Characterization and modification of enzymes in the 2-ketoisovalerate biosynthesis pathway of Ralstonia eutropha H16

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lu, JN; Brigham, CJ; Plassmeier, JK

2014-08-01

2-Ketoisovalerate is an important cellular intermediate for the synthesis of branched-chain amino acids as well as other important molecules, such as pantothenate, coenzyme A, and glucosinolate. This ketoacid can also serve as a precursor molecule for the production of biofuels, pharmaceutical agents, and flavor agents in engineered organisms, such as the betaproteobacterium Ralstonia eutropha. The biosynthesis of 2-ketoisovalerate from pyruvate is carried out by three enzymes: acetohydroxyacid synthase (AHAS, encoded by ilvBH), acetohydroxyacid isomeroreductase (AHAIR, encoded by ilvC), and dihydroxyacid dehydratase (DHAD, encoded by ilvD). In this study, enzymatic activities and kinetic parameters were determined for each of the threemore » R. eutropha enzymes as heterologously purified proteins. AHAS, which serves as a gatekeeper for the biosynthesis of all three branched-chain amino acids, demonstrated the tightest regulation through feedback inhibition by l-valine (IC50 = 1.2 mM), l-isoleucine (IC50 = 2.3 mM), and l-leucine (IC50 = 5.4 mM). Intermediates in the valine biosynthesis pathway also exhibit feedback inhibitory control of the AHAS enzyme. In addition, AHAS has a very weak affinity for pyruvate (K-M = 10.5 mu M) and is highly selective towards 2-ketobutyrate (R = 140) as a second substrate. AHAIR and DHAD are also inhibited by the branched-chain amino acids, although to a lesser extent when compared to AHAS. Experimental evolution and rational site-directed mutagenesis revealed mutants of the regulatory subunit of AHAS (IlvH) (N11S, T34I, A36V, T104S, N11F, G14E, and N29H), which, when reconstituted with wild-type IlvB, lead to AHAS having reduced valine, leucine, and isoleucine sensitivity. The study of the kinetics and inhibition mechanisms of R. eutropha AHAS, AHAIR, and DHAD has shed light on interactions between these enzymes and the products they produce; it, therefore, can be used to engineer R. eutropha strains with optimal production of 2-ketoisovalerate for value-added materials.« less

Engineering of Ralstonia eutropha for production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) from fructose and solid-state properties of the copolymer.

PubMed

Fukui, Toshiaki; Abe, Hideki; Doi, Yoshiharu

2002-01-01

Recombinant Ralstonia eutropha capable of producing poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) copolymer [P(3HB-co-3HHx)] from fructose was engineered by introduction of genes for crotonyl-CoA reductase (CCR) from Streptomyces cinnamonensis (ccrSc) and for PHA synthase and (R)-specific enoyl-CoA hydratase from Aeromonas caviae (phaC-JAc). In this recombinant strain, C6-acyl-CoA intermediates were provided via beta-ketothiolase-mediated elongation of butyryl-CoA, which was generated from crotonyl-CoA by the function of CCR. The recombinant strain could accumulate the copolyester up to 48 wt % of dry cell weight with 1.5 mol % of 3HHx fraction from fructose, when the expression of ccrSc under the control of the PBAD promoter was induced with 0.01% L-arabinose. The absence of L-arabinose or the deletion of ccrSc from the plasmid resulted in accumulation of poly(3-hydroxybutyrate) homopolymer, indicating the critical role of CCR in the formation of the 3-hydroxyhexanoate unit. Higher CCR activity obtained by the addition of a larger amount of L-arabinose did not affect the composition but reduced the intracellular content of the copolyester. The P(3HB-co-1.5 mol % 3HHx) copolyester produced from fructose by the recombinant R. eutropha showed relatively lower melting temperatures (150 degrees C and 161 degrees C) and lower crystallinity (48 +/- 5%) compared to those (175 degrees C and 60 +/- 5%) of P(3HB) homopolymer. It has been found that the incorporation of a small amount (1.5 mol %) of 3HHx units into P(3HB) sequences leads to a remarkable change in the solid-state properties of P(3HB) crystals. The present study demonstrates the potential of the engineered pathway for the production of copolyesters having favorable characteristics from inexpensive carbon resources.

A closer look on the polyhydroxybutyrate- (PHB-) negative phenotype of Ralstonia eutropha PHB-4.

PubMed

Raberg, Matthias; Voigt, Birgit; Hecker, Michael; Steinbüchel, Alexander

2014-01-01

The undefined poly(3-hydroxybutyrate)- (PHB-) negative mutant R. eutropha PHB-4 was generated in 1970 by 1-nitroso-3-nitro-1-methylguanidine (NMG) treatment. Although being scientific relevant, its genotype remained unknown since its isolation except a recent first investigation. In this study, the mutation causing the PHA-negative phenotype of R. eutropha PHB-4 was confirmed independently: sequence analysis of the phaCAB operon identified a G320A mutation in phaC yielding a stop codon, leading to a massively truncated PhaC protein of 106 amino acids (AS) in R. eutropha PHB-4 instead of 589 AS in the wild type. No other mutations were observed within the phaCAB operon. As further mutations probably occurred in the genome of mutant PHB-4 potentially causing secondary effects on the cells' metabolism, the main focus of the study was to perform a 2D PAGE-based proteome analysis in order to identify differences in the proteomes of the wild type and mutant PHB-4. A total of 20 differentially expressed proteins were identified which provide valuable insights in the metabolomic changes of mutant PHB-4. Besides excretion of pyruvate, mutant PHB-4 encounters the accumulation of intermediates such as pyruvate and acetyl-CoA by enhanced expression of the observed protein species: (i) ThiJ supports biosynthesis of cofactor TPP and thereby reinforces the 2-oxoacid dehydrogenase complexes as PDHC, ADHC and OGDHC in order to convert pyruvate at a higher rate and the (ii) 3-isopropylmalate dehydrogenase LeuB3 apparently directs pyruvate to synthesis of several amino acids. Different (iii) acylCoA-transferases enable transfer reactions between organic acid intermediates, and (iv) citrate lyase CitE4 regenerates oxaloacetate from citrate for conversion with acetyl-CoA in the TCC in an anaplerotic reaction. Substantial amounts of reduction equivalents generated in the TCC are countered by (v) synthesis of more ubiquinones due to enhanced synthesis of MenG2 and MenG3, thereby improving the respiratory chain which accepts electrons from NADH and succinate.

A Closer Look on the Polyhydroxybutyrate- (PHB-) Negative Phenotype of Ralstonia eutropha PHB-4

PubMed Central

Raberg, Matthias; Voigt, Birgit; Hecker, Michael; Steinbüchel, Alexander

2014-01-01

The undefined poly(3-hydroxybutyrate)- (PHB-) negative mutant R. eutropha PHB-4 was generated in 1970 by 1-nitroso-3-nitro-1-methylguanidine (NMG) treatment. Although being scientific relevant, its genotype remained unknown since its isolation except a recent first investigation. In this study, the mutation causing the PHA-negative phenotype of R. eutropha PHB-4 was confirmed independently: sequence analysis of the phaCAB operon identified a G320A mutation in phaC yielding a stop codon, leading to a massively truncated PhaC protein of 106 amino acids (AS) in R. eutropha PHB-4 instead of 589 AS in the wild type. No other mutations were observed within the phaCAB operon. As further mutations probably occurred in the genome of mutant PHB-4 potentially causing secondary effects on the cells' metabolism, the main focus of the study was to perform a 2D PAGE-based proteome analysis in order to identify differences in the proteomes of the wild type and mutant PHB-4. A total of 20 differentially expressed proteins were identified which provide valuable insights in the metabolomic changes of mutant PHB-4. Besides excretion of pyruvate, mutant PHB-4 encounters the accumulation of intermediates such as pyruvate and acetyl-CoA by enhanced expression of the observed protein species: (i) ThiJ supports biosynthesis of cofactor TPP and thereby reinforces the 2-oxoacid dehydrogenase complexes as PDHC, ADHC and OGDHC in order to convert pyruvate at a higher rate and the (ii) 3-isopropylmalate dehydrogenase LeuB3 apparently directs pyruvate to synthesis of several amino acids. Different (iii) acylCoA-transferases enable transfer reactions between organic acid intermediates, and (iv) citrate lyase CitE4 regenerates oxaloacetate from citrate for conversion with acetyl-CoA in the TCC in an anaplerotic reaction. Substantial amounts of reduction equivalents generated in the TCC are countered by (v) synthesis of more ubiquinones due to enhanced synthesis of MenG2 and MenG3, thereby improving the respiratory chain which accepts electrons from NADH and succinate. PMID:24787649

Genome-Based Analysis and Gene Dosage Studies Provide New Insight into 3-Hydroxy-4-Methylvalerate Biosynthesis in Ralstonia eutropha

PubMed Central

Ushimaru, Kazunori; Mizuno, Shoji

2015-01-01

Recombinant Ralstonia eutropha strain PHB−4 expressing the broad-substrate-specificity polyhydroxyalkanoate (PHA) synthase 1 from Pseudomonas sp. strain 61-3 (PhaC1Ps) synthesizes a PHA copolymer containing the branched side-chain unit 3-hydroxy-4-methylvalerate (3H4MV), which has a carbon backbone identical to that of leucine. Mutant strain 1F2 was derived from R. eutropha strain PHB−4 by chemical mutagenesis and shows higher levels of 3H4MV production than does the parent strain. In this study, to understand the mechanisms underlying the enhanced production of 3H4MV, whole-genome sequencing of strain 1F2 was performed, and the draft genome sequence was compared to that of parent strain PHB−4. This analysis uncovered four point mutations in the 1F2 genome. One point mutation was found in the ilvH gene at amino acid position 36 (A36T) of IlvH. ilvH encodes a subunit protein that regulates acetohydroxy acid synthase III (AHAS III). AHAS catalyzes the conversion of pyruvate to 2-acetolactate, which is the first reaction in the biosynthesis of branched amino acids such as leucine and valine. Thus, the A36T IlvH mutation may show AHAS tolerance to feedback inhibition by branched amino acids, thereby increasing carbon flux toward branched amino acid and 3H4MV biosynthesis. Furthermore, a gene dosage study and an isotope tracer study were conducted to investigate the 3H4MV biosynthesis pathway. Based on the observations in these studies, we propose a 3H4MV biosynthesis pathway in R. eutropha that involves a condensation reaction between isobutyryl coenzyme A (isobutyryl-CoA) and acetyl-CoA to form the 3H4MV carbon backbone. PMID:25645560

A process economic assessment of hydrocarbon biofuels production using chemoautotrophic organisms

DOE Office of Scientific and Technical Information (OSTI.GOV)

Khan, NE; Myers, JA; Tuerk, AL

Economic analysis of an ARPA-e Electrofuels (http://arpa-e.energy.gov/?q=arpa-e-programs/electrofuels) process is presented, utilizing metabolically engineered Rhodobacter capsulatus or Ralstonia eutropha to produce the C30+ hydrocarbon fuel, botryococcene, from hydrogen, carbon dioxide, and oxygen. The analysis is based on an Aspen plus (R) bioreactor model taking into account experimentally determined Rba. capsulatus and Rls. eutropha growth and maintenance requirements, reactor residence time, correlations for gas-liquid mass-transfer coefficient, gas composition, and specific cellular fuel productivity. Based on reactor simulation results encompassing technically relevant parameter ranges, the capital and operating costs of the process were estimated for 5000 bbl-fuel/day plant and used to predict fuelmore » cost. Under the assumptions used in this analysis and crude oil prices, the Levelized Cost of Electricity (LCOE) required for economic feasibility must be less than 2(sic)/kWh. While not feasible under current market prices and costs, this work identifies key variables impacting process cost and discusses potential alternative paths toward economic feasibility. (C) 2014 Elsevier Ltd. All rights reserved.« less

New Insights into PhaM-PhaC-Mediated Localization of Polyhydroxybutyrate Granules in Ralstonia eutropha H16

PubMed Central

Bresan, Stephanie

2017-01-01

ABSTRACT The formation and localization of polyhydroxybutyrate (PHB) granules in Ralstonia eutropha are controlled by PhaM, which interacts both with the PHB synthase (PhaC) and with the bacterial nucleoid. Here, we studied the importance of proline and lysine residues of two C-terminal PAKKA motifs in PhaM for their importance in attaching PHB granules to DNA by in vitro and in vivo methods. Substitution of the lysine residues but not of the proline residues resulted in detachment of formed PHB granules from the nucleoid. Instead, formation of PHB granule clusters at polar regions of the rod-shaped cells and an unequal distribution of PHB granules to daughter cells were observed. The formation of PHB granules was studied by the expression of chromosomally anchored gene fusions of fluorescent proteins with PhaM and PhaC in different backgrounds. PhaM and PhaC fusions showed a distinct colocalization at formed PHB granules in the nucleoid region of the wild type. In a ΔphaC background, PhaM and the catalytically inactive PhaCC319A protein were not able to form fluorescent foci, indicating that correct positioning requires the formation of PHB. Furthermore, time-lapse experiments revealed that PhaC and PhaM proteins detach from formed PHB granules at later stages, resulting in a nonhomogeneous population of PHB granules. This could explain why growth of individual PHB granules stops under PHB-permissive conditions at a certain size. IMPORTANCE PHB granules are storage compounds for carbon and energy in many prokaryotes. Equal distribution of accumulated PHB granules during cell division is therefore important for optimal fitness of the daughter cells. In R. eutropha, PhaM is responsible for maximal activity of PHB synthase, for initiation of PHB granule formation at discrete regions in the cells, and for association of formed PHB granules with the nucleoid. Here we found that four lysine residues of C-terminal PhaM sequence motifs are essential for association of PHB granules with the nucleoid. Furthermore, we followed PHB granule formation by time-lapse microscopy and provide evidence for aging of PHB granules that is manifested by detachment of previously PHB granule-associated PhaM and PHB synthase. PMID:28389545

Splice form variant and amino acid changes in MDR49 confers DDT resistance in transgenic Drosophila

PubMed Central

Seong, Keon Mook; Sun, Weilin; Clark, John M.; Pittendrigh, Barry R.

2016-01-01

The ATP-binding cassette (ABC) transporters represent a superfamily of proteins that have important physiological roles in both prokaryotes and eukaryotes. In insects, ABC transporters have previously been implicated in insecticide resistance. The 91-R strain of Drosophila melanogaster has been intensely selected with DDT over six decades. A recent selective sweeps analysis of 91-R implicated the potential role of MDR49, an ABC transporter, in DDT resistance, however, to date the details of how MDR49 may play a role in resistance have not been elucidated. In this study, we investigated the impact of structural changes and an alternative splicing event in MDR49 on DDT-resistance in 91-R, as compared to the DDT susceptible strain 91-C. We observed three amino acid differences in MDR49 when 91-R was compared with 91-C, and only one isoform (MDR49B) was implicated in DDT resistance. A transgenic Drosophila strain containing the 91-R-MDR49B isoform had a significantly higher LD50 value as compared to the 91-C-MDR49B isoform at the early time points (6 h to 12 h) during DDT exposure. Our data support the hypothesis that the MDR49B isoform, with three amino acid mutations, plays a role in the early aspects of DDT resistance in 91-R. PMID:27003579

Splice form variant and amino acid changes in MDR49 confers DDT resistance in transgenic Drosophila.

PubMed

Seong, Keon Mook; Sun, Weilin; Clark, John M; Pittendrigh, Barry R

2016-03-22

The ATP-binding cassette (ABC) transporters represent a superfamily of proteins that have important physiological roles in both prokaryotes and eukaryotes. In insects, ABC transporters have previously been implicated in insecticide resistance. The 91-R strain of Drosophila melanogaster has been intensely selected with DDT over six decades. A recent selective sweeps analysis of 91-R implicated the potential role of MDR49, an ABC transporter, in DDT resistance, however, to date the details of how MDR49 may play a role in resistance have not been elucidated. In this study, we investigated the impact of structural changes and an alternative splicing event in MDR49 on DDT-resistance in 91-R, as compared to the DDT susceptible strain 91-C. We observed three amino acid differences in MDR49 when 91-R was compared with 91-C, and only one isoform (MDR49B) was implicated in DDT resistance. A transgenic Drosophila strain containing the 91-R-MDR49B isoform had a significantly higher LD50 value as compared to the 91-C-MDR49B isoform at the early time points (6 h to 12 h) during DDT exposure. Our data support the hypothesis that the MDR49B isoform, with three amino acid mutations, plays a role in the early aspects of DDT resistance in 91-R.

Hydrogen-induced structural changes at the nickel site of the regulatory [NiFe] hydrogenase from Ralstonia eutropha detected by X-ray absorption spectroscopy.

PubMed

Haumann, Michael; Porthun, Antje; Buhrke, Thorsten; Liebisch, Peter; Meyer-Klaucke, Wolfram; Friedrich, Bärbel; Dau, Holger

2003-09-23

For the first time, the nickel site of the hydrogen sensor of Ralstonia eutropha, the regulatory [NiFe] hydrogenase (RH), was investigated by X-ray absorption spectroscopy (XAS) at the nickel K-edge. The oxidation state and the atomic structure of the Ni site were investigated in the RH in the absence (air-oxidized, RH(ox)) and presence of hydrogen (RH(+H2)). Incubation with hydrogen is found to cause remarkable changes in the spectroscopic properties. The Ni-C EPR signal, indicative of Ni(III), is detectable only in the RH(+H2) state. XANES and EXAFS spectra indicate a coordination of the Ni in the RH(ox) and RH(+H2) that pronouncedly differs from the one in standard [NiFe] hydrogenases. Also, the changes induced by exposure to H(2) are unique. A drastic modification in the XANES spectra and an upshift of the K-edge energy from 8339.8 (RH(ox)) to 8341.1 eV (RH(+H2)) is observed. The EXAFS spectra indicate a change in the Ni coordination in the RH upon exposure to H(2). One likely interpretation of the data is the detachment of one sulfur ligand in RH(+H2) and the binding of additional (O,N) or H ligands. The following Ni oxidation states and coordinations are proposed: five-coordinated Ni(II)(O,N)(2)S(3) for RH(ox) and six-coordinated Ni((III))(O,N)(3)X(1)S(2) [X being either an (O,N) or H ligand] for RH(+H2). Implications of the structural features of the Ni site of the RH in relation to its function, hydrogen sensing, are discussed.

Ralstonia eutropha H16 in progress: Applications beside PHAs and establishment as production platform by advanced genetic tools.

PubMed

Raberg, Matthias; Volodina, Elena; Lin, Kaichien; Steinbüchel, Alexander

2018-06-01

Ralstonia eutropha strain H16 is a Gram-negative non-pathogenic betaproteobacterium ubiquitously found in soils and has been the subject of intensive research for more than 50 years. Due to its remarkable metabolically versatility, it utilizes a broad range of renewable heterotrophic resources. The substrate utilization range can be further extended by metabolic engineering as genetic tools are available. It has become the best studied "Knallgas" bacterium capable of chemolithoautotrophic growth with hydrogen as the electron donor and carbon dioxide as the carbon source. It also serves as a model organism to study the metabolism of poly(β-hydroxybutyrate), a polyester which is accumulated within the cells for storage of both carbon and energy. Thermoplastic and biodegradable properties of this polyhydroxyalkanoate (PHA) have attracted much biotechnical interest as a replacement for fossil resource-based plastics. The first applications of R. eutropha aimed at chemolithoautotrophic production of single cell protein (SCP) for food and feed and the synthesis of various PHAs. The complete annotated genome is available allowing systematic biology approaches together with data provided by available omics studies. Besides PHAs, novel biopolymers of 2-hydroxyalkanoates and polythioesters or cyanophycin as well as chemicals such as alcohols, alkanes, alkenes, and further interesting value added chemicals significantly recently extended the range of products synthesized by R. eutropha. High cell density cultivations can be performed without too much effort and the available repertoire of genetic tools is rapidly growing. Altogether, this qualifies R. eutropha strain H16 to become a production platform strain for a large spectrum of products.

RubisCO selection using the vigorously aerobic and metabolically versatile bacterium Ralstonia eutropha.

PubMed

Satagopan, Sriram; Tabita, F Robert

2016-08-01

Recapturing atmospheric CO2 is key to reducing global warming and increasing biological carbon availability. Ralstonia eutropha is a biotechnologically useful aerobic bacterium that uses the Calvin-Benson-Bassham (CBB) cycle and the enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) for CO2 utilization, suggesting that it may be a useful host to bioselect RubisCO molecules with improved CO2 -capture capabilities. A host strain of R. eutropha was constructed for this purpose after deleting endogenous genes encoding two related RubisCOs. This strain could be complemented for CO2 -dependent growth by introducing native or heterologous RubisCO genes. Mutagenesis and suppressor selection identified amino acid substitutions in a hydrophobic region that specifically influences RubisCO's interaction with its substrates, particularly O2 , which competes with CO2 at the active site. Unlike most RubisCOs, the R. eutropha enzyme has evolved to retain optimal CO2 -fixation rates in a fast-growing host, despite the presence of high levels of competing O2 . Yet its structure-function properties resemble those of several commonly found RubisCOs, including the higher plant enzymes, allowing strategies to engineer analogous enzymes. Because R. eutropha can be cultured rapidly under harsh environmental conditions (e.g., with toxic industrial flue gas), in the presence of near saturation levels of oxygen, artificial selection and directed evolution studies in this organism could potentially impact efforts toward improving RubisCO-dependent biological CO2 utilization in aerobic environments. d-ribulose 1,5-bisphosphate carboxylase/oxygenase, EC 4.1.1.39; phosphoribulokinase, EC 2.7.1.19. © 2016 Federation of European Biochemical Societies.

Ralstonia eutropha H16 Flagellation Changes According to Nutrient Supply and State of Poly(3-Hydroxybutyrate) Accumulation▿

PubMed Central

Raberg, Matthias; Reinecke, Frank; Reichelt, Rudolf; Malkus, Ursula; König, Simone; Pötter, Markus; Fricke, Wolfgang Florian; Pohlmann, Anne; Voigt, Birgit; Hecker, Michael; Friedrich, Bärbel; Bowien, Botho; Steinbüchel, Alexander

2008-01-01

Two-dimensional polyacrylamide gel electrophoresis (2D PAGE), in combination with matrix-assisted laser desorption ionization-time of flight analysis, and the recently revealed genome sequence of Ralstonia eutropha H16 were employed to detect and identify proteins that are differentially expressed during different phases of poly(3-hydroxybutyric acid) (PHB) metabolism. For this, a modified protein extraction protocol applicable to PHB-harboring cells was developed to enable 2D PAGE-based proteome analysis of such cells. Subsequently, samples from (i) the exponential growth phase, (ii) the stationary growth phase permissive for PHB biosynthesis, and (iii) a phase permissive for PHB mobilization were analyzed. Among several proteins exhibiting quantitative changes during the time course of a cultivation experiment, flagellin, which is the main protein of bacterial flagella, was identified. Initial investigations that report on changes of flagellation for R. eutropha were done, but 2D PAGE and electron microscopic examinations of cells revealed clear evidence that R. eutropha exhibited further significant changes in flagellation depending on the life cycle, nutritional supply, and, in particular, PHB metabolism. The results of our study suggest that R. eutropha is strongly flagellated in the exponential growth phase and loses a certain number of flagella in transition to the stationary phase. In the stationary phase under conditions permissive for PHB biosynthesis, flagellation of cells admittedly stagnated. However, under conditions permissive for intracellular PHB mobilization after a nitrogen source was added to cells that are carbon deprived but with full PHB accumulation, flagella are lost. This might be due to a degradation of flagella; at least, the cells stopped flagellin synthesis while normal degradation continued. In contrast, under nutrient limitation or the loss of phasins, cells retained their flagella. PMID:18502919

Production and purification of a soluble hydrogenase from Ralstonia eutropha H16 for potential hydrogen fuel cell applications.

PubMed

Jugder, Bat-Erdene; Lebhar, Helene; Aguey-Zinsou, Kondo-Francois; Marquis, Christopher P

2016-01-01

The soluble hydrogenase (SH) from Ralstonia eutropha H16 is a promising candidate enzyme for H2-based biofuel application as it favours H2 oxidation and is relatively oxygen-tolerant. In this report, bioprocess development studies undertaken to produce and purify an active SH are described, based on the methods previously reported [1], [2], [3], [4]. Our modifications are: •Upstream method optimizations were undertaken on heterotrophic growth media and cell lysis involving ultrasonication.•Two anion exchangers (Q Sepharose and RESOURCE Q) and size exclusion chromatographic (Superdex 200) matrices were successfully employed for purification of a hexameric SH from R. eutropha.•The H2 oxidizing activity of the SH was demonstrated spectrophotometrically in solution and also immobilized on an EPG electrode using cyclic voltammetry.

Physiological-biochemical properties and the ability to synthesize polyhydroxyalkanoates of the glucose-utilizing strain of the hydrogen bacterium Ralstonia eutropha B8562.

PubMed

Volova, T G; Trusova, M Y; Kalacheva, G S; Kozhevnicov, I V

2006-11-01

Physiological-biochemical, genetic, and cultural properties of the glucose-utilizing mutant strain Ralstonia eutropha B8562 have been compared with those of its parent strain R. eutropha B5786. It has been shown that growth characteristics of the strain cultured on glucose as the sole carbon and energy source are comparable with those of the parent strain. Strain B8562 is characterized by high polyhydroxyalkanoate (PHA) yields on different carbon sources (CO(2), fructose, and glucose). PHA accumulation in the strain batch cultured on glucose under nitrogen deficiency reaches 90 %. The major monomer in the PHA is beta-hydroxybutyric acid (more than 99 mol %); the identified minor components are beta-hydroxyvaleric acid (0.25-0.72 mol %) and beta-hydroxyhexanoic acid (0.08-1.5 mol %). The strain is a promising PHA producer on available sugar-containing media with glucose.

PHA Productivity and Yield of Ralstonia eutropha When Intermittently or Continuously Fed a Mixture of Short Chain Fatty Acids

PubMed Central

Chakraborty, Panchali; Muthukumarappan, Kasiviswanathan; Gibbons, William R.

2012-01-01

The research described in this present study was part of a larger effort focused on developing a dual substrate, dual fermentation process to produce Polyhydroxyalkanoate (PHA). The focus of this study was developing and optimizing a strategy for feeding a mixture of SCFAs (simulated ARF) and maximizing PHA production in a cost-effective way. Three different feeding strategies were examined in this study. The substrate evaluated in this study for the growth phase of R. eutropha was condensed corn solubles, a low-value byproduct of the dry-mill, corn ethanol industry. The culture was grown to high cell densities in nitrogen-supplemented condensed corn solubles media in 5 L bioreactors. The overall growth rate of R. eutropha was 0.2 h−1. The 20 mL ARF feeding every 3 h from 48 to 109 h strategy gave the best results in terms of PHA production. PHA productivity (0.0697 g L−1 h−1), PHA concentration (8.37 g L−1), and PHA content (39.52%) were the highest when ARF was fed every 3 h for 61 h. This study proved that condensed corn solubles can be potentially used as a growth medium to boost PHA production by R. eutropha thus reducing the overall cost of biopolymer production. PMID:23118512

PHB granules are attached to the nucleoid via PhaM in Ralstonia eutropha.

PubMed

Wahl, Andreas; Schuth, Nora; Pfeiffer, Daniel; Nussberger, Stephan; Jendrossek, Dieter

2012-11-16

Poly(3-hydroxybutyrate) (PHB) granules are important storage compounds of carbon and energy in many prokaryotes which allow survival of the cells in the absence of suitable carbon sources. Formation and subcellular localization of PHB granules was previously assumed to occur randomly in the cytoplasm of PHB accumulating bacteria. However, contradictionary results on subcellular localization of PHB granules in Ralstonia eutropha were published, recently. Here, we provide evidence by transmission electron microscopy that PHB granules are localized in close contact to the nucleoid region in R. eutropha during growth on nutrient broth. Binding of PHB granules to the nucleoid is mediated by PhaM, a PHB granule associated protein with phasin-like properties that is also able to bind to DNA and to phasin PhaP5. Over-expression of PhaM resulted in formation of many small PHB granules that were always attached to the nucleoid region. In contrast, PHB granules of ∆phaM strains became very large and distribution of granules to daughter cells was impaired. Association of PHB granules to the nucleoid region was prevented by over-expression of PhaP5 and clusters of several PHB granules were mainly localized near the cell poles. Subcellular localization of PHB granules is controlled in R. eutropha and depends on the presence and concentrations of at least two PHB granule associated proteins, PhaM and PhaP5.

Polyhydroxyalkanoates production with Ralstonia eutropha from low quality waste animal fats.

PubMed

Riedel, Sebastian L; Jahns, Stefan; Koenig, Steven; Bock, Martina C E; Brigham, Christopher J; Bader, Johannes; Stahl, Ulf

2015-11-20

Polyhydroxyalkanoates (PHAs) are biodegradable and biocompatible polyesters considered as alternatives to petroleum-based plastics. Ralstonia eutropha is a model organism for PHA production. Utilizing industrially rendered waste animal fats as inexpensive carbon feedstocks for PHA production is demonstrated here. An emulsification strategy, without any mechanical or chemical pre-treatment, was developed to increase the bioavailability of solid, poorly-consumable fats. Wild type R. eutropha strain H16 produced 79-82% (w/w) polyhydroxybutyrate (PHB) per cell dry weight (CDW) when cultivated on various fats. A productivity of 0.3g PHB/(L × h) with a total PHB production of 24 g/L was achieved using tallow as carbon source. Using a recombinant strain of R. eutropha that produces poly(hydroxybutyrate-co-hydroxyhexanoate) [P(HB-co-HHx)], 49-72% (w/w) of PHA per CDW with a HHx content of 16-27 mol% were produced in shaking flask experiments. The recombinant strain was grown on waste animal fat of the lowest quality available at lab fermenter scale, resulting in 45 g/L CDW with 60% (w/w) PHA per CDW and a productivity of 0.4 g PHA/(L × h). The final HHx content of the polymer was 19 mol%. The use of low quality waste animal fats as an inexpensive carbon feedstock exhibits a high potential to accelerate the commercialization of PHAs. Copyright © 2015 Elsevier B.V. All rights reserved.

Localization of Poly(3-Hydroxybutyrate) (PHB) Granule-Associated Proteins during PHB Granule Formation and Identification of Two New Phasins, PhaP6 and PhaP7, in Ralstonia eutropha H16

PubMed Central

Pfeiffer, Daniel

2012-01-01

Poly(3-hydroxybutyrate) (PHB) granules are covered by a surface layer consisting of mainly phasins and other PHB granule-associated proteins (PGAPs). Phasins are small amphiphilic proteins that determine the number and size of accumulated PHB granules. Five phasin proteins (PhaP1 to PhaP5) are known for Ralstonia eutropha. In this study, we identified three additional potential phasin genes (H16_B1988, H16_B2296, and H16_B2326) by inspection of the R. eutropha genome for sequences with “phasin 2 motifs.” To determine whether the corresponding proteins represent true PGAPs, fusions with eYFP (enhanced yellow fluorescent protein) were constructed. Similar fusions of eYFP with PhaP1 to PhaP5 as well as fusions with PHB synthase (PhaC1), an inactive PhaC1 variant (PhaC1-C319A), and PhaC2 were also made. All fusions were investigated in wild-type and PHB-negative backgrounds. Colocalization with PHB granules was found for all PhaC variants and for PhaP1 to PhaP5. Additionally, eYFP fusions with H16_B1988 and H16_B2326 colocalized with PHB. Fusions of H16_B2296 with eYFP, however, did not colocalize with PHB granules but did colocalize with the nucleoid region. Notably, all fusions (except H16_B2296) were soluble in a ΔphaC1 strain. These data confirm that H16_B1988 and H16_B2326 but not H16_B2296 encode true PGAPs, for which we propose the designation PhaP6 (H16_B1988) and PhaP7 (H16_B2326). When localization of phasins was investigated at different stages of PHB accumulation, fusions of PhaP6 and PhaP7 were soluble in the first 3 h under PHB-permissive conditions, although PHB granules appeared after 10 min. At later time points, the fusions colocalized with PHB. Remarkably, PHB granules of strains expressing eYFP fusions with PhaP5, PhaP6, or PhaP7 localized predominantly near the cell poles or in the area of future septum formation. This phenomenon was not observed for the other PGAPs (PhaP1 to PhaP4, PhaC1, PhaC1-C319A, and PhaC2) and indicated that some phasins can have additional functions. A chromosomal deletion of phaP6 or phaP7 had no visible effect on formation of PHB granules. PMID:22923598

Localization of poly(3-hydroxybutyrate) (PHB) granule-associated proteins during PHB granule formation and identification of two new phasins, PhaP6 and PhaP7, in Ralstonia eutropha H16.

PubMed

Pfeiffer, Daniel; Jendrossek, Dieter

2012-11-01

Poly(3-hydroxybutyrate) (PHB) granules are covered by a surface layer consisting of mainly phasins and other PHB granule-associated proteins (PGAPs). Phasins are small amphiphilic proteins that determine the number and size of accumulated PHB granules. Five phasin proteins (PhaP1 to PhaP5) are known for Ralstonia eutropha. In this study, we identified three additional potential phasin genes (H16_B1988, H16_B2296, and H16_B2326) by inspection of the R. eutropha genome for sequences with "phasin 2 motifs." To determine whether the corresponding proteins represent true PGAPs, fusions with eYFP (enhanced yellow fluorescent protein) were constructed. Similar fusions of eYFP with PhaP1 to PhaP5 as well as fusions with PHB synthase (PhaC1), an inactive PhaC1 variant (PhaC1-C319A), and PhaC2 were also made. All fusions were investigated in wild-type and PHB-negative backgrounds. Colocalization with PHB granules was found for all PhaC variants and for PhaP1 to PhaP5. Additionally, eYFP fusions with H16_B1988 and H16_B2326 colocalized with PHB. Fusions of H16_B2296 with eYFP, however, did not colocalize with PHB granules but did colocalize with the nucleoid region. Notably, all fusions (except H16_B2296) were soluble in a ΔphaC1 strain. These data confirm that H16_B1988 and H16_B2326 but not H16_B2296 encode true PGAPs, for which we propose the designation PhaP6 (H16_B1988) and PhaP7 (H16_B2326). When localization of phasins was investigated at different stages of PHB accumulation, fusions of PhaP6 and PhaP7 were soluble in the first 3 h under PHB-permissive conditions, although PHB granules appeared after 10 min. At later time points, the fusions colocalized with PHB. Remarkably, PHB granules of strains expressing eYFP fusions with PhaP5, PhaP6, or PhaP7 localized predominantly near the cell poles or in the area of future septum formation. This phenomenon was not observed for the other PGAPs (PhaP1 to PhaP4, PhaC1, PhaC1-C319A, and PhaC2) and indicated that some phasins can have additional functions. A chromosomal deletion of phaP6 or phaP7 had no visible effect on formation of PHB granules.

Efficient biological conversion of carbon monoxide (CO) to carbon dioxide (CO2) and for utilization in bioplastic production by Ralstonia eutropha through the display of an enzyme complex on the cell surface.

PubMed

Hyeon, Jeong Eun; Kim, Seung Wook; Park, Chulhwan; Han, Sung Ok

2015-06-25

An enzyme complex for biological conversion of CO to CO2 was anchored on the cell surface of the CO2-utilizing Ralstonia eutropha and successfully resulted in a 3.3-fold increase in conversion efficiency. These results suggest that this complexed system may be a promising strategy for CO2 utilization as a biological tool for the production of bioplastics.

Production of branched-chain alcohols by recombinant Ralstonia eutropha in fed-batch cultivation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Fei, Q; Brigham, CJ; Lu, JN

Branched-chain alcohols are considered promising green energy sources due to their compatibility with existing infrastructure and their high energy density. We utilized a strain of Ralstonia eutropha capable of producing branched-chain alcohols and examined its production in flask cultures. In order to increase isobutanol and 3-methyl-1-butanol (isoamyl alcohol) productivity in the engineered strain, batch, fed-batch, and two-stage fed-batch cultures were carried out in this work. The effects of nitrogen source concentration on branched-chain alcohol production were investigated under four different initial concentrations in fermenters. A maximum 380 g m(-3) of branched-chain alcohol production was observed with 2 kg m(-3) initialmore » NH4Cl concentration in batch cultures. A pH-stat control strategy was utilized to investigate the optimum carbon source amount fed during fed-batch cultures for higher cell density. In cultures of R. eutropha strains that did not produce polyhydroxyalkanoate or branched-chain alcohols, a maximum cell dry weight of 36 kg m(-3) was observed using a fed-batch strategy, when 10 kg m(-3) carbon source was fed into culture medium. Finally, a total branched-chain alcohol titer of 790 g m(-3), the highest branched-chain alcohol yield of 0.03 g g(-1), and the maximum branched-chain alcohol productivity of 8.23 g m(-3) h(-1) were obtained from the engineered strain Re2410/pJL26 in a two-stage fed-batch culture system with pH-stat control. Isobutanol made up over 95% (mass fraction) of the total branched-chain alcohols titer produced in this study. (C) 2013 Published by Elsevier Ltd.« less

Degradation of formaldehyde at high concentrations by phenol-adapted Ralstonia eutropha closely related to pink-pigmented facultative methylotrophs.

PubMed

Habibi, Alireza; Vahabzadeh, Farzaneh

2013-01-01

The ability of the phenol-adapted Ralstonia eutropha to utilize formaldehyde (FD) as the sole source of carbon and energy was studied. Adaptation to FD was accomplished by substituting FD for glucose in a stepwise manner. The bacterium in the liquid test culture could tolerate concentrations of FD up to 900 mg L(-1). Degradation of FD was complete in 528 h at 30°C with shaking at 150 rpm (r = 1.67 mg L(-1) h(-1)), q = 0.035 g(FD) g(cell) (-1) h(-1). Substrate inhibition kinetics (Haldane and Luong equations) are used to describe the experimental data. At non-inhibitory concentrations of FD, the Monod equation was used. According to the Luong model, the values of the maximum specific growth rate (μ(max)), half-saturation coefficient (k(S)), the maximum allowable formaldehyde concentration (S(m)), and the shape factor (n) were 0.117 h(-1), 47.6 mg L(-1), 900 mg L(-1), and 2.2, respectively. The growth response of the test bacterium to consecutive FD feedings was examined, and the FD-adapted R. eutropha cells were able to degrade 1000 mg L(-1) FD in 150 h through 4 cycles of FD feeds. During FD degradation, formic acid metabolite was formed. Assimilation of FD, methanol, formic acid, and oxalate by the test bacterium was accompanied by the formation of a pink pigment. The carotenoid nature of the cellular pigment has been confirmed and the test bacterium appeared to be closely related to pink-pigmented facultative methylotrophs (PPFM). The extent of harm to soil exposed to biotreated wastewaters containing FD may be moderated due to the association between methylotrophic/oxalotrophic bacteria and plants.

Chemoselective Nitro Group Reduction and Reductive Dechlorination Initiate Degradation of 2-Chloro-5-Nitrophenol by Ralstonia eutropha JMP134

PubMed Central

Schenzle, Andreas; Lenke, Hiltrud; Spain, Jim C.; Knackmuss, Hans-Joachim

1999-01-01

Ralstonia eutropha JMP134 utilizes 2-chloro-5-nitrophenol as a sole source of nitrogen, carbon, and energy. The initial steps for degradation of 2-chloro-5-nitrophenol are analogous to those of 3-nitrophenol degradation in R. eutropha JMP134. 2-Chloro-5-nitrophenol is initially reduced to 2-chloro-5-hydroxylaminophenol, which is subject to an enzymatic Bamberger rearrangement yielding 2-amino-5-chlorohydroquinone. The chlorine of 2-amino-5-chlorohydroquinone is removed by a reductive mechanism, and aminohydroquinone is formed. 2-Chloro-5-nitrophenol and 3-nitrophenol induce the expression of 3-nitrophenol nitroreductase, of 3-hydroxylaminophenol mutase, and of the dechlorinating activity. 3-Nitrophenol nitroreductase catalyzes chemoselective reduction of aromatic nitro groups to hydroxylamino groups in the presence of NADPH. 3-Nitrophenol nitroreductase is active with a variety of mono-, di-, and trinitroaromatic compounds, demonstrating a relaxed substrate specificity of the enzyme. Nitrosobenzene serves as a substrate for the enzyme and is converted faster than nitrobenzene. PMID:10347008

Whole-Genome Microarray and Gene Deletion Studies Reveal Regulation of the Polyhydroxyalkanoate Production Cycle by the Stringent Response in Ralstonia eutropha H16

DOE Office of Scientific and Technical Information (OSTI.GOV)

Brigham, CJ; Speth, DR; Rha, C

Poly(3-hydroxybutyrate) (PHB) production and mobilization in Ralstonia eutropha are well studied, but in only a few instances has PHB production been explored in relation to other cellular processes. We examined the global gene expression of wild-type R. eutropha throughout the PHB cycle: growth on fructose, PHB production using fructose following ammonium depletion, and PHB utilization in the absence of exogenous carbon after ammonium was resupplied. Our results confirm or lend support to previously reported results regarding the expression of PHB-related genes and enzymes. Additionally, genes for many different cellular processes, such as DNA replication, cell division, and translation, are selectivelymore » repressed during PHB production. In contrast, the expression levels of genes under the control of the alternative sigma factor sigma(54) increase sharply during PHB production and are repressed again during PHB utilization. Global gene regulation during PHB production is strongly reminiscent of the gene expression pattern observed during the stringent response in other species. Furthermore, a ppGpp synthase deletion mutant did not show an accumulation of PHB, and the chemical induction of the stringent response with DL-norvaline caused an increased accumulation of PHB in the presence of ammonium. These results indicate that the stringent response is required for PHB accumulation in R. eutropha, helping to elucidate a thus-far-unknown physiological basis for this process.« less

PHB granules are attached to the nucleoid via PhaM in Ralstonia eutropha

PubMed Central

2012-01-01

Background Poly(3-hydroxybutyrate) (PHB) granules are important storage compounds of carbon and energy in many prokaryotes which allow survival of the cells in the absence of suitable carbon sources. Formation and subcellular localization of PHB granules was previously assumed to occur randomly in the cytoplasm of PHB accumulating bacteria. However, contradictionary results on subcellular localization of PHB granules in Ralstonia eutropha were published, recently. Results Here, we provide evidence by transmission electron microscopy that PHB granules are localized in close contact to the nucleoid region in R. eutropha during growth on nutrient broth. Binding of PHB granules to the nucleoid is mediated by PhaM, a PHB granule associated protein with phasin-like properties that is also able to bind to DNA and to phasin PhaP5. Over-expression of PhaM resulted in formation of many small PHB granules that were always attached to the nucleoid region. In contrast, PHB granules of ∆phaM strains became very large and distribution of granules to daughter cells was impaired. Association of PHB granules to the nucleoid region was prevented by over-expression of PhaP5 and clusters of several PHB granules were mainly localized near the cell poles. Conclusion Subcellular localization of PHB granules is controlled in R. eutropha and depends on the presence and concentrations of at least two PHB granule associated proteins, PhaM and PhaP5. PMID:23157596

The multifunctional isopropyl alcohol dehydrogenase of Phytomonas sp. could be the result of a horizontal gene transfer from a bacterium to the trypanosomatid lineage.

PubMed

Molinas, Sara M; Altabe, Silvia G; Opperdoes, Fred R; Rider, Mark H; Michels, Paul A M; Uttaro, Antonio D

2003-09-19

Isopropyl alcohol dehydrogenase (iPDH) is a dimeric mitochondrial alcohol dehydrogenase (ADH), so far detected within the Trypanosomatidae only in the genus Phytomonas. The cloning, sequencing, and heterologous expression of the two gene alleles of the enzyme revealed that it is a zinc-dependent medium-chain ADH. Both polypeptides have 361 amino acids. A mitochondrial targeting sequence was identified. The mature proteins each have 348 amino acids and a calculated molecular mass of 37 kDa. They differ only in one amino acid, which can explain the three isoenzymes and their respective isoelectric points previously found. A phylogenetic analysis locates iPDH within a cluster with fermentative ADHs from bacteria, sharing 74% similarity and 60% identity with Ralstonia eutropha ADH. The characterization of the two bacterially expressed Phytomonas enzymes and the comparison of their kinetic properties with those of the wild-type iPDH and of the R. eutropha ADH strongly support the idea of a horizontal gene transfer event from a bacterium to a trypanosomatid to explain the origin of the iPDH in Phytomonas. Phytomonas iPDH and R. eutropha ADH are able to use a wide range of substrates with similar Km values such as primary and secondary alcohols, diols, and aldehydes, as well as ketones such as acetone, diacetyl, and acetoin. We speculate that, as for R. eutropha ADH, Phytomonas iPDH acts as a safety valve for the release of excess reducing power.

Generation of poly-β-hydroxybutyrate from acetate in higher plants: Detection of acetoacetyl CoA reductase- and PHB synthase- activities in rice.

PubMed

Tsuda, Hirohisa; Shiraki, Mari; Inoue, Eri; Saito, Terumi

2016-08-20

It has been reported that Poly-β-hydroxybutyrate (PHB) is generated from acetate in the rice root. However, no information is available about the biosynthetic pathway of PHB from acetate in plant cells. In the bacterium Ralstonia eutropha H16 (R. eutropha), PHB is synthesized from acetyl CoA by the consecutive reaction of three enzymes: β-ketothiolase (EC: 2.3.1.9), acetoacetyl CoA reductase (EC: 1.1.1.36) and PHB synthase (EC: 2.3.1.-). Thus, in this study, we examined whether the above three enzymatic activities were also detected in rice seedlings. The results clearly showed that the activities of the above three enzymes were all detected in rice. In particular, the PHB synthase activity was detected specifically in the sonicated particulate fractions (2000g 10min precipitate (ppt) and the 8000g 30min ppt) of rice roots and leaves. In addition to these enzyme activities, several new experimental results were obtained on PHB synthesis in higher plants: (a) (14)C-PHB generated from 2-(14)C-acetate was mainly localized in the 2000g 10min ppt and the 8000g 30min ppt of rice root. (b) Addition of acetate (0.1-10mM) to culture medium of rice seedlings did not increase the content of PHB in the rice root or leaf. (c) In addition to C3 plants, PHB was generated from acetate in a C4 plant (corn) and in a CAM plant (Bryophyllum pinnatum). d) Washing with ethylenediaminetetraacetic acid (EDTA) strongly suggested that the PHB synthesized from acetate was of plant origin and was not bacterial contamination. Copyright © 2016 Elsevier GmbH. All rights reserved.

Comparative and Functional Genomics of Rhodococcus opacus PD630 for Biofuels Development

PubMed Central

Holder, Jason W.; Ulrich, Jil C.; DeBono, Anthony C.; Godfrey, Paul A.; Desjardins, Christopher A.; Zucker, Jeremy; Zeng, Qiandong; Leach, Alex L. B.; Ghiviriga, Ion; Dancel, Christine; Abeel, Thomas; Gevers, Dirk; Kodira, Chinnappa D.; Desany, Brian; Affourtit, Jason P.; Birren, Bruce W.; Sinskey, Anthony J.

2011-01-01

The Actinomycetales bacteria Rhodococcus opacus PD630 and Rhodococcus jostii RHA1 bioconvert a diverse range of organic substrates through lipid biosynthesis into large quantities of energy-rich triacylglycerols (TAGs). To describe the genetic basis of the Rhodococcus oleaginous metabolism, we sequenced and performed comparative analysis of the 9.27 Mb R. opacus PD630 genome. Metabolic-reconstruction assigned 2017 enzymatic reactions to the 8632 R. opacus PD630 genes we identified. Of these, 261 genes were implicated in the R. opacus PD630 TAGs cycle by metabolic reconstruction and gene family analysis. Rhodococcus synthesizes uncommon straight-chain odd-carbon fatty acids in high abundance and stores them as TAGs. We have identified these to be pentadecanoic, heptadecanoic, and cis-heptadecenoic acids. To identify bioconversion pathways, we screened R. opacus PD630, R. jostii RHA1, Ralstonia eutropha H16, and C. glutamicum 13032 for growth on 190 compounds. The results of the catabolic screen, phylogenetic analysis of the TAGs cycle enzymes, and metabolic product characterizations were integrated into a working model of prokaryotic oleaginy. PMID:21931557

Characterization of poly-3-hydroxybutyrate (PHB) produced from Ralstonia eutropha using an alkali-pretreated biomass feedstock.

PubMed

Saratale, Ganesh D; Oh, Min-Kyu

2015-09-01

Alkaline pretreatment using NaOH, KOH, or NaOCl has been applied to various types of waste biomass to enhance enzymatic digestibility. Pretreatment (2% NaOH, 121 °C, 30 min) of rice paddy straw (PS) resulted in a maximum yield of 703 mg of reducing sugar per gram of PS with 84.19% hydrolysis yield after a two-step enzymatic hydrolysis process. Ralstonia eutropha ATCC 17699 was tested for its ability to synthesize poly-3-hydroxybutyrate (PHB) using PS hydrolysates as its sole carbon source. It is noteworthy that dry cell weight, polyhydroxyalkanoate (PHA) accumulation and PHB yield with the use of laboratory-grade sugars were similar to those achieved with PS-derived sugars. Under optimized conditions, we observed maximal PHA accumulation (75.45%) and PHB production (11.42 g/L) within 48 h of fermentation. After PHB recovery, the physicochemical properties of PHB were determined by various analytical techniques, showed the results were consistent with the characteristics of a standard polymer of PHB. Thus, the PS hydrolysate proved to be an excellent cheap carbon substrate for PHB production. Copyright © 2015 Elsevier B.V. All rights reserved.

Cloning and Expression of a Ralstonia eutropha HF39 Gene Mediating Indigo Formation in Escherichia coli

PubMed Central

Drewlo, Sascha; Brämer, Christian O.; Madkour, Mohamed; Mayer, Frank; Steinbüchel, Alexander

2001-01-01

On complex medium Escherichia coli strains carrying hybrid plasmid pBEC/EE:11.0, pSKBEC/BE:9.0, pSKBEC/PP:3.3, or pSKBEC/PP:2.4 harboring genomic DNA of Ralstonia eutropha HF39 produced a blue pigment characterized as indigo by several chemical and spectroscopic methods. A 1,251-bp open reading frame (bec) was cloned and sequenced. The deduced amino acid sequence of bec showed only weak similarities to short-chain acyl-coenzyme A dehydrogenases, and the gene product catalyzed formation of indoxyl, a reactive preliminary stage for production of indigo. PMID:11282658

Versatile plasmid-based expression systems for Gram-negative bacteria--General essentials exemplified with the bacterium Ralstonia eutropha H16.

PubMed

Gruber, Steffen; Schwab, Helmut; Koefinger, Petra

2015-12-25

The Gram-negative bacterium Escherichia coli is currently the most efficient and widely used prokaryotic host for recombinant protein and metabolite production. However, due to some limitations and to various interesting features of other Gram-negative bacteria efficient vector systems applicable to a broad range are desired. Basic building blocks for plasmid-based vectors include besides the need for a suitable selection marker in the first line a proper replication and maintenance system. In addition to these basic requirements, further elements are needed for Gram-negative bacteria beyond E. coli, such as Pseudomonas pudita, Ralstonia eutropha, Burkholderia glumae or Acinetobacter sp.. Established building blocks have to be adapted and new building blocks providing the desired functions need to be identified and exploited. This minireview addresses so far described and used genetic elements for broad host range replication, efficient plasmid maintenance, and conjugative plasmid transfer as well as expression elements and protein secretion signals. The industrially important bacterium R. eutropha H16 was chosen as a model organism to provide specific data on the effectivity and utility of building blocks based on such genetic elements. Copyright © 2015 Elsevier B.V. All rights reserved.

Impact of Ralstonia eutropha's poly(3-Hydroxybutyrate) (PHB) Depolymerases and Phasins on PHB storage in recombinant Escherichia coli.

PubMed

Eggers, Jessica; Steinbüchel, Alexander

2014-12-01

The model organism for polyhydroxybutyrate (PHB) biosynthesis, Ralstonia eutropha H16, possesses multiple isoenzymes of granules coating phasins as well as of PHB depolymerases, which degrade accumulated PHB under conditions of carbon limitation. In this study, recombinant Escherichia coli BL21(DE3) strains were used to study the impact of selected PHB depolymerases of R. eutropha H16 on the growth behavior and on the amount of accumulated PHB in the absence or presence of phasins. For this purpose, 20 recombinant E. coli BL21(DE3) strains were constructed, which harbored a plasmid carrying the phaCAB operon from R. eutropha H16 to ensure PHB synthesis and a second plasmid carrying different combinations of the genes encoding a phasin and a PHB depolymerase from R. eutropha H16. It is shown in this study that the growth behavior of the respective recombinant E. coli strains was barely affected by the overexpression of the phasin and PHB depolymerase genes. However, the impact on the PHB contents was significantly greater. The strains expressing the genes of the PHB depolymerases PhaZ1, PhaZ2, PhaZ3, and PhaZ7 showed 35% to 94% lower PHB contents after 30 h of cultivation than the control strain. The strain harboring phaZ7 reached by far the lowest content of accumulated PHB (only 2.0% [wt/wt] PHB of cell dry weight). Furthermore, coexpression of phasins in addition to the PHB depolymerases influenced the amount of PHB stored in cells of the respective strains. It was shown that the phasins PhaP1, PhaP2, and PhaP4 are not substitutable without an impact on the amount of stored PHB. In particular, the phasins PhaP2 and PhaP4 seemed to limit the degradation of PHB by the PHB depolymerases PhaZ2, PhaZ3, and PhaZ7, whereas almost no influence of the different phasins was observed if phaZ1 was coexpressed. This study represents an extensive analysis of the impact of PHB depolymerases and phasins on PHB accumulation and provides a deeper insight into the complex interplay of these enzymes. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Impact of Ralstonia eutropha's Poly(3-Hydroxybutyrate) (PHB) Depolymerases and Phasins on PHB Storage in Recombinant Escherichia coli

PubMed Central

Eggers, Jessica

2014-01-01

The model organism for polyhydroxybutyrate (PHB) biosynthesis, Ralstonia eutropha H16, possesses multiple isoenzymes of granules coating phasins as well as of PHB depolymerases, which degrade accumulated PHB under conditions of carbon limitation. In this study, recombinant Escherichia coli BL21(DE3) strains were used to study the impact of selected PHB depolymerases of R. eutropha H16 on the growth behavior and on the amount of accumulated PHB in the absence or presence of phasins. For this purpose, 20 recombinant E. coli BL21(DE3) strains were constructed, which harbored a plasmid carrying the phaCAB operon from R. eutropha H16 to ensure PHB synthesis and a second plasmid carrying different combinations of the genes encoding a phasin and a PHB depolymerase from R. eutropha H16. It is shown in this study that the growth behavior of the respective recombinant E. coli strains was barely affected by the overexpression of the phasin and PHB depolymerase genes. However, the impact on the PHB contents was significantly greater. The strains expressing the genes of the PHB depolymerases PhaZ1, PhaZ2, PhaZ3, and PhaZ7 showed 35% to 94% lower PHB contents after 30 h of cultivation than the control strain. The strain harboring phaZ7 reached by far the lowest content of accumulated PHB (only 2.0% [wt/wt] PHB of cell dry weight). Furthermore, coexpression of phasins in addition to the PHB depolymerases influenced the amount of PHB stored in cells of the respective strains. It was shown that the phasins PhaP1, PhaP2, and PhaP4 are not substitutable without an impact on the amount of stored PHB. In particular, the phasins PhaP2 and PhaP4 seemed to limit the degradation of PHB by the PHB depolymerases PhaZ2, PhaZ3, and PhaZ7, whereas almost no influence of the different phasins was observed if phaZ1 was coexpressed. This study represents an extensive analysis of the impact of PHB depolymerases and phasins on PHB accumulation and provides a deeper insight into the complex interplay of these enzymes. PMID:25281380

Phasin Proteins Activate Aeromonas caviae Polyhydroxyalkanoate (PHA) Synthase but Not Ralstonia eutropha PHA Synthase

PubMed Central

Ushimaru, Kazunori; Motoda, Yoko; Numata, Keiji

2014-01-01

In this study, we performed in vitro and in vivo activity assays of polyhydroxyalkanoate (PHA) synthases (PhaCs) in the presence of phasin proteins (PhaPs), which revealed that PhaPs are activators of PhaC derived from Aeromonas caviae (PhaCAc). In in vitro assays, among the three PhaCs tested, PhaCAc was significantly activated when PhaPs were added at the beginning of polymerization (prepolymerization PhaCAc), whereas the prepolymerization PhaCRe (derived from Ralstonia eutropha) and PhaCDa (Delftia acidovorans) showed reduced activity with PhaPs. The PhaP-activated PhaCAc showed a slight shift of substrate preference toward 3-hydroxyhexanoyl-CoA (C6). PhaPAc also activated PhaCAc when it was added during polymerization (polymer-elongating PhaCAc), while this effect was not observed for PhaCRe. In an in vivo assay using Escherichia coli TOP10 as the host strain, the effect of PhaPAc expression on PHA synthesis by PhaCAc or PhaCRe was examined. As PhaPAc expression increased, PHA production was increased by up to 2.3-fold in the PhaCAc-expressing strain, whereas it was slightly increased in the PhaCRe-expressing strain. Taken together, this study provides evidence that PhaPs function as activators for PhaCAc both in vitro and in vivo but do not activate PhaCRe. This activating effect may be attributed to the new role of PhaPs in the polymerization reaction by PhaCAc. PMID:24584238

Characterization of Acrylamidase isolated from a newly isolated acrylamide-utilizing bacterium, Ralstonia eutropha AUM-01.

PubMed

Cha, Minseok; Chambliss, Glenn H

2011-02-01

A mesophilic bacterium capable of utilizing acrylamide was isolated, AUM-01, from soil collected from leaf litter at Picnic Point on the UW-Madison campus. In minimal medium with acrylamide as the sole carbon and nitrogen source, a batch culture of AUM-01 completely converted 28.0 mM acrylamide to acrylic acid in 8 h and reached a cell density of 0.3 (A₆₀₀)). Afterward all the acrylic acid was degraded by 20 h with the cell density increasing to 1.9 (A₆₀₀). The acrylamide-utilizing bacterium was identified as Ralstonia eutropha based on morphological observations, the BiOLOG GN2 MicroPlate™ identification system for Gram-negative bacteria, and additional physiological tests. An acrylamidase that hydrolyzes acrylamide to acrylic acid was purified from the strain AUM-01. The molecular weight of the enzyme from AUM-01 was determined to be 38 kDa by SDS-PAGE. The enzyme had pH and temperature optima of 6.3 and 55°C, and the influence of different metals and amino acids on the ability of the purified protein to transform acrylamide to acrylic acid was evaluated. The enzyme from AUM-01 was totally inhibited by ZnSO₄ and AgNO₃.

Spectroscopic insights into the oxygen-tolerant membrane-associated [NiFe] hydrogenase of Ralstonia eutropha H16.

PubMed

Saggu, Miguel; Zebger, Ingo; Ludwig, Marcus; Lenz, Oliver; Friedrich, Bärbel; Hildebrandt, Peter; Lendzian, Friedhelm

2009-06-12

This study provides the first spectroscopic characterization of the membrane-bound oxygen-tolerant [NiFe] hydrogenase (MBH) from Ralstonia eutropha H16 in its natural environment, the cytoplasmic membrane. The H2-converting MBH is composed of a large subunit, harboring the [NiFe] active site, and a small subunit, capable in coordinating one [3Fe4S] and two [4Fe4S] clusters. The hydrogenase dimer is electronically connected to a membrane-integral cytochrome b. EPR and Fourier transform infrared spectroscopy revealed a strong similarity of the MBH active site with known [NiFe] centers from strictly anaerobic hydrogenases. Most redox states characteristic for anaerobic [NiFe] hydrogenases were identified except for one remarkable difference. The formation of the oxygen-inhibited Niu-A state was never observed. Furthermore, EPR data showed the presence of an additional paramagnetic center at high redox potential (+290 mV), which couples magnetically to the [3Fe4S] center and indicates a structural and/or redox modification at or near the proximal [4Fe4S] cluster. Additionally, significant differences regarding the magnetic coupling between the Nia-C state and [4Fe4S] clusters were observed in the reduced form of the MBH. The spectroscopic properties are discussed with regard to the unusual oxygen tolerance of this hydrogenase and in comparison with those of the solubilized, dimeric form of the MBH.

Catalytic properties of the isolated diaphorase fragment of the NAD-reducing [NiFe]-hydrogenase from Ralstonia eutropha.

PubMed

Lauterbach, Lars; Idris, Zulkifli; Vincent, Kylie A; Lenz, Oliver

2011-01-01

The NAD+-reducing soluble hydrogenase (SH) from Ralstonia eutropha H16 catalyzes the H₂-driven reduction of NAD+, as well as reverse electron transfer from NADH to H+, in the presence of O₂. It comprises six subunits, HoxHYFUI₂, and incorporates a [NiFe] H+/H₂ cycling catalytic centre, two non-covalently bound flavin mononucleotide (FMN) groups and an iron-sulfur cluster relay for electron transfer. This study provides the first characterization of the diaphorase sub-complex made up of HoxF and HoxU. Sequence comparisons with the closely related peripheral subunits of Complex I in combination with UV/Vis spectroscopy and the quantification of the metal and FMN content revealed that HoxFU accommodates a [2Fe2S] cluster, FMN and a series of [4Fe4S] clusters. Protein film electrochemistry (PFE) experiments show clear electrocatalytic activity for both NAD+ reduction and NADH oxidation with minimal overpotential relative to the potential of the NAD+/NADH couple. Michaelis-Menten constants of 56 µM and 197 µM were determined for NADH and NAD+, respectively. Catalysis in both directions is product inhibited with K(I) values of around 0.2 mM. In PFE experiments, the electrocatalytic current was unaffected by O₂, however in aerobic solution assays, a moderate superoxide production rate of 54 nmol per mg of protein was observed, meaning that the formation of reactive oxygen species (ROS) observed for the native SH can be attributed mainly to HoxFU. The results are discussed in terms of their implications for aerobic functioning of the SH and possible control mechanism for the direction of catalysis.

Engineering Ralstonia eutropha for Production of Isobutanol (IBT) Motor Fuel from Carbon Dioxide, Hydrogen, and Oxygen Project Final Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sinskey, Anthony J.; Worden, Robert Mark; Brigham, Christopher

This research project is a collaboration between the Sinskey laboratory at MIT and the Worden laboratory at Michigan State University. The goal of the project is to produce Isobutanol (IBT), a branched-chain alcohol that can serve as a drop-in transportation fuel, through the engineered microbial biosynthesis of Carbon Dioxide, Hydrogen, and Oxygen using a novel bioreactor. This final technical report presents the findings of both the biological engineering work at MIT that extended the native branched-chain amino acid pathway of the wild type Ralstonia eutropha H16 to perform this biosynthesis, as well as the unique design, modeling, and construction ofmore » a bioreactor for incompatible gasses at Michigan State that enabled the operational testing of the complete system. This 105 page technical report summarizing the three years of research includes 72 figures and 11 tables of findings. Ralstonia eutropha (also known as Cupriavidus necator) is a Gram-negative, facultatively chemolithoautotrophic bacteria. It has been the principle organism used for the study of polyhydroxybutyrate (PHB) polymer biosynthesis. The wild-type Ralstonia eutropha H16 produces PHB as an intracellular carbon storage material while under nutrient stress in the presence of excess carbon. Under this stress, it can accumulate approximately 80 % of its cell dry weight (CDW) as this intracellular polymer. With the restoration of the required nutrients, the cells are then able to catabolize this polymer. If extracted from the cell, this PHB polymer can be processed into biodegradable and biocompatible plastics, however for this research, it is the efficient metabolic pathway channeling the captured carbon that is of interest. R. eutropha is further unique in that it contains two carbon-fixation Calvin–Benson–Bassham cycle operons, two oxygen-tolerant hydrogenases, and several formate dehydrogenases. It has also been much studied for its ability in the presence of oxygen, to fix carbon dioxide into complex cellular molecules using the energy from hydrogen. In this research project, engineered strains of R. eutropha redirected the excess carbon from PHB storage into the production of isobutanol and 3-methyl-1-butanol (branched-chain higher alcohols). These branched-chain higher alcohols can be used directly as substitutes for fossil-based fuels and are seen as alternative biofuels to ethanol and biodiesel. Importantly, these alcohols have approximately 98 % of the energy content of gasoline, 17 % higher than the current gasoline additive ethanol, without impacting corn market production for feed or food. Unlike ethanol, these branched-chain alcohols have low vapor pressure, hygroscopicity, and water solubility, which make them readily compatible with the existing pipelines, gasoline pumps, and engines in our transportation infrastructure. While the use of alternative energies from solar, wind, geothermal, and hydroelectric has spread for stationary power applications, these energy sources cannot be effectively or efficiently employed in current or future transportation systems. With the ongoing concerns of fossil fuel availability and price stability over the long term, alternative biofuels like branched-chain higher alcohols hold promise as a suitable transportation fuel in the future. We showed in our research that various mutant strains of R. eutropha with isobutyraldehyde dehydrogenase activity, in combination with the overexpression of plasmid-borne, native branched-chain amino acid biosynthesis pathway genes and the overexpression of heterologous ketoisovalerate decarboxylase gene, would produce isobutanol and 3-methyl-1-butanol when initiated during nitrogen or phosphorus limitation. Early on, we isolated one mutant R. eutropha strain which produced over 180 mg/L branched-chain alcohols in flask culture while being more tolerant of isobutanol toxicity. After the targeted elimination of genes encoding several potential carbon sinks (ilvE, bkdAB, and aceE), the production titer of the improved to 270 mg/L isobutanol and 40 mg/L 3-methyl-1-butanol. Semicontinuous flask cultivation supplied the cells with sufficient nutrients while minimizing the toxicity caused by isobutanol. Under this cultivation, the R. eutropha mutant grew and produced more than 14 g/L branched-chain alcohols over the duration of 50 days. These results demonstrate that R. eutropha carbon flux can be redirected from PHB to branched-chain alcohols and that engineered R. eutropha can be cultivated over prolonged periods of time for product biosynthesis. While this bioengineering work was being done at the Sinskey laboratory at MIT, the researchers at the Worden laboratory at Michigan State were working on the design and construction of the required specialty bioreactor for incompatible gasses (BIG) that would allow the safe feeding of microbes on Carbon Dioxide, Hydrogen, and Oxygen without explosive results. The early design and assembly work in year 1 incorporated a novel microbubble generator to maximize the bioavailability of gasses within the system comprised of small scale hollow fiber reactors. The early success of the microbubble generator eliminated the need to investigate potentially toxic surfactants within the system. For operational control, the system design incorporated a Opto22-based control network. The researchers also selected the specific hollow fiber material suitable for the bioreactor application. A variety of commercially available hollow fiber membranes were compared with regard to their pore sizes, cell affinity, and potential interference to cell viability assays. The selected membrane with its spongy layer was then tested for diffusivity of O2 and CO2. The instrumented system was then fully assembled for experimentally measuring the heterotrophic growth rate of immobilized R. eutropha cells. The requisite procedures for inoculation, measurement, and cleaning were established enabling the system performance to be validated under controlled laboratory conditions. In year 2, the researchers completed the Opto22 based cross-platform control network, and the system’s communications across the Sartorius fermentation system and Bruker gas chromatograph was established via open platform communications (OPC) protocol. Using the revised system, measurements were taken of the R.eutropha cell growth rate and substrate mass transfer rate in the hollow fiber membrane. Several IBT recovery strategies were explored and resin adsorption was determined to be optimal solution for lab scale operations. The adsorption capacity of the resin column was then measured and IBT desorption using methanol has been demonstrated. With the growing body of experimental data in hand, mathematical models were constructed to demonstrate and map the cellular kinetics, mass transfer of heterotrophic and autotrophic substrates in the hollow fiber, and the adsorption process in the resin column. A structured kinetic model was constructed to describe the competition between cell mass generation and IBT production. The reactor was then scaled up from single fiber to a membrane area of 180 cm2 and then further to 1 ft2. In Year 3 of the research, the IBT mass transfer across the membrane was characterized within the system with experiments to empirically measure the IBT diffusion coefficient in the BIG spongy layer. Using the refined mathematical models, the researchers are now able to explain the experimental observations and predict bioreactor performance under a wide range of experimental conditions. The Big system is able to demonstrate continuous controlled operations with the integrated IBT recovery system. Both heterotrophic and autotrophic production have been shown during continuous operation with heterotrophic and autotrophic stages. Performance of BIG system has been measured during continuous run with alternating heterotrophic growth on fructose and autotrophic product formation on H2, CO2, and O2. Volumetric productivities of IBT at 325 mg/(L day) and of 3M1B at 50 mg/(L day) were achieved, which were comparable to that achieved under heterotrophic conditions. Using the mathematical model, researchers are able to predict system performance for scaled-up BIG system. The apparent diffusion coefficient of IBT in the spongy layer of XM-50 hollow fiber membranes has been measured at various lumen liquid flow rates. The experiment is simulated in COMSOL to validate the results. The constructed COMSOL model is able to simulate BIG system performance in both batch and continuous mode. Mathematical simulations of the system performance have been run to identify the most crucial operational conditions, identifying the rate-limiting factors in autotrophic production of IBT, and quantitating the rate of IBT catabolism. Investigations of the productivity of the production system have suggested and the modeling of the system has revealed a particular sensitivity to the catabolism of the produced IBT by the engineered R. eutropha. Experiments have been designed and executed to quantify the IBT catabolism of R. eutropha, which open up possibilities for further system improvements through future, targeted bioengineering of the strain. Finally, the researchers at Michigan State performed an economic analysis of the system, based on the collective results, and their findings are presented in full within the report.« less

The H(2) sensor of Ralstonia eutropha: biochemical and spectroscopic analysis of mutant proteins modified at a conserved glutamine residue close to the [NiFe] active site.

PubMed

Buhrke, Thorsten; Brecht, Marc; Lubitz, Wolfgang; Friedrich, Bärbel

2002-09-01

[NiFe] hydrogenases contain a highly conserved histidine residue close to the [NiFe] active site which is altered by a glutamine residue in the H(2)-sensing [NiFe] hydrogenases. In this study, we exchanged the respective glutamine residue of the H(2) sensor (RH) of Ralstonia eutropha, Q67 of the RH large subunit HoxC, by histidine, asparagine and glutamate. The replacement by histidine and asparagine resulted in slightly unstable RH proteins which were hardly affected in their regulatory and enzymatic properties. The exchange to glutamate led to a completely unstable RH protein. The purified wild-type RH and the mutant protein with the Gln/His exchange were analysed by continuous-wave and pulsed electron paramagnetic resonance (EPR) techniques. We observed a coupling of a nitrogen nucleus with the [NiFe] active site for the mutant protein which was absent in the spectrum of the wild-type RH. A combination of theoretical calculations with the experimental data provided an explanation for the observed coupling. It is shown that the coupling is due to the formation of a weak hydrogen bond between the protonated N(epsilon) nucleus of the histidine with the sulfur of a conserved cysteine residue which coordinates the metal atoms of the [NiFe] active site as a bridging ligand. The effect of this hydrogen bond on the local structure of the [NiFe] active site is discussed.

H2 conversion in the presence of O2 as performed by the membrane-bound [NiFe]-hydrogenase of Ralstonia eutropha.

PubMed

Lenz, Oliver; Ludwig, Marcus; Schubert, Torsten; Bürstel, Ingmar; Ganskow, Stefanie; Goris, Tobias; Schwarze, Alexander; Friedrich, Bärbel

2010-04-26

[NiFe]-hydrogenases catalyze the oxidation of H(2) to protons and electrons. This reversible reaction is based on a complex interplay of metal cofactors including the Ni-Fe active site and several [Fe-S] clusters. H(2) catalysis of most [NiFe]-hydrogenases is sensitive to dioxygen. However, some bacteria contain hydrogenases that activate H(2) even in the presence of O(2). There is now compelling evidence that O(2) affects hydrogenase on three levels: 1) H(2) catalysis, 2) hydrogenase maturation, and 3) H(2)-mediated signal transduction. Herein, we summarize the genetic, biochemical, electrochemical, and spectroscopic properties related to the O(2) tolerance of hydrogenases resident in the facultative chemolithoautotroph Ralstonia eutropha H16. A focus is given to the membrane-bound [NiFe]-hydogenase, which currently represents the best-characterized member of O(2)-tolerant hydrogenases.

Improved apparatus for continuous culture of hydrogen-fixing bacteria

NASA Technical Reports Server (NTRS)

Foster, J. F.; Litchfield, J. H.

1970-01-01

Improved apparatus permits the continuous culture of Hydrogenomonas eutropha. System incorporates three essential subsystems - /1/ environmentally isolated culture vessel, /2/ analytical system with appropriate sensors and readout devices, /3/ control system with feedback responses to each analytical measurement.

Hydrogen-driven asymmetric reduction of hydroxyacetone to (R)-1,2-propanediol by Ralstonia eutropha transformant expressing alcohol dehydrogenase from Kluyveromyces lactis.

PubMed

Oda, Takahiro; Oda, Koji; Yamamoto, Hiroaki; Matsuyama, Akinobu; Ishii, Masaharu; Igarashi, Yasuo; Nishihara, Hirofumi

2013-01-10

Conversion of industrial processes to more nature-friendly modes is a crucial subject for achieving sustainable development. Utilization of hydrogen-oxidation reactions by hydrogenase as a driving force of bioprocess reaction can be an environmentally ideal method because the reaction creates no pollutants. We expressed NAD-dependent alcohol dehydrogenase from Kluyveromyces lactis in a hydrogen-oxidizing bacterium: Ralstonia eutropha. This is the first report of hydrogen-driven in vivo coupling reaction of the alcohol dehydrogenase and indigenous soluble NAD-reducing hydrogenase. Asymmetric reduction of hydroxyacetone to (R)-1,2-propanediol, which is a commercial building block for antibacterial agents, was performed using the transformant as the microbial cell catalyst. The two enzymes coupled in vitro in vials without a marked decrease of reactivity during the 20 hr reaction because of the hydrogenase reaction, which generates no by-product that affects enzymes. Alcohol dehydrogenase was expressed functionally in R. eutropha in an activity level equivalent to that of indigenous NAD-reducing hydrogenase under the hydrogenase promoter. The hydrogen-driven in vivo coupling reaction proceeded only by the transformant cell without exogenous addition of a cofactor. The decrease of reaction velocity at higher concentration of hydroxyacetone was markedly reduced by application of an in vivo coupling system. Production of (R)-1,2-propanediol (99.8% e.e.) reached 67.7 g/l in 76 hr with almost a constant rate using a jar fermenter. The reaction velocity under 10% PH2 was almost equivalent to that under 100% hydrogen, indicating the availability of crude hydrogen gas from various sources. The in vivo coupling system enabled cell-recycling as catalysts. Asymmetric reduction of hydroxyacetone by a coupling reaction of the two enzymes continued in both in vitro and in vivo systems in the presence of hydrogen. The in vivo reaction system using R. eutropha transformant expressing heterologous alcohol dehydrogenase showed advantages for practical usage relative to the in vitro coupling system. The results suggest a hopeful perspective of the hydrogen-driven bioprocess as an environmentally outstanding method to achieve industrial green innovation. Hydrogen-oxidizing bacteria can be useful hosts for the development of hydrogen-driven microbial cell factories.

Nutrition Knowledge in the U.S. Navy

DTIC Science & Technology

1990-12-31

rAD-A232 085]Y( NUTRITION KNOWLEDGE IN THE U.S. NAVY L. K. TRENT kI DT|C-A91" AfELECTEj •,bMAR 1 ~ REPORT No. 90-27 Approved for public release...BETHESDA, MARYLAND 91 3 11 034 NUTRITION KNOWLEDGE IN THE U. S. NAVY Linda Kelly Trent Health Services Research Department Naval Health Research Center P.O...implicating diet in the etiology of disease continues to mount, national organizations have launched nutrition education campaigns intended to guide the

Novel 2,4-Dichlorophenoxyacetic Acid Degradation Genes from Oligotrophic Bradyrhizobium sp. Strain HW13 Isolated from a Pristine Environment

PubMed Central

Kitagawa, Wataru; Takami, Sachiko; Miyauchi, Keisuke; Masai, Eiji; Kamagata, Yoichi; Tiedje, James M.; Fukuda, Masao

2002-01-01

The tfd genes of Ralstonia eutropha JMP134 are the only well-characterized set of genes responsible for 2,4-dichlorophenoxyacetic acid (2,4-D) degradation among 2,4-D-degrading bacteria. A new family of 2,4-D degradation genes, cadRABKC, was cloned and characterized from Bradyrhizobium sp. strain HW13, a strain that was isolated from a buried Hawaiian soil that has never experienced anthropogenic chemicals. The cadR gene was inferred to encode an AraC/XylS type of transcriptional regulator from its deduced amino acid sequence. The cadABC genes were predicted to encode 2,4-D oxygenase subunits from their deduced amino acid sequences that showed 46, 44, and 37% identities with the TftA and TftB subunits of 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) oxygenase of Burkholderia cepacia AC1100 and with a putative ferredoxin, ThcC, of Rhodococcus erythropolis NI86/21, respectively. They are thoroughly different from the 2,4-D dioxygenase gene, tfdA, of R. eutropha JMP134. The cadK gene was presumed to encode a 2,4-D transport protein from its deduced amino acid sequence that showed 60% identity with the 2,4-D transporter, TfdK, of strain JMP134. Sinorhizobium meliloti Rm1021 cells containing cadRABKC transformed several phenoxyacetic acids, including 2,4-D and 2,4,5-T, to corresponding phenol derivatives. Frameshift mutations indicated that each of the cadRABC genes was essential for 2,4-D conversion in strain Rm1021 but that cadK was not. Five 2,4-D degraders, including Bradyrhizobium and Sphingomonas strains, were found to have cadA gene homologs, suggesting that these 2,4-D degraders share 2,4-D degradation genes similar to those of strain HW13 cadABC. PMID:11751829

Comparative proteome analysis reveals four novel polyhydroxybutyrate (PHB) granule-associated proteins in Ralstonia eutropha H16.

PubMed

Sznajder, Anna; Pfeiffer, Daniel; Jendrossek, Dieter

2015-03-01

Identification of proteins that were present in a polyhydroxybutyrate (PHB) granule fraction isolated from Ralstonia eutropha but absent in the soluble, membrane, and membrane-associated fractions revealed the presence of only 12 polypeptides with PHB-specific locations plus 4 previously known PHB-associated proteins with multiple locations. None of the previously postulated PHB depolymerase isoenzymes (PhaZa2 to PhaZa5, PhaZd1, and PhaZd2) and none of the two known 3-hydroxybutyrate oligomer hydrolases (PhaZb and PhaZc) were significantly present in isolated PHB granules. Four polypeptides were found that had not yet been identified in PHB granules. Three of the novel proteins are putative α/β-hydrolases, and two of those (A0671 and B1632) have a PHB synthase/depolymerase signature. The third novel protein (A0225) is a patatin-like phospholipase, a type of enzyme that has not been described for PHB granules of any PHB-accumulating species. No function has been ascribed to the fourth protein (A2001), but its encoding gene forms an operon with phaB2 (acetoacetyl-coenzyme A [CoA] reductase) and phaC2 (PHB synthase), and this is in line with a putative function in PHB metabolism. The localization of the four new proteins at the PHB granule surface was confirmed in vivo by fluorescence microscopy of constructed fusion proteins with enhanced yellow fluorescent protein (eYFP). Deletion of A0671 and B1632 had a minor but detectable effect on the PHB mobilization ability in the stationary growth phase of nutrient broth (NB)-gluconate cells, confirming the functional involvement of both proteins in PHB metabolism. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Comparative Proteome Analysis Reveals Four Novel Polyhydroxybutyrate (PHB) Granule-Associated Proteins in Ralstonia eutropha H16

PubMed Central

Sznajder, Anna; Pfeiffer, Daniel

2014-01-01

Identification of proteins that were present in a polyhydroxybutyrate (PHB) granule fraction isolated from Ralstonia eutropha but absent in the soluble, membrane, and membrane-associated fractions revealed the presence of only 12 polypeptides with PHB-specific locations plus 4 previously known PHB-associated proteins with multiple locations. None of the previously postulated PHB depolymerase isoenzymes (PhaZa2 to PhaZa5, PhaZd1, and PhaZd2) and none of the two known 3-hydroxybutyrate oligomer hydrolases (PhaZb and PhaZc) were significantly present in isolated PHB granules. Four polypeptides were found that had not yet been identified in PHB granules. Three of the novel proteins are putative α/β-hydrolases, and two of those (A0671 and B1632) have a PHB synthase/depolymerase signature. The third novel protein (A0225) is a patatin-like phospholipase, a type of enzyme that has not been described for PHB granules of any PHB-accumulating species. No function has been ascribed to the fourth protein (A2001), but its encoding gene forms an operon with phaB2 (acetoacetyl-coenzyme A [CoA] reductase) and phaC2 (PHB synthase), and this is in line with a putative function in PHB metabolism. The localization of the four new proteins at the PHB granule surface was confirmed in vivo by fluorescence microscopy of constructed fusion proteins with enhanced yellow fluorescent protein (eYFP). Deletion of A0671 and B1632 had a minor but detectable effect on the PHB mobilization ability in the stationary growth phase of nutrient broth (NB)-gluconate cells, confirming the functional involvement of both proteins in PHB metabolism. PMID:25548058

Fixation of carbon dioxide by a hydrogen-oxidizing bacterium for value-added products.

PubMed

Yu, Jian

2018-06-09

With rapid technology progress and cost reduction, clean hydrogen from water electrolysis driven by renewable powers becomes a potential feedstock for CO 2 fixation by hydrogen-oxidizing bacteria. Cupriavidus necator (formally Ralstonia eutropha), a representative member of the lithoautotrophic prokaryotes, is a promising producer of polyhydroxyalkanoates and single cell proteins. This paper reviews the fundamental properties of the hydrogen-oxidizing bacterium, the metabolic activities under limitation of individual gases and nutrients, and the value-added products from CO 2 , including the products with large potential markets. Gas fermentation and bioreactor safety are discussed for achieving high cell density and high productivity of desired products under chemolithotrophic conditions. The review also updates the recent research activities in metabolic engineering of C. necator to produce novel metabolites from CO 2 .

Crystal structure and biochemical characterization of beta-keto thiolase B from polyhydroxyalkanoate-producing bacterium Ralstonia eutropha H16

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kim, Eun-Jung; Son, Hyeoncheol Francis; Kim, Sangwoo

Highlights: • We determined a crystal structure of β-keto thiolase from Ralstonia eutropha H16 (ReBktB). • Distinct substrate binding mode ReBktB was elucidated. • Enzymatic kinetic parameters of ReBktB were revealed. - Abstract: ReBktB is a β-keto thiolase from Ralstonia eutropha H16 that catalyzes condensation reactions between acetyl-CoA with acyl-CoA molecules that contains different numbers of carbon atoms, such as acetyl-CoA, propionyl-CoA, and butyryl-CoA, to produce valuable bioproducts, such as polyhydroxybutyrate, polyhydroxybutyrate-hydroxyvalerate, and hexanoate. We solved a crystal structure of ReBktB at 2.3 Å, and the overall structure has a similar fold to that of type II biosynthetic thiolases, suchmore » as PhbA from Zoogloea ramigera (ZrPhbA). The superposition of this structure with that of ZrPhbA complexed with CoA revealed the residues that comprise the catalytic and substrate binding sites of ReBktB. The catalytic site of ReBktB contains three conserved residues, Cys90, His350, and Cys380, which may function as a covalent nucleophile, a general base, and second nucleophile, respectively. For substrate binding, ReBktB stabilized the ADP moiety of CoA in a distinct way compared to ZrPhbA with His219, Arg221, and Asp228 residues, whereas the stabilization of β-mercaptoethyamine and pantothenic acid moieties of CoA was quite similar between these two enzymes. Kinetic study of ReBktB revealed that K{sub m}, V{sub max}, and K{sub cat} values of 11.58 μM, 1.5 μmol/min, and 102.18 s{sup −1}, respectively, and the catalytic and substrate binding sites of ReBktB were further confirmed by site-directed mutagenesis experiments.« less

The Development and Recognition of Homeland Security Law

DTIC Science & Technology

2007-03-01

Facilitation” (CTPAT)90; “ AML / CFT compliance91”; Bioshield92; two votes for Legislative and/or Government relations; “Implications for Intellectual...NAVAL POSTGRADUATE SCHOOL MONTEREY, CALIFORNIA THESIS Approved for public release; distribution is unlimited THE DEVELOPMENT AND...RECOGNITION OF HOMELAND SECURITY LAW by Michael C. McDaniel March 2007 Thesis Advisor: Chris Bellavita Second Reader

New XML-Based Files: Implications for Forensics

DTIC Science & Technology

2009-04-01

previously unknown social networks.4 We can use unique identi!ers that survived copying and pasting to show plagiarism . Unique identi!ers can also raise...graduated with honors and received the Admiral Grace Murray Hopper award for academic achievement. <a:"d id=’’{985AE863-DF53-4B19-9956-91DEFC2F01C1

3-Hydroxylaminophenol Mutase from Ralstonia eutropha JMP134 Catalyzes a Bamberger Rearrangement

PubMed Central

Schenzle, Andreas; Lenke, Hiltrud; Spain, Jim C.; Knackmuss, Hans-Joachim

1999-01-01

3-Hydroxylaminophenol mutase from Ralstonia eutropha JMP134 is involved in the degradative pathway of 3-nitrophenol, in which it catalyzes the conversion of 3-hydroxylaminophenol to aminohydroquinone. To show that the reaction was really catalyzed by a single enzyme without the release of intermediates, the corresponding protein was purified to apparent homogeneity from an extract of cells grown on 3-nitrophenol as the nitrogen source and succinate as the carbon and energy source. 3-Hydroxylaminophenol mutase appears to be a relatively hydrophobic but soluble and colorless protein consisting of a single 62-kDa polypeptide. The pI was determined to be at pH 4.5. In a database search, the NH2-terminal amino acid sequence of the undigested protein and of two internal sequences of 3-hydroxylaminophenol mutase were found to be most similar to those of glutamine synthetases from different species. Hydroxylaminobenzene, 4-hydroxylaminotoluene, and 2-chloro-5-hydroxylaminophenol, but not 4-hydroxylaminobenzoate, can also serve as substrates for the enzyme. The enzyme requires no oxygen or added cofactors for its reaction, which suggests an enzymatic mechanism analogous to the acid-catalyzed Bamberger rearrangement. PMID:10049374

Gas Consumption and Growth Rate of Hydrogenomonas eutropha in Continuous Culture

PubMed Central

Ammann, Elizabeth C. B.; Reed, Lawrence L.; Durichek, John E.

1968-01-01

The bacterium Hydrogenomonas eutropha is under consideration for use in a regenerative life-support system for manned space missions of long duration. A 4-liter continuous culture unit containing the organism was operated for a period of 272 days under autotrophic environmental conditions. The best steady-state run achieved with this unit was observed over a 22-day time interval after 181 days of operation. During this time, the culture consumed an average of 22.9 ± 2.0 ml of carbon dioxide per min, 38.1 ± 3.3 ml of oxygen per min, and 128.5 ± 10.6 ml of hydrogen per min. It required 18.7 ± 1.2 liters of fresh nutrient medium per 24 hr to maintain a constant, preestablished cell population of 1.65 g (dry weight) per liter. The ratio of consumption of carbon dioxide, oxygen, and hydrogen varied from 1:1.2:4.5 to 1:1.9:6.6, with an average of 1:1.7:5.7. Based on these values, approximately 60 liters of the culture would be necessary to balance the gas exchange of one man. Images Fig. 1 PMID:4385748

Spectroscopic and Kinetic Properties of the Molybdenum-containing, NAD+-dependent Formate Dehydrogenase from Ralstonia eutropha*

PubMed Central

Niks, Dimitri; Duvvuru, Jayant; Escalona, Miguel; Hille, Russ

2016-01-01

We have examined the rapid reaction kinetics and spectroscopic properties of the molybdenum-containing, NAD+-dependent FdsABG formate dehydrogenase from Ralstonia eutropha. We confirm previous steady-state studies of the enzyme and extend its characterization to a rapid kinetic study of the reductive half-reaction (the reaction of formate with oxidized enzyme). We have also characterized the electron paramagnetic resonance signal of the molybdenum center in its MoV state and demonstrated the direct transfer of the substrate Cα hydrogen to the molybdenum center in the course of the reaction. Varying temperature, microwave power, and level of enzyme reduction, we are able to clearly identify the electron paramagnetic resonance signals for four of the iron/sulfur clusters of the enzyme and find suggestive evidence for two others; we observe a magnetic interaction between the molybdenum center and one of the iron/sulfur centers, permitting assignment of this signal to a specific iron/sulfur cluster in the enzyme. In light of recent advances in our understanding of the structure of the molybdenum center, we propose a reaction mechanism involving direct hydride transfer from formate to a molybdenum-sulfur group of the molybdenum center. PMID:26553877

Efficient solar-to-fuels production from a hybrid microbial-water-splitting catalyst system.

PubMed

Torella, Joseph P; Gagliardi, Christopher J; Chen, Janice S; Bediako, D Kwabena; Colón, Brendan; Way, Jeffery C; Silver, Pamela A; Nocera, Daniel G

2015-02-24

Photovoltaic cells have considerable potential to satisfy future renewable-energy needs, but efficient and scalable methods of storing the intermittent electricity they produce are required for the large-scale implementation of solar energy. Current solar-to-fuels storage cycles based on water splitting produce hydrogen and oxygen, which are attractive fuels in principle but confront practical limitations from the current energy infrastructure that is based on liquid fuels. In this work, we report the development of a scalable, integrated bioelectrochemical system in which the bacterium Ralstonia eutropha is used to efficiently convert CO2, along with H2 and O2 produced from water splitting, into biomass and fusel alcohols. Water-splitting catalysis was performed using catalysts that are made of earth-abundant metals and enable low overpotential water splitting. In this integrated setup, equivalent solar-to-biomass yields of up to 3.2% of the thermodynamic maximum exceed that of most terrestrial plants. Moreover, engineering of R. eutropha enabled production of the fusel alcohol isopropanol at up to 216 mg/L, the highest bioelectrochemical fuel yield yet reported by >300%. This work demonstrates that catalysts of biotic and abiotic origin can be interfaced to achieve challenging chemical energy-to-fuels transformations.

Microbial production, ultrasound-assisted extraction and characterization of biopolymer polyhydroxybutyrate (PHB) from terrestrial (P. hysterophorus) and aquatic (E. crassipes) invasive weeds.

PubMed

Pradhan, Sushobhan; Borah, Arup Jyoti; Poddar, Maneesh Kumar; Dikshit, Pritam Kumar; Rohidas, Lilendar; Moholkar, Vijayanand S

2017-10-01

This study reports synthesis of biodegradable poly(3-hydroxybutyrate) (PHB) polymer from two invasive weeds, viz. P. hysterophorus and E. crassipes. The pentose and hexose-rich hydrolyzates obtained from acid pretreatment and enzymatic hydrolysis of two biomasses were separately fermented using Ralstonia eutropha MTCC 8320 sp. PHB was extracted using sonication and was characterized using FTIR, 1 H and 13 C NMR and XRD. PHB content of dry cell mass was 8.1-21.6% w/w, and the PHB yield was 6.85×10 -3 -36.41×10 -3 % w/w raw biomass. Thermal properties of PHB were determined by TGA, DTG and DSC analysis. PHB obtained from pentose-hydrolyzate had glass transition temperatures of 6°-9°C, while PHB from hexose-rich hydrolyzate had maximum thermal degradation temperatures of 370°-389°C. These thermal properties were comparable to the properties of commercial PHB. Probable causes leading to differences in thermal properties of pentose and hexose-derived PHB are: extent of crystallinity and presence of impurity in the polymer matrix. Copyright © 2017 Elsevier Ltd. All rights reserved.

Efficient solar-to-fuels production from a hybrid microbial–water-splitting catalyst system

PubMed Central

Torella, Joseph P.; Gagliardi, Christopher J.; Chen, Janice S.; Bediako, D. Kwabena; Colón, Brendan; Way, Jeffery C.; Silver, Pamela A.; Nocera, Daniel G.

2015-01-01

Photovoltaic cells have considerable potential to satisfy future renewable-energy needs, but efficient and scalable methods of storing the intermittent electricity they produce are required for the large-scale implementation of solar energy. Current solar-to-fuels storage cycles based on water splitting produce hydrogen and oxygen, which are attractive fuels in principle but confront practical limitations from the current energy infrastructure that is based on liquid fuels. In this work, we report the development of a scalable, integrated bioelectrochemical system in which the bacterium Ralstonia eutropha is used to efficiently convert CO2, along with H2 and O2 produced from water splitting, into biomass and fusel alcohols. Water-splitting catalysis was performed using catalysts that are made of earth-abundant metals and enable low overpotential water splitting. In this integrated setup, equivalent solar-to-biomass yields of up to 3.2% of the thermodynamic maximum exceed that of most terrestrial plants. Moreover, engineering of R. eutropha enabled production of the fusel alcohol isopropanol at up to 216 mg/L, the highest bioelectrochemical fuel yield yet reported by >300%. This work demonstrates that catalysts of biotic and abiotic origin can be interfaced to achieve challenging chemical energy-to-fuels transformations. PMID:25675518

Combination of N149S and D171G mutations in Aeromonas caviae polyhydroxyalkanoate synthase and impact on polyhydroxyalkanoate biosynthesis.

PubMed

Tsuge, Takeharu; Watanabe, Shinko; Shimada, Daisuke; Abe, Hideki; Doi, Yoshiharu; Taguchi, Seiichi

2007-12-01

Aeromonas caviae polyhydroxyalkanoate synthase (PhaC(Ac)) is an important biocatalyst for the synthesis of practically useful two-component polyhydroxyalkanoate copolymer, poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyhexanoate] [P(3HB-co-3HHx)]. In a previous study, two PhaC(Ac) mutants that have a single amino acid substitution of either asparagine 149 by serine (N149S) or aspartate 171 by glycine (D171G) were isolated as higher active enzymes by means of evolutionary engineering. In this study, the synergistic effects of N149S and D171G double mutation (NSDG) in PhaC(Ac) on polyhydroxyalkanoate biosynthesis were investigated in recombinant Ralstonia eutropha. The PhaC(Ac) NSDG mutant showed enhanced incorporation of longer 3-hydroxyalkanoate (3HA) units into the polyhydroxyalkanoate copolymer from octanoate (3HA fraction: 18.5 mol%) and soybean oil (5.4 mol%) as a carbon source. Besides, the NSDG mutant synthesized P(3HB) homopolymer with a very high molecular weight (M(w)=368 x 10(4)) when fructose was used as a carbon source. Thus, a combination of the beneficial mutations synergistically altered enzymatic properties, leading to synthesis of a polyhydroxyalkanoate copolymer with enhanced 3HA fraction and increased molecular weight.

Inhibitory effect of carbon dioxide on the fed-batch culture of Ralstonia eutropha: evaluation by CO2 pulse injection and autogenous CO2 methods.

PubMed

Shang, Longan; Jiang, Min; Ryu, Chul Hee; Chang, Ho Nam; Cho, Soon Haeng; Lee, Jong Won

2003-08-05

In order to see the effect of CO(2) inhibition resulting from the use of pure oxygen, we carried out a comparative fed-batch culture study of polyhydroxybutyric acid (PHB) production by Ralstonia eutropha using air and pure oxygen in 5-L, 30-L, and 300-L fermentors. The final PHB concentrations obtained with pure O(2) were 138.7 g/L in the 5-L fermentor and 131.3 g/L in the 30-L fermentor, which increased 2.9 and 6.2 times, respectively, as compared to those obtained with air. In the 300-L fermentor, the fed-batch culture with air yielded only 8.4 g/L PHB. However, the maximal CO(2) concentrations in the 5-L fermentor increased significantly from 4.1% (air) to 15.0% (pure O(2)), while it was only 1.6% in the 30-L fermentor with air, but reached 14.2% in the case of pure O(2). We used two different experimental methods for evaluating CO(2) inhibition: CO(2) pulse injection and autogenous CO(2) methods. A 10 or 22% (v/v) CO(2) pulse with a duration of 3 or 6 h was introduced in a pure-oxygen culture of R. eutropha to investigate how CO(2) affects the synthesis of biomass and PHB. CO(2) inhibited the cell growth and PHB synthesis significantly. The inhibitory effect became stronger with the increase of the CO(2) concentration and pulse duration. The new proposed autogenous CO(2) method makes it possible to place microbial cells under different CO(2) level environments by varying the gas flow rate. Introduction of O(2) gas at a low flow rate of 0.42 vvm resulted in an increase of CO(2) concentration to 30.2% in the exit gas. The final PHB of 97.2 g/L was obtained, which corresponded to 70% of the PHB production at 1.0 vvm O(2) flow rate. This new method measures the inhibitory effect of CO(2) produced autogenously by cells through the entire fermentation process and can avoid the overestimation of CO(2) inhibition without introducing artificial CO(2) into the fermentor. Copyright 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 312-320, 2003.

The auxiliary protein HypX provides oxygen tolerance to the soluble [NiFe]-hydrogenase of ralstonia eutropha H16 by way of a cyanide ligand to nickel.

PubMed

Bleijlevens, Boris; Buhrke, Thorsten; van der Linden, Eddy; Friedrich, Bärbel; Albracht, Simon P J

2004-11-05

The hypX gene of the facultative lithoautotrophic bacterium Ralstonia eutropha is part of a cassette of accessory genes (the hyp cluster) required for the proper assembly of the active site of the [NiFe]-hydrogenases in the bacterium. A deletion of the hypX gene led to a severe growth retardation under lithoautotrophic conditions with 5 or 15% oxygen, when the growth was dependent on the activity of the soluble NAD+ -reducing hydrogenase. The enzymatic and infrared spectral properties of the soluble hydrogenase purified from a HypX-negative strain were compared with those from an enzyme purified from a HypX-positive strain. In activity assays under anaerobic conditions both enzyme preparations behaved the same. Under aerobic conditions, however, the mutant enzyme became irreversibly inactivated during H2 oxidation with NAD+ or benzyl viologen as the electron acceptor. Infrared spectra and chemical determination of cyanide showed that one of the four cyanide groups in the wild-type enzyme was missing in the mutant enzyme. The data are consistent with the proposal that the HypX protein is specifically involved in the biosynthetic pathway that delivers the nickel-bound cyanide. The data support the proposal that this cyanide is crucial for the enzyme to function under aerobic conditions.

Ralstonia eutropha as a biocatalyst for desulfurization of dibenzothiophene.

PubMed

Dejaloud, Azita; Vahabzadeh, Farzaneh; Habibi, Alireza

2017-07-01

The potential of Ralstonia eutropha as a biocatalyst for desulfurization of dibenzothiophene (DBT) was studied in growing and resting cell conditions. The results of both conditions showed that sulfur was removed from DBT which accompanied by the formation of 2-hydroxybiphenyl (2-HBP). In growing cell experiments, glucose was used as an energy supplying substrate in initial concentrations of 55 mM (energy-limited) and 111 mM (energy-sufficient). The growing cell behaviors were quantitatively described using the logistic equation and maintenance concept. The results indicated that 2-HBP production was higher for the energy-sufficient cultures, while the values of the specific growth rate and the maintenance coefficient for these media were lower than those of the energy-limited cultures. Additionally, the kinetic studies showed that the half-saturation constant for the energy-limited cultures was 2 times higher than the energy-sufficient ones where the inhibition constant (0.08 mM) and the maximum specific DBT desulfurization rate (0.002 mmol g cell -1  h -1 ) were almost constant. By defining desulfurizing capacity (D DBT ) including both the biomass concentration and time to reach a particular percentage of DBT conversion, the best condition for desulfurizing cell was determined at 23% g cell L -1  h -1 which corresponded with the resting cells that were harvested at the mid-exponential growth phase.

Genome-wide sequencing and an open reading frame analysis of dichlorodiphenyltrichloroethane (DDT) susceptible (91-C) and resistant (91-R) Drosophila melanogaster laboratory populations

USDA-ARS?s Scientific Manuscript database

The Drosophila melanogaster 91-R and 91-C strains are of common origin, however, 91-R has been intensely selected for dichlorodiphenyltrichloroethane (DDT) resistance over six decades while 91-C has been maintained as the non-selected control strain. These fly strains represent a unique genetic res...

To Be or Not To Be a Poly(3-Hydroxybutyrate) (PHB) Depolymerase: PhaZd1 (PhaZ6) and PhaZd2 (PhaZ7) of Ralstonia eutropha, Highly Active PHB Depolymerases with No Detectable Role in Mobilization of Accumulated PHB

PubMed Central

Sznajder, Anna

2014-01-01

The putative physiological functions of two related intracellular poly(3-hydroxybutyrate) (PHB) depolymerases, PhaZd1 and PhaZd2, of Ralstonia eutropha H16 were investigated. Purified PhaZd1 and PhaZd2 were active with native PHB granules in vitro. Partial removal of the proteinaceous surface layer of native PHB granules by trypsin treatment or the use of PHB granules isolated from ΔphaP1 or ΔphaP1-phaP5 mutant strains resulted in increased specific PHB depolymerase activity, especially for PhaZd2. Constitutive expression of PhaZd1 or PhaZd2 reduced or even prevented the accumulation of PHB under PHB-permissive conditions in vivo. Expression of translational fusions of enhanced yellow fluorescent protein (EYFP) with PhaZd1 and PhaZd2 in which the active-site serines (S190 and Ser193) were replaced with alanine resulted in the colocalization of only PhaZd1 fusions with PHB granules. C-terminal fusions of inactive PhaZd2(S193A) with EYFP revealed the presence of spindle-like structures, and no colocalization with PHB granules was observed. Chromosomal deletion of phaZd1, phaZd2, or both depolymerase genes had no significant effect on PHB accumulation and mobilization during growth in nutrient broth (NB) or NB-gluconate medium. Moreover, neither proteome analysis of purified native PHB granules nor lacZ fusion studies gave any indication that PhaZd1 or PhaZd2 was detectably present in the PHB granule fraction or expressed at all during growth on NB-gluconate medium. In conclusion, PhaZd1 and PhaZd2 are two PHB depolymerases with a high capacity to degrade PHB when artificially expressed but are apparently not involved in PHB mobilization in the wild type. The true in vivo functions of PhaZd1 and PhaZd2 remain obscure. PMID:24907326

To be or not to be a poly(3-hydroxybutyrate) (PHB) depolymerase: PhaZd1 (PhaZ6) and PhaZd2 (PhaZ7) of Ralstonia eutropha, highly active PHB depolymerases with no detectable role in mobilization of accumulated PHB.

PubMed

Sznajder, Anna; Jendrossek, Dieter

2014-08-01

The putative physiological functions of two related intracellular poly(3-hydroxybutyrate) (PHB) depolymerases, PhaZd1 and PhaZd2, of Ralstonia eutropha H16 were investigated. Purified PhaZd1 and PhaZd2 were active with native PHB granules in vitro. Partial removal of the proteinaceous surface layer of native PHB granules by trypsin treatment or the use of PHB granules isolated from ΔphaP1 or ΔphaP1-phaP5 mutant strains resulted in increased specific PHB depolymerase activity, especially for PhaZd2. Constitutive expression of PhaZd1 or PhaZd2 reduced or even prevented the accumulation of PHB under PHB-permissive conditions in vivo. Expression of translational fusions of enhanced yellow fluorescent protein (EYFP) with PhaZd1 and PhaZd2 in which the active-site serines (S190 and Ser193) were replaced with alanine resulted in the colocalization of only PhaZd1 fusions with PHB granules. C-terminal fusions of inactive PhaZd2(S193A) with EYFP revealed the presence of spindle-like structures, and no colocalization with PHB granules was observed. Chromosomal deletion of phaZd1, phaZd2, or both depolymerase genes had no significant effect on PHB accumulation and mobilization during growth in nutrient broth (NB) or NB-gluconate medium. Moreover, neither proteome analysis of purified native PHB granules nor lacZ fusion studies gave any indication that PhaZd1 or PhaZd2 was detectably present in the PHB granule fraction or expressed at all during growth on NB-gluconate medium. In conclusion, PhaZd1 and PhaZd2 are two PHB depolymerases with a high capacity to degrade PHB when artificially expressed but are apparently not involved in PHB mobilization in the wild type. The true in vivo functions of PhaZd1 and PhaZd2 remain obscure. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Controlling microbial PHB synthesis via CRISPRi.

PubMed

Li, Dan; Lv, Li; Chen, Jin-Chun; Chen, Guo-Qiang

2017-07-01

Microbial polyhydroxyalkanoates (PHA) are a family of biopolyesters with properties similar to petroleum plastics such as polyethylene (PE) or polypropylene (PP). Polyhydroxybutyrate (PHB) is the most common PHA known so far. Clustered regularly interspaced short palindromic repeats interference (CRISPRi), a technology recently developed to control gene expression levels in eukaryotic and prokaryotic genomes, was employed to regulate PHB synthase activity influencing PHB synthesis. Recombinant Escherichia coli harboring an operon of three PHB synthesis genes phaCAB cloned from Ralstonia eutropha, was transformed with various single guided RNA (sgRNA with its guide sequence of 20-23 bases) able to bind to various locations of the PHB synthase PhaC, respectively. Depending on the binding location and the number of sgRNA on phaC, CRISPRi was able to control the phaC transcription and thus PhaC activity. It was found that PHB content, molecular weight, and polydispersity were approximately in direct and reverse proportion to the PhaC activity, respectively. The higher the PhaC activity, the more the intracellular PHB accumulation, yet the less the PHB molecular weights and the wider the polydispersity. This study allowed the PHB contents to be controlled in the ranges of 1.47-75.21% cell dry weights, molecular weights from 2 to 6 millions Dalton and polydispersity of 1.2 to 1.43 in 48 h shake flask studies. This result will be very important for future development of ultrahigh molecular weight PHA useful to meet high strength application requirements.

26 CFR 1.381(c)(9)-1 - Amortization of bond discount or premium.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 26 Internal Revenue 4 2010-04-01 2010-04-01 false Amortization of bond discount or premium. 1.381(c)(9)-1 Section 1.381(c)(9)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Insolvency Reorganizations § 1.381(c)(9)-1 Amortization of...

Job Language Performance Requirements for MOS 91C, Clinical Specialist, Reference Soldier’s Manual Dated 30 August 1977.

DTIC Science & Technology

1977-08-30

Administer a rectal suppository 081-91C-1240 Administer an injection (SC or IM) 081-91C-1241 Administer an intradermal injection 081-91C-1242 Administer a...in this language task: Shouting Radio communications Coded messages Spellings Conversation Requests wI-3 SPEAKING *TASK: Formulate and produce

Current Emergency Locator Transmitter (ELT) deficiencies and potential improvements utilizing TSO-C91a ELTs

NASA Technical Reports Server (NTRS)

Trudell, Bernard J.; Dreibelbis, Ryland R.

1990-01-01

An analysis was conducted of current ELT problems and potential improvements that could be made by employing the TSO-C91a ELTs to replace the current TSO-C91 ELTs. The scope of the study included the following: (1) validate the problems; (2) determine specific failure causes; (3) determine false alarm causes; (4) estimate improvements from TSO-C91a; (5) estimate benefits from replacement of the current ELTs; and (6) determine need and benefits for improved ELT inspection and maintenance. A detailed comparison between the two requirements documents (TSO-C91 and -91a) was made to assess improved performance of the ELT in each category of failure cause and each cause of false alarms. The comparison and analysis resulted in projecting a success of operation rate approximately 3 times the current rate and a reduction in false alarms to 0.25 of those generated by TSO-C91 ELTs. These improvements led to a projection of benefits of approximately 25 additional lives to be saved each year with TSO-C91a ELTs and an improved inspection and maintenance program.

Role of the HoxZ subunit in the electron transfer pathway of the membrane-bound [NiFe]-hydrogenase from Ralstonia eutropha immobilized on electrodes.

PubMed

Sezer, Murat; Frielingsdorf, Stefan; Millo, Diego; Heidary, Nina; Utesch, Tillman; Mroginski, Maria-Andrea; Friedrich, Bärbel; Hildebrandt, Peter; Zebger, Ingo; Weidinger, Inez M

2011-09-01

The role of the diheme cytochrome b (HoxZ) subunit in the electron transfer pathway of the membrane-bound [NiFe]-hydrogenase (MBH) heterotrimer from Ralstonia eutropha H16 has been investigated. The MBH in its native heterotrimeric state was immobilized on electrodes and subjected to spectroscopic and electrochemical analysis. Surface enhanced resonance Raman spectroscopy was used to monitor the redox and coordination state of the HoxZ heme cofactors while concomitant protein film voltammetric measurements gave insights into the catalytic response of the enzyme on the electrode. The entire MBH heterotrimer as well as its isolated HoxZ subunit were immobilized on silver electrodes coated with self-assembled monolayers of ω-functionalized alkylthiols, displaying the preservation of the native heme pocket structure and an electrical communication between HoxZ and the electrode. For the immobilized MBH heterotrimer, catalytic reduction of the HoxZ heme cofactors was observed upon H(2) addition. The catalytic currents of MBH with and without the HoxZ subunit were measured and compared with the heterogeneous electron transfer rates of the isolated HoxZ. On the basis of the spectroscopic and electrochemical results, we conclude that the HoxZ subunit under these artificial conditions is not primarily involved in the electron transfer to the electrode but plays a crucial role in stabilizing the enzyme on the electrode. © 2011 American Chemical Society

Site-directed Mutagenesis Reveals Regions Implicated in the Stability and Fiber Formation of Human λ3r Light Chains

DOE PAGES

Villalba, Miryam I.; Canul-Tec, Juan C.; Luna-Martínez, Oscar D.; ...

2014-12-11

Light chain amyloidosis (AL) is a disease that affects vital organs by the fibrillar aggregation of monoclonal light chains. λ3r germ line is significantly implicated in this disease. In this paper, we contrasted the thermodynamic stability and aggregation propensity of 3mJL2 (nonamyloidogenic) and 3rJL2 (amyloidogenic) λ3 germ lines. Because of an inherent limitation (extremely low expression), Cys at position 34 of the 3r germ line was replaced by Tyr reaching a good expression yield. A second substitution (W91A) was introduced in 3r to obtain a better template to incorporate additional mutations. Although the single mutant (C34Y) was not fibrillogenic, themore » second mutation located at CDR3 (W91A) induced fibrillogenesis. We propose, for the first time, that CDR3 (position 91) affects the stability and fiber formation of human λ3r light chains. Using the double mutant (3rJL2/YA) as template, other variants were constructed to evaluate the importance of those substitutions into the stability and aggregation propensity of λ3 light chains. A change in position 7 (P7D) boosted 3rJL2/YA fibrillogenic properties. Modification of position 48 (I48M) partially reverted 3rJL2/YA fibril aggregation. Finally, changes at positions 8 (P8S) or 40 (P40S) completely reverted fibril formation. These results confirm the influential roles of N-terminal region (positions 7 and 8) and the loop 40–60 (positions 40 and 48) on AL. X-ray crystallography revealed that the three-dimensional topology of the single and double λ3r mutants was not significantly altered. Finally, this mutagenic approach helped to identify key regions implicated in λ3 AL.« less

Identification and analysis of putative polyhydroxyalkanoate synthase (PhaC) in Pseudomonas fluorescens.

PubMed

Lim, Ju Hyoung; Rhie, Ho-Gun; Kim, Jeong Nam

2018-05-11

Pseudomonas fluorescens KLR101 was found to be capable of producing polyhydroxyalkanoate (PHA) using various sugars and fatty acids with carbon numbers ranging from 2 to 6. PHA granules mainly consisted of poly(3-hydroxybutyrate) homopolymer and/or poly(3-hydroxybutyrate- co -3-hydroxyvalerate) copolymer. Genomic DNA of P. fluorescens was fractionated and cloned into a lambda library, in which a 5.8-kb fragment hybridized to a heterologous phaC probe from Ralstonia eutropha was identified. In vivo expression in Klebsiella aerogenes KC2671 (pUMS), restriction mapping, Southern hybridization experiments, and sequencing data revealed that PHA biosynthesis by P. fluorescens relied upon a polypeptide encoded by a 1,683-bp non-operonal ORF, which was preceded by a possible -24/-12 promoter and highly similar to DNA sequences of a gene encoding PHA synthase in the genus Pseudomonas . In vivo expression of the putative PHA synthase gene ( phaC Pf ) in a recombinant Escherichia coli strain was investigated by using glucose and decanoate as substrates. E. coli ( phaC Pf + , pUMS) grown in medium containing glucose accumulated PHA granules mainly consisting of 3-hydroxybutyrate, whereas only a trace amount of 3-hydroxydecanoate was detected from E. coli fadR mutant ( phaC Pf + ) grown in medium containing decanoate. In vitro enzymatic assessment experiments showed that 3-hydroxybutyryl-CoA was efficiently used as a substrate of purified PhaC Pf , suggesting that the putative PHA synthase of P. fluorescens mainly utilizes short-chain-length PHA precursors as a substrate.

32 CFR Appendix C to Part 623 - Agreement for Loan of US Army Materiel (DA Form 4881-R)

Code of Federal Regulations, 2010 CFR

2010-07-01

... 32 National Defense 3 2010-07-01 2010-07-01 true Agreement for Loan of US Army Materiel (DA Form 4881-R) C Appendix C to Part 623 National Defense Department of Defense (Continued) DEPARTMENT OF THE... Loan of US Army Materiel (DA Form 4881-R) EC24OC91.013 EC24OC91.014 EC24OC91.015 EC24OC91.016 EC24OC91...

32 CFR Appendix C to Part 623 - Agreement for Loan of US Army Materiel (DA Form 4881-R)

Code of Federal Regulations, 2011 CFR

2011-07-01

... 32 National Defense 3 2011-07-01 2009-07-01 true Agreement for Loan of US Army Materiel (DA Form 4881-R) C Appendix C to Part 623 National Defense Department of Defense (Continued) DEPARTMENT OF THE... Loan of US Army Materiel (DA Form 4881-R) EC24OC91.013 EC24OC91.014 EC24OC91.015 EC24OC91.016 EC24OC91...

76 FR 68243 - Social Security Rulings, SSR 91-1c and SSR 66-18c; Rescission of Social Security Rulings (SSR) 66...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-11-03

..., Social Security Online, at http://www.socialsecurity.gov . SUPPLEMENTARY INFORMATION: SSRs make available... SOCIAL SECURITY ADMINISTRATION [Docket No. SSA-2011-0068] Social Security Rulings, SSR 91-1c and SSR 66-18c; Rescission of Social Security Rulings (SSR) 66-18c and SSR 91-1c AGENCY: Social Security...

Intramolecular and intermolecular N-H...C(5)H(5)(-) hydrogen bonding in magnesocene adducts of alkylamines. Implications for chemical vapor deposition using cyclopentadienyl source compounds.

PubMed

Xia, Aibing; Heeg, Mary Jane; Winter, Charles H

2002-09-25

Magnesocene adducts of alkylamines were prepared and characterized. Treatment of 3-amino-2,4-dimethylpentane, isopropylamine, tert-butylamine, benzylamine, or N-isopropylbenzylamine with magnesocene at ambient temperature in toluene afforded the amine adducts Cp2Mg(NH2CH(CH(CH3)2)2) (91%), Cp2Mg(NH2iPr) (80%), Cp2Mg(NH2tBu) (67%), Cp2Mg(NH2CH2Ph) (80%), and Cp2Mg(NH(CH(CH3)2)(CH2C6H5)) (91%). These adducts are stable at ambient temperature, and Cp2Mg(NH2CH(CH(CH3)2)2) can be sublimed at 60 degrees C/0.05 Torr without any evidence for reversion to magnesocene. The solid-state structure of Cp2Mg(NH2CH(CH(CH3)2)2) contains eta5- and eta2-cyclopentadienyl ligands, and the hydrogen atoms on the coordinated amine nitrogen atom participate in intramolecular and intermolecular hydrogen bonding to the eta2-cyclopentadienyl ligand. The observed hydrogen bonding is relevant to the path by which cyclopentadiene is eliminated from metal cyclopentadienyl CVD source compounds during film growth employing acidic element hydrides as co-reactants.

28 CFR 2.91 - Supervision responsibility.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 28 Judicial Administration 1 2011-07-01 2011-07-01 false Supervision responsibility. 2.91 Section 2.91 Judicial Administration DEPARTMENT OF JUSTICE PAROLE, RELEASE, SUPERVISION AND RECOMMITMENT OF....91 Supervision responsibility. (a) Pursuant to D.C. Code 24-133(c), the District of Columbia Court...

28 CFR 2.91 - Supervision responsibility.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 28 Judicial Administration 1 2010-07-01 2010-07-01 false Supervision responsibility. 2.91 Section 2.91 Judicial Administration DEPARTMENT OF JUSTICE PAROLE, RELEASE, SUPERVISION AND RECOMMITMENT OF....91 Supervision responsibility. (a) Pursuant to D.C. Code 24-133(c), the District of Columbia Court...

28 CFR 2.91 - Supervision responsibility.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 28 Judicial Administration 1 2012-07-01 2012-07-01 false Supervision responsibility. 2.91 Section 2.91 Judicial Administration DEPARTMENT OF JUSTICE PAROLE, RELEASE, SUPERVISION AND RECOMMITMENT OF....91 Supervision responsibility. (a) Pursuant to D.C. Code 24-133(c), the District of Columbia Court...

28 CFR 2.91 - Supervision responsibility.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 28 Judicial Administration 1 2013-07-01 2013-07-01 false Supervision responsibility. 2.91 Section 2.91 Judicial Administration DEPARTMENT OF JUSTICE PAROLE, RELEASE, SUPERVISION AND RECOMMITMENT OF....91 Supervision responsibility. (a) Pursuant to D.C. Code 24-133(c), the District of Columbia Court...

28 CFR 2.91 - Supervision responsibility.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 28 Judicial Administration 1 2014-07-01 2014-07-01 false Supervision responsibility. 2.91 Section 2.91 Judicial Administration DEPARTMENT OF JUSTICE PAROLE, RELEASE, SUPERVISION AND RECOMMITMENT OF....91 Supervision responsibility. (a) Pursuant to D.C. Code 24-133(c), the District of Columbia Court...

Polyhydroxyalkanoate (PHA) Granules Have no Phospholipids.

PubMed

Bresan, Stephanie; Sznajder, Anna; Hauf, Waldemar; Forchhammer, Karl; Pfeiffer, Daniel; Jendrossek, Dieter

2016-05-25

Polyhydroxybutyrate (PHB) granules, also designated as carbonosomes, are supra-molecular complexes in prokaryotes consisting of a PHB polymer core and a surface layer of structural and functional proteins. The presence of suspected phospholipids in the surface layer is based on in vitro data of isolated PHB granules and is often shown in cartoons of the PHB granule structure in reviews on PHB metabolism. However, the in vivo presence of a phospholipid layer has never been demonstrated. We addressed this topic by the expression of fusion proteins of DsRed2EC and other fluorescent proteins with the phospholipid-binding domain (LactC2) of lactadherin in three model organisms. The fusion proteins specifically localized at the cell membrane of Ralstonia eutropha but did not co-localize with PHB granules. The same result was obtained for Pseudomonas putida, a species that accumulates another type of polyhydroxyalkanoate (PHA) granules related to PHB. Notably, DsRed2EC-LactC2 expressed in Magnetospirillum gryphiswaldense was detected at the position of membrane-enclosed magnetosome chains and at the cytoplasmic membrane but not at PHB granules. In conclusion, the carbonosomes of representatives of α-proteobacteria, β-proteobacteria and γ-proteobacteria have no phospholipids in vivo and we postulate that the PHB/PHA granule surface layers in natural producers generally are free of phospholipids and consist of proteins only.

Bacterial Conversion of Hydroxylamino Aromatic Compounds by both Lyase and Mutase Enzymes Involves Intramolecular Transfer of Hydroxyl Groups

PubMed Central

Nadeau, Lloyd J.; He, Zhongqi; Spain, Jim C.

2003-01-01

Hydroxylamino aromatic compounds are converted to either the corresponding aminophenols or protocatechuate during the bacterial degradation of nitroaromatic compounds. The origin of the hydroxyl group of the products could be the substrate itself (intramolecular transfer mechanism) or the solvent water (intermolecular transfer mechanism). The conversion of hydroxylaminobenzene to 2-aminophenol catalyzed by a mutase from Pseudomonas pseudoalcaligenes JS45 proceeds by an intramolecular hydroxyl transfer. The conversions of hydroxylaminobenzene to 2- and 4-aminophenol by a mutase from Ralstonia eutropha JMP134 and to 4-hydroxylaminobenzoate to protocatechuate by a lyase from Comamonas acidovorans NBA-10 and Pseudomonas sp. strain 4NT were proposed, but not experimentally proved, to proceed by the intermolecular transfer mechanism. GC-MS analysis of the reaction products formed in H218O did not indicate any 18O-label incorporation during the conversion of hydroxylaminobenzene to 2- and 4-aminophenols catalyzed by the mutase from R. eutropha JMP134. During the conversion of 4-hydroxylaminobenzoate catalyzed by the hydroxylaminolyase from Pseudomonas sp. strain 4NT, only one of the two hydroxyl groups in the product, protocatechuate, was 18O labeled. The other hydroxyl group in the product must have come from the substrate. The mutase in strain JS45 converted 4-hydroxylaminobenzoate to 4-amino-3-hydroxybenzoate, and the lyase in Pseudomonas strain 4NT converted hydroxylaminobenzene to aniline and 2-aminophenol but not to catechol. The results indicate that all three types of enzyme-catalyzed rearrangements of hydroxylamino aromatic compounds proceed via intramolecular transfer of hydroxyl groups. PMID:12732549

An improved purification procedure for the soluble [NiFe]-hydrogenase of Ralstonia eutropha: new insights into its (in)stability and spectroscopic properties.

PubMed

van der Linden, Eddy; Burgdorf, Tanja; de Lacey, Antonio L; Buhrke, Thorsten; Scholte, Marcel; Fernandez, Victor M; Friedrich, Bärbel; Albracht, Simon P J

2006-03-01

Infrared (IR) spectra in combination with chemical analyses have recently shown that the active Ni-Fe site of the soluble NAD(+)-reducing [NiFe]-hydrogenase from Ralstonia eutropha contains four cyanide groups and one carbon monoxide as ligands. Experiments presented here confirm this result, but show that a variable percentage of enzyme molecules loses one or two of the cyanide ligands from the active site during routine purification. For this reason the redox conditions during the purification have been optimized yielding hexameric enzyme preparations (HoxFUYHI(2)) with aerobic specific H(2)-NAD(+) activities of 150-185 mumol/min/mg of protein (up to 200% of the highest activity previously reported in the literature). The preparations were highly homogeneous in terms of the active site composition and showed superior IR spectra. IR spectro-electrochemical studies were consistent with the hypothesis that only reoxidation of the reduced enzyme with dioxygen leads to the inactive state, where it is believed that a peroxide group is bound to nickel. Electron paramagnetic resonance experiments showed that the radical signal from the NADH-reduced enzyme derives from the semiquinone form of the flavin (FMN-a) in the hydrogenase module (HoxYH dimer), but not of the flavin (FMN-b) in the NADH-dehydrogenase module (HoxFU dimer). It is further demonstrated that the hexameric enzyme remains active in the presence of NADPH and air, whereas NADH and air lead to rapid destruction of enzyme activity. It is proposed that the presence of NADPH in cells keeps the enzyme in the active state.

Cloning of the Alcaligenes latus polyhydroxyalkanoate biosynthesis genes and use of these genes for enhanced production of Poly(3-hydroxybutyrate) in Escherichia coli.

PubMed

Choi, J I; Lee, S Y; Han, K

1998-12-01

Polyhydroxyalkanoates (PHAs) are microbial polyesters that can be used as completely biodegradable polymers, but the high production cost prevents their use in a wide range of applications. Recombinant Escherichia coli strains harboring the Ralstonia eutropha PHA biosynthesis genes have been reported to have several advantages as PHA producers compared with wild-type PHA-producing bacteria. However, the PHA productivity (amount of PHA produced per unit volume per unit time) obtained with these recombinant E. coli strains has been lower than that obtained with the wild-type bacterium Alcaligenes latus. To endow the potentially superior PHA biosynthetic machinery to E. coli, we cloned the PHA biosynthesis genes from A. latus. The three PHA biosynthesis genes formed an operon with the order PHA synthase, beta-ketothiolase, and reductase genes and were constitutively expressed from the natural promoter in E. coli. Recombinant E. coli strains harboring the A. latus PHA biosynthesis genes accumulated poly(3-hydroxybutyrate) (PHB), a model PHA product, more efficiently than those harboring the R. eutropha genes. With a pH-stat fed-batch culture of recombinant E. coli harboring a stable plasmid containing the A. latus PHA biosynthesis genes, final cell and PHB concentrations of 194.1 and 141.6 g/liter, respectively, were obtained, resulting in a high productivity of 4.63 g of PHB/liter/h. This improvement should allow recombinant E. coli to be used for the production of PHB with a high level of economic competitiveness.

Formation of a dinitrosyl iron complex by NorA, a nitric oxide-binding di-iron protein from Ralstonia eutropha H16.

PubMed

Strube, Katja; de Vries, Simon; Cramm, Rainer

2007-07-13

In Ralstonia eutropha H16, two genes, norA and norB, form a dicistronic operon that is controlled by the NO-responsive transcriptional regulator NorR. NorB has been identified as a membrane-bound NO reductase, but the physiological function of NorA is unknown. We found that, in a NorA deletion mutant, the promoter activity of the norAB operon was increased 3-fold, indicating that NorA attenuates activation of NorR. NorA shows limited sequence similarity to the oxygen carrier hemerythrin, which contains a di-iron center. Indeed, optical and EPR spectroscopy of purified NorA revealed the presence of a di-iron center, which binds oxygen in a similar way as hemerythrin. Diferrous NorA binds two molecules of NO maximally. Unexpectedly, binding of NO to the diferrous NorA required an external reductant. Two different NorA-NO species could be resolved. A minor species (up to 20%) showed an S = (1/2) EPR signal with g( perpendicular) = 2.041, and g( parallel) = 2.018, typical of a paramagnetic dinitrosyl iron complex. The major species was EPR-silent, showing characteristic signals at 420 nm and 750 nm in the optical spectrum. This species is proposed to represent a novel dinitrosyl iron complex of the form Fe(2+)-[NO](2)(2-), i.e. NO is bound as NO(-). The NO binding capacity of NorA in conjunction with its high cytoplasmic concentration (20 mum) suggests that NorA regulates transcription by lowering the free cytoplasmic concentration of NO.

Comparison of 2,4-Dichlorophenoxyacetic Acid Degradation and Plasmid Transfer in Soil Resulting from Bioaugmentation with Two Different pJP4 Donors

PubMed Central

Newby, D. T.; Gentry, T. J.; Pepper, I. L.

2000-01-01

A pilot field study was conducted to assess the impact of bioaugmentation with two plasmid pJP4-bearing microorganisms: the natural host, Ralstonia eutropha JMP134, and a laboratory-generated strain amenable to donor counterselection, Escherichia coli D11. The R. eutropha strain contained chromosomal genes necessary for mineralization of 2,4-dichlorophenoxyacetic acid (2,4-D), while the E. coli strain did not. The soil system was contaminated with 2,4-D alone or was cocontaminated with 2,4-D and Cd. Plasmid transfer to indigenous populations, plasmid persistence in soil, and degradation of 2,4-D were monitored over a 63-day period in the bioreactors. To assess the impact of contaminant reexposure, aliquots of bioreactor soil were reamended with additional 2,4-D. Both introduced donors remained culturable and transferred plasmid pJP4 to indigenous recipients, although to different extents. Isolated transconjugants were members of the Burkholderia and Ralstonia genera, suggesting multiple, if not successive, plasmid transfers. Upon a second exposure to 2,4-D, enhanced degradation was observed for all treatments, suggesting microbial adaptation to 2,4-D. Upon reexposure, degradation was most rapid for the E. coli D11-inoculated treatments. Cd did not significantly impact 2,4-D degradation or transconjugant formation. This study demonstrated that the choice of donor microorganism might be a key factor to consider for bioaugmentation efforts. In addition, the establishment of an array of stable indigenous plasmid hosts at sites with potential for reexposure or long-term contamination may be particularly useful. PMID:10919798

Interfacial microstructure and mechanical properties of Cf/AZ91D composites with TiO2 and PyC fiber coatings.

PubMed

Li, Shaolin; Qi, Lehua; Zhang, Ting; Ju, Luyan; Li, Hejun

2017-10-01

In spite of the effectiveness of the fiber coatings on interface modification of carbon fiber reinforced magnesium matrix composites, the cost and exclusive equipment for the coatings preparation are usually ignored during research work. In this paper, pyrolytic carbon (PyC) and TiO 2 were coated on carbon fiber surface to study the effects of fiber coatings on interfacial microstructure and mechanical properties of carbon fiber reinforced AZ91D composites (C f /AZ91D composites). It was indicated that both the two coatings could modify the interface and improve the mechanical properties of the composites. The ultimate tensile strength of the TiO 2 -C f /AZ91D and the PyC-C f /AZ91D composite were 333MPa and 400MPa, which were improved by 41.7% and 70.2% respectively, compared with the untreated-C f /AZ91D composite. The microstructure observation revealed that the strengthening of the composites relied on fiber integrity and moderate interfacial bonding. MgO nano-particles were generated at the interface due to the reaction of TiO 2 with Mg in the TiO 2 -C f /AZ91D composite. The volume expansion resulting from the reaction let to disordered intergranular films and crystal defects at the interface. The fibers were protected and the interfacial reaction was restrained by PyC coating in the PyC-C f /AZ91D composite. The principle to select the coating of fiber was proposed by comparing the effectiveness and cost of the coatings. Copyright © 2017 Elsevier Ltd. All rights reserved.

Two objects in Neptune's 9:1 resonance -- implications for resonance sticking in the scattering population

NASA Astrophysics Data System (ADS)

Volk, Kathryn; Murray-Clay, Ruth; Gladman, Brett; Lawler, Samantha; Yu, Tze Yeung Mathew; Alexandersen, Mike; Bannister, Michele; Chen, Ying-Yung; Dawson, Rebekah; Greenstreet, Sarah; Gwyn, Stephen; Kavelaars, J. J.; Lin, Hsing Wen; Lykawka, Patryk; Petit, Jean-Marc

2018-04-01

We discuss the detection in the Outer Solar System Origins Survey (OSSOS) of two objects in Neptune's distant 9:1 mean motion resonance at semimajor axis a≈130 au. Both objects are securely resonant on 10 Myr timescales, with one securely in the 9:1 resonance's leading asymmetric libration island and the other in either the symmetric or trailing asymmetric island. These two objects are the largest semimajor axis objects known with secure resonant classifications, and their detection in a carefully characterized survey allows for the first robust population estimate for a resonance beyond 100 au. The detection of these two objects implies a population in the 9:1 resonance of 1.1×104 objects with Hr<8.66 (D > 100 km) on similar orbits, with 95% confidence range of ∼0.4‑3×104. Integrations over 4 Gyr of an ensemble of clones chosen from within the orbit fit uncertainties for these objects reveal that they both have median resonance occupation timescales of ∼1 Gyr. These timescales are consistent with the hypothesis that these two objects originate in the scattering population but became transiently stuck to Neptune's 9:1 resonance within the last ∼1 Gyr of solar system evolution. Based on simulations of a model of the current scattering population, we estimate the expected resonance sticking population in the 9:1 resonance to be 1000--5000 objects with Hr<8.66 this is marginally consistent with the OSSOS 9:1 population estimate. We conclude that resonance sticking is a plausible explanation for the observed 9:1 population, but we also discuss the possibility of a primordial 9:1 population, which would have interesting implications for the Kuiper belt's dynamical history.

OSSOS. IX. Two Objects in Neptune's 9:1 Resonance—Implications for Resonance Sticking in the Scattering Population

NASA Astrophysics Data System (ADS)

Volk, Kathryn; Murray-Clay, Ruth A.; Gladman, Brett J.; Lawler, Samantha M.; Yu, Tze Yeung Mathew; Alexandersen, Mike; Bannister, Michele T.; Chen, Ying-Tung; Dawson, Rebekah I.; Greenstreet, Sarah; Gwyn, Stephen D. J.; Kavelaars, J. J.; Lin, Hsing Wen; Lykawka, Patryk Sofia; Petit, Jean-Marc

2018-06-01

We discuss the detection in the Outer Solar System Origins Survey (OSSOS) of two objects in Neptune’s distant 9:1 mean motion resonance at semimajor axis a ≈ 130 au. Both objects are securely resonant on 10 Myr timescales, with one securely in the 9:1 resonance’s leading asymmetric libration island and the other in either the symmetric or trailing asymmetric island. These objects are the largest semimajor axis objects with secure resonant classifications, and their detection in a carefully characterized survey allows for the first robust resonance population estimate beyond 100 au. The detection of these objects implies a 9:1 resonance population of 1.1 × 104 objects with H r < 8.66 (D ≳ 100 km) on similar orbits (95% confidence range of ∼(0.4–3) × 104). Integrations over 4 Gyr of an ensemble of clones spanning these objects’ orbit-fit uncertainties reveal that they both have median resonance occupation timescales of ∼1 Gyr. These timescales are consistent with the hypothesis that these objects originate in the scattering population but became transiently stuck to Neptune’s 9:1 resonance within the last ∼1 Gyr of solar system evolution. Based on simulations of a model of the current scattering population, we estimate the expected resonance sticking population in the 9:1 resonance to be 1000–4500 objects with H r < 8.66 this is marginally consistent with the OSSOS 9:1 population estimate. We conclude that resonance sticking is a plausible explanation for the observed 9:1 population, but we also discuss the possibility of a primordial 9:1 population, which would have interesting implications for the Kuiper Belt’s dynamical history.

Linkage of the Nit1C gene cluster to bacterial cyanide assimilation as a nitrogen source.

PubMed

Jones, Lauren B; Ghosh, Pallab; Lee, Jung-Hyun; Chou, Chia-Ni; Kunz, Daniel A

2018-05-21

A genetic linkage between a conserved gene cluster (Nit1C) and the ability of bacteria to utilize cyanide as the sole nitrogen source was demonstrated for nine different bacterial species. These included three strains whose cyanide nutritional ability has formerly been documented (Pseudomonas fluorescens Pf11764, Pseudomonas putida BCN3 and Klebsiella pneumoniae BCN33), and six not previously known to have this ability [Burkholderia (Paraburkholderia) xenovorans LB400, Paraburkholderia phymatum STM815, Paraburkholderia phytofirmans PsJN, Cupriavidus (Ralstonia) eutropha H16, Gluconoacetobacter diazotrophicus PA1 5 and Methylobacterium extorquens AM1]. For all bacteria, growth on or exposure to cyanide led to the induction of the canonical nitrilase (NitC) linked to the gene cluster, and in the case of Pf11764 in particular, transcript levels of cluster genes (nitBCDEFGH) were raised, and a nitC knock-out mutant failed to grow. Further studies demonstrated that the highly conserved nitB gene product was also significantly elevated. Collectively, these findings provide strong evidence for a genetic linkage between Nit1C and bacterial growth on cyanide, supporting use of the term cyanotrophy in describing what may represent a new nutritional paradigm in microbiology. A broader search of Nit1C genes in presently available genomes revealed its presence in 270 different bacteria, all contained within the domain Bacteria, including Gram-positive Firmicutes and Actinobacteria, and Gram-negative Proteobacteria and Cyanobacteria. Absence of the cluster in the Archaea is congruent with events that may have led to the inception of Nit1C occurring coincidentally with the first appearance of cyanogenic species on Earth, dating back 400-500 million years.

Formation of 2,4-D bound residues in soils: New insights into microbial metabolism.

PubMed

Botero, Liliana Rocío; Mougin, Chistian; Peñuela, Gustavo; Barriuso, Enrique

2017-04-15

The microbial contribution to the formation of bound residues in soils is studied by characterizing the metabolic activity of three microorganisms (Trametes versicolor, Fusarium solani and Ralstonia eutropha) on 14 C-2,4-dichlorophenoxyacetic acid (2,4-D) during incubation in synthetic liquid media and soil. A fractionation protocol was applied to quantify the 14 C-2,4-D that was incorporated into the biomass among biomolecular-like fractions. Successive fractionation of microbial biomass was implemented to break up and quantify the methanol/dichloromethane fraction (corresponding to the 14 C-lipid-like fraction), the trichloroacetic acid fraction (or hydrolysed 14 C-polysaccharide-like fraction) and the acid hydrolysable fraction (or the hydrolysed 14 C-protein-like fraction). Relevant differences in the 2,4-D degradation and biomass radioactivity distribution among the three microorganisms were found. The 14 C-protein-like fraction was the most consistent biomass fraction for reflecting the pesticide use capacity of the microorganisms under liquid and soil conditions. 2,4-D and its metabolite 4-chlorophenol were detected in methanol/dichloromethane and trichloroacetic acid fractions of the biomass of microorganisms exhibiting a low capacity to mineralize 2,4-D, thus proving that the microbial participation in the formation of bound residues while conserving the initial pesticide structure under natural soil conditions may be intimately associated with the lipid- and polysaccharide-like constituents. The fractionation protocol differentiates between 14 C that is incorporated into biomass as a biomolecular constituent and the pesticide or its metabolites that accumulate in the biomass and thus correspond to the stricto sensu definition of bound residues. Copyright © 2017 Elsevier B.V. All rights reserved.

Pease AFB, New Hampshire. Revised Uniform Summary of Surface Weather Observations.

DTIC Science & Technology

1985-07-05

rnw_ _9 28 Mar69 1. Same Same RO-362 Same2. Same q-,________ 10 Oct 70 1. Same AN/ GMO -20 Same Same2.m samaSmeE_____2. Same -gaf------ .. 11 Dec 70 1...95.4 P7.8 89.9 90.8 90.9 91.1 91.1 91.1 91.3 91.3 91.4 91.6 91.b 91.7 GF con I I . 91 .2 96.9 88.2 90.6 91.8 91.9 92.1 92.1 92.1 97.0 92.3 92.4 92.b 92.6...AT L AST CNj -.AY LESS THiAN 24 SI A L C > I L" AC A T -V I C /AC EXTREME VALUES S , " * T - .A T PRO . I’ )rL’SEP.A’.> ; : :.Ac AF _5. - STATION

DOE Office of Scientific and Technical Information (OSTI.GOV)

Villalba, Miryam I.; Canul-Tec, Juan C.; Luna-Martínez, Oscar D.

Light chain amyloidosis (AL) is a disease that affects vital organs by the fibrillar aggregation of monoclonal light chains. λ3r germ line is significantly implicated in this disease. In this paper, we contrasted the thermodynamic stability and aggregation propensity of 3mJL2 (nonamyloidogenic) and 3rJL2 (amyloidogenic) λ3 germ lines. Because of an inherent limitation (extremely low expression), Cys at position 34 of the 3r germ line was replaced by Tyr reaching a good expression yield. A second substitution (W91A) was introduced in 3r to obtain a better template to incorporate additional mutations. Although the single mutant (C34Y) was not fibrillogenic, themore » second mutation located at CDR3 (W91A) induced fibrillogenesis. We propose, for the first time, that CDR3 (position 91) affects the stability and fiber formation of human λ3r light chains. Using the double mutant (3rJL2/YA) as template, other variants were constructed to evaluate the importance of those substitutions into the stability and aggregation propensity of λ3 light chains. A change in position 7 (P7D) boosted 3rJL2/YA fibrillogenic properties. Modification of position 48 (I48M) partially reverted 3rJL2/YA fibril aggregation. Finally, changes at positions 8 (P8S) or 40 (P40S) completely reverted fibril formation. These results confirm the influential roles of N-terminal region (positions 7 and 8) and the loop 40–60 (positions 40 and 48) on AL. X-ray crystallography revealed that the three-dimensional topology of the single and double λ3r mutants was not significantly altered. Finally, this mutagenic approach helped to identify key regions implicated in λ3 AL.« less

Galaxy interactions in compact groups - II. Abundance and kinematic anomalies in HCG 91c

NASA Astrophysics Data System (ADS)

Vogt, Frédéric P. A.; Dopita, Michael A.; Borthakur, Sanchayeeta; Verdes-Montenegro, Lourdes; Heckman, Timothy M.; Yun, Min S.; Chambers, Kenneth C.

2015-07-01

Galaxies in Hickson Compact Group 91 (HCG 91) were observed with the WiFeS integral field spectrograph as part of our ongoing campaign targeting the ionized gas physics and kinematics inside star-forming members of compact groups. Here, we report the discovery of H II regions with abundance and kinematic offsets in the otherwise unremarkable star-forming spiral HCG 91c. The optical emission line analysis of this galaxy reveals that at least three H II regions harbour an oxygen abundance ˜0.15 dex lower than expected from their immediate surroundings and from the abundance gradient present in the inner regions of HCG 91c. The same star-forming regions are also associated with a small kinematic offset in the form of a lag of 5-10 km s-1 with respect to the local circular rotation of the gas. H I observations of HCG 91 from the Very Large Array and broad-band optical images from Pan-STARRS (Panoramic Survey Telescope And Rapid Response System) suggest that HCG 91c is caught early in its interaction with the other members of HCG 91. We discuss different scenarios to explain the origin of the peculiar star-forming regions detected with WiFeS, and show that evidence points towards infalling and collapsing extraplanar gas clouds at the disc-halo interface, possibly as a consequence of long-range gravitational perturbations of HCG 91c from the other group members. As such, HCG 91c provides evidence that some of the perturbations possibly associated with the early phase of galaxy evolution in compact groups impact the star-forming disc locally, and on sub-kpc scales.

A Novel p-Nitrophenol Degradation Gene Cluster from a Gram-Positive Bacterium, Rhodococcus opacus SAO101

PubMed Central

Kitagawa, Wataru; Kimura, Nobutada; Kamagata, Yoichi

2004-01-01

p-Nitrophenol (4-NP) is recognized as an environmental contaminant; it is used primarily for manufacturing medicines and pesticides. To date, several 4-NP-degrading bacteria have been isolated; however, the genetic information remains very limited. In this study, a novel 4-NP degradation gene cluster from a gram-positive bacterium, Rhodococcus opacus SAO101, was identified and characterized. The deduced amino acid sequences of npcB, npcA, and npcC showed identity with phenol 2-hydroxylase component B (reductase, PheA2) of Geobacillus thermoglucosidasius A7 (32%), with 2,4,6-trichlorophenol monooxygenase (TcpA) of Ralstonia eutropha JMP134 (44%), and with hydroxyquinol 1,2-dioxygenase (ORF2) of Arthrobacter sp. strain BA-5-17 (76%), respectively. The npcB, npcA, and npcC genes were cloned into pET-17b to construct the respective expression vectors pETnpcB, pETnpcA, and pETnpcC. Conversion of 4-NP was observed when a mixture of crude cell extracts of Escherichia coli containing pETnpcB and pETnpcA was used in the experiment. The mixture converted 4-NP to hydroxyquinol and also converted 4-nitrocatechol (4-NCA) to hydroxyquinol. Furthermore, the crude cell extract of E. coli containing pETnpcC converted hydroxyquinol to maleylacetate. These results suggested that npcB and npcA encode the two-component 4-NP/4-NCA monooxygenase and that npcC encodes hydroxyquinol 1,2-dioxygenase. The npcA and npcC mutant strains, SDA1 and SDC1, completely lost the ability to grow on 4-NP as the sole carbon source. These results clearly indicated that the cloned npc genes play an essential role in 4-NP mineralization in R. opacus SAO101. PMID:15262926

Impact of Fluoroquinolone Resistance Mutations on Gonococcal Fitness and In Vivo Selection for Compensatory Mutations

PubMed Central

Kunz, Anjali N.; Begum, Afrin A.; Wu, Hong; D'Ambrozio, Jonathan A.; Robinson, James M.; Shafer, William M.; Bash, Margaret C.; Jerse, Ann E.

2012-01-01

Background. Quinolone-resistant Neisseria gonorrhoeae (QRNG) arise from mutations in gyrA (intermediate resistance) or gyrA and parC (resistance). Here we tested the consequence of commonly isolated gyrA91/95 and parC86 mutations on gonococcal fitness. Methods. Mutant gyrA91/95 and parC86 alleles were introduced into wild-type gonococci or an isogenic mutant that is resistant to macrolides due to an mtrR−79 mutation. Wild-type and mutant bacteria were compared for growth in vitro and in competitive murine infection. Results. In vitro growth was reduced with increasing numbers of mutations. Interestingly, the gyrA91/95 mutation conferred an in vivo fitness benefit to wild-type and mtrR−79 mutant gonococci. The gyrA91/95, parC86 mutant, in contrast, showed a slight fitness defect in vivo, and the gyrA91/95, parC86, mtrR−79 mutant was markedly less fit relative to the parent strains. A ciprofloxacin-resistant (CipR) mutant was selected during infection with the gyrA91/95, parC86, mtrR−79 mutant in which the mtrR−79 mutation was repaired and the gyrA91 mutation was altered. This in vivo–selected mutant grew as well as the wild-type strain in vitro. Conclusions. gyrA91/95 mutations may contribute to the spread of QRNG. Further acquisition of a parC86 mutation abrogates this fitness advantage; however, compensatory mutations can occur that restore in vivo fitness and maintain CipR. PMID:22492860

Proline accumulation and its implication in cold tolerance of regenerable maize callus

DOE Office of Scientific and Technical Information (OSTI.GOV)

Duncan, D.R.; Widholm, J.M.

1987-03-01

Embryogenic callus of maize (Zea mays L.) inbreds B37wx, H99, H99/sup 3/H95, Mo17, and Pa91 accumulated proline to levels 2.1 to 2.5 times that of control callus when subjected to mannitol-induced water stress, cool temperatures (19/sup 0/C) and abscisic acid (ABA). A combination of 0.53 molar mannitol plus 0.1 millimolar ABA induced a proline accumulation to about 4.5 times that of control callus, equivalent to approximately 0.18 millimoles proline per gram fresh weight of callus. Proline accumulation was directly related to the level of mannitol in the medium. Levels of ABA greater than 1.0 micromolar were required in the mediummore » to induce proline accumulation comparable to that induced by mannitol. Mannitol and ABA levels that induced maximum accumulation of proline also inhibited callus growth and increased tolerance to cold. Proline (12 millimolar) added to culture media also increased the tolerance of callus to 4/sup 0/C. The increased cold tolerance induced by the combination of mannitol and ABA has permitted the storage of the maize inbreds A632, A634Ht, B37wx, C103DTrf, Fr27rhm, H99, Pa91, Va35, and W117Ht at 4/sup 0/C for 90 days which is more than double the typical survival time of callus. These studies show that proline accumulation increase the cold tolerance of regenerable maize callus.« less

Identification of Nematodirus species (Nematoda: Molineidae) from wild ruminants in Italy using ribosomal DNA markers.

PubMed

Gasser, R B; Rossi, L; Zhu, X

1999-11-01

The sequence of the second internal transcribed spacer of ribosomal DNA was determined for four species of Nematodirus (Nematodirus rupicaprae, Nematodirus oiratianus, Nematodirus davtiani alpinus and Nematodirus europaeus) from roe deer or alpine chamois. The second internal transcribed spacer of the four species varied in length from 228 to 236 bp, and the G + C contents ranged from 41 to 44%. While no intraspecific sequence variation was detected among multiple samples representing three of the taxa, sequence differences of 5.9-9.7% were detected among the four species, Nematodirus davtiani alpinus and N. rupicaprae were genetically most similar (94.1%), followed by N. oiratianus, N. europaeus and N. rupicaprae (91.1-91.5%), whereas N. oiratianus was genetically most different from N. davtiani alpinus. The interspecific sequence differences were exploited for the delineation of the four species by PCR-based restriction fragment length polymorphism (using two enzymes) and single-strand conformation polymorphism. The results have implications for diagnosis, epidemiology and for studying the systematics of the Nematodirinae.

Dual-Bioaugmentation Strategy To Enhance Remediation of Cocontaminated Soil

PubMed Central

Roane, T. M.; Josephson, K. L.; Pepper, I. L.

2001-01-01

Although metals are thought to inhibit the ability of microorganisms to degrade organic pollutants, several microbial mechanisms of resistance to metal are known to exist. This study examined the potential of cadmium-resistant microorganisms to reduce soluble cadmium levels to enhance degradation of 2,4-dichlorophenoxyacetic acid (2,4-D) under conditions of cocontamination. Four cadmium-resistant soil microorganisms were examined in this study. Resistant up to a cadmium concentration of 275 μg ml−1, these isolates represented the common soil genera Arthrobacter, Bacillus, and Pseudomonas. Isolates Pseudomonas sp. strain H1 and Bacillus sp. strain H9 had a plasmid-dependent intracellular mechanism of cadmium detoxification, reducing soluble cadmium levels by 36%. Isolates Arthrobacter strain D9 and Pseudomonas strain I1a both produced an extracellular polymer layer that bound and reduced soluble cadmium levels by 22 and 11%, respectively. Although none of the cadmium-resistant isolates could degrade 2,4-D, results of dual-bioaugmentation studies conducted with both pure culture and laboratory soil microcosms showed that each of four cadmium-resistant isolates supported the degradation of 500-μg ml−1 2,4-D by the cadmium-sensitive 2,4-D degrader Ralstonia eutropha JMP134. Degradation occurred in the presence of up to 24 μg of cadmium ml−1 in pure culture and up to 60 μg of cadmium g−1 in amended soil microcosms. In a pilot field study conducted with 5-gallon soil bioreactors, the dual-bioaugmentation strategy was again evaluated. Here, the cadmium-resistant isolate Pseudomonas strain H1 enhanced degradation of 2,4-D in reactors inoculated with R. eutropha JMP134 in the presence of 60 μg of cadmium g−1. Overall, dual bioaugmentation appears to be a viable approach in the remediation of cocontaminated soils. PMID:11425743

Cometabolic degradation of ethyl mercaptan by phenol-utilizing Ralstonia eutropha in suspended growth and gas-recycling trickle-bed reactor.

PubMed

Sedighi, Mahsa; Zamir, Seyed Morteza; Vahabzadeh, Farzaneh

2016-01-01

The degradability of ethyl mercaptan (EM), by phenol-utilizing cells of Ralstonia eutropha, in both suspended and immobilized culture systems, was investigated in the present study. Free-cells experiments conducted at EM concentrations ranging from 1.25 to 14.42 mg/l, showed almost complete removal of EM at concentrations below 10.08 mg/l, which is much higher than the maximum biodegradable EM concentration obtained in experiments that did not utilize phenol as the primary substrate, i.e. 2.5 mg/l. The first-order kinetic rate constant (kSKS) for EM biodegradation by the phenol-utilizing cells (1.7 l/g biomass/h) was about 10 times higher than by cells without phenol utilization. Immobilized-cells experiments performed in a gas recycling trickle-bed reactor packed with kissiris particles at EM concentrations ranging from 1.6 to 36.9 mg/l, showed complete removal at all tested concentrations in a much shorter time, compared with free cells. The first-order kinetic rate constant (rmaxKs) for EM utilization was 0.04 l/h for the immobilized system compared to 0.06 for the suspended-growth culture, due to external mass transfer diffusion. Diffusion limitation was decreased by increasing the recycling-liquid flow rate from 25 to 65 ml/min. The removed EM was almost completely mineralized according to TOC and sulfate measurements. Shut down and starvation experiments revealed that the reactor could effectively handle the starving conditions and was reliable for full-scale application. Copyright © 2015 Elsevier Ltd. All rights reserved.

Optimization of cyanophycin production in recombinant strains of Pseudomonas putida and Ralstonia eutropha employing elementary mode analysis and statistical experimental design.

PubMed

Diniz, Simone Cardoso; Voss, Ingo; Steinbüchel, Alexander

2006-03-05

Elementary mode analysis was applied to simulate conditions for cyanophycin (CGP) biosynthesis and to optimize its production in bacteria. The conclusions from these simulations were confirmed by experiments with recombinant strains of the wild types and polyhydroxyalkanoate (PHA)-negative mutants of Ralstonia eutropha and Pseudomonas putida expressing CGP synthetase genes (cphA) of Synechocystis sp. strain PCC6308 or Anabaena sp. strain PCC7120. In particular, the effects of suitable precursor substrates and of oxygen supply as well as of the capability to accumulate PHA in addition to CGP biosynthesis were investigated. Since CGP consists of the amino acids aspartate and arginine, the tricarboxylic acid cycle (TCC), which provides intermediates for biosynthesis of these amino acids, seems to be important. Excretion of intermediates of the TCC upon cultivation at restricted oxygen supply and conversion of fumarate mainly to malate and to only little succinate in the absence of oxygen indicated that TCC intermediates for arginine and aspartate biosynthesis were provided by the oxidative or reductive parts of the TCC, respectively. The following important conclusions were made from the experiments and the simulations: (i) external arginine additionally supplied to the medium, (ii) oxygen limitation, and (iii) absence of PHA accumulation exerted positive effects on CGP accumulation. These conclusions were utilized to obtain CGP contents in the cells of as high as 17.9% (w x w(-1)) during cultivation of the investigated bacteria at the 30-L scale using mineral salts medium. Such high CGP contents were previously not obtained with these bacteria at a 30-L scale, even if complex media were used.

Requirements for construction of a functional hybrid complex of photosystem I and [NiFe]-hydrogenase.

PubMed

Schwarze, Alexander; Kopczak, Marta J; Rögner, Matthias; Lenz, Oliver

2010-04-01

The development of cellular systems in which the enzyme hydrogenase is efficiently coupled to the oxygenic photosynthesis apparatus represents an attractive avenue to produce H(2) sustainably from light and water. Here we describe the molecular design of the individual components required for the direct coupling of the O(2)-tolerant membrane-bound hydrogenase (MBH) from Ralstonia eutropha H16 to the acceptor site of photosystem I (PS I) from Synechocystis sp. PCC 6803. By genetic engineering, the peripheral subunit PsaE of PS I was fused to the MBH, and the resulting hybrid protein was purified from R. eutropha to apparent homogeneity via two independent affinity chromatographical steps. The catalytically active MBH-PsaE (MBH(PsaE)) hybrid protein could be isolated only from the cytoplasmic fraction. This was surprising, since the MBH is a substrate of the twin-arginine translocation system and was expected to reside in the periplasm. We conclude that the attachment of the additional PsaE domain to the small, electron-transferring subunit of the MBH completely abolished the export competence of the protein. Activity measurements revealed that the H(2) production capacity of the purified MBH(PsaE) fusion protein was very similar to that of wild-type MBH. In order to analyze the specific interaction of MBH(PsaE) with PS I, His-tagged PS I lacking the PsaE subunit was purified via Ni-nitrilotriacetic acid affinity and subsequent hydrophobic interaction chromatography. Formation of PS I-hydrogenase supercomplexes was demonstrated by blue native gel electrophoresis. The results indicate a vital prerequisite for the quantitative analysis of the MBH(PsaE)-PS I complex formation and its light-driven H(2) production capacity by means of spectroelectrochemistry.

Genetic Variation for Thermotolerance in Lettuce Seed Germination Is Associated with Temperature-Sensitive Regulation of ETHYLENE RESPONSE FACTOR1 (ERF1)1[OPEN

PubMed Central

O’Brien, Laurel K.; Truco, Maria Jose; Huo, Heqiang; Sideman, Rebecca; Hayes, Ryan; Michelmore, Richard W.

2016-01-01

Seeds of most lettuce (Lactuca sativa) cultivars are susceptible to thermoinhibition, or failure to germinate at temperatures above approximately 28°C, creating problems for crop establishment in the field. Identifying genes controlling thermoinhibition would enable the development of cultivars lacking this trait and, therefore, being less sensitive to high temperatures during planting. Seeds of a primitive accession (PI251246) of lettuce exhibited high-temperature germination capacity up to 33°C. Screening a recombinant inbred line population developed from PI215246 and cv Salinas identified a major quantitative trait locus (Htg9.1) from PI251246 associated with the high-temperature germination phenotype. Further genetic analyses discovered a tight linkage of the Htg9.1 phenotype with a specific DNA marker (NM4182) located on a single genomic sequence scaffold. Expression analyses of the 44 genes encoded in this genomic region revealed that only a homolog of Arabidopsis (Arabidopsis thaliana) ETHYLENE RESPONSE FACTOR1 (termed LsERF1) was differentially expressed between PI251246 and cv Salinas seeds imbibed at high temperature (30°C). LsERF1 belongs to a large family of transcription factors associated with the ethylene-signaling pathway. Physiological assays of ethylene synthesis, response, and action in parental and near-isogenic Htg9.1 genotypes strongly implicate LsERF1 as the gene responsible for the Htg9.1 phenotype, consistent with the established role for ethylene in germination thermotolerance of Compositae seeds. Expression analyses of genes associated with the abscisic acid and gibberellin biosynthetic pathways and results of biosynthetic inhibitor and hormone response experiments also support the hypothesis that differential regulation of LsERF1 expression in PI251246 seeds elevates their upper temperature limit for germination through interactions among pathways regulated by these hormones. Our results support a model in which LsERF1 acts through the promotion of gibberellin biosynthesis to counter the inhibitory effects of abscisic acid and, therefore, promote germination at high temperatures. PMID:26574598

Selective Sweep Analysis in the Genomes of the 91-R and 91-C Drosophila melanogaster Strains Reveals Few of the ‘Usual Suspects’ in Dichlorodiphenyltrichloroethane (DDT) Resistance

PubMed Central

Steele, Laura D.; Coates, Brad; Valero, M. Carmen; Sun, Weilin; Seong, Keon Mook; Muir, William M.; Clark, John M.; Pittendrigh, Barry R.

2015-01-01

Adaptation of insect phenotypes for survival after exposure to xenobiotics can result from selection at multiple loci with additive genetic effects. To the authors’ knowledge, no selective sweep analysis has been performed to identify such loci in highly dichlorodiphenyltrichloroethane (DDT) resistant insects. Here we compared a highly DDT resistant phenotype in the Drosophila melanogaster (Drosophila) 91-R strain to the DDT susceptible 91-C strain, both of common origin. Whole genome re-sequencing data from pools of individuals was generated separately for 91-R and 91-C, and mapped to the reference Drosophila genome assembly (v. 5.72). Thirteen major and three minor effect chromosome intervals with reduced nucleotide diversity (π) were identified only in the 91-R population. Estimates of Tajima's D (D) showed corresponding evidence of directional selection in these same genome regions of 91-R, however, no similar reductions in π or D estimates were detected in 91-C. An overabundance of non-synonymous proteins coding to synonymous changes were identified in putative open reading frames associated with 91-R. Except for NinaC and Cyp4g1, none of the identified genes were the ‘usual suspects’ previously observed to be associated with DDT resistance. Additionally, up-regulated ATP-binding cassette transporters have been previously associated with DDT resistance; however, here we identified a structurally altered MDR49 candidate resistance gene. The remaining fourteen genes have not previously been shown to be associated with DDT resistance. These results suggest hitherto unknown mechanisms of DDT resistance, most of which have been overlooked in previous transcriptional studies, with some genes having orthologs in mammals. PMID:25826265

26 CFR 31.3306(c)(9)-1 - Railroad industry; services performed by an employee or an employee representative under the...

Code of Federal Regulations, 2010 CFR

2010-04-01

... employee or an employee representative under the Railroad Unemployment Insurance Act. 31.3306(c)(9)-1... SOURCE Federal Unemployment Tax Act (Chapter 23, Internal Revenue Code of 1954) § 31.3306(c)(9)-1... Unemployment Insurance Act. (a) Services performed by an individual as an “employee” or as an “employee...

26 CFR 31.3306(c)(9)-1 - Railroad industry; services performed by an employee or an employee representative under the...

Code of Federal Regulations, 2011 CFR

2011-04-01

... employee or an employee representative under the Railroad Unemployment Insurance Act. 31.3306(c)(9)-1... SOURCE Federal Unemployment Tax Act (Chapter 23, Internal Revenue Code of 1954) § 31.3306(c)(9)-1... Unemployment Insurance Act. (a) Services performed by an individual as an “employee” or as an “employee...

26 CFR 31.3306(c)(9)-1 - Railroad industry; services performed by an employee or an employee representative under the...

Code of Federal Regulations, 2014 CFR

2014-04-01

... employee or an employee representative under the Railroad Unemployment Insurance Act. 31.3306(c)(9)-1... SOURCE Federal Unemployment Tax Act (Chapter 23, Internal Revenue Code of 1954) § 31.3306(c)(9)-1... Unemployment Insurance Act. (a) Services performed by an individual as an “employee” or as an “employee...

26 CFR 31.3306(c)(9)-1 - Railroad industry; services performed by an employee or an employee representative under the...

Code of Federal Regulations, 2012 CFR

2012-04-01

... employee or an employee representative under the Railroad Unemployment Insurance Act. 31.3306(c)(9)-1... SOURCE Federal Unemployment Tax Act (Chapter 23, Internal Revenue Code of 1954) § 31.3306(c)(9)-1... Unemployment Insurance Act. (a) Services performed by an individual as an “employee” or as an “employee...

26 CFR 31.3306(c)(9)-1 - Railroad industry; services performed by an employee or an employee representative under the...

Code of Federal Regulations, 2013 CFR

2013-04-01

... employee or an employee representative under the Railroad Unemployment Insurance Act. 31.3306(c)(9)-1... SOURCE Federal Unemployment Tax Act (Chapter 23, Internal Revenue Code of 1954) § 31.3306(c)(9)-1... Unemployment Insurance Act. (a) Services performed by an individual as an “employee” or as an “employee...

Polyhydroxyalkanoate (PHA) Granules Have no Phospholipids

PubMed Central

Bresan, Stephanie; Sznajder, Anna; Hauf, Waldemar; Forchhammer, Karl; Pfeiffer, Daniel; Jendrossek, Dieter

2016-01-01

Polyhydroxybutyrate (PHB) granules, also designated as carbonosomes, are supra-molecular complexes in prokaryotes consisting of a PHB polymer core and a surface layer of structural and functional proteins. The presence of suspected phospholipids in the surface layer is based on in vitro data of isolated PHB granules and is often shown in cartoons of the PHB granule structure in reviews on PHB metabolism. However, the in vivo presence of a phospholipid layer has never been demonstrated. We addressed this topic by the expression of fusion proteins of DsRed2EC and other fluorescent proteins with the phospholipid-binding domain (LactC2) of lactadherin in three model organisms. The fusion proteins specifically localized at the cell membrane of Ralstonia eutropha but did not co-localize with PHB granules. The same result was obtained for Pseudomonas putida, a species that accumulates another type of polyhydroxyalkanoate (PHA) granules related to PHB. Notably, DsRed2EC-LactC2 expressed in Magnetospirillum gryphiswaldense was detected at the position of membrane-enclosed magnetosome chains and at the cytoplasmic membrane but not at PHB granules. In conclusion, the carbonosomes of representatives of α-proteobacteria, β-proteobacteria and γ-proteobacteria have no phospholipids in vivo and we postulate that the PHB/PHA granule surface layers in natural producers generally are free of phospholipids and consist of proteins only. PMID:27222167

Cross-border drug injection relationships among injection drug users in Tijuana, Mexico

PubMed Central

Wagner, Karla D.; Pollini, Robin A.; Patterson, Thomas L.; Lozada, Remedios; Ojeda, Victoria D.; Brouwer, Kimberly C.; Vera, Alicia; Volkmann, Tyson A.; Strathdee, Steffanie A.

2010-01-01

Background International borders are unique social and environmental contexts characterized by high levels of mobility. Among drug users, mobility increases risk for human immunodeficiency virus (HIV) in part through its effects on the social environment. However, the social dynamics of drug users living in border regions are understudied. Methods 1056 injection drug users (IDUs) residing in Tijuana, Mexico were recruited using respondent-driven sampling (RDS) from 2006 to 2007, and underwent surveys and testing for HIV, syphilis, and tuberculosis (TB). Using logistic regression on baseline data, we identified correlates of having ever injected drugs with someone from the US. Results Almost half (48%) reported ever injecting drugs with someone from the US. In RDS-adjusted logistic regression, factors independently associated with having ever injected with someone from the US included: having greater than middle school education (Adjusted Odds Ratio [AOR] 2.91; 95% Confidence Interval [C.I.] 1.52, 5.91), speaking English (AOR 3.24, 95% C.I. 1.96, 5.36), age (AOR 1.10 per year; 95% C.I. 1.07, 1.14), age at initiation of injection drug use (AOR 0.90 per year; 95% C.I. 0.86, 0.94), homelessness (AOR 2.61; 95% C.I. 1.27, 5.39), and having ever been incarcerated (AOR 11.82; 95% C.I., 5.22, 26.77). No associations with HIV, syphilis, TB, drug use, or injection risk behavior were detected. Conclusion Findings suggest that IDU networks in Mexico and the US may transcend international borders, with implications for cross-border transmission of infectious disease. Binational programs and policies need to consider the structure and geographic distribution of drug using networks. PMID:20889270

U-Pb SHRIMP geochronology of zircon in garnet peridotite from the Sulu UHP terrane, China: Implications for mantle metasomatism and subduction-zone UHP metamorphism

USGS Publications Warehouse

Zhang, R.Y.; Yang, J.S.; Wooden, J.L.; Liou, J.G.; Li, T.F.

2005-01-01

We studied the Zhimafang ultrahigh-pressure metamorphic (UHP) peridotite from pre-pilot drill hole PP-1 of Chinese Continental Scientific Drilling project in the Sulu UHP terrane, eastern China. The peridotite occurs as lens within quartofeldspathic gneiss, and has an assemblage of Ol + Opx + Cpx + Phl + Ti-clinohumite (Ti-Chu) + Grt (or chromite) ?? magnesite (Mgs). Zircons were separated from cores at depths of 152 m (C24, garnet lhezolite), 160 m (C27, strongly retrograded phlogopite-rich peridotite) and 225 m (C50, banded peridotite), and were dated by SHRIMP mass spectrometer. Isometric zircons without inherited cores contain inclusions of olivine (Fo91-92), enstatite (En91-92), Ti-clinohumite, diopside, phlogopite and apatite. The enstatite inclusions have low Al2O3 contents of only 0.04-0.13 wt.%, indicating a UHP metamorphic origin. The weighted mean 206Pb/238U zircon age for garnet lherzolite (C24) is 221 ?? 3 Ma, and a discordia lower intercept age for peridotite (C50) is 220 ?? 2 Ma. These ages are within error and represent the time of subduction-zone UHP metamorphism. A younger lower intercept age of 212 ?? 3 Ma for a foliated wehrlite (C27) was probably caused by Pb loss during retrograde metamorphism. The source of zirconium may be partially attributed to melt/fluid metasomatism within the mantle wedge. Geochronological and geochemical data confirm that the mantle-derived Zhimafang garnet peridotites (probably the most representative type of Sulu garnet peridotites) were tectonically inserted into a subducting crustal slab and subjected to in situ Triassic subduction-zone UHP metamorphism. ?? 2005 Elsevier B.V. All rights reserved.

14 CFR Appendix C to Part 91 - Operations in the North Atlantic (NAT) Minimum Navigation Performance Specifications (MNPS) Airspace

Code of Federal Regulations, 2014 CFR

2014-01-01

... OPERATING RULES GENERAL OPERATING AND FLIGHT RULES Pt. 91, App. C Appendix C to Part 91—Operations in the... Oceanic Control Area, excluding the areas west of 60 degrees west and south of 38 degrees 30 minutes north... shall be less than 6.3 NM (11.7 Km). Standard deviation is a statistical measure of data about a mean...

14 CFR Appendix C to Part 91 - Operations in the North Atlantic (NAT) Minimum Navigation Performance Specifications (MNPS) Airspace

Code of Federal Regulations, 2013 CFR

2013-01-01

... OPERATING RULES GENERAL OPERATING AND FLIGHT RULES Pt. 91, App. C Appendix C to Part 91—Operations in the... Oceanic Control Area, excluding the areas west of 60 degrees west and south of 38 degrees 30 minutes north... shall be less than 6.3 NM (11.7 Km). Standard deviation is a statistical measure of data about a mean...

14 CFR Appendix C to Part 91 - Operations in the North Atlantic (NAT) Minimum Navigation Performance Specifications (MNPS) Airspace

Code of Federal Regulations, 2011 CFR

2011-01-01

... OPERATING RULES GENERAL OPERATING AND FLIGHT RULES Pt. 91, App. C Appendix C to Part 91—Operations in the... Oceanic Control Area, excluding the areas west of 60 degrees west and south of 38 degrees 30 minutes north... shall be less than 6.3 NM (11.7 Km). Standard deviation is a statistical measure of data about a mean...

14 CFR Appendix C to Part 91 - Operations in the North Atlantic (NAT) Minimum Navigation Performance Specifications (MNPS) Airspace

Code of Federal Regulations, 2012 CFR

2012-01-01

... OPERATING RULES GENERAL OPERATING AND FLIGHT RULES Pt. 91, App. C Appendix C to Part 91—Operations in the... Oceanic Control Area, excluding the areas west of 60 degrees west and south of 38 degrees 30 minutes north... shall be less than 6.3 NM (11.7 Km). Standard deviation is a statistical measure of data about a mean...

Influence of native plasmids to fitness of Pantoea vagans strain C9-1

USDA-ARS?s Scientific Manuscript database

Pantoea vagans strain C9-1 is a biological control agent for fire blight caused by Erwinia amylovora. We cured C9-1 of two of its three plasmids: pPag2, pPag3, and both pPag2 and pPag3, tested phenotypes of the derivatives, and evaluated blossom colonization in the field. pPag2 (166 kb) encodes for ...

Influence of endogenous plasmids on phenotypes of Pantoea vagans strain C9-1 associated with epiphytic fitness

USDA-ARS?s Scientific Manuscript database

Pantoea vagans strain C9-1 is an effective biological control agent for fire blight of pear and apple. C9-1 carries three circular plasmids: pPag1 (168 kb), pPag2 (166 kb), and pPag3 (530 kb). Of these, pPag3, a member of the large Pantoea plasmid family, was proposed to contribute to epiphytic fitn...

Provenance of Miocene submarine fans in the Shikoku Basin: Results from NanTroSEIZE and implications for stratigraphic correlation of subduction inputs

NASA Astrophysics Data System (ADS)

Pickering, K. T.; Underwood, M.; Moore, G. F.

2013-12-01

Seismo-stratigraphy, coring and LWD during IODP Expeditions 319, 322, and 333 (Sites C0011 / C0012) show three Miocene submarine fans in the NE Shikoku Basin, with broadly coeval deposits at ODP Site 1177 and DSDP Site 297, NW Shikoku Basin. Pickering et al. (2013) have shown that the sediment dispersal patterns for these fans have major implications for paleogeographies at that time. The oldest, Middle Miocene Kyushu Fan is the finest grained, has a sheet-like geometry, and was fed by quartz-rich sediment gravity-flows derived mostly from an ancestral landmass in the East China Sea. This likely sediment provenance is further supported by U-Pb zircon and fission track analysis of both zircons and apatites from sediments taken from the forearc and trench of the Nankai Trough, together with rivers from southwest Japan, that point to the influence of the Yangtze River in supplying into the Shikoku Basin prior to rifting of the Okinawa Trough at 10 to 6 Ma (Clift et al. 2013). During prolonged hemipelagic mud deposition at C0011-C0012 (12.2 to 9.1 Ma), sand supply continued at Sites 1177 and 297. Sand delivery to much of the Shikoku Basin, however, probably halted during a phase of sinistral strike-slip and oblique plate motion, after which the Daiichi Zenisu Fan (9.1 to 8.0 Ma) was fed by submarine channels. The youngest fan (Daini Zenisu; 8.0 to 7.6 Ma) has sheet-like geometry with thick-bedded, coarse-grained pumiceous sandstones. The pumice fragments were fed from a mixed provenance that included the collision zone of the Izu-Bonin and Honshu arcs. The shift from channelized to sheet-like flows was probably favored by renewal of relatively rapid northward subduction, which accentuated the trench as a bathymetric depression. Understanding the stratigraphic position and 3-D geometry of the sandbodies has important implications for stratigraphic correlation throughout the northern Shikoku Basin, together with subduction-related processes, including the potential for focused fluid flow and fluid overpressures above and below the plate-boundary fault. References Pickering, K.T., Underwood, M.B., Saito, S., Naruse, H., Kutterolf, S., Scudder, R., Park, J.-O., Moore, G.F. & Slagle, A. 2013. Geochemistry, Geophysics, Geosystems, 14, doi:10.1002/ggge.20107 Clift, P.D., Carter, A., Nicholson, U. & Masago, H. 2013. Tectonics, doi: 10.1002/tect.20033

From Waste to Plastic: Synthesis of Poly(3-Hydroxypropionate) in Shimwellia blattae

PubMed Central

Heinrich, Daniel; Andreessen, Björn; Madkour, Mohamed H.; Al-Ghamdi, Mansour A.; Shabbaj, Ibrahim I.

2013-01-01

In recent years, glycerol has become an attractive carbon source for microbial processes, as it accumulates massively as a by-product of biodiesel production, also resulting in a decline of its price. A potential use of glycerol in biotechnology is the synthesis of poly(3-hydroxypropionate) [poly(3HP)], a biopolymer with promising properties which is not synthesized by any known wild-type organism. In this study, the genes for 1,3-propanediol dehydrogenase (dhaT) and aldehyde dehydrogenase (aldD) of Pseudomonas putida KT2442, propionate-coenzyme A (propionate-CoA) transferase (pct) of Clostridium propionicum X2, and polyhydroxyalkanoate (PHA) synthase (phaC1) of Ralstonia eutropha H16 were cloned and expressed in the 1,3-propanediol producer Shimwellia blattae. In a two-step cultivation process, recombinant S. blattae cells accumulated up to 9.8% ± 0.4% (wt/wt [cell dry weight]) poly(3HP) with glycerol as the sole carbon source. Furthermore, the engineered strain tolerated the application of crude glycerol derived from biodiesel production, yielding a cell density of 4.05 g cell dry weight/liter in a 2-liter fed-batch fermentation process. PMID:23542629

From waste to plastic: synthesis of poly(3-hydroxypropionate) in Shimwellia blattae.

PubMed

Heinrich, Daniel; Andreessen, Björn; Madkour, Mohamed H; Al-Ghamdi, Mansour A; Shabbaj, Ibrahim I; Steinbüchel, Alexander

2013-06-01

In recent years, glycerol has become an attractive carbon source for microbial processes, as it accumulates massively as a by-product of biodiesel production, also resulting in a decline of its price. A potential use of glycerol in biotechnology is the synthesis of poly(3-hydroxypropionate) [poly(3HP)], a biopolymer with promising properties which is not synthesized by any known wild-type organism. In this study, the genes for 1,3-propanediol dehydrogenase (dhaT) and aldehyde dehydrogenase (aldD) of Pseudomonas putida KT2442, propionate-coenzyme A (propionate-CoA) transferase (pct) of Clostridium propionicum X2, and polyhydroxyalkanoate (PHA) synthase (phaC1) of Ralstonia eutropha H16 were cloned and expressed in the 1,3-propanediol producer Shimwellia blattae. In a two-step cultivation process, recombinant S. blattae cells accumulated up to 9.8% ± 0.4% (wt/wt [cell dry weight]) poly(3HP) with glycerol as the sole carbon source. Furthermore, the engineered strain tolerated the application of crude glycerol derived from biodiesel production, yielding a cell density of 4.05 g cell dry weight/liter in a 2-liter fed-batch fermentation process.

10 CFR 9.1 - Scope and purpose.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 10 Energy 1 2014-01-01 2014-01-01 false Scope and purpose. 9.1 Section 9.1 Energy NUCLEAR... Freedom of Information Act, 5 U.S.C. 552, concerning the availability to the public of Nuclear Regulatory... 1974, 5 U.S.C. 552a, concerning disclosure and availability of certain Nuclear Regulatory Commission...

10 CFR 9.1 - Scope and purpose.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 10 Energy 1 2012-01-01 2012-01-01 false Scope and purpose. 9.1 Section 9.1 Energy NUCLEAR... Freedom of Information Act, 5 U.S.C. 552, concerning the availability to the public of Nuclear Regulatory... 1974, 5 U.S.C. 552a, concerning disclosure and availability of certain Nuclear Regulatory Commission...

10 CFR 9.1 - Scope and purpose.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 10 Energy 1 2013-01-01 2013-01-01 false Scope and purpose. 9.1 Section 9.1 Energy NUCLEAR... Freedom of Information Act, 5 U.S.C. 552, concerning the availability to the public of Nuclear Regulatory... 1974, 5 U.S.C. 552a, concerning disclosure and availability of certain Nuclear Regulatory Commission...

10 CFR 9.1 - Scope and purpose.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 10 Energy 1 2010-01-01 2010-01-01 false Scope and purpose. 9.1 Section 9.1 Energy NUCLEAR... Freedom of Information Act, 5 U.S.C. 552, concerning the availability to the public of Nuclear Regulatory... 1974, 5 U.S.C. 552a, concerning disclosure and availability of certain Nuclear Regulatory Commission...

10 CFR 9.1 - Scope and purpose.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 10 Energy 1 2011-01-01 2011-01-01 false Scope and purpose. 9.1 Section 9.1 Energy NUCLEAR... Freedom of Information Act, 5 U.S.C. 552, concerning the availability to the public of Nuclear Regulatory... 1974, 5 U.S.C. 552a, concerning disclosure and availability of certain Nuclear Regulatory Commission...

Cyclic AMP- and (Rp)-cAMPS-induced Conformational Changes in a Complex of the Catalytic and Regulatory (RIα) Subunits of Cyclic AMP-dependent Protein Kinase*

PubMed Central

Anand, Ganesh S.; Krishnamurthy, Srinath; Bishnoi, Tanushree; Kornev, Alexandr; Taylor, Susan S.; Johnson, David A.

2010-01-01

We took a discovery approach to explore the actions of cAMP and two of its analogs, one a cAMP mimic ((Sp)-adenosine cyclic 3′:5′-monophosphorothioate ((Sp)-cAMPS)) and the other a diastereoisomeric antagonist ((Rp)-cAMPS), on a model system of the type Iα cyclic AMP-dependent protein kinase holoenzyme, RIα(91–244)·C-subunit, by using fluorescence spectroscopy and amide H/2H exchange mass spectrometry. Specifically, for the fluorescence experiments, fluorescein maleimide was conjugated to three cysteine single residue substitution mutants, R92C, T104C, and R239C, of RIα(91–244), and the effects of cAMP, (Sp)-cAMPS, and (Rp)-cAMPS on the kinetics of R-C binding and the time-resolved anisotropy of the reporter group at each conjugation site were measured. For the amide exchange experiments, ESI-TOF mass spectrometry with pepsin proteolytic fragmentation was used to assess the effects of (Rp)-cAMPS on amide exchange of the RIα(91–244)·C-subunit complex. We found that cAMP and its mimic perturbed at least parts of the C-subunit interaction Sites 2 and 3 but probably not Site 1 via reduced interactions of the linker region and αC of RIα(91–244). Surprisingly, (Rp)-cAMPS not only increased the affinity of RIα(91–244) toward the C-subunit by 5-fold but also produced long range effects that propagated through both the C- and R-subunits to produce limited unfolding and/or enhanced conformational flexibility. This combination of effects is consistent with (Rp)-cAMPS acting by enhancing the internal entropy of the R·C complex. Finally, the (Rp)-cAMPS-induced increase in affinity of RIα(91–244) toward the C-subunit indicates that (Rp)-cAMPS is better described as an inverse agonist because it decreases the fractional dissociation of the cyclic AMP-dependent protein kinase holoenzyme and in turn its basal activity. PMID:20167947

Electrochemical properties of composite cathodes using Sm doped layered perovskite for intermediate temperature-operating solid oxide fuel cell

NASA Astrophysics Data System (ADS)

Baek, Seung-Wook; Azad, Abul K.; Irvine, John T. S.; Choi, Won Seok; Kang, Hyunil; Kim, Jung Hyun

2018-02-01

SmBaCo2O5+d (SBCO) showed the lowest observed Area Specific Resistance (ASR) value in the LnBaCo2O5+d (Ln: Pr, Nd, Sm, and Gd) oxide system for the overall temperature ranges tested. The ASR of a composite cathode (mixture of SBCO and Ce0.9Gd0.1O2-d) on a Ce0.9Gd0.1O2-d (CGO91) electrolyte decreased with respect to the CGO91 content; the percolation limit was also achieved for a 50 wt% SBCO and 50 wt% CGO91 (SBCO50) composite cathode. The ASRs of SBCO50 on the dense CGO91 electrolyte in the overall temperature range of 500-750 °C were relatively lower than those of SBCO50 on the CGO91 coated dense 8 mol% yttria-stabilized zirconia (8YSZ) electrolyte for the same temperature range. From 750 °C and for all higher temperatures tested, however, the ASRs of SBCO50 on the CGO91 coated dense 8YSZ electrolyte were lower than those of the CGO91 electrolyte. The maximum power densities of SBCO50 on the Ni-8YSZ/8YSZ/CGO91 buffer layer were 1.034 W cm-2 and 0.611 W cm-2 at 800 °C and 700 °C.

In vitro metabolism of phenytoin in 36 CYP2C9 variants found in the Chinese population.

PubMed

Chen, Lian-Guo; Wang, Zhe; Zhu, Yuan; Xiong, Jian-Hua; Sun, Li-Rong; Dai, Da-Peng; Cai, Jian-Ping; Hu, Guo-Xin

2016-06-25

Cytochrome P450 2C9 (CYP2C9) is an important member of the cytochrome P450 enzyme superfamily, with 57 CYP2C9 allelic variants being previously reported. Recently, we identified 22 novel alleles (*36 -*56 and N418T) in the Han Chinese population. This study aims to assess the catalytic activities of wild-type (CYP2C9*1) and 36 CYP2C9 allelic variants found in the Chinese population toward phenytoin (PHT) in vitro. Insect microsomes expressing CYP2C9*1 and 36 CYP2C9 variants were incubated with 1-200 μM phenytoin for 30 min at 37 °C. Then, these products were extracted and the signal detection was performed by HPLC-MS/MS. The intrinsic clearance (Vmax/Km) values of all variants, with the exception of CYP2C9*2, CYP2C9*11, CYP2C9*23, CYP2C9*29, CYP2C9*34, CYP2C9*38, CYP2C9*44, CYP2C9*46 and CYP2C9*48, were significantly different from CYP2C9*1. CYP2C9*27, *40, *41, *47, *49, *51, *53, *54, *56 and N418T variant exhibited markedly larger values than CYP2C9*1 (>152.8%), whereas 17 variants exhibited smaller values (from 48.6% to 99.9%) due to larger Km and/or smaller Vmax values than CYP2C9*1. The findings suggest that more attention should be paid on subjects carrying these infrequent CYP2C9 alleles when administering phenytoin in clinic. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Desert Express: An Analysis on Improved Customer Service

DTIC Science & Technology

1991-09-01

Nt MARQ 3,199 Of. DESERT EXPRESS: AN ANALYSIS ON IMPROVED CUSTOMER SERVICE THESIS Thomas C Thaiheim, Majo-,r USAF AFTT/GLM/LSM,/91S-64 ?Z; W...Astq vt.: tyc a l AFIT/GLM/,LSM/91S-64 DESERT EXPRESS: AN ANALYSIS ON IMPROVED CUSTOMER SERVICE THESIS Thomas C. Thalheim, Major, USAF AFIT/GLM/LSM...91S-64 Approved for public release; distribution unlimited AFIT/GLM/LSM/91S-64 DESERT EXPRESS: AN ANALYSIS ON IMPROVED CUSTOMER SERVICE THESIS

Polyhydroxyalkanoate production by a novel bacterium Massilia sp. UMI-21 isolated from seaweed, and molecular cloning of its polyhydroxyalkanoate synthase gene.

PubMed

Han, Xuerong; Satoh, Yasuharu; Kuriki, Yumi; Seino, Teruyuki; Fujita, Shinji; Suda, Takanori; Kobayashi, Takanori; Tajima, Kenji

2014-11-01

We successfully isolated one microorganism (UMI-21) from Ulva, a green algae that contains starch. The strain UMI-21 can produce polyhydroxyalkanoate (PHA) from starch, maltotriose, or maltose as a sole carbon source. Taxonomic studies and 16S rDNA sequence analysis revealed that strain UMI-21 was phylogenetically related to species of the genus Massilia. The PHA content under the cultivation condition using a 10-L jar fermentor was 45.5% (w/w). This value was higher than that obtained after cultivation in a flask, suggesting the possibility of large-scale PHA production by UMI-21 from starch. A major issue for the industrial production of microbial PHAs is the very high production cost. Starch is a relatively inexpensive substrate that is also found in abundant seaweeds such as Ulva. Therefore, the strain isolated in this study may be very useful for producing PHA from seaweeds containing polysaccharides such as starch. In addition, a 3.7-kbp DNA fragment containing the whole PHA synthase gene (phaC) was obtained from the strain UMI-21. The results of open reading frame (ORF) analysis suggested that the DNA fragment contained two ORFs, which were composed of 1740 (phaC) and 564 bp (phaR). The deduced amino acid sequence of PhaC from strain UMI-21 shared high similarity with PhaC from Ralstonia eutropha, which is a representative PHA-producing bacterium with a class I PHA synthase. This is the first report for the cloning of the PHA synthase gene from Massilia species. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Infectivity of pre-seroconversion donations: an analysis of lookback exercises in The Netherlands, 2000-2006.

PubMed

Lieshout-Krikke, R W; van 't Ende, E A; Slot, E; Karomi, S; Kivit, R M H; Zaaijer, H L

2012-04-01

Blood can be infectious if it is donated shortly before infection with hepatitis B virus (HBV), hepatitis C virus (HCV) or human immunodeficiency virus (HIV) becomes detectable. Lookback exercises may detect infection in recipients of pre-seroconversion donations. This study provides an analysis of the Dutch lookback exercises in the years 2000 through 2006. All lookback procedures, triggered by 50 repeat donors seroconverting for HBV (n=32), HCV (n=3), HIV (n=14) and HBV + HIV (n=1), were analysed. Recipients and archived samples of the 96 implicated donations were tested. For 76 donations, a stored sample was available for HBV, HCV, or HIV PCR testing, revealing two HBV-DNA-positive pre-seroconversion donations. Ninety-three lookback procedures were initiated, to which 91 of 93 hospitals responded. In 87 of 91 cases, the implicated blood product had been administered. In 39 of 87 cases, the recipient was tested, revealing one HIV and two HBV infections. The HIV infection was considered pre-existent. The two HBV-positive patients received components from the donation of which the repository sample tested positive for HBV-DNA. Components of the second HBV-positive pre-seroconversion donation had not been administered. Among 39 recipients of pre-seroconversion donations, 2 (5%) were found HBV infected by transfusion. The labour-intensive lookback procedures did not reveal any conclusive transmissions additional to the infections detected by PCR testing of repository pre-seroconversion samples. © 2011 The Author(s). Vox Sanguinis © 2011 International Society of Blood Transfusion.

THE {sup 12}C/{sup 13}C RATIO ON TITAN FROM CASSINI INMS MEASUREMENTS AND IMPLICATIONS FOR THE EVOLUTION OF METHANE

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mandt, Kathleen E.; Waite, J. Hunter; Teolis, Benjamin

2012-04-20

We have re-evaluated the Cassini Ion Neutral Mass Spectrometer (INMS) {sup 12}C/{sup 13}C ratios in the upper atmosphere of Titan based on new calibration sensitivities and an improved model for the NH{sub 3} background in the {sup 13}CH{sub 4} mass channel. The INMS measurements extrapolated to the surface give a {sup 12}C/{sup 13}C in CH{sub 4} of 88.5 {+-} 1.4. We compare the results to a revised ratio of 91.1 {+-} 1.4 provided by the Huygens Gas Chromatograph Mass Spectrometer and 86.5 {+-} 7.9 provided by the Cassini Infrared Spectrometer and determine implications of the revised ratios for the evolutionmore » of methane in Titan's atmosphere. Because the measured {sup 12}C/{sup 13}C is within the probable range of primordial values, we can only determine an upper boundary for the length of time since methane began outgassing from the interior, assuming that outgassing of methane (e.g., cryovolcanic activity) has been continuous ever since. We find that three factors play a crucial role in this timescale: (1) the escape rate of methane, (2) the difference between the current and initial ratios and the rate of methane, and (3) production or resupply due to cryovolcanic activity. We estimate an upper limit for the outgassing timescale of 470 Myr. This duration can be extended to 940 Myr if production rates are large enough to counteract the fractionation due to escape and photochemistry. There is no lower limit to the timescale because the current ratios are within the range of possible primordial values.« less

Creep, creep-rupture tests of Al-surface-alloyed T91 steel in liquid lead bismuth at 500 and 550 °C

NASA Astrophysics Data System (ADS)

Weisenburger, A.; Jianu, A.; An, W.; Fetzer, R.; Del Giacco, Mattia; Heinzel, A.; Müller, G.; Markov, V. G.; Kasthanov, A. D.

2012-12-01

Surface layers made of FeCrAl alloys on T91 steel have shown their capability as corrosion protection barriers in lead bismuth. Pulsed electron beam treatment improves the density and more over the adherence of such layers. After the treatment of previously deposited coatings a surface graded material is achieved with a metallic bonded interface. Creep-rupture tests of T91 in lead-alloy at 550 °C reveal significant reduced creep strength of non-modified T91 test specimens. Oxide scales protecting the steels from attacks of the liquid metal will crack at a certain strain leading to a direct contact between the steel and the liquid metal. The negative influence of the lead-alloy on the creep behavior of non-modified T91 is stress dependent, but below a threshold stress value of 120 MPa at 550 °C this influence becomes almost negligible. At 500 °C and stress values of 200 MPa and 220 MPa the creep rates are comparable between them and significantly lower than creep rates at 180 MPa of original T91 in air at 550 °C. No signs of LBE influence are detected. The surface modified specimens tested at high stress levels instead had creep-rupture times similar to T91 (original state) tested in air. The thin oxide layers formed on the surface modified steel samples are less susceptible to crack formation and therefore to lead-alloy enhanced creep.

Carbon recombination lines as a diagnostic of photodissociation regions

NASA Technical Reports Server (NTRS)

Natta, A.; Walmsley, C. M.; Tielens, A. G. G. M.

1994-01-01

We have observed the C91 alpha radio recombination line toward the Orion H II region. This narrow (approximately 3-5 km per sec full width at half maximum (FWHM)) line is spatially very extended (approximately 8 arcmin or 1 pc). These charateristics compare well with the observed characteristics of the C II fine structure line at 158 microns. Thus, the C91 alpha line originates in the predominantly neutral photodissociation regions separating the H II region from the molecular cloud. We have developed theoretical models for the C II radio recombination lines from photodissociation regions. The results show that the I(C91 alpha)/I(C158) intensity ratio is a sensitive function of the temperature and density of the emitting gas. We have also extended theoretical models for photodissociation regions to include the C II recombination lines. Comparison with these models show that, in the central portion of the Orion region, the C91 alpha line originates in dense (10(exp 6) per cu cm), warm (500-1000 K) gas. Even at large projected distances (approximately 1 pc), the inferred density is still high (10(exp 5) per cu cm) and implies extremely high thermal pressures. As in the case of the (C II) 158 microns line, the large extent of the C91 alpha line shows that (FUV) photons can penetrate to large distances from the illuminating source. The decline of the intensity of the incident radiation field with distance from Theta(sup 1) C seems to be dominated by geometrical dilution, rather than dust extinction. Finally, we have used our models to calculate the intensity of the 9850 A recombination line of C II. The physical conditions inferred from this line are in good agreement with those determined from the radio recombination and the far-infrared fine-structure lines. We show that the ratio of the 9850 A to the C91 alpha lines is a very good probe of very high density clumps.

9 CFR 91.3 - General export requirements.

Code of Federal Regulations, 2014 CFR

2014-01-01

.... (c) Testing. All samples for tests required by §§ 91.5 through 91.13 for exportation of animals under... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false General export requirements. 91.3 Section 91.3 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF...

9 CFR 91.3 - General export requirements.

Code of Federal Regulations, 2012 CFR

2012-01-01

.... (c) Testing. All samples for tests required by §§ 91.5 through 91.13 for exportation of animals under... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false General export requirements. 91.3 Section 91.3 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF...

9 CFR 91.3 - General export requirements.

Code of Federal Regulations, 2013 CFR

2013-01-01

.... (c) Testing. All samples for tests required by §§ 91.5 through 91.13 for exportation of animals under... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false General export requirements. 91.3 Section 91.3 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF...

9 CFR 91.3 - General export requirements.

Code of Federal Regulations, 2011 CFR

2011-01-01

.... (c) Testing. All samples for tests required by §§ 91.5 through 91.13 for exportation of animals under... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false General export requirements. 91.3 Section 91.3 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF...

Significant Role for Microbial Autotrophy in the Sequestration of Soil Carbon

PubMed Central

Yuan, Hongzhao; Ge, Tida; Chen, Caiyan; O'Donnell, Anthony G.

2012-01-01

Soils were incubated for 80 days in a continuously labeled 14CO2 atmosphere to measure the amount of labeled C incorporated into the microbial biomass. Microbial assimilation of 14C differed between soils and accounted for 0.12% to 0.59% of soil organic carbon (SOC). Assuming a terrestrial area of 1.4 × 108 km2, this represents a potential global sequestration of 0.6 to 4.9 Pg C year−1. Estimated global C sequestration rates suggest a “missing sink” for carbon of between 2 and 3 Pg C year−1. To determine whether 14CO2 incorporation was mediated by autotrophic microorganisms, the diversity and abundance of CO2-fixing bacteria and algae were investigated using clone library sequencing, terminal restriction fragment length polymorphism (T-RFLP), and quantitative PCR (qPCR) of the ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) gene (cbbL). Phylogenetic analysis showed that the dominant cbbL-containing bacteria were Azospirillum lipoferum, Rhodopseudomonas palustris, Bradyrhizobium japonicum, Ralstonia eutropha, and cbbL-containing chromophytic algae of the genera Xanthophyta and Bacillariophyta. Multivariate analyses of T-RFLP profiles revealed significant differences in cbbL-containing microbial communities between soils. Differences in cbbL gene diversity were shown to be correlated with differences in SOC content. Bacterial and algal cbbL gene abundances were between 106 and 108 and 103 to 105 copies g−1 soil, respectively. Bacterial cbbL abundance was shown to be positively correlated with RubisCO activity (r = 0.853; P < 0.05), and both cbbL abundance and RubisCO activity were significantly related to the synthesis rates of [14C]SOC (r = 0.967 and 0.946, respectively; P < 0.01). These data offer new insights into the importance of microbial autotrophy in terrestrial C cycling. PMID:22286999

Interaction of Gas Phase Oxalic Acid with Ammonia and its Atmospheric Implications

DOE Office of Scientific and Technical Information (OSTI.GOV)

Peng, Xiu-Qiu; Liu, Yi-Rong; Huang, Teng

Oxalic acid is believed to play an important role in the formation and growth of atmospheric organic aerosols. However, as a common organic acid, the understanding of the larger clusters formed by gas phase oxalic acid with multiple ammonia molecules is incomplete. In this work, the structural characteristics and thermodynamics of oxalic acid clusters with up to six ammonia molecules have been investigated at the PW91PW91/6-311++G(3df,3pd) level of theory. We found that oxalic acid forms relatively stable clusters with ammonia molecules, and that ionization events play a key role. The analyses of the thermodynamics and atmospheric relevance indicate that themore » heterodimer (H2C2O4)(NH3) shows an obvious relative concentration in the atmosphere, and thus likely participates in new particle formation. However, with increasing number of ammonia molecules, the concentration of clusters decreases gradually. Additionally, clusters of oxalic acid with ammonia molecules are predicted to form favorably in low temperature conditions and show high Rayleigh scattering intensities.« less

Prayer and forgiveness: Beyond relationship quality and extension to marriage.

PubMed

Fincham, Frank D; May, Ross W

2017-09-01

The majority of the world population profess religious/spiritual beliefs and prayer is a form of spiritual activity common across numerous religious/spiritual belief systems. Three studies therefore examined the role of prayer in romantic relationships. Study 1 (n = 91) showed that prayer for a dating partner predicted lower aggressive tendencies and greater forgiveness of partner transgressions, independently of relationship closeness. Study 2 (n = 89 married couples) is among the first to examine the prayer-forgiveness association using dyadic data. Controlling for relationship satisfaction in the actor-partner interdependence model, prayer for the spouse predicted own forgiveness but not partner's reports of their own forgiveness. To obviate the problem of obtaining all the data from the same reporter, Study 3 (n = 91 married couples) used partner reports of the spouse's forgiveness in an actor-partner interdependence model analysis. Controlling for religiosity, the results showed that prayer for the partner predicted partner reports of the prayer's forgiveness. The implications of these results are then discussed. (PsycINFO Database Record (c) 2017 APA, all rights reserved).

Department of the Army Supply Bulletin, Army Medical Department Supply Information, SB8-75-S10

DTIC Science & Technology

2001-10-20

12 BTL FLUORESCEIN NA OPTH STRIPS 1 MG 300s 6505-01-159-1493 91C $125.24 1 PG GUAIFENESIN / DEXTRAMETHORAPHAN COUGH SYRUP 4...oz 6505-01-318-1565 91B $1.00 12 BTL GUAIFENESIN EXTENDED RELEASE TABLETS 600MG 100s 6505-01-238-9443 91C $3.89 3 BTL HEMORRHOIDAL

40 CFR 91.409 - Engine dynamometer test run.

Code of Federal Regulations, 2014 CFR

2014-07-01

... all pre-test data specified in § 91.405(c). (4) Start the test cycle (see § 91.410) within 10 minutes... 40 Protection of Environment 20 2014-07-01 2013-07-01 true Engine dynamometer test run. 91.409... (CONTINUED) CONTROL OF EMISSIONS FROM MARINE SPARK-IGNITION ENGINES Gaseous Exhaust Test Procedures § 91.409...

40 CFR 91.409 - Engine dynamometer test run.

Code of Federal Regulations, 2010 CFR

2010-07-01

... all pre-test data specified in § 91.405(c). (4) Start the test cycle (see § 91.410) within 10 minutes... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Engine dynamometer test run. 91.409... (CONTINUED) CONTROL OF EMISSIONS FROM MARINE SPARK-IGNITION ENGINES Gaseous Exhaust Test Procedures § 91.409...

40 CFR 91.409 - Engine dynamometer test run.

Code of Federal Regulations, 2013 CFR

2013-07-01

... all pre-test data specified in § 91.405(c). (4) Start the test cycle (see § 91.410) within 10 minutes... 40 Protection of Environment 21 2013-07-01 2013-07-01 false Engine dynamometer test run. 91.409... (CONTINUED) CONTROL OF EMISSIONS FROM MARINE SPARK-IGNITION ENGINES Gaseous Exhaust Test Procedures § 91.409...

40 CFR 91.409 - Engine dynamometer test run.

Code of Federal Regulations, 2012 CFR

2012-07-01

... all pre-test data specified in § 91.405(c). (4) Start the test cycle (see § 91.410) within 10 minutes... 40 Protection of Environment 21 2012-07-01 2012-07-01 false Engine dynamometer test run. 91.409... (CONTINUED) CONTROL OF EMISSIONS FROM MARINE SPARK-IGNITION ENGINES Gaseous Exhaust Test Procedures § 91.409...

40 CFR 91.409 - Engine dynamometer test run.

Code of Federal Regulations, 2011 CFR

2011-07-01

... all pre-test data specified in § 91.405(c). (4) Start the test cycle (see § 91.410) within 10 minutes... 40 Protection of Environment 20 2011-07-01 2011-07-01 false Engine dynamometer test run. 91.409... (CONTINUED) CONTROL OF EMISSIONS FROM MARINE SPARK-IGNITION ENGINES Gaseous Exhaust Test Procedures § 91.409...

High-Resolution Structure and Mechanism of an F/V-Hybrid Rotor Ring in a Na+-coupled ATP Synthase

PubMed Central

Matthies, Doreen; Zhou, Wenchang; Klyszejko, Adriana L.; Anselmi, Claudio; Yildiz, Özkan; Brandt, Karsten; Müller, Volker; Faraldo-Gómez, José D.; Meier, Thomas

2014-01-01

All rotary ATPases catalyze the interconversion of ATP and ADP-Pi through a mechanism that is coupled to the transmembrane flow of H+ or Na+. Physiologically, however, F/A-type enzymes specialize in ATP synthesis driven by downhill ion diffusion, while eukaryotic V-type ATPases function as ion pumps. To begin to rationalize the molecular basis for this functional differentiation, we solved the crystal structure of the Na+-driven membrane rotor of the Acetobacterium woodii ATP synthase, at 2.1 Å resolution. Unlike known structures, this rotor ring is a 9:1 heteromer of F- and V-type c-subunits, and therefore features a hybrid configuration of ion-binding sites along its circumference. Molecular and kinetic simulations are used to dissect the mechanisms of Na+ recognition and rotation of this c-ring, and to explain the functional implications of the V-type c-subunit. These structural and mechanistic insights indicate an evolutionary path between synthases and pumps involving adaptations in the rotor ring. PMID:25381992

40 CFR 91.123 - Denial, revocation of certificate of conformity.

Code of Federal Regulations, 2010 CFR

2010-07-01

....203(f), 91.206(d), 91.208(c) or 91.209(g), the Administrator may void such certificate ab initio. (d) When the Administrator denies, revokes, or voids ab initio a certificate of conformity, the engine... makes the certificate void ab initio. ...

40 CFR 91.123 - Denial, revocation of certificate of conformity.

Code of Federal Regulations, 2013 CFR

2013-07-01

....203(f), 91.206(d), 91.208(c) or 91.209(g), the Administrator may void such certificate ab initio. (d) When the Administrator denies, revokes, or voids ab initio a certificate of conformity, the engine... makes the certificate void ab initio. ...

40 CFR 91.123 - Denial, revocation of certificate of conformity.

Code of Federal Regulations, 2011 CFR

2011-07-01

....203(f), 91.206(d), 91.208(c) or 91.209(g), the Administrator may void such certificate ab initio. (d) When the Administrator denies, revokes, or voids ab initio a certificate of conformity, the engine... makes the certificate void ab initio. ...

40 CFR 91.123 - Denial, revocation of certificate of conformity.

Code of Federal Regulations, 2012 CFR

2012-07-01

....203(f), 91.206(d), 91.208(c) or 91.209(g), the Administrator may void such certificate ab initio. (d) When the Administrator denies, revokes, or voids ab initio a certificate of conformity, the engine... makes the certificate void ab initio. ...

40 CFR 91.123 - Denial, revocation of certificate of conformity.

Code of Federal Regulations, 2014 CFR

2014-07-01

....203(f), 91.206(d), 91.208(c) or 91.209(g), the Administrator may void such certificate ab initio. (d) When the Administrator denies, revokes, or voids ab initio a certificate of conformity, the engine... makes the certificate void ab initio. ...

[Recombinant OspC identification and antigenicity detection from Borrelia burgdorferi PD91 in China].

PubMed

Chen, Jian; Wan, Kang-Lin

2003-10-01

To recombine OspC gene from Borrelia burgdorferi PD91 of China and expressed it in E. coli for early diagnosis of Lyme disease. The OspC gene was amplified from the genome of Borrelia burgdorferi PD91 strain by polymerase chain reaction and recombined with plasmid PET-11D. The recombinant plasmid PET-11D-OspC was identified with PCR, restriction endonuclease analysis and sequencing. The antigenicity was verified with Western Blot. OspC gene was cloned correctly into vector PET-11D. The resultant sequence was definitely different from the published sequence. The recombinant OspC seemed to have had strong antigenicity. The findings laid basis for the studies on early diagnosis of Lyme disease.

14 CFR 91.517 - Passenger information.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 14 Aeronautics and Space 2 2010-01-01 2010-01-01 false Passenger information. 91.517 Section 91... Multiengine Airplanes and Fractional Ownership Program Aircraft § 91.517 Passenger information. (a) Except as... belts and when smoking is prohibited. (c) If passenger information signs are installed, no passenger or...

24 CFR 91.315 - Strategic plan.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 24 Housing and Urban Development 1 2011-04-01 2011-04-01 false Strategic plan. 91.315 Section 91... CONSOLIDATED SUBMISSIONS FOR COMMUNITY PLANNING AND DEVELOPMENT PROGRAMS State Governments; Contents of Consolidated Plan § 91.315 Strategic plan. (a) General. For the categories described in paragraphs (b), (c), (d...

Bimodality emerges from transport model calculations of heavy ion collisions at intermediate energy

NASA Astrophysics Data System (ADS)

Mallik, S.; Das Gupta, S.; Chaudhuri, G.

2016-04-01

This work is a continuation of our effort [S. Mallik, S. Das Gupta, and G. Chaudhuri, Phys. Rev. C 91, 034616 (2015)], 10.1103/PhysRevC.91.034616 to examine if signatures of a phase transition can be extracted from transport model calculations of heavy ion collisions at intermediate energy. A signature of first-order phase transition is the appearance of a bimodal distribution in Pm(k ) in finite systems. Here Pm(k ) is the probability that the maximum of the multiplicity distribution occurs at mass number k . Using a well-known model for event generation [Botzmann-Uehling-Uhlenbeck (BUU) plus fluctuation], we study two cases of central collision: mass 40 on mass 40 and mass 120 on mass 120. Bimodality is seen in both the cases. The results are quite similar to those obtained in statistical model calculations. An intriguing feature is seen. We observe that at the energy where bimodality occurs, other phase-transition-like signatures appear. There are breaks in certain first-order derivatives. We then examine if such breaks appear in standard BUU calculations without fluctuations. They do. The implication is interesting. If first-order phase transition occurs, it may be possible to recognize that from ordinary BUU calculations. Probably the reason this has not been seen already is because this aspect was not investigated before.

Microstructure characterization and charpy toughness of P91 weldment for as-welded, post-weld heat treatment and normalizing & tempering heat treatment

NASA Astrophysics Data System (ADS)

Pandey, Chandan; Mahapatra, M. M.; Kumar, Pradeep; Giri, A.

2017-09-01

The effect of weld groove design and heat treatment on microstructure evolution and Charpy toughness of P91 pipe weldments was studied. The P91 pipe weldments were subjected to subcritical post weld heat treatment (760 °C-2 h) and normalizing/tempering conditions (normalized-1040 °C/40 min, air cooled; tempered 760 °C/2 h, air cooled) were employed. The influence of subsequent PWHT and N&T treatment on the microstructure of various zone of P91 pipe weldments were also investigated. The present investigation also described the effect of PWHT and N&T treatment on hardness, grain size, precipitate size, inter-particle spacing and fraction area of precipitates present in each zone of P91 pipe weldments. The result indicated great impact of heat treatment on the Charpy toughness and microstructure evolution of P91 weldments. The N&T treatment was found to be more effective heat treatment compared to subsequent PWHT. Charpy toughness value was found to be higher for narrow-groove design as compared to conventional V-groove design.

11 CFR 100.91 - Recounts.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 11 Federal Elections 1 2011-01-01 2011-01-01 false Recounts. 100.91 Section 100.91 Federal Elections FEDERAL ELECTION COMMISSION GENERAL SCOPE AND DEFINITIONS (2 U.S.C. 431) Exceptions to Contributions § 100.91 Recounts. A gift, subscription, loan, advance, or deposit of money or anything of value...

11 CFR 100.91 - Recounts.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 11 Federal Elections 1 2010-01-01 2010-01-01 false Recounts. 100.91 Section 100.91 Federal Elections FEDERAL ELECTION COMMISSION GENERAL SCOPE AND DEFINITIONS (2 U.S.C. 431) Exceptions to Contributions § 100.91 Recounts. A gift, subscription, loan, advance, or deposit of money or anything of value...

Rubredoxin-related Maturation Factor Guarantees Metal Cofactor Integrity during Aerobic Biosynthesis of Membrane-bound [NiFe] Hydrogenase*

PubMed Central

Fritsch, Johannes; Siebert, Elisabeth; Priebe, Jacqueline; Zebger, Ingo; Lendzian, Friedhelm; Teutloff, Christian; Friedrich, Bärbel; Lenz, Oliver

2014-01-01

The membrane-bound [NiFe] hydrogenase (MBH) supports growth of Ralstonia eutropha H16 with H2 as the sole energy source. The enzyme undergoes a complex biosynthesis process that proceeds during cell growth even at ambient O2 levels and involves 14 specific maturation proteins. One of these is a rubredoxin-like protein, which is essential for biosynthesis of active MBH at high oxygen concentrations but dispensable under microaerobic growth conditions. To obtain insights into the function of HoxR, we investigated the MBH protein purified from the cytoplasmic membrane of hoxR mutant cells. Compared with wild-type MBH, the mutant enzyme displayed severely decreased hydrogenase activity. Electron paramagnetic resonance and infrared spectroscopic analyses revealed features resembling those of O2-sensitive [NiFe] hydrogenases and/or oxidatively damaged protein. The catalytic center resided partially in an inactive Niu-A-like state, and the electron transfer chain consisting of three different Fe-S clusters showed marked alterations compared with wild-type enzyme. Purification of HoxR protein from its original host, R. eutropha, revealed only low protein amounts. Therefore, recombinant HoxR protein was isolated from Escherichia coli. Unlike common rubredoxins, the HoxR protein was colorless, rather unstable, and essentially metal-free. Conversion of the atypical iron-binding motif into a canonical one through genetic engineering led to a stable reddish rubredoxin. Remarkably, the modified HoxR protein did not support MBH-dependent growth at high O2. Analysis of MBH-associated protein complexes points toward a specific interaction of HoxR with the Fe-S cluster-bearing small subunit. This supports the previously made notion that HoxR avoids oxidative damage of the metal centers of the MBH, in particular the unprecedented Cys6[4Fe-3S] cluster. PMID:24448806

rRNA and Poly-β-Hydroxybutyrate Dynamics in Bioreactors Subjected to Feast and Famine Cycles

PubMed Central

Frigon, Dominic; Muyzer, Gerard; van Loosdrecht, Mark; Raskin, Lutgarde

2006-01-01

Feast and famine cycles are common in activated sludge wastewater treatment systems, and they select for bacteria that accumulate storage compounds, such as poly-β-hydroxybutyrate (PHB). Previous studies have shown that variations in influent substrate concentrations force bacteria to accumulate high levels of rRNA compared to the levels in bacteria grown in chemostats. Therefore, it can be hypothesized that bacteria accumulate more rRNA when they are subjected to feast and famine cycles. However, PHB-accumulating bacteria can form biomass (grow) throughout a feast and famine cycle and thus have a lower peak biomass formation rate during the cycle. Consequently, PHB-accumulating bacteria may accumulate less rRNA when they are subjected to feast and famine cycles than bacteria that are not capable of PHB accumulation. These hypotheses were tested with Wautersia eutropha H16 (wild type) and W. eutropha PHB-4 (a mutant not capable of accumulating PHB) grown in chemostat and semibatch reactors. For both strains, the cellular RNA level was higher when the organism was grown in semibatch reactors than when it was grown in chemostats, and the specific biomass formation rates during the feast phase were linearly related to the cellular RNA levels for cultures. Although the two strains exhibited maximum uptake rates when they were grown in semibatch reactors, the wild-type strain responded much more rapidly to the addition of fresh medium than the mutant responded. Furthermore, the chemostat-grown mutant culture was unable to exhibit maximum substrate uptake rates when it was subjected to pulse-wise addition of fresh medium. These data show that the ability to accumulate PHB does not prevent bacteria from accumulating high levels of rRNA when they are subjected to feast and famine cycles. Our results also demonstrate that the ability to accumulate PHB makes the bacteria more responsive to sudden increases in substrate concentrations, which explains their ecological advantage. PMID:16597926

rRNA and poly-beta-hydroxybutyrate dynamics in bioreactors subjected to feast and famine cycles.

PubMed

Frigon, Dominic; Muyzer, Gerard; van Loosdrecht, Mark; Raskin, Lutgarde

2006-04-01

Feast and famine cycles are common in activated sludge wastewater treatment systems, and they select for bacteria that accumulate storage compounds, such as poly-beta-hydroxybutyrate (PHB). Previous studies have shown that variations in influent substrate concentrations force bacteria to accumulate high levels of rRNA compared to the levels in bacteria grown in chemostats. Therefore, it can be hypothesized that bacteria accumulate more rRNA when they are subjected to feast and famine cycles. However, PHB-accumulating bacteria can form biomass (grow) throughout a feast and famine cycle and thus have a lower peak biomass formation rate during the cycle. Consequently, PHB-accumulating bacteria may accumulate less rRNA when they are subjected to feast and famine cycles than bacteria that are not capable of PHB accumulation. These hypotheses were tested with Wautersia eutropha H16 (wild type) and W. eutropha PHB-4 (a mutant not capable of accumulating PHB) grown in chemostat and semibatch reactors. For both strains, the cellular RNA level was higher when the organism was grown in semibatch reactors than when it was grown in chemostats, and the specific biomass formation rates during the feast phase were linearly related to the cellular RNA levels for cultures. Although the two strains exhibited maximum uptake rates when they were grown in semibatch reactors, the wild-type strain responded much more rapidly to the addition of fresh medium than the mutant responded. Furthermore, the chemostat-grown mutant culture was unable to exhibit maximum substrate uptake rates when it was subjected to pulse-wise addition of fresh medium. These data show that the ability to accumulate PHB does not prevent bacteria from accumulating high levels of rRNA when they are subjected to feast and famine cycles. Our results also demonstrate that the ability to accumulate PHB makes the bacteria more responsive to sudden increases in substrate concentrations, which explains their ecological advantage.

FIELD MEASUREMENT TECHNOLOGY FOR MERCURY IN SOIL AND SEDIMENT OHIO LUMEX'S RA-915+/RP-91C MERCURY ANALYZER

EPA Science Inventory

Ohio Lumex's RA915+/91 C mercury analyzer was demonstrated under the U.S. Environmental Protection Agency Superfund Innovative Technology Evaluation Program in May 2003, at the Oak Ridge National Laboratory (ORNL) in Oak Ridge, Tennessee. The purpose of the Demonstration was to c...

70. TILE WORKS FROM SOUTH, C. 1916. SL/BCHS, PHOTOGRAPH 8N91. ...

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

70. TILE WORKS FROM SOUTH, C. 1916. SL/BCHS, PHOTOGRAPH 8-N-91. - Moravian Pottery & Tile Works, Southwest side of State Route 313 (Swamp Road), Northwest of East Court Street, Doylestown, Bucks County, PA

40 CFR 91.314 - Analyzer accuracy and specifications.

Code of Federal Regulations, 2012 CFR

2012-07-01

... to § 91.325 and § 91.409. (c) Emission measurement accuracy—bag sampling. (1) Good engineering... deflection should generally not be used. (2) Some high resolution read-out systems, such as computers, data...

40 CFR 91.314 - Analyzer accuracy and specifications.

Code of Federal Regulations, 2013 CFR

2013-07-01

... to § 91.325 and § 91.409. (c) Emission measurement accuracy—bag sampling. (1) Good engineering... deflection should generally not be used. (2) Some high resolution read-out systems, such as computers, data...

40 CFR 91.314 - Analyzer accuracy and specifications.

Code of Federal Regulations, 2011 CFR

2011-07-01

... to § 91.325 and § 91.409. (c) Emission measurement accuracy—bag sampling. (1) Good engineering... deflection should generally not be used. (2) Some high resolution read-out systems, such as computers, data...

Effect of SiC Nanoparticles on Hot Deformation Behavior and Processing Maps of Magnesium Alloy AZ91

PubMed Central

Nie, Kaibo; Kang, Xinkai; Deng, Kunkun; Wang, Ting; Guo, Yachao; Wang, Hongxia

2018-01-01

The hot deformation behavior and processing characteristics of AZ91 alloy and nano-SiCp/AZ91 composite were compared at temperature ranges of 523 K–673 K and strain rates of 0.001–1 s−1. Positive impact of SiC nanoparticles on pinning grain boundaries and inhibiting grain growth was not obvious when deformation temperature was below 623 K, but was remarkable when the temperature was above 623 K. By comparing compressive stress-strain curves of AZ91 alloy and nano-SiCp/AZ91 composites, the addition of nanoparticles could improve the deformation ability of a matrix alloy under high-temperature conditions. There was no essential difference of deformation mechanism between AZ91 alloy and the composite, but hot deformation activation energy of the composite was significantly lower than that of the AZ91 alloy. The AZ91 alloy and the composite had the same workability region of 600 K–673 K and 0.001–1 s−1, while instability region for the composite was reduced compared with that of AZ91 alloy at high temperature. PMID:29389888

G protein-coupled receptor 91 signaling in diabetic retinopathy and hypoxic retinal diseases.

PubMed

Hu, Jianyan; Li, Tingting; Du, Xinhua; Wu, Qiang; Le, Yun-Zheng

2017-10-01

G protein-coupled receptor 91 (GPR91) is a succinate-specific receptor and activation of GPR91 could initiate a complex signal transduction cascade and upregulate inflammatory and pro-angiogenic cytokines. In the retina, GPR91 is predominately expressed in ganglion cells, a major cellular entity involved in the pathogenesis of diabetic retinopathy (DR) and other hypoxic retinal diseases. During the development of DR and retinopathy of prematurity (ROP), chronic hypoxia causes an increase in the levels of local succinate. Succinate-mediated GPR91 activation upregulates vascular endothelial growth factor (VEGF) through ERK1/2-C/EBP β (c-Fos) and/or ERK1/2-COX-2/PGE2 signaling pathways, which in turn, leads to the breakdown of blood-retina barriers in these disorders. In this review, we will have a brief introduction of GPR91 and its biological functions and a more detailed discussion about the role and mechanisms of GPR91 in DR and ROP. A better understanding of GPR91 regulation may be of great significance in identifying new biomarkers and drug targets for the prediction and treatment of DR, ROP, and hypoxic retinal diseases. Copyright © 2017 Elsevier Ltd. All rights reserved.

Effect of ferrocene-substituted porphyrin RL-91 on Candida albicans biofilm formation.

PubMed

Lippert, Rainer; Vojnovic, Sandra; Mitrovic, Aleksandra; Jux, Norbert; Ivanović-Burmazović, Ivana; Vasiljevic, Branka; Stankovic, Nada

2014-08-01

Ferrocene-substituted porphyrin RL-91 exhibits antifungal activity against opportune human pathogen Candida albicans. RL-91 efficiently inhibits growth of both planktonic C. albicans cells and cells within biofilms without photoactivation. The minimal inhibitory concentration for plankton form (PMIC) was established to be 100 μg/mL and the same concentration killed 80% of sessile cells in the mature biofilm (SMIC80). Furthermore PMIC of RL-91 efficiently prevents C. albicans biofilm formation. RL-91 is cytotoxic for human fibroblasts in vitro in concentration of 10 μg/mL, however it does not cause hemolysis in concentrations of up to 50 μg/mL. These findings open possibility for application of RL-91 as an antifungal agent for external antibiofilm treatment of medical devices as well as a scaffold for further development of porphyrin based systemic antifungals. Copyright © 2014 Elsevier Ltd. All rights reserved.

Chemical stability of extemporaneously prepared Lorazepam suspension at two temperatures.

PubMed

Lee, Wan-Man Ellaria; Lugo, Ralph A; Rusho, William J; Mackay, Mark; Sweeley, John

2004-10-01

The objective of this study was to determine the chemical stability of extemporaneously prepared lorazepam suspension (1 mg/mL) stored at two temperatures (4°C and 22°C) for 3 months. Lorazepam tablets marketed by two manufacturers (Mylan Pharmaceuticals and Watson Laboratories) were used to extemporaneously formulate two independently prepared suspensions. Each suspension was prepared using sterile water, Ora-Plus(®) and Ora-Sweet(®) to achieve a final concentration of 1 mg/mL. The two brands of tablets required different volumes of vehicles to prepare a pharmaceutically optimal suspension. The suspensions were stored in amber glass bottles at 4°C and 22°C for 91 days. Samples were analyzed by high performance liquid chromatography at baseline and on days 2, 3, 7, 14, 21, 28, 42, 63, and 91. The suspensions were considered stable if the mean lorazepam concentration remained greater than 90% of the initial concentration.The chemical stabilities of these two extemporaneously prepared lorazepam suspensions were comparable throughout the study. Both lorazepam suspensions were stable for 63 days when stored at 4°C or 22°C, and both were stable for 91 days when refrigerated at 4°C. When stored at room temperature, the suspension prepared from the Watson tablet retained 88.9 ± 1.4% of the initial concentration on day 91 and was therefore considered unstable, while the suspension prepared from the Mylan tablet was stable for the entire 91-day study.

77 FR 28668 - Technical Standard Order (TSO)-C91a, Emergency Locator Transmitters (ELTs)

Federal Register 2010, 2011, 2012, 2013, 2014

2012-05-15

... installed in new production airplanes for over the past 5-years. The FAA reiterates in this final notice... questions. The first question regarded permissibility of minor and major changes to existing TSO-C91a... meet the minimum performance standards contained in TSO-C126a. Cobham's second question requested...

Nutritional environment influences transcription of a pantocin A biosynthesis gene in Pantoea vagans strain C9-1

USDA-ARS?s Scientific Manuscript database

The histidine-reversible antibiotic pantocin A is produced by Pantoea vagans strain C9-1 and Pantoea agglomerans strains Eh252, EH318 and P10c and contributes significantly to efficacy of biological control of fire blight of pear and apple flowers caused by Erwinia amylovora. Antibiosis by pantocin ...

Microstructural evolution of neutron-irradiated T91 and NF616 to ~4.3 dpa at 469 °C

DOE PAGES

Tan, Lizhen; Kim, B. K.; Yang, Ying; ...

2017-05-30

Ferritic-martensitic steels such as T91 and NF616 are candidate materials for several nuclear applications. Here, this study evaluates radiation resistance of T91 and NF616 by examining their microstructural evolutions and hardening after the samples were irradiated in the Advanced Test Reactor to ~4.3 displacements per atom (dpa) at an as-run temperature of 469 °C. In general, this irradiation did not result in significant difference in the radiation-induced microstructures between the two steels. Compared to NF616, T91 had a higher number density of dislocation loops and a lower level of radiation-induced segregation, together with a slightly higher radiation-hardening. Unlike dislocation loopsmore » developed in both steels, radiation-induced cavities were only observed in T91 but remained small with sub-10 nm sizes. Lastly, other than the relatively stable M 23C 6, a new phase (likely Sigma phase) was observed in T91 and radiation-enhanced MX → Z phase transformation was identified in NF616. Laves phase was not observed in the samples.« less

45 CFR 96.91 - Audit requirement.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 45 Public Welfare 1 2010-10-01 2010-10-01 false Audit requirement. 96.91 Section 96.91 Public... Block Grants § 96.91 Audit requirement. Pursuant to section 1745(b) of the Reconciliation Act (31 U.S.C. 1243 note) an audit is required with respect to the 2-year period beginning on October 1, 1981, and...

14 CFR 91.853 - Final compliance: Civil subsonic airplanes.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 14 Aeronautics and Space 2 2012-01-01 2012-01-01 false Final compliance: Civil subsonic airplanes... Noise Limits § 91.853 Final compliance: Civil subsonic airplanes. Except as provided in § 91.873, after... airplane subject to § 91.801(c) of this subpart, unless that airplane has been shown to comply with Stage 3...

14 CFR 91.853 - Final compliance: Civil subsonic airplanes.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 14 Aeronautics and Space 2 2014-01-01 2014-01-01 false Final compliance: Civil subsonic airplanes... Noise Limits § 91.853 Final compliance: Civil subsonic airplanes. Except as provided in § 91.873, after... airplane subject to § 91.801(c) of this subpart, unless that airplane has been shown to comply with Stage 3...

14 CFR 91.853 - Final compliance: Civil subsonic airplanes.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 14 Aeronautics and Space 2 2011-01-01 2011-01-01 false Final compliance: Civil subsonic airplanes... Noise Limits § 91.853 Final compliance: Civil subsonic airplanes. Except as provided in § 91.873, after... airplane subject to § 91.801(c) of this subpart, unless that airplane has been shown to comply with Stage 3...

14 CFR 91.853 - Final compliance: Civil subsonic airplanes.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 14 Aeronautics and Space 2 2013-01-01 2013-01-01 false Final compliance: Civil subsonic airplanes... Noise Limits § 91.853 Final compliance: Civil subsonic airplanes. Except as provided in § 91.873, after... airplane subject to § 91.801(c) of this subpart, unless that airplane has been shown to comply with Stage 3...

14 CFR 91.853 - Final compliance: Civil subsonic airplanes.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 14 Aeronautics and Space 2 2010-01-01 2010-01-01 false Final compliance: Civil subsonic airplanes... Noise Limits § 91.853 Final compliance: Civil subsonic airplanes. Except as provided in § 91.873, after... airplane subject to § 91.801(c) of this subpart, unless that airplane has been shown to comply with Stage 3...

Creep Strength Behavior of Boron Added P91 Steel and its Weld in the Temperature Range of 600-650°C

NASA Astrophysics Data System (ADS)

Swaminathan, J.; Das, C. R.; Baral, Jayashree; Phaniraj, C.; Ghosh, R. N.; Albert, S. K.; Bhaduri, A. K.

One of the promising ways for mitigation of Type IV cracking — a failure by cracking at the intercritical /fine grained heat affected zone, a life limiting problem in advanced 9-12 Cr ferritic steel weld like that of P91 is through modification of alloy composition by addition of boron. Addition of boron was observed to improve the microstructure at the weld zone and hence the creep strength. In the present work, boron (100 ppm with controlled nitrogen) added P91 steel after normalizing at 1050°C and 1150°C and tempered at 760°C were studied for the creep behavior in the base metal and welded condition in the temperature range of 600-650°C. Creep strength was characterized in terms of stress and temperature dependence of creep rate and rupture time. Weld creep life was reduced compared to the base metal with rupture occurring at the ICHAZ (Type IV crack). However at longer time (at lower stress levels) exposure creep crack moves from weld metal to HAZ (Type II crack). Rupture life was found to superior for the base and weld in the boron containing steel when higher normalizing temperature is used. Estimation of 105 h was attempted based on short term rupture data available and weld strength factors were calculated. Observed values are better for P91BH condition than the values for P91BLcondition as well as those available for P91 in open literature

Proteins with CHADs (Conserved Histidine α-Helical Domains) Are Attached to Polyphosphate Granules In Vivo and Constitute a Novel Family of Polyphosphate-Associated Proteins (Phosins).

PubMed

Tumlirsch, Tony; Jendrossek, Dieter

2017-04-01

On the basis of bioinformatic evidence, we suspected that proteins with a CYTH ( Cy aB th iamine triphosphatase) domain and/or a CHAD ( c onserved h istidine α -helical d omain) motif might represent polyphosphate (polyP) granule-associated proteins. We found no evidence of polyP targeting by proteins with CYTH domains. In contrast, two CHAD motif-containing proteins from Ralstonia eutropha H16 (A0104 and B1017) that were expressed as fusions with enhanced yellow fluorescent protein (eYFP) colocalized with polyP granules. While the expression of B1017 was not detectable, the A0104 protein was specifically identified in an isolated polyP granule fraction by proteome analysis. Moreover, eYFP fusions with the CHAD motif-containing proteins MGMSRV2-1987 from Magnetospirillum gryphiswaldense and PP2307 from Pseudomonas putida also colocalized with polyP granules in a transspecies-specific manner. These data indicated that CHAD-containing proteins are generally attached to polyP granules. Together with the findings from four previously polyP-attached proteins (polyP kinases), the results of this study raised the number of polyP-associated proteins in R. eutropha to six. We suggest designating polyP granule-bound proteins with CHAD motifs as phosins ( pho sphate), analogous to pha sins and oleo sins that are specifically bound to the surface of polyhydroxyalkanoate (PHA) granules in PHA-accumulating bacteria and to oil droplets in oil seed plants, respectively. IMPORTANCE The importance of polyphosphate (polyP) for life is evident from the ubiquitous presence of polyP in all species on earth. In unicellular eukaryotic microorganisms, polyP is located in specific membrane-enclosed organelles, called acidocalcisomes. However, in most prokaryotes, polyP is present as insoluble granules that have been designated previously as volutin granules. Almost nothing is known regarding the macromolecular composition of polyP granules. Particularly, the absence or presence of cellular compounds on the surface of polyP granules has not yet been investigated. In this study, we identified a novel class of proteins that are attached to the surface of polyP granules in three model species of Alphaproteobacteria , Betaproteobacteria , and Gammaproteobacteria These proteins are characterized by the presence of a CHAD ( c onserved h istidine α -helical d omain) motif that functions as a polyP granule-targeting signal. We suggest designating CHAD motif-containing proteins as phosins [analogous to phasins for poly(3-hydroxybutyrate)-associated proteins and to oleosins for oil droplet-associated proteins in oil seed plants]. The expression of phosins in different species confirmed their polyP-targeting function in a transspecies-specific manner. We postulate that polyP granules in prokaryotic species generally have a complex surface structure that consists of one to several polyP kinases and phosin proteins. We suggest differentiating polyP granules from acidocalcisomes by designating them as polyphosphatosomes. Copyright © 2017 American Society for Microbiology.

Prime Contract Awards Alphabetically by Contractor, by State or Country, and Place. Part 7. (Fahey Watts Architects-General Contract and Sales)

DTIC Science & Technology

1989-01-01

0 -m Il 5 ))40 . I: 0 40-4-4 -x000) of I 00-4qN< 66 ."eN *-4NI g - *N .40)-4 .4 U1.4 MOMO t00 m m’ C-eq an .- 4-4 eqI 0 .-44 1 ::-4...991 M (0N lice x W (A 4 4C (5n ( U) n44 1-44e mlt w. < Z 91 (0N eq4 c 0 a >- 0 0u >- >- 00X 1- 0 1-0o 1- cc I-- 0 0 C I M C14 91 -4 W. cc . 5 0D 0 -. 1 .1...001 - 00 IO 0 ( NC1n1- 1-1 C90 1 M 4 I 9(no0 1-- 0 4C 0 . 4 4 ~i 0 - 5 <C 4 ZNNZ 0 91 0 j

Herpes viruses and human papilloma virus in nasal polyposis and controls.

PubMed

Ioannidis, Dimitrios; Lachanas, Vasileios A; Florou, Zoe; Bizakis, John G; Petinaki, Efthymia; Skoulakis, Charalampos E

2015-01-01

Chronic rhinosinusitis with nasal polyps is a multifactorial disease entity with an unclear pathogenesis. Contradictory data exist in the literature on the potential implication of viral elements in adult patients with chronic rhinosinusitis. To compare the prevalence of human herpes viruses (1-6) and Human Papilloma Virus in adult patients with chronic rhinosinusitis with nasal polyps and healthy controls. Viral DNA presence was evaluated by real-time polymerase chain reaction application to nasal polyps specimens from 91 chronic rhinosinusitis with nasal polyps patients and nasal turbinate mucosa from 38 healthy controls. Epstein-Barr virus positivity was higher in nasal polyps (24/91; 26.4%) versus controls (4/38; 10.5%), but the difference did not reach significance (p=0.06). Human herpes virus-6 positivity was lower in nasal polyps (13/91; 14.29%) versus controls (10/38; 26.32%, p=0.13). In chronic rhinosinusitis with nasal polyps group, 1 sample was herpes simplex virus-1-positive (1/91; 1.1%), and another was cytomegalovirus-positive (1/91; 1.1%), versus none in controls. No sample was positive for herpes simplex virus-2, varicella-zoster virus, high-risk-human papilloma viruses (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59) and low-risk-human papilloma viruses (6, 11). Differences in Epstein-Barr virus and human herpes virus-6 positivity among patients with chronic rhinosinusitis with nasal polyps and healthy controls are not statistically significant, weakening the likelihood of their implication in chronic rhinosinusitis with nasal polyps pathogenesis. Copyright © 2015 Associação Brasileira de Otorrinolaringologia e Cirurgia Cérvico-Facial. Published by Elsevier Editora Ltda. All rights reserved.

Stability of extemporaneously compounded dexamethasone in glass and plastic bottles and plastic syringes.

PubMed

Ensom, Mary H H; Décarie, Diane

2014-07-01

Dexamethasone is widely used to treat rheumatic and endocrine disorders and chemotherapy-induced nausea and vomiting. A palatable, alcohol-free liquid formulation, with a suitable concentration to allow reasonable administration volume, is available only via extemporaneous compounding. To evaluate the stability of dexamethasone suspensions in commercially available vehicles (Oral Mix and Oral Mix SF) in various types of containers after storage at 25°C and 4°C for up to 91 days. Dexamethasone suspensions (1 mg/mL) were prepared in Oral Mix and Oral Mix SF and then transferred to amber glass and plastic prescription bottles and plastic oral syringes. Suspensions in all 3 types of containers were stored at 25°C; suspensions in glass and plastic bottles were also stored at 4°C. Samples were collected weekly from each container up to 28 days and then every 2 weeks up to 91 days. The samples were analyzed by a validated, stability-indicating high-performance liquid chromatography - ultraviolet detection method. A suspension was considered stable if it maintained at least 90% of its initial dexamethasone concentration. Changes in colour, taste, odour, precipitation (and ease of resuspension), and pH were used to assess physical compatibility. All suspensions maintained at least 96% of the original concentration for up to 91 days with storage at 25°C or at 4°C. No notable changes in colour, taste, odour, precipitation, or pH were observed over the 91-day period. Dexamethasone suspensions (1 mg/mL) in Oral Mix and Oral Mix SF, stored in amber glass or plastic bottles or plastic syringes at 25°C or in amber glass or plastic bottles at 4°C can be expected to remain stable for up to 91 days.

Buckley ANGB, Colorado. Revised Uniform Summary of Surface Weather Observations (RUSSWO). Parts A-F.

DTIC Science & Technology

1983-09-19

911. 91. 911, 94.1 9.1. 914.1 9,4. 94.1 9c( ;J.t 92.1 92.8 93.1’ 94.? 94.2! 94 11.6 91.6 911.7 91.7i 91.8’ 94.7l 9 .9 95.9 9.19 9.1 & Kc ’ 3 .1 92.1 .F9...1 7, El.....’ (X) l I s ![ .i N. Oh.. N e N.. .4 Nw.~e wil Teq.. * 6 ’ I. N .... ti0P 132F * TeO . 2 11.. , 1 ... .. 4 1.. S.97 • , - -4 a It maml Wd

Neuropsychological implications of selective attentional functioning in psychopathic offenders.

PubMed

Mayer, Andrew R; Kosson, David S; Bedrick, Edward J

2006-09-01

Several core characteristics of the psychopathic personality disorder (i.e., impulsivity, failure to attend to interpersonal cues) suggest that psychopaths suffer from disordered attention. However, there is mixed evidence from the cognitive literature as to whether they exhibit superior or deficient selective attention, which has led to the formation of several distinct theories of attentional functioning in psychopathy. The present experiment investigated participants' abilities to purposely allocate attentional resources on the basis of auditory or visual linguistic information and directly tested both theories of deficient or superior selective attention in psychopathy. Specifically, 91 male inmates at a county jail were presented with either auditory or visual linguistic cues (with and without distractors) that correctly indicated the position of an upcoming visual target in 75% of the trials. The results indicated that psychopaths did not exhibit evidence of superior selective attention in any of the conditions but were generally less efficient in shifting attention on the basis of linguistic cues, especially in regard to auditory information. Implications for understanding psychopaths' cognitive functioning and possible neuropsychological deficits are addressed. ((c) 2006 APA, all rights reserved).

Hydrogen and carbon isotopic composition of volatiles in Nakhla: Implications for weathering on Mars

NASA Technical Reports Server (NTRS)

Watson, L. L.; Epstein, S.; Stolper, E. M.

1992-01-01

Gases were collected at 120, 200, 300, 415, and 600 and 850 C. Hydrogen yields for the 600 and 850 C aliquots were measured separately and then the gases were combined for isotopic analysis. CO2 samples collected at the two lowest temperature steps amounted to less than 0.5 mu mole and were not analyzed isotopically. Excluding the 120 C temperature step, the bulk delta D of the sample was + 187 percent. Delta D values increase from -91 percent in the 120 C step to +518 percent in the 315 to 850 C step. The hydrogen content is greatest in the 120 C step and is roughly constant in the 200, 300, and 415 C aliquots. Between 415 C and 850 C, the yield drops off considerably. From 850 C to 950 C, virtually no H2 and only minor CO2 (less than 1 mu mole) were extracted. Using the isotopic analysis from the 300, 415, 600, and 850 C temperature collections, the bulk delta C-13 sub (PDB) is 0.0 percent. The heaviest component (delta C-13 sub (PDB) of +29 percent) was collected between 300 and 415 C. The release of hydrogen at the low temperatures reported here is consistent with the breakdown of the phases that constitute the alteration product between approx. 250 and 650 C. Although not as high as the present Martian atmosphere, the high delta D values are consistent with a Martian origin for the meteorites in question.

The expression of full length Gp91-phox protein is associated with reduced amphotropic retroviral production.

PubMed

Bellantuono, I; Lashford, L S; Rafferty, J A; Fairbairn, L J

2000-05-01

As a single gene defect in mature bone marrow cells, chronic granulomatous disease (X-CGD) represents a disorder which may be amenable to gene therapy by the transfer of the missing subunit into hemopoietic stem cells. In the majority of cases lack of Gp91-phox causes the disease. So far, studies involving transfer of Gp91-phox cDNA, including a phase I clinical trial, have yielded disappointing results. Most often, low titers of virus have been reported. In the present study we investigated the possible reasons for low titer amphotropic viral production. To investigate the effect of Gp91 cDNA on the efficiency of retroviral production from the packaging cell line, GP+envAm12, we constructed vectors containing either the native cDNA, truncated versions of the cDNA or a mutated form (LATG) in which the natural translational start codon was changed to a stop codon. Following derivation of clonal packaging cell lines, these were assessed for viral titer by RNA slot blot and analyzed by non-parametrical statistical analysis (Whitney-Mann U-test). An improvement in viral titer of just over two-fold was found in packaging cells containing the start-codon mutant of Gp91 and no evidence of truncated viral RNA was seen in these cells. Further analysis revealed the presence of rearranged forms of the provirus in Gp91-expressing cells, and the production of truncated, unpackaged viral RNA. Protein analysis revealed that LATG-transduced cells did not express full-length Gp91-phox, whereas those containing the wild-type cDNA did. However, a truncated protein was seen in ATG-transduced cells which was also present in wild type cells. No evidence for the presence of a negative transcriptional regulatory element was found from studies with the deletion mutants. A statistically significant effect of protein production on the production of virus from Gp91-expressing cells was found. Our data point to a need to restrict expression of the Gp91-phox protein and its derivatives in order to enhance retroviral production and suggest that improvements in current vectors for CGD gene therapy may need to include controlled, directed expression only in mature neutrophils.

Microstructure evolution of T91 irradiated in the BOR60 fast reactor

NASA Astrophysics Data System (ADS)

Jiao, Z.; Taller, S.; Field, K.; Yeli, G.; Moody, M. P.; Was, G. S.

2018-06-01

Microstructures of T91 neutron irradiated in the BOR60 reactor at five temperatures between 376 °C and 524 °C to doses between 15.4 and 35.1 dpa were characterized using transmission electron microscopy (TEM), scanning transmission electron microscopy (STEM), and atom probe tomography (APT). Type a<100> dislocation loops were observed at 376-415 °C and network dislocations dominated at 460 °C and 524 °C. Cavities appeared in a bimodal distribution with a high density of small bubbles less than 2 nm at irradiation temperatures between 376 °C and 415 °C. Small bubbles were also observed at 460 °C and 524 °C but cavities greater than 2 nm were absent. Enrichment of Cr, Ni, and Si at the grain boundary was observed at all irradiation temperatures. Radiation-induced segregation (RIS) of Cr, Ni and Si appeared to saturate at 17.1 dpa and 376 °C. The temperature dependence of RIS of Cr, Ni and Si at the grain boundary, which showed a peak Cr enrichment temperature of 460 °C and a lower peak Ni and Si enrichment temperature of ∼400 °C, was consistent with observations of RIS of Cr in proton irradiated T91, suggesting that the same RIS mechanism may also apply to BOR60 irradiated T91. G-phase and Cu-rich precipitates were observed at 376-415 °C but were absent at 460 °C and 524 °C. The absence of G-phase at 524 °C could be related to the minimal segregation of Ni and Si in that condition.

Microstructure evolution of T91 irradiated in the BOR60 fast reactor

DOE PAGES

Jiao, Z.; Taller, S.; Field, K.; ...

2018-03-14

In this paper, microstructures of T91 neutron irradiated in the BOR60 reactor at five temperatures between 376 °C and 524 °C to doses between 15.4 and 35.1 dpa were characterized using transmission electron microscopy (TEM), scanning transmission electron microscopy (STEM), and atom probe tomography (APT). Type a<100> dislocation loops were observed at 376–415 °C and network dislocations dominated at 460 °C and 524 °C. Cavities appeared in a bimodal distribution with a high density of small bubbles less than 2 nm at irradiation temperatures between 376 °C and 415 °C. Small bubbles were also observed at 460 °C and 524more » °C but cavities greater than 2 nm were absent. Enrichment of Cr, Ni, and Si at the grain boundary was observed at all irradiation temperatures. Radiation-induced segregation (RIS) of Cr, Ni and Si appeared to saturate at 17.1 dpa and 376 °C. The temperature dependence of RIS of Cr, Ni and Si at the grain boundary, which showed a peak Cr enrichment temperature of 460 °C and a lower peak Ni and Si enrichment temperature of ~400 °C, was consistent with observations of RIS of Cr in proton irradiated T91, suggesting that the same RIS mechanism may also apply to BOR60 irradiated T91. G-phase and Cu-rich precipitates were observed at 376–415 °C but were absent at 460 °C and 524 °C. Finally, the absence of G-phase at 524 °C could be related to the minimal segregation of Ni and Si in that condition.« less

Microstructure evolution of T91 irradiated in the BOR60 fast reactor

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jiao, Z.; Taller, S.; Field, K.

In this paper, microstructures of T91 neutron irradiated in the BOR60 reactor at five temperatures between 376 °C and 524 °C to doses between 15.4 and 35.1 dpa were characterized using transmission electron microscopy (TEM), scanning transmission electron microscopy (STEM), and atom probe tomography (APT). Type a<100> dislocation loops were observed at 376–415 °C and network dislocations dominated at 460 °C and 524 °C. Cavities appeared in a bimodal distribution with a high density of small bubbles less than 2 nm at irradiation temperatures between 376 °C and 415 °C. Small bubbles were also observed at 460 °C and 524more » °C but cavities greater than 2 nm were absent. Enrichment of Cr, Ni, and Si at the grain boundary was observed at all irradiation temperatures. Radiation-induced segregation (RIS) of Cr, Ni and Si appeared to saturate at 17.1 dpa and 376 °C. The temperature dependence of RIS of Cr, Ni and Si at the grain boundary, which showed a peak Cr enrichment temperature of 460 °C and a lower peak Ni and Si enrichment temperature of ~400 °C, was consistent with observations of RIS of Cr in proton irradiated T91, suggesting that the same RIS mechanism may also apply to BOR60 irradiated T91. G-phase and Cu-rich precipitates were observed at 376–415 °C but were absent at 460 °C and 524 °C. Finally, the absence of G-phase at 524 °C could be related to the minimal segregation of Ni and Si in that condition.« less

10 CFR 455.91 - Eligibility.

Code of Federal Regulations, 2010 CFR

2010-01-01

... part to fund, from non-Federal sources, energy conservation measures for eligible institutions; and (c... 10 Energy 3 2010-01-01 2010-01-01 false Eligibility. 455.91 Section 455.91 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION GRANT PROGRAMS FOR SCHOOLS AND HOSPITALS AND BUILDINGS OWNED BY UNITS OF LOCAL...

40 CFR 91.205 - Banking.

Code of Federal Regulations, 2010 CFR

2010-07-01

... three model years. Negative credits must be banked according to the requirements under § 91.207(c... negative credits under § 91.207 generates positive emission credits, when the following formula is used for... requirements of this part. Hydrocarbon Plus Oxides of Nitrogen Exhaust Emission Standards [Grams per kilowatt...

14 CFR 91.603 - Aural speed warning device.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 14 Aeronautics and Space 2 2012-01-01 2012-01-01 false Aural speed warning device. 91.603 Section... Operating Requirements for Large and Transport Category Aircraft § 91.603 Aural speed warning device. No... aural speed warning device that complies with § 25.1303(c)(1). ...

14 CFR 91.603 - Aural speed warning device.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 14 Aeronautics and Space 2 2010-01-01 2010-01-01 false Aural speed warning device. 91.603 Section... Operating Requirements for Large and Transport Category Aircraft § 91.603 Aural speed warning device. No... aural speed warning device that complies with § 25.1303(c)(1). ...

14 CFR 91.603 - Aural speed warning device.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 14 Aeronautics and Space 2 2014-01-01 2014-01-01 false Aural speed warning device. 91.603 Section... Operating Requirements for Large and Transport Category Aircraft § 91.603 Aural speed warning device. No... aural speed warning device that complies with § 25.1303(c)(1). ...

14 CFR 91.603 - Aural speed warning device.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 14 Aeronautics and Space 2 2011-01-01 2011-01-01 false Aural speed warning device. 91.603 Section... Operating Requirements for Large and Transport Category Aircraft § 91.603 Aural speed warning device. No... aural speed warning device that complies with § 25.1303(c)(1). ...

14 CFR 91.603 - Aural speed warning device.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 14 Aeronautics and Space 2 2013-01-01 2013-01-01 false Aural speed warning device. 91.603 Section... Operating Requirements for Large and Transport Category Aircraft § 91.603 Aural speed warning device. No... aural speed warning device that complies with § 25.1303(c)(1). ...

21 CFR 640.91 - Processing.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 21 Food and Drugs 7 2010-04-01 2010-04-01 false Processing. 640.91 Section 640.91 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS ADDITIONAL... intravenous injection. (c) Microbial contamination. All processing steps shall be conducted in a manner to...

Nuclear Data Sheets for A = 91

DOE Office of Scientific and Technical Information (OSTI.GOV)

Baglin, Coral M.

2013-10-15

Experimental nuclear structure and decay data for all known A=91 nuclides (As, Se, Br, Kr, Rb, Sr, Y, Zr, Nb, Mo, Tc, Ru, Rh, Pd) have been evaluated. This evaluation, covering data received by 1 September 2013, supersedes the 1998 evaluation by C. M. Baglin published in Nuclear Data Sheets86, 1 (1999) (15 December 1998 literature cutoff), and subsequent evaluations by C. M. Baglin added to the ENSDF database for Kr, Sr and Zr (29 December 2000 literature cutoff) and by B. Singh for {sup 91}Tc (6 November 2000 literature cutoff)

High Temperature Behavior of Cr3C2-NiCr Coatings in the Actual Coal-Fired Boiler Environment

NASA Astrophysics Data System (ADS)

Bhatia, Rakesh; Sidhu, Hazoor Singh; Sidhu, Buta Singh

2015-03-01

Erosion-corrosion is a serious problem observed in steam-powered electricity generation plants, and industrial waste incinerators. In the present study, four compositions of Cr3C2-(Ni-20Cr) alloy coating powder were deposited by high-velocity oxy-fuel spray technique on T-91 boiler tube steel. The cyclic studies were performed in a coal-fired boiler at 1123 K ± 10 K (850 °C ± 10 °C). X-ray diffraction, scanning electron microscopy/energy dispersive X-ray analysis and elemental mapping analysis techniques were used to analyze the corrosion products. All the coatings deposited on T-91 boiler tube steel imparted hot corrosion resistance. The 65 pctCr3C2 -35 pct (Ni-20Cr)-coated T-91 steel sample performed better than all other coated samples in the given environment.

Aquimonas voraii gen. nov., sp. nov., a novel gammaproteobacterium isolated from a warm spring of Assam, India.

PubMed

Saha, P; Krishnamurthi, S; Mayilraj, S; Prasad, G S; Bora, T C; Chakrabarti, T

2005-07-01

A bacterial strain designated GPTSA 20(T), which was isolated from a warm spring in Assam, India, was characterized by using a polyphasic approach. The cells were Gram-negative, aerobic rods, which could not utilize or produce acid from most of the carbohydrates tested. The predominant fatty acids were C(15:0) iso (25.04%), C(17:1) iso omega9c (19.28%), C(16:0) iso (17.73%) and C(11:0) iso 3-OH (9.34%). The G+C content was 75 mol%. From 16S rRNA gene sequence analysis (1433 nucleotides, continuous stretch), it was confirmed that strain GPTSA 20(T) belonged to the class 'Gammaproteobacteria'. The closest 16S rRNA gene sequence similarity found (98.2%) was with an uncultured bacterium clone, NB-03 (accession no. AB117707), from an autotrophic nitrifying biofilm. Among culturable bacteria, the closest sequence similarities were with Fulvimonas soli (93.0%), Silanimonas lenta (92.8%), Thermomonas hydrothermalis (92.4%), Frateuria aurantia (91.9%), Rhodanobacter lindaniclasticus (91.9%), Thermomonas haemolytica (91.9%) and Pseudoxanthomonas taiwanensis (91.8%); similarities of less than 91.8% were obtained with other members of the class 'Gammaproteobacteria'. From the biochemical, physiological, chemotaxonomic and phylogenetic analysis, it was clear that strain GPTSA 20(T) was quite different from members of known genera of the class 'Gammaproteobacteria'. Therefore, it is proposed that strain GPTSA 20(T) represents a novel species within a new genus, with the name Aquimonas voraii gen. nov., sp. nov. The type strain is GPTSA 20(T) (=MTCC 6713(T)=JCM 12896(T)).

Study on corrosion of metal materials in nitrate molten salts

NASA Astrophysics Data System (ADS)

Zhai, Wei; Yang, Bo; Li, Maodong; Li, Shiping; Xin, Mingliang; Zhang, Shuanghong; Huang, Guojia

2017-01-01

High temperature molten salts as a heat transfer heat storage medium has been more widely used in the field of concentrated solar thermal power generation. In the thermal heat storage system, metal material stability and performance at high temperatures are of one major limitation in increasing this operating temperature. In this paper, study on corrosion of 321H, 304, 316L, P91 metal materials in modified solar two molten salts. The corrosion kinetics of 304, 316L, 321H, P91 metal material in the modified solar two molten salts at 450°C, 500°C is also investigated. Under the same condition it was found that 304, 321H corroded at a rate of 40% less than P91. Spallation of corrosion products was observed on P91 steel, while no obvious observed on other kinds of stainless steel. Corrosion rates of 304, 321H, and 316L slowly increased with temperature. Oxidation mechanisms little varied with temperature. Corrosion products of metal materials observed at 450°C, 500°C were primarily Fe oxide and Fe, Cr oxide.

Multiplicity-dependent and nonbinomial efficiency corrections for particle number cumulants

NASA Astrophysics Data System (ADS)

Bzdak, Adam; Holzmann, Romain; Koch, Volker

2016-12-01

In this article we extend previous work on efficiency corrections for cumulant measurements [Bzdak and Koch, Phys. Rev. C 86, 044904 (2012), 10.1103/PhysRevC.86.044904; Phys. Rev. C 91, 027901 (2015), 10.1103/PhysRevC.91.027901]. We will discuss the limitations of the methods presented in these papers. Specifically we will consider multiplicity dependent efficiencies as well as nonbinomial efficiency distributions. We will discuss the most simple and straightforward methods to implement those corrections.

Multiplicity-dependent and nonbinomial efficiency corrections for particle number cumulants

DOE PAGES

Bzdak, Adam; Holzmann, Romain; Koch, Volker

2016-12-19

Here, we extend previous work on efficiency corrections for cumulant measurements [Bzdak and Koch, Phys. Rev. C 86, 044904 (2012)PRVCAN0556-281310.1103/PhysRevC.86.044904; Phys. Rev. C 91, 027901 (2015)PRVCAN0556-281310.1103/PhysRevC.91.027901]. We will then discuss the limitations of the methods presented in these papers. Specifically we will consider multiplicity dependent efficiencies as well as nonbinomial efficiency distributions. We will discuss the most simple and straightforward methods to implement those corrections.

Molecular breeding of transgenic rice plants expressing a bacterial chlorocatechol dioxygenase gene.

PubMed

Shimizu, Masami; Kimura, Tetsuya; Koyama, Takayoshi; Suzuki, Katsuhisa; Ogawa, Naoto; Miyashita, Kiyotaka; Sakka, Kazuo; Ohmiya, Kunio

2002-08-01

The cbnA gene encoding the chlorocatechol dioxygenase gene from Ralstonia eutropha NH9 was introduced into rice plants. The cbnA gene was expressed in transgenic rice plants under the control of a modified cauliflower mosaic virus 35S promoter. Western blot analysis using anti-CbnA protein indicated that the cbnA gene was expressed in leaf tissue, roots, culms, and seeds. Transgenic rice calluses expressing the cbnA gene converted 3-chlorocatechol to 2-chloromucote efficiently. Growth and morphology of the transgenic rice plants expressing the cbnA gene were not distinguished from those of control rice plants harboring only a Ti binary vector. It is thus possible to breed transgenic plants that degrade chloroaromatic compounds in soil and surface water.

Improved detergent-based recovery of polyhydroxyalkanoates (PHAs).

PubMed

Yang, Yung-Hun; Brigham, Christopher; Willis, Laura; Rha, ChoKyun; Sinskey, Anthony

2011-05-01

Extracting polyhydroxyalkanoate (PHA) polymer from bacterial cells often involves harsh conditions, including use of environmentally harmful solvents. We evaluated different detergents under various conditions to extract PHA from Ralstonia eutropha and Escherichia coli cells. Most detergents tested recovered highly pure PHA polymer from cells in amounts that depended on the percentage of polymer present in the cell. Detergents such as linear alkylbenzene sulfonic acid (LAS-99) produced a high yield of high purity polymer, and less detergent was needed compared to the amount of SDS to produce comparable yields. LAS-99 also has the advantage of being biodegradable and environmentally safe. Chemical extraction of PHA with detergents could potentially minimize or eliminate the need to use harsh organic solvents, thus making industrial PHA production a cleaner technology process. © Springer Science+Business Media B.V. 2011

gp-91 mediates histone deacetylase inhibition-induced cardioprotection

PubMed Central

Zhao, Ting C; Zhang, Ling X; Cheng, Guangmao; Liu, Jun T

2010-01-01

We have recently shown that the inhibition of histone deacetylases (HDAC) protects the heart against ischemia and reperfusion (I/R) injury. The mechanism by which HDAC inhibition induces cardioprotection remains unknown. We sought to investigate whether the genetic disruption of gp-91, a subunit of NADPH-oxidase, would mitigate cardioprotection of HDAC inhibition. Wild-type and gp-91−/− mice were treated with a potent inhibitor of HDACs, trichostatin A (TSA, 0.1mg/kg, i.p.). Twenty-four hours later, the perfused hearts were subjected to 30 min of ischemia and 30 min of reperfusion. HDAC inhibition in wild-type mice produced marked improvements in ventricular functional recovery and the reduction of infarct size. TSA-induced cardioprotection was eliminated with genetic deletion of gp91. Notably, Western blot and immunostaining displayed a significant increase in gp-91 in myocardium following HDAC inhibition, which resulted in a mildly subsequent increase in the production of reactive oxygen species (ROS). The pretreatment of H9c2 cardiomyoblasts with TSA (50 nmol/L) decreased cell necrosis and increased viability in response to simulated ischemia (SI), which was abrogated by the transfection of cells with gp-91 siRNA, but not by scrambled siRNA. Furthermore, treatment of PLB-985 gp91+/+cells with TSA increased the resistance to SI, which also diminished with genetic disruption of gp91 in gp91phox-deficient PLB-985 cells. TSA treatment inhibited the increased active caspase-3 in H9c2 cardiomyoblasts and PLB-985 gp91+/+cells exposed to SI, which were prevented by knockdown of gp-91 by siRNA. These results suggest that a cascade consisting of gp-91 and HDAC inhibition plays an essential role in orchestrating the cardioprotective effect. PMID:20433879

Differentially expressed microRNAs associated with changes of transcript levels in detoxification pathways and DDT-resistance in the Drosophila melanogaster strain 91-R.

PubMed

Seong, Keon Mook; Coates, Brad S; Kim, Do-Hyup; Hansen, Allison K; Pittendrigh, Barry R

2018-01-01

Dichloro-diphenyl-trichloroethane (DDT) resistance among arthropod species is a model for understanding the molecular adaptations in response to insecticide exposures. Previous studies reported that DDT resistance may involve one or multiple detoxification genes, such as cytochrome P450 monooxygenases (P450s), glutathione S-transferases (GSTs), esterases, and ATP binding cassette (ABC) transporters, or changes in the voltage-sensitive sodium channel. However, the possible involvement of microRNAs (miRNAs) in the post-transcriptional regulation of genes associated with DDT resistance in the Drosophila melanogaster strain 91-R remains poorly understood. In this study, the majority of the resulting miRNAs discovered in small RNA libraries from 91-R and the susceptible control strain, 91-C, ranged from 16-25 nt, and contained 163 precursors and 256 mature forms of previously-known miRNAs along with 17 putative novel miRNAs. Quantitative analyses predicted the differential expression of ten miRNAs between 91-R and 91-C, and, based on Gene Ontology and pathway analysis, these ten miRNAs putatively target transcripts encoding proteins involved in detoxification mechanisms. RT-qPCR validated an inverse correlation between levels of differentially-expressed miRNAs and their putatively targeted transcripts, which implies a role of these miRNAs in the differential regulation of detoxification pathways in 91-R compared to 91-C. This study provides evidence associating the differential expression of miRNAs in response to multigenerational DDT selection in Drosophila melanogaster and provides important clues for understanding the possible roles of miRNAs in mediating insecticide resistance traits.

Electrochemically Based Modules for Sterilization in the Field. Phase 2.

DTIC Science & Technology

1995-09-30

NO: DAMD 17-91-C-1105 TITLE: Electrochemically Based Modules for Sterilization in the Field PRINCIPAL INVESTIGATOR(S) : G. Duncan Hitchens, Ph.D. Tom...NUMBERS Electrochemically Based Modules for Sterilization in the Field 6. AUTHOR(S) DAMD17-91-C-1105 G. Duncan Hitchens, Ph.D. Tom C. Allen, Tom D. Rogers...of high concentration ozone as an alternative to ethylene oxide for use in medical sterilization systems. Evaluations of high concentration

10 CFR 501.62 - Petition contents.

Code of Federal Regulations, 2010 CFR

2010-01-01

... not available to OFE for review. (Department of Energy Organization Act, Pub. L. 95-91, 91 Stat. 565 (42 U.S.C. 7101 et seq.); Powerplant and Industrial Fuel Use Act of 1978, Pub. L. 95-620, 92 Stat. 3269 (42 U.S.C. 8301 et seq.); Omnibus Budget Reconciliation Act of 1981, Pub. L. 97-35; E.O. 12009, 42...

Inoculating Helianthus annuus (sunflower) grown in zinc and cadmium contaminated soils with plant growth promoting bacteria--effects on phytoremediation strategies.

PubMed

Marques, Ana P G C; Moreira, Helena; Franco, Albina R; Rangel, António O S S; Castro, Paula M L

2013-06-01

Plant growth promoting bacteria (PGPR) may help reducing the toxicity of heavy metals to plants in polluted environments. In this work the effects of inoculating metal resistant and plant growth promoting bacterial strains on the growth of Helianthus annuus grown in Zn and Cd spiked soils were assessed. The PGPR strains Ralstonia eutropha (B1) and Chrysiobacterium humi (B2) reduced losses of weight in metal exposed plants and induced changes in metal bioaccumulation and bioconcentration - with strain B2 decreasing up to 67% Zn accumulation and by 20% Zn bioconcentration factor (BCF) in the shoots, up to 64% Zn uptake and 38% Zn BCF in the roots, and up to 27% Cd uptake and 27% Cd BCF in plant roots. The impact of inoculation on the bacterial communities in the rhizosphere of the plant was also assessed. Bacterial community diversity decreased with increasing levels of metal contamination in the soil, but in rhizosphere soil of plants inoculated with the PGPR strains, a higher bacterial diversity was kept throughout the experimental period. Inoculation of sunflower, particularly with C. humi (B2), appears to be an effective way of enhancing the short term stabilization potential of the plant in metal contaminated land, lowering losses in plant biomass and decreasing aboveground tissue contamination. Copyright © 2013 Elsevier Ltd. All rights reserved.

Stability of Extemporaneously Compounded Dexamethasone in Glass and Plastic Bottles and Plastic Syringes

PubMed Central

Ensom, Mary H H; Décarie, Diane

2014-01-01

Background Dexamethasone is widely used to treat rheumatic and endocrine disorders and chemotherapy-induced nausea and vomiting. A palatable, alcohol-free liquid formulation, with a suitable concentration to allow reasonable administration volume, is available only via extemporaneous compounding. Objective: To evaluate the stability of dexamethasone suspensions in commercially available vehicles (Oral Mix and Oral Mix SF) in various types of containers after storage at 25°C and 4°C for up to 91 days. Methods: Dexamethasone suspensions (1 mg/mL) were prepared in Oral Mix and Oral Mix SF and then transferred to amber glass and plastic prescription bottles and plastic oral syringes. Suspensions in all 3 types of containers were stored at 25°C; suspensions in glass and plastic bottles were also stored at 4°C. Samples were collected weekly from each container up to 28 days and then every 2 weeks up to 91 days. The samples were analyzed by a validated, stability-indicating high-performance liquid chromatography − ultraviolet detection method. A suspension was considered stable if it maintained at least 90% of its initial dexamethasone concentration. Changes in colour, taste, odour, precipitation (and ease of resuspension), and pH were used to assess physical compatibility. Results: All suspensions maintained at least 96% of the original concentration for up to 91 days with storage at 25°C or at 4°C. No notable changes in colour, taste, odour, precipitation, or pH were observed over the 91-day period. Conclusion: Dexamethasone suspensions (1 mg/mL) in Oral Mix and Oral Mix SF, stored in amber glass or plastic bottles or plastic syringes at 25°C or in amber glass or plastic bottles at 4°C can be expected to remain stable for up to 91 days. PMID:25214658

A Methodology for Capturing and Analyzing Data from Technology Base Seminar Wargames

DTIC Science & Technology

1991-09-01

LONG RANGE FIRES LTC Angus USAFAS Dr. Brown ARO Mr. Campi CECOM LTC De Broux USACACDA COL James ARDEC Mr. Konick HDL Mr. Lavoie AFAU MAJ Martin ARDEC...85.17 2 SUPPLY DISTRIBUTION 81.92 3 RELIABILITY 91.89 1 TAC SUP TRANSPORT 77.91 5 TAC POL TRANSPORT 78.66 4 137 TABLE 7. QUESTION 7. ASSESS THE...C) Logistica nobility ) ( ) ( ) ( ) () ( ) ( Industrial Bass C) ( ) () () C) () ( Soldier Training hSO () ( ) () ( ) ( ) ) ( Nmnpouranagm nt

Molecular and Biochemical Characterization of the Natural Chromosome-Encoded Class A β-Lactamase from Pseudomonas luteola▿

PubMed Central

Doublet, Benoît; Robin, Frédéric; Casin, Isabelle; Fabre, Laëtitia; Le Fleche, Anne; Bonnet, Richard; Weill, François-Xavier

2010-01-01

Pseudomonas luteola (formerly classified as CDC group Ve-1 and named Chryseomonas luteola) is an unusual pathogen implicated in rare but serious infections in humans. A novel β-lactamase gene, blaLUT-1, was cloned from the whole-cell DNA of the P. luteola clinical isolate LAM, which had a weak narrow-spectrum β-lactam-resistant phenotype, and expressed in Escherichia coli. This gene encoded LUT-1, a 296-amino-acid Ambler class A β-lactamase with a pI of 6 and a theoretical molecular mass of 28.9 kDa. The catalytic efficiency of this enzyme was higher for cephalothin, cefuroxime, and cefotaxime than for penicillins. It was found to be 49% to 59% identical to other Ambler class A β-lactamases from Burkholderia sp. (PenA to PenL), Ralstonia eutropha (REUT), Citrobacter sedlakii (SED-1), Serratia fonticola (FONA and SFC-1), Klebsiella sp. (KPC and OXY), and CTX-M extended-spectrum β-lactamases. No gene homologous to the regulatory ampR genes of class A β-lactamases was found in the vicinity of the blaLUT-1 gene. The entire blaLUT-1 coding region was amplified by PCR and sequenced in five other genetically unrelated P. luteola strains (including the P. luteola type strain). A new variant of blaLUT-1 was found for each strain. These genes (named blaLUT-2 to blaLUT-6) had nucleotide sequences 98.1 to 99.5% identical to that of blaLUT-1 and differing from this gene by two to four nonsynonymous single nucleotide polymorphisms. The blaLUT gene was located on a 700- to 800-kb chromosomal I-CeuI fragment, the precise size of this fragment depending on the P. luteola strain. PMID:19884377

78 FR 34935 - Revisions to Operational Requirements for the Use of Enhanced Flight Vision Systems (EFVS) and to...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-11

... Transportation Safety Board OEM Original equipment manufacturer OpSpec Operation specification PAR Precision....1) B. Consolidate EFVS requirements in part 91 in a new section (Sec. 91.176) C. Establish equipment... single new section for organizational and regulatory clarity. Section 91.189 would be amended to permit...

Control of fire blight by Pseudomonas fluorescens A506 and Pantoea vagans C9-1 applied as single strains and mixed inocula

USDA-ARS?s Scientific Manuscript database

The biological control agents Pseudomonas fluorescens A506 and Pantoea vagans C9-1 were evaluated individually and in combination for the suppression of fire blight of pear or apple in ten field trials inoculated with the pathogen Erwinia amylovora. The formulation of pathogen inoculum applied to b...

Characterization of the Biosynthetic Operon for the Antibacterial Peptide Herbicolin in Pantoea vagans Biocontrol Strain C9-1 and Incidence in Pantoea Species

PubMed Central

Kamber, Tim; Lansdell, Theresa A.; Stockwell, Virginia O.; Ishimaru, Carol A.; Smits, Theo H. M.

2012-01-01

Pantoea vagans C9-1 is a biocontrol strain that produces at least two antibiotics inhibiting the growth of Erwinia amylovora, the causal agent of fire blight disease of pear and apple. One antibiotic, herbicolin I, was purified from culture filtrates of P. vagans C9-1 and determined to be 2-amino-3-(oxirane-2,3-dicarboxamido)-propanoyl-valine, also known as Nß-epoxysuccinamoyl-DAP-valine. A plasposon library was screened for mutants that had lost the ability to produce herbicolin I. It was shown that mutants had reduced biocontrol efficacy in immature pear assays. The biosynthetic gene cluster in P. vagans C9-1 was identified by sequencing the flanking regions of the plasposon insertion sites. The herbicolin I biosynthetic gene cluster consists of 10 coding sequences (CDS) and is located on the 166-kb plasmid pPag2. Sequence comparisons identified orthologous gene clusters in Pantoea agglomerans CU0119 and Serratia proteamaculans 568. A low incidence of detection of the biosynthetic cluster in a collection of 45 Pantoea spp. from biocontrol, environmental, and clinical origins showed that this is a rare trait among the tested strains. PMID:22504810

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tan, Lizhen; Kim, B. K.; Yang, Ying

Ferritic-martensitic steels such as T91 and NF616 are candidate materials for several nuclear applications. Here, this study evaluates radiation resistance of T91 and NF616 by examining their microstructural evolutions and hardening after the samples were irradiated in the Advanced Test Reactor to ~4.3 displacements per atom (dpa) at an as-run temperature of 469 °C. In general, this irradiation did not result in significant difference in the radiation-induced microstructures between the two steels. Compared to NF616, T91 had a higher number density of dislocation loops and a lower level of radiation-induced segregation, together with a slightly higher radiation-hardening. Unlike dislocation loopsmore » developed in both steels, radiation-induced cavities were only observed in T91 but remained small with sub-10 nm sizes. Lastly, other than the relatively stable M 23C 6, a new phase (likely Sigma phase) was observed in T91 and radiation-enhanced MX → Z phase transformation was identified in NF616. Laves phase was not observed in the samples.« less

Al-TiC in situ composite coating fabricated by low power pulsed laser cladding on AZ91D magnesium alloy

NASA Astrophysics Data System (ADS)

Yang, Liuqing; Li, Zhiyong; Zhang, Yingqiao; Wei, Shouzheng; Liu, Fuqiang

2018-03-01

Al + (Ti + B4C) composite coating was cladded on AZ91D magnesium alloy by a low power pulsed Nd-YAG laser. The Ti+B4C mixed powder is with the ratio of Ti: B4C = 5:1, which was then mixed with Al powder by weight fraction of 10%, 15% and 20%, respectively. Scanning electron microscopy, energy dispersive spectrometer and X-ray diffraction were used to study the microstructure, chemical composition and phase composition of the coating. Results showed that the coating had satisfied metallurgical bonding with the magnesium substrate. Al3Mg2, Al12Mg17, Al3Ti and TiC were formed by in-situ reaction. The coatings have micro-hardness of 348HV, which is about 5-6 times higher than that of AZ91D. The wear resistance and corrosion resistance of the coatings are enhanced with the addition of the mixed powder.

Interspecies Variation in the Functional Consequences of Mutation of Cytochrome c

PubMed Central

Josephs, Tracy M.; Hibbs, Moira E.; Ong, Lily; Morison, Ian M.; Ledgerwood, Elizabeth C.

2015-01-01

The naturally occurring human cytochrome c variant (G41S) is associated with a mild autosomal dominant thrombocytopenia (Thrombocytopenia Cargeeg) caused by dysregulation of platelet production. The molecular basis of the platelet production defect is unknown. Despite high conservation of cytochrome c between human and mouse (91.4% identity), introducing the G41S mutation into mouse cytochrome c in a knockin mouse (Cycs G41S/G41S) did not recapitulate the low platelet phenotype of Thrombocytopenia Cargeeg. While investigating the cause of this disparity we found a lack of conservation of the functional impact of cytochrome c mutations on caspase activation across species. Mutation of cytochrome c at residue 41 has distinct effects on the ability of cytochrome c to activate caspases depending on the species of both the cytochrome c and its binding partner Apaf-1. In contrast to our previous results showing the G41S mutation increases the ability of human cytochrome c to activate caspases, here we find this activity is decreased in mouse G41S cytochrome c. Additionally unlike wildtype human cytochrome c, G41S cytochrome c is unable to activate caspases in Xenopus embryo extracts. Taken together these results demonstrate a previously unreported species-specific component to the interaction of cytochrome c with Apaf-1. This suggests that the electrostatic interaction between cytochrome c and Apaf-1 is not the sole determinant of binding, with additional factors controlling binding specificity and affinity. These results have important implications for studies of the effects of cytochrome c mutations on the intrinsic apoptosis pathway. PMID:26086723

33 CFR Appendix A to Part 157 - Damage Assumptions, Hypothetical Outflows, and Cargo Tank Size and Arrangements

Code of Federal Regulations, 2014 CFR

2014-07-01

... in section 2 of this Appendix; W i for a segregated ballast tank may be taken equal to zero; C i...; C i for a segregated ballast tank may be taken equal to zero; EC15NO91.180 when b i is equal to or greater than t c, Ki is equal to zero; EC15NO91.181 when h i is equal to or greater than v s, Z i is equal...

JPRS Report, Near East & South Asia

DTIC Science & Technology

1991-05-01

sensitive positions in scientific Elections fields, which has enabled them to examine secrets per- 91AE0336A Amman JORDAN TIMES in English taining to...undertaking major Arab development projects, which establish the pillars of an Arab economic 91AE0338C Tel Aviv MA ’ARIV in Hebrew 8 Mar 91...stability of the Middle East. Toward this goal, it is 91AEO329D TelAviv ’ALHAMISHMAR in Hebrew reasonable that it have an interest in a pact with demo- 8

Does overwinter temperature affect maternal body composition and egg traits in yellow perch Perca flavescens?

PubMed

Feiner, Z S; Coulter, D P; Guffey, S C; Höök, T O

2016-04-01

Female yellow perch Perca flavescens exposed to three overwinter temperature regimes (4, 8 and 13° C) for 150 days spawned in markedly different proportions upon spring warming (37% of females in 4° C v. 64 and 91% in 8 and 13° C treatments, respectively), but exhibited no differences in fecundity, egg size or egg lipid content. Females held at 4° C also exhibited less within-clutch egg size variation than females held at 13° C. Moreover, eggs differed among temperature treatments in the overall proportions of 18 fatty acids, with the colder treatments resulting in potentially higher quality eggs containing more of the unsaturated fatty acids C16:1, C22:6-n3 and C18:2 cis. Female somatic condition also varied with temperature. Maternal somatic growth and protein content increased while lipid content decreased in 13° C compared to the colder treatments. There were, however, no differences among treatments in the fatty acid composition of maternal muscle. These results suggest that the temperatures experienced during winter may be less influential to P. flavescens egg size or number, which may exhibit relatively little plasticity in this species, but can alter both the number of females that spawn and the overall composition of eggs and maternal somatic tissues, which may have implications for future reproductive success. © 2016 The Fisheries Society of the British Isles.

Characterization of corrosion phenomena and kinetics on T91 ferritic/martensitic steel exposed at 450 and 550 °C to flowing Pb-Bi eutectic with 10-7 mass% dissolved oxygen

NASA Astrophysics Data System (ADS)

Tsisar, Valentyn; Schroer, Carsten; Wedemeyer, Olaf; Skrypnik, Aleksandr; Konys, Jürgen

2017-10-01

Corrosion behavior of two heats of T91 ferritic/martensitic steel, with slightly different Cr content, was investigated in flowing (2 m/s) Pb-Bi with 10-7 mass%O at 450 and 550 °C. The observed corrosion modes are: protective scale formation; accelerated oxidation and solution-based corrosion attack. Accelerated oxidation at 450 °C results in general metal recession of about 10 μm after ∼9000 h exposure and ∼15 μm at 550 °C after 2000 h. More severe and local solution-based corrosion results in a maximum depth of attack of 50-960 μm and 115-190 μm, correspondingly. Incubation period for solution-based corrosion is 500-5000 h at 450 °C and ≤500 h at 550 °C. The slightly higher chromium content in one of the heat of T91 steel prolongs the incubation period by improving the stability of the Cr-based oxide film.

Linkage disequilibrium between the CYP2C19*2,*17 and CYP2C9*1 alleles and impact of VKORC1, CYP2C9, CYP2C19 gene polymorphisms and gene-gene interactions on warfarin therapy.

PubMed

Khalighi, Koroush; Cheng, Gang; Mirabbasi, Seyedabbas; Khalighi, Bahar; Wu, Yin; Fan, Wuqiang

2017-01-01

Warfarin therapy is complicated by its large inter-individual and intra-individual variability. Both genetic and non-genetic factors can affect warfarin therapy. This study aims to investigate the allele distribution of VKORC1, CYP2C9 and CYP2C19, contribution of different allele variants and possible gene-gene interaction on warfarin therapy. Four hundreds and ninety-two patients were enrolled and single nucleotide polymorphisms for vitamin K epoxide reductase complex subunit 1 (VKORC1), cytochrome P450 CYP2C9 and cytochrome P450 CYP2C19 were genotyped. CYP2C9*1 allele is in complete linkage disequilibrium with CYP2C19*2 and CYP2C19*17 (D' = 1) in our study population. Patient with VKORC1-1639 G > A, CYP2C9*2 and CYP2C9*3 genetic variants need significant lower warfarin dose than patient with wild type allele of VKORC1 1639 G or CYP2C9*1. There is no significant differences between CYP2C19 allele variants for warfarin stable dose and INR > 5 event. Because of the complete linkage disequilibrium between CYP2C19*2,*17 and CYP2C9*1, patient with CYP2C19 *2/*2, *2/*17 and *17/*17 genotypes tend to have higher warfarin dose than patient with CYP2C19*1/*1 genotype. Stepwise regression analysis showed that VKORC1, CYP2C9, body mass index (BMI), age and gender were included as a factor significantly contributing to warfarin dose, whereas CYP2C19 did not contribute to warfarin dose. No statistically significant interaction between CYP2C9 and VKORC1 on warfarin dose and INR > 5 event was detected in univariate general linear model analysis. Our study suggests that polymorphic variants of VKORC1 and CYP2C9 affect warfarin dose independently, whereas CYP2C19 did not contribute to warfarin therapy.

Detection of Campylobacter jejuni in rectal swab samples from Rousettus amplexicaudatus in the Philippines.

PubMed

Hatta, Yuki; Omatsu, Tsutomu; Tsuchiaka, Shinobu; Katayama, Yukie; Taniguchi, Satoshi; Masangkay, Joseph S; Puentespina, Roberto; Eres, Eduardo; Cosico, Edison; Une, Yumi; Yoshikawa, Yasuhiro; Maeda, Ken; Kyuwa, Shigeru; Mizutani, Tetsuya

2016-09-01

Bats are the second diversity species of mammals and widely distributed in the world. They are thought to be reservoir and vectors of zoonotic pathogens. However, there is scarce report of the evidence of pathogenic bacteria kept in bats. The precise knowledge of the pathogenic bacteria in bat microbiota is important for zoonosis control. Thus, metagenomic analysis targeting the V3-V4 region of the 16S rRNA of the rectal microbiota in Rousettus amplexicaudatus was performed using high throughput sequencing. The results revealed that 103 genera of bacteria including Camplyobacter were detected. Campylobacter was second predominant genus, and Campylobacter coli and Campylobacter jejuni were identified in microbiome of R. amplexicaudatus. Campylobacteriosis is one of the serious bacterial diarrhea in human, and the most often implicated species as the causative agent of campylobacteriosis is C. jejuni. Therefore, we investigated the prevalence of C. jejuni in 91 wild bats with PCR. As a result of PCR assay targeted on 16S-23S intergenic spacer, partial genome of C. jejuni was detected only in five R. amplexicaudatus. This is the first report that C. jejuni was detected in bat rectal swab samples. C. jejuni is the most common cause of campylobacteriosis in humans, transmitted through water and contact with livestock animals. This result indicated that R. amplexicaudatus may be a carrier of C. jejuni.

40 CFR 91.513 - Administrative procedures for public hearing.

Code of Federal Regulations, 2010 CFR

2010-07-01

... such other means as the Administrator finds appropriate to provide notice to the public. (e) Filing and... pursuant to this section and § 91.512(c) must be filed with the Hearing Clerk of the Agency. Filing is... this section and § 91.512(b). If filing is to be accomplished by mailing, the documents must be sent to...

Selective sweep analysis in the genomes of the 91-R and 91-C Drosophila melanogaster strains reveals few of the ‘usual suspects’ in Dichlorodiphenyltrichloroethane (DDT) resistance

USDA-ARS?s Scientific Manuscript database

Adaptation of insect phenotypes for survival after exposure to xenobiotics can result from selection at multiple loci with additive genetic effects. A high level dichlorodiphenyltrichloroethane (DDT) resistance phenotype in the Drosophila melanogaster strain 91-R has resulted due to continuous labo...

77 FR 23475 - PJM Interconnection, L.L.C., Duke Energy Ohio, Inc., Duke Energy Kentucky, Inc; Notice of Filing

Federal Register 2010, 2011, 2012, 2013, 2014

2012-04-19

... DEPARTMENT OF ENERGY Federal Energy Regulatory Commission [Docket Nos. ER12-91-000, ER12-91-002, ER12-92-002] PJM Interconnection, L.L.C., Duke Energy Ohio, Inc., Duke Energy Kentucky, Inc; Notice of Filing Take notice that on April 5, 2012, Duke Energy Ohio, Inc. and Duke Energy Kentucky, Inc., tendered...

7 CFR 1728.201 - RUS Bulletin 1728H-701, RUS Specification for Wood Crossarms (Solid and Laminated), Transmission...

Code of Federal Regulations, 2010 CFR

2010-01-01

..., Standard Method for Analysis of Treated Wood and Treating Solutions by X-Ray Spectroscopy. (viii) A11-83...) C1-91, All Timber Products—Preservative Treatment by Pressure Processes. (x) C4-91, Poles... spaced. (x) Knots which have a maximum of 5/8 inch (1.59 cm) diameter may intersect pin holes in the...

7 CFR 1728.201 - RUS Bulletin 1728H-701, RUS Specification for Wood Crossarms (Solid and Laminated), Transmission...

Code of Federal Regulations, 2011 CFR

2011-01-01

..., Standard Method for Analysis of Treated Wood and Treating Solutions by X-Ray Spectroscopy. (viii) A11-83...) C1-91, All Timber Products—Preservative Treatment by Pressure Processes. (x) C4-91, Poles... spaced. (x) Knots which have a maximum of 5/8 inch (1.59 cm) diameter may intersect pin holes in the...

Preparation and corrosion resistance of electroless Ni-P/SiC functionally gradient coatings on AZ91D magnesium alloy

NASA Astrophysics Data System (ADS)

Wang, Hui-Long; Liu, Ling-Yun; Dou, Yong; Zhang, Wen-Zhu; Jiang, Wen-Feng

2013-12-01

In this paper, the protective electroless Ni-P/SiC gradient coatings on AZ91D magnesium alloy substrate were successfully prepared. The prepared Ni-P/SiC gradient coatings were characterized for its microstructure, morphology, microhardness and adhesion to the substrate. The deposition reaction kinetics was investigated and an empirical rate equation for electroless Ni-P/SiC plating on AZ91D magnesium alloy was developed. The anticorrosion properties of the Ni-P/SiC gradient coatings in 3.5 wt.% NaCl solution were evaluated by potentiodynamic polarization and electrochemical impedance spectroscopy (EIS) studies. The potentiodynamic polarization measurements revealed that the SiC concentration in the bath and heat treatment can influence the corrosion protection performance of electroless deposited Ni-P/SiC gradient coatings. EIS studies indicated that higher charge transfer resistance and slightly lower capacitance values were obtained for Ni-P/SiC gradient coatings compared to Ni-P coatings. The corrosion resistance of the Ni-P/SiC gradient coatings increases initially and decreases afterwards with the sustained increasing of immersion time in the aggressive medium. The electroless Ni-P/SiC gradient coatings can afford better corrosion protection for magnesium alloy substrate compared with Ni-P coatings.

Petrogenetic implications from ultramafic rocks and pyroxenites in ophiolitic occurrences of East Othris, Greece

NASA Astrophysics Data System (ADS)

Koutsovitis, P.; Magganas, A.

2012-04-01

Ultramafic rocks and pyroxenites in east Othris are included within ophiolitic units near the villages of Vrinena, Karavomilos, Pelasgia, Eretria, Agios Georgios, Aerino and Velestino. The first five ophiolitic occurrences are estimated to have been emplaced between the Oxfordian and Tithonian-Berriasian[1,2,3], while the latter two have been emplaced during the Eocene[4]. Ultramafic rocks include variably serpentinized harzburgites and lherzolites. Pyroxenites are usually found in the form of crosscutting veins within the harzburgites. Ultramafic rocks include depleted lherzolites, with Al2O3 ranging from 1.12 to 1.80 wt% and Cr from 3250 to 3290 ppm, as well as moderate to highly depleted serpentinized harzburgites, with Al2O3 ranging from 0.69 to 1.98 wt% and Cr from 2663 to 5582 ppm. Pyroxenites have generally higher Al2O3 ranging from 1.91 to 3.08 wt% and variable Cr ranging from 1798 to 3611 ppm. Lherzolites mostly include olivines (Fo=87.07-89.23) and clinopyroxenes (Mg#=85.71-90.12). Spinels from Eretria lherzolite (TiO2=0.02-0.08 wt%, Al2O3=36.06-42.45 wt%, Cr#=31.67-36.33) are compositionally similar with those of MORB peridotites[5], while those from Vrinena lherzolite (TiO2=0.16-0.43 wt%, Al2O3=6.90-22.12 wt%, Cr#=57.69-76.88) are similar to SSZ peridotites[5]. Serpentinized harzburgites include few olivines (Fo=90.51-91.15), enstatite porphyroclasts (Mg#=87.42-88.91), as well as fine grained enstatites of similar composition. Harzburgites from Pelasgia, Eretria and Agios Georgios include spinels (TiO2=0.03-0.08 wt%, Al2O3=23.21-31.58 wt%, Cr#=45.21-56.85) which do not clearly show if they are related with MORB or SSZ peridotites[5]. Spinels from Karavomilos harzburgite (TiO2=0.02-0.05 wt%, Al2O3=45.71-50.85 wt%, Cr#=16.84-22.32) are compositionally similar with MORB peridotites[5], whereas spinels from Vrinena harzburgite (TiO2=0.15-0.19 wt%, Al2O3=1.42-1.86 wt% Cr#=91.64-93.47) with SSZ peridotites[5]. Pyroxenites include clinopyroxenes (Mg#=84.25-91.78) but also enstatites (Mg#=88.37-91.47). Spinels have been analysed in pyroxenites from Aerino and Velestino (TiO2=0.79-1.07 wt%, Al2O3=10.88-18.46 wt% Cr#=60.74-70.78), indicating SSZ settings. Application of the olivine-spinel[6], olivine-augite[7], Cpx-Opx[8,9] geothermometers, yield equilibration temperatures of 961-1075 oC for lherzolites, 895-1084 oC for harzburgites and 990-1011 oC for pyroxenites. Our data indicate that the ophiolitic occurrences of Vrinena, Aerino and Velestino include ultramafic rocks and pyroxenites related to SSZ processes, while the other ophiolitic occurrences embrace ultramafic rocks which originated from a MORB-like setting, similar to west Othris ophiolites. It should be noted that even lherzolites have Cr and Y values similar to those of a highly depleted mantle source. A supra-subduction zone origin of the east Othris ophiolites, possibly with a slab rollback in the Pindos oceanic basin, may explain the different geotectonic environment affinities of the studied rocks.

Insights into the surface topology of polyhydroxyalkanoate synthase: self-assembly of functionalized inclusions.

PubMed

Hooks, David O; Rehm, Bernd H A

2015-10-01

The polyhydroxyalkanoate (PHA) synthase catalyzes the synthesis of PHA and remains attached to the hydrophobic PHA inclusions it creates. Although this feature is actively exploited to generate functionalized biobeads via protein engineering, little is known about the structure of the PHA synthase. Here, the surface topology of Ralstonia eutropha PHA synthase was probed to inform rational protein engineering toward the production of functionalized PHA beads. Surface-exposed residues were detected by conjugating biotin to inclusion-bound PHA synthase and identifying the biotin-conjugated lysine and cysteine residues using peptide fingerprinting analysis. The identified sites (K77, K90, K139, C382, C459, and K518) were investigated as insertion sites for the generation of new protein fusions. Insertions of FLAG epitopes into exposed sites K77, K90, K139, and K518 were tolerated, retaining >65 % of in vivo activity. Sites K90, K139, and K518 were also tested by insertion of the immunoglobulin G (IgG)-binding domain (ZZ), successfully producing PHA inclusions able to bind human IgG in vitro. Although simultaneous insertions of the ZZ domain into two sites was permissive, insertion at all three lysine sites inactivated the synthase. The K90/K139 double ZZ insertion had the optimum IgG-binding capacity of 16 mg IgG/g wet PHA beads and could selectively purify the IgG fraction from human serum. Overall, this study identified surface-exposed flexible regions of the PHA synthase which either tolerate protein/peptide insertions or are critical for protein function. This further elucidates the structure and function of PHA synthase and provides new opportunities for generating functionalized PHA biobeads.

Revisiting the Single Cell Protein Application of Cupriavidus necator H16 and Recovering Bioplastic Granules Simultaneously

PubMed Central

Kunasundari, Balakrishnan; Murugaiyah, Vikneswaran; Kaur, Gurjeet; Maurer, Frans H. J.; Sudesh, Kumar

2013-01-01

Cupriavidus necator H16 (formerly known as Hydrogenomonas eutropha) was famous as a potential single cell protein (SCP) in the 1970s. The drawback however was the undesirably efficient accumulation of non-nutritive polyhydroxybutyrate (PHB) storage compound in the cytoplasm of this bacterium. Eventually, competition from soy-based protein resulted in SCP not receiving much attention. Nevertheless, C. necator H16 remained in the limelight as a producer of PHB, which is a material that resembles commodity plastics such as polypropylene. PHB is a 100% biobased and biodegradable polyester. Although tremendous achievements have been attained in the past 3 decades in the efficient production of PHB, this bioplastic is still costly. One of the main problems has been the recovery of PHB from the cell cytoplasm. In this study, we showed for the first time that kilogram quantities of PHB can be easily recovered in the laboratory without the use of any solvents and chemicals, just by using the cells as SCP. In addition, the present study also demonstrated the safety and tolerability of animal model used, Sprague Dawley given lyophilized cells of C. necator H16. The test animals readily produced fecal pellets that were whitish in color, as would be expected of PHB granules. The pellets were determined to contain about 82-97 wt% PHB and possessed molecular mass of around 930 kg/mol. The PHB granules recovered biologically possessed similar molecular mass compared to chloroform extracted PHB [950 kg/mol]. This method now allows the production and purification of substantial quantities of PHB for various experimental trials. The method reported here is easy, does not require expensive instrumentation, scalable and does not involve extensive use of solvents and strong chemicals. PMID:24205250

Officer Career Development: Cross-Sectional Sample -- Fiscal Years 1986 - 1987

DTIC Science & Technology

1991-09-01

E9B7 - - - - - - 1 0545 4. Recruiting E9El1 E9Bl1 - - - - - - 1 0546 5. PG School E9E13 E9B13 - - - - - - 1 0547 NOTES: Respose sale iffes in SOResponse...been made to me. 7. I had accepted a job offer. 8. Net applicable-I knew that I would be self - employed. 9. Nct applicanle-I had not looked for a job...1 1635 8. Self -employ - - - - C9H C9H - - 1 16363 9. Not applicabl- - - - C91 C91 - - 1 1637 NOTES:3 F-118 PERSONNEL DISTRIBUTION AND CAREER

40 CFR 91.508 - Cumulative Sum (CumSum) procedure.

Code of Federal Regulations, 2010 CFR

2010-07-01

... family may be determined to be in noncompliance for purposes of § 91.510. H = The Action Limit. It is 5.0 × σ, and is a function of the standard deviation, σ. σ = is the sample standard deviation and is... Equation must be final deteriorated test results as defined in § 91.509(c). Ci = max[0 0R (Ci-1 + Xi − (FEL...

Formation of intermetallic compound coating on magnesium AZ91 cast alloy

NASA Astrophysics Data System (ADS)

Zhu, Tianping; Gao, Wei

2009-08-01

This study describes an intermetallic compound coating formed on AZ91 Mg cast alloy. The Al sputtered on AZ91 cast alloy reacted with substrate during a short period of heat treatment at 435°C, resulting in the formation of a continuous intermetallic compound layer. The short period treatment has the advantage of minimizing the negative effect on the microstructure of substrate and the mechanical properties, comparing with the reported diffusion coatings. DSC measurement and examination on the cross-section of Al sputtered samples show that local melting occurred along the Al/substrate interface at the temperature range between 430~435°C. The formation mechanism of intermetallic compound coating is proposed in terms of the local melting at Al/substrate interface. The salt water immersion test showed significant improvement in corrosion resistance of the intermetallic compound coated AZ91 cast alloy compared with the as-cast alloys.

A Novel Heme Pocket Hemoglobin Variant Associated with Normal Hematology: Hb Zara or α91(FG3)Leu→Ile (α2) (HBA2: c.274C > A).

PubMed

Trova, Sandro; Mereu, Paolo; Decandia, Luca; Cocco, Elena; Masala, Bruno; Manca, Laura; Pirastru, Monica

2015-08-13

We report a new hemoglobin (Hb) variant on the HBA2 gene, Hb Zara [α91(FG3)Leu→Ile (α2); HBA2: c.274C > A], which was found in a Caucasian man from Croatia. It was observed by routine cation exchange chromatography as an abnormal 21.8% fraction overlapping Hb A 2 , and associated with normal hematology. It was slightly unstable by the standard isopropanol precipitation test. DNA analysis revealed the CTT > ATT mutation at codon 91 on an α2 gene of a normal α-globin gene arrangement. This new variant represents the sixth described mutation at codon α91 and fourth on the α2 locus. As a result of the slight instability due to the significant role of the α91 residue in the α1β2 contact, the level of the Hb Zara variant was lower than levels observed for several stable variants codified by the α2 locus.

A Novel Heme Pocket Hemoglobin Variant Associated with Normal Hematology: Hb Zara or α91(FG3)Leu→Ile (α2) (HBA2: c.274C > A).

PubMed

Trova, Sandro; Mereu, Paolo; Decandia, Luca; Cocco, Elena; Masala, Bruno; Manca, Laura; Pirastru, Monica

2015-01-01

We report a new hemoglobin (Hb) variant on the HBA2 gene, Hb Zara [α91(FG3)Leu→Ile (α2); HBA2: c.274C > A], which was found in a Caucasian man from Croatia. It was observed by routine cation exchange chromatography as an abnormal 21.8% fraction overlapping Hb A2, and associated with normal hematology. It was slightly unstable by the standard isopropanol precipitation test. DNA analysis revealed the CTT > ATT mutation at codon 91 on an α2 gene of a normal α-globin gene arrangement. This new variant represents the sixth described mutation at codon α91 and fourth on the α2 locus. As a result of the slight instability due to the significant role of the α91 residue in the α1β2 contact, the level of the Hb Zara variant was lower than levels observed for several stable variants codified by the α2 locus.

Stability of pyrimethamine in a liquid dosage formulation stored for three months.

PubMed

Nahata, M C; Morosco, R S; Hipple, T F

1997-12-01

The stability of pyrimethamine in a liquid dosage formulation stored for up to three months was studies. Commercially available 25-mg pyrimethamine tablets were crushed with a mortar and pestle and mixed with a 1:1 mixture of Simple Syrup, NF, and 1% methylcellulose to yield a suspension with a pyrimethamine concentration of 2 mg/mL. The suspension was poured into 10 amber plastic and 10 amber glass prescription bottles; 5 plastic and 5 glass bottles were stored at 4 degrees C, and the remaining bottles were kept at 25 degrees C. Samples were collected at intervals up to 91 days and tested for pyrimethamine concentration by stability-indicating high-performance liquid chromatography. Pyrimethamine remained stable throughout the three-month study period under all conditions. At 4 degrees C, pyrimethamine concentrations remained above 96% of the initial concentration; at 25 degrees C, pyrimethamine concentrations remained above 91%. No substantial changes in pH were observed. Pyrimethamine was stable for at least 91 days in an oral suspension stored in plastic or glass prescription bottles at 4 or 25 degrees C.

Development of a microbial process for the recovery of petroleum oil from depleted reservoirs at 91-96°C.

PubMed

Arora, Preeti; Ranade, Dilip R; Dhakephalkar, Prashant K

2014-08-01

A consortium of bacteria growing at 91°C and above (optimally at 96°C) was developed for the recovery of crude oil from declining/depleted oil reservoirs having temperature of more than 91°C. PCR-DGGE-Sequencing analysis of 16S rRNA gene fragments of NJS-4 consortium revealed the presence of four strains identified as members of the genus Clostridium. The metabolites produced by NJS-4 consortium included volatile fatty acids, organic acids, surfactants, exopolysaccarides and CO2, which reduced viscosity, emulsified crude oil and increased the pressure that facilitated displacement of emulsified oil towards the surface. NJS-4 enhanced oil recovery by 26.7% and 10.1% in sand pack trials and core flood studies respectively in optimized nutrient medium comprised of sucrose and sodium acetate as carbon/energy source and urea as nitrogen source (pH 7-9, 96°C, and 4% salinity). Nutrient medium for MEOR was constituted using commercial grade cheap nutrients to improve the economic viability of MEOR process. Copyright © 2014 Elsevier Ltd. All rights reserved.

Loss of Parafollicular Cells during Gravitational Changes (Microgravity, Hypergravity) and the Secret Effect of Pleiotrophin

PubMed Central

Albi, Elisabetta; Curcio, Francesco; Spelat, Renza; Lazzarini, Andrea; Lazzarini, Remo; Cataldi, Samuela; Loreti, Elisabetta; Ferri, Ivana; Ambesi-Impiombato, Francesco Saverio

2012-01-01

It is generally known that bone loss is one of the most important complications for astronauts who are exposed to long-term microgravity in space. Changes in blood flow, systemic hormones, and locally produced factors were indicated as important elements contributing to the response of osteoblastic cells to loading, but research in this field still has many questions. Here, the possible biological involvement of thyroid C cells is being investigated. The paper is a comparison between a case of a wild type single mouse and a over-expressing pleiotrophin single mouse exposed to hypogravity conditions during the first animal experiment of long stay in International Space Station (91 days) and three similar mice exposed to hypergravity (2Gs) conditions. We provide evidence that both microgravity and hypergravity induce similar loss of C cells with reduction of calcitonin production. Pleiotrophin over-expression result in some protection against negative effects of gravity change. Potential implication of the gravity mechanic forces in the regulation of bone homeostasis via thyroid equilibrium is discussed. PMID:23284618

Elastic moduli of rock glasses under pressure to 8 kilobars and geophysical implications.

USGS Publications Warehouse

Meister, R.; Robertson, E.C.; Werke, R.W.; Raspet, R.

1980-01-01

Shear and longitudinal velocities were measured by the ultrasonic phase comparison method as a function of pressure to 8 kbar on synthetic glasses of basalt, andesite, rhyolite, and quartz composition and on natural obsidian. Velocities of most of the glasses decrease anomalously with pressure, but increasingly more-normal behavior occurs with decrease in SiO2 content. The pressure derivatives of rigidity and bulk modulus increase linearly, from -3.39 to -0.26 and from -5.91 to +2.09, respectively, with decrease in SiO2 content from 100 to 49%. The change from negative to positive in the pressure derivatives of both moduli and observed at Poisson's ratio of about 0.25 is consitent with the Smyth model for the anomalous elastic behavior of glass. If the temperature in the upper mantle is about 1500oC, tholeiitic basalt would be molten in accordance with the partial melt explanation for the low-velocity zone; at 1300oC and below, basalt would be in the glassy state, especially if more felsic than tholeiite. -Authors

Supplementation with an insoluble fiber obtained from carob pod (Ceratonia siliqua L.) rich in polyphenols prevents dyslipidemia in rabbits through SIRT1/PGC-1α pathway.

PubMed

Valero-Muñoz, María; Ballesteros, Sandra; Ruiz-Roso, Baltasar; Pérez-Olleros, Lourdes; Martín-Fernández, Beatriz; Lahera, Vicente; de Las Heras, Natalia

2017-12-22

To investigate the mechanism implicated in the effect of an insoluble fiber (obtained from carob pod) rich in polyphenols (IFCP) in lipid metabolism in the liver. Male New Zealand rabbits were fed with the following diets for 8 weeks: control diet (CT group), dyslipidemic diet supplemented with 0.5% cholesterol + 14% coconut oil (DL group) and dyslipidemic diet containing 0.5% cholesterol + 14% coconut oil plus 3% IFCP (DL + IFCP group). Dyslipidemic diet with IFCP was able to reduce development of mixed dyslipidemia, liver relative weight and collagen I protein expression compared to DL rabbits. Analyses of the main enzymes implicated in cholesterol and triglycerides metabolism revealed that IFCP increased hepatic concentration of 3-hydroxy-3-methylglutaryl-CoA reductase (HMG-CoA reductase) and cytochrome P450, family 7, subfamily a, polypeptide 1C (CYP7A1) (82.34, 114.42%, respectively) as well as protein expression of LDL receptor (42.48%) in DL rabbits. Importantly, IFCP also increased hepatic lipase (HL) levels (91.43%) and decreased glycerol phosphate acyltransferase (GPAT) and sterol regulatory element-binding protein 1C (SREBP1c) liver expression levels (20.38 and 41.20%, respectively). Finally, sirtuin 1 (SIRT1) and peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1α) hepatic expression increased in DL + IFCP group compared with DL (159.81 and 48.00%, respectively). These findings show that IFCP is able to abrogate the deleterious effects of hepatic dyslipidemia by modulating SIRT1 and PGC-1α pathways.

Propulsion Systems

DTIC Science & Technology

2011-03-31

Manual 91 - 710 Vol. 2 ( AFSPCMAN 91 - 710 V2). 8. Astrium EADS. 2010. cs.astrium.eads.net. April. 9. Astrium EADS. 2010. : http...of the High Energy Density Matter (HEDM) Conference. Albuquerque, NM, February 24-27, (Phillips Lab PL-CP- 91 -3003, p. 3 ). 104. Sullivan, D. J...Chapter 3 . DATES COVERED (From - To) 4. TITLE AND SUBTITLE 5a. CONTRACT NUMBER Chapter 18 - Propulsion Systems 5b. GRANT NUMBER 5c. PROGRAM ELEMENT

Human Services Students' Attitudes toward Working with an Inner-City Gang Member: The Case of Alphonse.

ERIC Educational Resources Information Center

Ancis, Julie R.; Burke, Mary C.

1997-01-01

Investigates the attitudes of 91 human service counseling students toward working with an inner-city gang member. Results reveal qualitative differences in students' self-perceived ability to work with this client. Presents implications for human service education and training. (MKA)

Employee Involvement and Organizational Citizenship: Implications for Labor Law Reform and "Lean Production."

ERIC Educational Resources Information Center

Cappelli, Peter; Rogovsky, Nikolai

1998-01-01

Organizational citizenship--behavior that promotes organizations without explicit reward--was measured in a survey of 512 employees and 91 supervisors. Involvement in work organization increased citizenship behavior indirectly by changing job characteristics. Involvement in decisions about employment practices had little or no effect. (SK)

Influence of mitochondrial efficiency on beef lean color stability

USDA-ARS?s Scientific Manuscript database

Loss of electrons in the electron transport chain has been implicated as a source of variation in feed efficiency of meat producing animals. The present study was conducted to evaluate the effects of electron loss during electron transport on beef lean color stability. Beef carcasses (n = 91) were...

G-protein-coupled receptor 91 and succinate are key contributors in neonatal postcerebral hypoxia-ischemia recovery.

PubMed

Hamel, David; Sanchez, Melanie; Duhamel, François; Roy, Olivier; Honoré, Jean-Claude; Noueihed, Baraa; Zhou, Tianwei; Nadeau-Vallée, Mathieu; Hou, Xin; Lavoie, Jean-Claude; Mitchell, Grant; Mamer, Orval A; Chemtob, Sylvain

2014-02-01

Prompt post-hypoxia-ischemia (HI) revascularization has been suggested to improve outcome in adults and newborn subjects. Other than hypoxia-inducible factor, sensors of metabolic demand remain largely unknown. During HI, anaerobic respiration is arrested resulting in accumulation of carbohydrate metabolic intermediates. As such succinate readily increases, exerting its biological effects via a specific receptor, G-protein-coupled receptor (GPR) 91. We postulate that succinate/GPR91 enhances post-HI vascularization and reduces infarct size in a model of newborn HI brain injury. The Rice-Vannucci model of neonatal HI was used. Succinate was measured by mass spectrometry, and microvascular density was evaluated by quantification of lectin-stained cryosection. Gene expression was evaluated by real-time polymerase chain reaction. Succinate levels rapidly increased in the penumbral region of brain infarcts. GPR91 was foremost localized not only in neurons but also in astrocytes. Microvascular density increased at 96 hours after injury in wild-type animals; it was diminished in GPR91-null mice leading to an increased infarct size. Stimulation with succinate led to an increase in growth factors implicated in angiogenesis only in wild-type mice. To explain the mode of action of succinate/GPR91, we investigated the role of prostaglandin E2-prostaglandin E receptor 4, previously proposed in neural angiogenesis. Succinate-induced vascular endothelial growth factor expression was abrogated by a cyclooxygenase inhibitor and a selective prostaglandin E receptor 4 antagonist. This antagonist also abolished succinate-induced neovascularization. We uncover a dominant metabolic sensor responsible for post-HI neurovascular adaptation, notably succinate/GPR91, acting via prostaglandin E2-prostaglandin E receptor 4 to govern expression of major angiogenic factors. We propose that pharmacological intervention targeting GPR91 could improve post-HI brain recovery.

Searching for Last Glacial Deep-Sea Polar Carbonates in the Ross Sea Continental slope and Their Relevance to Chronological Constraints

NASA Astrophysics Data System (ADS)

Brambati, A.; Bonaccorsi, R.; Quaia, T.; Busetti, M.

2003-12-01

Ice-proximal glacial marine sediments from the Antarctic continental margin retain ice rafting events as proxy record for change in the volume and extension of the Antarctic ice sheet throughout glacial-interglacial cycles. However, the sedimentary sequences from the Ross Sea continental margin remain relatively poorly understood and most research has been focused mainly on continental shelf sequences during the last past decades. We present a data set (i.e., X-ray lithology, Multi Sensor Core Logger physical data, and preservation of biogenic carbonates), obtained from six deep-sea cores (1991-1999 Italian Antarctic Research Programme, PNRA - Summer cruises). Specifically, the cores were collected from a) the central Eastern sector (i.e., Core ANTA95-89C, depth: 2056 m, length: 401 cm and Core ANTA99-c22, depth: 2650 m, length: 851 cm); b) the central Western sector (i.e., Core ANTA99-c23; water depth: 2158 m, length: 548 cm; and ANTA99-c24, water depth: 2750 m, length: 811 cm); and c) the North Western sector (i.e., Core ANTA91-08C, and ANTA91-02C) of the Ross Sea Continental slope. Well-preserved calcareous foraminifers (N. pachyderma, sx) in coarse-grained IRD materials sparsely occur and/or are concentrated in discrete layers (i.e., up to 22 cm-thick) of at least three cores (i.e., Cores ANTA91-08, ANTA91-02, and ANTA95-89C, e.g., at 217-238 cm-depth). Some carbonate layers were deposited during a period of time bracketing Stage3/Stage2. In Core 89C foraminifers are associated to multiple ice rafting episodes and likely occurred with oceanographic changes in the properties of slope water masses. The search of well-preserved, in situ-deposited, polar carbonates is demanded for a reliable C-14 AMS dating of late Pleistocene events in the Ross Sea.

Long-term strength and allowable stresses of grade 10Kh9MFB and X10CrMoVNb9-1 (T91/P91) chromium heat-resistant steels

NASA Astrophysics Data System (ADS)

Skorobogatykh, V. N.; Danyushevskiy, I. A.; Schenkova, I. A.; Prudnikov, D. A.

2015-04-01

Currently, grade X10CrMoVNb9-1 (T91, P91) and 10Kh9MFB (10Kh9MFB-Sh) chromium steels are widely applied in equipment manufacturing for thermal power plants in Russia and abroad. Compilation and comparison of tensile, impact, and long-term strength tests results accumulated for many years of investigations of foreign grade X10CrMoVNb9-1, T91, P91, and domestic grade 10Kh9MFB (10Kh9MFB-Sh) steels is carried out. The property identity of metals investigated is established. High strength and plastic properties of steels, from which pipes and other products are made, for operation under creep conditions are confirmed. Design characteristics of long-term strength on the basis of tests with more than one million of hour-samples are determined ( and at temperatures of 500-650°C). The table of recommended allowable stresses for grade 10Kh9MFB, 10Kh9MFB-SH, X10CrMoVNb9-1, T91, and P91 steels is developed. The long-time properties of pipe welded joints of grade 10Kh9MFB+10Kh9MFB, 10Kh9MFB-Sh+10Kh9MFB-Sh, X10CrMoVNb9-1+X10CrMoVNb9-1, P91+P91, T91+T91, 10Kh9MFB (10Kh9MFB-Sh)+X10CrMoVNb9-1(T/P91) steels is researched. The welded joint reduction factor is experimentally determined.

Behavior of ferritic/martensitic steels after n-irradiation at 200 and 300 °C

NASA Astrophysics Data System (ADS)

Matijasevic, M.; Lucon, E.; Almazouzi, A.

2008-06-01

High chromium ferritic/martensitic (F/M) steels are considered as the most promising structural materials for accelerator driven systems (ADS). One drawback that needs to be quantified is the significant hardening and embrittlement caused by neutron irradiation at low temperatures with production of spallation elements. In this paper irradiation effects on the mechanical properties of F/M steels have been studied and comparisons are provided between two ferritic/martensitic steels, namely T91 and EUROFER97. Both materials have been irradiated in the BR2 reactor of SCK-CEN/Mol at 300 °C up to doses ranging from 0.06 to 1.5 dpa. Tensile tests results obtained between -160 °C and 300 °C clearly show irradiation hardening (increase of yield and ultimate tensile strengths), as well as reduction of uniform and total elongation. Irradiation effects for EUROFER97 starting from 0.6 dpa are more pronounced compared to T91, showing a significant decrease in work hardening. The results are compared to our latest data that were obtained within a previous program (SPIRE), where T91 had also been irradiated in BR2 at 200 °C (up to 2.6 dpa), and tested between -170 °C and 300 °C. Irradiation effects at lower irradiation temperatures are more significant.

Water splitting-biosynthetic system with CO₂ reduction efficiencies exceeding photosynthesis.

PubMed

Liu, Chong; Colón, Brendan C; Ziesack, Marika; Silver, Pamela A; Nocera, Daniel G

2016-06-03

Artificial photosynthetic systems can store solar energy and chemically reduce CO2 We developed a hybrid water splitting-biosynthetic system based on a biocompatible Earth-abundant inorganic catalyst system to split water into molecular hydrogen and oxygen (H2 and O2) at low driving voltages. When grown in contact with these catalysts, Ralstonia eutropha consumed the produced H2 to synthesize biomass and fuels or chemical products from low CO2 concentration in the presence of O2 This scalable system has a CO2 reduction energy efficiency of ~50% when producing bacterial biomass and liquid fusel alcohols, scrubbing 180 grams of CO2 per kilowatt-hour of electricity. Coupling this hybrid device to existing photovoltaic systems would yield a CO2 reduction energy efficiency of ~10%, exceeding that of natural photosynthetic systems. Copyright © 2016, American Association for the Advancement of Science.

Evaluation of Corrective Action Team (CAT) Leader Training in Aeronautical Systems Division

DTIC Science & Technology

1991-09-01

00A DI EVALUATION OF CORRECTIVE ACTION TEAM ( CAT ) LEADER TRAINING IN AERONAUTICAL SYSTEMS DIVISION CA THESIS Kirk J. Streitrater, Captain, USAF AFIT...EVALUATION OF CORRECTIVE ACTION TEAM ( CAT ) LEADER TRAINING IN AERONAUTICAL SYSTEMS DIVISION THESIS Kirk J. Streitmater, Captain, USAF AFIT/GSM/LSR/91S-25...8217, , C- s :C AFIT/GSM/LSR/91S-25 EVALUATION OF CORRECTIVE ACTION TEAM ( CAT ) LEADER TRAINING IN AERONAUTICAL SYSTEMS DIVISION THESIS Presented to the

Mutational analyses of Aquifex pyrophilus DNA ligase define essential domains for self-adenylation and DNA binding activity.

PubMed

Lim, J H; Choi, J; Kim, W; Ahn, B Y; Han, Y S

2001-04-15

We constructed nine deletion mutants of NAD+-dependent DNA ligase from Aquifex pyrophilus to characterize the functional domains. All of DNA ligase deletion mutants were analyzed in biochemical assays for NAD+-dependent self-adenylation, DNA binding, and nick-closing activity. Although the mutant lsub1 (91-362) included the active site lysine (KxDG), self-adenylation was not shown. However, the mutants lsub6 (1-362), lsub7 (1-516), and lsub9 (1-635) showed the same adenylation activity as that of wild type. The lsub5 (91-719), which has the C-terminal domain (487-719) as to lsub4 (91-486), showed minimal adenylation activity. These results suggest that the presence of N-terminal 90 residues is essential for the formation of an enzyme-AMP complex, while C-terminal domain (487-719) appears to play a minimal role in adenylation. It was found that the presence of C-terminal domain (487-719) is indispensable for DNA binding activity of lsub5 (91-719). The mutant lsub9 (1-635) showed reduced DNA binding activity compared to that of wild type, suggesting the contribution of the domain (636-719) for the DNA binding activity. Thus, we concluded that the N-terminal 90 residues and C-terminal domain (487-719) of NAD+-dependent DNA ligase from A. pyrophilus are mutually indispensable for binding of DNA substrate.

28 CFR 91.54 - Applicability.

Code of Federal Regulations, 2010 CFR

2010-07-01

... major federal action that might significantly impact the quality of the human environment. The CEQ... activities that may significantly impact the environment. (c) Projects. Although VOI/TIS money cannot be used... Impact Review Procedures for VOI/TIS Grant Program Application to Voi/tis Grant Program § 91.54...

12 CFR 238.91 - Authority, purpose, and scope.

Code of Federal Regulations, 2013 CFR

2013-01-01

... SYSTEM (CONTINUED) SAVINGS AND LOAN HOLDING COMPANIES (REGULATION LL) Management Official Interlocks... situations where the management interlock likely would have an anticompetitive effect. (c) Scope. This... 12 Banks and Banking 4 2013-01-01 2013-01-01 false Authority, purpose, and scope. 238.91 Section...

24 CFR 91.315 - Strategic plan.

Code of Federal Regulations, 2010 CFR

2010-04-01

... CONSOLIDATED SUBMISSIONS FOR COMMUNITY PLANNING AND DEVELOPMENT PROGRAMS State Governments; Contents of Consolidated Plan § 91.315 Strategic plan. (a) General. For the categories described in paragraphs (b), (c), (d), (e), and (f) of this section, the consolidated plan must do the following: (1) Indicate the general...

Formation and mechanism of nanocrystalline AZ91 powders during HDDR processing

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, Yafen; Fan, Jianfeng, E-mail: fanjianfeng@tyu

2017-03-15

Grain sizes of AZ91 alloy powders were markedly refined to about 15 nm from 100 to 160 μm by an optimized hydrogenation-disproportionation-desorption-recombination (HDDR) process. The effect of temperature, hydrogen pressure and processing time on phase and microstructure evolution of AZ91 alloy powders during HDDR process was investigated systematically by X-ray diffraction, optical microscopy, scanning electron microscopy and transmission electron microscopy, respectively. The optimal HDDR process for preparing nanocrystalline Mg alloy powders is hydriding at temperature of 350 °C under 4 MPa hydrogen pressure for 12 h and dehydriding at 350 °C for 3 h in vacuum. A modified unreacted coremore » model was introduced to describe the mechanism of grain refinement of during HDDR process. - Highlights: • Grain size of the AZ91 alloy powders was significantly refined from 100 μm to 15 nm. • The optimal HDDR technology for nano Mg alloy powders is obtained. • A modified unreacted core model of grain refinement mechanism was proposed.« less

Development of biological platform for the autotrophic production of biofuels

NASA Astrophysics Data System (ADS)

Khan, Nymul

The research described herein is aimed at developing an advanced biofuel platform that has the potential to surpass the natural rate of solar energy capture and CO2 fixation. The underlying concept is to use the electricity from a renewable source, such as wind or solar, to capture CO 2 via a biological agent, such as a microbe, into liquid fuels that can be used for the transportation sector. In addition to being renewable, the higher rate of energy capture by photovoltaic cells than natural photosynthesis is expected to facilitate higher rate of liquid fuel production than traditional biofuel processes. The envisioned platform is part of ARPA-E's (Advanced Research Projects Agency - Energy) Electrofuels initiative which aims at supplementing the country's petroleum based fuel production with renewable liquid fuels that can integrate easily with the existing refining and distribution infrastructure (http://arpae. energy.gov/ProgramsProjects/Electrofuels.aspx). The Electrofuels initiative aimed to develop liquid biofuels that avoid the issues encountered in the current generation of biofuels: (1) the reliance of biomass-derived technologies on the inefficient process of photosynthesis, (2) the relatively energy- and resource-intensive nature of agronomic processes, and (3) the occupation of large areas of arable land for feedstock production. The process proceeds by the capture of solar energy into electrical energy via photovoltaic cells, using the generated electricity to split water into molecular hydrogen (H2) and oxygen (O2), and feeding these gases, along with carbon dioxide (CO2) emitted from point sources such as a biomass or coal-fired power plant, to a microbial bioprocessing platform. The proposed microbial bioprocessing platform leverages a chemolithoautotrophic microorganism (Rhodobacter capsulatus or Ralstonia eutropha) naturally able to utilize these gases as growth substrates, and genetically modified to produce a triterpene hydrocarbon fuel molecule (C30+ botryococcenes) native to the alga Botryococcus braunii . In addition to the genetic modification and bioreactor performance studies of these organisms for the production of botryococcene or squalene, the research examined the potential economic feasibility of the proposed platform through the use of bioreactor, microbial energetic models and experimentally measured growth yield and maintenance coefficients. In order to carry out an economic analysis, a process model was created in Aspen with the bioreactor at the center. This is presented in Chapter 2. The model looked at the effects of growth yield and maintenance coefficients of R. capsulatus and R. eutropha, reactor residence time, gas-liquid mass-transfer coefficients, gas composition and specific fuel productivity on the volumetric productivity and fuel yield on H2. It was found that the organism with the lowest maintenance coefficient performed better under very low growth rates evaluated in the model (based on residence time through the reactor) performed the best. The optimum parameter values were then used to determine the capital and operating costs for a 5000 bbl-fuel/day plant and the final fuel cost based on the Levelized Cost of Electricity (LCOE). It was found that under the assumptions used in this analysis and crude oil prices, the LCOE required for economic feasibility must be less than 2¢/kWh. While not feasible under current market prices and costs, this work identifies key variables impacting process cost and discusses potential alternative paths toward economic feasibility. This was the best case scenario of the two organisms evaluated, and an optimally suited organism with high growth yield and low maintenance coefficient should obviously improve the economics. This economic constraint will improve with the rise of fossil fuel prices, which should occur if the environmentally detrimental effects of their use are factored into the price, through higher taxation, for example. A review of the current status of metabolic engineering of chemolithoautotrophs is carried out in order to identify the challenges and likely routes to overcome them. This is presented in Chapter 3 of this dissertation. The initial metabolic engineering and bioreactor studies was carried out using a number of gene-constructs on R. capsulatus and R. eutropha. The gene-constructs consisted of Plac promoter followed by the triterpene synthase genes (SS or BS) and other upstream genes. A comparison of the production of triterpenes were done in the different growth modes that R. capsulatus was capable of growing---aerobic heterotrophic, anaerobic photoheterotrophic and aerobic chemoautotrophic. Autotrophic productivity could likely be improved much further by increasing the available mass-transfer of the reactor. These efforts are presented in Chapter 4 of this dissertation. (Abstract shortened by UMI.).

Author Correction: Implications of net energy-return-on-investment for a low-carbon energy transition

NASA Astrophysics Data System (ADS)

King, Lewis C.; van den Bergh, Jeroen C. J. M.

2018-04-01

In the version of this Analysis originally published, the value of the pessimistic EROI for the geothermal energy source in Table 1 was incorrectly given as 14:1; it should have read 9:1. This has now been corrected in all versions of the Analysis.

33 CFR Appendix A to Part 157 - Damage Assumptions, Hypothetical Outflows, and Cargo Tank Size and Arrangements

Code of Federal Regulations, 2012 CFR

2012-07-01

... section 2 of this Appendix; W i for a segregated ballast tank may be taken equal to zero; C i=Volume of a... segregated ballast tank may be taken equal to zero; EC15NO91.180 when b i is equal to or greater than t c, Ki is equal to zero; EC15NO91.181 when h i is equal to or greater than v s, Z i is equal to zero; b i...

33 CFR Appendix A to Part 157 - Damage Assumptions, Hypothetical Outflows, and Cargo Tank Size and Arrangements

Code of Federal Regulations, 2011 CFR

2011-07-01

... section 2 of this Appendix; W i for a segregated ballast tank may be taken equal to zero; C i=Volume of a... segregated ballast tank may be taken equal to zero; EC15NO91.180 when b i is equal to or greater than t c, Ki is equal to zero; EC15NO91.181 when h i is equal to or greater than v s, Z i is equal to zero; b i...

33 CFR Appendix A to Part 157 - Damage Assumptions, Hypothetical Outflows, and Cargo Tank Size and Arrangements

Code of Federal Regulations, 2010 CFR

2010-07-01

... section 2 of this Appendix; W i for a segregated ballast tank may be taken equal to zero; C i=Volume of a... segregated ballast tank may be taken equal to zero; EC15NO91.180 when b i is equal to or greater than t c, Ki is equal to zero; EC15NO91.181 when h i is equal to or greater than v s, Z i is equal to zero; b i...

40 CFR 763.91 - Operations and maintenance.

Code of Federal Regulations, 2014 CFR

2014-07-01

... CONTROL ACT ASBESTOS Asbestos-Containing Materials in Schools § 763.91 Operations and maintenance. (a... protection. Local education agencies must comply with either the OSHA Asbestos Construction Standard at 29 CFR 1926.1101, or the Asbestos Worker Protection Rule at 40 CFR 763.120, whichever is applicable. (c...

40 CFR 763.91 - Operations and maintenance.

Code of Federal Regulations, 2013 CFR

2013-07-01

... CONTROL ACT ASBESTOS Asbestos-Containing Materials in Schools § 763.91 Operations and maintenance. (a... protection. Local education agencies must comply with either the OSHA Asbestos Construction Standard at 29 CFR 1926.1101, or the Asbestos Worker Protection Rule at 40 CFR 763.120, whichever is applicable. (c...

40 CFR 763.91 - Operations and maintenance.

Code of Federal Regulations, 2011 CFR

2011-07-01

... CONTROL ACT ASBESTOS Asbestos-Containing Materials in Schools § 763.91 Operations and maintenance. (a... protection. Local education agencies must comply with either the OSHA Asbestos Construction Standard at 29 CFR 1926.1101, or the Asbestos Worker Protection Rule at 40 CFR 763.120, whichever is applicable. (c...

40 CFR 763.91 - Operations and maintenance.

Code of Federal Regulations, 2010 CFR

2010-07-01

... CONTROL ACT ASBESTOS Asbestos-Containing Materials in Schools § 763.91 Operations and maintenance. (a... protection. Local education agencies must comply with either the OSHA Asbestos Construction Standard at 29 CFR 1926.1101, or the Asbestos Worker Protection Rule at 40 CFR 763.120, whichever is applicable. (c...

40 CFR 763.91 - Operations and maintenance.

Code of Federal Regulations, 2012 CFR

2012-07-01

... CONTROL ACT ASBESTOS Asbestos-Containing Materials in Schools § 763.91 Operations and maintenance. (a... protection. Local education agencies must comply with either the OSHA Asbestos Construction Standard at 29 CFR 1926.1101, or the Asbestos Worker Protection Rule at 40 CFR 763.120, whichever is applicable. (c...

40 CFR 91.1006 - Manufacturer-owned exemption and precertification exemption.

Code of Federal Regulations, 2010 CFR

2010-07-01

... (CONTINUED) AIR PROGRAMS (CONTINUED) CONTROL OF EMISSIONS FROM MARINE SPARK-IGNITION ENGINES Exclusion and Exemption of Marine SI Engines § 91.1006 Manufacturer-owned exemption and precertification exemption. (a... “Emission Control Information;” (B) Full corporate name and trademark of manufacturer; (C) Engine...

21 CFR 640.91 - Processing.

Code of Federal Regulations, 2012 CFR

2012-04-01

... STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma Protein Fraction (Human) § 640.91 Processing. (a) Date... °C or colder. (e) Heat treatment. Heating of the final containers of Plasma Protein Fraction (Human... concentration of the product. (g) Incubation. All final containers of Plasma Protein Fraction (Human) shall be...

21 CFR 640.91 - Processing.

Code of Federal Regulations, 2011 CFR

2011-04-01

... STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma Protein Fraction (Human) § 640.91 Processing. (a) Date... °C or colder. (e) Heat treatment. Heating of the final containers of Plasma Protein Fraction (Human... concentration of the product. (g) Incubation. All final containers of Plasma Protein Fraction (Human) shall be...

21 CFR 640.91 - Processing.

Code of Federal Regulations, 2013 CFR

2013-04-01

... STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma Protein Fraction (Human) § 640.91 Processing. (a) Date... °C or colder. (e) Heat treatment. Heating of the final containers of Plasma Protein Fraction (Human... concentration of the product. (g) Incubation. All final containers of Plasma Protein Fraction (Human) shall be...

21 CFR 640.91 - Processing.

Code of Federal Regulations, 2014 CFR

2014-04-01

... STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma Protein Fraction (Human) § 640.91 Processing. (a) Date... °C or colder. (e) Heat treatment. Heating of the final containers of Plasma Protein Fraction (Human... concentration of the product. (g) Incubation. All final containers of Plasma Protein Fraction (Human) shall be...

The occurrence of fluor-wagnerite in UHT granulites and its implications towards understanding fluid regimes in the evolution of deep crust: a case study from the Eastern Ghats Belt, India

NASA Astrophysics Data System (ADS)

Das, Kaushik; Tomioka, Naotaka; Bose, Sankar; Ando, Jun-ichi; Ohnishi, Ichiro

2017-06-01

We report the occurrence of a rare phosphate mineral, fluor-wagnerite (Mg1.91-1.94Fe0.06-0.07Ca<0.01) (P0.99-1.00O4)(OH0.02-0.17F0.98-0.83) from the Eastern Ghats Belt of India, an orogenic belt evolved during Meso- to Neoproterozoic time. The host rock, i.e. high- to ultrahigh temperature (UHT) granulites ( 1000 °C, 8-9 kbar) of the studied area was retrogressed after emplacement to mid-crustal level (800-850 °C, 6-6.5 kbar) as deduced from their pressure -temperature histories. Based on mineral chemical data and micro-Raman analyses, we document an unusual high Mg-F-rich chemistry of the F-wagnerite, which occur both in peak metamorphic porphyroblastic assemblages as well as in the retrograde matrix assemblage. Therefore, in absence of other common phosphates like apatite, fluor-wagnerite can act as an indicator for the presence of F-bearing fluids for rocks with high X Mg and/or fO2. The occurrence of F-rich minerals as monitors for fluid compositions has important implications for the onset of biotite dehydration melting and hence melt production in the deep crust. We propose that fluor-wagnerite can occur as an accessory mineral associated with F-rich fluids in lower-mid crustal rocks, and F in coexisting minerals should be taken into consideration when reconciling the petrogenetic grid of biotite-dehydration melting.

Influence of the β-Mg17Al12 Phase Morphology on the Corrosion Properties Of Az91hp Magnesium Alloy

NASA Astrophysics Data System (ADS)

Guo, Lingling; Zhang, Jumei

2017-09-01

The morphology of β-Mg17Al12 phase and corrosion behavior of AZ91HP magnesium alloy after spheroidizing treatment were investigated by optical microcope electrochemical and immersion tests in 3.5% NaCl at 25°C. The results show that the coarse divorced eutectic phase of AZ91HP cast magnesium alloy dissolve into Mg matrix during the isothermal process at 415°C, and the lameller β phase precipitated from magnesium solid solution as perlite-type precipitation during the slowly cooling. Next, the spheroidizing treatment at different temperatures for 20h was carried out, and the lameller β phase were spheroridizing by dissolved themselves. After spheroidizing treatment at 300°C for 20h, many small granular β phase are scattering within the magnesium matrix. The corrosion properties of AZ91HP magnesium alloy in 3.5% NaCl decreased obviously after spheroidizing treatment, the polarization measurement of the alloy can be up to -1.412V from -1.56V of the cast. The β-Mg17Al12 phase act as a corrosion barrier and hinder corrosion propagation, if the second phase is in the form of a spherical morphology.

42 CFR 9.1 - Applicability and purpose.

Code of Federal Regulations, 2012 CFR

2012-10-01

... CARE FOR CHIMPANZEES HELD IN THE FEDERALLY SUPPORTED SANCTUARY SYSTEM § 9.1 Applicability and purpose. (a) General. The standards of care set forth in this part apply to the chimpanzee sanctuaries that... chimpanzee sanctuary system authorized by section 481C of the Public Health Service (PHS) Act, as amended (42...

42 CFR 9.1 - Applicability and purpose.

Code of Federal Regulations, 2013 CFR

2013-10-01

... CARE FOR CHIMPANZEES HELD IN THE FEDERALLY SUPPORTED SANCTUARY SYSTEM § 9.1 Applicability and purpose. (a) General. The standards of care set forth in this part apply to the chimpanzee sanctuaries that... chimpanzee sanctuary system authorized by section 481C of the Public Health Service (PHS) Act, as amended (42...

42 CFR 9.1 - Applicability and purpose.

Code of Federal Regulations, 2014 CFR

2014-10-01

... CARE FOR CHIMPANZEES HELD IN THE FEDERALLY SUPPORTED SANCTUARY SYSTEM § 9.1 Applicability and purpose. (a) General. The standards of care set forth in this part apply to the chimpanzee sanctuaries that... chimpanzee sanctuary system authorized by section 481C of the Public Health Service (PHS) Act, as amended (42...

10 CFR 2.4 - Definitions.

Code of Federal Regulations, 2011 CFR

2011-01-01

.... Department means the Department of Energy established by the Department of Energy Organization Act (Pub. L. 95-91, 91 Stat. 565 42 U.S.C. 7101 et seq.) to the extent that the Department, or its duly authorized..., officers and components and transferred to the U.S. Energy Research and Development Administration and to...

10 CFR 501.50 - Policy.

Code of Federal Regulations, 2010 CFR

2010-01-01

... (CONTINUED) ALTERNATE FUELS ADMINISTRATIVE PROCEDURES AND SANCTIONS Prohibition Rules and Orders § 501.50... operation consistent with reasonable fuel efficiency in an existing installation unless and until OFE adopts... Act, Pub. L. 95-91, 91 Stat. 565 (42 U.S.C. 7101 et seq.); Powerplant and Industrial Fuel Use Act of...

New Electronic Materials and CO2 Reduction.

DTIC Science & Technology

1988-02-02

REPORT DOCUMENTATION PAGE ! a ;t, C ’ SE -R .r N ’ D RE--R’tThVE "j f ’ .NUS UNCLASSIFIED APPROVED FOR PUBLIC RELEASE AD-A240 192 Lit R 11 II6 NOOO 14...by H12 have been carried out at 290C in a microreactor with a H2 /CO ration of 9/1. The catalysts studied were iron(III) oxide, iron(1I) diiron(1II...Methanation studies have been carried out at 290’C in a microreactor with a H 2 /CO ratio of 9/1. The catalysts studied were iron(III) oxide, iron(IT

Recombinant latcripin 11 of Lentinula edodes C91-3 suppresses the proliferation of various cancer cells.

PubMed

Gao, Yifan; Padhiar, Arshad Ahmed; Wang, Jia; Zhang, Wei; Zhong, Mintao; Liu, Ben; Kang, Zhijie; Wang, Xiaoli; Li, Xingyun; Huang, Min

2018-02-05

Lentinula edodes C91-3 is an edible mushroom that has demonstrated a remarkable anti-tumor effect in various cancer cells both in vitro and in vivo. In the present study, we report the ability of recombinant thioredoxin-like latcripin 11 (LP-11) of Lentinula edodes C91-3 to suppress the proliferation of various cancer cells. The LP-11 gene of Lentinula edodes C91-3 was cloned in the pET-32a(+) expression vector and expressed in a prokaryotic system. The expressed protein was refolded by gradual dialysis and purified by affinity gel filtration chromatography. The antioxidant activity of LP-11 was tested by 1,1-dipheny l-2-picrylhydrazyl (DPPH) assay. The anti-tumor activity of recombinant LP-11 was tested in eight kinds of tumor cell lines by CCK-8 assay. Recombinant LP-11 significantly suppressed the proliferation of various cancer cells, but not normal human umbilical vein endothelial cells. Human lymphoma U937 cells exhibited the most sensitivity to LP-11 protein. U937 cell apoptosis was assessed by Annexin V staining coupled with flow cytometry, and mitochondrial morphology was analyzed by light and electron microscopy. It was revealed that recombinant LP-11 induced apoptosis in human leukemic monocyte lymphoma U937 cells. Our findings suggest that recombinant LP-11 is a promising agent for the treatment of lymphoma. Copyright © 2017. Published by Elsevier B.V.

Resistivity Changes Due to Precipitation Effects in Fibre Reinforced Mg-Al-Zn-Mn Alloy

NASA Astrophysics Data System (ADS)

Kiehn, J.; Kainer, K. U.; Vostrý, P.; Stulíková, I.

1997-05-01

The change of electrical properties of alumina short fibre reinforced Mg-Al-Zn-Mn alloy AZ91D during isochronal annealing up to 300 °C is discussed. The Saffil® fibres were incorporated into the magnesium alloy by direct squeeze casting. The fibre distribution is random planar parallel to the flat faces of the dc four-point resistivity specimens machined from the solution treated castings. A sharp drop of resistivity between 140 and 260 °C is explained by the formation of incoherent -phase particles. Some practical recommendations concerning the use of alumina short fibre reinforced AZ91 alloy are made on the basis of the results obtained. Es werden die Änderungen der elektrischen Eigenschaften der aluminiumoxid-kurzfaserverstärkten Mg-Al-Zn-Mn Legierung AZ91D während isochroner Wärmebehandlungen bis 300 °C diskutiert. Das direkte Preßgießverfahren diente zur Herstellung der Saffil®-Faser Magnesium Verbundwerkstoffe. Die Proben zur Widerstandsmessung nach der Vier-Punkt Methode wurden durch spanende Bearbeitung aus den lösungsgeglühten Preßgußstücken herausgearbeitet, so daß sie regellose Faserverteilung in den Ebenen parallel zu den flachen Probenseiten aufwiesen. Ein starker Abfall des elektrischen Widerstands im Temperaturbereich zwischen 140 und 260 °C wird durch die Bildung inkohärenter β-Phase erklärt. Auf Grundlage der Ergebnisse werden einige Empfehlungen zur Anwendung der kurzfaserverstärkten Legierung AZ91 gegeben.

Drinking Locations Prior to Impaired Driving among College Students: Implications for Prevention

ERIC Educational Resources Information Center

Usdan, Stuart L.; Moore, Charity G.; Schumacher, Joseph E.; Talbott, Laura L.

2005-01-01

Drinking and driving is perhaps the most serious problem associated with heavy drinking among college students in the United States. In this study, the authors examined drinking locations prior to impaired driving in a college student sample. They administered the Impaired Driving Assessment to 91 college students identified as high risk for…

Effect of thermal exposure in helium on mechanical properties and microstructure of 316L and P91

NASA Astrophysics Data System (ADS)

Kunzova, Klara; Berka, Jan; Siegl, Jan; Hausild, Petr

2016-04-01

In this paper, the effects of high temperature exposure in air as well as in impure He on mechanical properties of 316L and P91 steels were investigated. The experimental programme was part of material design of new experimental facility - high temperature helium loop. Some of the specimens were exposed in air at 750 °C for up to 1000 h. Another set of specimens were exposed in impure helium containing 1 ppmv CO2, 2 ppmv O2, 35 ppmv CH4, 250 ppmv CO and 400 ppmv H2 at 750 °C for up to 1000 h. Metalographical analysis, tensile tests, fracture toughness and hardness tests of exposed and non-exposed specimens were carried out. After the exposure both in air and He, the ultimate tensile strength of P91 decreased significantly more than that of 316L. After the exposure in He, the fracture toughness of 316L was reduced to 60% while fracture toughness of P91 showed no significant changes. The hardness of P91 decreased with exposure time in air. The measurement of the hardness of 316L was very scattered the most probably due to the heterogeneities in microstructure, the trend was not possible to evaluate.

Effects of ambient temperature on egg and larval development of the invasive emerald ash borer (Coleoptera: Buprestidae): implications for laboratory rearing.

PubMed

Duan, Jian J; Watt, Tim; Taylor, Phil; Larson, Kristi; Lelito, Jonathan P

2013-10-01

The emerald ash borer, Agrilus planipennis Fairmaire, an invasive beetle from Asia causing large scale ash (Fraxinus) mortality in North America, has been extremely difficult to rear in the laboratory because of its long life cycle and cryptic nature of immature stages. This lack of effective laboratory-rearing methods has not only hindered research into its biology and ecology, but also mass production of natural enemies for biological control of this invasive pest. Using sticks from the alternate host plant, Fraxinus uhdei (Wenzig) Lingelsh, we characterized the stage-specific development time and growth rate of both emerald ash borer eggs and larvae at different constant temperatures (12-35 degrees C) for the purpose of developing effective laboratory-rearing methods. Results from our study showed that the median time for egg hatching decreased from 20 d at 20 degrees C to 7 d at 35 degrees C, while no emerald ash borer eggs hatched at 12 degrees C. The developmental time for 50% of emerald ash borer larvae advancing to third, fourth, and J-larval stages at 20 degrees C were 8.3, 9.1, and 12.3 wk, respectively, approximately two times longer than at 30 degrees C for the corresponding instars or stages. In contrast to 30 degrees C, however, the development times of emerald ash borer larvae advancing to later instars (from oviposition) were significantly increased at 35 degrees C, indicating adverse effects of this high temperature. The optimal range of ambient temperature to rear emerald ash borer larvae should be between 25-30 degrees C; however, faster rate of egg and larval development should be expected as temperature increases within this range.

A Cyclic Altered Peptide Analogue Based on Myelin Basic Protein 87-99 Provides Lasting Prophylactic and Therapeutic Protection Against Acute Experimental Autoimmune Encephalomyelitis.

PubMed

Emmanouil, Mary; Tseveleki, Vivian; Triantafyllakou, Iro; Nteli, Agathi; Tselios, Theodore; Probert, Lesley

2018-01-31

In this report, amide-linked cyclic peptide analogues of the 87-99 myelin basic protein (MBP) epitope, a candidate autoantigen in multiple sclerosis (MS), are tested for therapeutic efficacy in experimental autoimmune encephalomyelitis (EAE). Cyclic altered peptide analogues of MBP 87-99 with substitutions at positions 91 and/or 96 were tested for protective effects when administered using prophylactic or early therapeutic protocols in MBP 72-85 -induced EAE in Lewis rats. The Lys 91 and Pro 96 of MBP 87-99 are crucial T-cell receptor (TCR) anchors and participate in the formation of trimolecular complex between the TCR-antigen (peptide)-MHC (major histocompability complex) for the stimulation of encephalitogenic T cells that are necessary for EAE induction and are implicated in MS. The cyclic peptides were synthesized using Solid Phase Peptide Synthesis (SPPS) applied on the 9-fluorenylmethyloxycarboxyl/tert-butyl Fmoc/tBu methodology and combined with the 2-chlorotrityl chloride resin (CLTR-Cl). Cyclo(91-99)[Ala 96 ]MBP 87-99 , cyclo(87-99)[Ala 91,96 ]MBP 87-99 and cyclo(87-99)[Arg 91 , Ala 96 ]MBP 87-99 , but not wild-type linear MBP 87-99 , strongly inhibited MBP 72-85 -induced EAE in Lewis rats when administered using prophylactic and early therapeutic vaccination protocols. In particular, cyclo(87-99)[Arg 91 , Ala 96 ]MBP 87-99 was highly effective in preventing the onset and development of clinical symptoms and spinal cord pathology and providing lasting protection against EAE induction.

Complementation of a mutant cell line: central role of the 91 kDa polypeptide of ISGF3 in the interferon-alpha and -gamma signal transduction pathways.

PubMed Central

Müller, M; Laxton, C; Briscoe, J; Schindler, C; Improta, T; Darnell, J E; Stark, G R; Kerr, I M

1993-01-01

Mutants in complementation group U3, completely defective in the response of all genes tested to interferons (IFNs) alpha and gamma, do not express the 91 and 84 kDa polypeptide components of interferon-stimulated gene factor 3 (ISGF3), a transcription factor known to play a primary role in the IFN-alpha response pathway. The 91 and 84 kDa polypeptides are products of a single gene. They result from differential splicing and differ only in a 38 amino acid extension at the C-terminus of the 91 kDa polypeptide. Complementation of U3 mutants with cDNA constructs expressing the 91 kDa product at levels comparable to those observed in induced wild-type cells completely restored the response to both IFN-alpha and -gamma and the ability to form ISGF3. Complementation with the 84 kDa component similarly restored the ability to form ISGF3 and, albeit to a lower level, the IFN-alpha response of all genes tested so far. It failed, however, to restore the IFN-gamma response of any gene analysed. The precise nature of the DNA motifs and combination of factors required for the transcriptional response of all genes inducible by IFN-alpha and -gamma remains to be established. The results presented here, however, emphasize the apparent general requirement of the 91 kDa polypeptide in the primary transcriptional response to both types of IFN. Images PMID:7693454

Cyclic citrullinated MBP87-99 peptide stimulates T cell responses: Implications in triggering disease.

PubMed

Apostolopoulos, Vasso; Deraos, George; Matsoukas, Minos-Timotheos; Day, Stephanie; Stojanovska, Lily; Tselios, Theodore; Androutsou, Maria-Eleni; Matsoukas, John

2017-01-15

Amino acid mutations to agonist peptide epitopes of myelin proteins have been used to modulate immune responses and experimental autoimmune encephalomyelitis (EAE, animal model of multiple sclerosis). Such amino acid alteration are termed, altered peptide ligands (APL). We have shown that the agonist myelin basic protein (MBP) 87-99 epitope (MBP 87-99 ) with crucial T cell receptor (TCR) substitutions at positions 91 and 96 (K 91 ,P 96 (TCR contact residues) to R 91 ,A 96 ; [R 91 ,A 96 ]MBP 87-99 ) results in altered T cell responses and inhibits EAE symptoms. In this study, the role of citrullination of arginines in [R 91 ,A 96 ]MBP 87-99 peptide analog was determined using in vivo experiments in combination with computational studies. The immunogenicity of linear [Cit 91 ,A 96 ,Cit 97 ]MBP 87-99 and its cyclic analog - cyclo(87-99)[Cit 91 ,A 96 ,Cit 97 ]MBP 87-99 when conjugated to the carrier mannan (polysaccharide) were studied in SJL/J mice. It was found that mannosylated cyclo(87-99)[Cit 91 ,A 96 ,Cit 97 ]MBP 87-99 peptide induced strong T cell proliferative responses and IFN-gamma cytokine secretion compared with the linear one. Moreover, the interaction of linear and cyclic peptide analogs with the major histocompatibility complex (MHC II, H2-IA s ) and TCR was analyzed using molecular dynamics simulations at the receptor level, in order to gain a better understanding of the molecular recognition mechanisms that underly the different immunological profiles of citrullinated peptides compared to its agonist native counterpart MBP 87-99 epitope. The results demonstrate that the citrullination of arginine in combination with the backbone conformation of mutated linear and cyclic analogs are significant elements for the immune response triggering the induction of pro-inflammatory cytokines. Copyright © 2016 Elsevier Ltd. All rights reserved.

Utilizing Waste Thermocol Sheets and Rusted Iron Wires to Fabricate Carbon-Fe3O4 Nanocomposite Based Supercapacitors: Turning Wastes into Value-Added Materials.

PubMed

Vadiyar, Madagonda M; Liu, Xudong; Ye, Zhibin

2018-05-14

In the present work, we demonstrate the synthesis of porous activated carbon (specific surface area, 1,883 m2 g-1), Fe3O4 nanoparticles, and carbon-Fe3O4 nanocomposites using local waste thermocol sheets and rusted iron wires. The resulting carbon, Fe3O4 nanoparticles, and carbon-Fe3O4 composites are used as electrode materials for supercapacitor application. In particular, C-Fe3O4 composite electrodes exhibit a high specific capacitance of 1,375 F g-1 at 1 A g-1 and longer cyclic stability with 98 % of capacitance retention over 10,000 cycles. Subsequently, asymmetric supercapacitor, i. e., C-Fe3O4//Ni(OH)2/CNT device exhibits a high energy density of 91.1 Wh kg-1 and a remarkable cyclic stability, showing 98% of capacitance retention over 10,000 cycles. Thus, this work has important implications not only for the fabrication of low-cost electrodes for high-performance supercapacitors but also for the recycling of waste thermocol sheets and rust iron wires for value-added reuse. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Extremely high sulfate reduction, sediment oxygen demand and benthic nutrient flux associated with a large-scale artificial dyke and its implication to benthic-pelagic coupling in the Yeongsan River estuary, Yellow Sea.

PubMed

Kim, Sung-Han; Lee, Jae Seong; Hyun, Jung-Ho

2017-07-15

We investigated environmental impact of large-scale dyke on the sediment geochemistry, sulfate reduction rates (SRRs), sediment oxygen demand (SOD) and potential contribution of benthic nutrient flux (BNF) to primary production in the Yeongsan River estuary, Yellow Sea. The sediment near the dyke (YE1) with high organic carbon (C org ) content (>4%, dry wt.) was characterized by extremely high SOD (327mmolm -2 d -1 ) and SRRs (91-140mmolm -2 d -1 ). The sulfate reduction accounted for 73% of C org oxidation, and was responsible for strikingly high concentrations of NH 4 + (7.7mM), PO 4 3- (67μM) and HS - (487μM) in pore water. The BNF at YE1 accounted for approximately 200% of N and P required for primary production in the water column. The results present one of the most extreme cases that the construction of an artificial dyke may have profound impacts on the biogeochemical and ecological processes in coastal ecosystems. Copyright © 2017 Elsevier Ltd. All rights reserved.

Catalytic glycerol steam reforming for hydrogen production

NASA Astrophysics Data System (ADS)

Dan, Monica; Mihet, Maria; Lazar, Mihaela D.

2015-12-01

Hydrogen production from glycerol by steam reforming combine two major advantages: (i) using glycerol as raw material add value to this by product of bio-diesel production which is obtained in large quantities around the world and have a very limited utilization now, and (ii) by implication of water molecules in the reaction the efficiency of hydrogen generation is increased as each mol of glycerol produces 7 mol of H2. In this work we present the results obtained in the process of steam reforming of glycerol on Ni/Al2O3. The catalyst was prepared by wet impregnation method and characterized through different methods: N2 adsorption-desorption, XRD, TPR. The catalytic study was performed in a stainless steel tubular reactor at atmospheric pressure by varying the reaction conditions: steam/carbon ratio (1-9), gas flow (35 ml/min -133 ml/min), temperature (450-650°C). The gaseous fraction of the reaction products contain: H2, CH4, CO, CO2. The optimum reaction conditions as resulted from this study are: temperature 550°C, Gly:H2O ratio 9:1 and Ar flow 133 ml/min. In these conditions the glycerol conversion to gaseous products was 43% and the hydrogen yield was 30%.

28 CFR 91.10 - General.

Code of Federal Regulations, 2013 CFR

2013-07-01

... Initiative § 91.10 General. (a) Scope of boot camp program. Funding is appropriated in fiscal year 1995 to... Delinquency Prevention Act (42 U.S.C. 5601 et seq.) shall apply. (d) Eligibility. (1) Funding is available for both adult and juvenile boot camps. To be eligible for the funding of boot camps, states must comply...

40 CFR 91.315 - Analyzer initial calibration.

Code of Federal Regulations, 2013 CFR

2013-07-01

... in § 91.316(b). (c) Zero setting and calibration. Using purified synthetic air (or nitrogen), set the CO, CO2, NOX and HC analyzers at zero. Connect the appropriate calibrating gases to the analyzers and record the values. The same gas flow rates shall be used as when sampling exhaust. (d) Rechecking of zero...

40 CFR 91.315 - Analyzer initial calibration.

Code of Federal Regulations, 2014 CFR

2014-07-01

... in § 91.316(b). (c) Zero setting and calibration. Using purified synthetic air (or nitrogen), set the CO, CO2, NOX and HC analyzers at zero. Connect the appropriate calibrating gases to the analyzers and record the values. The same gas flow rates shall be used as when sampling exhaust. (d) Rechecking of zero...

40 CFR 91.315 - Analyzer initial calibration.

Code of Federal Regulations, 2012 CFR

2012-07-01

... in § 91.316(b). (c) Zero setting and calibration. Using purified synthetic air (or nitrogen), set the CO, CO2, NOX and HC analyzers at zero. Connect the appropriate calibrating gases to the analyzers and record the values. The same gas flow rates shall be used as when sampling exhaust. (d) Rechecking of zero...

40 CFR 91.315 - Analyzer initial calibration.

Code of Federal Regulations, 2010 CFR

2010-07-01

... in § 91.316(b). (c) Zero setting and calibration. Using purified synthetic air (or nitrogen), set the CO, CO2, NOX and HC analyzers at zero. Connect the appropriate calibrating gases to the analyzers and record the values. The same gas flow rates shall be used as when sampling exhaust. (d) Rechecking of zero...

40 CFR 91.315 - Analyzer initial calibration.

Code of Federal Regulations, 2011 CFR

2011-07-01

... in § 91.316(b). (c) Zero setting and calibration. Using purified synthetic air (or nitrogen), set the CO, CO2, NOX and HC analyzers at zero. Connect the appropriate calibrating gases to the analyzers and record the values. The same gas flow rates shall be used as when sampling exhaust. (d) Rechecking of zero...

10 CFR 501.60 - Purpose and scope.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 10 Energy 4 2010-01-01 2010-01-01 false Purpose and scope. 501.60 Section 501.60 Energy DEPARTMENT OF ENERGY (CONTINUED) ALTERNATE FUELS ADMINISTRATIVE PROCEDURES AND SANCTIONS Exemptions and.... 95-91, 91 Stat. 565 (42 U.S.C. 7101 et seq.); Powerplant and Industrial Fuel Use Act of 1978, Pub. L...

10 CFR 501.68 - Decision and order.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 10 Energy 4 2010-01-01 2010-01-01 false Decision and order. 501.68 Section 501.68 Energy DEPARTMENT OF ENERGY (CONTINUED) ALTERNATE FUELS ADMINISTRATIVE PROCEDURES AND SANCTIONS Exemptions and... Organization Act, Pub. L. 95-91, 91 Stat. 565 (42 U.S.C. 7101 et seq.); Powerplant and Industrial Fuel Use Act...

14 CFR 437.71 - Flight rules.

Code of Federal Regulations, 2013 CFR

2013-01-01

... rocket-powered flight, a permittee must confirm that all systems and operations necessary to ensure that... endanger the public. (c) A permittee may not operate a reusable suborbital rocket in a careless or reckless... operate a reusable suborbital rocket in areas designated in a Notice to Airmen under § 91.137, § 91.138...

14 CFR 437.71 - Flight rules.

Code of Federal Regulations, 2014 CFR

2014-01-01

... rocket-powered flight, a permittee must confirm that all systems and operations necessary to ensure that... endanger the public. (c) A permittee may not operate a reusable suborbital rocket in a careless or reckless... operate a reusable suborbital rocket in areas designated in a Notice to Airmen under § 91.137, § 91.138...

14 CFR 437.71 - Flight rules.

Code of Federal Regulations, 2011 CFR

2011-01-01

... rocket-powered flight, a permittee must confirm that all systems and operations necessary to ensure that... endanger the public. (c) A permittee may not operate a reusable suborbital rocket in a careless or reckless... operate a reusable suborbital rocket in areas designated in a Notice to Airmen under § 91.137, § 91.138...

14 CFR 437.71 - Flight rules.

Code of Federal Regulations, 2012 CFR

2012-01-01

... rocket-powered flight, a permittee must confirm that all systems and operations necessary to ensure that... endanger the public. (c) A permittee may not operate a reusable suborbital rocket in a careless or reckless... operate a reusable suborbital rocket in areas designated in a Notice to Airmen under § 91.137, § 91.138...

14 CFR 437.71 - Flight rules.

Code of Federal Regulations, 2010 CFR

2010-01-01

... rocket-powered flight, a permittee must confirm that all systems and operations necessary to ensure that... endanger the public. (c) A permittee may not operate a reusable suborbital rocket in a careless or reckless... operate a reusable suborbital rocket in areas designated in a Notice to Airmen under § 91.137, § 91.138...

Stability of amlodipine besylate in two liquid dosage forms.

PubMed

Nahata, M C; Morosco, R S; Hipple, T F

1999-01-01

To determine the stability of amlodipine besylate in two liquid dosage forms under refrigeration and at room temperature. Commercially available amlodipine tablets (Norvasc-Pfizer) were used to prepare two suspensions: one in extemporaneously prepared 1% methylcellulose in syrup (1:1), and another in equal volumes of commercially available OraPlus/OraSweet. Each suspension containing amlodipine 1 mg/mL was stored in 10 plastic prescription bottles; 5 were stored at 4 degrees C and 5 at 25 degrees C. Samples were collected immediately after preparation (day 0) and on days 7, 14, 28, 42, 56, 70, and 91. Amlodipine concentration was measured by stability-indicating HPLC method (n = 15). Research laboratory at Children's Hospital. Physical and chemical stability (> 90% of the initial concentration) of amlodipine in the two extemporaneously prepared suspensions during storage in plastic prescription bottles at 4 degrees C and 25 degrees C. Observed mean concentrations exceeded 90% of the initial concentrations in both suspensions for 91 days at 4 degrees C and 56 days at 25 degrees C. No noticeable change in physical appearance or odor was observed; pH changed slightly in the methylcellulose-containing formulation stored at 25 degrees C. Amlodipine was stable in two suspensions when stored in plastic prescription bottles for 91 days at 4 degrees C or 56 days at 25 degrees C. These formulations may be considered for pediatric or elderly patients who are unable to swallow tablets. The liquid dosage form would also permit accurate administration of amlodipine doses to infants and young children based on their body weight.

Diet and environment of a mid-Pliocene fauna in the Zanda Basin (western Himalaya): Paleo-elevation implications

NASA Astrophysics Data System (ADS)

Wang, Y.; Xu, Y.; Khawaja, S. N.; Wang, X.; Passey, B. H.; Zhang, C.; Li, Q.; Tseng, Z. J.; Takeuchi, G.; Deng, T.; Xie, G.

2011-12-01

A mid-Pliocene fauna (3.1-4.0 Ma) was recently discovered in the Zanda Basin in western Himalaya, at an elevation of about 4200 m above sea level. These fossil materials provide a unique window for examining the linkage among tectonic, climatic and biotic changes. Here we report the initial results from isotopic analyses of this fauna and of modern herbivores in the Zanda Basin. The δ13C values of enamel samples from modern wild Tibetan ass, horse, cow and goat from the Zanda Basin are -9.1±2.1%, which indicate a diet comprising predominantly of C3 plants and are consistent with the current dominance of C3 vegetation in the area. The enamel-δ13C values of the fossil horse, rhino, deer, and bovid are -9.6±0.8%, indicating that these ancient mammals, like modern herbivores in the area, fed primarily on C3 vegetation and lived in an environment dominated by C3 plants. The enamel-δ18O values of mid-Pliocene obligate drinkers (i.e., horse and rhino) are lower than those of their modern counterpart, most likely indicating a shift in climate to much drier conditions after ~3-4 Ma. Preliminary paleo-temperature estimates derived from a fossil-based temperature proxy as well as the "clumped isotope" thermometer for the mid-Pliocene Zanda Basin, although somewhat equivocal, are close to the present-day mean annual temperature in the area, suggesting that the paleo-elevation of the Zanda Basin in the mid-Pliocene was similar to its present-day elevation.

Final Environmental Assessment for Proposed Airspace Changes for Paradise East and Paradise West Military Operations Areas (MOAs) at Mountain Home Air Force Base (MHAFB) Idaho

DTIC Science & Technology

2010-03-29

IR FO R C E\\ 32 99 91 AI R SP A C E EA \\G IS \\M X D \\F IG U R E- A LT ER N AT IV E A. M X D M KO U BE K...basis. BO I \\\\O W L\\ PR O J\\ M O U N TA IN H O M EA IR FO R C E\\ 32 99 91 AI R SP A C E EA \\G IS \\M X D \\F IG U R E- A LT ER N AT IV E B. M X D M KO...MTR. MTR segments are identified and charted as either IR routes (IFR) or VR routes

Desulfovibrio oceani subsp. oceani sp. nov., subsp. nov. and Desulfovibrio oceani subsp. galateae subsp. nov., novel sulfate-reducing bacteria isolated from the oxygen minimum zone off the coast of Peru.

PubMed

Finster, Kai W; Kjeldsen, Kasper U

2010-03-01

Two deltaproteobacterial sulfate reducers, designated strain I.8.1(T) and I.9.1(T), were isolated from the oxygen minimum zone water column off the coast of Peru at 400 and 500 m water depth. The strains were Gram-negative, vibrio-shaped and motile. Both strains were psychrotolerant, grew optimally at 20 degrees C at pH 7.0-8.0 and at 2.5-3.5% NaCl (w/v). The strains grew by utilizing hydrogen/acetate, C(3-4) fatty acids, amino acids and glycerol as electron acceptors for sulfate reduction. Fumarate, lactate and pyruvate supported fermentative growth. Sulfate, sulfite, thiosulfate and taurin supported growth as electron acceptors. Both strains were catalase-positive and highly oxygen-tolerant, surviving 24 days of exposure to atmospheric concentrations. MK6 was the only respiratory quinone. The most prominent cellular fatty acid was iso-17:1-omega9c (18%) for strain I.8.1(T) and iso-17:0-omega9c (14%) for strain I.9.1(T). The G+C contents of their genomic DNA were 45-46 mol%. Phylogenetic analysis of 16S rRNA and dsrAB gene sequences showed that both strains belong to the genus Desulfovibrio. Desulfovibrio acrylicus DSM 10141(T) and Desulfovibrio marinisediminis JCM 14577(T) represented their closest validly described relatives with pairwise 16S rRNA gene sequence identities of 98-99%. The level of DNA-DNA hybridization between strains I.8.1(T) and I.9.1(T) was 30-38%. The two strains shared 10-26% DNA-DNA relatedness with D. acrylicus. Based on a polyphasic investigation it is proposed that strains I.8.1(T) and I.9.1(T) represent a novel species for which the name Desulfovibrio oceani sp. nov. is proposed with the two subspecies D. oceani subsp. oceani (type strain, I.8.1(T) = DSM 21390(T) = JCM 15970(T)) and D. oceani subsp. galateae (type strain, I.9.1(T) = DSM 21391(T) = JCM 15971(T)).

Lunatimonas lonarensis gen. nov., sp. nov., a haloalkaline bacterium of the family Cyclobacteriaceae with nitrate reducing activity.

PubMed

Srinivas, T N R; Aditya, S; Bhumika, V; Kumar, P Anil

2014-02-01

Novel pinkish-orange pigmented, Gram-negative staining, half-moon shaped, non-motile, strictly aerobic strains designated AK24(T) and AK26 were isolated from water and sediment samples of Lonar Lake, Buldhana district, Maharahstra, India. Both strains were positive for oxidase, catalase and β-galactosidase activities. The predominant fatty acids were iso-C15:0 (41.5%), anteiso-C15:0 (9.7%), iso-C17:0 3OH (9.6%), iso-C17:1 ω9c (10.2%) and C16:1 ω7c/C16:1 ω6c/iso-C15:0 2OH (summed feature 3) (14.4%). The strains contained MK-7 as the major respiratory quinone, and phosphatidylethanolamine and five unidentified lipids as the polar lipids. Blast analysis of the 16S rRNA gene sequence of strain AK24(T) showed that it was closely related to Aquiflexum balticum, with a pair-wise sequence similarity of 91.6%, as well as to Fontibacter ferrireducens, Belliella baltica and Indibacter alkaliphilus (91.3, 91.2 and 91.2% pair-wise sequence similarity, respectively), but it only had between 88.6 and 91.0% pair-wise sequence similarity to the rest of the family members. The MALDI-TOF assay reported no significant similarities for AK24(T) and AK26, since they potentially represented a new species. A MALDI MSP dendrogram showed close similarity between the two strains, but they maintained a distance from their phylogenetic neighbors. The genome of AK24(T) showed the presence of heavy metal tolerance genes, including the genes providing resistance to arsenic, cadmium, cobalt and zinc. A cluster of heat shock resistance genes was also found in the genome. Two lantibiotic producing genes, LanR and LasB, were also found in the genome of AK24(T). Strains AK24(T) and AK26 were very closely related to each other with 99.5% pair-wise sequence similarity. Phylogenetic analysis indicated that the strains were members of the family Cyclobacteriaceae and they clustered with the genus Mariniradius, as well as with the genera Aquiflexum, Cecembia, Fontibacter, Indibacter, and Shivajiella. DNA-DNA hybridization between strains AK24(T) and AK26 showed a relatedness of 82% and their rep-PCR banding patterns were very similar. Based on data from the current polyphasic study, it is proposed that the isolates be placed in a new genus and species with the name Lunatimonas lonarensis gen. nov., sp. nov. The type strain of Lunatimonas lonarensis is AK24(T) (=JCM 18822(T)=MTCC 11627(T)). Copyright © 2013 Elsevier GmbH. All rights reserved.

The Soviet Breakup and U.S. Foreign Policy.

ERIC Educational Resources Information Center

Lynch, Allen

1991-01-01

This issue of a quarterly publication on world affairs explores the historical significance of the disintegration of the Soviet Union and the implication for U.S. foreign policy. With the breakup of the USSR in 1990-91, Russia for the first time this century does not have control over the non-Russian nations of its former empire in Central Asia,…

Diagnostic value of MUC4 immunostaining in distinguishing epithelial mesothelioma and lung adenocarcinoma.

PubMed

Llinares, Karine; Escande, Fabienne; Aubert, Sébastien; Buisine, Marie-Pierre; de Bolos, Carme; Batra, Surinder K; Gosselin, Bernard; Aubert, Jean-Pierre; Porchet, Nicole; Copin, Marie-Christine

2004-02-01

The distinction between pleural malignant mesothelioma and pleural infiltration by adenocarcinomas has complex therapeutic and medicolegal implications. Although the panel of adenocarcinoma-associated antibodies and one or two mesothelioma markers is useful in this purpose, most of these antibodies are not totally specific. We determined the diagnostic value of MUC4 immunostaining in this issue. MUC4 gene expression was also studied by in situ hybridization and RT-PCR. MUC4 is a membrane-bound mucin that has been suggested to be implicated in malignant progression in humans and rats. The MUC4 gene is expressed in various normal epithelial tissues of endodermic origin and carcinomas. In the respiratory tract, MUC4 transcripts have been detected in normal respiratory epithelium and lung carcinomas. MUC4 protein was expressed in 32 of 35 (91.4%) lung adenocarcinomas on paraffin-embedded tissue. None of the 41 malignant mesotheliomas nor the 32 cases of benign mesothelial cells expressed MUC4 at the protein and mRNA levels. We conclude that MUC4 is a very specific (100%) and sensitive (91.4%) marker of lung adenocarcinomas on paraffin-embedded tissue that could be useful in diagnostic practice in the distinction between malignant mesothelioma and adenocarcinoma.

Proceedings of the 30th Annual Precise Time and Time Interval (PTTI) Systems and Applications Meeting

DTIC Science & Technology

1999-01-01

roa.es C. Tomis RedIRIS, Centro Comunicaciones CSIC RedIRIS Serrano 142, 28006 Madrid, Spain tel + 34-91-585-5150 fax +34-91-564-7421 e-mail...I&D) and managed by the Centro de Comunicaciones CSIC RedIRIS, which depends on the Scientific Research Council (Consejo Superior de Investigaciones

Prostate Dose Escalation by a Innovative Inverse Planning-Driven IMRT

DTIC Science & Technology

2008-11-01

density calibration was performed by scanning a phan- tom with inserts of known relative electron densities with respect to water (rwe ) and calibrating the...sim. 312, 91–112 2006. 11M. J. Murphy, “ Fracking moving organs in real time,” Semin. Radiat. Oncol. 141, 91–100 2004. 12P. C. Chi et al

14 CFR Appendix C to Part 25 - Appendix C to Part 25

Code of Federal Regulations, 2014 CFR

2014-01-01

... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Appendix C to Part 25 C Appendix C to Part... AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY AIRPLANES Pt. 25, App. C Appendix C to Part 25 Part I—Atmospheric....062 EC28SE91.063 (c) Takeoff maximum icing. The maximum intensity of atmospheric icing conditions for...

14 CFR Appendix C to Part 25 - Appendix C to Part 25

Code of Federal Regulations, 2012 CFR

2012-01-01

... 14 Aeronautics and Space 1 2012-01-01 2012-01-01 false Appendix C to Part 25 C Appendix C to Part... AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY AIRPLANES Pt. 25, App. C Appendix C to Part 25 Part I—Atmospheric....062 EC28SE91.063 (c) Takeoff maximum icing. The maximum intensity of atmospheric icing conditions for...

14 CFR Appendix C to Part 25 - Appendix C to Part 25

Code of Federal Regulations, 2010 CFR

2010-01-01

... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Appendix C to Part 25 C Appendix C to Part... AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY AIRPLANES Pt. 25, App. C Appendix C to Part 25 Part I—Atmospheric....062 EC28SE91.063 (c) Takeoff maximum icing. The maximum intensity of atmospheric icing conditions for...

Enhanced Incorporation of 3-Hydroxy-4-Methylvalerate Unit into Biosynthetic Polyhydroxyalkanoate Using Leucine as a Precursor

PubMed Central

2011-01-01

Ralstonia eutropha PHB-4 expressing Pseudomonas sp. 61-3 polyhydroxyalkanoate (PHA) synthase 1 (PhaC1Ps) synthesizes PHA copolymer containing 3-hydroxybutyrate (3HB) and a small amount (0.5 mol%) of 3-hydroxy-4-methylvalerate (3H4MV) from fructose as a carbon source. In this study, enhanced incorporation of 3H4MV into PHA was investigated using branched amino acid leucine as a precursor of 3H4MV. Leucine has the same carbon backbone as 3H4MV and is expected to be a natural and self-producible precursor. We found that the incorporation of 3H4MV was enhanced by the supplementation of excess amount (10 g/L) of leucine in the culture medium. This finding indicates that 3H4MV can be derived from leucine. To increase metabolic flux to leucine biosynthesis in the host strain by eliminating the feedback inhibition, the cells were subjected to N-methyl-N'-nitro-N-nitrosoguanidine (NTG) mutagenesis and leucine analog resistant mutants were generated. The mutants showed statistically higher 3H4MV fraction than the parent strain without supplementing leucine. Additionally, by supplying excess amount of leucine, the mutants synthesized 3HB-based PHA copolymer containing 3.1 mol% 3H4MV and 1.2 mol% 3-hydroxyvalerate (3HV) as minor constituents, which significantly affected the thermal properties of the copolymer. This study demonstrates that it is possible to enhance the monomer supply of 3H4MV into PHA by manipulating leucine metabolism. PMID:21906338

Characterization and evolution of an activator-independent methanol dehydrogenase from Cupriavidus necator N-1.

PubMed

Wu, Tung-Yun; Chen, Chang-Ting; Liu, Jessica Tse-Jin; Bogorad, Igor W; Damoiseaux, Robert; Liao, James C

2016-06-01

Methanol utilization by methylotrophic or non-methylotrophic organisms is the first step toward methanol bioconversion to higher carbon-chain chemicals. Methanol oxidation using NAD-dependent methanol dehydrogenase (Mdh) is of particular interest because it uses NAD(+) as the electron carrier. To our knowledge, only a limited number of NAD-dependent Mdhs have been reported. The most studied is the Bacillus methanolicus Mdh, which exhibits low enzyme specificity to methanol and is dependent on an endogenous activator protein (ACT). In this work, we characterized and engineered a group III NAD-dependent alcohol dehydrogenase (Mdh2) from Cupriavidus necator N-1 (previously designated as Ralstonia eutropha). This enzyme is the first NAD-dependent Mdh characterized from a Gram-negative, mesophilic, non-methylotrophic organism with a significant activity towards methanol. Interestingly, unlike previously reported Mdhs, Mdh2 does not require activation by known activators such as B. methanolicus ACT and Escherichia coli Nudix hydrolase NudF, or putative native C. necator activators in the Nudix family under mesophilic conditions. This enzyme exhibited higher or comparable activity and affinity toward methanol relative to the B. methanolicus Mdh with or without ACT in a wide range of temperatures. Furthermore, using directed molecular evolution, we engineered a variant (CT4-1) of Mdh2 that showed a 6-fold higher K cat/K m for methanol and 10-fold lower K cat/K m for n-butanol. Thus, CT4-1 represents an NAD-dependent Mdh with much improved catalytic efficiency and specificity toward methanol compared with the existing NAD-dependent Mdhs with or without ACT activation.

Long-term starvation and subsequent recovery of nitrifiers in aerated submerged fixed-bed biofilm reactors.

PubMed

Elawwad, Abdelsalam; Sandner, Hendrik; Kappelmeyer, Uwe; Koeser, Heinz

2013-01-01

The effectiveness of three operational strategies for maintaining nitrifiers in bench-scale, aerated, submerged fixed-bed biofilm reactors (SFBBRs) during long-term starvation at 20 degrees C were evaluated. The operational strategies were characterized by the resulting oxidation-reduction potential (ORP) in the SFBBRs. The activity rates of the nitrifiers were measured and the activity decay was expressed by half-life times. It was found that anoxic and alternating anoxic/aerobic conditions were the best ways to preserve ammonia-oxidizing bacteria (AOB) during long starvation periods and resulted in half-life times of up to 34 and 28 days, respectively. Extended anaerobic conditions caused the half-life for AOB to decrease to 21 days. In comparison, the activity decay of nitrite-oxidizing bacteria (NOB) tended to be slightly faster. The activity of AOB biofilms that were kept for 97 days under anoxic conditions could be completely recovered in less than one week, while over 4 weeks was needed for AOB kept under anaerobic conditions. NOB were more sensitive to starvation and required longer recovery periods than AOB. For complete recovery, NOB needed approximately 7 weeks, regardless of the starvation conditions applied. Using the fluorescence in situ hybridization (FISH) technique, Nitrospira was detected as the dominant NOB genus. Among the AOB, the terminal restriction fragment length polymorphism (TRFLP) technique showed that during starvation and recovery periods, the relative frequency of species shifted to Nitrosomonas europaea/eutropha, regardless of the starvation condition. The consequences of these findings for the operation of SFBBRs under low-load and starvation conditions are discussed.

Characterization of the Two CART Genes (CART1 and CART2) in Chickens (Gallus gallus)

PubMed Central

Huang, Long; Li, Juan; Wang, Yajun

2015-01-01

Cocaine- and amphetamine-regulated transcript (CART) peptide is implicated in the control of avian energy balance, however, the structure and expression of CART gene(s) remains largely unknown in birds. Here, we cloned and characterized two CART genes (named cCART1 and cCART2) in chickens. The cloned cCART1 is predicted to generate two bioactive peptides, cCART1(42-89) and cCART1(49-89), which share high amino acid sequence identity (94-98%) with their mammalian counterparts, while the novel cCART2 may produce a bioactive peptide cCART2(51-91) with 59% identity to cCART1. Interestingly, quantitative RT-PCR revealed that cCART1 is predominantly expressed in the anterior pituitary and less abundantly in the hypothalamus. In accordance with this finding, cCART1 peptide was easily detected in the anterior pituitary by Western blot, and its secretion from chick pituitaries incubated in vitro was enhanced by ionomycin and forskolin treatment, indicating that cCART1 is a novel peptide hormone produced by the anterior pituitary. Moreover, cCART1 mRNA expression in both the pituitary and hypothalamus is down-regulated by 48-h fasting, suggesting its expression is affected by energy status. Unlike cCART1, cCART2 is only weakly expressed in most tissues examined by RT-PCR, implying a less significant role of cCART2 in chickens. As in chickens, 2 or more CART genes, likely generated by gene and genome duplication event(s), were also identified in other non-mammalian vertebrate species including coelacanth. Collectively, the identification and characterization of CART genes in birds helps to uncover the roles of CART peptide(s) in vertebrates and provides clues to the evolutionary history of vertebrate CART genes. PMID:25992897

32 CFR Appendix C to Part 45 - DD Form 215

Code of Federal Regulations, 2010 CFR

2010-07-01

... 32 National Defense 1 2010-07-01 2010-07-01 false DD Form 215 C Appendix C to Part 45 National... CERTIFICATE OF RELEASE OR DISCHARGE FROM ACTIVE DUTY (DD FORM 214/5 SERIES) Pt. 45, App. C Appendix C to Part 45—DD Form 215 EC21OC91.050 ...

DOE Office of Scientific and Technical Information (OSTI.GOV)

Girardot, Crystal L.; Harlow, Donald G.

This document identifies 241-C Tank Farm (C Farm) leak causes and locations for the 100 series leaking tanks (241-C-101 and 241-C-105) identified in RPP-RPT-33418, Rev. 2, Hanford C-Farm Leak Inventory Assessments Report. This document satisfies the C Farm portion of the target (T04) in the Hanford Federal Facility Agreement and Consent Order milestone M-045-91F.

NOX2 amplifies acetaldehyde-mediated cardiomyocyte mitochondrial dysfunction in alcoholic cardiomyopathy

PubMed Central

Brandt, Moritz; Garlapati, Venkata; Oelze, Matthias; Sotiriou, Efthymios; Knorr, Maike; Kröller-Schön, Swenja; Kossmann, Sabine; Schönfelder, Tanja; Morawietz, Henning; Schulz, Eberhard; Schultheiss, Heinz-Peter; Daiber, Andreas; Münzel, Thomas; Wenzel, Philip

2016-01-01

Alcoholic cardiomyopathy (ACM) resulting from excess alcohol consumption is an important cause of heart failure (HF). Although it is assumed that the cardiotoxicity of the ethanol (EtOH)-metabolite acetaldehyde (ACA) is central for its development and progression, the exact mechanisms remain obscure. Murine cardiomyocytes (CMs) exposed to ACA or EtOH showed increased superoxide (O2•−) levels and decreased mitochondrial polarization, both being normalized by NADPH oxidase (NOX) inhibition. C57BL/6 mice and mice deficient for the ACA-degrading enzyme mitochondrial aldehyde dehydrogenase (ALDH-2−/−) were fed a 2% EtOH diet for 5 weeks creating an ACA-overload. 2% EtOH-fed ALDH-2−/− mice exhibited a decreased cardiac function, increased heart-to-body and lung-to-body weight ratios, increased cardiac levels of the lipid peroxidation product malondialdehyde (MDA) as well as increased NOX activity and NOX2/glycoprotein 91phox (NOX2/gp91phox) subunit expression compared to 2% EtOH-fed C57BL/6 mice. Echocardiography revealed that ALDH-2−/−/gp91phox−/− mice were protected from ACA-overload-induced HF after 5 weeks of 2% EtOH-diet, demonstrating that NOX2-derived O2•− contributes to the development of ACM. Translated to human pathophysiology, we found increased gp91phox expression in endomyocardial biopsies of ACM patients. In conclusion, ACM is promoted by ACA-driven mitochondrial dysfunction and can be improved by ablation of NOX2/gp91phox. NOX2/gp91phox therefore might be a potential pharmacological target to treat ACM. PMID:27624556

Detection and differentiation of Mycobacterium tuberculosis and nontuberculous mycobacterial isolates by real-time PCR.

PubMed

Shrestha, Nabin K; Tuohy, Marion J; Hall, Gerri S; Reischl, Udo; Gordon, Steven M; Procop, Gary W

2003-11-01

Mycobacteria cause a variety of illnesses that differ in severity and public health implications. The differentiation of Mycobacterium tuberculosis from nontuberculous mycobacteria (NTM) is of primary importance for infection control and choice of antimicrobial therapy. Despite advances in molecular diagnostics, the ability to rapidly diagnose M. tuberculosis infections by PCR is still inadequate, largely because of the possibility of false-negative reactions. We designed and validated a real-time PCR for mycobacteria by using the LightCycler system with 18 reference strains and 168 clinical mycobacterial isolates. All clinically significant mycobacteria were detected; the mean melting temperatures (with 99.9% confidence intervals [99.9% CI] in parentheses) for the different mycobacteria were as follows: M. tuberculosis, 64.35 degrees C (63.27 to 65.42 degrees C); M. kansasii, 59.20 degrees C (58.07 to 60.33 degrees C); M. avium, 57.82 degrees C (57.05 to 58.60 degrees C); M. intracellulare, 54.46 degrees C (53.69 to 55.23 degrees C); M. marinum, 58.91 degrees C (58.28 to 59.55 degrees C); rapidly growing mycobacteria, 53.09 degrees C (50.97 to 55.20 degrees C) or 43.19 degrees C (42.19 to 44.49 degrees C). This real-time PCR assay with melting curve analysis consistently accurately detected and differentiated M. tuberculosis from NTM. Detection of an NTM helps ensure that the negative result for M. tuberculosis is a true negative. The specific melting temperature also provides a suggestion of the identity of the NTM present, when the most commonly encountered mycobacterial species are considered. In a parallel comparison, both the LightCycler assay and the COBAS Amplicor M. tuberculosis assay correctly categorized 48 of 50 specimens that were proven by culture to contain M. tuberculosis, and the LightCycler assay correctly characterized 3 of 3 specimens that contained NTM.

Detection and Differentiation of Mycobacterium tuberculosis and Nontuberculous Mycobacterial Isolates by Real-Time PCR

PubMed Central

Shrestha, Nabin K.; Tuohy, Marion J.; Hall, Gerri S.; Reischl, Udo; Gordon, Steven M.; Procop, Gary W.

2003-01-01

Mycobacteria cause a variety of illnesses that differ in severity and public health implications. The differentiation of Mycobacterium tuberculosis from nontuberculous mycobacteria (NTM) is of primary importance for infection control and choice of antimicrobial therapy. Despite advances in molecular diagnostics, the ability to rapidly diagnose M. tuberculosis infections by PCR is still inadequate, largely because of the possibility of false-negative reactions. We designed and validated a real-time PCR for mycobacteria by using the LightCycler system with 18 reference strains and 168 clinical mycobacterial isolates. All clinically significant mycobacteria were detected; the mean melting temperatures (with 99.9% confidence intervals [99.9% CI] in parentheses) for the different mycobacteria were as follows: M. tuberculosis, 64.35°C (63.27 to 65.42°C); M. kansasii, 59.20°C (58.07 to 60.33°C); M. avium, 57.82°C (57.05 to 58.60°C); M. intracellulare, 54.46°C (53.69 to 55.23°C); M. marinum, 58.91°C (58.28 to 59.55°C); rapidly growing mycobacteria, 53.09°C (50.97 to 55.20°C) or 43.19°C (42.19 to 44.49°C). This real-time PCR assay with melting curve analysis consistently accurately detected and differentiated M. tuberculosis from NTM. Detection of an NTM helps ensure that the negative result for M. tuberculosis is a true negative. The specific melting temperature also provides a suggestion of the identity of the NTM present, when the most commonly encountered mycobacterial species are considered. In a parallel comparison, both the LightCycler assay and the COBAS Amplicor M. tuberculosis assay correctly categorized 48 of 50 specimens that were proven by culture to contain M. tuberculosis, and the LightCycler assay correctly characterized 3 of 3 specimens that contained NTM. PMID:14605148

Diagnostic utility of amyloid PET in cerebral amyloid angiopathy-related symptomatic intracerebral hemorrhage

PubMed Central

Baron, Jean-Claude; Farid, Karim; Dolan, Eamon; Turc, Guillaume; Marrapu, Siva T; O'Brien, Eoin; Aigbirhio, Franklin I; Fryer, Tim D; Menon, David K; Warburton, Elizabeth A; Hong, Young T

2014-01-01

By detecting β-amyloid (Aβ) in the wall of cortical arterioles, amyloid positron emission tomography (PET) imaging might help diagnose cerebral amyloid angiopathy (CAA) in patients with lobar intracerebral hemorrhage (l-ICH). No previous study has directly assessed the diagnostic value of 11C-Pittsburgh compound B (PiB) PET in probable CAA-related l-ICH against healthy controls (HCs). 11C-PiB-PET and magnetic resonance imaging (MRI) including T2* were obtained in 11 nondemented patients fulfilling the Boston criteria for probable CAA-related symptomatic l-ICH (sl-ICH) and 20 HCs without cognitive complaints or impairment. After optimal spatial normalization, cerebral spinal fluid (CSF)-corrected PiB distribution volume ratios (DVRs) were obtained. There was no significant difference in whole cortex or regional DVRs between CAA patients and age-matched HCs. The whole cortex DVR was above the 95% confidence limit in 4/9 HCs and 10/11 CAA patients (sensitivity=91%, specificity=55%). Region/frontal or occipital ratios did not have better discriminative value. Similar but less accurate results were found using visual analysis. In patients with sl-ICH, 11C-PiB-PET has low specificity for CAA due to the frequent occurrence of high 11C-PiB uptake in the healthy elderly reflecting incipient Alzheimer's disease (AD), which might also be present in suspected CAA. However, a negative PiB scan rules out CAA with excellent sensitivity, which has clinical implications for prognostication and selection of candidates for drug trials. PMID:24619277

Temperatures and locations used by hibernating bats, including Myotis sodalis (Indiana bat), in a limestone mine: implications for conservation and management.

PubMed

Brack, Virgil

2007-11-01

Understanding temperatures used by hibernating bats will aid conservation and management efforts for many species. A limestone mine with 71 km of passages, used as a hibernaculum by approximately 30,000 bats, was visited four times during a 6-year period. The mine had been surveyed and mapped; therefore, bats could be precisely located and temperatures (T (s)) of the entire hibernaculum ceiling accurately mapped. It was predicted that bats should hibernate between 5 and 10 degrees C to (1) use temperatures that allow a near minimal metabolic rate, (2) maximize the duration of hibernation bouts, (3) avoid more frequent and prolonged arousal at higher temperatures, (4) avoid cold and freezing temperatures that require an increase in metabolism and a decrease in duration of hibernation bouts or that could cause death, and (5) balance benefits of a reduced metabolic rate and costs of metabolic depression. The distribution of each species was not random for location (P < 0.000) or T (s) (P < 0.000). Myotis sodalis (Indiana bat) was most restricted in areas occupied, hibernating in thermally stable yet cold areas (X = 8.4 +/- 1.7 degrees C); 99% associated with cement block walls and sheltered alcoves, which perhaps dampened air movement and temperature fluctuations. Myotis lucifugus (little brown myotis) hibernated in colder, more variable areas (X = 7.2 +/- 2.6 degrees C). Myotis septentrionalis (northern myotis), Pipistrellus subflavus (eastern pipistrelle), and Eptesicus fuscus (big brown bat) typically hibernated in warm, thermally stable areas (X = 9.1 +/- 0.2 degrees C, X = 9.6 +/- 1.9 degrees C, and X = 9.5 +/- 1.5 degrees C, respectively). These data do not indicate that hibernacula for M. sodalis, an endangered species, should be manipulated to cool below 5 degrees C.

Implications of XRCC1, XPD and APE1 gene polymorphism in North Indian population: a comparative approach in different ethnic groups worldwide.

PubMed

Gangwar, Ruchika; Manchanda, Parmeet Kaur; Mittal, Rama Devi

2009-05-01

Identifying risk factors for human cancers should consider combinations of genetic variations and environmental exposures. Several polymorphisms in DNA repair genes have impact on repair and cancer susceptibility. We focused on X-ray repair cross-complementing group 1 (XRCC1), Xeroderma pigmentosum D (XPD) and apurinic/apyrimidinic endonuclease (APE1) as these are most extensively studied in cancer. Present study was conducted to determine distribution of XRCC1 C26304T, G27466A, G23591A, APE1 T2197G and XPD A35931C gene polymorphisms in North Indian population and compare with different populations globally. PCR-based analysis was conducted in 209 normal healthy individuals of similar ethnicity. Allelic frequencies in wild type of XRCC1 C26304T were 91.1% C(Arg); G27466A 62.9% G(Arg); G23591A 60.3% G(Arg); APE1 T2197G 75.1% T(Asp) and XPD A35931C 71.8% A(Lys). The variant allele frequency were 8.9% T(Trp) in XRCC1 C26304T; 37.1% A(His) in G27466A; 39.7% A(Gln) in G23591A; 24.9% G(Glu) in APE1 and 28.2% C(Gln) in XPD respectively. We further compared frequency distribution for these genes with various published studies in different ethnicity. Our results suggest that frequency in these DNA repair genes exhibit distinctive pattern in India that could be attributed to ethnicity variation. This could assist in high-risk screening of humans exposed to environmental carcinogens and cancer predisposition in different ethnic groups.

The Dental Fitness of Army War College Students, Class of 1985.

DTIC Science & Technology

1985-04-30

through April 1985) { TYPE OF TREATMENT RECEIVED # RECORDS N PT TX N OF APPTS EXAM PRO OPER C&B PROS PERIO OS ENDO 216 176 548 171 91 91 22 3 16 8 4 Notes...readiness. DEFINITIONS (3) DENTAL CONDITIONS are the oral or dental diseases, injuries , abnormalities, and situations which are regarded as pathologic...emerg 26 unknown 4 / E,O I Caries 62 unknown 4 / E,P,O I Caries 77 June 84 2 / E,O 2 Caries 82 July 77(84) 4 / EP,O I Caries 84 unknown 13 / E,O,C&B, Endo

Characterization of a polyhydroxyalkanoate obtained from pineapple peel waste using Ralsthonia eutropha.

PubMed

Vega-Castro, Oscar; Contreras-Calderon, Jose; León, Emilson; Segura, Almir; Arias, Mario; Pérez, León; Sobral, Paulo J A

2016-08-10

Agro-industrial waste can be the production source of biopolymers such as polyhydroxyalkanoates. The aim of this study was to produce and characterize Polyhydroxyalkanoates produced from pineapple peel waste fermentation processes. The methodology includes different pineapple peel waste fermentation conditions. The produced biopolymer was characterized using FTIR, GC-MS and NMR. The best fermentation condition for biopolymer production was obtained using pH 9, Carbon/Nitrogen 11, carbon/phosphorus 6 and fermentation time of 60h. FTIR analyzes showed PHB group characteristics, such as OH, CH and CO. In addition, GC-MS showed two monomers with 4 and 8 carbons, referred to PHB and PHBHV. H(1) NMR analysis showed 0.88-0.97 and 5.27ppm signals, corresponding to CH3 and CH, respectively. In conclusion, polyhydroxyalkanoate production from pineapple peels waste is an alternative for the treatment of waste generated in Colombia's fruit industry. Copyright © 2016 Elsevier B.V. All rights reserved.

Characterization of bacterial structures in two-stage moving-bed biofilm reactor (MBBR) during nitrification of the landfill leachate.

PubMed

Ciesielski, Slawomir; Kulikowska, Dorota; Kaczowka, Ewelina; Kowal, Przemysław

2010-07-01

Differences in DNA banding patterns, obtained by ribosomal intergenic spacer analysis (RISA), and nitrification were followed in a moving-bed biofilm reactor (MBBR) receiving municipal landfill leachate. Complete nitrification (> 99%) to nitrate was obtained in the two-stage MBBR system with an ammonium load of 1.09 g N-NH(4)/m(2).d. Increasing the ammonium load to 2.03 g N-NH(4)/m(2).d or more caused a decline in process efficiency to 70-86%. Moreover, at the highest ammonium load (3.76 g N-NH(4)/m(2).d), nitrite was the predominant product of nitrification. Community succession was evident in both compartments in response to changes in ammonium load. Non-metric multidimensional scaling (NMDS) supported by similarity analysis (ANOSIM) showed that microbial biofilm communities differed between compartments. The microbial biofilm was composed mainly of ammonia-oxidizing bacteria (AOB), with Nitrosomonas europeae and N. eutropha being most abundant. These results suggest that high ammonium concentrations select for particular AOB strains.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Eckert, C. A.; Sullivan, R.; Johnson, C.

CO2 and H2 are promising feedstocks for production of valuable biocompounds. Ralstonia eutropha utilizes these feedstocks to generate energy (ATP) and reductant (NAD(P)H) via oxidation of H2 by a membrane-bound (MBH) and a soluble hydrogenase (SH) for CO2 fixation by the Calvin-Benson-Bassham (CBB) cycle. Increased expression of the enzyme that fixes CO2 (RubisCO) resulted in 6-fold activity improvement in vitro, while increased expression of the MBH operon or the SH operon plus MBH operon maturation factors necessary for activity resulted in a 10-fold enhancement. Current research involves genetic manipulation of two endogenous cbb operons for increased expression, analysis of expressionmore » and activity of CBB/MBH/SH, cofactor ratios, and downstream products during autotrophic growth in control versus enhanced strains, and development of strategies for long-term, optimal overexpression. These studies will improve our understanding of autotrophic metabolism and provide a chassis strain for autotrophic production of biodiesel and other valuable carbon biocompounds.« less

Optimization of single-base-pair mismatch discrimination in oligonucleotide microarrays

NASA Technical Reports Server (NTRS)

Urakawa, Hidetoshi; El Fantroussi, Said; Smidt, Hauke; Smoot, James C.; Tribou, Erik H.; Kelly, John J.; Noble, Peter A.; Stahl, David A.

2003-01-01

The discrimination between perfect-match and single-base-pair-mismatched nucleic acid duplexes was investigated by using oligonucleotide DNA microarrays and nonequilibrium dissociation rates (melting profiles). DNA and RNA versions of two synthetic targets corresponding to the 16S rRNA sequences of Staphylococcus epidermidis (38 nucleotides) and Nitrosomonas eutropha (39 nucleotides) were hybridized to perfect-match probes (18-mer and 19-mer) and to a set of probes having all possible single-base-pair mismatches. The melting profiles of all probe-target duplexes were determined in parallel by using an imposed temperature step gradient. We derived an optimum wash temperature for each probe and target by using a simple formula to calculate a discrimination index for each temperature of the step gradient. This optimum corresponded to the output of an independent analysis using a customized neural network program. These results together provide an experimental and analytical framework for optimizing mismatch discrimination among all probes on a DNA microarray.

Application of nitrogen metabolism in autotrophic bacteria to chemosynthetic bioregeneration in space missions, supplement

NASA Technical Reports Server (NTRS)

Wixom, R. L.

1974-01-01

The chemolithotroph, Hydrogenomonas eutropha, was considered as a life support, bioregenerative system. This project focuses on several metabolic functions that are related to the proposed nitrogen cycle between man and this microbe. Specifically this organism has the capability to utilize as the sole nitrogen source such urine components as urea and fifteen individual amino acids, but not nine other amino acids. The effectiveness of utilization was high for many amino acids. Several specific growth inhibitions were also observed. The enzyme that catalyzes the incorporation of ammonia in the medium into amino acids was identified as a NADP-specific, L-glutamate dehydrogenase. This enzyme has a constitutive nature. This organism can synthesize all of its amino acids from carbon dioxide and ammonia. Therefore with the background literature of multiple pathways of individual amino acid biosyntheses, our evidence to date is consistent with the Hydrogeneomonas group having the same pathway of valine-isoleucine formation as the classical E. coli.

Phytomanagement of Cd-contaminated soils using maize (Zea mays L.) assisted by plant growth-promoting rhizobacteria.

PubMed

Moreira, Helena; Marques, Ana P G C; Franco, Albina R; Rangel, António O S S; Castro, Paula M L

2014-01-01

Zea mays (L.) is a crop widely cultivated throughout the world and can be considered suitable for phytomanagement due to its metal resistance and energetic value. In this study, the effect of two plant growth-promoting rhizobacteria, Ralstonia eutropha and Chryseobacterium humi, on growth and metal uptake of Z. mays plants in soils contaminated with up to 30 mg Cd kg(-1) was evaluated. Bacterial inoculation increased plant biomass up to 63% and led to a decrease of up to 81% in Cd shoot levels (4-88 mg Cd kg(-1)) and to an increase of up to 186% in accumulation in the roots (52-134 mg Cd kg(-1)). The rhizosphere community structure changed throughout the experiment and varied with different levels of Cd soil contamination, as revealed by molecular biology techniques. Z. mays plants inoculated with either of the tested strains may have potential application in a strategy of soil remediation, in particular short-term phytostabilization, coupled with biomass production for energy purposes.

Infrared thermography--a non-invasive tool to evaluate thermal status of neonatal pigs based on surface temperature.

PubMed

Kammersgaard, T S; Malmkvist, J; Pedersen, L J

2013-12-01

Hypothermia is a major cause of mortality in neonatal pigs. Infrared (IR) thermography is a promising non-invasive method to assess thermal status, but has not been evaluated for use on neonatal pigs from birth. The aim of this study was to evaluate the application of IR thermography as a non-invasive tool to estimate body temperature and assess the thermal status in newborn pigs by (1) estimating the relationship between surface temperature and rectal temperature (RT) in neonatal pigs; and (2) estimating the influence of air temperature (AT), birth weight and the time from birth on the relationship between surface temperature and RT. The method was evaluated on the basis of 1695 thermograms and 915 RTs on 91 neonatal pigs born in loose farrowing pens with floor heating at 34°C, and three different ATs (15°C, 20°C and 25°C). Full-body thermograms of the back and the side of the pigs and RT were acquired at 11 sampling times between birth and 48 h after birth. The maximum (IRmax), minimum, average of the full body and ear minimum IR surface temperatures were derived from the thermograms. IRmax had the highest correlation with RT (0.82) and was therefore used in the statistical analysis. The relation of RT by IRmax depended on time at: 0 h (slope: 0.20°C, P<0.001), 0.25 h (slope: 0.42°C, P<0.01), and 0.5 and 1 h after birth (slope: 0.68°C, P<0.001). After the 1st hour (1.5 to 48 h) the relation of RT by IRmax was no longer affected by time (slope: 0.63°C, P<0.001). The agreement between RT and IRmax was improved (P<0.001) after the 1st hour (RT-IRmax 0 to 1 h: 2.02 (1.44)°C; 1.5 to 48 h: 0.95 (0.85)°C). IRmax below 30°C was indicative of piglets having RT<32°C (91.3%). The location of IRmax was identified predominantly at the base of the ears (27/50), other sites in the region of the head (12/50) and the axilla area (8/50). There was a small but significant effect of the angle as IRmax_side-IRmax_back: mean 0.20°C (P<0.001). On the basis of the low difference between IRmax from back and side view thermograms, and the location of IRmax, the angle seems less important and thus the method has the potential to be used without the need for manual restraint of the pigs. On the basis of the results of this study, we propose that IRmax temperature from full-body thermograms has implication as a valid tool to assess the thermal status in neonatal piglets but not as an identical substitute for RT.

An Innovative Cloning Platform Enables Large-Scale Production and Maturation of an Oxygen-Tolerant [NiFe]-Hydrogenase from Cupriavidus necator in Escherichia coli

PubMed Central

Schiffels, Johannes; Pinkenburg, Olaf; Schelden, Maximilian; Aboulnaga, El-Hussiny A. A.; Baumann, Marcus E. M.; Selmer, Thorsten

2013-01-01

Expression of multiple heterologous genes in a dedicated host is a prerequisite for approaches in synthetic biology, spanning from the production of recombinant multiprotein complexes to the transfer of tailor-made metabolic pathways. Such attempts are often exacerbated, due in most cases to a lack of proper directional, robust and readily accessible genetic tools. Here, we introduce an innovative system for cloning and expression of multiple genes in Escherichia coli BL21 (DE3). Using the novel methodology, genes are equipped with individual promoters and terminators and subsequently assembled. The resulting multiple gene cassettes may either be placed in one vector or alternatively distributed among a set of compatible plasmids. We demonstrate the effectiveness of the developed tool by production and maturation of the NAD+reducing soluble [NiFe]-hydrogenase (SH) from Cupriavidus necator H16 (formerly Ralstonia eutropha H16) in E. coli BL21Star™ (DE3). The SH (encoded in hoxFUYHI) was successfully matured by co-expression of a dedicated set of auxiliary genes, comprising seven hyp genes (hypC1D1E1A2B2F2X) along with hoxW, which encodes a specific endopeptidase. Deletion of genes involved in SH maturation reduced maturation efficiency substantially. Further addition of hoxN1, encoding a high-affinity nickel permease from C. necator, considerably increased maturation efficiency in E. coli. Carefully balanced growth conditions enabled hydrogenase production at high cell-densities, scoring mg·(Liter culture)−1 yields of purified functional SH. Specific activities of up to 7.2±1.15 U·mg−1 were obtained in cell-free extracts, which is in the range of the highest activities ever determined in C. necator extracts. The recombinant enzyme was isolated in equal purity and stability as previously achieved with the native form, yielding ultrapure preparations with anaerobic specific activities of up to 230 U·mg−1. Owing to the combinatorial power exhibited by the presented cloning platform, the system might represent an important step towards new routes in synthetic biology. PMID:23861944

Identification of novel subgroup A variants with enhanced receptor binding and replicative capacity in primary isolates of anaemogenic strains of feline leukaemia virus.

PubMed

Stewart, Hazel; Adema, Karen W; McMonagle, Elizabeth L; Hosie, Margaret J; Willett, Brian J

2012-05-31

The development of anaemia in feline leukaemia virus (FeLV)-infected cats is associated with the emergence of a novel viral subgroup, FeLV-C. FeLV-C arises from the subgroup that is transmitted, FeLV-A, through alterations in the amino acid sequence of the receptor binding domain (RBD) of the envelope glycoprotein that result in a shift in the receptor usage and the cell tropism of the virus. The factors that influence the transition from subgroup A to subgroup C remain unclear, one possibility is that a selective pressure in the host drives the acquisition of mutations in the RBD, creating A/C intermediates with enhanced abilities to interact with the FeLV-C receptor, FLVCR. In order to understand further the emergence of FeLV-C in the infected cat, we examined primary isolates of FeLV-C for evidence of FeLV-A variants that bore mutations consistent with a gradual evolution from FeLV-A to FeLV-C. Within each isolate of FeLV-C, we identified variants that were ostensibly subgroup A by nucleic acid sequence comparisons, but which bore mutations in the RBD. One such mutation, N91D, was present in multiple isolates and when engineered into a molecular clone of the prototypic FeLV-A (Glasgow-1), enhanced replication was noted in feline cells. Expression of the N91D Env on murine leukaemia virus (MLV) pseudotypes enhanced viral entry mediated by the FeLV-A receptor THTR1 while soluble FeLV-A Env bearing the N91D mutation bound more efficiently to mouse or guinea pig cells bearing the FeLV-A and -C receptors. Long-term in vitro culture of variants bearing the N91D substitution in the presence of anti-FeLV gp70 antibodies did not result in the emergence of FeLV-C variants, suggesting that additional selective pressures in the infected cat may drive the subsequent evolution from subgroup A to subgroup C. Our data support a model in which variants of FeLV-A, bearing subtle differences in the RBD of Env, may be predisposed towards enhanced replication in vivo and subsequent conversion to FeLV-C. The selection pressures in vivo that drive the emergence of FeLV-C in a proportion of infected cats remain to be established.

Efficiency of a closed-coupled solar pasteurization system in treating roof harvested rainwater.

PubMed

Dobrowsky, P H; Carstens, M; De Villiers, J; Cloete, T E; Khan, W

2015-12-01

Many studies have concluded that roof harvested rainwater is susceptible to chemical and microbial contamination. The aim of the study was thus to conduct a preliminary investigation into the efficiency of a closed-coupled solar pasteurization system in reducing the microbiological load in harvested rainwater and to determine the change in chemical components after pasteurization. The temperature of the pasteurized tank water samples collected ranged from 55 to 57°C, 64 to 66°C, 72 to 74°C, 78 to 81°C and 90 to 91°C. Cations analyzed were within drinking water guidelines, with the exception of iron [195.59 μg/L (55°C)-170.1 μg/L (91°C)], aluminum [130.98 μg/L (78°C)], lead [12.81 μg/L (55°C)-13.2 μg/L (91°C)] and nickel [46.43 μg/L (55°C)-32.82 μg/L (78°C)], which were detected at levels above the respective guidelines in the pasteurized tank water samples. Indicator bacteria including, heterotrophic bacteria, Escherichia coli and total coliforms were reduced to below the detection limit at pasteurization temperatures of 72°C and above. However, with the use of molecular techniques Yersinia spp., Legionella spp. and Pseudomonas spp. were detected in tank water samples pasteurized at temperatures greater than 72°C. The viability of the bacteria detected in this study at the higher temperature ranges should thus be assessed before pasteurized harvested rainwater is used as a potable water source. In addition, it is recommended that the storage tank of the pasteurization system be constructed from an alternative material, other than stainless steel, in order for a closed-coupled pasteurization system to be implemented and produce large quantities of potable water from roof harvested rainwater. Copyright © 2015 Elsevier B.V. All rights reserved.

47 CFR 73.4091 - Direct broadcast satellites.

Code of Federal Regulations, 2010 CFR

2010-10-01

... Report and Order, General Docket 80-603, FCC 82-285, adopted June 23, 1982. 90 FCC 2d 676; 47 FR 31555, July 21, 1982. (b) See Memorandum Opinion and Order, FCC 82-427, adopted September 23, 1982. 91 FCC 2d. (c) See Memorandum Opinion and Order, FCC 82-498, adopted November 4, 1982. 91 FCC 2d. [48 FR 9012...

JPRS Report, Science & Technology, China

DTIC Science & Technology

1991-08-05

trima [more] Granulosis Virus [DTGV] and Its Characterization of Nuclear Acid and Protein [Yang Zhirong, Liu Shigui, et al.; SICHUAN DAXUE XUEBAO, No...KxC 6 o Superconductor Synthesized by Beijing University, CAS Institute of Physics [RENMIN RIBA, 17 Jul 91...Reviewed surement and metering systems, industrial production of 91FE0608 Beijing WULI [PHYSICS] in Chinese liquid hydrogen and long-term storage, long

Effect of Solution Treatment on Microstructure and Properties of Gd - AZ91 Magnesium Alloy

NASA Astrophysics Data System (ADS)

Li, Yao; Wang, Huiling

2018-01-01

In this paper, the Gd-AZ91 alloy was manufactured by adding rare earth element Gd in AZ91 magnesium alloy. The effects of solution treatment on the microstructures of rare earth elements Gd were investigated by means of optical microscopy, scanning electron microscopy, X-ray diffraction analysis and equipment for testing mechanical properties. The experimental results show that the addition of rare earth element Gd in AZ91 magnesium alloy can refine the alloy grain, turn β-Mg17Al12 phase into a discontinuous network or point structure, and produce granular compound Al2Gd in the alloy; when solution temperature is about 380 °C, the alloy structure is the best, the tensile strength of the alloy is the largest with the value larger than 250Mpa; when the solution temperature exceeds 380 °C, the alloy structure is coarsened and the mechanical properties of the alloy are reduced. With the increase of rare earth element Gd content, the tensile strength of the alloy shows a tendency to increase gradually, which Indicates that the addition of a certain amount of rare earth elements Gd can improve the plasticity of the alloy.

Genome-wide association study of esophageal squamous cell carcinoma in Chinese subjects identifies susceptibility loci at PLCE1 and C20orf54.

PubMed

Wang, Li-Dong; Zhou, Fu-You; Li, Xue-Min; Sun, Liang-Dan; Song, Xin; Jin, Yan; Li, Jiang-Man; Kong, Guo-Qiang; Qi, Hong; Cui, Juan; Zhang, Lian-Qun; Yang, Jie-Zhi; Li, Ji-Lin; Li, Xing-Chuan; Ren, Jing-Li; Liu, Zhi-Cai; Gao, Wen-Jun; Yuan, Ling; Wei, Wu; Zhang, Yan-Rui; Wang, Wei-Peng; Sheyhidin, Ilyar; Li, Feng; Chen, Bao-Ping; Ren, Shu-Wei; Liu, Bin; Li, Dan; Ku, Jian-Wei; Fan, Zong-Min; Zhou, Sheng-Li; Guo, Zhi-Gang; Zhao, Xue-Ke; Liu, Na; Ai, Yong-Hong; Shen, Fang-Fang; Cui, Wen-Yan; Song, Shuang; Guo, Tao; Huang, Jing; Yuan, Chao; Huang, Jia; Wu, Yue; Yue, Wen-Bin; Feng, Chang-Wei; Li, Hong-Lei; Wang, Yan; Tian, Jin-Ya; Lu, Yue; Yuan, Yi; Zhu, Wen-Liang; Liu, Min; Fu, Wen-Jing; Yang, Xia; Wang, Han-Jing; Han, Suo-Li; Chen, Jie; Han, Min; Wang, Hai-Yan; Zhang, Peng; Li, Xiu-Min; Dong, Jin-Cheng; Xing, Guo-Lan; Wang, Ran; Guo, Ming; Chang, Zhi-Wei; Liu, Hai-Lin; Guo, Li; Yuan, Zhi-Qing; Liu, Hai; Lu, Qin; Yang, Liu-Qin; Zhu, Fu-Guo; Yang, Xiu-Feng; Feng, Xiao-Shan; Wang, Zhou; Li, Yin; Gao, She-Gan; Qige, Qirenwang; Bai, Long-Tang; Yang, Wen-Jun; Lei, Guang-Yan; Shen, Zhong-Ying; Chen, Long-Qi; Li, En-Min; Xu, Li-Yan; Wu, Zhi-Yong; Cao, Wei-Ke; Wang, Jian-Po; Bao, Zhi-Qin; Chen, Ji-Li; Ding, Guang-Cheng; Zhuang, Xiang; Zhou, Ying-Fa; Zheng, Hou-Feng; Zhang, Zheng; Zuo, Xian-Bo; Dong, Zi-Ming; Fan, Dong-Mei; He, Xin; Wang, Jin; Zhou, Qi; Zhang, Qin-Xian; Jiao, Xin-Ying; Lian, Shi-Yong; Ji, Ai-Fang; Lu, Xiao-Mei; Wang, Jin-Sheng; Chang, Fu-Bao; Lu, Chang-Dong; Chen, Zhi-Guo; Miao, Jian-Jun; Fan, Zeng-Lin; Lin, Ruo-Bai; Liu, Tai-Jiang; Wei, Jin-Chang; Kong, Qing-Peng; Lan, Yu; Fan, Yu-Jing; Gao, Fu-Sheng; Wang, Tian-Yun; Xie, Dong; Chen, Shu-Qing; Yang, Wan-Cai; Hong, Jun-Yan; Wang, Liang; Qiu, Song-Liang; Cai, Zhi-Ming; Zhang, Xue-Jun

2010-09-01

We performed a genome-wide association study of esophageal squamous cell carcinoma (ESCC) by genotyping 1,077 individuals with ESCC and 1,733 control subjects of Chinese Han descent. We selected 18 promising SNPs for replication in an additional 7,673 cases of ESCC and 11,013 control subjects of Chinese Han descent and 303 cases of ESCC and 537 control subjects of Chinese Uygur-Kazakh descent. We identified two previously unknown susceptibility loci for ESCC: PLCE1 at 10q23 (P(Han combined for ESCC) = 7.46 x 10(-56), odds ratio (OR) = 1.43; P(Uygur-Kazakh for ESCC) = 5.70 x 10(-4), OR = 1.53) and C20orf54 at 20p13 (P(Han combined for ESCC) = 1.21 x 10(-11), OR = 0.86; P(Uygur-Kazakh for ESCC) = 7.88 x 10(-3), OR = 0.66). We also confirmed association in 2,766 cases of gastric cardia adenocarcinoma cases and the same 11,013 control subjects (PLCE1, P(Han for GCA) = 1.74 x 10(-39), OR = 1.55 and C20orf54, P(Han for GCA) = 3.02 x 10(-3), OR = 0.91). PLCE1 and C20orf54 have important biological implications for both ESCC and GCA. PLCE1 might regulate cell growth, differentiation, apoptosis and angiogenesis. C20orf54 is responsible for transporting riboflavin, and deficiency of riboflavin has been documented as a risk factor for ESCC and GCA.

Theoretical Crystal-Field Calculations for Rare-Earth Ions in III-V semiconductor Compounds

DTIC Science & Technology

1991-10-01

AD-A243 098 TIC HDL-TM-91-16 1 Ii! 1 I!EiIII ’ii F CT F October 1991 aC7 1991J Theoretical Crystal-Field Calculations for Rare-Earth Ions in III-V...0188). Washngton. DC 20503 1 . AGENCY USE ONLY (Leave bia*) 2. REPORT DATE 3. REPORT TYPE AND DATES COVERED October 1991 Summary, from Jan 91 toJul 91...Laboratories HDL-TM-9 1 -16 2800 Powder Mill Road Adelphi, MD 20783-1197 9. SPONSORNG#AONITORING AGENCY NAME(S) AND ADDRESS(ES) 10. SPONSORINGMONITORING

Thermus arciformis sp. nov., a thermophilic species from a geothermal area.

PubMed

Zhang, Xin-Qi; Ying, Yi; Ye, Ying; Xu, Xue-Wei; Zhu, Xu-Fen; Wu, Min

2010-04-01

Two aerobic, Gram-negative, non-motile, non-sporulating, yellow-pigmented bacteria, strains TH92(T) and TH91, were isolated from a hot spring located in Laibin, Guangxi, in the south-eastern geothermal area of China. The isolates grew at 40-77 degrees C (optimally at 70 degrees C) and at pH 6.0-9.5 (optimally at pH 7.5-8.0). Phylogenetic analysis of 16S rRNA gene sequences and levels of DNA-DNA relatedness together indicated that the new isolates represented a novel species of the genus Thermus with closest affinity to Thermus aquaticus, Thermus igniterrae and Thermus thermophilus. Compared with their closest relatives, strains TH92( T) and TH91 were able to assimilate a wider range of carbohydrates, amino acids and organic acids as sole carbon sources for growth, such as lactose and melibiose. The new isolates had lower combined levels of C(16 : 0 ) and iso-C(16 : 0) compared with their closest relatives. On the basis of polyphasic taxonomic characterization, strains TH92(T) and TH91 are considered to represent a single novel species of the genus Thermus, for which the name Thermus arciformis sp. nov. is proposed. The type strain is TH92(T) (=CGMCC 1.6992(T) =JCM 15153(T)).

32 CFR Appendix C to Part 1290 - Ticket Sample-A Moving Violation

Code of Federal Regulations, 2011 CFR

2011-07-01

... 32 National Defense 6 2011-07-01 2011-07-01 false Ticket Sample-A Moving Violation C Appendix C to Part 1290 National Defense Other Regulations Relating to National Defense DEFENSE LOGISTICS AGENCY.... 1290, App. C Appendix C to Part 1290—Ticket Sample—A Moving Violation EC25OC91.007 ...

32 CFR Appendix C to Part 1290 - Ticket Sample-A Moving Violation

Code of Federal Regulations, 2013 CFR

2013-07-01

... 32 National Defense 6 2013-07-01 2013-07-01 false Ticket Sample-A Moving Violation C Appendix C to Part 1290 National Defense Other Regulations Relating to National Defense DEFENSE LOGISTICS AGENCY.... 1290, App. C Appendix C to Part 1290—Ticket Sample—A Moving Violation EC25OC91.007 ...

[Comparison of methods for the identification of the most common yeasts in the clinical microbiology laboratory].

PubMed

Guelfand, L; Grisolía, P; Bozzano, C; Kaufman, S

2003-01-01

We evaluated different methods for the routine identification of medically important yeasts. A total of 150 clinical isolates: 25 C. albicans, 25 C. tropicalis, 25 C. glabrata, 25 C. parapsilosis, 8 C. guilliermondii, 11 C. krusei and 31 Cryptococcus neoformans were tested by Yeast Biochemical Card bioMerieux Vitek (YBC), CHROMagar Candida (CHR). The addition of yeast morphology in Corn Meal agar-Tween 80 (AM) to YBC and CHR was also evaluated. The reference methods used were: API 20C, germ tube formation, AM, Christensen urea and Birdseed agar. YBC identified 135 yeasts with an overall accuracy of 90%. Sensitivity (S) and specificity (E) were: 92-98% for C. albicans and C. tropicalis; 84-99% for C. papapsilosis; 100-99% for C. glabrata; 91-100% for C. krusei; 63-98% for C. guilliermondii and 90-99% for Cryptococcus neoformans, respectively. CHR identified correctly 100% for C. albicans, 92% for C. tropicalis and 91% for C. krusei. Both methods combined with AM provided 100% S and E. We found that YBC system was appropriate for identification of yeasts isolated from human sources. CHR was effective and easy to use for identification of C. albicans, C. tropicalis and C. krusei. The routine use of AM with both methods is recommended.

Experimental and DFT evaluation of the 1H and 13C NMR chemical shifts for calix[4]arenes

NASA Astrophysics Data System (ADS)

Guzzo, Rodrigo N.; Rezende, Michelle Jakeline Cunha; Kartnaller, Vinicius; Carneiro, José Walkimar de M.; Stoyanov, Stanislav R.; Costa, Leonardo Moreira da

2018-04-01

The density functional theory is employed to determine the efficiency of 11 exchange-correlation (XC) functionals to compute the 1H and 13C NMR chemical shifts of p-tert-butylcalix[4]arene (ptcx4, R1 = C(CH3)3) and congeners using the 6-31G(d,p) basis set. The statistical analysis shows that B3LYP, B3PW91 and PBE1PBE are the best XC functionals for the calculation of 1H chemical shifts. Moreover, the best results for the 13C chemical shifts are obtained using the LC-WPBE, M06-2X and wB97X-D functionals. The performance of these XC functionals is tested for three other calix[4]arenes: p-sulfonic acid calix[4]arene (sfxcx4 - R1 = SO3H), p-nitro-calix[4]arene (ncx4, R1 = NO2) and calix[4]arene (cx4 - R1 = H). For 1H chemical shifts B3LYP, B3PW91 and PBE1PBE yield similar results, although B3PW91 shows more consistency in the calculated error for the different structures. For 13C NMR chemical shifts, the XC functional that stood out as best is LC-WPBE. Indeed, the three functionals selected for each of 1H and 13C show good accuracy and can be used in future studies involving the prediction of 1H and 13C chemical shifts for this type of compounds.

Personal Health Risks Behaviour Profile among University Students in the South East Nigeria: Implication for Health Education

ERIC Educational Resources Information Center

Ilo, Cajetan I.; Onwunaka, Chinagorom; Nwimo, Ignatius O.

2015-01-01

This descriptive survey was carried out in order to determine the personal health risks behaviour profile among university students in the south east of Nigeria. A random sample of 900 students completed the questionnaire designed for the study. Out of this number 821, representing about 91.2% return rate, were used for data analysis. Means and…

Correlation of Specific Mutations in Line Probe Assay (LPA) and Drug Susceptibility Test (DST) with respect to Fluoroquinolone Resistance in Drug Resistant Mycobacterium tuberculosis

PubMed Central

Agrawal, Umang; Savaj, Pratik; Davda, Kanishka; Rodrigues, Camilla; Soman, Rajeev

2017-01-01

Abstract Background This study was done to investigate the utility of specific fluoroquinolone mutations in LPA in predicting the susceptibility in DST at WHO recommended Critical Concentrations of 0.5 and 2 µg/dL of moxifloxacin within a short time frame as provided by LPA. Methods In a retrospective study performed at a tertiary care hospital of Mumbai, India from October 2015 to February 2017, consecutive samples demonstrating fluoroquinolone resistance by LPA were selected. The LPA kit used was Hain Lifescience Genotype MTBDRsl (Version 1). It detects the following mutations in gyrA gene: MUT1: Ala90Val, MUT2: Ser91Pro, MUT3A: Asp94Ala, MUT3B: Asp94Asn/Tyr, MUT3C: Asp94Gly, MUT3D: Asp94His. The causal mutation was noted. For 89 of these samples, DST had been requested and results with Critical Concentration of 0.5µg/dL and 2µg/dL for moxifloxacin were available Results The 89 samples studied were as follows: Sputum (n = 60), paravertebral soft tissue (n = 2), bronchoalveolar fluid (n = 2), cerebrospinal fluid (n = 1), endotracheal tube secretion (n = 1), pleural fluid (n = 1) and site not recorded (22). 3 of these samples had double mutations. Results are as follows. Mutation in gyrA gene Number of samples (n) Susceptible at 0.5 µg/dL [n (%)] Susceptible at 2 µg/dL [n (%)] MUT1 (Ala90Val) 18 6(33.33) 16(88.89) MUT2 (Ser91Pro) 2 0(0) 1(50) MUT3A (Asp94Asn) 13 3(23.07) 11(84.61) MUT3B (Asp94Asn/Tyr) 6 1(16.67) 4(66.67) MUT3C (Asp94Gly) 51 4(8) 43(84.31) MUT3D (Asp94His) 2 0(0) 2(100) Conclusion This study showed a higher proportion of M. tuberculosis susceptibility at 2 µg/dL rather than at 0.5 µg/dL, to moxifloxacin for gyrA mutations Ala90Val (MUT1), Asp94Ala (MUT3A), Asp94Gly (MUT3C), Asp94His (MUT3D) but not for Ser91Pro (MUT2) and Asp94Asn/Tyr (MUT3B). However, the number of samples with Ser91Pro (MUT2) and Asp94Asn/Tyr (MUT3B) mutations was too small for meaningful conclusion. This susceptibility at a higher critical concentration of moxifloxacin may have clinical implications for use of high dose moxifloxacin. Since this information is available within a short time frame as provided by LPA, a more effective regimen could be devised 4 to 8 weeks earlier than after results of DST. This may result in faster sputum conversion and prevent amplification of resistance. Disclosures All authors: No reported disclosures.

Microstructure evolution and mechanical properties of nano-SiCp/AZ91 composite processed by extrusion and equal channel angular pressing (ECAP)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Qiao, X.G.; Ying, T.

Nano-SiCp/AZ91 magnesium matrix composite was fabricated by stir casting. The as-cast ingots were extruded at 350 °C, then processed by equal channel angular pressing (ECAP) at various temperatures (250 °C, 300 °C and 350 °C). Grains are significantly refined after the extrusion and the ECAP. A basal fibre texture was detected by neutron diffraction after the extrusion, which inclines about 45° to the extrusion direction (ED) after the ECAP. Nano-scaled SiC particles agglomerate in the as-cast composite. After the extrusion, the agglomeration tends to form continuous or discontinuous strips along the extrusion direction. By application of the ECAP, the agglomeratedmore » SiC particles are partly dispersed and the strips formed during the extrusion tend to be thinner and broken with the increasing pass number. The yield tensile strength (YTS) and the ultimate tensile strength (UTS) of the composite are dramatically increased after the extrusion. ECAP for one pass at various temperatures further increases the strength, however, the YTS decreases with the increasing ECAP temperature and the pass number. The Orowan equations predict the maximum YTS of the composite may be up to 400 MPa providing SiC particles are homogenously distributed in the matrix. - Highlights: •Nano-scaled SiC particles were successfully added into AZ91 by stirring casting. •Agglomeration of nano-particles were improved by extrusion and ECAP. •Yield strength of the composite is 328 MPa after one pass of ECAP. •Further ECAP process with optimized parameters may fully disperse nano-particles. •Yield strength is predicted to up to 400 MPa when particles are fully dispersed.« less

Automatic Author Profiling of Online Chat Logs

DTIC Science & Technology

2007-03-01

CLASSIFICATION WITH PRIOR ..........91 1. All Test Data ................................91 2. Extracted Test Data: Teens and 20s ...........92 3...Extracted Test Data: Teens and 30s ...........92 4. Extracted Test Data: Teens and 40s ...........93 5. Extracted Test Data: Teens and 50s ...........93 6...Data ................................97 C. AGE: BINARY CLASSIFICATION WITH PRIOR .............98 1. Extracted Test Data: Teens and 20s ...........98 2

7 CFR 3.91 - Adjusted civil monetary penalties.

Code of Federal Regulations, 2011 CFR

2011-01-01

... Secretary issued pursuant to the Dairy Product Mandatory Reporting program, codified at 7 U.S.C. 1637b(c)(4... Protection Act disqualification, codified at 15 U.S.C. 1825(c), has a maximum of $4,300. (x) Civil penalty....C. 1786(h)(8)(H)(i), has a maximum of $145,200,000. (v) Civil penalty for a vendor convicted of...

Extemporaneous sildenafil citrate oral suspensions for the treatment of pulmonary hypertension in children.

PubMed

Nahata, Milap C; Morosco, Richard S; Brady, Michael T

2006-02-01

The stability of sildenafil citrate 2.5 mg/mL in two extemporaneously prepared oral suspensions stored at 4 and 25 degrees C was studied. Thirty 25-mg tablets of sildenafil citrate were ground to powder, and the powder was combined with a 1:1 mixture of Ora-Sweet and Ora-Plus or a 1:1 mixture of methylcellulose 1% and Simple Syrup, NF, to produce two 2.5-mg/mL suspensions. Five plastic bottles of each suspension were stored in amber plastic prescription bottles at 4 or 25 degrees C. Samples were collected on days 0, 7, 14, 28, 42, 56, 70, and 91 for analysis of sildenafil content by high-performance liquid chromatography; pH was also measured. Samples were visually observed against black and white backgrounds. The mean concentration of sildenafil citrate exceeded 98% of the initial concentration in all samples at both temperatures throughout the 91-day study period. No changes in pH, odor, or physical appearance were observed. Sildenafil citrate 2.5 mg/mL in two extemporaneously compounded oral suspensions was stable for 91 days in plastic prescription bottles at 4 and 25 degrees C.

A multi-standard approach for GIAO (13)C NMR calculations.

PubMed

Sarotti, Ariel M; Pellegrinet, Silvina C

2009-10-02

The influence of the reference standard employed in the calculation of (13)C NMR chemical shifts was investigated over a large variety of known organic compounds, using different quantum chemistry methods and basis sets. After detailed analysis of the collected data, we found that methanol and benzene are excellent reference standards for computing NMR shifts of sp(3)- and sp-sp(2)-hybridized carbon atoms, respectively. This multi-standard approach (MSTD) performs better than TMS in terms of accuracy and precision and also displays much lower dependence on the level of theory employed. The use of mPW1PW91/6-31G(d)//mPW1PW91/6-31G(d) level is recommended for accurate (13)C NMR chemical shift prediction at low computational cost.

7 CFR 235.7 - Records and reports.

Code of Federal Regulations, 2012 CFR

2012-01-01

... surveys. (Sec. 14, Pub. L. 95-166, 91 Stat. 1338 (42 U.S.C. 1776); sec. 7, Pub. L. 95-627, 92 Stat. 3621 (42 U.S.C. 1776); 93 Stat. 837, Pub. L. 96-108 (42 U.S.C. 1776); secs. 804, 816, 817 and 819, Pub. L...

7 CFR 235.11 - Other provisions.

Code of Federal Regulations, 2014 CFR

2014-01-01

.... 14, Pub. L. 95-166, 91 Stat. 1338 (42 U.S.C. 1776); sec. 7, Pub. L. 95-627, 92 Stat. 3621 (42 U.S.C. 1776); secs. 805 and 819, Pub. L. 97-35, 95 Stat. 521-535 (42 U.S.C. 1773); sec. 7(a), Pub. L. 95-627...

7 CFR 235.7 - Records and reports.

Code of Federal Regulations, 2014 CFR

2014-01-01

... surveys. (Sec. 14, Pub. L. 95-166, 91 Stat. 1338 (42 U.S.C. 1776); sec. 7, Pub. L. 95-627, 92 Stat. 3621 (42 U.S.C. 1776); 93 Stat. 837, Pub. L. 96-108 (42 U.S.C. 1776); secs. 804, 816, 817 and 819, Pub. L...

7 CFR 235.11 - Other provisions.

Code of Federal Regulations, 2010 CFR

2010-01-01

.... 14, Pub. L. 95-166, 91 Stat. 1338 (42 U.S.C. 1776); sec. 7, Pub. L. 95-627, 92 Stat. 3621 (42 U.S.C. 1776); secs. 805 and 819, Pub. L. 97-35, 95 Stat. 521-535 (42 U.S.C. 1773); sec. 7(a), Pub. L. 95-627...

7 CFR 235.7 - Records and reports.

Code of Federal Regulations, 2010 CFR

2010-01-01

... surveys. (Sec. 14, Pub. L. 95-166, 91 Stat. 1338 (42 U.S.C. 1776); sec. 7, Pub. L. 95-627, 92 Stat. 3621 (42 U.S.C. 1776); 93 Stat. 837, Pub. L. 96-108 (42 U.S.C. 1776); secs. 804, 816, 817 and 819, Pub. L...

7 CFR 235.7 - Records and reports.

Code of Federal Regulations, 2013 CFR

2013-01-01

... surveys. (Sec. 14, Pub. L. 95-166, 91 Stat. 1338 (42 U.S.C. 1776); sec. 7, Pub. L. 95-627, 92 Stat. 3621 (42 U.S.C. 1776); 93 Stat. 837, Pub. L. 96-108 (42 U.S.C. 1776); secs. 804, 816, 817 and 819, Pub. L...

7 CFR 235.11 - Other provisions.

Code of Federal Regulations, 2012 CFR

2012-01-01

.... 14, Pub. L. 95-166, 91 Stat. 1338 (42 U.S.C. 1776); sec. 7, Pub. L. 95-627, 92 Stat. 3621 (42 U.S.C. 1776); secs. 805 and 819, Pub. L. 97-35, 95 Stat. 521-535 (42 U.S.C. 1773); sec. 7(a), Pub. L. 95-627...

7 CFR 235.11 - Other provisions.

Code of Federal Regulations, 2013 CFR

2013-01-01

.... 14, Pub. L. 95-166, 91 Stat. 1338 (42 U.S.C. 1776); sec. 7, Pub. L. 95-627, 92 Stat. 3621 (42 U.S.C. 1776); secs. 805 and 819, Pub. L. 97-35, 95 Stat. 521-535 (42 U.S.C. 1773); sec. 7(a), Pub. L. 95-627...

7 CFR 235.7 - Records and reports.

Code of Federal Regulations, 2011 CFR

2011-01-01

... surveys. (Sec. 14, Pub. L. 95-166, 91 Stat. 1338 (42 U.S.C. 1776); sec. 7, Pub. L. 95-627, 92 Stat. 3621 (42 U.S.C. 1776); 93 Stat. 837, Pub. L. 96-108 (42 U.S.C. 1776); secs. 804, 816, 817 and 819, Pub. L...

7 CFR 235.11 - Other provisions.

Code of Federal Regulations, 2011 CFR

2011-01-01

.... 14, Pub. L. 95-166, 91 Stat. 1338 (42 U.S.C. 1776); sec. 7, Pub. L. 95-627, 92 Stat. 3621 (42 U.S.C. 1776); secs. 805 and 819, Pub. L. 97-35, 95 Stat. 521-535 (42 U.S.C. 1773); sec. 7(a), Pub. L. 95-627...

Ionospheric Correction Using Two-Frequency Measurements

DTIC Science & Technology

1971-08-25

Instead of 385 km LCWi-I-l CENS3TY SA1EI IFAvlfCTCFY IS^.’JC flHZ PANCE 330«.92 3117.18 2929.60 27U2.37 2C.55.76 2370.11 2185-8...1734.22 1912.C9 2C93.19 2276.60 2461.68 2 647.91 2834.94 ;C22.45 3210.22 DELTA PANCE C.4036 C.4024 C.3979 C.3898 C.3781 C.3629 C.3444

40 CFR 91.312 - Analytical gases.

Code of Federal Regulations, 2014 CFR

2014-07-01

...) Purified nitrogen, also referred to as “zero-grade nitrogen” (Contamination≤1 ppm C, ≤1 ppm CO, ≤400 ppm... percent hydrogen, balance helium) (Contamination≤1 ppm C, ≤400 ppm CO) (4) Purified synthetic air, also... must be available: C3 H8 and purified synthetic air (dilute measurements); C3 H8 and purified nitrogen...

Catalytic glycerol steam reforming for hydrogen production

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dan, Monica, E-mail: monica.dan@itim-cj.ro; Mihet, Maria, E-mail: maria.mihet@itim-cj.ro; Lazar, Mihaela D., E-mail: diana.lazar@itim-cj.ro

2015-12-23

Hydrogen production from glycerol by steam reforming combine two major advantages: (i) using glycerol as raw material add value to this by product of bio-diesel production which is obtained in large quantities around the world and have a very limited utilization now, and (ii) by implication of water molecules in the reaction the efficiency of hydrogen generation is increased as each mol of glycerol produces 7 mol of H{sub 2}. In this work we present the results obtained in the process of steam reforming of glycerol on Ni/Al{sub 2}O{sub 3}. The catalyst was prepared by wet impregnation method and characterizedmore » through different methods: N{sub 2} adsorption-desorption, XRD, TPR. The catalytic study was performed in a stainless steel tubular reactor at atmospheric pressure by varying the reaction conditions: steam/carbon ratio (1-9), gas flow (35 ml/min -133 ml/min), temperature (450-650°C). The gaseous fraction of the reaction products contain: H{sub 2}, CH{sub 4}, CO, CO{sub 2}. The optimum reaction conditions as resulted from this study are: temperature 550°C, Gly:H{sub 2}O ratio 9:1 and Ar flow 133 ml/min. In these conditions the glycerol conversion to gaseous products was 43% and the hydrogen yield was 30%.« less

Differential DNA Methylation of MicroRNA Genes in Temporal Cortex from Alzheimer's Disease Individuals.

PubMed

Villela, Darine; Ramalho, Rodrigo F; Silva, Aderbal R T; Brentani, Helena; Suemoto, Claudia K; Pasqualucci, Carlos Augusto; Grinberg, Lea T; Krepischi, Ana C V; Rosenberg, Carla

2016-01-01

This study investigated for the first time the genomewide DNA methylation changes of noncoding RNA genes in the temporal cortex samples from individuals with Alzheimer's disease (AD). The methylome of 10 AD individuals and 10 age-matched controls were obtained using Illumina 450 K methylation array. A total of 2,095 among the 15,258 interrogated noncoding RNA CpG sites presented differential methylation, 161 of which were associated with miRNA genes. In particular, 10 miRNA CpG sites that were found to be hypermethylated in AD compared to control brains represent transcripts that have been previously associated with the disease. This miRNA set is predicted to target 33 coding genes from the neuregulin receptor complex (ErbB) signaling pathway, which is required for the neurons myelination process. For 6 of these miRNA genes (MIR9-1, MIR9-3, MIR181C, MIR124-1, MIR146B, and MIR451), the hypermethylation pattern is in agreement with previous results from literature that shows downregulation of miR-9, miR-181c, miR-124, miR-146b, and miR-451 in the AD brain. Our data implicate dysregulation of miRNA methylation as contributor to the pathogenesis of AD.

Symmetric Simple Map with Dipole Map for a Single-Null Divertor Tokamak

NASA Astrophysics Data System (ADS)

Ali, Halima; Watson, Michael; Punjabi, Alkesh; Boozer, Allen

1996-11-01

This investigation focuses on the effects of an externally placed dipole coil on the magnetic topology of a single-null divertor tokamak with a stochastic scrape-off layer using the Method of Maps (Punjabi A, Verma A and Boozer A, Phys Rev Lett), 69, 3322 (1992) and J Plasma Phys, 52, 91 (1994). The unperturbed magnetic topology is represented by the Symmetric Simple Map (Ali H, Watson M, Mayer C, Punjabi A and Boozer A, Bull Am Phys Soc), 40, 1855 (1995). The effect of dipole perturbation is repesented by the Dipole Map (Ali H, Watson M, Punjabi A and Boozer A, Sherwood Mtg), paper 1C20 (1996). A single dipole coil is placed across from the X-point below the last good surface. The strength of the dipole perturbation and the distance of the coil from the last good surface are varied. We observe that the dipole perturbation causes spatially intermittent chaos. This has significant implications for radiative divertor concepts as well for impurity control. We also present the detailed results on the effects of the dipole coil on the properties of the stochastic layer and the footprint of the field lines on the divertor plate. This work is supported by the US DOE OFES.

Deletion in a quantitative trait gene qPE9-1 associated with panicle erectness improves plant architecture during rice domestication.

PubMed

Zhou, Yong; Zhu, Jinyan; Li, Zhengyi; Yi, Chuandeng; Liu, Jun; Zhang, Honggen; Tang, Shuzhu; Gu, Minghong; Liang, Guohua

2009-09-01

Rice plant architecture is an important agronomic trait and a major determinant in high productivity. Panicle erectness is the preferred plant architecture in japonica rice, but the molecular mechanism underlying domestication of the erect panicle remains elusive. Here we report the map-based cloning of a major quantitative trait locus, qPE9-1, which plays an integral role in regulation of rice plant architecture including panicle erectness. The R6547 qPE9-1 gene encodes a 426-amino-acid protein, homologous to the keratin-associated protein 5-4 family. The gene is composed of three Von Willebrand factor type C domains, one transmembrane domain, and one 4-disulfide-core domain. Phenotypic comparisons of a set of near-isogenic lines and transgenic lines reveal that the functional allele (qPE9-1) results in drooping panicles, and the loss-of-function mutation (qpe9-1) leads to more erect panicles. In addition, the qPE9-1 locus regulates panicle and grain length, grain weight, and consequently grain yield. We propose that the panicle erectness trait resulted from a natural random loss-of-function mutation for the qPE9-1 gene and has subsequently been the target of artificial selection during japonica rice breeding.

Phyllanthus muellerianus and C6H15NO3 synergistic effects on 0.5 M H2SO4-immersed steel-reinforced concrete: Implication for clean corrosion-protection of wind energy structures in industrial environment

NASA Astrophysics Data System (ADS)

Okeniyi, Joshua Olusegun; Omotosho, Olugbenga Adeshola; Popoola, Abimbola Patricia Idowu; Loto, Cleophas Akintoye

2016-07-01

This paper investigates Phyllanthus muellerianus leaf-extract and C6H15NO3 (triethanolamine: TEA) synergistic effects on reinforcing-steel corrosion-inhibition and the compressive-strength of steel-reinforced concrete immersed in 0.5 M H2SO4. This is to assess suitability of the synergistic admixture usage for wind-energy steel-reinforced concrete structures designed for industrial environments. Steel-reinforced concrete specimens were admixed with individual and synergistic designs of Phyllanthus muellerianus leaf-extract and C6H15NO3 admixtures and immersed in the 0.5 M H2SO4. Electrochemical monitoring of corrosion potential, as per ASTM C876-91 R99, and corrosion current were obtained and statistically analysed, as per ASTM G16-95 R04, for modelling noise resistance. Post-immersion compressive-strength testing then followed, as per ASTM C39/C39M-03, for detailing the admixture effect on load-bearing strength of the steel-reinforced concrete specimens. Results showed that while individual Phyllanthus muellerianus leaf-extract concentrations exhibited better inhibition-efficiency performance than C6H15NO3, synergistic additions of C6H15NO3 to Phyllanthus muellerianus leaf-extract improved steel-rebar corrosion-inhibition. Thus, 6 g Phyllanthus muellerianus + 2 g C6H15NO3 synergistically improved inhibition-efficiency to η = 84.17%, from η = 55.28% by the optimal chemical or from η = 74.72% by the optimal plant-extract admixtures. The study also established that improved compressive strength of steel-reinforced concrete with acceptable inhibition of the steel-rebar corrosion could be attained through optimal combination of the Phyllanthus muellerianus leaf-extract and C6H15NO3 admixtures.

The Nature of Light: II. A Historical Survey from the Planck Era and Implications for Budding Physicists

ERIC Educational Resources Information Center

Oon, Pey Tee; Subramaniam, R.

2009-01-01

Following on from our previous article (Oon and Subramaniam 2009 "Phys. Educ." 44 384-91), here we trace ideas on the history of light from the Planck era to modern times. In particular, the seminal contributions of Planck, Einstein and de Broglie are highlighted. Some lesser known facets of the nature of light are also emphasized. It is stressed…

Source habitats for terrestrial vertebrates of focus in the interior Columbia basin: broadscale trends and management implications. Volume 1—Overview.

Treesearch

Michael J. Wisdom; Richard S. Holthausen; Barbara C. Wales; Christina D. Hargis; Victoria A. Saab; Danny C. Lee; Wendel J. Hann; Terrell D. Rich; Mary M. Rowland; Wally J. Murphy; Michelle R. Eames

2000-01-01

We defined habitat requirements (source habitats) and assessed trends in these habitats for 91 species of terrestrial vertebrates on 58 million ha (145 million acres) of public and private lands within the interior Columbia basin (hereafter referred to as the basin). We also summarized knowledge about species-road relations for each species and mapped source habitats...

DoD STINFO Manager Training Course STINFO Documentation

DTIC Science & Technology

2002-05-01

nawcws2.wsmr.army.mil PENNSYLVANIA ONR Manufacturing Technology Det Carderock Div NAVSURFWARCEN Attn: Philip M. Broudy (Code 20) John Bozewicz (Code 911...Available NTIS; 91N33013.) AD-A252 069 11 Pinelli, Thomas E.; Madeline Henderson; Ann P. Bishop; and Philip Doty. Chronology of Selected Literature...Mindy L Kotler . "Japanese Tech- nological Innovation: Implications for Large Commercial Aircraft and Knowledge Diffusion." Paper presented at the

47 CFR 73.160 - Vertical plane radiation characteristics, f(θ).

Code of Federal Regulations, 2013 CFR

2013-10-01

... significant figures shown here should not be interpreted as a limitation on the number of significant figures... electrical height: the sum of A and B; A+B. See Figure 1 of this section. EC01MR91.066 (3) For a... between H and A; H−A. See Figure 2 of this section. EC01MR91.067 (c) One of the above f(θ) formulas must...

47 CFR 73.160 - Vertical plane radiation characteristics, f(θ).

Code of Federal Regulations, 2014 CFR

2014-10-01

... significant figures shown here should not be interpreted as a limitation on the number of significant figures... electrical height: the sum of A and B; A+B. See Figure 1 of this section. EC01MR91.066 (3) For a... between H and A; H−A. See Figure 2 of this section. EC01MR91.067 (c) One of the above f(θ) formulas must...

47 CFR 73.160 - Vertical plane radiation characteristics, f(θ).

Code of Federal Regulations, 2012 CFR

2012-10-01

... significant figures shown here should not be interpreted as a limitation on the number of significant figures... electrical height: the sum of A and B; A+B. See Figure 1 of this section. EC01MR91.066 (3) For a... between H and A; H−A. See Figure 2 of this section. EC01MR91.067 (c) One of the above f(θ) formulas must...

Investigation of diffusional interaction between P91 grade ferritic steel and Fe-15 wt.%B alloy and study of kinetics of boride formation at high temperature

NASA Astrophysics Data System (ADS)

Rai, Arun Kumar; Vijayashanthi, N.; Tripathy, H.; Hajra, R. N.; Raju, S.; Murugesan, S.; Saroja, S.

2017-11-01

In the present study, the feasibility of employing the indigenously developed ferroboron alloy (Fe-15 wt.%B) as an alternate neutron shield material in combination with 9Cr-based ferritic steel (P91) clad in future Indian fast breeder reactors (FBR), has been investigated from a metallurgical perspective. Towards this goal, a series of diffusion couple experiments have been conducted at three different temperatures namely, 600, 700 and 800 °C for time durations up to 5000 h. The thickness of interaction layer has been monitored using standard metallographic procedures. The experiments revealed that ferroboron/P91 combination exhibited a tendency to form complex intermetallic borides at the interface. The structural and microstructural characterization of the interface confirmed that the reaction layer consists predominantly of borides of Fe and Cr of type FeB, Fe2B, (Fe,Cr)2B and (Fe,Cr)B. The measured variation of interaction layer thickness as a function of time and temperature have been modelled in terms of diffusion mediated interaction. The growth kinetics of borided layer has followed the parabolic law at each temperature, and the apparent activation energy for boride layer formation is found to be of the order of 115 kJ mol-1. This indicates that the kinetics of boriding could be governed by diffusion of B into the P91 matrix. Based on the findings of present study, an extrapolative estimate of the clad attack thickness at 550 °C for 60 years of operating time has been made and it turns out to be 210 ± 15 μm, which is less than the clad thickness of FBR shielding subassembly (4 mm) [1]. Thus, this study confirms that at testing temperatures from 550 to 600 °C, the ferroboron/P91 steel combination can be safely employed for shielding subassembly applications in fast reactors.

Impact of a Patient-Centered Medical Home on Access, Quality, and Cost

DTIC Science & Technology

2013-02-01

as annual hemoglobin A1c ( HbA1c ) testing with values greater than 9.0; low-density lipoprotein- cholesterol (LDL-C) values less than 100 mg/dL...Postimplementation HEDIS Measure Preperiod (%) Postperiod (%) HbA1c Test 84.5 91.8 HbA1c > 9.0 25.9 17.6 LDL Screening 78.2 88.6 LDL-C Value < 100 mg

Creep Deformation, Rupture Analysis, Heat Treatment and Residual Stress Measurement of Monolithic and Welded Grade 91 Steel for Power Plant Components

NASA Astrophysics Data System (ADS)

Shrestha, Triratna

Modified 9Cr-1 Mo (Grade 91) steel is currently considered as a candidate material for reactor pressure vessels (RPVs) and reactor internals for the Very High Temperature Reactor (VHTR), and in fossil-fuel fired power plants at higher temperatures and stresses. The tensile creep behavior of Grade 91 steel was studied in the temperature range of 600°C to 750°C and stresses between 35 MPa and 350 MPa. Heat treatment of Grade 91 steel was studied by normalizing and tempering the steel at various temperatures and times. Moreover, Thermo-Ca1c(TM) calculation was used to predict the precipitate stability and their evolution, and construct carbon isopleths of Grade 91 steel. Residual stress distribution across gas tungsten arc welds (GTAW) in Grade 91 steel was measured by the time-of-flight neutron diffraction using the Spectrometer for Materials Research at Temperature and Stress (SMARTS) diffractometer at Lujan Neutron Scattering Center, Los Alamos National Laboratory, Los Alamos, NM, USA. Analysis of creep results yielded stress exponents of ˜9-11 in the higher stress regime and ˜1 in the lower stress regime. The creep behavior of Grade 91 steel was described by the modified Bird-Mukherjee-Dorn relation. The rate-controlling creep deformation mechanism in the high stress regime was identified as the edge dislocation climb with a stress exponent of n = 5. On the other hand, the deformation mechanism in the Newtonian viscous creep regime (n = 1) was identified as the Nabarro-Herring creep. Creep rupture data were analyzed in terms of Monkman-Grant relation and Larson-Miller parameter. Creep damage tolerance factor and stress exponent were used to identify the cause of creep damage. The fracture surface morphology of the ruptured specimens was studied by scanning electron microscopy to elucidate the failure mechanisms. Fracture mechanism map for Grade 91 steel was developed based on the available material parameters and experimental observations. The microstructural evolution of heat treated steel was correlated with the differential scanning calorimetric study. The combination of microstructural studies with optical microscopy, scanning and transmission electron microscopy, microhardness profiles, and calorimetric plots helped in the understanding of the evolution of microstructure and precipitates in Grade 91 steel. The residual stresses were determined at the mid-thickness of the plate, 4.35 mm and 2.35 mm below the surface of the as-welded and post-weld heat treated plate. The residual stresses of the as-welded plate were compared with the post-weld heat treated plate. The post-weld heat treatment significantly reduced the residual stress in the base metal, heat affected zone, and the weld zone. Vickers microhardness profiles of the as-welded, and post-weld heat treated specimens were also determined and correlated with the observed residual stress profile and microstructure.

Effect of Long-Term Service on Microstructure and Mechanical Properties of Martensitic 9% Cr Steel

NASA Astrophysics Data System (ADS)

Golański, Grzegorz; Zielińska-Lipiec, Anna; Zieliński, Adam; Sroka, Marek

2017-03-01

The paper presents the results of research on the X10CrMoVNbN9-1 (T91) steel after long-term service. The material for testing was taken from a pipe section of a boiler superheater coil serviced for around 105,000 h at the temperature of 540 °C, at the pressure of 12.5 MPa. A quantitative analysis including the measurement of mean diameter of subgrains and precipitates as well as the density of dislocations of the examined steel was performed by means of TEM. The microscopic tests of T91 steel were complemented with the results of tests on mechanical properties which included also the short creep tests. After service, the investigated steel was characterized by a retained lath microstructure of tempered martensite with fine subgrain and quite large density of dislocations as well as numerous precipitates. In the microstructure, apart from the particles of M23C6 and MX (VX, NbC, V-wings), the precipitates of Laves phase and single particles of Z phase were revealed. It has been shown that the extent of degradation of the T91 steel microstructure was minor, which resulted from its low temperature of service. Performed tests of mechanical properties showed that these properties fulfilled the minimum requirements for this steel in the as-received condition. A favorable influence of fine precipitates of Laves phase on mechanical properties was observed. Moreover, an insignificant influence of single precipitates of Z phase on the creep resistance of the examined steel was stated.

28 CFR 76.1 - Purpose.

Code of Federal Regulations, 2010 CFR

2010-07-01

...-Drug Abuse Act of 1988 (the Act), 21 U.S.C. 844a. This part establishes procedures for imposing civil... CFR 1316.91(j)(2) in violation of 21 U.S.C. 844a and specifies the appeal rights of persons subject to...

Beneficial Effect of the Nitric Oxide Donor Compound 3-(1,3-Dioxoisoindolin-2-yl)Benzyl Nitrate on Dysregulated Phosphodiesterase 5, NADPH Oxidase, and Nitrosative Stress in the Sickle Cell Mouse Penis: Implication for Priapism Treatment.

PubMed

Silva, Fábio H; Karakus, Serkan; Musicki, Biljana; Matsui, Hotaka; Bivalacqua, Trinity J; Dos Santos, Jean L; Costa, Fernando F; Burnett, Arthur L

2016-11-01

Patients with sickle cell disease (SCD) display priapism, and dysregulated nitric oxide (NO) pathway may contribute to this condition. However, current therapies offered for the prevention of priapism in SCD are few. The 3-(1,3-dioxoisoindolin-2-yl)benzyl nitrate (compound 4C) was synthesized through molecular hybridization of hydroxyurea and thalidomide, which displays an NO-donor property. This study aimed to evaluate the effects of compound 4C on functional and molecular alterations of erectile function in murine models that display low NO bioavailability, SCD transgenic mice, and endothelial NO synthase and neuronal NO synthase double gene-deficient (dNOS -/ ) mice, focusing on the dysregulated NO-cGMP- phosphodiesterase type 5 (PDE5) pathway and oxidative stress in erectile tissue. Wild-type, SCD, and dNOS -/- mice were treated with compound 4C (100 μmol/kg/d, 3 weeks). Intracavernosal pressure in anesthetized mice was evaluated. Corpus cavernosum tissue was dissected free and mounted in organ baths. SCD and dNOS -/- mice displayed a priapism phenotype, which was reversed by compound 4C treatment. Increased corpus cavernosum relaxant responses to acetylcholine and electrical-field stimulation were reduced by 4C in SCD mice. Likewise, increased sodium nitroprusside-induced relaxant responses were reduced by 4C in cavernosal tissue from SCD and dNOS -/- mice. Compound 4C reversed PDE5 protein expression and reduced protein expressions of reactive oxygen species markers, NADPH oxidase subunit gp91 phox , and 3-nitrotyrosine in penises from SCD and dNOS -/- mice. In conclusion, 3-week therapy with the NO donor 4C reversed the priapism in murine models that display lower NO bioavailability. NO donor compounds may constitute an additional strategy to prevent priapism in SCD. Copyright © 2016 by The American Society for Pharmacology and Experimental Therapeutics.

Discovery of an artificial peptide agonist to the fibroblast growth factor receptor 1c/βKlotho complex from random peptide T7 phage display.

PubMed

Sakamoto, Kotaro; Kawata, Yayoi; Masuda, Yasushi; Umemoto, Tadashi; Ito, Takashi; Asami, Taiji; Takekawa, Shiro; Ohtaki, Tetsuya; Inooka, Hiroshi

2016-11-04

Fibroblast growth factor receptor-1c (FGFR1c)/βKlotho (KLB) complex is a receptor of fibroblast growth factor 21 (FGF21). Pharmacologically, FGF21 shows anti-obesity and anti-diabetic effects upon peripheral administration. Here, we report the development of an artificial peptide agonist to the FGFR1c/KLB heterodimer complex. The peptide, F91-8A07 (LPGRTCREYPDLWWVRCY), was discovered from random peptide T7 phage display and selectively bound to the FGFR1c/KLB complex, but not to FGFR1c and KLB individually. After subsequent peptide dimerization using a short polyethyleneglycol (PEG) linker, the dimeric F91-8A07 peptide showed higher potent agonist activity than that of FGF21 in cultured primary human adipocytes. Moreover, the dimeric peptide led to an expression of the early growth response protein-1 (Egr-1) mRNA in vivo, which is a target gene of FGFR1c. To the best of our knowledge, this is the first report of a FGFR1c/KLB complex-selective artificial peptide agonist. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

Nuclear resonance vibrational spectroscopy reveals the FeS cluster composition and active site vibrational properties of an O 2-tolerant NAD +-reducing [NiFe] hydrogenase

DOE PAGES

Lauterbach, Lars; Wang, Hongxin; Horch, Marius; ...

2014-10-30

Hydrogenases are complex metalloenzymes that catalyze the reversible splitting of molecular hydrogen into protons and electrons essentially without overpotential. The NAD+-reducing soluble hydrogenase (SH) from Ralstonia eutropha is capable of H 2 conversion even in the presence of usually toxic dioxygen. The molecular details of the underlying reactions are largely unknown, mainly because of limited knowledge of the structure and function of the various metal cofactors present in the enzyme. Here, all iron-containing cofactors of the SH were investigated by 57Fe specific nuclear resonance vibrational spectroscopy (NRVS). Our data provide experimental evidence for one [2Fe2S] center and four [4Fe4S] clusters,more » which is consistent with the amino acid sequence composition. Only the [2Fe2S] cluster and one of the four [4Fe4S] clusters were reduced upon incubation of the SH with NADH. This finding explains the discrepancy between the large number of FeS clusters and the small amount of FeS cluster-related signals as detected by electron paramagnetic resonance spectroscopic analysis of several NAD+-reducing hydrogenases. For the first time, Fe–CO and Fe–CN modes derived from the [NiFe] active site could be distinguished by NRVS through selective 13C labeling of the CO ligand. This strategy also revealed the molecular coordinates that dominate the individual Fe–CO modes. The present approach explores the complex vibrational signature of the Fe–S clusters and the hydrogenase active site, thereby showing that NRVS represents a powerful tool for the elucidation of complex biocatalysts containing multiple cofactors.« less

Nuclear resonance vibrational spectroscopy reveals the FeS cluster composition and active site vibrational properties of an O2-tolerant NAD+-reducing [NiFe] hydrogenase.

PubMed

Lauterbach, Lars; Wang, Hongxin; Horch, Marius; Gee, Leland B; Yoda, Yoshitaka; Tanaka, Yoshihito; Zebger, Ingo; Lenz, Oliver; Cramer, Stephen P

Hydrogenases are complex metalloenzymes that catalyze the reversible splitting of molecular hydrogen into protons and electrons essentially without overpotential. The NAD + -reducing soluble hydrogenase (SH) from Ralstonia eutropha is capable of H 2 conversion even in the presence of usually toxic dioxygen. The molecular details of the underlying reactions are largely unknown, mainly because of limited knowledge of the structure and function the various metal cofactors present in the enzyme. Here all iron-containing cofactors of the SH were investigated by 57 Fe specific nuclear resonance vibrational spectroscopy (NRVS). Our data provide experimental evidence for one [2Fe2S] center and four [4Fe4S] clusters, which is consistent with amino acid sequence composition. Only the [2Fe2S] cluster and one of the four [4Fe4S] clusters were reduced upon incubation of the SH with NADH. This finding explains the discrepancy between the large number of FeS clusters and the small amount of FeS cluster-related signals as detected by electron paramagnetic resonance spectroscopic analysis of several NAD + -reducing hydrogenases. For the first time, Fe-CO and Fe-CN modes derived from the [NiFe] active site could be distinguished by NRVS through selective 13 C labeling of the CO ligand. This strategy also revealed the molecular coordinates that dominate the individual Fe-CO modes. The present approach explores the complex vibrational signature of the Fe-S clusters and the hydrogenase active site, thereby showing that NRVS represents a powerful tool for the elucidation of complex biocatalysts containing multiple cofactors.

Enzymatic and spectroscopic properties of a thermostable [NiFe]‑hydrogenase performing H2-driven NAD+-reduction in the presence of O2.

PubMed

Preissler, Janina; Wahlefeld, Stefan; Lorent, Christian; Teutloff, Christian; Horch, Marius; Lauterbach, Lars; Cramer, Stephen P; Zebger, Ingo; Lenz, Oliver

2018-01-01

Biocatalysts that mediate the H 2 -dependent reduction of NAD + to NADH are attractive from both a fundamental and applied perspective. Here we present the first biochemical and spectroscopic characterization of an NAD + -reducing [NiFe]‑hydrogenase that sustains catalytic activity at high temperatures and in the presence of O 2 , which usually acts as an inhibitor. We isolated and sequenced the four structural genes, hoxFUYH, encoding the soluble NAD + -reducing [NiFe]‑hydrogenase (SH) from the thermophilic betaproteobacterium, Hydrogenophilus thermoluteolus TH-1 T (Ht). The HtSH was recombinantly overproduced in a hydrogenase-free mutant of the well-studied, H 2 -oxidizing betaproteobacterium Ralstonia eutropha H16 (Re). The enzyme was purified and characterized with various biochemical and spectroscopic techniques. Highest H 2 -mediated NAD + reduction activity was observed at 80°C and pH6.5, and catalytic activity was found to be sustained at low O 2 concentrations. Infrared spectroscopic analyses revealed a spectral pattern for as-isolated HtSH that is remarkably different from those of the closely related ReSH and other [NiFe]‑hydrogenases. This indicates an unusual configuration of the oxidized catalytic center in HtSH. Complementary electron paramagnetic resonance spectroscopic analyses revealed spectral signatures similar to related NAD + -reducing [NiFe]‑hydrogenases. This study lays the groundwork for structural and functional analyses of the HtSH as well as application of this enzyme for H 2 -driven cofactor recycling under oxic conditions at elevated temperatures. Copyright © 2017 Elsevier B.V. All rights reserved.

Escherichia coli NemA is an efficient chromate reductase that can be biologically immobilized to provide a cell free system for remediation of hexavalent chromium.

PubMed

Robins, Katherine J; Hooks, David O; Rehm, Bernd H A; Ackerley, David F

2013-01-01

Hexavalent chromium is a serious and widespread environmental pollutant. Although many bacteria have been identified that can transform highly water-soluble and toxic Cr(VI) to insoluble and relatively non-toxic Cr(III), bacterial bioremediation of Cr(VI) pollution is limited by a number of issues, in particular chromium toxicity to the remediating cells. To address this we sought to develop an immobilized enzymatic system for Cr(VI) remediation. To identify novel Cr(VI) reductase enzymes we first screened cell extracts from an Escherichia coli library of soluble oxidoreductases derived from a range of bacteria, but found that a number of these enzymes can reduce Cr(VI) indirectly, via redox intermediates present in the crude extracts. Instead, activity assays for 15 candidate enzymes purified as His6-tagged proteins identified E. coli NemA as a highly efficient Cr(VI) reductase (k(cat)/K(M)= 1.1×10(5) M(-1) s(-1) with NADH as cofactor). Fusion of nemA to the polyhydroxyalkanoate synthase gene phaC from Ralstonia eutropha enabled high-level biosynthesis of functionalized polyhydroxyalkanoate granules displaying stable and active NemA on their surface. When these granules were combined with either Bacillus subtilis glucose dehydrogenase or Candida boidinii formate dehydrogenase as a cofactor regenerating partner, high levels of chromate transformation were observed with only low initial concentrations of expensive NADH cofactor being required, the overall reaction being powered by consumption of the cheap sacrificial substrates glucose or formic acid, respectively. This system therefore offers promise as an economic solution for ex situ Cr(VI) remediation.

Escherichia coli NemA Is an Efficient Chromate Reductase That Can Be Biologically Immobilized to Provide a Cell Free System for Remediation of Hexavalent Chromium

PubMed Central

Robins, Katherine J.; Hooks, David O.; Rehm, Bernd H. A.; Ackerley, David F.

2013-01-01

Hexavalent chromium is a serious and widespread environmental pollutant. Although many bacteria have been identified that can transform highly water-soluble and toxic Cr(VI) to insoluble and relatively non-toxic Cr(III), bacterial bioremediation of Cr(VI) pollution is limited by a number of issues, in particular chromium toxicity to the remediating cells. To address this we sought to develop an immobilized enzymatic system for Cr(VI) remediation. To identify novel Cr(VI) reductase enzymes we first screened cell extracts from an Escherichia coli library of soluble oxidoreductases derived from a range of bacteria, but found that a number of these enzymes can reduce Cr(VI) indirectly, via redox intermediates present in the crude extracts. Instead, activity assays for 15 candidate enzymes purified as His6-tagged proteins identified E. coli NemA as a highly efficient Cr(VI) reductase (kcat/KM  = 1.1×105 M−1s−1 with NADH as cofactor). Fusion of nemA to the polyhydroxyalkanoate synthase gene phaC from Ralstonia eutropha enabled high-level biosynthesis of functionalized polyhydroxyalkanoate granules displaying stable and active NemA on their surface. When these granules were combined with either Bacillus subtilis glucose dehydrogenase or Candida boidinii formate dehydrogenase as a cofactor regenerating partner, high levels of chromate transformation were observed with only low initial concentrations of expensive NADH cofactor being required, the overall reaction being powered by consumption of the cheap sacrificial substrates glucose or formic acid, respectively. This system therefore offers promise as an economic solution for ex situ Cr(VI) remediation. PMID:23527133

Structural characteristics and elevated temperature mechanical properties of AJ62 Mg alloy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kubásek, J., E-mail: Jiri.Kubasek@vscht.cz; Vojtěch, D.; Martínek, M.

2013-12-15

Structure and mechanical properties of the novel casting AJ62 (Mg–6Al–2Sr) alloy developed for elevated temperature applications were studied. The AJ62 alloy was compared to commercial casting AZ91 (Mg–9Al–1Zn) and WE43 (Mg–4Y–3RE) alloys. The structure was examined by scanning electron microscopy, x-ray diffraction and energy dispersive spectrometry. Mechanical properties were characterized by Viskers hardness measurements in the as-cast state and after a long-term heat treatment at 250 °C/150 hours. Compressive mechanical tests were also carried out both at room and elevated temperatures. Compressive creep tests were conducted at a temperature of 250 °C and compressive stresses of 60, 100 and 140more » MPa. The structure of the AJ62 alloy consisted of primary α-Mg dendrites and interdendritic nework of the Al{sub 4}Sr and massive Al{sub 3}Mg{sub 13}Sr phases. By increasing the cooling rate during solidification from 10 and 120 K/s the average dendrite arm thickness decreased from 18 to 5 μm and the total volume fraction of the interdendritic phases from 20% to 30%. Both factors slightly increased hardness and compressive strength. The room temperature compressive strength and hardness of the alloy solidified at 30 K/s were 298 MPa and 50 HV 5, i.e. similar to those of the as-cast WE43 alloy and lower than those of the AZ91 alloy. At 250 °C the compressive strength of the AJ62 alloy decreased by 50 MPa, whereas those of the AZ91 and WE43 alloys by 100 and 20 MPa, respectively. The creep rate of the AJ62 alloy was higher than that of the WE43 alloy, but significantly lower in comparison with the AZ91 alloy. Different thermal stabilities of the alloys were discussed and related to structural changes during elevated temperature expositions. - Highlights: • Small effect of cooling rate on the compressive strength and hardness of AJ 62 • A bit lower compressive strength of AJ 62 compared to AZ91 at room temperature • Higher resistance of the AJ 62 alloy to the creep process in compression compared to AZ91 • Excellent thermal stability and creep resistance of the alloy WE 43 • Improved thermal stability and creep resistance in order WE43 > AJ62 >> AZ91.« less

Molecular diversity of HLA-Cw alleles in the Maratha community of Mumbai, Maharashtra, western India.

PubMed

Shankarkumar, U; Ghosh, K; Mohanty, D

2005-08-01

Recent advances suggest a significant role for the HLA-C locus as a target of alloreactions after bone marrow transplantation. The biological importance of products of the HLA-C locus, both as transplant antigens and as ligands for natural killer (NK) cells, is well established. A total of 10 different serologically defined HLA-Cw antigen specificities (Cw1-Cw10) are encoded by the C locus; however, there are now 151 different alleles that can be identified by molecular methods. Serological definition of Cw alleles therefore includes 20-50% blanks, which cannot be detected by the available antisera. We used the molecular method of polymerase chain reaction (PCR)-based sequence-specific amplification and probe hybridization to define Cw alleles in 91 individuals from the Maratha community, and compared the data with data for 92 serologically typed Maratha individuals from India. We identified Cw*12, Cw*14, Cw*15, Cw*16 and Cw*18, along with the serologically identified Cw*01, Cw*02, Cw*03, Cw*04, Cw*06 and Cw*07 alleles. The HLA-Cw blank allele frequency in the Maratha was reduced from 0.5706 to 0.00. Furthermore, by using a molecular technique, it was possible to identify novel allele subtypes, such as Cw*0104, Cw*0203 and Cw*0707, and a high frequency of Cw* 1801 in the Maratha community compared with other Indian and world populations. Our results will have clinical implications in related and unrelated HLA-matched bone marrow transplantation in India.

Strategic Implications of Intercommunal Warfare in Iraq

DTIC Science & Technology

2005-02-01

the militias associated with their home communities in a similar process as that of the disintegration of the Lebanese Army during Lebanon’s 1975-91...ancestral home and are deeply opposed to the idea that it may become a Kurdish city. U.S. statistics suggest that the Turkomans are only about 3 percent of ...go slow” approach to Kurdish political and cultural rights is replacing the more rigid policies of the past. This new

Bistable Vertical-Cavity Surface-Emitting Laser. Structures on GaAs and Si Substrates

DTIC Science & Technology

1994-06-01

vertical - cavity surface - emitting lasers ( VCSELs ) [1,5,6 of publications below], fabrication processes to realize low...May 91 through 1 June 94 R&T Number: Contract / Grant Number: N00014-91-J-1952 Contract / Grant Title: Bistable Vertical - Cavity Surface - Emitting Laser ...T.J. Rogers, B.G. Streetman, S.C. Smith, and R.D. Burnham, "Cascadabity of an Optically Iathing Vertical - Cavity Surface - Emitting Laser

Inhibition of proliferation and induction of apoptosis in soft tissue sarcoma cells by interferon-α and retinoids

PubMed Central

Brodowicz, T; Wiltschke, C; Kandioler-Eckersberger, D; Grunt, T W; Rudas, M; Schneider, S M; Hejna, M; Budinsky, A; Zielinski, C C

1999-01-01

Uncontrolled proliferation and a defect of apoptosis constitute crucial elements in the development and progression of tumours. Among many other biological response modifiers known to influence these mechanisms, the efficacy of retinoids and interferons in the treatment of various malignant entities is currently matter of discussion. In the present study, we have investigated the effects of 9-cis-retinoic acid (9cRA), 13-cis-retinoic acid (13cRA), all-trans-retinoic acid (tRA) and interferon-α on proliferation and apoptosis of human soft tissue sarcoma (STS) cell lines HTB-82 (rhabdomyosarcoma), HTB-91 (fibrosarcoma), HTB-92 (liposarcoma), HTB-93 (synovial sarcoma) and HTB-94 (chondrosarcoma) in relation to p53 genotype as well as p53 expression. HTB-91, HTB-92 and HTB-94 STS cells exhibited mutant p53, whereas wild-type p53 was found in HTB-93 STS cells, and a normal p53 status in HTB-82 STS cells, carrying a silent point mutation only. Interferon-α, irrespective of p53 status, inhibited the proliferation of all five cell lines dose- and time-dependently. Similarly, 9cRA, 13cRA and tRA decreased the proliferation of HTB-82 and HTB-93 STS cells, whereas the proliferation of p53-mutated HTB-91, HTB-92 and HTB-94 STS cells remained unchanged. Furthermore, only 9cRA and tRA were capable of inducing apoptosis in HTB-82 and HTB-93 STS cells, whereas HTB-91, HTB-92 and HTB-94 STS cells did not undergo apoptosis under the influence of 9cRA or tRA. Retinoic acid receptor (RAR)-α and RAR-β mRNA were not detectable by Northern blot analysis in the five STS cell lines, whereas mRNA for the universal retinoic acid receptor, RAR-γ, was expressed in all STS cell lines indicating that retinoid resistance was not associated with a lack of RAR expression. Apoptosis was not induced by interferon-α or 13cRA in any of the five STS cell lines tested. Our results indicate that within the panel of tested STS cell lines, inhibition of proliferation and induction of apoptosis result from different mechanisms which differ in their dependence upon the presence of intact p53. © 1999 Cancer Research Campaign PMID:10424735

7 CFR 3.91 - Adjusted civil monetary penalties.

Code of Federal Regulations, 2010 CFR

2010-01-01

... obey Horse Protection Act disqualification, codified at 15 U.S.C. 1825(c), has a maximum of $4,300. (x... bids for the purpose of influencing a bid, codified at 42 U.S.C. 1786(h)(8)(H)(i), has a maximum of... 42 U.S.C. 1786(o)(1)(A) and 42 U.S.C. 1786(o)(4)(B), has a maximum of $11,000 for each violation...

Mechanism of p53-Dependent Apoptosis and its Role in Breast Cancer Therapy

DTIC Science & Technology

1999-07-01

inducibly express p53 as previously described (Chen et a!., 1996). ( a ) Levels of p53, p21, and actin in p53-3, and p53(A62-91)-l, -5, and -6 cells...1994) was used as template, ( a ) Levels of p53, p21 and actin in p53-3 and p53(gln22-ser23/A62-91)-2 and -14 cells were assayed by Western blot...CGG TAC CCC TGT CAT CTT CTG TC; and reverse primer C393 as used for generating p53(A62-91). ( a ) Levels of p53, p21, and actin in p53-3, and p53(A74

Corrosion Behavior of AZ91D Magnesium Alloy in Three Different Physiological Environments

NASA Astrophysics Data System (ADS)

Zhou, Juncen; Li, Qing; Zhang, Haixiao; Chen, Funan

2014-01-01

Magnesium alloys have been considered as promising biomedical materials and were studied in different physiological environments. In this work, corrosion behavior of AZ91D magnesium alloy in artificial saliva, simulated body fluid (SBF), and 3.5 wt.% NaCl solution was investigated using electrochemical techniques and a short-term immersion test. In contrast with other physiological environments, the amount of aggressive ions in artificial saliva is small. In addition, a protective film is formed on the surface of samples in artificial saliva. Experimental results suggest that corrosion resistance of AZ91D magnesium alloy in artificial saliva is better than that in c-SBF and 3.5 wt.% NaCl solution.

Creep Rupture of the Simulated HAZ of T92 Steel Compared to that of a T91 Steel.

PubMed

Peng, Yu-Quan; Chen, Tai-Cheng; Chung, Tien-Jung; Jeng, Sheng-Long; Huang, Rong-Tan; Tsay, Leu-Wen

2017-02-08

The increased thermal efficiency of fossil power plants calls for the development of advanced creep-resistant alloy steels like T92. In this study, microstructures found in the heat-affected zone (HAZ) of a T92 steel weld were simulated to evaluate their creep-rupture-life at elevated temperatures. An infrared heating system was used to heat the samples to 860 °C (around A C1 ), 900 °C (slightly below A C3 ), and 940 °C (moderately above A C3 ) for one minute, before cooling to room temperature. The simulated specimens were then subjected to a conventional post-weld heat treatment (PWHT) at 750 °C for two hours, where both the 900 °C and 940 °C simulated specimens had fine grain sizes. In the as-treated condition, the 900 °C simulated specimen consisted of fine lath martensite, ferrite subgrains, and undissolved carbides, while residual carbides and fresh martensite were found in the 940 °C simulated specimen. The results of short-term creep tests indicated that the creep resistance of the 900 °C and 940 °C simulated specimens was poorer than that of the 860 °C simulated specimens and the base metal. Moreover, simulated T92 steel samples had higher creep strength than the T91 counterpart specimens.

Development of Genuine Neural Network Prototype Chip

DTIC Science & Technology

1991-01-28

priori distribution is equivalent, and more readily visualized with a rank curve . The sonar signal data consisted of approximately 85% class Target and...15% class Clutter. For this reason, the rank curves for the class Clutter were used for device parameter analysis. R & D STATUS REPORT 1/28/91 N00014...the signal CLASSLD#. Four 10-bit class probabilities are available on the output bus (C0-C9, C16-C25, C32-C41 and C48- C57 ) at each clock cycle. A

24 CFR 91.5 - Definitions.

Code of Federal Regulations, 2014 CFR

2014-04-01

...) of the Head Start Act (42 U.S.C. 9832(11)), section 41403(6) of the Violence Against Women Act of... Start Act (42 U.S.C. 9832), section 41403 of the Violence Against Women Act of 1994 (42 U.S.C. 14043e-2..., histories of domestic violence or childhood abuse (including neglect), the presence of a child or youth with...

24 CFR 91.5 - Definitions.

Code of Federal Regulations, 2012 CFR

2012-04-01

...) of the Head Start Act (42 U.S.C. 9832(11)), section 41403(6) of the Violence Against Women Act of... Start Act (42 U.S.C. 9832), section 41403 of the Violence Against Women Act of 1994 (42 U.S.C. 14043e-2..., histories of domestic violence or childhood abuse (including neglect), the presence of a child or youth with...

24 CFR 91.5 - Definitions.

Code of Federal Regulations, 2013 CFR

2013-04-01

...) of the Head Start Act (42 U.S.C. 9832(11)), section 41403(6) of the Violence Against Women Act of... Start Act (42 U.S.C. 9832), section 41403 of the Violence Against Women Act of 1994 (42 U.S.C. 14043e-2..., histories of domestic violence or childhood abuse (including neglect), the presence of a child or youth with...

Evidence for azimuthal variations of the oxygen-abundance gradient tracing the spiral structure of the galaxy HCG 91c

NASA Astrophysics Data System (ADS)

Vogt, F. P. A.; Pérez, E.; Dopita, M. A.; Verdes-Montenegro, L.; Borthakur, S.

2017-05-01

Context. The distribution of elements in galaxies forms an important diagnostic tool to characterize these systems' formation and evolution. This tool is, however, complex to use in practice, as galaxies are subject to a range of simultaneous physical processes active from pc to kpc scales. This renders observations of the full optical extent of galaxies down to sub-kpc scales essential. Aims: Using the WiFeS integral field spectrograph, we previously detected abrupt and localized variations in the gas-phase oxygen abundance of the spiral galaxy HCG 91c. Here, we follow-up on these observations to map HCG 91c's disk out to 2 Re at a resolution of 600 pc, and characterize the non-radial variations of the gas-phase oxygen abundance in the system. Methods: We obtained deep MUSE observations of the target under 0.6 arcsec seeing conditions. We perform both a spaxel-based and aperture-based analysis of the data to map the spatial variations of 12 +log (O/H) across the disk of the galaxy. Results: We confirm the presence of rapid variations of the oxygen abundance across the entire extent of the galaxy previously detected with WiFeS, for all azimuths and radii. The variations can be separated in two categories: a) localized and associated with individual H II regions; and b) extended over kpc scales, and occurring at the boundaries of the spiral structures in the galaxy. Conclusions: Our MUSE observations suggest that the enrichment of the interstellar medium in HGC 91c has proceeded preferentially along spiral structures, and less efficiently across them. Our dataset highlights the importance of distinguishing individual star-forming regions down to scales of a few 100 pc when using integral field spectrographs to spatially resolve the distribution of oxygen abundances in a given system, and accurately characterize azimuthal variations and intrinsic scatter. The movie associated to Fig. 8 is available at http://www.aanda.org

Power-law expansion of the Universe from the bosonic Lorentzian type IIB matrix model

NASA Astrophysics Data System (ADS)

Ito, Yuta; Nishimura, Jun; Tsuchiya, Asato

2015-11-01

Recent studies on the Lorentzian version of the type IIB matrix model show that (3+1)D expanding universe emerges dynamically from (9+1)D space-time predicted by superstring theory. Here we study a bosonic matrix model obtained by omitting the fermionic matrices. With the adopted simplification and the usage of a large-scale parallel computer, we are able to perform Monte Carlo calculations with matrix size up to N = 512, which is twenty times larger than that used previously for the studies of the original model. When the matrix size is larger than some critical value N c ≃ 110, we find that (3+1)D expanding universe emerges dynamically with a clear large- N scaling property. Furthermore, the observed increase of the spatial extent with time t at sufficiently late times is consistent with a power-law behavior t 1/2, which is reminiscent of the expanding behavior of the Friedmann-Robertson-Walker universe in the radiation dominated era. We discuss possible implications of this result on the original supersymmetric model including fermionic matrices.

Molecular cloning and expression profile analysis of porcine TCAP gene.

PubMed

Cheng, Hunjun; Xu, Xuewen; Zhao, Shuhong; Liu, Bang; Yu, Mei; Fan, Bin

2010-03-01

The gradually discovered sarcomeric proteins play important roles for structural integrity and signal transduction of sarcomere during myofibril genesis. TCAP (also described as telethonin, T-cap), one of the sarcomeric protein genes, is regulated developmentally. In this study, we reported the molecular characteristics of porcine TCAP gene. A 979 bp TCAP cDNA nucleotide sequence was obtained in pig and the deduced amino acid sequence had 92 and 91% identity to those of human and mouse homologous genes, respectively. One SNP was discovered and the allele frequency analysis showed that G allele frequency was low among 221 unrelated pigs from seven breeds. The tissue distribution patterns revealed that TCAP mRNA was expressed abundantly in skeletal and heart muscle tissue. Real-time quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) results displayed TCAP mRNA was up-regulated in both Tongcheng and Landrace pigs during prenatal skeletal muscle development stages. This study suggested that TCAP gene might be a prospective candidate gene affecting muscle mass and meat quality traits in the pig, and also implicated the possible significance of TCAP on sarcomere assembly.

Ab initio calculation of atomic interactions on Al(110): implications for epitaxial growth

NASA Astrophysics Data System (ADS)

Fichthorn, Kristen; Tiwary, Yogesh

2007-03-01

Using first-principles calculations based on density-functional theory, we resolved atomic interactions between adsorbed Al atoms on Al(110). Relevant pair and trio interactions were quantified. We find that pair interactions extend to the third in-channel and second cross-channel neighbor on the anisotropic (110) surface. Beyond these distances, pair interactions are negligible. The nearest-neighbor interaction in the in-channel direction is attractive, but nearest-neighbor cross-channel interaction is repulsive. While nearest-neighbor, cross-channel repulsion does not support the experimental observation of 3D hut formation in Al/Al(110) homoepitaxial growth [1], we find that trio interactions can be significant and attractive and they support cross-channel bonding. The pair and trio interactions have direct and indirect components. We have quantified the electronic and elastic components of the indirect, substrate-mediated interactions. We also probe the influence of these interactions on the energy barriers for adatom hopping. [1] F. Buatier de Mongeot, W. Zhu, A. Molle, R. Buzio, C. Boragno, U. Valbusa, E. Wang, and Z. Zhang, Phys. Rev. Lett. 91, 016102 (2003).

Simultaneous determination of fluoxetine and norfluoxetine enantiomers using isotope discrimination mass spectroscopy solution method and its application in the CYP2C9-mediated stereoselective interactions.

PubMed

Yu, Lushan; Wang, Shengjia; Jiang, Huidi; Zhou, Hui; Zeng, Su

2012-05-04

In this study, we developed an LC-MS/MS method based on an isotope discrimination mass spectroscopy solution (IDMSS) technology to simultaneously quantify enantiomers of fluoxetine (FLX) and norfluoxetine (NFLX) in a CYP2C9 incubation mixture. S-FLX and S-NFLX were labeled to form S-FLX-d5 and S-NFLX-d5. The method has several advantages over conventional chiral separation methods, in terms of the analysis period, resolution, and lower limit of quantification. The primary advantage of the method is that the two enantiomers can always be simultaneously determined by mass spectroscopy regardless if they are separated on column or not, owing to which it has high throughput and high sensitivity. The lower limit of quantification (amount on column) is 12.5 and 1.25 pg for FLX and NFLX, respectively. The retention time of FLX, NFLX, and the internal standard is only 1.9 min. The calibration curves were linear over the concentration range of 0.1-100 ng/ml for NFLX and 1-1000 ng/ml for FLX with an accepted reproducible (RSD<10%) and accurate (CV<10%). No significant kinetic isotope effect was found in the metabolism of S-FLX-d5 catalyzed by CYP2C9*1 and CYP2C9*2. The half-maximal inhibitory concentration values between R-FLX and S-FLX catalyzed by CYP2C9*1 and CYP2C9*2 were determined in this study. The inhibitory effects of R- to S-FLX were stronger than those of S- to R-FLX in both CYP2C9*1 and CYP2C9*2. The IDMSS technology is useful for stereoselective study of chiral compound in vitro. Copyright © 2012 Elsevier B.V. All rights reserved.

Phylogeny of fungal hemoglobins and expression analysis of the Aspergillus oryzae flavohemoglobin gene fhbA during hyphal growth.

PubMed

te Biesebeke, Rob; Levasseur, Anthony; Boussier, Amandine; Record, Eric; van den Hondel, Cees A M J J; Punt, Peter J

2010-01-01

The fhbA genes encoding putative flavohemoglobins (FHb) from Aspergillus niger and Aspergillus oryzae were isolated. Comparison of the deduced amino acid sequence of the A. niger fhbA gene and other putative filamentous fungal FHb-encoding genes to that of Ralstonia eutropha shows an overall conserved gene structure and completely conserved catalytic amino acids. Several yeasts and filamentous fungi, including both Aspergillus species have been found to contain a small FHb gene family mostly consisting of two family members. Based on these sequences the evolutionary history of the fungal FHb family was reconstructed. The isolated fhbA genes from A. oryzae and A. niger belong to a phylogenetic group, which exclusively contains Aspergillus genes. Different experimental approaches show that fhbA transcript levels appear during active hyphal growth. Moreover, in a pclA-disrupted strain with a hyperbranching growth phenotype, the transcript levels of the fhbA gene were 2–5 times higher compared to the wild-type. These results suggest that FHb from filamentous fungi have a function that is correlated to the hyphal growth phenotype.

Reversible [4Fe-3S] cluster morphing in an O(2)-tolerant [NiFe] hydrogenase.

PubMed

Frielingsdorf, Stefan; Fritsch, Johannes; Schmidt, Andrea; Hammer, Mathias; Löwenstein, Julia; Siebert, Elisabeth; Pelmenschikov, Vladimir; Jaenicke, Tina; Kalms, Jacqueline; Rippers, Yvonne; Lendzian, Friedhelm; Zebger, Ingo; Teutloff, Christian; Kaupp, Martin; Bittl, Robert; Hildebrandt, Peter; Friedrich, Bärbel; Lenz, Oliver; Scheerer, Patrick

2014-05-01

Hydrogenases catalyze the reversible oxidation of H(2) into protons and electrons and are usually readily inactivated by O(2). However, a subgroup of the [NiFe] hydrogenases, including the membrane-bound [NiFe] hydrogenase from Ralstonia eutropha, has evolved remarkable tolerance toward O(2) that enables their host organisms to utilize H(2) as an energy source at high O(2). This feature is crucially based on a unique six cysteine-coordinated [4Fe-3S] cluster located close to the catalytic center, whose properties were investigated in this study using a multidisciplinary approach. The [4Fe-3S] cluster undergoes redox-dependent reversible transformations, namely iron swapping between a sulfide and a peptide amide N. Moreover, our investigations unraveled the redox-dependent and reversible occurence of an oxygen ligand located at a different iron. This ligand is hydrogen bonded to a conserved histidine that is essential for H(2) oxidation at high O(2). We propose that these transformations, reminiscent of those of the P-cluster of nitrogenase, enable the consecutive transfer of two electrons within a physiological potential range.

A unique iron-sulfur cluster is crucial for oxygen tolerance of a [NiFe]-hydrogenase.

PubMed

Goris, Tobias; Wait, Annemarie F; Saggu, Miguel; Fritsch, Johannes; Heidary, Nina; Stein, Matthias; Zebger, Ingo; Lendzian, Friedhelm; Armstrong, Fraser A; Friedrich, Bärbel; Lenz, Oliver

2011-05-01

Hydrogenases are essential for H(2) cycling in microbial metabolism and serve as valuable blueprints for H(2)-based biotechnological applications. However, most hydrogenases are extremely oxygen sensitive and prone to inactivation by even traces of O(2). The O(2)-tolerant membrane-bound [NiFe]-hydrogenase of Ralstonia eutropha H16 is one of the few examples that can perform H(2) uptake in the presence of ambient O(2). Here we show that O(2) tolerance is crucially related to a modification of the internal electron-transfer chain. The iron-sulfur cluster proximal to the active site is surrounded by six instead of four conserved coordinating cysteines. Removal of the two additional cysteines alters the electronic structure of the proximal iron-sulfur cluster and renders the catalytic activity sensitive to O(2) as shown by physiological, biochemical, spectroscopic and electrochemical studies. The data indicate that the mechanism of O(2) tolerance relies on the reductive removal of oxygenic species guided by the unique architecture of the electron relay rather than a restricted access of O(2) to the active site.

Ammonia-oxidizing bacteria and archaea within biofilters of a commercial recirculating marine aquaculture system.

PubMed

Huang, Zhitao; Jiang, Yuli; Song, Xiefa; Hallerman, Eric; Peng, Lei; Dong, Dengpan; Ma, Teng; Zhai, Jieming; Li, Wensheng

2018-02-10

While biofilters are widely used to metabolize ammonia and other wastes in marine recirculating aquaculture systems, the ammonia-oxidizing bacterial and archaeal communities have not been characterized across a diversity of production systems. Using a metagenomics approach, we characterized the ammonia-oxidizing microbiological community of biofilters in a commercial recirculating marine aquaculture system producing hybrid grouper (Epinephelus lanceolatus × E. fuscoguttatus). Cloning and sequencing of the amoA gene showed that nitrifying bacteria included Nitrosomonas europea, N. stercoris, N. cryotolerans, N. eutropha, N. estuarii, eight strains of N. marina, and 15 strains not associated with described species. Nitrifying archaea included eight strains of Nitrosopumilus maritimus, N. koreensis, N. piranensis, N. adriaticus, undescribed congeners, and other undescribed archaea. The species composition of the bacterial and especially the archaeal communities was beyond that yet reported for aquaculture biofilters. While ammonia flux through the respective communities has yet to be estimated, the diverse environmental adaptations of the bacterial and archaeal communities suggest resilience of function under a range of environmental conditions.

Redox-controlled dinitrosyl formation at the diiron-oxo center of NorA.

PubMed

Cramm, Rainer; Strube, Katja

2008-01-01

In the denitrifying bacterium Ralstonia eutropha H16, the NorA protein is coproduced with the respiratory nitric oxide (NO) reductase. NorA contains a diiron-oxo center, which can form stable adducts with dioxygen and NO. In contrast to other diiron proteins, the formation of NorA-NO requires both fully reduced protein and additional electrons. A minor fraction of in vitro NorA-NO represents a paramagnetic dinitrosyl iron complex (DNIC), while the major fraction is attributed to a DNIC of the structure {Fe(NO)(2)}, which shows no electron paramagnetic resonance. The NorA-DNIC may be formed either upon direct reaction of the protein with NO or upon incubation with nitrite due to an intrinsic nitrite reduction activity of NorA that liberates NO. NorA can be purified rapidly as a six histidine-tagged derivative from overproducing cells of Escherichia coli. This chapter describes procedures for the preparation of different redox forms of NorA for the formation of NorA adducts with NO, dioxygen, and azide, as well as for the quantification of NorA-bound NO.

Microbial Surveillance of Potable Water Sources of the International Space Station

NASA Technical Reports Server (NTRS)

Bruce, Rebekah J.; Ott, C. Mark; Skuratov, Vladimir M.; Pierson, Duane L.

2005-01-01

To mitigate risk to the crew, the microbial surveillance of the quality of potable water sources of the International Space Station (ISS) has been ongoing since before the arrival of the first permanent crew. These water sources have included stored ground-supplied water, water produced by the shuttle fuel cells during flight, and ISS humidity condensate that is reclaimed and processed. Monitoring was accomplished using a self-contained filter designed to allow bacterial growth and enumeration during flight. Upon return to earth, microbial isolates were identified using 16S ribosomal gene sequencing. While the predominant isolates were common Gramnegative bacteria including Ralstonia eutropha, Methylobacterium fujisawaense, and Spingomonas paucimobilis, opportunistic pathogens such as Stenotrophomonas maltophilia and Pseudomonas aeruginosa were also isolated. Results of in-flight enumeration have indicated a fluctuation of bacterial counts above system design specifications. Additional in-flight monitoring capability for the specific detection of coliforms was added in 2004; no coliforms have been detected from any potable water source. Neither the bacterial concentrations nor the identification of the isolates recovered from these samples has suggested a threat to crew health.

A Bioluminescent Whole-Cell Reporter for Detection of 2,4-Dichlorophenoxyacetic Acid and 2,4-Dichlorophenol in Soil

PubMed Central

Hay, Anthony G.; Rice, James F.; Applegate, Bruce M.; Bright, Nathan G.; Sayler, Gary S.

2000-01-01

A bioreporter was made containing a tfdRPDII-luxCDABE fusion in a modified mini-Tn5 construct. When it was introduced into the chromosome of Ralstonia eutropha JMP134, the resulting strain, JMP134-32, produced a sensitive bioluminescent response to 2,4-dichlorophenoxyacetic acid (2,4-D) at concentrations of 2.0 μM to 5.0 mM. This response was linear (R2 = 0.9825) in the range of 2.0 μM to 1.1 × 102 μM. Saturation occurred at higher concentrations, with maximal bioluminescence occurring in the presence of approximately 1.2 mM 2,4-D. A sensitive response was also recorded in the presence of 2,4-dichlorophenol at concentrations below 1.1 × 102 μM; however, only a limited bioluminescent response was recorded in the presence of 3-chlorobenzoic acid at concentrations below 1.0 mM. A significant bioluminescent response was also recorded when strain JMP134-32 was incubated with soils containing aged 2,4-D residues. PMID:11010925

Prevalence of Nitrosomonas cluster 7 populations in the ammonia-oxidizing community of a submerged membrane bioreactor treating urban wastewater under different operation conditions.

PubMed

Cerrone, F; Poyatos, J M; Molina-Muñoz, M; Cortés-Lorenzo, C; González-López, J; Rodelas, B

2013-07-01

A pilot-scale ultrafiltration membrane bioreactor (MBR) was used for the aerobic treatment of urban wastewater in four experimental stages influenced by seasonal temperature and different sets of operation conditions. The structure of the ammonia-oxidizing bacteria (AOB) community was profiled by temperature gradient gel electrophoresis (TGGE), based on the amplification and separation of partial ammonia-monoxygenase subunit A (amoA) genes. Canonical correspondence analysis revealed that temperature, hydraulic retention time and percentage of ammonia removal had a significant effect on the fingerprints of AOB communities. Phylogenetic analysis conducted on amoA/AmoA sequences of reamplified TGGE bands showed, however, that closely related ammonia-oxidizing populations inhabited the sludge of the MBR in all experimental stages. Nitrosomonas cluster 7 populations (N. europaea-N. eutropha cluster) prevailed under all conditions tested, even when the MBR was operated under complete biomass retention or at low temperatures, suggesting that the high ammonia concentrations in the system were determinant to select r-strategist AOB.

A forward-design approach to increase the production of poly-3-hydroxybutyrate in genetically engineered Escherichia coli.

PubMed

Kelwick, Richard; Kopniczky, Margarita; Bower, Iain; Chi, Wenqiang; Chin, Matthew Ho Wai; Fan, Sisi; Pilcher, Jemma; Strutt, James; Webb, Alexander J; Jensen, Kirsten; Stan, Guy-Bart; Kitney, Richard; Freemont, Paul

2015-01-01

Biopolymers, such as poly-3-hydroxybutyrate (P(3HB)) are produced as a carbon store in an array of organisms and exhibit characteristics which are similar to oil-derived plastics, yet have the added advantages of biodegradability and biocompatibility. Despite these advantages, P(3HB) production is currently more expensive than the production of oil-derived plastics, and therefore, more efficient P(3HB) production processes would be desirable. In this study, we describe the model-guided design and experimental validation of several engineered P(3HB) producing operons. In particular, we describe the characterization of a hybrid phaCAB operon that consists of a dual promoter (native and J23104) and RBS (native and B0034) design. P(3HB) production at 24 h was around six-fold higher in hybrid phaCAB engineered Escherichia coli in comparison to E. coli engineered with the native phaCAB operon from Ralstonia eutropha H16. Additionally, we describe the utilization of non-recyclable waste as a low-cost carbon source for the production of P(3HB).

A Forward-Design Approach to Increase the Production of Poly-3-Hydroxybutyrate in Genetically Engineered Escherichia coli

PubMed Central

Kelwick, Richard; Kopniczky, Margarita; Bower, Iain; Chi, Wenqiang; Chin, Matthew Ho Wai; Fan, Sisi; Pilcher, Jemma; Strutt, James; Webb, Alexander J.; Jensen, Kirsten; Stan, Guy-Bart; Kitney, Richard; Freemont, Paul

2015-01-01

Biopolymers, such as poly-3-hydroxybutyrate (P(3HB)) are produced as a carbon store in an array of organisms and exhibit characteristics which are similar to oil-derived plastics, yet have the added advantages of biodegradability and biocompatibility. Despite these advantages, P(3HB) production is currently more expensive than the production of oil-derived plastics, and therefore, more efficient P(3HB) production processes would be desirable. In this study, we describe the model-guided design and experimental validation of several engineered P(3HB) producing operons. In particular, we describe the characterization of a hybrid phaCAB operon that consists of a dual promoter (native and J23104) and RBS (native and B0034) design. P(3HB) production at 24 h was around six-fold higher in hybrid phaCAB engineered Escherichia coli in comparison to E. coli engineered with the native phaCAB operon from Ralstonia eutropha H16. Additionally, we describe the utilization of non-recyclable waste as a low-cost carbon source for the production of P(3HB). PMID:25699671

RNAi validation of resistance genes and their interactions in the highly DDT-resistant 91-R strain of Drosophila melanogaster.

PubMed

Gellatly, Kyle J; Yoon, Kyong Sup; Doherty, Jeffery J; Sun, Weilin; Pittendrigh, Barry R; Clark, J Marshall

2015-06-01

4,4'-dichlorodiphenyltrichloroethane (DDT) has been re-recommended by the World Health Organization for malaria mosquito control. Previous DDT use has resulted in resistance, and with continued use resistance will increase in terms of level and extent. Drosophila melanogaster is a model dipteran that has many available genetic tools, numerous studies done on insecticide resistance mechanisms, and is related to malaria mosquitoes allowing for extrapolation. The 91-R strain of D. melanogaster is highly resistant to DDT (>1500-fold), however, there is no mechanistic scheme that accounts for this level of resistance. Recently, reduced penetration, increased detoxification, and direct excretion have been identified as resistance mechanisms in the 91-R strain. Their interactions, however, remain unclear. Use of UAS-RNAi transgenic lines of D. melanogaster allowed for the targeted knockdown of genes putatively involved in DDT resistance and has validated the role of several cuticular proteins (Cyp4g1 and Lcp1), cytochrome P450 monooxygenases (Cyp6g1 and Cyp12d1), and ATP binding cassette transporters (Mdr50, Mdr65, and Mrp1) involved in DDT resistance. Further, increased sensitivity to DDT in the 91-R strain after intra-abdominal dsRNA injection for Mdr50, Mdr65, and Mrp1 was determined by a DDT contact bioassay, directly implicating these genes in DDT efflux and resistance. Copyright © 2015 Elsevier Inc. All rights reserved.

Thermo-Mechanical Fatigue of a Fiber Reinforced Titanium Aluminide Composite

DTIC Science & Technology

1991-03-01

Puiitr]on Air Foic. Base, Oiio 91 10 25 038 DTIC 4E L L E C :i<. THERMO- MECHAN I CAL FAT I GUE OF A FIBER REINFORCED TITANIUM ALUMINIDE COMPOSITE Thesis...reinforced titanium aluminide composite was investigated. Three test specimens were subjected to in-phase thermo- mechanical cycling and four to out-of...Unclassified UL ............................... AFIT/GAE/ENY/91S-1 THERMO- MECHANICAL FATIGUE OF A FIBER REINFORCED TITANIUM ALUMINIDE COMPOSITE THESIS

Word Criticality Analysis. MOS: 67U. Skill Levels 1 & 2.

DTIC Science & Technology

1981-09-01

8217C~E 1651 47p 2 i Pr’ k 9901 22. u’ lt.I V% p 1 ’.2.2 125.1 I SC 0 1dept OW Met33, 2 5I2?.~ Viol 2 s:f STT3" 1601 2 S,1 -"𔃾 )vp 3 2 S F3.L I,,# I Io...97,1 91.1 08. 2 50cp? 14603 132 1 54i 1 6.2 156, P 3 C ,’TICAL 50 I’ll 20.4 8.5 󈨋 04 19𔃼 11.3 16,9 9,1 3 C f"IC .L - AGENT 1(),3 3 Cf...stal Viol , 94.1 ?)#1 129,1 lir7*2 llb0a ? Woo 1#1 1540.1 15301 120.1 150.1 173.1 11201 83.1 $)at $301 105.1 1,34,1, Ilil 1601 les 17L01U7151~ 1 7001

Tables of Queue Size and Waiting Time Distributions for M/M/c, M/D/c, and D/M/c Queueing Systems.

DTIC Science & Technology

1980-03-01

1091.0se-G. 4.11,61148 III- U .3 3"’s .2% .... 1.. 00-. :=1 2"’ all, I w 󈧕". 1 6.11:,""o 209.4 s. of 0 ::::v Iso ".. ...... t. 1.0 it oj .1. .11211111-41...72204011-06 A.0. " : " : 3:: 2;.1000561-02 .1.26 It G. 10 02 .40.. 1 a 2, ISO taOj *::% 9,110- 1 20 1. 11-.; 0-04161. 1. 1, 11" I-SP It-01 0.1.1002 21...0. 󈧄 .4 . . ISO 91-0) In* 66: : 0 14.12 b"ll: 00- 0.1.91ij 1 0 1 1: ?1 1, -"i’ll-02 10 1 1112 .2.0 1 0.99-8 1299-05 0. 9VOSIS 1 2. -. 2 .10 . - 0

Corrosion Behavior of Steels in Supercritical CO 2 for Power Cycle Applications

DOE Office of Scientific and Technical Information (OSTI.GOV)

Repukaiti, Richard; Teeter, Lucas; Ziomek-Moroz, Margaret

In order to understand issues with corrosion of heat exchanger materials in direct supercritical carbon dioxide (sCO 2) power cycles, a series of autoclave exposure experiments and electrochemical experiments have been conducted. Corrosion behaviors of 347H stainless steel and P91 martensitic-ferrtic steel in sCO 2 environment have been compared. In autoclave exposure tests performed at 50°C- 245°C and 80 bar. Mass change measurements, surface characterization, and corrosion product analysis have been conducted to understand the corrosion behavior of steels in sCO 2 containing H 2O and O 2. Electrochemical tests performed at room temperature and 50°C, a simulation environment ofmore » water condensation phase with dissolved CO 2 was prepared to evaluate the corrosion resistance of materials. From both types of experiments, generally 347H showed higher corrosion resistance than P91.« less

Availability-Based Importance Framework for Supplier Selection

DTIC Science & Technology

2015-04-30

IMA Journal of Management Math, 15(2), 161– 174. Chen, C . -T., Lin, C . -T., & Huang, S. -F. (2006). A fuzzy approach for supplier evaluation and...reliability modeling: Principles and applications. Hoboken, NJ: Wiley. Liao, C . -N., & Kao, H. -P. (2011). An integrated fuzzy TOPSIS and MCGP approach to...5307–5326. Wang, J. -W., Cheng, C . -H., & Huang, K.- C . (2009). Fuzzy hierarchical TOPSIS for supplier selection. Applied Soft Computing, 9(1), 377

Functional and biochemical properties of Mal de Río Cuarto virus (Fijivirus, Reoviridae) P9-1 viroplasm protein show further similarities to animal reovirus counterparts.

PubMed

Maroniche, Guillermo A; Mongelli, Vanesa C; Peralta, Andrea V; Distéfano, Ana J; Llauger, Gabriela; Taboga, Oscar A; Hopp, Esteban H; del Vas, Mariana

2010-09-01

Mal de Río Cuarto virus (MRCV) is a plant virus of the genus Fijivirus within the family Reoviridae that infects several monocotyledonous species and is transmitted by planthoppers in a persistent and propagative manner. Other members of the family replicate in viral inclusion bodies (VIBs) termed viroplasms that are formed in the cytoplasm of infected plant and insect cells. In this study, the protein coded by the first ORF of MRCV segment S9 (P9-1) was shown to establish cytoplasmic inclusion bodies resembling viroplasms after transfection of Spodoptera frugiperda insect cells. In accordance, MRCV P9-1 self-associates giving rise to high molecular weight complexes when expressed in bacteria. Strong self-interaction was also evidenced by yeast two-hybrid assays. Furthermore, biochemical characterization showed that MRCV P9-1 bound single stranded RNA and had ATPase activity. Finally, the MRCV P9-1 region required for the formation of VIB-like structures was mapped to the protein carboxy-terminal half. This extensive functional and biochemical characterization of MRCV P9-1 revealed further similarities between plant and animal reovirus viroplasm proteins. Copyright (c) 2010 Elsevier B.V. All rights reserved.

Development of analyses by high-performance liquid chromatography and liquid chromatography/tandem mass spectrometry of bilberry (Vaccinium myrtilus) anthocyanins in human plasma and urine.

PubMed

Cooke, Darren N; Thomasset, Sarah; Boocock, David J; Schwarz, Michael; Winterhalter, Peter; Steward, William P; Gescher, Andreas J; Marczylo, Timothy H

2006-09-20

Anthocyanins are potent antioxidants that may possess chronic disease preventive properties. Here, rapid, reliable, and reproducible solid-phase extraction, high-performance liquid chromatography (HPLC), and mass spectrometry techniques are described for the isolation, separation, and identification of anthocyanins in human plasma and urine. Recoveries of cyanidin-3-glucoside (C3G) were 91% from water, 71% from plasma, and 81% from urine. Intra- and interday variations for C3G extraction were 9 and 9.1% in plasma and 7.1 and 9.1% in urine and were less than 15% for all anthocyanins from a standardized bilberry extract (mirtoselect). Analysis of mirtoselect by HPLC with UV detection produced spectra with 15 peaks compatible with anthocyanin components found in mirtoselect within a total run time of 15 min. Chromatographic analysis of human urine obtained after an oral dose of mirtoselect yielded 19 anthocyanin peaks. Mass spectrometric analysis employing multiple reaction monitoring suggests the presence of unchanged anthocyanins and anthocyanidin glucuronide metabolites.

40 CFR 52.2027 - Approval status of Pennsylvania's Generic NOX and VOC RACT Rules.

Code of Federal Regulations, 2012 CFR

2012-07-01

... through 129.95 (see § 52.2020 (c)(129)) as those regulations apply to the Pittsburgh-Beaver Valley area... its approval of 25 PA Code of Regulations, Chapter 129.91 through 129.95 [see § 52.2020 (c)(129)] as...

40 CFR 52.2027 - Approval status of Pennsylvania's Generic NOX and VOC RACT Rules.

Code of Federal Regulations, 2013 CFR

2013-07-01

... through 129.95 (see § 52.2020 (c)(129)) as those regulations apply to the Pittsburgh-Beaver Valley area... its approval of 25 PA Code of Regulations, Chapter 129.91 through 129.95 [see § 52.2020 (c)(129)] as...

Brigade Intelligence Operations. Implications for the Nonlinear Battlefield

DTIC Science & Technology

1990-11-21

Intellingence School of Advanced Military Studies United States Army Command and General Staff College Fort Leavenworth2 Kansas~~First Term 90-91 / Approved...has few equipment needs. The section has an M577 staff track and a wheeled vehicle for transportion and communications. It acts as the net control... vehicle for operating on the move or for monitoring the brigade command net. There is no intelligence specific equipment assigned to the S2 section

Creep Rupture of the Simulated HAZ of T92 Steel Compared to that of a T91 Steel

PubMed Central

Peng, Yu-Quan; Chen, Tai-Cheng; Chung, Tien-Jung; Jeng, Sheng-Long; Huang, Rong-Tan; Tsay, Leu-Wen

2017-01-01

The increased thermal efficiency of fossil power plants calls for the development of advanced creep-resistant alloy steels like T92. In this study, microstructures found in the heat-affected zone (HAZ) of a T92 steel weld were simulated to evaluate their creep-rupture-life at elevated temperatures. An infrared heating system was used to heat the samples to 860 °C (around AC1), 900 °C (slightly below AC3), and 940 °C (moderately above AC3) for one minute, before cooling to room temperature. The simulated specimens were then subjected to a conventional post-weld heat treatment (PWHT) at 750 °C for two hours, where both the 900 °C and 940 °C simulated specimens had fine grain sizes. In the as-treated condition, the 900 °C simulated specimen consisted of fine lath martensite, ferrite subgrains, and undissolved carbides, while residual carbides and fresh martensite were found in the 940 °C simulated specimen. The results of short-term creep tests indicated that the creep resistance of the 900 °C and 940 °C simulated specimens was poorer than that of the 860 °C simulated specimens and the base metal. Moreover, simulated T92 steel samples had higher creep strength than the T91 counterpart specimens. PMID:28772500

Effect of Cold Storage on Shear-induced Platelet Aggregation and Clot Strength

DTIC Science & Technology

2014-09-01

stored as 1-mL aliquots in 15-mL polypropylene centri- fuge tubes with gentle agitation either at RT (20Y24-C) or at 4-C (1Y6-C). Single AP units...vivo aging at 37-C. Brit J Haematol. 1995;91:212 218. 16. Babic AM, Josefsson EC, Bergmeier W, Wagner DD, Kaufman RM, Silberstein LE , et al. In vitro

To cool, but not too cool: that is the question--immersion cooling for hyperthermia.

PubMed

Taylor, Nigel A S; Caldwell, Joanne N; Van den Heuvel, Anne M J; Patterson, Mark J

2008-11-01

Patient cooling time can impact upon the prognosis of heat illness. Although ice-cold-water immersion will rapidly extract heat, access to ice or cold water may be limited in hot climates. Indeed, some have concerns regarding the sudden cold-water immersion of hyperthermic individuals, whereas others believe that cutaneous vasoconstriction may reduce convective heat transfer from the core. It was hypothesized that warmer immersion temperatures, which induce less powerful vasoconstriction, may still facilitate rapid cooling in hyperthermic individuals. Eight males participated in three trials and were heated to an esophageal temperature of 39.5 degrees C by exercising in the heat (36 degrees C, 50% relative humidity) while wearing a water-perfusion garment (40 degrees C). Subjects were cooled using each of the following methods: air (20-22 degrees C), cold-water immersion (14 degrees C), and temperate-water immersion (26 degrees C). The time to reach an esophageal temperature of 37.5 degrees C averaged 22.81 min (air), 2.16 min (cold), and 2.91 min (temperate). Whereas each of the between-trial comparisons was statistically significant (P < 0.05), cooling in temperate water took only marginally longer than that in cold water, and one cannot imagine that the 45-s cooling time difference would have any meaningful physiological or clinical implications. It is assumed that this rapid heat loss was due to a less powerful peripheral vasoconstrictor response, with central heat being more rapidly transported to the skin surface for dissipation. Although the core-to-water thermal gradient was much smaller with temperate-water cooling, greater skin and deeper tissue blood flows would support a superior convective heat delivery. Thus, a sustained physiological mechanism (blood flow) appears to have countered a less powerful thermal gradient, resulting in clinically insignificant differences in heat extraction between the cold and temperate cooling trials.

Population variation in isotopic composition of shorebird feathers: Implications for determining molting grounds

USGS Publications Warehouse

Torres-Dowdall, J.; Farmer, A.H.; Bucher, E.H.; Rye, R.O.; Landis, G.

2009-01-01

Stable isotope analyses have revolutionized the study of migratory connectivity. However, as with all tools, their limitations must be understood in order to derive the maximum benefit of a particular application. The goal of this study was to evaluate the efficacy of stable isotopes of C, N, H, O and S for assigning known-origin feathers to the molting sites of migrant shorebird species wintering and breeding in Argentina. Specific objectives were to: 1) compare the efficacy of the technique for studying shorebird species with different migration patterns, life histories and habitat-use patterns; 2) evaluate the grouping of species with similar migration and habitat use patterns in a single analysis to potentially improve prediction accuracy; and 3) evaluate the potential gains in prediction accuracy that might be achieved from using multiple stable isotopes. The efficacy of stable isotope ratios to determine origin was found to vary with species. While one species (White-rumped Sandpiper, Calidris fuscicollis) had high levels of accuracy assigning samples to known origin (91% of samples correctly assigned), another (Collared Plover, Charadrius collaris) showed low levels of accuracy (52% of samples correctly assigned). Intra-individual variability may account for this difference in efficacy. The prediction model for three species with similar migration and habitat-use patterns performed poorly compared with the model for just one of the species (71% versus 91% of samples correctly assigned). Thus, combining multiple sympatric species may not improve model prediction accuracy. Increasing the number of stable isotopes in the analyses increased the accuracy of assigning shorebirds to their molting origin, but the best combination - involving a subset of all the isotopes analyzed - varied among species.

Large-scale meta-analysis of interleukin-1 beta and interleukin-1 receptor antagonist polymorphisms on risk of radiographic hip and knee osteoarthritis and severity of knee osteoarthritis.

PubMed

Kerkhof, H J M; Doherty, M; Arden, N K; Abramson, S B; Attur, M; Bos, S D; Cooper, C; Dennison, E M; Doherty, S A; Evangelou, E; Hart, D J; Hofman, A; Javaid, K; Kerna, I; Kisand, K; Kloppenburg, M; Krasnokutsky, S; Maciewicz, R A; Meulenbelt, I; Muir, K R; Rivadeneira, F; Samuels, J; Sezgin, M; Slagboom, E; Smith, A J P; Spector, T D; Tamm, A; Tamm, A; Uitterlinden, A G; Wheeler, M; Zhai, G; Zhang, W; van Meurs, J B J; Valdes, A M

2011-03-01

To clarify the role of common genetic variation in the Interleukin-1β (IL1B) and Interleukin-1R antagonist (IL1RN) genes on risk of knee and hip osteoarthritis (OA) and severity of knee OA by means of large-scale meta-analyses. We searched PubMed for articles assessing the role of IL1B and IL1RN polymorphisms/haplotypes on the risk of hip and/or knee OA. Novel data were included from eight unpublished studies. Meta-analyses were performed using fixed- and random-effects models with a total of 3595 hip OA and 5013 knee OA cases, and 6559 and 9132 controls respectively. The role of ILRN haplotypes on radiographic severity of knee OA was tested in 1918 cases with Kellgren-Lawrence (K/L) 1 or 2 compared to 199 cases with K/L 3 or 4. The meta-analysis of six published studies retrieved from the literature search and eight unpublished studies showed no evidence of association between common genetic variation in the IL1B or IL1RN genes and risk of hip OA or knee OA (P>0.05 for rs16944, rs1143634, rs419598 and haplotype C-G-C (rs1143634, rs16944 and rs419598) previously implicated in risk of hip OA). The C-T-A haplotype formed by rs419598, rs315952 and rs9005, previously implicated in radiographic severity of knee OA, was associated with reduced severity of knee OA (odds ratio (OR)=0.71 95%CI 0.56-0.91; P=0.006, I(2)=74%), and achieved borderline statistical significance in a random-effects model (OR=0.61 95%CI 0.35-1.06 P=0.08). Common genetic variation in the Interleukin-1 region is not associated with prevalence of hip or knee OA but our data suggest that IL1RN might have a role in severity of knee OA. Copyright © 2010 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.

23 CFR Appendix C to Subpart A of... - Federal-Aid Highway Contractors Annual EEO Report (Form PR-1391)

Code of Federal Regulations, 2013 CFR

2013-04-01

... 23 Highways 1 2013-04-01 2013-04-01 false Federal-Aid Highway Contractors Annual EEO Report (Form PR-1391) C Appendix C to Subpart A of Part 230 Highways FEDERAL HIGHWAY ADMINISTRATION, DEPARTMENT OF... Part 230—Federal-Aid Highway Contractors Annual EEO Report (Form PR-1391) EC14OC91.000 ...

Association studies on the bovine lipoprotein lipase gene polymorphism with growth and carcass quality traits in Qinchuan cattle.

PubMed

Gui, Linsheng; Jia, Cuiling; Zhang, Yaran; Zhao, Chunping; Zan, Linsen

2016-04-01

Lipoprotein lipase (LPL) is considered as an essential enzyme in lipid deposition and tissue metabolism. It has been proposed to be a lead candidate gene for genetic markers of lipid deposition and energy balance. In this paper, polymorphisms in the LPL gene were investigated in 554 Chinese Qinchuan cattle by PCR-RFLP and DNA sequencing. Seven single nucleotide polymorphisms (SNPs) were identified, which included one mutation (g.91C > T) in the 5'untranslated region (UTR), four synonymous mutations (g.17015A > G, g.18362G > A, g.18377T > C and g.19873T > C) and two mutations (g.25225A > G and g.25316T > G) in the 3'UTR. The frequencies of SNP g.18377T > C and g.25316T > G were skewed from Hardy-Weinberg equilibrium in all the samples (chi-square test, P < 0.05). An association analysis showed that five loci (except for g.91C > T and g.18377T > C) were significantly correlated with some growth and carcass quality traits. These results demonstrate that LPL might be a potential candidate gene for marker-assisted selection (MAS). Copyright © 2016. Published by Elsevier Ltd.

Effect of microstructure on the zinc phosphate conversion coatings on magnesium alloy AZ91

NASA Astrophysics Data System (ADS)

Van Phuong, Nguyen; Moon, Sungmo; Chang, Doyon; Lee, Kyu Hwan

2013-01-01

The effect of the microstructure, particularly of β-Mg17Al12 phase, on the formation and growth of zinc phosphate conversion coatings on magnesium alloy AZ91 (AZ91) was studied. The zinc phosphate coatings were formed on AZ91 with different microstructures produced by heat treatment. The effect of the microstructure on the zinc phosphate coatings were examined using optical microscope (OM), X-ray diffraction (XRD), coatings weight and etching weight balances, scanning electron microscopy (SEM) and salt immersion test. Results showed that as-cast AZ91 contained a high volume fraction of the β-Mg17Al12 phase and it was dissolved into α-Mg phase during heat treatment at 400 °C. The β-phase became center for hydrogen evolution during phosphating reaction (cathodic sites). The decreased volume fraction of the β-phase caused decreasing both coatings weight and etching weight of the phosphating process. However, it increased the crystal size of the coatings and improved corrosion resistance of AZ91 by immersing in 0.5 M NaCl solution. Results also showed that the structure of the zinc phosphate conversion on AZ91 consisted of two layers: an outer crystal Zn3(PO4)2·4H2O (hopeite) and an inner which was mainly composed of MgZn2(PO4)2 and Mg3(PO4)2. A mechanism for the formation of two layers of the coatings was also proposed in this study.

Heat Transfer Measurements of Internally Finned Rotating Heat Pipes.

DTIC Science & Technology

1983-12-01

42 C. RESULTS OF STRAIGHT 22 FIN CONDENSER o . o . 51 D. RESULTS OF HELICAL 14 AND 16 FIN CONDENSER o 51 E. RESULTS OF HELICAL 36 FIN...88 C.6 RESULTS OF STRAIGHT 22 FIN CONDENSER AT 2800 RPM . . . . . . . . . . . . . . . . . . . . 89 C.7 RESULTS OF HELICAL 16 FIN CONDENSER ...AT 700 RPM . 90 C.8 RESULTS OF HELICAL 16 PIN CONDENSER AT 1600 RPM . 91 C.9 RESULTS OF HELICAL 16 PIN CONDENSER AT 2800 RPM . 92 C. 10 RESULTS OF

40 CFR 91.312 - Analytical gases.

Code of Federal Regulations, 2012 CFR

2012-07-01

... nitrogen as the predominant diluent with the balance oxygen. (e) Fuel for the hydrocarbon flame ionization... operation. (1) Purified nitrogen, also referred to as “zero-grade nitrogen” (Contamination≤1 ppm C, ≤1 ppm... mixture (40±2 percent hydrogen, balance helium) (Contamination≤1 ppm C, ≤400 ppm CO) (4) Purified...

40 CFR 91.312 - Analytical gases.

Code of Federal Regulations, 2011 CFR

2011-07-01

... nitrogen as the predominant diluent with the balance oxygen. (e) Fuel for the hydrocarbon flame ionization... operation. (1) Purified nitrogen, also referred to as “zero-grade nitrogen” (Contamination≤1 ppm C, ≤1 ppm... mixture (40±2 percent hydrogen, balance helium) (Contamination≤1 ppm C, ≤400 ppm CO) (4) Purified...

40 CFR 91.312 - Analytical gases.

Code of Federal Regulations, 2010 CFR

2010-07-01

... nitrogen as the predominant diluent with the balance oxygen. (e) Fuel for the hydrocarbon flame ionization... operation. (1) Purified nitrogen, also referred to as “zero-grade nitrogen” (Contamination≤1 ppm C, ≤1 ppm... mixture (40±2 percent hydrogen, balance helium) (Contamination≤1 ppm C, ≤400 ppm CO) (4) Purified...

40 CFR 91.312 - Analytical gases.

Code of Federal Regulations, 2013 CFR

2013-07-01

... nitrogen as the predominant diluent with the balance oxygen. (e) Fuel for the hydrocarbon flame ionization... operation. (1) Purified nitrogen, also referred to as “zero-grade nitrogen” (Contamination≤1 ppm C, ≤1 ppm... mixture (40±2 percent hydrogen, balance helium) (Contamination≤1 ppm C, ≤400 ppm CO) (4) Purified...

Pantoea applied genomics to understand and improve biocontrol activity against fire blight

USDA-ARS?s Scientific Manuscript database

Pantoea agglomerans and P. vagans (ex. Erwinia herbicola) are common epiphytes of pome fruit flowers and three strains (E325, P10c, C9-1) have been commercially developed as effective biocontrol products for managing fire blight (Erwinia amylovora). Antibiotics as a standard, reliable chemical optio...

32 CFR 2001.91 - Other agency reporting requirements.

Code of Federal Regulations, 2010 CFR

2010-07-01

... with section 1.7(c)(3) of the Order and § 2001.13(b). (c) Fundamental classification guidance review. The initial fundamental guidance review is to be completed no later than June 27, 2012. Agency heads shall provide a detailed report summarizing the results of each classification guidance review to ISOO...

Joint Demodulation of Low-Entropy Narrowband Cochannel Signals

DTIC Science & Technology

2006-12-01

Linear prediction: A tutorial review,” IEEE Proceedings, vol. 63, pp. 561–580, April 1975. [91] R. G. Brown and P. Y. C. Hwang , Introduction to Random...48 B. SECOND ORDER PREDICTOR . . . . . . . . . . . . . . . . . 49 C. KALMAN FILTER...38 4.1 Prediction algorithm based on the Kalman filter . . . . . . . . . . . . . . . . 52 4.2 self

78 FR 21494 - Union Pacific Railroad Company-Abandonment Exemption-in Washington County, Idaho

Federal Register 2010, 2011, 2012, 2013, 2014

2013-04-10

... Pacific Railroad Company--Abandonment Exemption--in Washington County, Idaho Union Pacific Railroad... at Weiser, in Washington County, Idaho. The line traverses United States Postal Service Zip Code... Bingham & Bonneville Counties, Idaho, 360 I.C.C. 91 (1979). To address whether this condition adequately...

76 FR 16856 - Proposed Collection; Comment Request for Regulation Project

Federal Register 2010, 2011, 2012, 2013, 2014

2011-03-25

... Reduction Act of 1995, Public Law 104-13 (44 U.S.C. 3506(c)(2)(A)). Currently, the IRS is soliciting comments concerning existing final regulations, REG-208156-91 (TD 8929), Accounting for Long-Term Contracts... the Internet at [email protected] . SUPPLEMENTARY INFORMATION: Title: Accounting for Long-Term...

Evaluation of BAUER High Pressure Breathing Air P-2 Purification System

DTIC Science & Technology

1991-08-01

and is a coalescing type separator that removes oil and water vapors suspended in the compressed air . The molecular sieve is made to adsorb oil and...filtering, moisture separation, and prevents compressed air return from the charged air storage flasks to the compressor during unit shutdown. A manual...1111111111111 1111 IE IH fil91i C NAVY EXPERIMENTAL DIVING UNIT REPORT NO. 10-91 EVALUATION OF BAUER HIGH PRESSURE BREATHING AIR P-2 PURIFICATION SYSTEM GEORGE D

WWII (World War II) Era Building Demolition and Renovation Cost Estimator (ESTER) 1.0 User’s Manual

DTIC Science & Technology

1988-07-01

S.F., fiberglass, R-13 */ Idefine BLO F R19 0.91 /* S.F., fiberglass, R-19 */ Idefine B M RII 0.54 /* S.F., mineral wool , R-1l 1i /define W M RI13...0.81 /* S.F., mineral wool , R-13 */ idefine BLOMR19 1.16 /* S.F., mineral wool , R-19 */ .define BSW C R26 0.91 /* S.F., cellulose, R-26 */ Idefine BL

Emergency Preparedness Plan. DeWitt Army Community Hospital, Fort Belvoir, Virginia,

DTIC Science & Technology

1997-12-31

demand, dispatch at least one Respiratory Specialist (91V) to the ER rn Department of Familv Practice or senior representative will: (1) /Wnni™ and...be prepared for are: a. Two (2) messengers/runners to the EOC and Triage area(s). b. One or more Respiratory Specialist (91V) to the ER. c. A...Depressors 1 bx Shave Prep Set 50 ea Skin Closure, Adhesive 2 bx Stethoscope 5 ea Dental Mirrors 2 ea Nasal Speculum 2 ea Betadine Surgi-prep Sponge

Identification and Characterization of Intercritical Heat-Affected Zone in As-Welded Grade 91 Weldment

NASA Astrophysics Data System (ADS)

Wang, Yiyu; Kannan, Rangasayee; Li, Leijun

2016-12-01

A metallurgical method is proposed for locating the intercritical heat-affected zone in the as-welded Grade 91 steel. New austenitic grains, preferentially formed along the original prior austenite grain boundaries, are characterized to contain finer M23C6 carbides and higher strain levels than the original prior austenite grains. Kurdjumov-Sachs Group 1 variant pairs, with a low misorientation of 7 deg within a martensitic block, are identified as the dominant variants in the new PAGs.

Environmental Assessment: Proposed Consolidated Warehouse, Hill Air Force Base, Utah

DTIC Science & Technology

2011-02-01

can be evaluated. It...420 916 180 243 50 3 37 206 50 7 G 43 91 5 508 57 0 47 0 847 896 341 50 5 5 39 50 9 24 0 58 1 5 16 42 849 13 77 883 C 295 840 430 125 U -01-HL- 0164m U ...91-WC-687m U -95-WC-28OP U -02-HL-0008m Proposed Action (Alternative A) Future Training Complex Alternate Location (Alternative D) Fu tu re Au to m

The Design and Implementation of the Ariel Active Database Rule System

DTIC Science & Technology

1991-10-01

but only as a main-memory prototype. The POSTGRES rule system (PRS) [SHP88, SRH90] and the Starburst rule system (SRS) [WCL91, HCL+90] have been...query language of POSTGRES for specifying data definition commands, queries and updates [SRH90]. POSTQUEL commands retrieve, append, delete, and replace...placed on an arbitrary attribute (e.g., one without an index) ( POSTGRES rule system [SHP88, SHP89, SR1I90], HiPAC [C+891, DIPS [SLR89], Alert [SPAM91

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ballinger, Ronald G

A material that resists lead-bismuth eutectic (LBE) attack and retains its strength at 700°C would be an enabling technology for LBE-cooled reactors. No single alloy currently exists that can economically meet the required performance criteria of high strength and corrosion resistance. A Functionally Graded Composite (FGC) was developed with layers engineered to perform these functions. F91 was chosen as the structural layer of the composite for its strength and radiation resistance. Fe-12Cr-2Si, an alloy developed from previous work in the Fe-Cr-Si system, was chosen as the corrosion-resistant cladding layer because of its chemical similarity to F91 and its superior corrosionmore » resistance in both oxidizing and reducing environments. Fe-12Cr-2Si experienced minimal corrosion due to its self-passivation in oxidizing and reducing environments. Extrapolated corrosion rates are below one micron per year at 700ï°C. Corrosion of F91 was faster, but predictable and manageable. Diffusion studies showed that 17 microns of the cladding layer will be diffusionally diluted during the three year life of fuel cladding. 33 microns must be accounted for during the sixty year life of coolant piping. 5 cm coolant piping and 6.35 mm fuel cladding preforms were produced on a commercial scale by weld-overlaying Fe-12Cr-2Si onto F91 billets and co-extruding them. An ASME certified weld was performed followed by the prescribed quench-and-tempering heat treatment for F91. A minimal heat affected zone was observed, demonstrating field weldability. Finally, corrosion tests were performed on the fabricated FGC at 700ï°C after completely breaching the cladding in a small area to induce galvanic corrosion at the interface. None was observed. This FGC has significant impacts on LBE reactor design. The increases in outlet temperature and coolant velocity allow a large increase in power density, leading to either a smaller core for the same power rating or more power output for the same size core. This FGC represents an enabling technology for LBE cooled fast reactors.« less

Differentiation-dependent expression of hypothetical proteins in the neuroblastoma cell line N1E-115.

PubMed

Oh, Ji-eun; Karlmark, Karlin Raja; Shin, Jooho; Hengstschläger, Markus; Lubec, Gert

2006-05-15

Several protein cascades, including signaling, cytoskeletal, chaperones, metabolic, and antioxidant proteins, have been shown to be involved in the process of neuronal differentiation (ND) of neuroblastoma cell lines. No systematic approach to detect hitherto unknown and unnamed proteins or structures that have been predicted upon nucleic acid sequences in ND has been published so far. We therefore decided to screen hypothetical protein (HP) expression by protein profiling. Two-dimensional gel electrophoresis with subsequent matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/TOF) identification was used for expression analysis of undifferentiated and dimethylsulfoxide-induced neuronally differentiated N1E-115 cells. We unambiguously identified six HPs: Q8C520, Q99LF4, Q9CXS1, Q9DAF8, Q91WT0, and Q8C5G2. A prefoldin domain in Q91WT0, a t-SNARE domain in Q9CXS1, and a bromodomain were observed in Q8C5G2. For the three remaining proteins, no putative function using Pfam, BLOCKS, PROSITE, PRINTS, InterPro, Superfamily, CoPS, and ExPASy could be assigned. While two proteins were present in both cell lines, Q9CXS1 was switched off (i.e., undetectably low) in differentiated cells only, and Q9DAF8, Q91WT0, and Q8C5G2 were switched on in differentiated cells exclusively. Herein, using a proteomic approach suitable for screening and identification of HP, we present HP structures that have been only predicted so far based upon nucleic acid sequences. The four differentially regulated HPs may play a putative role in the process of ND. (c) 2006 Wiley-Liss, Inc.

Novel Mechanism for Buffering Dietary Salt in Humans: Effects of Salt Loading on Skin Sodium, Vascular Endothelial Growth Factor C, and Blood Pressure.

PubMed

Selvarajah, Viknesh; Mäki-Petäjä, Kaisa M; Pedro, Liliana; Bruggraber, Sylvaine F A; Burling, Keith; Goodhart, Anna K; Brown, Morris J; McEniery, Carmel M; Wilkinson, Ian B

2017-11-01

High dietary sodium intake triggers increased blood pressure (BP). Animal studies show that dietary salt loading results in dermal Na + accumulation and lymphangiogenesis mediated by VEGF-C (vascular endothelial growth factor C), both attenuating the rise in BP. Our objective was to determine whether these mechanisms function in humans. We assessed skin electrolytes, BP, and plasma VEGF-C in 48 healthy participants randomized to placebo (70 mmol sodium/d) and slow sodium (200 mmol/d) for 7 days. Skin Na + and K + concentrations were measured in mg/g of wet tissue and expressed as the ratio Na + :K + to correct for variability in sample hydration. Skin Na + :K + increased between placebo and slow sodium phases (2.91±0.08 versus 3.12±0.09; P =0.01). In post hoc analysis, there was a suggestion of a sex-specific effect, with a significant increase in skin Na + :K + in men (2.59±0.09 versus 2.88±0.12; P =0.008) but not women (3.23±0.10 versus 3.36±0.12; P =0.31). Women showed a significant increase in 24-hour mean BP with salt loading (93±1 versus 91±1 mm Hg; P <0.001) while men did not (96±2 versus 96±2 mm Hg; P =0.91). Skin Na + :K + correlated with BP, stroke volume, and peripheral vascular resistance in men but not in women. No change was noted in plasma VEGF-C. These findings suggest that the skin may buffer dietary Na + , reducing the hemodynamic consequences of increased salt, and this may be influenced by sex. © 2017 The Authors.

CYP2C9 and CYP2C19 genetic polymorphisms: frequencies in the south Indian population.

PubMed

Jose, Rosemary; Chandrasekaran, Adithan; Sam, Soya Sisy; Gerard, Nathalie; Chanolean, Shashindran; Abraham, Benny K; Satyanarayanamoorthy, K; Peter, Anitha; Rajagopal, Krishnamoorthy

2005-02-01

The aim of the study was to establish the frequencies of CYP2C9*1, *2, *3 and CYP2C19*1, *2 and *3 in the south Indian population and to compare them with the inter-racial distribution of the CYP2C9 and CYP2C19 genetic polymorphisms. Genotyping analyses of CYP2C9 and CYP2C19 were conducted in unrelated, healthy volunteers from the three south Indian states of Andhra Pradesh, Karnataka and Kerala, by the polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP). The allele frequencies of the populations of these three states were then pooled with our previous genotyping data of Tamilians (also in south India), to arrive at the distribution of CYP2C9 and CYP2C19 alleles in the south Indian population. Frequencies of CYP2C9 and CYP2C19 alleles and genotypes among various populations were compared using the two-tailed Fisher's exact test. The frequencies of CYP2C9*1, *2 and *3 in the south Indian population were 0.88 (95% CI 0.85-0.91), 0.04 (95% CI 0.02-0.06) and 0.08 (95% CI 0.06-0.11), respectively. The frequencies of CYP2C9 genotypes *1/*1, *1/*2, *1/*3, *2/*2, *2/*3 and *3/*3 were 0.78 (95% CI 0.74-0.82), 0.05 (95% CI 0.03-0.07), 0.15 (95% CI 0.12-0.18), 0.01 (95% CI 0.0-0.02), 0.01 (95% CI 0.0-0.02) and 0.0, respectively. CYP2C19*1, *2 and *3 frequencies were 0.64 (95% CI 0.60-0.68), 0.35 (95% CI 0.31-0.39) and 0.01 (95% CI 0.0-0.03), respectively. As a result of a significant heterogeneity, the data on CYP2C19 genotype frequencies were not pooled. The frequency of CYP2C9*2 mutant alleles in south Indians was higher than in Chinese and Caucasians, while CYP2C9*3 was similar to Caucasians. CYP2C19*2 was higher than in other major populations reported so far. The relatively high CYP2C19 poor-metabolizer genotype frequency of 12.6% indicates that over 28 million south Indians are poor metabolizers of CYP2C19 substrates.

Racial Disparities in HIV Care Extend to Common Comorbidities: Implications for Implementation of Interventions to Reduce Disparities in HIV Care.

PubMed

Richardson, Kelly K; Bokhour, Barbara; McInnes, D Keith; Yakovchenko, Vera; Okwara, Leonore; Midboe, Amanda M; Skolnik, Avy; Vaughan-Sarrazin, Mary; Asch, Steven M; Gifford, Allen L; Ohl, Michael E

2016-01-01

Prior studies have described racial disparities in the quality of care for persons with HIV infection, but it is unknown if these disparities extend to common comorbid conditions. To inform implementation of interventions to reduce disparities in HIV care, we examined racial variation in a set of quality measures for common comorbid conditions among Veterans in care for HIV in the United States. The cohort included 23,974 Veterans in care for HIV in 2013 (53.4% black; 46.6% white). Measures extracted from electronic health record and administrative data were receipt of combination antiretroviral therapy (cART), HIV viral control (serum RNA < 200 copies/ml among those on cART), hypertension control (blood pressure < 140/90 mm Hg among those with hypertension), diabetes control (hemoglobin A1C < 9% among those with diabetes), lipid monitoring, guideline-concordant antidepressant prescribing, and initiation and engagement in substance use disorder (SUD) treatment. Black persons were less likely than their white counterparts to receive cART (90.2% vs. 93.2%, p<.001), and experience viral control (84.6% vs. 91.3%, p<.001), hypertension control (61.9% vs. 68.3%, p<.001), diabetes control (85.5% vs. 89.5%, p<.001), and lipid monitoring (81.5% vs. 85.2%, p<.001). Initiation and engagement in SUD treatment were similar among blacks and whites. Differences remained after adjusting for age, comorbidity, retention in HIV care, and a measure of neighborhood social disadvantage created from census data. Implementation of interventions to reduce racial disparities in HIV care should comprehensively address and monitor processes and outcomes of care for key comorbidities. Published by Elsevier Inc.

Implementation and evaluation of a pharmacist-led hypertension management service in primary care: outcomes and methodological challenges.

PubMed

Bajorek, Beata; Lemay, Kate S; Magin, Parker; Roberts, Christopher; Krass, Ines; Armour, Carol L

2016-01-01

Suboptimal utilisation of pharmacotherapy, non-adherence to prescribed treatment, and a lack of monitoring all contribute to poor blood (BP) pressure control in patients with hypertension. The objective of this study was to evaluate the implementation of a pharmacist-led hypertension management service in terms of processes, outcomes, and methodological challenges. A prospective, controlled study was undertaken within the Australian primary care setting. Community pharmacists were recruited to one of three study groups: Group A (Control - usual care), Group B (Intervention), or Group C (Short Intervention). Pharmacists in Groups B and C delivered a service comprising screening and monitoring of BP, as well as addressing poor BP control through therapeutic adjustment and adherence strategies. Pharmacists in Group C delivered the shortened version of the service. Significant changes to key outcome measures were observed in Group C: reduction in systolic and diastolic BPs at the 3-month visit (P<0.01 and P<0.01, respectively), improvement in medication adherence scores (P=0.01), and a slight improvement in quality of life (EQ-5D-3L Index) scores (P=0.91). There were no significant changes in Group B (the full intervention), and no differences in comparison to Group A (usual care). Pharmacists fed-back that patient recruitment was a key barrier to service implementation, highlighting the methodological implications of screening. A collaborative, pharmacist-led hypertension management service can help monitor BP, improve medication adherence, and optimise therapy in a step-wise approach. However, blood pressure screening can effect behaviour change in patients, presenting methodological challenges in the evaluation of services in this context.

Implementation and evaluation of a pharmacist-led hypertension management service in primary care: outcomes and methodological challenges

PubMed Central

2016-01-01

Background: Suboptimal utilisation of pharmacotherapy, non-adherence to prescribed treatment, and a lack of monitoring all contribute to poor blood (BP) pressure control in patients with hypertension. Objective: The objective of this study was to evaluate the implementation of a pharmacist-led hypertension management service in terms of processes, outcomes, and methodological challenges. Method: A prospective, controlled study was undertaken within the Australian primary care setting. Community pharmacists were recruited to one of three study groups: Group A (Control – usual care), Group B (Intervention), or Group C (Short Intervention). Pharmacists in Groups B and C delivered a service comprising screening and monitoring of BP, as well as addressing poor BP control through therapeutic adjustment and adherence strategies. Pharmacists in Group C delivered the shortened version of the service. Results: Significant changes to key outcome measures were observed in Group C: reduction in systolic and diastolic BPs at the 3-month visit (P<0.01 and P<0.01, respectively), improvement in medication adherence scores (P=0.01), and a slight improvement in quality of life (EQ-5D-3L Index) scores (P=0.91). There were no significant changes in Group B (the full intervention), and no differences in comparison to Group A (usual care). Pharmacists fed-back that patient recruitment was a key barrier to service implementation, highlighting the methodological implications of screening. Conclusion: A collaborative, pharmacist-led hypertension management service can help monitor BP, improve medication adherence, and optimise therapy in a step-wise approach. However, blood pressure screening can effect behaviour change in patients, presenting methodological challenges in the evaluation of services in this context. PMID:27382427

Mycobacterium tuberculosis NmtR harbors a nickel sensing site with parallels to Escherichia coli RcnR†

PubMed Central

Reyes-Caballero, Hermes; Lee, Chul Won; Giedroc, David P.

2011-01-01

Mycobacterium tuberculosis NmtR is a Ni(II)/Co(II)-sensing metalloregulatory protein from the extensively studied ArsR/SmtB family. Two Ni(II) ions bind to the NmtR dimer to form octahedral coordination complexes with stepwise binding affinities of KNi1=1.2 (±0.1) × 1010 and KNi2=0.7 (±0.4) × 1010 M-1 (pH 7.0). A glutamine scanning mutagenesis approach reveals that Asp91, His93, His104 and His107, all contained within the C-terminal α5 helix, and His3 as part of the conserved α-NH2-Gly2-His3-Gly4 motif at the N-terminus make significant contributions to the magnitude of KNi. In contrast, substitution of residues from the C-terminal region, His109, Asp114 and His116, previously implicated in Ni(II) binding and metalloregulation in cells, gives rise to wild-type KNi and Ni(II)-dependent allosteric coupling free energies. Interestingly, deletion of residues 112-120 in the C-terminal region (Δ111 NmtR) reduces the Ni(II) binding stoichiometry to one per dimer and greatly reduces Ni(II) responsiveness. H3Q and Δ111 NmtRs also show clear perturbations in the rank order of metal responsiveness to Ni(II), Co(II) and Zn(II) that is distinct from wild-type NmtR. 15N relaxation experiments with apo-NmtR reveal that both N-terminal (residues 2-14) and C-terminal (residues 110-120) regions are unstructured in solution, and this property likely dictates the metal specificity profile characteristic of the Ni(II)-sensor NmtR relative to other ArsR family regulators. PMID:21819125

NASA Astrophysics Data System (ADS)

Scapin, M.; Peroni, L.; Fichera, C.; Cambriani, A.

2014-08-01

High chromium ferritic/martensitic steel T91 (9% Cr, 1% Mo), on account of its radiation resistance, is a candidate material for nuclear reactor applications. Its joining by an impact method to create a cold joint is tested in the realm of scoping tests toward the safe operation of nuclear fuels, encapsulated in representative T91 materials. Hitherto, T91 mechanical characterization at high strain rates is relatively unknown, particularly, in relation to impact joining and also to nuclear accidents. In this study, the mechanical characterization of T91 steel was performed in tension by varying the strain-rate (10-3 up to 104 s-1) and temperature (20-800°C) on dog-bone specimens, using standard testing machines or Hopkinson Bar apparati. As expected, the material is both temperature and strain-rate sensitive and different sets of parameters for the Johnson-Cook strength model were extracted via a numerical inverse procedure, in order to obtain the most suitable set to be used in this field of applications.

Cofactor composition and function of a H2-sensing regulatory hydrogenase as revealed by Mössbauer and EPR spectroscopy† †Electronic supplementary information (ESI) available: Tables with the simulation parameters and details of the Mössbauer, and EPR spectra (Tables S1–S4). additional EPR and Mössbauer spectra in Fig. S1–S9. See DOI: 10.1039/c5sc01560j Click here for additional data file.

PubMed Central

Roncaroli, Federico; Friedrich, Bärbel; Lenz, Oliver

2015-01-01

The regulatory hydrogenase (RH) from Ralstonia eutropha H16 acts as a sensor for the detection of environmental H2 and regulates gene expression related to hydrogenase-mediated cellular metabolism. In marked contrast to prototypical energy-converting [NiFe] hydrogenases, the RH is apparently insensitive to inhibition by O2 and CO. While the physiological function of regulatory hydrogenases is well established, little is known about the redox cycling of the [NiFe] center and the nature of the iron–sulfur (FeS) clusters acting as electron relay. The absence of any FeS cluster signals in EPR had been attributed to their particular nature, whereas the observation of essentially only two active site redox states, namely Ni-SI and Ni-C, invoked a different operant mechanism. In the present work, we employ a combination of Mössbauer, FTIR and EPR spectroscopic techniques to study the RH, and the results are consistent with the presence of three [4Fe–4S] centers in the small subunit. In the as-isolated, oxidized RH all FeS clusters reside in the EPR-silent 2+ state. Incubation with H2 leads to reduction of two of the [4Fe–4S] clusters, whereas only strongly reducing agents lead to reduction of the third cluster, which is ascribed to be the [4Fe–4S] center in ‘proximal’ position to the [NiFe] center. In the two different active site redox states, the low-spin FeII exhibits distinct Mössbauer features attributed to changes in the electronic and geometric structure of the catalytic center. The results are discussed with regard to the spectral characteristics and physiological function of H2-sensing regulatory hydrogenases. PMID:29142700

Study of microstress state of P91 steel using complementary mechanical Barkhausen, magnetoacoustic emission, and X-ray diffraction techniques

DOE Office of Scientific and Technical Information (OSTI.GOV)

Augustyniak, Bolesław, E-mail: bolek@mif.pg.gda.pl; Piotrowski, Leszek; Maciakowski, Paweł

2014-05-07

The paper deals with assessment of microstress state of martensite P91 steel using three complementary techniques: mechanical Barkhausen emission, magnetoacoustic emission (MAE), and X-ray diffraction (XRD) profile analysis. Magnetic coercivity Hc and microstructure were investigated with inductive magnetometry and magnetic force microscopy (MFM), respectively. Internal stress level of P91 steel was modified by heat treatment. Steel samples were austenitized, quenched, and then tempered at three temperatures (720 °C, 750 °C, and 780 °C) during increasing time (from 15 min up to 240 min). The microstrain level ε{sub i} was evaluated using Williamson–Hall method. It was revealed that during tempering microstrain systematically decreases from ε{sub i} = 2.5more » × 10{sup −3} for as quenched state down to ε{sub i} = 0.3 × 10{sup −3} for well tempered samples. Both mechanical hardness (Vicker's HV) and magnetic hardness (coercivity) decrease almost linearly with decreasing microstrain while the MAE and MBE intensities strongly increase. Tempering leads to evident shift of the MeBN intensity maximum recorded for the first load towards lower applied strain values and to increase of MAE intensity. This indicates that the microstress state deduced by magnetic techniques is correlated with microstrains evaluated with XRD technique.« less

Representation of international authorship across prominent journals in the field of mental retardation.

PubMed

Matson, Michael L; Matso, Johnny L; Lott, Julia D; Logan, James R

2002-01-01

This article is a reflection of international publication trends across prominent journals in the field of mental retardation. Journals reviewed were the Journal of Applied Behavior Analysis, Journal of Intellectual Disability Research, Mental Retardation, and Research in Developmental Disabilities. Percentage of authors from the United States (US) and other countries were evaluated. US authors represented 84-91% in US journals, while US representation in foreign journals was much lower at 21%. The implications of these findings are discussed.

Brief Report: Investigating the Implications of Applying the New DSM-5 Criteria for Diagnosing Autism Spectrum Disorder in a Preschool Population in Singapore

ERIC Educational Resources Information Center

Wong, Chui Mae; Koh, Hwan Cui

2016-01-01

Diagnostic reports for 206 children who underwent an assessment for autism spectrum disorder (ASD) using the DSM-IV-TR criteria, were re-evaluated using the DSM-5 criteria. Mean age of the children at time of diagnosis was 3 years 10 months. Of the 202 children diagnosed with ASD on the DSM-IV-TR, 184 (91.1%) also met the DSM-5 criteria for ASD.…

Dietary Regulation of PTEN Signaling and Mammary Tumor Initiating Cells: Implications for Breast Cancer Prevention

DTIC Science & Technology

2012-07-01

1999 Paradoxical decrease of an adipose-specific protein, adiponectin, in obesity. Biochem Bio- phys Res Commun 257:79–83 10. Barb D, Pazaitou...Y, Pecquery R 2006 Adiponectin mediates antiproliferative and ap- optotic responses in human MCF7 breast cancer cells. Biochem Bio- phys Res Commun ...Noorden S, Wahlstrom T, Coombes RC, Warner M, Gustafsson JA 2002 Estrogen receptor in breast cancer. Endocr Relat Cancer 9:1–13 29. Esslimani-Sahla M

The Third World Arms Market in the 1980’s: Implications for U. S. Policy.

DTIC Science & Technology

1981-06-02

transfer of modern armaments has gone beyond6 the bounds of ordinary laisse - faire economics. This study is best remembered for its early recognition of... styles , and 8 relationships" in arms transfers. As in the Adelphi study, arms transfers are tabulated numerically rather than in dollar terms. Country...or sells the services of DOD personnel such as training or management advice. 19 91. ILI ’a.UK E.B. Rex The Third World Arms Market 92. As written

Impacts of Japanese Colonialism on State and Economic Development in Korea and Taiwan, and Its Implications for Democracy

DTIC Science & Technology

2009-06-01

Ibid., 262–263. 7 John Lonsdale, review of African Perspectives on Colonialism, by A. Adu Boahen, The Journal of African History 91, 3 (1988): 556. 8...81 T . J. Pempel. “Bureaucracy in Japan.” Political Science and Politics, 25, 1(March 1992), 19. 82 Joel Migdal. “The...Bureaucracy and Economic Development in Japan.” Asian Survey, 5, 11, (November 1965), 531. 93 T . J. Pempel and Michio Muramatsu. “The Japanese

List of U.S. Army Research Institute Research and Technical Publications: October 1, 1989, to September 30, 1994. With Author and Subject Index

DTIC Science & Technology

1995-02-01

Control conditions. The implications of these results to military situations were dis- cussed. A pilot field experiment with marching soldiers tested...data for the seven MOS comprising the 96 Career Management Field (CMF). RN 91-42 Requirements-Based Restruc- turing of Army Military Occu- pational...Institute for the Behavioral and Social Sciences (ARI) Fort Gordon Field Unit surveyed and assessed training technology both within and outside of

40 CFR 86.112-91 - Weighing chamber (or room) and microgram balance specifications.

Code of Federal Regulations, 2014 CFR

2014-07-01

... temperature of the chamber in which the particulate filters are conditioned and weighed shall be maintained to within ±10 °F (6 °C) of a set point between 68 °F (20 °C) and 86 °F (30 °C) during all filter conditioning and filter weighing. A continuous recording of the temperature is required. (2) Humidity. The...

40 CFR 86.112-91 - Weighing chamber (or room) and microgram balance specifications.

Code of Federal Regulations, 2012 CFR

2012-07-01

... temperature of the chamber in which the particulate filters are conditioned and weighed shall be maintained to within ±10 °F (6 °C) of a set point between 68 °F (20 °C) and 86 °F (30 °C) during all filter conditioning and filter weighing. A continuous recording of the temperature is required. (2) Humidity. The...