traditional conditions. The shorter time, lower temperature, higher efficiency and purity of juice could be the effects of sugar extraction with ultrasound. The change of thickness of diffusion membrane, microflows in tissue as well as it's environment caused by ultrasound was the reason of acceleration of sugar extraction

  10. HDL activation of endothelial sphingosine-1-phosphate receptor-1 (S1P1) promotes regeneration and suppresses fibrosis in the liver

    DEFF Research Database (Denmark)

    Ding, Bi-Sen; Liu, Catherine H; Sun, Yue

    2016-01-01

    Regeneration of hepatic sinusoidal vasculature is essential for non-fibrotic liver regrowth and restoration of its metabolic capacity. However, little is known about how this specialized vascular niche is regenerated. Here we show that activation of endothelial sphingosine-1-phosphate receptor-1 ...

  11. Sugars increase non-heme iron bioavailability in human epithelial intestinal and liver cells.

    Directory of Open Access Journals (Sweden)

    Tatiana Christides

    Full Text Available Previous studies have suggested that sugars enhance iron bioavailability, possibly through either chelation or altering the oxidation state of the metal, however, results have been inconclusive. Sugar intake in the last 20 years has increased dramatically, and iron status disorders are significant public health problems worldwide; therefore understanding the nutritional implications of iron-sugar interactions is particularly relevant. In this study we measured the effects of sugars on non-heme iron bioavailability in human intestinal Caco-2 cells and HepG2 hepatoma cells using ferritin formation as a surrogate marker for iron uptake. The effect of sugars on iron oxidation state was examined by measuring ferrous iron formation in different sugar-iron solutions with a ferrozine-based assay. Fructose significantly increased iron-induced ferritin formation in both Caco-2 and HepG2 cells. In addition, high-fructose corn syrup (HFCS-55 increased Caco-2 cell iron-induced ferritin; these effects were negated by the addition of either tannic acid or phytic acid. Fructose combined with FeCl3 increased ferrozine-chelatable ferrous iron levels by approximately 300%. In conclusion, fructose increases iron bioavailability in human intestinal Caco-2 and HepG2 cells. Given the large amount of simple and rapidly digestible sugars in the modern diet their effects on iron bioavailability may have important patho-physiological consequences. Further studies are warranted to characterize these interactions.

  12. Improved molecular tools for sugar cane biotechnology.

    Science.gov (United States)

    Kinkema, Mark; Geijskes, Jason; Delucca, Paulo; Palupe, Anthony; Shand, Kylie; Coleman, Heather D; Brinin, Anthony; Williams, Brett; Sainz, Manuel; Dale, James L

    2014-03-01

    Sugar cane is a major source of food and fuel worldwide. Biotechnology has the potential to improve economically-important traits in sugar cane as well as diversify sugar cane beyond traditional applications such as sucrose production. High levels of transgene expression are key to the success of improving crops through biotechnology. Here we describe new molecular tools that both expand and improve gene expression capabilities in sugar cane. We have identified promoters that can be used to drive high levels of gene expression in the leaf and stem of transgenic sugar cane. One of these promoters, derived from the Cestrum yellow leaf curling virus, drives levels of constitutive transgene expression that are significantly higher than those achieved by the historical benchmark maize polyubiquitin-1 (Zm-Ubi1) promoter. A second promoter, the maize phosphonenolpyruvate carboxylate promoter, was found to be a strong, leaf-preferred promoter that enables levels of expression comparable to Zm-Ubi1 in this organ. Transgene expression was increased approximately 50-fold by gene modification, which included optimising the codon usage of the coding sequence to better suit sugar cane. We also describe a novel dual transcriptional enhancer that increased gene expression from different promoters, boosting expression from Zm-Ubi1 over eightfold. These molecular tools will be extremely valuable for the improvement of sugar cane through biotechnology.

  13. Sphingosine 1-Phosphate and Atherosclerosis

    Science.gov (United States)

    Yatomi, Yutaka

    2018-01-01

    Sphingosine 1-phosphate (S1P) is a potent lipid mediator that works on five kinds of S1P receptors located on the cell membrane. In the circulation, S1P is distributed to HDL, followed by albumin. Since S1P and HDL share several bioactivities, S1P is believed to be responsible for the pleiotropic effects of HDL. Plasma S1P levels are reportedly lower in subjects with coronary artery disease, suggesting that S1P might be deeply involved in the pathogenesis of atherosclerosis. In basic experiments, however, S1P appears to possess both pro-atherosclerotic and anti-atherosclerotic properties; for example, S1P possesses anti-apoptosis, anti-inflammation, and vaso-relaxation properties and maintains the barrier function of endothelial cells, while S1P also promotes the egress and activation of lymphocytes and exhibits pro-thrombotic properties. Recently, the mechanism for the biased distribution of S1P on HDL has been elucidated; apolipoprotein M (apoM) carries S1P on HDL. ApoM is also a modulator of S1P, and the metabolism of apoM-containing lipoproteins largely affects the plasma S1P level. Moreover, apoM modulates the biological properties of S1P. S1P bound to albumin exerts both beneficial and harmful effects in the pathogenesis of atherosclerosis, while S1P bound to apoM strengthens anti-atherosclerotic properties and might weaken the pro-atherosclerotic properties of S1P. Although the detailed mechanisms remain to be elucidated, apoM and S1P might be novel targets for the alleviation of atherosclerotic diseases in the future. PMID:28724841

  14. Cross-sectional survey of the amount of free sugars and calories in carbonated sugar-sweetened beverages on sale in the UK

    Science.gov (United States)

    Hashem, Kawther M; He, Feng J; Jenner, Katharine H; MacGregor, Graham A

    2016-01-01

    Objectives To investigate the free sugars and calorie content of carbonated sugar-sweetened beverages (CSSB) available in the main UK supermarkets. Study design We carried out a cross-sectional survey in 2014 of 169 CSSB. Methods The free sugars (sugars g/100 mL) and calorie (kcal/100 mL) were collected from product packaging and nutrient information panels of CSSB available in 9 main UK supermarkets. Results The average free sugars content in CSSB was 30.1±10.7 g/330 mL, and 91% of CSSB would receive a ‘red’ (high) label for sugars per serving. There was a large variation in sugars content between different flavours of CSSB and within the same type of flavour ranging from 3.3 to 52.8 g/330 mL. On average, ginger beer (38.5±9.9 g/330 mL) contained the highest amounts of sugars and ginger ale (22.9±7.7 g/330 mL) contained the lowest. Cola flavour is the most popular flavour in the UK with an average free sugars content of 35.0±1.1 g/330 mL. On average, the supermarket own brand contained lower levels of sugars than branded products (27.9±10.6 vs 31.6±10.6 g/330 mL, p=0.02). The average calorie content in CSSB was 126.1±43.5 kcal/330 mL. Cola flavour had a calorie content of 143.5±5.2 kcal/330 mL. Among the 169 products surveyed, 55% exceeded the maximum daily recommendation for free sugars intake (30 g) per 330 mL. Conclusions Free sugars content of CSSB in the UK is high and is a major contributor to free sugars intake. There is a wide variation in the sugars content of CSSB and even within the same flavour of CSSB. These findings demonstrate that the amount of free sugars added to CSSB can be reduced without technical issues, and there is an urgent need to set incremental free sugars reduction targets. A reduction in sugars content and overall CSSB consumption will be very beneficial in reducing obesity, type 2 diabetes and dental caries. PMID:28186923

  15. Plasmodium falciparum infection increases Anopheles gambiae attraction to nectar sources and sugar uptake

    Science.gov (United States)

    Plasmodium parasites are known to manipulate the behaviour of their vectors so as to enhance their transmission. However, it is unknown if this vector manipulation also affects mosquito-plant interaction and sugar uptake. Dual-choice olfactometer and probing assays were used to study plant seeking b...

  16. Sugar beet breeding

    Science.gov (United States)

    Sugar beet is a recent crop developed solely for extraction of the sweetener sucrose. Breeding and improvement of Beta vulgaris for sugar has a rich historical record. Sugar beet originated from fodder beet in the 1800s, and selection has increased sugar content from 4 to 6% then to over 18% today. ...

  17. Glucose-6-phosphate reduces calcium accumulation in rat brain endoplasmic reticulum

    Directory of Open Access Journals (Sweden)

    Jeffrey Thomas Cole

    2012-04-01

    Full Text Available Brain cells expend large amounts of energy sequestering calcium (Ca2+, while loss of Ca2+ compartmentalization leads to cell damage or death. Upon cell entry, glucose is converted to glucose-6-phosphate (G6P, a parent substrate to several metabolic major pathways, including glycolysis. In several tissues, G6P alters the ability of the endoplasmic reticulum to sequester Ca2+. This led to the hypothesis that G6P regulates Ca2+ accumulation by acting as an endogenous ligand for sarco-endoplasmic reticulum calcium ATPase (SERCA. Whole brain ER microsomes were pooled from adult male Sprague-Dawley rats. Using radio-isotopic assays, 45Ca2+ accumulation was quantified following incubation with increasing amounts of G6P, in the presence or absence of thapsigargin, a potent SERCA inhibitor. To qualitatively assess SERCA activity, the simultaneous release of inorganic phosphate (Pi coupled with Ca2+ accumulation was quantified. Addition of G6P significantly and decreased Ca2+ accumulation in a dose-dependent fashion (1-10 mM. The reduction in Ca2+ accumulation was not significantly different that seen with addition of thapsigargin. Addition of glucose-1-phosphate or fructose-6-phosphate, or other glucose metabolic pathway intermediates, had no effect on Ca2+ accumulation. Further, the release of Pi was markedly decreased, indicating G6P-mediated SERCA inhibition as the responsible mechanism for reduced Ca2+ uptake. Simultaneous addition of thapsigargin and G6P did decrease inorganic phosphate in comparison to either treatment alone, which suggests that the two treatments have different mechanisms of action. Therefore, G6P may be a novel, endogenous regulator of SERCA activity. Additionally, pathological conditions observed during disease states that disrupt glucose homeostasis, may be attributable to Ca2+ dystasis caused by altered G6P regulation of SERCA activity

  18. Evaluation of synthase and hemisynthase activities of glucosamine-6-phosphate synthase by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Science.gov (United States)

    Gaucher-Wieczorek, Florence; Guérineau, Vincent; Touboul, David; Thétiot-Laurent, Sophie; Pelissier, Franck; Badet-Denisot, Marie-Ange; Badet, Bernard; Durand, Philippe

    2014-08-01

    Glucosamine-6-phosphate synthase (GlmS, EC 2.6.1.16) catalyzes the first and rate-limiting step in the hexosamine biosynthetic pathway, leading to the synthesis of uridine-5'-diphospho-N-acetyl-D-glucosamine, the major building block for the edification of peptidoglycan in bacteria, chitin in fungi, and glycoproteins in mammals. This bisubstrate enzyme converts D-fructose-6-phosphate (Fru-6P) and L-glutamine (Gln) into D-glucosamine-6-phosphate (GlcN-6P) and L-glutamate (Glu), respectively. We previously demonstrated that matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) allows determination of the kinetic parameters of the synthase activity. We propose here to refine the experimental protocol to quantify Glu and GlcN-6P, allowing determination of both hemisynthase and synthase parameters from a single assay kinetic experiment, while avoiding interferences encountered in other assays. It is the first time that MALDI-MS is used to survey the activity of a bisubstrate enzyme. Copyright © 2014 Elsevier Inc. All rights reserved.

  19. Increase The Sugar Concentration of The Solution Sugar by Reverse Osmotic Membrane

    Science.gov (United States)

    Redjeki, S.; Hapsari, N.; Iriani

    2018-01-01

    Sugar is one of the basic needs of people and food and drink industry. As technology advances and the demand for efficient usage of sugar rises, crystal sugar is seen as less advantageous than liquid sugar. If sugar is always dissolved in water before use, then it will be more efficient and practical for consumers to use sugar in liquid form than in crystal form. Other than that, liquid sugar is also attractive to consumers because it is economical, hygienic, instantly soluble in hot and cold water, fresher and longer-lasting, able to thicken and enrich the texture of foods and drinks, and functions as sweetener, syrup, and flavor enhancer. Liquid sugar is also more beneficial for sugar producers because of simpler production process, cheaper production cost, and similar yield with no extra cost. In sugar production, separation process is found in most of its stages and therefore the use of membrane technology for separating solute and water content has a good potential. In this research, water content reduction of sugar solution was done in order to increase the sugar concentration of the solution. The parameters of this research were 4%, 5%, and 6% starting concentration of sugar solution; 20, 40, and 60 minutes of process time; and 85 and 60 PSI ΔP. The best result was acquired on 4% starting concentration, 60 PSI ΔP, and 60 minutes process time.

  20. Improving conversion yield of fermentable sugars into fuel ethanol in 1st generation yeast-based production processes.

    Science.gov (United States)

    Gombert, Andreas K; van Maris, Antonius J A

    2015-06-01

    Current fuel ethanol production using yeasts and starch or sucrose-based feedstocks is referred to as 1st generation (1G) ethanol production. These processes are characterized by the high contribution of sugar prices to the final production costs, by high production volumes, and by low profit margins. In this context, small improvements in the ethanol yield on sugars have a large impact on process economy. Three types of strategies used to achieve this goal are discussed: engineering free-energy conservation, engineering redox-metabolism, and decreasing sugar losses in the process. Whereas the two former strategies lead to decreased biomass and/or glycerol formation, the latter requires increased process and/or yeast robustness. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. Further investigations on the inorganic phosphate binding site of beef heart mitochondrial F1-ATPase

    International Nuclear Information System (INIS)

    Pougeois, R.; Lauquin, G.J.

    1985-01-01

    The possibility that 4-azido-2-nitrophenyl phosphate (ANPP), a photoreactive derivative of inorganic phosphate (P /sub i/ ), could mimic ATP was investigated. ANPP was hydrolyzed in the dark by sarcoplasmic reticulum Ca 2+ -ATPase in the presence of Ca 2+ but not in the presence of ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid. ANPP was not hydrolyzed by purified mitochondrial F1-ATPase; however, ADP and ATP protected F1-ATPase against ANPP photoinactivation. On the other hand, the trinitrophenyl nucleotide analogues (TNP-ADP, TNP-ATP, and TNP-AMP-PNP), which bind specifically at the two catalytic sites of F1-ATPase, abolished P /sub i/ binding on F1-ATPase; they do not protect F1-ATPase against ANPP photoinactivation. Furthermore, ANPP-photoinactivated F1-ATPase binds the TNP analogues in the same way as the native enzyme. The Pi binding site of F1-ATPase, which is shown to be photolabeled by ANPP, does not appear to be at the gamma-phosphate position of the catalytic sites

  2. PROCESS OF OBTAINING OF SUGAR FROM SUGAR BEET AND INFLUENCE ON ITS QUALITY

    Directory of Open Access Journals (Sweden)

    Marián Tokár

    2011-02-01

    Full Text Available Cooking of massecuites has been study in the connection with different particle size distribution of white sugar. During the crystallization is possible to operate with parameters which have influence on particle size of white sugar. Dry matter of juice in crystallizer, volume of the standard syrup in crystallizer and heating curve of crystallization process are constant parameters in this process. Quantity of slurry (seed magma crystallizate and volume of slurry massecuite are parameters which can be changed for control the particle size distribution of white sugar. Five variants of viable parameters have been trying for obtain ideal particle size distribution of white sugar. As a best has been evaluated variant with 1100 cm3 of slurry and 20 % of volume of slurry massecuite in crystallizer. This variant has had the crystals proportions captured by the sieves between 1.00 and 0.40 mm with minimal differences in weight. More results have been related to reduction of losses of sugar in molasses with the right setting for the line of cooling crystallization process. The looses of sugar can be reduced by adding two coolers in the end of cooling crystalization process what will decrease a temperature to 40 ° C. This temperature will lead to more efficient crystallization in the cooling crystallization process.doi:10.5219/122

  3. Carbon quantum dots-based recyclable real-time fluorescence assay for alkaline phosphatase with adenosine triphosphate as substrate.

    Science.gov (United States)

    Qian, Zhaosheng; Chai, Lujing; Tang, Cong; Huang, Yuanyuan; Chen, Jianrong; Feng, Hui

    2015-03-03

    A convenient, reliable, and highly sensitive real-time assay for alkaline phosphatase (ALP) activity in the continuous and recyclable way is established on the basis of aggregation and disaggregation of carbon quantum dots (CQDs) through the competitive assay approach. CQDs and adenosine triphosphate (ATP) were used as the fluorescent indicator and substrate for ALP activity assessment, respectively. Richness of carboxyl groups on the surface of CQDs enables their severe aggregation triggered by cerium ions, which results in effective fluorescence quenching. Under the catalytic hydrolysis of ALP, ATP can be rapidly transformed to phosphate ions. Stronger affinity of phosphate ions to cerium ions than carboxyl groups is taken advantage of to achieve fluorescence recovery induced by redispersion of CQDs in the presence of ALP and ATP. Quantitative evaluation of ALP activity in a broad range from 4.6 to 383.3 U/L with the detection limit of 1.4 U/L can be realized in this way, which endows the assay with high enough sensitivity for practical detection in human serum. The assay can be used in a recyclable way for more than three times since the generated product CePO4 as a precipitate can be easily removed from the standard assay system. This strategy broadens the sensing application of fluorescent CQDs with excellent biocompatibility and provides an example based on disaggregation in optical probe development.

  4. Reactions of the melatonin metabolite N(1)-acetyl-5-methoxykynuramine with carbamoyl phosphate and related compounds.

    Science.gov (United States)

    Kuesel, Jana T; Hardeland, Rüdiger; Pfoertner, Henrike; Aeckerle, Nelia

    2010-01-01

    N-[2-(6-methoxyquinazolin-4-yl)-ethyl] acetamide (MQA) is a compound formed from the melatonin metabolite N(1)-acetyl-5-methoxykynuramine (AMK). We followed MQA production in reaction systems containing various putative reaction partners, in the absence and presence of hydrogen peroxide and/or copper(II). Although MQA may be formally described as a condensation product of either N(1)-acetyl-N(2)-formyl-5-methoxykynuramine (AFMK) with ammonia, or AMK with formamide, none of these combinations led to substantial quantities of MQA. However, MQA formation was observed in mixtures containing AMK, hydrogen peroxide, hydrogen carbonate and ammonia, or AMK, hydrogen peroxide, copper(II) and potentially carbamoylating agents, such as potassium cyanate or, more efficiently, carbamoyl phosphate. In the presence of hydrogen peroxide, copper(II) and carbamoyl phosphate, MQA was the major product obtained from AMK, but the omission of copper(II) mainly led to another metabolite, 3-acetamidomethyl-6-methoxycinnolinone (AMMC). This was caused by nitric oxide (NO) generated under oxidative conditions from carbamoyl phosphate, as shown by an NO spin trap. MQA formation with carbamoyl phosphate was not due to the possible decomposition product, formamide. The reaction of AMK with carbamoyl phosphate under oxidative conditions, in which inorganic phosphate and water are released and which differs from the typical process of carbamoylation via isocyanate, may be considered as a new physiological route of MQA formation.

  5. The effect of honey compared to sucrose, mixed sugars, and a sugar-free diet on weight gain in young rats.

    Science.gov (United States)

    Chepulis, L M

    2007-04-01

    To determine whether honey, sucrose, and mixed sugars as in honey have different effects on weight gain, 40 6-wk-old Sprague-Dawley rats were fed a powdered diet that was either sugar free or contained 8% sucrose, 8% mixed sugars as in honey, or 10% honey freely for 6 wk. Weight gain and food intake were assessed weekly, and at completion of the study blood samples were removed for measurement of blood sugar (HbA1c) and a fasting lipid profile. The animals were then minced and total percentage body fat and protein measured. Overall percentage weight gain was significantly lower in honey-fed rats than those fed sucrose or mixed sugars, despite a similar food intake. Weight gains were comparable for rats fed honey and a sugar free diet although food intake was significantly higher in honey-fed rats. HbA1c and triglyceride levels were significantly higher in all sugar treatments compared with rats fed a sugar free diet, but no other differences in lipid profiles were reported. No differences in percentage body fat or protein levels were reported.

  6. Sustainability Issues and Opportunities in the Sugar and Sugar-Bioproduct Industries

    Directory of Open Access Journals (Sweden)

    Gillian Eggleston

    2015-09-01

    Full Text Available Like many other industries, the sugar and sugar-bioproduct industries are facing important sustainability issues and opportunities. The relatively low and fluctuating profit for sugar, surpluses of sugar, world-wide trend to produce alternative, renewable bio-based fuels and chemicals to those derived from petroleum and reduce greenhouse gases, water- and energy-intensive factories and refineries, and increased consumer demands for sustainably manufactured products are putting pressure on the industries to diversify for sustainability. Sugar crops, including sugar and energy cane (Saccharum officinarum, sugar and energy beets (Beta vulgaris, and sweet sorghum (Sorghum bicolor L. Moench, are excellent, renewable biomass feedstocks because of their availability, their being amongst the plants that give the highest yields of carbohydrates per hectare, and high sugar contents. While much research has been focused on conversion technologies for advanced biofuels and bioproducts, attention is now focused on developing sustainable supply chains of sugar feedstocks for the new, flexible biorefineries, with customers wanting maximum feedstock reliability and quality, while minimizing cost. All biomass from sugar crops are potential feedstocks. The cogeneration of bioelectricity from bagasse and leaf residues is being increasingly manufactured in more countries and, due to the high carbon content of bagasse and leaves, can also be converted into value-added products such as biochar. Sugar crops are superior feedstocks for the production of platform chemicals for the manufacture of a range of end-products, e.g., bioplastics, chemicals, and biomaterials. In several countries and regions, green sustainability criteria are now in place and have to be met to count against national biofuel targets. Processes to convert high-fiber sugar crop biomass into biofuel have been developed but there has only been limited commercialization at the large-scale.

  7. Role of potassium and nitrogen on sugar concentration of sugar beet

    African Journals Online (AJOL)

    Sugar is obtained from root of sugar beet (Beta vulgaris L.) in addition to other sources. Three important economic parameters are often considered and these are root yield, sugar concentration in root juice and total sugar yield. All the three are affected by cropping period and use of fertilisers. Existing literature suggests the ...

  8. Oral antibodies to human intestinal alkaline phosphatase reduce dietary phytate phosphate bioavailability in the presence of dietary 1α-hydroxycholecalciferol.

    Science.gov (United States)

    Bobeck, Elizabeth A; Hellestad, Erica M; Helvig, Christian F; Petkovich, P Martin; Cook, Mark E

    2016-03-01

    While it is well established that active vitamin D treatment increases dietary phytate phosphate utilization, the mechanism by which intestinal alkaline phosphatase (IAP) participates in phytate phosphate use is less clear. The ability of human IAP (hIAP) oral antibodies to prevent dietary phytate phosphate utilization in the presence of 1α-hydroxycholecalciferol (1α-(OH) D3) in a chick model was investigated. hIAP specific chicken immunoglobulin Y (IgY) antibodies were generated by inoculating laying hens with 17 synthetic peptides derived from the human IAP amino acid sequence and harvesting egg yolk. Western blot analysis showed all antibodies recognized hIAP and 6 of the 8 antibodies selected showed modest inhibition of hIAP activity in vitro (6 to 33% inhibition). In chicks where dietary phosphate was primarily in the form of phytate, 4 selected hIAP antibodies inhibited 1α-(OH) D3-induced increases in blood phosphate, one of which, generated against selected peptide (MFPMGTPD), was as effective as sevelamer hydrochloride in preventing the 1α-(OH) D3-induced increase in blood phosphate, but ineffective in preventing an increase in body weight gain and bone ash induced by 1α-(OH) D3. These studies demonstrated that orally-delivered antibodies to IAP limit dietary phytate-phosphate utilization in chicks treated with 1α-(OH) D3, and implicate IAP as an important host enzyme in increasing phytate phosphate bioavailability in 1α-(OH) D3 fed chicks. © 2015 Poultry Science Association Inc.

  9. Phosphate steering by Flap Endonuclease 1 promotes 5′-flap specificity and incision to prevent genome instability

    KAUST Repository

    Tsutakawa, Susan E.; Thompson, Mark J.; Arvai, Andrew S.; Neil, Alexander J.; Shaw, Steven J.; Algasaier, Sana I.; Kim, Jane C.; Finger, L. David; Jardine, Emma; Gotham, Victoria J.B.; Sarker, Altaf H.; Her, Mai Z.; Rashid, Fahad; Hamdan, Samir; Mirkin, Sergei M.; Grasby, Jane A.; Tainer, John A.

    2017-01-01

    and large-scale trinucleotide (GAA) repeat expansions in vivo, implying failed phosphate-steering promotes an unanticipated lagging-strand template-switch mechanism during replication. Thus, phosphate steering is an unappreciated FEN1 function that enforces

  10. Photometric Determination of Ammonium and Phosphate in Seawater Medium Using a Microplate Reader.

    Science.gov (United States)

    Ruppersberg, Hanna S; Goebel, Maren R; Kleinert, Svea I; Wünsch, Daniel; Trautwein, Kathleen; Rabus, Ralf

    2017-01-01

    To more efficiently process the large sample numbers for quantitative determination of ammonium (NH4+) and phosphate (orthophosphate, PO43-) generated during comprehensive growth experiments with the marine Roseobacter group member Phaeobacter inhibens DSM 17395, specific colorimetric assays employing a microplate reader (MPR) were established. The NH4+ assay is based on the reaction of NH4+ with hypochlorite and salicylate, yielding a limit of detection of 14 µM, a limit of quantitation of 36 µM, and a linear range for quantitative determination up to 200 µM. The PO43-assay is based on the complex formation of PO43- with ammonium molybdate in the presence of ascorbate and zinc acetate, yielding a limit of detection of 13 µM, a limit of quantitation of 50 µM, and a linear range for quantitative determination up to 1 mM. Both MPR-based assays allowed for fast (significantly lower than 1 h) analysis of 21 samples plus standards for calibration (all measured in triplicates) and showed only low variation across a large collection of biological samples. © 2017 S. Karger AG, Basel.

  11. AN ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) METHOD FOR THE URINARY BIOMONITORING OF 2,4-DICHLOROPHRENOCYACETIC ACID (2,4-D)

    Science.gov (United States)

    An enzyme-linked immunosorbent assay (ELISA) method was developed to quantitatively measure 2,4-dichlorophenoyacetic acid (2,4-D) in human urine. Samples were diluted (1:5) with phosphate-buffered saline, 0.05% Tween 20, with 0.02% sodium azide, and analyzed by a 96-microwekk pl...

  12. Total, added, and free sugars: are restrictive guidelines science-based or achievable?

    Science.gov (United States)

    Erickson, Jennifer; Slavin, Joanne

    2015-04-15

    Sugar consumption, especially added sugars, is under attack. Various government and health authorities have suggested new sugar recommendations and guidelines as low as 5% of total calories from free sugars. Definitions for total sugars, free sugars, and added sugars are not standardized, nor are there accepted nutrient databases for this information. Our objective was to measure total sugars and added sugars in sample meal plans created by the United States Department of Agriculture (USDA) and the Academy of Nutrition and Dietetics (AND). Utilizing the Nutrition Data System for Research (NDSR) nutritional database, results found that plans created by the USDA and AND averaged 5.1% and 3.1% calories from added sugar, 8.7% and 3.1% from free sugar, and 23.3% and 21.1% as total sugars respectively. Compliance with proposed added sugar recommendations would require strict dietary compliance and may not be sustainable for many Americans. Without an accepted definition and equation for calculating added sugar, added sugar recommendations are arbitrary and may reduce intakes of nutrient-rich, recommended foods, such as yogurt, whole grains, and tart fruits including cranberries, cherries, and grapefruit. Added sugars are one part of excess calorie intake; however, compliance with low added sugar recommendations may not be achievable for the general public.

  13. Total, Added, and Free Sugars: Are Restrictive Guidelines Science-Based or Achievable?

    Directory of Open Access Journals (Sweden)

    Jennifer Erickson

    2015-04-01

    Full Text Available Sugar consumption, especially added sugars, is under attack. Various government and health authorities have suggested new sugar recommendations and guidelines as low as 5% of total calories from free sugars. Definitions for total sugars, free sugars, and added sugars are not standardized, nor are there accepted nutrient databases for this information. Our objective was to measure total sugars and added sugars in sample meal plans created by the United States Department of Agriculture (USDA and the Academy of Nutrition and Dietetics (AND. Utilizing the Nutrition Data System for Research (NDSR nutritional database, results found that plans created by the USDA and AND averaged 5.1% and 3.1% calories from added sugar, 8.7% and 3.1% from free sugar, and 23.3% and 21.1% as total sugars respectively. Compliance with proposed added sugar recommendations would require strict dietary compliance and may not be sustainable for many Americans. Without an accepted definition and equation for calculating added sugar, added sugar recommendations are arbitrary and may reduce intakes of nutrient-rich, recommended foods, such as yogurt, whole grains, and tart fruits including cranberries, cherries, and grapefruit. Added sugars are one part of excess calorie intake; however, compliance with low added sugar recommendations may not be achievable for the general public.

  14. Managing your blood sugar

    Science.gov (United States)

    Hyperglycemia - control; Hypoglycemia - control; Diabetes - blood sugar control; Blood glucose - managing ... sugar ( hypoglycemia ) Recognize and treat high blood sugar ( hyperglycemia ) Plan healthy meals Monitor your blood sugar (glucose) ...

  15. Synthesis of tritium labelled phosphonate analogues of sphinganine-1-phosphate

    International Nuclear Information System (INIS)

    Schick, Andreas; Schwarzmann, Guenter; Kolter, Thomas; Sandhoff, Konrad

    1997-01-01

    Tritiated phosphonate analogues 9 and 10 are prepared as analogues of sphinganine-1-phosphate 4. The key step in this synthesis is the catalytic tritiation of the triple bond in reduction of the protected diethyl-3-(S)-tert.-butoxycarbonylamino -4-hydroxy-5-tridecinyl-1-phosphonate by means of sodium boro[ 3 H]hydride as tritium source. These compounds are synthesized to study their metabolic stability and to evaluate their biological properties. (author)

  16. Method of stripping plutonium from tributyl phosphate solution which contains dibutyl phosphate-plutonium stable complexes

    International Nuclear Information System (INIS)

    Ochsenfeld, W.; Schmieder, H.

    1976-01-01

    Fast breeder fuel elements which have been highly burnt-up are reprocessed by extracting uranium and plutonium into an organic solution containing tributyl phosphate. The tributyl phosphate degenerates at least partially into dibutyl phosphate and monobutyl phosphate, which form stable complexes with tetravalent plutonium in the organic solution. This tetravalent plutonium is released from its complexed state and stripped into aqueous phase by contacting the organic solution with an aqueous phase containing tetravalent uranium. 6 claims, 1 drawing figure

  17. A regenerable potassium and phosphate sorbent system to enhance dialysis efficacy and device portability: an in vitro study.

    Science.gov (United States)

    Wester, Maarten; Simonis, Frank; Gerritsen, Karin G; Boer, Walther H; Wodzig, Will K; Kooman, Jeroen P; Joles, Jaap A

    2013-09-01

    Continuous dialysis could provide benefit by constant removal of potassium and phosphate. This study investigates the suitability of specific potassium and phosphate sorbents for incorporation in an extracorporeal device by capacity and regenerability testing. Capacity testing was performed in uraemic plasma. Regenerability was tested for potassium sorbents, with adsorption based on cationic exchange for sodium, with 0.1 M and 1.0 M NaCl. To regenerate phosphate sorbents, with adsorption based on anionic exchange, 0.1 M and 1.0 M NaHCO3 and NaOH were used. Subsequently, sodium polystyrene divinylbenzene sulphonate (RES-A) and iron oxide hydroxide (FeOOH) beads were incorporated in a cartridge for testing in bovine blood using a recirculating blood circuit and a dialysis circuit separated by a high-flux dialyzer (dynamic setup). Preloading was tested to assess whether this could limit calcium and magnesium adsorption. In the batch-binding assays, zirconium phosphate most potently adsorbed potassium (0.44 ± 0.05 mmol/g) and RES-A was the best regenerable potassium sorbent (92.9 ± 5.7% with 0.1 M NaCl). Zirconium oxide hydroxide (ZIR-hydr) most potently adsorbed phosphate (0.23 ± 0.05 mmol/g) and the polymeric amine sevelamer carbonate was the best regenerable sorbent (85.7 ± 5.2% with 0.1 M NaHCO3). In the dynamic setup, a potassium adsorption of 10.72 ± 2.06 mmol in 3 h was achieved using 111 g of RES-A and a phosphate adsorption of 4.73 ± 0.53 mmol in 3 h using 55 g of FeOOH. Calcium and magnesium preloading was shown to reduce the net adsorption in 3 h from 3.57 ± 0.91 to -0.29 ± 1.85 and 1.02 ± 0.05 to -0.31 ± 0.18 mmol, respectively. RES-A and FeOOH are suitable, regenerizable sorbents for potassium and phosphate removal in dialysate regeneration. Use of zirconium carbonate and ZIR-hydr may further increase phosphate adsorption, but may compromise sorbent regenerability. Use of polymeric amines for phosphate adsorption may enhance sorbent

  18. STUDY ON THE SUGAR-ACID RATIO AND RELEVANT METABOLIZING ENZYME ACTIVITIES IN NAVEL ORANGE FRUITS FROM DIFFERENT ECO-REGIONS

    Directory of Open Access Journals (Sweden)

    GONG RONGGAO

    2015-12-01

    Full Text Available ABSTRACT The flavor quality of citrus fruits is largely determined by the sugar-acid ratio, but it remains uncertain how sugar- and/or acid-metabolizing enzymes regulate the sugar-acid ratio of navel oranges and further affect the fruit quality. In the present study, Robertson navel oranges (Citrus sinesis Osb. were collected from six representative habitats in three eco-regions of Sichuan, China. The changes in the sugar-acid ratio and the activities of sucrose phosphate synthase (SPS, sucrose synthase (SS, cytosolic cio-aconitase (ACO, and isocitrate dehydrogenase (IDH were examined in navel oranges during fruit development. The results indicated that the sugar-acid ratio of fruits in different eco-regions changed significantly from 150 days after full bloom. The SPS and cytosolic ACO fruit activities had minor changes among different ecoregions throughout the experimental periods, whereas the activities of SS and IDH changed significantly in fruits among three eco-regions. Furthermore, the sugar-acid ratio and the activities of SS in the synthetic direction and IDH were the highest in south subtropics and the lowest in north mid-subtropics, probably due to the effects of climate conditions and/or other relevant eco-factors. It demonstrated that SS in the synthetic direction and IDH were of greater importance in regulating the sugar-acid ratio of navel oranges in different eco-regions, which provided new insights into the factors that determine the flavor quality of navel oranges and valuable data for guiding relevant agricultural practices.

  19. Arginine-phosphate salt bridges between histones and DNA: Intermolecular actuators that control nucleosome architecture

    Science.gov (United States)

    Yusufaly, Tahir I.; Li, Yun; Singh, Gautam; Olson, Wilma K.

    2014-10-01

    Structural bioinformatics and van der Waals density functional theory are combined to investigate the mechanochemical impact of a major class of histone-DNA interactions, namely, the formation of salt bridges between arginine residues in histones and phosphate groups on the DNA backbone. Principal component analysis reveals that the configurational fluctuations of the sugar-phosphate backbone display sequence-specific directionality and variability, and clustering of nucleosome crystal structures identifies two major salt-bridge configurations: a monodentate form in which the arginine end-group guanidinium only forms one hydrogen bond with the phosphate, and a bidentate form in which it forms two. Density functional theory calculations highlight that the combination of sequence, denticity, and salt-bridge positioning enables the histones to apply a tunable mechanochemical stress to the DNA via precise and specific activation of backbone deformations. The results suggest that selection for specific placements of van der Waals contacts, with high-precision control of the spatial distribution of intermolecular forces, may serve as an underlying evolutionary design principle for the structure and function of nucleosomes, a conjecture that is corroborated by previous experimental studies.

  20. Combination of sugar analysis and stable isotope ratio mass spectrometry to detect the use of artificial sugars in royal jelly production.

    Science.gov (United States)

    Wytrychowski, Marine; Daniele, Gaëlle; Casabianca, Hervé

    2012-05-01

    The effects of feeding bees artificial sugars and/or proteins on the sugar compositions and (13)C isotopic measurements of royal jellies (RJs) were evaluated. The sugars fed to the bees were two C4 sugars (cane sugar and maize hydrolysate), two C3 sugars (sugar beet, cereal starch hydrolysate), and honey. The proteins fed to them were pollen, soybean, and yeast powder proteins. To evaluate the influence of the sugar and/or protein feeding over time, samples were collected during six consecutive harvests. (13)C isotopic ratio measurements of natural RJs gave values of around -25 ‰, which were also seen for RJs obtained when the bees were fed honey or C3 sugars. However, the RJs obtained when the bees were fed cane sugar or corn hydrolysate (regardless of whether they were also fed proteins) gave values of up to -17 ‰. Sugar content analysis revealed that the composition of maltose, maltotriose, sucrose, and erlose varied significantly over time in accordance with the composition of the syrup fed to the bees. When corn and cereal starch hydrolysates were fed to the bees, the maltose and maltotriose contents of the RJs increased up to 5.0 and 1.3 %, respectively, compared to the levels seen in authentic samples (i.e., samples obtained when the bees were fed natural food: honey and pollen) that were inferior to 0.2% and not detected, respectively. The sucrose and erlose contents of natural RJs were around 0.2 %, whereas those in RJs obtained when the bees were fed cane or beet sugar were as much as 4.0 and 1.3 %, respectively. The combination of sugar analysis and (13)C isotopic ratio measurements represents a very efficient analytical methodology for detecting (from early harvests onward) the use of C4 and C3 artificial sugars in the production of RJ.

  1. Environmentally relevant organophosphate triesters in herring gulls: In vitro biotransformation and kinetics and diester metabolite formation using a hepatic microsomal assay

    International Nuclear Information System (INIS)

    Greaves, Alana K.; Su, Guanyong; Letcher, Robert J.

    2016-01-01

    The in vitro biotransformation and kinetics of six organophosphate triester (OPE) flame retardants were investigated in herring gulls (Larus argentatus) from the Great Lakes using a hepatic microsomal metabolism assay. Administration of each individual OPE (tri-n-butyl phosphate (TNBP), tris(2-butoxyethyl) phosphate (TBOEP), triphenyl phosphate (TPHP), triethyl phosphate (TEP), tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) and tris(2-chloroisopropyl) phosphate (TCIPP)) to the in vitro assay (concentration range 0.01 to 10 μM) resulted in rapid depletion with the exception of TEP. Following the Michaelis-Menten enzyme kinetics model, a preliminary 2-minute incubation period was used to estimate the V max (± SE) values (i.e., the maximal rate of reaction for a saturated enzyme system), which ranged from 5.0 ± 0.4 (TPHP) to 29 ± 18 pmol/min/mg protein (TBOEP), as well as the K M (± SE) values (i.e., the OPE concentration corresponding to one half of the V max ), which ranged from 9.8 ± 1 (TPHP) to 189 ± 135 nM (TBOEP). Biotransformation assays over a 100-minute incubation period revealed that TNBP was metabolized most rapidly (with a depletion rate of 73 ± 4 pmol/min/mg protein), followed by TBOEP (53 ± 8 pmol/min/mg), TCIPP (27 ± 1 pmol/min/mg), TPHP (22 ± 2 pmol/min/mg) and TDCIPP (8 ± 1 pmol/min/mg). In vitro biotransformation of OP triesters was clearly structure-dependent where non-halogenated alkyl OP triesters were metabolized more rapidly than halogenated alkyl triesters. Halogenated OP triesters were transformed to their respective diesters more efficiently relative to non-halogenated OP triesters. To our knowledge, this is the first study to investigate OP triester metabolism and OP diester formation in an avian or wildlife model system, which is important to understand the fate and biological activity of OPEs in an exposed organism. - Highlights: • The metabolism and kinetics of 6 OPEs were examined in herring gull liver microsomes. • The

  2. Environmentally relevant organophosphate triesters in herring gulls: In vitro biotransformation and kinetics and diester metabolite formation using a hepatic microsomal assay

    Energy Technology Data Exchange (ETDEWEB)

    Greaves, Alana K. [Wildlife and Landscape Directorate, Science and Technology Branch, Environment and Climate Change Canada, National Wildlife Research Centre, Carleton University, Ottawa, ON K1A 0H3 (Canada); Department of Chemistry, Carleton University, Ottawa, ON K1S 5B6 (Canada); Su, Guanyong, E-mail: guanyong.su85@gmail.com [Wildlife and Landscape Directorate, Science and Technology Branch, Environment and Climate Change Canada, National Wildlife Research Centre, Carleton University, Ottawa, ON K1A 0H3 (Canada); Department of Chemistry, Carleton University, Ottawa, ON K1S 5B6 (Canada); Letcher, Robert J., E-mail: robert.letcher@canada.ca [Wildlife and Landscape Directorate, Science and Technology Branch, Environment and Climate Change Canada, National Wildlife Research Centre, Carleton University, Ottawa, ON K1A 0H3 (Canada); Department of Chemistry, Carleton University, Ottawa, ON K1S 5B6 (Canada)

    2016-10-01

    The in vitro biotransformation and kinetics of six organophosphate triester (OPE) flame retardants were investigated in herring gulls (Larus argentatus) from the Great Lakes using a hepatic microsomal metabolism assay. Administration of each individual OPE (tri-n-butyl phosphate (TNBP), tris(2-butoxyethyl) phosphate (TBOEP), triphenyl phosphate (TPHP), triethyl phosphate (TEP), tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) and tris(2-chloroisopropyl) phosphate (TCIPP)) to the in vitro assay (concentration range 0.01 to 10 μM) resulted in rapid depletion with the exception of TEP. Following the Michaelis-Menten enzyme kinetics model, a preliminary 2-minute incubation period was used to estimate the V{sub max} (± SE) values (i.e., the maximal rate of reaction for a saturated enzyme system), which ranged from 5.0 ± 0.4 (TPHP) to 29 ± 18 pmol/min/mg protein (TBOEP), as well as the K{sub M} (± SE) values (i.e., the OPE concentration corresponding to one half of the V{sub max}), which ranged from 9.8 ± 1 (TPHP) to 189 ± 135 nM (TBOEP). Biotransformation assays over a 100-minute incubation period revealed that TNBP was metabolized most rapidly (with a depletion rate of 73 ± 4 pmol/min/mg protein), followed by TBOEP (53 ± 8 pmol/min/mg), TCIPP (27 ± 1 pmol/min/mg), TPHP (22 ± 2 pmol/min/mg) and TDCIPP (8 ± 1 pmol/min/mg). In vitro biotransformation of OP triesters was clearly structure-dependent where non-halogenated alkyl OP triesters were metabolized more rapidly than halogenated alkyl triesters. Halogenated OP triesters were transformed to their respective diesters more efficiently relative to non-halogenated OP triesters. To our knowledge, this is the first study to investigate OP triester metabolism and OP diester formation in an avian or wildlife model system, which is important to understand the fate and biological activity of OPEs in an exposed organism. - Highlights: • The metabolism and kinetics of 6 OPEs were examined in herring gull liver

  3. Phosphate Starvation Inducible Metabolism in Lycopersicon esculentum1

    Science.gov (United States)

    Goldstein, Alan H.; Baertlein, Dawn A.; McDaniel, Robert G.

    1988-01-01

    Both tomato (Lycopersicon esculentum cv VF 36) plants and suspension cultured cells show phosphate starvation inducible (psi) excretion of acid phosphatase (Apase). Apase excretion in vitro was proportional to the level of exogenous orthophosphate (Pi). Intracellular Apase activity remained the same in both Pi-starved and sufficient cells, while Apase excreted by the starved cells increased by as much as six times over unstressed control cells on a dry weight basis. At peak induction, 50% of total Apase was excreted. Ten day old tomato seedlings grown without Pi showed slight growth reduction versus unstressed control plants. The Pi-depleted roots showed psi enhancement of Apase activity. Severely starved seedlings (17 days) reached only one-third of the biomass of unstressed control plants but, because of a combination of psi Apase excretion by roots and a shift in biomass to this organ, they excreted 5.5 times the Apase activity of the unstressed control. Observed psi Apase excretion may be part of a phosphate starvation rescue system in plants. The utility of the visible indicator dye 5-bromo-4-chloro-3-indolyl-phosphate-p-toluidine as a phenotypic marker for plant Apase excretion is demonstrated. Images Fig. 5 PMID:16666212

  4. Association of Sphingosine-1-phosphate (S1P)/S1P Receptor-1 Pathway with Cell Proliferation and Survival in Canine Hemangiosarcoma.

    Science.gov (United States)

    Rodriguez, A M; Graef, A J; LeVine, D N; Cohen, I R; Modiano, J F; Kim, J-H

    2015-01-01

    Sphingosine-1-phosphate (S1P) is a key biolipid signaling molecule that regulates cell growth and survival, but it has not been studied in tumors from dogs. S1P/S1P1 signaling will contribute to the progression of hemangiosarcoma (HSA). Thirteen spontaneous HSA tissues, 9 HSA cell lines, 8 nonmalignant tissues, including 6 splenic hematomas and 2 livers with vacuolar degeneration, and 1 endothelial cell line derived from a dog with splenic hematoma were used. This was a retrospective case series and in vitro study. Samples were obtained as part of medically necessary diagnostic procedures. Microarray, qRT-PCR, immunohistochemistry, and immunoblotting were performed to examine S1P1 expression. S1P concentrations were measured by high-performance liquid chromatography/mass spectrometry. S1P signaling was evaluated by intracellular Ca(2+) mobilization; proliferation and survival were evaluated using the MTS assay and Annexin V staining. Canine HSA cells expressed higher levels of S1P1 mRNA than nonmalignant endothelial cells. S1P1 protein was present in HSA tissues and cell lines. HSA cells appeared to produce low levels of S1P, but they selectively consumed S1P from the culture media. Exogenous S1P induced an increase in intracellular calcium as well as increased proliferation and viability of HSA cells. Prolonged treatment with FTY720, an inhibitor of S1P1 , decreased S1P1 protein expression and induced apoptosis of HSA cells. S1P/S1P1 signaling pathway functions to maintain HSA cell viability and proliferation. The data suggest that S1P1 or the S1P pathway in general could be targets for therapeutic intervention for dogs with HSA. Copyright © 2015 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  5. Sugar Price Supports and Taxation

    Science.gov (United States)

    Dilk, Abby; Savaiano, Dennis A.

    2017-01-01

    Domestic US sugar production has been protected by government policy for the past 82 years, resulting in elevated domestic prices and an estimated annual (2013) $1.4 billion dollar “tax” on consumers. These elevated prices and the simultaneous federal support for domestic corn production have ensured a strong market for high-fructose corn syrup. Americans have dramatically increased their consumption of caloric sweeteners during the same period. Consumption of “empty” calories (ie, foods with low-nutrient/high-caloric density)—sugar and high-fructose corn syrup being the primary sources—is considered by most public health experts to be a key contributing factor to the rise in obesity. There have been substantial efforts to tax sugar-sweetened beverages (SSBs) to both reduce consumption and provide a source of funds for nutrition education, thereby emulating the tobacco tax model. Volume-based SSB taxes levy the tax rate per ounce of liquid, where some are only imposed on beverages with added sugar content exceeding a set threshold. Nonetheless, volume-based taxes have significant limitations in encouraging consumers to reduce their caloric intake due to a lack of transparency at the point of purchase. Thus, it is hypothesized that point-of-purchase, nutrient-specific excise taxes on SSBs would be more effective at reducing sugar consumption. However, all SSB taxes are limited by the possibility that consumers may compensate their decreased intake from SSBs with other high-calorie junk foods. Furthermore, there are no existing studies to provide evidence on how SSB taxes will impact obesity rates in the long term. The paradox of sugar prices is that Americans have paid higher prices for sugar to protect domestic production for more than 80 years, and now, Americans are being asked to pay even more to promote public health. The effective use of sugar taxes should be considered based on their merits in reducing sugar consumption and making available a new

  6. Novel Platinum-Catalyzed Ring-Opening of 1,2-Cyclopropanated Sugars with Alcohols

    DEFF Research Database (Denmark)

    Beyer, Jürgen; Madsen, Robert

    1998-01-01

    Reaction of 1,2-cyclopropanated sugars with a catalytic amount ofZeise's dimer [Pt(C2H4)Cl2]2 and an alcohol gives 2-C-branched glycosides by a novel platinum catalyzed ring-opening. A wide variety of alcohols can participate in this ring-opening reaction giving 2-C-branched glycosides ranging from...

  7. Inorganic elements in sugar samples

    International Nuclear Information System (INIS)

    Salles, Paulo M.B. de; Campos, Tarcisio P.R. de

    2013-01-01

    Sugar is considered a safe food ingredient; however, it can be contaminated by organic elements since its planting until its production process. Thus, this study aims at checking the presence of inorganic elements in samples of crystal, refined and brown sugar available for consumption in Brazil. The applied technique was neutron activation analysis, the k 0 method, using the TRIGA MARK - IPR-R1 reactor located at CDTN/CNEN, in Belo Horizonte. It was identified the presence of elements such as, Au, Br, Co, Cr, Hf, K, Na, Sb, Sc and Zn in the samples of crystal/refined sugar and the presence of As, Au, Br, Ca, Co, Cr, Cs, Fe, Hf, K, Na, Sb, Sc, Sm, Sr, Th and Zn in the brown sugar samples. The applied technique was appropriate to this study because it was not necessary to put the samples in solution, essential condition in order to apply other techniques, avoiding contaminations and sample losses, besides allowing a multi elementary detection in different sugar samples. (author)

  8. Inorganic elements in sugar samples

    Energy Technology Data Exchange (ETDEWEB)

    Salles, Paulo M.B. de; Campos, Tarcisio P.R. de, E-mail: pauladesalles@yahoo.com.br, E-mail: tprcampos@pq.cnpq.br [Universidade Federal de Minas Gerais (DEN/UFMG), Belo Horizonte, MG (Brazil). Departamento de Engenharia Nuclear; Menezes, Maria Angela de B.C., E-mail: menezes@cdtn.br [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)

    2013-07-01

    Sugar is considered a safe food ingredient; however, it can be contaminated by organic elements since its planting until its production process. Thus, this study aims at checking the presence of inorganic elements in samples of crystal, refined and brown sugar available for consumption in Brazil. The applied technique was neutron activation analysis, the k{sub 0} method, using the TRIGA MARK - IPR-R1 reactor located at CDTN/CNEN, in Belo Horizonte. It was identified the presence of elements such as, Au, Br, Co, Cr, Hf, K, Na, Sb, Sc and Zn in the samples of crystal/refined sugar and the presence of As, Au, Br, Ca, Co, Cr, Cs, Fe, Hf, K, Na, Sb, Sc, Sm, Sr, Th and Zn in the brown sugar samples. The applied technique was appropriate to this study because it was not necessary to put the samples in solution, essential condition in order to apply other techniques, avoiding contaminations and sample losses, besides allowing a multi elementary detection in different sugar samples. (author)

  9. Feedback Inhibition of Starch Degradation in Arabidopsis Leaves Mediated by Trehalose 6-Phosphate1[W][OPEN

    Science.gov (United States)

    Martins, Marina Camara Mattos; Hejazi, Mahdi; Fettke, Joerg; Steup, Martin; Feil, Regina; Krause, Ursula; Arrivault, Stéphanie; Vosloh, Daniel; Figueroa, Carlos María; Ivakov, Alexander; Yadav, Umesh Prasad; Piques, Maria; Metzner, Daniela; Stitt, Mark; Lunn, John Edward

    2013-01-01

    Many plants accumulate substantial starch reserves in their leaves during the day and remobilize them at night to provide carbon and energy for maintenance and growth. In this paper, we explore the role of a sugar-signaling metabolite, trehalose-6-phosphate (Tre6P), in regulating the accumulation and turnover of transitory starch in Arabidopsis (Arabidopsis thaliana) leaves. Ethanol-induced overexpression of trehalose-phosphate synthase during the day increased Tre6P levels up to 11-fold. There was a transient increase in the rate of starch accumulation in the middle of the day, but this was not linked to reductive activation of ADP-glucose pyrophosphorylase. A 2- to 3-fold increase in Tre6P during the night led to significant inhibition of starch degradation. Maltose and maltotriose did not accumulate, suggesting that Tre6P affects an early step in the pathway of starch degradation in the chloroplasts. Starch granules isolated from induced plants had a higher orthophosphate content than granules from noninduced control plants, consistent either with disruption of the phosphorylation-dephosphorylation cycle that is essential for efficient starch breakdown or with inhibition of starch hydrolysis by β-amylase. Nonaqueous fractionation of leaves showed that Tre6P is predominantly located in the cytosol, with estimated in vivo Tre6P concentrations of 4 to 7 µm in the cytosol, 0.2 to 0.5 µm in the chloroplasts, and 0.05 µm in the vacuole. It is proposed that Tre6P is a component in a signaling pathway that mediates the feedback regulation of starch breakdown by sucrose, potentially linking starch turnover to demand for sucrose by growing sink organs at night. PMID:24043444

  10. European Multicenter Study on Analytical Performance of Veris HIV-1 Assay.

    Science.gov (United States)

    Braun, Patrick; Delgado, Rafael; Drago, Monica; Fanti, Diana; Fleury, Hervé; Hofmann, Jörg; Izopet, Jacques; Kalus, Ulrich; Lombardi, Alessandra; Marcos, Maria Angeles; Mileto, Davide; Sauné, Karine; O'Shea, Siobhan; Pérez-Rivilla, Alfredo; Ramble, John; Trimoulet, Pascale; Vila, Jordi; Whittaker, Duncan; Artus, Alain; Rhodes, Daniel W

    2017-07-01

    The analytical performance of the Veris HIV-1 assay for use on the new, fully automated Beckman Coulter DxN Veris molecular diagnostics system was evaluated at 10 European virology laboratories. The precision, analytical sensitivity, performance with negative samples, linearity, and performance with HIV-1 groups/subtypes were evaluated. The precision for the 1-ml assay showed a standard deviation (SD) of 0.14 log 10 copies/ml or less and a coefficient of variation (CV) of ≤6.1% for each level tested. The 0.175-ml assay showed an SD of 0.17 log 10 copies/ml or less and a CV of ≤5.2% for each level tested. The analytical sensitivities determined by probit analysis were 19.3 copies/ml for the 1-ml assay and 126 copies/ml for the 0.175-ml assay. The performance with 1,357 negative samples demonstrated 99.2% with not detected results. Linearity using patient samples was shown from 1.54 to 6.93 log 10 copies/ml. The assay performed well, detecting and showing linearity with all HIV-1 genotypes tested. The Veris HIV-1 assay demonstrated analytical performance comparable to that of currently marketed HIV-1 assays. (DxN Veris products are Conformité Européenne [CE]-marked in vitro diagnostic products. The DxN Veris product line has not been submitted to the U.S. FDA and is not available in the U.S. market. The DxN Veris molecular diagnostics system is also known as the Veris MDx molecular diagnostics system and the Veris MDx system.). Copyright © 2017 American Society for Microbiology.

  11. Serum-Sphingosine-1-Phosphate Concentrations Are Inversely Associated with Atherosclerotic Diseases in Humans.

    Directory of Open Access Journals (Sweden)

    Irina Soltau

    Full Text Available Atherosclerotic changes of arteries are the leading cause for deaths in cardiovascular disease and greatly impair patient's quality of life. Sphingosine-1-phosphate (S1P is a signaling sphingolipid that regulates potentially pro-as well as anti-atherogenic processes. Here, we investigate whether serum-S1P concentrations are associated with peripheral artery disease (PAD and carotid stenosis (CS.Serum was sampled from blood donors (controls, N = 174 and from atherosclerotic patients (N = 132 who presented to the hospital with either clinically relevant PAD (N = 102 or CS (N = 30. From all subjects, serum-S1P was measured by mass spectrometry and blood parameters were determined by routine laboratory assays. When compared to controls, atherosclerotic patients before invasive treatment to restore blood flow showed significantly lower serum-S1P levels. This difference cannot be explained by risk factors for atherosclerosis (old age, male gender, hypertension, hypercholesteremia, obesity, diabetes or smoking or comorbidities (Chronic obstructive pulmonary disease, kidney insufficiency or arrhythmia. Receiver operating characteristic curves suggest that S1P has more power to indicate atherosclerosis (PAD and CS than high density lipoprotein-cholesterol (HDL-C. In 35 patients, serum-S1P was measured again between one and six months after treatment. In this group, serum-S1P concentrations rose after treatment independent of whether patients had PAD or CS, or whether they underwent open or endovascular surgery. Post-treatment S1P levels were highly associated to platelet numbers measured pre-treatment.Our study shows that PAD and CS in humans is associated with decreased serum-S1P concentrations and that S1P may possess higher accuracy to indicate these diseases than HDL-C.

  12. Use of Added Sugars Instead of Total Sugars May Improve the Capacity of the Health Star Rating System to Discriminate between Core and Discretionary Foods.

    Science.gov (United States)

    Menday, Hannah; Neal, Bruce; Wu, Jason H Y; Crino, Michelle; Baines, Surinder; Petersen, Kristina S

    2017-12-01

    The Australian Government has introduced a voluntary front-of-package labeling system that includes total sugar in the calculation. Our aim was to determine the effect of substituting added sugars for total sugars when calculating Health Star Ratings (HSR) and identify whether use of added sugars improves the capacity to distinguish between core and discretionary food products. This study included packaged food and beverage products available in Australian supermarkets (n=3,610). The product categories included in the analyses were breakfast cereals (n=513), fruit (n=571), milk (n=309), non-alcoholic beverages (n=1,040), vegetables (n=787), and yogurt (n=390). Added sugar values were estimated for each product using a validated method. HSRs were then estimated for every product according to the established method using total sugar, and then by substituting added sugar for total sugar. The scoring system was not modified when added sugar was used in place of total sugar in the HSR calculation. Products were classified as core or discretionary based on the Australian Dietary Guidelines. To investigate whether use of added sugar in the HSR algorithm improved the distinction between core and discretionary products as defined by the Australian Dietary Guidelines, the proportion of core products that received an HSR of ≥3.5 stars and the proportion of discretionary products that received an HSR of added sugars were determined. There were 2,263 core and 1,347 discretionary foods; 1,684 of 3,610 (47%) products contained added sugar (median 8.4 g/100 g, interquartile range=5.0 to 12.2 g). When the HSR was calculated with added sugar instead of total sugar, an additional 166 (7.3%) core products received an HSR of ≥3.5 stars and 103 (7.6%) discretionary products received a rating of ≥3.5 stars. The odds of correctly identifying a product as core vs discretionary were increased by 61% (odds ratio 1.61, 95% CI 1.26 to 2.06; Padded compared to total sugars. In the six

  13. Reduction of Saltiness and Acrylamide Levels in Palm Sugar-Like Flavouring through Buffer Modification and the Addition of Calcium Chloride

    Directory of Open Access Journals (Sweden)

    Wan Aida Wan Mustapha

    2013-06-01

    Full Text Available Palm sugar-like flavouring (PSLF is a type of flavour product that is formed by heating amino acids and sugar under specific heating conditions. Unfortunately, PSLF has a salty taste and contains high amounts of acrylamide. Hence, the objective of this research was to reduce saltiness and acrylamide without negatively affecting the aroma properties of PSLF. A decrease in the sodium phosphate (NaHPO4 buffer concentration from 0.20 to 0.02 M was found to reduce sodium to approximately 15% of the level found in original PSLF. A further decrease (~25% in the sodium content was achieved by removing monobasic sodium phosphate (NaH2PO4 from the buffer system. Meanwhile, the addition of CaCl2 at 20–40 mg/L reduced the acrylamide content in PSLF by as much as 58%. A CaCl2 concentration of 20 mg/mL was most favourable as it most efficiently suppressed acrylamide formation while providing an acceptably high flavour yield in PSLF. In view of the high acrylamide content in PSLF, additional work is necessary to further reduce the amount of acrylamide by controlling the asparagine concentration in the precursor mixture.

  14. Reduction of saltiness and acrylamide levels in palm sugar-like flavouring through buffer modification and the addition of calcium chloride.

    Science.gov (United States)

    Tan, Phui Yee; Tan, Chin Ping; Abas, Faridah; Ho, Chun Wai; Mustapha, Wan Aida Wan

    2013-06-10

    Palm sugar-like flavouring (PSLF) is a type of flavour product that is formed by heating amino acids and sugar under specific heating conditions. Unfortunately, PSLF has a salty taste and contains high amounts of acrylamide. Hence, the objective of this research was to reduce saltiness and acrylamide without negatively affecting the aroma properties of PSLF. A decrease in the sodium phosphate (NaHPO₄) buffer concentration from 0.20 to 0.02 M was found to reduce sodium to approximately 15% of the level found in original PSLF. A further decrease (~25%) in the sodium content was achieved by removing monobasic sodium phosphate (NaH₂PO₄) from the buffer system. Meanwhile, the addition of CaCl₂ at 20-40 mg/L reduced the acrylamide content in PSLF by as much as 58%. A CaCl₂ concentration of 20 mg/mL was most favourable as it most efficiently suppressed acrylamide formation while providing an acceptably high flavour yield in PSLF. In view of the high acrylamide content in PSLF, additional work is necessary to further reduce the amount of acrylamide by controlling the asparagine concentration in the precursor mixture.

  15. Early detection of sugar beet pathogen Ramularia beticola in leaf and air samples using qPCR

    DEFF Research Database (Denmark)

    Wieczorek, Thies Marten; Jørgensen, Lise Nistrup; Hansen, Anne Lisbet

    2014-01-01

    A quantitative PCR method (qPCR) was developed for the detection and quantification of Ramularia beticola causing Ramularia leaf spot in sugar beet. R. beticola specific primers were designed based on the internal transcribed spacer region 2 (ITS2). The assay was applied on DNA extracted from...... spores trapped on tape from Burkard spore traps placed in an artificially inoculated sugar beet field trial and in two sugar beet fields with natural infections. R. beticola DNA was detected at variable amounts in the air samples 14 to 16 days prior to first visible symptoms. R. beticola DNA was detected...... in air samples from fields with natural infection at significant and increasing levels from development of the first symptoms, indicating that spore production within the crop plays a major role in the epidemic development of the disease. Sugar beet leaves sampled from the inoculated field trial were...

  16. A homogeneous, high-throughput assay for phosphatidylinositol 5-phosphate 4-kinase with a novel, rapid substrate preparation.

    Directory of Open Access Journals (Sweden)

    Mindy I Davis

    Full Text Available Phosphoinositide kinases regulate diverse cellular functions and are important targets for therapeutic development for diseases, such as diabetes and cancer. Preparation of the lipid substrate is crucial for the development of a robust and miniaturizable lipid kinase assay. Enzymatic assays for phosphoinositide kinases often use lipid substrates prepared from lyophilized lipid preparations by sonication, which result in variability in the liposome size from preparation to preparation. Herein, we report a homogeneous 1536-well luciferase-coupled bioluminescence assay for PI5P4Kα. The substrate preparation is novel and allows the rapid production of a DMSO-containing substrate solution without the need for lengthy liposome preparation protocols, thus enabling the scale-up of this traditionally difficult type of assay. The Z'-factor value was greater than 0.7 for the PI5P4Kα assay, indicating its suitability for high-throughput screening applications. Tyrphostin AG-82 had been identified as an inhibitor of PI5P4Kα by assessing the degree of phospho transfer of γ-(32P-ATP to PI5P; its inhibitory activity against PI5P4Kα was confirmed in the present miniaturized assay. From a pilot screen of a library of bioactive compounds, another tyrphostin, I-OMe tyrphostin AG-538 (I-OMe-AG-538, was identified as an ATP-competitive inhibitor of PI5P4Kα with an IC(50 of 1 µM, affirming the suitability of the assay for inhibitor discovery campaigns. This homogeneous assay may apply to other lipid kinases and should help in the identification of leads for this class of enzymes by enabling high-throughput screening efforts.

  17. DEVELOPMENT OF A METHOD FOR QUANTITATING SPHINGOID BASE 1-PHOSPHATES IN BLOOD SPOTS

    Science.gov (United States)

    Red blood cells (RBC) accumulate, store and release sphingoid base 1-phosphates,important ligands for the extracellular receptors S1P1-5. The ability of RBC to accumulate these bioactive lipids is because, with the exception of sphingosine kinase, the enzymes responsible for metabolizing sphingosine...

  18. Quantitative monitoring of sucrose, reducing sugar and total sugar dynamics for phenotyping of water-deficit stress tolerance in rice through spectroscopy and chemometrics

    Science.gov (United States)

    Das, Bappa; Sahoo, Rabi N.; Pargal, Sourabh; Krishna, Gopal; Verma, Rakesh; Chinnusamy, Viswanathan; Sehgal, Vinay K.; Gupta, Vinod K.; Dash, Sushanta K.; Swain, Padmini

    2018-03-01

    In the present investigation, the changes in sucrose, reducing and total sugar content due to water-deficit stress in rice leaves were modeled using visible, near infrared (VNIR) and shortwave infrared (SWIR) spectroscopy. The objectives of the study were to identify the best vegetation indices and suitable multivariate technique based on precise analysis of hyperspectral data (350 to 2500 nm) and sucrose, reducing sugar and total sugar content measured at different stress levels from 16 different rice genotypes. Spectral data analysis was done to identify suitable spectral indices and models for sucrose estimation. Novel spectral indices in near infrared (NIR) range viz. ratio spectral index (RSI) and normalised difference spectral indices (NDSI) sensitive to sucrose, reducing sugar and total sugar content were identified which were subsequently calibrated and validated. The RSI and NDSI models had R2 values of 0.65, 0.71 and 0.67; RPD values of 1.68, 1.95 and 1.66 for sucrose, reducing sugar and total sugar, respectively for validation dataset. Different multivariate spectral models such as artificial neural network (ANN), multivariate adaptive regression splines (MARS), multiple linear regression (MLR), partial least square regression (PLSR), random forest regression (RFR) and support vector machine regression (SVMR) were also evaluated. The best performing multivariate models for sucrose, reducing sugars and total sugars were found to be, MARS, ANN and MARS, respectively with respect to RPD values of 2.08, 2.44, and 1.93. Results indicated that VNIR and SWIR spectroscopy combined with multivariate calibration can be used as a reliable alternative to conventional methods for measurement of sucrose, reducing sugars and total sugars of rice under water-deficit stress as this technique is fast, economic, and noninvasive.

  19. Biomimetic synthesis of modified calcium phosphate fine powders and their in vitro studies

    Energy Technology Data Exchange (ETDEWEB)

    Gergulova, R., E-mail: rumigg@yahoo.com; Tepavitcharova, S., E-mail: rumigg@yahoo.com; Rabadjieva, D., E-mail: rumigg@yahoo.com; Sezanova, K., E-mail: rumigg@yahoo.com; Ilieva, R., E-mail: rumigg@yahoo.com [Institute of General and Inorganic Chemistry, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 11, 1113 Sofia (Bulgaria); Alexandrova, R.; Andonova-Lilova, B. [Institute of Experimental Morphology, Pathology and Anthropology with Museum, BAS, Acad. G. Bonchev Str., Bl. 25, Sofia (Bulgaria)

    2013-12-16

    Biomimetic approach and subsequent high-temperature treatment were used to synthesize ion modified calcium phosphate fine powders. Thus, using Simulated Body Fluid (SBF) as an ion modifier, a bi-phase mixture of ion modified β-tricalcium phosphate and hydroxyapatite (β-TCP + HA) was prepared. The use of SBF electrolyte solution enriched with Mg{sup 2+} or Zn{sup 2+} yielded monophase β-tricalcium phosphate additionally modified with Mg{sup 2+} or Zn{sup 2+} (Mg-β-TCP or Zn-β-TCP). The in vitro behavior of the prepared powders on cell viability and proliferation of murine BALB/c 3T3 fibroblasts and of human Lep 3 cells was studied by MTT test assays and Mosmann method after 72 h incubation. The relative cell viability was calculated.

  20. Urinary sugars biomarker relates better to extrinsic than to intrinsic sugars intake in a metabolic study with volunteers consuming their normal diet.

    Science.gov (United States)

    Tasevska, N; Runswick, S A; Welch, A A; McTaggart, A; Bingham, S A

    2009-05-01

    Sugars in diet are very difficult to measure because of the unreliability of self-reported dietary intake. Sucrose and fructose excreted in urine have been recently suggested as a biomarker for total sugars intake. To further characterize the use of this biomarker, we investigated whether urinary sugars correlated better to extrinsic compared to intrinsic sugars in the diet. Seven male and six female healthy participants were living for 30 days in a metabolic suite under strictly controlled conditions consuming their usual diet as assessed beforehand from four consecutive 7-day food diaries kept at home. During the 30-day study, all 24 h urine specimens were collected, validated for their completeness and analysed for sucrose and fructose. The mean total sugars intake in the group was 202+/-69 g day(-1). Daily intake of extrinsic, intrinsic and milk sugars contributed 60.1, 34.4 and 5.5%, to the total sugars intake, respectively. The individuals' 30-day mean sugars excretion levels were significantly correlated with the 30-day means of extrinsic sugars (r=0.84; Psugars intake (r=0.43; P=0.144). In the regression, only extrinsic sugars intake explained a significant proportion of the variability in sugars excretion (adjusted R(2)=0.64; P=0.001); daily excretion of 100 mg sucrose and fructose in urine predicted 124 g of extrinsic total sugars in the diet. Using fewer urinary and dietary measurements in the analysis did not change the overall trend of the findings. In this group of volunteers, sucrose and fructose in urine better correlated to extrinsic than to intrinsic sugars intake.

  1. Application of chitosan/polyacrylamide nanofibres for removal of chromate and phosphate in water

    Science.gov (United States)

    Nthumbi, Richard M.; Catherine Ngila, J.; Moodley, Brenda; Kindness, Andrew; Petrik, Leslie

    Water pollution is an intractable environmental problem in South Africa. Management of the water resource is vital in order to address the water scarcity issues. Research on remediation of contaminated water has focused mainly on the removal of heavy metals such as Pb, Cd, Zn, Hg and Cu and neglected other inorganic pollutants. In this work we focus on the removal of anions, namely chromate and phosphate. Chromium is extensively used in the textile, leather and metallurgy industries and contaminates surface water and groundwater when inadequately treated industrial effluents are discharged. Chromium has been associated with irregular sugar metabolism, nosebleeds and ulcers, and it is also carcinogenic. The phosphate ion is an essential micronutrient responsible for healthy plant growth. However, excess phosphate intake stimulates rapid growth of photosynthetic algae and cyanobacteria, resulting in eutrophication. This phenomenon (algal bloom) causes other organisms to die due to reduced oxygen in the water. In order to offer remediation measures, this study reports the use of electrospun nanofibres for the removal of chromate and phosphate anions. Adsorption experiments were carried out using nanofibres electrospun from chitosan and polyacrylamide polymer blends, cross-linked with glutaraldehyde. Quantification of chromium was done using ICP-OES while UV-Vis spectrophotometry was used for the determination of phosphates. Batch adsorption experiments were done to determine optimum adsorption parameters such as pH, contact time, temperature and initial analyte concentration. Removal of the ions using a flow-adsorption technique through a micro-column was performed. The experimental data obtained were analysed using Langmuir and Freundlich models to study the adsorption mechanisms. The nanofibres had an adsorption capacity for Cr(VI) and PO43- of 0.26 mg g-1 and 392 mg g-1, respectively, and removal efficiencies of 93% and 97.4%, in the same order, in synthetic water

  2. The Truth about Sugar.

    Science.gov (United States)

    Yeung, C Albert; Goodfellow, Ashley; Flanagan, Louise

    2015-01-01

    Sugars are used by the industry to enhance the attractiveness of foods and drinks. These added sugars, or 'free sugars', are not easily identified in food or drink labels. Certain manufactured foods and drinks with 'safe' names, such as dried fruit and fruit juice, still contain free sugars and can be confusing. Guidance states that daily consumption of free sugars should be less than 10% of total energy intake (no more than 5% in the UK). However, it is found that both tooth decay and obesity are associated with consumption of free sugars in large quantities and at inappropriate times.

  3. Comparative transcriptome analysis reveals key genes potentially related to soluble sugar and organic acid accumulation in watermelon

    Science.gov (United States)

    Gao, Lei; Zhao, Shengjie; Lu, Xuqiang; He, Nan; Zhu, Hongju; Dou, Junling

    2018-01-01

    Soluble sugars and organic acids are important components of fruit flavor and have a strong impact on the overall organoleptic quality of watermelon (Citrullus lanatus) fruit. Several studies have analyzed the expression levels of the genes related to soluble sugar accumulation and the dynamic changes in their content during watermelon fruit development and ripening. Nevertheless, to date, there have been no reports on the organic acid content in watermelon or the genes regulating their synthesis. In this study, the soluble sugars and organic acids in watermelon were measured and a comparative transcriptome analysis was performed to identify the key genes involved in the accumulation of these substances during fruit development and ripening. The watermelon cultivar ‘203Z’ and its near-isogenic line (NIL) ‘SW’ (in the ‘203Z’ background) were used as experimental materials. The results suggested that soluble sugar consist of fructose, glucose and sucrose while malic-, citric-, and oxalic acids are the primary organic acids in watermelon fruit. Several differentially expressed genes (DEGs) related to soluble sugar- and organic acid accumulation and metabolism were identified. These include the DEGs encoding raffinose synthase, sucrose synthase (SuSy), sucrose-phosphate synthase (SPSs), insoluble acid invertases (IAI), NAD-dependent malate dehydrogenase (NAD-cyt MDH), aluminum-activated malate transporter (ALMT), and citrate synthase (CS). This is the first report addressing comparative transcriptome analysis via NILs materials in watermelon fruit. These findings provide an important basis for understanding the molecular mechanism that leads to soluble sugar and organic acid accumulation and metabolism during watermelon fruit development and ripening. PMID:29324867

  4. Comparative transcriptome analysis reveals key genes potentially related to soluble sugar and organic acid accumulation in watermelon.

    Directory of Open Access Journals (Sweden)

    Lei Gao

    Full Text Available Soluble sugars and organic acids are important components of fruit flavor and have a strong impact on the overall organoleptic quality of watermelon (Citrullus lanatus fruit. Several studies have analyzed the expression levels of the genes related to soluble sugar accumulation and the dynamic changes in their content during watermelon fruit development and ripening. Nevertheless, to date, there have been no reports on the organic acid content in watermelon or the genes regulating their synthesis. In this study, the soluble sugars and organic acids in watermelon were measured and a comparative transcriptome analysis was performed to identify the key genes involved in the accumulation of these substances during fruit development and ripening. The watermelon cultivar '203Z' and its near-isogenic line (NIL 'SW' (in the '203Z' background were used as experimental materials. The results suggested that soluble sugar consist of fructose, glucose and sucrose while malic-, citric-, and oxalic acids are the primary organic acids in watermelon fruit. Several differentially expressed genes (DEGs related to soluble sugar- and organic acid accumulation and metabolism were identified. These include the DEGs encoding raffinose synthase, sucrose synthase (SuSy, sucrose-phosphate synthase (SPSs, insoluble acid invertases (IAI, NAD-dependent malate dehydrogenase (NAD-cyt MDH, aluminum-activated malate transporter (ALMT, and citrate synthase (CS. This is the first report addressing comparative transcriptome analysis via NILs materials in watermelon fruit. These findings provide an important basis for understanding the molecular mechanism that leads to soluble sugar and organic acid accumulation and metabolism during watermelon fruit development and ripening.

  5. Comparative transcriptome analysis reveals key genes potentially related to soluble sugar and organic acid accumulation in watermelon.

    Science.gov (United States)

    Gao, Lei; Zhao, Shengjie; Lu, Xuqiang; He, Nan; Zhu, Hongju; Dou, Junling; Liu, Wenge

    2018-01-01

    Soluble sugars and organic acids are important components of fruit flavor and have a strong impact on the overall organoleptic quality of watermelon (Citrullus lanatus) fruit. Several studies have analyzed the expression levels of the genes related to soluble sugar accumulation and the dynamic changes in their content during watermelon fruit development and ripening. Nevertheless, to date, there have been no reports on the organic acid content in watermelon or the genes regulating their synthesis. In this study, the soluble sugars and organic acids in watermelon were measured and a comparative transcriptome analysis was performed to identify the key genes involved in the accumulation of these substances during fruit development and ripening. The watermelon cultivar '203Z' and its near-isogenic line (NIL) 'SW' (in the '203Z' background) were used as experimental materials. The results suggested that soluble sugar consist of fructose, glucose and sucrose while malic-, citric-, and oxalic acids are the primary organic acids in watermelon fruit. Several differentially expressed genes (DEGs) related to soluble sugar- and organic acid accumulation and metabolism were identified. These include the DEGs encoding raffinose synthase, sucrose synthase (SuSy), sucrose-phosphate synthase (SPSs), insoluble acid invertases (IAI), NAD-dependent malate dehydrogenase (NAD-cyt MDH), aluminum-activated malate transporter (ALMT), and citrate synthase (CS). This is the first report addressing comparative transcriptome analysis via NILs materials in watermelon fruit. These findings provide an important basis for understanding the molecular mechanism that leads to soluble sugar and organic acid accumulation and metabolism during watermelon fruit development and ripening.

  6. Sugar-Sweetened Beverages Are the Main Sources of Added Sugar Intake in the Mexican Population.

    Science.gov (United States)

    Sánchez-Pimienta, Tania G; Batis, Carolina; Lutter, Chessa K; Rivera, Juan A

    2016-09-01

    Sugar intake has been associated with an increased prevalence of obesity, other noncommunicable diseases, and dental caries. The WHO recommends that free sugars should be ENSANUT (National Health and Nutrition Survey) 2012], which represents 3 geographic regions and urban and rural areas. Dietary information was obtained by administering a 24-h recall questionnaire to 10,096 participants. Total sugar intake was estimated by using the National Institute of Public Health (INSP) food-composition table and an established method to estimate added sugars. The mean intakes of total, intrinsic, and added sugars were 365, 127, and 238 kcal/d, respectively. Added sugars contributed 13% of TEI. Sugar-sweetened beverages (SSBs) were the main source of sugars, contributing 69% of added sugars. Food products high in saturated fat and/or added sugar (HSFAS) were the second main sources of added sugars, contributing 25% of added sugars. The average intake of added sugars in the Mexican diet is higher than WHO recommendations, which may partly explain the high prevalence of obesity and diabetes in Mexico. Because SSBs and HSFAS contribute >94% of total added sugars, strategies to reduce their intake should be strengthened. This includes stronger food labels to warn the consumer about the content of added sugars in foods and beverages. © 2016 American Society for Nutrition.

  7. Some observations on phosphate based corrosion inhibitors in preventing carbon steel corrosion

    International Nuclear Information System (INIS)

    Anupkumar, B.; Satpathy, K.K.

    2000-01-01

    Among the various types of phosphonic acid based inhibitors assayed, namely HEDP, ATMP and a commercial corrosion inhibitor (code named Betz), it was found that Betz has the maximum amount of organic phosphate followed by HEDP and ATMP. The corrosion rate studies show that Betz gives the highest inhibition efficiency followed by HEDP and ATMP. This shows that organic phosphate plays a significant role in corrosion protection. However, it was observed that due to synergestic effect, HEDP in the presence of Zn 2+ gave a better corrosion protection than Betz. The results are discussed in the light of available literature. (author)

  8. Effects of serving high-sugar cereals on children's breakfast-eating behavior.

    Science.gov (United States)

    Harris, Jennifer L; Schwartz, Marlene B; Ustjanauskas, Amy; Ohri-Vachaspati, Punam; Brownell, Kelly D

    2011-01-01

    To test (1) whether children will consume low-sugar ready-to-eat (RTE) cereals and (2) the effects of serving high- versus low-sugar cereals on the consumption of cereal, refined sugar, fresh fruit, and milk. Using an experimental design, we randomly assigned children (n = 91) who were attending summer day camp to receive a breakfast that included either the choice of 1 of 3 high-sugar cereals (high-sugar condition) or low-sugar cereals (low-sugar condition), as well as low-fat milk, orange juice, bananas, strawberries, and sugar packets. Participants served themselves and completed a background questionnaire after eating. Researchers measured the amount and calories consumed of each food. In both conditions, children reported "liking" or "loving" the cereal they chose. Children in the low-sugar cereal condition consumed, on average, slightly more than 1 serving of cereal (35 g), whereas children in the high-sugar condition consumed significantly more (61 g) and almost twice the amount of refined sugar in total (24.4 vs 12.5 g). Milk and total calories consumed did not differ significantly between conditions, but children in the low-sugar condition were more likely to put fruit on their cereal (54% vs 8%) and consumed a greater portion of total calories from fresh fruit (20% vs 13%). Compared with serving low-sugar cereals, high-sugar cereals increase children's total sugar consumption and reduce the overall nutritional quality of their breakfast. Children will consume low-sugar cereals when offered, and they provide a superior breakfast option.

  9. Motile hepatocellular carcinoma cells preferentially secret sugar metabolism regulatory proteins via exosomes.

    Science.gov (United States)

    Zhang, Jing; Lu, Shaohua; Zhou, Ye; Meng, Kun; Chen, Zhipeng; Cui, Yizhi; Shi, Yunfeng; Wang, Tong; He, Qing-Yu

    2017-07-01

    Exosomes are deliverers of critically functional proteins, capable of transforming target cells in numerous cancers, including hepatocellular carcinoma (HCC). We hypothesize that the motility of HCC cells can be featured by comparative proteome of exosomes. Hence, we performed the super-SILAC-based MS analysis on the exosomes secreted by three human HCC cell lines, including the non-motile Hep3B cell, and the motile 97H and LM3 cells. More than 1400 exosomal proteins were confidently quantified in each MS analysis with highly biological reproducibility. We justified that 469 and 443 exosomal proteins represented differentially expressed proteins (DEPs) in the 97H/Hep3B and LM3/Hep3B comparisons, respectively. These DEPs focused on sugar metabolism-centric canonical pathways per ingenuity pathway analysis, which was consistent with the gene ontology analysis on biological process enrichment. These pathways included glycolysis I, gluconeogenesis I and pentose phosphate pathways; and the DEPs enriched in these pathways could form a tightly connected network. By analyzing the relative abundance of proteins and translating mRNAs, we found significantly positive correlation between exosomes and cells. The involved exosomal proteins were again focusing on sugar metabolism. In conclusion, motile HCC cells tend to preferentially export more sugar metabolism-associated proteins via exosomes that differentiate them from non-motile HCC cells. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Association of Dietary Sugars and Sugar-Sweetened Beverage Intake with Obesity in Korean Children and Adolescents

    Directory of Open Access Journals (Sweden)

    Kyungho Ha

    2016-01-01

    Full Text Available Few studies have examined the association between dietary sugar intake and obesity in Asian children and adolescents. We evaluated the association of dietary sugar intake and its food source with obesity in Korean children and adolescents. In this cross-sectional analysis, data were obtained from five studies conducted between 2002 and 2011. The study included 2599 children and adolescents who had completed more than three days of dietary records and had anthropometric data. Total sugar intake was higher in girls than in boys (54.3 g for girls and 46.6 g for boys, p < 0.0001. Sugar intake from milk and fruits was inversely associated with overweight or obesity in girls only (OR for overweight, 0.52; 95% CI, 0.32–0.84; p for trend = 0.0246 and OR for obesity, 0.42; 95% CI, 0.23–0.79; p for trend = 0.0113. Sugar-sweetened beverage (SSB consumption was not associated with obesity in girls, while boys had lower odds ratios for obesity (OR for obesity, 0.52; 95% CI, 0.26–1.05; p for trend = 0.0310. These results suggest that total sugars and SSB intake in Asian children and adolescents remains relatively low and sugar intake from milk and fruits is associated with a decreased risk of overweight or obesity, especially in girls.

  11. Association of Dietary Sugars and Sugar-Sweetened Beverage Intake with Obesity in Korean Children and Adolescents.

    Science.gov (United States)

    Ha, Kyungho; Chung, Sangwon; Lee, Haeng-Shin; Kim, Cho-il; Joung, Hyojee; Paik, Hee-Young; Song, YoonJu

    2016-01-08

    Few studies have examined the association between dietary sugar intake and obesity in Asian children and adolescents. We evaluated the association of dietary sugar intake and its food source with obesity in Korean children and adolescents. In this cross-sectional analysis, data were obtained from five studies conducted between 2002 and 2011. The study included 2599 children and adolescents who had completed more than three days of dietary records and had anthropometric data. Total sugar intake was higher in girls than in boys (54.3 g for girls and 46.6 g for boys, p < 0.0001). Sugar intake from milk and fruits was inversely associated with overweight or obesity in girls only (OR for overweight, 0.52; 95% CI, 0.32-0.84; p for trend = 0.0246 and OR for obesity, 0.42; 95% CI, 0.23-0.79; p for trend = 0.0113). Sugar-sweetened beverage (SSB) consumption was not associated with obesity in girls, while boys had lower odds ratios for obesity (OR for obesity, 0.52; 95% CI, 0.26-1.05; p for trend = 0.0310). These results suggest that total sugars and SSB intake in Asian children and adolescents remains relatively low and sugar intake from milk and fruits is associated with a decreased risk of overweight or obesity, especially in girls.

  12. 75 FR 60715 - Domestic Sugar Program-FY 2010 and FY 2011 Cane Sugar and Beet Sugar Marketing Allotments and...

    Science.gov (United States)

    2010-10-01

    ... marketing allotment and the associated production history will be transferred from MDFC to WSG, effective... Cane Sugar and Beet Sugar Marketing Allotments and Company Allocations AGENCY: Commodity Credit... publish the modifications to the fiscal year 2010 (FY 2010) State sugar marketing allotments and company...

  13. Different response patterns of several ligands at the sphingosine-1-phosphate receptor subtype 3 (S1P(3))

    NARCIS (Netherlands)

    Jongsma, M.; van Unen, J.; van Loenen, P. B.; Michel, M. C.; Peters, S. L. M.; Alewijnse, A. E.

    2009-01-01

    Recently, some ligands targeting the sphingosine-1-phosphate receptor subtype 3 (S1P(3)) have become available. The characterization of these compounds was mainly based on one functional read-out system, although S1P(3) receptors are known to activate different signal transduction pathways.

  14. Structure-guided mutational analysis of the OB, HhH, and BRCT domains of Escherichia coli DNA ligase.

    Science.gov (United States)

    Wang, Li Kai; Nair, Pravin A; Shuman, Stewart

    2008-08-22

    NAD(+)-dependent DNA ligases (LigAs) are ubiquitous in bacteria and essential for growth. LigA enzymes have a modular structure in which a central catalytic core composed of nucleotidyltransferase and oligonucleotide-binding (OB) domains is linked via a tetracysteine zinc finger to distal helix-hairpin-helix (HhH) and BRCT (BRCA1-like C-terminal) domains. The OB and HhH domains contribute prominently to the protein clamp formed by LigA around nicked duplex DNA. Here we conducted a structure-function analysis of the OB and HhH domains of Escherichia coli LigA by alanine scanning and conservative substitutions, entailing 43 mutations at 22 amino acids. We thereby identified essential functional groups in the OB domain that engage the DNA phosphodiester backbone flanking the nick (Arg(333)); penetrate the minor grove and distort the nick (Val(383) and Ile(384)); or stabilize the OB fold (Arg(379)). The essential constituents of the HhH domain include: four glycines (Gly(455), Gly(489), Gly(521), Gly(553)), which bind the phosphate backbone across the minor groove at the outer margins of the LigA-DNA interface; Arg(487), which penetrates the minor groove at the outer margin on the 3 (R)-OH side of the nick; and Arg(446), which promotes protein clamp formation via contacts to the nucleotidyltransferase domain. We find that the BRCT domain is required in its entirety for effective nick sealing and AMP-dependent supercoil relaxation.

  15. L-Arabinose isomerase and its use for biotechnological production of rare sugars.

    Science.gov (United States)

    Xu, Zheng; Li, Sha; Feng, Xiaohai; Liang, Jinfeng; Xu, Hong

    2014-11-01

    L-Arabinose isomerase (AI), a key enzyme in the microbial pentose phosphate pathway, has been regarded as an important biological catalyst in rare sugar production. This enzyme could isomerize L-arabinose into L-ribulose, as well as D-galactose into D-tagatose. Both the two monosaccharides show excellent commercial values in food and pharmaceutical industries. With the identification of novel AI family members, some of them have exhibited remarkable potential in industrial applications. The biological production processes for D-tagatose and L-ribose (or L-ribulose) using AI have been developed and improved in recent years. Meanwhile, protein engineering techniques involving rational design has effectively enhanced the catalytic properties of various AIs. Moreover, the crystal structure of AI has been disclosed, which sheds light on the understanding of AI structure and catalytic mechanism at molecular levels. This article reports recent developments in (i) novel AI screening, (ii) AI-mediated rare sugar production processes, (iii) molecular modification of AI, and (iv) structural biology study of AI. Based on previous reports, an analysis of the future development has also been initiated.

  16. Sphingosine 1-Phosphate Receptor 1 as a Useful Target for Treatment of Multiple Sclerosis

    Directory of Open Access Journals (Sweden)

    Kunitomo Adachi

    2012-05-01

    Full Text Available Sphingosine 1-phosphate (S1P, a lysophospholipid mediator, is generated from sphingosine by sphingosine kinases and binds five known cell surface receptors. S1P receptor 1 (S1P1 plays an essential role in lymphocyte egress from secondary lymphoid organs (SLO, as evinced by the inability of lymphocytes to exit from the SLO in mice lacking lymphocytic S1P1. Fingolimod hydrochloride (FTY720 is a first-in-class, orally active, S1P receptor modulator with a structure closely related to sphingosine. FTY720 was first synthesized by chemical modification of a natural product, myriocin. FTY720 is effectively converted to an active metabolite, FTY720 phosphate (FTY720-P by sphingosine kinases. FTY720-P shows high affinity to 4 of the S1P receptors (S1P1, S1P3, S1P4, and S1P5. In particular, FTY720-P strongly induces internalization and degradation of S1P1, inhibits S1P responsiveness of lymphocytes in the SLO, and acts as a functional antagonist at lymphocytic S1P1. Consequently, FTY720 inhibits S1P1-dependent lymphocyte egress from the SLO to decrease circulation of lymphocytes including autoreactive Th17 cells and is highly effective in experimental autoimmune encephalomyelitis (EAE, an animal model of multiple sclerosis (MS. Because FTY720 shows a superior efficacy in relapsing remitting MS patients compared to intramuscular interferon-β-1a (Avonex®, S1P1 is presumed to be a useful target for the therapy of MS.

  17. 21 CFR 173.320 - Chemicals for controlling microorganisms in cane-sugar and beet-sugar mills.

    Science.gov (United States)

    2010-04-01

    ...-sugar and beet-sugar mills. 173.320 Section 173.320 Food and Drugs FOOD AND DRUG ADMINISTRATION... controlling microorganisms in cane-sugar and beet-sugar mills. Agents for controlling microorganisms in cane-sugar and beet-sugar mills may be safely used in accordance with the following conditions: (a) They are...

  18. Transcript profiles uncover temporal and stress-induced changes of metabolic pathways in germinating sugar beet seeds

    Directory of Open Access Journals (Sweden)

    Windhövel Andrea

    2008-12-01

    Full Text Available Abstract Background With a cultivation area of 1.75 Mio ha and sugar yield of 16.7 Mio tons in 2006, sugar beet is a crop of great economic importance in Europe. The productivity of sugar beet is determined significantly by seed vigour and field emergence potential; however, little is known about the molecular mechanisms underlying these traits. Both traits exhibit large variations within sugar beet germplasm that have been difficult to ascribe to either environmental or genetic causes. Among potential targets for trait improvement, an enhancement of stress tolerance is considered because of the high negative influence of environmental stresses on trait parameters. Extending our knowledge of genetic and molecular determinants of sugar beet germination, stress response and adaptation mechanisms would facilitate the detection of new targets for breeding crop with an enhanced field emergence potential. Results To gain insight into the sugar beet germination we initiated an analysis of gene expression in a well emerging sugar beet hybrid showing high germination potential under various environmental conditions. A total of 2,784 ESTs representing 2,251 'unigenes' was generated from dry mature and germinating seeds. Analysis of the temporal expression of these genes during germination under non-stress conditions uncovered drastic transcriptional changes accompanying a shift from quiescent to metabolically active stages of the plant life cycle. Assay of germination under stressful conditions revealed 157 genes showing significantly different expression patterns in response to stress. As deduced from transcriptome data, stress adaptation mechanisms included an alteration in reserve mobilization pathways, an accumulation of the osmoprotectant glycine betaine, late embryogenesis abundant proteins and detoxification enzymes. The observed transcriptional changes are supposed to be regulated by ABA-dependent signal transduction pathway. Conclusion This study

  19. Effects of bagging on sugar metabolism and the activity of sugar ...

    African Journals Online (AJOL)

    To investigate the effects of bagging on sugar metabolism and the activity of sugar metabolism related enzymes in Qingzhong loquat fruit development, the contents of sucrose, glucose and soluble solids as well as the activities of sugar metabolism related enzymes were evaluated. The content of sucrose, glucose and ...

  20. Effects of education on low-phosphate diet and phosphate binder intake to control serum phosphate among maintenance hemodialysis patients: A randomized controlled trial

    Directory of Open Access Journals (Sweden)

    Eunsoo Lim

    2018-03-01

    Full Text Available Background : For phosphate control, patient education is essential due to the limited clearance of phosphate by dialysis. However, well-designed randomized controlled trials about dietary and phosphate binder education have been scarce. Methods : We enrolled maintenance hemodialysis patients and randomized them into an education group (n = 48 or a control group (n = 22. We assessed the patients' drug compliance and their knowledge about the phosphate binder using a questionnaire. Results : The primary goal was to increase the number of patients who reached a calcium-phosphorus product of lower than 55. In the education group, 36 (75.0% patients achieved the primary goal, as compared with 16 (72.7% in the control group (P = 0.430. The education increased the proportion of patients who properly took the phosphate binder (22.9% vs. 3.5%, P = 0.087, but not to statistical significance. Education did not affect the amount of dietary phosphate intake per body weight (education vs. control: -1.18 ± 3.54 vs. -0.88 ± 2.04 mg/kg, P = 0.851. However, the dietary phosphate-to-protein ratio tended to be lower in the education group (-0.64 ± 2.04 vs. 0.65 ± 3.55, P = 0.193. The education on phosphate restriction affected neither the Patient-Generated Subjective Global Assessment score (0.17 ± 4.58 vs. -0.86 ± 3.86, P = 0.363 nor the level of dietary protein intake (-0.03 ± 0.33 vs. -0.09 ± 0.18, P = 0.569. Conclusion : Education did not affect the calcium-phosphate product. Education on the proper timing of phosphate binder intake and the dietary phosphate-to-protein ratio showed marginal efficacy.

  1. Radioactivity of phosphate ores from Karatas-Mazidag phosphate deposit of Turkey

    International Nuclear Information System (INIS)

    Akyuez, T.; Varinlioglu, A.; Kose, A.; Akyuez, S.

    2000-01-01

    The specific activities of 238 U, 226 Ra, 232 Th and 40 K in the composite samples of phosphate ores of type I (grey-coloured ore, with high P 2 O 5 (21-35%) and low calcite content) and of type II (grey coloured calcite ore, with low P 2 O 5 content (5-17%)) of Karatas-Mazidag phosphate deposit, Turkey, have been determined by gamma spectrometry together with phosphatic animal feed ingredients. The concentrations of 238 U, 226 Ra, 232 Th and 40 K were found to be up to 557, 625, 26 and 297 Bq x kg -1 , respectively. Radium equivalent activities of samples were calculated and compared with those given in the literature. Uranium concentration of the individual phosphate samples, from which composite samples of ores of type I and II have been prepared, were found to show and increasing trend with increasing P 2 O 5 and F concentrations. (author)

  2. Monitoring Blood Sugar: The Importance of Checking Blood Sugar Levels

    Science.gov (United States)

    ... closer to the levels seen in people without diabetes) your child's HbA1c, the better controlled the blood sugars have ... Diabetes Hypoglycemia Diabetes Control: Why It's Important Your Child's Diabetes Health Care Team Helping Kids Deal With Injections ...

  3. Performance of several Saccharomyces strains for the alcoholic fermentation of sugar-sweetened high-strength wastewaters: Comparative analysis and kinetic modelling.

    Science.gov (United States)

    Comelli, Raúl N; Seluy, Lisandro G; Isla, Miguel A

    2016-12-25

    This work focuses on the performance of ten commercial Saccharomyces yeast strains in the batch alcoholic fermentation of sugars contained in selected industrial wastewaters from the soft drink industry. Fermentation has been applied successfully to treat these effluents prior to their disposal. Although many strains were investigated, similar behaviour was observed between all of the Saccharomyces strains tested. When media were inoculated with 2gL -1 of yeast, all strains were able to completely consume the available sugars in less than 14h. Thus, any of the strains studied in this work could be used in non-conventional wastewater treatment processes based on alcoholic fermentation. However, ethanol production varied between strains, and these differences could be significant from a production point of view. Saccharomyces bayanus produced the most ethanol, with a mean yield of 0.44g ethanol g sugarconsumed -1 and an ethanol specific production rate of 5.96g ethanol (Lh) -1 . As the assayed soft drinks wastewaters contain about 105g sugar /L of fermentable sugars, the concentration of ethanol achieved after the fermentations process was 46.2g ethanol /L. A rigorous kinetic modelling methodology was used to model the Saccharomyces bayanus fermentation process. The kinetic model included coupled mass balances and a minimal number of parameters. A simple unstructured model based on the Andrews equation (substrate inhibition) was developed. This model satisfactorily described biomass growth, sugar consumption and bioethanol production. In addition to providing insights into the fermentative performance of potentially relevant strains, this work can facilitate the design of large-scale ethanol production processes that use wastewaters from the sugar-sweetened beverage industry as feedstock. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Production of bioethanol from fermented sugars of sugarcane bagasse produced by lignocellulolytic enzymes of Exiguobacterium sp. VSG-1.

    Science.gov (United States)

    Vijayalaxmi, S; Anu Appaiah, K A; Jayalakshmi, S K; Mulimani, V H; Sreeramulu, K

    2013-09-01

    Exiguobacterium sp. VSG-1 was isolated from the soil sample and characterized for the production of lignocellulolytic enzymes. Production of these enzymes by the strain VSG-1 was carried out using steam-exploded sugarcane bagasse (SCB) and found to secrete cellulase, pectinase, mannanase, xylanase, and tannase. The growth and enzyme production were found to be optimum at pH 9.0 and 37 °C. Upon steam explosion of SCB, the cellulose increased by 42 %, whereas hemicelluloses and lignin decreased by 40 and 62 %, respectively. Enzymatic hydrolysis of steam-exploded SCB yielded 640 g/l of total sugars. Fermentation of sugars produced from pretreated SCB was carried out by using Saccharomyces cerevisiae at pH 5.0 and 30 °C. The alcohol produced was calculated and found to be 62.24 g/l corresponding to 78 % of the theoretical yield of ethanol. Hence, the strain VSG-1 has an industrial importance for the production of fermentable sugars for biofuels.

  5. Method for determining the composition of the sugar moiety of a sugar containing compound

    DEFF Research Database (Denmark)

    2016-01-01

    The present invention relates to methods of labeling sugar moieties of sugar containing compounds including glycopeptides. The compounds presented in the present invention facilitate reliable detection of sugar moieties of sugar containing compounds by a combination of spectroscopy methods...

  6. STATISTICAL ANALYSIS OF RAW SUGAR MATERIAL FOR SUGAR PRODUCER COMPLEX

    OpenAIRE

    A. A. Gromkovskii; O. I. Sherstyuk

    2015-01-01

    Summary. In the article examines the statistical data on the development of average weight and average sugar content of sugar beet roots. The successful solution of the problem of forecasting these raw indices is essential for solving problems of sugar producing complex control. In the paper by calculating the autocorrelation function demonstrated that the predominant trend component of the growth raw characteristics. For construct the prediction model is proposed to use an autoregressive fir...

  7. Textual analysis of sugar industry influence on the World Health Organization’s 2015 sugars intake guideline

    Science.gov (United States)

    Reeves, Aaron; Loopstra, Rachel; McKee, Martin

    2016-01-01

    Abstract Objective To determine whether sugar industry-related organizations influenced textual changes between the draft and final versions of the World Health Organization’s (WHO’s) 2015 guideline Sugars intake for adults and children. Methods Stakeholder consultation submissions on the draft guideline from seven sugar industry-related and 10 public health organizations were assessed using the Wordscores program. Document scores were rescaled using the Martin–Vanberg transformation to improve comparability. Draft and final guidelines were compared to identify changes influenced by the sugar industry and public health organizations. Findings There was a small shift in transformed Wordscores score between the draft and final guidelines, from 0.25 to 0.24, towards the industry position. The change was linked to increased use of the word “low” to describe the quality of the evidence, consistent with industry arguments. There was also a shift from use of the word “consumption” to “intake”, irrespective of policy position. Scores for World Sugar Research Organisation and Sugar Nutrition UK submissions ( 0.11 and 0.18, respectively) represented strong pro-industry positions and scores for European Public Health Alliance and Wemos submissions (1.00 and 0.88, respectively) represented the strongest public health positions. Industry tactics included challenging the quality of the evidence, distinguishing between different types of sugar and advocating harm reduction. Conclusion There was little change between draft and final versions of the WHO sugars intake guideline 2015, following industry consultation. The main change was linked to emphasizing the low quality of the evidence on sugar’s adverse effects. Guideline development appeared relatively resistant to industry influence at the stakeholder consultation stage. PMID:27516634

  8. Application of a Colorimetric Assay to Identify Putative Ribofuranosylaminobenzene 5'-Phosphate Synthase Genes Expressed with Activity in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Bechard Matthew E.

    2003-01-01

    Full Text Available Tetrahydromethanopterin (H4MPT is a tetrahydrofolate analog originally discovered in methanogenic archaea, but later found in other archaea and bacteria. The extent to which H4MPT occurs among living organisms is unknown. The key enzyme which distinguishes the biosynthetic pathways of H4MPT and tetrahydrofolate is ribofuranosylaminobenzene 5'-phosphate synthase (RFAP synthase. Given the importance of RFAP synthase in H4MPT biosynthesis, the identification of putative RFAP synthase genes and measurement of RFAP synthase activity would provide an indication of the presence of H4MPT in untested microorganisms. Investigation of putative archaeal RFAP synthase genes has been hampered by the tendency of the resulting proteins to form inactive inclusion bodies in Escherichia coli. The current work describes a colorimetric assay for measuring RFAP synthase activity, and two modified procedures for expressing recombinant RFAP synthase genes to produce soluble, active enzyme. By lowering the incubation temperature during expression, RFAP synthase from Archaeoglobus fulgidus was produced in E. coli and purified to homogeneity. The production of active RFAP synthase from Methanothermobacter thermautotrophicus was achieved by coexpression of the gene MTH0830 with a molecular chaperone. This is the first direct biochemical identification of a methanogen gene that codes for an active RFAP synthase.

  9. Application of a Colorimetric Assay to Identify Putative Ribofuranosylaminobenzene 5'-Phosphate Synthase Genes Expressed with Activity in Escherichia coli.

    Science.gov (United States)

    Bechard, Matthew E.; Chhatwal, Sonya; Garcia, Rosemarie E.; Rasche, Madeline E.

    2003-01-01

    Tetrahydromethanopterin (H(4)MPT) is a tetrahydrofolate analog originally discovered in methanogenic archaea, but later found in other archaea and bacteria. The extent to which H(4)MPT occurs among living organisms is unknown. The key enzyme which distinguishes the biosynthetic pathways of H(4)MPT and tetrahydrofolate is ribofuranosylaminobenzene 5'-phosphate synthase (RFAP synthase). Given the importance of RFAP synthase in H(4)MPT biosynthesis, the identification of putative RFAP synthase genes and measurement of RFAP synthase activity would provide an indication of the presence of H(4)MPT in untested microorganisms. Investigation of putative archaeal RFAP synthase genes has been hampered by the tendency of the resulting proteins to form inactive inclusion bodies in Escherichia coli. The current work describes a colorimetric assay for measuring RFAP synthase activity, and two modified procedures for expressing recombinant RFAP synthase genes to produce soluble, active enzyme. By lowering the incubation temperature during expression, RFAP synthase from Archaeoglobus fulgidus was produced in E. coli and purified to homogeneity. The production of active RFAP synthase from Methanothermobacter thermautotrophicus was achieved by coexpression of the gene MTH0830 with a molecular chaperone. This is the first direct biochemical identification of a methanogen gene that codes for an active RFAP synthase.

  10. Sugar-sweetened Beverage Consumption Among U.S. Adults, 2011-2014.

    Science.gov (United States)

    Rosinger, Asher; Herrick, Kirsten; Gahche, Jaime; Park, Sohyun

    2017-01-01

    Data from the National Health and Nutrition Examination Survey •Approximately one-half of U.S. adults consumed at least one sugar-sweetened beverage on a given day. •Men consumed an average 179 kilocalories (kcal) from sugar-sweetened beverages, which contributed 6.9% of total daily caloric intake. Women consumed an average 113 kcal from sugar-sweetened beverages, which contributed 6.1% of total caloric intake. •Young adults had the highest mean intake and percentage of daily calories from sugar-sweetened beverages relative to older adults. •Non-Hispanic Asian men and women consumed the least calories and the lowest percentage of total calories from sugar-sweetened beverages compared with non-Hispanic white, non-Hispanic black, and Hispanic men and women. Sugar-sweetened beverages are a major contributor of calories and added sugars to diets of U.S. adults (1). Studies have found that sugar-sweetened beverage consumption has been linked to weight gain, metabolic syndrome, dental caries, and type 2 diabetes in adults (2-4). The 2015-2020 Dietary Guidelines for Americans recommend reducing added sugars consumption to less than 10% of total calories per day and, specifically, to choose beverages with no added sugars (1). This report presents results for consumption of sugar-sweetened beverages among U.S. adults aged 20 and over for 2011-2014 by sex, age, and race and Hispanic origin. All material appearing in this report is in the public domain and may be reproduced or copied without permission; citation as to source, however, is appreciated.

  11. Evaluation of intestinal phosphate binding to improve the safety profile of oral sodium phosphate bowel cleansing.

    Directory of Open Access Journals (Sweden)

    Stef Robijn

    Full Text Available Prior to colonoscopy, bowel cleansing is performed for which frequently oral sodium phosphate (OSP is used. OSP results in significant hyperphosphatemia and cases of acute kidney injury (AKI referred to as acute phosphate nephropathy (APN; characterized by nephrocalcinosis are reported after OSP use, which led to a US-FDA warning. To improve the safety profile of OSP, it was evaluated whether the side-effects of OSP could be prevented with intestinal phosphate binders. Hereto a Wistar rat model of APN was developed. OSP administration (2 times 1.2 g phosphate by gavage with a 12h time interval induced bowel cleansing (severe diarrhea and significant hyperphosphatemia (21.79 ± 5.07 mg/dl 6h after the second OSP dose versus 8.44 ± 0.97 mg/dl at baseline. Concomitantly, serum PTH levels increased fivefold and FGF-23 levels showed a threefold increase, while serum calcium levels significantly decreased from 11.29 ± 0.53 mg/dl at baseline to 8.68 ± 0.79 mg/dl after OSP. OSP administration induced weaker NaPi-2a staining along the apical proximal tubular membrane. APN was induced: serum creatinine increased (1.5 times baseline and nephrocalcinosis developed (increased renal calcium and phosphate content and calcium phosphate deposits on Von Kossa stained kidney sections. Intestinal phosphate binding (lanthanum carbonate or aluminum hydroxide was not able to attenuate the OSP induced side-effects. In conclusion, a clinically relevant rat model of APN was developed. Animals showed increased serum phosphate levels similar to those reported in humans and developed APN. No evidence was found for an improved safety profile of OSP by using intestinal phosphate binders.

  12. Dissolvable fluidic time delays for programming multi-step assays in instrument-free paper diagnostics.

    Science.gov (United States)

    Lutz, Barry; Liang, Tinny; Fu, Elain; Ramachandran, Sujatha; Kauffman, Peter; Yager, Paul

    2013-07-21

    Lateral flow tests (LFTs) are an ingenious format for rapid and easy-to-use diagnostics, but they are fundamentally limited to assay chemistries that can be reduced to a single chemical step. In contrast, most laboratory diagnostic assays rely on multiple timed steps carried out by a human or a machine. Here, we use dissolvable sugar applied to paper to create programmable flow delays and present a paper network topology that uses these time delays to program automated multi-step fluidic protocols. Solutions of sucrose at different concentrations (10-70% of saturation) were added to paper strips and dried to create fluidic time delays spanning minutes to nearly an hour. A simple folding card format employing sugar delays was shown to automate a four-step fluidic process initiated by a single user activation step (folding the card); this device was used to perform a signal-amplified sandwich immunoassay for a diagnostic biomarker for malaria. The cards are capable of automating multi-step assay protocols normally used in laboratories, but in a rapid, low-cost, and easy-to-use format.

  13. Design of α-S-Neoglycopeptides Derived from MUC1 with a Flexible and Solvent-Exposed Sugar Moiety

    DEFF Research Database (Denmark)

    Rojas-Ocáriz, Víctor; Compañón, Ismael; Aydillo Miguel, Carlos

    2016-01-01

    in solution have been evaluated by combining NMR experiments and molecular dynamics simulations. The linker plays a key role in the modulation of the conformation of these compounds at different levels, blocking a direct contact between the sugar moiety and the backbone, promoting a helix-like conformation...... for the glycosylated residue and favoring the proper presentation of the sugar unit for molecular recognition events. The feasibility of these novel compounds as mimics of MUC1 antigens has been validated by the X-ray diffraction structure of one of these unnatural derivatives complexed to an anti-MUC1 monoclonal...

  14. Some properties of tetravalent actinide phosphates of M1M42(PO4)3 type and peculiarities of their structure

    International Nuclear Information System (INIS)

    Burnaeva, A.A.; Volkov, Yu.F.; Kryukova, A.I.; Skiba, O.V.; Spiryakov, V.I.; Korshunov, I.A.; Samojlova, T.K.

    1987-01-01

    Generalizing analysis of data on crystallographic and IR spectral properties of double phosphates of actinide and alkali elements of M (1) M 2 (4) (PO 4 ) 3 type is conducted. It is shown, that Li - , Na - , K - , Rb - , CsTh 2 (PO 4 ) 3 , Li - , Na - , KU 2 (PO 4 ) 3 , NaNp 2 (PO 4 ) 3 compounds crystallize in a monoclinic type structure and form an isostructural phosphate series. It is ascertained, that in rubidium-uranium, uranium-cesium systems double salt formation is not observed. Plutonium-sodium phosphate has a rhombic-type structure, which points out to the existence of a morphotropic transition in NaM 2 4 (PO 4 ) 3 phosphate series at the Np-Pu boundary. Some regularities of structure and effect of M 1 and M 4 nature on the double phosphate structure are revealed

  15. Co-ordinate regulation of growth factor receptors and lipid phosphate phosphatase-1 controls cell activation by exogenous lysophosphatidate.

    Science.gov (United States)

    Pilquil, C; Ling, Z C; Singh, I; Buri, K; Zhang, Q X; Brindley, D N

    2001-11-01

    The serum-derived lipid growth factors, lysophosphatidate (LPA) and sphingosine 1-phosphate (S1P), activate cells selectively through different members of a family of endothelial differentiation gene (EDG) receptors. Activation of EDG receptors by LPA and S1P provides a variety of signalling cascades depending upon the G-protein coupling of the different EDG receptors. This leads to chemotactic and mitogenic responses, which are important in wound healing. For example, LPA stimulates fibroblast division and S1P stimulates the chemotaxis and division of endothelial cells leading to angiogenesis. Counteracting these effects of LPA and S1P, are the actions of lipid phosphate phosphatases (LPP, or phosphatidate phosphohydrolases, Type 2). The isoform LPP-1 is expressed in the plasma membrane with its active site outside the cell. This enzyme is responsible for 'ecto-phosphatase' activity leading to the degradation of exogenous lipid phosphate mediators, particularly LPA. Expression of LPP-1 decreases cell activation by exogenous LPA. The mechanism for this is controversial and several mechanisms have been proposed. Evidence will be presented that the LPPs cross-talk with EDG and other growth factor receptors, thus, regulating the responses of the cells to lipid phosphate mediators of signal transduction.

  16. Biomass by-product from crystal sugar production: A comparative study between Ngadirejo and Mauritius sugar mill

    Science.gov (United States)

    Gunawan; Bantacut, T.; Romli, M.; Noor, E.

    2018-03-01

    Sugarcane has been used as raw material in crystal sugar industry. Sugar cane that contains high sugar will be utilized into crystals sugar. In addition, the productivity of sugarcane is large enough in 2016 approximately 360 713 tons/year. Crystal sugar itself is a daily necessity for its use in the food and beverage industry. Problem that occurs in Indonesia is the energy consumption. The sugar mills supposed to be an independent energy source which means it can produce its own energy by utilization the material that is available in the sugar mills such as by-product (bagasse, molasses, filter cake, etc.), the by product in every production stage are quite a lot in sugar industry especially in Indonesia. In this paper, a comparison between two sugar mills was examined between Ngedirejo sugar mill and Mauritius sugar mill which has the same geological state as Indonesia. The results of comparison between the two sugar factories demonstrated the difference in terms of productivity of the sugar that has been produced and the effectiveness of the production process in a sugar mill seen from the amount of waste and the by product.

  17. Isolation and identification of a phosphate solubilising fungus from soil of a phosphate mine in Chaluse, Iran

    Directory of Open Access Journals (Sweden)

    Raheleh Jamshidi

    2016-07-01

    Full Text Available Microbial solubilisation of phosphorus from insoluble phosphates is an environmental friendly and cost effective approach in sustainable soil management. Introducing the indigenous microorganisms to soil requires shorter adaptation period and causes fewer ecological distortions than exogenous microorganisms. This study was conducted to isolate and identify the indigenous fungi for phosphate solubilisation in Mazandaran, Iran. A potent phosphate solubilising fungus was isolated from an Iranian phosphate mine and selected for solubilisation of rock phosphate (RP. The identified fungus was characterised by calmodulin-based polymerase chain reaction method as Aspergillus tubingensis SANRU (Sari Agricultural Sciences and Natural Resources University. The phosphate solubilisation ability of the fungal strain was carried out in shake-flask leaching experiments containing various concentrations of RP (1%, 2%, 4%, or 8% w/v. The maximum P solubilisation rate of 347 mg/l was achieved at 1% of RP concentration on day 9. The regression analysis indicated that the P solubilised mainly through acidification. This study shows the possibility of using A. tubingensis SANRU for application in the management of P fertilisation.

  18. Signal Transduction of Sphingosine-1-Phosphate G Protein—Coupled Receptors

    Directory of Open Access Journals (Sweden)

    Nicholas Young

    2006-01-01

    Full Text Available Sphingosine-1-phosphate (S1P is a bioactive lipid capable of eliciting dramatic effects in a variety of cell types. Signaling by this molecule is by a family of five G protein—coupled receptors named S1P1–5 that signal through a variety of pathways to regulate cell proliferation, migration, cytoskeletal organization, and differentiation. These receptors are expressed in a wide variety of tissues and cell types, and their cellular effects contribute to important biological and pathological functions of S1P in many processes, including angiogenesis, vascular development, lymphocyte trafficking, and cancer. This review will focus on the current progress in the field of S1P receptor signaling and biology.

  19. Early-life exposure to Tris(1,3-dichloroisopropyl) phosphate induces dose-dependent suppression of sexual behavior in male rats.

    Science.gov (United States)

    Kamishima, Manami; Hattori, Tatsuya; Suzuki, Go; Matsukami, Hidenori; Komine, Chiaki; Horii, Yasuyuki; Watanabe, Gen; Oti, Takumi; Sakamoto, Hirotaka; Soga, Tomoko; Parhar, Ishwar S; Kondo, Yasuhiko; Takigami, Hidetaka; Kawaguchi, Maiko

    2018-05-01

    Exposure to endocrine-disrupting chemicals may adversely affect animals, particularly during development. Tris(1,3-dichloroisopropyl) phosphate (TDCIPP) is an organophosphate with anti-androgen function in vitro that is present in indoor dust at relatively high concentrations. In male rats, androgens are necessary for the development of reproductive organs, as well as the endocrine and central nervous systems. However, we currently do not know the exact effects of TDCIPP exposure through suckling on subsequent reproductive behavior in males. Here, we show that TDCIPP exposure (25-250 mg kg -1 via oral administration over 28 consecutive days post-birth) suppressed male sexual behavior and reduced testes size. These changes were dose-dependent and appeared first in adults rather than in juveniles. These results demonstrate that TDCIPP exposure led to normal body growth and appearance in juveniles, but disrupted the endocrine system and physiology in adults. Therefore, assays should be performed using adult animals to ensure accuracy, and to confirm the influence of chemical substances given during early mammalian life. Copyright © 2017 John Wiley & Sons, Ltd.

  20. Erbium-doped phosphate glass waveguide on silicon with 4.1 dB/cm gain at 1.535 µm

    Science.gov (United States)

    Yan, Y. C.; Faber, A. J.; de Waal, H.; Kik, P. G.; Polman, A.

    1997-11-01

    Erbium-doped multicomponent phosphate glass waveguides were deposited by rf sputtering techniques. The Er concentration was 5.3×1020cm-3. By pumping the waveguide at 980 nm with a power of ˜21 mW, a net optical gain of 4.1 dB at 1.535 μm was achieved. This high gain per unit length at low pump power could be achieved because the Er-Er cooperative upconversion interactions in this heavily Er-doped phosphate glass are very weak [the upconversion coefficient is (2.0±0.5)×10-18 cm3/s], presumably due to the homogeneous distribution of Er in the glass and due to the high optical mode confinement in the waveguide which leads to high pump power density at low pump power.

  1. Energy and fructose from beverages sweetened with sugar or high-fructose corn syrup pose a health risk for some people.

    Science.gov (United States)

    Bray, George A

    2013-03-01

    Sugar intake in the United States has increased by >40 fold since the American Revolution. The health concerns that have been raised about the amounts of sugar that are in the current diet, primarily as beverages, are the subject of this review. Just less than 50% of the added sugars (sugar and high-fructose corn syrup) are found in soft drinks and fruit drinks. The intake of soft drinks has increased 5-fold between 1950 and 2000. Most meta-analyses have shown that the risk of obesity, diabetes, cardiovascular disease, and metabolic syndrome are related to consumption of beverages sweetened with sugar or high-fructose corn syrup. Calorically sweetened beverage intake has also been related to the risk of nonalcoholic fatty liver disease, and, in men, gout. Calorically sweetened beverages contribute to obesity through their caloric load, and the intake of beverages does not produce a corresponding reduction in the intake of other food, suggesting that beverage calories are "add-on" calories. The increase in plasma triglyceride concentrations by sugar-sweetened beverages can be attributed to fructose rather than glucose in sugar. Several randomized trials of sugar-containing soft drinks versus low-calorie or calorie-free beverages show that either sugar, 50% of which is fructose, or fructose alone increases triglycerides, body weight, visceral adipose tissue, muscle fat, and liver fat. Fructose is metabolized primarily in the liver. When it is taken up by the liver, ATP decreases rapidly as the phosphate is transferred to fructose in a form that makes it easy to convert to lipid precursors. Fructose intake enhances lipogenesis and the production of uric acid. By worsening blood lipids, contributing to obesity, diabetes, fatty liver, and gout, fructose in the amounts currently consumed is hazardous to the health of some people.

  2. Consumption of Added Sugars and Cardiometabolic Risk Indicators Among US Adolescents

    Science.gov (United States)

    Welsh, Jean A.; Sharma, Andrea; Argeseanu, Solveig; Vos, Miriam B.

    2014-01-01

    Background Increased carbohydrate and sugar consumption has been associated with dyslipidemia among adults. However, the effect of high consumption of added sugars (caloric sweeteners) on measures of cardiometabolic risk among US adolescents is unknown. Methods and Results This was a cross-sectional study of 2,252 US adolescents (13–18 y) in the National Health and Nutrition Examination Survey (NHANES) 1999–2004. Dietary data from one 24-hour recall were merged with added sugar content data from the USDA MyPyramid Equivalents Databases. Multivariate-adjusted means of cardiometabolic indicators were estimated by added sugar consumption level (added sugars was 21.4% of daily energy intake. Adjusted mean high-density lipoprotein cholesterol (HDL) levels were lower, 1.38 mmol/L (95% CI: 1.32, 1.43) among the lowest consumers to 1.28 mmol/L (95% confidence interval [CI]: 1.23, 1.33) among the highest (p-trend=0.007). Geometric mean triglyceride levels ranged from 0.79 mmol/L (95% CI: 0.72, 0.86) to 0.89 mmol/L (95% CI: 0.83, 0.96) (p-trend=0.03) with greater consumption of added sugars. Among those overweight/obese (≥85th percentile body-mass-index [BMI]), HOMA-IRs were positively associated with added sugars (p-linear trendadded sugars is positively associated with measures of cardiometabolic risk. Long-term studies are needed to determine if reduction in added sugars will improve these parameters and, thereby decrease future cardiovascular events. PMID:21220734

  3. Sphingosine-1-Phosphate and the S1P3 Receptor Initiate Neuronal Retraction via RhoA/ROCK Associated with CRMP2 Phosphorylation

    Science.gov (United States)

    Quarta, Serena; Camprubí-Robles, Maria; Schweigreiter, Rüdiger; Matusica, Dusan; Haberberger, Rainer V.; Proia, Richard L.; Bandtlow, Christine E.; Ferrer-Montiel, Antonio; Kress, Michaela

    2017-01-01

    The bioactive lipid sphingosine-1-phosphate (S1P) is an important regulator in the nervous system. Here, we explored the role of S1P and its receptors in vitro and in preclinical models of peripheral nerve regeneration. Adult sensory neurons and motor neuron-like cells were exposed to S1P in an in vitro assay, and virtually all neurons responded with a rapid retraction of neurites and growth cone collapse which were associated with RhoA and ROCK activation. The S1P1 receptor agonist SEW2871 neither activated RhoA or neurite retraction, nor was S1P-induced neurite retraction mitigated in S1P1-deficient neurons. Depletion of S1P3 receptors however resulted in a dramatic inhibition of S1P-induced neurite retraction and was on the contrary associated with a significant elongation of neuronal processes in response to S1P. Opposing responses to S1P could be observed in the same neuron population, where S1P could activate S1P1 receptors to stimulate elongation or S1P3 receptors and retraction. S1P was, for the first time in sensory neurons, linked to the phosphorylation of collapsin response-mediated protein-2 (CRMP2), which was inhibited by ROCK inhibition. The improved sensory recovery after crush injury further supported the relevance of a critical role for S1P and receptors in fine-tuning axonal outgrowth in peripheral neurons. PMID:29066950

  4. Insulin protects apoptotic cardiomyocytes from hypoxia/reoxygenation injury through the sphingosine kinase/sphingosine 1-phosphate axis.

    Directory of Open Access Journals (Sweden)

    Huan Yu

    Full Text Available OBJECTIVE: Experimental and clinical studies have shown that administration of insulin during reperfusion is cardioprotective, but the mechanisms underlying this effect are still unknown. In this study, the ability of insulin to protect apoptotic cardiomyocytes from hypoxia/reoxygenation injury using the sphingosine kinase/sphingosine 1-phosphate axis was investigated. METHODS AND RESULTS: Rat cardiomyocytes were isolated and subjected to hypoxia and reoxygenation. [γ-32P] ATP was used to assess sphingosine kinase activity. Insulin was found to increase sphingosine kinase activity. Immunocytochemistry and Western blot analysis showed changes in the subcellular location of sphingosine kinase 1 from cytosol to the membrane in cardiomyocytes. Insulin caused cardiomyocytes to accumulate of S1P in a dose-dependent manner. FRET efficiency showed that insulin also transactivates the S1P1 receptor. TUNEL staining showed that administration of insulin during reoxygenation could to reduce the rate of reoxygenation-induced apoptosis, which is a requirement for SphK 1 activity. It also reduced the rate of activation of the S1P receptor and inhibited hypoxia/reoxygenation-induced cell death in cardiomyocytes. CONCLUSION: The sphingosine kinase 1/sphingosine 1-phosphate/S1P receptor axis is one pathway through which insulin protects rat cardiomyocytes from apoptosis induced by hypoxia/reoxygenation injury.

  5. The water footprint of sweeteners and bio-ethanol from sugar cane, sugar beet and maize

    NARCIS (Netherlands)

    Gerbens-Leenes, Winnie; Hoekstra, Arjen Ysbert

    2009-01-01

    Sugar cane and sugar beet are used for sugar for human consumption. In the US, maize is used, amongst others, for the sweetener High Fructose Maize Syrup (HFMS). Sugar cane, sugar beet and maize are also important for bio-ethanol production. The growth of crops requires water, a scarce resource. The

  6. Sugar palm (Argena pinnata). Potential of sugar palm for bio-ethanol production

    OpenAIRE

    Elbersen, H.W.; Oyen, L.P.A.

    2010-01-01

    The energetic and economic feasibility of bioethanol production from sugar palm is virtually unknown. A positive factor are the potentially very high yields while the long non-productive juvenile phase and the high labor needs can be seen as problematic. Expansion to large scale sugar palm cultivation comes with risks. Small-scale cultivation of sugar palm perfectly fits into local farming systems. In order to make a proper assessment of the value palm sugar as bio-ethanol crop more informati...

  7. Phosphate steering by Flap Endonuclease 1 promotes 5′-flap specificity and incision to prevent genome instability

    KAUST Repository

    Tsutakawa, Susan E.

    2017-06-27

    DNA replication and repair enzyme Flap Endonuclease 1 (FEN1) is vital for genome integrity, and FEN1 mutations arise in multiple cancers. FEN1 precisely cleaves single-stranded (ss) 5\\'-flaps one nucleotide into duplex (ds) DNA. Yet, how FEN1 selects for but does not incise the ss 5\\'-flap was enigmatic. Here we combine crystallographic, biochemical and genetic analyses to show that two dsDNA binding sites set the 5\\'polarity and to reveal unexpected control of the DNA phosphodiester backbone by electrostatic interactions. Via phosphate steering\\', basic residues energetically steer an inverted ss 5\\'-flap through a gateway over FEN1\\'s active site and shift dsDNA for catalysis. Mutations of these residues cause an 18,000-fold reduction in catalytic rate in vitro and large-scale trinucleotide (GAA) repeat expansions in vivo, implying failed phosphate-steering promotes an unanticipated lagging-strand template-switch mechanism during replication. Thus, phosphate steering is an unappreciated FEN1 function that enforces 5\\'-flap specificity and catalysis, preventing genomic instability.

  8. Anaerobic Treatment of Cane Sugar Effluent from Muhoroni Sugar ...

    African Journals Online (AJOL)

    It was therefore concluded that anaerobic treatment, particularly with pH control and seeding shows potential in first stage management of sugar mill wastewater. Keywords: cane sugar mill effluent, anaerobic treatment, batch reactor, waste stabilization ponds. Journal of Civil Engineering Research and Practice Vol.

  9. Study the Effect of Different Phosphorus Fertilizers on Physiological Characteristic of Photosynthetic Pigments and Soluble Sugars of Safflower under Water Deficit Condition

    Directory of Open Access Journals (Sweden)

    S Heshmati

    2016-10-01

    capacity. The sub-factors were the six treatments resulting from three levels of phosphate chemical fertilizer (0, 50, and 100 kg ha-1 Triple Super Phosphate, each at two levels of Barvar-2 bio-fertilizer (with and without inoculation with Barvar-2. The applied biological fertilizer was in the type of the bacteria which release the phosphorus from soil components and neutralize the soil pH. The commercial name of the biologic fertilizer is Barvar-2. The effective gradients of the biological phosphorus fertilizer is comprised of two bacteria strains of p5 (Bacillus lentus and p13 (Pseudomonas potida with 108 cfu (colony forming units which have been screened from soil bacteria populations. The bacteria strain p5 (Bacillus lentus dissolves P from soil mineral compounds while p13 (Pseudomonas potida separates P from soil organic compounds by exerting a variety of phosphatase enzymes. Results and Discussion The results showed that the highest amount of chlorophyll a, b and total chlorophyll obtained with application of chemical phosphorus by 100 kg ha-1 Triple Super Phosphate followed by inoculation with Barvar-2 under water deficit condition at vegetation and flowering stages. At drought stress in flowering stage, use of high level of phosphorus without inoculation with Barvar-2, had highest effect in terms of increasing amount of chlorophyll a/b ratio, but at treatment of inoculation with Barvar-2, the highest amount of chlorophyll a/ b ratio obtained with low level of chemical phosphorus by 50 kg ha-1 Triple Super Phosphate. The means comparison showed that highest amount of carotenoids, fv/fm, anthocyanin, flavonoids and soluble sugars obtained with application of chemical phosphorus by 50 kg ha-1 Triple Super Phosphate without Barvar-2 under water stress condition at vegetation and flowering stages. The inoculation with Barvar-2 followed by chemical phosphorus by 50 kg ha-1 Triple Super Phosphate significantly increased the amount of anthocyanin and soluble sugars of leaf

  10. Effect of commonly consumed sugar containing and sugar free fruit drinks on the hydrogen ion modulation of human dental plaque

    Directory of Open Access Journals (Sweden)

    Nanika Mahajan

    2014-01-01

    Full Text Available Background: With the increased awareness of healthy diet among the population, the intake of fruit juices as health drinks has been increased. This study has been designed to check the potential cariogenicity of fruit drinks frequently consumed by infants and young children. Aim: To compare the acidogenic potential of sugar free fruit juices with fruit juices containing sugar by evaluating the plaque pH changes, following consumption of the above drinks. Design: The study was carried out on 10 children in the age group of 8-15 years. The four fruit juices used were 1 orange juice with added sugar 2 orange juice with no added sugar 3 apple juice with added sugar 4 apple juice with no added sugar. Sucrose rinse of 10% was used as control group. The endogenous pH of the fruit juices and control was assessed using digital pH meter. The plaque pH was assessed at the baseline and after the consumption of the drinks at 5, 10, 20, 30, 40, and 60 minutes time interval using the plaque-harvesting technique. The obtained results were compiled and subjected to statistical analysis using paired t-test. Result: All the fruit juices showed drop in plaque pH. A drop in pH was also observed in the juices despite of no added sugar content. Conclusion: The fruit juices labeled with "no added sugar" or "free from added sugar", contained substantial quantities of sugar and are equally cariogenic as are fruit drinks with added sugar.

  11. Free Sugar Profile in Cycads

    Directory of Open Access Journals (Sweden)

    Thomas Edward Marler

    2014-10-01

    Full Text Available The sugars fructose, glucose, maltose, and sucrose were quantified in seven tissues of Zamia muricata Willd. to determine their distribution throughout various organs of a model cycad species, and in lateral structural roots of 18 cycad species to determine the variation in sugar concentration and composition among species representing every cycad genus. Taproot and lateral structural roots contained more sugars than leaf, stem, female strobilus, or coralloid roots. For example, taproot sugar concentration was 6.4-fold greater than stem sugar concentration. The dominant root sugars were glucose and fructose, and the only detected stem sugar was sucrose. Sucrose also dominated the sugar profile for leaflet and coralloid root tissue, and fructose was the dominant sugar in female strobilus tissue. Maltose was a minor constituent of taproot, leaflet, and female strobilus tissue, but absent in other tissues. The concentration of total free sugars and each of the four sugars did not differ among genera or families. Stoichiometric relationships among the sugars, such as the quotient hexoses/disaccharides, differed among organs and families. Although anecdotal reports on cycad starch have been abundant due to its historical use as human food and the voluminous medical research invested into cycad neurotoxins, this is the first report on the sugar component of the non-structural carbohydrate profile of cycads. Fructose, glucose, and sucrose are abundant in cycad tissues, with their relative abundance highly contrasting among organs. Their importance as forms of carbon storage, messengers of information, or regulators of cycad metabolism have not been determined to date.

  12. Home blood sugar testing

    Science.gov (United States)

    Diabetes - home glucose testing; Diabetes - home blood sugar testing ... Usual times to test your blood sugar are before meals and at bedtime. Your provider may ask you to check your blood sugar 2 hours after a meal or ...

  13. Sugar from Palms

    DEFF Research Database (Denmark)

    Barfod, Anders

    Throughout the tropics and subtropics a large number of products are derived from the sugar-rich sap tapped from palms. I will give an overview of the most important species being exploited, harvesting practices and yields. I will further provide insights in the biomechanmics of sugar...... transportation in palms, which remain an enigma. Finally, the prospects for developing palm sugar into a commodity of worlswide significance will be discussed....

  14. Hexose-6-phosphate dehydrogenase contributes to skeletal muscle homeostasis independent of 11β-hydroxysteroid dehydrogenase type 1.

    LENUS (Irish Health Repository)

    Semjonous, Nina M

    2011-01-01

    Glucose-6-phosphate (G6P) metabolism by the enzyme hexose-6-phosphate dehydrogenase (H6PDH) within the sarcoplasmic reticulum lumen generates nicotinamide adenine dinucleotide phosphate (reduced) to provide the redox potential for the enzyme 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) to activate glucocorticoid (GC). H6PDH knockout (KO) mice have a switch in 11β-HSD1 activity, resulting in GC inactivation and hypothalamic-pituitary-adrenal axis activation. Importantly, H6PDHKO mice develop a type II fiber myopathy with abnormalities in glucose metabolism and activation of the unfolded protein response (UPR). GCs play important roles in muscle physiology, and therefore, we have examined the importance of 11β-HSD1 and GC metabolism in mediating aspects of the H6PDHKO myopathy. To achieve this, we examined 11β-HSD1\\/H6PDH double-KO (DKO) mice, in which 11β-HSD1 mediated GC inactivation is negated. In contrast to H6PDHKO mice, DKO mice GC metabolism and hypothalamic-pituitary-adrenal axis set point is similar to that observed in 11β-HSD1KO mice. Critically, in contrast to 11β-HSD1KO mice, DKO mice phenocopy the salient features of the H6PDHKO, displaying reduced body mass, muscle atrophy, and vacuolation of type II fiber-rich muscle, fasting hypoglycemia, increased muscle glycogen deposition, and elevated expression of UPR genes. We propose that muscle G6P metabolism through H6PDH may be as important as changes in the redox environment when considering the mechanism underlying the activation of the UPR and the ensuing myopathy in H6PDHKO and DKO mice. These data are consistent with an 11β-HSD1-independent function for H6PDH in which sarcoplasmic reticulum G6P metabolism and nicotinamide adenine dinucleotide phosphate-(oxidized)\\/nicotinamide adenine dinucleotide phosphate (reduced) redox status are important for maintaining muscle homeostasis.

  15. Chemical intervention in plant sugar signalling increases yield and resilience

    Science.gov (United States)

    Griffiths, Cara A.; Sagar, Ram; Geng, Yiqun; Primavesi, Lucia F.; Patel, Mitul K.; Passarelli, Melissa K.; Gilmore, Ian S.; Steven, Rory T.; Bunch, Josephine; Paul, Matthew J.; Davis, Benjamin G.

    2016-12-01

    The pressing global issue of food insecurity due to population growth, diminishing land and variable climate can only be addressed in agriculture by improving both maximum crop yield potential and resilience. Genetic modification is one potential solution, but has yet to achieve worldwide acceptance, particularly for crops such as wheat. Trehalose-6-phosphate (T6P), a central sugar signal in plants, regulates sucrose use and allocation, underpinning crop growth and development. Here we show that application of a chemical intervention strategy directly modulates T6P levels in planta. Plant-permeable analogues of T6P were designed and constructed based on a ‘signalling-precursor’ concept for permeability, ready uptake and sunlight-triggered release of T6P in planta. We show that chemical intervention in a potent sugar signal increases grain yield, whereas application to vegetative tissue improves recovery and resurrection from drought. This technology offers a means to combine increases in yield with crop stress resilience. Given the generality of the T6P pathway in plants and other small-molecule signals in biology, these studies suggest that suitable synthetic exogenous small-molecule signal precursors can be used to directly enhance plant performance and perhaps other organism function.

  16. Sugar Restriction Leads to Increased Ad Libitum Sugar Intake by Overweight Adolescents in an Experimental Test Meal Setting.

    Science.gov (United States)

    O'Reilly, Gillian A; Black, David S; Huh, Jimi; Davis, Jaimie N; Unger, Jennifer; Spruijt-Metz, Donna

    2017-07-01

    The impact of sugar restriction on subsequent sugar intake by overweight adolescents is unknown. Our aim was to examine the effect of sugar restriction on subsequent ad libitum sugar intake by overweight adolescents and whether habitual sugar intake and impulsivity influence the effect of sugar restriction on subsequent sugar intake. This was an in-laboratory crossover feeding trial with sugar-exposure and sugar-restriction conditions. Eighty-seven overweight Latino and African-American adolescents underwent both meal conditions in two separate 8-hour in-laboratory visits. Participants had access to ad libitum snack trays for 3 hours after the condition-specific meals. Ad libitum sugar intake during the snack period was measured at each visit. Habitual sugar intake and impulsivity were assessed at baseline. Repeated measures analysis of covariance was used to examine the within-person effect of meal condition on ad libitum sugar intake. Mixed models were used to examine the moderating effects of habitual sugar intake and impulsivity on the meal condition-ad libitum sugar intake relationship. Participants consumed more ad libitum sugar during the snack period in the sugar-restriction condition than in the sugar-exposure condition (sugar restriction=78.63±38.84 g, sugar exposure=70.86±37.73 g; F=9.64, P=0.002). There was no relationship between habitual sugar intake and how much ad libitum sugar participants consumed during either condition. Higher impulsivity was associated with greater ad libitum sugar intake during both conditions (sugar restriction: b=.029, standard error=.01, Poverweight adolescents restricted from sugar intake consume greater amounts of sugar when they are later given access to high-sugar foods. Overweight adolescents with higher impulsivity appear to consume greater amounts of sugar regardless of previous levels of sugar consumption. Compensatory sugar intake and trait impulsivity may have implications for dietary interventions in this

  17. Serum phosphate and cognitive function in older men.

    Science.gov (United States)

    Slinin, Yelena; Vo, Tien; Taylor, Brent C; Murray, Anne M; Schousboe, John; Langsetmo, Lisa; Ensrud, Kristine

    2018-01-01

    Determine whether serum phosphate is associated with concurrent cognitive impairment and subsequent cognitive decline in older men independent of demographic covariates and atherosclerotic risk factors. In a prospective study of 5529 men enrolled in the Osteoporotic Fractures in Men study, we measured baseline serum phosphate, baseline cognitive function, and change in cognitive function between baseline and follow-up exams an average of 4.6 years later using the Modified Mini-Mental State (3MS) Examination and Trails B. There was no association between serum phosphate and odds of cognitive impairment as assessed by baseline 3MS score or risk of cognitive decline as assessed by longitudinal change in 3MS score. Higher baseline serum phosphate was associated with higher odds of poor executive function as assessed by Trails B with fully adjusted odds ratios 1.12 (95% confidence interval: 0.83-1.52), 1.31 (0.97-1.77), and 1.45 (1.08-1.94) for men in the second, third, and fourth versus the bottom quartile (referent group) of serum phosphate (p-trend 0.007). However, higher phosphate level was not associated with risk of decline in executive function as assessed by longitudinal change in Trails B score with fully adjusted odds ratios 0.94 (95% confidence interval 0.69-1.28), 0.96 (0.70-1.32), and 1.21 (0.89-1.66) for men in the second, third, and fourth versus the bottom quartile (referent group) of serum phosphate (p-trend 0.22). Higher serum phosphate in older men was associated with a higher likelihood of poor executive function, but not with impaired global cognitive function or decline in executive or global cognition. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.

  18. Prerequisite for highly efficient isoprenoid production by cyanobacteria discovered through the over-expression of 1-deoxy-d-xylulose 5-phosphate synthase and carbon allocation analysis.

    Science.gov (United States)

    Kudoh, Kai; Kawano, Yusuke; Hotta, Shingo; Sekine, Midori; Watanabe, Takafumi; Ihara, Masaki

    2014-07-01

    Cyanobacteria have recently been receiving considerable attention owing to their potential as photosynthetic producers of biofuels and biomaterials. Here, we focused on the production of isoprenoids by cyanobacteria, and aimed to provide insight into metabolic engineering design. To this end, we examined the over-expression of a key enzyme in 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway, 1-deoxy-d-xylulose 5-phosphate synthase (DXS) in the cyanobacterium Synechocystis sp. PCC6803. In the DXS-over-expression strain (Dxs_ox), the mRNA and protein levels of DXS were 4-times and 1.5-times the levels in the wild-type (WT) strain, respectively. The carotenoid content of the Dxs_ox strain (8.4 mg/g dry cell weight [DCW]) was also up to 1.5-times higher than that in the WT strain (5.6 mg/g DCW), whereas the glycogen content dramatically decreased to an undetectable level. These observations suggested that the carotenoid content in the Dxs_ox strain was increased by consuming glycogen, which is a C-storage compound in cyanobacteria. We also quantified the total sugar (145 and 104 mg/g DCW), total fatty acids (31 and 24 mg/g DCW) and total protein (200 and 240 mg/g DCW) content in the WT and Dxs_ox strains, respectively, which were much higher than the carotenoid content. In particular, approximately 54% of the proteins were phycobiliproteins. This study demonstrated the major destinations of carbon flux in cyanobacteria, and provided important insights into metabolic engineering. Target yield can be improved through optimization of gene expression, the DXS protein stabilization, cell propagation depression and restriction of storage compound synthesis. Copyright © 2013 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  19. Measurement of biologically active interleukin-1 by a soluble receptor binding assay

    International Nuclear Information System (INIS)

    Riske, F.; Chizzonite, R.; Nunes, P.; Stern, A.S.

    1990-01-01

    A soluble receptor binding assay has been developed for measuring human interleukin-1 alpha (IL-1 alpha), human IL-1 beta, and mouse IL-1 alpha. The assay is based on a competition between unlabeled IL-1 and 125I-labeled mouse recombinant IL-1 alpha for binding to soluble IL-1 receptor prepared from mouse EL-4 cells. The assay measures only biologically active IL-1 folded in its native conformation. The ratio of human IL-1 alpha to human IL-1 beta can be measured in the same sample by a pretreatment step which removes human IL-1 beta from samples prior to assay. This technique has been used to monitor the purification of recombinant IL-1, and may be utilized to specifically and accurately measure bioactive IL-1 in human serum and cell culture supernatants

  20. Preliminary assessment of hydroxyapatite and tricalcium phosphate: in-vitro and in-vivo studies

    International Nuclear Information System (INIS)

    Annuar, B.O.; Muhammad Hasib, A.; Noor Rabihah, A.; Sharifah Anum, Z.; Yaakob, T.A.; Inayat Hussain, S.H.; Rajab, N.F.; Saadiah, S.; Hing, H.L.; Sahidan, S.

    2004-01-01

    Full text: Eight extracts of hydroxyapatite (HA) and tricalcium phosphate (TCP) were evaluated for potential in vitro cytotoxicity, and for dermal irritation and sensitization in vivo. The samples were assessed to determine their viability in L929 murine fibroblast cultures by neutral red (NR) assay, and were evaluated for skin irritation and sensitization in rabbits and guinea pigs, respectively. Results of the NR assay indicate that a majority of extract produced no adverse reaction m L929 cells with cell viability levels exceeding 800/6. However, there was a slight decrease in viability with two HA samples producing a cytotoxicity score of 1. In the animal study, all eight extracts did not promote any dermal irritation and sensitization reactions both in the rabbits and guinea pigs. These results establish that the synthesized HA and TCP am promising biocompatible materials for orthopaedic implants. (Author)

  1. Magnesium prevents phosphate-induced vascular calcification via TRPM7 and Pit-1 in an aortic tissue culture model.

    Science.gov (United States)

    Sonou, Tomohiro; Ohya, Masaki; Yashiro, Mitsuru; Masumoto, Asuka; Nakashima, Yuri; Ito, Teppei; Mima, Toru; Negi, Shigeo; Kimura-Suda, Hiromi; Shigematsu, Takashi

    2017-06-01

    Previous clinical and experimental studies have indicated that magnesium may prevent vascular calcification (VC), but mechanistic characterization has not been reported. This study investigated the influence of increasing magnesium concentrations on VC in a rat aortic tissue culture model. Aortic segments from male Sprague-Dawley rats were incubated in serum-supplemented high-phosphate medium for 10 days. The magnesium concentration in this medium was increased to demonstrate its role in preventing VC, which was assessed by imaging and spectroscopy. The mineral composition of the calcification was analyzed using Fourier transform infrared (FTIR) spectroscopic imaging, scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX) mapping. Magnesium supplementation of high-phosphate medium dose-dependently suppressed VC (quantified as aortic calcium content), and almost ablated it at 2.4 mm magnesium. The FTIR images and SEM-EDX maps indicated that the distribution of phosphate (as hydroxyapatite), phosphorus and Mg corresponded with calcium content in the aortic ring and VC. The inhibitory effect of magnesium supplementation on VC was partially reduced by 2-aminoethoxy-diphenylborate, an inhibitor of TRPM7. Furthermore, phosphate transporter-1 (Pit-1) protein expression was increased in tissues cultured in HP medium and was gradually-and dose dependently-decreased by magnesium. We conclude that a mechanism involving TRPM7 and Pit-1 underpins the magnesium-mediated reversal of high-phosphate-associated VC.

  2. MICROFLORA OF BEET SUGAR PRODUCTION: PROBLEMS AND SOLUTIONS

    Directory of Open Access Journals (Sweden)

    N. G. Kulneva

    2014-01-01

    Full Text Available Summary. Sugar beet is one of the strategic crops for food safety of Russia. The lack of specialized warehouse for harvest does not provide storage of roots for a long time. In the case of a thaw roots that have been defrosted unsuitable for processing. Beet and products of its processing is a good object for the development of microorganisms. Permanent microflora of sugar production are: Bacillus subtilis, Clostridium perfringes, Leuconostoc dextranicum, Torula alba, Pseudomonas fluorescens, Sarcina lutea and other kinds of microorganisms, leading to a problem processing of beet root and reduced quality of sugar. The most dangerous is the slimy bacteriosis is a bacterial disease beet caused by heterofermentative cocci of Leuconostoc (Leuconostoc mesenteroides, L. dextranicum. Product of the vital activity of microorganisms is dextran, which is synthesized from sucrose as a result of dextrany or mucous fermentation and leads to significant technological problems in processing of infected beet. Improving the efficiency of sugar production is connected with decrease in loss of quality of raw material preparation, storing and processing of sugar beet. At sugar plants use a variety of drugs that suppress the growth of pathogenic microflora, but there comes a rapid adaptation of microorganisms, therefore there is a need to implement new products to prevent damage to roots and improve the quality of produced sugar. To resolve this problem experimentally selected bactericidal drug, defined its rational concentration and conditions for the use in sugar beet production. The use of antibacterial drug in the process of extraction allows to increase the purity of diffusion juice 1.3 %, reduce the protein content in it (12.5 %; with the purity of the pure juice increases by 1.1 %, its color index is reduced by 44.7 %.

  3. Complete oxidative conversion of lignocellulose derived non-glucose sugars to sugar acids by Gluconobacter oxydans.

    Science.gov (United States)

    Yao, Ruimiao; Hou, Weiliang; Bao, Jie

    2017-11-01

    Non-glucose sugars derived from lignocellulose cover approximately 40% of the total carbohydrates of lignocellulose biomass. The conversion of the non-glucose sugars to the target products is an important task of lignocellulose biorefining research. Here we report a fast and complete conversion of the total non-glucose sugars from corn stover into the corresponding sugar acids by whole cell catalysis and aerobic fermentation of Gluconobacter oxydans. The conversions include xylose to xylonate, arabinose to arabonate, mannose to mannonate, and galactose to galactonate, as well as with glucose into gluconate. These cellulosic non-glucose sugar acids showed the excellent cement retard setting property. The mixed cellulosic sugar acids could be used as cement retard additives without separation. The conversion of the non-glucose sugars not only makes full use of lignocellulose derived sugars, but also effectively reduces the wastewater treatment burden by removal of residual sugars. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Mechanistic studies of 3-deoxy-D-manno-octulosonic acid 8-phosphate synthase

    International Nuclear Information System (INIS)

    Dotson, G.D.; Woodard, R.W.

    1994-01-01

    The enzyme 3-deOXY-D-manno-octulosonic acid 8-phosphate synthase (KDO 8-P synthase) catalyses the condensation of arabinose 5-phosphate (A 5-P) with phosphoenolpyruvate (PEP) to give the unique eight-carbon acidic sugar 3-deoxy-D-nianno-octulosonic acid 8-phosphate (KDO 8-P) found only in gram-negative bacteria and required for lipid A maturation and cellular growth. The E. coli gene kdsA that encodes KDO 8-P synthase has been amplified by standard PCR methodologies. The synthetic gene, subcloned into the expression vector pT7-7 was used to infect E. coli BL 21 (DE 3). Purification of crude supernatant from this transformant on Q Sepharose yields >200 mg of near-homogeneous KDO 8-P synthase per liter of cell culture. To explore the mechanism of KDO 8-P synthase, we prepared (E)- and (Z)-(3 2 H)PEP, (2- 13 C)PEP, and (2- 13 C, 18 O)PEP chemically from the appropriately labeled 3-bromopyruvates by reaction with trimethylphosphite under Perkow reaction conditions. Our 1 H-NMR analysis of the stereochemistry at C3 of the KDO 8-Ps, obtained by separate incubation of (E)- and (Z)-(3- 2 H)PEP with A 5-P in the presence of KDO 8-P synthase, demonstrated that the reaction is stereospecific with respect to both the C3 of PEP and the C1 carbonyl of A 5-P. (Z)-(3- 2 H)PEP gave predominantly (3S)-(3 2 H)KDO 8-P and (E)-(3- 2 H)PEP gave predominantly (3R)-(3 2 H)KDO-8P, which indicates condensation of the si face of PEP upon the re face of A 5-P-an orientation analogous to that seen with the similar aldehyde Iyase DAH 7-P synthase. The fate of the enolic oxygen of (2- 13 C, 18 O)PEP, during the course of the KDO 8-P synthase-catalyzed reaction as monitored by both 13 C- and 31 P-NMR spectroscopy demonstrated that the inorganic phosphate (Pi) and not the KDO 8-P contained the 18 O

  5. Mechanistic studies of 3-deoxy-D-manno-octulosonic acid 8-phosphate synthase

    Energy Technology Data Exchange (ETDEWEB)

    Dotson, G.D.; Woodard, R.W. [Univ. of Michigan, Ann Arbor, MI (United States)

    1994-12-01

    The enzyme 3-deOXY-D-manno-octulosonic acid 8-phosphate synthase (KDO 8-P synthase) catalyses the condensation of arabinose 5-phosphate (A 5-P) with phosphoenolpyruvate (PEP) to give the unique eight-carbon acidic sugar 3-deoxy-D-nianno-octulosonic acid 8-phosphate (KDO 8-P) found only in gram-negative bacteria and required for lipid A maturation and cellular growth. The E. coli gene kdsA that encodes KDO 8-P synthase has been amplified by standard PCR methodologies. The synthetic gene, subcloned into the expression vector pT7-7 was used to infect E. coli BL 21 (DE 3). Purification of crude supernatant from this transformant on Q Sepharose yields >200 mg of near-homogeneous KDO 8-P synthase per liter of cell culture. To explore the mechanism of KDO 8-P synthase, we prepared (E)- and (Z)-(3{sup 2}H)PEP, (2-{sup 13}C)PEP, and (2-{sup 13}C,{sup 18}O)PEP chemically from the appropriately labeled 3-bromopyruvates by reaction with trimethylphosphite under Perkow reaction conditions. Our {sup 1}H-NMR analysis of the stereochemistry at C3 of the KDO 8-Ps, obtained by separate incubation of (E)- and (Z)-(3-{sup 2}H)PEP with A 5-P in the presence of KDO 8-P synthase, demonstrated that the reaction is stereospecific with respect to both the C3 of PEP and the C1 carbonyl of A 5-P. (Z)-(3-{sup 2}H)PEP gave predominantly (3S)-(3{sup 2}H)KDO 8-P and (E)-(3-{sup 2}H)PEP gave predominantly (3R)-(3{sup 2}H)KDO-8P, which indicates condensation of the si face of PEP upon the re face of A 5-P-an orientation analogous to that seen with the similar aldehyde Iyase DAH 7-P synthase. The fate of the enolic oxygen of (2-{sup 13}C, {sup 18}O)PEP, during the course of the KDO 8-P synthase-catalyzed reaction as monitored by both {sup 13}C- and {sup 31}P-NMR spectroscopy demonstrated that the inorganic phosphate (Pi) and not the KDO 8-P contained the {sup 18}O.

  6. Carbohydrate metabolism in Agaricus bisporus (Lange) Imbach: metabolism of [14C] labelled sugars by sporophores and mycelium

    International Nuclear Information System (INIS)

    Hammond, J.B.W.; Nichols, R.

    1977-01-01

    When growing sporophores and vegetative mycelium of Agaricus bisporus had been supplied with [ 14 C] labelled hexoses for varying periods of time, the major compounds labelled in both forms were mannitol, trehalose, glutamate, alanine and an unidentified substance. Mannitol was strongly labelled after application of [ 14 C] fructose but only weakly when [ 14 C] glucose was applied. The relative rates of oxidation by sporophores of labelled mannitol, trehalose, glucose and fructose were assessed by measuring 14 CO 2 production. Glucose and trehalose were the most rapidly oxidized substrates. Glucosephosphatase isomerase (E.C. 5.3.1.9) was extracted from sporohores and assayed. The conversion of glucose-6-phosphate to fructose-6-phosphate by the enzyme was competitively inhibited in vitro by 6-phosphogluconate. The control of mannitol synthesis and the functions of mannitol and trehalose are discussed. (author)

  7. Respiration-dependent utilization of sugars in yeasts: a determinant role for sugar transporters.

    Science.gov (United States)

    Goffrini, Paola; Ferrero, Iliana; Donnini, Claudia

    2002-01-01

    In many yeast species, including Kluyveromyces lactis, growth on certain sugars (such as galactose, raffinose, and maltose) occurs only under respiratory conditions. If respiration is blocked by inhibitors, mutation, or anaerobiosis, growth does not take place. This apparent dependence on respiration for the utilization of certain sugars has often been suspected to be associated with the mechanism of the sugar uptake step. We hypothesized that in many yeast species, the permease activities for these sugars are not sufficient to ensure the high substrate flow that is necessary for fermentative growth. By introducing additional sugar permease genes, we have obtained K. lactis strains that were capable of growing on galactose and raffinose in the absence of respiration. High dosages of both the permease and maltase genes were indeed necessary for K. lactis cells to grow on maltose in the absence of respiration. These results strongly suggest that the sugar uptake step is the major bottleneck in the fermentative assimilation of certain sugars in K. lactis and probably in many other yeasts.

  8. 76 FR 50285 - Fiscal Year 2012 Tariff-Rate Quota Allocations for Raw Cane Sugar, Refined and Specialty Sugar...

    Science.gov (United States)

    2011-08-12

    ... for Raw Cane Sugar, Refined and Specialty Sugar and Sugar-Containing Products AGENCY: Office of the... quantity of the tariff-rate quotas for imported raw cane sugar, refined and specialty sugar and sugar...), the United States maintains tariff-rate quotas (TRQs) for imports of raw cane sugar and refined sugar...

  9. Oxygen isotope variations in phosphate of biogenic apatites. Pt.1

    International Nuclear Information System (INIS)

    Kolodny, Y.; Luz, B.; Navon, O.

    1983-01-01

    The major advantage of the oxygen in phosphate isotope paleothermometry is that it is a system which records temperatures with great sensitivity while bone (and teeth) building organisms are alive, and the record is nearly perfectly preserved after death. Fish from seven water bodies of different temperatures (3-23 0 C) and different delta 18 O (values - 16 to + 3) of the water were analysed. The delta 18 O values of the analysed PO 4 vary from 6 to 25. The system passed the following tests: (a) the temperature deduced from isotopic analyses of the sequence of fish from Lake Baikal are in good agreement with the temperatures measured in the thermally stratified lake; (b) the isotopic composition of fish bone phosphate is not influenced by the isotopic composition of the phosphate which is fed to the fish, but only by temperature and water composition. Isotopic analysis of fossil fish in combination with analysis of mammal bones should be a useful tool in deciphering continental paleoclimates. (orig.)

  10. Phosphate-enhanced cytotoxicity of zinc oxide nanoparticles and agglomerates.

    Science.gov (United States)

    Everett, W Neil; Chern, Christina; Sun, Dazhi; McMahon, Rebecca E; Zhang, Xi; Chen, Wei-Jung A; Hahn, Mariah S; Sue, H-J

    2014-02-10

    Zinc oxide (ZnO) nanoparticles (NPs) have been found to readily react with phosphate ions to form zinc phosphate (Zn3(PO4)2) crystallites. Because phosphates are ubiquitous in physiological fluids as well as waste water streams, it is important to examine the potential effects that the formation of Zn3(PO4)2 crystallites may have on cell viability. Thus, the cytotoxic response of NIH/3T3 fibroblast cells was assessed following 24h of exposure to ZnO NPs suspended in media with and without the standard phosphate salt supplement. Both particle dosage and size have been shown to impact the cytotoxic effects of ZnO NPs, so doses ranging from 5 to 50 μg/mL were examined and agglomerate size effects were investigated by using the bioinert amphiphilic polymer polyvinylpyrrolidone (PVP) to generate water-soluble ZnO ranging from individually dispersed 4 nm NPs up to micron-sized agglomerates. Cell metabolic activity measures indicated that the presence of phosphate in the suspension media can led to significantly reduced cell viability at all agglomerate sizes and at lower ZnO dosages. In addition, a reduction in cell viability was observed when agglomerate size was decreased, but only in the phosphate-containing media. These metabolic activity results were reflected in separate measures of cell death via the lactate dehydrogenase assay. Our results suggest that, while higher doses of water-soluble ZnO NPs are cytotoxic, the presence of phosphates in the surrounding fluid can lead to significantly elevated levels of cell death at lower ZnO NP doses. Moreover, the extent of this death can potentially be modulated or offset by tuning the agglomerate size. These findings underscore the importance of understanding how nanoscale materials can interact with the components of surrounding fluids so that potential adverse effects of such interactions can be controlled. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  11. Ray tissues as an indirect measure of relative sap-sugar concentration in sugar maple

    Science.gov (United States)

    Peter W. Garrett; Kenneth R. Dudzik; Kenneth R. Dudzik

    1989-01-01

    Attempts to correlate ray tissue as a percentage of total wood volume with sap-sugar concentrations of sugar maple progenies were unsuccessful. These results raise doubts about our ability to use a relatively constant value such as ray-tissue volume in a selection program designed to increase the sap-sugar concentration of sugar maple seedlings.

  12. Application of Calcium Phosphate Materials in Dentistry

    Directory of Open Access Journals (Sweden)

    Jabr S. Al-Sanabani

    2013-01-01

    Full Text Available Calcium phosphate materials are similar to bone in composition and in having bioactive and osteoconductive properties. Calcium phosphate materials in different forms, as cements, composites, and coatings, are used in many medical and dental applications. This paper reviews the applications of these materials in dentistry. It presents a brief history, dental applications, and methods for improving their mechanical properties. Notable research is highlighted regarding (1 application of calcium phosphate into various fields in dentistry; (2 improving mechanical properties of calcium phosphate; (3 biomimetic process and functionally graded materials. This paper deals with most common types of the calcium phosphate materials such as hydroxyapatite and tricalcium phosphate which are currently used in dental and medical fields.

  13. Apolipoprotein M mediates sphingosine-1-phosphate efflux from erythrocytes

    DEFF Research Database (Denmark)

    Christensen, Pernille M.; Bosteen, Markus H.; Hajny, Stefan

    2017-01-01

    Sphingosine-1-phosphate (S1P) is a bioactive lipid implicated in e.g. angiogenesis, lymphocyte trafficking, and endothelial barrier function. Erythrocytes are a main source of plasma S1P together with platelets and endothelial cells. Apolipoprotein M (apoM) in HDL carries 70% of plasma S1P, whereas...... 30% is carried by albumin. The current aim was to investigate the role of apoM in export of S1P from human erythrocytes. Erythrocytes exported S1P more efficiently to HDL than to albumin, particularly when apoM was present in HDL. In contrast, export of sphingosine to HDL was unaffected...... by the presence of apoM. The specific ability of apoM to promote export of S1P was independent of apoM being bound in HDL particles. Treatment with MK-571, an inhibitor of the ABCC1 transporter, effectively reduced export of S1P from human erythrocytes to apoM, whereas the export was unaffected by inhibitors...

  14. Overexpression, crystallization and preliminary X-ray analysis of xylulose-5-phosphate/fructose-6-phosphate phosphoketolase from Bifidobacterium breve

    International Nuclear Information System (INIS)

    Suzuki, Ryuichiro; Kim, Byung-Jun; Shibata, Tsuyoshi; Iwamoto, Yuki; Katayama, Takane; Ashida, Hisashi; Wakagi, Takayoshi; Shoun, Hirofumi; Fushinobu, Shinya; Yamamoto, Kenji

    2010-01-01

    Xylulose-5-phosphate/fructose-6-phosphate phosphoketolase from B. breve was overexpressed and crystallized. The crystals belonged to the tetragonal space group I422 and diffracted to beyond 1.7 Å resolution. The xylulose-5-phosphate/fructose-6-phosphate phosphoketolase gene from Bifidobacterium breve was cloned and overexpressed in Escherichia coli. The enzyme was purified to homogeneity and crystallized by the sitting-drop vapour-diffusion method. Crystals were obtained at 293 K using 0.05 mM thiamine diphosphate, 0.25 mM MgCl 2 , 24%(w/v) PEG 6000 and 0.1 M Bicine pH 9.0. The crystals belonged to the tetragonal space group I422, with unit-cell parameters a = b = 174.8, c = 163.8 Å, and diffracted to beyond 1.7 Å resolution

  15. Biochemical changes correlated with blood thiamine and its phosphate esters levels in patients with diabetes type 1 (DMT1).

    Science.gov (United States)

    Al-Daghri, Nasser M; Alharbi, Mohammed; Wani, Kaiser; Abd-Alrahman, Sherif H; Sheshah, Eman; Alokail, Majed S

    2015-01-01

    Thiamine (vitamin B1) is an essential enzyme cofactor in most organisms required at several stages of anabolic and catabolic intermediary metabolism. However, little is known on the positive effects of thiamine in diabetic type 1 (DMT1) patients. The objectives of this study were to evaluate the biochemical changes related to thiamine deficiency in patients with DMT1 outcomes among Saudi adults. We hypothesized that blood thiamine deficiency in patients with DMT1 manifestations might lead to an increase in metabolic syndrome. A total of 77 patients with DMT1 (age 35.8 ± 5.5) and 81 controls (age 45.0 ± 18.1) (total N = 158) were randomly selected from the Riyadh Cohort Study for inclusion. Saudi adults with diabetes type 1, a significant decrease in systolic (P < 0.001), and diastolic blood pressure (P = 0.008) and microalbuminuria (P = 0.02). Moreover, cholesterol, glucose and triglycerides were significantly increased (P 0.001, 0.001 and 0.008, respectively) in patients with diabetes type 1 compared to controls. On the other hand, HDL, TMP, TDP and thiamine, were significantly decreased in patients with diabetes type 1 (P 0.005, 0.002, 0.005, and 0.002), respectively. A strong association between blood thiamine level and diabetes type 1 was detected in our study population. The results confirmed the role of thiamine and thiamine phosphate esters, in preventing metabolic changes and complications of diabetes type 1. The levels of these thiamine and thiamine phosphate esters were correlated with diabetes related biomarkers including HDL, glucose, triglycerides and cholesterol, as well as microalbuminuria, LDL and urine thiamine. The results support a pivotal role of blood thiamine and its phosphate esters in preventing the biochemical changes and complications in patients with DMT1.

  16. 77 FR 57180 - Fiscal Year 2013 Tariff-rate Quota Allocations for Raw Cane Sugar, Refined and Specialty Sugar...

    Science.gov (United States)

    2012-09-17

    ... Sugar, Refined and Specialty Sugar, and Sugar-Containing Products AGENCY: Office of the United States... quantity of the tariff-rate quotas for imported raw cane sugar, refined and specialty sugar, and sugar... imports of raw cane sugar and refined sugar. Pursuant to Additional U.S. Note 8 to Chapter 17 of the HTS...

  17. Assay and heterologous expression in Pichia pastoris of plant cell wall type-II membrane anchored glycosyltransferases

    DEFF Research Database (Denmark)

    Petersen, Bent; Egelund, Jack; Damager, Iben

    2009-01-01

    .011 to 0.013 U (1 U = 1 nmol conversion of substrate * min(-1) * microl medium(-1)) similar to those of RGXT1 and RGXT2 expressed in Baculovirus transfected insect Sf9 cells. In summary, the data presented suggest that Pichia is an attractive host candidate for expression of plant glycosyltransferases.......Two Arabidopsis xylosyltransferases, designated RGXT1 and RGXT2, were recently expressed in Baculovirus transfected insect cells and by use of the free sugar assay shown to catalyse transfer of D-xylose from UDP-alpha-D-xylose to L-fucose and derivatives hereof. We have now examined expression...

  18. Sonochemically synthesized biocompatible zirconium phosphate nanoparticles for pH sensitive drug delivery application

    Energy Technology Data Exchange (ETDEWEB)

    Kalita, Himani, E-mail: hkalita74@gmail.com [Department of Chemistry, Indian Institute of Technology Kharagpur, West Bengal 721302 (India); Prashanth Kumar, B.N., E-mail: prasanthkumar999@gmail.com [School of Medical Science and Technology, Indian Institute of Technology Kharagpur, West Bengal 721302 (India); Konar, Suraj, E-mail: suraj.konar@gmail.com [Department of Chemistry, Indian Institute of Technology Kharagpur, West Bengal 721302 (India); Tantubay, Sangeeta, E-mail: sang.chem2@gmail.com [Department of Chemistry, Indian Institute of Technology Kharagpur, West Bengal 721302 (India); Mahto, Madhusudan Kr., E-mail: mahtomk0@gmail.com [Department of Chemistry, Indian Institute of Technology Kharagpur, West Bengal 721302 (India); Mandal, Mahitosh, E-mail: mahitosh@smst.iitkgp.ernet.in [School of Medical Science and Technology, Indian Institute of Technology Kharagpur, West Bengal 721302 (India); Pathak, Amita, E-mail: ami@chem.iitkgp.ernet.in [Department of Chemistry, Indian Institute of Technology Kharagpur, West Bengal 721302 (India)

    2016-03-01

    The present work reports the synthesis of biocompatible zirconium phosphate (ZP) nanoparticles as nanocarrier for drug delivery application. The ZP nanoparticles were synthesized via a simple sonochemical method in the presence of cetyltrimethylammonium bromide and their efficacy for the delivery of drugs has been tested through various in-vitro experiments. The particle size and BET surface area of the nanoparticles were found to be ~ 48 nm and 206.51 m{sup 2}/g respectively. The conventional MTT assay and cellular localization studies of the particles, performed on MDA-MB-231 cell lines, demonstrate their excellent biocompatibility and cellular internalization behavior. The loading of curcumin, an antitumor drug, onto the ZP nanoparticles shows the rapid drug uptake ability of the particles, while the drug release study, performed at two different pH values (at 7.4 and 5) depicts pH sensitive release-profile. The MTT assay and cellular localization studies revealed higher cellular inhibition and better bioavailability of the nanoformulated curcumin compared to free curcumin. - Highlights: • Biocompatible zirconium phosphate nanoparticles were synthesized by a simple sonochemical approach. • Curcumin was rapidly loaded onto the particles by the aid by hydrogen bond formation. • The curcumin loaded zirconium phosphate nanoparticles depict pH triggered drug release phenomenon. • The nanoformulated curcumin showed enhanced anti-tumor activity as compared to the native curcumin.

  19. Sonochemically synthesized biocompatible zirconium phosphate nanoparticles for pH sensitive drug delivery application

    International Nuclear Information System (INIS)

    Kalita, Himani; Prashanth Kumar, B.N.; Konar, Suraj; Tantubay, Sangeeta; Mahto, Madhusudan Kr.; Mandal, Mahitosh; Pathak, Amita

    2016-01-01

    The present work reports the synthesis of biocompatible zirconium phosphate (ZP) nanoparticles as nanocarrier for drug delivery application. The ZP nanoparticles were synthesized via a simple sonochemical method in the presence of cetyltrimethylammonium bromide and their efficacy for the delivery of drugs has been tested through various in-vitro experiments. The particle size and BET surface area of the nanoparticles were found to be ~ 48 nm and 206.51 m"2/g respectively. The conventional MTT assay and cellular localization studies of the particles, performed on MDA-MB-231 cell lines, demonstrate their excellent biocompatibility and cellular internalization behavior. The loading of curcumin, an antitumor drug, onto the ZP nanoparticles shows the rapid drug uptake ability of the particles, while the drug release study, performed at two different pH values (at 7.4 and 5) depicts pH sensitive release-profile. The MTT assay and cellular localization studies revealed higher cellular inhibition and better bioavailability of the nanoformulated curcumin compared to free curcumin. - Highlights: • Biocompatible zirconium phosphate nanoparticles were synthesized by a simple sonochemical approach. • Curcumin was rapidly loaded onto the particles by the aid by hydrogen bond formation. • The curcumin loaded zirconium phosphate nanoparticles depict pH triggered drug release phenomenon. • The nanoformulated curcumin showed enhanced anti-tumor activity as compared to the native curcumin.

  20. Apparatus for drying sugar cubes

    NARCIS (Netherlands)

    Derckx, H.A.J.; Torringa, H.M.

    1999-01-01

    Device for drying sugar cubes containing a heating apparatus for heating and dehumidifying the sugar cubes, a conditioning apparatus for cooling off and possibly further dehumidifying the sugar cubes and a conveying apparatus for conveying the sugar cubes through the heating apparatus and the

  1. Changes in the sugar content of sweet sorghum stems under natural conditions during winter in saline soil of the Yellow River Delta

    Science.gov (United States)

    Li, Ying; Yuan, Fang; Wang, Baoshan

    2018-02-01

    In order to investigate the maximum storage period during their natural growth state, the sweet sorghum (Sorghum bicolor L. Moench) stems of four cultivars were analyzed to determine changes in contents of water, total sugars, main soluble sugars and the enzyme activity. From early November 2016 to late January 2017, the decrease in the total sugar content and the contents of sucrose, glucose and fructose slowed down, and the enzyme activities (sucrose synthase and sucrose phosphate synthase) involving sucrose metabolism in the stem remained stable. However, these indicators decreased significantly after the end of January 2017. Low temperatures and a dry environment were conducive to the storage of the sweet sorghum stems. During the winter (from early November 2016 to late January 2017) in northern China, the sweet sorghum plants can be stored naturally in the field via regulating sowing dates, which saves a lot of storage space and production costs for bioethanol company.

  2. Recombinant expression of a functional myo-inositol-1-phosphate synthase (MIPS) in Mycobacterium smegmatis.

    Science.gov (United States)

    Huang, Xinyi; Hernick, Marcy

    2015-10-01

    Myo-inositol-1-phosphate synthase (MIPS, E.C. 5.5.1.4) catalyzes the first step in inositol production-the conversion of glucose-6-phosphate (Glc-6P) to myo-inositol-1-phosphate. While the three dimensional structure of MIPS from Mycobacterium tuberculosis has been solved, biochemical studies examining the in vitro activity have not been reported to date. Herein we report the in vitro activity of mycobacterial MIPS expressed in E. coli and Mycobacterium smegmatis. Recombinant expression in E. coli yields a soluble protein capable of binding the NAD(+) cofactor; however, it has no significant activity with the Glc-6P substrate. In contrast, recombinant expression in M. smegmatis mc(2)4517 yields a functionally active protein. Examination of structural data suggests that MtMIPS expressed in E. coli adopts a fold that is missing a key helix containing two critical (conserved) Lys side chains, which likely explains the inability of the E. coli expressed protein to bind and turnover the Glc-6P substrate. Recombinant expression in M. smegmatis may yield a protein that adopts a fold in which this key helix is formed enabling proper positioning of important side chains, thereby allowing for Glc-6P substrate binding and turnover. Detailed mechanistic studies may be feasible following optimization of the recombinant MIPS expression protocol in M. smegmatis.

  3. Comparison of three PCR-based assays for SNP genotyping in sugar beet

    Science.gov (United States)

    Background: PCR allelic discrimination technologies have broad applications in the detection of single nucleotide polymorphisms (SNPs) in genetics and genomics. The use of fluorescence-tagged probes is the leading method for targeted SNP detection, but assay costs and error rates could be improved t...

  4. The Sugar Tax in Holland

    NARCIS (Netherlands)

    Ajjaji, Fadoua

    2016-01-01

    This inquiry supports the theory of a sugar tax has a positive influence on the sugar consumption of Dutch individuals. Once a tax is implemented, the sugar consumption declines. Furthermore, this study supported the hypothesis claiming that children have a positive influence on their parental sugar

  5. Cytotoxicity and Apoptotic Effects of Polyphenols from Sugar Beet Molasses on Colon Carcinoma Cells in Vitro

    Directory of Open Access Journals (Sweden)

    Mingshun Chen

    2016-06-01

    Full Text Available Three polyphenols were isolated and purified from sugar beet molasses by ultrasonic-aid extraction and various chromatographic techniques, and their structures were elucidated by spectral analysis. Cytotoxicity and the molecular mechanism were measured by methyl thiazolyl tetrazolium (MTT assay, flow cytometry, caspase-3 activity assay and Western blot assay. The results showed that gallic acid, cyanidin-3-O-glucoside chloride and epicatechin have cytotoxicity to the human colon, hepatocellular and breast cancer cells. Cyanidin-3-O-glucoside chloride showed its cytotoxicity against various tumor cell lines, particularly against colon cancer Caco-2 cells with half maximal inhibitory concentration (IC50 value of 23.21 ± 0.14 μg/mL in vitro. Cyanidin-3-O-glucoside chloride may be a potential candidate for the treatment of colon cancer. In the mechanism study, cyanidin-3-O-glucoside chloride increased the ratio of cell cycle at G0/G1 phase and reduced cyclin D1 expression on Caco-2 cells. Cyanidin-3-O-glucoside chloride decreased mutant p21 expression, and increased the ratio of Bax/Bcl-2 and the activation of caspase-3 to induce apoptosis.

  6. The Effect of Ultrasonic Waves on Sugar Extraction and Mechanical Properties of Sugar Beet

    Directory of Open Access Journals (Sweden)

    K Hedayati

    2013-09-01

    Full Text Available Sugar, which can be extracted from sugar cane and sugar beet, is one of the most important ingredients of food. Conducting more research to increase the extraction efficiency of sugar is necessary due to high production of sugar beet and its numerous processing units in northern Khorasan province. In this research, the effect of temperature, time and the frequency of ultrasonic waves on mechanical properties of sugar beet and its extraction rate of sugar in moisture content of 75% were studied. In this regard, an ultrasonic bath in laboratory scale was used. The studied parameters and their levels were frequency in three levels (zero, 25 and 45 KHz, temperature in three levels (25, 50 and 70 ° C and the imposed time of ultrasonic waves in three levels (10, 20 and 30 min. Samples were prepared using planned experiments and the results were compared with control sugar beet samples. A Saccharimeter was used to measure the concenteration of sugar in samples. Two different types of probe including semi-spherical end and the other one with sharpened edges were used to measure mechanical properties. The studied parameters of frequency, temperature and time showed significant effect on sugar extraction and their resulted effect in optimized levels revealed up to 56% increase in sugar extraction compared with control samples. The obtained values of elastic modulus and shear modulus showed a decreasing trend. The obtained values of total energy of rupture, the total energy of shear, the maximum force of rupture, and the yield point of rupture showed an increasing trend. The frequency had no significant effect on the yield point of rupture and shear force.

  7. [Comparison of the clinical performance of the ECLusys HIV combi assay with the Lumipulse f and HISCL 2000-i HIV-1/2 ab screening assays].

    Science.gov (United States)

    Sugiura, Aya; Iwahara, Kunihiro; Suga, Yasuyuki; Uchiyama, Sachinori; Maekawa, Masato

    2012-04-01

    We compared the ECLusys HIV combi assay (ECL HIV Ag/Ab) to the Lumipulse Forte (LPf HIV 1/2 Ab) and HISCL (HIS HIV 1/2 Ab) assays. In a dilution sensitivity test using dilution panels of WHO HIV antibody international reference panel (HIV-1 Subtype A, B, C, E, HIV-1 Group O, HIV-2) and HIV-1/2 Ab CE marked material(HIV-1, HIV-2) parent specimens, the ECL assay enabled detection at a higher level of sensitivity than either the LPf assay or the HIS assay for all dilution panels. In an early detection test in the early phase of infection in which a BBI HIV seroconversion panel was used, the ECL assay enabled detection 7 days after initial blood sample collection, whereas the LPf and HIS assays enabled detection after 27 days. In a specificity test using high RF positive specimens (n=33), pregnancy specimens (n=35), cytomegalovirus antibody positive specimens (n=36), and high M protein positive specimens (n=21) that were confirmed negative for HIV-1/2 antibodies by the LPf assay, negative results were obtained for all specimens on both the ECL assay and the HIS assay. In a correlation test using routinely collected clinical specimens (n=121), including positive stock specimens, the ECL and HIS assays demonstrated the highest agreement rate 98.3%. The above results confirmed that the fourth-generation reagent ECL assay, which simultaneously detects both HIV-1/2 antibodies and p24 antigens, is both highly sensitive and specific, and is a suitable assay for use in routine testing.

  8. Sugar - a harmless indulgence?

    DEFF Research Database (Denmark)

    Rasmussen, Lone Banke; Andersen, Niels Lyhne; Ovesen, L.

    1998-01-01

    The consumption of sugar is relatively high in Denmark - and other industrial countries - and many persons have a consumption which exceeds the recommended level of maximally 10% of energy intake. A high sugar consumption may reduce the nutrient density of the diet and increase the risk of vitamin...... and mineral deficiency, especially in low energy consumers. The sugar intake and the fat intake, expressed as percentage of energy, usually show an inverse association. This has lead to the statement that a diet with both a low sugar content and a low fat content is incompatible, but we will argue...... that this is not the fact. The significance of sugar for the development of obesity is not clarified. A high fat content in the diet seems to promote the development of obesity, while a high carbohydrate content tends to reduce obesity. It is not known if sugar in this connection is comparable to the other carbohydrates...

  9. Nuclear analytical techniques in Cuban sugar industry

    International Nuclear Information System (INIS)

    Diaz R, O.; Griffith M, J.

    1997-01-01

    This paper is a review concerning the application of Nuclear Analytical Techniques in the Cuban sugar industry. The most complete elemental composition of final molasses (34 elements) and natural zeolites (38) this last one employed as an auxiliary agent in sugar technological processe4s has been performed by means of instrumental Neutron Activation Analysis (INAA) and X-Ray Fluorescence Analysis (XRFA). The trace elemental sugar cane soill-plant relationship and elemental composition of different types of Cuban sugar (raw, blanco-directo and refine) were also studied. As a result, valuable information referred to the possibilities of using these products in animal and human foodstuff so as in the other applications are given. (author). 34 refs., 6 figs., 1 tab

  10. Hematopoietic Sphingosine 1-Phosphate Lyase Deficiency Decreases Atherosclerotic Lesion Development in LDL-Receptor Deficient Mice

    NARCIS (Netherlands)

    Bot, Martine; Van Veldhoven, Paul P.; de Jager, Saskia C. A.; Johnson, Jason; Nijstad, Niels; Van Santbrink, Peter J.; Westra, Marijke M.; Van Der Hoeven, Gerd; Gijbels, Marion J.; Mueller-Tidow, Carsten; Varga, Georg; Tietge, Uwe J. F.; Kuiper, Johan; Van Berkel, Theo J. C.; Nofer, Jerzy-Roch; Bot, Ilze; Biessen, Erik A. L.

    2013-01-01

    Aims: Altered sphingosine 1-phosphate (S1P) homeostasis and signaling is implicated in various inflammatory diseases including atherosclerosis. As S1P levels are tightly controlled by S1P lyase, we investigated the impact of hematopoietic S1P lyase (Sgpl1(-/-)) deficiency on leukocyte subsets

  11. Hematopoietic sphingosine 1-phosphate lyase deficiency decreases atherosclerotic lesion development in LDL-receptor deficient mice

    NARCIS (Netherlands)

    Bot, Martine; van Veldhoven, Paul P.; de Jager, Saskia C. A.; Johnson, Jason; Nijstad, Niels; van Santbrink, Peter J.; Westra, Marijke M.; van der Hoeven, Gerd; Gijbels, Marion J.; Müller-Tidow, Carsten; Varga, Georg; Tietge, Uwe J. F.; Kuiper, Johan; van Berkel, Theo J. C.; Nofer, Jerzy-Roch; Bot, Ilze; Biessen, Erik A. L.

    2013-01-01

    Altered sphingosine 1-phosphate (S1P) homeostasis and signaling is implicated in various inflammatory diseases including atherosclerosis. As S1P levels are tightly controlled by S1P lyase, we investigated the impact of hematopoietic S1P lyase (Sgpl1(-/-)) deficiency on leukocyte subsets relevant to

  12. Calcium Overload Accelerates Phosphate-Induced Vascular Calcification Via Pit-1, but not the Calcium-Sensing Receptor.

    Science.gov (United States)

    Masumoto, Asuka; Sonou, Tomohiro; Ohya, Masaki; Yashiro, Mitsuru; Nakashima, Yuri; Okuda, Kouji; Iwashita, Yuko; Mima, Toru; Negi, Shigeo; Shigematsu, Takashi

    2017-07-01

    Vascular calcification (VC) is a risk factor of cardiovascular and all-cause mortality in patients with chronic kidney disease (CKD). CKD-mineral and bone metabolism disorder is an important problem in patients with renal failure. Abnormal levels of serum phosphate and calcium affect CKD-mineral and bone metabolism disorder and contribute to bone disease, VC, and cardiovascular disease. Hypercalcemia is a contributing factor in progression of VC in patients with CKD. However, the mechanisms of how calcium promotes intracellular calcification are still unclear. This study aimed to examine the mechanisms underlying calcium-induced calcification in a rat aortic tissue culture model. Aortic segments from 7-week-old male Sprague-Dawley rats were cultured in serum-supplemented medium for 10 days. We added high calcium (HiCa; calcium 3.0 mM) to high phosphate (HPi; phosphate 3.8 mM) medium to accelerate phosphate and calcium-induced VC. We used phosphonoformic acid and the calcimimetic R-568 to determine whether the mechanism of calcification involves Pit-1 or the calcium-sensing receptor. Medial VC was significantly augmented by HPi+HiCa medium compared with HPi alone (300%, p<0.05), and was associated with upregulation of Pit-1 protein. Pit-1 protein concentrations in HPi+HiCa medium were greater than those in HPi medium. Phosphonoformic acid completely negated the augmentation of medial VC induced by HPi+HiCa. R-568 had no additive direct effect on medial VC. These results indicated that exposure to HPi+HiCa accelerates medial VC, and this is mediated through Pit-1, not the calcium-sensing receptor.

  13. Bioethanol production from Scenedesmus obliquus sugars: the influence of photobioreactors and culture conditions on biomass production.

    Science.gov (United States)

    Miranda, J R; Passarinho, P C; Gouveia, L

    2012-10-01

    A closed-loop vertical tubular photobioreactor (PBR), specially designed to operate under conditions of scarce flat land availability and irregular solar irradiance conditions, was used to study the potential of Scenedesmus obliquus biomass/sugar production. The results obtained were compared to those from an open-raceway pond and a closed-bubble column. The influence of the type of light source and the regime (natural vs artificial and continuous vs light/dark cycles) on the growth of the microalga and the extent of the sugar accumulation was studied in both PBRs. The best type of reactor studied was a closed-loop PBR illuminated with natural light/dark cycles. In all the cases, the relationship between the nitrate depletion and the sugar accumulation was observed. The microalga Scenedesmus was cultivated for 53 days in a raceway pond (4,500 L) and accumulated a maximum sugar content of 29 % g/g. It was pre-treated for carrying out ethanol fermentation assays, and the highest ethanol concentration obtained in the hydrolysate fermented by Kluyveromyces marxianus was 11.7 g/L.

  14. Bioethanol production from Scenedesmus obliquus sugars. The influence of photobioreactors and culture conditions on biomass production

    Energy Technology Data Exchange (ETDEWEB)

    Miranda, J.R.; Passarinho, P.C.; Gouveia, L. [Laboratorio Nacional de Energia e Geologia (LNEG), Lisbon (Portugal). Unidade de Bioenergia

    2012-10-15

    A closed-loop vertical tubular photobioreactor (PBR), specially designed to operate under conditions of scarce flat land availability and irregular solar irradiance conditions, was used to study the potential of Scenedesmus obliquus biomass/sugar production. The results obtained were compared to those from an open-raceway pond and a closed-bubble column. The influence of the type of light source and the regime (natural vs artificial and continuous vs light/dark cycles) on the growth of the microalga and the extent of the sugar accumulation was studied in both PBRs. The best type of reactor studied was a closed-loop PBR illuminated with natural light/dark cycles. In all the cases, the relationship between the nitrate depletion and the sugar accumulation was observed. The microalga Scenedesmus was cultivated for 53 days in a raceway pond (4,500 L) and accumulated a maximum sugar content of 29 % g/g. It was pre-treated for carrying out ethanol fermentation assays, and the highest ethanol concentration obtained in the hydrolysate fermented by Kluyveromyces marxianus was 11.7 g/L. (orig.)

  15. [Production of sugar syrup containing rare sugar using dual-enzyme coupled reaction system].

    Science.gov (United States)

    Han, Wenjia; Zhu, Yueming; Bai, Wei; Izumori, Ken; Zhang, Tongcun; Sun, Yuanxia

    2014-01-01

    Enzymatic conversion is very important to produce functional rare sugars, but the conversion rate of single enzymes is generally low. To increase the conversion rate, a dual-enzyme coupled reaction system was developed. Dual-enzyme coupled reaction system was constructed using D-psicose-3-epimerase (DPE) and L-rhamnose isomerase (L-RhI), and used to convert D-fructose to D-psicose and D-allose. The ratio of DPE and L-RhI was 1:10 (W/W), and the concentration of DPE was 0.05 mg/mL. The optimum temperature was 60 degrees C and pH was 9.0. When the concentration of D-fructose was 2%, the reaction reached its equilibrium after 10 h, and the yield of D-psicose and D-allose was 5.12 and 2.04 g/L, respectively. Using the dual-enzymes coupled system developed in the current study, we could obtain sugar syrup containing functional rare sugar from fructose-rich raw material, such as high fructose corn syrup.

  16. 21 CFR 582.5697 - Riboflavin-5-phosphate.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Riboflavin-5-phosphate. 582.5697 Section 582.5697 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5697 Riboflavin-5-phosphate. (a) Product. Riboflavin-5-phosphate. (b) Conditions of use...

  17. Antimicrobial effects of silver zeolite, silver zirconium phosphate silicate and silver zirconium phosphate against oral microorganisms

    Institute of Scientific and Technical Information of China (English)

    Sirikamon Saengmee-anupharb; Toemsak Srikhirin; Boonyanit Thaweboon; Sroisiri Thaweboon; Taweechai Amornsakchai; Surachai Dechkunakorn; Theeralaksna Suddhasthira

    2013-01-01

    Objective: To evaluate the antimicrobial activities of silver inorganic materials, including silver zeolite (AgZ), silver zirconium phosphate silicate (AgZrPSi) and silver zirconium phosphate (AgZrP), against oral microorganisms. In line with this objective, the morphology and structure of each type of silver based powders were also investigated. Methods: The antimicrobial activities of AgZ, AgZrPSi and AgZrP were tested against Streptococcus mutans, Lactobacillus casei, Candidaalbicans and Staphylococcus aureus using disk diffusion assay as a screening test. The minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) were determined using the modified membrane method. Scanning electron microscope and X-ray diffraction were used to investigate the morphology and structure of these silver materials. Results: All forms of silver inorganic materials could inhibit the growth of all test microorganisms. The MIC of AgZ, AgZrPSi and AgZrP was 10.0 g/L whereas MLC ranged between 10.0-60.0 g/L. In terms of morphology and structure, AgZrPSi and AgZrP had smaller sized particles (1.5-3.0 µm) and more uniformly shaped than AgZ. Conclusions: Silver inorganic materials in the form of AgZ, AgZrPSi and AgZrP had antimicrobial effects against all test oral microorganisms and those activities may be influenced by the crystal structure of carriers. These results suggest that these silver materials may be useful metals applied to oral hygiene products to provide antimicrobial activity against oral infection.

  18. Antimicrobial effects of silver zeolite,silver zirconium phosphate silicate and silver zirconium phosphate against oral microorganisms

    Institute of Scientific and Technical Information of China (English)

    Sirikamon; Saengmee-anupharb; Toemsak; Srikhirin; Boonyanit; Thaweboon; Sroisiri; Thaweboon; Taweechai; Amornsakchai; Surachai; Dechkunakorn; Theeralaksna; Suddhasthira

    2013-01-01

    Objective:To evaluate the antimicrobial activities of silver inorganic materials,including silver zeolite(AgZ),silver zirconium phosphate silicate(AgZrPSi)and silver zirconium phosphate(AgZrp),against oral microorganisms.In line with this objective,the morphology and structure of each type of silver based powders were also investigated.Methods:The antimicrobial activities of AgZ,AgZrPSi and AgZrP were tested against Streptococcus mutans,Lactobacillus casei,Candida albicans and Staphylococcus aureus using disk diffusion assay as a screening test.The minimum inhibitory concentration(MIC)and minimum lethal concentration(MLC)were determined using the modified membrane method.Scanning electron microscope and X-ray diffraction were used to investigate the morphology and structure of these silver materials.Results:All forms of silver inorganic materials could inhibit the growth of all test microorganisms.The MIC of AgZ,AgZrPSi and AgZrP was 10.0 g/L whereas MLC ranged between 10.0-60.0 g/L.In terms of morphology and structure.AgZrPSi and AgZrP had smaller sized particles(1.5-3.0μm)and more uniformly shaped than AgZ.Conclusions:Silver inorganic materials in the form of AgZ,AgZrPSi and AgZrP had antimicrobial effects against all test oral microorganisms and those activities may be influenced by the crystal structure of carriers.These results suggest that these silver materials may be useful metals applied to oral hygiene products to provide antimicrobial activity against oral infection.

  19. myo-Inositol-1-phosphate synthase is required for polar auxin transport and organ development

    KAUST Repository

    Chen, Hao

    2010-06-01

    myo-Inositol-1-phosphate synthase is a conserved enzyme that catalyzes the first committed and rate-limiting step in inositol biosynthesis. Despite its wide occurrence in all eukaryotes, the role of myo-inositol-1-phosphate synthase and de novo inositol biosynthesis in cell signaling and organism development has been unclear. In this study, we isolated loss-of-function mutants in the Arabidopsis MIPS1 gene from different ecotypes. It was found that all mips1 mutants are defective in embryogenesis, cotyledon venation patterning, root growth, and root cap development. The mutant roots are also agravitropic and have reduced basipetal auxin transport. mips1 mutants have significantly reduced levels of major phosphatidylinositols and exhibit much slower rates of endocytosis. Treatment with brefeldin A induces slower PIN2 protein aggregation in mips1, indicating altered PIN2 trafficking. Our results demonstrate that MIPS1 is critical for maintaining phosphatidylinositol levels and affects pattern formation in plants likely through regulation of auxin distribution. © 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

  20. Nodding syndrome in Tanzania may not be associated with circulating anti-NMDA-and anti-VGKC receptor antibodies or decreased pyridoxal phosphate serum levels-a pilot study.

    Science.gov (United States)

    Dietmann, Anelia; Wallner, Bernd; König, Rebekka; Friedrich, Katrin; Pfausler, Bettina; Deisenhammer, Florian; Griesmacher, Andrea; Seger, Christoph; Matuja, William; JilekAall, Louise; Winkler, Andrea S; Schmutzhard, Erich

    2014-06-01

    Nodding syndrome (NS) is a seemingly progressive epilepsy disorder of unknown underlying cause. We investigated association of pyridoxal-phosphate serum levels and occurrence of anti-neuronal antibodies against N-methyl-D-aspartate (NMDA) receptor and voltage gated potassium channel (VGKC) complex in NS patients. Sera of a Tanzanian cohort of epilepsy and NS patients and community controls were tested for the presence of anti-NMDA-receptor and anti-VGKC complex antibodies by indirect immunofluorescence assay. Furthermore pyridoxal-phosphate levels were measured. Auto-antibodies against NMDA receptor or VGKC (LG1 or Caspr2) complex were not detected in sera of patients suffering from NS (n=6), NS plus other seizure types (n=16), primary generalized epilepsy (n=1) and community controls without epilepsy (n=7). Median Pyridoxal-phosphate levels in patients with NS compared to patients with primary generalized seizures and community controls were not significantly different. However, these median pyridoxal-phosphate levels are significantly lower compared to the range considered normal in Europeans. In this pilot study NS was not associated with serum anti-NMDA receptor or anti-VGKC complex antibodies and no association to pyridoxal-phosphate serum levels was found.

  1. Fourier transform Raman spectroscopy of synthetic and biological calcium phosphates.

    Science.gov (United States)

    Sauer, G R; Zunic, W B; Durig, J R; Wuthier, R E

    1994-05-01

    Fourier-transform (FT) Raman spectroscopy was used to characterize the organic and mineral components of biological and synthetic calcium phosphate minerals. Raman spectroscopy provides information on biological minerals that is complimentary to more widely used infrared methodologies as some infrared-inactive vibrational modes are Raman-active. The application of FT-Raman technology has, for the first time, enabled the problems of high sample fluorescence and low signal-to-noise that are inherent in calcified tissues to be overcome. Raman spectra of calcium phosphates are dominated by a very strong band near 960 cm-1 that arises from the symmetric stretching mode (v1) of the phosphate group. Other Raman-active phosphate vibrational bands are seen at approximately 1075 (v3), 590 (v4), and 435 cm-1 (v2). Minerals containing acidic phosphate groups show additional vibrational modes. The different calcium phosphate mineral phases can be distinguished from one another by the relative positions and shapes of these bands in the Raman spectra. FT-Raman spectra of nascent, nonmineralized matrix vesicles (MV) show a distinct absence of the phosphate v1 band even though these structures are rich in calcium and phosphate. Similar results were seen with milk casein and synthetic Ca-phosphatidyl-serine-PO4 complexes. Hence, the phosphate and/or acidic phosphate ions in these noncrystalline biological calcium phosphates is in a molecular environment that differs from that in synthetic amorphous calcium phosphate. In MV, the first distinct mineral phase to form contained acidic phosphate bands similar to those seen in octacalcium phosphate. The mineral phase present in fully mineralized MV was much more apatitic, resembling that found in bones and teeth.(ABSTRACT TRUNCATED AT 250 WORDS)

  2. Sugar-sweetened Beverage Consumption Among U.S. Youth, 2011-2014.

    Science.gov (United States)

    Rosinger, Asher; Herrick, Kirsten; Gahche, Jaime; Park, Sohyun

    2017-01-01

    Data from the National Health and Nutrition Examination Survey •Almost two-thirds of boys and girls consumed at least one sugar-sweetened beverage on a given day. •Boys consumed an average 164 kilocalories (kcal) from sugar-sweetened beverages, which contributed 7.3% of total daily caloric intake. Girls consumed an average 121 kcal from sugar-sweetened beverages, which contributed 7.2% of total daily caloric intake. •Among both boys and girls, older youth had the highest mean intake and percentage of daily calories from sugar-sweetened beverages relative to younger children. •Non-Hispanic Asian boys and girls consumed the least calories and the lowest percentage of total calories from sugar-sweetened beverages compared with non-Hispanic white, non-Hispanic black, and Hispanic boys and girls. Sugar-sweetened beverages contribute calories and added sugars to the diets of U.S. children (1). Studies have suggested a link between the consumption of sugar-sweetened beverages and dental caries, weight gain, type 2 diabetes, dyslipidemia, and nonalcoholic fatty liver disease in children (2-6). The 2015-2020 Dietary Guidelines for Americans recommend reducing added sugars consumption to less than 10% of calories per day and, specifically, to choose beverages with no added sugars (1). This report presents results for consumption of sugar-sweetened beverages among U.S. youth aged 2-19 years for 2011-2014 by sex, age, and race and Hispanic origin. All material appearing in this report is in the public domain and may be reproduced or copied without permission; citation as to source, however, is appreciated.

  3. Lysines 72, 80 and 213 and aspartic acid 210 of the Lactococcus lactis LacR repressor are involved in the response to the inducer tagatose-6-phosphate leading to induction of lac operon expression.

    Science.gov (United States)

    van Rooijen, R J; Dechering, K J; Niek, C; Wilmink, J; de Vos, W M

    1993-02-01

    Site-directed mutagenesis of the Lactococcus lactis lacR gene was performed to identify residues in the LacR repressor that are involved in the induction of lacABCDFEGX operon expression by tagatose-6-phosphate. A putative inducer binding domain located near the C-terminus was previously postulated based on homology studies with the Escherichia coli DeoR family of repressors, which all have a phosphorylated sugar as inducer. Residues within this domain and lysine residues that are charge conserved in the DeoR family were changed into alanine or arginine. The production of the LacR mutants K72A, K80A, K80R, D210A, K213A and K213R in the LacR-deficient L.lactis strain NZ3015 resulted in repressed phospho-beta-galactosidase (LacG) activities and decreased growth rates on lactose. Gel mobility shift assays showed that the complex between a DNA fragment carrying the lac operators and LacR mutants K72A, K80A, K213A and D210A did not dissociate in the presence of tagatose-6-phosphate, in contrast to wild type LacR. Other mutations (K62A/K63A, K72R, K73A, K73R, T212A, F214R, R216R and R216K) exhibited no gross effects on inducer response. The results strongly suggest that the lysines at positions 72, 80 and 213 and aspartic acid at position 210 are involved in the induction of lac operon expression by tagatose-6-phosphate.

  4. Nanoparticle-based, organic receptor coupled fluorescent chemosensors for the determination of phosphate

    Energy Technology Data Exchange (ETDEWEB)

    Kaur, Navneet, E-mail: navneetkaur@pu.ac.in [Centre for Nanoscience and Nanotechnology (UIEAST), Panjab University, Chandigarh 160014 (India); Kaur, Simanpreet; Kaur, Amanpreet [Centre for Nanoscience and Nanotechnology (UIEAST), Panjab University, Chandigarh 160014 (India); Saluja, Preeti; Sharma, Hemant [Department of Chemistry, Indian Institute of Technology Ropar, Rupnagar, Punjab 140001 (India); Saini, Anu; Dhariwal, Nisha [Centre for Nanoscience and Nanotechnology (UIEAST), Panjab University, Chandigarh 160014 (India); Singh, Ajnesh; Singh, Narinder [Department of Chemistry, Indian Institute of Technology Ropar, Rupnagar, Punjab 140001 (India)

    2014-01-15

    The sensors have been developed using silver nanoparticles coated with organic ligands and are fully characterized with spectroscopic methods. The energy-dispersive X-ray (EDX) analysis revealed the presence of organic receptors on the surface of metal nanoparticles. These chemosensors were tested against a range of biological and environmentally relevant cations in the HEPES buffered DMSO/H{sub 2}O (8:2, v/v) solvent system. The fluorescence intensity of these chemosensors was quenched upon coordination with open shell metal ions such as Cu{sup 2+}/Fe{sup 3+}. Anion recognition properties of the corresponding metal complexes have been studied and the original fluorescence intensity of sensors was restored upon addition of phosphate (0–20 µM). Thus, a highly selective chemosensor has been devised for the micromolar estimation of phosphate in semi-aqueous medium. -- Highlights: • The silver nanoparticles have been decorated with organic receptors for chemosensor applications. • The sensor properties are developed for the estimation of phosphate anion. • Thus the sensor relies on the cation displacement assay. • The phosphate sensing event displays the “ON–OFF–ON” mode of switching in sensor.

  5. Dietary sources of sugars in adolescents' diet: the HELENA study.

    Science.gov (United States)

    Mesana, M I; Hilbig, A; Androutsos, O; Cuenca-García, M; Dallongeville, J; Huybrechts, I; De Henauw, S; Widhalm, K; Kafatos, A; Nova, E; Marcos, A; González-Gross, M; Molnar, D; Gottrand, F; Moreno, L A

    2018-03-01

    To report dietary sugars consumption and their different types and food sources, in European adolescents. Food consumption data of selected groups were obtained from 1630 adolescents (45.6% males, 12.5-17.5 years) from the HELENA study using two nonconsecutive 24-h recalls. Energy intake, total sugars and free sugars were assessed using the HELENA-DIAT software. Multiple regression analyses were performed adjusting for relevant confounders. Total sugars intake (137.5 g/day) represented 23.6% and free sugars (110.1 g/day), 19% of energy intake. Girls had significantly lower intakes of energy, carbohydrates, total sugars and free sugars. 94% of adolescents had a consumption of free sugars above 10% of total energy intake. The main food contributor to free sugars was 'carbonated, soft and isotonic drinks,' followed by 'non-chocolate confectionary' and 'sugar, honey, jam and syrup.' Older boys and girls had significantly higher intakes of free sugars from 'cakes, pies and biscuits.' Free sugars intake was negatively associated with low socioeconomic status for 'non-chocolate confectionary' and 'sugar, honey and jam' groups; with low maternal educational level for carbonated and 'soft drinks,' 'sugar, honey and jam,' 'cakes and pies' and 'breakfast cereals' groups; and with high paternal educational level for 'carbonated and soft drinks' and 'chocolates' group. The majority (94%) of studied adolescents consumed free sugars above 10% of daily energy intake. Our data indicate a broad variety in foods providing free sugars. Continued efforts are required at different levels to reduce the intake of free sugars, especially in families with a low educational level.

  6. Effect of reagent type on the froth floatation of Sokoto phosphate ore

    Directory of Open Access Journals (Sweden)

    U.A. Hassan

    2016-06-01

    Full Text Available Effect of reagent type on the froth floatation of Sokoto phosphate ore for its beneficiation has been established. The samples of the Sokoto phosphate mineral ore used for the research work were sourced from mining locations in Dange-Shuni, Bodinga, Yabo, Wurno, and Rabbah Local Government Areas of Sokoto State. Size-Assay analysis conducted on scrubbed Sokoto Phosphates nodules revealed that nodules had a size distribution with 80% passing 29.3 mm. Flotation Tests using AERO704 (fatty Acid, Alkyl Hydroxamates, Melamine as collectors (alone or mixed with diesel, MIBC as frother, Calcium Hydroxide and Sulphuric Acid as pH regulators and Dextrin, Sodium Silicate and Aluminium Chloride as depressants produced poor P2O5 separation in the flotation products due to very poor liberation associated with very fine mineral grains. Based on the results obtained, AERO704 Collector gave the best result with aP2O5 recovery pH of 10.

  7. Hematopoietic Sphingosine 1-Phosphate Lyase Deficiency Decreases Atherosclerotic Lesion Development in LDL-Receptor Deficient Mice

    NARCIS (Netherlands)

    Bot, M.; Veldhoven, van P.P.; Jager, de S.C.; Johnson, J.; Nijstad, N.; van, Santbrink P.J.; Westra, M.M.; Hoeven, van der G.; Gijbels, M.J.; Muller-Tidow, C.; Varga, G.; Tietge, U.J.; Kuiper, J.; Berkel, van T.J.; Nofer, J.R.; Bot, I.; Biessen, E.A.

    2013-01-01

    Abstract Aims Altered sphingosine 1-phosphate (S1P) homeostasis and signaling is implicated in various inflammatory diseases including atherosclerosis. As S1P levels are tightly controlled by S1P lyase, we investigated the impact of hematopoietic S1P lyase (Sgpl1−/−) deficiency on leukocyte

  8. Added sugars and sugar-sweetened beverage consumption, dietary carbohydrate index and depression risk in the Seguimiento Universidad de Navarra (SUN) Project.

    Science.gov (United States)

    Sanchez-Villegas, Almudena; Zazpe, Itziar; Santiago, Susana; Perez-Cornago, Aurora; Martinez-Gonzalez, Miguel A; Lahortiga-Ramos, Francisca

    2018-01-01

    The association between added sugars or sugar-sweetened beverage consumption and the risk of depression, as well as the role of carbohydrate quality in depression risk, remains unclear. Among 15 546 Spanish university graduates from the Seguimiento Universidad de Navarra (SUN) prospective cohort study, diet was assessed with a validated 136-item semi-quantitative FFQ at baseline and at 10-year follow-up. Cumulative average consumption of added sugars, sweetened drinks and an overall carbohydrate quality index (CQI) were calculated. A better CQI was associated with higher whole-grain consumption and fibre intake and lower glycaemic index and consumption of solid (instead of liquid) carbohydrates. Clinical diagnoses of depression during follow-up were classified as incident cases. Multivariable time-dependent Cox regression models were used to estimate hazard ratios (HR) of depression according to consumption of added sugars, sweetened drinks and CQI. We observed 769 incident cases of depression. Participants in the highest quartile of added sugars consumption showed a significant increment in the risk of depression (HR=1·35; 95 % CI 1·09, 1·67, P=0·034), whereas those in the highest quartile of CQI (upper quartile of the CQI) showed a relative risk reduction of 30 % compared with those in the lowest quartile of the CQI (HR=0·70; 95 % CI 0·56, 0·88). No significant association between sugar-sweetened beverage consumption and depression risk was found. Higher added sugars and lower quality of carbohydrate consumption were associated with depression risk in the SUN Cohort. Further studies are necessary to confirm the reported results.

  9. 76 FR 36512 - USDA Increases the Domestic Sugar Overall Allotment Quantity, Reassigns Domestic Cane Sugar...

    Science.gov (United States)

    2011-06-22

    ... imports. The OAQ was increased due to an increase in estimated sugar demand since the FY 2011 OAQ was... sugar imports, as required by law. Upon review of the domestic sugarcane processors' sugar marketing allocations relative to their FY 2011 expected raw sugar supplies, CCC determined that all sugarcane...

  10. SLC1 and SLC4 encode partially redundant acyl-coenzyme A 1-acylglycerol-3-phosphate O-acyltransferases of budding yeast

    DEFF Research Database (Denmark)

    Benghezal, Mohammed; Roubaty, Carole; Veepuri, Vijayanath

    2007-01-01

    Phosphatidic acid is the intermediate, from which all glycerophospholipids are synthesized. In yeast, it is generated from lysophosphatidic acid, which is acylated by Slc1p, an sn-2-specific, acyl-coenzyme A-dependent 1-acylglycerol-3-phosphate O-acyltransferase. Deletion of SLC1 is not lethal...

  11. Estrogen influences dolichyl phosphate distribution among glycolipid pools in mouse uteri

    Energy Technology Data Exchange (ETDEWEB)

    Carson, D.D.; Tang, J.; Hu, G.

    1987-03-24

    To determine the role that dolichyl phosphate availability plays in this induction, the authors studied the effects of estrogen priming on the content of dolichyl phosphate and the distribution of dolichyl phosphate among various glycolipids in uteri. Dolichol-linked saccharides were metabolically labeled to equilibrium with either (/sup 3/H)glucosamine or (/sup 3/H)mannose and extracted from primary explants of uterine tissue. The amount of dolichol-linked saccharide was calculated from the specific radioactivity determined for the corresponding sugar nucleotides extracted from the tissues. The major dolichol-linked saccharides identified were mannosylphosphoryldolichol (MPD), oligosaccharylpyrophosphorydolichol (OSL), and N,N'-diacetylchitobiosylpyrophosphoryldolichol (CBL). Estrogen increased the levels of MPD and OSL 4-fold; however, CBL levels did not change. After 3 days of treatment, the levels of these glycolipids were very similar to those in uteri from pregnant mice. The specific activity of GPD synthase was similar under all conditions studied. These studies provide the first determination of the levels of dolichol-linked saccharides in tissues and how these levels change during hormonal induction of glycoprotein assembly. Coupled with earlier studies, the present work demonstrates that among a number of key points of N-linked oligosaccharide assembly and transfer only synthesis of MPD increases coordinately with the increase observed in lipid- and protein-linked oligosaccharide assembly that occurs in vivo in response to estrogen. They suggest that control of MPD levels is an important regulatory aspect of N-linked glycoprotein assembly in this system.

  12. Lectin binding assays for in-process monitoring of sialylation in protein production.

    Science.gov (United States)

    Xu, Weiduan; Chen, Jianmin; Yamasaki, Glenn; Murphy, John E; Mei, Baisong

    2010-07-01

    Many therapeutic proteins require appropriate glycosylation for their biological activities and plasma half life. Coagulation factor VIII (FVIII) is a glycoprotein which has extensive post-translational modification by N-linked glycosylation. The terminal sialic acid in the N-linked glycans of FVIII is required for maximal circulatory half life. The extent of FVIII sialylation can be determined by high pH anion-exchange chromatography coupled with a pulse electrochemical detector (HPAEC-PED), but this requires a large amount of purified protein. Using FVIII as a model, the objective of the present study was to develop assays that enable detection and prediction of sialylation deficiency at an early stage in the process and thus prevent downstream product quality excursions. Lectin ECA (Erythrina Cristagalli) binds to unsialylated Galbeta1-4 GlcNAc and the ECA-binding level (i.e., terminal Gal(beta1-4) exposure) is inversely proportional to the level of sialylation. By using ECA, a cell-based assay was developed to measure the global sialylation profile in FVIII producing cells. To examine the Galbeta1-4 exposure on the FVIII molecule in bioreactor tissue culture fluid (TCF), an ELISA-based ECA-FVIII binding assay was developed. The ECA-binding specificity in both assays was assessed by ECA-specific sugar inhibitors and neuraminidase digestion. The ECA-binding specificity was also independently confirmed by a ST3GAL4 siRNA knockdown experiment. To establish the correlation between Galbeta1-4 exposure and the HPAEC-PED determined FVIII sialylation value, the FVIII containing bioreactor TCF and the purified FVIII samples were tested with ECA ELISA binding assay. The results indicated an inverse correlation between ECA binding and the corresponding HPAEC-PED sialylation value. The ECA-binding assays are cost effective and can be rapidly performed, thereby making them effective for in-process monitoring of protein sialylation.

  13. In situ formation of the amino sugars 1-amino-1-deoxy-fructose and 2-amino-2-deoxy-glucose under Maillard reaction conditions in the absence of ammonia.

    Science.gov (United States)

    Nashalian, Ossanna; Yaylayan, Varoujan A

    2016-04-15

    Replacing amino acids with their binary metal complexes during the Maillard reaction can initiate various processes, including the oxidative degradation of their glucose conjugates, generating 1-amino-1-deoxy-fructose and its derivatives. These reactive amino sugars are not easily accessible under Maillard reaction conditions and are only formed in the presence of ammonia. To explore the generality of this observation and to study in particular the ability of fructose to generate glucosamine, the amino acid-metal complexes were heated in aqueous solutions with three aldohexoses and two ketohexoses at 110°C for 2 h and the dry residues were analysed by ESI/qTOF/MS/MS. All the sugars generated relatively intense ions at [M+H](+) 180 (C6H14NO5); those ions originating from ketohexoses exhibited MS/MS fragmentations identical to glucosamine and those originating form aldohexoses showed ions identical to fructosamine. Furthermore, the amino sugars were found to form fructosazine, react with other sugars and undergo dehydration reactions. Copyright © 2015 Elsevier Ltd. All rights reserved.

  14. The Malus domestica sugar transporter gene family: identifications based on genome and expression profiling related to the accumulation of fruit sugars.

    Science.gov (United States)

    Wei, Xiaoyu; Liu, Fengli; Chen, Cheng; Ma, Fengwang; Li, Mingjun

    2014-01-01

    In plants, sugar transporters are involved not only in long-distance transport, but also in sugar accumulations in sink cells. To identify members of sugar transporter gene families and to analyze their function in fruit sugar accumulation, we conducted a phylogenetic analysis of the Malus domestica genome. Expression profiling was performed with shoot tips, mature leaves, and developed fruit of "Gala" apple. Genes for sugar alcohol [including 17 sorbitol transporters (SOTs)], sucrose, and monosaccharide transporters, plus SWEET genes, were selected as candidates in 31, 9, 50, and 27 loci, respectively, of the genome. The monosaccharide transporter family appears to include five subfamilies (30 MdHTs, 8 MdEDR6s, 5 MdTMTs, 3 MdvGTs, and 4 MdpGLTs). Phylogenetic analysis of the protein sequences indicated that orthologs exist among Malus, Vitis, and Arabidopsis. Investigations of transcripts revealed that 68 candidate transporters are expressed in apple, albeit to different extents. Here, we discuss their possible roles based on the relationship between their levels of expression and sugar concentrations. The high accumulation of fructose in apple fruit is possibly linked to the coordination and cooperation between MdTMT1/2 and MdEDR6. By contrast, these fruits show low MdSWEET4.1 expression and a high flux of fructose produced from sorbitol. Our study provides an exhaustive survey of sugar transporter genes and demonstrates that sugar transporter gene families in M. domestica are comparable to those in other species. Expression profiling of these transporters will likely contribute to improving our understanding of their physiological functions in fruit formation and the development of sweetness properties.

  15. The Malus domestica sugar transporter gene family: identifications based on genome and expression profiling related to the accumulation of fruit sugars

    Directory of Open Access Journals (Sweden)

    Xiaoyu eWei

    2014-11-01

    Full Text Available In plants, sugar transporters are involved not only in long-distance transport, but also in sugar accumulations in sink cells. To identify members of sugar transporter gene families and to analyze their function in fruit sugar accumulation, we conducted a phylogenetic analysis of the Malus domestica genome. Expression profiling was performed with shoot tips, mature leaves, and developed fruit of ‘Gala’ apple. Genes for sugar alcohol (including 17 sorbitol transporters, sucrose, and monosaccharide transporters, plus SWEET genes, were selected as candidates in 31, 9, 50, and 27 loci, respectively, of the genome. The monosaccharide transporter family appears to include five subfamilies (30 MdHTs, 8 MdEDR6s, 5 MdTMTs, 3 MdvGTs, and 4 MdpGLTs. Phylogenetic analysis of the protein sequences indicated that orthologs exist among Malus, Vitis, and Arabidopsis. Investigations of transcripts revealed that 68 candidate transporters are expressed in apple, albeit to different extents. Here, we discuss their possible roles based on the relationship between their levels of expression and sugar concentrations. The high accumulation of fructose in apple fruit is possibly linked to the coordination and cooperation between MdTMT1/2 and MdEDR6. By contrast, these fruits show low MdSWEET4.1 expression and a high flux of fructose produced from sorbitol. Our study provides an exhaustive survey of sugar transporter genes and demonstrates that sugar transporter gene families in M. domestica are comparable to those in other species. Expression profiling of these transporters will likely contribute to improving our understanding of their physiological functions in fruit formation and the development of sweetness properties.

  16. The variable charge of andisols as affected by nanoparticles of rock phosphate and phosphate solubilizing bacteria

    Science.gov (United States)

    Arifin, M.; Nurlaeny, N.; Devnita, R.; Fitriatin, B. N.; Sandrawati, A.; Supriatna, Y.

    2018-02-01

    Andisols has a great potential as agriculture land, however, it has a high phosphorus retention, variable charge characteristics and high value of zero net charge or pH0. The research is aimed to study the effects of nanoparticles of rock phosphate (NPRP) and biofertilizer (phosphate solubilizing bacteria/PSB) on soil pH, pHo (zero point of charge, ZPC) and organic-C in one subgroup of Andisols, namely Acrudoxic Durudands, Ciater Region West Java. The research was conducted from October 2016 to February 2017 in Soil Physics Laboratory and Laboratory of Soil Chemistry and Fertility, Soil Science Department, Faculty of Agriculture, Universitas Padjadjaran. This experiment used a completely randomized factorial design, consisting of two factors and three replications. The first factor was nanoparticles of rock phosphate consist of 4 doses 0; 25; 50 and 75 g/1 kg soil and the second factor was biofertilizer dose consist of g/1 kg soil and without biofertilizer. Total treatment combinations were 8 with 3 replications, so there were 24 experimental plots. The results showed that in general NPRR and biofertilizer will decrease the value of soil pH throughout the incubation periods. There is an interaction between nanoparticles of rock phosphate and biofertilizer in decreasing pHo in the first month of incubation, but after 4-month incubation period, NPRP increased. Interaction between 75 g nanoparticles of rock phosphate with 1 g biofertilizer/1 kg soil in fourth months of incubation decreased soil organic-C to 3.35%.

  17. The signaling lipid sphingosine 1-phosphate regulates mechanical pain

    Science.gov (United States)

    Hill, Rose Z; Hoffman, Benjamin U; Morita, Takeshi; Campos, Stephanie M; Lumpkin, Ellen A; Brem, Rachel B

    2018-01-01

    Somatosensory neurons mediate responses to diverse mechanical stimuli, from innocuous touch to noxious pain. While recent studies have identified distinct populations of A mechanonociceptors (AMs) that are required for mechanical pain, the molecular underpinnings of mechanonociception remain unknown. Here, we show that the bioactive lipid sphingosine 1-phosphate (S1P) and S1P Receptor 3 (S1PR3) are critical regulators of acute mechanonociception. Genetic or pharmacological ablation of S1PR3, or blockade of S1P production, significantly impaired the behavioral response to noxious mechanical stimuli, with no effect on responses to innocuous touch or thermal stimuli. These effects are mediated by fast-conducting A mechanonociceptors, which displayed a significant decrease in mechanosensitivity in S1PR3 mutant mice. We show that S1PR3 signaling tunes mechanonociceptor excitability via modulation of KCNQ2/3 channels. Our findings define a new role for S1PR3 in regulating neuronal excitability and establish the importance of S1P/S1PR3 signaling in the setting of mechanical pain thresholds. PMID:29561262

  18. Epigenetic regulation of pro-inflammatory cytokine secretion by sphingosine 1-phosphate (S1P) in acute lung injury: Role of S1P lyase.

    Science.gov (United States)

    Ebenezer, David L; Fu, Panfeng; Suryadevara, Vidyani; Zhao, Yutong; Natarajan, Viswanathan

    2017-01-01

    Cellular level of sphingosine-1-phosphate (S1P), the simplest bioactive sphingolipid, is tightly regulated by its synthesis catalyzed by sphingosine kinases (SphKs) 1 & 2 and degradation mediated by S1P phosphatases, lipid phosphate phosphatases, and S1P lyase. The pleotropic actions of S1P are attributed to its unique inside-out (extracellular) signaling via G-protein-coupled S1P1-5 receptors, and intracellular receptor independent signaling. Additionally, S1P generated in the nucleus by nuclear SphK2 modulates HDAC1/2 activity, regulates histone acetylation, and transcription of pro-inflammatory genes. Here, we present data on the role of S1P lyase mediated S1P signaling in regulating LPS-induced inflammation in lung endothelium. Blocking S1P lyase expression or activity attenuated LPS-induced histone acetylation and secretion of pro-inflammatory cytokines. Degradation of S1P by S1P lyase generates Δ2-hexadecenal and ethanolamine phosphate and the long-chain fatty aldehyde produced in the cytoplasmic compartment of the endothelial cell seems to modulate histone acetylation pattern, which is different from the nuclear SphK2/S1P signaling and inhibition of HDAC1/2. These in vitro studies suggest that S1P derived long-chain fatty aldehyde may be an epigenetic regulator of pro-inflammatory genes in sepsis-induced lung inflammation. Trapping fatty aldehydes and other short chain aldehydes such as 4-hydroxynonenal derived from S1P degradation and lipid peroxidation, respectively by cell permeable agents such as phloretin or other aldehyde trapping agents may be useful in treating sepsis-induced lung inflammation via modulation of histone acetylation. . Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. A quantitative assay measuring the function of lipase maturation factor 1

    Science.gov (United States)

    Yin, Fen; Doolittle, Mark H.; Péterfy, Miklós

    2009-01-01

    Newly synthesized lipoprotein lipase (LPL) and related members of the lipase gene family require an endoplasmic reticulum maturation factor for attainment of enzyme activity. This factor has been identified as lipase maturation factor 1 (Lmf1), and mutations affecting its function and/or expression result in combined lipase deficiency (cld) and hypertriglyceridemia. To assess the functional impact of Lmf1 sequence variations, both naturally occurring and induced, we report the development of a cell-based assay using LPL activity as a quantitative reporter of Lmf1 function. The assay uses a cell line homozygous for the cld mutation, which renders endogenous Lmf1 nonfunctional. LPL transfected into the mutant cld cell line fails to attain activity; however, cotransfection of LPL with wild-type Lmf1 restores its ability to support normal lipase maturation. In this report, we describe optimized conditions that ensure the detection of a complete range of Lmf1 function (full, partial, or complete loss of function) using LPL activity as the quantitative reporter. To illustrate the dynamic range of the assay, we tested several novel mutations in mouse Lmf1. Our results demonstrate the ability of the assay to detect and analyze Lmf1 mutations having a wide range of effects on Lmf1 function and protein expression. PMID:19471043

  20. 75 FR 53013 - Fiscal Year 2011 Tariff-rate Quota Allocations for Raw Cane Sugar, Refined and Specialty Sugar...

    Science.gov (United States)

    2010-08-30

    ... for Raw Cane Sugar, Refined and Specialty Sugar, and Sugar-containing Products; Revision AGENCY... August 17, 2010 concerning Fiscal Year 2011 tariff-rate quota allocations of raw cane sugar, refined and special sugar, and sugar-containing products. USTR is revising the effective date of that notice to...

  1. A Mos1 transposase in vivo assay to screen new HIV-1 integrase inhibitors.

    Science.gov (United States)

    Cancian, Mariana; Loreto, Elgion L S

    2018-04-01

    The integrase and transposase enzymes of retrovirus and transposons, respectively, share the catalytic DDE domain. In vitro assays showed that inhibitors of HIV-1 integrase generally inhibit the mariner Mos1 transposase. Using a Drosophila strain in which the mobilisation of the mariner element can be quantified by mosaic eyes, we showed that flies maintained in medium containing 210 µM to 4 mM of raltegravir, or 1 or 2 mM of dolutegravir, which are HIV-1 integrase inhibitor used in AIDS treatment, have 23-33% less somatic mobilisation in mosaic eyes when treated with raltegravir and 28-32% when treated with dolutegravir. The gene expression of the mariner transposase gene, estimated by qPCR, is similar among treated and control flies. The results suggest that in vivo assays using Drosophila can be used as a primary screening of inhibitory drugs for transposase and retroviral integrase. The advantages of this assay are that it is easy, quick, cheap and is an in vivo test, meaning that the tested substance has to have been taken in by cells and has arrived at the target site, which is not the case when in vitro assays are applied.

  2. Conditions promoting and restraining agronomic effectiveness of water-insoluble phosphate sources, in particular phosphate rock (PR): I. Indices of phosphate rock use opportunity (PRUOIS) and of phosphate rock suitability for direct use (PRSIDU)

    International Nuclear Information System (INIS)

    Borlan, Z.; Gavriluta, I.; Soare, M.; Stefanescu, D.; Alexandrescu, A.

    2002-01-01

    Several issues of phosphate rock (PR) use are discussed in this paper. Maize for green fodder (Zea mays L) and ryegrass (Lolium multiflorum Lam.) were grown in 7 kg of dry soil and in small pots of 1.25 kg dry soil capacity, respectively, on several base unsaturated soils belonging to Hapludoll and Hapludalf soil groups. The amount of phosphate rock (PR) to apply was based on experimental data considering soil adsorbed acidity (Ah), humus content (H 2 ), cation exchange capacity (T), sum of exchangeable bases (SEB) and mobile (easily soluble) phosphate content (P A L) in the soil. The factors were combined in a rock phosphate use, opportunity index of the soil (PRUOIS): PRUOIS=(A h *H 2 *100)/SEB*10 0.0245*P AL Rock phosphate suitability for direct use was evaluated by means of the rate of PR-P dissolution (PRPRS) in a 0.6% ammonium heptamolybdate in 0.01M calcium chloride solution (ppm P) and by carbonate content (%CaCO 3 ) in PR. Both of these parameters combined provided a phosphate rock suitability index for direct use (PRSIDU): PRSIDU [ppmP/min]=PRPRS*(1-0.03*CaCO 3 ) Water insoluble P sources studied were PR from Kola-Russia, Morocco, Kneifiss-Siria, El Hassa-Jordan, Gafsa- Tunisia, North-Carolina (USA), and Arad-Israel. All PRs were compared with TSP applied at the same rate of P. Neither PRUOIS or PRSIDU considered separately could satisfactorily explain the variance of PR efficiency. An index obtained by multiplicative combination of PRUOIS x PRSIDU did correlate significantly with indices on the agronomic efficiency of PR. (author)

  3. Worldwide trends in dietary sugars intake.

    Science.gov (United States)

    Wittekind, Anna; Walton, Janette

    2014-12-01

    Estimating trends in dietary intake data is integral to informing national nutrition policy and monitoring progress towards dietary guidelines. Dietary intake of sugars is a controversial public health issue and guidance in relation to recommended intakes is particularly inconsistent. Published data relating to trends in sugars intake are relatively sparse. The purpose of the present review was to collate and review data from national nutrition surveys to examine changes and trends in dietary sugars intake. Only thirteen countries (all in the developed world) appear to report estimates of sugars intake from national nutrition surveys at more than one point in time. Definitions of dietary sugars that were used include 'total sugars', 'non-milk extrinsic sugars', 'added sugars', sucrose' and 'mono- and disaccharides'. This variability in terminology across countries meant that comparisons were limited to within countries. Hence trends in dietary sugars intake were examined by country for the whole population (where data permitted), and for specific or combined age and sex subpopulations. Findings indicate that in the majority of population comparisons, estimated dietary sugars intake is either stable or decreasing in both absolute (g/d) and relative (% energy) terms. An increase in sugars intake was observed in few countries and only in specific subpopulations. In conclusion, the findings from the present review suggest that, in the main, dietary sugars intake are decreasing or stable. A consistent approach to estimation of dietary sugars intake from national nutrition surveys is required if more valid estimates of changes in dietary sugars intakes are required in the future.

  4. Native and reconstituted HDL activate Stat3 in ventricular cardiomyocytes via ERK1/2: role of sphingosine-1-phosphate.

    Science.gov (United States)

    Frias, Miguel A; James, Richard W; Gerber-Wicht, Christine; Lang, Ursula

    2009-05-01

    High-density lipoprotein (HDL) has been reported to have cardioprotective properties independent from its cholesterol transport activity. The influence of native HDL and reconstituted HDL (rHDL) on Stat3, the transcription factor playing an important role in myocardium adaptation to stress, was analysed in neonatal rat ventricular cardiomyocytes. We have investigated modulating the composition of rHDL as a means of expanding its function and potential cardioprotective effects. Stat3 phosphorylation and activation were determined by western blotting and electrophoretic mobility shift assay (EMSA). In ventricular cardiomyocytes, HDL and the HDL constituent sphingosine-1-phosphate (S1P) induce a concentration- and time-dependent increase in Stat3 activation. They also enhance extracellular signal-regulated kinases (ERK1/2) and p38 mitogen-activated protein kinase (MAPK) phosphorylation. U0126, a specific inhibitor of MEK1/2, the upstream activator of ERK1/2, abolishes HDL- and S1P-induced Stat3 activation, whereas the p38 MAPK blocker SB203580 has no significant effect. Inhibition of the tyrosine kinase family Src (Src) caused a significant reduction of Stat3 activation, whereas inhibition of phosphatidylinositol 3-kinase (PI3K) had no effect. S1P and rHDL containing S1P have a similar strong stimulatory action on Stat3, ERK1/2, and p38 MAPK comparable to native HDL. S1P-free rHDL has a much weaker effect. Experiments with agonists and antagonists of the S1P receptor subtypes indicate that HDL and S1P activate Stat3 mainly through the S1P2 receptor. In ventricular cardiomyocytes, addition of S1P to rHDL enhances its therapeutic potential by improving its capacity to activate Stat3. Activation of Stat3 occurs mainly via the S1P constituent and the lipid receptor S1P2 requiring stimulation of ERK1/2 and Src but not p38 MAPK or PI3K. The study underlines the therapeutic potential of tailoring rHDL to confront particular clinical situations.

  5. Analysis of Onset Mechanisms of a Sphingosine 1-Phosphate Receptor Modulator Fingolimod-Induced Atrioventricular Conduction Block and QT-Interval Prolongation

    Energy Technology Data Exchange (ETDEWEB)

    Yagi, Yukihiro [Department of Pharmacology, Faculty of Medicine, Toho University, 5-21-16 Omori-nishi, Ota-ku, Tokyo 143–8540 (Japan); Pharmaceutical Research Center, Meiji Seika Pharma Co., Ltd., 760 Morooka-cho, Kohoku-ku, Yokohama, Kanagawa 222–8567 (Japan); Nakamura, Yuji [Department of Pharmacology, Faculty of Medicine, Toho University, 5-21-16 Omori-nishi, Ota-ku, Tokyo 143–8540 (Japan); Kitahara, Ken [Department of Pharmacology, Faculty of Medicine, Toho University, 5-21-16 Omori-nishi, Ota-ku, Tokyo 143–8540 (Japan); Division of Cardiovascular Medicine, Department of Internal Medicine, Faculty of Medicine, Toho University, 6-11-1 Omori-nishi, Ota-ku, Tokyo 143–8541 (Japan); Harada, Takuma [Department of Pharmacology, Faculty of Medicine, Toho University, 5-21-16 Omori-nishi, Ota-ku, Tokyo 143–8540 (Japan); Kato, Kazuhiko; Ninomiya, Tomohisa [Pharmaceutical Research Center, Meiji Seika Pharma Co., Ltd., 760 Morooka-cho, Kohoku-ku, Yokohama, Kanagawa 222–8567 (Japan); Cao, Xin [Department of Pharmacology, Faculty of Medicine, Toho University, 5-21-16 Omori-nishi, Ota-ku, Tokyo 143–8540 (Japan); Ohara, Hiroshi [Department of Pharmacology, Faculty of Medicine, Toho University, 5-21-16 Omori-nishi, Ota-ku, Tokyo 143–8540 (Japan); Division of Cardiovascular Medicine, Department of Internal Medicine, Faculty of Medicine, Toho University, 6-11-1 Omori-nishi, Ota-ku, Tokyo 143–8541 (Japan); Izumi-Nakaseko, Hiroko [Department of Pharmacology, Faculty of Medicine, Toho University, 5-21-16 Omori-nishi, Ota-ku, Tokyo 143–8540 (Japan); Suzuki, Kokichi [Pharmaceutical Research Center, Meiji Seika Pharma Co., Ltd., 760 Morooka-cho, Kohoku-ku, Yokohama, Kanagawa 222–8567 (Japan); Ando, Kentaro [Department of Pharmacology, Faculty of Medicine, Toho University, 5-21-16 Omori-nishi, Ota-ku, Tokyo 143–8540 (Japan); and others

    2014-11-15

    Fingolimod, a sphingosine 1-phosphate (S1P) receptor subtype 1, 3, 4 and 5 modulator, has been used for the treatment of patients with relapsing forms of multiple sclerosis, but atrioventricular conduction block and/or QT-interval prolongation have been reported in some patients after the first dose. In this study, we directly compared the electropharmacological profiles of fingolimod with those of siponimod, a modulator of sphingosine 1-phosphate receptor subtype 1 and 5, using in vivo guinea-pig model and in vitro human ether-a-go-go-related gene (hERG) assay to better understand the onset mechanisms of the clinically observed adverse events. Fingolimod (0.01 and 0.1 mg/kg) or siponimod (0.001 and 0.01 mg/kg) was intravenously infused over 10 min to the halothane-anaesthetized guinea pigs (n = 4), whereas the effects of fingolimod (1 μmol/L) and siponimod (1 μmol/L) on hERG current were examined (n = 3). The high doses of fingolimod and siponimod induced atrioventricular conduction block, whereas the low dose of siponimod prolonged PR interval, which was not observed by that of fingolimod. The high dose of fingolimod prolonged QT interval, which was not observed by either dose of siponimod. Meanwhile, fingolimod significantly inhibited hERG current, which was not observed by siponimod. These results suggest that S1P receptor subtype 1 in the heart could be one of the candidates for fingolimod- and siponimod-induced atrioventricular conduction block since S1P receptor subtype 5 is localized at the brain, and that direct I{sub Kr} inhibition may play a key role in fingolimod-induced QT-interval prolongation. - Highlights: • Fingolimod and siponimod are S1P{sub 1,3,4,5} and S1P{sub 1,5} receptor modulators, respectively. • Fingolimod and siponimod induced AV block in the halothane-anesthetized guinea pigs. • S1P{sub 1} in the hearts may be the target of fingolimod- and siponimod-induced AV block. • Fingolimod directly inhibited hERG current, which was not

  6. Analysis of Onset Mechanisms of a Sphingosine 1-Phosphate Receptor Modulator Fingolimod-Induced Atrioventricular Conduction Block and QT-Interval Prolongation

    International Nuclear Information System (INIS)

    Yagi, Yukihiro; Nakamura, Yuji; Kitahara, Ken; Harada, Takuma; Kato, Kazuhiko; Ninomiya, Tomohisa; Cao, Xin; Ohara, Hiroshi; Izumi-Nakaseko, Hiroko; Suzuki, Kokichi; Ando, Kentaro

    2014-01-01

    Fingolimod, a sphingosine 1-phosphate (S1P) receptor subtype 1, 3, 4 and 5 modulator, has been used for the treatment of patients with relapsing forms of multiple sclerosis, but atrioventricular conduction block and/or QT-interval prolongation have been reported in some patients after the first dose. In this study, we directly compared the electropharmacological profiles of fingolimod with those of siponimod, a modulator of sphingosine 1-phosphate receptor subtype 1 and 5, using in vivo guinea-pig model and in vitro human ether-a-go-go-related gene (hERG) assay to better understand the onset mechanisms of the clinically observed adverse events. Fingolimod (0.01 and 0.1 mg/kg) or siponimod (0.001 and 0.01 mg/kg) was intravenously infused over 10 min to the halothane-anaesthetized guinea pigs (n = 4), whereas the effects of fingolimod (1 μmol/L) and siponimod (1 μmol/L) on hERG current were examined (n = 3). The high doses of fingolimod and siponimod induced atrioventricular conduction block, whereas the low dose of siponimod prolonged PR interval, which was not observed by that of fingolimod. The high dose of fingolimod prolonged QT interval, which was not observed by either dose of siponimod. Meanwhile, fingolimod significantly inhibited hERG current, which was not observed by siponimod. These results suggest that S1P receptor subtype 1 in the heart could be one of the candidates for fingolimod- and siponimod-induced atrioventricular conduction block since S1P receptor subtype 5 is localized at the brain, and that direct I Kr inhibition may play a key role in fingolimod-induced QT-interval prolongation. - Highlights: • Fingolimod and siponimod are S1P 1,3,4,5 and S1P 1,5 receptor modulators, respectively. • Fingolimod and siponimod induced AV block in the halothane-anesthetized guinea pigs. • S1P 1 in the hearts may be the target of fingolimod- and siponimod-induced AV block. • Fingolimod directly inhibited hERG current, which was not observed by

  7. Sugar, Pressure and Pregnancy

    African Journals Online (AJOL)

    2017.v33i1.337. 1. Van den Berghe G, Wouters P, Weekers F, et al. Intensive insulin therapy in critically ill patients. N Engl J Med 2001;345(19):1359-1367. https://doi.org/10.1056/nejmoa011300. 2. The NICE-SUGAR Study Investigators. Intensive ...

  8. Dextransucrase production using cashew apple juice as substrate: effect of phosphate and yeast extract addition.

    Science.gov (United States)

    Chagas, Clarice M A; Honorato, Talita L; Pinto, Gustavo A S; Maia, Geraldo A; Rodrigues, Sueli

    2007-05-01

    Cashew apples are considered agriculture excess in the Brazilian Northeast because cashew trees are cultivated primarily with the aim of cashew nut production. In this work, the use of cashew apple juice as a substrate for Leuconostoc mesenteroides cultivation was investigated. The effect of yeast extract and phosphate addition was evaluated using factorial planning tools. Both phosphate and yeast extract addition were significant factors for biomass growth, but had no significant effect on maximum enzyme activity. The enzyme activities found in cashew apple juice assays were at least 3.5 times higher than the activity found in the synthetic medium. Assays with pH control (pH = 6.5) were also carried out. The pH-controlled fermentation enhanced biomass growth, but decreased the enzyme activity. Crude enzyme free of cells produced using cashew apple juice was stable for 16 h at 30 degrees C at a pH of 5.0.

  9. Thirteen years of observations on primary sugars and sugar alcohols over remote Chichijima Island in the western North Pacific

    Science.gov (United States)

    Verma, Santosh Kumar; Kawamura, Kimitaka; Chen, Jing; Fu, Pingqing

    2018-01-01

    In order to understand the atmospheric transport of bioaerosols, we conducted long-term observations of primary sugars and sugar alcohols over remote Chichijima Island in the western North Pacific from 2001 to 2013. Our results showed that concentrations of total sugar compounds for 13 years ranged from 1.2 to 310 ng m-3 (average of 46 ± 49 ng m-3). We found that atmospheric circulations significantly affect the seasonal variations of bioaerosol distributions over the western North Pacific. The primary sugars (glucose and fructose) maximized in summer, possibly due to an increased emission of the vegetation products from local vascular plants in Chichijima. We also found higher concentrations of sugar components (arabitol, mannitol, and trehalose) in more recent years during summer and autumn, suggesting an enhanced emission of fungal and microbial species over the island. Sucrose peaked in late winter to early spring, indicating a springtime pollen contribution by long-range atmospheric transport, while elevated concentrations of sucrose in early summer could be explained by long-range transport of soil dust from Southeast Asia to Chichijima. Sucrose and trehalose were found to present increasing trends from 2001 to 2013, while total sugar components did not show any clear trends during the 13-year period. Positive matrix factorization analyses suggested the locally emitted sugar compounds as well as long-range-transported airborne pollen grains, microbes, and fungal spores are the major contributors to total sugar compounds in the Chichijima aerosols. Backward air mass trajectories support the atmospheric transport of continental aerosols from the Asian continent during winter and spring over Chichijima.

  10. Effects of polyhexamethylene guanidine phosphate on human gingival fibroblasts.

    Science.gov (United States)

    Vitt, Anton; Slizen, Veronica; Boström, Elisabeth A; Yucel-Lindberg, Tülay; Kats, Anna; Sugars, Rachael V; Gustafsson, Anders; Buhlin, Kåre

    2017-10-01

    Polyhexamethylene guanidine phosphate (PHMG-P) was compared to chlorhexidine (CHX) in order to determine potential cytotoxic and immune-modulatory effects on human gingival fibroblasts. Cytotoxic effects of PHMG-P and CHX on human gingival fibroblasts were assessed using cell viability assay at various time points and concentrations. The effects of PHMG-P and CHX on the secretion of prostaglandin (PG) E 2 , interleukin (IL)-6, IL-8 and matrix metalloproteinase (MMP)-1 by non-stimulated or IL-1β stimulated fibroblasts were evaluated by enzyme-linked immunosorbent assays. PHMG-P concentration 0.00009% led to the total loss of fibroblast viability within 24 h, whereas inhibition of fibroblast viability by CHX occurred at significantly higher concentrations of 0.0009% (p PHMG-P led to loss of fibroblast viability after 5 min, whilst cells exposed to 0.005% CHX survived 30 min of treatment (p PHMG-P or CHX at concentrations of 0.000045 or 0.0.00009% resulted in significantly decreased PGE 2 , IL-6, IL-8 and MMP-1 levels. PHMG-P or CHX alone did not affect the baseline secretion of PGE 2 , IL-6, IL-8 or MMP-1 by gingival fibroblasts. Cytotoxic effects on gingival fibroblasts were triggered by both PHMG-P and CHX at concentrations below those used in clinical practice. The tested antiseptics did not cause inflammation and reduced IL-1β-induced secretion of inflammatory mediators and collagenase by gingival fibroblasts, which suggests anti-inflammatory properties.

  11. Phosphate control in dialysis

    Directory of Open Access Journals (Sweden)

    Cupisti A

    2013-10-01

    Full Text Available Adamasco Cupisti,1 Maurizio Gallieni,2 Maria Antonietta Rizzo,2 Stefania Caria,3 Mario Meola,4 Piergiorgio Bolasco31Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy; 2Nephrology and Dialysis Unit, San Carlo Borromeo Hospital, Milan, Italy; 3Territorial Department of Nephrology and Dialysis, ASL Cagliari, Italy; 4Sant'Anna School of Advanced Studies, University of Pisa, Pisa, ItalyAbstract: Prevention and correction of hyperphosphatemia is a major goal of chronic kidney disease–mineral and bone disorder (CKD–MBD management, achievable through avoidance of a positive phosphate balance. To this aim, optimal dialysis removal, careful use of phosphate binders, and dietary phosphate control are needed to optimize the control of phosphate balance in well-nourished patients on a standard three-times-a-week hemodialysis schedule. Using a mixed diffusive–convective hemodialysis tecniques, and increasing the number and/or the duration of dialysis tecniques are all measures able to enhance phosphorus (P mass removal through dialysis. However, dialytic removal does not equal the high P intake linked to the high dietary protein requirement of dialysis patients; hence, the use of intestinal P binders is mandatory to reduce P net intestinal absorption. Unfortunately, even a large dose of P binders is able to bind approximately 200–300 mg of P on a daily basis, so it is evident that their efficacy is limited in the case of an uncontrolled dietary P load. Hence, limitation of dietary P intake is needed to reach the goal of neutral phosphate balance in dialysis, coupled to an adequate protein intake. To this aim, patients should be informed and educated to avoid foods that are naturally rich in phosphate and also processed food with P-containing preservatives. In addition, patients should preferentially choose food with a low P-to-protein ratio. For example, patients could choose egg white or protein from a vegetable source

  12. Methods for dehydration of sugars and sugar alcohols

    Science.gov (United States)

    Holladay, Johnathan E [Kennewick, WA; Hu, Jianli [Kennewick, WA; Zhang, Xinjie [Burlington, MA; Wang, Yong [Richland, WA

    2010-08-10

    The invention includes a method of dehydration of a sugar using a dehydration catalyst and a co-catalyst within a reactor. A sugar is introduced and H.sub.2 is flowed through the reactor at a pressure of less than or equal to about 300 psig to convert at least some of the sugar into an anhydrosugar product. The invention includes a process for producing isosorbide. A starting material comprising sorbitol is flowed into a reactor. H.sub.2 is counter flowed through the reactor. The starting material is exposed to a catalyst in the presence of a co-catalyst which comprises at least one metal. The exposing is conducted at a hydrogen pressure of less than or equal to 300 psig within the reactor and the hydrogen removes at least some of any water present during the exposing and inhibits formation of colored byproducts.

  13. Stability study for magnetic reagent assaying Hb and HbA1c

    International Nuclear Information System (INIS)

    Hsieh, Wen-Pin; Chieh, J.J.; Yang, C.C.; Yang, S.Y.; Chen, Po-Yu; Huang, Yu-Hao; Hong, Y.W.; Horng, H.E.

    2013-01-01

    Reagents for magnetically labeled immunoassay on human Hb and human HbA1c have been synthesized. The reagents consist of Fe 3 O 4 magnetic particles biofunctionalized with antibodies against Hb and HbA1c. It has been demonstrated that the reagents can be applied to quantitatively detect Hb and HbA1c by using immunomagnetic reduction assay. In addition to characterizing the assay properties, such as the standard curve and the low-detection limit, the stability of reagents is investigated. To do this, the temporal dependence of particle sizes and the bio-activity of reagents are monitored. The results show that the reagents are highly stable when stored at 2–8 °C. This means that the reagents synthesized in this work are promising for practical applications. - Highlights: ► The properties of assaying Hb and HbA1c using immunomagnetic reduction are studied. ► The magnetic nanoparticles with antibodies are highly stable in solutions. ► No significant mutual interference between Hb and HbA1c in assays is observed. ► High-sensitivity assays on Hb and HbA1c using immunomagnetic reduction are achieved.

  14. Ambient aerosol concentrations of sugars and sugar-alcohols at four different sites in Norway

    Directory of Open Access Journals (Sweden)

    K. E. Yttri

    2007-08-01

    Full Text Available Sugars and sugar-alcohols are demonstrated to be important constituents of the ambient aerosol water-soluble organic carbon fraction, and to be tracers for primary biological aerosol particles (PBAP. In the present study, levels of four sugars (fructose, glucose, sucrose, trehalose and three sugar-alcohols (arabitol, inositol, mannitol in ambient aerosols have been quantified using a novel HPLC/HRMS-TOF (High Performance Liquid Chromatography in combination with High Resolution Mass Spectrometry – Time of Flight method to assess the contribution of PBAP to PM>sub>10 and PM2.5. Samples were collected at four sites in Norway at different times of the year in order to reflect the various contributing sources and the spatial and seasonal variation of the selected compounds.

    Sugars and sugar-alcohols were present at all sites investigated, underlining the ubiquity of these highly polar organic compounds. The highest concentrations were reported for sucrose, reaching a maximum concentration of 320 ng m−3 in PM10 and 55 ng m−3 in PM2.5. The mean concentration of sucrose was up to 10 times higher than fructose, glucose and the dimeric sugar trehalose. The mean concentrations of the sugar-alcohols were typically lower, or equal, to that of the monomeric sugars and trehalose. Peak concentrations of arabitol and mannitol did not exceed 30 ng m−3 in PM10, and for PM2.5 all concentrations were below 6 ng m−3.

    Sugars and sugar-alcohols were associated primarily with coarse aerosols except during wintertime at the suburban site in Elverum, where a shift towards sub micron aerosols was observed. It is proposed that this shift was due to the intensive use of wood burning for residential heating at this site during winter, confirmed by high concurrent concentrations of levoglucosan. Elevated concentrations of sugars in PM2

  15. Structural and Functional Insight of Sphingosine 1-Phosphate-Mediated Pathogenic Metabolic Reprogramming in Sickle Cell Disease.

    Science.gov (United States)

    Sun, Kaiqi; D'Alessandro, Angelo; Ahmed, Mostafa H; Zhang, Yujin; Song, Anren; Ko, Tzu-Ping; Nemkov, Travis; Reisz, Julie A; Wu, Hongyu; Adebiyi, Morayo; Peng, Zhangzhe; Gong, Jing; Liu, Hong; Huang, Aji; Wen, Yuan Edward; Wen, Alexander Q; Berka, Vladimir; Bogdanov, Mikhail V; Abdulmalik, Osheiza; Han, Leng; Tsai, Ah-Lim; Idowu, Modupe; Juneja, Harinder S; Kellems, Rodney E; Dowhan, William; Hansen, Kirk C; Safo, Martin K; Xia, Yang

    2017-11-10

    Elevated sphingosine 1-phosphate (S1P) is detrimental in Sickle Cell Disease (SCD), but the mechanistic basis remains obscure. Here, we report that increased erythrocyte S1P binds to deoxygenated sickle Hb (deoxyHbS), facilitates deoxyHbS anchoring to the membrane, induces release of membrane-bound glycolytic enzymes and in turn switches glucose flux towards glycolysis relative to the pentose phosphate pathway (PPP). Suppressed PPP causes compromised glutathione homeostasis and increased oxidative stress, while enhanced glycolysis induces production of 2,3-bisphosphoglycerate (2,3-BPG) and thus increases deoxyHbS polymerization, sickling, hemolysis and disease progression. Functional studies revealed that S1P and 2,3-BPG work synergistically to decrease both HbA and HbS oxygen binding affinity. The crystal structure at 1.9 Å resolution deciphered that S1P binds to the surface of 2,3-BPG-deoxyHbA and causes additional conformation changes to the T-state Hb. Phosphate moiety of the surface bound S1P engages in a highly positive region close to α1-heme while its aliphatic chain snakes along a shallow cavity making hydrophobic interactions in the "switch region", as well as with α2-heme like a molecular "sticky tape" with the last 3-4 carbon atoms sticking out into bulk solvent. Altogether, our findings provide functional and structural bases underlying S1P-mediated pathogenic metabolic reprogramming in SCD and novel therapeutic avenues.

  16. Effect of altered sink:source ratio on photosynthetic metabolism of source leaves

    International Nuclear Information System (INIS)

    Plaut, Z.; Mayoral, M.L.; Reinhold, L.

    1987-01-01

    When seven crop species were grown under identical environmental conditions, decreased sink:source ratio led to a decreased photosynthetic rate within 1 to 3 days in Cucumis sativus L., Gossypium hirsutum L., and Raphanus sativus L., but not in Capsicum annuum L., Solanum melongena L., Phaseolus vulgaris L., or Ricinus communis L. The decrease was not associated with stomatal closure. In cotton and cucumbers, sink removal led to an increase in starch and sugar content, in glucose 6-phosphate and fructose 6-phosphate pools, and in the proportion of 14 C detected in sugar phosphates and UDPglucose following 14 CO 2 supply. When mannose was supplied to leaf discs to sequester cytoplasmic inorganic phosphate, promotion of starch synthesis, and inhibition of CO 2 fixation, were observed in control discs, but not in discs from treated plants. Phosphate buffer reduced starch synthesis in the latter, but not the former discs. The findings suggest that sink removal led to a decreased ratio inorganic phosphate:phosphorylated compounds. In beans 14 C in sugar phosphates increased following sink removal, but without sucrose accumulation, suggesting tighter feedback control of sugar level. Starch accumulated to higher levels than in the other plants, but CO 2 fixation rate was constant for several days

  17. Relationships between soluble sugar concentrations in roots and ecosystem stress for first-year sugar maple seedlings

    Energy Technology Data Exchange (ETDEWEB)

    McLaughlin, J.W.; Reed, D.D.; Jurgensen, M.F.; Mroz, G.D.; Bagley, S.T. [Michigan Technological University, Houghton, MI (United States). School of Forestry and Wood Products

    1996-03-01

    Accumulation of reducing sugars (i.e. glucose and fructose) in plant roots has been consistently correlated with forest dieback and decline and, therefore, has potential as a biological indicator of ecosystem stress. In this study, the relationships between acidic deposition and `natural` (temperature, mycorrhizae, and nutrition) factors with first-year sugar maple seedling root sugar concentrations and growth were assessed in two sugar maple dominated forests in Michigan. Seedlings at the southern site (Wellston) had greater root growth, phosphorus, total sugar, and sucrose concentrations in roots, but lower reducing sugar concentration in roots. In addition, percent root length colonized by vesicular-arbuscular mycorrhizal fungi was less than that found for seedlings growing at the northern site (Alberta). Throughfall deposition of nitrate, sulfate, and hydrogen ions was not significantly correlated with seedling total or reducing sugar concentration. Total sugar concentration in seedling roots was positively correlated with air and soil temperatures at the southern site, but not at the northern site. Seedling tissue phosphorus concentration was correlated with total sugars at both sites, with sucrose at the southern site, and reducing sugars at the northern site. Mycorrhizal colonization rates at the Alberta site were positively correlated with reducing sugar concentration in seedling roots and negatively correlated with sucrose concentration. The results suggest that differences in seedling root sugar concentrations in these two forests are related to seedling root growth and are most likely due to ecological variables, such as available soil phosphorus, temperature, and growing season length through some complex interaction with mycorrhizae rather than acidic deposition stress. 56 refs., 3 figs.

  18. Improved sugar yields from biomass sorghum feedstocks: comparing low-lignin mutants and pretreatment chemistries.

    Science.gov (United States)

    Godin, Bruno; Nagle, Nick; Sattler, Scott; Agneessens, Richard; Delcarte, Jérôme; Wolfrum, Edward

    2016-01-01

    For biofuel production processes to be economically efficient, it is essential to maximize the production of monomeric carbohydrates from the structural carbohydrates of feedstocks. One strategy for maximizing carbohydrate production is to identify less recalcitrant feedstock cultivars by performing some type of experimental screening on a large and diverse set of candidate materials, or by identifying genetic modifications (random or directed mutations or transgenic plants) that provide decreased recalcitrance. Economic efficiency can also be increased using additional pretreatment processes such as deacetylation, which uses dilute NaOH to remove the acetyl groups of hemicellulose prior to dilute acid pretreatment. In this work, we used a laboratory-scale screening tool that mimics relevant thermochemical pretreatment conditions to compare the total sugar yield of three near-isogenic brown midrib ( bmr ) mutant lines and the wild-type (WT) sorghum cultivar. We then compared results obtained from the laboratory-scale screening pretreatment assay to a large-scale pretreatment system. After pretreatment and enzymatic hydrolysis, the bmr mutants had higher total sugar yields than the WT sorghum cultivar. Increased pretreatment temperatures increased reactivity for all sorghum samples reducing the differences observed at lower reaction temperatures. Deacetylation prior to dilute acid pretreatment increased the total sugar yield for all four sorghum samples, and reduced the differences in total sugar yields among them, but solubilized a sizable fraction of the non-structural carbohydrates. The general trends of increased total sugar yield in the bmr mutant compared to the WT seen at the laboratory scale were observed at the large-scale system. However, in the larger reactor system, the measured total sugar yields were lower and the difference in total sugar yield between the WT and bmr sorghum was larger. Sorghum bmr mutants, which have a reduced lignin content showed

  19. Sphingosine-1-Phosphate Lyase Deficient Cells as a Tool to Study Protein Lipid Interactions.

    Directory of Open Access Journals (Sweden)

    Mathias J Gerl

    Full Text Available Cell membranes contain hundreds to thousands of individual lipid species that are of structural importance but also specifically interact with proteins. Due to their highly controlled synthesis and role in signaling events sphingolipids are an intensely studied class of lipids. In order to investigate their metabolism and to study proteins interacting with sphingolipids, metabolic labeling based on photoactivatable sphingoid bases is the most straightforward approach. In order to monitor protein-lipid-crosslink products, sphingosine derivatives containing a reporter moiety, such as a radiolabel or a clickable group, are used. In normal cells, degradation of sphingoid bases via action of the checkpoint enzyme sphingosine-1-phosphate lyase occurs at position C2-C3 of the sphingoid base and channels the resulting hexadecenal into the glycerolipid biosynthesis pathway. In case the functionalized sphingosine looses the reporter moiety during its degradation, specificity towards sphingolipid labeling is maintained. In case degradation of a sphingosine derivative does not remove either the photoactivatable or reporter group from the resulting hexadecenal, specificity towards sphingolipid labeling can be achieved by blocking sphingosine-1-phosphate lyase activity and thus preventing sphingosine derivatives to be channeled into the sphingolipid-to-glycerolipid metabolic pathway. Here we report an approach using clustered, regularly interspaced, short palindromic repeats (CRISPR-associated nuclease Cas9 to create a sphingosine-1-phosphate lyase (SGPL1 HeLa knockout cell line to disrupt the sphingolipid-to-glycerolipid metabolic pathway. We found that the lipid and protein compositions as well as sphingolipid metabolism of SGPL1 knock-out HeLa cells only show little adaptations, which validates these cells as model systems to study transient protein-sphingolipid interactions.

  20. Life Cycle Assessment of Sugar Production (VB)

    DEFF Research Database (Denmark)

    Teljigovic, Mehmed; Mengiardi, Jon; Factor, Gabriela

    1999-01-01

    The environmental organisation NOAH has proposed carrying out an environmental assessment of two different sugar productions (using sugar beet or sugar cane) in order to illustrate which of the systems has a higher environmental impact for sugar consumption in Denmark. Therefore a comparison...... will be made between sugar from sugar beet produced in Denmark versus sugar produces from sugar cane in a tropical country, Brazil, and transported afterwards to Denmark. To evaluate the environmental aspects of these two product systems a Life Cycle Assessement (LCA) will be carried out.From the results...... obtained in the present LCA of sugar produces from sugar canes or sugar beet it is difficult to make an immediate choice between the two possibilities. Indeed, Quantitative results from the EDIP (Environmental Design of Industrial Products) software are globally similar for both ways of producing sugar...

  1. Metal complex derivatives of hydrogen uranyl phosphate

    International Nuclear Information System (INIS)

    Grohol, D.; Blinn, E.L.

    1994-01-01

    Derivatives of hydrogen uranyl phosphate were prepared by incorporating transition metal complexes into the uranyl phosphate matrix. The transition metal complexes employed include bis(ethylenediamine)copper(II), bis(1,3-propanediamine)copper(II) chloride, (triethylenetetramine)copper(II), (1,4,8,11-tetraazacyclotetradecane)copper(II), (1,4,8,12-tetraazacyclopentadecane)copper(II), (1,4,8,11-tetraazacyclotetradecane)nickel(II) chloride, (triethylenetetramine)nickel(II) and others. The chemical analyses of these derivatives indicated that the incorporation of the transition metal complexes into the uranyl phosphate matrix via ion exchange was not stoichiometric. The extent of ion exchange is dependent on the size and structure of the transition metal complex. All complexes were characterized by X-ray powder diffractometry, electronic and infrared spectra, thermal analyses and chemical analysis. An attempt was made to correlate the degree of quenching of the luminescence of the uranyl ion to the spacing between the uranyl phosphate layers in the derivatives

  2. Evaluation of the fermentation of high gravity thick sugar beet juice worts for efficient bioethanol production

    Science.gov (United States)

    2013-01-01

    Background Sugar beet and intermediates of sugar beet processing are considered to be very attractive feedstock for ethanol production due to their content of fermentable sugars. In particular, the processing of the intermediates into ethanol is considerably facilitated because it does not require pretreatment or enzymatic treatment in contrast to production from starch raw materials. Moreover, the advantage of thick juice is high solid substance and saccharose content which eliminates problems with the storability of this feedstock. Results The objective of this study were to investigate bioethanol production from thick juice worts and the effects of their concentration, the type of mineral supplement, as well as the dose of yeast inoculum on fermentation dynamics and ethanol yield. The obtained results show that to ensure efficient ethanolic fermentation of high gravity thick juice worts, one needs to use a yeast strain with high ethanol tolerance and a large amount of inoculum. The highest ethanol yield (94.9 ± 2.8% of the theoretical yield) and sugars intake of 96.5 ± 2.9% were obtained after the fermentation of wort with an extract content of 250 g/kg supplemented with diammonium hydrogen phosphate (0.3 g/L of wort) and inoculated with 2 g of Ethanol Red dry yeast per L of wort. An increase in extract content in the fermentation medium from 250 g/L to 280 g/kg resulted in decreased efficiency of the process. Also the distillates originating from worts with an extract content of 250 g/kg were characterized by lower acetaldehyde concentration than those obtained from worts with an extract content of 280 g/kg. Conclusions Under the favorable conditions determined in our experiments, 38.9 ± 1.2 L of 100% (v/v) ethyl alcohol can be produced from 100 kg of thick juice. The obtained results show that the selection of process conditions and the yeast for the fermentation of worts with a higher sugar content can improve the economic performance of the

  3. Application of Potential Phosphate-Solubilizing Bacteria and Organic Acids on Phosphate Solubilization from Phosphate Rock in Aerobic Rice

    Directory of Open Access Journals (Sweden)

    Qurban Ali Panhwar

    2013-01-01

    Full Text Available A study was conducted at Universiti Putra Malaysia to determine the effect of phosphate-solubilizing bacteria (PSB and organic acids (oxalic & malic on phosphate (P solubilization from phosphate rock (PR and growth of aerobic rice. Four rates of each organic acid (0, 10, 20, and 30 mM, and PSB strain (Bacillus sp. were applied to aerobic rice. Total bacterial populations, amount of P solubilization, P uptake, soil pH, and root morphology were determined. The results of the study showed significantly high P solubilization in PSB with organic acid treatments. Among the two organic acids, oxalic acid was found more effective compared to malic acid. Application of oxalic acid at 20 mM along with PSB16 significantly increased soluble soil P (28.39 mg kg−1, plant P uptake (0.78 P pot−1, and plant biomass (33.26 mg. Addition of organic acids with PSB and PR had no influence on soil pH during the planting period. A higher bacterial population was found in rhizosphere (8.78 log10 cfu g−1 compared to the nonrhizosphere and endosphere regions. The application of organic acids along with PSB enhanced soluble P in the soil solution, improved root growth, and increased plant biomass of aerobic rice seedlings without affecting soil pH.

  4. Insights into the "pair of sugar tongs" surface binding site in barley alpha-amylase isozymes and crystallization of appropriate sugar tongs mutants

    DEFF Research Database (Denmark)

    Tranier, S.; Deville, K.; Robert, X.

    2005-01-01

    of an additional surface binding site called a "pair of sugar tongs" due to the sugar capturing by Tyr380 which is situated in domain C of AMYL For the first time, a biological role for the domain C was suggested as well as a hypothetical explanation of enzymatic differences between the two barley a......-amylase isozymes. However, no sugar was bound at the "sugar tongs" site in the AMY2/acarbose complex. Comparative studies of this domain on the basis of sequence, secondary structure and spatial organization allow to propose factors needed for such a site. One of the most obvious is the replacement of Ser378(AMY1......, surface plasmon resonance sugar binding experiments have proven unambiguously that this residue cannot totally explain the lack of the "pair of sugar tongs" and other tracks must be studied as, for example, the differences in orientation of Asp381 and the critical role of His395, both good candidates...

  5. Phosphorus release from phosphate rock and iron phosphate by low-molecular-weight organic acids.

    Science.gov (United States)

    Xu, Ren-kou; Zhu, Yong-guan; Chittleborough, David

    2004-01-01

    Low-molecular-weight(LMW) organic acids widely exist in soils, particularly in the rhizosphere. A series of batch experiments were carried out to investigate the phosphorus release from rock phosphate and iron phosphate by low-molecular-weight organic acids. Results showed that citric acid had the highest capacity to solubilize P from both rock and iron phosphate. P solubilization from rock phosphate and iron phosphate resulted in net proton consumption. P release from rock phosphate was positively correlated with the pKa values. P release from iron phosphate was positively correlated with Fe-organic acid stability constants except for aromatic acids, but was notcorrelated with pKa. Increase in the concentrations of organic acids enhanced P solubilization from both rock and iron phosphate almost linearly. Addition of phenolic compounds further increased the P release from iron phosphate. Initial solution pH had much more substantial effect on P release from rock phosphate than from iron phosphate.

  6. Sources of Added Sugars in Young Children, Adolescents, and Adults with Low and High Intakes of Added Sugars.

    Science.gov (United States)

    Bailey, Regan L; Fulgoni, Victor L; Cowan, Alexandra E; Gaine, P Courtney

    2018-01-17

    High intake of added sugars is associated with excess energy intake and poorer diet quality. The objective of this cross-sectional study ( n = 16,806) was to estimate usual intakes and the primary food sources of added sugars across the range of intakes (i.e., deciles) among U.S. children (2-8 years), adolescents and teens (9-18 years), and adults (≥19 years) using the National Health and Nutrition Examination (NHANES) data from 2009-2012. The percent energy contributed by added sugars was 14.3 ± 0.2% (2-8 years), 16.2 ± 0.2% (9-18 years), and 13.1 ± 0.2% (≥19 years), suggesting the highest intakes are among adolescents and teens. However, the primary foods/beverages that contribute to added sugars were remarkably consistent across the range of intakes, with the exception of the lowest decile, and include sweetened beverages and sweet bakery products. Interestingly across all age groups, even those in the lowest decile of added sugars exceed the 10% guidelines. Additional foods contributing to high intakes were candy and other desserts (e.g., ice cream) in children and adolescents, and coffee and teas in adults. Tailoring public health messaging to reduce intakes of these identified food groups may be of utility in designing effective strategies to reduce added sugar intake in the U.S.

  7. Sources of Added Sugars in Young Children, Adolescents, and Adults with Low and High Intakes of Added Sugars

    Directory of Open Access Journals (Sweden)

    Regan L. Bailey

    2018-01-01

    Full Text Available High intake of added sugars is associated with excess energy intake and poorer diet quality. The objective of this cross-sectional study (n = 16,806 was to estimate usual intakes and the primary food sources of added sugars across the range of intakes (i.e., deciles among U.S. children (2–8 years, adolescents and teens (9–18 years, and adults (≥19 years using the National Health and Nutrition Examination (NHANES data from 2009–2012. The percent energy contributed by added sugars was 14.3 ± 0.2% (2–8 years, 16.2 ± 0.2% (9–18 years, and 13.1 ± 0.2% (≥19 years, suggesting the highest intakes are among adolescents and teens. However, the primary foods/beverages that contribute to added sugars were remarkably consistent across the range of intakes, with the exception of the lowest decile, and include sweetened beverages and sweet bakery products. Interestingly across all age groups, even those in the lowest decile of added sugars exceed the 10% guidelines. Additional foods contributing to high intakes were candy and other desserts (e.g., ice cream in children and adolescents, and coffee and teas in adults. Tailoring public health messaging to reduce intakes of these identified food groups may be of utility in designing effective strategies to reduce added sugar intake in the U.S.

  8. Sources of Added Sugars in Young Children, Adolescents, and Adults with Low and High Intakes of Added Sugars

    Science.gov (United States)

    Fulgoni, Victor L.; Cowan, Alexandra E.; Gaine, P. Courtney

    2018-01-01

    High intake of added sugars is associated with excess energy intake and poorer diet quality. The objective of this cross-sectional study (n = 16,806) was to estimate usual intakes and the primary food sources of added sugars across the range of intakes (i.e., deciles) among U.S. children (2–8 years), adolescents and teens (9–18 years), and adults (≥19 years) using the National Health and Nutrition Examination (NHANES) data from 2009–2012. The percent energy contributed by added sugars was 14.3 ± 0.2% (2–8 years), 16.2 ± 0.2% (9–18 years), and 13.1 ± 0.2% (≥19 years), suggesting the highest intakes are among adolescents and teens. However, the primary foods/beverages that contribute to added sugars were remarkably consistent across the range of intakes, with the exception of the lowest decile, and include sweetened beverages and sweet bakery products. Interestingly across all age groups, even those in the lowest decile of added sugars exceed the 10% guidelines. Additional foods contributing to high intakes were candy and other desserts (e.g., ice cream) in children and adolescents, and coffee and teas in adults. Tailoring public health messaging to reduce intakes of these identified food groups may be of utility in designing effective strategies to reduce added sugar intake in the U.S. PMID:29342109

  9. Phosphate homeostasis in Bartter syndrome: a case-control study.

    Science.gov (United States)

    Bettinelli, Alberto; Viganò, Cristina; Provero, Maria Cristina; Barretta, Francesco; Albisetti, Alessandra; Tedeschi, Silvana; Scicchitano, Barbara; Bianchetti, Mario G

    2014-11-01

    Bartter patients may be hypercalciuric. Additional abnormalities in the metabolism of calcium, phosphate, and calciotropic hormones have occasionally been reported. The metabolism of calcium, phosphate, and calciotropic hormones was investigated in 15 patients with Bartter syndrome and 15 healthy subjects. Compared to the controls, Bartter patients had significantly reduced plasma phosphate {mean [interquartile range]:1.29 [1.16-1.46] vs. 1.61 [1.54-1.67] mmol/L} and maximal tubular phosphate reabsorption (1.16 [1.00-1.35] vs. 1.41 [1.37-1.47] mmol/L) and significantly increased parathyroid hormone (PTH) level (6.1 [4.5-7.7] vs. 2.8 [2.2-4.4] pmol/L). However, patients and controls did not differ in blood calcium, 25-hydroxyvitamin D, alkaline phosphatase, and osteocalcin levels. In patients, an inverse correlation (P Bartter patients.

  10. A rapid review examining purchasing changes resulting from fiscal measures targeted at high sugar foods and sugar-sweetened drinks.

    Science.gov (United States)

    Roberts, Katharine E; Ells, Louisa J; McGowan, Victoria J; Machaira, Theodora; Targett, Victoria C; Allen, Rachel E; Tedstone, Alison E

    2017-12-15

    To aim of the review was to examine the most recent (2010 onwards) research evidence on the health and behavioural impacts, in adults and children, of fiscal strategies that target high sugar foods and sugar-sweetened drinks (SSDs). A pragmatic rapid review was undertaken using a systematic search strategy. The review was part of a programme of work to support policy development in relation to high sugar food and SSDs. A total of 11 primary research publications were included, describing evidence from France (n = 1), the Netherlands (n = 3), and the United States of America (n = 7), assessed through a variety of study designs, with the majority in adult populations (n = 10). The evidence reviewed focused on consumer behaviour outcomes and suggested that fiscal strategies can influence purchases of high sugar products. Although the majority of studies (n = 10), including three field studies, demonstrated that an increase in the price of high sugar foods and SSDs resulted in a decrease in purchases, eight studies were conducted in a laboratory or virtual setting which may not reflect real-life situations.Findings from this review support evidence from the broader literature that suggests that fiscal measures can be effective in influencing the purchasing of high sugar foods and SSDs.

  11. The Effect of Moderate Dietary Protein and Phosphate Restriction on Calcium-Phosphate Homeostasis in Healthy Older Cats.

    Science.gov (United States)

    Geddes, R F; Biourge, V; Chang, Y; Syme, H M; Elliott, J

    2016-09-01

    Dietary phosphate and protein restriction decreases plasma PTH and FGF-23 concentrations and improves survival time in azotemic cats, but has not been examined in cats that are not azotemic. Feeding a moderately protein- and phosphate-restricted diet decreases PTH and FGF-23 in healthy older cats and thereby slows progression to azotemic CKD. A total of 54 healthy, client-owned cats (≥ 9 years). Prospective double-blinded randomized placebo-controlled trial. Cats were assigned to test diet (protein 76 g/Mcal and phosphate 1.6 g/Mcal) or control diet (protein 86 g/Mcal and phosphate 2.6 g/Mcal) and monitored for 18 months. Changes in variables over time and effect of diet were assessed by linear mixed models. A total of 26 cats ate test diet and 28 cats ate control diet. There was a significant effect of diet on urinary fractional excretion of phosphate (P = 0.045), plasma PTH (P = 0.005), and ionized calcium concentrations (P = 0.018), but not plasma phosphate, FGF-23, or creatinine concentrations. Plasma PTH concentrations did not significantly change in cats fed the test diet (P = 0.62) but increased over time in cats fed the control diet (P = 0.001). There was no significant treatment effect of the test diet on development of azotemic CKD (3 of 26 (12%) test versus 3 of 28 (11%) control, odds ratio 1.09 (95% CI 0.13-8.94), P = 0.92). Feeding a moderately protein- and phosphate-restricted diet has effects on calcium-phosphate homeostasis in healthy older cats and is well tolerated. This might have an impact on renal function and could be useful in early chronic kidney disease. Copyright © 2016 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  12. A trial of sugar-free or sugar-sweetened beverages and body weight in children

    NARCIS (Netherlands)

    de Ruyter, Janne C; Olthof, Margreet R; Seidell, Jacob C; Katan, Martijn B

    2012-01-01

    BACKGROUND: The consumption of beverages that contain sugar is associated with overweight, possibly because liquid sugars do not lead to a sense of satiety, so the consumption of other foods is not reduced. However, data are lacking to show that the replacement of sugar-containing beverages with

  13. Sugar palm ethanol. Analysis of economic feasibility and sustainability

    Energy Technology Data Exchange (ETDEWEB)

    Van de Staaij, J.; Van den Bos, A.; Hamelinck, C. [Ecofys Netherlands, Utrecht (Netherlands); Martini, E.; Roshetko, J.; Walden, D. [Winrock, Little Rock, AR (United States)

    2011-08-15

    This study evaluates whether sugar palm is a suitable crop for biofuels and how production of ethanol from sugar palm in a large-scale setting is sustainable and economically feasible. Key questions are: Are the assumed high yields realistic in practice for sustained periods in largescale plantations?; Can sugar palm indeed compete economically with other crops for biofuels?; What are the effects of large-scale cultivation and processing of sugar palm for the natural environment and the local community? To answer these questions, Ecofys and Winrock have assessed the feasibility of largescale sugar palm cultivation for the production of ethanol using empirical data from existing sugar palm plantings. We analysed two production models to investigate the range of outcomes when varying important parameters: (1) a conservative system, whereby sugar palms are mixed with other crops and (2) an intensive system to explore the theoretical maximum yield when solely focusing on sugar palm. As background, Chapter 2 first describes the process of sugar palm cultivation, the 'tapping' and conversion into ethanol. Chapter 3 describes the data collection by Winrock. It presents an overview of the collected field data and explains the main empirical findings. Chapter 4 elaborates the two production systems and presents the results of the economic analyses (summarized in cash flow diagrams showing the timing of costs and benefits). Chapter 5 analyses the possible sustainability risks and benefits of sugar palm ethanol and investigates the integration possibilities of sugar palm in agro-forestry systems with other crops. Finally, Chapter 6 concludes by evaluating the potential of sugar palm as a source of biofuel and providing recommendations.

  14. [Assays of HbA1c and Amadori products in human biology].

    Science.gov (United States)

    Gillery, P

    2014-09-01

    Different Amadori products, formed during the early steps of the non-enzymatic glycation of proteins, may be assayed in current practice in human biology. The most important marker is HbA1c, resulting from the binding of glucose to the N-terminal extremity of HbA beta chains. HbA1c may be evaluated by various techniques (ion exchange or affinity high performance liquid chromatography, capillary electrophoresis, immunoassay, enzymatic technique) and is considered the best marker of diabetic patient survey. Due to its irreversible and cumulative formation, it provides a retrospective information on the glycemic balance over the four to eight weeks preceding blood collection. It benefits from an international standardization, based on a reference method using liquid chromatography coupled to capillary electrophoresis or mass spectrometry, maintained by an international network of reference laboratories. When HbA1c assay cannot be used (anemia, hemolysis, hemoglobinopathy) or when a shorter period of glycemic equilibrium must be evaluated (child and adolescent, pregnancy, therapeutic changes), other Amadori products may be assayed, like plasma fructosamine (all plasma glycated proteins) or glycated albumin. Nevertheless, these assays are less used in practice, because their semiological value has been less evidenced. Besides, fructosamine assay lacks specificity, and glycated albumin assay has been described recently. An expanding use of HbA1c assay is expected, especially for the diagnosis of diabetes mellitus and the evaluation of other risks, especially cardiovascular ones. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  15. Development and Fit-for-Purpose Validation of a Soluble Human Programmed Death-1 Protein Assay.

    Science.gov (United States)

    Ni, Yan G; Yuan, Xiling; Newitt, John A; Peterson, Jon E; Gleason, Carol R; Haulenbeek, Jonathan; Santockyte, Rasa; Lafont, Virginie; Marsilio, Frank; Neely, Robert J; DeSilva, Binodh; Piccoli, Steven P

    2015-07-01

    Programmed death-1 (PD-1) protein is a co-inhibitory receptor which negatively regulates immune cell activation and permits tumors to evade normal immune defense. Anti-PD-1 antibodies have been shown to restore immune cell activation and effector function-an exciting breakthrough in cancer immunotherapy. Recent reports have documented a soluble form of PD-1 (sPD-1) in the circulation of normal and disease state individuals. A clinical assay to quantify sPD-1 would contribute to the understanding of sPD-1-function and facilitate the development of anti-PD-1 drugs. Here, we report the development and validation of a sPD-1 protein assay. The assay validation followed the framework for full validation of a biotherapeutic pharmacokinetic assay. A purified recombinant human PD-1 protein was characterized extensively and was identified as the assay reference material which mimics the endogenous analyte in structure and function. The lower limit of quantitation (LLOQ) was determined to be 100 pg/mL, with a dynamic range spanning three logs to 10,000 pg/mL. The intra- and inter-assay imprecision were ≤15%, and the assay bias (percent deviation) was ≤10%. Potential matrix effects were investigated in sera from both normal healthy volunteers and selected cancer patients. Bulk-prepared frozen standards and pre-coated Streptavidin plates were used in the assay to ensure consistency in assay performance over time. This assay appears to specifically measure total sPD-1 protein since the human anti-PD-1 antibody, nivolumab, and the endogenous ligands of PD-1 protein, PDL-1 and PDL-2, do not interfere with the assay.

  16. SNPs in genes functional in starch-sugar interconversion associate with natural variation of tuber starch and sugar content of potato (Solanum tuberosum L.).

    Science.gov (United States)

    Schreiber, Lena; Nader-Nieto, Anna Camila; Schönhals, Elske Maria; Walkemeier, Birgit; Gebhardt, Christiane

    2014-07-31

    Starch accumulation and breakdown are vital processes in plant storage organs such as seeds, roots, and tubers. In tubers of potato (Solanum tuberosum L.) a small fraction of starch is converted into the reducing sugars glucose and fructose. Reducing sugars accumulate in response to cold temperatures. Even small quantities of reducing sugars affect negatively the quality of processed products such as chips and French fries. Tuber starch and sugar content are inversely correlated complex traits that are controlled by multiple genetic and environmental factors. Based on in silico annotation of the potato genome sequence, 123 loci are involved in starch-sugar interconversion, approximately half of which have been previously cloned and characterized. By means of candidate gene association mapping, we identified single-nucleotide polymorphisms (SNPs) in eight genes known to have key functions in starch-sugar interconversion, which were diagnostic for increased tuber starch and/or decreased sugar content and vice versa. Most positive or negative effects of SNPs on tuber-reducing sugar content were reproducible in two different collections of potato cultivars. The diagnostic SNP markers are useful for breeding applications. An allele of the plastidic starch phosphorylase PHO1a associated with increased tuber starch content was cloned as full-length cDNA and characterized. The PHO1a-HA allele has several amino acid changes, one of which is unique among all known starch/glycogen phosphorylases. This mutation might cause reduced enzyme activity due to impaired formation of the active dimers, thereby limiting starch breakdown. Copyright © 2014 Schreiber et al.

  17. Emerging Role of Sphingosine-1-phosphate in Inflammation, Cancer, and Lymphangiogenesis

    Directory of Open Access Journals (Sweden)

    Kazuaki Takabe

    2013-07-01

    Full Text Available The main function of the lymphatic system is to control and maintain fluid homeostasis, lipid transport, and immune cell trafficking. In recent years, the pathological roles of lymphangiogenesis, the generation of new lymphatic vessels from preexisting ones, in inflammatory diseases and cancer progression are beginning to be elucidated. Sphingosine-1-phosphate (S1P, a bioactive lipid, mediates multiple cellular events, such as cell proliferation, differentiation, and trafficking, and is now known as an important mediator of inflammation and cancer. In this review, we will discuss recent findings showing the emerging role of S1P in lymphangiogenesis, in inflammation, and in cancer.

  18. Thirteen years of observations on primary sugars and sugar alcohols over remote Chichijima Island in the western North Pacific

    Directory of Open Access Journals (Sweden)

    S. K. Verma

    2018-01-01

    Full Text Available In order to understand the atmospheric transport of bioaerosols, we conducted long-term observations of primary sugars and sugar alcohols over remote Chichijima Island in the western North Pacific from 2001 to 2013. Our results showed that concentrations of total sugar compounds for 13 years ranged from 1.2 to 310 ng m−3 (average of 46 ± 49 ng m−3. We found that atmospheric circulations significantly affect the seasonal variations of bioaerosol distributions over the western North Pacific. The primary sugars (glucose and fructose maximized in summer, possibly due to an increased emission of the vegetation products from local vascular plants in Chichijima. We also found higher concentrations of sugar components (arabitol, mannitol, and trehalose in more recent years during summer and autumn, suggesting an enhanced emission of fungal and microbial species over the island. Sucrose peaked in late winter to early spring, indicating a springtime pollen contribution by long-range atmospheric transport, while elevated concentrations of sucrose in early summer could be explained by long-range transport of soil dust from Southeast Asia to Chichijima. Sucrose and trehalose were found to present increasing trends from 2001 to 2013, while total sugar components did not show any clear trends during the 13-year period. Positive matrix factorization analyses suggested the locally emitted sugar compounds as well as long-range-transported airborne pollen grains, microbes, and fungal spores are the major contributors to total sugar compounds in the Chichijima aerosols. Backward air mass trajectories support the atmospheric transport of continental aerosols from the Asian continent during winter and spring over Chichijima.

  19. The government policy related to sugar-sweetened beverages in Indonesia

    OpenAIRE

    Mohamad Thahir Haning; Andi Imam Arundhana; Asry Dwi Muqni

    2016-01-01

    Background: There are several options to enforce reduction in the use of sugary drinks such as strengthening regulations, taxation on the products and food labeling.  Aims & Objectives: 1) Identify the policy in Indonesia that regulates the quantity and the use of sugar in a beverage product; 2) Describe the sugar content in sugar-sweetened beverages (SSB) and its impact on human health. Material & Methods: Literature search on sugar use and tax policies on SSB was conducted and 6 rel...

  20. CDE-1 affects chromosome segregation through uridylation of CSR-1-bound siRNAs.

    Science.gov (United States)

    van Wolfswinkel, Josien C; Claycomb, Julie M; Batista, Pedro J; Mello, Craig C; Berezikov, Eugene; Ketting, René F

    2009-10-02

    We have studied the function of a conserved germline-specific nucleotidyltransferase protein, CDE-1, in RNAi and chromosome segregation in C. elegans. CDE-1 localizes specifically to mitotic chromosomes in embryos. This localization requires the RdRP EGO-1, which physically interacts with CDE-1, and the Argonaute protein CSR-1. We found that CDE-1 is required for the uridylation of CSR-1 bound siRNAs, and that in the absence of CDE-1 these siRNAs accumulate to inappropriate levels, accompanied by defects in both meiotic and mitotic chromosome segregation. Elevated siRNA levels are associated with erroneous gene silencing, most likely through the inappropriate loading of CSR-1 siRNAs into other Argonaute proteins. We propose a model in which CDE-1 restricts specific EGO-1-generated siRNAs to the CSR-1 mediated, chromosome associated RNAi pathway, thus separating it from other endogenous RNAi pathways. The conserved nature of CDE-1 suggests that similar sorting mechanisms may operate in other animals, including mammals.

  1. Stability study for magnetic reagent assaying Hb and HbA1c

    Energy Technology Data Exchange (ETDEWEB)

    Hsieh, Wen-Pin [Actherm Inc., Hsinchu 200, Taiwan (China); Chieh, J.J.; Yang, C.C. [Institute of Electro-optical Science and Technology, National Taiwan Normal University, Taipei 116, Taiwan (China); Yang, S.Y. [Institute of Electro-optical Science and Technology, National Taiwan Normal University, Taipei 116, Taiwan (China); MagQu Co., Ltd., Sindian Dist., New Taipei City 231, Taiwan (China); Chen, Po-Yu; Huang, Yu-Hao [Actherm Inc., Hsinchu 200, Taiwan (China); Hong, Y.W. [Institute of Electro-optical Science and Technology, National Taiwan Normal University, Taipei 116, Taiwan (China); Horng, H.E., E-mail: phyfv001@ntnu.edu.tw [Institute of Electro-optical Science and Technology, National Taiwan Normal University, Taipei 116, Taiwan (China)

    2013-01-15

    Reagents for magnetically labeled immunoassay on human Hb and human HbA1c have been synthesized. The reagents consist of Fe{sub 3}O{sub 4} magnetic particles biofunctionalized with antibodies against Hb and HbA1c. It has been demonstrated that the reagents can be applied to quantitatively detect Hb and HbA1c by using immunomagnetic reduction assay. In addition to characterizing the assay properties, such as the standard curve and the low-detection limit, the stability of reagents is investigated. To do this, the temporal dependence of particle sizes and the bio-activity of reagents are monitored. The results show that the reagents are highly stable when stored at 2-8 Degree-Sign C. This means that the reagents synthesized in this work are promising for practical applications. - Highlights: Black-Right-Pointing-Pointer The properties of assaying Hb and HbA1c using immunomagnetic reduction are studied. Black-Right-Pointing-Pointer The magnetic nanoparticles with antibodies are highly stable in solutions. Black-Right-Pointing-Pointer No significant mutual interference between Hb and HbA1c in assays is observed. Black-Right-Pointing-Pointer High-sensitivity assays on Hb and HbA1c using immunomagnetic reduction are achieved.

  2. Effects of Balneotherapy on Serum Levels of Shingosine-1-Phosphate in Patients With Osteoarthritis.

    Science.gov (United States)

    Ustyol, Esra Aycan; Karaarslan, Fatih; Bekpinar, Seldag; Ozkuk, Kagan; Erdogan, Nergis

    2017-11-01

    Context • Balneotherapy is one of the most commonly used nonpharmacological interventions for osteoarthritis (OA), but its mechanism of action in relieving pain and stiffness and in improving physical function is not well understood. Studies have found that therapy provokes a series of neuroendocrinal reactions with anti-inflammatory and analgesic effects. Sphingosine-1-phosphate (S1P), a bioactive lipid, has been implicated as an important mediator in the maintenance of physiological processes (eg, vascular barrier integrity) and in pathophysiologic processes such as inflammatory conditions. Accordingly, targeting S1P and S1P receptors may offer a potential therapy for arthritis. Objective • The aims of the present study were to determine whether (1) balneotherapy modified the circulating levels of S1P as well as some inflammatory parameters and stress markers, in patients with OA; and (2) to assess the relationship of those parameters to therapeutic efficacy. Design • This study was designed as an uncontrolled longitudinal study. Setting • The study took place at the Bolu Physical Therapy and Rehabilitation Hospital (Bolu, Turkey). Participants • Forty patients who suffered from general OA in at least 3 positions on the body, one of which could be the vertebral column, and who fulfilled the American College of Rheumatology Classification criteria and the Kellgren-Moore radiologic criteria, were enrolled in the intervention group in the study. Intervention • During balneotherapy, the participants were fully immersed in warm thermo-mineral water for 20 min at a temperature of 38°C to 40°C. A total of 15 immersions were performed in a period of 15 d. Outcome Measures • A baseline clinical evaluation of participants' pain, stiffness, and physical function was carried out using the Western Ontario and McMaster Universities questionnaire. Baseline serum levels of S1P, cyclooxygenase 2 (COX-2), matrix metalloproteinase 3 (MMP-3), and heat shock protein

  3. Cerebrospinal Fluid Phosphate in Delirium after Hip Fracture

    Directory of Open Access Journals (Sweden)

    Ane-Victoria Idland

    2017-09-01

    Full Text Available Aims: Phosphate is essential for neuronal activity. We aimed to investigate whether delirium is associated with altered phosphate concentrations in cerebrospinal fluid (CSF and serum. Methods: Seventy-seven patients with hip fracture were assessed for delirium before and after acute surgery. Prefracture dementia was diagnosed by an expert panel. Phosphate was measured in CSF obtained immediately before spinal anesthesia (n = 77 and in serum (n = 47. CSF from 23 cognitively healthy elderly patients undergoing spinal anesthesia was also analyzed. Results: Hip fracture patients with prevalent delirium had higher CSF phosphate concentrations than those without delirium (median 0.63 vs. 0.55 mmol/L, p = 0.001. In analyses stratified on dementia status, this difference was only significant in patients with dementia. Serum phosphate was ∼1 mmol/L; there was no association between serum phosphate concentration and delirium status. CSF phosphate did not correlate with serum levels. Conclusion: Patients with delirium superimposed on dementia have elevated phosphate levels.

  4. A phosphate-dependent shift in redox state of cerium oxide nanoparticles and its effects on catalytic properties

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Sanjay; Dosani, Talib; Karakoti, Ajay S.; Kumar, Amit; Seal, Sudipta; Self, William

    2011-10-01

    Cerium oxide nanoparticles (CeNPs) have shown promise as catalytic antioxidants in cell culture and animal models as both superoxide dismutase and catalase mimetics. The reactivity of the cerium (Ce) atoms at the surface of its oxide particle is critical to such therapeutic properties, yet little is known about the potential for a protein or small molecule corona to form on these materials in vivo. Moreover Ce atoms in these active sites have the potential to interact with small molecule anions, peptides, or sugars when administered in culture or animal models. Several nanomaterials have been shown to alter or aggregate under these conditions, rendering them less useful for biomedical applications. In this work we have studied the change in catalytic properties of CeNPs when exposed to various biologically relevant conditions in vitro. We have found that CeNPs are resistant to broad changes in pH and also not altered by incubation in cell culture medium. However to our surprise phosphate anions significantly altered the characteristics of these nanomaterials and shifted the catalytic behavior due to the binding of phosphate anions to cerium. Given the abundance of phosphate in biological systems in an inorganic form, it is likely that the action of CeNPs as a catalyst may be strongly influenced by the local concentration of phosphate in the cells and/or tissues in which it has been introduced.

  5. Transcription Factor AREB2 Is Involved in Soluble Sugar Accumulation by Activating Sugar Transporter and Amylase Genes.

    Science.gov (United States)

    Ma, Qi-Jun; Sun, Mei-Hong; Lu, Jing; Liu, Ya-Jing; Hu, Da-Gang; Hao, Yu-Jin

    2017-08-01

    Sugars play important roles in plant growth and development, crop yield and quality, as well as responses to abiotic stresses. Abscisic acid (ABA) is a multifunctional hormone. However, the exact mechanism by which ABA regulates sugar accumulation is largely unknown in plants. Here, we tested the expression profile of several sugar transporter and amylase genes in response to ABA treatment. MdSUT2 and MdAREB2 were isolated and genetically transformed into apple ( Malus domestica ) to investigate their roles in ABA-induced sugar accumulation. The MdAREB2 transcription factor was found to bind to the promoters of the sugar transporter and amylase genes and activate their expression. Both MdAREB2 and MdSUT2 transgenic plants produced more soluble sugars than controls. Furthermore, MdAREB2 promoted the accumulation of sucrose and soluble sugars in an MdSUT2 -dependent manner. Our results demonstrate that the ABA-responsive transcription factor MdAREB2 directly activates the expression of amylase and sugar transporter genes to promote soluble sugar accumulation, suggesting a mechanism by which ABA regulates sugar accumulation in plants. © 2017 American Society of Plant Biologists. All Rights Reserved.

  6. Preparation of calcium phosphate paste

    International Nuclear Information System (INIS)

    Mohd Reusmaazran Yusof; Norzita Yaacob; Idris Besar; Che Seman Mahmood; Rusnah Mustafa

    2010-01-01

    Calcium phosphate paste were prepared by mixing between calcium sodium potassium phosphate, Ca 2 NaK (PO 4 ) 2 (CSPP) and monocalcium phosphate monohydrate, Ca(H 2 PO 4 ) 2 .H 2 O (MCPM). CSPP were obtained by reaction between calcium hydrogen phosphate (CaHPO 4 ), potassium carbonate (K 2 CO 3 ) and sodium carbonate (Na 2 CO 3 ) in solid state sintering process followed by quenching in air at 1000 degree Celsius. The paste was aging in simulated body fluid (SBF) for 0.5, 1, 3, 6, 12, 24, 48 hrs, 3, 7 and 14 days. The morphological investigation indicated the formation of apatite crystal were first growth after 24 hours. The obvious growth of apatite crystal was shown at 3 days. The obvious growth of apatite crystal was shown in 7 and 14 days indicated the prediction of paste would have rapid reaction with bone after implantation. (author)

  7. Translation and implementation of added sugars consumption recommendations: a conference report from the American Heart Association Added Sugars Conference 2010.

    Science.gov (United States)

    Van Horn, Linda; Johnson, Rachel K; Flickinger, Brent D; Vafiadis, Dorothea K; Yin-Piazza, Shirley

    2010-12-07

    A 2-day forum was convened to (1) discuss ways to translate the 2009 American Heart Association added sugars recommendations into actions in areas such as regulation, food labeling, nutrient content claims, and practical application in the American diet; (2) review surveillance methodology and metrics for tracking and understanding the impact of reducing added sugars in the diet; and (3) initiate the development of a framework for future collaboration to help Americans implement science-based guidance relative to added sugars. More than 100 multinational participants representing scientists from academia and government and stakeholders engaged in food production, development, and processing, food manufacturing and servicing, food and nutrition policy, and nutrition recommendations for the public attended the conference. Presentations included definitions and examples of added sugars, current US and international added sugars perspectives, added sugars in diets of individuals and in the food supply, food technology behind added sugars, added sugars and health, food manufacturer perspectives, added sugars food-labeling considerations, and examples of positive approaches to improve eating behaviors and the food environment. Facilitated breakout sessions were conducted after the plenary sessions to allow participants to contribute their expertise and thoughts. The American Heart Association Added Sugars Conference is the first step in an important process that facilitates collaboration across science, public health, and industry to foster innovation, partnerships, policy, and implementation of new products and services for the benefit of the health and well-being of the American public. Science has advanced in the area of added sugars and health, creating mounting pressure to use better methods for translation and dissemination of the science for consumer education and for food companies to respond by producing foods and beverages with fewer added sugars. The new

  8. First two-reagent vitamin D assay for general clinical chemistry.

    Science.gov (United States)

    Saida, Fakhri B; Padilla-Chee, Mario; Dou, Chao; Yuan, Chong

    2018-05-01

    Vitamin D is a lipid-soluble molecule that plays key physiological roles in the metabolism of calcium, phosphate and magnesium. Recent studies show that deficiency in vitamin D is linked to cardiovascular diseases, autoimmune diseases and cancer. As a result, regular monitoring of 25-OH vitamin D (the main circulating form of vitamin D) is becoming essential. Current 25-OH vitamin D testing methodologies are cumbersome (too many reagents, long incubation times, phase separation) and are not compatible with general clinical chemistry platforms. Here, we report on a novel method to detect 25-OH vitamin D that is fast (results in 10 min or less), simple (two reagents) and compatible with virtually all general clinical chemistry analyzers. An immunoturbidimetric assay for 25-OH vitamin D (the Diazyme EZ Vitamin D Assay) has been developed using nanoparticles and vitamin D-specific antibodies. The performance of the assay kit, which consists of two reagents and five calibrators, was tested on the Beckman AU680 analyzer (AU680). The new assay was precise, sensitive (LOD = 7.2 nmol/L), linear (up to 390.1 nmol/L) and correlated strongly (R 2  > 0.95) with major commercial 25-OH vitamin D assays. Additionally, the assay was found to be the fastest to date, with the first results obtained within 10 min. Throughput on the AU680 was estimated at over 300 tests per hour. The newly developed 25-OH vitamin D assay is fast, precise and accurate. It can be run on most general chemistry analyzers. This assay aims at providing vitamin D-testing capabilities to all clinical chemistry laboratories. Copyright © 2018 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  9. Food sources of free sugars in children's diet and identification of lifestyle patterns associated with free sugars intake: the GRECO (Greek Childhood Obesity) study.

    Science.gov (United States)

    Farajian, Paul; Risvas, Grigoris; Panagiotakos, Demosthenes B; Zampelas, Antonis

    2016-09-01

    Excessive free sugars consumption has a possible role in health issues, diet quality and obesity development. The present cross-sectional study aimed to identify the major food sources of free sugars in Greek children's diet and investigate possible associations of dietary patterns with free sugars intake. Anthropometric measurements and information on dietary and physical activity habits were obtained. Energy and free sugars intake coming from foods were estimated and principal components analysis was applied to identify dietary patterns. The GRECO (Greek Childhood Obesity) study. Nationwide sample of 3089 children (aged 10-12 years). Adopting WHO criteria, 44·2 % of participants were categorized as having free sugars intake above 10 % of total energy intake. Mean contribution of free sugars to energy intake was 11·2 %, and the major food sources of free sugars differed from those of other childhood populations. Free sugars intake was not associated with overweight/obesity. Multiple linear regression analysis revealed that two lifestyle and dietary patterns, characterized by higher consumption of sweets, fast foods, fries, sugared drinks, frequently ordering/eating outside home and having meals in front of a screen (pattern 1) and higher consumption of whole fruits, 100 % fruit juices, vegetables, legumes and honey/jam (pattern 2), were positively associated with free sugars intake. A large proportion of children exceeded the recommended cut-off and free sugars intake was associated with lifestyle patterns rather than single foods. Public health programmes aiming to reduce free sugars consumption should be tailored on promoting the correct dietary habits of specific childhood populations.

  10. Sphingosine-1-Phosphate Evokes Unique Segment-Specific Vasoconstriction of the Renal Microvasculature

    Science.gov (United States)

    Singletary, Sean T.; Cook, Anthony K.; Hobbs, Janet L.; Pollock, Jennifer S.; Inscho, Edward W.

    2014-01-01

    Sphingosine-1-phosphate (S1P), a bioactive sphingolipid metabolite, has been implicated in regulating vascular tone and participating in chronic and acute kidney injury. However, little is known about the role of S1P in the renal microcirculation. Here, we directly assessed the vasoresponsiveness of preglomerular and postglomerular microvascular segments to exogenous S1P using the in vitro blood-perfused juxtamedullary nephron preparation. Superfusion of S1P (0.001–10 μM) evoked concentration-dependent vasoconstriction in preglomerular microvessels, predominantly afferent arterioles. After administration of 10 μM S1P, the diameter of afferent arterioles decreased to 35%±5% of the control diameter, whereas the diameters of interlobular and arcuate arteries declined to 50%±12% and 68%±6% of the control diameter, respectively. Notably, efferent arterioles did not respond to S1P. The S1P receptor agonists FTY720 and FTY720-phosphate and the specific S1P1 receptor agonist SEW2871 each evoked modest afferent arteriolar vasoconstriction. Conversely, S1P2 receptor inhibition with JTE-013 significantly attenuated S1P-mediated afferent arteriolar vasoconstriction. Moreover, blockade of L-type voltage-dependent calcium channels with diltiazem or nifedipine attenuated S1P-mediated vasoconstriction. Intravenous injection of S1P in anesthetized rats reduced renal blood flow dose dependently. Western blotting and immunofluorescence revealed S1P1 and S1P2 receptor expression in isolated preglomerular microvessels and microvascular smooth muscle cells. These data demonstrate that S1P evokes segmentally distinct preglomerular vasoconstriction via activation of S1P1 and/or S1P2 receptors, partially via L-type voltage-dependent calcium channels. Accordingly, S1P may have a novel function in regulating afferent arteriolar resistance under physiologic conditions. PMID:24578134

  11. Declining consumption of added sugars and sugar-sweetened beverages in Australia: a challenge for obesity prevention.

    Science.gov (United States)

    Brand-Miller, Jennie C; Barclay, Alan W

    2017-04-01

    Background: Reduced intakes of added sugars and sugar-sweetened beverages (SSBs) have been the main focus of efforts to stall obesity. Although obesity has risen steeply in Australia, some evidence suggests that added-sugars and SSB intakes have declined over the same time frame. Objective: We investigated recent trends in the availability of sugars and sweeteners and changes in intakes of total sugars, added sugars, and SSBs in Australia by using multiple, independent data sources. Design: The study was designed to compare relevant data published by the Food and Agriculture Organization of the United Nations [FAO Statistics Division Database (FAOSTAT)], the Australian government, academia, and the food industry. Results: With the use of the FAOSTAT food balance sheets for Australia, the per capita availability of added or refined sugars and sweeteners was shown to have fallen 16% from 152 g/d in 1980 to 127 g/d in 2011 ( P -trend = 0.001). In national dietary surveys in 1995 and 2011-2012, added-sugars intake declined markedly in adult men (from 72 to 59 g/d; -18%) but not in women (44-42 g/d; NS). As a proportion of total energy, added-sugars intake fell 10% in adult men but nonsignificantly in adult women. Between 1995 and 2011-2012, the proportion of energy from SSBs (including 100% juice) declined 10% in adult men and 20% in women. More marked changes were observed in children aged 2-18 y. Data from national grocery sales indicated that per capita added-sugars intakes derived from carbonated soft drinks fell 26% between 1997 and 2011 (from 23 to 17 g/d) with similar trends for noncarbonated beverages. Conclusions: In Australia, 4 independent data sets confirmed shorter- and longer-term declines in the availability and intake of added sugars, including those contributed by SSBs. The findings challenge the widespread belief that energy from added sugars or sugars in solution are uniquely linked to the prevalence of obesity. © 2017 American Society for Nutrition.

  12. Carbohydrates, Sugar, and Your Child

    Science.gov (United States)

    ... carbs are: simple carbohydrates (or simple sugars): including fructose, glucose, and lactose, which also are found in nutritious ... sugar, check the ingredients list for sugar, corn syrup, or other sweeteners, such as dextrose, fructose, honey, or molasses, to name just a few. ...

  13. Proteomic and biochemical basis for enhanced growth yield of Enterobacter sp. LCR1 on insoluble phosphate medium.

    Science.gov (United States)

    Kumar, Arvind; Rai, Lal Chand

    2015-01-01

    Proteomics and biochemical analyses were used to unravel the basis for higher growth yield of Enterobacter sp. LCR1 on insoluble phosphate medium compared to soluble. Proteomic analysis using 2-DE, MALDI-TOF/MS and LC-MS revealed the involvement of nine proteins. Down-regulation of fructose bisphosphate aldolase with decreased concentrations of glucose-6-phosphate and fructose-6-phosphate indicated diminished glycolysis. However, up-regulation of phosphoglycerate mutase, increase in the activities of 6-phosphogluconate dehydratase, 2-keto-3-deoxy-6-phosphogluconate aldolase and 6-phosphogluconate dehydrogenase suggested induction of Entner-Doudoroff and pentose phosphate pathways. These pathways generate sufficient energy from gluconic acid, which is also used for biosynthesis as indicated by up-regulation of elongation factor Tu, elongation factor G and protein disulfide isomerase. Increased reactive oxygen species (ROS) formation resulting from organic acid oxidation leads to overexpressed manganese superoxide dismutase and increased activities of catalase and ascorbate peroxidase. Thus the organism uses gluconate instead of glucose for energy, while alleviating extra ROS formation by oxidative defense enzymes. Copyright © 2014 Elsevier GmbH. All rights reserved.

  14. Potentiality of Acidithiobacillus thiooxidans in Microbial Solubilization of Phosphate Mine Tailings

    Directory of Open Access Journals (Sweden)

    S Dhakar

    2015-04-01

    Full Text Available This paper deals with the solubilization behavior of the tailings produced by the floatation of a complex low grade phosphate ore. The composition of the tailings was essentially dolomite (52.04% with minor amounts of phosphate, iron and aluminium oxides (10.4 and 0.5% respectively. The presence of these products created uncontrolled land pollution and severely affected groundwater. An initiative has been taken up for utilization of this waste to generate an eco-friendly product. First step towards this panorama is incorporation of suitable microorganisms for the biodegradation of this effluent. Sulphur oxidizing bacteria Acidithiobacillus thiooxidans produces sulphuric acid which neutralizes the dolomitic tailings and convert it into plant available forms. The solubilization activity was tested in sulphur medium with 5, 10, 15 and 20% concentration of tailings. The solubilization is graded on the basis of pH, Electrical conductivity (EC, soluble calcium and magnesium and soluble phosphate. The results from ex-situ experiments showed that the treatment with 15% tailings ended with highest solubilization. The values of pH, EC, soluble calcium and magnesium and soluble phosphate for this treatment were 4.92, 31.6 dS/m, 10.8 mL EDTA and 17.24 µg/mL respectively. Also, the results proved that sulphur oxidizing bacteria Acidithiobacillus thiooxidans is capable of solubilizing dolomitic tailings from the Jhamarkotra mines. Finally, an important factor taken into account was solubilization of residual phosphate along with dolomite in the tailings. This combined action affects the solubilization behaviour of the residue, which was also showed successfully with the assayed laboratory studies.

  15. 75 FR 22095 - USDA Reassigns Domestic Cane Sugar Allotments and Increases the Fiscal Year 2010 Raw Sugar Tariff...

    Science.gov (United States)

    2010-04-27

    ... USDA Reassigns Domestic Cane Sugar Allotments and Increases the Fiscal Year 2010 Raw Sugar Tariff-Rate... announced a reassignment of surplus sugar under domestic cane sugar allotments of 200,000 short tons raw value (STRV) to imports, and increased the fiscal year (FY) 2010 raw sugar tariff-rate quota (TRQ) by...

  16. 75 FR 38764 - USDA Reassigns Domestic Cane Sugar Allotments and Increases the Fiscal Year 2010 Raw Sugar Tariff...

    Science.gov (United States)

    2010-07-06

    ... USDA Reassigns Domestic Cane Sugar Allotments and Increases the Fiscal Year 2010 Raw Sugar Tariff-Rate... announced a reassignment of surplus sugar under domestic cane sugar allotments of 300,000 short tons raw value (STRV) to imports, and increased the fiscal year (FY) 2010 raw sugar tariff-rate quota (TRQ) by...

  17. 76 FR 20305 - USDA Reassigns Domestic Cane Sugar Allotments and Increases the Fiscal Year 2011 Raw Sugar Tariff...

    Science.gov (United States)

    2011-04-12

    ... USDA Reassigns Domestic Cane Sugar Allotments and Increases the Fiscal Year 2011 Raw Sugar Tariff-Rate... announced a reassignment of surplus sugar under domestic cane sugar allotments of 325,000 short tons raw value (STRV) to imports, and increased the fiscal year (FY) 2011 raw sugar tariff-rate quota (TRQ) by...

  18. Comparative anatomy of the peduncles of Thai Sugar Palms provides insight on putative sugar transport mechanisms

    DEFF Research Database (Denmark)

    Somjaiai, Pananun; Barfod, Anders; Jampeetong, Arunothai

    Inflorescences of sugar palms exude copious amounts of sugar-rich sap, when their peduncles are scarred. In Thailand this phenomenon form the basis of a widespread cottage industry based on species such as Arenga pinnata, Borassus flabellifera, Cocos nucifera and Nypa fruticans. The extracted sugar...... sap is used mainly for jaggery, syrup and different types of beverages. In this study we looked for anatomical correlates of the elevated sap flow in injured peduncles of sugar palms. Despite a limited sample size we observed that sugar producing palms differ from the reference palm Chamaedorea...

  19. Nano-sized calcium phosphate particles for periodontal gene therapy.

    Science.gov (United States)

    Elangovan, Satheesh; Jain, Shardool; Tsai, Pei-Chin; Margolis, Henry C; Amiji, Mansoor

    2013-01-01

    Growth factors such as platelet-derived growth factor (PDGF) have significantly enhanced periodontal therapy outcomes with a high degree of variability, mostly due to the lack of continual supply for a required period of time. One method to overcome this barrier is gene therapy. The aim of this in vitro study is to evaluate PDGF-B gene delivery in fibroblasts using nano-sized calcium phosphate particles (NCaPP) as vectors. NCaPP incorporating green fluorescent protein (NCaPP-GFP) and PDGF-B (NCaPP-PDGF-B) plasmids were synthesized using an established precipitation system and characterized using transmission electron microscopy and 1.2% agarose gel electrophoresis. Biocompatibility and transfection of the nanoplexes in fibroblasts were evaluated using cytotoxicity assay and florescence microscopy, respectively. Polymerase chain reaction and enzyme-linked immunosorbent assay were performed to evaluate PDGF-B transfection after different time points of treatments, and the functionality of PDGF-B transfection was evaluated using the cell proliferation assay. Synthesized NCaPP nanoplexes incorporating the genes of GFP and PDGF-B were spherical in shape and measured about 30 to 50 nm in diameter. Gel electrophoresis confirmed DNA incorporation and stability within the nanoplexes, and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium reagent assay demonstrated their biocompatibility in fibroblasts. In vitro transfection studies revealed a higher and longer lasting transfection after NCaPP-PDGF-B treatment, which lasted up to 96 hours. Significantly enhanced fibroblast proliferation observed in NCaPP-PDGF-B-treated cells confirmed the functionality of these nanoplexes. NCaPP demonstrated higher levels of biocompatibility and efficiently transfected PDGF plasmids into fibroblasts under described in vitro conditions.

  20. Sugar in Infants, Children and Adolescents

    DEFF Research Database (Denmark)

    Mis, Nataša Fidler; Braegger, Christian; Bronsky, Jiri

    2017-01-01

    The consumption of sugars, particularly sugar-sweetened beverages (SSBs; beverages or drinks that contain added caloric sweeteners (i.e. sucrose, high-fructose corn syrup, fruit-juice concentrates), in European children and adolescents exceeds current recommendations. This is of concern because...... there is no nutritional requirement for free sugars, and infants have an innate preference for sweet taste, which may be modified and reinforced by pre- and postnatal exposures. Sugar containing beverages/free sugars increase the risk for overweight/obesity and dental caries, can result in poor nutrient supply...... and reduced dietary diversity and may be associated with increased risk of type 2 diabetes mellitus, cardiovascular risk, and other health effects. The term 'free sugars', includes all monosaccharides/disaccharides added to foods/beverages by the manufacturer/cook/consumer, plus sugars naturally present...

  1. Sugar in infants, children and adolescents

    DEFF Research Database (Denmark)

    Mis, Nataša Fidler; Braegger, Christian; Bronsky, Jiri

    2017-01-01

    The consumption of sugars, particularly sugar-sweetened beverages (SSBs; beverages or drinks that contain added caloric sweeteners (i.e. sucrose, high-fructose corn syrup, fruit-juice concentrates), in European children and adolescents exceeds current recommendations. This is of concern because...... there is no nutritional requirement for free sugars, and infants have an innate preference for sweet taste, which may be modified and reinforced by pre- and postnatal exposures. Sugar containing beverages/free sugars increase the risk for overweight/obesity and dental caries, can result in poor nutrient supply...... and reduced dietary diversity and may be associated with increased risk of type 2 diabetes mellitus, cardiovascular risk, and other health effects. The term 'free sugars', includes all monosaccharides/disaccharides added to foods/beverages by the manufacturer/cook/consumer, plus sugars naturally present...

  2. Acid-producing capacity from sugars and sugar alcohols among Lactobacillus isolates collected in connection with radiation therapy.

    Science.gov (United States)

    Almståhl, Annica; Rudbäck, Helena; Basic, Amina; Carlén, Anette; Alstad, Torgny

    2017-12-01

    To investigate the acid-producing capacity from sugars and sugar alcohols of oral Lactobacillus collected in connection with radiation therapy (RT) to the head and neck region. Lactobacillus were collected from the tongue, buccal mucosa and supragingival plaque in 24 patients before, during, and after RT. The acid-producing capacity of Lactobacillus isolates (n=211) was analyzed using a colorimetric fermentation test in microtiter plates. Solutions containing 2% sugars (sucrose, glucose, fructose, lactose) or sugar-alcohols (sorbitol and xylitol) were used. After 24h of incubation, bacterial acid-producing capacity was determined as strong (pH6). Data regarding intake frequency of sugar-rich products and products with sugar-alcohols was collected. The highest acid-producing capacity using the sugars was seen for isolates collected during RT. Sorbitol was fermented to a higher extent during and post RT, especially among isolates from plaque. Lactobacillus fermenting xylitol showed the highest acid-producing capacity during RT (psugar-rich products or sugar-alcohol containing products and Lactobacillus acid-producing capacity, were found. The results suggest that Lactobacillus isolates, collected from the tongue, buccal mucosa and supragingival plaque, have a higher acid-producing capacity using sugars and sugar-alcohols during RT than one year post RT. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Optimization of calcium phosphate fine ceramic powders preparation

    Science.gov (United States)

    Sezanova, K.; Tepavitcharova, S.; Rabadjieva, D.; Gergulova, R.; Ilieva, R.

    2013-12-01

    The effect of biomimetic synthesis method, reaction medium and further precursor treatments on the chemical and phase composition, crystal size and morphology of calcium phosphates was examined. Nanosized calcium phosphate precursors were biomimetically precipitated by the method of continuous precipitation in three types of reaction media at pH 8: (i) SBF as an inorganic electrolyte system; (ii) organic (glycerine) modified SBF (volume ratio of 1:1); (iii) polymer (10 g/l xanthan gum or 10 g/l guar gum) modified SBF (volume ratio of 1:1). After maturation (24 h) the samples were lyophilized, calcinated at 300°C for 3 hours, and washed with water, followed by new gelation, lyophilization and step-wise (200, 400, 600, 800, and 1000°C, each for 3 hours) sintering. The reaction medium influenced the chemical composition and particle size but not the morphology of the calcium phosphate powders. In all studied cases bi-phase calcium phosphate fine powders with well-shaped spherical grains, consisting of β-tricalcium phosphate (β-TCP) and hydroxyapatite (HA) with a Ca/P ratio of 1.3 - 1.6 were obtained. The SBF modifiers decreased the particle size of the product in the sequence guar gum ˜ xanthan gum < glycerin < SBF medium.

  4. Dynamic Changes in Yeast Phosphatase Families Allow for Specialization in Phosphate and Thiamine Starvation.

    Science.gov (United States)

    Nahas, John V; Iosue, Christine L; Shaik, Noor F; Selhorst, Kathleen; He, Bin Z; Wykoff, Dennis D

    2018-05-10

    Convergent evolution is often due to selective pressures generating a similar phenotype. We observe relatively recent duplications in a spectrum of Saccharomycetaceae yeast species resulting in multiple phosphatases that are regulated by different nutrient conditions - thiamine and phosphate starvation. This specialization is both transcriptional and at the level of phosphatase substrate specificity. In Candida glabrata , loss of the ancestral phosphatase family was compensated by the co-option of a different histidine phosphatase family with three paralogs. Using RNA-seq and functional assays, we identify one of these paralogs, CgPMU3 , as a thiamine phosphatase. We further determine that the 81% identical paralog CgPMU2 does not encode thiamine phosphatase activity; however, both are capable of cleaving the phosphatase substrate, 1-napthyl-phosphate. We functionally demonstrate that members of this family evolved novel enzymatic functions for phosphate and thiamine starvation, and are regulated transcriptionally by either nutrient condition, and observe similar trends in other yeast species. This independent, parallel evolution involving two different families of histidine phosphatases suggests that there were likely similar selective pressures on multiple yeast species to recycle thiamine and phosphate. In this work, we focused on duplication and specialization, but there is also repeated loss of phosphatases, indicating that the expansion and contraction of the phosphatase family is dynamic in many Ascomycetes. The dynamic evolution of the phosphatase gene families is perhaps just one example of how gene duplication, co-option, and transcriptional and functional specialization together allow species to adapt to their environment with existing genetic resources. Copyright © 2018, G3: Genes, Genomes, Genetics.

  5. Fluorescence Resonance Energy Transfer Assay for High-Throughput Screening of ADAMTS1 Inhibitors

    Directory of Open Access Journals (Sweden)

    Guanhua Du

    2011-12-01

    Full Text Available A disintegrin and metalloprotease with thrombospondin type I motifs-1 (ADAMTS1 plays a crucial role in inflammatory joint diseases and its inhibitors are potential candidates for anti-arthritis drugs. For the purposes of drug discovery, we reported the development and validation of fluorescence resonance energy transfer (FRET assay for high-throughput screening (HTS of the ADAMTS1 inhibitors. A FRET substrate was designed for a quantitative assay of ADAMTS1 activity and enzyme kinetics studies. The assay was developed into a 50-µL, 384-well assay format for high throughput screening of ADAMTS1 inhibitors with an overall Z’ factor of 0.89. ADAMTS1 inhibitors were screened against a diverse library of 40,960 total compounds with the established HTS system. Four structurally related hits, naturally occurring compounds, kuwanon P, kuwanon X, albafuran C and mulberrofuran J, extracted from the Chinese herb Morus alba L., were identified for further investigation. The results suggest that this FRET assay is an excellent tool, not only for measurement of ADAMTS1 activity but also for discovery of novel ADAMTS1 inhibitors with HTS.

  6. Injectable TEMPO-oxidized nanofibrillated cellulose/biphasic calcium phosphate hydrogel for bone regeneration.

    Science.gov (United States)

    Safwat, Engie; Hassan, Mohammad L; Saniour, Sayed; Zaki, Dalia Yehia; Eldeftar, Mervat; Saba, Dalia; Zazou, Mohamed

    2018-05-01

    Nanofibrillated cellulose, obtained from rice straw agricultural wastes was used as a substrate for the preparation of a new injectable and mineralized hydrogel for bone regeneration. Tetramethyl pyridine oxyl (TEMPO) oxidized nanofibrillated cellulose, was mineralized through the incorporation of a prepared and characterized biphasic calcium phosphate at a fixed ratio of 50 wt%. The TEMPO-oxidized rice straw nanofibrillated cellulose was characterized using transmission electron microscopy, Fourier transform infrared, and carboxylic content determination. The injectability and viscosity of the prepared hydrogel were evaluated using universal testing machine and rheometer testing, respectively. Cytotoxicity and alkaline phosphatase level tests on osteoblast like-cells for in vitro assessment of the biocompatibility were investigated. Results revealed that the isolated rice straw nanofibrillated cellulose is a nanocomposite of the cellulose nanofibers and silica nanoparticles. Rheological properties of the tested materials are suitable for use as injectable material and of nontoxic effect on osteoblast-like cells, as revealed by the positive alkaline phosphate assay. However, nanofibrillated cellulose/ biphasic calcium phosphate hydrogel showed higher cytotoxicity and lower bioactivity test results when compared to that of nanofibrillated cellulose.

  7. Bioactivity of calcium phosphate bioceramic coating fabricated by laser cladding

    Science.gov (United States)

    Zhu, Yizhi; Liu, Qibin; Xu, Peng; Li, Long; Jiang, Haibing; Bai, Yang

    2016-05-01

    There were always strong expectations for suitable biomaterials used for bone regeneration. In this study, to improve the biocompatiblity of titanium alloy, calcium phosphate bioceramic coating was obtained by laser cladding technology. The microstructure, phases, bioactivity, cell differentiation, morphology and resorption lacunae were investigated by optical microscope (OM), x-ray diffraction (XRD), methyl thiazolyl tetrazolium (MTT) assay, tartrate-resistant acid phosphatase (TRAP) staining and scanning electronic microscope (SEM), respectively. The results show that bioceramic coating consists of three layers, which are a substrate, an alloyed layer and a ceramic layer. Bioactive phases of β-tricalcium phosphate (β-TCP) and hydroxyapatite (HA) were found in ceramic coating. Osteoclast precursors have excellent proliferation on the bioceramic surface. The bioceramics coating could be digested by osteoclasts, which led to the resorption lacunae formed on its surface. It revealed that the gradient bioceramic coating has an excellent bioactivity.

  8. Bioactivity of calcium phosphate bioceramic coating fabricated by laser cladding

    International Nuclear Information System (INIS)

    Zhu, Yizhi; Liu, Qibin; Xu, Peng; Li, Long; Jiang, Haibing; Bai, Yang

    2016-01-01

    There were always strong expectations for suitable biomaterials used for bone regeneration. In this study, to improve the biocompatiblity of titanium alloy, calcium phosphate bioceramic coating was obtained by laser cladding technology. The microstructure, phases, bioactivity, cell differentiation, morphology and resorption lacunae were investigated by optical microscope (OM), x-ray diffraction (XRD), methyl thiazolyl tetrazolium (MTT) assay, tartrate-resistant acid phosphatase (TRAP) staining and scanning electronic microscope (SEM), respectively. The results show that bioceramic coating consists of three layers, which are a substrate, an alloyed layer and a ceramic layer. Bioactive phases of β-tricalcium phosphate (β-TCP) and hydroxyapatite (HA) were found in ceramic coating. Osteoclast precursors have excellent proliferation on the bioceramic surface. The bioceramics coating could be digested by osteoclasts, which led to the resorption lacunae formed on its surface. It revealed that the gradient bioceramic coating has an excellent bioactivity. (letter)

  9. Removal of phosphate from solution by adsorption and precipitation of calcium phosphate onto monohydrocalcite.

    Science.gov (United States)

    Yagi, Shintaro; Fukushi, Keisuke

    2012-10-15

    The sorption behavior and mechanism of phosphate on monohydrocalcite (CaCO(3)·H(2)O: MHC) were examined using batch sorption experiments as a function of phosphate concentrations, ionic strengths, temperatures, and reaction times. The mode of PO(4) sorption is divisible into three processes depending on the phosphate loading. At low phosphate concentrations, phosphate is removed by coprecipitation of phosphate during the transformation of MHC to calcite. The sorption mode at the low-to-moderate phosphate concentrations is most likely an adsorption process because the sorption isotherm at the conditions can be fitted reasonably with the Langmuir equation. The rapid sorption kinetics at the conditions is also consistent with the adsorption reaction. The adsorption of phosphate on MHC depends strongly on ionic strength, but slightly on temperature. The maximum adsorption capacities of MHC obtained from the regression of the experimental data to the Langmuir equation are higher than those reported for stable calcium carbonate (calcite or aragonite) in any conditions. At high phosphate concentrations, the amount of sorption deviates from the Langmuir isotherm, which can fit the low-to-moderate phosphate concentrations. Speciation-saturation analyses of the reacted solutions at the conditions indicated that the solution compositions which deviate from the Langmuir equation are supersaturated with respect to a certain calcium phosphate. The obtained calcium phosphate is most likely amorphous calcium phosphate (Ca(3)(PO(4))(2)·xH(2)O). The formation of the calcium phosphate depends strongly on ionic strength, temperature, and reaction times. The solubility of MHC is higher than calcite and aragonite because of its metastability. Therefore, the higher solubility of MHC facilitates the formation of the calcium phosphates more than with calcite and aragonite. Copyright © 2012 Elsevier Inc. All rights reserved.

  10. Sugar-free medicines are counterproductive.

    Science.gov (United States)

    Sundar, S

    2012-09-01

    Sugar in food and drinks is responsible for the poor dental health of many children and adults. On the other hand, there is no evidence that the small amount of sugar in medicines has been responsible for any dental problems. A recent British Heart Foundation survey found that nearly one in three UK children are eating sweets, chocolate and crisps three or more times a day. Hence it is futile administering sugar-free medicine to a child consuming lot of sweets. Moreover, sugar in medicines makes them palatable and bitter medicines inevitably affect compliance with the prescribed treatment. Poor compliance leads to inadequate treatment of illness and consequently increases the risk of complications from illness. Hence sugar-free medicines promoted as a public health policy could have actually caused more harm than any meaningful net benefit. There is an urgent need for a healthy debate and a fresh look at the policy of promoting sugar-free medicines.

  11. Solubility data and modeling for sugar alcohols in ionic liquids

    International Nuclear Information System (INIS)

    Okuniewski, Marcin; Ramjugernath, Deresh; Naidoo, Paramespri; Domańska, Urszula

    2014-01-01

    Highlights: • Solubility of D-sorbitol and xylitol in six ILs. • The (liquid + liquid) phase equilibrium of (SA + IL) with UCST. • Interesting properties of [BMIM][TDI] IL. • The correlation with NRTL model. - Abstract: Ionic liquids (ILs) are novel media characterized by strong interactions with different organic substances which leads to a wide spectrum of applications involving extraction. Ionic liquids have been used as a solvent for sugar alcohols, sugars and hydrates. This work demonstrates the experimental and theoretical study of (liquid + liquid) phase equilibria for two sugar alcohols, D-sorbitol and xylitol in a few ILs based on different cations and anions (namely, 1-ethyl-1-methylpiperidinium bis(trifluoromethylsulfonyl)imide [EMPIP][NTf 2 ], 1-hexyl-1-methylpiperidinium bis(trifluoromethylsulfonyl)imide [HMPIP][NTf 2 ], N-hexylquinolinium bis(trifluoromethylsulfonyl)imide [HQuin][NTf 2 ], N-hexylisoquinolinium bis(trifluoromethylsulfonyl)imide [HiQuin][NTf 2 ], 1-butyl-1-methylimidazolium 4,5-dicyano-2-(trifluoromethyl)-imidazolide [BMIM][TDI] and 1-(cyanomethyl)-3-methylimidazolium 4,5-dicyano-2-(trifluoromethyl)-imidazolide [CCNMIM][TDI]). This study was conducted to assess the applicability of the studied ILs for dissolution of these biomass-related materials. (Liquid + liquid) phase equilibrium diagrams (LLE) in binary systems (sugar alcohol + ionic liquid) were measured using the dynamic technique. The influence of the chemical structure of both the ionic liquids and sugar alcohols were established and is discussed

  12. Base-free Pd/TOMPP-Catalyzed Telomerization of 1,3-Butadiene with Carbohydrates and sugar alcohols

    NARCIS (Netherlands)

    Hausoul, P.J.C.; Bruijnincx, P.C.A.; Klein Gebbink, R.J.M.; Weckhuysen, B.M.

    2009-01-01

    Sugar and alcohol - a superior combo: The telomerization activity of the Pd/TOMPP catalyst is screened using thirteen different biomass-derived carbohydrates and sugar alcohols. High substrate conversions are achieved by using low Pd loading and without the use of an added base. In terms of

  13. Respiration-Dependent Utilization of Sugars in Yeasts: a Determinant Role for Sugar Transporters

    OpenAIRE

    Goffrini, Paola; Ferrero, Iliana; Donnini, Claudia

    2002-01-01

    In many yeast species, including Kluyveromyces lactis, growth on certain sugars (such as galactose, raffinose, and maltose) occurs only under respiratory conditions. If respiration is blocked by inhibitors, mutation, or anaerobiosis, growth does not take place. This apparent dependence on respiration for the utilization of certain sugars has often been suspected to be associated with the mechanism of the sugar uptake step. We hypothesized that in many yeast species, the permease activities fo...

  14. Pectinase activity determination: an early deceleration in the release of reducing sugars throws a spanner in the works!

    Directory of Open Access Journals (Sweden)

    Alessandra Biz

    Full Text Available Recently, it has been suggested that pectinases could be used to hydrolyze pectin in biorefineries based on pectin-rich agro-industrial wastes. However, for this to be viable, the cost of their production would need to be lowered significantly. In fact, over the last few decades, there have been many attempts to improve pectinase production by existing strains or to screen for new strains from environmental isolates. In these studies, it is necessary to measure pectinase activities. Many researchers use single-time-point assays that involve incubation of pectinolytic extracts with pectic substrates for a fixed time, followed by determination of the liberated reducing sugars. However, different researchers use quite different conditions for this assay. Furthermore, no attention has been given to the reaction profile during the assay. In the current work, we show, for the first time, that a significant deceleration of the rate of liberation of reducing sugars occurs over the first ten minutes of the reaction. As a consequence, the incubation time used in a single-time-point assay has a large effect on the value obtained for the activity. In fact, we demonstrate that, depending on the particular combination of incubation time, pectin concentration and reaction temperature, the same extract could be reported to have activities that differ by an order of magnitude. In addition, we show that the relative activities obtained with polygalacturonic acid do not correlate with those obtained with pectin. We conclude that it is currently impossible to make meaningful comparisons between pectinase activities reported in the literature by workers who have used different assay conditions. Therefore there is an urgent need for the development of a standardized assay for evaluating the saccharification potential of pectinase complexes.

  15. Sugar and Sugar Derivatives in Residues Produced from the UV Irradiation of Astrophysical Ice Analogs

    Science.gov (United States)

    Nuevo, M.; Sandford, S. A.; Cooper, G.

    2016-01-01

    A large variety and number of organic compounds of prebiotic interest are known to be present in carbonaceous chondrites. Among them, one sugar (dihydroxyacetone) as well as several sugar acids, sugar alcohols, and other sugar derivatives have been reported in the Murchison and Murray meteorites. Their presence, along with amino acids, amphiphiles, and nucleobases strongly suggests that molecules essential to life can form abiotically under astrophysical conditions. This hypothesis is supported by laboratory studies on the formation of complex organic molecules from the ultraviolet (UV) irradiation of simulated astrophysical ice mixtures consisting of H2O, CO, CO2, CH3OH, CH4, NH3, etc., at low temperature. In the past 15 years, these studies have shown that the organic residues recovered at room temperature contain amino acids, amphiphiles, nucleobases, as well as other complex organics. However, no systematic search for the presence of sugars and sugar derivatives in laboratory residues have been reported to date, despite the fact that those compounds are of primary prebiotic significance. Indeed, only small (up to 3 carbon atoms) sugar derivatives including glycerol and glyceric acid have been detected in residues so far.

  16. Sphingosine-1-phosphate (S1P) enhances glomerular endothelial cells activation mediated by anti-myeloperoxidase antibody-positive IgG.

    Science.gov (United States)

    Sun, Xiao-Jing; Chen, Min; Zhao, Ming-Hui

    2018-03-01

    Cumulating evidences suggested an important role of sphingosine-1-phosphate (S1P) and its receptors in regulating endothelial barrier integrity. Our previous study revealed that the circulating S1P levels and renal expression of S1PRs correlated with disease activity and renal damage in patients with antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV). This study investigated the role of S1P and its receptors in myeloperoxidase (MPO)-ANCA-positive IgG-mediated glomerular endothelial cell (GEnC) activation. The effect of S1P on morphological alteration of GEnCs in the presence of MPO-ANCA-positive IgG was observed. Permeability assay was performed to determine endothelial monolayer activation in quantity. Both membrane-bound and soluble ICAM-1 and VCAM-1 levels were measured. Furthermore, antagonists and/or agonists of various S1PRs were employed to determine the role of different S1PRs. S1P enhanced MPO-ANCA-positive IgG-induced disruption of tight junction and disorganization of cytoskeleton in GEnCs. S1P induced further increase in monolayer permeability of GEnC monolayers in the presence of MPO-ANCA-positive IgG. S1P enhanced MPO-ANCA-positive IgG-induced membrane-bound and soluble ICAM-1/VCAM-1 up-regulation of GEnCs. Soluble ICAM-1 levels in the supernatants of GEnCs stimulated by S1P and MPO-ANCA-positive IgG increased upon pre-incubation of S1PR1 antagonist, while pre-incubation of GEnCs with the S1PR1 agonist down-regulated sICAM-1 level. Blocking S1PR2-4 reduced sICAM-1 levels in the supernatants of GEnCs stimulated by S1P and MPO-ANCA-positive IgG. Pre-incubation with S1PR5 agonist could increase sICAM-1 level in the supernatants of GEnC stimulated by S1P and MPO-ANCA-positive IgG. S1P can enhance MPO-ANCA-positive IgG-mediated GEnC activation through S1PR2-5. © 2017 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

  17. Lipopolysaccharides of the cyanobacterium Microcystis aeruginosa

    Energy Technology Data Exchange (ETDEWEB)

    Raziuddin, S.; Siegelman, H.W.; Tornabene, T.G.

    1983-01-01

    Lipopolysaccharides (LPS) of two isolates of Microcystis aeruginosa were extracted with phenol/water and purified. Cesium chloride gradient ultracentrifugation of these preparations yielded only one fraction. The LPS contained significant amounts of 3-deoxy-D-manno-octulosonic acid, glucose, 3-deoxy sugars, glucosamine, fatty acids, fatty acid esters, hexoses, and phosphate. Heptose, a characteristic sugar component of the polysaccharide moiety of LPS of most gram-negative bacteria was absent. Lipopolysaccharides and lipid A hydrolysate of LPS preparations were active in mouse lethality and Limulus lysate gelation. The lipid A moiety was slightly less active in toxicity and Limulus lysate gelation assay than the intact LPS. The LPS and lipid A moiety of the two isolates of M. aeruginosa were less active in toxicity in mice and Limulus test than LPS of Salmonella abortus equi. 37 references, 1 figure, 3 tables.

  18. Technical Report for DE-FG02-03ER46029 Sugar-Coated PPEs, Novel Nanomaterials and Sensing Modules for Disease and Bioterrorism Related Threats

    Energy Technology Data Exchange (ETDEWEB)

    Uwe Bunz

    2003-08-27

    The detection and sensing of biological warfare agents (Ricin, Anthrax toxin), of disease agents (cholera, botulinum and tetanus toxins, influenza virus etc) and of biologically active species is important for national security and disease control. A premiere goal would be the simple colorimetric or fluorimetric detection of such toxins by a dipstick test. It would be desirable to sense 5,000-10,000 toxin molecules, i.e. 10-100 fg of a toxin contained 1-5 mL of sample. Fluorescent conjugated polymers should be particularly interesting in this regard, because they can carry multiple identical and/or different recognition units. Such an approach is particularly valuable for the detection of lectin toxins, because these bind to oligomeric carbohydrate displays. Lectins bind multivalently to sugars, i.e. several covalently connected sugar moieties have to be exposed to the lectin at the same time to obtain binding. The requirement of multivalency of the lectin-sugar interactions should allow a very sensitive detection of lectins with sugar coated conjugated polymers in an agglutination type assay, where the fluorescence of the PPEs disappears upon binding to the lectins. High molecular weights of the used PPEs would mean high sensitivity. Herein we present our progress towards that goal up to date.

  19. A free sugars daily value (DV) identifies more "less healthy" prepackaged foods and beverages than a total sugars DV.

    Science.gov (United States)

    Bernstein, Jodi T; Labonté, Marie-Ève; Franco-Arellano, Beatriz; Schermel, Alyssa; L'Abbé, Mary R

    2018-04-01

    Regulatory changes in Canada will require food labels to have a benchmark [% Daily Value, %DV] for total sugars, based on 100 g/day, while US labels will require a %DV for added sugars, based on 50 g/day. The objective of this study was to compare two labelling policies, a total sugars DV (100 g/day) and a free sugars DV (50 g/day) on food labels. This cross-sectional analysis of the Food Label Information Program database focussed on top sources of total sugars intake in Canada (n = 6924 foods). Products were categorized as "less healthy" using two sets of criteria: a) free sugars levels exceeding the WHO guidelines (≥10% energy from free sugars); and b) exceeding healthfulness cut-offs of the Food Standards Australia New Zealand Nutrient Profiling Scoring Criterion (FSANZ-NPSC). The proportion of "less healthy" products with ≥15%DV (defined as "a lot" of sugars i.e. high in sugars, based on Health Canada's %DV labelling footnote and educational message for dietary guidance) were compared for each sugar labelling scenario. The free sugars DV showed better alignment with both methods for assessing "healthfulness" than the total sugars DV. The free sugars DV identified a greater proportion of "less healthy" foods with ≥15%DV, based on both the FSANZ-NPSC (70% vs. 45%, p chocolate bars, confectionery, and frozen desserts categories. Compared to total sugars DV labelling, using a free sugars DV identified more "less healthy" foods. Findings support the adoption of free sugars labelling. Copyright © 2018 Elsevier Inc. All rights reserved.

  20. Aromatics extraction from pyrolytic sugars using ionic liquid to enhance sugar fermentability

    NARCIS (Netherlands)

    Li, X.; Luque-Moreno, L.C.; Oudenhoven, Stijn; Rehmann, L.; Kersten, Sascha R.A.; Schuur, Boelo

    2016-01-01

    Fermentative bioethanol production from pyrolytic sugars was improved via aromatics removal by liquid–liquid extraction. As solvents, the ionic liquid (IL) trihexyltetradecylphosphonium dicyanamide (P666,14[N(CN)2]) and ethyl acetate (EA) were compared. Two pyrolytic sugar solutions were created