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Sample records for mercuric ion reductase

  1. Biochemical and Structural Properties of a Thermostable Mercuric Ion Reductase from Metallosphaera sedula

    PubMed Central

    Artz, Jacob H.; White, Spencer N.; Zadvornyy, Oleg A.; Fugate, Corey J.; Hicks, Danny; Gauss, George H.; Posewitz, Matthew C.; Boyd, Eric S.; Peters, John W.

    2015-01-01

    Mercuric ion reductase (MerA), a mercury detoxification enzyme, has been tuned by evolution to have high specificity for mercuric ions (Hg2+) and to catalyze their reduction to a more volatile, less toxic elemental form. Here, we present a biochemical and structural characterization of MerA from the thermophilic crenarchaeon Metallosphaera sedula. MerA from M. sedula is a thermostable enzyme, and remains active after extended incubation at 97°C. At 37°C, the NADPH oxidation-linked Hg2+ reduction specific activity was found to be 1.9 μmol/min⋅mg, increasing to 3.1 μmol/min⋅mg at 70°C. M. sedula MerA crystals were obtained and the structure was solved to 1.6 Å, representing the first solved crystal structure of a thermophilic MerA. Comparison of both the crystal structure and amino acid sequence of MerA from M. sedula to mesophillic counterparts provides new insights into the structural determinants that underpin the thermal stability of the enzyme. PMID:26217660

  2. Structure of the detoxification catalyst mercuric ion reductase from Bacillus sp. strain RC607.

    PubMed

    Schiering, N; Kabsch, W; Moore, M J; Distefano, M D; Walsh, C T; Pai, E F

    1991-07-11

    Several hundred million tons of toxic mercurials are dispersed in the biosphere. Microbes can detoxify organo-mercurials and mercury salts through sequential action of two enzymes, organomercury lyase and mercuric ion reductase (MerA). The latter, a homodimer with homology to the FAD-dependent disulphide oxidoreductases, catalyses the reaction NADPH + Hg(II)----NADP+ + H+ + Hg(0), one of the very rare enzymic reactions with metal substrates. Human glutathione reductase serves as a reference molecule for FAD-dependent disulphide reductases and between its primary structure and that of MerA from Tn501 (Pseudomonas), Tn21 (Shigella), p1258 (Staphylococcus) and Bacillus, 25-30% of the residues have been conserved. All MerAs have a C-terminal extension about 15 residues long but have very varied N termini. Although the enzyme from Streptomyces lividans has no addition, from Pseudomonas aeruginosa Tn501 and Bacillus sp. strain RC607 it has one and two copies respectively of a domain of 80-85 residues, highly homologous to MerP, the periplasmic component of proteins encoded by the mer operon. These domains can be proteolytically cleaved off without changing the catalytic efficiency. We report here the crystal structure of MerA from the Gram-positive bacterium Bacillus sp. strain RC607. Analysis of its complexes with nicotinamide dinucleotide substrates and the inhibitor Cd(II) reveals how limited structural changes enable an enzyme to accept as substrate what used to be a dangerous inhibitor. Knowledge of the mode of mercury ligation is a prerequisite for understanding this unique detoxification mechanism.

  3. Structure of the detoxification catalyst mercuric ion reductase from Bacillus sp. strain RC607

    NASA Astrophysics Data System (ADS)

    Schiering, N.; Kabsch, W.; Moore, M. J.; Distefano, M. D.; Walsh, C. T.; Pai, E. F.

    1991-07-01

    SEVERAL hundred million tons of toxic mercurials are dispersed in the biosphere1. Microbes can detoxify organo-mercurials and mercury salts through sequential action of two enzymes, organomercury lyase2 and mercuric ion reductase (MerA) 3-5. The latter, a homodimer with homology to the FAD-dependent disulphide oxidoreductases6, catalyses the reaction NADPH + Hg(II) --> NADP+ + H+Hg(0), one of the very rare enzymic reactions with metal substrates. Human glutathione reductase7,8 serves as a reference molecule for FAD-dependent disulphide reductases and between its primary structure9 and that of MerA from Tn501 (Pseudomonas), Tn21 (Shigella), pI258 (Staphylococcus) and Bacillus, 25-30% of the residues have been conserved10,11. All MerAs have a C-terminal extension about 15 residues long but have very varied N termini. Although the enzyme from Streptomyces lividans has no addition, from Pseudomonas aeruginosa Tn5Ol and Bacillus sp. strain RC607 it has one and two copies respectively of a domain of 80-85 residues, highly homologous to MerP, the periplasmic component of proteins encoded by the mer operon11. These domains can be proteolytically cleaved off without changing the catalytic efficiency3. We report here the crystal structure of MerA from the Gram-positive bacterium Bacillus sp. strain RC607. Analysis of its complexes with nicotinamide dinucleotide substrates and the inhibitor Cd(II) reveals how limited structural changes enable an enzyme to accept as substrate what used to be a dangerous inhibitor. Knowledge of the mode of mercury ligation is a prerequisite for understanding this unique detoxification mechanism.

  4. Structure and dynamics of a compact state of a multidomain protein, the mercuric ion reductase

    SciTech Connect

    Hong, Liang; Sharp, Melissa A.; Poblete, Simon; Biehle, Ralf; Zamponi, Michaela; Szekely, Noemi; Appavou, Marie -Sousai; Winkler, Roland G.; Nauss, Rachel E.; Johs, Alexander; Parks, Jerry M.; Yi, Zheng; Cheng, Xiaolin; Liang, Liyuan; Ohl, Michael; Miller, Susan M.; Richter, Dieter; Gompper, Gerhard; Smith, Jeremy C.

    2014-07-15

    Here, the functional efficacy of colocalized, linked protein domains is dependent on linker flexibility and system compaction. However, the detailed characterization of these properties in aqueous solution presents an enduring challenge. Here, we employ a novel, to our knowledge, combination of complementary techniques, including small-angle neutron scattering, neutron spin-echo spectroscopy, and all-atom molecular dynamics and coarse-grained simulation, to identify and characterize in detail the structure and dynamics of a compact form of mercuric ion reductase (MerA), an enzyme central to bacterial mercury resistance. MerA possesses metallochaperone-like N-terminal domains (NmerA) tethered to its catalytic core domain by linkers. The NmerA domains are found to interact principally through electrostatic interactions with the core, leashed by the linkers so as to subdiffuse on the surface over an area close to the core C-terminal Hg(II)-binding cysteines. How this compact, dynamical arrangement may facilitate delivery of Hg(II) from NmerA to the core domain is discussed.

  5. Structure and Dynamics of a Compact State of a Multidomain Protein, the Mercuric Ion Reductase

    PubMed Central

    Hong, Liang; Sharp, Melissa A.; Poblete, Simón; Biehl, Ralf; Zamponi, Michaela; Szekely, Noemi; Appavou, Marie-Sousai; Winkler, Roland G.; Nauss, Rachel E.; Johs, Alexander; Parks, Jerry M.; Yi, Zheng; Cheng, Xiaolin; Liang, Liyuan; Ohl, Michael; Miller, Susan M.; Richter, Dieter; Gompper, Gerhard; Smith, Jeremy C.

    2014-01-01

    The functional efficacy of colocalized, linked protein domains is dependent on linker flexibility and system compaction. However, the detailed characterization of these properties in aqueous solution presents an enduring challenge. Here, we employ a novel, to our knowledge, combination of complementary techniques, including small-angle neutron scattering, neutron spin-echo spectroscopy, and all-atom molecular dynamics and coarse-grained simulation, to identify and characterize in detail the structure and dynamics of a compact form of mercuric ion reductase (MerA), an enzyme central to bacterial mercury resistance. MerA possesses metallochaperone-like N-terminal domains (NmerA) tethered to its catalytic core domain by linkers. The NmerA domains are found to interact principally through electrostatic interactions with the core, leashed by the linkers so as to subdiffuse on the surface over an area close to the core C-terminal Hg(II)-binding cysteines. How this compact, dynamical arrangement may facilitate delivery of Hg(II) from NmerA to the core domain is discussed. PMID:25028881

  6. Structural characterization of intramolecular Hg2+ transfer between flexibly-linked domains of mercuric ion reductase

    SciTech Connect

    Johs, Alexander; Harwood, Ian M; Parks, Jerry M; Nauss, Rachel; Smith, Jeremy C; Liang, Liyuan; Miller, Susan M

    2011-01-01

    The enzyme mercuric ion reductase, MerA, is the central component of bacterial mercury resistance encoded by the mer operon. Many MerA proteins possess a metallochaperone-like N-terminal domain, NmerA, that can transfer Hg2+ to the catalytic core (Core) for reduction to Hg0. These domains are tethered to the homodimeric Core by ~30-residue linkers that are subject to proteolysis, which has limited structural and functional characterization of the interactions of these domains. Here, we report purification of homogeneous full-length MerA using a fusion protein construct and combine small-angle X-ray and neutron scattering with molecular dynamics simulation to characterize the structure of constructs that mimic the system before and during handoff of Hg2+ from NmerA to the Core. The radii of gyration, distance distribution functions and Kratky plots derived from the small-angle X-ray scattering data are consistent with full-length MerA adopting elongated conformations resulting from flexibility in the linkers to the NmerA domains. The scattering profiles are best reproduced using an ensemble of linker conformations. This flexible attachment of NmerA may facilitate fast and efficient removal of Hg2+ from diverse protein substrates. Using a specific mutant of MerA allowed determination of the position and relative orientation of NmerA to the Core during Hg2+ handoff. The small buried surface area at the site of interaction suggests molecular recognition may be of less importance for the integrity of metal ion transfers between tethered domains than for transfers between separate proteins in metal trafficking pathways.

  7. Structure/Function Analysis of Protein-Protein Interactions and Role of Dynamic Motions in Mercuric Ion Reductase

    SciTech Connect

    Miller, Susan M.

    2005-05-18

    This report summarizes the activities and findings of our structure/function studies of the bacterial detoxification enzyme mercuric ion reductase. The objectives of the work were to obtain crystal structure information for the catalytic core of this enzyme, use the information to investigate the importance of specific parts of the enzyme to its function, and investigate the role of one domain of the enzyme in its function within cells. We describe the accomplishments towards these goals including many structures of the wild type and mutant forms of the enzyme that highlight its interactions with its Hg(II) substrate, elucidation of the role of the N-terminal domain in vitro and in vivo, and elucidation of the roles of at two conserved residues in the core in the mechanism of catalysis.

  8. Mutagenesis of the redox-active disulfide in mercuric ion reductase: Catalysis by mutant enzymes restricted to flavin redox chemistry

    SciTech Connect

    Distefano, M.D.; Au, K.G.; Walsh, C.T. )

    1989-02-07

    Mercuric reductase, a flavoenzyme that possesses a redox-active cystine, Cys{sub 135}Cys{sub 140}, catalyzes the reduction of Hg(II) to Hg(0) by NADPH. As a probe of mechanism, the authors have constructed mutants lacking a redox-active disulfide by eliminating Cys{sub 135} (Ala{sub 135}Cys{sub 140}), Cys{sub 14} (Cys{sub 135}Ala{sub 140}), or both (Ala{sub 135}Ala{sub 140}). Additionally, they have made double mutants that lack Cys{sub 135} (Ala{sub 135}Cys{sub 139}Cys{sub 140}) or Cys{sub 140} (Cys{sub 135}Cys{sub 139}Ala{sub 140}) but introduce a new Cys in place of Gly{sub 139} with the aim of constructing dithiol pairs in the active site that do not form a redox-active disulfide. The resulting mutant enzymes all lack redox-active disulfides and are hence restricted to FAD/FADH{sub 2} redox chemistry. Each mutant enzyme possesses unique physical and spectroscopic properties that reflect subtle differences in the FAD microenvironment. Preliminary evidence for the Ala{sub 135}Cys{sub 139}Cys{sub 14} mutant enzyme suggests that this protein forms a disulfide between the two adjacent Cys residues. Hg(II) titration experiments that correlate the extent of charge-transfer quenching with Hg(II) binding indicate that the Ala{sub 135}Cys{sub 140} protein binds Hg(II) with substantially less avidity than does the wild-type enzyme. All mutant mercuric reductases catalyze transhydrogenation and oxygen reduction reactions through obligatory reduced flavin intermediates at rates comparable to or greater than that of the wild-type enzyme. In multiple-turnover assays which monitored the production of Hg(0), two of the mutant enzymes were observed to proceed through at least 30 turnovers at rates ca. 1000-fold slower than that of wild-type mercuric reductase. They conclude that the Cys{sub 135} and Cys{sub 140} thiols serve as Hg(II) ligands that orient the Hg(II) for subsequent reduction by a reduced flavin intermediate.

  9. A conservative region of the mercuric reductase gene (mera) as a molecular marker of bacterial mercury resistance

    PubMed Central

    Sotero-Martins, Adriana; de Jesus, Michele Silva; Lacerda, Michele; Moreira, Josino Costa; Filgueiras, Ana Luzia Lauria; Barrocas, Paulo Rubens Guimarães

    2008-01-01

    The most common bacterial mercury resistance mechanism is based on the reduction of Hg(II) to Hg0, which is dependent of the mercuric reductase enzyme (MerA) activity. The use of a 431 bp fragment of a conservative region of the mercuric reductase (merA) gene was applied as a molecular marker of this mechanism, allowing the identification of mercury resistant bacterial strains. PMID:24031221

  10. Community Analysis of a Mercury Hot Spring Supports Occurrence of Domain-Specific Forms of Mercuric Reductase

    PubMed Central

    Simbahan, Jessica; Kurth, Elizabeth; Schelert, James; Dillman, Amanda; Moriyama, Etsuko; Jovanovich, Stevan; Blum, Paul

    2005-01-01

    Mercury is a redox-active heavy metal that reacts with active thiols and depletes cellular antioxidants. Active resistance to the mercuric ion is a widely distributed trait among bacteria and results from the action of mercuric reductase (MerA). Protein phylogenetic analysis of MerA in bacteria indicated the occurrence of a second distinctive form of MerA among the archaea, which lacked an N-terminal metal recruitment domain and a C-terminal active tyrosine. To assess the distribution of the forms of MerA in an interacting community comprising members of both prokaryotic domains, studies were conducted at a naturally occurring mercury-rich geothermal environment. Geochemical analyses of Coso Hot Springs indicated that mercury ore (cinnabar) was present at concentrations of parts per thousand. Under high-temperature and acid conditions, cinnabar may be oxidized to the toxic form Hg2+, necessitating mercury resistance in resident prokaryotes. Culture-independent analysis combined with culture-based methods indicated the presence of thermophilic crenarchaeal and gram-positive bacterial taxa. Fluorescence in situ hybridization analysis provided quantitative data for community composition. DNA sequence analysis of archaeal and bacterial merA sequences derived from cultured pool isolates and from community DNA supported the hypothesis that both forms of MerA were present. Competition experiments were performed to assess the role of archaeal merA in biological fitness. An essential role for this protein was evident during growth in a mercury-contaminated environment. Despite environmental selection for mercury resistance and the proximity of community members, MerA retains the two distinct prokaryotic forms and avoids genetic homogenization. PMID:16332880

  11. Community analysis of a mercury hot spring supports occurrence of domain-specific forms of mercuric reductase.

    PubMed

    Simbahan, Jessica; Kurth, Elizabeth; Schelert, James; Dillman, Amanda; Moriyama, Etsuko; Jovanovich, Stevan; Blum, Paul

    2005-12-01

    Mercury is a redox-active heavy metal that reacts with active thiols and depletes cellular antioxidants. Active resistance to the mercuric ion is a widely distributed trait among bacteria and results from the action of mercuric reductase (MerA). Protein phylogenetic analysis of MerA in bacteria indicated the occurrence of a second distinctive form of MerA among the archaea, which lacked an N-terminal metal recruitment domain and a C-terminal active tyrosine. To assess the distribution of the forms of MerA in an interacting community comprising members of both prokaryotic domains, studies were conducted at a naturally occurring mercury-rich geothermal environment. Geochemical analyses of Coso Hot Springs indicated that mercury ore (cinnabar) was present at concentrations of parts per thousand. Under high-temperature and acid conditions, cinnabar may be oxidized to the toxic form Hg2+, necessitating mercury resistance in resident prokaryotes. Culture-independent analysis combined with culture-based methods indicated the presence of thermophilic crenarchaeal and gram-positive bacterial taxa. Fluorescence in situ hybridization analysis provided quantitative data for community composition. DNA sequence analysis of archaeal and bacterial merA sequences derived from cultured pool isolates and from community DNA supported the hypothesis that both forms of MerA were present. Competition experiments were performed to assess the role of archaeal merA in biological fitness. An essential role for this protein was evident during growth in a mercury-contaminated environment. Despite environmental selection for mercury resistance and the proximity of community members, MerA retains the two distinct prokaryotic forms and avoids genetic homogenization.

  12. Placental and Fetal Disposition of Mercuric Ions in Rats Exposed to Methylmercury: Role of Mrp2

    PubMed Central

    Bridges, Christy C.; Joshee, Lucy; Zalups, Rudolfs K.

    2012-01-01

    Methylmercury is a prevalent environmental toxicant that can have deleterious effects on a developing fetus. Previous studies indicate that the multidrug resistance-associated protein 2 (Mrp2) is involved in renal and hepatic export of mercuric ions. Therefore, we hypothesize that Mrp2 is also involved in export of mercuric ions from placental trophoblasts and fetal tissues. To test this hypothesis, we assessed the disposition of mercuric ions in pregnant Wistar and TR– (Mrp2-deficient) rats exposed to a single dose of methylmercury. The amount of mercury in renal tissues (cortex and outer stripe of outer medulla), liver, blood, amniotic fluid, uterus, placentas and fetuses was significantly greater in TR– rats than in Wistar rats. Urinary and fecal elimination of mercury was greater in Wistar dams than in TR– dams. Thus, our findings suggest that Mrp2 may be involved in the export of mercuric ions from maternal and fetal organs following exposure to methylmercury. PMID:23059061

  13. Mercury (II) removal by resistant bacterial isolates and mercuric (II) reductase activity in a new strain of Pseudomonas sp. B50A.

    PubMed

    Giovanella, Patricia; Cabral, Lucélia; Bento, Fátima Menezes; Gianello, Clesio; Camargo, Flávio Anastácio Oliveira

    2016-01-25

    This study aimed to isolate mercury resistant bacteria, determine the minimum inhibitory concentration for Hg, estimate mercury removal by selected isolates, explore the mer genes, and detect and characterize the activity of the enzyme mercuric (II) reductase produced by a new strain of Pseudomonas sp. B50A. The Hg removal capacity of the isolates was determined by incubating the isolates in Luria Bertani broth and the remaining mercury quantified by atomic absorption spectrophotometry. A PCR reaction was carried out to detect the merA gene and the mercury (II) reductase activity was determined in a spectrophotometer at 340 nm. Eight Gram-negative bacterial isolates were resistant to high mercury concentrations and capable of removing mercury, and of these, five were positive for the gene merA. The isolate Pseudomonas sp. B50A removed 86% of the mercury present in the culture medium and was chosen for further analysis of its enzyme activity. Mercuric (II) reductase activity was detected in the crude extract of this strain. This enzyme showed optimal activity at pH 8 and at temperatures between 37 °C and 45 °C. The ions NH4(+), Ba(2+), Sn(2+), Ni(2+) and Cd(2+) neither inhibited nor stimulated the enzyme activity but it decreased in the presence of the ions Ca(2+), Cu(+) and K(+). The isolate and the enzyme detected were effective in reducing Hg(II) to Hg(0), showing the potential to develop bioremediation technologies and processes to clean-up the environment and waste contaminated with mercury.

  14. Structural and functional characterization of mercuric reductase from Lysinibacillus sphaericus strain G1.

    PubMed

    Bafana, Amit; Khan, Farha; Suguna, Kaza

    2017-09-11

    In response to the widespread presence of inorganic Hg in the environment, bacteria have evolved resistance systems with mercuric reductase (MerA) as the key enzyme. MerA enzymes have still not been well characterized from gram positive bacteria. Current study reports physico-chemical, kinetic and structural characterization of MerA from a multiple heavy metal resistant strain of Lysinibacillus sphaericus, and discusses its implications in bioremediation application. The enzyme was homodimeric with subunit molecular weight of about 60 kDa. The Km and Vmax were found to be 32 µM of HgCl2 and 18 units/mg respectively. The enzyme activity was enhanced by β-mercaptoethanol and NaCl up to concentrations of 500 µM and 100 mM respectively, followed by inhibition at higher concentrations. The enzyme showed maximum activity in the pH range of 7-7.5 and temperature range of 25-50 °C, with melting temperature of 67 °C. Cu(2+) exhibited pronounced inhibition of the enzyme with mixed inhibition pattern. The enzyme contained FAD as the prosthetic group and used NADPH as the preferred electron donor, but it showed slight activity with NADH as well. Structural characterization was carried out by circular dichroism spectrophotometry and X-ray crystallography. X-ray confirmed the homodimeric structure of enzyme and gave an insight on the residues involved in catalytic binding. In conclusion, the investigated enzyme showed higher catalytic efficiency, temperature stability and salt tolerance as compared to MerA enzymes from other mesophiles. Therefore, it is proposed to be a promising candidate for Hg(2+) bioremediation.

  15. Mechanisms involved in the transport of mercuric ions in target tissues.

    PubMed

    Bridges, Christy C; Zalups, Rudolfs K

    2017-01-01

    Mercury exists in the environment in various forms, all of which pose a risk to human health. Despite guidelines regulating the industrial release of mercury into the environment, humans continue to be exposed regularly to various forms of this metal via inhalation or ingestion. Following exposure, mercuric ions are taken up by and accumulate in numerous organs, including brain, intestine, kidney, liver, and placenta. In order to understand the toxicological effects of exposure to mercury, a thorough understanding of the mechanisms that facilitate entry of mercuric ions into target cells must first be obtained. A number of mechanisms for the transport of mercuric ions into target cells and organs have been proposed in recent years. However, the ability of these mechanisms to transport mercuric ions and the regulatory features of these carriers have not been characterized completely. The purpose of this review is to summarize the current findings related to the mechanisms that may be involved in the transport of inorganic and organic forms of mercury in target tissues and organs. This review will describe mechanisms known to be involved in the transport of mercury and will also propose additional mechanisms that may potentially be involved in the transport of mercuric ions into target cells.

  16. A novel mercuric reductase from the unique deep brine environment of Atlantis II in the Red Sea.

    PubMed

    Sayed, Ahmed; Ghazy, Mohamed A; Ferreira, Ari J S; Setubal, João C; Chambergo, Felipe S; Ouf, Amged; Adel, Mustafa; Dawe, Adam S; Archer, John A C; Bajic, Vladimir B; Siam, Rania; El-Dorry, Hamza

    2014-01-17

    A unique combination of physicochemical conditions prevails in the lower convective layer (LCL) of the brine pool at Atlantis II (ATII) Deep in the Red Sea. With a maximum depth of over 2000 m, the pool is characterized by acidic pH (5.3), high temperature (68 °C), salinity (26%), low light levels, anoxia, and high concentrations of heavy metals. We have established a metagenomic dataset derived from the microbial community in the LCL, and here we describe a gene for a novel mercuric reductase, a key component of the bacterial detoxification system for mercuric and organomercurial species. The metagenome-derived gene and an ortholog from an uncultured soil bacterium were synthesized and expressed in Escherichia coli. The properties of their products show that, in contrast to the soil enzyme, the ATII-LCL mercuric reductase is functional in high salt, stable at high temperatures, resistant to high concentrations of Hg(2+), and efficiently detoxifies Hg(2+) in vivo. Interestingly, despite the marked functional differences between the orthologs, their amino acid sequences differ by less than 10%. Site-directed mutagenesis and kinetic analysis of the mutant enzymes, in conjunction with three-dimensional modeling, have identified distinct structural features that contribute to extreme halophilicity, thermostability, and high detoxification capacity, suggesting that these were acquired independently during the evolution of this enzyme. Thus, our work provides fundamental structural insights into a novel protein that has undergone multiple biochemical and biophysical adaptations to promote the survival of microorganisms that reside in the extremely demanding environment of the ATII-LCL.

  17. A Novel Mercuric Reductase from the Unique Deep Brine Environment of Atlantis II in the Red Sea

    PubMed Central

    Sayed, Ahmed; Ghazy, Mohamed A.; Ferreira, Ari J. S.; Setubal, João C.; Chambergo, Felipe S.; Ouf, Amged; Adel, Mustafa; Dawe, Adam S.; Archer, John A. C.; Bajic, Vladimir B.; Siam, Rania; El-Dorry, Hamza

    2014-01-01

    A unique combination of physicochemical conditions prevails in the lower convective layer (LCL) of the brine pool at Atlantis II (ATII) Deep in the Red Sea. With a maximum depth of over 2000 m, the pool is characterized by acidic pH (5.3), high temperature (68 °C), salinity (26%), low light levels, anoxia, and high concentrations of heavy metals. We have established a metagenomic dataset derived from the microbial community in the LCL, and here we describe a gene for a novel mercuric reductase, a key component of the bacterial detoxification system for mercuric and organomercurial species. The metagenome-derived gene and an ortholog from an uncultured soil bacterium were synthesized and expressed in Escherichia coli. The properties of their products show that, in contrast to the soil enzyme, the ATII-LCL mercuric reductase is functional in high salt, stable at high temperatures, resistant to high concentrations of Hg2+, and efficiently detoxifies Hg2+ in vivo. Interestingly, despite the marked functional differences between the orthologs, their amino acid sequences differ by less than 10%. Site-directed mutagenesis and kinetic analysis of the mutant enzymes, in conjunction with three-dimensional modeling, have identified distinct structural features that contribute to extreme halophilicity, thermostability, and high detoxification capacity, suggesting that these were acquired independently during the evolution of this enzyme. Thus, our work provides fundamental structural insights into a novel protein that has undergone multiple biochemical and biophysical adaptations to promote the survival of microorganisms that reside in the extremely demanding environment of the ATII-LCL. PMID:24280218

  18. Mercuric reductase activity and evidence of broad-spectrum mercury resistance among clinical isolates of rapidly growing mycobacteria

    SciTech Connect

    Steingrube, V.A.; Wallace, R.J. Jr.; Steele, L.C.; Pang, Y.J. )

    1991-05-01

    Resistance to mercury was evaluated in 356 rapidly growing mycobacteria belonging to eight taxonomic groups. Resistance to inorganic Hg2+ ranged from 0% among the unnamed third biovariant complex of Mycobacterium fortuitum to 83% among M. chelonae-like organisms. With cell extracts and 203Hg(NO3)2 as the substrate, mercuric reductase (HgRe) activity was demonstrable in six of eight taxonomic groups. HgRe activity was inducible and required NADPH or NADH and a thiol donor for optimai activity. Species with HgRe activity were also resistant to organomercurial compounds, including phenylmercuric acetate. Attempts at intraspecies and intragenus transfer of HgRe activity by conjugation or transformation were unsuccessful. Mercury resistance is common in rapidly growing mycobacteria and appears to function via the same inducible enzyme systems already defined in other bacterial species. This system offers potential as a strain marker for epidemiologic investigations and for studying genetic systems in rapidly growing mycobacteria.

  19. Evidence for the participation of Cys sub 558 and Cys sub 559 at the active site of mercuric reductase

    SciTech Connect

    Miller, S.M.; Moore, M.J.; Massey, V.; Williams, C.H. Jr.; Distefano, M.D.; Ballou, D.P.; Walsh, C.T. )

    1989-02-07

    Mercuric reductase, with FAD and a reducible disulfide at the active site, catalyzes the two-electron reduction of Hg(II) by NADPH. Addition of reducing equivalents rapidly produces a spectrally distinct EH{sub 2} form of the enzyme containing oxidized FAD and reduced active site thiols. Formation of EH{sub 2} has previously been reported to require only 2 electrons for reduction of the active site disulfide. The authors present results of anaerobic titrations of mercuric reductase with NADPH and dithionite showing that the equilibrium conversion of oxidized enzyme to EH{sub 2} actually requires 2 equiv of reducing agent or 4 electrons. Kinetic studies conducted both at 4{degree}C and at 25{degree}C indicate that reduction of the active site occurs rapidly, as previously reported; this is followed by a slower reduction of another redox group via reaction with the active site. ({sup 14}C)Iodoacetamide labeling experiments demonstrate that the C-terminal residues, Cys{sub 558} and Cys{sub 559}, are involved in this disulfide. The fluorescence, but not the absorbance, of the enzyme-bound FAD was found to be highly dependent on the redox state of the C-terminal thiols. Thus, E{sub ox} with Cys{sub 558} and Cys{sub 559} as thiols exhibits less than 50% of the fluorescence of E{sub ox} where these residues are present as a disulfide, indicating that the thiols remain intimately associated with the active site. Initial velocity measurements show that the auxiliary disulfide must be reduced before catalytic Hg(II) reduction can occur, consistent with the report of a preactivation phenomenon with NADPH or cysteine. A modified minimal catalytic mechanism is proposed as well as several chemical mechanisms for the Hg(II) reduction step.

  20. Ultra-low level optical detection of mercuric ions using biogenic gold nanotriangles.

    PubMed

    Singh, Amit; Pasricha, Renu; Sastry, Murali

    2012-07-07

    Mercury is a serious environmental pollutant known to have detrimental health effects in all life forms. Here, we report the use of biologically synthesized aqueous gold nanotriangles for sensitive and selective optical detection of femto-molar levels of mercury ions by exploiting the high amalgamation tendency of mercury metal towards gold. Aqueous chloroaurate ions were reduced using lemongrass (Cymbopogon flexuosus) leaf extract at room temperature to form gold nanotriangles. Mercuric (Hg(2+)) ions were reduced in the presence of these triangles to facilitate amalgamation and the optical properties were monitored. We observe a significant change in the longitudinal plasmon absorption band of the nanotriangles even at femto-molar concentrations of mercuric ions. High-resolution transmission electron microscopy confirms changes in particle morphology at such low concentrations. This protocol shows no sensitivity to other environmentally relevant metal ions, including Pb(2+), Zn(2+), Cd(2+), Fe(2+), Ni(2+), Sr(2+), Ca(2+), Mn(2+), and Cu(2+), confirming further that change in the optical properties of gold nanotriangles in the presence of reduced mercuric ions is solely due to the strong amalgamation tendency of mercury metal.

  1. Genes encoding mercuric reductases from selected gram-negative aquatic bacteria have a low degree of homology with merA of transposon Tn501.

    PubMed Central

    Barkay, T; Gillman, M; Liebert, C

    1990-01-01

    An investigation of the Hg2+ resistance mechanism of four freshwater and four coastal marine bacteria that did not hybridize with a mer operonic probe was conducted (T. Barkay, C. Liebert, and M. Gillman, Appl. Environ. Microbiol. 55:1196-1202, 1989). Hybridization with a merA probe, the gene encoding the mercuric reductase polypeptide, at a stringency of hybridization permitting hybrid formation between evolutionarily distant merA genes (as exists between gram-positive and -negative bacteria), detected merA sequences in the genomes of all tested strains. Inducible Hg2+ volatilization was demonstrated for all eight organisms, and NADPH-dependent mercuric reductase activities were detected in crude cell extracts of six of the strains. Because these strains represented random selections of bacteria from three aquatic environments, it is concluded that merA encodes a common molecular mechanism for Hg2+ resistance and volatilization in aerobic heterotrophic aquatic communities. Images PMID:2166470

  2. Effect of mercuric ion on attraction to light of artemia sp nauplii.

    PubMed

    Saunders, J P; Trieff, N M; Kalmaz, E E; Uchida, T

    1985-02-01

    Living organisms exhibit a phototactic response which can be altered by certain environmental toxic chemical species. The analysis of photobehavior can help in elucidating environmental factors that influence photomotility reactions of the organisms. A method has been developed that measures the phototactic response of Artemia nauplii under the influence of mercuric ion (Hg2+) in synthetic seawater. The phototactic response of Artemia nauplii was manifested by movement of the organisms from a darkened half to lighted half of an experimental vessel containing synthetic seawater. The density as a function of time of Artemia nauplii is determined by removing aliquots from both light and dark sides and then plating on agar for counting under the dissecting microscope. Measurements consistently show a significant movement of nauplii to the lighted side within 45 min of the start of the experiments. The present investigation demonstrated that at concentrations as low as 0.010 mg HgCl2/liter there is an enhancement of phototactic effect on Artemia nauplii by mercuric ion as compared with control. The phototactic response of Artema nauplii is altered by mercuric ion in a dose-related manner, but the mechanism of this effect is presently unknown.

  3. Mercuric reductase genes (merA) and mercury resistance plasmids in High Arctic snow, freshwater and sea-ice brine.

    PubMed

    Møller, Annette K; Barkay, Tamar; Hansen, Martin A; Norman, Anders; Hansen, Lars H; Sørensen, Søren J; Boyd, Eric S; Kroer, Niels

    2014-01-01

    Bacterial reduction in Hg(2+) to Hg(0) , mediated by the mercuric reductase (MerA), is important in the biogeochemical cycling of Hg in temperate environments. Little is known about the occurrence and diversity of merA in the Arctic. Seven merA determinants were identified among bacterial isolates from High Arctic snow, freshwater and sea-ice brine. Three determinants in Bacteriodetes, Firmicutes and Actinobacteria showed < 92% (amino acid) sequence similarity to known merA, while one merA homologue in Alphaproteobacteria and 3 homologues from Betaproteobacteria and Gammaproteobacteria were > 99% similar to known merA's. Phylogenetic analysis showed the Bacteroidetes merA to be part of an early lineage in the mer phylogeny, whereas the Betaproteobacteria and Gammaproteobacteria merA appeared to have evolved recently. Several isolates, in which merA was not detected, were able to reduce Hg(2+) , suggesting presence of unidentified merA genes. About 25% of the isolates contained plasmids, two of which encoded mer operons. One plasmid was a broad host-range IncP-α plasmid. No known incompatibility group could be assigned to the others. The presence of conjugative plasmids, and an incongruent distribution of merA within the taxonomic groups, suggests horizontal transfer of merA as a likely mechanism for High Arctic microbial communities to adapt to changing mercury concentration.

  4. Triggered activity of a nicking endonuclease for mercuric(II) ion-mediated duplex-like DNA cleavage.

    PubMed

    Li, Feng; Feng, Yan; Liu, Shufeng; Tang, Bo

    2011-06-14

    The cleavage activity of a nicking endonuclease towards metal-ion-mediated duplex-like DNA can be triggered by the corresponding metal ions, which was demonstrated with mercuric(II) ion as a model via a simple electrochemical protocol. This journal is © The Royal Society of Chemistry 2011

  5. Mercuric ion reduction and resistance in transgenic Arabidopsis thaliana plants expressing a modified bacterial merA gene.

    PubMed Central

    Rugh, C L; Wilde, H D; Stack, N M; Thompson, D M; Summers, A O; Meagher, R B

    1996-01-01

    With global heavy metal contamination increasing, plants that can process heavy metals might provide efficient and ecologically sound approaches to sequestration and removal. Mercuric ion reductase, MerA, converts toxic Hg2+ to the less toxic, relatively inert metallic mercury (Hg0) The bacterial merA sequence is rich in CpG dinucleotides and has a highly skewed codon usage, both of which are particularly unfavorable to efficient expression in plants. We constructed a mutagenized merA sequence, merApe9, modifying the flanking region and 9% of the coding region and placing this sequence under control of plant regulatory elements. Transgenic Arabidopsis thaliana seeds expressing merApe9 germinated, and these seedlings grew, flowered, and set seed on medium containing HgCl2 concentrations of 25-100 microM (5-20 ppm), levels toxic to several controls. Transgenic merApe9 seedlings evolved considerable amounts of Hg0 relative to control plants. The rate of mercury evolution and the level of resistance were proportional to the steady-state mRNA level, confirming that resistance was due to expression of the MerApe9 enzyme. Plants and bacteria expressing merApe9 were also resistant to toxic levels of Au3+. These and other data suggest that there are potentially viable molecular genetic approaches to the phytoremediation of metal ion pollution. Images Fig. 2 Fig. 3 Fig. 4 PMID:8622910

  6. High diversity of bacterial mercuric reductase genes from surface and sub-surface floodplain soil (Oak Ridge, USA).

    PubMed

    Oregaard, Gunnar; Sørensen, Søren J

    2007-09-01

    DNA was extracted from different depth soils (0-5, 45-55 and 90-100 cm below surface) sampled at Lower East Fork Poplar Creek floodplain (LEFPCF), Oak Ridge (TN, USA). The presence of merA genes, encoding the mercuric reductase, the key enzyme in detoxification of mercury in bacteria, was examined by PCR targeting Actinobacteria, Firmicutes or beta/gamma-Proteobacteria. beta/gamma-Proteobacteria merA genes were successfully amplified from all soils, whereas Actinobacteria were amplified only from surface soil. merA clone libraries were constructed and sequenced. beta/gamma-Proteobacteria sequences revealed high diversity in all soils, but limited vertical similarity. Less than 20% of the operational taxonomic units (OTU) (DNA sequences > or = 95% identical) were shared between the different soils. Only one of the 62 OTU was > or = 95% identical to a GenBank sequence, highlighting that cultivated bacteria are not representative of what is found in nature. Fewer merA sequences were obtained from the Actinobacteria, but these were also diverse, and all were different from GenBank sequences. A single clone was most closely related to merA of alpha-Proteobacteria. An alignment of putative merA genes of genome sequenced mainly marine alpha-Proteobacteria was used for design of merA primers. PCR amplification of soil alpha-Proteobacteria isolates and sequencing revealed that they were very different from the genome-sequenced bacteria (only 62%-66% identical at the amino-acid level), although internally similar. In light of the high functional diversity of mercury resistance genes and the limited vertical distribution of shared OTU, we discuss the role of horizontal gene transfer as a mechanism of bacterial adaptation to mercury.

  7. Highly photoluminescent silicon nanocrystals for rapid, label-free and recyclable detection of mercuric ions

    NASA Astrophysics Data System (ADS)

    Zhang, Jia; Yu, Shu-Hong

    2014-03-01

    Hydrothermal treatment of 3-aminopropyltrimethoxysilane (APTMS) in the presence of sodium citrate generates a suspension of highly fluorescent silicon nanocrystals that fluoresces blue under UV irradiation. The photoluminescent quantum yield of the as-prepared silicon nanocrystals was calculated to be 21.6%, with quinine sulfate as the standard reference. Only mercuric ions (Hg2+) can readily prevent the fluorescence of the silicon nanocrystals, indicating a remarkably high selectivity towards Hg2+ over other metal ions. The optimized sensor system shows a sensitive detection range from 50 nM to 1 μM and a detection limit of 50 nM. The quenching mechanism was explained in terms of optical absorption spectra and time-resolved fluorescence decay spectra. Due to the strong interaction of Hg2+ with the thiol group, the fluorescence can be fully recovered by biothiols such as cysteine and glutathione, therefore, a regenerative strategy has been proposed and successfully applied to detect Hg2+ by the same sensor for at least five cycles. Endowed with relatively high sensitivity and selectivity, the present sensor holds the potential to be applied for mercuric assay in water.Hydrothermal treatment of 3-aminopropyltrimethoxysilane (APTMS) in the presence of sodium citrate generates a suspension of highly fluorescent silicon nanocrystals that fluoresces blue under UV irradiation. The photoluminescent quantum yield of the as-prepared silicon nanocrystals was calculated to be 21.6%, with quinine sulfate as the standard reference. Only mercuric ions (Hg2+) can readily prevent the fluorescence of the silicon nanocrystals, indicating a remarkably high selectivity towards Hg2+ over other metal ions. The optimized sensor system shows a sensitive detection range from 50 nM to 1 μM and a detection limit of 50 nM. The quenching mechanism was explained in terms of optical absorption spectra and time-resolved fluorescence decay spectra. Due to the strong interaction of Hg2+ with the

  8. Magnetic detection of mercuric ion using giant magnetoresistance-based biosensing system.

    PubMed

    Wang, Wei; Wang, Yi; Tu, Liang; Klein, Todd; Feng, Yinglong; Li, Qin; Wang, Jian-Ping

    2014-04-15

    We have demonstrated a novel sensing strategy employing a giant magnetoresistance (GMR) biosensor and DNA chemistry for the detection of mercuric ion (Hg(2+)). This assay takes advantages of high sensitivity and real-time signal readout of GMR biosensor and high selectivity of thymine-thymine (T-T) pair for Hg(2+). The assay has a detection limit of 10 nM in both buffer and natural water, which is the maximum mercury level in drinking water regulated by U.S. Environmental Protection Agency (EPA). The magnitude of the dynamic range for Hg(2+) detection is up to three orders (10 nM to 10 μM). Herein, GMR sensing technology is first introduced into a pollutant monitoring area. It can be foreseen that the GMR biosensor could become a robust contender in the areas of environmental monitoring and food safety testing.

  9. Highly selective and sensitive detection of mercuric ion based on a visual fluorescence method.

    PubMed

    Yuan, Chao; Zhang, Kui; Zhang, Zhongping; Wang, Suhua

    2012-11-20

    The instant and on-site detection of trace aqueous mercuric ion still remains a challenge for environmental monitoring and protection. This work demonstrates a new analytical method and its utility for visual detection of aqueous Hg(2+) on the basis of a novel water-soluble CdSe-ZnS quantum dots (QDs) functionalized with a bidentate ligand of 2-hydroxyethyldithiocarbamate (HDTC). The fluorescence of the aqueous HDTC modified QDs (HDTC-QDs) could be selectively and efficiently quenched by Hg(2+) through a surface chelating reaction between HDTC and Hg(2+), and the detection limit was measured to be 1 ppb. Most interestingly, the orange fluorescence of the HDTC-QDs gradually changes to red upon the increasing amount of Hg(2+) added besides the decreasing of the fluorescence intensity. By taking advantage of this optical phenomenon, a paper-based sensor for aqueous Hg(2+) detection has been developed by immobilizing the HDTC-QDs on cellulose acetate paper which has low background fluorescence in the wavelength range. The paper-based sensor showed high sensitivity and selectivity for Hg(2+) visual detection. When Hg(2+) was dropped onto the paper-sensor, an obviously distinguishable fluorescence color evolution (from orange to red) could be clearly observed depending on the concentration of Hg(2+). The limit of detection of the visual method for aqueous Hg(2+) detection was as low as 0.2 ppm. The very simple and effective strategy reported here should facilitate the development of portable and reliable fluorescence chemosensors for mercuric pollution control.

  10. A rapid electrochemical procedure for the detection of Hg(0) produced by mercuric-reductase: application for monitoring Hg-resistant bacteria activity.

    PubMed

    Battistel, Dario; Baldi, Franco; Marchetto, Davide; Gallo, Michele; Daniele, Salvatore

    2012-10-02

    In this work, gold microelectrodes are employed as traps for the detection of volatilized metallic mercury produced by mercuric reductase (MerA) extracted from an Hg-resistant Pseudomonas putida strain FB1. The enzymatic reduction of Hg (II) to Hg (0) was induced by NADPH cofactor added to the samples. The amount of Hg(0) accumulated on the gold microelectrode surface was determined by anodic stripping voltammetry (ASV) after transferring the gold microelectrode in an aqueous solution containing 0.1 M HNO(3) + 1 M KNO(3). Electrochemical measurements were combined with spectrofluorometric assays of NADPH consumption to derive an analytical expression for the detection of a relative MerA activity of different samples with respect to that of P. putida. The method developed here was employed for the rapid determination of MerA produced by bacteria harbored in soft tissues of clams (Ruditapes philippinarum), collected in high Hg polluted sediments of Northern Adriatic Sea in Italy.

  11. Femtomolar detection of mercuric ions using polypyrrole, pectin and graphene nanocomposites modified electrode.

    PubMed

    Arulraj, Abraham Daniel; Devasenathipathy, Rajkumar; Chen, Shen-Ming; Vasantha, Vairathevar Sivasamy; Wang, Sea-Fue

    2016-12-01

    Several nanomaterials and techniques for the detection of mercuric ions (Hg(2+)) have been developed in the past decade. However, simple, low-cost and rapid sensor for the detection of heavy metal ions yet remains an important task. Herein, we present a highly sensitive electrochemical sensor for the femtomolar detection of Hg(2+) based on polypyrrole, pectin, and graphene (PPy/Pct/GR) which was prepared by one step electrochemical potentiodyanamic method. The effect of concentration of pectin, polypyrrole and graphene were studied for the detection of Hg(2+). The influence of experimental parameters including effect of pH, accumulation time and accumulation potential were also studied. Different pulse anodic stripping voltammetry was chosen to detect Hg(2+) at PPy/Pct/GR/GCE modified electrode. The fabricated sensor achieved an excellent performance towards Hg(2+) detection such as higher sensitivity of 28.64μAμM(-1) and very low detection limit (LOD) of 4 fM at the signal to noise ratio of 3. The LOD of our sensor offered nearly 6 orders of magnitude lower than that of recommended concentration of Hg(2+) in drinking water by United States Environmental Protection Agency and World Health Organization. Compared to all previously reported electrochemical sensors towards Hg(2+) detection, our newly fabricated sensor attained a very LOD in the detection of Hg(2+). The practicality of our proposed sensor for the detection of Hg(2+) was successfully demonstrated in untreated tap water.

  12. L-cysteine protected copper nanoparticles as colorimetric sensor for mercuric ions.

    PubMed

    Soomro, Razium A; Nafady, Ayman; Sirajuddin; Memon, Najma; Sherazi, Tufail H; Kalwar, Nazar H

    2014-12-01

    This report demonstrates a novel, simple and efficient protocol for the synthesis of copper nanoparticles in aqueous solution using L-cysteine as capping or protecting agent. UV-visible (UV-vis) spectroscopy was employed to monitor the LSPR band of L-cysteine functionalized copper nanoparticles (Cyst-Cu NPs) based on optimizing various reaction parameters. Fourier Transform Infrared (FTIR) spectroscopy provided information about the surface interaction between L-cysteine and Cu NPs. Transmission Electron Microscopy (TEM) confirmed the formation of fine spherical, uniformly distributed Cyst-Cu NPs with average size of 34 ± 2.1 nm. X-ray diffractometry (XRD) illustrated the formation of pure metallic phase crystalline Cyst-Cu NPs. As prepared Cyst-Cu NPs were tested as colorimetric sensor for determining mercuric (Hg(2+)) ions in an aqueous system. Cyst-Cu NPs demonstrated very sensitive and selective colorimetric detection of Hg(2+) ions in the range of 0.5 × 10(-6)-3.5 × 10(-6) mol L(-1) based on decrease in LSPR intensity as monitored by a UV-vis spectrophotometer. The developed sensor is simple, economic compared to those based on precious metal nanoparticles and sensitive to detect Hg(2+) ions with detection limit down to 4.3 × 10(-8) mol L(-1). The sensor developed in this work has a high potential for rapid and on-site detection of Hg(2+) ions. The sensor was successfully applied for assessment of Hg(2+) ions in real water samples collected from various locations of the Sindh River. Copyright © 2014 Elsevier B.V. All rights reserved.

  13. Intermingled modulatory and neurotoxic effects of thimerosal and mercuric ions on electrophysiological responses to GABA and NMDA in hippocampal neurons.

    PubMed

    Wyrembek, P; Szczuraszek, K; Majewska, M D; Mozrzymas, J W

    2010-12-01

    The organomercurial, thimerosal, is at the center of medical controversy as a suspected factor contributing to neurodevelopmental disorders in children. Many neurotoxic effects of thimerosal have been described, but its interaction with principal excitatory and inhibitory neurotransmiter systems is not known. We examined, using electrophysiological recordings, thimerosal effects on GABA and NMDA-evoked currents in cultured hippocampal neurons. After brief (3 to 10 min) exposure to thimerosal at concentrations up to 100 μM, there was no significant effect on GABA or NMDA-evoked currents. However, following exposure for 60-90 min to 1 or 10 μM thimerosal, there was a significant decrease in NMDA-induced currents (p<0.05) and GABAergic currents (p<0.05). Thimerosal was also neurotoxic, damaging a significant proportion of neurons after 60-90 min exposure; recordings were always conducted in the healthiest looking neurons. Mercuric chloride, at concentrations 1 μM and above, was even more toxic, killing a large proportion of cells after just a few minutes of exposure. Recordings from a few sturdy cells revealed that micromolar mercuric chloride markedly potentiated the GABAergic currents (p<0.05), but reduced NMDA-evoked currents (p<0.05). The results reveal complex interactions of thimerosal and mercuric ions with the GABA(A) and NMDA receptors. Mercuric chloride act rapidly, decreasing electrophysiological responses to NMDA but enhancing responses to GABA, while thimerosal works slowly, reducing both NMDA and GABA responses. The neurotoxic effects of both mercurials are interwoven with their modulatory actions on GABA(A) and NMDA receptors, which most likely involve binding to these macromolecules.

  14. MRP2 and the Handling of Mercuric Ions in Rats Exposed Acutely to Inorganic and Organic Species of Mercury

    PubMed Central

    Bridges, Christy C.; Joshee, Lucy; Zalups, Rudolfs K.

    2011-01-01

    Mercuric ions accumulate preferentially in renal tubular epithelial cells and bond with intracellular thiols. Certain metal-complexing agents have been shown to promote extraction of mercuric ions via the multidrug resistance-associated protein 2 (MRP2). Following exposure to a non-toxic dose of inorganic mercury (Hg2+), in the absence of complexing agents, tubular cells are capable of exporting a small fraction of intracellular Hg2+ through one or more undetermined mechanisms. We hypothesize that MRP2 plays a role in this export. To test this hypothesis, Wistar (control) and TR− rats were injected intravenously with a non-nephrotoxic dose of HgCl2 (0.5 μmol/kg) or CH3HgCl (5 mg/kg), containing [203Hg], in the presence or absence of cysteine (Cys; 1.25 μmol/kg or 12.5 mg/kg, respectively). Animals were sacrificed 24 h after exposure to mercury and the content of [203Hg] in blood, kidneys, liver, urine and feces was determined. In addition, uptake of Cys-S-conjugates of Hg2+ and methylmercury (CH3Hg+) was measured in inside-out membrane vesicles prepared from either control Sf9 cells or Sf9 cells transfected with human MRP2. The amount of mercury in the total renal mass and liver was significantly greater in TR− rats than in controls. In contrast, the amount of mercury in urine and feces was significantly lower in TR− rats than in controls. Data from membrane vesicles indicate that Cys-S-conjugates of Hg2+ and CH3Hg+ are transportable substrates of MRP2. Collectively, these data indicate that MRP2 plays a role in the physiological handling and elimination of mercuric ions from the kidney. PMID:21134393

  15. MRP2 and the handling of mercuric ions in rats exposed acutely to inorganic and organic species of mercury

    SciTech Connect

    Bridges, Christy C. Joshee, Lucy; Zalups, Rudolfs K.

    2011-02-15

    Mercuric ions accumulate preferentially in renal tubular epithelial cells and bond with intracellular thiols. Certain metal-complexing agents have been shown to promote extraction of mercuric ions via the multidrug resistance-associated protein 2 (MRP2). Following exposure to a non-toxic dose of inorganic mercury (Hg{sup 2+}), in the absence of complexing agents, tubular cells are capable of exporting a small fraction of intracellular Hg{sup 2+} through one or more undetermined mechanisms. We hypothesize that MRP2 plays a role in this export. To test this hypothesis, Wistar (control) and TR{sup -} rats were injected intravenously with a non-nephrotoxic dose of HgCl{sub 2} (0.5 {mu}mol/kg) or CH{sub 3}HgCl (5 mg/kg), containing [{sup 203}Hg], in the presence or absence of cysteine (Cys; 1.25 {mu}mol/kg or 12.5 mg/kg, respectively). Animals were sacrificed 24 h after exposure to mercury and the content of [{sup 203}Hg] in blood, kidneys, liver, urine and feces was determined. In addition, uptake of Cys-S-conjugates of Hg{sup 2+} and methylmercury (CH{sub 3}Hg{sup +}) was measured in inside-out membrane vesicles prepared from either control Sf9 cells or Sf9 cells transfected with human MRP2. The amount of mercury in the total renal mass and liver was significantly greater in TR{sup -} rats than in controls. In contrast, the amount of mercury in urine and feces was significantly lower in TR{sup -} rats than in controls. Data from membrane vesicles indicate that Cys-S-conjugates of Hg{sup 2+} and CH{sub 3}Hg{sup +} are transportable substrates of MRP2. Collectively, these data indicate that MRP2 plays a role in the physiological handling and elimination of mercuric ions from the kidney.

  16. Toxicological Significance of Renal Bcrp: Another Potential Transporter in the Elimination of Mercuric Ions from Proximal Tubular Cells

    PubMed Central

    Bridges, Christy C.; Zalups, Rudolfs K.; Joshee, Lucy

    2015-01-01

    Secretion of inorganic mercury (Hg2+) from proximal tubular cells into the tubular lumen has been shown to involve the multidrug resistance-associated protein 2 (Mrp2). Considering similarities in localization and substrate specificity between Mrp2 and the breast cancer resistance protein (Bcrp), we hypothesize that Bcrp may also play a role in the proximal tubular secretion of mercuric species. In order to test this hypothesis, the uptake of Hg2+ was examined initially using inside-out membrane vesicles containing Bcrp. The results of these studies suggest that Bcrp may be capable of transporting certain conjugates of Hg2+. To further characterize the role of Bcrp in the handling of mercuric ions and in the induction of Hg2+-induced nephropathy, Sprague-Dawley and Bcrp knockout (bcrp−/−) rats were exposed intravenously to a non-nephrotoxic (0.5 μmol • kg−1), a moderately nephrotoxic (1.5 μmol • kg−1) or a significantly nephrotoxic (2.0 μmol • kg−1) dose of HgCl2. In general, the accumulation of Hg2+ was greater in organs of bcrp−/− rats than in Sprague-Dawley rats, suggesting that Bcrp may play a role in the export of Hg2+ from target cells. Within the kidney, cellular injury and necrosis was more severe in bcrp−/− rats than in controls. The pattern of necrosis, which was localized in the inner cortex and the outer stripe of the outer medulla was significantly different from that observed in Mrp2-deficient animals. These findings suggest that Bcrp may be involved in the cellular export of select mercuric species and that its role in this export may differ from that of Mrp2. PMID:25868844

  17. Tn5563, a transposon encoding putative mercuric ion transport proteins located on plasmid pRA2 of Pseudomonas alcaligenes.

    PubMed

    Yeo, C C; Tham, J M; Kwong, S M; Yiin, S; Poh, C L

    1998-08-15

    Sequence analysis of pRA2, an endogenous 33-kb plasmid from Pseudomonas alcaligenes NCIB 9867 (strain P25X), revealed the presence of a 6256-bp transposon of the Tn3 family, designated Tn5563. Tn5563, which is flanked by two 39-bp inverted repeats, encodes a transposase, a resolvase, and two open reading frames which share amino acid sequence similarities with the mercuric ion transport proteins MerT and MerP encoded by several mer operons. However, no other mer operon genes were found on Tn5563. Sequencing of a RP4::XIn hybrid plasmid indicates possible interactions between pRA2 and the P25X chromosome mediated by Tn5563.

  18. Toxicological significance of renal Bcrp: Another potential transporter in the elimination of mercuric ions from proximal tubular cells

    SciTech Connect

    Bridges, Christy C. Zalups, Rudolfs K.; Joshee, Lucy

    2015-06-01

    Secretion of inorganic mercury (Hg{sup 2+}) from proximal tubular cells into the tubular lumen has been shown to involve the multidrug resistance-associated protein 2 (Mrp2). Considering similarities in localization and substrate specificity between Mrp2 and the breast cancer resistance protein (Bcrp), we hypothesize that Bcrp may also play a role in the proximal tubular secretion of mercuric species. In order to test this hypothesis, the uptake of Hg{sup 2+} was examined initially using inside-out membrane vesicles containing Bcrp. The results of these studies suggest that Bcrp may be capable of transporting certain conjugates of Hg{sup 2+}. To further characterize the role of Bcrp in the handling of mercuric ions and in the induction of Hg{sup 2+}-induced nephropathy, Sprague–Dawley and Bcrp knockout (bcrp{sup −/−}) rats were exposed intravenously to a non-nephrotoxic (0.5 μmol·kg{sup −1}), a moderately nephrotoxic (1.5 μmol·kg{sup −1}) or a significantly nephrotoxic (2.0 μmol·kg{sup −1}) dose of HgCl{sub 2}. In general, the accumulation of Hg{sup 2+} was greater in organs of bcrp{sup −/−} rats than in Sprague–Dawley rats, suggesting that Bcrp may play a role in the export of Hg{sup 2+} from target cells. Within the kidney, cellular injury and necrosis was more severe in bcrp{sup −/−} rats than in controls. The pattern of necrosis, which was localized in the inner cortex and the outer stripe of the outer medulla, was significantly different from that observed in Mrp2-deficient animals. These findings suggest that Bcrp may be involved in the cellular export of select mercuric species and that its role in this export may differ from that of Mrp2. - Highlights: • Bcrp may mediate transport of mercury out of proximal tubular cells. • Hg-induced nephropathy was more severe in Bcrp knockout rats. • Bcrp and Mrp2 may differ in their ability to transport Hg.

  19. Effect of ambient conditions on simultaneous growth and bioaccumulation of mercuric ion by genetically engineered E. coli JM109.

    PubMed

    Deng, Xu; Zheng, Yangchun; Li, Qingbiao

    2006-08-21

    Genetically engineered E. coli JM109, namely M1, which expressed both Hg(2+) transport system and metallothionein, was tested for its capability of simultaneous growth and bioaccumulation of Hg(2+) under low nutritional circumstances. The influential factors of ambient conditions, e.g. initial concentrations of mercuric ion, ionic strength, the presence of metal chelators and other coexisting metal ions were investigated. Hg(2+) bioaccumulation behavior of M1 proved to be well coupled with its growth. NaCl was essential to the growth of M1. Of all tested NaCl concentrations, 0.04 mol/L was optimal. The presence of 0.1 mol/L CaCl(2) or MgCl(2) could promote the growth of M1 and keep the Hg(2+) removal ratio high, but the growth of M1 was inhibited seriously as the concentration of CaCl(2) or MgCl(2) reached 0.3 mol/L. Chelator EDTA had a significant influence on M1 growth and Hg(2+) bioaccumulation, while the effect of citration was little. The presence of other coexisting metal ions inhibited the growth of M1. The influential order was as follows: Cd(2+)>Zn(2+)> or =Cu(2+)>Pb(2+)>Ni(2+). However, only Cd(2+) and Cu(2+) posed obviously adverse effects on Hg(2+) bioaccumulation during the SG&B process.

  20. Mercuric ions inhibit mitogen-activated protein kinase dephosphorylation by inducing reactive oxygen species

    SciTech Connect

    Haase, Hajo; Engelhardt, Gabriela; Hebel, Silke; Rink, Lothar

    2011-01-01

    Mercury intoxication profoundly affects the immune system, in particular, signal transduction of immune cells. However, the mechanism of the interaction of mercury with cellular signaling pathways, such as mitogen activated protein kinases (MAPK), remains elusive. Therefore, the objective of this study is to investigate three potential ways in which Hg{sup 2+} ions could inhibit MAPK dephosphorylation in the human T-cell line Jurkat: (1) by direct binding to phosphatases; (2) by releasing cellular zinc (Zn{sup 2+}); and (3) by inducing reactive oxygen species (ROS). Hg{sup 2+} causes production of ROS, measured by dihydrorhodamine 123, and triggers ROS-mediated Zn{sup 2+} release, detected with FluoZin-3. Yet, phosphatase-inhibition is not mediated by binding of Zn{sup 2+} or Hg{sup 2+}. Rather, phosphatases are inactivated by at least two forms of thiol oxidation; initial inhibition is reversible with reducing agents such as Tris(2-carboxyethyl)phosphine. Prolonged inhibition leads to non-reversible phosphatase oxidation, presumably oxidizing the cysteine thiol to sulfinic- or sulfonic acid. Notably, phosphatases are a particularly sensitive target for Hg{sup 2+}-induced oxidation, because phosphatase activity is inhibited at concentrations of Hg{sup 2+} that have only minor impact on over all thiol oxidation. This phosphatase inhibition results in augmented, ROS-dependent MAPK phosphorylation. MAPK are important regulators of T-cell function, and MAPK-activation by inhibition of phosphatases seems to be one of the molecular mechanisms by which mercury affects the immune system.

  1. Mercuric ion-resistance operons of plasmid R100 and transposon Tn501: the beginning of the operon including the regulatory region and the first two structural genes.

    PubMed Central

    Misra, T K; Brown, N L; Fritzinger, D C; Pridmore, R D; Barnes, W M; Haberstroh, L; Silver, S

    1984-01-01

    The mercuric ion-resistance operons of plasmid R100 (originally from Shigella) and transposon Tn501 (originally from a plasmid isolated in Pseudomonas) have been compared by DNA sequence analysis. The sequences for the first 1340 base pairs of Tn501 are given with the best alignment with the comparable 1319 base pairs of R100. The homology between the two sequences starts at base 58 after the end of the insertion sequence IS-1 of R100. The sequences include the transcriptional regulatory region, and the homology is particularly strong in regions just upstream from potential transcriptional initiation sites. The trans-acting regulatory gene merR consists of 180 base pairs in both cases and codes for a highly basic polypeptide of 60 amino acids, which is also rich in serine. The Tn501 and R100 merR genes differ in 25 of the 180 base positions, and the resulting polypeptides differ in seven amino acids. The regulatory region before the major transcription initiation site contains potential -35 and -10 sequences and dyad symmetrical sequences, which may be the merR binding sites for transcriptional regulation. The first structural gene, merT, encodes a highly hydrophobic polypeptide of 116 amino acids. The R100 and Tn501 merT genes differ in 17% of their positions, leading to 14 (12%) amino acid changes. This region had previously been shown to encode a protein governing membrane transport of mercuric ions. The second structural gene, merC, would give a 91 amino acid polypeptide with a hydrophobic amino-terminal segment. The Tn501 and R100 merC genes differ at 37 base positions, leading to 10 amino acid changes. PMID:6091128

  2. X-ray Structure of a Hg2+ Complex of Mercuric Reductase (MerA) and Quantum Mechanical/Molecular Mechanical Study of Hg2+ Transfer between the C-Terminal and Buried Catalytic Site Cysteine Pairs

    PubMed Central

    2015-01-01

    Mercuric reductase, MerA, is a key enzyme in bacterial mercury resistance. This homodimeric enzyme captures and reduces toxic Hg2+ to Hg0, which is relatively unreactive and can exit the cell passively. Prior to reduction, the Hg2+ is transferred from a pair of cysteines (C558′ and C559′ using Tn501 numbering) at the C-terminus of one monomer to another pair of cysteines (C136 and C141) in the catalytic site of the other monomer. Here, we present the X-ray structure of the C-terminal Hg2+ complex of the C136A/C141A double mutant of the Tn501 MerA catalytic core and explore the molecular mechanism of this Hg transfer with quantum mechanical/molecular mechanical (QM/MM) calculations. The transfer is found to be nearly thermoneutral and to pass through a stable tricoordinated intermediate that is marginally less stable than the two end states. For the overall process, Hg2+ is always paired with at least two thiolates and thus is present at both the C-terminal and catalytic binding sites as a neutral complex. Prior to Hg2+ transfer, C141 is negatively charged. As Hg2+ is transferred into the catalytic site, a proton is transferred from C136 to C559′ while C558′ becomes negatively charged, resulting in the net transfer of a negative charge over a distance of ∼7.5 Å. Thus, the transport of this soft divalent cation is made energetically feasible by pairing a competition between multiple Cys thiols and/or thiolates for Hg2+ with a competition between the Hg2+ and protons for the thiolates. PMID:25343681

  3. X-ray Structure of a Hg2+ Complex of Mercuric Reductase (MerA) and Quantum Mechanical/Molecular Mechanical Study of Hg2+ Transfer between the C-Terminal and Buried Catalytic Site Cysteine Pairs

    DOE PAGES

    Lian, Peng; Guo, Hao-Bo; Riccardi, Demian; ...

    2014-10-24

    Here we report that mercuric reductase, MerA, is a key enzyme in bacterial mercury resistance. This homodimeric enzyme captures and reduces toxic Hg2+ to Hg0, which is relatively unreactive and can exit the cell passively. Prior to reduction, the Hg2+ is transferred from a pair of cysteines (C558' and C559' using Tn501 numbering) at the C-terminus of one monomer to another pair of cysteines (C136 and C141) in the catalytic site of the other monomer. Here, we present the X-ray structure of the C-terminal Hg2+ complex of the C136A/C141A double mutant of the Tn501 MerA catalytic core and explore themore » molecular mechanism of this Hg transfer with quantum mechanical/molecular mechanical (QM/MM) calculations. The transfer is found to be nearly thermoneutral and to pass through a stable tricoordinated intermediate that is marginally less stable than the two end states. For the overall process, Hg2+ is always paired with at least two thiolates and thus is present at both the C-terminal and catalytic binding sites as a neutral complex. Prior to Hg2+ transfer, C141 is negatively charged. As Hg2+ is transferred into the catalytic site, a proton is transferred from C136 to C559' while C558' becomes negatively charged, resulting in the net transfer of a negative charge over a distance of ~7.5 Å. Thus, the transport of this soft divalent cation is made energetically feasible by pairing a competition between multiple Cys thiols and/or thiolates for Hg2+ with a competition between the Hg2+ and protons for the thiolates.« less

  4. Enzymatic detection of mercuric ions in ground-water from vegetable wastes by immobilizing pumpkin (Cucumis melo) urease in calcium alginate beads.

    PubMed

    Prakash, Om; Talat, Mahe; Hasan, Syed Hadi; Pandey, Rajesh K

    2008-07-01

    Present report describes a quick and simple test based on enzyme inhibition for the detection of mercury in aqueous medium by urease immobilized in alginate beads. Urease was extracted from the discarded seeds of pumpkin (Cucumis melo) and was purified to apparent homogeneity (5.2-fold) by heat treatment at 48+/-0.1 degrees C and gel filtration through Sephadex G-200. The homogeneous enzyme preparation (Sp activity 353 U/mg protein, A(280)/A(260)=1.12) was immobilized in 3.5% alginate leading to 86% immobilization. Effect of mercuric ion on the activity of soluble as well as immobilized enzyme was investigated. Hg(2+) exhibited a concentration-dependent inhibition both in the presence and absence of the substrate. The alginate immobilized enzyme showed less inhibition. There was no leaching of the enzyme over a period of 15 days at 4 degrees C. The inhibition was non-competitive and the K(i) was found to be 1.26x10(-1)microM. Time-dependent interaction of urease with Hg(2+) exhibited a biphasic inhibition behavior in which approximately half of the initial activity was lost rapidly (within 10 min) and reminder in a slow phase. Binding of Hg(2+) with the enzyme was largely irreversible, as the activity could not be restored by dialysis. The significance of the observations is discussed.

  5. One-step synthesis of graphitic carbon nitride nanosheets with the help of melamine and its application for fluorescence detection of mercuric ions.

    PubMed

    Zhuang, Qianfen; Sun, Liming; Ni, Yongnian

    2017-03-01

    A facile, simple, and relatively environment-friendly hydrothermal approach was developed for one-step synthesis of graphitic carbon nitride nanosheets (GCNNs) using melamine and sodium citrate as the precursors. The prepared GCNNs emit strong fluorescence with a high quantum yield of 48.3%. The GCNNs were then characterized by various techniques including transmission electron microscopy, atomic force microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and UV-Vis absorption spectroscopy. In addition, the fluorescence quenching behavior of the GCNNS by mercuric ions (Hg(2+)) was exploited to fabricate a label-free fluorescence quenching sensor for sensitive and selective detection of Hg(2+). The results showed that there existed a linear relationship between the fluorescence intensity and the concentration of Hg(2+) from 0.001 to 1.0μM with a detection limit of 0.3nM. Finally, the sensor was successfully used to detection of Hg(2+) in water and milk samples.

  6. Highly sensitive and selective chip-based fluorescent sensor for mercuric ion: development and comparison of turn-on and turn-off systems.

    PubMed

    Du, Juan; Liu, Meiying; Lou, Xinhui; Zhao, Tao; Wang, Zheng; Xue, Ying; Zhao, Jianlong; Xu, Yuanshen

    2012-09-18

    Miniaturization is currently an important trend in environmental and food monitoring because it holds great promise for on-site monitoring and detection. We report here two ready-to-use chip-based fluorescent sensors, compatible with microarray technology for reagentless, one-step, fast, highly sensitive and selective detection of the mercuric ion (Hg(2+)) in the turn-on and turn-off operation modes. Both operation modes are based on the highly selective T-Hg(2+)-T coordination between two neighboring polythymine (T) strands at a high probe density and its induced displacement of the complementary polyadenine strand labeled with either fluorophore or quencher, which enables the turn-off and turn-on detection of Hg(2+), respectively. The turn-off sensor is slightly more sensitive than the turn-on sensor, and their detection limits are 3.6 and 8.6 nM, respectively, which are both lower than the U.S. Environmental Protection Agency limit of [Hg(2+)] for drinkable water (10 nM, 2 ppb). Compared to the turn-off sensor with the dynamic Hg(2+) detection range from 3.6 nM to 10 μM (R(2) = 0.99), the turn-on sensor has a broader dynamic Hg(2+) detection range, from 8.6 nM to 100 μM (R(2) = 0.996). Both sensors exhibited superior selectivity over other reported sensors using thymine-rich probes for Hg(2+) detection over other common metal ions. In addition, the practical application of the chip-based sensors was demonstrated by detecting spiked Hg(2+) in drinking water and fresh milk. The sensor has great potential for on-site practical applications due to its operational convenience, simplicity, speed, and portability.

  7. Visual test of subparts per billion-level mercuric ion with a gold nanoparticle probe after preconcentration by hollow fiber supported liquid membrane.

    PubMed

    Tan, Zhi-qiang; Liu, Jing-fu

    2010-05-15

    With the combination of the gold nanoparticle (AuNP)-based visual test with hollow fiber supported liquid membrane (HFSLM) extraction, a highly sensitive and selective method was developed for field detection of mercuric ion (Hg(2+)) in environmental waters. Hg(2+) in water samples was extracted through HFSLM and trapped in the aqueous acceptor and then visually detected based on the red-to-blue color change of 3-mercaptopropionic acid-functionalized AuNP (MPA-AuNP) probe. The highest extraction efficiency of Hg(2+) was obtained by using a 600 mL sample (pH 8.0, 2.0% (w/v) NaCl), approximately 35 microL of acceptor (10 mM of 2,6-pyridinedicarboxylic acid, pH 4.0) filled in the lumen of a polypropylene hollow fiber tubing (55 cm in length, 50 microm wall thickness, 280 microm inner diameter), a liquid membrane of 2.0% (w/v) trioctycphosphine oxide in undecane, and a shaking rate of 250 rpm. The chromegenic reaction was conducted by incubating the mixture of MPA-AuNP stock solution (12 microL, 15 nM), Tris-borate buffer solution (18 microL, 0.2 M, pH 9.5), and acceptor (30 microL) at 30 degrees C for 1 h. The detection limit can be adjusted to 0.8 microg/L Hg(2+) (corresponding to an enrichment factor of approximately 1000 in the HFSLM) and 2.0 microg/L Hg(2+) (the U.S. Environmental Protection Agency limit of [Hg(2+)] for drinkable water) by using extraction times of 3 and 1 h, respectively. The proposed method is extremely specific for Hg(2+) with tolerance to at least 1000-fold of other environmentally relevant heavy and transition metal ions and was successfully applied to detect Hg(2+) in a certified reference water sample, as well as real river, lake, and tap water samples.

  8. Chemically modified cellulose strips with pyridoxal conjugated red fluorescent gold nanoclusters for nanomolar detection of mercuric ions.

    PubMed

    Bothra, Shilpa; Upadhyay, Yachana; Kumar, Rajender; Ashok Kumar, S K; Sahoo, Suban K

    2017-04-15

    One-pot approach was adopted for the synthesis of highly luminescent near-infrared (NIR)-emitting gold nanoclusters (AuNCs) using bovine serum albumin (BSA) as a protecting agent. The vitamin B6 cofactor pyridoxal was conjugated with the luminescent BSA-AuNCs through the free amines of BSA and then employed for the nanomolar detection of Hg(2+) in aqueous medium via selective fluorescence quenching of AuNCs. This nano-assembly was successfully applied for the real sample analysis of Hg(2+) in fish, tap water and river water. The study also presents chemically-modified cellulosic paper strips with the pyridoxal conjugated BSA-AuNCs for detecting Hg(2+) ion up to 1nM.

  9. Mercuric ion attenuates nuclear factor-kappaB activation and DNA binding in normal rat kidney epithelial cells: implications for mercury-induced nephrotoxicity.

    PubMed

    Dieguez-Acuña, F J; Ellis, M E; Kushleika, J; Woods, J S

    2001-06-15

    Mercuric ion (Hg(2+)), one of the strongest thiol-binding agents known, mediates the toxicity associated with elemental, inorganic, and organic mercurial compounds. Studies of cellular events associated with Hg(2+) toxicity have focused largely on disruption of cell membranes and impairment of mitochondrial functions. In contrast, few studies have sought to define the specific molecular mechanisms through which Hg(2+) might affect toxicity via alteration of thiol-dependent signal transduction pathways that regulate cell proliferation and survival. Of particular interest in this regard is the effect of Hg(2+) on nuclear factor-kappaB (NF-kappaB), a pleiotropic transcriptional factor that is known to require reduced cysteine moieties at critical steps of activation and DNA binding. Here, we evaluated the effects of Hg(2+) on the expression of NF-kappaB in normal rat kidney epithelial (NRK52E) cells, a principal target of Hg(2+) toxicity. The lipopolysaccharide (LPS)-inducible form of NF-kappaB was readily detected in kidney cells and has been characterized as the p50p65 heterodimer. NF-kappaB-DNA binding was prevented in a dose-related manner by Hg(2+) (0-55 microM) in vitro when added to DNA binding reactions containing the nonthiol reducing agent Tris(2-carboxyethyl)phosphine hydrochloride (TCEP). Similarly, Hg(2+) at the same concentrations prevented DNA binding of a human recombinant wild-type p50p50 homodimer in binding reactions, and this effect was attenuated using a mutant form of the p50 protein containing a cys(62)-->ser(62) mutation. The inhibition of p50-DNA binding by Hg(2+) was reversible in a dose-related manner in vitro by competitive thiols DTT, GSH, and l-cysteine in binding reactions. In contrast, competitive thiols added to nuclear binding reactions were unable to reverse attenuation of LPS-mediated NF-kappaB-DNA binding affinity when cells were pretreated in vivo with Hg(2+) at concentrations as low as 2 microM prior to LPS administration

  10. Branchial and renal pathology in the fish exposed chronically to methoxy ethyl mercuric chloride

    SciTech Connect

    Gill, T.S.; Pant, J.C.; Tewari, H.

    1988-08-01

    Pathological manifestations causally related to pesticide poisoning have been described in both surficial and internal tissues of the fishes. Among the various organomercurials are phenyl mercuric acetate, methyl mercuric dicyanidiamide, methoxy ethyl mercuric chloride, methoxy ethyl mercuric silicate etc. Of these, the methoxy ethyl mercuric chloride (MEMC) is used in agriculture as an antifungal seed dressing, and its toxicity is primarily manifest in the Hg/sup 2 +/ ion. This report describes pathogenesis of branchial and renal lesions in the common freshwater fish, Puntius conchonius exposed chronically to sublethal levels of MEMC. Prior to this, alterations in the peripheral blood and metabolite levels in response to experimental MEMC poisoning have been demonstrated in this species.

  11. Thiol oxidase ability of copper ion is specifically retained upon chelation by aldose reductase.

    PubMed

    Balestri, Francesco; Moschini, Roberta; Cappiello, Mario; Mura, Umberto; Del-Corso, Antonella

    2017-06-01

    Bovine lens aldose reductase is susceptible to a copper-mediated oxidation, leading to the generation of a disulfide bridge with the concomitant incorporation of two equivalents of the metal and inactivation of the enzyme. The metal complexed by the protein remains redox active, being able to catalyse the oxidation of different physiological thiol compounds. The thiol oxidase activity displayed by the enzymatic form carrying one equivalent of copper ion (Cu1-AR) has been characterized. The efficacy of Cu1-AR in catalysing thiol oxidation is essentially comparable to the free copper in terms of both thiol concentration and pH effect. On the contrary, the two catalysts are differently affected by temperature. The specificity of the AR-bound copper towards thiols is highlighted with Cu1-AR being completely ineffective in promoting the oxidation of both low-density lipoprotein and ascorbic acid.

  12. Neutron Detection with Mercuric Iodide

    SciTech Connect

    Bell, Z.A.

    2003-06-17

    Mercuric iodide is a high-density, high-Z semiconducting material useful for gamma ray detection. This makes it convertible to a thermal neutron detector by covering it with a boron rich material and detecting the 478 keV gamma rays resulting from the {sup 10}B(n, {alpha}){sup 7}Li* reaction. However, the 374 barn thermal capture cross section of {sup nat}Hg, makes the detector itself an attractive absorber, and this has been exploited previously. Since previous work indicates that there are no low-energy gamma rays emitted in coincidence with the 368 keV capture gamma from the dominant {sup 199}Hg(n, {gamma}){sup 200}Hg reaction, only the 368 keV capture gamma is seen with any efficiency a relatively thin (few mm) detector. In this paper we report preliminary measurements of neutrons via capture reactions in a bare mercuric iodide crystal and a crystal covered in {sup 10}B-loaded epoxy. The covered detector is an improvement over the bare detector because the presence of both the 478 and 368 keV gamma rays removes the ambiguity associated with the observation of only one of them. Pulse height spectra, obtained with and without lead and cadmium absorbers, showed the expected gamma rays and demonstrated that they were caused by neutrons.

  13. Mercuric iodide light detector and related method

    DOEpatents

    Iwanczyk, Jan S.; Barton, Jeff B.; Dabrowski, Andrzej J.; Schnepple, Wayne F.

    1986-01-01

    Apparatus and method for detecting light involve applying a substantially uniform electrical potential difference between first and second spaced surfaces of a body of mercuric iodide, exposing the first surface to light and measuring an electrical current passed through the body in response to the light. The mercuric iodide may be substantially monocrystalline and the potential may be applied between a substantially transparent conductive layer at the first surface and a second conductive layer at the second surface. In a preferred embodiment, the detector is coupled to a scintillator for passage of light to the mercuric iodide in response to ionizing radiation incident on the scintillator.

  14. Large area mercuric iodide photodetectors

    SciTech Connect

    Iwanczyk, J.S.; Dabrowski, A.J.; Markakis, J.M.; Ortale, C.; Schnepple, W.F.

    1984-02-01

    Results of an investigation of large area mercuric iodide (HgI/sub 2/) photodetectors are reported. Different entrance contacts were studied, including semitransparent metallic films and conductive liquids. Theoretical calculations of electronic noise of these photodetectors were compared with experimental results. HgI/sub 2/ photodetectors with active area up to 4 cm/sup 2/ were matched with NaI(Tl) and CsI(Tl) scintillation crystals and were evaluated as gamma-radiation spectrometers. Energy resolution of 9.3% for gamma radiation of 511 keV with a CsI(Tl) scintillator and energy resolution of 9.0% for gamma radiation of 622 keV with a NaI(Tl) scintillator have been obtained.

  15. A novel optical DNA biosensor for detection of trace amounts of mercuric ions using gold nanoparticles introduced onto modified glass surface

    NASA Astrophysics Data System (ADS)

    Mashhadizadeh, Mohammad Hossein; Talemi, Rasoul Pourtaghavi

    2014-11-01

    In this work we report a DNA spectrophotometric biosensor for detection of Hg2+ ions in which a pair of oligonucleotides with four thymine-thymine (T-T) mismatched bases was immobilized onto modified glass surface. Firstly, glass surface modified with 3-(mercaptopropyl) trimethoxysilane (MSPT) and gold nano-particles respectively and then one oligonucleotide (P1) modified with hexanthiol at 5-terminal was immobilized on gold nano-particles via self-assembly and inserted in methylene blue. Methylene blue can intercalate on single strand DNA (ss-DNA) and its absorption peak can measure spectrophotometrically. Then the other oligonucleotide was able to hybridize with P1 by forming thymine-Hg2+-thymine (T-Hg2+-T) complexes in the presence of Hg2+, and absorption signal of methylene blue reduced upon Hg2+ increasing concentration because inaccessibility of guanine base in DNA duplex. However, when Hg2+ was absent, the two oligonucleotides could not hybridize due to the T-T mismatched bases, and P2 could not be fixed on the modified glass surface and any change in absorption peak of methylene blue takes place. The UV-Vis spectrum showed a linear correlation between the absorption peak of methylene blue and the concentration of Hg2+ over the range from 10 nM to 10 μM (R2 = 0.9985) with a detection limit of 6 nM. This spectrophotometric biosensor could be widely used for selective detection of Hg2+.

  16. The orphan protein bis-γ-glutamylcystine reductase joins the pyridine nucleotide-disulfide reductase family

    PubMed Central

    Kim, Juhan; Copley, Shelley D.

    2014-01-01

    Facile DNA sequencing became possible decades after many enzymes had been purified and characterized. Consequently, there are still “orphan” enyzmes whose activity is known but the genes that encode them have not been identified. Identification of the genes encoding orphan enzymes is important because it allows correct annotation of genes of unknown function or with mis-assigned function. Bis-γ-glutamylcystine reductase (GCR) is an orphan protein that was purified in 1988. This enzyme catalyzes the reduction of bis-γ-glutamylcystine. γ-Glutamylcysteine (γ-Glu-Cys) is the major low molecular weight thiol in halobacteria. We purified GCR from Halobacterium sp. NRC-1 and identified the sequence of 23 tryptic peptides by NanoLC electrospray ionization tandem mass spectrometry. These peptides cover 62% of the protein predicted to be encoded by a gene in Halobacterium sp. NRC-1 that is annotated as mercuric reductase. GCR and mercuric reductase activities were assayed using enzyme that was expressed in E. coli and re-folded from inclusion bodies. The enzyme had robust GCR activity, but no mercuric reductase activity. The genomes of most, but not all, halobacteria for which whole genome sequences are available have close homologs of GCR, suggesting that there is more to be learned about the low molecular weight thiols used in halobacteria. PMID:23560638

  17. Thermal decomposition of mercuric sulfide

    SciTech Connect

    Leckey, J.H.; Nulf, L.E.

    1994-10-28

    The rate of thermal decomposition of mercuric sulfide (HgS) has been measured at temperatures from 265 to 345 C. These data have been analyzed using a first-order chemical reaction model for the time dependence of the reaction and the Arrhenius equation for the temperature dependence of the rate constant. Using this information, the activation energy for the reaction was found to be 55 kcal/mol. Significant reaction vessel surface effects obscured the functional form of the time dependence of the initial portion of the reaction. The data and the resulting time-temperature reaction-rate model were used to predict the decomposition rate of HgS as a function of time and temperature in thermal treatment systems. Data from large-scale thermal treatment studies already completed were interpreted in terms of the results of this study. While the data from the large-scale thermal treatment studies were consistent with the data from this report, mass transport effects may have contributed to the residual amount of mercury which remained in the soil after most of the large-scale runs.

  18. Energy resolution enhancement of mercuric iodide detectors

    NASA Technical Reports Server (NTRS)

    Finger, M.; Prince, T. A.; Padgett, L.; Prickett, B.; Schnepple, W.

    1984-01-01

    A pulse processing technique has been developed which improves the gamma-ray energy resolution of mercuric iodide detectors. The technique employs a fast (100 ns) and a slow (6.4 microsec) pulse height analysis to correct for signal variations due to variations in charge trapping. The capabilities of the technique for energy resolution enhancement are discussed as well as the utility of the technique for examining the trapping characteristics of individual detectors. An energy resolution of 2.6 percent FWHM at 662 keV was achieved with an acceptance efficiency of 100 percent from a mercuric iodide detector which gives 8.3 percent FWHM using standard techniques.

  19. Inhibitory effects of calmodulin antagonists on urinary enzyme excretion in rats after nephrotoxic doses of mercuric chloride

    SciTech Connect

    Harrison, S.D. Jr.; Cox, J.L.; Giles, R.C. Jr.

    1985-03-01

    Prochlorperazine, a phenothiazine antiemetic, has been reported to protect rats against mercuric chloride (HgCl/sub 2/)-induced nephrotoxicity. Mercuric ion and 12 other divalent metal ions of toxicologic importance inhibit the activity of calmodulin, a ubiquitous intracellular calcium receptor and regulatory protein, at physiologically relevant concentrations. Phenothiazines, including prochlorperazine, are reversible calmodulin antagonists, and as such they interact with divalent calcium at the level of calmodulin. It was of interest therefore to evaluate the comparative effects of several phenothiazines on HgCl/sub 2/-induced nephrotoxicity in rats.

  20. Roles of glutamates and metal ions in a rationally designed nitric oxide reductase based on myoglobin

    SciTech Connect

    Lin, Y.W.; Robinson, H.; Yeung, N.; Gao, Y.-G.; Miner, K. D.; Tian, S.; Lu, Y.

    2010-05-11

    A structural and functional model of bacterial nitric oxide reductase (NOR) has been designed by introducing two glutamates (Glu) and three histidines (His) in sperm whale myoglobin. X-ray structural data indicate that the three His and one Glu (V68E) residues bind iron, mimicking the putative FeB site in NOR, while the second Glu (I107E) interacts with a water molecule and forms a hydrogen bonding network in the designed protein. Unlike the first Glu (V68E), which lowered the heme reduction potential by {approx}110 mV, the second Glu has little effect on the heme potential, suggesting that the negatively charged Glu has a different role in redox tuning. More importantly, introducing the second Glu resulted in a {approx}100% increase in NOR activity, suggesting the importance of a hydrogen bonding network in facilitating proton delivery during NOR reactivity. In addition, EPR and X-ray structural studies indicate that the designed protein binds iron, copper, or zinc in the FeB site, each with different effects on the structures and NOR activities, suggesting that both redox activity and an intermediate five-coordinate heme-NO species are important for high NOR activity. The designed protein offers an excellent model for NOR and demonstrates the power of using designed proteins as a simpler and more well-defined system to address important chemical and biological issues.

  1. Roles of Glutamates and Metal ions in a Rationally Designed Nitric Oxide Reductase Based on Myoglobin

    SciTech Connect

    Y Lin; N Yeung; Y Gao; K Miner; S Tian; H Robinson; Y Lu

    2011-12-31

    A structural and functional model of bacterial nitric oxide reductase (NOR) has been designed by introducing two glutamates (Glu) and three histidines (His) in sperm whale myoglobin. X-ray structural data indicate that the three His and one Glu (V68E) residues bind iron, mimicking the putative FeB site in NOR, while the second Glu (I107E) interacts with a water molecule and forms a hydrogen bonding network in the designed protein. Unlike the first Glu (V68E), which lowered the heme reduction potential by {approx}110 mV, the second Glu has little effect on the heme potential, suggesting that the negatively charged Glu has a different role in redox tuning. More importantly, introducing the second Glu resulted in a {approx}100% increase in NOR activity, suggesting the importance of a hydrogen bonding network in facilitating proton delivery during NOR reactivity. In addition, EPR and X-ray structural studies indicate that the designed protein binds iron, copper, or zinc in the FeB site, each with different effects on the structures and NOR activities, suggesting that both redox activity and an intermediate five-coordinate heme-NO species are important for high NOR activity. The designed protein offers an excellent model for NOR and demonstrates the power of using designed proteins as a simpler and more well-defined system to address important chemical and biological issues.

  2. Three spinach leaf nitrate reductase-3-hydroxy-3-methylglutaryl-CoA reductase kinases that are required by reversible phosphorylation and/or Ca2+ ions.

    PubMed Central

    Douglas, P; Pigaglio, E; Ferrer, A; Halfords, N G; MacKintosh, C

    1997-01-01

    In spinach (Spinacea oleracea L.) leaf extracts, three protein kinases (PKI, PKII and PKIII) were identified each of which phosphorylated spinach nitrate reductase on serine-543, and inactivated the enzyme in the presence of nitrate reductase inhibitor, 14-3-3. PKIII was also very active in phosphorylating and inactivating Arabidopsis (Landsberg erecta) 3-hydroxy-3-methylglutaryl-coenzyme A reductase 1 (HMGR1). PKI and PKII phosphorylated HMGR1 more slowly than PKIII, compared with their relative rates of phosphorylation of nitrate reductase. HMGR1 identical with those that are seen after phosphorylation of serine-577 by the sucrose non-fermenting (SNF1)-like PK, 3-hydroxy-3-methylglutaryl-Co A reductase kinase A (HRK-A), from cauliflower [Dale, Arró, Becerra, Morrice, Boronat, Hardie and Ferrer (1995) Eur. J. Biochem. 233, 506-513]. PKI was Ca2+-dependent when prepared in the absence of protein phosphatase (PP) inhibitors, and largely Ca2+-dependent when prepared in the presence of PP inhibitors (NaF and EGTA). The Ca2+-independent portion of PKI was inactivated by either PP2A or PP2C, while the Ca2+-dependent portion of PKI became increasingly activated during storage, which we presume was mimicking the effect of an unidentified PP. These findings indicate that PK1 is regulated by two functionally distinct phosphorylations. PKI had a molecular mass of 45 kDa on gel filtration and was active towards substrate peptides that terminated at the +2 residue from the phosphorylation site, whereas PKIII was inactive towards these peptides. PKII was Ca2+-stimulated under all conditions tested. PKIII was Ca2+-indepdented, inactivated by PP2A or PP2C, had a requirement for a hydrophobic residue in the +4 position of peptide substrates, had a molecular mass by gel filtration of approximately 140 kDa, and an antibody against the rye SNF1-related PK (RKIN1) recognized a 58 kDa subunit in fractions containing PKIII. These properties of PKIII are identical with those reported

  3. Mercuric chloride (HgCl2)

    Integrated Risk Information System (IRIS)

    Mercuric chloride ( HgCl2 ) ; CASRN 7487 - 94 - 7 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Nonc

  4. MERCURIC CHLORIDE CAPTURE BY ALKALINE SORBENTS

    EPA Science Inventory

    The paper gives results of bench-scale mechanistic studies of mercury/sorbent reactions that showed that mercuric chloride (HgC12) is readily adsorbed by alkaline sorbents, which may offers a less expensive alternative to the use of activated carbons. A laboratory-scale, fixed-b...

  5. MERCURIC CHLORIDE CAPTURE BY ALKALINE SORBENTS

    EPA Science Inventory

    The paper gives results of bench-scale mechanistic studies of mercury/sorbent reactions that showed that mercuric chloride (HgC12) is readily adsorbed by alkaline sorbents, which may offers a less expensive alternative to the use of activated carbons. A laboratory-scale, fixed-b...

  6. Mercuric iodide X-ray camera

    NASA Astrophysics Data System (ADS)

    Patt, B. E.; del Duca, A.; Dolin, R.; Ortale, C.

    1986-02-01

    A prototype X-ray camera utilizing a 1.5- by 1.5-in., 1024-element, thin mercuric iodide detector array has been tested and evaluated. The microprocessor-based camera is portable and operates at room temperature. Events can be localized within 1-2 mm at energies below 60 keV and within 5-6 mm at energies on the order of 600 keV.

  7. Determination of mercurous chloride and total mercury in mercury ores

    USGS Publications Warehouse

    Fahey, J.J.

    1937-01-01

    A method for the determination of mercurous chloride and total mercury on the same sample is described. The mercury minerals are volatilized in a glass tube and brought into intimate contact with granulated sodium carbonate. The chlorine is fixed as sodium chloride, determined with silver nitrate, and computed to mercurous chloride. The mercury is collected on a previously weighed gold coil and weighed.

  8. 34. August, 1971. PHOTOCOPY: GENERAL VIEW OF CITY OF MERCUR ...

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

    34. August, 1971. PHOTOCOPY: GENERAL VIEW OF CITY OF MERCUR CA. 1910 (THIS HISTORIC VIEW IS TAKEN FROM A PUBLICATION BY UTAH POWER & LIGHT CO. CREDIT REQUESTED TO COMPANY.). (SEE UT-10-2 FOR PRESENT DAY VIEW). - DeLamar Mercur Mines Company, Golden Gate Mill, Ophir, Tooele County, UT

  9. Arsenate reductase from Thermus thermophilus conjugated to polyethylene glycol-stabilized gold nanospheres allow trace sensing and speciation of arsenic ions

    PubMed Central

    Politi, Jane; Spadavecchia, Jolanda; Fiorentino, Gabriella; Antonucci, Immacolata

    2016-01-01

    Water sources pollution by arsenic ions is a serious environmental problem all around the world. Arsenate reductase enzyme (TtArsC) from Thermus thermophilus extremophile bacterium, naturally binds arsenic ions, As(V) and As (III), in aqueous solutions. In this research, TtArsC enzyme adsorption onto hybrid polyethylene glycol-stabilized gold nanoparticles (AuNPs) was studied at different pH values as an innovative nanobiosystem for metal concentration monitoring. Characterizations were performed by UV/Vis and circular dichroism spectroscopies, TEM images and in terms of surface charge changes. The molecular interaction between arsenic ions and the TtArsC-AuNPs nanobiosystem was also monitored at all pH values considered by UV/Vis spectroscopy. Tests performed revealed high sensitivities and limits of detection equal to 10 ± 3 M−12 and 7.7 ± 0.3 M−12 for As(III) and As(V), respectively. PMID:27707908

  10. Synthesis of α-Diketones from Alkylaryl- and Diarylalkynes Using Mercuric Salts

    PubMed Central

    2015-01-01

    Both alkylarylalkynes and diarylalkynes 1 are converted into the α-diketones 2 in good yield by the use of mercuric salts, e.g., mercuric nitrate hydrate or mercuric triflate, in the presence of water. Other mercuric salts, e.g., sulfate, chloride, acetate, or trifluoroacetate, do not provide the diketone product. A possible mechanism is proposed. PMID:24684513

  11. Synthetic and structural investigations of mercurous and mercuric organophosphonates and phenylarsonates

    NASA Astrophysics Data System (ADS)

    Padalwar, Nitin Balkrushna; Vidyasagar, Kanamaluru

    2016-11-01

    The following twelve mercurous and mercuric organophosphomates, bis/diphosphonates and phenylarsonates have been isolated and structurally characterized by single crystal X-ray diffraction, 13C-and 31P NMR, infrared and Raman spectroscopic methods: Hg2(HO3PC6H5)2(1), Hg2(HO3P(C6H4)PO3H)(2), Hg2(HO3P(C6H4)2PO3H)(3), Hg2(HO3P(CH2)4PO3H)(4), Hg2(O3PC6H5)·H2O(5), (Hg2)2(O3P(CH2)2PO3)(6), (Hg2)2(O3P(CH2)3PO3)(7), Hg(O3PC6H5)·H2O(8), Hg(O3PCH2C6H5)·H2O(9), Hg(O3AsC6H5)·H2O(10), Hg3(O3AsC6H5)2(HO3AsC6H5)2(11) and (Hg2)Hg3(O3P(C6H4)PO3)2·2H2O(12). Compounds 1-7 are the first examples of mercurous phosphonates and di/bisphosphonates. They contain Hg2O6 units, which consist of Hg22+ cations with Hg-Hg bond of 2.5 Å length. Phenylphosphonates 1 and 5 are layered compounds, whereas bis/diphosphonates 2, 3, 4, 6 and 7 have pillared-layered and three-dimensional structures. Compounds 8-11 are layered mercuric phosphonates and phenylarsonates. Compound 12 is a three-dimensional mixed-valent mercury phenylenebisphosphonate.

  12. Heavy metal resistance in Arthrobacter ramosus strain G2 isolated from mercuric salt-contaminated soil.

    PubMed

    Bafana, Amit; Krishnamurthi, Kannan; Patil, Mahendra; Chakrabarti, Tapan

    2010-05-15

    Present study describes isolation of a multiple metal-resistant Arthrobacter ramosus strain from mercuric salt-contaminated soil. The isolate was found to resist and bioaccumulate several metals, such as cadmium, cobalt, zinc, chromium and mercury. Maximum tolerated concentrations for above metals were found to be 37, 525, 348, 1530 and 369 microM, respectively. The isolate could also reduce and detoxify redox-active metals like chromium and mercury, indicating that it has great potential in bioremediation of heavy metal-contaminated sites. Chromate reductase and mercuric reductase (MerA) activities in protein extract of the culture were found to be 2.3 and 0.17 units mg(-1) protein, respectively. MerA enzyme was isolated from the culture by (NH(4))(2)SO(4) precipitation followed by dye affinity chromatography and its identity was confirmed by nano-LC-MS/MS. Its monomeric molecular weight, and optimum pH and temperature were 57kDa, 7.4 and 55 degrees C, respectively. Thus, the enzyme was mildly thermophilic as compared to other MerA enzymes. K(m) and V(max) of the enzyme were 16.9 microM HgCl(2) and 6.2 micromol min(-1)mg(-1) enzyme, respectively. The enzyme was found to be NADPH-specific. To our knowledge this is the first report on characterization of MerA enzyme from an Arthrobacter sp. Copyright (c) 2009 Elsevier B.V. All rights reserved.

  13. Mercuric iodine room temperature gamma-ray detectors

    NASA Technical Reports Server (NTRS)

    Patt, Bradley E.; Markakis, Jeffrey M.; Gerrish, Vernon M.; Haymes, Robert C.; Trombka, Jacob I.

    1990-01-01

    high resolution mercuric iodide room temperature gamma-ray detectors have excellent potential as an essential component of space instruments to be used for high energy astrophysics. Mercuric iodide detectors are being developed both as photodetectors used in combination with scintillation crystals to detect gamma-rays, and as direct gamma-ray detectors. These detectors are highly radiation damage resistant. The list of applications includes gamma-ray burst detection, gamma-ray line astronomy, solar flare studies, and elemental analysis.

  14. Mercuric iodine room temperature gamma-ray detectors

    NASA Technical Reports Server (NTRS)

    Patt, Bradley E.; Markakis, Jeffrey M.; Gerrish, Vernon M.; Haymes, Robert C.; Trombka, Jacob I.

    1990-01-01

    high resolution mercuric iodide room temperature gamma-ray detectors have excellent potential as an essential component of space instruments to be used for high energy astrophysics. Mercuric iodide detectors are being developed both as photodetectors used in combination with scintillation crystals to detect gamma-rays, and as direct gamma-ray detectors. These detectors are highly radiation damage resistant. The list of applications includes gamma-ray burst detection, gamma-ray line astronomy, solar flare studies, and elemental analysis.

  15. A new strategy for strain improvement of Aurantiochytrium sp. based on heavy-ions mutagenesis and synergistic effects of cold stress and inhibitors of enoyl-ACP reductase.

    PubMed

    Cheng, Yu-Rong; Sun, Zhi-Jie; Cui, Gu-Zhen; Song, Xiaojin; Cui, Qiu

    2016-11-01

    Developing a strain with high docosahexaenoic acid (DHA) yield and stable fermenting-performance is an imperative way to improve DHA production using Aurantiochytrium sp., a microorganism with two fatty acid synthesis pathways: polyketide synthase (PKS) pathway and Type I fatty acid synthase (FAS) pathway. This study investigated the growth and metabolism response of Aurantiochytrium sp. CGMCC 6208 to two inhibitors of enoyl-ACP reductase of Type II FAS pathway (isoniazid and triclosan), and proposed a method of screening high DHA yield Aurantiochytrium sp. strains with heavy ion mutagenesis and pre-selection by synergistic usage of cold stress (4°C) and FAS inhibitors (triclosan and isoniazid). Results showed that (1) isoniazid and triclosan have positive effects on improving DHA level of cells; (2) mutants from irradiation dosage of 120Gy yielded more DHA compared with cells from 40Gy, 80Gy treatment and wild type; (3) DHA contents of mutants pre-selected by inhibitors of enoyl-ACP reductase of Type II FAS pathway (isoniazid and triclosan)at 4°C, were significantly higher than that of wild type; (4) compared to the wild type, the DHA productivity and yield of a mutant (T-99) obtained from Aurantiochytrium sp. CGMCC 6208 by the proposed method increased by 50% from 0.18 to 0.27g/Lh and 30% from 21 to 27g/L, respectively. In conclusion, this study developed a feasible method to screen Aurantiochytrium sp. with high DHA yield by a combination of heavy-ion mutagenesis and mutant-preselection by FAS inhibitors and cold stress. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Near-infrared to near-infrared upconverting NaYF4:Yb3+,Tm3+ nanoparticles-aptamer-Au nanorods light resonance energy transfer system for the detection of mercuric(II) ions in solution.

    PubMed

    Chen, Hong-Qi; Yuan, Fei; Wang, Shao-Zhen; Xu, Juan; Zhang, Yi-Yan; Wang, Lun

    2013-04-21

    A new luminescence resonant energy transfer (LRET) system has been designed that utilizes near-infrared (NIR)-to-NIR upconversion lanthanide nanophosphors (UCNPs) as the donor, and Au nanorods (Au NRs) as the acceptor. The UCNPs were excited by a near-infrared (980 nm) wavelength and also emitted at a near-infrared wavelength (804 nm) using an inexpensive infrared continuous wave laser diode. The Au NRs showed a high absorption band around 806 nm, which provided large spectral overlap between the donor and the acceptor. Hg(2+) ions were added to an aqueous solution containing the UCNPs and Au NRs that were modified with a Hg(2+) aptamer. Then, a sandwich-type LRET system was developed for the detection of Hg(2+) ions that had high sensitivity and selectivity in the NIR region. The method was successfully applied to the sensing of Hg(2+) ions in water and human serum samples.

  17. Arsenate reductase from Thermus thermophilus conjugated to polyethylene glycol-stabilized gold nanospheres allow trace sensing and speciation of arsenic ions.

    PubMed

    Politi, Jane; Spadavecchia, Jolanda; Fiorentino, Gabriella; Antonucci, Immacolata; De Stefano, Luca

    2016-10-01

    Water sources pollution by arsenic ions is a serious environmental problem all around the world. Arsenate reductase enzyme (TtArsC) from Thermus thermophilus extremophile bacterium, naturally binds arsenic ions, As(V) and As (III), in aqueous solutions. In this research, TtArsC enzyme adsorption onto hybrid polyethylene glycol-stabilized gold nanoparticles (AuNPs) was studied at different pH values as an innovative nanobiosystem for metal concentration monitoring. Characterizations were performed by UV/Vis and circular dichroism spectroscopies, TEM images and in terms of surface charge changes. The molecular interaction between arsenic ions and the TtArsC-AuNPs nanobiosystem was also monitored at all pH values considered by UV/Vis spectroscopy. Tests performed revealed high sensitivities and limits of detection equal to 10 ± 3 M(-12) and 7.7 ± 0.3 M(-12) for As(III) and As(V), respectively. © 2016 The Author(s).

  18. Mercuric iodide photodetectors for scintillation spectroscopy

    SciTech Connect

    Markakis, J.; Dabrowski, A.; Iwanczyk, J.; Ortale, C.; Schnepple, W.

    1985-02-01

    We have measured the responses to /sup 137/Cs (662 keV) of both a 1-inch-diam by 2-inch-thick NaI(Tl) scintillator optically coupled to a 1-inch-diam by 800-..mu..mthick mercuric iodide (HgI/sub 2/) photodetector, and a 1-cmdiam by 1-cm-thick CaWO/sub 4/ scintillator coupled to a 1.3-cm-diam by 600-..mu..m-thick HgI/sub 2/ photodetector. Best spectral resolution to /sup 137/Cs was 7.8% FWHM for the NaI(Tl)-HgI/sub 2/ and 12.5% FWHM for the CaWO/sub 4/-HgI/sub 2/ detectors; peak-to-valley ratios were 26:1 and 16:1, respectively. HgI/sub 2/ detectors operate at room temperature and their use in scintillation spectroscopy presents the ultimate miniaturization of scintillation detectors, limited mainly by the size of the scintillation crystal.

  19. Mercuric iodide photodetectors for scintillation spectroscopy

    SciTech Connect

    Markakis, J.; Ortale, C.; Schnepple, W.; Iwanczyk, J.; Dabrowski, A.

    1984-01-01

    We have measured the responses to /sup 137/Cs (662 keV) of both a 1-inch-diam by 2-inch-thick NaI(Tl) scintillator optically coupled to a 1-inch-diam by 800-..mu..m-thick mercuric iodide (HgI/sub 2/) photodetector, and a 1-cm-diam by 1-cm-thick CaWO/sub 4/ scintillator coupled to a 1.3-cm-diam by 600-..mu..m-thick HgI/sub 2/ photodetector. Best spectral resolution to /sup 137/Cs was 7.8% FWHM for the NaI(Tl)-HgI/sub 2/ and 12.5% FWHM for the CaWO/sub 4/-HgI/sub 2/ detectors; peak-to-valley ratios were 26:1 and 16:1, respectively. HgI/sub 2/ detectors operate at room temperature and their use in scintillation spectroscopy presents the ultimate miniaturization of scintillation detectors, limited mainly by the size of the scintillation crystal.

  20. Defects and impurities in mercuric iodine processing

    SciTech Connect

    van Scyoc, J.M.; James, R.B.; Schlesinger, T.E.; Gilbert, T.S.

    1996-03-01

    In the fabrication of mercuric iodide HgI{sub 2} room temperature radiation detectors, as in any semiconductor process, the quality of the final device is very sensitive to the impurities and defects present. Each process step can change the effects of existing defects, reduce the number of defects, or introduce new defects. In HgI{sub 2} detectors these defects act as trapping and recombination centers, thereby degrading immediate performance and leading to unstable devices. In this work we characterized some of the defects believed to strongly affect detector operation. Specifically, we studied impurities that are known to be present in typical HgI{sub 2} materials. Leakage current measurements were used to study the introduction and characteristics of these impurities, as such experiments reveal the mobile nature of these defects. In particular, we found that copper, which acts as a hole trap, introduces a positively charged center that diffuses and drifts readily in typical device environments. These measurements suggest that Cu, and related impurities like silver, may be one of the leading causes of HgI{sub 2} detector failures.

  1. Development of a mercuric iodide solid state spectrometer for X-ray astronomy

    NASA Technical Reports Server (NTRS)

    Vallerga, J.

    1983-01-01

    Mercuric iodide detectors, experimental development for astronomical use, X ray observations of the 1980 Cygnus X-1 High State, astronomical had X ray detectors in current use, detector development, balloon flight of large area (1500 sq cm) Phoswich detectors, had X ray telescope design, shielded mercuric iodide background measurement, Monte Carlo analysis, measurements with a shielded mercuric iodide detector are discussed.

  2. Nitrate reductase mutation alters potassium nutrition as well as nitric oxide-mediated control of guard cell ion channels in Arabidopsis.

    PubMed

    Chen, Zhong-Hua; Wang, Yizhou; Wang, Jian-Wen; Babla, Mohammad; Zhao, Chenchen; García-Mata, Carlos; Sani, Emanuela; Differ, Christopher; Mak, Michelle; Hills, Adrian; Amtmann, Anna; Blatt, Michael R

    2016-03-01

    Maintaining potassium (K(+) ) nutrition and a robust guard cell K(+) inward channel activity is considered critical for plants' adaptation to fluctuating and challenging growth environment. ABA induces stomatal closure through hydrogen peroxide and nitric oxide (NO) along with subsequent ion channel-mediated loss of K(+) and anions. However, the interactions of NO synthesis and signalling with K(+) nutrition and guard cell K(+) channel activities have not been fully explored in Arabidopsis. Physiological and molecular techniques were employed to dissect the interaction of nitrogen and potassium nutrition in regulating stomatal opening, CO2 assimilation and ion channel activity. These data, gene expression and ABA signalling transduction were compared in wild-type Columbia-0 (Col-0) and the nitrate reductase mutant nia1nia2. Growth and K(+) nutrition were impaired along with stomatal behaviour, membrane transport, and expression of genes associated with ABA signalling in the nia1nia2 mutant. ABA-inhibited K(+) in current and ABA-enhanced slow anion current were absent in nia1nia2. Exogenous NO restored regulation of these channels for complete stomatal closure in nia1nia2. While NO is an important signalling component in ABA-induced stomatal closure in Arabidopsis, our findings demonstrate a more complex interaction associating potassium nutrition and nitrogen metabolism in the nia1nia2 mutant that affects stomatal function. © 2015 The Authors. New Phytologist © 2015 New Phytologist Trust.

  3. Introduction of extrinsic defects into mercuric iodide during processing

    SciTech Connect

    Hung, C.; Bao, X.J.; Schlesinger, T.E. ); James, R.B. ); Cheng, A.Y.; Ortale, C.; van den Berg, L. )

    1993-05-01

    Low temperature (4.2 K) photoluminescence spectroscopy (PL) measurements were performed on mercuric iodide (HgI[sub 2]) crystals which were intentionally doped with copper or silver during KI etching. PL spectra obtained after these doping experiments show specific Cu and Ag features similar to those previously observed after deposition of Cu or Ag contacts on mercuric iodide crystals. The in-diffusion of Cu or Ag into bulk HgI[sub 2] has also been confirmed a few days after doping. This diffusion introduces new recombination centers in the material. This work suggests that the processing steps used to fabricate mercuric iodide nuclear detectors can lead to the introduction of new defects which are detrimental to detector performance.

  4. Thermodynamic reactivity, growth and characterization of mercurous halide crystals

    NASA Technical Reports Server (NTRS)

    Singh, N. B.; Gottlieb, M.; Henningsen, T.; Hopkins, R. H.; Mazelsky, R.; Singh, M.; Glicksman, M. E.; Paradies, C.

    1992-01-01

    Thermodynamic calculations were carried out for the Hg-X-O system (X = Cl, Br, I) to identify the potential sources of contamination and relative stability of oxides and oxy-halide phases. The effect of excess mercury vapor pressure on the optical quality of mercurous halide crystal was studied by growing several mercurous chloride crystals from mercury-rich composition. The optical quality of crystals was examined by birefringence interferometry and laser scattering studies. Crystals grown in slightly mercury-rich composition showed improved optical quality relative to stoichiometric crystals.

  5. Chronic effects of mercuric chloride ingestion on rat adrenocortical function

    SciTech Connect

    Agrawal, R.; Chansouria, J.P.N. )

    1989-09-01

    Mercurial contamination of environment has increased. Mercury accumulates in various organs and adversely affects their functions. Some of the most prominent toxic effects of inorganic mercury compounds include neurotoxicity, hepatotoxicity and nephrotoxicity. Besides this, mercury has also been reported to affect various endocrine glands like pituitary, thyroid, gonadal and adrenal glands. There have been no reports on the toxic effects of chronic oral administration of varying doses of mercuric chloride on adrenocortical function in albino rats. The present work was undertaken to study the adrenocortical response to chronic oral administration of mercuric chloride of varying dose and duration in albino rats.

  6. The Role of Nitrate and Ammonium Ions and Light on the Induction of Nitrate Reductase in Maize Leaves 1

    PubMed Central

    Oaks, Ann; Poulle, Michel; Goodfellow, Valerie J.; Cass, Leslie A.; Deising, Holger

    1988-01-01

    Corn seedlings (Zea mays cv W64A × W182E) were grown hydroponically, in the presence or absence of NO3−, with or without light and with NH4Cl as the only N source. In agreement with earlier results nitrate reductase (NR) activity was found only in plants treated with both light and NO3−. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by transfer of the proteins to nitrocellulose paper and reaction with antibodies prepared against a pure NR showed that crude extracts prepared from light-grown plants had a polypeptide of approximately 116 kilodaltons (the subunit size for NR) when NO3− was present in the growth medium. Crude extracts from plants grown in the dark did not have the 116 kilodalton polypeptide, although smaller polypeptides, which reacted with NR-immunoglobulin G, were sometimes found at the gel front. When seedlings were grown on Kimpack paper or well washed sand, NR activity was again found only when the seedlings were exposed to light and NO3−. Under these conditions, however, a protein of about 116 kilodaltons, which reacted with the NR antibody was present in light-grown plants whether NO3− was added to the system or not. The NR antibody cross-reacting protein was also seen in hydroponically grown plants when NH4Cl− was the only added form of nitrogen. These results indicate that the induction of an inactive NR-protein precursor in corn is mediated either by extremely low levels of NO3− or by some other unidentified factor, and that higher levels of NO3− are necessary for converting the inactive NR cross-reacting protein to a form of the enzyme capable of reducing NO3− to NO2−. Images Fig. 1 Fig. 2 Fig. 3 PMID:16666423

  7. Thioredoxin reductase.

    PubMed Central

    Mustacich, D; Powis, G

    2000-01-01

    The mammalian thioredoxin reductases (TrxRs) are a family of selenium-containing pyridine nucleotide-disulphide oxidoreductases with mechanistic and sequence identity, including a conserved -Cys-Val-Asn-Val-Gly-Cys- redox catalytic site, to glutathione reductases. TrxRs catalyse the NADPH-dependent reduction of the redox protein thioredoxin (Trx), as well as of other endogenous and exogenous compounds. The broad substrate specificity of mammalian TrxRs is due to a second redox-active site, a C-terminal -Cys-SeCys- (where SeCys is selenocysteine), that is not found in glutathione reductase or Escherichia coli TrxR. There are currently two confirmed forms of mammalian TrxRs, TrxR1 and TrxR2, and it is possible that other forms will be identified. The availability of Se is a key factor determining TrxR activity both in cell culture and in vivo, and the mechanism(s) for the incorporation of Se into TrxRs, as well as the regulation of TrxR activity, have only recently begun to be investigated. The importance of Trx to many aspects of cell function make it likely that TrxRs also play a role in protection against oxidant injury, cell growth and transformation, and the recycling of ascorbate from its oxidized form. Since TrxRs are able to reduce a number of substrates other than Trx, it is likely that additional biological effects will be discovered for TrxR. Furthermore, inhibiting TrxR with drugs may lead to new treatments for human diseases such as cancer, AIDS and autoimmune diseases. PMID:10657232

  8. Evaporation studies and phase stability of mercuric iodide

    NASA Astrophysics Data System (ADS)

    Piechotka, M.; Kaldis, E.

    1986-12-01

    Some aspects of the mass spectrometric analysis of a molecular beam of mercuric iodide relevant to the vapor growth of HgI 2 crystals are discussed. Mercuric iodide evaporates non-dissociatively in the temperature range of 35-150°C. Hydrocarbons and water are the main volatile impurities. It has been shown that hydrocarbons can be introduced in mercuric iodide not only by iodine but also by contamination by oil vapors from rotary vacuum pumps. Nonstoichiometry of mercuric iodine towards either excess of iodine or excess of mercury has been found. The nonstoichiometry is stabilized by the presence of organic impurities. The vapor pressure curve of HgI 2 shows an abrupt change in sublimation enthalpy at 67°C possibly due to a surface reconstruction at this temperature. As the growth temperature of the HgI 2 crystals is much higher, it is expected that the crystals will acquire an appreciable concentration of surface defects. This is in agreement with earlier observations of HgI 2 detector manufacturers.

  9. Evidence That the [beta] Subunit of Chlamydia trachomatis Ribonucleotide Reductase Is Active with the Manganese Ion of Its Manganese(IV)/Iron(III) Cofactor in Site 1

    SciTech Connect

    Dassama, Laura M.K.; Boal, Amie K.; Krebs, Carsten; Rosenzweig, Amy C.; Bollinger, Jr., J. Martin

    2014-10-02

    The reaction of a class I ribonucleotide reductase (RNR) begins when a cofactor in the {beta} subunit oxidizes a cysteine residue {approx}35 {angstrom} away in the {alpha} subunit, generating a thiyl radical. In the class Ic enzyme from Chlamydia trachomatis (Ct), the cysteine oxidant is the Mn{sup IV} ion of a Mn{sup IV}/Fe{sup III} cluster, which assembles in a reaction between O{sub 2} and the Mn{sup II}/Fe{sup II} complex of {beta}. The heterodinuclear nature of the cofactor raises the question of which site, 1 or 2, contains the Mn{sup IV} ion. Because site 1 is closer to the conserved location of the cysteine-oxidizing tyrosyl radical of class Ia and Ib RNRs, we suggested that the Mn{sup IV} ion most likely resides in this site (i.e., {sup 1}Mn{sup IV}/{sup 2}Fe{sup III}), but a subsequent computational study favored its occupation of site 2 ({sup 1}Fe{sup III}/{sup 2}Mn{sup IV}). In this work, we have sought to resolve the location of the Mn{sup IV} ion in Ct RNR-{beta} by correlating X-ray crystallographic anomalous scattering intensities with catalytic activity for samples of the protein reconstituted in vitro by two different procedures. In samples containing primarily Mn{sup IV}/Fe{sup III} clusters, Mn preferentially occupies site 1, but some anomalous scattering from site 2 is observed, implying that both {sup 1}Mn{sup II}/{sup 2}Fe{sup II} and {sup 1}Fe{sup II}/{sup 2}Mn{sup II} complexes are competent to react with O{sub 2} to produce the corresponding oxidized states. However, with diminished Mn{sup II} loading in the reconstitution, there is no evidence for Mn occupancy of site 2, and the greater activity of these 'low-Mn' samples on a per-Mn basis implies that the {sup 1}Mn{sup IV}/{sup 2}Fe{sup III}-{beta} is at least the more active of the two oxidized forms and may be the only active form.

  10. Large-area mercuric iodide x-ray imager

    NASA Astrophysics Data System (ADS)

    Zentai, George; Partain, Larry D.; Pavlyuchkova, Raisa; Virshup, Gary F.; Zuck, Asaf; Melekhov, Leonid; Dagan, O.; Vilensky, Alexander I.; Gilboa, Haim

    2002-05-01

    Single crystals of mercuric iodide have been studied for many years for nuclear detectors. We have investigated the use of x-ray photoconductive polycrystalline mercuric iodide coatings on amorphous silicon flat panel thin film transistor (TFT) arrays as x-ray detectors for radiographic and fluoroscopic applications in medical imaging. The mercuric iodide coatings were vacuum deposited by Physical Vapor Deposition (PVD). This coating technology is capable of being scaled up to sizes required in common medical imaging applications. Coatings were deposited on 4 inches X 4 inches TFT arrays for imaging performance evaluation and also on conductive-coated glass substrates for measurements of x-ray sensitivity, dark current and image lag. The TFT arrays used included pixel pitch dimensions of both 100 and 139 microns. Coating thickness between 150 microns and 250 microns were tested in the 25 kVp-100 kVp x-ray energy range utilizing exposures typical for both fluoroscopic, and radiographic imaging. X-ray sensitivities measured for the mercuric iodide samples and coated TFT detectors were superior to any published results for competitive materials (up to 7100 ke/mR/pixel for 100 micron pixels). It is believed that this higher sensitivity, can result in fluoroscopic imaging signal levels high enough to overshadow electronic noise. Image lag characteristics appear adequate for fluoroscopic rates. Resolution tests on resolution target phantoms showed that resolution is limited to the Nyquist frequency for the 139 micron pixel detectors. The ability to operate at low voltages gives adequate dark currents for most applications and allows low voltage electronics designs. Mercuric Iodide coated TFT arrays were found to be outstanding candidates for direct digital radiographic detectors for both static and dynamic (fluoroscopic) applications. Their high x-ray sensitivity, high resolution, low dark current, low voltage operation, and good lag characteristics provide a unique

  11. The absorption of mercuric ion into single human head hairs

    SciTech Connect

    Mackintosh, J.; Pate, B.D.

    1982-07-01

    The concentration patterns of radioactive Hg++, Cu++, and I- in individual hair shafts, after soaking in aqueous solutions of these tracers, were measured nondestructively to permit repeated experiments on a given shaft. The mercury concentrations generally increased from the root end to the distal end of a given shaft less steeply than those of copper, while iodide concentrations generally decreased. Concentration peaks and other pattern features for mercury were also relatively less intense, but there was some correlation of the position of such areas of increased mercury, copper, and iodine adsorption in a given shaft. At equilibrium after more than 100 h of soaking, the amount of mercury taken up at pH 8 by the hair was three to four times that at pH 3. The rate of absorption of mercury was higher at low pH values, and that of desorption higher at high pH values. The relative intensity of pattern features remained constant during absorption or desorption at a given pH, but changed if the pH was changed. These data are discussed in terms of the chemistry of the hair binding sites for cations and anions.

  12. Translocatable resistance to mercuric and phenylmercuric ions in soil bacteria

    SciTech Connect

    Radford, A.J.; Oliver, J.; Kelly, W.J.; Reaney, D.C.

    1981-08-01

    Of a sample of 42 grams-negative Hg-resistant bacteria, three (a Pseudomonas fluorescens, a Klebsiella sp. and a Citrobacter sp.) contained translocatable elements conferring resistance to Hgbj (all three) and to Hgbj and phenylmercuric acetate (P. fluorescens). The discovery of transposable phenylmercuric acetate resistance extends the range of known resistance ''transposons'' from heavy metals and antibiotics to organometallic compounds.

  13. Optical properties and surface morphology studies of palladium contacts on mercuric iodide single crystals

    NASA Astrophysics Data System (ADS)

    George, M. A.; Azoulay, M.; Burger, A.; Biao, Y.; Silberman, E.; Nason, D.

    1993-04-01

    Palladium is chemically suitable for electric contacts on mercuric iodide detectors for photon and nuclear radiation detection, so the understanding of palladium contacts is important for fundamental and practical scientific purposes. A study has been conducted on the surface morphology of evaporated contacts using atomic force microscopy (AFM) and optical transmission and reflection. Evaporated palladium coatings are typically nonuniform and may deposit selectively on mercuric iodide surface defects. Reflection measurements show that coating thickness and surface treatment affect intensity, position, and shape of a reflected peak characteristic of the mercuric iodide structure. Results indicate that the band gap energy in the surface of the mercuric iodide is lowered by palladium contacts.

  14. Direct vapor/solid synthesis of mercuric iodide using compounds of mercury and iodine

    DOEpatents

    Skinner, Nathan L.

    1990-01-01

    A process is disclosed for producing high purity mercuric iodide by passing a gaseous source of a mercuric compound through a particulate bed of a low vapor pressure iodide compound which is maintained at an elevated temperature which is the lower of either: (a) just below the melting or volatilization temperature of the iodide compound (which ever is lower); or (b) just below the volatilization point of the other reaction product formed during the reaction; to cause the mercuric compound to react with the iodide compound to form mercuric iodide which then passes as a vapor out of the bed into a cooler condensation region.

  15. Sodium selenite and vitamin E in preventing mercuric chloride induced renal toxicity in rats.

    PubMed

    Aslanturk, Ayse; Uzunhisarcikli, Meltem; Kalender, Suna; Demir, Filiz

    2014-08-01

    This study aims to investigate improving effects of sodium selenite and/or vitamin E on mercuric chloride-induced kidney impairments in rats. Wistar male rats were exposed either to sodium selenite (0.25mg/kgday), vitamin E (100mg/kgday), sodium selenite+vitamin E, mercuric chloride (1mg/kgday), sodium selenite+mercuric chloride, vitamin E+mercuric chloride and sodium selenite+vitamin E+mercuric chloride for 4weeks. Mercuric chloride exposure resulted in an increase in the uric acid, creatinine, blood urea nitrogen and malondialdehyde (MDA) levels and a decrease in the superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) activities. Histopathological changes were detected in kidney tissues in mercuric chloride-treated groups. A significant decrease in the uric acid, creatinine, blood urea nitrogen and MDA levels and a significant increase in the SOD, CAT and GPx activities were observed in the supplementation of sodium selenite and/or vitamin E to mercuric chloride-treated groups. Conclusively, sodium selenite, vitamin E and vitamin E+sodium selenite significantly reduce mercuric chloride induced nephrotoxicity in rats, but not protect completely.

  16. Growth of mercuric iodide single crystals from dimethylsulfoxide

    DOEpatents

    Carlston, Richard C.

    1976-07-13

    Dimethylsulfoxide is used as a solvent for the growth of red mercuric iodide (HgI.sub.2) crystals for use in radiation detectors. The hygroscopic property of the solvent allows controlled amounts of water to enter into the solvent phase and diminish the large solubility of HgI.sub.2 so that the precipitating solid collects as well-defined euhedral crystals which grow into a volume of several cc.

  17. Elemental impurity analysis of mercuric iodide by ICP/MS

    SciTech Connect

    Cross, E.S.; Mroz, E.; Olivares, J.A.

    1993-06-01

    A method has been developed to analyze mercuric iodide (HgI{sub 2}) for elemental contamination using Inductively Coupled Plasma/Mass Spectroscopy (ICP/MS). This paper will discuss the ICP/MS method, the effectiveness of purification schemes for removing impurities from HgI{sub 2}, as well as preliminary correlations between HgI{sub 2} detector performance and elemental contamination levels.

  18. Elemental impurity analysis of mercuric iodide by ICP/MS

    SciTech Connect

    Cross, E.S. . Santa Barbara Operations); Mroz, E.; Olivares, J.A. )

    1993-01-01

    A method has been developed to analyze mercuric iodide (HgI[sub 2]) for elemental contamination using Inductively Coupled Plasma/Mass Spectroscopy (ICP/MS). This paper will discuss the ICP/MS method, the effectiveness of purification schemes for removing impurities from HgI[sub 2], as well as preliminary correlations between HgI[sub 2] detector performance and elemental contamination levels.

  19. Phonon dispersion in red mercuric iodide

    SciTech Connect

    Sim, H.; Chang, Y. ); James, R.B. )

    1994-02-15

    We present theoretical studies of phonon modes of undoped HgI[sub 2] in its red tetragonal form. A rigid-ion model including the Coulomb interaction is used which gives the best fit to the neutron scattering, infrared reflectivity, and Raman scattering data. The calculated sound velocities are also in accord with experiment.

  20. Horizontal Ampoule Growth and Characterization of Mercuric Iodide at Controlled Gas Pressures for X-Ray and Gamma Ray Spectrometers

    SciTech Connect

    McGregor, Douglas S.; Ariesanti, Elsa; Corcoran, Bridget

    2004-04-30

    The project developed a new method for producing high quality mercuric iodide crystals of x-ray and gamma spectrometers. Included are characterization of mercuric iodide crystal properties as a function of growth environment and fabrication and demonstration of room-temperature-operated high-resolution mercuric iodide spectrometers.

  1. Mercuric iodate precipitation from radioiodine-containing off-gas scrubber solution

    DOEpatents

    Partridge, Jerry A.; Bosuego, Gail P.

    1982-01-01

    Mercuric nitrate-nitric acid scrub solutions containing radioiodine may be reduced in volume without excessive loss of volatile iodine. The use of concentrated nitric acid during an evaporation process oxidizes the mercury-iodide complex to a less volatile mercuric iodate precipitate.

  2. Application of Mercuric Iodide Detectors to the Monitoring and Evaluation of Stored Special Nuclear Materials

    DTIC Science & Technology

    2001-01-01

    Application of Mercuric Iodide Detectors to the Monitoring and Evaluation of Stored Special Nuclear Materials L. van den Berg, A.E. Proctor and K.R...2001 to 00-00-2001 4. TITLE AND SUBTITLE Application of Mercuric Iodide Detectors to the Monitoring and Evaluation of Stored Special Nuclear

  3. Ameliorative stroke of selenium against toxicological effects of mercuric chloride in liver of freshwater catfish Heteropneustes fossilis (Bloch).

    PubMed

    Kothari, Suresh; Choughule, Neha

    2015-07-08

    Mercury, a prevalent and unrelenting toxin, occurs in a variety of forms in freshwater as well as, in marine life. Mercury is an important inducer of oxidative stress in fish leading to formation of reactive oxygen species. Selenium is an essential micronutrient for animals and has antagonistic effect against mercuric toxicity in fishes. Present study has been made to evaluate toxic effect of HgCl2 (0.15 mg/L) on liver of freshwater catfish Heteropneustes fossilis (Bl.). Protective ability of selenium has been investigated by simultaneous exposure of fish with sodium selenite (0.15 mg/L) along with mercuric chloride. For present study Fishes were divided into three groups of ten fishes each the first group served as control, while the second group fish were exposed to HgCl2 . Animals of third group were treated with HgCl2 and Na2 SeO3 . Results reveal that mercury induced lipid peroxidation and in response to this, antioxidants reduced glutathione (GSH) and Catalase (CAT) were reduced whereas, Glutathione reductase (GR) level was enhanced. These antioxidants scavenge the reactive oxygen radicals. Hg induced histopathological damage and elevation in alkaline phosphatase (ALP) and transaminases and reduction in protein and glucose contents were evidently seen in catfish liver. Intriguingly, results indicate that under stress of mercury, the fish actively generate oxidative stress and antioxidant responses, which can be used as biomarkers of pollution. Simultaneous exposure to Selenium along with Hg suppressed Hg uptake and lipid peroxidation. Histological architecture and all biochemical parameters were maintained near normal in the presence of selenium in liver of the catfish.

  4. Development of mercuric iodide uncooled x ray detectors and spectrometers

    NASA Technical Reports Server (NTRS)

    Iwanczyk, Jan S.

    1990-01-01

    The results obtained in the development of miniature, lowpower, light weight mercuric iodide, HgI2, x ray spectrometers for future space missions are summarized. It was demonstrated that HgI2 detectors can be employed in a high resolution x ray spectrometer, operating in a scanning electron microscope. Also, the development of HgI2 x ray detectors to augment alpha backscattering spectrometers is discussed. These combination instruments allow for the identification of all chemical elements, with the possible exception of hydrogen, and their respective concentrations. Additionally, further investigations of questions regarding radiation damage effects in the HgI2 x ray detectors are reported.

  5. Low-temperature photoluminescence studies of mercuric-iodide photodetectors

    SciTech Connect

    James, R.B. ); Bao, X.J. ); Schlesinger, T.E.; Markakis, J.M.; Cheng, A.Y.; Ortale, C.

    1989-09-15

    Mercuric-iodide (HgI{sub 2} ) photodetectors with sputtered indium-tin-oxide (ITO) entrance electrodes were studied using low-temperature photoluminescence spectroscopy. The photoluminescence spectrum obtained on each photodetector was found to differ for points beneath the ITO contact and points adjacent to it, indicating that the contact fabrication process introduces new carrier traps and radiative recombination centers within the ITO-HgI{sub 2} interfacial region. In particular, a new broad band was observed in the spectra taken from points beneath the ITO electrode. Photocurrent-versus-position measurements showed that the intensity of this broad band was enhanced in regions having relatively poor photoresponse.

  6. Elemental impurity analysis of mercuric iodide by ICP/MS

    SciTech Connect

    Cross, E.S.; Mroz, E.; Olivares, J.A.

    1994-06-01

    A method has been developed to analyze mercuric iodide (HgI{sub 2}) for elemental contamination using Inductively Coupled Plasma/Mass Spectroscopy (ICP/MS). This paper discusses the ICP/MS method, the effectiveness of purification schemes for removing impurities from HgI{sub 2}, as well as preliminary correlations between HgI{sub 2} detector performance and elemental contamination levels. The purified HgI{sub 2} is grown into a single crystal by physical vapor transport. The crystal are cut into slices and they are fabricated into room temperature radiation detectors and photocells. Crystals that produce good resolution gamma detector do not necessarily make good resolution photocells or x-ray detectors. Many factors other than elemental impurities may contribute to these differences in performance.

  7. Mercuric Iodide Photocell Technology for Room Temperature Readout of Scintillators

    SciTech Connect

    Warnick Kernan et al.

    2007-08-31

    Mercuric iodide (HgI2) is a well known material for the direct detection of gamma rays; however, the largest volume achievable is limited by thickness of the detector, which needs to be a small fraction of the average trapping length for electrons. We are reporting here preliminary results in using HgI2 crystals to fabricate photocells used in the readout of various scintillators. The optical spectral response and efficiency of these photocells were measured and will be reported. Preliminary nuclear response from a HgI2 photocell that was optically matched to a Ce3+ :LaBr3 scintillator will also be presented and discussed. Further improvements will be sought by optimizing the transparent contact technology.

  8. Comparison of transparent conducting electrodes on mercuric iodide photocells

    NASA Astrophysics Data System (ADS)

    Cheng, A. Y.; Markakis, J. M.

    Three materials have been developed and tested which are suitable as transparent conducting electrodes on mercuric iodide; aqueous ionic contacts of NaCl and LiCl, polyvinyl alcohol/phosphoric acid, and indium--tin--oxide (ITO). Polyvinyl alcohol/phosphoric acid is a conducting polymer and ITO is a wide band gap semiconductor. Photocell dimensions were in the range of 0.5 to 3.8 cm diam by about 1 mm thick. Photocells with these electrodes were evaluated for their spectral response in the range of 300 to 650 nm, response uniformity over the electrode activities area, leakage current and reliability. All units showed better than 75 percent quantum efficiency in the range 350 to 550 nm. Photodetector leakage currents ranged from 25 to 200 pA and have shown long term stability up to 1 year.

  9. Comparison of transparent conducting electrodes on mercuric iodide photocells

    NASA Astrophysics Data System (ADS)

    Markakis, J. M.; Cheng, A. Y.

    1989-11-01

    Three materials have been developed and tested which are suitable as transparent conducting electrodes on mercuric iodide: aqueous ionic contacts of NaCl and LiCl, polyvinyl alcohol/phosphoric acid, and indium-tin-oxide (ITO). Polyvinyl alcohol/phosphoric acid is a conducting polymer and ITO is a wide band gap semiconductor. Photocell dimensions were in the range of 0.5 to 3.8 cm diameter by about 1 mm thick. Photocells with these electrodes were evaluated for their spectral response in the range of 300 to 650 nm, response uniformity over the electrode active area and reliability. All units showed better than 75% quantum efficiency in the range of 350 to 550 nm. Photodetector leakage currents ranged from 25 to 200 pA and have shown long-term stability up to one year.

  10. Electronic characterization of mercuric iodide gamma ray spectrometers

    SciTech Connect

    Gerrish, V.M.

    1993-06-01

    During the past four years the yield of high resolution mercuric iodide (HgI{sub 2}) gamma ray spectrometers produced at EG&G/EM has increased dramatically. Data is presented which demonstrates a strong correlation between starting material and spectrometer performance. Improved spectrometer yields are attributed to the method of HgI{sub 2} synthesis and to material purification procedures. Data is presented which shows that spectrometer performance is correlated with hole mobility-lifetime products. In addition, the measurement of Schottky barrier heights on HgI{sub 2} spectrometers has been performed using I-V curves and the photoelectric method. Barrier heights near 1.1 eV have been obtained using various contacts and contact deposition methods. These data suggest the pinning of the Fermi level at midgap at the HgI{sub 2} surface, probably due to surface states formed prior to contact deposition.

  11. Electronic characterization of mercuric iodide gamma ray spectrometers

    SciTech Connect

    Gerrish, V.M.

    1993-01-01

    During the past four years the yield of high resolution mercuric iodide (HgI[sub 2]) gamma ray spectrometers produced at EG G/EM has increased dramatically. Data is presented which demonstrates a strong correlation between starting material and spectrometer performance. Improved spectrometer yields are attributed to the method of HgI[sub 2] synthesis and to material purification procedures. Data is presented which shows that spectrometer performance is correlated with hole mobility-lifetime products. In addition, the measurement of Schottky barrier heights on HgI[sub 2] spectrometers has been performed using I-V curves and the photoelectric method. Barrier heights near 1.1 eV have been obtained using various contacts and contact deposition methods. These data suggest the pinning of the Fermi level at midgap at the HgI[sub 2] surface, probably due to surface states formed prior to contact deposition.

  12. Electrical and photomechanical effects of plastic deformation of mercuric iodide

    NASA Astrophysics Data System (ADS)

    Marschall, J.; Milstein, F.; Georgeson, G.; Gerrish, V.

    The effects of bulk plastic deformation of mercuric iodide (HgI2), upon some of the electronic properties relevant to the performance of HgI2 as a radiation detector were examined experimentally. Hole lifetimes, as well as hole and electron mobilities, were measured at various stages of sample deformation. Hole lifetimes were found to decrease by a factor of 2 under strains of several percent; carrier mobilities varied within experimental error, except during creep loading where electron and hole mobilities decreased by about 65 percent and 25 percent, respectively. Additionally, dark current measurements were made on specimens with varying degrees of accumulated plastic damage caused by c plane shear. Dark current values did not strongly reflect the extent of bulk plastic damage in deformed specimens.

  13. Incorporation of defects during processing of mercuric iodide detectors

    SciTech Connect

    Bao, X.J.; Schlesinger, T.E. ); James, R.B.; Stulen, R.H. ); Ortale, C.; Cheng, A.Y. )

    1990-07-01

    The effects of chemical etching in KI solution, heating, and vacuum exposures of HgI{sub 2} were individually studied by low-temperature photoluminescence (PL) spectroscopy. Each of these processing steps is important in the manufacturing of mercuric iodide detectors and may be responsible for the incorporation of carrier traps both in the near-surface region and in the bulk. The results of etching experiments showed that the near-surface region has a different defect structure than the bulk, which appears to result from iodine deficiency. Bulk heating at 100 {degree}C also modifies the defect structure of the crystal. Vacuum exposure has an effect similar to chemical etching, but it does not cause significant degradation of the stoichiometry for recently KI-etched specimens. These studies suggest that some features in the PL spectra of HgI{sub 2} are associated with stoichiometry of the specimens.

  14. Incorporation of defects during processing of mercuric iodide detectors

    NASA Astrophysics Data System (ADS)

    Bao, X. J.; Schlesinger, T. E.; James, R. B.; Stulen, R. H.; Ortale, C.; Cheng, A. Y.

    1990-07-01

    The effects of chemical etching in KI solution, heating, and vacuum exposures of HgI2 were individually studied by low-temperature photoluminescence (PL) spectroscopy. Each of these processing steps is important in the manufacturing of mercuric iodide detectors and may be responsible for the incorporation of carrier traps both in the near-surface region and in the bulk. The results of etching experiments showed that the near-surface region has a different defect structure than the bulk, which appears to result from iodine deficiency. Bulk heating at 100 °C also modifies the defect structure of the crystal. Vacuum exposure has an effect similar to chemical etching, but it does not cause significant degradation of the stoichiometry for recently KI-etched specimens. These studies suggest that some features in the PL spectra of HgI2 are associated with stoichiometry of the specimens.

  15. Investigation of copper electrodes for mercuric iodide detector applications

    SciTech Connect

    Bao, X.J.; Schlesinger, T.E. ); James, R.B.; Stulen, R.H. ); Ortale, C.; van den Berg, L. )

    1990-06-15

    Copper diffusion in mercuric iodide was studied by low-temperature photoluminescence (PL) spectroscopy and Auger electron spectroscopy. A broad radiative emission band at a wavelength of about 6720 A in the PL spectra was found to be related to Cu incorporation in the crystal. PL spectra obtained from surface doping experiments indicate that Cu is a rapid diffuser in HgI{sub 2} bulk material. Auger electron spectroscopy performed as a function of depth from the crystal surface confirms the rapid bulk diffusion process of Cu in HgI{sub 2}. Fabrication of HgI{sub 2} nuclear detectors with Cu electrodes indicates that Cu is not acceptable as an electrode material, which is consistent with the fact that it diffuses easily into the bulk crystal and introduces new radiative recombination centers.

  16. Electrical and photomechanical effects of plastic deformation of mercuric iodide

    SciTech Connect

    Marschall, J.; Milstein, F. . Dept. of Materials California Univ., Santa Barbara, CA . Dept. of Mechanical Engineering); Georgeson, G. ); Gerrish, V. . Santa Barbara Operations)

    1991-01-01

    The effects of bulk plastic deformation of mercuric iodide (HgI{sub 2}), upon some of the electronic properties relevant to the performance of HgI{sub 2} as a radiation detector were examined experimentally. Hole lifetimes, as well as hole and electron mobilities, were measured at various stages of sample deformation. Hole lifetimes were found to decrease by a factor of 2 under strains of several percent; carrier mobilities varied within experimental error, except during creep loading where electron and hole mobilities decreased by about 65 % and 25 %, respectively. Additionally, dark current measurements were made on specimens with varying degrees of accumulated plastic damage caused by c plane shear. Dark current values did not strongly reflect the extent of bulk plastic damage in deformed specimens. 16 refs., 4 figs., 1 tab.

  17. Mercuric iodide crystals obtained by solvent evaporation using ethanol

    NASA Astrophysics Data System (ADS)

    Ugucioni, J. C.; Ghilardi Netto, T.; Mulato, M.

    2010-04-01

    Millimeter-sized mercuric iodide crystals were fabricated by the solvent evaporation technique using pure ethanol as a solvent. Three different conditions for solution evaporation were tested: (i) in the dark at room temperature; (ii) in the presence of light at room temperature and (iii) in an oven at 40 °C. Morphology, structure, optical and electrical properties were investigated using several techniques. Crystals fabricated in the dark show better properties and stability than others, possibly because the larger the energy of the system, the larger the number of induced growth defects. The crystals fabricated in the dark have adequate structure for higher resistivity and activation energy close to half the optical band-gap, as desired. With proper encapsulation these crystals might be good candidates for the development of ionizing radiation sensors.

  18. Destructive and regenerative changes in the albino rat kidney during mercuric chloride necrotizing nephrosis

    SciTech Connect

    Andreev, V.P.

    1985-08-01

    This paper describes the results of a morphological analysis of destructive and regenerative changes observed during a study of serial semithin sections of the kidneys of albino rats with mercuric chloride necrotizing nephrosis. The results of this investigation indicate that injury to the epithelium of the urinary tubules by mercuric chloride is heterogenous in depth, and this has a substantial influence on the viability of the animals and on the subsequent process of repair of the damage.

  19. Photochemical reactions of divalent mercury with thioglycolic acid: formation of mercuric sulfide particles.

    PubMed

    Si, Lin; Ariya, Parisa A

    2015-01-01

    Mercury (Hg) is a key toxic global pollutant. Studies in aquatic environment have suggested that thiols could be important for mercury speciation. Thioglycolic acid has been detected in various natural water systems and used as a model compound to study the complicated interaction between mercury and polyfunctional dissolved organic matter (DOM). We herein presented the first evidence for mercury particle formation during kinetic and product studies on the photochemistry of divalent mercury (Hg(2+)) with thioglycolic acid at near environmental conditions. Mercuric sulfide (HgS) particles formed upon photolysis were identified by high-resolution transmission electron microscopy coupled with energy dispersive spectrometry and select area electron diffraction. Kinetic data were obtained using UV-visible spectrophotometry and cold vapour atomic fluorescent spectrometry. The apparent first-order reaction rate constant under our experimental conditions was calculated to be (2.3±0.4)×10(-5) s(-1) at T=296±2 K and pH 4.0. It was found that (89±3)% of the reactants undergo photoreduction to generate elemental mercury (Hg(0)). The effects of ionic strengths, pH and potassium ion were also investigated. The formation of HgS particles pointed to the possible involvement of heterogeneous processes. Our kinetic results indicated the importance of weak binding sites on DOM to Hg in photoreduction of Hg(2+) to Hg(0). The potential implications of our data on environmental mercury transformation were discussed.

  20. Soluble ascorbate free radical reductase in the human lens.

    PubMed

    Bando, M; Obazawa, H

    1994-01-01

    A major and a minor ascorbate free radical (AFR) reductase were separated from the soluble fraction in the human lens cortex by DEAE-cellulose ion-exchange column chromatography. These AFR reductases also exhibited diaphorase activity using dichlorophenolindophenol and ferricyanide as electron acceptors. The major AFR reductase was partially purified by 5'AMP-Sepharose 4B affinity column chromatography. This partially purified AFR reductase showed a single band of diaphorase activity in native polyacrylamide disc gel electrophoresis. This activity band corresponded to the major protein observed in protein staining by Coomassie Brilliant Blue. However, the protein staining by Coomassie Brilliant Blue showed this activity band surrounded by diffused staining. Molecular weight of the partially purified AFR reductase was determined to be 32 kDa by gel filtration, and the apparent Km value for AFR was about 15 microM. This major lens AFR reductase could be distinguished from soluble Neurospora, Euglena and cucumber AFR reductases, and from two ubiquitous enzymes with reduction activity of AFR and/or foreign compounds, ie, NADH-cytochrome b5 reductase and DT-diaphorase, by their molecular weights, Km values and/or ion-exchange chromatographic behaviors.

  1. Mercuric 5-Nitrotetrazole, a Possible Replacement for Lead Azide in Australian Ordnance. Part 1. An Assessment of Preparation Methods

    DTIC Science & Technology

    1983-08-01

    nitric acid (0.4 ml) in 1.0 M aqueous mercuric nitrate (25.5 ml) [from dissolution of red mercuric oxide (54.0 q) in 35% nitric acid (200 ml...R MRL-R-901 MERCURIC 5-NITROTETRAZOLE, A POSSIBLE REPLACEMENT FOR LEAD AZIDE IN AUSTRALIAN ORDNANCE PART 1 AN ASSESSMENT OF PREPARATION METHQD.. L.D...Distribut iMoR/Ava,’ ,. MRL-R-901 MERCURIC 5-NITROTETRAZOLE, A POSSIBLE REPLACEMENT FOR LEAD AZIDE IN AUSTRALIAN ORDNANCE \\ PART 1 AN ASSESSMENT OF

  2. The tyrosyl free radical in ribonucleotide reductase.

    PubMed Central

    Gräslund, A; Sahlin, M; Sjöberg, B M

    1985-01-01

    The enzyme, ribonucleotide reductase, catalyses the formation of deoxyribonucleotides from ribonucleotides, a reaction essential for DNA synthesis in all living cells. The Escherichia coli ribonucleotide reductase, which is the prototype of all known eukaryotic and virus-coded enzymes, consists of two nonidentical subunits, proteins B1 and B2. The B2 subunit contains an antiferromagnetically coupled pair of ferric ions and a stable tyrosyl free radical. EPR studies show that the tyrosyl radical, formed by loss of ferric ions and a stable tyrosyl free radical. EPR studies show that the tyrosyl radical, formed by loss of an electron, has its unpaired spin density delocalized in the aromatic ring of tyrosine. Effects of iron-radical interaction indicate a relatively close proximity between the iron center and the radical. The EPR signal of the radical can be studied directly in frozen packed cells of E. coli or mammalian origin, if the cells are made to overproduce ribonucleotide reductase. The hypothetic role of the tyrosyl free radical in the enzymatic reaction is not yet elucidated, except in the reaction with the inhibiting substrate analogue 2'-azido-CDP. In this case, the normal tyrosyl radical is destroyed with concomitant appearance of a 2'-azido-CDP-localized radical intermediate. Attempts at spin trapping of radical reaction intermediates have turned out negative. In E. coli the activity of ribonucleotide reductase may be regulated by enzymatic activities that interconvert a nonradical containing form and the fully active protein B2. In synchronized mammalian cells, however, the cell cycle variation of ribonucleotide reductase, studied by EPR, was shown to be due to de novo protein synthesis. Inhibitors of ribonucleotide reductase are of medical interest because of their ability to control DNA synthesis. One example is hydroxyurea, used in cancer therapy, which selectively destroys the tyrosyl free radical. PMID:3007085

  3. Quinone Reductase 2 Is a Catechol Quinone Reductase

    SciTech Connect

    Fu, Yue; Buryanovskyy, Leonid; Zhang, Zhongtao

    2008-09-05

    The functions of quinone reductase 2 have eluded researchers for decades even though a genetic polymorphism is associated with various neurological disorders. Employing enzymatic studies using adrenochrome as a substrate, we show that quinone reductase 2 is specific for the reduction of adrenochrome, whereas quinone reductase 1 shows no activity. We also solved the crystal structure of quinone reductase 2 in complexes with dopamine and adrenochrome, two compounds that are structurally related to catecholamine quinones. Detailed structural analyses delineate the mechanism of quinone reductase 2 specificity toward catechol quinones in comparison with quinone reductase 1; a side-chain rotational difference between quinone reductase 1 and quinone reductase 2 of a single residue, phenylalanine 106, determines the specificity of enzymatic activities. These results infer functional differences between two homologous enzymes and indicate that quinone reductase 2 could play important roles in the regulation of catecholamine oxidation processes that may be involved in the etiology of Parkinson disease.

  4. Complexation of mercury(I) and mercury(II) by 18-crown-6: hydrothermal synthesis of the mercuric nitrite complex.

    PubMed

    Williams, Neil J; Hancock, Robert D; Riebenspies, Joseph H; Fernandes, Manuel; de Sousa, Alvaro S

    2009-12-21

    A dimercury(I) 18-crown-6 complex is isolated, and its possible role in the hydrothermal preparation of the mercuric nitrite complex is discussed. The reported structures are of [Hg(2)(18-crown-6)(2)(H(2)O)(2)](ClO(4))(2) (1), monoclinic, C2/c, a = 21.0345(9), b = 12.1565(5), c = 16.8010(7) A, beta = 113.2000(10) degrees , V = 3948.7(3) A(3), Z = 16, R = 0.0230; [Hg(18-crown-6)](NO(2))(2) (2), monoclinic, P2(1)/c, a = 8.027(5), b = 14.437(9), c = 7.827(5) A, beta = 95.165(11) degrees , V = 905.6(10) A(3), Z = 2, R = 0.0175. The complex cation in compound 1 consists of a mercurous dimer exhibiting a Hg-Hg bond length of 2.524(2) A. Non-bonding interactions between adjacent crown ether macrocycles across the Hg-Hg bond result in large variations in mercury to oxygen distances within equatorial coordination sites. At low pH compound 1 is proposed to be preferentially formed under hydrothermal conditions affording compound 2 upon disproportionation. Nitrite ions ligate via a unidentate nitrito (cis to metal) coordination mode as interpreted using vibrational (infrared) spectroscopy. The conformation adopted by 18-crown-6 in compounds 1 and 2 is closely related to a D(3d) conformation as evidenced by X-ray crystallography. Band splitting readily observed in vibrational spectra of the metal free crown ether, attributed to vibrational modes of oxyethylene fragments, is absent in spectra of 1 and 2 confirming a regular D(3d) macrocyclic orientation. Short Hg-O bonds observed for axially coordinated water molecules in 1 and coordinated nitrite ligands in 2, illustrate the prevalence of relativistic effects commonly observed in mercury complexes.

  5. Pulsed laser photofragment emission for detection of mercuric chloride

    NASA Astrophysics Data System (ADS)

    Hoops, Alexandra A.; Reichardt, Thomas A.

    2006-08-01

    The viability of pulsed laser photofragment emission (PFE) is evaluated for the in situ measurement of vapor-phase mercuric chloride (HgCl2) concentration in combustion flue gas. Dispersed emissions from both the Hg (63P1) and HgCl (B2Σ+) photoproducts are presented, and the dependence of the HgCl2 PFE signal originating from Hg (63P1) on the collisional environment is examined for buffer-gas mixtures of N2, O2, and CO2. Integrated PFE intensity measurements as a function of buffer gas pressure support the assumption that the primary effect of the relevant flue gas constituents is to quench emission from Hg (63P1). The quenching rate constants for PFE from HgCl2 were measured to be 1.37 (±0.16)×105 Torr-1 s-1 for N2, 9.35 (±0.25)×106 Torr-1 s-1 for O2, and 1.49 (±0.29)×106 Torr-1 s-1 for CO2. These values are in good accord with literature values for the quenching of Hg (63P1). The emission cross section for Hg (63P1) generated by photodissociation of HgCl2 in 760 Torr N2 is found to be 1.0 (±0.2)×10-25 m2 by comparing the PFE signal to N2 Raman scattering.

  6. Polycrystalline Mercuric Iodide Films on CMOS Readout Arrays

    PubMed Central

    Hartsough, Neal E.; Iwanczyk, Jan S.; Nygard, Einar; Malakhov, Nail; Barber, William C.; Gandhi, Thulasidharan

    2009-01-01

    We have created high-resolution x-ray imaging devices using polycrystalline mercuric iodide (HgI2) films grown directly onto CMOS readout chips using a thermal vapor transport process. Images from prototype 400×400 pixel HgI2-coated CMOS readout chips are presented, where the pixel grid is 30 μm × 30 μm. The devices exhibited sensitivity of 6.2 μC/Rcm2 with corresponding dark current of ∼2.7 nA/cm2, and a 80 μm FWHM planar image response to a 50 μm slit aperture. X-ray CT images demonstrate a point spread function sufficient to obtain a 50 μm spatial resolution in reconstructed CT images at a substantially reduced dose compared to phosphor-coated readouts. The use of CMOS technology allows for small pixels (30 μm), fast readout speeds (8 fps for a 3200×3200 pixel array), and future design flexibility due to the use of well-developed fabrication processes. PMID:20161098

  7. Optical anisotropic-dielectric response of mercuric iodide

    SciTech Connect

    Yao, H.; Johs, B.; James, R.B.

    1997-10-01

    Anisotropic optical properties of mercuric iodide (HgI{sub 2}) were studied by variable-angle spectroscopic ellipsometry (VASE). Angular-dependent polarized reflectance and transmittance at three special optical-axis configurations, concerning the uniaxial anisotropic nature of the crystal, were derived to facilitate the VASE analysis. Two surface orientations of this tetragonal crystal were selected, i.e., an a-plane and a c-plane sample. Room-temperature multiple-angle spectroscopic ellipsometry measurements from both samples with three different optical configurations along with polarized transmission measurements were jointly analyzed by the VASE analysis through multiple-sample, multiple-model methods. Anisotropic dielectric functions of single-crystal HgI{sub 2}, {var_epsilon}{sub {perpendicular}}({omega}) and {var_epsilon}{sub {parallel}}({omega}), for optical electric-field vector oriented perpendicular and parallel to the c axis, respectively, were obtained in the range 1.24{endash}5.1 eV. Different absorption energy-band edges, at room temperature, were observed from the ordinary and extraordinary dielectric responses at 2.25 and 2.43 eV, respectively. This is consistent with the results related to the optical transitions between the conduction band and the heavy- and light-hole valence band indicated by theoretical studies. A surface model related to the surface roughness and defects of HgI{sub 2} was established and characterized by the VASE analysis. {copyright} {ital 1997} {ital The American Physical Society}

  8. Fabrication and performance of mercuric iodide pixellated detectors

    NASA Astrophysics Data System (ADS)

    van den Berg, Lodewijk; Bastian, Lloyd F.; Zhang, Feng; Lenos, Howard; Capote, M. Albert

    2007-09-01

    The radiation detection efficiency and spectral resolution of mercuric iodide detectors can be improved significantly by increasing the volume of the detectors and by using a pixellated anode structure. Detector bodies with a thickness of nominally 10 mm and an active area of approximately 14 mm x 14 mm have been used for these experiments. The detectors were cut from single crystals grown by the physical vapor transport method. The cut surfaces were polished and etched using a string saw and potassium iodide solutions. The Palladium contacts were deposited by magnetron sputtering through stainless steel masks. The cathode contact is continuous; the anode contacts consist of an array of 11 x 11 pixels surrounded by a guard ring. The resistance between a pixel and its surrounding contacts should be larger than 0.25 Gohm. The detector is mounted on a substrate that makes it possible to connect the anode pixels to an ASIC, and is conditioned so that it is stable for all pixels at a bias of -3000 Volts. Under these conditions the spectral resolution for Cs-137 gamma rays (662 keV) is approximately 5% FWHM. When depth sensing correction methods are applied, the resolution improves to about 2% FWHM or better. It is expected that the performance of the devices can be improved by the careful selection of crystal parts that are free of structural defects. Details of the fabrication technologies will be described. The effects of material inhomogeneities and transport properties of the charge carriers will be discussed.

  9. Reduction of Flavodoxin by Electron Bifurcation and Sodium Ion-dependent Reoxidation by NAD+ Catalyzed by Ferredoxin-NAD+ Reductase (Rnf).

    PubMed

    Chowdhury, Nilanjan Pal; Klomann, Katharina; Seubert, Andreas; Buckel, Wolfgang

    2016-06-03

    Electron-transferring flavoprotein (Etf) and butyryl-CoA dehydrogenase (Bcd) from Acidaminococcus fermentans catalyze the endergonic reduction of ferredoxin by NADH, which is also driven by the concomitant reduction of crotonyl-CoA by NADH, a process called electron bifurcation. Here we show that recombinant flavodoxin from A. fermentans produced in Escherichia coli can replace ferredoxin with almost equal efficiency. After complete reduction of the yellow quinone to the blue semiquinone, a second 1.4 times faster electron transfer affords the colorless hydroquinone. Mediated by a hydrogenase, protons reoxidize the fully reduced flavodoxin or ferredoxin to the semi-reduced species. In this hydrogen-generating system, both electron carriers act catalytically with apparent Km = 0.26 μm ferredoxin or 0.42 μm flavodoxin. Membrane preparations of A. fermentans contain a highly active ferredoxin/flavodoxin-NAD(+) reductase (Rnf) that catalyzes the irreversible reduction of flavodoxin by NADH to the blue semiquinone. Using flavodoxin hydroquinone or reduced ferredoxin obtained by electron bifurcation, Rnf can be measured in the forward direction, whereby one NADH is recycled, resulting in the simple equation: crotonyl-CoA + NADH + H(+) = butyryl-CoA + NAD(+) with Km = 1.4 μm ferredoxin or 2.0 μm flavodoxin. This reaction requires Na(+) (Km = 0.12 mm) or Li(+) (Km = 0.25 mm) for activity, indicating that Rnf acts as a Na(+) pump. The redox potential of the quinone/semiquinone couple of flavodoxin (Fld) is much higher than that of the semiquinone/hydroquinone couple. With free riboflavin, the opposite is the case. Based on this behavior, we refine our previous mechanism of electron bifurcation.

  10. Plasmid profiles of mercuric chloride tolerant rhizobia from horse gram (Macrotyloma uniflorum).

    PubMed

    Edulamudi, Prabhavati; Johnson, Antony A M; Divi, Venkata Ramana Sai Gopal; Konada, Veera Mallaiah

    2012-03-01

    Thirty two rhizobia were isolated from the fresh healthy root nodules of horse gram. They were found to be highly salt tolerant. They were identified as rhizobia by cultural, biochemical and 16S rRNA sequence. The sequences of the four selected isolates were deposited in the NCBI GenBank. The obtained accession numbers were GQ483457, GQ483458, GQ483459 and GQ483460. All the rhizobia were able to grow at 10 ppm mercuric chloride concentration. Four isolates HGR-11, 16, 30 and 31 were used to study the effect of different concentrations of mercuric chloride on the growth of rhizobia. These isolates were able to grow at 30 ppm concentration also. In these isolates, HGR-11 and HGR-30 showed maximum growth at 20 ppm than at control. These isolates contained one mega plasmid (-22 kb) at 20 ppm mercuric chloride concentration.

  11. A bacterial view of the periodic table: genes and proteins for toxic inorganic ions.

    PubMed

    Silver, Simon; Phung, Le T

    2005-12-01

    Essentially all bacteria have genes for toxic metal ion resistances and these include those for Ag+, AsO2-, AsO4(3-), Cd2+ Co2+, CrO4(2-), Cu2+, Hg2+, Ni2+, Pb2+, TeO3(2-), Tl+ and Zn2+. The largest group of resistance systems functions by energy-dependent efflux of toxic ions. Fewer involve enzymatic transformations (oxidation, reduction, methylation, and demethylation) or metal-binding proteins (for example, metallothionein SmtA, chaperone CopZ and periplasmic silver binding protein SilE). Some of the efflux resistance systems are ATPases and others are chemiosmotic ion/proton exchangers. For example, Cd2+-efflux pumps of bacteria are either inner membrane P-type ATPases or three polypeptide RND chemiosmotic complexes consisting of an inner membrane pump, a periplasmic-bridging protein and an outer membrane channel. In addition to the best studied three-polypeptide chemiosmotic system, Czc (Cd2+, Zn2+, and Co2), others are known that efflux Ag+, Cu+, Ni2+, and Zn2+. Resistance to inorganic mercury, Hg2+ (and to organomercurials, such as CH3Hg+ and phenylmercury) involve a series of metal-binding and membrane transport proteins as well as the enzymes mercuric reductase and organomercurial lyase, which overall convert more toxic to less toxic forms. Arsenic resistance and metabolizing systems occur in three patterns, the widely-found ars operon that is present in most bacterial genomes and many plasmids, the more recently recognized arr genes for the periplasmic arsenate reductase that functions in anaerobic respiration as a terminal electron acceptor, and the aso genes for the periplasmic arsenite oxidase that functions as an initial electron donor in aerobic resistance to arsenite.

  12. Acute toxicity bioassays of mercuric chloride and malathion on air-breathing fish Channa punctatus (Bloch).

    PubMed

    Pandey, Sanjay; Kumar, Ravindra; Sharma, Shilpi; Nagpure, N S; Srivastava, Satish K; Verma, M S

    2005-05-01

    Acute toxicity tests (96 h) were conducted in flow-through systems to determine the lethal toxicity of a heavy metal compound, mercuric chloride, and an organophosphorus pesticide, malathion, to air-breathing teleost fish, Channa punctatus (Bloch) and to study their behavior. The 96-h LC50 values were determined, as well as safe levels. The results indicate that mercuric chloride is more toxic than malathion to the fish species under study. Dose- and dose-time-dependent increases in mortality rate were also observed in response to both test chemicals.

  13. Mesozoic hydrothermal alteration associated with gold mineralization in the Mercur district, Utah

    SciTech Connect

    Wilson, P.N.; Parry, W.T. )

    1990-09-01

    K/Ar dates and chemical data show that a Mesozoic gold-bearing hydrothermal system altered black shales of the Mississippian Great Blue Limestone throughout an area encompassing the Mercur gold district, Utah. K/Ar dates of illite veins and illite-rich, clay-sized separates of altered shales that are enriched in Au, As, Hg, Sc, and other heavy metals indicate that hydrothermal activity occurred from 193 to 122 Ma. Several ages from within the Mercur district cluster near 160 Ma and may date the minimum age of gold mineralization.

  14. Comparative toxicogenomic responses of mercuric and methyl-mercury.

    PubMed

    McElwee, Matthew K; Ho, Lindsey A; Chou, Jeff W; Smith, Marjolein V; Freedman, Jonathan H

    2013-10-11

    Mercury is a ubiquitous environmental toxicant that exists in multiple chemical forms. A paucity of information exists regarding the differences or similarities by which different mercurials act at the molecular level. Transcriptomes of mixed-stage C. elegans following equitoxic sub-, low- and high-toxicity exposures to inorganic mercuric chloride (HgCl2) and organic methylmercury chloride (MeHgCl) were analyzed. In C. elegans, the mercurials had highly different effects on transcription, with MeHgCl affecting the expression of significantly more genes than HgCl2. Bioinformatics analysis indicated that inorganic and organic mercurials affected different biological processes. RNAi identified 18 genes that were important in C. elegans response to mercurial exposure, although only two of these genes responded to both mercurials. To determine if the responses observed in C. elegans were evolutionarily conserved, the two mercurials were investigated in human neuroblastoma (SK-N-SH), hepatocellular carcinoma (HepG2) and embryonic kidney (HEK293) cells. The human homologs of the affected C. elegans genes were then used to test the effects on gene expression and cell viability after using siRNA during HgCl2 and MeHgCl exposure. As was observed with C. elegans, exposure to the HgCl2 and MeHgCl had different effects on gene expression, and different genes were important in the cellular response to the two mercurials. These results suggest that, contrary to previous reports, inorganic and organic mercurials have different mechanisms of toxicity. The two mercurials induced disparate effects on gene expression, and different genes were important in protecting the organism from mercurial toxicity.

  15. Vapor growth of mercuric iodide tetragonal prismatic crystals

    NASA Astrophysics Data System (ADS)

    Ariesanti, Elsa

    The effect of polyethylene addition on the growth of mercuric iodide (HgI2) tetragonal prismatic crystals is examined. Three types of polyethylene powder are utilized: low molecular weight (Mw ˜ 4 x 103), ultra high molecular weight (Mw ˜ 3-6 x 1066), and spectrophotometric grade polyethylenes. Among these types of polyethylene, the low molecular weight polyethylene produces the most significant change in HgI2 morphology, with {110} being the most prominent crystal faces. Thermal desorption - gas chromatography/ mass spectroscopy (TD-GC/MS) studies show that thermal desorption of the low molecular weight polyethylene at 100°C and 150°C produce isomers of alkynes, odd nalkanes, and methyl (even-n) alkyl ketones. HgI2 growth runs with n-alkanes, with either neicosane, n-tetracosane, or n-hexatriacontane, cannot replicate the crystal shapes produced during growth with the low molecular weight polyethylene, whereas HgI2 growth runs with ketones, with either 3-hexadecanone or 14-heptacosanone, produce HgI2 tetragonal prismatic crystals, similar to the crystals grown with the low molecular weight polyethylene. C-O double bond contained in any ketone is a polar bond and this polar bond may be attracted to the mercury atoms on the top-most layer of the {110} faces through dipoledipole interaction. As a result, the growth of the {110} faces is impeded, with the crystals elongated in the [001] direction and bounded by the {001} faces along with large, prismatic {110} faces.

  16. Novel semiconductor radiation detector based on mercurous halides

    NASA Astrophysics Data System (ADS)

    Chen, Henry; Kim, Joo-Soo; Amarasinghe, Proyanthi; Palosz, Withold; Jin, Feng; Trivedi, Sudhir; Burger, Arnold; Marsh, Jarrod C.; Litz, Marc S.; Wiejewarnasuriya, Priyalal S.; Gupta, Neelam; Jensen, Janet; Jensen, James

    2015-08-01

    The three most important desirable features in the search for room temperature semiconductor detector (RTSD) candidate as an alternative material to current commercially off-the-shelf (COTS) material for gamma and/or thermal neutron detection are: low cost, high performance and long term stability. This is especially important for pager form application in homeland security. Despite years of research, no RTSD candidate so far can satisfy the above 3 features simultaneously. In this work, we show that mercurous halide materials Hg2X2 (X= I, Cl, Br) is a new class of innovative compound semiconductors that is capable of delivering breakthrough advances to COTS radiation detector materials. These materials are much easier to grow thicker and larger volume crystals. They can detect gamma and potentially neutron radiation making it possible to detect two types of radiation with just one crystal material. The materials have wider bandgaps (compared to COTS) meaning higher resistivity and lower leakage current, making this new technology more compatible with available microelectronics. The materials also have higher atomic number and density leading to higher stopping power and better detector sensitivity/efficiency. They are not hazardous so there are no environmental and health concerns during manufacturing and are more stable making them more practical for commercial deployment. Focus will be on Hg2I2. Material characterization and detector performance will be presented and discussed. Initial results show that an energy resolution better than 2% @ 59.6 keV gamma from Am-241 and near 1% @ 662 keV from Cs-137 source can be achieved at room temperature.

  17. Comparative toxicogenomic responses of mercuric and methyl-mercury

    PubMed Central

    2013-01-01

    Background Mercury is a ubiquitous environmental toxicant that exists in multiple chemical forms. A paucity of information exists regarding the differences or similarities by which different mercurials act at the molecular level. Results Transcriptomes of mixed-stage C. elegans following equitoxic sub-, low- and high-toxicity exposures to inorganic mercuric chloride (HgCl2) and organic methylmercury chloride (MeHgCl) were analyzed. In C. elegans, the mercurials had highly different effects on transcription, with MeHgCl affecting the expression of significantly more genes than HgCl2. Bioinformatics analysis indicated that inorganic and organic mercurials affected different biological processes. RNAi identified 18 genes that were important in C. elegans response to mercurial exposure, although only two of these genes responded to both mercurials. To determine if the responses observed in C. elegans were evolutionarily conserved, the two mercurials were investigated in human neuroblastoma (SK-N-SH), hepatocellular carcinoma (HepG2) and embryonic kidney (HEK293) cells. The human homologs of the affected C. elegans genes were then used to test the effects on gene expression and cell viability after using siRNA during HgCl2 and MeHgCl exposure. As was observed with C. elegans, exposure to the HgCl2 and MeHgCl had different effects on gene expression, and different genes were important in the cellular response to the two mercurials. Conclusions These results suggest that, contrary to previous reports, inorganic and organic mercurials have different mechanisms of toxicity. The two mercurials induced disparate effects on gene expression, and different genes were important in protecting the organism from mercurial toxicity. PMID:24118919

  18. Defects in red mercuric iodide related to device applications

    SciTech Connect

    Bao Xue Jun.

    1991-01-01

    Red mercuric iodide ({alpha}-HgI{sub 2}) is very promising material for fabrication of X-ray and {gamma}-ray detectors. Compared with conventional semiconductor nuclear detectors such as Si(Li), Ge(Li) and HPGe, HgI{sub 2} has a wider band gap (2.1 eV at room temperature), and higher atomic numbers. The advantages of HgI{sub 2} nuclear detectors as a consequence of these two properties are room temperature operation and high stopping power of X-ray or {gamma}-ray radiation. One major obstacle in taking full advantage of the potential of this material has been low manufacturing yield of high quality detectors. Both crystal growth and device fabrication are responsible for the introduction of defects in HgI{sub 2} crystals which lower the quality of the detectors. The author has employed several experimental techniques such as low temperature photoluminescence spectroscopy, thermally stimulated current measurements, and photoresponse measurements to study the incorporation of defects in the process of device fabrication. The effects of chemical etching, annealing, exposure to vacuum, surface heating, aging, and treatment with mercury and iodine were separately investigated. Interaction between various contact materials and HgI{sub 2} substrates, impurity identification using features in photoluminescence spectra, and the suitability of contact materials other than those presently being used for detector applications were also investigated. He has also found correlations between features in the photoluminescence spectra and the ability of HgI{sub 2} crystals to produce high quality detectors. With these correlations and understandings obtained by the studies of processing and contact deposition, suggestions were made to improve the fabrication procedures of HgI{sub 2} detectors. Finally, during the course of the study, he has also gained knowledge on the optical properties of HgI{sub 2}, which, at the moment, is very poorly understood.

  19. Discrete scintillator coupled mercuric iodide photodetector arrays for breast imaging

    SciTech Connect

    Tornai, M.P.; Levin, C.S.; Hoffman, E.J.

    1996-12-31

    Multi-element (4x4) imaging arrays with high resolution collimators, size matched to discrete CsI(Tl) scintillator arrays and mercuric iodide photodetector arrays (HgI{sub 2} PDA) are under development as prototypes for larger 16 x 16 element arrays. The compact nature of the arrays allows detector positioning in proximity to the breast to eliminate activity not in the line-of-sight of the collimator, thus reducing image background. Short collimators, size matched to {le}1.5 x 1.5 mm{sup 2} scintillators show a factor of 2 and 3.4 improvement in spatial resolution and efficiency, respectively, compared to high resolution collimated gamma cameras for the anticipated compressed breast geometries. Monte Carlo simulations, confirmed by measurements, demonstrated that scintillator length played a greater role in efficiency and photofraction for 140 keV gammas than cross sectional area, which affects intrinsic spatial resolution. Simulations also demonstrated that an increase in the ratio of scintillator area to length corresponds to an improvement in light collection. Electronic noise was below 40 e{sup -} RMS indicating that detector resolution was not noise limited. The high quantum efficiency and spectral match of prototype unity gain HgI{sub 2} PDAs coupled to 1 x 1 x 2.5 mm{sup 3} and 2 x 2 x 4 mm{sup 3} CsI(Tl) scintillators demonstrated energy resolutions of 9.4% and 8.8% FWHM at 140 keV, respectively, without the spectral tailing observed in standard high-Z, compound semi-conductor detectors. Line spread function measurements matched the scintillator size and pitch, and small, complex phantoms were easily imaged.

  20. Advanced mercuric iodide detectors for X-ray microanalysis

    SciTech Connect

    Warburton, W.K.; Iwanczyk, J.S.

    1987-01-01

    We first present a brief tutorial on Mercuric Iodide (HgI/sub 2/) detectors and the intimately related topic of near-room temperature ultralow noise preamplifiers. This provides both a physical basis and technological perspective for the topics to follow. We next describe recent advances in HgI/sub 2/ applications to x-ray microanalysis, including a space probe Scanning Electron Microscope (SEM), Synchrotron x-ray detectors, and energy dispersive detector arrays. As a result of this work, individual detectors can now operate stably for long periods in vacuum, detect soft x-rays to the oxygen K edge at 523 eV, or count at rates exceeding 2x10(5)/sec. The detector packages are small, lightweight, and use low power. Preliminary HgI/sub 2/ detector arrays of 10 elements with 500eV resolution have also been constructed and operate stably. Finally, we discuss expected advances in HgI/sub 2/ array technology, including improved resolution, vacuum operation, and the development of soft x-ray transparent encapsulants. Array capabilities include: large active areas, high (parallel) count rate capability and spatial sensitivity. We then consider areas of x-ray microanalysis where the application of such arrays would be advantageous, particularly including elemental microanalysis, via x-ray fluorescence spectroscopy, in both SEMs and in scanning x-ray microscopes. The necessity of high count rate capability as spatial resolution increases is given particular attention in this connection. Finally, we consider the possibility of Extended X-ray Absorption Fine Structure (EXAFS) studies on square micron sized areas, using detector arrays.

  1. De-mercurization of wastewater by Bacillus cereus (JUBT1): growth kinetics, biofilm reactor study and field emission scanning electron microscopic analysis.

    PubMed

    Ghoshal, Sanjukta; Bhattacharya, Pinaki; Chowdhury, Ranjana

    2011-10-30

    Removal of mercuric ions by a mercury resistant bacteria, called Bacillus cereus (JUBT1), isolated from the sludge of a local chlor-alkali industry, has been investigated. Growth kinetics of the bacteria have been determined. A multiplicative, non-competitive relationship between sucrose and mercury ions has been observed with respect to bacterial growth. A combination of biofilm reactor, using attached growth of Bacillus cereus (JUBT1) on rice husk packing, and an activated carbon filter has been able to ensure the removal of mercury up to near-zero level. Energy dispersive spectrometry analysis of biofilm and the activated carbon has proved the transformation of Hg(2+) to Hg(0) and its confinement in the system. Copyright © 2011 Elsevier B.V. All rights reserved.

  2. Scrubbing of iodine from gas streams with mercuric nitrate-conversion of mercuric iodate product to barium iodate for fixation in concrete

    SciTech Connect

    Rogers, G.C.; Moore, J.G.; Morgan, M.T.

    1980-06-01

    A bench-scale model of a mercuric nitrate scrubber for removal of iodine from off-gas streams was constructed and operated in conjunction with a mercuric iodate-to-barium iodate conversion system to determine the feasibility of total recycle of all processing solutions. The two main aspects of the system examined were (1) the extent of contamination of the barium iodate product, and (2) the effect of cross-contamination of various process solutions on the efficiency of the process. The experimental evidence obtained indicates that, with appropriate control, all solutions can be recycled without significant contamination of the product that would be harmful to the host concrete or to the environment. Mercury contamination was found to be less than or equal to 0.5 wt % of the barium iodate product. The most significant effect on system efficiency was determined to be barium hydroxide contamination of the sodium hydroxide solution used to convert mercuric iodate to sodium iodate. A mole ratio of barium hydroxide to sodium hydroxide of about 1:225 caused a decrease in conversion efficiency of about 45%.

  3. Electrical properties of solid iodo mercurates resulting from the reaction of HgI2 with alcaline iodides

    NASA Astrophysics Data System (ADS)

    Ponpon, J. P.; Amann, M.

    2005-01-01

    Potassium iodide solutions are currently used during the fabrication process of mercuric iodide based nuclear radiation detectors. However, KI treatment leaves the HgI2 surface covered with a residual compound (namely the potassium tri-iodo mercurate) which has a significant influence on the surface properties and stability of mercuric iodide devices and therefore on the detectors characteristics. Looking for other solutions to etch mercuric iodide, we found it interesting to investigate the electrical properties of the compounds which may form when etching HgI2 in NH4I, NaI, and RbI. For this purpose, solid iodo mercurates with the cations ammonium, sodium, and rubidium, have been prepared by reacting HgI2 with the solutions of interest. Study of the electrical properties of these samples and comparison with those of potassium tri-iodo mercurate ones, especially with respect to humidity, indicates noticeable stability differences in presence of water vapour. This could have interesting consequences on the surface cleaning of mercuric iodide.

  4. Ultrastructural alterations produced in cockerels after mercuric chloride toxicity and subsequent interaction with an organophosphate insecticide.

    PubMed

    Chishti, M A; Rotkiewicz, T

    1992-05-01

    Ultrastructural changes of liver and kidneys of cockerels exposed to mercuric chloride and subsequent interaction with methylobromofenvinphos (IPO 63 compound) were studied. Group A birds were treated for 4 weeks with 300 ppm mercuric chloride in drinking water; Group B birds were treated for 4 weeks with mercuric chloride followed by single oral dose of 240 mg/kg of IPO 63; Group C 240 mg/kg IPO 63 only; and Group D, unexposed controls. Hepatocytes of mercury-IPO 63 interaction group B showed large lysosomes containing myelin bodies, swollen mitochondria with cristeolysis, dilated endoplasmic reticulum and numerous vacuoles containing granular material. Mercury-intoxicated birds showed similar but less severe changes, whereas IPO 63-treated birds showed accumulation of glycogen granules, fat droplets, and few lysosomal bodies as well as other changes. Renal corpuscles of kidney from mercury-IPO 63 interaction birds revealed minor ultrastructural changes as vacuolation, swollen mitochondria of podocytes and slight thickening of the glomerular basement membrane. Proximal tubular cells showed extreme damage such as, microvillar loss, dilation of endoplasmic reticulum, accumulation of lysosomal bodies, glycogen granules, myelin figures, swollen mitochondria with granular material, numerous vacuoles containing degenerated membranous organelles and distorted, pyknotic nucleus with marked dilation of nuclear membrane. Mercury intoxicated birds showed similar but less pronounced changes in tubules. These observations suggest that the effect of mercuric chloride toxicity and then interaction with an organophosphorus insecticide causes extreme damage to hepatic and renal cells that appears to be additive.

  5. Sexual maturation and productivity of Japanese quail fed graded concentrations of mercuric chloride.

    PubMed

    Hill, E F; Shaffner, C S

    1976-07-01

    Japanese quail (Coturnix c. japonica) were fed 0, 2, 4, 8, 16, and 32 p.p.m. Hg as mercuric chloride (HgCl2) from the time of hatching up to the age of 1 year. None of the birds manifested any gross signs of mercury poisioning. Food consumption, growth rate, and weight maintenance were unaffected. Initial oviposition tended to occur at a younger age as dietary mercuric chloride increased, e.g., the median age at which egg laying began among hens fed 32 p.p.m. Hg was 6 days younger than for controls. The average rate of egg production was positively related to the concentration of mercuric chloride with the most pronounced differences between treatments occurring among young (less than 9-week-old) hens. Beyond 9 weeks of age production was more uniform among the treatments, but even after 1 year hens on 32 p.p.m. Hg were laying an average of 13.5% more eggs than controls. Rate of egg fertilization was generally depressed for all Hg-treatments above 4 p.p.m. Hatchability of fertilized eggs and eggshell thickness appeared unaffected by mercuric chloride.

  6. The use of a mercuric iodide detector for X-ray fluorescence analysis in archaeometry

    NASA Astrophysics Data System (ADS)

    Cesareo, R.; Gigante, G. E.; Iwanczyk, J. S.; Dabrowski, A.

    1992-11-01

    For about two decades, energy dispersive X-ray fluorescence (EDXRF) has been employed in Rome for the analysis of works of art. A short history of the applications of EDXRF to paintings and alloys is presented. Finally, the usefulness of mercuric iodide room-temperature semiconductor detectors in this field is shown.

  7. Sexual maturation and productivity of Japanese quail fed graded concentrations of mercuric chloride

    USGS Publications Warehouse

    Hill, E.F.; Shaffner, C.S.

    1976-01-01

    Japanese quail (Coturnix c. japonica) were fed 0, 2, 4, 8, 16, and 32 p.p.m. Hg as mercuric chloride (HgCl2) from the time of hatching up to the age of 1 year. None of the birds manifested any gross signs of mercury poisioning. Food consumption, growth rate, and weight maintenance were unaffected. Initial oviposition tended to occur at a younger age as dietary mercuric chloride increased, e.g., the median age at which egg laying began among hens fed 32 p.p.m. Hg was 6 days younger than for controls. The average rate of egg production was positively related to the concentration of mercuric chloride with the most pronounced differences between treatments occurring among young (less than 9-week-old) hens. Beyond 9 weeks of age production was more uniform among the treatments, but even after 1 year hens on 32 p.p.m. Hg were laying an average of 13.5% more eggs than controls. Rate of egg fertilization was generally depressed for all Hg-treatments above 4 p.p.m. Hatchability of fertilized eggs and eggshell thickness appeared unaffected by mercuric chloride.

  8. Metal oxide and mercuric sulfide nanoparticles synthesis and characterization

    NASA Astrophysics Data System (ADS)

    Xu, Xin

    Commercially available and laboratory-synthesized metal based nanoparticles (NPs), iron oxide (Fe2O3), copper oxide (CuO), titanium dioxide (TiO2), zinc oxide (ZnO) and mercuric sulfide (HgS) were studied by comprehensive characterizations methods. The general synthesis process was modified sol-gel method. The size and morphology of NPs could be influenced by temperature, sonication, calcination, precursor concentration, pH and types of reaction media. All types of the laboratory-synthesized or commercially available NPs were characterized by physical and chemical processes. One characteristic of NP that can lead to ambiguous toxicity test results was the effect of agglomeration of primary nano-sized particles. Laser light scattering was used to measure the aggregated and particle size distribution. Aggregation effects were apparent and often extensive in some synthesis approaches. Electron microscopy (SEM and TEM) gave the images of those laboratory-synthesized particles and aggregation. The average single particle was about 5-20 nm of ZnO; 20-40 nm of CuO; 10-20 nm of TiO2; 20-35 nm of Fe2O3; 10-15 nm of HgS, while the aggregate size was in the range of a hundred nanometers or more. These five types of NPs were obtained with spherical and oblong formation and the agglomeration of ZnO, CuO, HgS and TiO2 was random, but Fe2O3 has web-like aggregation. Other measurements performed on the particles and aggregates include bandgap energies, surface composition, surface area, hydrodynamic radius, and particle surface charge. In aqueous environment, NPs are subject to processes such as solubilization and aggregation. These processes can be controlling factors in the fate of nanomaterials in environmental settings, including bioavailability to organisms. This study has focused primarily on measurement of the solubility in aqueous media of varying composition (pH, ionic strength, and organic carbon), sedimentation and stability. The aggregate size distribution was

  9. Renal accumulation and intrarenal distribution of inorganic mercury in the rabbit: Effect of unilateral nephrectomy and dose of mercuric chloride

    SciTech Connect

    Zalups, R.K. )

    1991-06-01

    The effects of unilateral nephrectomy and dose of mercuric chloride on the short-term renal accumulation and intrarenal distribution of inorganic mercury were studied in the rabbit. The renal accumulation of inorganic mercury, on a per gram basis, was increased in uninephrectomized (NPX) rabbits compared with that in sham-operated (SO) rabbits 24 h after the animals received either a nontoxic 2.0 mumol/kg or nephrotoxic 4.0 mumol/kg dose of mercuric chloride. In the NPX rabbits given the 2.0 mumol/kg dose of mercuric chloride, the increased accumulation of inorganic mercury was due to increased accumulation of mercury in the outer stripe of the outer medulla. In the NPX rabbits given the 4.0 mumol/kg dose of mercuric chloride, the increased renal accumulation of mercury appeared to be due to increased accumulation of mercury in both the renal cortex and outer stripe of the outer medulla. Interestingly, no differences in the renal accumulation of inorganic mercury were found between NPX and SO rabbits given a low nontoxic 0.5 mumol/kg dose of mercuric chloride. As the dose of mercuric chloride was increased from 0.5 to 4.0 mumol/kg, the percent of the administered dose of mercury that accumulated in each gram of renal tissue decreased substantially. The findings in the present study indicate that the renal accumulation of inorganic mercury increases after unilateral nephrectomy when certain nontoxic and nephrotoxic doses of mercuric chloride are administered. In addition, they indicate that the percent of the administered dose of mercury that accumulates in the renal tissue of both NPX and SO rabbits decreases as the dose of mercuric chloride is increased.

  10. Protein expression of kidney and liver bilitranslocase in rats exposed to mercuric chloride--a potential tissular biomarker of toxicity.

    PubMed

    Trebucobich, Mara Soledad; Hazelhoff, María Herminia; Chevalier, Alberto A; Passamonti, Sabina; Brandoni, Anabel; Torres, Adriana Mónica

    2014-03-03

    Bilitranslocase (BTL) is a plasma membrane carrier that transports organic anions of physiological and pharmacological interest. It is expressed in basolateral plasma membrane of kidney and liver. BTL has been recently described as a marker of transition from normal tissue to its neoplastic transformation in human kidney. Inorganic mercury is a major environmental contaminant that produces many toxic effects. Previous reports have described an interaction between BTL and mercuric ions. This study was designed to evaluate the renal and hepatic expression of BTL in rats exposed to a nephrotoxic and hepatotoxic dose of HgCl2. Male rats were treated with a single injection of HgCl2 at a dose of 4mg/kg body wt, i.p. (HgCl2 group). Control rats received the vehicle alone (Control group). Studies were carried out 18h after injection. Afterwards, the kidneys and livers were excised and processed for histopathological studies or immunoblot (homogenates and crude membranes) techniques. In rats treated with HgCl2, immunoblotting showed a significant decrease in the abundance of BTL in homogenates and plasma membranes from kidney and liver. BTL decrease of expression might reflect the grade of damage in renal tubule cells and in hepatocytes. Thus, BTL might be postulated as a new biomarker of tissue toxicity induced by mercury.

  11. Mercuric chloride-induced testicular toxicity in rats and the protective role of sodium selenite and vitamin E.

    PubMed

    Kalender, Suna; Uzun, Fatma Gokce; Demir, Filiz; Uzunhisarcıklı, Meltem; Aslanturk, Ayse

    2013-05-01

    Mercury has been recognized as an environmental pollutant that adversely affects male reproductive systems of animals. This study examined the effects of mercuric chloride on the antioxidant system and histopathological changes and also evaluated the ameliorating effects of sodium selenite and/or vitamin E in the rat testis tissues. Sexually mature male Wistar rats (weighing 300-320g and each group six animals) were given mercuric chloride (1mg/kg bw) and/or sodium selenite (0.25mg/kg bw)+vitamin E (100mg/kg) daily via gavage for 4weeks. In the present study, mercuric chloride exposure resulted in an increase in the TBARS level and a decrease in the SOD, CAT, GPx activities, with respect to the control. Further, light microscopic investigation revealed that mercury exposure induced histopathological alterations in the testis tissues. Supplementation of sodium selenite and/or vitamin E to mercury-induced groups declined lipid peroxidation, increased SOD, CAT, GPx activities. While some histopathological changes were detected in mercuric chloride treated group, milder histopathological changes were observed in animal co-treated with sodium selenite and/or vitamin E supplementation to mercuric chloride-treated rats. As a result, mercuric chloride induced testicular toxicity is reduced by sodium selenite and/or vitamin E, but not ameliorate completely.

  12. Construction and evaluation of a metal ion biosensor

    NASA Astrophysics Data System (ADS)

    Tescione, Lia; Belfort, Georges

    1993-01-01

    E. coli, genetically engineered with a mercury(II)-sensitive promoter and the lux genes from Vibrio fischeri, were used as microbial sensors for the detection of mercury. Evaluation of this genetic construction was carried out by determining the effects of various parameters on cell suspensions maintained at constant conditions in a small vessel. The strongest light intensities and quickest induction times occurred with cells in the mid-exponential growth phase maintained at 280 C, concentrated to 1 x 10(exp 9) cells/mL, mixed at very fast speeds, and aerated at 2 vvm (volume of air per volume of culture per minute) during light measurement in the small vessel. The sensitivity of these cells to the mercuric ion lied in the range of 0.02-4 micrometer (4-800 ppb) and the total response time was on the order of one hour, depending on the above parameters. The cells exhibited great specificity for mercury. The cells have almost equal specificity for organic and inorganic form of the mercuric ion and responded more weakly to the mercurous ion. A simple, inexpensive, durable miniature probe was constructed and operated using the optimum parameters found in the small vessel as a guide. The range of sensitivity to the mercuric ion detected in the probe was 0.01-4 micrometer when aeration was provided.

  13. Construction and evaluation of a metal ion biosensor

    SciTech Connect

    Tescione, L.; Belfort, G. . Dept. of Chemical Engineering)

    1993-10-01

    Escherichia coli, genetically engineered with a mercury (II)-sensitive promoter and the lux genes from Vibrio fischeri, were used as microbial bioluminescent sensors for the detection of mercury. Evaluation of this genetic construction was carried out by determining the effects of various parameters on cell suspensions maintained at constant conditions in a small 100-mL vessel. The strongest light intensities and quickest induction times occurred with cells in the midexponential growth phase maintained at 28C, concentrated to 1 [times] 10[sup 9] cells/mL, mixed at very fast speeds, and aerated at 2 vvm during light measurement in the small vessel. The cells were sensitive to the mercuric ion in the range of 20 nM to 4 [mu]M, and the total response time was on the order of 1 hour, depending on the above parameters. The cells exhibited great specificity for mercury. The cells had almost equal specificity for organic and inorganic forms of the mercuric ion and responded more weakly to the mercurous ion. A simple, inexpensive, durable miniature probe was constructed and operated using the optimum parameters found in the small vessel as a guide. The range of sensitivity to the mercuric ion detected in the probe was 10 nM to 4 [mu]M when aeration was provided.

  14. Characterization of thyroidal glutathione reductase

    SciTech Connect

    Raasch, R.J.

    1989-01-01

    Glutathione levels were determined in bovine and rat thyroid tissue by enzymatic conjugation with 1-chloro-2,4-dinitrobenzene using glutathione S-transferase. Bovine thyroid tissue contained 1.31 {+-} 0.04 mM reduced glutathione (GSH) and 0.14 {+-} 0.02 mM oxidized glutathione (GSSG). In the rat, the concentration of GSH was 2.50 {+-} 0.05 mM while GSSG was 0.21 {+-} 0.03 mM. Glutathione reductase (GR) was purified from bovine thyroid to electrophoretic homogeneity by ion exchange, affinity and molecular exclusion chromatography. A molecular weight range of 102-109 kDa and subunit size of 55 kDa were determined for GR. Thyroidal GR was shown to be a favoprotein with one FAD per subunit. The Michaelis constants of bovine thyroidal GR were determined to be 21.8 {mu}M for NADPH and 58.8 {mu}M for GSSG. The effect of thyroid stimulating hormone (TSH) and thyroxine (T{sub 4}) on in vivo levels of GR and glucose 6-phosphate dehydrogenase were determined in rat thyroid homogenates. Both enzymes were stimulated by TSH treatment and markedly reduced following T{sub 4} treatment. Lysosomal hydrolysis of ({sup 125}I)-labeled and unlabeled thyroglobulin was examined using size exclusion HPLC.

  15. Role of Tribulus terrestris (Linn.) (Zygophyllacea) against mercuric chloride induced nephrotoxicity in mice, Mus musculus (Linn.).

    PubMed

    Kavitha, A V; Jagadeesan, G

    2006-05-01

    The present study investigates the influence of methanolic fraction (MF) of Tribulus terrestris fruit extract on the kidney tissues of mercury intoxicated mice, Mus musculus. At median-lethal dose of mercuric chloride (12.9 mg/kg body weight), the whole kidney tissue showed an increased level of lipid peroxidation (LPO), glutathione peroxidase (GPx) activities and simultaneously a decreased level of superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH). These parameters reached to near normal after administration of fruit extracts of T. terrestris for 7 days. The results suggested that the oral administration of methanolic fraction of Tribulus terrestris fruit extract at dose 6 mg/kg body weight provided protection against the mercuric chloride induced toxicity in the mice.

  16. Growth of mercuric iodide (HgI2) for nuclear radiation detectors

    NASA Technical Reports Server (NTRS)

    Vandenberg, L.; Schnepple, W. F.

    1988-01-01

    Mercuric iodide is a material used for the fabrication of the sensing element in solid state X-ray and gamma ray detecting instruments. The operation of the devices is determined to a large degree by the density of structural defects in the single crystalline material used in the sensing element. Since there were strong indications that the quality of the material was degraded by the effects of gravity during the growth process, a research and engineering program was initiated to grow one or more crystals of mercuric iodide in the reduced gravity environment of space. A special furnace assembly was designed which could be accommodated in a Spacelab rack, and at the same time made it possible to use the same growth procedures and controls used when growing a crystal on the ground. The space crystal, after the flight, was subjected to the same evaluation methods used for earth-grown crystals, so that comparisons could be made.

  17. Acute effects of mercuric chloride on glycogen and protein content of zebra fish, Danio rerio.

    PubMed

    Vutukuru, S S; Basani, Kalpana

    2013-03-01

    Presence of mercury and other heavy metals above permissible levels in water bodies across the globe is posing a serious threat to aquatic biota and public health. Occurrence of mercury above the permissible limits in the aquatic ecosystem of Hyderabad city is well established. In this context, we carried out static- renewal bioassays on the zebra fish, Danio rerio exposed to different concentrations of mercuric chloride, and the 96-h median lethal concentration (LC50) was found to be 0.077 mgl(-1). Behavioral manifestations like loss of scales, hyper secretion of mucus, surfacing and darting movements, loss of balance, irregular swimming patterns were noticed in the fish exposed to 0.077 mgl(-1). The present study also examined the toxic effects of mercuric chloride on vital biochemical constituent's total glycogen and total protein. Significant decrease (p < 0.001) in glycogen and protein content of fish exposed to 0.077 mgl(-1).

  18. Impairment of mitochondrial energy metabolism of two marine fish by in vitro mercuric chloride exposure.

    PubMed

    Mieiro, C L; Pardal, M; Duarte, A; Pereira, E; Palmeira, C M

    2015-08-15

    The goal of this work was to understand the extent of mercury toxic effects in liver metabolism under an episode of acute contamination. Hence, the effects of in vitro mercuric chloride in liver mitochondria were assessed in two commercial marine fish: Senegalese sole (Solea senegalensis) and gilthead seabream (Sparus aurata). Liver mitochondria were exposed to 0.2mgL(-1) of mercury, the average concentration found in fish inhabiting contaminated areas. Mercuric chloride depressed mitochondrial respiration state 3 and the maximal oxygen consumption in the presence of FCCP indicating inhibitory effects on the oxidative phosphorylation and on the electron transport chain, respectively. The inhibition of F1Fo-ATPase and succinate-dehydrogenase activities also corroborated the ability of mercury to inhibit ADP phosphorylation and the electron transport chain. This study brings new understanding on the mercury levels able to impair fish mitochondrial function, reinforcing the need for further assessing bioenergetics as a proxy for fish health status.

  19. A study of the homogeneity and deviations from stoichiometry in mercuric iodide

    NASA Astrophysics Data System (ADS)

    Burger, A.; Morgan, S.; He, C.; Silberman, E.; van den Berg, L.; Ortale, C.; Franks, L.; Schieber, M.

    1990-01-01

    We have been able to determine the deviations from stoichiometry of mercuric iodide (HgI 2) by using differential scanning calorimetry (DSC). Mercury excess or iodine deficiency in mercuric iodide can be evaluated from the eutectic melting of α-Hgl 2-Hg 2I 2 at 235 °C, which appears as an additional peak in DSC thermograms. I 2 excess can be found from the existence of the I 2-α-HgI 2 eutectic melting at 103°C. An additional DSC peak appears in some samples around 112°C, that could be explained by the presence of iodine inclusions. Using resonance fluorescence spectroscopy (RFS) we have been able to determine the presence of free I 2 that is released by samples during the heating at 120 °C (crystal growth temperature), thus giving additional support to the above DSC results.

  20. Effects of indium and tin overlayers on the photoluminescence spectrum of mercuric iodide

    SciTech Connect

    James, R.B. ); Bao, X.J.; Schlesinger, T.E. ); Ortale, C.; Cheng, A.Y. )

    1990-03-01

    Mercuric iodide (HgI{sub 2} ) crystals with semitransparent metal overlayers of indium and tin were characterized using low-temperature photoluminescence (PL) spectroscopy. The PL spectra were found to differ for points beneath the thin metal overlayers and points that were masked off during each deposition. The photoluminescence data were compared with PL measurements taken on HgI{sub 2} photodetectors with indium-tin-oxide (ITO) entrance electrodes. The similarities of the spectra for the HgI{sub 2} samples with In, Sn, and ITO conducting overlayers indicate that the regions in the ITO-contacted photodetectors with relatively poor photoresponses are associated with the interaction of indium or tin with the mercuric iodide substrate.

  1. Dose and sex dependent distribution of mercury in rats exposed to mercuric chloride

    SciTech Connect

    Khan, A.T.; Graham, T.C.; Webster, J.E.; Ferguson, J.A.

    1994-12-31

    A 14-day study was conducted in young male and female rats (Sprague-Dawley SDTM) with mercuric chloride at daily oral doses of 0, 1.25, 5.0, and 10.0 mg/kg mercuric chloride to determine the maximum tolerated dose and the distribution of mercury in the target organs. The brains, hearts, kidneys, livers, lungs and spleens of both male and female rats (survived or died during the experiment) were analyzed for mercury content. At all treatments (1.25, 2.5, 5.0, and 10.0 mg/kg) groups, mercury level was higher in the kidneys of both sexes, and followed by the livers, spleen, lungs, hearts, and brains, respectively. The mercury level in target organs of females was higher than those of males. All mercury treated rats also showed a reduction in cumulative body weight gained beginning on the third day of treatment.

  2. Use of mercuric iodide x-ray detectors with alpha backscattering spectrometers for space applications

    SciTech Connect

    Iwanczyk, J.S.; Wang, Y.J.; Dorri, N.; Dabrowski, A.J. ); Economou, T.E.; Turkevich, A.L. . Enrico Fermi Inst.)

    1991-04-01

    This paper presents x-ray fluorescence (XRF) spectra of different extraterrestrial samples taken with a mercuric iodide (HgI{sub 2}) spectrometer inserted into an Alpha Backscattering Instrument identical to that used in the Soviet Phobos Mission. The results obtained with the HgI{sub 2} ambient temperature detector are compared with those obtained using a Si(Li) cryogenically cooled detector. The authors' efforts to design an optimized instrument for space application are described.

  3. Effect of temperature gradient on the optical quality of mercurous chloride crystals

    NASA Technical Reports Server (NTRS)

    Singh, N. B.; Davies, D. K.; Gottlieb, M.; Henningsen, T.; Mazelsky, R.

    1989-01-01

    Single crystals of mercurous chloride were grown at temperature gradients of 8, 11 and 17 K/cm by the physical vapor transport method. The optical quality of these crystals was evaluated by measuring bulk scattering and inhomogeneity of refractive index by birefringence interferometry. It was observed that a high temperature gradient at the solid-vapor interface induced thermal stresses and crystals showed higher scattering and irregular fringes.

  4. Mercurated nucleotides: assessment of a new tool to study RNA synthesis and processing in isolated nuclei.

    PubMed Central

    Schäfer, K P

    1977-01-01

    Mercurated pyrimidine nucleotides have been used to study RNA synthesis and processing in isolated nuclei from mouse L cells. 5-mercuridine triphosphate (5-Hg-UTP) or 5-Hg-CTP are accepted as substrates by the purified RNA polymerases (I+III) and (II) from mouse cells, respectively, as well as by the enzymes still bound to the nuclear chromatin. In nuclei, RNA synthesis in the presence of Hg-UTP is reduced to 60-70% of a control. 30-60% of RNA labeled in vitro with (3H)UTP in isolated nuclei is not retained on sulfhydryl sepharose columns. Sucrose gradient analysis reveals a size distribution of the non-bound RNA similar to non-mercurated control RNA. Hg-RNA is found in a single peak from 4-10S. Chase experiments indicate that this RNA is the original transcript. It is argued that Hg-nucleotides may cause premature chain termination. Methylation of RNA in vitro by S-adenosyl methionine ((3H)SAM) is reduced to 75% of controls in the presence of Hg-UTP. Only 6% of the methyl groups appear in Hg-RNA. Polyadenylation is reduced as well. 15% of poly(A) (+)RNA are found in control assays whereas only 1% of Hg-RNA carries a poly(A) end added in vitro. These results limit the use of mercurated nucleotides for studies of nuclear RNA synthesis and processing. PMID:600804

  5. Hematotoxicity and genotoxicity of mercuric chloride following subchronic exposure through drinking water in male rats.

    PubMed

    Boujbiha, Mohamed Ali; Ben Salah, Ghada; Ben Feleh, Abdelraouf; Saoudi, Mongi; Kamoun, Hassen; Bousslema, Ali; Ommezzine, Asma; Said, Khaled; Fakhfakh, Faiza; El Feki, Abdelfattah

    2012-07-01

    Erythrocytes are a convenient model to understand the subsequent oxidative deterioration of biological macromolecules in metal toxicities. The present study examined the variation of hematoxic and genotoxic parameters following subchronic exposure of mercuric chloride via drinking water and their possible association with oxidative stress. Male rats were exposed to 50 ppm (HG1) and 100 ppm (HG2) of mercuric chloride daily for 90 days. A significant dose-dependent decrease was observed in red blood cell count, hemoglobin, hematocrit, and mean cell hemoglobin concentration in treated groups (HG1 and HG2) compared with controls. A significant dose-dependent increase was observed in lipid peroxidation; therefore, a significant variation was found in the antioxidant enzyme activities, such as superoxide dismutase, catalase, and glutathione peroxidase. Interestingly, mercuric chloride treatment showed a significant dose-dependent increase in frequency of total chromosomal aberration and in percentage of aberrant bone marrow metaphase of treated groups (p < 0.01). The oxidative stress induced by mercury treatment may be the major cause for chromosomal aberration as free radicals lead to DNA damage. These data will be useful in screening the antioxidant activities of natural products, which may be specific to the bone marrow tissue.

  6. Mercuric iodide detector systems for identifying substances by x-ray energy dispersive diffraction

    SciTech Connect

    Iwanczyk, J.S.; Patt, B.E.; Wang, Y.J.; Croft, M.; Kalman, Z.; Mayo, W.

    1995-08-01

    The use of mercuric iodide arrays for energy-dispersive x-ray diffraction (EDXRD) spectroscopy is now being investigated by the authors for inspection of specific crystalline powders in substances ranging from explosives to illicit drugs. Mercuric iodide has been identified as the leading candidate for replacing the Ge detectors previously employed in the development of this technique because HgI{sub 2} detectors: operate at or near room temperature; without the bulky apparatus associated with cryogenic cooling; and offer excellent spectroscopy performance with extremely high efficiency. Furthermore, they provide the practicality of constructing optimal array geometries necessary for these measurements. Proof of principle experiments have been performed using a single-HgI{sub 2} detector spectrometer. An energy resolution of 655 eV (FWHM) has been obtained for 60 keV gamma line from an {sup 241}Am source. The EDXRD signatures of various crystalline powdered compounds have been measured and the spectra obtained show the excellent potential of mercuric iodide for this application.

  7. Nitrate and periplasmic nitrate reductases

    PubMed Central

    Sparacino-Watkins, Courtney; Stolz, John F.; Basu, Partha

    2014-01-01

    The nitrate anion is a simple, abundant and relatively stable species, yet plays a significant role in global cycling of nitrogen, global climate change, and human health. Although it has been known for quite some time that nitrate is an important species environmentally, recent studies have identified potential medical applications. In this respect the nitrate anion remains an enigmatic species that promises to offer exciting science in years to come. Many bacteria readily reduce nitrate to nitrite via nitrate reductases. Classified into three distinct types – periplasmic nitrate reductase (Nap), respiratory nitrate reductase (Nar) and assimilatory nitrate reductase (Nas), they are defined by their cellular location, operon organization and active site structure. Of these, Nap proteins are the focus of this review. Despite similarities in the catalytic and spectroscopic properties Nap from different Proteobacteria are phylogenetically distinct. This review has two major sections: in the first section, nitrate in the nitrogen cycle and human health, taxonomy of nitrate reductases, assimilatory and dissimilatory nitrate reduction, cellular locations of nitrate reductases, structural and redox chemistry are discussed. The second section focuses on the features of periplasmic nitrate reductase where the catalytic subunit of the Nap and its kinetic properties, auxiliary Nap proteins, operon structure and phylogenetic relationships are discussed. PMID:24141308

  8. Regulation of Nitrate Reductase Activity in Corn (Zea mays L.) Seedlings by Endogenous Metabolites 1

    PubMed Central

    Schrader, L. E.; Hageman, R. H.

    1967-01-01

    Primary and secondary metabolites of inorganic nitrogen metabolism were evaluated as inhibitors of nitrate reductase (EC 1.6.6.1) induction in green leaf tissue of corn seedlings. Nitrite, nitropropionic acid, ammonium ions, and amino acids were not effective as inhibitors of nitrate reductase activity or synthesis. Increasing α-amino nitrogen and protein content of intact corn seedlings by culture techniques significantly enhanced rather than decreased the potential for induction of nitrate reductase activity in excised seedlings. Secondary metabolites, derived from phenylalanine and tyrosine, were tested as inhibitors of induction of nitrate reductase. Of the 9 different phenylpropanoid compounds tested, only coumarin, trans-cinnamic and trans-o-hydroxycinnamic acids inhibited induction of nitrate reductase. While coumarin alone exhibited a relatively greater inhibitory effect on enzyme induction than on general protein synthesis (the latter measured by incorporation of labeled amino acids), this differential effect may have been dependent upon unequal rates of synthesis and accumulation with respect to the initial levels of nitrate reductase and general proteins. Because of the short half-life of nitrate reductase, inhibitors of protein synthesis in general could still achieve differential regulation of nitrogen metabolism. Coumarin did not inhibit nitrate reductase activity when added directly to the assay mixture at 5 mm. Carbamyl phosphate and its chemical derivative, cyanate, were found to be competitive (with nitrate) inhibitors of nitrate reductase. The data suggest that cyanate is the active inhibitor in the carbamyl phosphate preparations. PMID:16656715

  9. A non-radioactive in situ hybridization method based on mercurated nucleic acid probes and sulfhydryl-hapten ligands.

    PubMed Central

    Hopman, A H; Wiegant, J; Tesser, G I; Van Duijn, P

    1986-01-01

    Mercurated nucleic acid probes can be used for non-radioactive in situ hybridization. The principle of the method is based on the reaction of the mercurated pyrimidine residues of the in situ hybridized probe with the sulfhydryl group of a ligand which contains a hapten. Next, the hapten is immunocytochemically detected. Previous experiments showed that stable coupling of the sulfhydryl ligands could only be obtained when positively charged amino groups are present in the ligand. On basis of this finding, ligands were synthesized containing a sulfhydryl group, two lysyl residues and hapten groups such as trinitrophenyl, fluorescyl and biotinyl. The ligands, free or bound to mercurated nucleic acids, were immunochemically characterized in ELISAs. The method was shown to be specific and sensitive in the detection of target DNA in situ on microscopic preparations and in dot-blot hybridization reactions on nitrocellulose. Images PMID:3748817

  10. Effects of mercuric chloride on (/sup 3/H)dopamine release from rat brain striatal synaptosomes

    SciTech Connect

    Hare, M.F.; Minnema, D.J.; Cooper, G.P.; Michaelson, I.A.

    1989-06-15

    Electrophysiological studies employing amphibian neuromuscular preparations have shown that mercuric chloride (HgCl2) in vitro increases both spontaneous and evoked neurotransmitter release. The present study examines the effect of HgCl2 on the release of (/sup 3/H)dopamine from synaptosomes prepared from mammalian brain tissue. Mercuric chloride (3-10 microM) produces a concentration-dependent increase in spontaneous (/sup 3/H)dopamine release from ''purified'' rat striatal synaptosomes, in both the presence and absence of extra-synaptosomal calcium. The effects of HgCl2 on transmitter release from amphibian neuromuscular junction preparations resemble those produced by the Na+, K+-ATPase inhibitor ouabain. Experiments were performed to determine whether the HgCl2 effects on mammalian synaptosomal dopamine release are a consequence of Na+, K+-ATPase inhibition. Na+, K+-ATPase activity in lysed synaptosomal membranes is inhibited by HgCl2 (IC50 = 160 nM). However, mercuric chloride in the presence of 1 mM ouabain still increased (3H)dopamine release. The specific inhibitor of Na+-dependent, high-affinity dopamine transport, RMI81,182 inhibited ouabain-induced (3H)dopamine release whereas it had no effect on HgCl2-induced (/sup 3/H)dopamine release. These data suggest that augmentation of spontaneous (/sup 3/H)dopamine release by HgCl2 probably is not mediated by an inhibition of Na+, K+-ATPase and HgCl2 does not act directly on the dopamine transporter.

  11. X-ray fluorescence analysis of alloy and stainless steels using a mercuric iodide detector

    NASA Technical Reports Server (NTRS)

    Kelliher, Warren C.; Maddox, W. Gene

    1988-01-01

    A mercuric iodide detector was used for the XRF analysis of a number of NBS standard steels, applying a specially developed correction method for interelemental effects. It is shown that, using this method and a good peak-deconvolution technique, the HgI2 detector is capable of achieving resolutions and count rates needed in the XRF anlysis of multielement samples. The freedom from cryogenic cooling and from power supplies necessary for an electrically cooled device makes this detector a very good candidate for a portable instrument.

  12. Transport phenomena during vapor growth of optoelectronic material - A mercurous chloride system

    NASA Technical Reports Server (NTRS)

    Singh, N. B.

    1990-01-01

    Crystal growth velocity was measured in a mercurous chloride system in a two-zone transparent furnace as a function of the Rayleigh number by varying a/L, where a is the radius of the growth tube and L is the transport length. Growth velocity data showed different trends at low and high aspect ratio, a result that does not support the velocity-aspect ratio trend predicted by theories. The system cannot be scaled on the basis of measurements done at a low aspect ratio. Some change in fluid flow behavior occurs in the growth tube as the aspect ratio increases.

  13. Growth of single crystals of mercuric iodide on the ground and in space

    SciTech Connect

    van den Berg, L.

    1993-06-01

    A short review is given of the methods by which mercuric iodide is prepared and purified to obtain material suitable for the growth of single crystals. Method used in our laboratory to grow single crystals up to 1,000 grams in weight from the vapor is discussed. Effects of gravity on the growth process are described. A crystal growth system suitable for operation in the reduced gravity environment of space has been designed and crystal growth experiments have been performed during the flights of Spacelab 3 (April 1985) and the International Microgravity Laboratory (January 1992). Structural quality and electronic properties of ground-based and space-grown crystals are compared.

  14. Acute kidney injury and disseminated intravascular coagulation due to mercuric chloride poisoning

    PubMed Central

    Dhanapriya, J.; Gopalakrishnan, N.; Arun, V.; Dineshkumar, T.; Sakthirajan, R.; Balasubramaniyan, T.; Haris, M.

    2016-01-01

    Mercury is a toxic heavy metal and occurs in organic and inorganic forms. Inorganic mercury includes elemental mercury and mercury salts. Mercury salts are usually white powder or crystals, and widely used in indigenous medicines and folk remedies in Asia. Inorganic mercury poisoning causes acute kidney injury (AKI) and gastrointestinal manifestations and can be life-threatening. We describe a case with unknown substance poisoning who developed AKI and disseminated intravascular coagulation (DIC). Renal biopsy showed acute tubular necrosis. Later, the consumed substance was proven to be mercuric chloride. His renal failure improved over time, and his creatinine normalized after 2 months. PMID:27194836

  15. In vitro toxicity of mercuric chloride on rabbit spermatozoa motility and cell membrane integrity.

    PubMed

    Slivkova, Jana; Massanyi, Peter; Pizzi, Flavia; Trandzik, Jozef; Roychoudhury, Shubhadeep; Lukac, Norbert; Dankova, Marianna; Almasiova, Viera

    2010-01-01

    In this in vitro study the effects of mercuric chloride on the motility and structural integrity of rabbit spermatozoa were investigated. The spermatozoa motility was evaluated using CASA method and Annexin analysis was used for detection of structural changes. The concentration of mercury in the medium varied from 5.0 to 83.3 microg HgCl(2)/mL. At Time 0 the highest motility was detected in the control group (67.09 +/- 8.72%). Motility in groups with mercury administration was lower in comparison with control. Significant differences were detected in groups with 50.0-83.3 microg HgCl(2)/mL (P < 0.001) at Time 0. After 60 and 120 minutes of incubation with mercuric chloride the motility significantly decreased almost in all experimental groups. Progressive motility had a decreasing trend in all experimental groups. At time 60 and 120 significant differences were noted in the group receiving 6.25-83.3 microg HgCl(2)/mL. Significant differences were detected in all experimental groups, except the group with the lowest mercuric chloride administration. The concentration-dependent decrease of spermatozoa progressive motility up to 50% of control was detected for groups receiving 50.0 - 83.3 microg HgCl(2)/mL at Time 0, for groups receiving 12.5-83.3 microg HgCl(2)/mL at Time 60 and 120, decreasing from 36.46 +/- 18.73% to 1.03 +/- 2.50%. Detailed evaluation of spermatozoa distance (DAP, DCL, and DSL) and velocity (VAP, VCL, and VSL) parameters as well as straightness (STR), linearity (LIN), wobble (WOB), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) of spermatozoa revealed decrease in groups with the highest mercury concentration in comparison with the control group at all time periods. Detection of spermatozoa with disordered membrane was carried out for groups with higher mercury concentrations and control, using Annexin analysis. Analysis showed higher occurrence of positive spermatozoa in the mercury exposed groups. Some Annexin

  16. Introduction to fifth international workshop on mercuric iodide nuclear radiation detectors

    SciTech Connect

    Schieber, M.

    1982-01-01

    Mercuric iodide is a wide bandgap semiconductor, with Eg approx. = 2.14 eV at room temperature. Therefore, HgI/sub 2/ is totally different from the well-studied, narrower gap, elemental semiconductors such as Si and Ge, and also different in its physical and chemical properties from the known semiconductor binary zinc-blend compounds such as GaAs or InP. The purpose of studies in the last decade was to further our understanding of HgI/sub 2/; recent progress is reported. (WHK)

  17. Experimental autologous immune deposit nephritis in rats associated with mercuric chloride administration.

    PubMed

    Kelchner, J; McIntosh, J R; Boedecker, E; Guggenheim, S; McIntosh, R M

    1976-09-15

    Serial administration of mercuric chloride to rats was followed by development of antibodies to tubular basement membrane and renal tubular epithelial antigen (RTE) and glomerulonephritis characterized by granular deposits of hosts IgG, C3 and RTE along the glomerular capillary walls. The glomerular fixed antibody was directed against RTE. These studies suggest that tubular injury by mercury may lead to release of RTE and autosensitization and subsequent antibody production to this antigen result in formation of and glomerular deposition of circulating immunopathogenic complexes (RTE-anti-RTE) and glomerular morphologic alterations.

  18. Growth of single crystals of mercuric iodide on the ground and in space

    NASA Astrophysics Data System (ADS)

    Vandenberg, L.

    1993-04-01

    A short review is given of the methods by which mercuric iodide is prepared and purified to obtain material suitable for the growth of single crystals. The method used in our laboratory to grow single crystals up to 1,000 grams in weight from the vapor is discussed. Effects of gravity on the growth process are described. A crystal growth system suitable for operation in the reduced gravity environment of space was designed and crystal growth experiments were performed during the flights of Spacelab 3 (April 1985) and the International Microgravity Laboratory (January 1992). Structural quality and electronic properties of ground-based and space-grown crystals are compared.

  19. A Highly Robust, Recyclable Displacement Assay for Mercuric Ions in Aqueous Solutions and Living Cells

    PubMed Central

    Liu, Dingbin; Wang, Shouju; Swierczewska, Magdalena; Huang, Xinglu; Bhirde, Ashwinkumar A.; Sun, Jiashu; Wang, Zhuo

    2012-01-01

    We designed a recyclable Hg2+ probe based on Rhodamine B isothiocyanate (RBITC) - poly (ethylene glycol) (PEG)-co-modified gold nanoparticles (AuNPs) with excellent robustness, selectivity and sensitivity. Based on a rational design, only Hg2+ can displace RBITC from the AuNP surfaces, resulting in a remarkable enhancement of RBITC fluorescence initially quenched by AuNPs. To maintain stability and monodispersity of AuNPs in real samples, thiol-terminated PEG was employed to bind with the remaining active sites of AuNPs. Besides, this displacement assay can be regenerated by resupplying free RBITC into the AuNPs solutions that were already used for detecting Hg2+. Importantly, the detection limit of this assay for Hg2+ (2.3 nM) was lower than the maximum limits guided by the United States Environmental Protection Agency as well as that permitted by the World Health Organization. The efficiency of this probe was demonstrated in monitoring Hg2+ in complex samples such as river water and living cells. PMID:23121626

  20. Purification, properties, and sequence of glycerol trinitrate reductase from Agrobacterium radiobacter.

    PubMed Central

    Snape, J R; Walkley, N A; Morby, A P; Nicklin, S; White, G F

    1997-01-01

    Glycerol trinitrate (GTN) reductase, which enables Agrobacterium radiobacter to utilize GTN and related explosives as sources of nitrogen for growth, was purified and characterized, and its gene was cloned and sequenced. The enzyme was a 39-kDa monomeric protein which catalyzed the NADH-dependent reductive scission of GTN (Km = 23 microM) to glycerol dinitrates (mainly the 1,3-isomer) with a pH optimum of 6.5, a temperature optimum of 35 degrees C, and no dependence on metal ions for activity. It was also active on pentaerythritol tetranitrate (PETN), on isosorbide dinitrate, and, very weakly, on ethyleneglycol dinitrate, but it was inactive on isopropyl nitrate, hexahydro-1,3,5-trinitro-1,3,5-triazine, 2,4,6-trinitrotoluene, ammonium ions, nitrate, or nitrite. The amino acid sequence deduced from the DNA sequence was homologous (42 to 51% identity and 61 to 69% similarity) to those of PETN reductase from Enterobacter cloacae, N-ethylmaleimide reductase from Escherichia coli, morphinone reductase from Pseudomonas putida, and old yellow enzyme from Saccharomyces cerevisiae, placing the GTN reductase in the alpha/beta barrel flavoprotein group of proteins. GTN reductase and PETN reductase were very similar in many respects except in their distinct preferences for NADH and NADPH cofactors, respectively. PMID:9401040

  1. Excitation and dissociation mechanisms in molecules with application to mercuric halide laser system

    SciTech Connect

    Spence, D.; Wang, R.G.; Dillon, M.A.

    1982-01-01

    Although the mercuric halide laser systems have received intensive study in recent years, being one of only two efficient electronic-transition lasers known, the precise collisional mechanisms leding to HgBr(B), formation and subsequent fluorescence are still imperfectly understood. The initial suggestion that direct collisional excitation of, say, HgBr/sub 2/, by electrons (analogous to photoionization), i.e., HgBr/sub 2/ + e ..-->.. HgBr(b) + Br + e, was the dominant mechanism, was temporarily abandoned when a measurement by Allison and Zare yielded a cross section of only < 1 x 10/sup -20/ cm/sup 2/ for low incident electron energy HgBr(B-x) fluorescence, much too small to explain the observed laser efficiency. Subsequent explanations for HgBr(B) formation included energy transfer from excited N/sub 2/ or rare gases, electronic recombination of HgBr/sub 2//sup +/, or dissociative electron attachment. Though it has recently been demonstrated that electronic energy transfer does play a role in HgBr(B) formation in the presence of N/sub 2/ or X/sub e/ buffers, modeling studies of e-beam sustained discharges have now conclusively shown that direct electron-impact excitation of mercuric halides, is indeed the dominant laser mechanism. The technique of electron-energy-loss spectroscopy was used to obtain pseudo-optical absorption spectra in HgBr/sub 2/ and HgCl/sub 2/. Results are presented and discussed. (WHK)

  2. A study of the homogeneity and deviations from stoichiometry in mercuric iodide

    SciTech Connect

    Burger, A.; Morgan, S.; He, C.; Silberman, E.; van den Berg, L.; Ortale, C.; Franks, L.; Schieber, M.

    1989-01-01

    We have been able to determine the deviations from stoichiometry of mercuric iodide (HgI/sub 2/) by using Differential Scanning Calorimetry (DSC). Mercury excess or iodine deficiency in mercuric iodide can be evaluated from the eutectic melting of HgI/sub 2/-- Hg/sub 2/I/sub 2/ at 235/degree/C which appears as an additional peak in DSC thermograms. I/sub 2/ excess can be found from the existence of the I/sub 2/--HgI/sub 2/ eutectic melting at 103/degree/C. An additional DSC peak appears in some samples around 112/degree/C that could be explained by the presence of iodine inclusions. Using Resonance Fluorescence Spectroscopy (RFS) we have been able to determine the presence of free I/sub 2/ that is released by samples during the heating at 120/degree/C (crystal growth temperature) thus giving additional support to the above DSC results. 19 refs., 6 figs., 2 tabs.

  3. Transdermal kinetics of a mercurous chloride beauty cream: an in vitro human skin analysis.

    PubMed

    Palmer, R B; Godwin, D A; McKinney, P E

    2000-01-01

    Crema de Belleza-Manning is a popular mercurous chloride-containing beauty cream used to smooth and lighten the complexion and treat acne. Hundreds of people in the Southwestern US border states have been identified with elevated (>20 microg/L) urine mercury levels believed to be secondary to using this cream. The kinetic characteristics of percutaneous mercury absorption are incompletely defined. The objective of this study was to determine the transdermal kinetics of two formulations of mercurous chloride from a beauty cream in an in vitro human skin model. A proprietary formulation and an aqueous formulation of the beauty cream were studied using modified Franz diffusion cells. Mercury content in the skin samples and the underlying diffusion buffer was determined using atomic absorption spectrophotometry. A rapid initial increase in mercury content both in the skin and the buffer was noted for both formulations. Mercury concentrations in the aqueous samples were significantly (p < 0.05) higher in both the skin and the diffusion buffer compared to parallel samples containing glycerol. Mercury was readily absorbed through the skin in this in vitro human skin model. The aqueous preparation had a markedly increased rate and extent of mercury absorption relative to the proprietary formulation.

  4. Antioxidant effect of Arabic gum against mercuric chloride-induced nephrotoxicity

    PubMed Central

    Gado, Ali M; Aldahmash, Badr A

    2013-01-01

    The effects of Arabic gum (AG) against nephrotoxicity of mercury (Hg), an oxidative-stress inducing substance, in rats were investigated. A single dose of mercuric chloride (5 mg/kg intraperitoneal injection) induced renal toxicity, manifested biochemically by a significant increase in serum creatinine, blood urea nitrogen, thiobarbituric acid reactive substances, and total nitrate/nitrite production in kidney tissues. In addition, reduced glutathione, glutathione peroxidase, and catalase enzymes in renal tissues were significantly decreased. Pretreatment of rats with AG (7.5 g/kg/day per oral administration), starting 5 days before mercuric chloride injection and continuing through the experimental period, resulted in a complete reversal of Hg-induced increase in creatinine, blood urea nitrogen, thiobarbituric acid reactive substances, and total nitrate/nitrite to control values. Histopathologic examination of kidney tissues confirmed the biochemical data; pretreatment of AG prevented Hg-induced degenerative changes of kidney tissues. These results indicate that AG is an efficient cytoprotective agent against Hg-induced nephrotoxicity by a mechanism related at least in part to its ability to decrease oxidative and nitrosative stress and preserve the activity of antioxidant enzymes in kidney tissues. PMID:24174869

  5. Antioxidant effect of Arabic gum against mercuric chloride-induced nephrotoxicity.

    PubMed

    Gado, Ali M; Aldahmash, Badr A

    2013-01-01

    The effects of Arabic gum (AG) against nephrotoxicity of mercury (Hg), an oxidative-stress inducing substance, in rats were investigated. A single dose of mercuric chloride (5 mg/kg intraperitoneal injection) induced renal toxicity, manifested biochemically by a significant increase in serum creatinine, blood urea nitrogen, thiobarbituric acid reactive substances, and total nitrate/nitrite production in kidney tissues. In addition, reduced glutathione, glutathione peroxidase, and catalase enzymes in renal tissues were significantly decreased. Pretreatment of rats with AG (7.5 g/kg/day per oral administration), starting 5 days before mercuric chloride injection and continuing through the experimental period, resulted in a complete reversal of Hg-induced increase in creatinine, blood urea nitrogen, thiobarbituric acid reactive substances, and total nitrate/nitrite to control values. Histopathologic examination of kidney tissues confirmed the biochemical data; pretreatment of AG prevented Hg-induced degenerative changes of kidney tissues. These results indicate that AG is an efficient cytoprotective agent against Hg-induced nephrotoxicity by a mechanism related at least in part to its ability to decrease oxidative and nitrosative stress and preserve the activity of antioxidant enzymes in kidney tissues.

  6. Zeatin reductase in Phaseolus embryos

    SciTech Connect

    Martin, R.C.; Mok, David, W.S.; Mok, M.C. )

    1989-04-01

    Zeatin was converted to O-xylosylzeatin in embryos of Phaseolus vulgaris . O-xylosyldihydrozeatin was also identified as a zeatin metabolite. Incubation of embryo extracts with {sup 14}C-zeatin and {sup 14}C-O-xylosylzeatin revealed that reduction preceeds the O-xylosylation of zeatin. An enzyme responsible for reducing the N{sup 6}-side chain was isolated and partially purified using ammonium sulfate fractionation and affinity, gel filtration and anion exchange chromatography. The NADPH dependent reductase was zeatin specific and did not recognize cis-zeatin, ribosylzeatin, i{sup 6}Ade or i{sup 6}Ado. Two forms of the reductase could be separated by either gel filtration or anion exchange HPLC. The HMW isozyme (Mr. 55,000) eluted from the anion exchange column later than the LMW isozyme (Mr. 25,000). Interspecific differences in zeatin reductase activity were also detected.

  7. Isolated menthone reductase and nucleic acid molecules encoding same

    DOEpatents

    Croteau, Rodney B; Davis, Edward M; Ringer, Kerry L

    2013-04-23

    The present invention provides isolated menthone reductase proteins, isolated nucleic acid molecules encoding menthone reductase proteins, methods for expressing and isolating menthone reductase proteins, and transgenic plants expressing elevated levels of menthone reductase protein.

  8. Ascorbate free radical reductases and diaphorases in soluble fractions of the human lens.

    PubMed

    Bando, M; Obazawa, H

    1995-12-01

    Major and minor ascorbate free radical (AFR) reductases, with diaphorase activity, and three other diaphorases were separated from the human lens soluble fraction by DEAE-cellulose ion-exchange column chromatography. They were characterized for adsorptivity to ion-exchange and 5'AMP-Sepharose 4B affinity columns, kinetic properties, and substrate specificity. The latter diaphorases were closely correlated with NADH-cytochrome beta 5 reductase. The major and minor AFR reductases were regarded as a major diaphorase group different from two ubiquitous diaphorases, i.e., NADH-cytochrome beta 5 reductase and DT-diaphorase. A major AFR reductase was partially purified approximately 50 fold over the lens soluble fraction by ion-exchange, affinity, and gel filtration (Sephacryl S-200 HR) column chromatography. From the partially purified enzyme, 2 bands, one sharp and one diffuse, were obtained by native polyacrylamide gel electrophoresis. Two proteins, of 20 and 24 kDa, were identified in the active enzyme bands by SDS-polyacrylamide gel electrophoresis. This suggests that the 20 and/or 24 kDa proteins may be components of the major AFR reductase.

  9. Histochemical localization of nitrate reductase.

    PubMed

    Vaughn, K C; Duke, S O

    1981-01-01

    NADH-dependent nitrate reductase (E.C. 1.6.6.1) was ultrastructurally localized in norflurazon-treated and control soybean cotyledons [Glycine max (L.) Merr.] by a method based upon the increase in osmiophilia due to the formation of an azo dye. The reaction product was observed in small vesicles throughout the cytoplasm. An apparent transport of nitrite to the plastid, the site of nitrite reduction, may occur through fusion of the nitrite-containing vesicles with the chloroplast envelope. Plants grown in tungstate lacked nitrate reductase activity as measured by standard assay procedures, and showed no increase in osmiophilia, suggesting a degree of specificity of this cytochemical procedure.

  10. ROLE OF SURFACE FUNCTIONAL GROUPS IN THE CAPTURE OF ELEMENTAL MERCURY AND MERCURIC CHLORIDE BY ACTIVATED CARBONS

    EPA Science Inventory

    The paper discusses using a laboratory-scale, fixed bed apparatus to study the role of surface functional groups (SFGs) in the capture of mercuric chloride (HgC12) and elemental mercury (Hgo) in nitrogen (N2) prior to flue gas atmosphere studies. The study focused on two activat...

  11. Growth of high quality mercurous halide single crystals by physical vapor transport method for AOM and radiation detection applications

    NASA Astrophysics Data System (ADS)

    Amarasinghe, Priyanthi M.; Kim, Joo-Soo; Chen, Henry; Trivedi, Sudhir; Qadri, Syed B.; Soos, Jolanta; Diestler, Mark; Zhang, Dajie; Gupta, Neelam; Jensen, Janet L.; Jensen, James

    2016-09-01

    Single crystals of mercurous halide were grown by physical vapor transport method (PVT). The orientation and the crystalline quality of the grown crystals were determined using high resolution x-ray diffraction (HRXRD) technique. The full width at half maximum (FWHM) of the grown mercurous bromide crystals was measured to be 0.13 degrees for (004) reflection, which is the best that has been achieved so far for PVT grown mercurous halide single crystals. The extended defects of the crystals were also analyzed using high resolution x-ray diffraction topography. Preliminary studies were carried out to evaluate the performance of the crystals on acousto-optic modulator (AOM) and gamma-ray detector applications. The results indicate the grown mercurous halide crystals are excellent materials for acousto-optic modulator device fabrication. The diffraction efficiencies of the fabricated AOM device with 1152 and 1523 nm wavelength lasers polarizing parallel to the acoustic wave were found to be 35% and 28%, respectively. The results also indicate the grown crystals are a promising material for gamma-ray detector application with a very high energy resolution of 1.86% FWHM.

  12. ROLE OF SURFACE FUNCTIONAL GROUPS IN THE CAPTURE OF ELEMENTAL MERCURY AND MERCURIC CHLORIDE BY ACTIVATED CARBONS

    EPA Science Inventory

    The paper discusses using a laboratory-scale, fixed bed apparatus to study the role of surface functional groups (SFGs) in the capture of mercuric chloride (HgC12) and elemental mercury (Hgo) in nitrogen (N2) prior to flue gas atmosphere studies. The study focused on two activat...

  13. Use of mercuric iodide X-ray detectors with alpha backscattering spectrometers for space applications

    NASA Technical Reports Server (NTRS)

    Iwanczyk, J. S.; Wang, Y. J.; Dorri, N.; Dabrowski, A. J.; Economou, T. E.

    1991-01-01

    The authors present X-ray fluorescence (XRF) spectra of different extraterrestrial samples taken with a mercuric iodide (HgI2) spectrometer inserted into an alpha backscattering instrument identical to that used in the Soviet Phobos mission. The results obtained with the HgI2 ambient temperature detector are compared with those obtained using an Si(Li) cryogenically cooled detector. Efforts to design an optimized instrument for space application are also described. The results presented indicate that the energy resolution and sensitivity of HgI2 detectors are adequate to meet the performance needs of a number of proposed space applications, particularly those in which cooled silicon X-ray detectors are impractical or even not usable, such as for the target science programs on geoscience opportunities for lunar surface, Mars surface, and other comet and planetary missions being planned by NASA and ESA.

  14. Inhibition of implantation caused by methylmercury and mercuric chloride in mouse embryos in vivo

    SciTech Connect

    Kajiwara, Yuji; Inouye, Minoru

    1992-10-01

    Methylmercury, an environmental pollutant, produces a wide spectrum of fetotoxic effects in men and laboratory animals. Experimental studies have shown that the exposure to methylmercury in the gestation period causes fetal death, gross malformation, growth retardation of the fetuses, and stillbirth. Although the effects of methylmercury on fetuses have been well documented, only a few experiments have been performed on the embryo toxicity at the early gestation periods. Because the embryos at preimplantation period are known to be highly sensitive to methylmercury in vitro and in vivo, in the present experiment, the embryonic development after implantation was investigated following treatment with methylmercury during the preimplantation period. Since the previous report showed that methylmercury and inorganic mercury were different in their manifestation of toxicity on preimplantation and mercuric chloride on embryos were investigated in vivo in the present study. 22 refs., 3 figs., 3 tabs.

  15. A study of mercuric oxide and zinc-air battery life in hearing aids.

    PubMed

    Sparkes, C; Lacey, N K

    1997-09-01

    The requirement to phase out mercuric oxide (mercury) batteries on environmental grounds has led to the widespread introduction of zinc-air technology. The possibility arises that high drain hearing aids may not be adequately catered for by zinc-air cells, leading to poor performance. This study investigated the hearing aid user's ability to perceive differences between zinc-air and mercury cells in normal everyday usage. The data was collected for 100 experienced hearing aid users in field trials. Users report 50 per cent greater life for zinc-air cells in high power aids and 28 per cent in low power aids. The average life of the zinc-air cells range from 15 days in high power to 34 days in low power aids. Users are able to perceive a difference in sound quality in favour of zinc-air cells for low and medium power aids. The hearing aid population is not disadvantaged by phasing out mercury cells.

  16. Detection of mercuric chloride by photofragment emission using a frequency-converted fiber amplifier

    NASA Astrophysics Data System (ADS)

    Hoops, Alexandra A.; Reichardt, Thomas A.; Kliner, Dahv. A. V.; Koplow, Jeffrey P.; Moore, Sean W.

    2007-07-01

    A real-time, noninvasive approach for detecting trace amounts of vapor-phase mercuric chloride (HgCl2) in combustion flue gas is demonstrated using a near-infrared pulsed fiber amplifier that is frequency converted to the ultraviolet. Excitation of the HgCl2(1∏1u ← 1∑1g+) transition at 213 nm generates 253.7 nm emission from the Hg (63P1) photoproduct that is proportional to the concentration of HgCl2. A measured quadratic dependence of the HgCl2 photofragment emission (PFE) signal on the laser irradiance indicates that the photodissociation process involves two-photon excitation. Additionally, low concentrations of HgCl2 are detected with the PFE approach in an environment characteristic of coal-fired power-plant flue gas using this compact solid-state laser source. A detection limit of 0.7 ppb is extrapolated from these results.

  17. Reproductivity of Japanese quail fed mercuric chloride in the absence of vitamin D

    USGS Publications Warehouse

    Hill, E.F.; Soares, J.H.

    1977-01-01

    Mercuric chloride (HgCl2) was tested at 16 p.p.m. Hg for vitamin D sparing activity by presenting it dietarily in the presence and absence of 25-hydroxycholecalciferol (25-HCC) to Japanese quail (Coturnix c. japonica) for 25 days. No gross signs characteristic of mercury poisoning were observed, but some predictable effects of vitamin D deficiency on avian reproduction were manifested within 10 days. Rate of lay, egg shell thickness, and hatchability of fertile eggs decreased markedly for birds on vitamin D-deficient diets. Shell-less eggs were laid by these birds after 20 days and laying stopped entirely on the 23rd day. Laying resumed within 5 days after diets were refortified with 25-HCC. There was no detectable interaction between HgCl2 and vitamin D.

  18. Improvement in pixel signal uniformity of polycrystalline mercuric iodide films for digital X-ray imaging

    NASA Astrophysics Data System (ADS)

    Oh, Kyungmin; Kim, Jinseon; Shin, Jungwook; Heo, Seunguk; Cho, Gyuseok; Kim, Daekuk; Park, Jigoon; Nam, Sanghee

    2014-03-01

    We investigated polycrystalline mercuric iodide (HgI2) that exhibits uniform pixel signals for its use in digital X-ray imaging. To fabricate thin polycrystalline HgI2 films, the particle-in-binder (PIB) method is used because it enables the fabrication of X-ray conversion films at a low temperature and a normal pressure. Moreover, it has a large-scale deposition capacity at a low cost. Although the thin layers fabricated by the PIB method have such advantages, they are chemically unstable and show poor reproducibility and nonuniform X-ray response. To solve these problems, in this study, additional physical and chemical treatments were performed along with the PIB method after taking the size confinement effect of photoconductive particles into consideration. Morphological and electrical properties were measured to investigate the effects of the physical and chemical treatments.

  19. A review of recent measurements of optical and thermal properties of. alpha. -mercuric iodide

    SciTech Connect

    Burger, A.; Morgan, S.H.; Silberman, E. . Dept. of Physics); Nason, D.; Cheng, A.Y. . Santa Barbara Operations)

    1991-01-01

    The knowledge of the physical properties of a crystal and their relation to the nature and content of defects are essential for both applications and fundamental reasons. Alpha-mercuric iodide ({alpha}-HgI{sub 2}) is a material which was found important applications as room temperature X-ray and gamma ray detectors. Some recent thermal and optical measurements of this material, using the samples of improved crystallinity which are now available, are reviewed below. Heretofore, these properties have received less attention than the mechanical and electrical properties, particularly at elevated temperatures. In the technology of {alpha}-HgI{sub 2} where there is a continuing motivation to obtain larger single crystals without compromising the material quality, a better knowledge of the thermal and optical properties may lead to improvements in the processes of material purification, crystal growth and device fabrication.

  20. Photoluminescence variations associated with the deposition of palladium electrical contacts on detector-grade mercuric iodide

    SciTech Connect

    Wong, D.; Bao, X.J.; Schlesinger, T.E.; James, R.B.; Cheng, A.; Ortale, C.; van den Berg, L.

    1988-10-17

    Specimens of mercuric iodide with evaporated semitransparent palladium contacts have been studied using low-temperature photoluminescence spectroscopy. Distinct differences were found between spectra taken from beneath the Pd contacts and those taken from regions on the HgI/sub 2/ sample that were masked during the Pd deposition, indicating that contact fabrication can change the defect structure near the contact/substrate interface. Comparison of the spectra from spots beneath the contacts with spectra from bulk material specimens and HgI/sub 2/ detectors graded in terms of their nuclear detection performance suggests that the processing steps used to deposit electrical contacts and the choice of contact material may have a significant influence on detector performance.

  1. Efficacy and safety of mercuric oxide in the treatment of bacterial blepharitis.

    PubMed Central

    Hyndiuk, R A; Burd, E M; Hartz, A

    1990-01-01

    A double-masked, placebo-controlled, randomized study was done to assess the safety and clinical and quantitative microbiologic efficacy of 1% mercuric oxide (yellow) ophthalmic ointment in the treatment of eyelid infections, i.e., bacterial blepharitis. A total of 39 patients with bacterial counts and clinical signs indicative of eyelid infection were treated twice daily for 7 days. Clinical biomicroscopic examination and quantitative microbiologic cultures were done just prior to initiation of treatment and again on days 3 and 7. Statistical analysis revealed a significant improvement in the clinical signs, bacterial count, cure rate, and improvement rate for subjects taking the active medication, compared with those taking the placebo on days 3 and 7. In addition, the medication was well tolerated. PMID:2344168

  2. Physical Vapor Transport of Mercurous Chloride Crystals: Design of a Microgravity Experiment

    NASA Technical Reports Server (NTRS)

    Duval, W, M. B.; Singh, N. B.; Glicksman, M. E.

    1997-01-01

    Flow field characteristics predicted from a computational model show that the dynamical state of the flow, for practical crystal growth conditions of mercurous chloride, can range from steady to unsteady. Evidence that the flow field can be strongly dominated by convection for ground-based conditions is provided by the prediction of asymmetric velocity profiles bv the model which show reasonable agreement with laser Doppler velocimetry experiments in both magnitude and planform. Unsteady flow is shown to be correlated with a degradation of crystal quality as quantified by light scattering pattern measurements, A microgravity experiment is designed to show that an experiment performed with parameters which yield an unsteady flow becomes steady (diffusive-advective) in a microgravity environment of 10(exp -3) g(sub 0) as predicted by the model, and hence yields crystals with optimal quality.

  3. Detection of mercuric chloride by photofragment emission using a frequency-converted fiber amplifier.

    PubMed

    Hoops, Alexandra A; Reichardt, Thomas A; Kliner, Dahv A V; Koplow, Jeffrey P; Moore, Sean W

    2007-07-01

    A real-time, noninvasive approach for detecting trace amounts of vapor-phase mercuric chloride (HgCl(2)) in combustion flue gas is demonstrated using a near-infrared pulsed fiber amplifier that is frequency converted to the ultraviolet. Excitation of the HgCl(2) ([see text]) transition at 213 nm generates 253.7 nm emission from the Hg (6(3)P(1)) photoproduct that is proportional to the concentration of HgCl(2). A measured quadratic dependence of the HgCl(2) photofragment emission (PFE) signal on the laser irradiance indicates that the photodissociation process involves two-photon excitation. Additionally, low concentrations of HgCl(2) are detected with the PFE approach in an environment characteristic of coal-fired power-plant flue gas using this compact solid-state laser source. A detection limit of 0.7 ppb is extrapolated from these results.

  4. Specific labeling and partial inactivation of cytochrome oxidase by fluorescein mercuric acetate.

    PubMed

    Stonehuerner, J; O'Brien, P; Kendrick, L; Hall, J; Millett, F

    1985-09-25

    Addition of 1 eq of fluorescein mercuric acetate (FMA) to beef heart cytochrome oxidase was found to inhibit the steady-state electron transfer activity by 50%, but further additions up to 10 eq had no additional effect on activity. The partial inhibition caused by FMA is thus similar to that observed with other mercury compounds (Mann, A. J., and Auer, H. E. (1980) J. Biol. Chem. 255, 454-458). The fluorescence of FMA was quenched by a factor of 10 upon binding to cytochrome oxidase, consistent with the involvement of a sulfhydryl group. However, addition of mercuric chloride to FMA-cytochrome oxidase resulted in an increase in fluorescence, suggesting that FMA was displaced from the high affinity binding site. Cytochrome c binding to FMA-cytochrome oxidase resulted in a 10% decrease in the fluorescence, possibly caused by Forster energy transfer from FMA to the cytochrome c heme. The binding site for FMA in cytochrome oxidase was investigated by carrying out sodium dodecyl sulfate gel electrophoresis under progressively milder dissociation conditions. When FMA-cytochrome oxidase was dissociated with 3% sodium dodecyl sulfate and 6 M urea, FMA was predominantly bound to subunit II following electrophoresis. However, when the dissociation was carried out at 4 degrees C in the absence of urea with progressively smaller amounts of lithium dodecyl sulfate, the labeling of subunit II decreased and that of subunit I increased. These experiments demonstrate that mercury compounds bind to a high affinity site on cytochrome oxidase, possibly located in subunit I, but then migrate to subunit II under the normal sodium dodecyl sulfate gel electrophoresis conditions. A definitive assignment of the high affinity binding site in the native enzyme cannot be made, however, because it is possible that mercury compounds can migrate from one sulfhydryl to another under even the mildest electrophoresis conditions.

  5. State of the art and potential applications of Mercuric Iodide radiation detectors

    SciTech Connect

    Holzer, A.

    1982-01-01

    Mercuric Iodide (HgI/sub 2/) has been recognized as the best room temperature solid-state, X-ray detection material presently available. While the detection performance of Mercuric Iodide is not as good as liquid nitrogen cooled Si(Li) or Germanium it is already good enough to meet the requirements of several special applications where the simplicity and convenience of room temperature operation are important. The high atomic numbers of Hg and I (80, 53) enable efficient absorption of radiation, and the wide band gap (2.13 eV versus 1.12 eV for silicon) allows operation at room temperature without any significant thermal noise. Poor hole collection, resulting from deep hole trapping centers, is the main limitation in the use of HgI/sub 2/ for high energy (>60 keV) gamma-ray detection, but fortunately this is not a problem for detecting the 5--20 keV X-radiation normally used in crystallography. These lower energy X-rays are absorbed within a few microns of the negative electrode and so the holes do not contribute significantly to the pulse. In such cases, very good energy resolution can be obtained. The present performance characteristics for detection of X-rays: i.e., good energy resolution, long-term stability, and the lack of polarization effects: open a wide range of applications for HgI/sub 2/ detectors. This paper will focus on the different methods used to grow HgI/sub 2/ crystals and on how the method of growth is reflected in detector performance. The state of the art of HgI/sub 2/ detector capabilities is discussed and several of the most attractive applications are pointed out.

  6. The aldo-keto reductases (AKRs): Overview.

    PubMed

    Penning, Trevor M

    2015-06-05

    The aldo-keto reductase (AKR) protein superfamily contains >190 members that fall into 16 families and are found in all phyla. These enzymes reduce carbonyl substrates such as: sugar aldehydes; keto-steroids, keto-prostaglandins, retinals, quinones, and lipid peroxidation by-products. Exceptions include the reduction of steroid double bonds catalyzed by AKR1D enzymes (5β-reductases); and the oxidation of proximate carcinogen trans-dihydrodiol polycyclic aromatic hydrocarbons; while the β-subunits of potassium gated ion channels (AKR6 family) control Kv channel opening. AKRs are usually 37kDa monomers, have an (α/β)8-barrel motif, display large loops at the back of the barrel which govern substrate specificity, and have a conserved cofactor binding domain. AKRs catalyze an ordered bi bi kinetic mechanism in which NAD(P)H cofactor binds first and leaves last. In enzymes that favor NADPH, the rate of release of NADP(+) is governed by a slow isomerization step which places an upper limit on kcat. AKRs retain a conserved catalytic tetrad consisting of Tyr55, Asp50, Lys84, and His117 (AKR1C9 numbering). There is conservation of the catalytic mechanism with short-chain dehydrogenases/reductases (SDRs) even though they show different protein folds. There are 15 human AKRs of these AKR1B1, AKR1C1-1C3, AKR1D1, and AKR1B10 have been implicated in diabetic complications, steroid hormone dependent malignancies, bile acid deficiency and defects in retinoic acid signaling, respectively. Inhibitor programs exist world-wide to target each of these enzymes to treat the aforementioned disorders. Inherited mutations in AKR1C and AKR1D1 enzymes are implicated in defects in the development of male genitalia and bile acid deficiency, respectively, and occur in evolutionarily conserved amino acids. The human AKRs have a large number of nsSNPs and splice variants, but in many instances functional genomics is lacking. AKRs and their variants are now poised to be interrogated using

  7. Nitrate reductase from Rhodopseudomonas sphaeroides.

    PubMed Central

    Kerber, N L; Cardenas, J

    1982-01-01

    The facultative phototroph Rhodopseudomonas sphaeroides DSM158 was incapable of either assimilating or dissimilating nitrate, although the organism could reduce it enzymatically to nitrite either anaerobically in the light or aerobically in the dark. Reduction of nitrate was mediated by a nitrate reductase bound to chromatophores that could be easily solubilized and functioned with chemically reduced viologens or photochemically reduced flavins as electron donors. The enzyme was solubilized, and some of its kinetic and molecular parameters were determined. It seemed to be nonadaptive, ammonia did not repress its synthesis, and its activity underwent a rapid decline when the cells entered the stationary growth phase. Studies with inhibitors and with metal antagonists indicated that molybdenum and possibly iron participate in the enzymatic reduction of nitrate. The conjectural significance of this nitrate reductase in phototrophic bacteria is discussed. PMID:6978883

  8. Pyrroline-5-Carboxylate Reductase in Chlorella autotrophica and Chlorella saccharophila in Relation to Osmoregulation.

    PubMed

    Laliberté, G; Hellebust, J A

    1989-11-01

    Pyrroline-5-carboxylate (P5C) reductase (EC 1.5.1.2), which catalyzes the reduction of P5C to proline, was partially purified from two Chlorella species; Chlorella autotrophica, a euryhaline marine alga that responds to increases in salinity by accumulating proline and ions, and Chlorella saccharophila, which does not accumulate proline for osmoregulation. From the elution profile of this enzyme from an anion exchange column in Tris-HCl buffer (pH 7.6), containing sorbitol and glycine betaine, it was shown that P5C reductase from C. autotrophica was a neutral protein whereas the enzyme from C. saccharophila was negatively charged. The kinetic mechanisms of the reductase was characteristic of a ping-pong mechanism with double competitive substrate inhibition. Both enzymes showed high specificity for NADH as cofactor. The affinities of the reductases for their substrates did not change when the cells were grown at different salinities. In both algae, the apparent K(m) values of the reductase for P5C and NADH were 0.17 and 0.10 millimolar, respectively. A fourfold increase in maximal velocity of the reductase was observed when C. autotrophica was transferred from 50 to 150% artificial sea water. Even though the reductase was inhibited by NaCl, KCl, and proline, it still showed appreciable activity in the presence of these compounds at molar concentrations. A possible role for the regulation of proline synthesis at the step catalyzed by P5C reductase is discussed in relation to the specificity of P5C reductase for NADH and its responses to salt treatments.

  9. Pyrroline-5-Carboxylate Reductase in Chlorella autotrophica and Chlorella saccharophila in Relation to Osmoregulation 1

    PubMed Central

    Laliberté, Gilles; Hellebust, Johan A.

    1989-01-01

    Pyrroline-5-carboxylate (P5C) reductase (EC 1.5.1.2), which catalyzes the reduction of P5C to proline, was partially purified from two Chlorella species; Chlorella autotrophica, a euryhaline marine alga that responds to increases in salinity by accumulating proline and ions, and Chlorella saccharophila, which does not accumulate proline for osmoregulation. From the elution profile of this enzyme from an anion exchange column in Tris-HCl buffer (pH 7.6), containing sorbitol and glycine betaine, it was shown that P5C reductase from C. autotrophica was a neutral protein whereas the enzyme from C. saccharophila was negatively charged. The kinetic mechanisms of the reductase was characteristic of a ping-pong mechanism with double competitive substrate inhibition. Both enzymes showed high specificity for NADH as cofactor. The affinities of the reductases for their substrates did not change when the cells were grown at different salinities. In both algae, the apparent Km values of the reductase for P5C and NADH were 0.17 and 0.10 millimolar, respectively. A fourfold increase in maximal velocity of the reductase was observed when C. autotrophica was transferred from 50 to 150% artificial sea water. Even though the reductase was inhibited by NaCl, KCl, and proline, it still showed appreciable activity in the presence of these compounds at molar concentrations. A possible role for the regulation of proline synthesis at the step catalyzed by P5C reductase is discussed in relation to the specificity of P5C reductase for NADH and its responses to salt treatments. PMID:16667157

  10. Δ1-Pyrroline-5-carboxylate reductase from Arabidopsis thaliana: stimulation or inhibition by chloride ions and feedback regulation by proline depend on whether NADPH or NADH acts as co-substrate.

    PubMed

    Giberti, Samuele; Funck, Dietmar; Forlani, Giuseppe

    2014-05-01

    Δ(1)-pyrroline-5-carboxylate (P5C) reductase (P5CR) catalyses the final step of proline synthesis in plants. In Arabidopsis thaliana, protein levels are correlated neither to the corresponding mRNA copy numbers, nor to intracellular proline concentrations. The occurrence of post-translational regulatory mechanisms has therefore been hypothesized, but never assessed. The purification of A. thaliana P5CR was achieved through either a six-step protocol from cultured cells, or heterologous expression of AtP5CR in Escherichia coli. The protein was characterized with respect to structural, kinetic, and biochemical properties. P5CR was able to use either NADPH or NADH as the electron donor, with contrasting affinities and maximum reaction rates. The presence of equimolar concentrations of NADP(+) completely suppressed the NADH-dependent activity, whereas the NADPH-dependent reaction was mildly affected. Proline inhibited only the NADH-dependent reaction. At physiological values, increasing concentrations of salt progressively inhibited the NADH-dependent activity, but were stimulatory of the NADPH-dependent reaction. The biochemical properties of A. thaliana P5CR suggest a complex regulation of enzyme activity by the redox status of the pyridine nucleotide pools, and the concentrations of proline and chloride in the cytosol. Data support a to date underestimated role of P5CR in controlling stress-induced proline accumulation.

  11. Copper-dependent inhibition and oxidative inactivation with affinity cleavage of yeast glutathione reductase.

    PubMed

    Murakami, Keiko; Tsubouchi, Ryoko; Fukayama, Minoru; Yoshino, Masataka

    2014-06-01

    Effects of copper on the activity and oxidative inactivation of yeast glutathione reductase were analyzed. Glutathione reductase from yeast was inhibited by cupric ion and more potently by cuprous ion. Copper ion inhibited the enzyme noncompetitively with respect to the substrate GSSG and NADPH. The Ki values of the enzyme for Cu(2+) and Cu(+) ion were determined to be 1 and 0.35 μM, respectively. Copper-dependent inactivation of glutathione reductase was also analyzed. Hydrogen peroxide and copper/ascorbate also caused an inactivation with the cleavage of peptide bond of the enzyme. The inactivation/fragmentation of the enzyme was prevented by addition of catalase, suggesting that hydroxyl radical produced through the cuprous ion-dependent reduction of oxygen is responsible for the inactivation/fragmentation of the enzyme. SDS-PAGE and TOF-MS analysis confirmed eight fragments, which were further determined to result from the cleavage of the Met17-Ser18, Asn20-Thr21, Glu251-Gly252, Ser420-Pro421, Pro421-Thr422 bonds of the enzyme by amino-terminal sequencing analysis. Based on the kinetic analysis and no protective effect of the substrates, GSSG and NADPH on the copper-mediated inactivation/fragmentation of the enzyme, copper binds to the sites apart from the substrate-sites, causing the peptide cleavage by hydroxyl radical. Copper-dependent oxidative inactivation/fragmentation of glutathione reductase can explain the prooxidant properties of copper under the in vivo conditions.

  12. Alternating metastable/stable pattern in the mercuric iodide crystal formation outside the Ostwald Rule of Stages.

    PubMed

    Ayass, Mahmoud M; Abi Mansour, Andrew; Al-Ghoul, Mazen

    2014-09-11

    We report a reaction-diffusion system in which two initially separated electrolytes, mercuric chloride (outer) and potassium iodide (inner), interact in a solid hydrogel media to produce a propagating front of mercuric iodide precipitate. The precipitation process is accompanied by a polymorphic transformation of the kinetically favored (unstable) orange, (metastable) yellow, and (thermodynamically stable) red polymorphs of HgI2. The sequence of crystal transformation is confirmed to agree with the Ostwald Rule of Stages. However, a region is found of initial inner iodide concentration, where a stationary pattern of alternating metastable/stable crystals is formed. A theoretical model based on reaction diffusion coupled to a special nucleation and growth mechanism is proposed. Its numerical solution is shown to reproduce the experimental results.

  13. 1984 State of the art of the technology of mercuric iodide x-ray and gamma radiation detectors

    SciTech Connect

    Schieber, M.; Schnepple, W.

    1983-10-01

    The present state of the art of mercuric iodide technology is reviewed. Recent progress is reported in the use of HgI/sub 2/ in high energy resolution x-ray and gamma ray spectrometers which operate at room temperature. Purification of starting materials, methods of crystal growth, detector fabrication, and characterization methods used for HgI/sub 2/ are described, and some applications of HgI/sub 2/ detectors to various device systems are given.

  14. Mercuric iodide research and development in support of DOE Historically Black Colleges and University Program. Semiannual technical progress report

    SciTech Connect

    George, M.A.; Zheng, Y.; Salary, L.; Chen, K.T.; Burger, A.

    1994-10-31

    This report describes the progress achieved during the first six months of the program. The different subjects studied were: zone refining experiments of mercuric iodide to establish optimum refining parameters and produce purified material; development of surface reflection spectroscopy as a method to measure crystal surface temperatures, with emphasis on investigation the potential of using optical multichannel analysis; optical methods for measuring iodine vapor during physical vapor transport of HgI{sub 2}; and atomic force microscopy studies.

  15. Fatty acyl-CoA reductase

    SciTech Connect

    Reiser, Steven E.; Somerville, Chris R.

    1998-12-01

    The present invention relates to bacterial enzymes, in particular to an acyl-CoA reductase and a gene encoding an acyl-CoA reductase, the amino acid and nucleic acid sequences corresponding to the reductase polypeptide and gene, respectively, and to methods of obtaining such enzymes, amino acid sequences and nucleic acid sequences. The invention also relates to the use of such sequences to provide transgenic host cells capable of producing fatty alcohols and fatty aldehydes.

  16. Superdiffusive cusp-like waves in the mercuric iodide precipitate system and their transition to regular reaction bands.

    PubMed

    Ayass, Mahmoud M; Al-Ghoul, Mazen

    2014-06-05

    We report a two-dimensional (2D) reaction-diffusion system that exhibits a superdiffusive propagating wave with anomalous cusp-like contours. This wave results from a leading precipitation reaction (wavefront) and a trailing redissolution (waveback) between initially separated mercuric chloride and potassium iodide to produce mercuric iodide precipitate (HgI2) in a thin sheet of a solid hydrogel (agar) medium. The propagation dynamics is accompanied by continuous polymorphic transformations between the metastable yellow crystals and the stable red crystals of HgI2. We study the dynamics of wavefront and waveback propagation that reveals interesting anomalous superdiffusive behavior without the influence of external enhancement. We find that a transition from superdiffusive to subdiffusive dynamics occurs as a function of outer iodide concentration. Inner mercuric concentrations lead to the transition from the anomalous cusp-like to cusp-free regular bands. While gel concentration affects the speed of propagation of the wave, it has no effect on its shape or on its superdiffusive dynamics. Microscopically, we show that the macroscopic wave propagation and polymorphic transformations are accompanied by an Ostwald ripening mechanism in which larger red HgI2 crystals are formed at the expense of smaller yellow HgI2 crystals.

  17. Abondance et stratification verticale des elements dans l'atmosphere des etoiles mercure-manganese

    NASA Astrophysics Data System (ADS)

    Thiam, Mouhamadou

    Les étoiles mercure-manganèse appartiennent à la famille des étoiles chimiquement particulières. Elles présentent d'importants excès de mercure et de manganèse par rapport au soleil. Ces anomalies d'abondance sont généralement expliquées par la théorie de la diffusion atomique, jouant possiblementun rôle important au sein de leurs atmosphères. Cette thèse a pour but, dans un premier temps, de déterminer les abondances d'un grand nombre d'éléments (C, O, Mg, Si, P, S, Ca, Sc, Ti, Cr, Mn, Fe, Sr, Hg) peuplant l'atmosphère de quatre étoiles mercure-manganèse (HD 71066, HD 175640, HD 178065 et HD 221507). La seconde étape consiste à vérifier la présence d'une quelconque dépendance de l'abondance de ces éléments par rapport à la profondeur de formation des raies. Une variation de l'abondance par rapport à la profondeur, c'est-à-dire stratification, apporterait la preuve observationnelle de la présence de la diffusion atomique au sein de l'atmosphère de ces étoiles, ainsi que des contraintes servant à l'amélioration des modèles théoriques. Les spectres des étoiles étudiées sont obtenus à partir de l'archive de l'ESO- UVES. Les raies investiguées se situent à des longueurs d'onde plus grandes que celle du saut de Balmer oû le signal sur bruit, après normalisation, est supérieur à 300. Parmi les quatre étoiles investiguées, uniquement de la stratification du manganèse a été découverte dans l'atmosphère de l'étoile HD 178065. L'abondance du manganèse augmente significativement (environ 0.7 dex) sur l'intervalle t 5000 sondé entre -3.8 et -2.5. Ceci est la première détection de stratification du manganèse dans une étoile de type HgMn. Pour HD 175640, des indices de stratification du manganèse existent, mais l'étude d'autres spectres aiderait à confirmer ou non, la stratification de cet élément.

  18. Drug-enhanced carbon monoxide production from heme by cytochrome P450 reductase.

    PubMed

    Vukomanovic, Dragic; Rahman, Mona N; Jia, Zongchao; Nakatsu, Kanji

    2017-01-01

    Carbon monoxide (CO) formed endogenously is considered to be cytoprotective, and the vast majority of CO formation is attributed to the degradation of heme by heme oxygenases-1 and -2 (HO-1, HO-2). Previously, we observed that brain microsomes containing HO-2 produced many-fold more CO in the presence of menadione and its congeners; herein we explored these observations further. We determined the effects of various drugs on CO production of rat brain microsomes and recombinant human cytochrome P450 reductase (CPR); CO was measured by gas chromatography with reductive detection. Brain microsomes of Sprague-Dawley rats or recombinant human cytochrome P450 reductase (CPR) were incubated with NADPH and various drugs in closed vials in phosphate buffer at pH 7.4 and 37°C. After 15 minutes, the reaction was stopped by cooling in dry ice, and the headspace gas was analyzed for CO production using gas chromatography with reductive (mercuric oxide) detection. We observed drug-enhanced CO production in the presence of both microsomes and recombinant CPR alone; the presence of HO was not required. A range of structurally diverse drugs were capable of amplifying this CO formation; these molecules had structures consistent with redox cycling capability. The addition of catalase to a reaction mixture, that contained activating drugs, inhibited the production of CO. Drug-enhanced CO formation can be catalyzed by CPR. The mechanism of CPR activation was not through classical drug-receptor mediation. Redox cycling may be involved in the drug-induced amplification of CO production by CPR through the production of reactive oxygen species.

  19. Perchlorate Reductase Is Distinguished by Active Site Aromatic Gate Residues*

    PubMed Central

    Youngblut, Matthew D.; Tsai, Chi-Lin; Clark, Iain C.; Carlson, Hans K.; Maglaqui, Adrian P.; Gau-Pan, Phonchien S.; Redford, Steven A.; Wong, Alan; Tainer, John A.; Coates, John D.

    2016-01-01

    Perchlorate is an important ion on both Earth and Mars. Perchlorate reductase (PcrAB), a specialized member of the dimethylsulfoxide reductase superfamily, catalyzes the first step of microbial perchlorate respiration, but little is known about the biochemistry, specificity, structure, and mechanism of PcrAB. Here we characterize the biophysics and phylogeny of this enzyme and report the 1.86-Å resolution PcrAB complex crystal structure. Biochemical analysis revealed a relatively high perchlorate affinity (Km = 6 μm) and a characteristic substrate inhibition compared with the highly similar respiratory nitrate reductase NarGHI, which has a relatively much lower affinity for perchlorate (Km = 1.1 mm) and no substrate inhibition. Structural analysis of oxidized and reduced PcrAB with and without the substrate analog SeO32− bound to the active site identified key residues in the positively charged and funnel-shaped substrate access tunnel that gated substrate entrance and product release while trapping transiently produced chlorate. The structures suggest gating was associated with shifts of a Phe residue between open and closed conformations plus an Asp residue carboxylate shift between monodentate and bidentate coordination to the active site molybdenum atom. Taken together, structural and mutational analyses of gate residues suggest key roles of these gate residues for substrate entrance and product release. Our combined results provide the first detailed structural insight into the mechanism of biological perchlorate reduction, a critical component of the chlorine redox cycle on Earth. PMID:26940877

  20. Naegleria fowleri: a free-living highly pathogenic amoeba contains trypanothione/trypanothione reductase and glutathione/glutathione reductase systems.

    PubMed

    Ondarza, Raúl N; Hurtado, Gerardo; Tamayo, Elsa; Iturbe, Angélica; Hernández, Eva

    2006-11-01

    This paper presents definitive data showing that the thiol-bimane compound isolated and purified by HPLC from Naegleria fowleri trophozoites unequivocally corresponds by matrix assisted laser-desorption ionization-time-of-flight MS, to the characteristic monoprotonated ion of trypanothione-(bimane)(2) [M(+)H(+)] of m/z 1104.57 and to the trypanothione-(bimane) of m/z 914.46. The trypanothione disulfide T(S)(2) was also found to have a molecular ion of m/z 723.37. Additionally HPLC demonstrated that thiol-bimane compounds corresponding to cysteine and glutathione were present in Naegleria. The ion patterns of the thiol-bimane compounds prepared from commercial trypanothione standard, Entamoeba histolytica and Crithidia luciliae are identical to the Naegleria thiol-bimane compound. Partially purified extracts from N. fowleri showed the coexistence of glutathione and trypanothione reductases activities. There is not doubt that the thiol compound trypanothione, which was previously thought to occur only in Kinetoplastida, is also present in the human pathogens E. histolytica and N. fowleri, as well as in the non-pathogenic euglenozoan E. gracilis. The presence of the trypanothione/trypanothione reductase system in N. fowleri creates the possibility of using this enzyme as a new "drug target" for rationally designed drugs to eliminate the parasite, without affecting the human host.

  1. Nitrate Reductase Regulates Expression of Nitrite Uptake and Nitrite Reductase Activities in Chlamydomonas reinhardtii 1

    PubMed Central

    Galván, Aurora; Cárdenas, Jacobo; Fernández, Emilio

    1992-01-01

    In Chlamydomonas reinhardtii mutants defective at the structural locus for nitrate reductase (nit-1) or at loci for biosynthesis of the molybdopterin cofactor (nit-3, nit-4, or nit-5 and nit-6), both nitrite uptake and nitrite reductase activities were repressed in ammonium-grown cells and expressed at high amounts in nitrogen-free media or in media containing nitrate or nitrite. In contrast, wild-type cells required nitrate induction for expression of high levels of both activities. In mutants defective at the regulatory locus for nitrate reductase (nit-2), very low levels of nitrite uptake and nitrite reductase activities were expressed even in the presence of nitrate or nitrite. Both restoration of nitrate reductase activity in mutants defective at nit-1, nit-3, and nit-4 by isolating diploid strains among them and transformation of a structural mutant upon integration of the wild-type nit-1 gene gave rise to the wild-type expression pattern for nitrite uptake and nitrite reductase activities. Conversely, inactivation of nitrate reductase by tungstate treatment in nitrate, nitrite, or nitrogen-free media made wild-type cells respond like nitrate reductase-deficient mutants with respect to the expression of nitrite uptake and nitrite reductase activities. Our results indicate that nit-2 is a regulatory locus for both the nitrite uptake system and nitrite reductase, and that the nitrate reductase enzyme plays an important role in the regulation of the expression of both enzyme activities. PMID:16668656

  2. Volatilization of fluorescein mercuric acetate by marine bacterial from Minamata Bay

    SciTech Connect

    Nakamura, Kunihiko )

    1989-05-01

    Some bacteria that live in a mercury-polluted environment are resistant to mercury compounds. A majority of these mercury-resistant bacterial have been found to volatilize organic as well as inorganic mercury compounds into elemental mercury vapor by means of their enzymes. One compound, fluorescein mercuric acetate (FMA) has long been in use as a disinfectant in hospitals; yet, there has been little definitive information on bacterial resistance to this compound. Minamata Bay has been heavily polluted by mercury, which has caused methylmercury poisoning in humans, called Minamata disease. Sediments from the Bay still contain high concentrations of mercury. The percentage of mercury-resistant bacteria in the total bacterial count is higher in these sediments than in those of other marine environments. FMA-pollution, however, has not been reported. Research into the mechanism of bacterial resistance to FMA will not only add to our general understanding of the ability of certain bacteria to resist mercury, but will also help in defining the role bacteria play in the mercury cycle of a mercury-polluted environment. The purpose of the present study is to determine the mechanism of resistance to FMA of the FMA-resistant bacteria living in the Bay.

  3. Recent advances with mercuric iodide x-ray detectors and large-area silicon avalanche photodiodes

    NASA Astrophysics Data System (ADS)

    Dabrowski, Andrzej J.; Iwanczyk, Jan S.; Wang, Yuzhong J.; Madden, Michael C.; Szawlowski, Marek

    1993-10-01

    The paper presents advances in two sensor technologies: (1) Mercuric Iodide (HgI2) X-ray Detector Technology and, (2) Large Area Silicon Avalanche Photodiode (APD) Technology, which after years of development have recently produced commercially viable devices. Large Area Silicon Avalanche Photodiodes, which are solid-state light sensitive devices with internal amplification, combine the convenience, ruggedness and low power consumption of traditional semiconductor p-n and p-i-n photodiodes with the high light sensitivity and large photosensitive area approaching the active areas of traditional vacuum photomultiplier tubes. Device approaching 1-inch diameter with internal gain of up to 1000, have been developed by utilizing a beveled edge structure. By combining APD's with scintillation crystals, resolution of 6% (FWHM) was obtained for 662 keV energy line of 137Cs using a CsI(Tl) scintillator, and 7% (FWHM) was obtained using a NaI(Tl) scintillator. Resolution of 14% (FWHM) at room temperature and 11% (FWHM) at 0 degree(s)C have been obtained for APD's coupled to BGO scintillators. Rise times of 3 ns were measured by applying an impulse signal input, to a 200 mm2 device.

  4. Effect of mercuric chloride feeding on sexual maturity, egg production and fertility in Japanese quail

    USGS Publications Warehouse

    Hill, E.F.; Shaffner, C.S.

    1973-01-01

    Japanese quail (Coturnix c. japonica) were fed 0, 8, 16 or 32 p.p.m. of mercury as mercuric chloride from 3 days of age through 20 weeks of age. The onset of egg production generally occurred earlier for hens fed HgCl2. Average age in days at first oviposition for the control, 8 p.p.m., 16 p.p.m. and 32 p.p.m. was 48.4, 50.9, 46.9 and 44.0 respectively. The average rate of egg productivity from first oviposition to attainment of full growth (9 weeks of age) correlated positively with in increased dietary mercury (controls, 8 p.p.m., 16 p.p.m., 32 p.p.m. ? 75.2, 69.3, 86.1 and 93.3% respectively). By 20 weeks of age productivity was 81.0, 80.6, 87.5 and 92.9% for control, 8, 16 and 32 p.p.m. groups respectively. Fertility was depressed when hens were fed HgCl2. At 9 weeks of age average control fertility was 59% contrasted with 25% for the 32 p.p.m. group. At 12 weeks fertility increased to 89% and 57% for these groups. From this study it is apparent. that the onset and rate of egg production was stimulated by HgCl2, but fertility was adversely affected.

  5. Physical vapor transport of mercurous chloride under a nonlinear thermal profile

    NASA Technical Reports Server (NTRS)

    Mennetrier, Christophe; Duval, Walter M. B.; Singh, Narsingh B.

    1992-01-01

    Our study investigates numerically the flow field characteristics during the growth of mercurous chloride (Hg2Cl2) crystals in a rectangular ampoule under terrestrial and microgravity conditions for a nonlinear thermal gradient. With a residual gas lighter than the nutrient, the solutal Grashof number is dominant. We observe that in tilted configurations, when solutal convection is dominant, the maximum transport rate occurs at approximately 40 percent. For the vertical configurations, we were able to obtain solutions only for the cases either below the critical Rayleigh numbers or the stabilized configurations. The total mass flux decreases exponentially with an increase of pressure of residual gas, but it increases following a power law with the temperature difference driving the transport. The nonlinear thermal gradient appears to destabilize the flow field when thermal convection is dominant for both vertical top-heated and bottom-heated configurations. However, when the solutal Grashof number is dominant, the density gradient resulting from the solutal gradient appears to stabilize the flow for the bottom-heated configuration. The flow field for the top-heated configuration is destabilized for high Grashof numbers. The microgravity environment provides a means for lowering convection. For gravity levels of 10(exp -3) g(0) or less, the Stefan wind drives the flow, and no recirculating cell is predicted.

  6. Investigation of a mercurous chloride acousto-optic cell based on longitudinal acoustic mode.

    PubMed

    Gupta, Neelam

    2009-03-01

    A number of spectral imagers using acousto-optic tunable filters (AOTFs) operating from the UV to the longwave infrared (LWIR) using KDP, MgF(2), TeO(2), and Tl(3)AsSe(3) crystals to cover different spectral regions have been developed. In the LWIR there is a lack of high quality acousto-optic (AO) materials. Mercurous halide (Hg(2)Cl(2) and Hg(2)Br(2)) crystals are highly anisotropic with a high AO figure of merit due to slow acoustic velocities and high photoelastic constants and are transparent over a wide spectral region from 0.35 to 20 mum for Hg(2)Cl(2) and from 0.4 to 30 mum for Hg(2)Br(2). AO modulators, deflectors, and AOTFs based on these crystals can operate over a wide spectral range. Single crystals of these materials are being grown and some prototype devices have been fabricated. Results are presented from device characterization for an AO cell fabricated in Hg(2)Cl(2) based on longitudinal acoustic mode propagation. This device was very useful in demonstrating the AO interaction as well as soundness of the transducer bonding technique. Acoustic phase velocity is calculated and measured, diffraction efficiency is obtained from experiments, and the AO figure of merit of the sample is evaluated.

  7. Study of stoichiometry in mercuric iodide by low-temperature photoluminescence spectroscopy

    NASA Astrophysics Data System (ADS)

    Bao, Xue J.; James, Ralph B.; Hung, C.-Y.; Schlesinger, Tuviah E.; Cheng, A. Y.; Ortale, Carol; van den Berg, Lodewijk

    1993-02-01

    Low temperature (4.2 K) photoluminescence spectroscopy (PL) measurements were performed on mercuric iodide (HgI(subscript 2)) crystals that were surface-doped with either iodine or mercury. Two methods of treatment were used to achieve the surface doping. The first is the direct immersion of HgI(subscript 2) samples into potassium iodide (KI) aqueous solution saturated with iodine or immersion into elemental mercury liquid. The second is the storage of HgI(subscript 2) crystals under either iodine or mercury vapor. Certain features in the PL spectra were correlated with the stoichiometry of the HgI(subscript 2/ crystals modified by the surface doping. It was also found that if HgI(subscript 2) was exposed to air, an iodine deficient surface layer would form within a one-day period due to the preferential loss of iodine. Finally, the behavior of a broad emission band in the PL spectra and its implication in the fabrication of high quality HgI(subscript 2) nuclear detector is discussed.

  8. Development of a mercuric iodide detector array for in-vivo x-ray imaging

    SciTech Connect

    Patt, B.E.; Iwanczyk, J.S.; Tornai, M.P.; Levin, C.S.; Hoffman, E.J.

    1995-12-31

    A nineteen element mercuric iodide (HgI{sub 2}) detector array has been developed in order to investigate the potential of using this technology for in-vivo x-ray and gamma-ray imaging. A prototype cross-grid detector array was constructed with hexagonal pixels of 1.9 mm diameter (active area = 3.28 mm{sup 2}) and 0.2 mm thick septa. The overall detector active area is roughly 65 mm{sup 2}. A detector thickness of 1.2 mm was used to achieve about 100% efficiency at 60 keV and 67% efficiency at 140 keV The detector fabrication, geometry and structure were optimized for charge collection and to minimize crosstalk between elements. A section of a standard high resolution cast-lead gamma-camera collimator was incorporated into the detector to provide collimation matching the discrete pixel geometry. Measurements of spectral and spatial performance of the array were made using 241-Am and 99m-Tc sources. These measurements were compared with similar measurements made using an optimized single HgI{sub 2} x-ray detector with active area of about 3 mm{sup 2} and thickness of 500 {mu}m.

  9. Kinetics and mechanism of reaction between silver molybdate and mercuric iodide in solid state

    SciTech Connect

    Beg, M.A.; Rafiuddin

    1987-05-01

    The kinetics and the mechanism of the reaction between silver molybdate and mercuric iodide were studied in the solid state by X-ray, chemical analysis, and electrical conductivity measurements. This is a multistep reaction where Ag/sub 2/HgI/sub 4/ is formed as an intermediate. In an equimolar mixture of Ag/sub 2/MoO/sub 4/ and HgI/sub 2/, AgI an HgMoO/sub 4/ are formed, whereas in a 1:2 molar mixture Ag/sub 2/HgI/sub 4/ and HgMoO/sub 4/ are formed. The data for lateral diffusion best fit the equation X/sup n/ = kt, where X is the product thickness, t is time, and k and n are constants. This is a multistep solid state ionic reaction initiated by the diffusion of HgI/sub 2/ molecules as such and not through counterdiffusion of cations.

  10. Improved yield of high resolution mercuric iodide gamma-ray spectrometers

    SciTech Connect

    Gerrish, V.; van den Berg, L.

    1990-01-01

    Mercuric iodide (HgI{sub 2}) exhibits properties which make it attractive for use as a solid state nuclear radiation detector. The wide bandgap (E{sub g} = 2.1 eV) and low dark current allow room temperature operation, while the high atomic number provides a large gamma-ray cross section. However, poor hole transport has been a major limitation in the routine fabrication of high-resolution spectrometers using this material. This paper presents the results of gamma-ray response and charge transport parameter measurements conducted during the past year at EG G/EM on 96 HgI{sub 2} spectrometers. The gamma-ray response measurements reveal that detector quality is correlated with the starting material used in the crystal growth. In particular, an increased yield of high-resolution spectrometers was obtained from HgI{sub 2} which was synthesized by precipitation from an aqueous solution, as opposed to using material from commercial vendors. Data are also presented which suggest that better spectrometer performance is tied to improved hole transport. Finally, some initial results on a study of detector uniformity reveal spatial variations which may explain why the correlation between hole transport parameters and spectrometer performance is sometimes violated. 6 refs., 3 figs.

  11. Development of mercuric iodide energy dispersive x-ray array detectors

    SciTech Connect

    Iwanczyk, J.S.; Warburton, W.K.; Dabrowski, A.J.; Hedman, B.; Hodgson, K.O.; Patt, B.E.

    1988-02-01

    There are various areas of synchrotron radiation research particularly Extended X-Ray Absorption Fine Structure (EXAFS) on dilute solutions and anomalous scattering, which would strongly benefit from the availability of energy dispersive detector arrays with high energy resolution and good spatial resolution. The goal of this development project is to produce high energy resolution mercuric iodide (HgI/sub 2/) detector sub-modules, consisting of several elements. These sub-modules can later be grouped into larger arrays of 100-400 elements. A prototype 5 element HgI/sub 2/ array detector was constructed and tested. Dimensions of each element were 7.3 mm x 0.7 mm. An energy resolution of 335 eV (FWHM) for Mn0K..cap alpha.. at 5.9 keV has been measured. The novel fiber-optic pulsed light feedback has been introduced into the charge preamplifiers in order to minimize electronic crosstalk between channels.

  12. Mercuric chloride-induced protein-losing enteropathy (PLE) in brown Norway (BN) rats

    SciTech Connect

    Knoflach, P.; Weiser, M.M.; Albini, B.

    1986-03-05

    Prolonged exposure to low doses of mercuric chloride (MC) may induce immunologically mediated kidney disease in man and animals. Mercury compounds are of growing importance as environmental pollutants. Twenty female BN rats were gavaged with 150 microgram MC/100 gm body weight 3x/wk for up to 39 wks. Starting with wk 2, rat intestines demonstrated linear IgG and IgA deposits along the vascular and intestinal basement membranes (VBM and IBM). Serum antibodies to IBM were observed during the first 4 wks of gavage. At wk 11, first granular deposits of IgG and C3 were observed along VBM. Only after wk 35 were granular deposits also seen along the IBM. Using radioactive chromium chloride, 50% of rats with granular deposits along BM showed significantly increased protein loss into the intestines. Thus, granular deposits of IgG and C3 along the IBM, probably representing immune complexes, may lead to PLE. This animal model may contribute to the understanding of the pathogenesis of PLE in man described in graft-vrs-host reactions following bone marrow grafts, allergic enteritides, inflammatory bowel disease, and arsenic intoxication, as well as the assessment of biological effects of environmental pollutants.

  13. Detection of mercuric chloride by photofragment emission using a frequency-converted fiber amplifier

    SciTech Connect

    Hoops, A.A.; Reichardt, T.A.; Kliner, D.A.V.; Koplow, J.P.; Moore, S.W.

    2007-07-15

    A real-time, noninvasive approach for detecting trace amounts of vapor-phase mercuric chloride (HgCl{sub 2}) in combustion flue gas is demonstrated using a near-infrared pulsed fiber amplifier that is frequency converted to the ultraviolet. Excitation of the HgCl{sub 2} (1{Pi}1{sub u}{l_arrow} 1{Sigma}1g+) transition at 213 nm generates 253.7 nm emission from the Hg (6{sup 3}P{sub 1}) photoproduct that is proportional to the concentration of HgC1{sub 2}. A measured quadratic dependence of the HgCl{sub 2} photofragment emission (PFE) signal on the laser irradiance indicates that the photodissociation process involves two-photon excitation. Additionally, low concentrations of HgCl{sub 2} are detected with the PFE approach in an environment characteristic of coal-fired power-plant flue gas using this compact solid-state laser source. A detection limit of 0.7 ppb is extrapolated from these results.

  14. Intrarenal distribution of mercury in the rat: effect of administered dose of mercuric chloride

    SciTech Connect

    Zalups, R.K.; Diamond, G.L.

    1987-01-01

    The authors recently observed that the distribution of mercury in the hypertrophied remnant kidneys of uninephrectomized rats was different from that in the kidneys of sham-operated rats when given the same non-toxic dose of mercuric chloride (HgCl/sub 2/; 0.5 ..mu..mol/kg). These observations are quite significant, since the altered intrarenal distribution of mercury in uninephrectomized rats may cause uninephrectomized rats to develop more severe tubular necrosis in the outer medulla than sham-operated rats. In the experiments described above, the mercury burden of the hypertrophied remnant kidneys from the uninephrectomized rats was approximately twice that of each of the kidneys from the sham-operated rats. Thus, the altered intrarenal distribution of mercury in the uninephrectomized rats may be, in part, the result of the remnant kidney being exposed to more mercury. Implicit in this hypothesis is the idea that the manner in which the kidney accumulates mercury is dependent on the amount of mercury it is exposed to. If this is the case, then one would predict that the intrarenal accumulation of mercury in rats with two kidneys would change as the administered dose of HgCl/sub 2/ is increased from the dose of 0.5 ..mu..mol/kg. The principal aim of this study was to test this hypothesis.

  15. Mercuric iodide room-temperature array detectors for gamma-ray imaging

    SciTech Connect

    Patt, B.

    1994-11-15

    Significant progress has been made recently in the development of mercuric iodide detector arrays for gamma-ray imaging, making real the possibility of constructing high-performance small, light-weight, portable gamma-ray imaging systems. New techniques have been applied in detector fabrication and then low noise electronics which have produced pixel arrays with high-energy resolution, high spatial resolution, high gamma stopping efficiency. Measurements of the energy resolution capability have been made on a 19-element protypical array. Pixel energy resolutions of 2.98% fwhm and 3.88% fwhm were obtained at 59 keV (241-Am) and 140-keV (99m-Tc), respectively. The pixel spectra for a 14-element section of the data is shown together with the composition of the overlapped individual pixel spectra. These techniques are now being applied to fabricate much larger arrays with thousands of pixels. Extension of these principles to imaging scenarios involving gamma-ray energies up to several hundred keV is also possible. This would enable imaging of the 208 keV and 375-414 keV 239-Pu and 240-Pu structures, as well as the 186 keV line of 235-U.

  16. Mercury Disposition in Suckling Rats: Comparative Assessment Following Parenteral Exposure to Thiomersal and Mercuric Chloride

    PubMed Central

    Blanuša, Maja; Orct, Tatjana; Vihnanek Lazarus, Maja; Sekovanić, Ankica; Piasek, Martina

    2012-01-01

    Due to the facts that thiomersal-containing vaccine is still in use in many developing countries, and all forms of mercury have recognised neurotoxic, nephrotoxic, and other toxic effects, studies on disposition of ethylmercury and other mercury forms are still justified, especially at young age. Our investigation aimed at comparing mercury distribution and rate of excretion in the early period of life following exposure to either thiomersal (TM) or mercuric chloride (HgCl2) in suckling rats. Three experimental groups were studied: control, TM, and HgCl2, with 12 to18 pups in each. Both forms of mercury were administered subcutaneously in equimolar quantities (0.81 μmol/kg b.w.) three times during the suckling period (on the days of birth 7, 9, and 11) to mimic the vaccination regimen in infants. After the last administration of TM or HgCl2, total mercury retention and excretion was assessed during following six days. In TM-exposed group mercury retention was higher in the brain, enteral excretion was similar, and urinary excretion was much lower compared to HgCl2-exposed sucklings. More research is still needed to elucidate all aspects of toxicokinetics and most harmful neurotoxic potential of various forms of mercury, especially in the earliest period of life. PMID:22899883

  17. Human aldose reductase and human small intestine aldose reductase are efficient retinal reductases: consequences for retinoid metabolism.

    PubMed

    Crosas, Bernat; Hyndman, David J; Gallego, Oriol; Martras, Sílvia; Parés, Xavier; Flynn, T Geoffrey; Farrés, Jaume

    2003-08-01

    Aldo-keto reductases (AKRs) are NAD(P)H-dependent oxidoreductases that catalyse the reduction of a variety of carbonyl compounds, such as carbohydrates, aliphatic and aromatic aldehydes and steroids. We have studied the retinal reductase activity of human aldose reductase (AR), human small-intestine (HSI) AR and pig aldehyde reductase. Human AR and HSI AR were very efficient in the reduction of all- trans -, 9- cis - and 13- cis -retinal ( k (cat)/ K (m)=1100-10300 mM(-1).min(-1)), constituting the first cytosolic NADP(H)-dependent retinal reductases described in humans. Aldehyde reductase showed no activity with these retinal isomers. Glucose was a poor inhibitor ( K (i)=80 mM) of retinal reductase activity of human AR, whereas tolrestat, a classical AKR inhibitor used pharmacologically to treat diabetes, inhibited retinal reduction by human AR and HSI AR. All- trans -retinoic acid failed to inhibit both enzymes. In this paper we present the AKRs as an emergent superfamily of retinal-active enzymes, putatively involved in the regulation of retinoid biological activity through the assimilation of retinoids from beta-carotene and the control of retinal bioavailability.

  18. Neuroprotective role for carbonyl reductase?

    PubMed

    Maser, Edmund

    2006-02-24

    Oxidative stress is increasingly implicated in neurodegenerative disorders including Alzheimer's, Parkinson's, Huntington's, and Creutzfeld-Jakob diseases or amyotrophic lateral sclerosis. Reactive oxygen species seem to play a significant role in neuronal cell death in that they generate reactive aldehydes from membrane lipid peroxidation. Several neuronal diseases are associated with increased accumulation of abnormal protein adducts of reactive aldehydes, which mediate oxidative stress-linked pathological events, including cellular growth inhibition and apoptosis induction. Combining findings on neurodegeneration and oxidative stress in Drosophila with studies on the metabolic characteristics of the human enzyme carbonyl reductase (CR), it is clear now that CR has a potential physiological role for neuroprotection in humans. Several lines of evidence suggest that CR represents a significant pathway for the detoxification of reactive aldehydes derived from lipid peroxidation and that CR in humans is essential for neuronal cell survival and to confer protection against oxidative stress-induced brain degeneration.

  19. ADP-ribosylation of dinitrogenase reductase in Rhodobacter capsulatus

    SciTech Connect

    Jouanneau, Y.; Roby, C.; Meyer, C.M.; Vignais, P.M. )

    1989-07-25

    In the photosynthetic bacterium Rhodobacter capsulatus, nitrogenase is regulated by a reversible covalent modification of Fe protein or dinitrogenase reductase (Rc2). The linkage of the modifying group to inactive Rc2 was found to be sensitive to alkali and to neutral hydroxylamine. Complete release of the modifying group was achieved by incubation of inactive Rc2 in 0.4 or 1 M hydroxylamine. After hydroxylamine treatment of the Rc2 preparation, the modifying group could be isolated and purified by affinity chromatography and ion-exchange HPLC. The modifying group comigrated with ADP-ribose on both ion-exchange HPLC and thin-layer chromatography. Analyses by {sup 31}P NMR spectroscopy and mass spectrometry provided further evidence that the modifying group was ADP-ribose. The NMR spectrum of inactive Rc2 exhibited signals characteristic of ADP-ribose; integration of these signals allowed calculation of a molar ration ADP-ribose/Rc2 of 0.63. A hexapeptide carrying the ADP-ribose moiety was purified from a subtilisin digest of inactive Rc2. The structure of this peptide, determined by amino acid analysis and sequencing, is Gly-Arg(ADP-ribose)-Gly-Val-Ile-Thr. This structure allows identification of the binding site for ADP-ribose as Arg 101 of the polypeptide chain of Rc2. It is concluded that nitrogenase activity in R. capsulatus is regulated by reversible ADP-ribosylation of a specific arginyl residue of dinitrogenase reductase.

  20. Purification and characterization of an NADPH-cytochrome P450 (cytochrome c) reductase from spearmint (Mentha spicata) glandular trichomes.

    PubMed

    Ponnamperuma, K; Croteau, R

    1996-05-01

    Solubilized NADPH-cytochrome c (P450) reductase was purified to homogeneity from an extract of spearmint (Mentha spicata) glandular trichomes by dye-ligand interaction chromatography on Matrex-Gel Red A and affinity chromatography on 2', 5'-adenosine diphosphate agarose. SDS-PAGE of the purified enzyme preparation revealed the presence of two similar proteins with masses of 82 kDa (major) and 77 kDa (minor) that crossreacted on immunoblot analysis with polyclonal antibodies directed against NADPH-cytochrome P450 reductase from Jerusalem artichoke and from mung bean. Complete immunoinhibition of reductase activity was observed with both types of polyclonal antibodies, while only partial inhibition of activity resulted using a family of monoclonal antibodies directed against the Jerusalem artichoke cytochrome P450 reductase. Inhibition of the spearmint oil gland cytochrome c reductase was also observed with the diphenyliodonium ion. The K(m) values for the cosubstrates NADPH and cytochrome c were 6.2 and 3.7 microM, respectively, and the pH optimum for activity was at 8.5. The NADPH-cytochrome c reductase reconstituted NADPH-dependent (-)-4S-limonene-6-hydroxylase activity in the presence of cytochrome P450, purified from the microsomal fraction of spearmint oil gland cells and dilauroyl phosphatidyl choline. These characteristics establish the identity of the purified enzyme as a NADPH-cytochrome P450 reductase.

  1. Hepato- and nephroprotective effects of bradykinin potentiating factor from scorpion (Buthus occitanus) venom on mercuric chloride-treated rats.

    PubMed

    Salman, Muhammad M A; Kotb, Ahmed M; Haridy, Mohie A M; Hammad, Seddik

    2016-01-01

    Bioactive peptides such as bradykinin potentiating factor (BPF), have, anti-oxidative, anti-inflammatory, immunomodulatory and ameliorative effects in chronic diseases and play a potential role in cancer prevention. It is known that the liver and kidney accumulate inorganic mercury upon exposure, which often leads to mercury intoxication in these organs. In this study, we investigated the effect of bradykinin potentiating factor (BPF), a scorpion venom peptide, on mercuric chloride-induced hepatic and renal toxicity in rats. We used 20 adult male Albino rats divided into four equal groups: the first group was injected with saline (control); the second group was administered daily with mercuric chloride (HgCl2) for 2 weeks; the third group was administered with BPF twice weekly for 2 successive weeks, while the fourth group was exposed to BPF followed by HgCl2. We observed that HgCl2 treated rats had a significant increase in serum ALT, AST, ALP, creatinine and urea levels compared to control. Furthermore, HgCl2 treated rats showed a marked decrease in total proteins, albumin and uric acids compared to control. The previously studied parameters were not significantly changed in BPF pretreated rats compared to control. Moreover, a significant decrease in the activities of glutathione perioxidase (GSH), superoxide dismutase (SOD), and catalase (CAT), in addition to a significant increase in the level of malondialdehyde (MDA) were observed in hepatic and renal tissues of rats after HgCl2 treatment. In contrast, the HgCl2/BPF treated rats showed a significant elevation in the activity of GSH, SOD, and CAT accompanied with a significant regression in the level of MDA compared to the HgCl2 exposed rats. We conclude that treatment with BPF is a promising prophylactic approach for the management of mercuric chloride-induced hepato- and nephro-toxicities.

  2. Hepato- and nephroprotective effects of bradykinin potentiating factor from scorpion (Buthus occitanus) venom on mercuric chloride-treated rats

    PubMed Central

    Salman, Muhammad M. A.; Kotb, Ahmed M.; Haridy, Mohie A. M.; Hammad, Seddik

    2016-01-01

    Bioactive peptides such as bradykinin potentiating factor (BPF), have, anti-oxidative, anti-inflammatory, immunomodulatory and ameliorative effects in chronic diseases and play a potential role in cancer prevention. It is known that the liver and kidney accumulate inorganic mercury upon exposure, which often leads to mercury intoxication in these organs. In this study, we investigated the effect of bradykinin potentiating factor (BPF), a scorpion venom peptide, on mercuric chloride-induced hepatic and renal toxicity in rats. We used 20 adult male Albino rats divided into four equal groups: the first group was injected with saline (control); the second group was administered daily with mercuric chloride (HgCl2) for 2 weeks; the third group was administered with BPF twice weekly for 2 successive weeks, while the fourth group was exposed to BPF followed by HgCl2. We observed that HgCl2 treated rats had a significant increase in serum ALT, AST, ALP, creatinine and urea levels compared to control. Furthermore, HgCl2 treated rats showed a marked decrease in total proteins, albumin and uric acids compared to control. The previously studied parameters were not significantly changed in BPF pretreated rats compared to control. Moreover, a significant decrease in the activities of glutathione perioxidase (GSH), superoxide dismutase (SOD), and catalase (CAT), in addition to a significant increase in the level of malondialdehyde (MDA) were observed in hepatic and renal tissues of rats after HgCl2 treatment. In contrast, the HgCl2/BPF treated rats showed a significant elevation in the activity of GSH, SOD, and CAT accompanied with a significant regression in the level of MDA compared to the HgCl2 exposed rats. We conclude that treatment with BPF is a promising prophylactic approach for the management of mercuric chloride-induced hepato- and nephro-toxicities. PMID:28337111

  3. Potential mechanism of fibronectin deposits in acute renal failure induced by mercuric chloride.

    PubMed

    Saball, E; Salvarrey, M; Serra, E; Picó, G; Elías, M M

    2001-10-01

    Many glomerular diseases are associated with changes in the expression and distribution in the components of extracellular matrix. A remarkable feature in acute renal failure induced by mercuric chloride in rats was large fibronectin (Fn) deposits in kidneys 1 h post-HgCl2 injection (5 mg/kg body wt., s.c.). Our study examined some mechanisms as potential explanation of the early Fn deposits in mercuric chloride induced acute renal failure. Total tissue mRNA of livers and kidneys of control and treated rats were used in Northern blot to determine whether accumulation of Fn in kidney is associated with increases in the expression of this protein in the kidney and/or in the liver. Analysis of Fn levels by Western blot were also performed. Northern blot did not show significant difference between control and treated rats, while the abundance of polymerized-Fn in kidney tissue was increased 1 h and 5 h post HgCl2 injection. HgCl2 influence on Fn folding was studied in vitro to detect possible conformational changes that could altered its normal pattern of matrix assembly and/or binding to different ligands. In this context HgCl2 binding to Fn was measured following native tryptophan fluorescence of Fn in the presence of HgCl2 (0.5-250 mM). Binding parameters for the HgCl2-Fn complex formation were Kd = (1.6 +/- 0.2) 10(-4) M; n = 1 +/- 0.3, indicating a low apparent affinity and one type binding site. Thermal denaturation of Fn showed, between 30-60 degrees C, a soft reversible conformational change, while between 75-80 degrees C a highly and irreversible transition is produced suggesting a modification of the tertiary structure. HgCl2 abolished this transition. The kinetic of thermal unfolding of Fn was also measured and the effects observed due to HgCl2 presence reinforced the previous data. Finally, the effect of HgCl2 on Fn binding to denatured collagen (gelatin) was also measured as an index of the effect of this cation on biological properties of Fn. Fn binds

  4. Toxic effects of mercuric chloride, methylmercuric chloride, and Emisan 6 (an organic mercurial fungicide) on ovarian recrudescence in the catfish

    SciTech Connect

    Kirubagaran, R.; Joy, K.P.

    1988-12-01

    Mercurial toxicity in fishes has been focused mainly on tissue uptake and subcellular distribution, nephrotoxicity, development, hatching and survivability of young ones and teratology. Very few studies have been attempted to investigate Hg toxicity on gonadal activity of fishes throughout the breeding season. In a previous investigation the authors have studied the toxic effects of mercuric chloride (HgCl/sub 2/), methylmercuric chloride (CH/sub 3/HgCl) and emisan 6 (an alkoxyalkyl fungicide) on the survival and histology of the kidney of the catfish, Clarias batrachus. The present report deals with toxic effects of these mercurials on ovarian recrudescence in the catfish, an economically important species in the subcontinent.

  5. Structure of Physarum polycephalum cytochrome b5 reductase at 1.56 A resolution.

    PubMed

    Kim, Sangwoo; Suga, Michihiro; Ogasahara, Kyoko; Ikegami, Terumi; Minami, Yoshiko; Yubisui, Toshitsugu; Tsukihara, Tomitake

    2007-04-01

    Physarum polycephalum cytochrome b(5) reductase catalyzes the reduction of cytochrome b(5) by NADH. The structure of P. polycephalum cytochrome b(5) reductase was determined at a resolution of 1.56 A. The molecular structure was compared with that of human cytochrome b(5) reductase, which had previously been determined at 1.75 A resolution [Bando et al. (2004), Acta Cryst. D60, 1929-1934]. The high-resolution structure revealed conformational differences between the two enzymes in the adenosine moiety of the FAD, the lid region and the linker region. The structural properties of both proteins were inspected in terms of hydrogen bonding, ion pairs, accessible surface area and cavity volume. The differences in these structural properties between the two proteins were consistent with estimates of their thermostabilities obtained from differential scanning calorimetry data.

  6. Enhanced mercuric chloride adsorption onto sulfur-modified activated carbons derived from waste tires.

    PubMed

    Yuan, Chung-Shin; Wang, Guangzhi; Xue, Sheng-Han; Ie, Iau-Ren; Jen, Yi-Hsiu; Tsai, Hsieh-Hung; Chen, Wei-Jin

    2012-07-01

    A number of activated carbons derived from waste tires were further impregnated by gaseous elemental sulfur at temperatures of 400 and 650 degrees C, with a carbon and sulfur mass ratio of 1:3. The capabilities of sulfur diffusing into the micropores of the activated carbons were significantly different between 400 and 650 degrees C, resulting in obvious dissimilarities in the sulfur content of the activated carbons. The sulfur-impregnated activated carbons were examined for the adsorptive capacity of gas-phase mercuric chloride (HgC1) by thermogravimetric analysis (TGA). The analytical precision of TGA was up to 10(-6) g at the inlet HgCl2 concentrations of 100, 300, and 500 microg/m3, for an adsorption time of 3 hr and an adsorption temperature of 150 degrees C, simulating the flue gas emitted from municipal solid waste (MSW) incinerators. Experimental results showed that sulfur modification can slightly reduce the specific surface area of activated carbons. High-surface-area activated carbons after sulfur modification had abundant mesopores and micropores, whereas low-surface-area activated carbons had abundant macropores and mesopores. Sulfur molecules were evenly distributed on the surface of the inner pores after sulfur modification, and the sulfur content of the activated carbons increased from 2-2.5% to 5-11%. After sulfur modification, the adsorptive capacity of HgCl2 for high-surface-area sulfurized activated carbons reached 1.557 mg/g (22 times higher than the virgin activated carbons). The injection of activated carbons was followed by fabric filtration, which is commonly used to remove HgCl2 from MSW incinerators. The residence time of activated carbons collected in the fabric filter is commonly about 1 hr, but the time required to achieve equilibrium is less than 10 min. Consequently, it is worthwhile to compare the adsorption rates of HgCl2 in the time intervals of < 10 and 10-60 min.

  7. Exposure to mercuric chloride induces developmental damage, oxidative stress and immunotoxicity in zebrafish embryos-larvae.

    PubMed

    Zhang, Qun-Fang; Li, Ying-Wen; Liu, Zhi-Hao; Chen, Qi-Liang

    2016-12-01

    Mercury (Hg) is a widespread environmental pollutant that can produce severe negative effects on fish even at very low concentrations. However, the mechanisms underlying inorganic Hg-induced oxidative stress and immunotoxicity in the early development stage of fish still need to be clarified. In the present study, zebrafish (Danio rerio) embryos were exposed to different concentrations of Hg(2+) (0, 1, 4 and 16μg/L; added as mercuric chloride, HgCl2) from 2h post-fertilization (hpf) to 168hpf. Developmental parameters and total Hg accumulation were monitored during the exposure period, and antioxidant status and the mRNA expression of genes related to the innate immune system were examined at 168hpf. The results showed that increasing Hg(2+) concentration and time significantly increased total Hg accumulation in zebrafish embryos-larvae. Exposure to 16μg/L Hg(2+) caused developmental damage, including increased mortality and malformation, decreased body length, and delayed hatching period. Meanwhile, HgCl2 exposure (especially in the 16μg/L Hg(2+) group) induced oxidative stress affecting antioxidant enzyme (CAT, GST and GPX) activities, endogenous GSH and MDA contents, as well as the mRNA levels of genes (cat1, sod1, gstr, gpx1a, nrf2, keap1, hsp70 and mt) encoding antioxidant proteins. Moreover, the transcription levels of several representative genes (il-1β, il-8, il-10, tnfα2, lyz and c3) involved in innate immunity were up-regulated by HgCl2 exposure, suggesting that inorganic Hg had the potential to induce immunotoxicity. Taken together, the present study provides evidence that waterborne HgCl2 exposure can induce developmental impairment, oxidative stress and immunotoxicity in the early development stage of fish, which brings insights into the toxicity mechanisms of inorganic Hg in fish.

  8. Genetics Home Reference: 5-alpha reductase deficiency

    MedlinePlus

    ... About half of these individuals adopt a male gender role in adolescence or early adulthood. Related Information ... 1730-5. Citation on PubMed Cohen-Kettenis PT. Gender change in 46,XY persons with 5alpha-reductase- ...

  9. A dissimilatory nitrite reductase in Paracoccus halodenitrificans

    NASA Technical Reports Server (NTRS)

    Grant, M. A.; Hochstein, L. I.

    1984-01-01

    Paracoccus halodenitrificans produced a membrane-associated nitrite reductase. Spectrophotometric analysis showed it to be associated with a cd-cytochrome and located on the inner side of the cytoplasmic membrane. When supplied with nitrite, membrane preparations produced nitrous oxide and nitric oxide in different ratios depending on the electron donor employed. The nitrite reductase was maximally active at relatively low concentrations of sodium chloride and remained attached to the membranes at 100 mM sodium chloride.

  10. A dissimilatory nitrite reductase in Paracoccus halodenitrificans

    NASA Technical Reports Server (NTRS)

    Grant, M. A.; Hochstein, L. I.

    1984-01-01

    Paracoccus halodenitrificans produced a membrane-associated nitrite reductase. Spectrophotometric analysis showed it to be associated with a cd-cytochrome and located on the inner side of the cytoplasmic membrane. When supplied with nitrite, membrane preparations produced nitrous oxide and nitric oxide in different ratios depending on the electron donor employed. The nitrite reductase was maximally active at relatively low concentrations of sodium chloride and remained attached to the membranes at 100 mM sodium chloride.

  11. Thioredoxin Reductase and its Inhibitors

    PubMed Central

    Saccoccia, Fulvio; Angelucci, Francesco; Boumis, Giovanna; Carotti, Daniela; Desiato, Gianni; Miele, Adriana E; Bellelli, Andrea

    2014-01-01

    Thioredoxin plays a crucial role in a wide number of physiological processes, which span from reduction of nucleotides to deoxyriboucleotides to the detoxification from xenobiotics, oxidants and radicals. The redox function of Thioredoxin is critically dependent on the enzyme Thioredoxin NADPH Reductase (TrxR). In view of its indirect involvement in the above mentioned physio/pathological processes, inhibition of TrxR is an important clinical goal. As a general rule, the affinities and mechanisms of binding of TrxR inhibitors to the target enzyme are known with scarce precision and conflicting results abound in the literature. A relevant analysis of published results as well as the experimental procedures is therefore needed, also in view of the critical interest of TrxR inhibitors. We review the inhibitors of TrxR and related flavoreductases and the classical treatment of reversible, competitive, non competitive and uncompetitive inhibition with respect to TrxR, and in some cases we are able to reconcile contradictory results generated by oversimplified data analysis. PMID:24875642

  12. A mercuric iodide detector system for X-ray astronomy. II - Results from flight tests of a balloon borne instrument

    NASA Technical Reports Server (NTRS)

    Vallerga, J. V.; Vanderspek, R. K.; Ricker, G. R.

    1983-01-01

    To establish the expected sensitivity of a new hard X-ray telescope design, described by Ricker et al., an experiment was conducted to measure the background counting rate at balloon altitudes (40 km) of mercuric iodide, a room temperature solid state X-ray detector. The prototype detector consisted of two thin mercuric iodide (HgI2) detectors surrounded by a large bismuth germanate scintillator operated in anticoincidence. The bismuth germanate shield vetoed most of the background counting rate induced by atmospheric gamma-rays, neutrons and cosmic rays. A balloon-borne gondola containing a prototype detector assembly was designed, constructed and flown twice in the spring of 1982 from Palestine, TX. The second flight of this instrument established a differential background counting rate of 4.2 + or - 0.7 x 10 to the -5th counts/s sq cm keV over the energy range of 40-80 keV. This measurement was within 50 percent of the predicted value. The measured rate is about 5 times lower than previously achieved in shielded NaI/CsI or Ge systems operating in the same energy range.

  13. Plasma enzyme activities in coturnix quail fed graded doses of DDE, polychlorinated biphenyl, malathion, and mercuric chloride

    USGS Publications Warehouse

    Dieter, M.P.

    1974-01-01

    Male Coturnix quail (Coturnix coturnix japonica) were fed diets for 12 weeks containing graded levels of DDE, polychlorinated biphenyl (Aroclor 1254), malathion, and mercuric chloride. Birds were bled prior to exposure and at 2, 4 and 12 weeks, and the plasma used to measure the activities of creatine kinase, aspartate aminotransferase, cholinesterase, fructose-diphosphate aldolase, and lactate dehydrogenase. Abnormal activity of certain plasma enzymes was noted in birds after 2 and 4 weeks, but these changes were not proportional to dose or exposure time. At 12 weeks increases in each of the activities of plasma enzymes of birds fed organochlorines, and decreases in cholinesterase activity of birds fed malathion or mercuric chloride, were proportional to the log dose of the respective agents. In addition, the pattern of enzyme responses in the 4 experimental groups had changed, and was illustrative of the specific type of substance that had been fed. The data suggest that qualitative and quantitative identification of environmental contaminants in birds, and perhaps a variety of wild animals, may be possible by utilization of multiple plasma enzyme assays. Residue analyses after 12 weeks of feeding showed that DDE accumulated in carcasses and livers at concentrations up to 4-fold higher than those in the diets. In contrast residues of Aroclor 1254 attained in carcasses were identical to, and in livers one-half of, the concentration in the feed. Mercury did not accumulate as much in the tissues; residues attained were one-twentieth or less of those in the feed.

  14. A mercuric iodide detector system for X-ray astronomy. II - Results from flight tests of a balloon borne instrument

    NASA Technical Reports Server (NTRS)

    Vallerga, J. V.; Vanderspek, R. K.; Ricker, G. R.

    1983-01-01

    To establish the expected sensitivity of a new hard X-ray telescope design, described by Ricker et al., an experiment was conducted to measure the background counting rate at balloon altitudes (40 km) of mercuric iodide, a room temperature solid state X-ray detector. The prototype detector consisted of two thin mercuric iodide (HgI2) detectors surrounded by a large bismuth germanate scintillator operated in anticoincidence. The bismuth germanate shield vetoed most of the background counting rate induced by atmospheric gamma-rays, neutrons and cosmic rays. A balloon-borne gondola containing a prototype detector assembly was designed, constructed and flown twice in the spring of 1982 from Palestine, TX. The second flight of this instrument established a differential background counting rate of 4.2 + or - 0.7 x 10 to the -5th counts/s sq cm keV over the energy range of 40-80 keV. This measurement was within 50 percent of the predicted value. The measured rate is about 5 times lower than previously achieved in shielded NaI/CsI or Ge systems operating in the same energy range.

  15. A mercuric detector system for X-ray astronomy. 2. Results from flight tests of a balloon borne instrument

    NASA Technical Reports Server (NTRS)

    Vallerga, J.; Vanderspek, R. K.; Ricker, G. R.

    1982-01-01

    To establish the expected sensitivity of a new hard X-ray telescope design, an experiment was conducted to measure the background counting rate at balloon altitudes (40 km) of mercuric iodide, a room temperature solid state X-ray detector. The prototype detector consisted of two thin mercuric iodide (HgI2) detectors surrounded by a large bismuth germanate (Bi4Ge3O12) scintillator operated in anticoincidence. The bismuth germanate shield vetoed most of the background counting rate induced by atmospheric gamma-rays, neutrons and cosmic rays. A balloon-borne gondola containing a prototype detector assembly was designed, constructed and flown twice in the spring of 1982 from Palestine, Texas. The second flight of this instrument established a differential background counting rate of 4.2 O.7 x 10-5 counts/sec cm keV over the energy range of 40 to 80 keV. This measurement was within 50% of the predicted value. The measured rate is approx 5 times lower than previously achieved in shielded NaI/CsI or Ge systems operating in the same energy range. The prediction was based on a Monte Carlo simulation of the detector assembly in the radiation environment at float altitude.

  16. Perchlorate Reductase Is Distinguished by Active Site Aromatic Gate Residues.

    PubMed

    Youngblut, Matthew D; Tsai, Chi-Lin; Clark, Iain C; Carlson, Hans K; Maglaqui, Adrian P; Gau-Pan, Phonchien S; Redford, Steven A; Wong, Alan; Tainer, John A; Coates, John D

    2016-04-22

    Perchlorate is an important ion on both Earth and Mars. Perchlorate reductase (PcrAB), a specialized member of the dimethylsulfoxide reductase superfamily, catalyzes the first step of microbial perchlorate respiration, but little is known about the biochemistry, specificity, structure, and mechanism of PcrAB. Here we characterize the biophysics and phylogeny of this enzyme and report the 1.86-Å resolution PcrAB complex crystal structure. Biochemical analysis revealed a relatively high perchlorate affinity (Km = 6 μm) and a characteristic substrate inhibition compared with the highly similar respiratory nitrate reductase NarGHI, which has a relatively much lower affinity for perchlorate (Km = 1.1 mm) and no substrate inhibition. Structural analysis of oxidized and reduced PcrAB with and without the substrate analog SeO3 (2-) bound to the active site identified key residues in the positively charged and funnel-shaped substrate access tunnel that gated substrate entrance and product release while trapping transiently produced chlorate. The structures suggest gating was associated with shifts of a Phe residue between open and closed conformations plus an Asp residue carboxylate shift between monodentate and bidentate coordination to the active site molybdenum atom. Taken together, structural and mutational analyses of gate residues suggest key roles of these gate residues for substrate entrance and product release. Our combined results provide the first detailed structural insight into the mechanism of biological perchlorate reduction, a critical component of the chlorine redox cycle on Earth. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  17. Development of a mercuric iodide detector array for medical imaging applications

    NASA Astrophysics Data System (ADS)

    Patt, Bradley E.; Iwanczyk, Jan S.; Tornai, Martin P.; Levin, Craig S.; Hoffman, Edward J.

    1995-02-01

    A nineteen element mercuric iodide (HgI 2) detector array has been developed as a prototype for a larger (169 element) array, which is intended for use as an intra-operative gamma camera (IOGC). This work is motivated by the need for identifying and removing residual tumor cells after the removal of bulk tumor, while sparing normal tissue. Prior to surgery, a tumor seeking radiopharmaceutical is injected into the patient, and the IOGC is used to locate and map out the radioactivity. The IOGC can be used with commercially available radioisotopes such as 201Tl, 99mTc, and 123I which have low energy X- and gamma-rays. The use of HgI 2 detector arrays in this application facilitates construction of an imaging head that is very compact and has a high signal-to-noise ratio. The prototype detectors were configured as discrete pixel elements joined by fine wires into novel pseudo crossed-grid arrays to promote improved electric field distribution compared with previous designs, and to maximize the fill factor for the expected circular probe shape. Pixel dimensions are hexagonal with 1.5 mm and 1.9 mm diameters separated by 0.2 mm thick lead septa. The overall detectors are hexagonal with a diameter of ˜1 cm. The sensitive detector thickness is 1.2 mm, which corresponds to >99% efficiency at 59 keV and 67% efficiency at 140 keV. Row, column, and pixel spectra have been measured on the prototypical detector array. Energy resolution was found to vary with the width of the row/column coincidence window that was applied. With the low edge of the coincidence window at 30% below the photopeak, pixel energy resolutions of 2.98% and 3.88% FWHM were obtained on the best individual pixels at 59 keV ( 241Am) and 140 keV ( 99mTc), respectively. To characterize this array as an imaging device, the spatial response of the pixels was measured with stepped point sources. The spatial response corresponded well with the pixel geometry, indicating that the spatial resolution was determined

  18. The aldo-keto reductase superfamily homepage.

    PubMed

    Hyndman, David; Bauman, David R; Heredia, Vladi V; Penning, Trevor M

    2003-02-01

    The aldo-keto reductases (AKRs) are one of the three enzyme superfamilies that perform oxidoreduction on a wide variety of natural and foreign substrates. A systematic nomenclature for the AKR superfamily was adopted in 1996 and was updated in September 2000 (visit www.med.upenn.edu/akr). Investigators have been diligent in submitting sequences of functional proteins to the Web site. With the new additions, the superfamily contains 114 proteins expressed in prokaryotes and eukaryotes that are distributed over 14 families (AKR1-AKR14). The AKR1 family contains the aldose reductases, the aldehyde reductases, the hydroxysteroid dehydrogenases and steroid 5beta-reductases, and is the largest. Other families of interest include AKR6, which includes potassium channel beta-subunits, and AKR7 the aflatoxin aldehyde reductases. Two new families include AKR13 (yeast aldose reductase) and AKR14 (Escherichia coli aldehyde reductase). Crystal structures of many AKRs and their complexes with ligands are available in the PDB and accessible through the Web site. Each structure has the characteristic (alpha/beta)(8)-barrel motif of the superfamily, a conserved cofactor binding site and a catalytic tetrad, and variable loop structures that define substrate specificity. Although the majority of AKRs are monomeric proteins of about 320 amino acids in length, the AKR2, AKR6 and AKR7 family may form multimers. To expand the nomenclature to accommodate multimers, we recommend that the composition and stoichiometry be listed. For example, AKR7A1:AKR7A4 (1:3) would designate a tetramer of the composition indicated. The current nomenclature is recognized by the Human Genome Project (HUGO) and the Web site provides a link to genomic information including chromosomal localization, gene boundaries, human ESTs and SNPs and much more.

  19. Chicken muscle aldose reductase: purification, properties and relationship to other chicken aldo/keto reductases.

    PubMed

    Murphy, D G; Davidson, W S

    1986-01-01

    An enzyme that catalyzes the NADPH-dependent reduction of a wide range of aromatic and hydroxy-aliphatic aldehydes was purified from chicken breast muscle. This enzyme shares many properties with mammalian aldose reductases including molecular weight, relative substrate specificity, Michaelis constants, an inhibitor specificity. Therefore, it seems appropriate to call this enzyme an aldose reductase (EC 1.1.1.21). Chicken muscle aldose reductase appears to be kinetically identical to an aldose reductase that has been purified from chicken kidney (Hara et al., Eur. J. Biochem. 133, 207-214) and to hen muscle L-glycol dehydrogenase (Bernado et al., Biochim. biophys. Acta 659, 189-198). The association of this aldose reductase with muscular dystrophy in the chick is discussed.

  20. Mineral supplementation increases erythrose reductase activity in erythritol biosynthesis from glycerol by Yarrowia lipolytica.

    PubMed

    Tomaszewska, Ludwika; Rymowicz, Waldemar; Rywińska, Anita

    2014-03-01

    The aim of this study was to examine the impact of divalent copper, iron, manganese, and zinc ions on the production of erythritol from glycerol by Yarrowia lipolytica and their effect on the activity of erythrose reductase. No inhibitory effect of the examined minerals on yeast growth was observed in the study. Supplementation with MnSO4 · 7H2O (25 mg l(-1)) increased erythritol production by Y. lipolytica by 14.5%. In the bioreactor culture with manganese ion addition, 47.1 g l(-1) of erythritol was produced from 100.0 g l(-1) of glycerol, which corresponded to volumetric productivity of 0.87 g l(-1) h(-1). The addition of Mn(2+) enhanced the intracellular activity of erythrose reductase up to 24.9 U g(-1) of dry weight of biomass (DW), hence, about 1.3 times more than in the control.

  1. Respiratory arsenate reductase as a bidirectional enzyme

    SciTech Connect

    Richey, Christine; Chovanec, Peter; Hoeft, Shelley E.; Oremland, Ronald S.; Basu, Partha; Stolz, John F.

    2009-05-01

    The haloalkaliphilic bacterium Alkalilimnicola ehrlichii is capable of anaerobic chemolithoautotrophic growth by coupling the oxidation of arsenite (As(III)) to the reduction of nitrate and carbon dioxide. Analysis of its complete genome indicates that it lacks a conventional arsenite oxidase (Aox), but instead possesses two operons that each encode a putative respiratory arsenate reductase (Arr). Here we show that one homolog is expressed under chemolithoautotrophic conditions and exhibits both arsenite oxidase and arsenate reductase activity. We also demonstrate that Arr from two arsenate respiring bacteria, Alkaliphilus oremlandii and Shewanella sp. strain ANA-3, is also biochemically reversible. Thus Arr can function as a reductase or oxidase. Its physiological role in a specific organism, however, may depend on the electron potentials of the molybdenum center and [Fe-S] clusters, additional subunits, or constitution of the electron transfer chain. This versatility further underscores the ubiquity and antiquity of microbial arsenic metabolism.

  2. Respiratory arsenate reductase as a bidirectional enzyme

    USGS Publications Warehouse

    Richey, C.; Chovanec, P.; Hoeft, S.E.; Oremland, R.S.; Basu, P.; Stolz, J.F.

    2009-01-01

    The haloalkaliphilic bacterium Alkalilimnicola ehrlichii is capable of anaerobic chemolithoautotrophic growth by coupling the oxidation of arsenite (As(III)) to the reduction of nitrate and carbon dioxide. Analysis of its complete genome indicates that it lacks a conventional arsenite oxidase (Aox), but instead possesses two operons that each encode a putative respiratory arsenate reductase (Arr). Here we show that one homolog is expressed under chemolithoautotrophic conditions and exhibits both arsenite oxidase and arsenate reductase activity. We also demonstrate that Arr from two arsenate respiring bacteria, Alkaliphilus oremlandii and Shewanella sp. strain ANA-3, is also biochemically reversible. Thus Arr can function as a reductase or oxidase. Its physiological role in a specific organism, however, may depend on the electron potentials of the molybdenum center and [Fe–S] clusters, additional subunits, or constitution of the electron transfer chain. This versatility further underscores the ubiquity and antiquity of microbial arsenic metabolism.

  3. Low energy X-ray spectra measured with a mercuric iodide energy dispersive spectrometer in a scanning electron microscope

    NASA Technical Reports Server (NTRS)

    Iwanczyk, J. S.; Dabrowski, A. J.; Huth, G. C.; Bradley, J. G.; Conley, J. M.

    1986-01-01

    A mercuric iodide energy dispersive X-ray spectrometer, with Peltier cooling provided for the detector and input field effect transistor, has been developed and tested in a scanning electron microscope. X-ray spectra were obtained with the 15 keV electron beam. An energy resolution of 225 eV (FWHM) for Mn-K(alpha) at 5.9 keV and 195 eV (FWHM) for the Mg-K line at 1.25 keV has been measured. Overall system noise level was 175 eV (FWHM). The detector system characterization with a carbon target demonstrated good energy sensitivity at low energies and lack of significant spectral artifacts at higher energies.

  4. A mercuric ensemble based on a cycloruthenated complex as a visual probe for iodide in aqueous solution

    NASA Astrophysics Data System (ADS)

    Su, Xianlong; Guo, Lieping; Ma, Yajuan; Li, Xianghong

    2016-01-01

    A new water-soluble cycloruthenated complex Ru(bthiq)(dcbpy)2+ (1, Hbthiq = 1-(2-benzo[b]thiophenyl)isoquinoline, dcbpy = 4,4‧-dicarboxylate-2,2‧-bipyridine) was designed and synthesized to form its mercuric ensemble (1-Hg2+) to achieve visual detection of iodide anions. The binding constant of 1-Hg2+ is calculated to be 2.40 × 104 M-1, which is lower than that of HgI2. Therefore, the addition of I- to the aqueous solution of 1-Hg2+lead to significant color changes from yellow to deep-red by the release of 1. The results showed that iodide anions could be easily detected by the naked eyes. The detection limit of iodide anion is calculated as 0.77 μM. In addition, an easily-prepared test strip of 1-Hg2+ was obtained successfully to detect iodide anions.

  5. A mercuric iodide detector system for X-ray astronomy. I - Design considerations and predictions of background and sensitivity

    NASA Technical Reports Server (NTRS)

    Ricker, G. R.; Vallerga, J. V.; Wood, D. R.

    1983-01-01

    Since the discovery of Sco X-1 initiated X-ray astronomy in 1962, this science has progressed in connection with the placement of X-ray photon detectors above the atmosphere by means of rockets, balloons, and satellites. In the last few years, studies have been conducted regarding the use of mercuric iodide (HgI2) as room temperature X-ray detector for applications in hard X-ray astronomy. These detectors combine a high quantum efficiency with good energy resolution. The sensitivity of an astronomical X-ray telescope is discussed, and a description is presented of a specific design accepted for the HDXT to be flown on Spacelab. Attention is given to predictions of the background counting rate of the detector assembly in this design, taking into account the results of a Monte Carlo simulation of the detector assembly in the radiation environment at balloon altitudes (40 km).

  6. In vitro interaction of selected phospholipid species with mercuric chloride using Fourier transform sup 1 H-NMR

    SciTech Connect

    Shinada, Masayuki; Muto, Hajime; Takizawa, Yukio )

    1991-09-01

    Many studies on the mercury toxicities have been accumulated since the outbreak of Minamata Disease.' There have been few reports on the reaction mechanisms of mercurials with phospholipids which substantially locate in biological membranes, although the interactions of nucleotides or nucleosides with mercurials have been reported. Recently, the study on the interaction of mercuric chloride (HgCl{sub 2}) with amino polar heads of model membranes containing phosphatidylserine (PS) and phosphatidylethanolamine (PE) has been reported, as the results from the fluorescence polarization analysis using 1,6-diphenyl-1,3,5-hexatriene. The authors demonstrate here the interactions of dioleoylphosphatidylethanolamine (DOPE) and dioleoylphosphatidylcholine (DOPC) with HgCl{sub 2}, using Fourier transform {sup 1}H-NMR ({sup 1}H-FT-NMR).

  7. Low energy X-ray spectra measured with a mercuric iodide energy dispersive spectrometer in a scanning electron microscope

    NASA Technical Reports Server (NTRS)

    Iwanczyk, J. S.; Dabrowski, A. J.; Huth, G. C.; Bradley, J. G.; Conley, J. M.

    1986-01-01

    A mercuric iodide energy dispersive X-ray spectrometer, with Peltier cooling provided for the detector and input field effect transistor, has been developed and tested in a scanning electron microscope. X-ray spectra were obtained with the 15 keV electron beam. An energy resolution of 225 eV (FWHM) for Mn-K(alpha) at 5.9 keV and 195 eV (FWHM) for the Mg-K line at 1.25 keV has been measured. Overall system noise level was 175 eV (FWHM). The detector system characterization with a carbon target demonstrated good energy sensitivity at low energies and lack of significant spectral artifacts at higher energies.

  8. Mitochondrial viability and apoptosis induced by aluminum, mercuric mercury and methylmercury in cell lines of neural origin.

    PubMed

    Toimela, Tarja; Tähti, Hanna

    2004-10-01

    Mercury and aluminum are considered to be neurotoxic metals, and they are often connected with the onset of neurodegenerative diseases. In this study, mercuric mercury, methylmercury and aluminum were studied in three different cell lines of neural origin. To evaluate the effects, mitochondrial cytotoxicity and apoptosis induced by the metals were measured after various incubation times. SH-SY5Y neuroblastoma, U 373MG glioblastoma, and RPE D407 retinal pigment epithelial cells were subcultured to appropriate cell culture plates and 0.01-1,000 microM concentrations of methylmercury, mercuric and aluminum chloride were added into the growth medium. In the assay measuring the mitochondrial dehydrogenase activity, WST-1, the cultures were exposed for 15 min, 24 or 48 h before measurement. Cells were allowed to recover from the exposure in part of the study. Apoptosis induced by the metals was measured after 6-, 24- and 48-h exposure times with the determination of activated caspase 3 enzyme. Mitochondrial assays showed a clear dose-response and exposure time-response to the metals. The most toxic was methylmercury (EC50 ~0.8 microM, 48 h), and the most sensitive cell line was the neuroblastoma cell line SH-SY5Y. Furthermore, there was marked mitochondrial activation, especially in connection with aluminum and methylmercury at low concentrations. This activation may be important during the initiation of cellular processes. All the metals tested induced apoptosis, but with a different time-course and cell-line specificity. In microscopic photographs, glioblastoma cells formed fibrillary tangles, and neuroblastoma cells settled along the fibrilles in cocultures of glial and neuronal cell lines during aluminum exposure. The study emphasized the toxicity of methylmercury to neural cells and showed that aluminum alters various cellular activities.

  9. Evaluation of nitrate reductase activity in Rhizobium japonicum

    SciTech Connect

    Streeter, J.G.; DeVine, P.J.

    1983-08-01

    Nitrate reductase activity was evaluated by four approaches, using four strains of Rhizobium japonicum and 11 chlorate-resistant mutants of the four strains. It was concluded that in vitro assays with bacteria or bacteroids provide the most simple and reliable assessment of the presence or absence of nitrate reductase. Nitrite reductase activity with methyl viologen and dithionite was found, but the enzyme activity does not confound the assay of nitrate reductase. 18 references

  10. Isolation, sequence identification and tissue expression profile of two novel soybean (glycine max) genes-vestitone reductase and chalcone reductase.

    PubMed

    Liu, G Y

    2009-09-01

    The complete mRNA sequences of two soybean (glycine max) genes-vestitone reductase and chalcone reductase, were amplified using the rapid amplification of cDNA ends methods. The sequence analysis of these two genes revealed that soybean vestitone reductase gene encodes a protein of 327 amino acids which has high homology with the vestitone reductase of Medicago sativa (77%). The soybean chalcone reductase gene encodes a protein of 314 amino acids that has high homology with the chalcone reductase of kudzu vine (88%) and medicago sativa (83%). The expression profiles of the soybean vestitone reductase and chalcone reductase genes were studied and the results indicated that these two soybean genes were differentially expressed in detected soybean tissues including leaves, stems, roots, inflorescences, embryos and endosperm. Our experiment established the foundation for further research on these two soybean genes.

  11. Inhibition of pea ferredoxin-NADP(H) reductase by Zn-ferrocyanide.

    PubMed

    Dupuy, Daniela L Catalano; Rial, Daniela V; Ceccarelli, Eduardo A

    2004-11-01

    Ferredoxin-NADP(H) reductases (FNRs) represent a prototype of enzymes involved in numerous metabolic pathways. We found that pea FNR ferricyanide diaphorase activity was inhibited by Zn2+ (Ki 1.57 microM). Dichlorophenolindophenol diaphorase activity was also inhibited by Zn2+ (Ki 1.80 microM), but the addition of ferrocyanide was required, indicating that the inhibitor is an arrangement of both ions. Escherichia coli FNR was also inhibited by Zn-ferrocyanide, suggesting that inhibition is a consequence of common structural features of these flavoenzymes. The inhibitor behaves in a noncompetitive manner for NADPH and for artificial electron acceptors. Analysis of the oxidation state of the flavin during catalysis in the presence of the inhibitor suggests that the electron-transfer process between NADPH and the flavin is not significantly altered, and that the transfer between the flavin and the second substrate is mainly affected. Zn-ferrocyanide interacts with the reductase, probably increasing the accessibility of the prosthetic group to the solvent. Ferredoxin reduction was also inhibited by Zn-ferrocyanide in a noncompetitive manner, but the observed Ki was about nine times higher than those for the diaphorase reactions. The electron transfer to Anabaena flavodoxin was not affected by Zn-ferrocyanide. Binding of the apoflavodoxin to the reductase was sufficient to overcome the inhibition by Zn-ferrocyanide, suggesting that the interaction of FNRs with their proteinaceous electron partners may induce a conformational change in the reductase that alters or completely prevents the inhibitory effect.

  12. Fumarate-Mediated Inhibition of Erythrose Reductase, a Key Enzyme for Erythritol Production by Torula corallina

    PubMed Central

    Lee, Jung-Kul; Koo, Bong-Seong; Kim, Sang-Yong

    2002-01-01

    Torula corallina, a strain presently being used for the industrial production of erythritol, has the highest erythritol yield ever reported for an erythritol-producing microorganism. The increased production of erythritol by Torula corallina with trace elements such as Cu2+ has been thoroughly reported, but the mechanism by which Cu2+ increases the production of erythritol has not been studied. This study demonstrated that supplemental Cu2+ enhanced the production of erythritol, while it significantly decreased the production of a major by-product that accumulates during erythritol fermentation, which was identified as fumarate by instrumental analyses. Erythrose reductase, a key enzyme that converts erythrose to erythritol in T. corallina, was purified to homogeneity by chromatographic methods, including ion-exchange and affinity chromatography. In vitro, purified erythrose reductase was significantly inhibited noncompetitively by increasing the fumarate concentration. In contrast, the enzyme activity remained almost constant regardless of Cu2+ concentration. This suggests that supplemental Cu2+ reduced the production of fumarate, a strong inhibitor of erythrose reductase, which led to less inhibition of erythrose reductase and a high yield of erythritol. This is the first report that suggests catabolite repression by a tricarboxylic acid cycle intermediate in T. corallina. PMID:12200310

  13. The anaerobic ribonucleoside triphosphate reductase from Escherichia coli requires S-adenosylmethionine as a cofactor.

    PubMed Central

    Eliasson, R; Fontecave, M; Jörnvall, H; Krook, M; Pontis, E; Reichard, P

    1990-01-01

    Extracts from anaerobically grown Escherichia coli contain an oxygen-sensitive activity that reduces CTP to dCTP in the presence of NADPH, dithiothreitol, Mg2+ ions, and ATP, different from the aerobic ribonucleoside diphosphate reductase (2'-deoxyribonucleoside-diphosphate: oxidized-thioredoxin 2'-oxidoreductase, EC 1.17.4.1) present in aerobically grown E. coli. After fractionation, the activity required at least five components, two heat-labile protein fractions and several low molecular weight fractions. One protein fraction, suggested to represent the actual ribonucleoside triphosphate reductase was purified extensively and on denaturing gel electrophoresis gave rise to several defined protein bands, all of which were stained by a polyclonal antibody against one of the two subunits (protein B1) of the aerobic reductase but not by monoclonal anti-B1 antibodies. Peptide mapping and sequence analyses revealed partly common structures between two types of protein bands but also suggested the presence of an additional component. Obviously, the preparations are heterogeneous and the structure of the reductase is not yet established. The second, crude protein fraction is believed to contain several ancillary enzymes required for the reaction. One of the low molecular weight components is S-adenosylmethionine; a second component is a loosely bound metal. We propose that S-adenosylmethionine together with a metal participates in the generation of the radical required for the reduction of carbon 2' of the ribosyl moiety of CTP. Images PMID:2185465

  14. Structure of Physarum polycephalum cytochrome b{sub 5} reductase at 1.56 Å resolution

    SciTech Connect

    Kim, Sangwoo; Suga, Michihiro; Ogasahara, Kyoko; Ikegami, Terumi; Minami, Yoshiko; Yubisui, Toshitsugu; Tsukihara, Tomitake

    2007-04-01

    The structure of P. polycephalum cytochrome b{sub 5} reductase, an enzyme which catalyzes the reduction of cytochrome b{sub 5} by NADH, was determined at a resolution of 1.56 Å. Physarum polycephalum cytochrome b{sub 5} reductase catalyzes the reduction of cytochrome b{sub 5} by NADH. The structure of P. polycephalum cytochrome b{sub 5} reductase was determined at a resolution of 1.56 Å. The molecular structure was compared with that of human cytochrome b{sub 5} reductase, which had previously been determined at 1.75 Å resolution [Bando et al. (2004 ▶), Acta Cryst. D60, 1929–1934]. The high-resolution structure revealed conformational differences between the two enzymes in the adenosine moiety of the FAD, the lid region and the linker region. The structural properties of both proteins were inspected in terms of hydrogen bonding, ion pairs, accessible surface area and cavity volume. The differences in these structural properties between the two proteins were consistent with estimates of their thermostabilities obtained from differential scanning calorimetry data.

  15. Fumarate Reductase Activity of Streptococcus faecalis

    PubMed Central

    Aue, B. J.; Diebel, R. H.

    1967-01-01

    Some characteristics of a fumarate reductase from Streptococcus faecalis are described. The enzyme had a pH optimum of 7.4; optimal activity was observed when the ionic strength of the phosphate buffer was adjusted to 0.088. The Km value of the enzyme for reduced flavin mononucleotide was 2 × 10−4 m as determined with a 26-fold preparation. In addition to fumarate, the enzyme reduced maleate and mesaconate. No succinate dehydrogenase activity was detected, but succinate did act as an inhibitor of the fumarate reductase activity. Other inhibitors were malonate, citraconate, and trans-, trans-muconate. Metal-chelating agents did not inhibit the enzyme. A limited inhibition by sulfhydryl-binding agents was observed, and the preparations were sensitive to air oxidation and storage. Glycine, alanine, histidine, and possibly lysine stimulated fumarate reductase activity in the cell-free extracts. However, growth in media supplemented with glycine did not enhance fumarate reductase activity. The enzymatic activity appears to be constitutive. PMID:4960892

  16. Post-translational Regulation of Nitrate Reductase

    USDA-ARS?s Scientific Manuscript database

    Nitrate reductase (NR) catalyzes the reduction of nitrate to nitrite, which is the first step in the nitrate assimilation pathway, but can also reduce nitrite to nitric oxide (NO), an important signaling molecule that is thought to mediate a wide array of of developmental and physiological processes...

  17. Construction and characterization of nitrate reductase-based amperometric electrode and nitrate assay of fertilizers and drinking water.

    PubMed

    Glazier, S A; Campbell, E R; Campbell, W H

    1998-04-15

    The construction and characterization of a nitrate reductase-based amperometric electrode for determination of nitrate ion is described. The electrode consisted of nitrate reductase held by dialysis membrane onto a Nafion-coated glassy carbon electrode. Methyl viologen was allowed to absorb into the Nafion layer, which acted as a reservoir for the electron mediator. The utility of the electrode to assay fertilizer and water sample for nitrate was demonstrated. The assays conducted with this electrode compared well with colorimetric and potentiometric assays of the same samples.

  18. Synthesis of symmetric disulfides as potential alternative substrates for trypanothione reductase and glutathione reductase: Part 1.

    PubMed

    Jaouhari, R; Besheya, T; McKie, J H; Douglas, K T

    1995-12-01

    The synthesis of a series of symmetrical disulfides as potential substrates of trypanothione reductase and glutathione reductase was described. The key intermediate in the synthetic approach was the choice of S-(t)butylmercapto-L-cysteine (1). The spermidine ring in the native substrate, trypanothione disulfide (TSST), was replaced with 3-dimethyl-aminopropylamine (DMAPA), while theγ-Glu moiety was replaced by phenylalanyl or tryptophanyl residues. The same modifications in theγ-Glu moiety of glutathione disulfide (GSSG) were applied.

  19. Control of dihydrofolate reductase messenger ribonucleic acid production

    SciTech Connect

    Leys, E.J.; Kellems, R.E.

    1981-11-01

    The authors used methotrexate-resistant mouse cells in which dihydrofolate reductase levels are approximately 500 times normal to study the effect of growth stimulation on dihydrofolate reductase gene expression. As a result of growth stimulation, the relative rate of dihydrofolate reductase protein synthesis increased threefold, reaching a maximum between 25 and 30 h after stimulation. The relative rate of dihydrofolate reductase messenger ribonucleic acid production (i.e., the appearance of dihydrofolate reductase messenger ribonucleic acid in the cytoplasm) increased threefold after growth stimulation and was accompanied by a corresponding increase in the relative steady-state level of dihydrofolate reductase ribonucleic acid in the nucleus. However, the increase in the nuclear level of dihydrofolate reductase ribonucleic acid was not accompanied by a significant increase in the relative rate of transcription of the dihydrofolate reductase genes. These data indicated that the relative rate of appearance of dihydrofolate reductase messenger ribonucleic acid in the cytoplasm depends on the relative stability of the dihydrofolate reductase ribonucleic acid sequences in the nucleus and is not dependent on the relative rate of transcription of the dihydrofolate reductase genes.

  20. Augmentation of CFTR maturation by S-nitrosoglutathione reductase

    PubMed Central

    Sawczak, Victoria; Zaidi, Atiya; Butler, Maya; Bennett, Deric; Getsy, Paulina; Zeinomar, Maryam; Greenberg, Zivi; Forbes, Michael; Rehman, Shagufta; Jyothikumar, Vinod; DeRonde, Kim; Sattar, Abdus; Smith, Laura; Corey, Deborah; Straub, Adam; Sun, Fei; Palmer, Lisa; Periasamy, Ammasi; Randell, Scott; Kelley, Thomas J.; Lewis, Stephen J.

    2015-01-01

    S-nitrosoglutathione (GSNO) reductase regulates novel endogenous S-nitrosothiol signaling pathways, and mice deficient in GSNO reductase are protected from airways hyperreactivity. S-nitrosothiols are present in the airway, and patients with cystic fibrosis (CF) tend to have low S-nitrosothiol levels that may be attributed to upregulation of GSNO reductase activity. The present study demonstrates that 1) GSNO reductase activity is increased in the cystic fibrosis bronchial epithelial (CFBE41o−) cells expressing mutant F508del-cystic fibrosis transmembrane regulator (CFTR) compared with the wild-type CFBE41o− cells, 2) GSNO reductase expression level is increased in the primary human bronchial epithelial cells expressing mutant F508del-CFTR compared with the wild-type cells, 3) GSNO reductase colocalizes with cochaperone Hsp70/Hsp90 organizing protein (Hop; Stip1) in human airway epithelial cells, 4) GSNO reductase knockdown with siRNA increases the expression and maturation of CFTR and decreases Stip1 expression in human airway epithelial cells, 5) increased levels of GSNO reductase cause a decrease in maturation of CFTR, and 6) a GSNO reductase inhibitor effectively reverses the effects of GSNO reductase on CFTR maturation. These studies provide a novel approach to define the subcellular location of the interactions between Stip1 and GSNO reductase and the role of S-nitrosothiols in these interactions. PMID:26637637

  1. Ferric and cupric reductase activities by iron-limited cells of the green alga Chlorella kessleri: quantification via oxygen electrode.

    PubMed

    Weger, Harold G; Walker, Crystal N; Fink, Michael B

    2007-10-01

    + concentrations with chelators other than desferrioxamine mesylate. With respect to cupric reductase activity, the O2 electrode consistently provided much higher estimates; we suggest that this was as a result of Cu2+ chelation by BCDS leading to a large underestimation of the true cupric reduction rate. These results suggest that an O2-electrode-based metal reductase assay system has some specific advantages compared with the traditional colorimetric assay system, including especially the ability to discriminate between the reduction of free metal ions and chelated metal ions.

  2. A genetic screen reveals a periplasmic copper chaperone required for nitrite reductase activity in pathogenic Neisseria.

    PubMed

    Jen, Freda E-C; Djoko, Karrera Y; Bent, Stephen J; Day, Christopher J; McEwan, Alastair G; Jennings, Michael P

    2015-09-01

    Under conditions of low oxygen availability, Neisseria meningitidis and Neisseria gonorrhoeae are able to respire via a partial denitrification pathway in which nitrite is converted to nitrous oxide. In this process, nitrite reductase (AniA), a copper (Cu)-containing protein converts nitrite to NO, and this product is converted to nitrous oxide by nitric oxide reductase (NorB). NorB also confers protection against toxic NO, and so we devised a conditional lethal screen, using a norB mutant, to identify mutants that were resistant to nitrite-dependent killing. After random-deletion mutagenesis of N. meningitidis, this genetic screen identified a gene encoding a Cu chaperone that is essential for AniA function, AccA. Purified AccA binds one Cu (I) ion and also possesses a second binding site for Cu (II). This novel periplasmic Cu chaperone (AccA) appears to be essential for provision of Cu ions to AniA of pathogenic Neisseria to generate an active nitrite reductase. Apart from the Neisseria genus, AccA is distributed across a wide range of environmental Proteobacteria species. © FASEB.

  3. Superoxide production during reduction of molecular oxygen by assimilatory nitrate reductase.

    PubMed

    Barber, M J; Kay, C J

    1996-02-15

    Assimilatory NADH:nitrate reductase catalyzes the transfer of reducing equivalents from NADH to molecular oxygen. Initial rate studies performed under conditions of optimal pH (8.0) and constant ionic strength (mu = 0.2) revealed that the maximal rate of activity with molecular oxygen was 0.5% (0.44 mumol NADH consumed/min/nmol heme) with a Km for O2 of 586 microM. NADH:molecular oxygen reductase activity exhibited a pH optimum of 9.2, was inhibited by cyanide, and was unaffected by changes in ionic strength or the presence of phosphate ions. Spectroscopic studies indicated NADH:molecular oxygen reductase activity resulted in the production of the superoxide radical, detected as the formation of adrenochrome from epinephrine and by the formation of adrenochrome from epinephrine and by the reduction of nitroblue tetrazolium, both of which could be inhibited by the addition of superoxide dismutase and were unaffected by the addition of catalase. Direct observation of superoxide production using spin-trapping in combination with EPR spectroscopy resulted in the detection of the spin adduct 5.5-dimethyl-5-hydroxy-1-pyrrolidinyloxy (DMPO-OH). The formation of this spin adduct was abolished either in the absence of nitrate reductase, NADH, or DMPO or the the addition of superoxide dismutase or nitrate and was greatly reduced by the presence of cyanide. Inclusion of catalase or ethanol had no effect on the formation of the spin adduct. These results indicate that nitrate reductase can utilize molecular oxygen as an electron acceptor and that the product, O2.(-), is primarily generated via the Mopterin center.

  4. Kinetic and Structural Characterization of Dihydrofolate Reductase from Streptococcus pneumoniae

    PubMed Central

    Lee, Jeeyeon; Yennawar, Neela H.; Gam, Jongsik; Benkovic, Stephen J.

    2013-01-01

    Drug resistance associated with dihydrofolate reductase (DHFR) has emerged as a critical issue in the treatment of bacterial infections. In our efforts to understand the mechanism of a drug-resistant dihydrofolate reductase (DHFR) from a pathogenic bacterial source, we report the first kinetic characterization of Streptococcus pneumoniae DHFR (spDHFR) along with its X-ray structure. This study revealed that the kinetic properties of spDHFR were significantly different from E. coli DHFR. The product (tetrahydrofolate) dissociation step that is the rate limiting step in the E. coli DHFR is significantly accelerated in spDHFR so that hydride transfer or a preceding step is rate limiting. Comparison of the binding parameters of this enzyme to a mutant spDHFR (Sp9) confirmed that the Leu100 residue in spDHFR is the critical element for the trimethoprim (TMP) resistance. Steady-state kinetics exhibited a pH dependence in kcat, which prompted us to elucidate the role of the new catalytic residue (His33) in the active site of spDHFR. Structural data of the Sp9 mutant in complex with NADPH and methotrexate confirmed the participation of His33 in a hydrogen bonding network involving a water molecule, the hydroxyl group of Thr119, and carboxylate ion of Glu30. Sequence analysis of the DHFR superfamily revealed that the His residue is the major amino acid component at this position and is found mostly in pathogenic bacterial DHFRs. A mutation of Val100 to Leu demonstrated a steric clash of the leucine side chain with the side chains of Ile8 and Phe34, rationalizing weaker binding of trimethoprim to Leu100 DHFR. Understanding the role of specific amino acids in the active site coupled with detailed structural analysis will inform us on how to better design inhibitors targeting drug resistant pathogenic bacterial DHFRs. PMID:19950924

  5. FRUCTOSE-6-PHOSPHATE REDUCTASE FROM SALMONELLA GALLINARUM

    PubMed Central

    Zancan, Glaci T.; Bacila, Metry

    1964-01-01

    Zancan, Glaci T. (Universidade do Paraná, Curitiba, Paraná, Brazil), and Metry Bacila. Fructose-6-phosphate reductase from Salmonella gallinarum. J. Bacteriol. 87:614–618. 1964.—A fructose-6-phosphate reductase present in cell-free extracts of Salmonella gallinarum was purified approximately 42 times. The optimal pH for this enzyme is 8.0. The enzyme is specific for fructose-6-phosphate and reduced nicotinamide adenine dinucleotide (NADH). The dissociation constants are 1.78 × 10−4m for fructose-6-phosphate and 8.3 × 10−5m for NADH. The Q10, reaction order, and equilibrium constant were determined. The enzyme is sensitive to p-chloromercuribenzoic acid, but not to o-iodosobenzoic acid nor to N-ethylmaleimide. PMID:14127579

  6. Characterization of human platelet glutathione reductase.

    PubMed

    Moroff, G; Kosow, D P

    1978-12-08

    Glutathione reductase (NAD(P)h:oxidized glutathione oxidoreductase, EC 1.6.4.2) has been purified 1000-fold from the cytoplasmic fraction of human platelets. Salts, including the heretofore unreported effect of sodium citrate, activate the NADPH-dependent reduction of oxidized glutathione. Sodium citrate and monovalent salt activation appears to involve multiple sites having different binding affinities. At sub-saturating sodium phosphate, non-linear double reciprocal plots indicative of substrate activation by oxidized glutathione were observed. Initial velocity double reciprocal plots at sub-saturating and saturating concentrations of phosphate generate a family of converging lines. NADP+ is a partial inhibitor, indicating that the reduction of oxidized glutathione can proceed by more than one pathway. FMN, FAD, and riboflavin inhibit platelet glutathione reductase by influencing only the V while nitrofurantoin inhibition is associated with an increase Koxidized glutathione and a decreased V.

  7. Characterization of erythrose reductase from Yarrowia lipolytica and its influence on erythritol synthesis.

    PubMed

    Janek, Tomasz; Dobrowolski, Adam; Biegalska, Anna; Mirończuk, Aleksandra M

    2017-07-11

    Erythritol is a natural sweetener that is used in the food industry. It is produced as an osmoprotectant by bacteria and yeast. Due to its chemical properties, it does not change the insulin level in the blood, and therefore it can be safely used by diabetics. Previously, it has been shown that erythrose reductase (ER), which catalyzes the final step, plays a crucial role in erythritol synthesis. ER reduces erythrose to erythritol with NAD(P)H as a cofactor. Despite many studies on erythritol synthesis by Yarrowia lipolytica, the enzymes involved in this metabolic pathway have ever been described. The gene YALI0F18590g encoding the predicted erythrose reductase from Y. lipolytica was overexpressed, and its influence on erythritol synthesis was studied. The amino acid sequence of the Y. lipolytica ER showed a high degree of similarity to the previously described erythrose reductases from known erythritol producers, such as Candida magnoliae and Moniliella megachiliensis. Here, we found that the gene overexpression results in an enhanced titer of erythritol of 44.44 g/L (20% over the control), a yield of 0.44 g/g and productivity of 0.77 g/L/h. Moreover, on purification and characterization of the enzyme we found that it displays the highest activity at 37 °C and pH 3.0. The effects of various metal ions (Zn(2+), Cu(2+), Mn(2+), Fe(2+)) on erythrose reductase were investigated. The addition of Zn(2+) ions at 0.25 mM had a positive effect on the activity of erythrose reductase from Y. lipolytica, as well as on the erythritol production. In this study we identified, overexpressed and characterized a native erythrose reductase in Y. lipolytica. Further optimizations of this strain via metabolic pathway engineering and media optimization strategies enabled 54 g/L to be produced in a shake-flask experiment. To date, this is the first reported study employing metabolic engineering of the native gene involved in the erythritol pathway to result in a high titer of the

  8. Characterization of erythrose reductases from filamentous fungi

    PubMed Central

    2013-01-01

    Proteins with putative erythrose reductase activity have been identified in the filamentous fungi Trichoderma reesei, Aspergillus niger, and Fusarium graminearum by in silico analysis. The proteins found in T. reesei and A. niger had earlier been characterized as glycerol dehydrogenase and aldehyde reductase, respectively. Corresponding genes from all three fungi were cloned, heterologously expressed in Escherichia coli, and purified. Subsequently, they were used to establish optimal enzyme assay conditions. All three enzymes strictly require NADPH as cofactor, whereas with NADH no activity could be observed. The enzymatic characterization of the three enzymes using ten substrates revealed high substrate specificity and activity with D-erythrose and D-threose. The enzymes from T. reesei and A. niger herein showed comparable activities, whereas the one from F. graminearum reached only about a tenth of it for all tested substrates. In order to proof in vivo the proposed enzyme function, we overexpressed the erythrose reductase-encoding gene in T. reesei. An increased production of erythritol by the recombinant strain compared to the parental strain could be detected. PMID:23924507

  9. Effect of mercury, cadmium, nickel, chromium and zinc on kinetic properties of NADPH-cytochrome P450 reductase purified from leaping mullet (Liza saliens).

    PubMed

    Bozcaarmutlu, Azra; Arinç, Emel

    2007-04-01

    Information on the mechanism of metal ion inhibition of NADPH-cytochrome P450 reductase is limited. The purpose of the present paper was to elucidate in vitro effect of Hg(+2), Cd(+2), Ni(+2), Cr(+3) and Zn(+2) ions on the purified mullet NADPH-cytochrome P450 reductase. NADPH-cytochrome P450 reductase was purified from detergent-solubilized liver microsomes from leaping mullet (Liza saliens). All of the metal ions caused inhibition of the enzyme activity except Zn(+2). At 50 microM metal concentration, Hg(+2) inhibited the cytochrome P450 reductase activity completely (100%), while, at the same concentrations, Cd(+2), Cr(+3) and Ni(+2) caused 66%, 65% and 37% inhibition, respectively. At 50 microM metal concentration, Zn(+2) had no apparent effect on cytochrome P450 reductase activity. The IC(50) values of HgCl(2), CrCl(3), CdCl(2) and NiCl(2) were estimated to be 0.07 microM, 24 microM, 33 microM and 143 microM, respectively. Of the metal ions tested, Hg(+2) exhibited much higher inhibitory effect at lower concentrations, so it was evidently a more potent inhibitor than the others. All four metal ions displayed noncompetitive type of inhibition mechanism for the purified reductase as analyzed by Dixon plot. K(i) values of Hg(+2), Cr(+3), Cd(+2), and Ni(+2) were calculated from Dixon plots as 0.048 microM, 18 microM, 73 microM and 329 microM, respectively.

  10. Separation of mercury from aqueous mercuric chloride solutions by onion skins

    SciTech Connect

    Asai, S.; Konishi, Y.; Tomisaki, H.; Nakanishi, M.

    1986-01-01

    The separation of mercury from aqueous HgCl/sub 2/ solutions by onion skins (outermost coat) was studied both experimentally and theoretically. The distribution equilibria were measured by the batchwise method. The experimental results revealed that onion skin is a useful material for separating mercury from aqueous systems. The distribution data obtained at 25/sup 0/C were analyzed by using the theory based on the law of mass action. The separation of dissolved mercury by onion skins was found to be a process accompanied by an ion-exchange reaction of the cationic complex HgCl/sup +/ and an adsorption of the neutral complex HgCl/sub 2/. The equilibrium constants of the ion-exchange and adsorption processes at 25/sup 0/C and the mercury-binding capacity of onion skins were determined. Further, it was found that the distribution equilibrium of mercury is comparatively insensitive to temperature.

  11. A Ferredoxin Disulfide Reductase Delivers Electrons to the Methanosarcina barkeri Class III Ribonucleotide Reductase

    PubMed Central

    2015-01-01

    Two subtypes of class III anaerobic ribonucleotide reductases (RNRs) studied so far couple the reduction of ribonucleotides to the oxidation of formate, or the oxidation of NADPH via thioredoxin and thioredoxin reductase. Certain methanogenic archaea contain a phylogenetically distinct third subtype of class III RNR, with distinct active-site residues. Here we report the cloning and recombinant expression of the Methanosarcina barkeri class III RNR and show that the electrons required for ribonucleotide reduction can be delivered by a [4Fe-4S] protein ferredoxin disulfide reductase, and a conserved thioredoxin-like protein NrdH present in the RNR operon. The diversity of class III RNRs reflects the diversity of electron carriers used in anaerobic metabolism. PMID:26536144

  12. Methionine sulfoxide reductase contributes to meeting dietary methionine requirements

    PubMed Central

    Zhao, Hang; Kim, Geumsoo; Levine, Rodney L.

    2012-01-01

    Methionine sulfoxide reductases are present in all aerobic organisms. They contribute to antioxidant defenses by reducing methionine sulfoxide in proteins back to methionine. However, the actual in vivo roles of these reductases are not well defined. Since methionine is an essential amino acid in mammals, we hypothesized that methionine sulfoxide reductases may provide a portion of the dietary methionine requirement by recycling methionine sulfoxide. We used a classical bioassay, the growth of weanling mice fed diets varying in methionine, and applied it to mice genetically engineered to alter the levels of methionine sulfoxide reductase A or B1. Mice of all genotypes were growth retarded when raised on chow containing 0.10% methionine instead of the standard 0.45% methionine. Retardation was significantly greater in knockout mice lacking both reductases. We conclude that the methionine sulfoxide reductases can provide methionine for growth in mice with limited intake of methionine, such as may occur in the wild. PMID:22521563

  13. Technical Note: Could benzalkonium chloride be a suitable alternative to mercuric chloride for preservation of seawater samples?

    NASA Astrophysics Data System (ADS)

    Gloël, J.; Robinson, C.; Tilstone, G. H.; Tarran, G.; Kaiser, J.

    2015-08-01

    Instrumental equipment unsuitable or unavailable for fieldwork as well as lack of ship space can necessitate the preservation of seawater samples prior to analysis in a shore-based laboratory. Mercuric chloride (HgCl2) is routinely used for such preservation, but its handling and subsequent disposal incur significant risks and expense. Benzalkonium chloride (BAC) has been used previously for freshwater samples. Here, we assess BAC as a less hazardous alternative microbial inhibitor for marine samples prior to the measurement of oxygen-to-argon (O2/Ar) ratios, as used for the determination of plankton net community production. BAC at a concentration of 50 mg dm-3 inhibited microbial activity for at least three days in seawater with chlorophyll a (Chl a) concentrations up to 1 mg m-3, possibly longer when Chl a concentrations were lower. BAC concentrations of 100 and 200 mg dm-3 were no more effective than 50 mg dm-3. With fewer risks to human health and the environment, and no requirement for expensive waste disposal, BAC could be a viable alternative to HgCl2 for short-term preservation of seawater samples, but is not a replacement for HgCl2 in the case of oxygen triple isotope analysis, which requires storage over weeks to months. In any event, further tests on a case-by-case basis should be undertaken if use of BAC was considered, since its inhibitory activity may depend on concentration and composition of the microbial community.

  14. Technical note: Could benzalkonium chloride be a suitable alternative to mercuric chloride for preservation of seawater samples?

    NASA Astrophysics Data System (ADS)

    Gloël, J.; Robinson, C.; Tilstone, G. H.; Tarran, G.; Kaiser, J.

    2015-12-01

    Instrumental equipment unsuitable or unavailable for fieldwork as well as lack of ship space can necessitate the preservation of seawater samples prior to analysis in a shore-based laboratory. Mercuric chloride (HgCl2) is routinely used for such preservation, but its handling and subsequent disposal incur environmental risks and significant expense. There is therefore a strong motivation to find less hazardous alternatives. Benzalkonium chloride (BAC) has been used previously as microbial inhibitor for freshwater samples. Here, we assess the use of BAC for marine samples prior to the measurement of oxygen-to-argon (O2 / Ar) ratios, as used for the determination of biological net community production. BAC at a concentration of 50 mg dm-3 inhibited microbial activity for at least 3 days in samples tested with chlorophyll a (Chl a) concentrations up to 1 mg m-3. BAC concentrations of 100 and 200 mg dm-3 were no more effective than 50 mg dm-3. With fewer risks to human health and the environment, and no requirement for expensive waste disposal, BAC could be a viable alternative to HgCl2 for short-term preservation of seawater samples, but is not a replacement for HgCl2 in the case of oxygen triple isotope analysis, which requires storage over weeks to months. In any event, further tests on a case-by-case basis should be undertaken if use of BAC was considered, since its inhibitory activity may depend on concentration and composition of the microbial community.

  15. Performance of room temperature mercuric iodide /HgI2/ detectors in the ultralow-energy X-ray region

    NASA Technical Reports Server (NTRS)

    Dabrowski, A. J.; Barton, J. B.; Huth, G. C.; Whited, R.; Ortale, C.; Economou, T. E.; Turkevich, A. L.; Iwanczyk, J. S.

    1981-01-01

    Experiments have been done to study the performance of mercuric iodide (HgI2) detectors in the ultralow-energy X-ray region. Energy resolution values of 245 eV (FWHM) for the Mg K-alpha X-ray line at 1.25 keV and 225 eV (FWHM) for the electronic noise linewidth have been obtained for an HgI2 detector with painted carbon contacts using a pulsed-light feedback preamplifier; the whole system was operated at room temperature. The resolution values in the ultralow-energy region are still limited by electronic noise of the system. In an attempt to minimize X-ray attenuation in the front contact, detectors were prepared with thin evaporated Pd contacts. These detectors show a pronounced low-energy tailing of the photopeak below a few keV, in contrast to the spectra obtained by detectors with carbon contact. An attempt has been made to explain the tailing effect starting with models wich have been proposed to describe similar effects in Ge detectors.

  16. Th1/Th2 balance in mouse delayed-type hypersensitivity model with mercuric chloride via skin and oral mucosa.

    PubMed

    Ukichi, Kenichirou; Okamura, Taito; Fukushima, Daihei; Morimoto, Mitsuaki; Yamane, Gen-Yuki; Takahashi, Shinichi

    2011-01-01

    In order to compare delayed-type hypersensitivity (DTH) among different exposure sites, we evaluated the sensitization potency of mercuric chloride (HgCl(2)) via exposure to the skin, or oral or esophageal mucosa using the mouse ear swelling test. Furthermore, we investigated in vitro splenocyte proliferation reaction and cytokine profile in HgCl(2)-exposed and control mice. Sensitization with HgCl(2) was established via the skin and oral mucosa but not via the esophageal mucosa. The splenocyte proliferation reaction was significantly enhanced to a similar degree in skin and oral mucosa-sensitized mice compared with in the control mice. IL-10 levels from cultured splenocytes were significantly increased in skin and oral mucosa-sensitized mice compared with those in control mice, whilst IFN-γ significantly increased only in splenocytes from skin-sensitized mice. These results suggest that exposure of the skin or oral mucosa to HgCl(2) can induce DTH, but that Th1/Th2 balance differs according to the site of antigen exposure.

  17. Performance of room temperature mercuric iodide /HgI2/ detectors in the ultralow-energy X-ray region

    NASA Technical Reports Server (NTRS)

    Dabrowski, A. J.; Barton, J. B.; Huth, G. C.; Whited, R.; Ortale, C.; Economou, T. E.; Turkevich, A. L.; Iwanczyk, J. S.

    1981-01-01

    Experiments have been done to study the performance of mercuric iodide (HgI2) detectors in the ultralow-energy X-ray region. Energy resolution values of 245 eV (FWHM) for the Mg K-alpha X-ray line at 1.25 keV and 225 eV (FWHM) for the electronic noise linewidth have been obtained for an HgI2 detector with painted carbon contacts using a pulsed-light feedback preamplifier; the whole system was operated at room temperature. The resolution values in the ultralow-energy region are still limited by electronic noise of the system. In an attempt to minimize X-ray attenuation in the front contact, detectors were prepared with thin evaporated Pd contacts. These detectors show a pronounced low-energy tailing of the photopeak below a few keV, in contrast to the spectra obtained by detectors with carbon contact. An attempt has been made to explain the tailing effect starting with models wich have been proposed to describe similar effects in Ge detectors.

  18. Mercury Toxicity and Contamination of Households from the Use of Skin Creams Adulterated with Mercurous Chloride (Calomel)

    PubMed Central

    Copan, Lori; Fowles, Jeff; Barreau, Tracy; McGee, Nancy

    2015-01-01

    Inorganic mercury, in the form of mercurous chloride, or calomel, is intentionally added to some cosmetic products sold through informal channels in Mexico and the US for skin lightening and acne treatment. These products have led to multiple cases of mercury poisoning but few investigations have addressed the contamination of cream users’ homes. We report on several cases of mercury poisoning among three Mexican-American families in California from use of mercury-containing skin creams. Each case resulted in widespread household contamination and secondary contamination of family members. Urine mercury levels in cream users ranged from 37 to 482 µg/g creatinine and in non-users from non-detectable to 107 µg/g creatinine. Air concentrations of up to 8 µg/m3 of mercury within homes exceeded the USEPA/ATSDR health-based guidance and action level of <1.0 μg/m3. Mercury contamination of cream users’ homes presented a multi-pathway exposure environment to residents. Homes required extensive decontamination, including disposal of most household items, to achieve acceptable air levels. The acceptable air levels used were not designed to consider multi-pathway exposure scenarios. These findings support that the calomel is able to change valence form to elemental mercury and volatilize once exposed to the skin or surfaces in the indoor environment. PMID:26364641

  19. Measurement of the characteristic X ray of oxygen and other ultrasoft X rays using mercuric iodide detectors

    NASA Technical Reports Server (NTRS)

    Iwanczyk, J. S.; Dabrowski, A. J.; Huth, G. C.; Economou, T. E.

    1985-01-01

    This letter reports the detection and resolution of the characteristic X-ray of oxygen at 523 eV and other ultrasoft X-rays (photons energy less than 1 keV) using radiation detectors fabricated from the compound semi-insulator mercuric iodide (HgI2). These detectors are capable of operation at room ambient but in these experiments were slightly cooled using a Peltier element to 0 C. A pulsed light feedback preamplifier with a Peltier element cooled (to -30 deg) first stage field-effect transistor was used to amplify signals from the detector. Overall system noise level was 185 eV (full width at half-maximum) limited by the temperature of the first stage field-effect transistor. With optimal cooling of this element the characteristic X-ray of carbon at 282 eV should be measurable. These results would seem to be important in measurement of biological samples in electron column instruments.

  20. Mercury Toxicity and Contamination of Households from the Use of Skin Creams Adulterated with Mercurous Chloride (Calomel).

    PubMed

    Copan, Lori; Fowles, Jeff; Barreau, Tracy; McGee, Nancy

    2015-09-02

    Inorganic mercury, in the form of mercurous chloride, or calomel, is intentionally added to some cosmetic products sold through informal channels in Mexico and the US for skin lightening and acne treatment. These products have led to multiple cases of mercury poisoning but few investigations have addressed the contamination of cream users' homes. We report on several cases of mercury poisoning among three Mexican-American families in California from use of mercury-containing skin creams. Each case resulted in widespread household contamination and secondary contamination of family members. Urine mercury levels in cream users ranged from 37 to 482 µg/g creatinine and in non-users from non-detectable to 107 µg/g creatinine. Air concentrations of up to 8 µg/m³ of mercury within homes exceeded the USEPA/ATSDR health-based guidance and action level of <1.0 μg/m³. Mercury contamination of cream users' homes presented a multi-pathway exposure environment to residents. Homes required extensive decontamination, including disposal of most household items, to achieve acceptable air levels. The acceptable air levels used were not designed to consider multi-pathway exposure scenarios. These findings support that the calomel is able to change valence form to elemental mercury and volatilize once exposed to the skin or surfaces in the indoor environment.

  1. Acute effects of mercuric chloride on intracellular GSH levels and mercury distribution in the fish Oreochromic aureus

    SciTech Connect

    Allen, P.; Min, S.Y.; Keong, W.M.

    1988-02-01

    In recent years there has been much interest in the effects of trace metals on intracellular levels of reduced glutathione (GSH). Most of the research has been performed on rats. As GSH is ubiquitous in living organisms it is of interest to establish a relationship between mercury intoxication and intracellular GSH levels in fish; especially as fish living in rivers and coastal areas are often expose to mercury as an aquatic pollutant. The role of GSH in fish trace metal toxicity has not been thoroughly investigated. The distribution of total glutathione (oxidized + reduced) in selected black sea bass organs seems to follow the established pattern for mammalian organs. Thus, it would appear that teleostian and mammalian glutathione metabolism may have many similarities. There are few reports concerning the effects of mercury during the first few hours of exposure. The aim of this investigation is to establish any changes in organ GSH and mercury levels following just 2 h exposure to mercuric chloride (HgCl/sub 2/).

  2. Performance of room temperature mercuric iodide /HgI2/ detectors in the ultralow-energy X-ray region

    NASA Astrophysics Data System (ADS)

    Dabrowski, A. J.; Barton, J. B.; Huth, G. C.; Whited, R.; Ortale, C.; Economou, T. E.; Turkevich, A. L.; Iwanczyk, J. S.

    1981-02-01

    Experiments have been done to study the performance of mercuric iodide (HgI2) detectors in the ultralow-energy X-ray region. Energy resolution values of 245 eV (FWHM) for the Mg K-alpha X-ray line at 1.25 keV and 225 eV (FWHM) for the electronic noise linewidth have been obtained for an HgI2 detector with painted carbon contacts using a pulsed-light feedback preamplifier; the whole system was operated at room temperature. The resolution values in the ultralow-energy region are still limited by electronic noise of the system. In an attempt to minimize X-ray attenuation in the front contact, detectors were prepared with thin evaporated Pd contacts. These detectors show a pronounced low-energy tailing of the photopeak below a few keV, in contrast to the spectra obtained by detectors with carbon contact. An attempt has been made to explain the tailing effect starting with models wich have been proposed to describe similar effects in Ge detectors.

  3. Adsorption kinetic and equilibrium study for removal of mercuric chloride by CuCl2-impregnated activated carbon sorbent.

    PubMed

    Li, Xin; Liu, Zhouyang; Lee, Joo-Youp

    2013-05-15

    The intrinsic adsorption kinetics of mercuric chloride (HgCl2) was studied for raw, 4% and 10% CuCl2-impregnated activated carbon (CuCl2-AC) sorbents in a fixed-bed system. An HgCl2 adsorption kinetic model was developed for the AC sorbents by taking into account the adsorption kinetics, equilibrium, and internal and external mass transfer. The adsorption kinetic constants determined from the comparisons between the simulation and experimental results were 0.2, 0.3, and 0.5m(3)/(gs) for DARCO-HG, 4%(wt), and 10%(wt) CuCl2-AC sorbents, respectively, at 140 °C. CuCl2 loading was found to slightly increase the adsorption kinetic constant or at least not to decrease it. The HgCl2 equilibrium adsorption data based on the Langmuir isotherm show that high CuCl2 loading can result in high binding energy of the HgCl2 adsorption onto the carbon surface. The adsorption equilibrium constant was found to increase by ~10 times when CuCl2 loading varied from 0 to 10%(wt), which led to a decrease in the desorption kinetic constant (k2) by ~10 times and subsequently the desorption rate by ~50 times. Intraparticle pore diffusion considered in the model showed good accuracy, allowing for the determination of intrinsic HgCl2 adsorption kinetics.

  4. Study of mercuric iodide near melting using differential scanning calorimetry Raman spectroscopy, and x-ray diffraction

    SciTech Connect

    Burger, A.; Morgan, S.; Jiang, H.; Silberman, E.; Schnieber, M.; van den Berg, L.; Keller, L.; Wagner, C.N.J.

    1987-01-01

    High-temperature studies of mercuric iodide (HgI/sub 2/) involving differential scanning calorimetry (DSC), Raman spectroscopy, and x- ray powder diffraction have failed to confirm the existence of a red-colored tetragonal high-temperature phase called ..cap alpha.. '-HgI/sub 2/ reported by S.N. Toubektsis et al., (S.N. Toubektsis, E.K. Polychroniadis, and N.A. Economou, J. Appl. Phys., 58(5) (1985) 2070), using DSC measurements. The multiple DSC peaks near melting reported by Toubektsis are found by the present authors only if the sample is heated in a stainless-steel container. Using a Pyrex container or inserting a platinum foil between the HgI/sub 2/ and the stainless-steel container yields only one sharp, single DSC peak at the melting point. The nonexistence of the ..cap alpha..' phase is confirmed by high- temperature x-ray diffraction and Raman spectroscopy performed in the vicinity of the melting point which clearly indicate the existence of the yellow orthorhombic ..beta..-HgI/sub 2/ phase only. The experimental high-temperature DSC, Raman, and x-ray diffraction data are presented and discussed. 14 refs., 8 figs., 2 tabs.

  5. Measurement of the characteristic X ray of oxygen and other ultrasoft X rays using mercuric iodide detectors

    NASA Technical Reports Server (NTRS)

    Iwanczyk, J. S.; Dabrowski, A. J.; Huth, G. C.; Economou, T. E.

    1985-01-01

    This letter reports the detection and resolution of the characteristic X-ray of oxygen at 523 eV and other ultrasoft X-rays (photons energy less than 1 keV) using radiation detectors fabricated from the compound semi-insulator mercuric iodide (HgI2). These detectors are capable of operation at room ambient but in these experiments were slightly cooled using a Peltier element to 0 C. A pulsed light feedback preamplifier with a Peltier element cooled (to -30 deg) first stage field-effect transistor was used to amplify signals from the detector. Overall system noise level was 185 eV (full width at half-maximum) limited by the temperature of the first stage field-effect transistor. With optimal cooling of this element the characteristic X-ray of carbon at 282 eV should be measurable. These results would seem to be important in measurement of biological samples in electron column instruments.

  6. Structure of octaheme cytochrome c nitrite reductase from Thioalkalivibrio nitratireducens in a complex with phosphate

    SciTech Connect

    Trofimov, A. A.; Polyakov, K. M.; Boiko, K. M.; Filimonenkov, A. A.; Dorovatovskii, P. V.; Tikhonova, T. V.; Popov, V. O.; Koval'chuk, M. V.

    2010-01-15

    Octaheme cytochrome c nitrite reductase from Thioalkalivibrio nitratireducens (TvNiR) catalyzes the reduction of nitrite and hydroxylamine to ammonia. The structures of the free enzyme and of the enzyme in complexes with the substrate (nitrite ion) and the inhibitor (azide ion) have been solved previously. In this study we report the structures of the oxidized complex of TvNiR with phosphate and of this complex reduced by europium(II) chloride (1.8- and 2.0-A resolution, the R factors are 15.9 and 16.7%, respectively) and the structure of the enzyme in the complex with cyanide (1.76-A resolution, the R factor is 16.5%), which was prepared by soaking a crystal of the oxidized phosphate complex of TvNiR. In the active site of the enzyme, the phosphate ion binds to the iron ion of the catalytic heme and to the side chains of the catalytic residues Arg131, Tyr303, and His361. The cyanide ion is coordinated to the heme-iron ion and is hydrogen bonded to the residue His361. In the structure of reduced TvNiR, the phosphate ion is bound in the same manner as in the structure of oxidized TvNiR, and the nine{sub c}oordinated europium ion is located on the surface of one of the crystallographically independent monomers of the enzyme.

  7. Structural Elucidation of Chalcone Reductase and Implications for Deoxychalcone Biosynthesis

    PubMed Central

    Bomati, Erin K.; Austin, Michael B.; Bowman, Marianne E.; Dixon, Richard A.; Noel, Joseph P.

    2010-01-01

    4,2′,4′,6′-tetrahydroxychalcone (chalcone) and 4,2′,4′-trihydroxychalcone (deoxychalcone) serve as precursors of ecologically important flavonoids and isoflavonoids. Deoxychalcone formation depends on chalcone synthase and chalcone reductase; however, the identity of the chalcone reductase substrate out of the possible substrates formed during the multistep reaction catalyzed by chalcone synthase remains experimentally elusive. We report here the three-dimensional structure of alfalfa chalcone reductase bound to the NADP+ cofactor and propose the identity and binding mode of its substrate, namely the non-aromatized coumaryl-trione intermediate of the chalcone synthase-catalyzed cyclization of the fully extended coumaryl-tetraketide thioester intermediate. In the absence of a ternary complex, the quality of the refined NADP+-bound chalcone reductase structure serves as a template for computer-assisted docking to evaluate the likelihood of possible substrates. Interestingly, chalcone reductase adopts the three-dimensional structure of the aldo/keto reductase superfamily. The aldo/keto reductase fold is structurally distinct from all known ketoreductases of fatty acid biosynthesis, which instead belong to the short-chain dehydrogenase/reductase superfamily. The results presented here provide structural support for convergent functional evolution of these two ketoreductases that share similar roles in the biosynthesis of fatty acids/polyketides. In addition, the chalcone reductase structure represents the first protein structure of a member of the aldo/ketoreductase 4 family. Therefore, the chalcone reductase structure serves as a template for the homology modeling of other aldo/ketoreductase 4 family members, including the reductase involved in morphine biosynthesis, namely codeinone reductase. PMID:15970585

  8. Limited proteolysis of the nitrate reductase from spinach leaves.

    PubMed

    Kubo, Y; Ogura, N; Nakagawa, H

    1988-12-25

    The functional structure of assimilatory NADH-nitrate reductase from spinach leaves was studied by limited proteolysis experiments. After incubation of purified nitrate reductase with trypsin, two stable products of 59 and 45 kDa were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The fragment of 45 kDa was purified by Blue Sepharose chromatography. NADH-ferricyanide reductase and NADH-cytochrome c reductase activities were associated with this 45-kDa fragment which contains FAD, heme, and NADH binding fragment. After incubation of purified nitrate reductase with Staphylococcus aureus V8 protease, two major peaks were observed by high performance liquid chromatography size exclusion gel filtration. FMNH2-nitrate reductase and reduced methyl viologen-nitrate reductase activities were associated with the first peak of 170 kDa which consists of two noncovalently associated (75-90-kDa) fragments. NADH-ferricyanide reductase activity, however, was associated with the second peak which consisted of FAD and NADH binding sites. Incubation of the 45-kDa fragment with S. aureus V8 protease produced two major fragments of 28 and 14 kDa which contained FAD and heme, respectively. These results indicate that the molybdenum, heme, and FAD components of spinach nitrate reductase are contained in distinct domains which are covalently linked by exposed hinge regions. The molybdenum domain appears to be important in the maintenance of subunit interactions in the enzyme complex.

  9. Mercuric iodide (HgI/sub 2/) semiconductor devices as charged-particle detectors

    SciTech Connect

    Becchetti, F.D.; Raymond, R.S.; Ristinen, R.A.; Schnepple, W.F.; Ortale, C.

    1981-01-01

    The properties of HgI/sub 2/ semiconductor devices as charged particle detectors have been investigated. Nearly linear energy response with FWHM resolution of 5 to 15% is observed for /sup 1/ /sup 2/H and /sup 3/ /sup 4/He ions, E < 40 MeV. Fast proton damage is observed for > 10/sup 10/ protons/cm/sup 2/. However, based on measurements with two HgI/sub 2/ detectors, little fast neutron damage is apparent at fluences up to 10/sup 15/ neutrons/cm/sup 2/. This suggests considerably greater resistance to radiation damage than is observed for Si and other solid state devices.

  10. Structure and function of NADPH-cytochrome P450 reductase and nitric oxide synthase reductase domain

    SciTech Connect

    Iyanagi, Takashi . E-mail: iyanagi@spring8.or.jp

    2005-12-09

    NADPH-cytochrome P450 reductase (CPR) and the nitric oxide synthase (NOS) reductase domains are members of the FAD-FMN family of proteins. The FAD accepts two reducing equivalents from NADPH (dehydrogenase flavin) and FMN acts as a one-electron carrier (flavodoxin-type flavin) for the transfer from NADPH to the heme protein, in which the FMNH {sup {center_dot}}/FMNH{sub 2} couple donates electrons to cytochrome P450 at constant oxidation-reduction potential. Although the interflavin electron transfer between FAD and FMN is not strictly regulated in CPR, electron transfer is activated in neuronal NOS reductase domain upon binding calmodulin (CaM), in which the CaM-bound activated form can function by a similar mechanism to that of CPR. The oxygenated form and spin state of substrate-bound cytochrome P450 in perfused rat liver are also discussed in terms of stepwise one-electron transfer from CPR. This review provides a historical perspective of the microsomal mixed-function oxidases including CPR and P450. In addition, a new model for the redox-linked conformational changes during the catalytic cycle for both CPR and NOS reductase domain is also discussed.

  11. Molecular dissection of a putative iron reductase from Desulfotomaculum reducens MI-1

    PubMed Central

    Li, Zhi; Kim, David D.; Nelson, Ornella D.; Otwell, Anne E.; Richardson, Ruth E.; Callister, Stephen J.; Lin, Hening

    2015-01-01

    Desulfotomaculum reducens MI-1 is a Firmicute strain capable of reducing a variety of heavy metal ions and has a great potential in heavy metal bioremediation. We recently identified Dred_2421 as a potential iron reductase through proteomic study of D. reducens. The current study examines its iron-reduction mechanism. Dred_2421, like its close homolog from Escherichia coli (2, 4-dienoyl-CoA reductase), has an FMN-binding N-terminal domain (NTD), an FAD-binding C-terminal domain (CTD), and a 4Fe-4S cluster between the two domains. To understand the mechanism of the iron-reduction activity and the role of each domain, we generated a series of variants for each domain and investigated their iron-reduction activity. Our results suggest that CTD is the main contributor of the iron-reduction activity, and that NTD and the 4Fe-4S cluster are not directly involved in such activity. This study provides a mechanistic understanding of the iron-reductase activity of Dred_2421 and may also help to elucidate other physiological activities this enzyme may have. PMID:26454174

  12. Molecular dissection of a putative iron reductase from Desulfotomaculum reducens MI-1.

    PubMed

    Li, Zhi; Kim, David D; Nelson, Ornella D; Otwell, Anne E; Richardson, Ruth E; Callister, Stephen J; Lin, Hening

    2015-11-20

    Desulfotomaculum reducens MI-1 is a Firmicute strain capable of reducing a variety of heavy metal ions and has a great potential in heavy metal bioremediation. We recently identified Dred_2421 as a potential iron reductase through proteomic study of D. reducens. The current study examines its iron-reduction mechanism. Dred_2421, like its close homolog from Escherichia coli (2, 4-dienoyl-CoA reductase), has an FMN-binding N-terminal domain (NTD), an FAD-binding C-terminal domain (CTD), and a 4Fe-4S cluster between the two domains. To understand the mechanism of the iron-reduction activity and the role of each domain, we generated a series of variants for each domain and investigated their iron-reduction activity. Our results suggest that CTD is the main contributor of the iron-reduction activity, and that NTD and the 4Fe-4S cluster are not directly involved in such activity. This study provides a mechanistic understanding of the iron-reductase activity of Dred_2421 and may also help to elucidate other physiological activities this enzyme may have.

  13. Evolution of the Ferric Reductase Domain (FRD) Superfamily: Modularity, Functional Diversification, and Signature Motifs

    PubMed Central

    Zhang, Xuezhi; Krause, Karl-Heinz; Xenarios, Ioannis; Soldati, Thierry; Boeckmann, Brigitte

    2013-01-01

    A heme-containing transmembrane ferric reductase domain (FRD) is found in bacterial and eukaryotic protein families, including ferric reductases (FRE), and NADPH oxidases (NOX). The aim of this study was to understand the phylogeny of the FRD superfamily. Bacteria contain FRD proteins consisting only of the ferric reductase domain, such as YedZ and short bFRE proteins. Full length FRE and NOX enzymes are mostly found in eukaryotic cells and all possess a dehydrogenase domain, allowing them to catalyze electron transfer from cytosolic NADPH to extracellular metal ions (FRE) or oxygen (NOX). Metazoa possess YedZ-related STEAP proteins, possibly derived from bacteria through horizontal gene transfer. Phylogenetic analyses suggests that FRE enzymes appeared early in evolution, followed by a transition towards EF-hand containing NOX enzymes (NOX5- and DUOX-like). An ancestral gene of the NOX(1-4) family probably lost the EF-hands and new regulatory mechanisms of increasing complexity evolved in this clade. Two signature motifs were identified: NOX enzymes are distinguished from FRE enzymes through a four amino acid motif spanning from transmembrane domain 3 (TM3) to TM4, and YedZ/STEAP proteins are identified by the replacement of the first canonical heme-spanning histidine by a highly conserved arginine. The FRD superfamily most likely originated in bacteria. PMID:23505460

  14. NADPH-Cytochrome P450 Oxidoreductase: Prototypic Member of the Diflavin Reductase Family

    PubMed Central

    Iyanagi, Takashi; Xia, Chuanwu; Kim, Jung-Ja P.

    2012-01-01

    NADPH-cytochrome P450 oxidoreductase (CYPOR) and nitric oxide synthase (NOS), two members of the diflavin oxidoreductase family, are multi-domain enzymes containing distinct FAD and FMN domains connected by a flexible hinge. FAD accepts a hydride ion from NADPH, and reduced FAD donates electrons to FMN, which in turn transfers electrons to the heme center of cytochrome P450 or NOS oxygenase domain. Structural analysis of CYPOR, the prototype of this enzyme family, has revealed the exact nature of the domain arrangement and the role of residues involved in cofactor binding. Recent structural and biophysical studies of CYPOR have shown that the two flavin domains undergo large domain movements during catalysis. NOS isoforms contain additional regulatory elements within the reductase domain that control electron transfer through Ca2+-dependent calmodulin (CaM) binding. The recent crystal structure of an iNOS Ca2+/CaM-FMN construct, containing the FMN domain in complex with Ca2+/CaM, provided structural information on the linkage between the reductase and oxgenase domains of NOS, making it possible to model the holo iNOS structure. This review summarizes recent advances in our understanding of the dynamics of domain movements during CYPOR catalysis and the role of the NOS diflavin reductase domain in the regulation of NOS isozyme activities. PMID:22982532

  15. Structural study of the X-ray-induced enzymatic reaction of octahaem cytochrome C nitrite reductase.

    PubMed

    Trofimov, A A; Polyakov, K M; Lazarenko, V A; Popov, A N; Tikhonova, T V; Tikhonov, A V; Popov, V O

    2015-05-01

    Octahaem cytochrome c nitrite reductase from the bacterium Thioalkalivibrio nitratireducens catalyzes the reduction of nitrite to ammonium and of sulfite to sulfide. The reducing properties of X-ray radiation and the high quality of the enzyme crystals allow study of the catalytic reaction of cytochrome c nitrite reductase directly in a crystal of the enzyme, with the reaction being induced by X-rays. Series of diffraction data sets with increasing absorbed dose were collected from crystals of the free form of the enzyme and its complexes with nitrite and sulfite. The corresponding structures revealed gradual changes associated with the reduction of the catalytic haems by X-rays. In the case of the nitrite complex the conversion of the nitrite ions bound in the active sites to NO species was observed, which is the beginning of the catalytic reaction. For the free form, an increase in the distance between the oxygen ligand bound to the catalytic haem and the iron ion of the haem took place. In the case of the sulfite complex no enzymatic reaction was detected, but there were changes in the arrangement of the active-site water molecules that were presumably associated with a change in the protonation state of the sulfite ions.

  16. Ion-Ion Neutralization.

    DTIC Science & Technology

    1982-05-31

    Accession No. 3. Recipient’s Catalog Number FGL -TR-82 -0202 b- /- 4. Title (and Subtitle) 5. Type of Report & Period Covered ION-ION NEUTRALIZATION Final...few years under the terms of the grant has been the detailed study of binary ion-ion neutralization reactions involving ions of atmospheric...2TT, England. 1. INTRODUCTION Binary positive-ion negative-ion mutual neutralization viz: A+ + B->C + D (1) can be an important loss process for

  17. An improved method for the synthesis of mercurated dUTP. Enzymic synthesis of Hg-labelled DNA of high molecular weight suitable for use in an image based DNA sequencing strategy.

    PubMed

    Bridgman, A J; Petersen, G B

    1996-01-01

    The development of high-resolution scanning-probe microscopes has reawakened interest in the possibility of sequencing large nucleic acid molecules by direct imaging. Such an approach would be facilitated by the availability of effective methods for increasing contrast by labelling specific nucleotides, and the utility of introducing mercury atoms into complete DNA molecules through the enzymic polymerisation of mercurated pyrimidine deoxynucleoside triphosphates has been re-investigated. A simplified and improved method for the synthesis of a heat- and thiol-stable, mercurated derivative of deoxyuridine triphosphate in high yield and the incorporation of this precursor into full-length copies of a single-stranded phage M13 template are described. The DNA product has been fully characterised and the quantitative and specific replacement of thymidylic acid residues by the mercurated analogue demonstrated.

  18. Geochemical Results of Lysimeter Sampling at the Manning Canyon Repository in the Mercur Mining District, Utah

    USGS Publications Warehouse

    Earle, John; Choate, LaDonna

    2010-01-01

    This report presents chemical characteristics of transient unsaturated-zone water collected by lysimeter from the Manning Canyon repository site in Utah. Data collected by U.S. Geological Survey and U.S. Department of the Interior, Bureau of Land Management scientists under an intragovernmental order comprise the existing body of hydrochemical information on unsaturated-zone conditions at the site and represent the first effort to characterize the chemistry of the soil pore water surrounding the repository. Analyzed samples showed elevated levels of arsenic, barium, chromium, and strontium, which are typical of acidic mine drainage. The range of major-ion concentrations generally showed expected soil values. Although subsequent sampling is necessary to determine long-term effects of the repository, current results provide initial data concerning reactive processes of precipitation on the mine tailings and waste rock stored at the site and provide information on the effectiveness of reclamation operations at the Manning Canyon repository.

  19. Vibrational structure of dihydrofolate bound to R67 dihydrofolate reductase.

    PubMed

    Deng, H; Callender, R; Howell, E

    2001-12-28

    R67 is a Type II dihydrofolate reductase (DHFR) that catalyzes the reduction of dihydrofolate (DHF) to tetrahydrofolate by facilitating the addition of a proton to N5 of DHF and the transfer of a hydride ion from NADPH to C6. Because this enzyme is a plasmid-encoded DHFR from trimethoprim-resistant bacteria, extensive studies on R67 with various methods have been performed to elucidate its reaction mechanism. Here, Raman difference measurements, conducted on the ternary complex of R67.NADP(+).DHF believed to be an accurate mimic of the productive DHFR.NADPH.DHF complex, show that the pK(a) of N5 in the complex is less than 4. This is in clear contrast to the behavior observed in Escherichia coli DHFR, a substantially more efficient enzyme, where the pK(a) of bound DHF at N5 is increased to 6.5 compared with its solution value of 2.6. A comparison of the ternary complexes in R67 and E. coli DHFRs suggests that enzymic raising of the pK(a) at N5 can significantly increase the catalytic efficiency of the hydride transfer step. However, R67 shows that even without such a strategy an effective DHFR can still be designed.

  20. Nitrate Reductase of Primary Roots of Red Spruce Seedlings 1

    PubMed Central

    Yandow, Tim S.; Klein, Richard M.

    1986-01-01

    Nitrate reductase activity (NRA) was found in primary roots, but not in foliage of red spruce (Picea rubens Sarg.) seedlings. Nitrate induced NRA:NH4+ did not induce and slightly depressed NRA in older seedlings. Induction required 8 hours and, once induced, NRA decreased slowly in the absence of exogenous NO3−. Seedlings were grown in perlite with a complete nutrient solution containing NH4+ to limit NR induction. Established seedlings were stressed with nutrient solutions at pH 3, 4, or 5 supplemented with Cl− salts of Al, Cd, Pb, or Zn each at two concentrations. NRA in primary root tips was measured at 2, 14, 28, and 42 days. NRA induction was greatest at pH 3, and remained high during the period of study. NRA induction at pH 4 was lower. Metal ions suppressed NRA at pH 3 and 5, but enhanced NRA at pH 4. It is concluded that acidity and soluble metals in the root environment of red spruce are unlikely to be important factors in nitrogen transformations in red spruce roots. PMID:16664891

  1. Mechanism of inhibition of purified leaping mullet (Liza saliens) NADPH-cytochrome P450 reductase by toxic metals: aluminum and thallium.

    PubMed

    Bozcaarmutlu, Azra

    2007-01-01

    Aluminum and thallium may reach life-threatening levels in aquatic systems in the near future because of their extensive use in various industrial fields. It is therefore important to study the mechanism of toxicity of aluminum and thallium on fish enzymes. To this aim, the effects of aluminum and thallium on the activity of purified leaping mullet (Liza saliens) cytochrome P450 reductase, an essential component of the important cytochrome P450 system, have been studied. Results indicated that both metal ions strongly inhibited the NADPH-cytochrome P450 reductase. The IC50 values of AlCl3 and TlCl3 were estimated to be 34 microM and 3 microM, respectively. The Lineweaver-Burk plot and Dixon plot revealed that both metal ions noncompetitively inhibited the purified mullet cytochrome P450 reductase. The K(i) values of Al3+ and Tl3+ were calculated from Dixon plots as 8.9 and 5.6 microM, respectively. The inhibitory effects of Al3+ and Tl3+ on purified cytochrome P450 reductase were partially recovered by 1 mM EDTA. Additionally, tin and magnesium were shown to have no apparent effect on purified mullet cytochrome P450 reductase.

  2. 3-Methyleneoxindole Reductase of Peas 1

    PubMed Central

    Moyed, H. S.; Williamson, Valerie

    1967-01-01

    A 100-fold purification of a reduced triphosphopyridine nucleotide/3-methyleneoxindole reductase of peas has been achieved using conventional protein fractionation procedures. Reduced diphosphopyridine nucleotide is 25-fold less effective than reduced triphosphopyridine nucleotide as the reductant. The preparation is free of other reductase activities including those linking the oxidation of reduced pyridine nucleotide coenzymes to the reduction of cytochrome c; vitamins K1, K2, and K3; O2; nitrate; oxidized glutathione; and thiazolyl blue tetrazolium. The affinity of the enzyme for 3-methyleneoxindole (Ks = 0.5 mm 3-methyleneoxindole) is relatively high. It is, therefore, reasonable to assume that 3-methyleneoxindole is the normal substrate. The enzyme is inhibited by indole-3-acetic acid, indole-3-aldehyde, and by l-naph-thaleneacetic acid. While these are not especially powerful inhibitors (K1 = 1.9-4.0 mm) the competitive relationship with 3-methyleneoxindole indicates that significant inhibition might occur at low intracellular concentrations of the substrate. PMID:6042360

  3. Enzyme toolbox: novel enantiocomplementary imine reductases.

    PubMed

    Scheller, Philipp N; Fademrecht, Silvia; Hofelzer, Sebastian; Pleiss, Jürgen; Leipold, Friedemann; Turner, Nicholas J; Nestl, Bettina M; Hauer, Bernhard

    2014-10-13

    Reducing reactions are among the most useful transformations for the generation of chiral compounds in the fine-chemical industry. Because of their exquisite selectivities, enzymatic approaches have emerged as the method of choice for the reduction of C=O and activated C=C bonds. However, stereoselective enzymatic reduction of C=N bonds is still in its infancy-it was only recently described after the discovery of enzymes capable of imine reduction. In our work, we increased the spectrum of imine-reducing enzymes by database analysis. By combining the currently available knowledge about the function of imine reductases with the experimentally uncharacterized diversity stored in protein sequence databases, three novel imine reductases with complementary enantiopreference were identified along with amino acids important for catalysis. Furthermore, their reducing capability was demonstrated by the reduction of the pharmaceutically relevant prochiral imine 2-methylpyrroline. These novel enzymes exhibited comparable to higher catalytic efficiencies than previously described enzymes, and their biosynthetic potential is highlighted by the full conversion of 2-methylpyrroline in whole cells with excellent selectivities.

  4. Functional and Phylogenetic Divergence of Fungal Adenylate-Forming Reductases

    PubMed Central

    Kalb, Daniel; Lackner, Gerald

    2014-01-01

    A key step in fungal l-lysine biosynthesis is catalyzed by adenylate-forming l-α-aminoadipic acid reductases, organized in domains for adenylation, thiolation, and the reduction step. However, the genomes of numerous ascomycetes and basidiomycetes contain an unexpectedly large number of additional genes encoding similar but functionally distinct enzymes. Here, we describe the functional in vitro characterization of four reductases which were heterologously produced in Escherichia coli. The Ceriporiopsis subvermispora serine reductase Nps1 features a terminal ferredoxin-NADP+ reductase (FNR) domain and thus belongs to a hitherto undescribed class of fungal multidomain enzymes. The second major class is characterized by the canonical terminal short-chain dehydrogenase/reductase domain and represented by Ceriporiopsis subvermispora Nps3 as the first biochemically characterized l-α-aminoadipic acid reductase of basidiomycete origin. Aspergillus flavus l-tyrosine reductases LnaA and LnbA are members of a distinct phylogenetic clade. Phylogenetic analysis supports the view that fungal adenylate-forming reductases are more diverse than previously recognized and belong to four distinct classes. PMID:25085485

  5. Structure of an integral membrane sterol reductase from Methylomicrobium alcaliphilum

    PubMed Central

    Li, Xiaochun; Roberti, Rita; Blobel, Günter

    2014-01-01

    Sterols are essential biological molecules in the majority of life forms. Sterol reductases1 including Delta-14 sterol reductase (C14SR), 7-dehydrocholesterol reductase (DHCR7) and 24-dehydrocholesterol reductase (DHCR24) reduce specific carbon-carbon double bonds of the sterol moiety using a reducing cofactor during sterol biosynthesis. Lamin B Receptor2 (LBR), an integral inner nuclear membrane protein, also contains a functional C14SR domain. Here we report the crystal structure of a Delta-14 sterol reductase (maSR1) from the methanotrophic bacterium Methylomicrobium alcaliphilum 20Z, a homolog of human C14SR, LBR, and DHCR7, with the cofactor NADPH. The enzyme contains 10 transmembrane segments (TM). Its catalytic domain comprises the C-terminal half (containing TM6-10) and envelops two interconnected pockets, one of which faces the cytoplasm and houses NADPH, while the other one is accessible from the lipid bilayer. Comparison with a soluble steroid 5β-reductase structure3 suggests that the reducing end of NADPH meets the sterol substrate at the juncture of the two pockets. A sterol reductase activity assay proves maSR1 can reduce the double bond of a cholesterol biosynthetic intermediate demonstrating functional conservation to human C14SR. Therefore, our structure as a prototype of integral membrane sterol reductases provides molecular insight into mutations in DHCR7 and LBR for inborn human diseases. PMID:25307054

  6. Structural and biochemical characterization of cinnamoyl-coa reductases

    USDA-ARS?s Scientific Manuscript database

    Cinnamoyl-coenzyme A reductase (CCR) catalyzes the reduction of hydroxycinnamoyl-coenzyme A (CoA) esters using NADPH to produce hydroxycinnamyl aldehyde precursors in lignin synthesis. The catalytic mechanism and substrate specificity of cinnamoyl-CoA reductases from sorghum (Sorghum bicolor), a str...

  7. A study of low-noise preamplifier systems for use with room temperature mercuric iodide /HgI2/ X-ray detectors

    NASA Technical Reports Server (NTRS)

    Iwanczyk, J. S.; Huth, G. C.; Del Duca, A.; Schnepple, W.; Dabrowski, A. J.

    1981-01-01

    An analysis of different preamplification systems for use with room temperature mercuric iodide X-ray detectors has been performed. Resistor-, drain-, and light-feedback preamplifiers have been studied. Energy resolution values of 295 eV (FWHM) for an Fe-55 source (5.9 keV) and 225 eV (FWHM) for a pulser have been obtained with both the detector and the input FET at room temperature using a pulsed-light feedback preamplifier. Improvement in energy resolution by cooling the input FET using a small Peltier element has been discussed.

  8. Evaluation of intestinal protozoan morphology in polyvinyl alcohol preservative: comparison of zinc sulfate- and mercuric chloride-based compounds for use in Schaudinn's fixative.

    PubMed Central

    Garcia, L S; Shimizu, R Y; Shum, A; Bruckner, D A

    1993-01-01

    As a result of disposal problems related to the use of mercury compounds, many laboratories have considered switching from mercuric chloride-based Schaudinn's and polyvinyl alcohol (PVA) stool preservatives to other non-mercury-based preservatives. The primary use for PVA-preserved specimens is the permanent stained smear, the most important technique in the routine ova and parasite examination for the identification and confirmation of intestinal protozoa. A comparison of organism recovery and morphology of the intestinal protozoa was undertaken with PVA containing either a zinc sulfate base or the "gold standard" mercuric chloride base. Paired positive fecal specimens (106 from 64 patients) were collected and examined microscopically by the trichrome stain technique. There were 161 instances in which organism trophozoite and/or cyst stages were identified and 3 in which human cells were identified. Morphology, clarity of nuclear and cytoplasmic detail, overall color differences, and the ease or difficulty in detecting intestinal protozoa in fecal debris, as well as the number of patients with a missed diagnosis, were assessed from the permanent stained smear. Overall organism morphology of the intestinal protozoa preserved in zinc sulfate-PVA was not always equal in nuclear and cytoplasmic detail or range of color after permanent staining to that seen with mercuric chloride-PVA. However, the same organisms were usually identified in both specimens, with the exception of situations in which organism numbers were characterized as rare (no organisms per 10 oil immersion fields at x1,000 magnification but at least one organism in the smear) [9 of 161 (5.6%)] or the organism was missed because of poor morphologic detail [12 of 161 (7.5%)]. In only six of these cases [6 of 161 (3.7%)] did the results involve pathogens. The patient diagnosis was missed in four cases of amebiasis and two cases of giardiasis; in both situations the organism numbers were rare. There were

  9. A study of low-noise preamplifier systems for use with room temperature mercuric iodide /HgI2/ X-ray detectors

    NASA Astrophysics Data System (ADS)

    Iwanczyk, J. S.; Huth, G. C.; del Duca, A.; Schnepple, W.; Dabrowski, A. J.

    1981-02-01

    An analysis of different preamplification systems for use with room temperature mercuric iodide X-ray detectors has been performed. Resistor-, drain-, and light-feedback preamplifiers have been studied. Energy resolution values of 295 eV (FWHM) for an Fe-55 source (5.9 keV) and 225 eV (FWHM) for a pulser have been obtained with both the detector and the input FET at room temperature using a pulsed-light feedback preamplifier. Improvement in energy resolution by cooling the input FET using a small Peltier element has been discussed.

  10. One-step green synthesis and characterization of plant protein-coated mercuric oxide (HgO) nanoparticles: antimicrobial studies

    NASA Astrophysics Data System (ADS)

    Das, Amlan Kumar; Marwal, Avinash; Sain, Divya; Pareek, Vikram

    2015-03-01

    The present study demonstrates the bioreductive green synthesis of nanosized HgO using flower extracts of an ornamental plant Callistemon viminalis. The flower extracts of Callistemon viminalis seem to be environmentally friendly, so this protocol could be used for rapid production of HgO. Till date, there is no report of synthesis of nanoparticles using flower extract of Callistemon viminalis. Mercuric acetate was taken as the metal precursor in the present experiment. The flower extract was found to act as a reducing as well as a stabilizing agent. The phytochemicals present in the flower extract act as reducing agent which include proteins, saponins, phenolic compounds, phytosterols, and flavonoids. FT-IR spectroscopy confirmed that the extract had the ability to act as a reducing agent and stabilizer for HgO nanoparticles. The formation of the plant protein-coated HgO nanoparticles was first monitored using UV-Vis absorption spectroscopy. The UV-Vis spectroscopy revealed the formation of HgO nanoparticles by exhibiting the typical surface plasmon absorption maxima at 243 nm. The average particle size formed ranges from 2 to 4 nm. The dried form of synthesized nanoparticles was further characterized using TGA, XRD, TEM, and FTIR spectroscopy. FT-IR spectra of synthesized HgO nanoparticles were performed to identify the possible bio-molecules responsible for capping and stabilization of nanoparticles, which confirm the formation of plant protein-coated HgO nanoparticles that is further corroborated by TGA study. The optical band gap of HgO nanoparticle was measured to be 2.48 eV using cutoff wavelength which indicates that HgO nanoparticles can be used in metal oxide semiconductor-based photovoltaic cells. A possible core-shell structure of the HgO nanobiocomposite has been proposed.

  11. Effect of mercuric chloride on fertilization and larval development in the River Frog, Rana heckscheri (Wright) (Anura: Ranidae)

    SciTech Connect

    Punzo, F. )

    1993-10-01

    Previous investigations have indicated that heavy metals such as copper, cadmium, lead and mercury can act as systemic toxicants in many species of wildlife. Although numerous studies have emphasized the effects of metals and pesticides on metabolism, growth, survivorship, neural processes and reproduction in a number of taxa, little information is available on the effects of sublethal concentrations of metals on the reproductive physiology of amphibians. Industrial processes and mining activities can release substantial concentrations of heavy metals such as mercury into aquatic habitats. Since most amphibians have obligate aquatic larval stages, they are exposed to pollutants discharged into the aquatic environment. Amphibians can act as accumulators of heavy metals and their larval stages are useful indicators of pollution levels in the field. What little data are available, indicate that metals can significantly reduce viability in amphibians through their actions on metabolism, development and gametogenesis. The recent concerns over worldwide declines in amphibian populations and the susceptibility of amphibian populations to environmental toxicants, led me to assess the effect of mercuric chloride, one of the most common and persistent toxicants in aquatic environments, on fertilization and larval development in the river frog, Rana heckscheri (Wright). Although there is some information on fish, very little data are available on the effects of mercury on fertilization in amphibians generally, and no published data exist for R. heckscheri. This species is a conspicuous component of the aquatic fauna of parts of the southeastern United States where mercury levels have increased significantly over the last two decades. 22 refs., 2 tabs.

  12. Exploration of strategies for implementation of screen-printed mercuric iodide converters in direct detection AMFPIs for digital breast tomosynthesis

    NASA Astrophysics Data System (ADS)

    Antonuk, Larry E.; El-Mohri, Youcef; Zhao, Qihua; Jiang, Hao

    2017-03-01

    Digital breast tomosynthesis (DBT) has become an increasingly important tool in the diagnosis of breast disease. For those DBT imaging systems based on active matrix, flat-panel imager (AMFPI) arrays, the incident radiation is detected directly or indirectly by means of an a-Se or CsI:Tl x-ray converter, respectively. While all AMFPI DBT devices provide clinically useful volumetric information, their performance is limited by the relatively modest average signal generated per interacting X ray by present converters compared to the electronic additive noise of the system. To address this constraint, we are pursuing the development of a screen-printed form of mercuric iodide (SP HgI2) which has demonstrated considerably higher sensitivities (i.e., larger average signal per interacting X ray) than those of conventional a-Se and CsI:Tl converters, as well as impressive DQE and MTF performance under mammographic irradiation conditions. A converter offering such enhanced sensitivity would greatly improve signal-to-noise performance and facilitate quantum-limited imaging down to significantly lower exposures than present AMFPI DBT systems. However, before this novel converter material can be implemented practically, challenges associated with SP HgI2 must be addressed. Most significantly, high levels of charge trapping (which lead to image lag as well as fall-off in DQE at higher exposures) need to be reduced - while improving the uniformity in pixel-to-pixel signal response as well as maintaining low dark current and otherwise favorable DQE performance. In this paper, a pair of novel strategies for overcoming the challenge of charge trapping in SP HgI2 converters are described, and initial results from empirical and calculational studies of these strategies are reported.

  13. Effects of water hardness and temperature on the acute toxicity of mercuric chloride on rainbow trout (Oncorhynchus mykiss).

    PubMed

    Terzi, Ertugrul; Verep, Bulent

    2012-07-01

    In this study, the toxicity of mercuric chloride (HgCl(2)), an important pollutant threatening water resources for many years, and the effects of water temperature and hardness on the toxicity in cultured rainbow trout Oncorhynchus mykiss (4.79 ± 0.16 g; 7.38 ± 0.24 cm; mean ± SD) were investigated at different temperatures (12 and 17°C) and hardness concentrations (35, 70 and 120 mg l(-1) as calcium carbonate, CaCO(3)). For this purpose, the acute toxicity tests were performed by 96-h static tests in different water temperatures and water hardness concentrations. For acute toxicity tests, solutions ranging from 0.4 to 1.2 mg l(-1) were used at 12°C and solutions ranging from 0.4 to 1.0 mg l(-1) at 17°C. The LC(50) values of HgCl(2) that killed 50% of rainbow trout within 96 h in the hardness concentrations of 35, 70 and 120 mg l(-1) CaCO(3) were calculated using probit analysis, and were found to be 0.725, 0.788, 0.855 mg l(-1) at 12°C and 0.670, 0.741, 0.787 mg l(-1) at 17°C, respectively. Consequently, the toxicity of HgCl(2) on rainbow trout decreased when the temperature decreased from 17 to 12°C. Toxicity increased when the hardness decreased from 120 to 35 mg l(-1) CaCO(3). In contrast to temperature, water hardness presents a negative effect on the toxicity of HgCl(2).

  14. Effects of mercuric chloride on chemiluminescent response of phagocytes and tissue lysozyme activity in Tilapia, Oreochromis aureus

    SciTech Connect

    Low, K.W.; Sin, Y.M.

    1995-02-01

    Phagocytosis is an important defense mechanism against foreign pathogenic organisms. The cells involved are phagocytes which are comprised of peripheral blood monocytes (tissue macrophages) and polymorphonuclear (PMN) leucocytes. These cells can be activated by either particulate or soluble stimuli and undergo a respiratory burst from which several reactive oxygen species (ROS) can be formed. The reactive oxygen species and some hydrolases generated in the cells are the major antibacterial agents released during phagocytosis. Chemiluminescence (CL) is emitted, in vitro, from phagocytizing human PMN neutrophils. A similar CL response was also encountered in fish phagocytes. ROS was the causative agent of the CL emitted during in vitro phagocytosis. Phagocytic activity can be monitored by measuring the CL response of the phagocytes. Lysozyme is one of the potent hydrolases which are involved in the destruction of pathogens during phagocytosis. In fish, it was found predominantly in haematopoietic tissues, PMN leucocytes and moncytes. This enzyme has been shown to have antibacterial activity against several pathogens in fish. A combined oxidative and hydrolytic attack upon the engulfed pathogens allow phagocytes to kill infectious agents effectively. However, severe suppression or enhancement of these two functions caused by some exogenous factors may be detrimental to the host tissues. It has been reported that inorganic mercury could inhibit, in vitro, the respiratory burst and the microbicidal activities of human PMN leucocytes. It was also reported that increased in vitro release of lysozyme was found in mercury-treated human PMN leucocytes. However, such work has not been reported in fish. The aim of this research was to examine whether mercury could exert similar effects on the CL response in phagocytes and tissue lysozyme activity in fish after they were exposed to different concentrations of mercuric chloride over a period of 3 wks. 17 refs., 1 fig., 1 tab.

  15. Mercury toxicity in the shark (Squalus acanthias) rectal gland: apical CFTR chloride channels are inhibited by mercuric chloride.

    PubMed

    Ratner, Martha A; Decker, Sarah E; Aller, Stephen G; Weber, Gerhard; Forrest, John N

    2006-03-01

    In the shark rectal gland, basolateral membrane proteins have been suggested as targets for mercury. To examine the membrane polarity of mercury toxicity, we performed experiments in three preparations: isolated perfused rectal glands, primary monolayer cultures of rectal gland epithelial cells, and Xenopus oocytes expressing the shark cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel. In perfused rectal glands we observed: (1) a dose-dependent inhibition by mercury of forskolin/3-isobutyl-1-methylxanthine (IBMX)-stimulated chloride secretion; (2) inhibition was maximal when mercury was added before stimulation with forskolin/IBMX; (3) dithiothrietol (DTT) and glutathione (GSH) completely prevented inhibition of chloride secretion. Short-circuit current (Isc) measurements in monolayers of rectal gland epithelial cells were performed to examine the membrane polarity of this effect. Mercuric chloride inhibited Isc more potently when applied to the solution bathing the apical vs. the basolateral membrane (23 +/- 5% and 68 +/- 5% inhibition at 1 and 10 microM HgCl2 in the apical solution vs. 2 +/- 0.9% and 14 +/- 5% in the basolateral solution). This inhibition was prevented by pre-treatment with apical DTT or GSH; however, only the permeant reducing agent DTT reversed mercury inhibition when added after exposure. When the shark rectal gland CFTR channel was expressed in Xenopus oocytes and chloride conductance was measured by two-electrode voltage clamping, we found that 1 microM HgCl2 inhibited forskolin/IBMX conductance by 69.2 +/- 2.0%. We conclude that in the shark rectal gland, mercury inhibits chloride secretion by interacting with the apical membrane and that CFTR is the likely site of this action.

  16. Pharmacological characterization of the effects of methylmercury and mercuric chloride on spontaneous noradrenaline release from rat hippocampal slices.

    PubMed

    Gassó, S; Suñol, C; Sanfeliu, C; Rodríguez-Farré, E; Cristòfol, R M

    2000-01-01

    The environmental contaminants methylmercury (MeHg) and mercuric chloride (HgCl2) stimulated the spontaneous release of [3H]noradrenaline ([3H]NA) from hippocampal slices in a time- and concentration-dependent manner. Both MeHg and HgCl2 were similarly potent, with an EC50 of 88.4 microM and 75.9 microM, respectively. The releasing effects of MeHg and HgCl2 increased in the presence of desipramine, showing that the mechanism does not involve reversal of the transmitter transporter, and were completely blocked by reserpine preincubation, indicating a vesicular origin of [3H]NA release. The voltage-gated Na+ channel blocker tetrodotoxin (TTX) did not affect the response to mercury compounds. [3H]NA release elicited by MeHg was partially dependent on extracellular Ca2+, since it decreased significantly in a Ca2+-free EGTA-containing medium whereas HgCl2 induced a release of [3H]NA independent of extracellular Ca2+. Neither Ca2+-channels blockers, cobalt chloride (CoCl2) and (omega-conotoxin-GVIA, nor the Na+/Ca2+-exchanger inhibitor benzamil reduced MeHg-evoked [3H]NA release. Moreover, thapsigargin or caffeine, endoplasmic reticulum Ca2+-depletors, did not modify metal-evoked [3H]NA release, whereas ruthenium red, which inhibits the mitochondrial Ca2+ transport, decreased the effect of both MeHg and HgCl2. All these data indicate that, in hippocampal slices, mercury compounds release [3H]NA from the vesicular pool by a mechanism involving Ca2+ mobilization from mitochondrial stores.

  17. Effect of mercuric chloride on macrophage-mediated resistance mechanisms against infection with herpes simplex virus type 2.

    PubMed

    Ellermann-Eriksen, S; Christensen, M M; Mogensen, S C

    1994-11-11

    Macrophages play an important role in the early, nonspecific resistance to infection with herpes simplex virus. Mercuric chloride (HgCl2) accumulates in macrophages and has in certain concentrations a marked influence on the functional capacity of these cells. Therefore the influence of HgCl2 on resistance to generalized infection with herpes simplex virus type 2 (HSV-2) in mice and its effect on the HSV-2-induced activation of macrophages in vitro was examined. Mice injected intraperitoneally with HgCl2 24 h before infection with HSV-2 had more than 100 times higher virus titres in the liver 4 days after infection than mice not receiving any mercury. HgCl2 exerted a toxic effect on macrophages in vitro, which was especially pronounced during their adherence. Macrophages infected with HSV-2 were activated for an enhanced respiratory burst. This activation was abolished by treatment of the cells for 24 h with relatively low concentrations of HgCl2, resulting in macrophages with a potential to react with a respiratory burst comparable to that of uninfected cells. The HSV-2-induced activation of macrophages is mediated through the production and synergistic interaction of interferon-alpha/beta and tumour necrosis factor-alpha in an autocrine manner. The ability of these cytokines to activate macrophages and to interact synergistically was not affected by mercury. However the production by macrophages of both cytokines during the HSV-2 infection, but especially interferon-alpha/beta, which is essential for the activation, was reduced at low concentrations of HgCl2. Collectively these data indicate that mercury, by interfering with the early macrophage-production of cytokines, disables the early control of virus replication, leading to an enhanced infection.

  18. Purification and properties of Escherichia coli dimethyl sulfoxide reductase, an iron-sulfur molybdoenzyme with broad substrate specificity.

    PubMed

    Weiner, J H; MacIsaac, D P; Bishop, R E; Bilous, P T

    1988-04-01

    Dimethyl sulfoxide reductase, a terminal electron transfer enzyme, was purified from anaerobically grown Escherichia coli harboring a plasmid which codes for dimethyl sulfoxide reductase. The enzyme was purified to greater than 90% homogeneity from cell envelopes by a three-step purification procedure involving extraction with the detergent Triton X-100, chromatofocusing, and DEAE ion-exchange chromatography. The purified enzyme was composed of three subunits with molecular weights of 82,600, 23,600, and 22,700 as identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The native molecular weight was determined by gel electrophoresis to be 155,000. The purified enzyme contained 7.5 atoms of iron and 0.34 atom of molybdenum per mol of enzyme. The presence of molybdopterin cofactor in dimethyl sulfoxide reductase was identified by reconstitution of cofactor-deficient NADPH nitrate reductase activity from Neurospora crassa nit-I mutant and by UV absorption and fluorescence emission spectra. The enzyme displayed a very broad substrate specificity, reducing various N-oxide and sulfoxide compounds as well as chlorate and hydroxylamine.

  19. Transcripts of Anthocyanidin Reductase and Leucoanthocyanidin Reductase and Measurement of Catechin and Epicatechin in Tartary Buckwheat

    PubMed Central

    Kim, Yeon Bok; Thwe, Aye Aye; Kim, YeJi; Li, Xiaohua; Cho, Jin Woong; Park, Phun Bum; Valan Arasu, Mariadhas; Abdullah Al-Dhabi, Naif; Kim, Sun-Ju; Suzuki, Tastsuro; Hyun Jho, Kwang; Park, Sang Un

    2014-01-01

    Anthocyanidin reductase (ANR) and leucoanthocyanidin reductase (LAR) play an important role in the monomeric units biosynthesis of proanthocyanidins (PAs) such as catechin and epicatechin in several plants. The aim of this study was to clone ANR and LAR genes involved in PAs biosynthesis and examine the expression of these two genes in different organs under different growth conditions in two tartary buckwheat cultivars, Hokkai T8 and T10. Gene expression was carried out by quantitative real-time RT-PCR, and catechin and epicatechin content was analyzed by high performance liquid chromatography. The expression pattern of ANR and LAR did not match the accumulation pattern of PAs in different organs of two cultivars. Epicatechin content was the highest in the flowers of both cultivars and it was affected by light in only Hokkai T8 sprouts. ANR and LAR levels in tartary buckwheat might be regulated by different mechanisms for catechin and epicatechin biosynthesis under light and dark conditions. PMID:24605062

  20. Transcripts of anthocyanidin reductase and leucoanthocyanidin reductase and measurement of catechin and epicatechin in tartary buckwheat.

    PubMed

    Kim, Yeon Bok; Thwe, Aye Aye; Kim, Yeji; Li, Xiaohua; Cho, Jin Woong; Park, Phun Bum; Valan Arasu, Mariadhas; Abdullah Al-Dhabi, Naif; Kim, Sun-Ju; Suzuki, Tastsuro; Hyun Jho, Kwang; Park, Sang Un

    2014-01-01

    Anthocyanidin reductase (ANR) and leucoanthocyanidin reductase (LAR) play an important role in the monomeric units biosynthesis of proanthocyanidins (PAs) such as catechin and epicatechin in several plants. The aim of this study was to clone ANR and LAR genes involved in PAs biosynthesis and examine the expression of these two genes in different organs under different growth conditions in two tartary buckwheat cultivars, Hokkai T8 and T10. Gene expression was carried out by quantitative real-time RT-PCR, and catechin and epicatechin content was analyzed by high performance liquid chromatography. The expression pattern of ANR and LAR did not match the accumulation pattern of PAs in different organs of two cultivars. Epicatechin content was the highest in the flowers of both cultivars and it was affected by light in only Hokkai T8 sprouts. ANR and LAR levels in tartary buckwheat might be regulated by different mechanisms for catechin and epicatechin biosynthesis under light and dark conditions.

  1. Docking and molecular dynamics studies at trypanothione reductase and glutathione reductase active sites.

    PubMed

    Iribarne, Federico; Paulino, Margot; Aguilera, Sara; Murphy, Miguel; Tapia, Orlando

    2002-05-01

    A theoretical docking study on the active sites of trypanothione reductase (TR) and glutathione reductase (GR) with the corresponding natural substrates, trypanothione disulfide (T[S]2) and glutathione disulfide (GSSG), is reported. Molecular dynamics simulations were carried out in order to check the robustness of the docking results. The energetic results are in agreement with previous experimental findings and show the crossed complexes have lower stabilization energies than the natural ones. To test DOCK3.5, four nitro furanic compounds, previously designed as potentially active anti-chagasic molecules, were docked at the GR and TR active sites with the DOCK3.5 procedure. A good correlation was found between differential inhibitory activity and relative interaction energy (affinity). The results provide a validation test for the use of DOCK3.5 in connection with the design of anti-chagasic drugs.

  2. A high-throughput assay format for determination of nitrate reductase and nitrite reductase enzyme activities

    SciTech Connect

    McNally, N.; Liu, Xiang Yang; Choudary, P.V.

    1997-01-01

    The authors describe a microplate-based high-throughput procedure for rapid assay of the enzyme activities of nitrate reductase and nitrite reductase, using extremely small volumes of reagents. The new procedure offers the advantages of rapidity, small sample size-nanoliter volumes, low cost, and a dramatic increase in the throughput sample number that can be analyzed simultaneously. Additional advantages can be accessed by using microplate reader application software packages that permit assigning a group type to the wells, recording of the data on exportable data files and exercising the option of using the kinetic or endpoint reading modes. The assay can also be used independently for detecting nitrite residues/contamination in environmental/food samples. 10 refs., 2 figs.

  3. Expression in Escherichia coli of Cytochrome c Reductase Activity from a Maize NADH:Nitrate Reductase Complementary DNA 1

    PubMed Central

    Campbell, Wilbur H.

    1992-01-01

    A cDNA clone was isolated from a maize (Zea mays L. cv W64A×W183E) scutellum λgt11 library using maize leaf NADH:nitrate reductase Zmnr1 cDNA clone as a hybridization probe; it was designated Zmnr1S. Zmnr1S was shown to be an NADH:nitrate reductase clone by nucleotide sequencing and comparison of its deduced amino acid sequence to Zmnr1. Zmnr1S, which is 1.8 kilobases in length and contains the code for both the cytochrome b and flavin adenine dinucleotide domains of nitrate reductase, was cloned into the EcoRI site of the Escherichia coli expression vector pET5b and expressed. The cell lysate contained NADH:cytochrome c reductase activity, which is a characteristic partial activity of NADH:nitrate reductase dependent on the cytochrome b and flavin adenine dinucleotide domains. Recombinant cytochrome c reductase was purified by immunoaffinity chromatography on monoclonal antibody Zm2(69) Sepharose. The purified cytochrome c reductase, which had a major size of 43 kilodaltons, was inhibited by polyclonal antibodies for maize leaf NADH:nitrate reductase and bound these antibodies when blotted to nitrocellulose. Ultraviolet and visible spectra of oxidized and NADH-reduced recombinant cytochrome c reductase were nearly identical with those of maize leaf NADH:nitrate reductase. These two enzyme forms also had very similar kinetic properties with respect to NADH-dependent cytochrome c and ferricyanide reduction. ImagesFigure 2Figure 3 PMID:16668941

  4. Kinetic mechanism of pulmonary carbonyl reductase.

    PubMed

    Matsuura, K; Nakayama, T; Nakagawa, M; Hara, A; Sawada, H

    1988-05-15

    The kinetic mechanism of guinea-pig lung carbonyl reductase was studied at pH 7 in the forward reaction with five carbonyl substrates and NAD(P)H and in the reverse reaction with propan-2-ol and NAD(P)+. In each case the enzyme mechanism was sequential, and product-inhibition studies were consistent with a di-iso ordered bi bi mechanism, in which NAD(P)H binds to the enzyme first and NAD(P)+ leaves last and the binding of cofactor induces isomerization. The kinetic and binding studies of the cofactors and several inhibitors such as pyrazole, benzoic acid, Cibacron Blue and benzamide indicate that the cofactor and Cibacron Blue bind to the free enzyme whereas the other inhibitors bind to the binary and/or ternary complexes.

  5. Kinetic mechanism of pulmonary carbonyl reductase.

    PubMed Central

    Matsuura, K; Nakayama, T; Nakagawa, M; Hara, A; Sawada, H

    1988-01-01

    The kinetic mechanism of guinea-pig lung carbonyl reductase was studied at pH 7 in the forward reaction with five carbonyl substrates and NAD(P)H and in the reverse reaction with propan-2-ol and NAD(P)+. In each case the enzyme mechanism was sequential, and product-inhibition studies were consistent with a di-iso ordered bi bi mechanism, in which NAD(P)H binds to the enzyme first and NAD(P)+ leaves last and the binding of cofactor induces isomerization. The kinetic and binding studies of the cofactors and several inhibitors such as pyrazole, benzoic acid, Cibacron Blue and benzamide indicate that the cofactor and Cibacron Blue bind to the free enzyme whereas the other inhibitors bind to the binary and/or ternary complexes. PMID:3048244

  6. Methylenetetrahydrofolate reductase: biochemical characterization and medical significance.

    PubMed

    Trimmer, Elizabeth E

    2013-01-01

    Methylenetetrahydrofolate reductase (MTHFR) catalyzes the reduction of 5,10-methylenetetrahydofolate (CH2-H4folate) to 5-methyltetrahydrofolate (CH3-H4folate). The enzyme employs a noncovalently-bound flavin adenine dinucleotide (FAD), which accepts reducing equivalents from NAD(P)H and transfers them to CH2-H4folate. The reaction provides the sole source of CH3-H4folate, which is utilized by methionine synthase in the synthesis of methionine from homocysteine. MTHFR plays a key role in folate metabolism and in the homeostasis of homocysteine; mutations in the enzyme lead to hyperhomocyst(e)inemia. A common C677T polymorphism in MTHFR has been associated with an increased risk for the development of cardiovascular disease, Alzheimer's disease, and depression in adults, and of neural tube defects in the fetus. The mutation also confers protection for certain types of cancers. This review presents the current knowledge of the enzyme, its biochemical characterization, and medical significance.

  7. The cytochrome bd respiratory oxygen reductases.

    PubMed

    Borisov, Vitaliy B; Gennis, Robert B; Hemp, James; Verkhovsky, Michael I

    2011-11-01

    Cytochrome bd is a respiratory quinol: O₂ oxidoreductase found in many prokaryotes, including a number of pathogens. The main bioenergetic function of the enzyme is the production of a proton motive force by the vectorial charge transfer of protons. The sequences of cytochromes bd are not homologous to those of the other respiratory oxygen reductases, i.e., the heme-copper oxygen reductases or alternative oxidases (AOX). Generally, cytochromes bd are noteworthy for their high affinity for O₂ and resistance to inhibition by cyanide. In E. coli, for example, cytochrome bd (specifically, cytochrome bd-I) is expressed under O₂-limited conditions. Among the members of the bd-family are the so-called cyanide-insensitive quinol oxidases (CIO) which often have a low content of the eponymous heme d but, instead, have heme b in place of heme d in at least a majority of the enzyme population. However, at this point, no sequence motif has been identified to distinguish cytochrome bd (with a stoichiometric complement of heme d) from an enzyme designated as CIO. Members of the bd-family can be subdivided into those which contain either a long or a short hydrophilic connection between transmembrane helices 6 and 7 in subunit I, designated as the Q-loop. However, it is not clear whether there is a functional consequence of this difference. This review summarizes current knowledge on the physiological functions, genetics, structural and catalytic properties of cytochromes bd. Included in this review are descriptions of the intermediates of the catalytic cycle, the proposed site for the reduction of O₂, evidence for a proton channel connecting this active site to the bacterial cytoplasm, and the molecular mechanism by which a membrane potential is generated. 2011 Elsevier B.V. All rights reserved.

  8. The cytochrome bd respiratory oxygen reductases

    PubMed Central

    Borisov, Vitaliy B.; Gennis, Robert B.; Hemp, James; Verkhovsky, Michael I.

    2011-01-01

    Summary Cytochrome bd is a respiratory quinol:O2 oxidoreductase found in many prokaryotes, including a number of pathogens. The main bioenergetic function of the enzyme is the production of a proton motive force by the vectorial charge transfer of protons. The sequences of cytochromes bd are not homologous to those of the other respiratory oxygen reductases, i.e., the heme-copper oxygen reductases or alternative oxidases (AOX). Generally, cytochromes bd are noteworthy for their high affinity for O2 and resistance to inhibition by cyanide. In E. coli, for example, cytochrome bd (specifically, cytochrome bd-I) is expressed under O2-limited conditions. Among the members of the bd-family are the so-called cyanide-insensitive quinol oxidases (CIO) which often have a low content of the eponymous heme d but, instead, have heme b in place of heme d in at least a majority of the enzyme population. However, at this point, no sequence motif has been identified to distinguish cytochrome bd (with a stoichiometric complement of heme d) from an enzyme designated as CIO. Members of the bd-family can be subdivided into those which contain either a long or a short hydrophilic connection between transmembrane helices 6 and 7 in subunit I, designated as the Q-loop. However, it is not clear whether there is a functional consequence of this difference. This review summarizes current knowledge on the physiological functions, genetics, structural and catalytic properties of cytochromes bd. Included in this review are descriptions of the intermediates of the catalytic cycle, the proposed site for the reduction of O2, evidence for a proton channel connecting this active site to the bacterial cytoplasm, and the molecular mechanism by which a membrane potential is generated. PMID:21756872

  9. Ameliorating effect of β-carotene on antioxidant response and hematological parameters of mercuric chloride toxicity in Nile tilapia (Oreochromis niloticus).

    PubMed

    Elseady, Y; Zahran, E

    2013-08-01

    The impact of different levels of dietary β-carotene to alleviate the effect of mercuric chloride toxicity in Nile tilapia was assessed. Semi-purified diets containing 0, 40, and 100 mg β-carotene kg(-1) dry diet were fed for 21 days, which were subjected to sublethal concentration of mercuric chloride (0.05 ppm). Hematological and biochemical parameters, lipid profile, and antioxidant response were examined. All hematological parameters of tilapia fish starting from second week of toxicity were significantly decreased. A significant increasing trend in liver enzymes (ALT and AST) were observed parallel to the time of toxicity and peroxide radicals (MDA) appearing significantly increased in toxicated group without carotene supplement, although carotene supplementation return all parameters within the control levels. Mercury accumulated significantly in fish liver and white muscles in toxicated group while it showed a significant reduction in dietary β-carotene-treated group. Overall, it can be used as immunostimulant and alleviate the suppression effect resulted from immune depressive stressful condition in farmed Nile tilapia.

  10. Modifications in rat testicular morphology and increases in IFN-gamma serum levels by the oral administration of subtoxic doses of mercuric chloride.

    PubMed

    Penna, Salvador; Pocino, Marisol; Marval, Maria Josefina; Lloreta, José; Gallardo, Luis; Vila, Joan

    2009-01-01

    Mercury induces structural and functional damage in several organs, however the effects of subtoxic doses of the metal on the male reproductive system are not well defined. In order to analyze testicular and epididymal morphological alterations and changes in IL-4 or IFN-gamma serum levels, adult male Sprague-Dawley rats received 0.01, 0.05 or 0.1 microg/ml of mercuric chloride (HgCl(2)) in deionized water for 1 to 7 months by oral route. Controls received deionized water alone. Twenty rats, separated in four groups of five animals each, were used per time of exposure. Progressive degenerative lesions consisting of lack of germ cell cohesion and desquamation, arrest at spermatocyte stage and hypospermatogenesis were observed in seminiferous epithelium by light and electron microscopy. Leydig cells showed cytoplasmic vacuolation and nuclear signs of cell death. Loss of peritubular cell aggregation was evidenced in the epididymis. Mercury accumulation was detected in both organs by mass spectroscopy. Rats showed enhanced IFN-gamma serum levels as compared to controls but only reached significance after 7 months of mercury administration. Subtoxic doses of inorganic mercury could lead to reproductive and immunological alterations. The results demonstrate that sublethal concentrations of mercuric chloride are enough to induce morphological and ultrastructural modifications in male reproductive organs. These contribute to functional alterations of spermatogenesis with arrest at spermatocyte stage, hypospermatogenesis and possibly impaired steroidogenesis which together could affect male fertility.

  11. Analytical interferences of mercuric chloride preservative in environmental water samples: Determination of organic compounds isolated by continuous liquid-liquid extraction or closed-loop stripping

    USGS Publications Warehouse

    Foreman, W.T.; Zaugg, S.D.; Falres, L.M.; Werner, M.G.; Leiker, T.J.; Rogerson, P.F.

    1992-01-01

    Analytical interferences were observed during the determination of organic compounds in groundwater samples preserved with mercuric chloride. The nature of the interference was different depending on the analytical isolation technique employed. (1) Water samples extracted with dichloromethane by continuous liquid-liquid extraction (CLLE) and analyzed by gas chromatography/mass spectrometry revealed a broad HgCl2 'peak' eluting over a 3-5-min span which interfered with the determination of coeluting organic analytes. Substitution of CLLE for separatory funnel extraction in EPA method 508 also resulted in analytical interferences from the use of HgCl2 preservative. (2) Mercuric chloride was purged, along with organic contaminants, during closed-loop stripping (CLS) of groundwater samples and absorbed onto the activated charcoal trap. Competitive sorption of the HgCl2 by the trap appeared to contribute to the observed poor recoveries for spiked organic contaminants. The HgCl2 was not displaced from the charcoal with the dichloromethane elution solvent and required strong nitric acid to achieve rapid, complete displacement. Similar competitive sorption mechanisms might also occur in other purge and trap methods when this preservative is used.

  12. Enhanced silver nanoparticle synthesis by optimization of nitrate reductase activity.

    PubMed

    Vaidyanathan, Ramanathan; Gopalram, Shubaash; Kalishwaralal, Kalimuthu; Deepak, Venkataraman; Pandian, Sureshbabu Ram Kumar; Gurunathan, Sangiliyandi

    2010-01-01

    Nanostructure materials are attracting a great deal of attention because of their potential for achieving specific processes and selectivity, especially in biological and pharmaceutical applications. The generation of silver nanoparticles using optimized nitrate reductase for the reduction of Ag(+) with the retention of enzymatic activity in the complex is being reported. This report involves the optimization of enzyme activity to bring about enhanced nanoparticle synthesis. Response surface methodology and central composite rotary design (CCRD) were employed to optimize a fermentation medium for the production of nitrate reductase by Bacillus licheniformis at pH 8. The four variables involved in the study of nitrate reductase were Glucose, Peptone, Yeast extract and KNO(3). Glucose had a significant effect on nitrate reductase production. The optimized medium containing (%) Glucose: 1.5, Peptone: 1, Yeast extract: 0.35 and KNO(3): 0.35 resulted in a nitrate reductase activity of 452.206 U/ml which is same as that of the central level. The medium A (showing least nitrate reductase activity) and the medium B (showing maximum nitrate reductase activity) were compared for the synthesis. Spectrophotometric analysis revealed that the particles exhibited a peak at 431 nm and the A(431) for the medium B was 2-fold greater than that of the medium A. The particles were also characterized using TEM. The particles synthesized using the optimized enzyme activity ranged from 10 to 80 nm and therefore can be extended to various medicinal applications.

  13. Toxicology and Carcinogenesis Studies of Mercuric Chloride (CAS No. 7487-94-7) in F344 Rats and B6C3F1 Mice (Gavage Studies).

    PubMed

    1993-02-01

    Mercuric chloride is an inorganic compound that has been used in agriculture as a fungicide, in medicine as a topical antiseptic and disinfectant, and in chemistry as an intermediate in the production of other mercury compounds. The widespread use of mercury has resulted in increased levels of mercury in rivers and lakes. Mercuric chloride was evaluated in toxicity and carcinogenicity studies because of its extensive use and its occurrence as an environmental pollutant, and because of the lack of adequate long-term rodent studies. Toxicology and carcinogenesis studies were conducted by administering mercuric chloride (greater than 99% pure) in deionized water by gavage to groups of F344 rats or B6C3F1 mice for 16 days, 6 months, and 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium (strains TA98, TA100, TA1535, and TA1537), in mouse lymphoma L5178Y cells, in Chinese hamster ovary cells, and in Drosophila melanogaster. 16-DAY STUDIES: Groups of five rats of each sex received 0, 1.25, 2.5, 5, 10, or 20 mg mercuric chloride/kg body weight and groups of five mice of each sex received 0, 5, 10, 20, 40, or 80 mg/kg in deionized water by gavage for 12 dose days. Two male rats in the 20 mg/kg group died in the first week, as did all male and four female mice from the 80 mg/kg group and one male mouse from the 40 mg/kg group. The final mean body weight of male rats receiving 20 mg/kg was 10% lower than that of the controls; the final mean body weight of 20 mg/kg females was 9% lower than that of the controls. Final mean body weights and body weight gains of dosed mice were similar to those of the controls. Absolute and relative kidney weights of male rats receiving 2.5 mg/kg or greater doses and of female rats administered 5 mg/kg or more were significantly greater than those of the controls. Absolute kidney weights of mice were significantly increased in all male dose groups and in the 40 mg/kg female dose group; relative kidney weights of dosed

  14. FAD binding, cobinamide binding and active site communication in the corrin reductase (CobR)

    PubMed Central

    Lawrence, Andrew D.; Taylor, Samantha L.; Scott, Alan; Rowe, Michelle L.; Johnson, Christopher M.; Rigby, Stephen E. J.; Geeves, Michael A.; Pickersgill, Richard W.; Howard, Mark J.; Warren, Martin J.

    2014-01-01

    Adenosylcobalamin, the coenzyme form of vitamin B12, is one Nature's most complex coenzyme whose de novo biogenesis proceeds along either an anaerobic or aerobic metabolic pathway. The aerobic synthesis involves reduction of the centrally chelated cobalt metal ion of the corrin ring from Co(II) to Co(I) before adenosylation can take place. A corrin reductase (CobR) enzyme has been identified as the likely agent to catalyse this reduction of the metal ion. Herein, we reveal how Brucella melitensis CobR binds its coenzyme FAD (flavin dinucleotide) and we also show that the enzyme can bind a corrin substrate consistent with its role in reduction of the cobalt of the corrin ring. Stopped-flow kinetics and EPR reveal a mechanistic asymmetry in CobR dimer that provides a potential link between the two electron reduction by NADH to the single electron reduction of Co(II) to Co(I). PMID:24909839

  15. Effect of mercuric chloride on the kinetics of cationic and substrate activation of the rat brain microsomal ATPase system.

    PubMed

    Rajanna, B; Chetty, C S; Rajanna, S

    1990-06-15

    Mercuric chloride (HgCl2), a neurotoxic compound, inhibited the adenosine triphosphatase (ATPase) system in a concentration-dependent manner. Hydrolysis of ATP was linear with time with or without HgCl2 in the reaction mixtures. Higher inhibition of (Na(+)-K+)ATPase activity by HgCl2 was observed in alkaline (8.0 to 9.0) pH and at lower temperatures (17 to 32 degrees). Activation energy values were increased slightly in the presence of HgCl2. Activation of (Na(+)-K+)ATPase by ATP in the presence of HgCl2 showed a decrease in Vmax from 15.29 to 5.0 mumol of inorganic phosphate (Pi)/mg protein/hr with no change in Km. Similarly, activation of K(+)-stimulated p-nitrophenyl phosphatase (K(+)-PNPPase) in the presence of HgCl2 showed a decrease in Vmax from 3.26 to 1.35 mumols of p-nitrophenol (PNP)/mg protein/hr with no change in Km. K(+)-activation kinetic studies indicated that HgCl2 decreased Vmax from 14.01 to 4.30 mumols Pi/mg protein/hr in the case of (Na(+)-K+)ATPase and from 3.45 to 2.40 mumols PNP/mg protein/hr in the case of K(+)-PNPPase with no changes in Km. Na(+)-activation of (Na(+)-K+)ATPase in the presence of HgCl2 showed a decrease in Vmax from 11.06 to 3.23 mumols Pi/mg protein/hr and an increase in Km from 1.06 to 2.08 mM. Preincubation of microsomes with sulfhydryl (SH) agents dithiothreitol, cysteine and glutathione protected HgCl2-inhibition of (Na(+)-K+)ATPase. The data suggest that HgCl2 inhibited (Na(+)-K+)ATPase by interfering with the dephosphorylation of the enzyme-phosphoryl complex.

  16. Carboxylation mechanism and stereochemistry of crotonyl-CoA carboxylase/reductase, a carboxylating enoyl-thioester reductase

    PubMed Central

    Erb, Tobias J.; Brecht, Volker; Fuchs, Georg; Müller, Michael; Alber, Birgit E.

    2009-01-01

    Chemo- and stereoselective reductions are important reactions in chemistry and biology, and reductases from biological sources are increasingly applied in organic synthesis. In contrast, carboxylases are used only sporadically. We recently described crotonyl-CoA carboxylase/reductase, which catalyzes the reduction of (E)-crotonyl-CoA to butyryl-CoA but also the reductive carboxylation of (E)-crotonyl-CoA to ethylmalonyl-CoA. In this study, the complete stereochemical course of both reactions was investigated in detail. The pro-(4R) hydrogen of NADPH is transferred in both reactions to the re face of the C3 position of crotonyl-CoA. In the course of the carboxylation reaction, carbon dioxide is incorporated in anti fashion at the C2 atom of crotonyl-CoA. For the reduction reaction that yields butyryl-CoA, a solvent proton is added in anti fashion instead of the CO2. Amino acid sequence analysis showed that crotonyl-CoA carboxylase/reductase is a member of the medium-chain dehydrogenase/reductase superfamily and shares the same phylogenetic origin. The stereospecificity of the hydride transfer from NAD(P)H within this superfamily is highly conserved, although the substrates and reduction reactions catalyzed by its individual representatives differ quite considerably. Our findings led to a reassessment of the stereospecificity of enoyl(-thioester) reductases and related enzymes with respect to their amino acid sequence, revealing a general pattern of stereospecificity that allows the prediction of the stereochemistry of the hydride transfer for enoyl reductases of unknown specificity. Further considerations on the reaction mechanism indicated that crotonyl-CoA carboxylase/reductase may have evolved from enoyl-CoA reductases. This may be useful for protein engineering of enoyl reductases and their application in biocatalysis. PMID:19458256

  17. The structure of apo and holo forms of xylose reductase, a dimeric aldo-keto reductase from Candida tenuis.

    PubMed

    Kavanagh, Kathryn L; Klimacek, Mario; Nidetzky, Bernd; Wilson, David K

    2002-07-16

    Xylose reductase is a homodimeric oxidoreductase dependent on NADPH or NADH and belongs to the largely monomeric aldo-keto reductase superfamily of proteins. It catalyzes the first step in the assimilation of xylose, an aldose found to be a major constituent monosaccharide of renewable plant hemicellulosic material, into yeast metabolic pathways. It does this by reducing open chain xylose to xylitol, which is reoxidized to xylulose by xylitol dehydrogenase and metabolically integrated via the pentose phosphate pathway. No structure has yet been determined for a xylose reductase, a dimeric aldo-keto reductase or a family 2 aldo-keto reductase. The structures of the Candida tenuis xylose reductase apo- and holoenzyme, which crystallize in spacegroup C2 with different unit cells, have been determined to 2.2 A resolution and an R-factor of 17.9 and 20.8%, respectively. Residues responsible for mediating the novel dimeric interface include Asp-178, Arg-181, Lys-202, Phe-206, Trp-313, and Pro-319. Alignments with other superfamily members indicate that these interactions are conserved in other dimeric xylose reductases but not throughout the remainder of the oligomeric aldo-keto reductases, predicting alternate modes of oligomerization for other families. An arrangement of side chains in a catalytic triad shows that Tyr-52 has a conserved function as a general acid. The loop that folds over the NAD(P)H cosubstrate is disordered in the apo form but becomes ordered upon cosubstrate binding. A slow conformational isomerization of this loop probably accounts for the observed rate-limiting step involving release of cosubstrate. Xylose binding (K(m) = 87 mM) is mediated by interactions with a binding pocket that is more polar than a typical aldo-keto reductase. Modeling of xylose into the active site of the holoenzyme using ordered waters as a guide for sugar hydroxyls suggests a convincing mode of substrate binding.

  18. Solubilization and Resolution of the Membrane-Bound Nitrite Reductase from Paracoccus Halodenitrificans into Nitrite and Nitric Oxide Reductases

    NASA Technical Reports Server (NTRS)

    Grant, Michael A.; Cronin, Sonja E.; Hochstein, Lawrence I.

    1984-01-01

    Membranes prepared from Paracoccus halodenitrificans reduced nitrite or nitric oxide to nitrous oxide. Extraction of these membranes with the detergent CHAPSO [3-(3-Chlolamidoporopyldimethylammonio)-1-(2- hydroxy-1-propanesulfonate)], followed by ammonium sulfate fractionation of the solubilized proteins, resulted in the separation of nitrite and nitric oxide reductase activities. The fraction containing nitrite reductase activity spectrally resembled a cd-type cytochrome. Several cytochromes were detected in the nitric oxide reductase fraction. Which, if any, of these cytochromes is associated with the reduction of nitric oxide is not clear at this time.

  19. Solubilization and Resolution of the Membrane-Bound Nitrite Reductase from Paracoccus Halodenitrificans into Nitrite and Nitric Oxide Reductases

    NASA Technical Reports Server (NTRS)

    Grant, Michael A.; Cronin, Sonja E.; Hochstein, Lawrence I.

    1984-01-01

    Membranes prepared from Paracoccus halodenitrificans reduced nitrite or nitric oxide to nitrous oxide. Extraction of these membranes with the detergent CHAPSO [3-(3-Chlolamidoporopyldimethylammonio)-1-(2- hydroxy-1-propanesulfonate)], followed by ammonium sulfate fractionation of the solubilized proteins, resulted in the separation of nitrite and nitric oxide reductase activities. The fraction containing nitrite reductase activity spectrally resembled a cd-type cytochrome. Several cytochromes were detected in the nitric oxide reductase fraction. Which, if any, of these cytochromes is associated with the reduction of nitric oxide is not clear at this time.

  20. Comparative Efficacy of Pre-feeding, Post-feeding and Combined Pre- and Post-feeding of Two Microdoses of a Potentized Homeopathic Drug, Mercurius Solubilis, in Ameliorating Genotoxic Effects Produced by Mercuric Chloride in Mice

    PubMed Central

    2004-01-01

    Mercury and its derivatives have become an alarming environmental problem, necessitating the search for effective antagonists, including homeopathic drugs, which are generally used in micro doses and are devoid of any palpable side-effects. On the basis of homeopathic similia principle, two potencies of Mercurius solubilis (Merc Sol-30 and Merc Sol-200) were tested by three administrative modes, i.e. pre-feeding, post-feeding and combined pre- and post-feeding, for their possible efficacy in ameliorating mercuric chloride-induced genotoxicity in mice. Healthy mice, Mus musculus, were intraperitoneally injected with 0.06% solution of mercuric chloride at the rate of 1 ml/100 g of body weight, and assessed for genotoxic effects through conventional endpoints. i.e. chromosome aberrations, micronuclei, mitotic index and sperm head abnormality, keeping suitable controls. Mercuric chloride-treated mice were divided into three sub-groups, which were orally administered with the drug prior to, after and both prior to and after injection of mercuric chloride, and their genotoxic effects were analysed at specific intervals of fixation. Mercuric chloride treatment generally produced more chromosome aberations, micronuclei and sperm head anomaly in mice, but the mitotic index appeared to be slightly reduced. While chromosome aberations, micronuclei and sperm head anomaly were generally reduced in the drug-fed series, the mitotic index showed an apparent increase. In most cases, the combined pre- and post-feeding mode appeared to show the maximum amelioration, followed by post-feeding and pre-feeding, in that order. The amelioration by Merc Sol-200 appeared to be slightly more pronounced. We conclude that potentized homeopathic drugs can serve as possible anti-genotoxic agents against specific environmental mutagens, including toxic heavy metals. PMID:15864357

  1. Exploration of Nitrate Reductase Metabolic Pathway in Corynebacterium pseudotuberculosis

    PubMed Central

    Abreu, Vinícius; Diniz, Carlos; Dorneles, Elaine M. S.; Barh, Debmalya

    2017-01-01

    Based on the ability of nitrate reductase synthesis, Corynebacterium pseudotuberculosis is classified into two biovars: Ovis and Equi. Due to the presence of nitrate reductase, the Equi biovar can survive in absence of oxygen. On the other hand, Ovis biovar that does not have nitrate reductase is able to adapt to various ecological niches and can grow on certain carbon sources. Apart from these two biovars, some other strains are also able to carry out the reduction of nitrate. The enzymes that are involved in electron transport chain are also identified by in silico methods. Findings about pathogen metabolism can contribute to the identification of relationship between nitrate reductase and the C. pseudotuberculosis pathogenicity, virulence factors, and discovery of drug targets. PMID:28316974

  2. Exploration of Nitrate Reductase Metabolic Pathway in Corynebacterium pseudotuberculosis.

    PubMed

    Almeida, Sintia; Sousa, Cassiana; Abreu, Vinícius; Diniz, Carlos; Dorneles, Elaine M S; Lage, Andrey P; Barh, Debmalya; Azevedo, Vasco

    2017-01-01

    Based on the ability of nitrate reductase synthesis, Corynebacterium pseudotuberculosis is classified into two biovars: Ovis and Equi. Due to the presence of nitrate reductase, the Equi biovar can survive in absence of oxygen. On the other hand, Ovis biovar that does not have nitrate reductase is able to adapt to various ecological niches and can grow on certain carbon sources. Apart from these two biovars, some other strains are also able to carry out the reduction of nitrate. The enzymes that are involved in electron transport chain are also identified by in silico methods. Findings about pathogen metabolism can contribute to the identification of relationship between nitrate reductase and the C. pseudotuberculosis pathogenicity, virulence factors, and discovery of drug targets.

  3. Enantioselective imine reduction catalyzed by imine reductases and artificial metalloenzymes.

    PubMed

    Gamenara, Daniela; Domínguez de María, Pablo

    2014-05-21

    Adding value to organic synthesis. Novel imine reductases enable the enantioselective reduction of imines to afford optically active amines. Likewise, novel bioinspired artificial metalloenzymes can perform the same reaction as well. Emerging proof-of-concepts are herein discussed.

  4. Anti-HMG-CoA Reductase, Antioxidant, and Anti-Inflammatory Activities of Amaranthus viridis Leaf Extract as a Potential Treatment for Hypercholesterolemia

    PubMed Central

    Salvamani, Shamala; Gunasekaran, Baskaran; Shukor, Mohd Yunus; Shaharuddin, Noor Azmi; Sabullah, Mohd Khalizan

    2016-01-01

    Inflammation and oxidative stress are believed to contribute to the pathology of several chronic diseases including hypercholesterolemia (elevated levels of cholesterol in blood) and atherosclerosis. HMG-CoA reductase inhibitors of plant origin are needed as synthetic drugs, such as statins, which are known to cause adverse effects on the liver and muscles. Amaranthus viridis (A. viridis) has been used from ancient times for its supposedly medically beneficial properties. In the current study, different parts of A. viridis (leaf, stem, and seed) were evaluated for potential anti-HMG-CoA reductase, antioxidant, and anti-inflammatory activities. The putative HMG-CoA reductase inhibitory activity of A. viridis extracts at different concentrations was determined spectrophotometrically by NADPH oxidation, using HMG-CoA as substrate. A. viridis leaf extract revealed the highest HMG-CoA reductase inhibitory effect at about 71%, with noncompetitive inhibition in Lineweaver-Burk plot analysis. The leaf extract showed good inhibition of hydroperoxides, 2,2-diphenyl-1-picrylhydrazyl (DPPH), nitric oxide (NO), and ferric ion radicals in various concentrations. A. viridis leaf extract was proven to be an effective inhibitor of hyaluronidase, lipoxygenase, and xanthine oxidase enzymes. The experimental data suggest that A. viridis leaf extract is a source of potent antioxidant and anti-inflammatory agent and may modulate cholesterol metabolism by inhibition of HMG-CoA reductase. PMID:27051453

  5. 1,8-Dihydroxynaphthalene (DHN)-Melanin Biosynthesis Inhibitors Increase Erythritol Production in Torula corallina, and DHN-Melanin Inhibits Erythrose Reductase

    PubMed Central

    Lee, Jung-Kul; Jung, Hyung-Moo; Kim, Sang-Yong

    2003-01-01

    The yeast Torula corallina is a strong erythritol producer that is used in the industrial production of erythritol. However, melanin accumulation during culture represents a serious problem for the purification of erythritol from the fermentation broth. Melanin biosynthesis inhibitors such as 3,4-dihydroxyphenylalanine and 1,8-dihydroxynaphthalene (DHN)-melanin inhibitors were added to the T. corallina cultures. Only the DHN-melanin inhibitors showed an effect on melanin production, which suggests that the melanin formed during the culturing of T. corallina is derived from DHN. This finding was confirmed by the detection of a shunt product of the pentaketide pathway, flaviolin, and elemental analysis. Among the DHN-melanin inhibitors, tricyclazole was the most effective. Supplementation with tricyclazole enhanced the production of erythritol while significantly inhibiting the production of DHN-melanin and DHN-melanin biosynthetic enzymes, such as trihydroxynaphthalene reductase. The erythrose reductase from T. corallina was purified to homogeneity by ion-exchange and affinity chromatography. Purified erythrose reductase was significantly inhibited in vitro in a noncompetitive manner by elevated levels of DHN-melanin. In contrast, the level of erythrose reductase activity was unaffected by increasing concentrations of tricyclazole. These results suggest that supplemental tricyclazole reduces the production of DHN-melanin, which may lead to a reduction in the inhibition of erythrose reductase and a higher yield of erythritol. This is the first report to demonstrate that melanin biosynthesis inhibitors increase the production of a sugar alcohol in T. corallina. PMID:12788746

  6. Anti-HMG-CoA Reductase, Antioxidant, and Anti-Inflammatory Activities of Amaranthus viridis Leaf Extract as a Potential Treatment for Hypercholesterolemia.

    PubMed

    Salvamani, Shamala; Gunasekaran, Baskaran; Shukor, Mohd Yunus; Shaharuddin, Noor Azmi; Sabullah, Mohd Khalizan; Ahmad, Siti Aqlima

    2016-01-01

    Inflammation and oxidative stress are believed to contribute to the pathology of several chronic diseases including hypercholesterolemia (elevated levels of cholesterol in blood) and atherosclerosis. HMG-CoA reductase inhibitors of plant origin are needed as synthetic drugs, such as statins, which are known to cause adverse effects on the liver and muscles. Amaranthus viridis (A. viridis) has been used from ancient times for its supposedly medically beneficial properties. In the current study, different parts of A. viridis (leaf, stem, and seed) were evaluated for potential anti-HMG-CoA reductase, antioxidant, and anti-inflammatory activities. The putative HMG-CoA reductase inhibitory activity of A. viridis extracts at different concentrations was determined spectrophotometrically by NADPH oxidation, using HMG-CoA as substrate. A. viridis leaf extract revealed the highest HMG-CoA reductase inhibitory effect at about 71%, with noncompetitive inhibition in Lineweaver-Burk plot analysis. The leaf extract showed good inhibition of hydroperoxides, 2,2-diphenyl-1-picrylhydrazyl (DPPH), nitric oxide (NO), and ferric ion radicals in various concentrations. A. viridis leaf extract was proven to be an effective inhibitor of hyaluronidase, lipoxygenase, and xanthine oxidase enzymes. The experimental data suggest that A. viridis leaf extract is a source of potent antioxidant and anti-inflammatory agent and may modulate cholesterol metabolism by inhibition of HMG-CoA reductase.

  7. Purification and characterization of assimilatory nitrite reductase from Candida utilis.

    PubMed

    Sengupta, S; Shaila, M S; Rao, G R

    1996-07-01

    Nitrate assimilation in many plants, algae, yeasts and bacteria is mediated by two enzymes, nitrate reductase (EC 1.6.6.2) and nitrite reductase (EC 1.7.7.1). They catalyse the stepwise reduction of nitrate to nitrite and nitrite to ammonia respectively. The nitrite reductase from an industrially important yeast, Candida utilis, has been purified to homogeneity. Purified nitrite reductase is a heterodimer and the molecular masses of the two subunits are 58 and 66 kDa. The native enzyme exhibits a molecular mass of 126 kDa as analysed by gel filtration. The identify of the two subunits of nitrite reductase was confirmed by immunoblotting using antibody for Cucurbita pepo leaf nitrite reductase. The presence of two different sized transcripts coding for the two subunits was confirmed by (a) in vitro translation of mRNA from nitrate-induced C. utilis followed by immunoprecipitation of the in vitro translated products with heterologous nitrite reductase antibody and (b) Northern-blot analysis. The 66 kDa subunit is acidic in nature which is probably due to its phosphorylated status. The enzyme is stable over a range of temperatures. Both subunits can catalyse nitrite reduction, and the reconstituted enzyme, at a higher protein concentration, shows an activity similar to that of the purified enzyme. Each of these subunits has been shown to contain a few unique peptides in addition to a large number of common peptides. Reduced Methyl Viologen has been found to be as effective an electron donor as NADPH in the catalytic process, a phenomenon not commonly seen for nitrite reductases from other systems.

  8. Comparative anatomy of the aldo-keto reductase superfamily.

    PubMed Central

    Jez, J M; Bennett, M J; Schlegel, B P; Lewis, M; Penning, T M

    1997-01-01

    The aldo-keto reductases metabolize a wide range of substrates and are potential drug targets. This protein superfamily includes aldose reductases, aldehyde reductases, hydroxysteroid dehydrogenases and dihydrodiol dehydrogenases. By combining multiple sequence alignments with known three-dimensional structures and the results of site-directed mutagenesis studies, we have developed a structure/function analysis of this superfamily. Our studies suggest that the (alpha/beta)8-barrel fold provides a common scaffold for an NAD(P)(H)-dependent catalytic activity, with substrate specificity determined by variation of loops on the C-terminal side of the barrel. All the aldo-keto reductases are dependent on nicotinamide cofactors for catalysis and retain a similar cofactor binding site, even among proteins with less than 30% amino acid sequence identity. Likewise, the aldo-keto reductase active site is highly conserved. However, our alignments indicate that variation ofa single residue in the active site may alter the reaction mechanism from carbonyl oxidoreduction to carbon-carbon double-bond reduction, as in the 3-oxo-5beta-steroid 4-dehydrogenases (Delta4-3-ketosteroid 5beta-reductases) of the superfamily. Comparison of the proposed substrate binding pocket suggests residues 54 and 118, near the active site, as possible discriminators between sugar and steroid substrates. In addition, sequence alignment and subsequent homology modelling of mouse liver 17beta-hydroxysteroid dehydrogenase and rat ovary 20alpha-hydroxysteroid dehydrogenase indicate that three loops on the C-terminal side of the barrel play potential roles in determining the positional and stereo-specificity of the hydroxysteroid dehydrogenases. Finally, we propose that the aldo-keto reductase superfamily may represent an example of divergent evolution from an ancestral multifunctional oxidoreductase and an example of convergent evolution to the same active-site constellation as the short

  9. Comparative anatomy of the aldo-keto reductase superfamily.

    PubMed

    Jez, J M; Bennett, M J; Schlegel, B P; Lewis, M; Penning, T M

    1997-09-15

    The aldo-keto reductases metabolize a wide range of substrates and are potential drug targets. This protein superfamily includes aldose reductases, aldehyde reductases, hydroxysteroid dehydrogenases and dihydrodiol dehydrogenases. By combining multiple sequence alignments with known three-dimensional structures and the results of site-directed mutagenesis studies, we have developed a structure/function analysis of this superfamily. Our studies suggest that the (alpha/beta)8-barrel fold provides a common scaffold for an NAD(P)(H)-dependent catalytic activity, with substrate specificity determined by variation of loops on the C-terminal side of the barrel. All the aldo-keto reductases are dependent on nicotinamide cofactors for catalysis and retain a similar cofactor binding site, even among proteins with less than 30% amino acid sequence identity. Likewise, the aldo-keto reductase active site is highly conserved. However, our alignments indicate that variation ofa single residue in the active site may alter the reaction mechanism from carbonyl oxidoreduction to carbon-carbon double-bond reduction, as in the 3-oxo-5beta-steroid 4-dehydrogenases (Delta4-3-ketosteroid 5beta-reductases) of the superfamily. Comparison of the proposed substrate binding pocket suggests residues 54 and 118, near the active site, as possible discriminators between sugar and steroid substrates. In addition, sequence alignment and subsequent homology modelling of mouse liver 17beta-hydroxysteroid dehydrogenase and rat ovary 20alpha-hydroxysteroid dehydrogenase indicate that three loops on the C-terminal side of the barrel play potential roles in determining the positional and stereo-specificity of the hydroxysteroid dehydrogenases. Finally, we propose that the aldo-keto reductase superfamily may represent an example of divergent evolution from an ancestral multifunctional oxidoreductase and an example of convergent evolution to the same active-site constellation as the short

  10. Carbon-carbon double-bond reductases in nature.

    PubMed

    Huang, Minmin; Hu, Haihong; Ma, Li; Zhou, Quan; Yu, Lushan; Zeng, Su

    2014-08-01

    Reduction of C = C bonds by reductases, found in a variety of microorganisms (e.g. yeasts, bacteria, and lower fungi), animals, and plants has applications in the production of metabolites that include pharmacologically active drugs and other chemicals. Therefore, the reductase enzymes that mediate this transformation have become important therapeutic targets and biotechnological tools. These reductases are broad-spectrum, in that, they can act on isolation/conjugation C = C-bond compounds, α,β-unsaturated carbonyl compounds, carboxylic acids, acid derivatives, and nitro compounds. In addition, several mutations in the reductase gene have been identified, some associated with diseases. Several of these reductases have been cloned and/or purified, and studies to further characterize them and determine their structure in order to identify potential industrial biocatalysts are still in progress. In this study, crucial reductases for bioreduction of C = C bonds have been reviewed with emphasis on their principal substrates and effective inhibitors, their distribution, genetic polymorphisms, and implications in human disease and treatment.

  11. Distribution of Prx-linked hydroperoxide reductase activity among microorganisms.

    PubMed

    Takeda, Kouji; Nishiyama, Yoshitaka; Yoda, Koji; Watanabe, Toshihiro; Nimura-Matsune, Kaori; Mura, Kiyoshi; Tokue, Chiyoko; Katoh, Tetzuya; Kawasaki, Shinji; Niimura, Youichi

    2004-01-01

    Peroxiredoxin (Prx) constitutes a large family of enzymes found in microorganisms, animals, and plants, but the detection of the activities of Prx-linked hydroperoxide reductases (peroxiredoxin reductases) in cell extracts, and the purification based on peroxide reductase activity, have only been done in bacteria and Trypanosomatidae. A peroxiredoxin reductase (NADH oxidase) from a bacterium, Amphibacillus, displayed only poor activities in the presence of purified Prx from Saccharomyces or Synechocystis, while it is highly active in the presence of bacterial Prx. These results suggested that an enzyme system different from that in bacteria might exist for the reduction of Prx in yeast and cyanobacteria. Prx-linked hydroperoxide reductase activities were detected in cell extracts of Saccharomyces, Synechocystis, and Chlorella, and the enzyme activities of Saccharomyces and Chlorella were induced under vigorously aerated culture conditions and intensive light exposure conditions, respectively. Partial purification of Prx-linked peroxidase from the induced yeast cells indicated that the Prx-linked peroxidase system consists of two protein components, namely, thioredoxin and thioredoxin reductase. This finding is consistent with the previous report on its purification based on its protein protection activity against oxidation [Chae et al., J. Biol. Chem., 269, 27670-27678 (1994)]. In this study we have confirmed that Prx-linked peroxidase activity are widely distributed, not only in bacteria species and Trypanosomatidae, but also in yeast and photosynthetic microorganisms, and showed reconstitution of the activity from partially purified interspecies components.

  12. Engineering Styrene Monooxygenase for Biocatalysis: Reductase-Epoxidase Fusion Proteins.

    PubMed

    Heine, Thomas; Tucker, Kathryn; Okonkwo, Nonye; Assefa, Berhanegebriel; Conrad, Catleen; Scholtissek, Anika; Schlömann, Michael; Gassner, George; Tischler, Dirk

    2017-04-01

    The enantioselective epoxidation of styrene and related compounds by two-component styrene monooxygenases (SMOs) has targeted these enzymes for development as biocatalysts. In the present work, we prepare genetically engineered fusion proteins that join the C-terminus of the epoxidase (StyA) to the N-terminus of the reductase (StyB) through a linker peptide and demonstrate their utility as biocatalysts in the synthesis of Tyrain purple and other indigoid dyes. A single-vector expression system offers a simplified platform for transformation and expansion of the catalytic function of styrene monooxygenases, and the resulting fusion proteins are self-regulated and couple efficiently NADH oxidation to styrene epoxidation. We find that the reductase domain proceeds through a sequential ternary-complex mechanism at low FAD concentration and a double-displacement mechanism at higher concentrations of FAD. Single-turnover studies indicate an observed rate constant for FAD-to-FAD hydride transfer of ~8 s(-1). This step is rate limiting in the styrene epoxidation reaction and helps to ensure that flavin reduction and styrene epoxidation reactions proceed without wasteful side reactions. Comparison of the reductase activity of the fusion proteins with the naturally occurring reductase, SMOB, and N-terminally histidine-tagged reductase, NSMOB, suggests that the observed changes in catalytic mechanism are due in part to an increase in flavin-binding affinity associated with the N-terminal extension of the reductase.

  13. Molybdenum effector of fumarate reductase repression and nitrate reductase induction in Escherichia coli.

    PubMed Central

    Iuchi, S; Lin, E C

    1987-01-01

    In Escherichia coli the presence of nitrate prevents the utilization of fumarate as an anaerobic electron acceptor. The induction of the narC operon encoding the nitrate reductase is coupled to the repression of the frd operon encoding the fumarate reductase. This coupling is mediated by nitrate as an effector and the narL product as the regulatory protein (S. Iuchi and E. C. C. Lin, Proc. Natl. Acad. Sci. USA 84:3901-3905, 1987). The protein-ligand complex appears to control narC positively but frd negatively. In the present study we found that a molybdenum coeffector acted synergistically with nitrate in the regulation of frd and narC. In chlD mutants believed to be impaired in molybdate transport (or processing), full repression of phi(frd-lac) and full induction of phi(narC-lac) by nitrate did not occur unless the growth medium was directly supplemented with molybdate (1 microM). This requirement was not clearly manifested in wild-type cells, apparently because it was met by the trace quantities of molybdate present as a contaminant in the mineral medium. In chlB mutants, which are known to accumulate the Mo cofactor because of its failure to be inserted as a prosthetic group into proteins such as nitrate reductase, nitrate repression of frd and induction of narC were also intensified by molybdate supplementation. In this case a deficiency of the molybdenum coeffector might have resulted from enhanced feedback inhibition of molybdate transport (or processing) by the elevated level of the unutilized Mo cofactor. In addition, mutations in chlE, which are known to block the synthesis of the organic moiety of the Mo cofactor, lowered the threshold concentration of nitrate (< 1 micromole) necessary for frd repression and narC induction. These changes could be explained simply by the higher intracellular nitrate attainable in cells lacking the ability to destroy the effector. PMID:3301812

  14. Microsecond subdomain folding in dihydrofolate reductase.

    PubMed

    Arai, Munehito; Iwakura, Masahiro; Matthews, C Robert; Bilsel, Osman

    2011-07-08

    The characterization of microsecond dynamics in the folding of multisubdomain proteins has been a major challenge in understanding their often complex folding mechanisms. Using a continuous-flow mixing device coupled with fluorescence lifetime detection, we report the microsecond folding dynamics of dihydrofolate reductase (DHFR), a two-subdomain α/β/α sandwich protein known to begin folding in this time range. The global dimensions of early intermediates were monitored by Förster resonance energy transfer, and the dynamic properties of the local Trp environments were monitored by fluorescence lifetime detection. We found that substantial collapse occurs in both the locally connected adenosine binding subdomain and the discontinuous loop subdomain within 35 μs of initiation of folding from the urea unfolded state. During the fastest observable ∼550 μs phase, the discontinuous loop subdomain further contracts, concomitant with the burial of Trp residue(s), as both subdomains achieve a similar degree of compactness. Taken together with previous studies in the millisecond time range, a hierarchical assembly of DHFR--in which each subdomain independently folds, subsequently docks, and then anneals into the native conformation after an initial heterogeneous global collapse--emerges. The progressive acquisition of structure, beginning with a continuously connected subdomain and spreading to distal regions, shows that chain entropy is a significant organizing principle in the folding of multisubdomain proteins and single-domain proteins. Subdomain folding also provides a rationale for the complex kinetics often observed.

  15. Aldose reductase mediates retinal microglia activation

    SciTech Connect

    Chang, Kun-Che; Shieh, Biehuoy; Petrash, J. Mark

    2016-04-29

    Retinal microglia (RMG) are one of the major immune cells in charge of surveillance of inflammatory responses in the eye. In the absence of an inflammatory stimulus, RMG reside predominately in the ganglion layer and inner or outer plexiform layers. However, under stress RMG become activated and migrate into the inner nuclear layer (INL) or outer nuclear layer (ONL). Activated RMG in cell culture secrete pro-inflammatory cytokines in a manner sensitive to downregulation by aldose reductase inhibitors. In this study, we utilized CX3CR1{sup GFP} mice carrying AR mutant alleles to evaluate the role of AR on RMG activation and migration in vivo. When tested on an AR{sup WT} background, IP injection of LPS induced RMG activation and migration into the INL and ONL. However, this phenomenon was largely prevented by AR inhibitors or in AR null mice, or was exacerbated in transgenic mice that over-express AR. LPS-induced increases in ocular levels of TNF-α and CX3CL-1 in WT mice were substantially lower in AR null mice or were reduced by AR inhibitor treatment. These studies demonstrate that AR expression in RMG may contribute to the proinflammatory phenotypes common to various eye diseases such as uveitis and diabetic retinopathy. - Highlights: • AR inhibition prevents retinal microglial activation. • Endotoxin-induced ocular cytokine production is reduced in AR null mice. • Overexpression of AR spontaneously induces retinal microglial activation.

  16. Active sites of thioredoxin reductases: why selenoproteins?

    PubMed

    Gromer, Stephan; Johansson, Linda; Bauer, Holger; Arscott, L David; Rauch, Susanne; Ballou, David P; Williams, Charles H; Schirmer, R Heiner; Arnér, Elias S J

    2003-10-28

    Selenium, an essential trace element for mammals, is incorporated into a selected class of selenoproteins as selenocysteine. All known isoenzymes of mammalian thioredoxin (Trx) reductases (TrxRs) employ selenium in the C-terminal redox center -Gly-Cys-Sec-Gly-COOH for reduction of Trx and other substrates, whereas the corresponding sequence in Drosophila melanogaster TrxR is -Ser-Cys-Cys-Ser-COOH. Surprisingly, the catalytic competence of these orthologous enzymes is similar, whereas direct Sec-to-Cys substitution of mammalian TrxR, or other selenoenzymes, yields almost inactive enzyme. TrxRs are therefore ideal for studying the biology of selenocysteine by comparative enzymology. Here we show that the serine residues flanking the C-terminal Cys residues of Drosophila TrxRs are responsible for activating the cysteines to match the catalytic efficiency of a selenocysteine-cysteine pair as in mammalian TrxR, obviating the need for selenium. This finding suggests that the occurrence of selenoenzymes, which implies that the organism is selenium-dependent, is not necessarily associated with improved enzyme efficiency. Our data suggest that the selective advantage of selenoenzymes is a broader range of substrates and a broader range of microenvironmental conditions in which enzyme activity is possible.

  17. Functional properties and structural characterization of rice δ1-pyrroline-5-carboxylate reductase

    DOE PAGES

    Forlani, Giuseppe; Bertazzini, Michele; Zarattini, Marco; ...

    2015-07-28

    The majority of plant species accumulate high intracellular levels of proline to cope with hyperosmotic stress conditions. Proline synthesis from glutamate is tightly regulated at both the transcriptional and the translational levels, yet little is known about the mechanisms for post-translational regulation of the enzymatic activities involved. The gene coding in rice (Oryza sativa L.) for δ1-pyrroline-5-carboxylate (P5C) reductase, the enzyme that catalyzes the second and final step in this pathway, was isolated and expressed in Escherichia coli. The structural and functional properties of the affinity-purified protein were characterized. As for most species, rice P5C reductase was able to usemore » in vitro either NADH or NADPH as the electron donor. However, strikingly different effects of cations and anions were found depending on the pyridine nucleotide used, namely inhibition of NADH-dependent activity and stimulation of NADPH-dependent activity. Moreover, physiological concentrations of proline and NADP+ were strongly inhibitory for the NADH-dependent reaction, whereas the NADPH-dependent activity was mildly affected. Our results suggest that only NADPH may be used in vivo and that stress-dependent variations in ion homeostasis and NADPH/NADP+ ratio could modulate enzyme activity, being functional in promoting proline accumulation and potentially also adjusting NADPH consumption during the defense against hyperosmotic stress. The apparent molecular weight of the native protein observed in size exclusion chromatography indicated a high oligomerization state. We also report the first crystal structure of a plant P5C reductase at 3.40-Å resolution, showing a decameric quaternary assembly. It was possible to identify dynamic structural differences among rice, human, and bacterial enzymes.« less

  18. Prevention of toxic effects of mercuric chloride on Some male accessory organs in mice with a Multiherbal drug “Speman”

    PubMed Central

    Rathore, H.S.

    2000-01-01

    Adult Swiss albino male mice exposed to mercuric chloride via drinking water at 5 μg/ml for 100 days revealed significant reduction in the wet weight and severe histopathological changes in male accessory organs, poor level of serum testosterone and infertility. These effects were reduced remarkable and fertility was restored when drug (12.50 mg/mouse/day orally) was administered during mercury exposure for 100days or after Hg-exposure for next 60 days (Post therapy). Natural recovery after mercury exposure for 60 days remind ineffective. Probable action of herbal drug based on the presence of the active principles of constituents (i.e Orchis mascula, Mucuna pruriens, parmelia perlata, Argyreia speciosa, Tribulus terristris, Leptadenia reticulate, Lactuca scariola and Hygrophila spinosa) is discussed in detail. PMID:22556990

  19. Exposure to Silver Nanoparticles Inhibits Selenoprotein Synthesis and the Activity of Thioredoxin Reductase

    PubMed Central

    Srivastava, Milan; Singh, Sanjay

    2011-01-01

    Background: Silver nanoparticles (AgNPs) and silver (Ag)-based materials are increasingly being incorporated into consumer products, and although humans have been exposed to colloidal Ag in many forms for decades, this rise in the use of Ag materials has spurred interest into their toxicology. Recent reports have shown that exposure to AgNPs or Ag ions leads to oxidative stress, endoplasmic reticulum stress, and reduced cell proliferation. Previous studies have shown that Ag accumulates in tissues as silver sulfides (Ag2S) and silver selenide (Ag2Se). Objectives: In this study we investigated whether exposure of cells in culture to AgNPs or Ag ions at subtoxic doses would alter the effective metabolism of selenium, that is, the incorporation of selenium into selenoproteins. Methods: For these studies we used a keratinocyte cell model (HaCat) and a lung cell model (A549). We also tested (in vitro, both cellular and chemical) whether Ag ions could inhibit the activity of a key selenoenzyme, thioredoxin reductase (TrxR). Results: We found that exposure to AgNPs or far lower levels of Ag ions led to a dose-dependent inhibition of selenium metabolism in both cell models. The synthesis of protein was not altered under these conditions. Exposure to nanomolar levels of Ag ions effectively blocked selenium metabolism, suggesting that Ag ion leaching was likely the mechanism underlying observed changes during AgNP exposure. Exposure likewise inhibited TrxR activity in cultured cells, and Ag ions were potent inhibitors of purified rat TrxR isoform 1 (cytosolic) (TrxR1) enzyme. Conclusions: Exposure to AgNPs leads to the inhibition of selenoprotein synthesis and inhibition of TrxR1. Further, we propose these two sites of action comprise the likely mechanism underlying increases in oxidative stress, increases endoplasmic reticulum stress, and reduced cell proliferation during exposure to Ag. PMID:21965219

  20. Mercuric chloride poisoning

    MedlinePlus

    ... help if this information is not immediately available. Poison Control Your local poison center can be reached directly by calling the national toll-free Poison Help hotline (1-800-222-1222) from anywhere ...

  1. Mercuric oxide poisoning

    MedlinePlus

    ... help if this information is not immediately available. Poison Control Your local poison center can be reached directly by calling the national toll-free Poison Help hotline (1-800-222-1222) from anywhere ...

  2. Reaction of methanol with chlorate ions in acid solution containing Hg{sup +2} by NMR

    SciTech Connect

    Ernst, W.R.; Indu, B.; Crump, B.; Gelbaum, L.T.

    1996-05-01

    The reaction rate of methanol was measured in solutions of sodium chlorate and sulfuric acid at several levels of temperature and concentration, in the presence of mercuric nitrate. The progress of the reaction was monitored by proton NMR signals corresponding to methanol and formic acid. Chlorine dioxide formation was suppressed by adding mercuric nitrate, which was shown earlier to catalyze the disproportionation of the intermediate species, chlorous acid, and sequester chloride ions. The reaction is first order in methanol and chlorate concentration and in the Hammett acidity function. The reaction of formic acid, sodium chlorate and sulfuric acid was also studied using the same technique. Formic acid was stable and did not react with chlorate at a measurable rate, even at concentrations and temperatures of a commercial process. This study related to commercial processes that produce chlorine dioxide by reducing chlorate ions with methanol. Chlorine dioxide is an oxidizing chemical that is used in water purification and is replacing chlorine in many chemical bleaching processes because of environmental concerns.

  3. Sulfite reductase protects plants against sulfite toxicity.

    PubMed

    Yarmolinsky, Dmitry; Brychkova, Galina; Fluhr, Robert; Sagi, Moshe

    2013-02-01

    Plant sulfite reductase (SiR; Enzyme Commission 1.8.7.1) catalyzes the reduction of sulfite to sulfide in the reductive sulfate assimilation pathway. Comparison of SiR expression in tomato (Solanum lycopersicum 'Rheinlands Ruhm') and Arabidopsis (Arabidopsis thaliana) plants revealed that SiR is expressed in a different tissue-dependent manner that likely reflects dissimilarity in sulfur metabolism between the plant species. Using Arabidopsis and tomato SiR mutants with modified SiR expression, we show here that resistance to ectopically applied sulfur dioxide/sulfite is a function of SiR expression levels and that plants with reduced SiR expression exhibit higher sensitivity than the wild type, as manifested in pronounced leaf necrosis and chlorophyll bleaching. The sulfite-sensitive mutants accumulate applied sulfite and show a decline in glutathione levels. In contrast, mutants that overexpress SiR are more tolerant to sulfite toxicity, exhibiting little or no damage. Resistance to high sulfite application is manifested by fast sulfite disappearance and an increase in glutathione levels. The notion that SiR plays a role in the protection of plants against sulfite is supported by the rapid up-regulation of SiR transcript and activity within 30 min of sulfite injection into Arabidopsis and tomato leaves. Peroxisomal sulfite oxidase transcripts and activity levels are likewise promoted by sulfite application as compared with water injection controls. These results indicate that, in addition to participating in the sulfate assimilation reductive pathway, SiR also plays a role in protecting leaves against the toxicity of sulfite accumulation.

  4. Molecular dissection of a putative iron reductase from Desulfotomaculum reducens MI-1

    SciTech Connect

    Li, Zhi; Kim, David D.; Nelson, Ornella D.; Otwell, Annie E.; Richardson, Ruth E.; Callister, Stephen J.; Lin, Hening

    2015-10-08

    Desulfotomaculum reducens MI-1 is a Firmicute strain capable of reducing a variety of heavy metal ions and has a great potential in heavy metal bioremediation.We recently identified Dred_2421 as a potential iron reductase through proteomic study of D. reducens. The current study examines its iron-reduction mechanism. Dred_2421, like its close homolog from Escherichia coli (2, 4-dienoyl-CoA reductase), has an FMN-binding N-terminal domain (NTD), an FAD-binding C-terminal domain (CTD), and a 4Fee4S cluster between the two domains. To understand the mechanism of the iron-reduction activity and the role of each domain, we generated a series of variants for each domain and investigated their iron reduction activity. Our results suggest that CTD is the main contributor of the iron-reduction activity, and that NTD and the 4Fee4S cluster are not directly involved in such activity. This study provides a mechanistic understanding of the ironereductase activity of Dred_2421 and may also help to elucidate other physiological activities this enzyme may have.

  5. Structural Insight into Dihydrodipicolinate Reductase from Corybebacterium glutamicum for Lysine Biosynthesis.

    PubMed

    Sagong, Hye-Young; Kim, Kyung-Jin

    2016-02-01

    Dihydrodipicolinate reductase is an enzyme that converts dihydrodipicolinate to tetrahydrodipicolinate using an NAD(P)H cofactor in L-lysine biosynthesis. To increase the understanding of the molecular mechanisms of lysine biosynthesis, we determined the crystal structure of dihydrodipicolinate reductase from Corynebacterium glutamicum (CgDapB). CgDapB functions as a tetramer, and each protomer is composed of two domains, an Nterminal domain and a C-terminal domain. The N-terminal domain mainly contributes to nucleotide binding, whereas the C-terminal domain is involved in substrate binding. We elucidated the mode of cofactor binding to CgDapB by determining the crystal structure of the enzyme in complex with NADP(+) and found that CgDapB utilizes both NADH and NADPH as cofactors. Moreover, we determined the substrate binding mode of the enzyme based on the coordination mode of two sulfate ions in our structure. Compared with Mycobacterium tuberculosis DapB in complex with its cofactor and inhibitor, we propose that the domain movement for active site constitution occurs when both cofactor and substrate bind to the enzyme.

  6. Purification and properties of proline reductase from Clostridium sticklandii.

    PubMed

    Seto, B; Stadtman, T C

    1976-04-25

    Proline reductase of Clostridium sticklandii is a membrane-bound protein and is released by treatment with detergents. The enzyme has been purified to homogeneity and is estimated by gel filtration and sedimentation equilibrium centrifugation to have a molecular weight of 298,000 to 327,000. A minimum molecular weight of 30,000 to 31,000 was calculated on the basis of sodium dodecyl sulfate-acrylamide gel electrophoresis and amino acid composition. Amino acid analysis showed a preponderance of acidic amino acids. No tryptophan was detected in the protein either spectrophotometrically or by amino acid analysis. A total of 20 sulfhydryl groups measured by titration of the reduced protein with 5,5'-dithiobis(2-nitrobenzoic acid) is in agreement with 20 cystic acid residues determined in hydrolysates of performic acid-oxidized protein. No molybdenum, iron, or selenium was found in the pure protein. Although NADH is the physiological electron donor for the proline reductase complex, the purified 300,000 molecular weight reductase component is inactive in the presence of NADH in vitro. Dithiothreitol, in contrast, can serve as electron donor both for unpurified (putative proline reductase complex) and purified proline reductase in vitro.

  7. Potential use of aldose reductase inhibitors to prevent diabetic complications.

    PubMed

    Zenon, G J; Abobo, C V; Carter, B L; Ball, D W

    1990-06-01

    Reviewed are (1) the biochemical basis and pathophysiology of diabetic complications and (2) the structure-activity relationships, pharmacology, pharmacokinetics, clinical trials, and adverse effects of aldose reductase inhibitors (ARIs). ARIs are a new class of drugs potentially useful in preventing diabetic complications, the most widely studied of which have been cataracts and neuropathy. ARIs inhibit aldose reductase, the first, rate-limiting enzyme in the polyol metabolic pathway. In nonphysiological hyperglycemia the activity of hexokinase becomes saturated while that of aldose reductase is enhanced, resulting in intracellular accumulation of sorbitol. Because sorbitol does not readily penetrate the cell membrane it can persist within cells, which may lead to diabetic complications. ARIs are a class of structurally dissimilar compounds that include carboxylic acid derivatives, flavonoids, and spirohydantoins. The major pharmacologic action of an ARI involves competitive binding to aldose reductase and consequent blocking of sorbitol production. ARIs delay cataract formation in animals, but the role of aldose reductase in cataract formation in human diabetics has not been established. The adverse effects of ARIs include hypersensitivity reactions. Although the polyol pathway may not be solely responsible for diabetic complications, studies suggest that therapy with ARIs could be beneficial. Further research is needed to determine the long-term impact and adverse effects of ARIs in the treatment of diabetic complications.

  8. Uterine glutathione reductase activity: modulation by estrogens and progesterone.

    PubMed

    Díaz-Flores, M; Baiza-Gutman, L A; Pedrón, N N; Hicks, J J

    1999-10-29

    The aim of this study was to determine whether glutathione reductase activity in uterine tissue is regulated by sex hormones. In spayed rats uterine glutathione reductase was significantly increased by exogenous estrogen (P< 0.01), progesterone (P< 0.01) or estrogen plus progesterone (P<0.01). When enzyme activity is expressed per mg protein, daily administration of estrogen or progesterone induces a progressive increase of this enzyme between 24 to 48 h or 24 to 72 h of treatment, respectively. Whereas the combination of both steroids causes an earlier and higher increase in glutathione reductase activity at 24 h of treatment. Estradiol singly or in combination with progesterone induced the highest protein concentration in the uterus. Whereas uterine DNA concentration is only significantly affected by estradiol. Our results suggest that uterine glutathione reductase is regulated by estradiol and progesterone and may be involved in maintaining levels of reduced glutathione in the uterus. This compound may be required for control of the redox state of thiol groups and in detoxification reactions involving H2O2 and electrophylic substances. The antioxidant action of estrogens is partially due to the stimulation of glutathione reductase.

  9. Bacterial morphinone reductase is related to Old Yellow Enzyme.

    PubMed Central

    French, C E; Bruce, N C

    1995-01-01

    Morphinone reductase, produced by Pseudomonas putida M10, catalyses the NADH-dependent saturation of the carbon-carbon double bond of morphinone and codeinone, and is believed to be involved in the metabolism of morphine and codeine. The structural gene encoding morphinone reductase, designated morB, was cloned from Ps. putida M10 genomic DNA by the use of degenerate oligonucleotide probes based on elements of the amino acid sequence of the purified enzyme. Sequence analysis and structural characteristics indicated that morphinone reductase is related to the flavoprotein alpha/beta-barrel oxidoreductases, and is particularly similar to Old Yellow Enzyme of Saccharomyces spp. and the related oestrogen-binding protein of Candida albicans. Expressed sequence tags from several plant species show high homology to these enzymes, suggesting the presence of a family of enzymes conserved in plants and fungi. Although related bacterial proteins are known, morphinone reductase appears to be more similar to the eukaryotic proteins. Morphinone reductase was overexpressed in Escherichia coli, and has potential applications for the industrial preparation of semisynthetic opiates. Images Figure 1 Figure 5 PMID:8554504

  10. HMG-CoA reductase guides migrating primordial germ cells.

    PubMed

    Van Doren, M; Broihier, H T; Moore, L A; Lehmann, R

    1998-12-03

    The enzyme 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase is best known for catalysing a rate-limiting step in cholesterol biosynthesis, but it also participates in the production of a wide variety of other compounds. Some clinical benefits attributed to inhibitors of HMG-CoA reductase are now thought to be independent of any serum cholesterol-lowering effect. Here we describe a new cholesterol-independent role for HMG-CoA reductase, in regulating a developmental process: primordial germ cell migration. We show that in Drosophila this enzyme is highly expressed in the somatic gonad and that it is necessary for primordial germ cells to migrate to this tissue. Misexpression of HMG-CoA reductase is sufficient to attract primordial germ cells to tissues other than the gonadal mesoderm. We conclude that the regulated expression of HMG-CoA reductase has a critical developmental function in providing spatial information to guide migrating primordial germ cells.

  11. Dynamic Control of Electron Transfers in Diflavin Reductases

    PubMed Central

    Aigrain, Louise; Fatemi, Fataneh; Frances, Oriane; Lescop, Ewen; Truan, Gilles

    2012-01-01

    Diflavin reductases are essential proteins capable of splitting the two-electron flux from reduced pyridine nucleotides to a variety of one electron acceptors. The primary sequence of diflavin reductases shows a conserved domain organization harboring two catalytic domains bound to the FAD and FMN flavins sandwiched by one or several non-catalytic domains. The catalytic domains are analogous to existing globular proteins: the FMN domain is analogous to flavodoxins while the FAD domain resembles ferredoxin reductases. The first structural determination of one member of the diflavin reductases family raised some questions about the architecture of the enzyme during catalysis: both FMN and FAD were in perfect position for interflavin transfers but the steric hindrance of the FAD domain rapidly prompted more complex hypotheses on the possible mechanisms for the electron transfer from FMN to external acceptors. Hypotheses of domain reorganization during catalysis in the context of the different members of this family were given by many groups during the past twenty years. This review will address the recent advances in various structural approaches that have highlighted specific dynamic features of diflavin reductases. PMID:23203109

  12. [Methylenetetrahydrofolate reductase and methionine synthase reductase gene polymorphisms in ethnic Han women from Linyi].

    PubMed

    Zhang, Yan-li; Lu, Yan-qiang; Li, Hua-feng; Rui, Xin-yi; Zhang, Li-jun; Wu, Chuan-ye; Fang, Ai-min; Wang, Gui-xi

    2012-12-01

    To explore the distribution of genetic polymorphisms of methylenetetrahydrofolate reductase (MTHFR) 677C/T, 1298A/C and methionine synthase reductase (MTRR) 66A/G among ethnic Han females from Linyi, and to correlate it with serum level of homocysteine (Hcy). A cross-sectional study was carried out. Oral epithelial cell samples were collected from 825 subjects. MTHFR and MTRR gene polymorphisms were determined with a Taqman-Minor Groove Binder (MGB) method. Distribution of gene polymorphisms was analyzed and compared with others regions of China including Weifang, Zhengzhou, Deyang and Hainan. A biochemical assay was also carried out to determine the total Hcy in plasma of 281 subjects. The reductase activity of MTHFR was classified into decreased and stable groups according to genetic polymorphism of MTHFR. Correlation between MTHFR groups and total Hcy level were also explored. (1) The frequencies of MTHFR677CC, CT and TT genotypes of the selected subjects were 16.7%, 48.3% and 35.0%, respectively. The frequencies of MTHFR 1298AA, AC and CC genotypes were 76.0%, 21.6% and 2.4%, respectively. And those of MTRR 66AA, AG and GG genotypes were 54.7%, 39.4% and 5.9%, respectively. For the selected subjects, their frequency of MTHFR 677TT genotype was higher than that of Deyang and Hainan (P< 0.01), whilst the frequency of MTHFR 1298CC genotype was lower than that of Deyang and Hainan (P < 0.01), and the frequency of MTRR 66 GG genotype was lower than that of Hainan (P< 0.01). (2) The Hcy level for those with decreased MTHFR activity was significantly higher than those with stable MTHFR activity (P< 0.05). MTHFR gene 677C/T, 1298A/C and MTRR 66A/G polymorphisms in ethnic Han women from Linyi have differed significantly from other regions of China. Decreased MTHFR activity caused by genetic polymorphisms is a risk factor for raised Hcy level.

  13. [Ligand spectrum of hemoglobin activity of methemoglobin-reductase and hemolytic resistance of erythrocytes during chronic exposure to nitrates].

    PubMed

    Kiiza, D A; Artiukh, V P; Starodub, N F; Khmel'nitskiĭ, G A

    1992-01-01

    It is found that nitrite-ions formed as a result of biotransformation during long term feeding of calves with sodium and potassium nitrates induce changes in some biochemical parameters of blood, including HS-glutathione content in erythrocytes, acid hemolytic resistance of erythrocytes, activity of NAD-dependent methemoglobin-reductase, correlation of ligand forms of hemoglobin and its total content. It is supposed that the observed changes are of an adaptational character and, as a whole, provide for the optimization of both quantitative and qualitative composition of population of erythroid cells at the expense of erythropoiesis intensification.

  14. Proanthocyanidin synthesis in Theobroma cacao: genes encoding anthocyanidin synthase, anthocyanidin reductase, and leucoanthocyanidin reductase

    PubMed Central

    2013-01-01

    Background The proanthocyanidins (PAs), a subgroup of flavonoids, accumulate to levels of approximately 10% total dry weight of cacao seeds. PAs have been associated with human health benefits and also play important roles in pest and disease defense throughout the plant. Results To dissect the genetic basis of PA biosynthetic pathway in cacao (Theobroma cacao), we have isolated three genes encoding key PA synthesis enzymes, anthocyanidin synthase (ANS), anthocyanidin reductase (ANR) and leucoanthocyanidin reductase (LAR). We measured the expression levels of TcANR, TcANS and TcLAR and PA content in cacao leaves, flowers, pod exocarp and seeds. In all tissues examined, all three genes were abundantly expressed and well correlated with PA accumulation levels, suggesting their active roles in PA synthesis. Overexpression of TcANR in an Arabidopsis ban mutant complemented the PA deficient phenotype in seeds and resulted in reduced anthocyanidin levels in hypocotyls. Overexpression of TcANS in tobacco resulted in increased content of both anthocyanidins and PAs in flower petals. Overexpression of TcANS in an Arabidopsis ldox mutant complemented its PA deficient phenotype in seeds. Recombinant TcLAR protein converted leucoanthocyanidin to catechin in vitro. Transgenic tobacco overexpressing TcLAR had decreased amounts of anthocyanidins and increased PAs. Overexpressing TcLAR in Arabidopsis ldox mutant also resulted in elevated synthesis of not only catechin but also epicatechin. Conclusion Our results confirm the in vivo function of cacao ANS and ANR predicted based on sequence homology to previously characterized enzymes from other species. In addition, our results provide a clear functional analysis of a LAR gene in vivo. PMID:24308601

  15. Proanthocyanidin synthesis in Theobroma cacao: genes encoding anthocyanidin synthase, anthocyanidin reductase, and leucoanthocyanidin reductase.

    PubMed

    Liu, Yi; Shi, Zi; Maximova, Siela; Payne, Mark J; Guiltinan, Mark J

    2013-12-05

    The proanthocyanidins (PAs), a subgroup of flavonoids, accumulate to levels of approximately 10% total dry weight of cacao seeds. PAs have been associated with human health benefits and also play important roles in pest and disease defense throughout the plant. To dissect the genetic basis of PA biosynthetic pathway in cacao (Theobroma cacao), we have isolated three genes encoding key PA synthesis enzymes, anthocyanidin synthase (ANS), anthocyanidin reductase (ANR) and leucoanthocyanidin reductase (LAR). We measured the expression levels of TcANR, TcANS and TcLAR and PA content in cacao leaves, flowers, pod exocarp and seeds. In all tissues examined, all three genes were abundantly expressed and well correlated with PA accumulation levels, suggesting their active roles in PA synthesis. Overexpression of TcANR in an Arabidopsis ban mutant complemented the PA deficient phenotype in seeds and resulted in reduced anthocyanidin levels in hypocotyls. Overexpression of TcANS in tobacco resulted in increased content of both anthocyanidins and PAs in flower petals. Overexpression of TcANS in an Arabidopsis ldox mutant complemented its PA deficient phenotype in seeds. Recombinant TcLAR protein converted leucoanthocyanidin to catechin in vitro. Transgenic tobacco overexpressing TcLAR had decreased amounts of anthocyanidins and increased PAs. Overexpressing TcLAR in Arabidopsis ldox mutant also resulted in elevated synthesis of not only catechin but also epicatechin. Our results confirm the in vivo function of cacao ANS and ANR predicted based on sequence homology to previously characterized enzymes from other species. In addition, our results provide a clear functional analysis of a LAR gene in vivo.

  16. Sulfur Isotope Effects of Dissimilatory Sulfite Reductase.

    PubMed

    Leavitt, William D; Bradley, Alexander S; Santos, André A; Pereira, Inês A C; Johnston, David T

    2015-01-01

    The precise interpretation of environmental sulfur isotope records requires a quantitative understanding of the biochemical controls on sulfur isotope fractionation by the principle isotope-fractionating process within the S cycle, microbial sulfate reduction (MSR). Here we provide the only direct observation of the major ((34)S/(32)S) and minor ((33)S/(32)S, (36)S/(32)S) sulfur isotope fractionations imparted by a central enzyme in the energy metabolism of sulfate reducers, dissimilatory sulfite reductase (DsrAB). Results from in vitro sulfite reduction experiments allow us to calculate the in vitro DsrAB isotope effect in (34)S/(32)S (hereafter, [Formula: see text]) to be 15.3 ± 2‰, 2σ. The accompanying minor isotope effect in (33)S, described as [Formula: see text], is calculated to be 0.5150 ± 0.0012, 2σ. These observations facilitate a rigorous evaluation of the isotopic fractionation associated with the dissimilatory MSR pathway, as well as of the environmental variables that govern the overall magnitude of fractionation by natural communities of sulfate reducers. The isotope effect induced by DsrAB upon sulfite reduction is a factor of 0.3-0.6 times prior indirect estimates, which have ranged from 25 to 53‰ in (34)εDsrAB. The minor isotope fractionation observed from DsrAB is consistent with a kinetic or equilibrium effect. Our in vitro constraints on the magnitude of [Formula: see text] is similar to the median value of experimental observations compiled from all known published work, where (34)ε r-p = 16.1‰ (r-p indicates reactant vs. product, n = 648). This value closely matches those of MSR operating at high sulfate reduction rates in both laboratory chemostat experiments ([Formula: see text] 17.3 ± 1.5‰, 2σ) and in modern marine sediments ([Formula: see text] 17.3 ± 3.8‰). Targeting the direct isotopic consequences of a specific enzymatic processes is a fundamental step toward a biochemical foundation for reinterpreting the

  17. Selenium in thioredoxin reductase: a mechanistic perspective.

    PubMed

    Lacey, Brian M; Eckenroth, Brian E; Flemer, Stevenson; Hondal, Robert J

    2008-12-02

    Most high M(r) thioredoxin reductases (TRs) have the unusual feature of utilizing a vicinal disulfide bond (Cys(1)-Cys(2)) which forms an eight-membered ring during the catalytic cycle. Many eukaryotic TRs have replaced the Cys(2) position of the dyad with the rare amino acid selenocysteine (Sec). Here we demonstrate that Cys- and Sec-containing TRs are distinguished by the importance each class of enzymes places on the eight-membered ring structure in the catalytic cycle. This hypothesis was explored by studying the truncated enzyme missing the C-terminal ring structure in conjunction with oxidized peptide substrates to investigate the reduction and opening of this dyad. The peptide substrates were identical in sequence to the missing part of the enzyme, containing either a disulfide or selenylsulfide linkage, but were differentiated by the presence (cyclic) and absence (acyclic) of the ring structure. The ratio of these turnover rates informs that the ring is only of modest importance for the truncated mouse mitochondrial Sec-TR (ring/no ring = 32), while the ring structure is highly important for the truncated Cys-TRs from Drosophila melanogaster and Caenorhabditis elegans (ring/no ring > 1000). All three enzymes exhibit a similar dependence upon leaving group pK(a) as shown by the use of the acyclic peptides as substrates. These two factors can be reconciled for Cys-TRs if the ring functions to simultaneously allow for attack by a nearby thiolate while correctly positioning the leaving group sulfur atom to accept a proton from the enzymic general acid. For Sec-TRs the ring is unimportant because the lower pK(a) of the selenol relative to a thiol obviates its need to be protonated upon S-Se bond scission and permits physical separation of the selenol and the general acid. Further study of the biochemical properties of the truncated Cys and Sec TR enzymes demonstrates that the chemical advantage conferred on the eukaryotic enzyme by a selenol is the ability to

  18. Sulfur Isotope Effects of Dissimilatory Sulfite Reductase

    PubMed Central

    Leavitt, William D.; Bradley, Alexander S.; Santos, André A.; Pereira, Inês A. C.; Johnston, David T.

    2015-01-01

    The precise interpretation of environmental sulfur isotope records requires a quantitative understanding of the biochemical controls on sulfur isotope fractionation by the principle isotope-fractionating process within the S cycle, microbial sulfate reduction (MSR). Here we provide the only direct observation of the major (34S/32S) and minor (33S/32S, 36S/32S) sulfur isotope fractionations imparted by a central enzyme in the energy metabolism of sulfate reducers, dissimilatory sulfite reductase (DsrAB). Results from in vitro sulfite reduction experiments allow us to calculate the in vitro DsrAB isotope effect in 34S/32S (hereafter, 34εDsrAB) to be 15.3 ± 2‰, 2σ. The accompanying minor isotope effect in 33S, described as 33λDsrAB, is calculated to be 0.5150 ± 0.0012, 2σ. These observations facilitate a rigorous evaluation of the isotopic fractionation associated with the dissimilatory MSR pathway, as well as of the environmental variables that govern the overall magnitude of fractionation by natural communities of sulfate reducers. The isotope effect induced by DsrAB upon sulfite reduction is a factor of 0.3–0.6 times prior indirect estimates, which have ranged from 25 to 53‰ in 34εDsrAB. The minor isotope fractionation observed from DsrAB is consistent with a kinetic or equilibrium effect. Our in vitro constraints on the magnitude of 34εDsrAB is similar to the median value of experimental observations compiled from all known published work, where 34εr−p = 16.1‰ (r–p indicates reactant vs. product, n = 648). This value closely matches those of MSR operating at high sulfate reduction rates in both laboratory chemostat experiments (34εSO4−H2S =  17.3 ± 1.5‰, 2σ) and in modern marine sediments (34εSO4−H2S =  17.3 ± 3.8‰). Targeting the direct isotopic consequences of a specific enzymatic processes is a fundamental step toward a biochemical foundation for reinterpreting the biogeochemical and geobiological sulfur isotope records in

  19. An (R)-Imine Reductase Biocatalyst for the Asymmetric Reduction of Cyclic Imines.

    PubMed

    Hussain, Shahed; Leipold, Friedemann; Man, Henry; Wells, Elizabeth; France, Scott P; Mulholland, Keith R; Grogan, Gideon; Turner, Nicholas J

    2015-02-01

    Although the range of biocatalysts available for the synthesis of enantiomerically pure chiral amines continues to expand, few existing methods provide access to secondary amines. To address this shortcoming, we have over-expressed the gene for an (R)-imine reductase [(R)-IRED] from Streptomyces sp. GF3587 in Escherichia coli to create a recombinant whole-cell biocatalyst for the asymmetric reduction of prochiral imines. The (R)-IRED was screened against a panel of cyclic imines and two iminium ions and was shown to possess high catalytic activity and enantioselectivity. Preparative-scale synthesis of the alkaloid (R)-coniine (90 % yield; 99 % ee) from the imine precursor was performed on a gram-scale. A homology model of the enzyme active site, based on the structure of a closely related (R)-IRED from Streptomyces kanamyceticus, was constructed and used to identify potential amino acids as targets for mutagenesis.

  20. Hexavalent Chromate Reductase Activity in Cell Free Extracts of Penicillium sp.

    PubMed Central

    Arévalo-Rangel, Damaris L.; Cárdenas-González, Juan F.; Martínez-Juárez, Víctor M.; Acosta-Rodríguez, Ismael

    2013-01-01

    A chromium-resistant fungus isolated from contaminated air with industrial vapors can be used for reducing toxic Cr(VI) to Cr(III). This study analyzes in vitro reduction of hexavalent chromium using cell free extract(s) of the fungus that was characterized based on optimal temperature, pH, use of electron donors, metal ions and initial Cr(VI) concentration in the reaction mixture. This showed the highest activity at 37°C and pH 7.0; there is an increase in Cr(VI) reductase activity with addition of NADH as an electron donor, and it was highly inhibited by Hg2+, Ca2+ and Mg2+, and azide, EDTA, and KCN. PMID:24027493

  1. Evidence for the involvement of two heterodisulfide reductases in the energy-conserving system of Methanomassiliicoccus luminyensis.

    PubMed

    Kröninger, Lena; Berger, Stefanie; Welte, Cornelia; Deppenmeier, Uwe

    2016-02-01

    Methanomassiliicoccus luminyensis was isolated from the human gut, and requires H2 and methanol or methylamines to produce methane. The organism lacks cytochromes, indicating that it cannot couple membrane-bound electron transfer reactions with extrusion of H(+) or Na(+) ions using known methanogenic pathways. Furthermore, M. luminyensis contains a soluble hydrogenase/heterodisulfide reductase complex (MvhAGD/HdrABC) as found in obligate hydrogenotrophic methanogens, but the energy-conserving methyltransferase (MtrA-H) is absent. Thus, the question arises as to how this species synthesizes ATP. We present evidence that M. luminyensis uses two types of heterodisulfide reductases (HdrABC and HdrD) in a novel process for energy conservation. Quantitative RT-PCR studies revealed that genes encoding these heterodisulfide reductases showed high expression levels. Other genes with high transcript abundance were fpoA (part of the operon encoding the 'headless' F420 H2 dehydrogenase) and atpB (part of the operon encoding the A1 Ao ATP synthase). High activities of the soluble heterodisulfide reductase HdrABC and the hydrogenase MvhADG were found in the cytoplasm of M. luminyensis. Also, heterologously produced HdrD was able to reduce CoM-S-S-CoB using reduced methylviologen as an electron donor. We propose that membrane-bound electron transfer is based on conversion of two molecules of methanol and concurrent formation of two molecules of the heterodisulfide CoM-S-S-CoB. First the HdrABC/MvhADG complex catalyzes the H2 -dependent reduction of CoM-S-S-CoB and formation of reduced ferredoxin. In a second cycle, reduced ferredoxin is oxidized by the 'headless' F420 H2 dehydrogenase, thereby translocating up to 4 H(+) across the membrane, and electrons are channeled to HdrD for reduction of the second heterodisulfide. © 2015 FEBS.

  2. A stable mercury-containing complex of the organomercurial lyase MerB: catalysis, product release, and direct transfer to MerA.

    PubMed

    Benison, Gregory C; Di Lello, Paola; Shokes, Jacob E; Cosper, Nathaniel J; Scott, Robert A; Legault, Pascale; Omichinski, James G

    2004-07-06

    Bacteria isolated from organic mercury-contaminated sites have developed a system of two enzymes that allows them to efficiently convert both ionic and organic mercury compounds to the less toxic elemental mercury. Both enzymes are encoded on the mer operon and require sulfhydryl-bound substrates. The first enzyme is an organomercurial lyase (MerB), and the second enzyme is a mercuric ion reductase (MerA). MerB catalyzes the protonolysis of the carbon-mercury bond, resulting in the formation of a reduced carbon compound and inorganic ionic mercury. Of several mercury-containing MerB complexes that we attempted to prepare, the most stable was a complex consisting of the organomercurial lyase (MerB), a mercuric ion, and a molecule of the MerB inhibitor dithiothreitol (DTT). Nuclear magnetic resonance (NMR) spectroscopy and extended X-ray absorption fine structure spectroscopy of the MerB/Hg/DTT complex have shown that the ligands to the mercuric ion in the complex consist of both sulfurs from the DTT molecule and one cysteine ligand, C96, from the protein. The stability of the MerB/Hg/DTT complex, even in the presence of a large excess of competing cysteine, has been demonstrated by NMR and dialysis. We used an enzyme buffering test to determine that the MerB/Hg/DTT complex acts as a substrate for the mercuric reductase MerA. The observed MerA activity is higher than the expected activity assuming free diffusion of the mercuric ion from MerB to MerA. This suggests that the mercuric ion can be transferred between the two enzymes by a direct transfer mechanism.

  3. Do cytochromes function as oxygen sensors in the regulation of nitrate reductase biosynthesis?

    PubMed Central

    MacGregor, C H; Bishop, C W

    1977-01-01

    The observation that oxygen represses nitrate reductase biosynthesis in a hemA mutant grown aerobically with or without delta-aminolevulinic acid indicates that cytochromes are not responsible for nitrate reductase repression in aerobically grown cells. PMID:326768

  4. Studies on the regulation of assimilatory nitrate reductase in Ankistrodesmus braunii.

    PubMed

    Diez, J; Chaparro, A; Vega, J M; Relimpio, A

    1977-01-01

    In the green alga Ankistrodesmus braunii, all the activities associated with the nitrate reductase complex (i.e., NAD(P)H-nitrate reductase, NAD(P)H-cytochrome c reductase and FMNH2-or MVH-nitrate reductase) are nutritionally repressed by ammonia or methylamine. Besides, ammonia or methylamine promote in vivo the reversible inactivation of nitrate reductase, but not of NAD(P)H-cytochrome c reductase. Subsequent removal of the inactivating agent from the medium causes reactivation of the inactive enzyme. Menadione has a striking stimulation on the in vivo reactivation of the inactive enzyme. The nitrate reductase activities, but not the diaphorase activity, can be inactivated in vitro by preincubating a partially purified enzyme preparation with NADH or NADPH. ADP, in the presence of Mg(2+), presents a cooperative effect with NADH in the in vitro inactivation of nitrate reductase. This effect appears to be maximum at a concentration of ADP equimolecular with that of NADH.

  5. Role of Conserved Glycine in Zinc-dependent Medium Chain Dehydrogenase/Reductase Superfamily*

    PubMed Central

    Tiwari, Manish Kumar; Singh, Raushan Kumar; Singh, Ranjitha; Jeya, Marimuthu; Zhao, Huimin; Lee, Jung-Kul

    2012-01-01

    The medium-chain dehydrogenase/reductase (MDR) superfamily consists of a large group of enzymes with a broad range of activities. Members of this superfamily are currently the subject of intensive investigation, but many aspects, including the zinc dependence of MDR superfamily proteins, have not yet have been adequately investigated. Using a density functional theory-based screening strategy, we have identified a strictly conserved glycine residue (Gly) in the zinc-dependent MDR superfamily. To elucidate the role of this conserved Gly in MDR, we carried out a comprehensive structural, functional, and computational analysis of four MDR enzymes through a series of studies including site-directed mutagenesis, isothermal titration calorimetry, electron paramagnetic resonance (EPR), quantum mechanics, and molecular mechanics analysis. Gly substitution by other amino acids posed a significant threat to the metal binding affinity and activity of MDR superfamily enzymes. Mutagenesis at the conserved Gly resulted in alterations in the coordination of the catalytic zinc ion, with concomitant changes in metal-ligand bond length, bond angle, and the affinity (Kd) toward the zinc ion. The Gly mutants also showed different spectroscopic properties in EPR compared with those of the wild type, indicating that the binding geometries of the zinc to the zinc binding ligands were changed by the mutation. The present results demonstrate that the conserved Gly in the GHE motif plays a role in maintaining the metal binding affinity and the electronic state of the catalytic zinc ion during catalysis of the MDR superfamily enzymes. PMID:22500022

  6. S-Mercuration of ubiquitin carboxyl-terminal hydrolase L1 through Cys152 by methylmercury causes inhibition of its catalytic activity and reduction of monoubiquitin levels in SH-SY5Y cells.

    PubMed

    Toyama, Takashi; Abiko, Yumi; Katayama, Yuko; Kaji, Toshiyuki; Kumagai, Yoshito

    2015-12-01

    Methylmercury (MeHg) is an environmental electrophile that covalently modifies cellular proteins. In this study, we identified proteins that undergo S-mercuration by MeHg. By combining two-dimensional SDS-PAGE, atomic absorption spectrometry and ultra performance liquid chromatography mass spectrometry (UPLC/MS/MS), we revealed that ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) is a target for S-mercuration in human neuroblastoma SH-SY5Y cells exposed to MeHg (1 µM, 9 hr). The modification site of UCH-L1 by MeHg was Cys152, as determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. MeHg was shown to inhibit the catalytic activity of recombinant human UCH-L1 in a concentration-dependent manner. Knockdown of UCH-L1 indicated that this enzyme plays a critical role in regulating mono-ubiquitin (monoUb) levels in SH-SY5Y cells and exposure of SH-SY5Y cells to MeHg caused a reduction in the level of monoUb in these cells. These observations suggest that UCH-L1 readily undergoes S-mercuration by MeHg through Cys152 and this covalent modification inhibits UCH-L1, leading to the potential disruption of the maintenance of cellular monoUb levels.

  7. Dihydrofolate Reductase Activity in Strains of Streptococcus faecium var. durans Resistant to Methasquin and Amethopterin1

    PubMed Central

    Rader, Jeanne I.; Hutchison, Dorris J.

    1972-01-01

    Resistance to the antifolates methasquin and amethopterin has been studied in new strains of Streptococcus faecium var. durans. Two methasquin-resistant strains (SF/MQ, SF/MQT) and an amethopterin-resistant strain (SF/AM) were selected independently from the wild-type S. faecium var. durans (SF/O). SF/MQT is a thymine auxotroph. Total dihydrofolate reductase activity was elevated in each of the resistant strains. The greatest increase (36-fold) was observed in extracts of SF/AM. The methasquin-resistant strains, SF/MQ and SF/MQT, had 29-fold and 8-fold, respectively, more dihydrofolate reductase activity than the parental strain. Total dihydrofolate reductase activity of SF/O was separable by gel filtration into two components: a folate reductase (11%) and a specific dihydrofolate reductase (89%). Folate reductase activity was associated with 88% of the total dihydrofolate reductase activity of SF/MQT, with specific dihydrofolate reductase activity accounting for the remaining 12%. In SF/MQ and SF/AM, folate reductase activity was associated with 97% of the total dihydrofolate reductase activity. Studies of the inhibition by methasquin and amethopterin of partially purified folate reductase and specific dihydrofolate reductase of the mutant strains suggested that resistance was not accompanied by changes in the affinities of these enzymes for either antifolate. PMID:4401600

  8. Adaptation of cytochrome-b5 reductase activity and methaemoglobinaemia in areas with a high nitrate concentration in drinking-water.

    PubMed Central

    Gupta, S. K.; Gupta, R. C.; Seth, A. K.; Gupta, A. B.; Bassin, J. K.; Gupta, A.

    1999-01-01

    An epidemiological investigation was undertaken in India to assess the prevalence of methaemoglobinaemia in areas with high nitrate concentration in drinking-water and the possible association with an adaptation of cytochrome-b5 reductase. Five areas were selected, with average nitrate ion concentrations in drinking-water of 26, 45, 95, 222 and 459 mg/l. These areas were visited and house schedules were prepared in accordance with a statistically designed protocol. A sample of 10% of the total population was selected in each of the areas, matched for age and weight, giving a total of 178 persons in five age groups. For each subject, a detailed history was documented, a medical examination was conducted and blood samples were taken to determine methaemoglobin level and cytochrome-b5 reductase activity. Collected data were subjected to statistical analysis to test for a possible relationship between nitrate concentration, cytochrome-b5 reductase activity and methaemoglobinaemia. High nitrate concentrations caused methaemoglobinaemia in infants and adults. The reserve of cytochrome-b5 reductase activity (i.e. the enzyme activity not currently being used, but which is available when needed; for example, under conditions of increased nitrate ingestion) and its adaptation with increasing water nitrate concentration to reduce methaemoglobin were more pronounced in children and adolescents. PMID:10534899

  9. Adaptation of cytochrome-b5 reductase activity and methaemoglobinaemia in areas with a high nitrate concentration in drinking-water.

    PubMed

    Gupta, S K; Gupta, R C; Seth, A K; Gupta, A B; Bassin, J K; Gupta, A

    1999-01-01

    An epidemiological investigation was undertaken in India to assess the prevalence of methaemoglobinaemia in areas with high nitrate concentration in drinking-water and the possible association with an adaptation of cytochrome-b5 reductase. Five areas were selected, with average nitrate ion concentrations in drinking-water of 26, 45, 95, 222 and 459 mg/l. These areas were visited and house schedules were prepared in accordance with a statistically designed protocol. A sample of 10% of the total population was selected in each of the areas, matched for age and weight, giving a total of 178 persons in five age groups. For each subject, a detailed history was documented, a medical examination was conducted and blood samples were taken to determine methaemoglobin level and cytochrome-b5 reductase activity. Collected data were subjected to statistical analysis to test for a possible relationship between nitrate concentration, cytochrome-b5 reductase activity and methaemoglobinaemia. High nitrate concentrations caused methaemoglobinaemia in infants and adults. The reserve of cytochrome-b5 reductase activity (i.e. the enzyme activity not currently being used, but which is available when needed; for example, under conditions of increased nitrate ingestion) and its adaptation with increasing water nitrate concentration to reduce methaemoglobin were more pronounced in children and adolescents.

  10. Ion source

    DOEpatents

    Leung, Ka-Ngo; Ehlers, Kenneth W.

    1984-01-01

    A magnetic filter for an ion source reduces the production of undesired ion species and improves the ion beam quality. High-energy ionizing electrons are confined by the magnetic filter to an ion source region, where the high-energy electrons ionize gas molecules. One embodiment of the magnetic filter uses permanent magnets oriented to establish a magnetic field transverse to the direction of travel of ions from the ion source region to the ion extraction region. In another embodiment, low energy 16 eV electrons are injected into the ion source to dissociate gas molecules and undesired ion species into desired ion species.

  11. ARSENICALS INHIBIT THIOREDOXIN REDUCTASE ACTIVITY IN CULTURED RAT HEPATOCYTES

    EPA Science Inventory

    ARSENICALS INHIBIT THIOREDOXIN REDUCTASE ACTIVITY IN CULTURED RAT HEPATOCYTES.

    S. Lin1, L. M. Del Razo1, M. Styblo1, C. Wang2, W. R. Cullen2, and D.J. Thomas3. 1Univ. North Carolina, Chapel Hill, NC; 2Univ. British Columbia, Vancouver, BC, Canada; 3National Health and En...

  12. 21 CFR 864.7375 - Glutathione reductase assay.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Glutathione reductase assay. 864.7375 Section 864.7375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7375...

  13. 21 CFR 864.7375 - Glutathione reductase assay.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Glutathione reductase assay. 864.7375 Section 864.7375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7375 Glutathione...

  14. 21 CFR 864.7375 - Glutathione reductase assay.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Glutathione reductase assay. 864.7375 Section 864.7375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7375 Glutathione...

  15. 21 CFR 864.7375 - Glutathione reductase assay.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Glutathione reductase assay. 864.7375 Section 864.7375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7375 Glutathione...

  16. 21 CFR 864.7375 - Glutathione reductase assay.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Glutathione reductase assay. 864.7375 Section 864.7375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7375 Glutathione...

  17. Obtaining partial purified xylose reductase from Candida guilliermondii

    PubMed Central

    Tomotani, Ester Junko; de Arruda, Priscila Vaz; Vitolo, Michele; de Almeida Felipe, Maria das Graças

    2009-01-01

    The enzymatic bioconversion of xylose into xylitol by xylose reductase (XR) is an alternative for chemical and microbiological processes. The partial purified XR was obtained by using the following three procedures: an agarose column, a membrane reactor or an Amicon Ultra-15 50K Centrifugal Filter device at yields of 40%, 7% and 67%, respectively. PMID:24031408

  18. A detoxifying oxygen reductase in the anaerobic protozoan Entamoeba histolytica.

    PubMed

    Vicente, João B; Tran, Vy; Pinto, Liliana; Teixeira, Miguel; Singh, Upinder

    2012-09-01

    We report the characterization of a bacterial-type oxygen reductase abundant in the cytoplasm of the anaerobic protozoan parasite Entamoeba histolytica. Upon host infection, E. histolytica is confronted with various oxygen tensions in the host intestine, as well as increased reactive oxygen and nitrogen species at the site of local tissue inflammation. Resistance to oxygen-derived stress thus plays an important role in the pathogenic potential of E. histolytica. The genome of E. histolytica has four genes that encode flavodiiron proteins, which are bacterial-type oxygen or nitric oxide reductases and were likely acquired by lateral gene transfer from prokaryotes. The EhFdp1 gene has higher expression in virulent than in nonvirulent Entamoeba strains and species, hinting that the response to oxidative stress may be one correlate of virulence potential. We demonstrate that EhFdp1 is abundantly expressed in the cytoplasm of E. histolytica and that the protein levels are markedly increased (up to ~5-fold) upon oxygen exposure. Additionally, we produced fully functional recombinant EhFdp1 and demonstrated that this enzyme is a specific and robust oxygen reductase but has poor nitric oxide reductase activity. This observation represents a new mechanism of oxygen resistance in the anaerobic protozoan pathogen E. histolytica.

  19. The arsenic hyperaccumulating Pteris vittata expresses two arsenate reductases.

    PubMed

    Cesaro, Patrizia; Cattaneo, Chiara; Bona, Elisa; Berta, Graziella; Cavaletto, Maria

    2015-09-28

    Enzymatic reduction of arsenate to arsenite is the first known step in arsenate metabolism in all organisms. Although the presence of one mRNA arsenate reductase (PvACR2) has been characterized in gametophytes of P. vittata, no arsenate reductase protein has been directly observed in this arsenic hyperaccumulating fern, yet. In order to assess the possible presence of arsenate reductase in P. vittata, two recombinant proteins, ACR2-His6 and Trx-His6-S-Pv2.5-8 were prepared in Escherichia coli, purified and used to produce polyclonal antibodies. The presence of these two enzymes was evaluated by qRT-PCR, immunoblotting and direct MS analysis. Enzymatic activity was detected in crude extracts. For the first time we detected and identified two arsenate reductase proteins (PvACR2 and Pv2.5-8) in sporophytes and gametophytes of P. vittata. Despite an increase of the mRNA levels for both proteins in roots, no difference was observed at the protein level after arsenic treatment. Overall, our data demonstrate the constitutive protein expression of PvACR2 and Pv2.5-8 in P. vittata tissues and propose their specific role in the complex metabolic network of arsenic reduction.

  20. Dissimilatory Nitrite Reductase Genes from Autotrophic Ammonia-Oxidizing Bacteria

    PubMed Central

    Casciotti, Karen L.; Ward, Bess B.

    2001-01-01

    The presence of a copper-containing dissimilatory nitrite reductase gene (nirK) was discovered in several isolates of β-subdivision ammonia-oxidizing bacteria using PCR and DNA sequencing. PCR primers Cunir3 and Cunir4 were designed based on published nirK sequences from denitrifying bacteria and used to amplify a 540-bp fragment of the nirK gene from Nitrosomonas marina and five additional isolates of ammonia-oxidizing bacteria. Amplification products of the expected size were cloned and sequenced. Alignment of the nucleic acid and deduced amino acid (AA) sequences shows significant similarity (62 to 75% DNA, 58 to 76% AA) between nitrite reductases present in these nitrifiers and the copper-containing nitrite reductase found in classic heterotrophic denitrifiers. While the presence of a nitrite reductase in Nitrosomonas europaea is known from early biochemical work, preliminary sequence data from its genome indicate a rather low similarity to the denitrifier nirKs. Phylogenetic analysis of the partial nitrifier nirK sequences indicates that the topology of the nirK tree corresponds to the 16S rRNA and amoA trees. While the role of nitrite reduction in the metabolism of nitrifying bacteria is still uncertain, these data show that the nirK gene is present in closely related nitrifying isolates from many oceanographic regions and suggest that nirK sequences retrieved from the environment may include sequences from ammonia-oxidizing bacteria. PMID:11319103

  1. The arsenic hyperaccumulating Pteris vittata expresses two arsenate reductases

    PubMed Central

    Cesaro, Patrizia; Cattaneo, Chiara; Bona, Elisa; Berta, Graziella; Cavaletto, Maria

    2015-01-01

    Enzymatic reduction of arsenate to arsenite is the first known step in arsenate metabolism in all organisms. Although the presence of one mRNA arsenate reductase (PvACR2) has been characterized in gametophytes of P. vittata, no arsenate reductase protein has been directly observed in this arsenic hyperaccumulating fern, yet. In order to assess the possible presence of arsenate reductase in P. vittata, two recombinant proteins, ACR2-His6 and Trx-His6-S-Pv2.5–8 were prepared in Escherichia coli, purified and used to produce polyclonal antibodies. The presence of these two enzymes was evaluated by qRT-PCR, immunoblotting and direct MS analysis. Enzymatic activity was detected in crude extracts. For the first time we detected and identified two arsenate reductase proteins (PvACR2 and Pv2.5–8) in sporophytes and gametophytes of P. vittata. Despite an increase of the mRNA levels for both proteins in roots, no difference was observed at the protein level after arsenic treatment. Overall, our data demonstrate the constitutive protein expression of PvACR2 and Pv2.5–8 in P. vittata tissues and propose their specific role in the complex metabolic network of arsenic reduction. PMID:26412036

  2. ARSENICALS INHIBIT THIOREDOXIN REDUCTASE ACTIVITY IN CULTURED RAT HEPATOCYTES

    EPA Science Inventory

    ARSENICALS INHIBIT THIOREDOXIN REDUCTASE ACTIVITY IN CULTURED RAT HEPATOCYTES.

    S. Lin1, L. M. Del Razo1, M. Styblo1, C. Wang2, W. R. Cullen2, and D.J. Thomas3. 1Univ. North Carolina, Chapel Hill, NC; 2Univ. British Columbia, Vancouver, BC, Canada; 3National Health and En...

  3. The Kinetics and Inhibition of the Enzyme Methemoglobin Reductase

    ERIC Educational Resources Information Center

    Splittgerber, A. G.; And Others

    1975-01-01

    Describes an undergraduate biochemistry experiment which involves the preparation and kinetics of an oxidation-reduction enzyme system, methemoglobin reductase. A crude enzyme extract is prepared and assayed spectrophotometrically. The enzyme system obeys Michaelis-Menton kinetics with respect to both substrate and the NADH cofactor. (MLH)

  4. Dihydrofolate reductase: A potential drug target in trypanosomes and leishmania

    NASA Astrophysics Data System (ADS)

    Zuccotto, Fabio; Martin, Andrew C. R.; Laskowski, Roman A.; Thornton, Janet M.; Gilbert, Ian H.

    1998-05-01

    Dihydrofolate reductase has successfully been used as a drug target in the area of anti-cancer, anti-bacterial and anti-malarial chemotherapy. Little has been done to evaluate it as a drug target for treatment of the trypanosomiases and leishmaniasis. A crystal structure of Leishmania major dihydrofolate reductase has been published. In this paper, we describe the modelling of Trypanosoma cruzi and Trypanosoma brucei dihydrofolate reductases based on this crystal structure. These structures and models have been used in the comparison of protozoan, bacterial and human enzymes in order to highlight the different features that can be used in the design of selective anti-protozoan agents. Comparison has been made between residues present in the active site, the accessibility of these residues, charge distribution in the active site, and the shape and size of the active sites. Whilst there is a high degree of similarity between protozoan, human and bacterial dihydrofolate reductase active sites, there are differences that provide potential for selective drug design. In particular, we have identified a set of residues which may be important for selective drug design and identified a larger binding pocket in the protozoan than the human and bacterial enzymes.

  5. Studies on Marek's Disease Virus Encoded Ribonucleotide Reductase

    USDA-ARS?s Scientific Manuscript database

    Ribonucleotide reductase (RR) is an essential enzyme for the conversion of ribonucleotides to deoxyribonucleotides in prokaryotic and eukaryotic cells. The enzyme consists of two subunits namely RR1 and RR2, both of which associate to form an active holoenzyme. Herpesviruses express a functional R...

  6. [Malate oxidation by mitochondrial succinate:ubiquinone-reductase].

    PubMed

    Belikova, Iu O; Kotliar, A B

    1988-04-01

    Succinate:ubiquinone reductase was shown to catalyze the oxidation of L- and D-stereoisomers of malate by artificial electron acceptors and ubiquinone. The rate of malate oxidation by succinate:ubiquinone reductase is by two orders of magnitude lower than that for the natural substrate--succinate. The values of kinetic constants for the oxidation of D- and L-stereoisomers of malate are equal to: V infinity = 0.1 mumol/min/mg protein, Km = 2 mM and V infinity = 0.05 mumol/min/mg protein, Km = 2 mM, respectively. The malate dehydrogenase activity is fully inhibited by the inhibitors of the dicarboxylate-binding site of the enzyme, i.e., N-ethylmaleimide and malonate and is practically insensitive to carboxin, a specific inhibitor of the ubiquinone-binding center. The enol form of oxaloacetate was shown to be the product of malate oxidation by succinate:ubiquinone reductase. The kinetics of inhibition of the enzyme activity by the ketone and enol forms of oxaloacetate was studied. Both forms of oxaloacetate effectively inhibit the succinate:ubiquinone reductase reaction.

  7. Molecular genetics of steroid 5 alpha-reductase 2 deficiency.

    PubMed Central

    Thigpen, A E; Davis, D L; Milatovich, A; Mendonca, B B; Imperato-McGinley, J; Griffin, J E; Francke, U; Wilson, J D; Russell, D W

    1992-01-01

    Two isozymes of steroid 5 alpha-reductase encoded by separate loci catalyze the conversion of testosterone to dihydrotestosterone. Inherited defects in the type 2 isozyme lead to male pseudohermaphroditism in which affected males have a normal internal urogenital tract but external genitalia resembling those of a female. The 5 alpha-reductase type 2 gene (gene symbol SRD5A2) was cloned and shown to contain five exons and four introns. The gene was localized to chromosome 2 band p23 by somatic cell hybrid mapping and chromosomal in situ hybridization. Molecular analysis of the SRD5A2 gene resulted in the identification of 18 mutations in 11 homozygotes, 6 compound heterozygotes, and 4 inferred compound heterozygotes from 23 families with 5 alpha-reductase deficiency. 6 apparent recurrent mutations were detected in 19 different ethnic backgrounds. In two patients, the catalytic efficiency of the mutant enzymes correlated with the severity of the disease. The high proportion of compound heterozygotes suggests that the carrier frequency of mutations in the 5 alpha-reductase type 2 gene may be higher than previously thought. Images PMID:1522235

  8. Molybdenum-containing nitrite reductases: Spectroscopic characterization and redox mechanism.

    PubMed

    Wang, Jun; Keceli, Gizem; Cao, Rui; Su, Jiangtao; Mi, Zhiyuan

    2017-01-01

    This review summarizes the spectroscopic results, which will provide useful suggestions for future research. In addition, the fields that urgently need more information are also advised. Nitrite-NO-cGMP has been considered as an important signaling pathway of NO in human cells. To date, all the four known human molybdenum-containing enzymes, xanthine oxidase, aldehyde oxidase, sulfite oxidase, and mitochondrial amidoxime-reducing component, have been shown to function as nitrite reductases under hypoxia by biochemical, cellular, or animal studies. Various spectroscopic techniques have been applied to investigate the structure and catalytic mechanism of these enzymes for more than 20 years. We summarize the published data on the applications of UV-vis and EPR spectroscopies, and X-ray crystallography in studying nitrite reductase activity of the four human molybdenum-containing enzymes. UV-vis has provided useful information on the redox active centers of these enzymes. The utilization of EPR spectroscopy has been critical in determining the coordination and redox status of the Mo center during catalysis. Despite the lack of substrate-bound crystal structures of these nitrite reductases, valuable structural information has been obtained by X-ray crystallography. To fully understand the catalytic mechanisms of these physiologically/pathologically important nitrite reductases, structural studies on substrate-redox center interaction are needed.

  9. The Kinetics and Inhibition of the Enzyme Methemoglobin Reductase

    ERIC Educational Resources Information Center

    Splittgerber, A. G.; And Others

    1975-01-01

    Describes an undergraduate biochemistry experiment which involves the preparation and kinetics of an oxidation-reduction enzyme system, methemoglobin reductase. A crude enzyme extract is prepared and assayed spectrophotometrically. The enzyme system obeys Michaelis-Menton kinetics with respect to both substrate and the NADH cofactor. (MLH)

  10. Thioredoxin and NADP-thioredoxin reductase from cultured carrot cells

    NASA Technical Reports Server (NTRS)

    Johnson, T. C.; Cao, R. Q.; Kung, J. E.; Buchanan, B. B.

    1987-01-01

    Dark-grown carrot (Daucus carota L.) tissue cultures were found to contain both protein components of the NADP/thioredoxin system--NADP-thioredoxin reductase and the thioredoxin characteristic of heterotrophic systems, thioredoxin h. Thioredoxin h was purified to apparent homogeneity and, like typical bacterial counterparts, was a 12-kdalton (kDa) acidic protein capable of activating chloroplast NADP-malate dehydrogenase (EC 1.1.1.82) more effectively than fructose-1,6-bisphosphatase (EC 3.1.3.11). NADP-thioredoxin reductase (EC 1.6.4.5) was partially purified and found to be an arsenite-sensitive enzyme composed of two 34-kDa subunits. Carrot NADP-thioredoxin reductase resembled more closely its counterpart from bacteria rather than animal cells in acceptor (thioredoxin) specificity. Upon greening of the cells, the content of NADP-thioredoxin-reductase activity, and, to a lesser extent, thioredoxin h decreased. The results confirm the presence of a heterotrophic-type thioredoxin system in plant cells and raise the question of its physiological function.

  11. 5. cap alpha. -reductase activity in rat adipose tissue

    SciTech Connect

    Zyirek, M.; Flood, C.; Longcope, C.

    1987-11-01

    We measured the 5 ..cap alpha..-reductase activity in isolated cell preparations of rat adipose tissue using the formation of (/sup 3/H) dihydrotestosterone from (/sup 3/H) testosterone as an endpoint. Stromal cells were prepared from the epididymal fat pad, perinephric fat, and subcutaneous fat of male rats and from perinephric fat of female rats. Adipocytes were prepared from the epididymal fat pad and perinephric fat of male rats. Stromal cells from the epididymal fat pad and perinephric fat contained greater 5..cap alpha..-reductase activity than did the adipocytes from these depots. Stromal cells from the epididymal fat pad contained greater activity than those from perinephric and subcutaneous depots. Perinephric stromal cells from female rats were slightly more active than those from male rats. Estradiol (10/sup -8/ M), when added to the medium, caused a 90% decrease in 5..cap alpha..-reductase activity. Aromatase activity was minimal, several orders of magnitude less than 5..cap alpha..-reductase activity in each tissue studied.

  12. Characterization of mitochondrial thioredoxin reductase from C. elegans

    SciTech Connect

    Lacey, Brian M.; Hondal, Robert J. . E-mail: Robert.Hondal@uvm.edu

    2006-08-04

    Thioredoxin reductase catalyzes the NADPH-dependent reduction of the catalytic disulfide bond of thioredoxin. In mammals and other higher eukaryotes, thioredoxin reductases contain the rare amino acid selenocysteine at the active site. The mitochondrial enzyme from Caenorhabditis elegans, however, contains a cysteine residue in place of selenocysteine. The mitochondrial C. elegans thioredoxin reductase was cloned from an expressed sequence tag and then produced in Escherichia coli as an intein-fusion protein. The purified recombinant enzyme has a k {sub cat} of 610 min{sup -1} and a K {sub m} of 610 {mu}M using E. coli thioredoxin as substrate. The reported k {sub cat} is 25% of the k {sub cat} of the mammalian enzyme and is 43-fold higher than a cysteine mutant of mammalian thioredoxin reductase. The enzyme would reduce selenocysteine, but not hydrogen peroxide or insulin. The flanking glycine residues of the GCCG motif were mutated to serine. The mutants improved substrate binding, but decreased the catalytic rate.

  13. The arsenic hyperaccumulating Pteris vittata expresses two arsenate reductases

    NASA Astrophysics Data System (ADS)

    Cesaro, Patrizia; Cattaneo, Chiara; Bona, Elisa; Berta, Graziella; Cavaletto, Maria

    2015-09-01

    Enzymatic reduction of arsenate to arsenite is the first known step in arsenate metabolism in all organisms. Although the presence of one mRNA arsenate reductase (PvACR2) has been characterized in gametophytes of P. vittata, no arsenate reductase protein has been directly observed in this arsenic hyperaccumulating fern, yet. In order to assess the possible presence of arsenate reductase in P. vittata, two recombinant proteins, ACR2-His6 and Trx-His6-S-Pv2.5-8 were prepared in Escherichia coli, purified and used to produce polyclonal antibodies. The presence of these two enzymes was evaluated by qRT-PCR, immunoblotting and direct MS analysis. Enzymatic activity was detected in crude extracts. For the first time we detected and identified two arsenate reductase proteins (PvACR2 and Pv2.5-8) in sporophytes and gametophytes of P. vittata. Despite an increase of the mRNA levels for both proteins in roots, no difference was observed at the protein level after arsenic treatment. Overall, our data demonstrate the constitutive protein expression of PvACR2 and Pv2.5-8 in P. vittata tissues and propose their specific role in the complex metabolic network of arsenic reduction.

  14. [Inhibition of aldose reductase by Chinese herbal medicine].

    PubMed

    Mao, X M; Zhang, J Q

    1993-10-01

    Seven Chinese herbal drugs were screened for experimental inhibition of lens aldose reductase activity, among which quercetin exhibited potent enzyme-inhibitory activities in vitro. Its IC50 value was 3.44 x 10(-7) mol/L. It may be helpful in the prophylaxis and treatment of diabetic complications.

  15. Thioredoxin and NADP-thioredoxin reductase from cultured carrot cells

    NASA Technical Reports Server (NTRS)

    Johnson, T. C.; Cao, R. Q.; Kung, J. E.; Buchanan, B. B.

    1987-01-01

    Dark-grown carrot (Daucus carota L.) tissue cultures were found to contain both protein components of the NADP/thioredoxin system--NADP-thioredoxin reductase and the thioredoxin characteristic of heterotrophic systems, thioredoxin h. Thioredoxin h was purified to apparent homogeneity and, like typical bacterial counterparts, was a 12-kdalton (kDa) acidic protein capable of activating chloroplast NADP-malate dehydrogenase (EC 1.1.1.82) more effectively than fructose-1,6-bisphosphatase (EC 3.1.3.11). NADP-thioredoxin reductase (EC 1.6.4.5) was partially purified and found to be an arsenite-sensitive enzyme composed of two 34-kDa subunits. Carrot NADP-thioredoxin reductase resembled more closely its counterpart from bacteria rather than animal cells in acceptor (thioredoxin) specificity. Upon greening of the cells, the content of NADP-thioredoxin-reductase activity, and, to a lesser extent, thioredoxin h decreased. The results confirm the presence of a heterotrophic-type thioredoxin system in plant cells and raise the question of its physiological function.

  16. The polymorphisms in methylenetetrahydrofolate reductase, methionine synthase, methionine synthase reductase, and the risk of colorectal cancer.

    PubMed

    Zhou, Daijun; Mei, Qiang; Luo, Han; Tang, Bo; Yu, Peiwu

    2012-01-01

    Polymorphisms in genes involved in folate metabolism may modulate the risk of colorectal cancer (CRC), but data from published studies are conflicting. The current meta-analysis was performed to address a more accurate estimation. A total of 41 (17,552 cases and 26,238 controls), 24(8,263 cases and 12,033 controls), 12(3,758 cases and 5,646 controls), and 13 (5,511 cases and 7,265 controls) studies were finally included for the association between methylenetetrahydrofolate reductase (MTHFR) C677T and A1289C, methione synthase reductase (MTRR) A66G, methionine synthase (MTR) A2756G polymorphisms and the risk of CRC, respectively. The data showed that the MTHFR 677T allele was significantly associated with reduced risk of CRC (OR = 0.93, 95%CI 0.90-0.96), while the MTRR 66G allele was significantly associated with increased risk of CRC (OR = 1.11, 95%CI 1.01-1.18). Sub-group analysis by ethnicity revealed that MTHFR C677T polymorphism was significantly associated with reduced risk of CRC in Asians (OR = 0.80, 95%CI 0.72-0.89) and Caucasians (OR = 0.84, 95%CI 0.76-0.93) in recessive genetic model, while the MTRR 66GG genotype was found to significantly increase the risk of CRC in Caucasians (GG vs. AA: OR = 1.18, 95%CI 1.03-1.36). No significant association was found between MTHFR A1298C and MTR A2756G polymorphisms and the risk of CRC. Cumulative meta-analysis showed no particular time trend existed in the summary estimate. Probability of publication bias was low across all comparisons illustrated by the funnel plots and Egger's test. Collectively, this meta-analysis suggested that MTHFR 677T allele might provide protection against CRC in worldwide populations, while MTRR 66G allele might increase the risk of CRC in Caucasians. Since potential confounders could not be ruled out completely, further studies were needed to confirm these results.

  17. The Polymorphisms in Methylenetetrahydrofolate Reductase, Methionine Synthase, Methionine Synthase Reductase, and the Risk of Colorectal Cancer

    PubMed Central

    Zhou, Daijun; Mei, Qiang; Luo, Han; Tang, Bo; Yu, Peiwu

    2012-01-01

    Polymorphisms in genes involved in folate metabolism may modulate the risk of colorectal cancer (CRC), but data from published studies are conflicting. The current meta-analysis was performed to address a more accurate estimation. A total of 41 (17,552 cases and 26,238 controls), 24(8,263 cases and 12,033 controls), 12(3,758 cases and 5,646 controls), and 13 (5,511 cases and 7,265 controls) studies were finally included for the association between methylenetetrahydrofolate reductase (MTHFR) C677T and A1289C, methione synthase reductase (MTRR) A66G, methionine synthase (MTR) A2756G polymorphisms and the risk of CRC, respectively. The data showed that the MTHFR 677T allele was significantly associated with reduced risk of CRC (OR = 0.93, 95%CI 0.90-0.96), while the MTRR 66G allele was significantly associated with increased risk of CRC (OR = 1.11, 95%CI 1.01-1.18). Sub-group analysis by ethnicity revealed that MTHFR C677T polymorphism was significantly associated with reduced risk of CRC in Asians (OR = 0.80, 95%CI 0.72-0.89) and Caucasians (OR = 0.84, 95%CI 0.76-0.93) in recessive genetic model, while the MTRR 66GG genotype was found to significantly increase the risk of CRC in Caucasians (GG vs. AA: OR = 1.18, 95%CI 1.03-1.36). No significant association was found between MTHFR A1298C and MTR A2756G polymorphisms and the risk of CRC. Cumulative meta-analysis showed no particular time trend existed in the summary estimate. Probability of publication bias was low across all comparisons illustrated by the funnel plots and Egger's test. Collectively, this meta-analysis suggested that MTHFR 677T allele might provide protection against CRC in worldwide populations, while MTRR 66G allele might increase the risk of CRC in Caucasians. Since potential confounders could not be ruled out completely, further studies were needed to confirm these results. PMID:22719222

  18. Ion-Ion Neutralization.

    DTIC Science & Technology

    1980-12-31

    ion flow tube (SIFT) experiments have been given in previous publications and so only the essential features and tho.;e detail:. specific to the present...rapidly, essentially at the gas kinetic limiting rate within the accuracy of the experimental data (1 30% on the measured rate coefficients). The...the various negative ions remained essentially invariant along the length of the plasma column. The data in Table C show that ". for all of the

  19. Comparative Studies on the Induction and Inactivation of Nitrate Reductase in Corn Roots and Leaves 1

    PubMed Central

    Aslam, Muhammad; Oaks, Ann

    1976-01-01

    A comparison of induction and inactivation of nitrate reductase and two of its component activities, namely FMNH2-nitrate reductase and NO3−-induced NADH-cytochrome c reductase, was made in roots and leaves of corn (Zea mays L. var. W64A × 182E). The three activities were induced in parallel in both tissues when NO3− was supplied. WO4= suppressed the induction of NADH- and FMNH2-nitrate reductase activities in root tips and leaves. The NO3−-induced NADH-cytochrome c reductase activity showed a normal increase in roots treated with WO4=. In leaves, on the other hand, there was a marked superinduction of the NO3−-induced NADH-cytochrome c reductase in the presence of WO4=. The half-life values of NADH-nitrate reductase and FMNH2-nitrate reductase measured by removing NO3− and adding WO4= to the medium, were 4 hours in root tips and 6 hours in excised leaves. Addition of NO3− in the induction medium together with WO4= gave partial protection of NADH-nitrate reductase and FMNH2-nitrate reductase activities in both root tips and leaves with a t0.5 of 6 and 8 hours, respectively. NO3− also reduced the loss of nitrate reductase activity from mature root sections. In the presence of cycloheximide, both NADH-nitrate reductase and NO3−-induced NADH-cytochrome c reductase activities were lost at similar rates in root tips. NO3− protected the loss of NO3−-induced NADH-cytochrome c reductase to the same extent as that of NADH-nitrate reductase. PMID:16659529

  20. Measurement of nitrous oxide reductase activity in aquatic sediments

    SciTech Connect

    Miller, L.G.; Oremland, R.S.; Paulsen, S.

    1986-01-01

    Denitrification in aquatic sediments was measured by an N/sub 2/O reductase assay. Sediments consumed small added quantities of N/sub 2/O over short periods (a few hours). In experiments with sediment slurries, N/sub 2/O reductase activity was inhibited by 0/sub 2/, C/sub 2/H/sub 2/, heat treatment, and by high levels of nitrate (1 mM) or sulfide (10 mM). However, ambient levels of nitrate (<100 ..mu..M) did not influence activity, and moderate levels (about 150 ..mu..M) induced only a short lag before reductase activity began. Moderate levels of sulfide (<1 mM) had no effect on N/sub 2/O reductase activity. Nitrous oxide reductase displayed Michaelis-Menten kinetics in sediments from freshwater, estuarine, and alkaline-saline environments. An in situ assay was devised in which a solution of N/sub 2/O was injected into sealed glass cores containing intact sediment. Two estimates of net rates of denitrification in San Francisco Bay under approximated in situ conditions were 0.009 and 0.041 mmol of N/sub 2/O per m/sup 2/ per h. Addition of chlorate to inhibit denitrification in these intact-core experiments (to estimate gross rates of N/sub 2/O consumption) resulted in approximately a 14% upward revision of estimates of net rates. These results were comparable to an in situ estimate of 0.022 mmol of N/sub 2/O per m/sup 2/ per h made with the acetylene block assay.

  1. Measurement of nitrous oxide reductase activity in aquatic sediments

    USGS Publications Warehouse

    Miller, L.G.; Oremland, R.S.; Paulsen, S.

    1986-01-01

    Denitrification in aquatic sediments was measured by an N2O reductase assay. Sediments consumed small added quantities of N2O over short periods (a few hours). In experiments with sediment slurries, N2O reductase activity was inhibited by O2, C2H2, heat treatment, and by high levels of nitrate (1 mM) or sulfide (10 mM). However, ambient levels of nitrate (<100 μM) did not influence activity, and moderate levels (about 150 μM) induced only a short lag before reductase activity began. Moderate levels of sulfide (<1 mM) had no effect on N2O reductase activity. Nitrous oxide reductase displayed Michaelis-Menten kinetics in sediments from freshwater (Km = 2.17 μM), estuarine (Km = 14.5 μM), and alkaline-saline (Km = 501 μM) environments. An in situ assay was devised in which a solution of N2O was injected into sealed glass cores containing intact sediment. Two estimates of net rates of denitrification in San Francisco Bay under approximated in situ conditions were 0.009 and 0.041 mmol of N2O per m2 per h. Addition of chlorate to inhibit denitrification in these intact-core experiments (to estimate gross rates of N2O consumption) resulted in approximately a 14% upward revision of estimates of net rates. These results were comparable to an in situ estimate of 0.022 mmol of N2O per m2 per h made with the acetylene block assay.

  2. Identification of 5α-reductase isoenzymes in canine skin.

    PubMed

    Bernardi de Souza, Lucilene; Paradis, Manon; Zamberlam, Gustavo; Benoit-Biancamano, Marie-Odile; Price, Christopher

    2015-10-01

    Alopecia X in dogs is a noninflammatory alopecia that may be caused by a hormonal dysfunction. It may be similar to androgenic alopecia in men that is caused by the effect of dihydrotestosterone (DHT). The 5α-reductase isoenzymes, 5αR1 and 5αR2, and a recently described 5αR3, are responsible for the conversion of testosterone into DHT. However, which 5α-reductases are present in canine skin has not yet been described. The main objective of this study was to determine the pattern of expression of 5α-reductase genes in canine skin. Skin biopsies were obtained from healthy, intact young-mature beagles (three males, four females) at three anatomical sites normally affected by alopecia X (dorsal neck, back of thighs and base of tail) and two sites generally unaffected (dorsal head and ventral thorax). Prostate samples (n = 3) were collected as positive controls for 5α-reductase mRNA abundance measurement by real-time PCR. We detected mRNA encoding 5αR1 and 5αR3 but not 5αR2. There were no significant differences in 5αR1 and 5αR3 mRNA levels between the different anatomical sites, irrespective of gender (P > 0.05). Moreover, the mean mRNA abundance in each anatomical site did not differ between males and females (P > 0.05). To the best of the authors' knowledge, this is the first study demonstrating the expression of 5α-reductases in canine skin and the expression of 5αR3 in this tissue. These results may help to elucidate the pathogenesis of alopecia X and to determine more appropriate treatments for this disorder. © 2015 ESVD and ACVD.

  3. Leukemia L1210 cell lines resistant to ribonucleotide reductase inhibitors.

    PubMed

    Cory, J G; Carter, G L

    1988-02-15

    Leukemia L1210 cell lines, ED1 and ED2, were generated which were resistant to the cytotoxic effects of deoxyadenosine/erythro-9-(2-hydroxyl-3-nonyl)adenine and deoxyadenosine/erythro-9-(2-hydroxyl-3-nonyl)adenine plus 2,3-dihydro-1H-pyrazole[2,3a]imidazole/Desferal, respectively. The ED1 and ED2 were characterized to show that these cell lines had increased levels of ribonucleotide reductase as measured by CDP reduction. The reductase activity in crude cell-free extracts from the ED1 and ED2 cells was not inhibited by dATP. For CDP reductase, the activation by adenylylimido diphosphate and inhibition by dGTP and dTTP in these extracts from the ED1 and ED2 cells were the same as for the wild-type L1210 cells. The ED1 and ED2 cells were highly cross-resistant, as measured by growth inhibition, to deoxyguanosine/8-aminoguanosine, 2-fluorodeoxyadenosine, and 2-fluoroadenine arabinoside. While the ED2 cells showed resistance to 2,3-dihydro-1H-pyrazole-[2,3a]-imidazole/Desferal (6-fold), the ED1 and ED2 cell lines showed less resistance to hydroxyurea, 4-methyl-5-amino-1-formylisoquinoline thiosemicarbazone, and the dialdehyde of inosine. These data indicate that the mechanisms of resistance to the ribonucleotide reductase inhibitors are related to the increased level of ribonucleotide reductase activity and to the decreased sensitivity of the effector-binding subunit to dATP.

  4. Assimilatory nitrate reductase from the green alga Ankistrodesmus braunii.

    PubMed

    De la Rosa, M A

    1983-01-01

    Assimilatory nitrate reductase (NAD(P)H-nitrate oxidoreductase, EC 1.6.6.2) from the green alga Ankistrodesmus braunii can be purified to homogeneity by dye-ligand chromatography on blue-Sepharose. The purified enzyme, whose turnover number is 623 s-1, presents an optimum pH of 7.5 and Km values of 13 microM, 23 microM and 0.15 mM for NADH, NADPH and nitrate, respectively. The NADH-nitrate reductase activity exhibits an iso ping pong bi bi kinetic mechanism. The molecular weight of the native nitrate reductase is 467 400, while that of its subunits is 58 750. These values suggest an octameric structure for the enzyme, which has been confirmed by electron microscopy. As deduced from spectrophotometric and fluorimetric studies, the enzyme contains FAD and cytochrome b-557 as prosthetic groups. FAD is not covalently bound to the protein and is easily dissociated in diluted solutions from the enzyme. Its apparent Km value is 4 nM, indicative of a high affinity of the enzyme for FAD. The results of the quantitative analyses of prosthetic groups indicate that nitrate reductase contains four molecules of flavin, four heme irons, and two atoms of molybdenum. The three components act sequentially transferring electrons from reduced pyridine nucleotides to nitrate, thus forming a short electron transport chain along the protein. A mechanism is proposed for the redox interconversion of the nitrate reductase activity. Inactivation seems to occur by formation of a stable complex of reduced enzyme with cyanide or superoxide, while reactivation is a consequence of reoxidation of the inactive enzyme. Both reactions imply the transfer of only one electron.

  5. Treatment of hirsutism with 5 alpha-reductase inhibitors.

    PubMed

    Brooks, J R

    1986-05-01

    Much os the evidence gathered from studies of 5 alpha-reductase activity levels and androgen metabolism in the skin of hirsute women and the excretion of androgen metabolites by hirsute women indicates that 5 alpha-reduced androgens are probably of primary importance in hirsutism. Unfortunately, until very recently, the lack of a suitable 5 alpha-reductase inhibitor made it very difficult to adequately test the hypothesis that such an inhibitor might be useful in the treatment of hirsutism and certain other androgen-related diseases. No substance was available which had good, unambiguous activity in vivo as a 5 alpha-reductase inhibitor. A number of 4-azasteroids have now been found to possess excellent 5 alpha-reductase inhibitory activity both in vitro and in vivo. Among other properties, several of these compounds show little or no affinity for the androgen receptor of rat prostate cytosol, they attenuate the growth promoting effect of T, but not DHT, on the ventral prostate of castrated male rats, they cause a marked reduction in prostatic DHT concentration in acutely treated rats and dogs and they bring about a significant decline in prostate size in chronically treated rats and dogs. It is expected that, in the near future, one or more of these highly active 5 alpha-reductase inhibitors will be tested in the clinic as a treatment for hirsutism. The results of those studies will be awaited with a great deal of interest since they should considerably advance our understanding of this disease and possibly contribute to its control.

  6. Crystal structures of pinoresinol-lariciresinol and phenylcoumaran benzylic ether reductases and their relationship to isoflavone reductases

    NASA Technical Reports Server (NTRS)

    Min, Tongpil; Kasahara, Hiroyuki; Bedgar, Diana L.; Youn, Buhyun; Lawrence, Paulraj K.; Gang, David R.; Halls, Steven C.; Park, HaJeung; Hilsenbeck, Jacqueline L.; Davin, Laurence B.; hide

    2003-01-01

    Despite the importance of plant lignans and isoflavonoids in human health protection (e.g. for both treatment and prevention of onset of various cancers) as well as in plant biology (e.g. in defense functions and in heartwood development), systematic studies on the enzymes involved in their biosynthesis have only recently begun. In this investigation, three NADPH-dependent aromatic alcohol reductases were comprehensively studied, namely pinoresinol-lariciresinol reductase (PLR), phenylcoumaran benzylic ether reductase (PCBER), and isoflavone reductase (IFR), which are involved in central steps to the various important bioactive lignans and isoflavonoids. Of particular interest was in determining how differing regio- and enantiospecificities are achieved with the different enzymes, despite each apparently going through similar enone intermediates. Initially, the three-dimensional x-ray crystal structures of both PLR_Tp1 and PCBER_Pt1 were solved and refined to 2.5 and 2.2 A resolutions, respectively. Not only do they share high gene sequence similarity, but their structures are similar, having a continuous alpha/beta NADPH-binding domain and a smaller substrate-binding domain. IFR (whose crystal structure is not yet obtained) was also compared (modeled) with PLR and PCBER and was deduced to have the same overall basic structure. The basis for the distinct enantio-specific and regio-specific reactions of PCBER, PLR, and IFR, as well as the reaction mechanism and participating residues involved (as identified by site-directed mutagenesis), are discussed.

  7. Identification and characterization of 2-naphthoyl-coenzyme A reductase, the prototype of a novel class of dearomatizing reductases.

    PubMed

    Eberlein, Christian; Estelmann, Sebastian; Seifert, Jana; von Bergen, Martin; Müller, Michael; Meckenstock, Rainer U; Boll, Matthias

    2013-06-01

    The enzymatic dearomatization of aromatic ring systems by reduction represents a highly challenging redox reaction in biology and plays a key role in the degradation of aromatic compounds under anoxic conditions. In anaerobic bacteria, most monocyclic aromatic growth substrates are converted to benzoyl-coenzyme A (CoA), which is then dearomatized to a conjugated dienoyl-CoA by ATP-dependent or -independent benzoyl-CoA reductases. It was unresolved whether or not related enzymes are involved in the anaerobic degradation of environmentally relevant polycyclic aromatic hydrocarbons (PAHs). In this work, a previously unknown dearomatizing 2-naphthoyl-CoA reductase was purified from extracts of the naphthalene-degrading, sulphidogenic enrichment culture N47. The oxygen-tolerant enzyme dearomatized the non-activated ring of 2-naphthoyl-CoA by a four-electron reduction to 5,6,7,8-tetrahydro-2-naphthoyl-CoA. The dimeric 150 kDa enzyme complex was composed of a 72 kDa subunit showing sequence similarity to members of the flavin-containing 'old yellow enzyme' family. NCR contained FAD, FMN, and an iron-sulphur cluster as cofactors. Extracts of Escherichia coli expressing the encoding gene catalysed 2-naphthoyl-CoA reduction. The identified NCR is a prototypical enzyme of a previously unknown class of dearomatizing arylcarboxyl-CoA reductases that are involved in anaerobic PAH degradation; it fundamentally differs from known benzoyl-CoA reductases.

  8. Crystal structures of pinoresinol-lariciresinol and phenylcoumaran benzylic ether reductases and their relationship to isoflavone reductases

    NASA Technical Reports Server (NTRS)

    Min, Tongpil; Kasahara, Hiroyuki; Bedgar, Diana L.; Youn, Buhyun; Lawrence, Paulraj K.; Gang, David R.; Halls, Steven C.; Park, HaJeung; Hilsenbeck, Jacqueline L.; Davin, Laurence B.; Lewis, Norman G.; Kang, ChulHee

    2003-01-01

    Despite the importance of plant lignans and isoflavonoids in human health protection (e.g. for both treatment and prevention of onset of various cancers) as well as in plant biology (e.g. in defense functions and in heartwood development), systematic studies on the enzymes involved in their biosynthesis have only recently begun. In this investigation, three NADPH-dependent aromatic alcohol reductases were comprehensively studied, namely pinoresinol-lariciresinol reductase (PLR), phenylcoumaran benzylic ether reductase (PCBER), and isoflavone reductase (IFR), which are involved in central steps to the various important bioactive lignans and isoflavonoids. Of particular interest was in determining how differing regio- and enantiospecificities are achieved with the different enzymes, despite each apparently going through similar enone intermediates. Initially, the three-dimensional x-ray crystal structures of both PLR_Tp1 and PCBER_Pt1 were solved and refined to 2.5 and 2.2 A resolutions, respectively. Not only do they share high gene sequence similarity, but their structures are similar, having a continuous alpha/beta NADPH-binding domain and a smaller substrate-binding domain. IFR (whose crystal structure is not yet obtained) was also compared (modeled) with PLR and PCBER and was deduced to have the same overall basic structure. The basis for the distinct enantio-specific and regio-specific reactions of PCBER, PLR, and IFR, as well as the reaction mechanism and participating residues involved (as identified by site-directed mutagenesis), are discussed.

  9. Immunological approach to the regulation of nitrate reductase in Monoraphidium braunii.

    PubMed

    Díez, J; López-Ruiz, A

    1989-02-01

    The effects of different culture conditions on nitrate reductase activity and nitrate reductase protein from Monoraphidium braunii have been studied, using two different immunological techniques, rocket immunoelectrophoresis and an enzyme-linked immunosorbent assay, to determine nitrate reductase protein. The nitrogen sources ammonium and glutamine repressed nitrate reductase synthesis, while nitrite, alanine, and glutamate acted as derepressors. There was a four- to eightfold increase of nitrate reductase activity and a twofold increase of nitrate reductase protein under conditions of nitrogen starvation versus growth on nitrate. Nitrate reductase synthesis was repressed in darkness. However, when Monoraphidium was grown under heterotrophic conditions with glucose as the carbon and energy source, the synthesis of nitrate reductase was maintained. With ammonium or darkness, changes in nitrate reductase activity correlated fairly well with changes in nitrate reductase protein, indicating that in both cases loss of activity was due to repression and not to inactivation of the enzyme. Experiments using methionine sulfoximine, to inhibit ammonium assimilation, showed that ammonium per se and not a product of its metabolism was the corepressor of the enzyme. The appearance of nitrate reductase activity after transferring the cells to induction media was prevented by cycloheximide and by 6-methylpurine, although in this latter case the effect was observed only in cells preincubated with the inhibitor for 1 h before the induction period.

  10. Recominant Pinoresino-Lariciresinol Reductase, Recombinant Dirigent Protein And Methods Of Use

    DOEpatents

    Lewis, Norman G.; Davin, Laurence B.; Dinkova-Kostova, Albena T.; Fujita, Masayuki , Gang; David R. , Sarkanen; Simo , Ford; Joshua D.

    2003-10-21

    Dirigent proteins and pinoresinol/lariciresinol reductases have been isolated, together with cDNAs encoding dirigent proteins and pinoresinol/lariciresinol reductases. Accordingly, isolated DNA sequences are provided from source species Forsythia intermedia, Thuja plicata, Tsuga heterophylla, Eucommia ulmoides, Linum usitatissimum, and Schisandra chinensis, which code for the expression of dirigent proteins and pinoresinol/lariciresinol reductases. In other aspects, replicable recombinant cloning vehicles are provided which code for dirigent proteins or pinoresinol/lariciresinol reductases or for a base sequence sufficiently complementary to at least a portion of dirigent protein or pinoresinol/lariciresinol reductase DNA or RNA to enable hybridization therewith. In yet other aspects, modified host cells are provided that have been transformed, transfected, infected and/or injected with a recombinant cloning vehicle and/or DNA sequence encoding dirigent protein or pinoresinol/lariciresinol reductase. Thus, systems and methods are provided for the recombinant expression of dirigent proteins and/or pinoresinol/lariciresinol reductases.

  11. Recombinant pinoresinol/lariciresinol reductase, recombinant dirigent protein, and methods of use

    DOEpatents

    Lewis, Norman G.; Davin, Laurence B.; Dinkova-Kostova, Albena T.; Fujita, Masayuki; Gang, David R.; Sarkanen, Simo; Ford, Joshua D.

    2001-04-03

    Dirigent proteins and pinoresinol/lariciresinol reductases have been isolated, together with cDNAs encoding dirigent proteins and pinoresinol/lariciresinol reductases. Accordingly, isolated DNA sequences are provided which code for the expression of dirigent proteins and pinoresinol/lariciresinol reductases. In other aspects, replicable recombinant cloning vehicles are provided which code for dirigent proteins or pinoresinol/lariciresinol reductases or for a base sequence sufficiently complementary to at least a portion of dirigent protein or pinoresinol/lariciresinol reductase DNA or RNA to enable hybridization therewith. In yet other aspects, modified host cells are provided that have been transformed, transfected, infected and/or injected with a recombinant cloning vehicle and/or DNA sequence encoding dirigent protein or pinoresinol/lariciresinol reductase. Thus, systems and methods are provided for the recombinant expression of dirigent proteins and/or pinoresinol/lariciresinol reductases.

  12. Mercuric chloride-induced gastrin/cholecystokinin 8 immunoreactivity in the central nervous system of the terrestrial slug Semperula maculata: an immunohistochemical study.

    PubMed

    Londhe, Sunil; Kamble, Nitin

    2013-12-01

    We measured the immunoreactivity of the neuropeptide gastrin cholecystokinin 8 (gastrin/CCK 8) in neurons of the terrestrial slug Semperula maculata following acute treatment with mercuric chloride (HgCl2). The distribution of gastrin/CCK 8 was analyzed in neurons of different regions, specifically from cerebral ganglia (procerebrum (pro-c), mesocerebrum (meso-c) and metacerebrum (meta-c). In the control group, neurons of pedal, pleural, parietal and visceral ganglia showed positive immunoreactivity using vertebrate antiserum against gastrin/CCK 8. Gastrin/CCK 8 immunoreactivity was also seen in the fibers and neuropil region of all ganglia. In the cerebral ganglion, 10, 12 and 8 % of the neurons from pro-c, meso-c and meta-c, respectively, were stained with the antibody. The immunostaining was increased in neurons (giant, large, medium and small) after HgCl2 treatment. The treatment greatly increased the mucin content within the neurons. Exposure to HgCl2 enhanced gastrin immunoreactivity in the neurons and this increased with time. Results are discussed in the context of neuropathology in cerebral ganglia associated with the feeding behavior of Semperula maculata.

  13. Regulation of Sirt1/Nrf2/TNF-α signaling pathway by luteolin is critical to attenuate acute mercuric chloride exposure induced hepatotoxicity

    PubMed Central

    Yang, Daqian; Tan, Xiao; Lv, Zhanjun; Liu, Biying; Baiyun, Ruiqi; Lu, Jingjing; Zhang, Zhigang

    2016-01-01

    Inorganic mercury, though a key component of pediatric vaccines, is an environmental toxicant threatening human health via accumulating oxidative stress in part. Luteolin has been of great interest because of its antiinflammatory, anticarcinogenic and antioxidative effects. Here we hypothesized that luteolin would attenuate hepatotoxicity induced by acute inorganic mercury exposure. Kunming mice were treated with luteolin (100 mg/kg) 24 h after administration of 4 mg/kg mercuric chloride (HgCl2). The results showed that luteolin ameliorated HgCl2 induced anemia and hepatotoxicity, regulating radical oxygen species (ROS) production and hepatocyte viability in vitro and oxidative stress and apoptosis in vivo. Furthermore, luteolin reversed the changes in levels of inflammation- and apoptosis-related proteins involving NF-κB, TNF-α, Sirt1, mTOR, Bax, p53, and Bcl-2, and inhibited p38 MAPK activation. Luteolin enhanced antioxidant defense system based on Keap1, Nrf2, HO-1, NQO1, and KLF9. Moreover, luteolin did not affect miRNA-146a expression. Collectively, our findings, for the first time, elucidate a precise mechanism for attenuation of HgCl2-induced liver dysfunction by dietary luteolin via regulating Sirt1/Nrf2/TNF-α signaling pathway, and provide a foundation for further study of luteolin as a novel therapeutic agent against inorganic mercury poisoning. PMID:27853236

  14. Optical, electrical and surface characterization of mercuric iodide platelets grown in the HgI{sub 2}-HI-H{sub 2}O system

    SciTech Connect

    Fornaro, L.; Chen, H.; Chattopadhyay, K.; Chen, K.T.; Burger, A.

    1998-12-31

    The optical, electrical and surface properties of mercuric iodide platelets grown from solution in a HgI{sub 2}-HI-H{sub 2}O system were investigated by comparing them with Physical Vapor Transport (PVT) grown crystals. The absence of bulk imperfections and the uniformity of the as-grown surfaces and the KI solution etched surfaces were confirmed by optical microscopy. The as-grown surface uniformity is higher for solution grown crystals than that of PVT crystals, since the platelets do not have to be cleaved or polished. AFM studies show that the roughness for the cleaved, aged and etched surfaces were 0.06 nm, 0.48 nm and 0.3 nm respectively. Low temperature photoluminescence properties were measured for the two kinds of crystals and will be discussed. However, I-V curves give higher current density and lower apparent resistivity values for the solution grown than for PVT grown crystals. Correlations between optical and surface quality as well as the electrical properties of the crystals grown from both solution and PVT methods are also discussed.

  15. Curcuma longa Linn. extract and curcumin protect CYP 2E1 enzymatic activity against mercuric chloride-induced hepatotoxicity and oxidative stress: A protective approach.

    PubMed

    Joshi, Deepmala; Mittal, Deepak Kumar; Shukla, Sangeeta; Srivastav, Sunil Kumar; Dixit, Vaibhav A

    2017-03-20

    The present investigation has been conducted to evaluate the therapeutic potential of Curcuma longa (200mgkg(-1), po) and curcumin (80mgkg(-1), po) for their hepatoprotective efficacy against mercuric chloride (HgCl2: 12μmolkg(-1), ip; once only) hepatotoxicity. The HgCl2 administration altered various biochemical parameters, including transaminases, alkaline phosphatase, lactate dehydrogenase, bilirubin, gamma-glutamyl transferase, triglycerides and cholesterol contents with a concomitant decline in protein and albumin concentration in serum which were restored towards control by therapy of Curcuma longa or curcumin. On the other hand, both treatments showed a protective effect on drug metabolizing enzymes viz. aniline hydroxylase (AH) and amidopyrine-N-demethylase (AND), hexobarbitone induced sleep time and BSP retention. Choleretic, 1,1-diphenyl-2-picryl-hydrazil (DPPH)-free radical scavenging activities and histological studies also supported the biochemical findings. The present study concludes that Curcuma longa extract or curcumin has the ability to alleviate the hepatotoxic effects caused by HgCl2 in rats.

  16. Spectroscopic studies of the copper sites in wild-type pseudomonas stutzeri N sub 2 O reductase and in an inactive protein isolated from a mutant deficient in copper-site biosynthesis

    SciTech Connect

    Dooley, D.M.; McGuirl, M.A.; Rosenzweig, A.C.; Landin, J.A. ); Scott, R.A. ); Zumft, W.G. ); Devlin, F.; Stephens, P.J. )

    1991-07-24

    The results of studies of circular dichroism, magnetic circular dichroism, and x-ray absorption and fluorescence spectroscopy used to further characterize the copper sites in native N{sub 2}O reductase and in a mutant protein isolated from a strain deficient in the biosynthesis of the N{sub 2}O reductase copper sites that contains only 2 copper ions/protein molecule, are reported herein. Both magnetic circular dichroism and x-ray absorption (Cu K-edge and EXADS) data are consistent with the presence of (on average) one Cu{sub A}-type site per protein in the mutant N{sub 2}O reductase. Comparisons of the near-infrared circular dichroism spectra of the oxidized native and mutant N{sub 2}O reductases suggest that transitions at 7,200 and 9,500 cm{sup {minus}1} in the native enzyme are associated with copper sites other than the Cu{sub A}-type sites. These are the first electronic spectral features that can be attributed to non-Cu-type sites in N{sub 2}O reductase. The near-infrared bands are believed to be significantly less intense in preparations of N{sub 2}O reductase that display lower specific activities. Several electronic transitions are resolved in the circular and magnetic circular dichroism spectra of dithionite-reduced N{sub 2}O reductase. Notably a near-infrared band is observed in the circular dichroism spectrum at 8,200 cm{sup {minus}1}. Theories are presented to explain the reported data. 29 refs., 8 figs., 1 tab.

  17. The X-ray crystal structure of APR-B, an atypical adenosine 5'-phosphosulfate reductase from Physcomitrella patens.

    PubMed

    Stevenson, Clare E M; Hughes, Richard K; McManus, Michael T; Lawson, David M; Kopriva, Stanislav

    2013-11-15

    Sulfonucleotide reductases catalyse the first reductive step of sulfate assimilation. Their substrate specificities generally correlate with the requirement for a [Fe4S4] cluster, where adenosine 5'-phosphosulfate (APS) reductases possess a cluster and 3'-phosphoadenosine 5'-phosphosulfate reductases do not. The exception is the APR-B isoform of APS reductase from the moss Physcomitrella patens, which lacks a cluster. The crystal structure of APR-B, the first for a plant sulfonucleotide reductase, is consistent with a preference for APS. Structural conservation with bacterial APS reductase rules out a structural role for the cluster, but supports the contention that it enhances the activity of conventional APS reductases.

  18. Polyethylene glycol conjugation enhances the nitrite reductase activity of native and cross-linked hemoglobin.

    PubMed

    Lui, Francine E; Dong, Pengcheng; Kluger, Ronald

    2008-10-07

    Although stabilized hemoglobins have been evaluated as oxygen-carrying replacements for red cells in transfusions, in vivo evaluations have noted that these materials are associated with vasoactivity, a serious complication. Scavenging of endogenous nitric oxide by the deoxyheme sites of the stabilized proteins is one likely source of vasoactivity. Recent reports indicate that modification of cell-free hemoglobin derivatives with multiple chains of polyethylene glycol (PEG) suppresses vasoactivity. Gladwin and co-workers observed that the nitrite reductase activity of hemoglobin serves as a major endogenous source of nitric oxide. If PEG conjugation leads to enhanced nitrite reductase activity, this could compensate for scavenged endogenous nitric oxide. To test this possibility, the rates of conversion of nitrite ion to nitric oxide by altered hemoglobins with and without PEG were measured at 25 degrees C. Fumaryl (alpha99-alpha99) cross-linked hemoglobin reacts with nitrite with a bimolecular rate constant of 0.52 M (-1) s (-1), which is comparable to that associated with native hemoglobin (0.25 M (-1) s (-1)). Addition of PEG chains to the cross-linked hemoglobin at beta-Cys93 (alphaalpha-Hb-PEG5K 2) results in a material that produces nitric oxide much more rapidly ( k = 1.41 M (-1) s (-1)). R-State-stabilized hemoglobins with multiple PEG chains (Hb-PEG5K 2 and Hb-PEG5K 6) react 10 times faster with nitrite to produce nitric oxide than does native hemoglobin ( k = 2.5 and 2.4 M (-1) s (-1), respectively). These results, showing enhanced production of nitric oxide resulting from an increased proportion of the protein residing in the R-state, are consistent with the decrease in vasoactivity associated with PEG conjugation.

  19. Antimicrobial activity and physical characterization of silver nanoparticles green synthesized using nitrate reductase from Fusarium oxysporum.

    PubMed

    Gholami-Shabani, Mohammadhassan; Akbarzadeh, Azim; Norouzian, Dariush; Amini, Abdolhossein; Gholami-Shabani, Zeynab; Imani, Afshin; Chiani, Mohsen; Riazi, Gholamhossein; Shams-Ghahfarokhi, Masoomeh; Razzaghi-Abyaneh, Mehdi

    2014-04-01

    Nanostructures from natural sources have received major attention due to wide array of biological activities and less toxicity for humans, animals, and the environment. In the present study, silver nanoparticles were successfully synthesized using a fungal nitrate reductase, and their biological activity was assessed against human pathogenic fungi and bacteria. The enzyme was isolated from Fusarium oxysporum IRAN 31C after culturing on malt extract-glucose-yeast extract-peptone (MGYP) medium. The enzyme was purified by a combination of ultrafiltration and ion exchange chromatography on DEAE Sephadex and its molecular weight was estimated by gel filtration on Sephacryl S-300. The purified enzyme had a maximum yield of 50.84 % with a final purification of 70 folds. With a molecular weight of 214 KDa, it is composed of three subunits of 125, 60, and 25 KDa. The purified enzyme was successfully used for synthesis of silver nanoparticles in a way dependent upon NADPH using gelatin as a capping agent. The synthesized silver nanoparticles were characterized by X-ray diffraction, dynamic light scattering spectroscopy, and transmission and scanning electron microscopy. These stable nonaggregating nanoparticles were spherical in shape with an average size of 50 nm and a zeta potential of -34.3. Evaluation of the antimicrobial effects of synthesized nanoparticles by disk diffusion method showed strong growth inhibitory activity against all tested human pathogenic fungi and bacteria as evident from inhibition zones that ranged from 14 to 25 mm. Successful green synthesis of biologically active silver nanoparticles by a nitrate reductase from F. oxysporum in the present work not only reduces laborious downstream steps such as purification of nanoparticle from interfering cellular components, but also provides a constant source of safe biologically-active nanomaterials with potential application in agriculture and medicine.

  20. The respiratory arsenate reductase from Bacillus selenitireducens strain MLS10

    USGS Publications Warehouse

    Afkar, E.; Lisak, J.; Saltikov, C.; Basu, P.; Oremland, R.S.; Stolz, J.F.

    2003-01-01

    The respiratory arsenate reductase from the Gram-positive, haloalkaliphile, Bacillus selenitireducens strain MLS10 was purified and characterized. It is a membrane bound heterodimer (150 kDa) composed of two subunits ArrA (110 kDa) and ArrB (34 kDa), with an apparent Km for arsenate of 34 ??M and Vmax of 2.5 ??mol min-1 mg-1. Optimal activity occurred at pH 9.5 and 150 g l-1 of NaCl. Metal analysis (inductively coupled plasma mass spectrometry) of the holoenzyme and sequence analysis of the catalytic subunit (ArrA; the gene for which was cloned and sequenced) indicate it is a member of the DMSO reductase family of molybdoproteins. ?? 2003 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

  1. Ranirestat as a therapeutic aldose reductase inhibitor for diabetic complications.

    PubMed

    Giannoukakis, Nick

    2008-04-01

    There are currently very few drugs available to directly treat diabetic complications. Those that are indicated clinically provide symptomatic relief and do not address the underlying biochemical problems. The involvement of the sorbitol pathway in complications has provided mechanistic insights into the biochemistry of complications and the key enzyme, aldose reductase, has become an attractive pharmacologic target. Among the aldose reductase inhibitors, the most promising is ranirestat. This review outlines the studies with ranirestat and compares its efficacy with other similar inhibitors. A survey of in vitro and in vivo studies was conducted, and with publicly available data from clinical trials, ranirestat efficacy was compared with other similar agents. Ranirestat is safe, exhibits some efficacy and is perhaps the only agent advanced enough in clinical trials to warrant further consideration for diabetic complications.

  2. Pyrroline-5-Carboxylate Reductase in Soybean Nodules 1

    PubMed Central

    Chilson, Oscar P.; Kelly-Chilson, Anne E.; Schneider, Julie D.

    1992-01-01

    Characteristics of pyrroline-5-carboxylate reductase (P5CR) from Bradyrhizobium japonicum bacteroids and cultured rhizobia were compared with those of the enzyme in soybean nodule host cytosol. Reductase from host cytosol differed from that in bacteroids in: (a) the effect of pH on enzymic activity, (b) the capacity to catalyze both reduction of pyrroline-5-carboxylic acid and NAD+-dependent proline oxidation, (c) apparent affinities for pyrroline-5-carboxylic acid, and (d) sensitivities to inhibition by NADP+ and proline. The K1 for proline inhibition of P5CR in bacteroid cytosol was 1.8 millimolar. The properties of P5CR in B. japonicum and bacteroid cytosol were similar. The specific activities of P5CR in the cytosolic fractions of the nodule host and the bacteroid compartment were also comparable. PMID:16668837

  3. Characterization of 12-Oxo-Phytodienoic Acid Reductase in Corn

    PubMed Central

    Vick, Brady A.; Zimmerman, Don C.

    1986-01-01

    12-Oxo-phytodienoic acid reductase, an enzyme of the biosynthetic pathway that converts linolenic acid to jasmonic acid, has been characterized from the kernel and seedlings of corn (Zea mays L.). The molecular weight of the enzyme, estimated by gel filtration, was 54,000. Optimum enzyme activity was observed over a broad pH range, from pH 6.8 to 9.0. The enzyme had a Km of 190 micromolar for its substrate, 12-oxo-phytodienoic acid. The preferred reductant was NADPH, for which the enzyme exhibited a Km of 13 micromolar, compared with 4.2 millimolar for NADH. Reductase activity was low in the corn kernel but increased five-fold by the fifth day after germination and then gradually declined. PMID:16664582

  4. [Properties of a nitrite reductase inhibitor protein from Pseudomonas aeruginosa].

    PubMed

    Karapetian, A V; Nalbandian, R M

    1993-08-01

    The amino acid composition and major physico-chemical properties of the "nonblue" copper protein isolated earlier from Pseudomonas aeruginosa have been determined. It has been found that the azurin oxidase, cytochrome c551 oxidase and superoxide dismutase activities of the enzyme are inhibited by this protein. The inhibition seems to be due to the protein interaction with the electron-accepting center of nitrite reductase.

  5. Differential expression of disulfide reductase enzymes in a free-living platyhelminth (Dugesia dorotocephala)

    PubMed Central

    Herrera-Juárez, Álvaro Miguel; Martínez-González, José de Jesús; del Arenal Mena, Irene Patricia; Flores-Herrera, Óscar

    2017-01-01

    A search of the disulfide reductase activities expressed in the adult stage of the free-living platyhelminth Dugesia dorotocephala was carried out. Using GSSG or DTNB as substrates, it was possible to obtain a purified fraction containing both GSSG and DTNB reductase activities. Through the purification procedure, both disulfide reductase activities were obtained in the same chromatographic peak. By mass spectrometry analysis of peptide fragments obtained after tryptic digestion of the purified fraction, the presence of glutathione reductase (GR), thioredoxin-glutathione reductase (TGR), and a putative thioredoxin reductase (TrxR) was detected. Using the gold compound auranofin to selectively inhibit the GSSG reductase activity of TGR, it was found that barely 5% of the total GR activity in the D. dorotocephala extract can be assigned to GR. Such strategy did allow us to determine the kinetic parameters for both GR and TGR. Although It was not possible to discriminate DTNB reductase activity due to TrxR from that of TGR, a chromatofocusing experiment with a D. dorotocephala extract resulted in the obtention of a minor protein fraction enriched in TrxR, strongly suggesting its presence as a functional protein. Thus, unlike its parasitic counterparts, in the free-living platyhelminth lineage the three disulfide reductases are present as functional proteins, albeit TGR is still the major disulfide reductase involved in the reduction of both Trx and GSSG. This fact suggests the development of TGR in parasitic flatworms was not linked to a parasitic mode of life. PMID:28787021

  6. Aldose and aldehyde reductases : structure-function studies on the coenzyme and inhibitor-binding sites.

    SciTech Connect

    El-Kabbani, O.; Old, S. E.; Ginell, S. L.; Carper, D. A.; Biosciences Division; Monash Univ.; NIH

    1999-09-03

    PURPOSE: To identify the structural features responsible for the differences in coenzyme and inhibitor specificities of aldose and aldehyde reductases. METHODS: The crystal structure of porcine aldehyde reductase in complex with NADPH and the aldose reductase inhibitor sorbinil was determined. The contribution of each amino acid lining the coenzyme-binding site to the binding of NADPH was calculated using the Discover package. In human aldose reductase, the role of the non-conserved Pro 216 (Ser in aldehyde reductase) in the binding of coenzyme was examined by site-directed mutagenesis. RESULTS: Sorbinil binds to the active site of aldehyde reductase and is hydrogen-bonded to Trp 22, Tyr 50, His 113, and the non-conserved Arg 312. Unlike tolrestat, the binding of sorbinil does not induce a change in the side chain conformation of Arg 312. Mutation of Pro 216 to Ser in aldose reductase makes the binding of coenzyme more similar to that of aldehyde reductase. CONCLUSIONS: The participation of non-conserved active site residues in the binding of inhibitors and the differences in the structural changes required for the binding to occur are responsible for the differences in the potency of inhibition of aldose and aldehyde reductases. We report that the non-conserved Pro 216 in aldose reductase contributes to the tight binding of NADPH.

  7. Autoregulation of the Synthesis of Nitrate Reductase in Aspergillus nidulans

    PubMed Central

    Cove, D. J.; Pateman, J. A.

    1969-01-01

    In Aspergillus nidulans, the syntheses of nitrate and nitrite reductases are induced by nitrate, and are repressed by ammonium. It is possible in wild-type strains to overcome partially the repressive effect of ammonium, by the addition of high concentrations of nitrate to the growth medium. Mutations which lead to the production of abnormal nitrate reductase affect in addition the control of the synthesis of the nitrate-metabolizing enzymes, which in these strains are produced constitutively. That this is not due to the accumulation of an internal inducer has now been shown, as these mutants have been found to be unable to respond to nitrate induction in the presence of ammonium in the same way as do wild-type strains. To explain these findings, we propose that the nitrate reductase molecule provides the recognition site for nitrate in the control system, such that when it is not complexed with nitrate it acts as a co-repressor, and, when it is complexed, as a co-inducer. PMID:5776531

  8. The effect of quercetin and galangin on glutathione reductase.

    PubMed

    Paulíková, Helena; Berczeliová, Elena

    2005-12-01

    Quercetin and galangin can change the activity of glutathione reductase. Quercetin (a catechol structure in the B-ring) and galangin (any hydroxyl group in the B-ring) have different biological activities but, both possess high antioxidant abilities. Quercetin during the antioxidative action, is converted into an oxidized products (o-semiquinone and o-quinone), and subsequently glutathionyl adducts may be formed or SH-enzyme can be inhibited. We have tried to see whether inhibition of glutathione reductase (GR) can be influenced by preincubation of enzyme with NADPH (a creation of reduced form of enzyme, GRH(2)) and whether diaphorase activity of the enzyme is decreased by these flavonoids. The results confirmed that quercetin inhibits GRH(2) and inhibition is reduced by addition of EDTA or N-acetylcysteine. Both of flavonoids have no effect on diaphorase activity of glutathione reductase and this enzyme could increase the production of free radicals by catalysis of reduction of o-quinone during action of quercetin in vivo.

  9. Structural and functional diversity of ferredoxin-NADP(+) reductases.

    PubMed

    Aliverti, Alessandro; Pandini, Vittorio; Pennati, Andrea; de Rosa, Matteo; Zanetti, Giuliana

    2008-06-15

    Although all ferredoxin-NADP(+) reductases (FNRs) catalyze the same reaction, i.e. the transfer of reducing equivalents between NADP(H) and ferredoxin, they belong to two unrelated families of proteins: the plant-type and the glutathione reductase-type of FNRs. Aim of this review is to provide a general classification scheme for these enzymes, to be used as a framework for the comparison of their properties. Furthermore, we report on some recent findings, which significantly increased the understanding of the structure-function relationships of FNRs, i.e. the ability of adrenodoxin reductase and its homologs to catalyze the oxidation of NADP(+) to its 4-oxo derivative, and the properties of plant-type FNRs from non-photosynthetic organisms. Plant-type FNRs from bacteria and Apicomplexan parasites provide examples of novel ways of FAD- and NADP(H)-binding. The recent characterization of an FNR from Plasmodium falciparum brings these enzymes into the field of drug design.

  10. Rat liver thioredoxin and thioredoxin reductase: purification and characterization.

    PubMed

    Luthman, M; Holmgren, A

    1982-12-21

    A reproducible scheme has been developed for the preparation of rat liver thioredoxin and thioredoxin reductase (EC 1.6.4.5) by using assays based on reduction of insulin and 5,5'-dithiobis(2-nitrobenzoic acid), respectively. Both proteins were purified to homogeneity, as judged from polyacrylamide gel electrophoresis. Thioredoxin had a molecular weight of 12 000 and contained about 110 amino acids including 4 half-cystines and an NH2-terminal valine. Peptide maps of reduced and carboxymethylated thioredoxin showed that the protein had the active center sequence -Cys-Gly-Pro-Cys-Lys-Met- characteristic of thioredoxins also from procaryotes. Prolonged air oxidation of fully reduced thioredoxin created inactive, aggregated disulfide-containing molecules. Thioredoxin reductase showed a subunit molecular weight of 58 000 and a native molecular weight of 116 000. The enzyme was highly specific for NADPH with a Km of 6 microM. It contained FAD as prosthetic group and was sensitive to inhibition by arsenite. Thioredoxin reductase had a Km of 2.5 microM for rat and calf liver thioredoxin and a Kcat of 3000 min-1.

  11. Structures of carboxylic acid reductase reveal domain dynamics underlying catalysis.

    PubMed

    Gahloth, Deepankar; Dunstan, Mark S; Quaglia, Daniela; Klumbys, Evaldas; Lockhart-Cairns, Michael P; Hill, Andrew M; Derrington, Sasha R; Scrutton, Nigel S; Turner, Nicholas J; Leys, David

    2017-09-01

    Carboxylic acid reductase (CAR) catalyzes the ATP- and NADPH-dependent reduction of carboxylic acids to the corresponding aldehydes. The enzyme is related to the nonribosomal peptide synthetases, consisting of an adenylation domain fused via a peptidyl carrier protein (PCP) to a reductase termination domain. Crystal structures of the CAR adenylation-PCP didomain demonstrate that large-scale domain motions occur between the adenylation and thiolation states. Crystal structures of the PCP-reductase didomain reveal that phosphopantetheine binding alters the orientation of a key Asp, resulting in a productive orientation of the bound nicotinamide. This ensures that further reduction of the aldehyde product does not occur. Combining crystallography with small-angle X-ray scattering (SAXS), we propose that molecular interactions between initiation and termination domains are limited to competing PCP docking sites. This theory is supported by the fact that (R)-pantetheine can support CAR activity for mixtures of the isolated domains. Our model suggests directions for further development of CAR as a biocatalyst.

  12. Early diagnosis and management of 5 alpha-reductase deficiency.

    PubMed Central

    Odame, I; Donaldson, M D; Wallace, A M; Cochran, W; Smith, P J

    1992-01-01

    Two siblings of Pakistani origin, karyotype 46 XY, were born with predominantly female external genitalia with minute phallus, bifid scrotum, urogenital sinus, and palpable gonads. The older sibling at the age of 8 days showed an adequate testosterone response to human chorionic gonadotrophin (hCG) stimulation. The diagnosis of 5 alpha-reductase deficiency was made at age 6 years when no 5 alpha-reduced glucocorticoid metabolites were detectable in urine even after tetracosactrin (Synacthen) stimulation. In the younger sibling the diagnosis of 5 alpha-reductase deficiency was provisionally made at the early age of 3 days on the basis of high urinary tetrahydrocortisol (THF)/allotetrahydrocortisol (5 alpha-THF) ratio and this ratio increased with age confirming the diagnosis. Plasma testosterone: dihydrotestosterone (DHT) ratio before and after hCG stimulation was within normal limits at age 3 days but was raised at age 9 months. Topical DHT cream application to the external genitalia promoted significant phallic growth in both siblings and in the older sibling corrective surgery was facilitated. In prepubertal male pseudohermaphrodites with normal or raised testosterone concentrations, phallic growth in response to DHT cream treatment could be an indirect confirmation of 5 alpha-reductase deficiency. Images Figure 1 PMID:1626992

  13. Phosphoglycerate kinase acts in tumour angiogenesis as a disulphide reductase

    NASA Astrophysics Data System (ADS)

    Lay, Angelina J.; Jiang, Xing-Mai; Kisker, Oliver; Flynn, Evelyn; Underwood, Anne; Condron, Rosemary; Hogg, Philip J.

    2000-12-01

    Disulphide bonds in secreted proteins are considered to be inert because of the oxidizing nature of the extracellular milieu. An exception to this rule is a reductase secreted by tumour cells that reduces disulphide bonds in the serine proteinase plasmin. Reduction of plasmin initiates proteolytic cleavage in the kringle 5 domain and release of the tumour blood vessel inhibitor angiostatin. New blood vessel formation or angiogenesis is critical for tumour expansion and metastasis. Here we show that the plasmin reductase isolated from conditioned medium of fibrosarcoma cells is the glycolytic enzyme phosphoglycerate kinase. Recombinant phosphoglycerate kinase had the same specific activity as the fibrosarcoma-derived protein. Plasma of mice bearing fibrosarcoma tumours contained several-fold more phosphoglycerate kinase, as compared with mice without tumours. Administration of phosphoglycerate kinase to tumour-bearing mice caused an increase in plasma levels of angiostatin, and a decrease in tumour vascularity and rate of tumour growth. Our findings indicate that phosphoglycerate kinase not only functions in glycolysis but is secreted by tumour cells and participates in the angiogenic process as a disulphide reductase.

  14. Aldo-Keto Reductases 1B in Adrenal Cortex Physiology

    PubMed Central

    Pastel, Emilie; Pointud, Jean-Christophe; Martinez, Antoine; Lefrançois-Martinez, A. Marie

    2016-01-01

    Aldose reductase (AKR1B) proteins are monomeric enzymes, belonging to the aldo-keto reductase (AKR) superfamily. They perform oxidoreduction of carbonyl groups from a wide variety of substrates, such as aliphatic and aromatic aldehydes or ketones. Due to the involvement of human aldose reductases in pathologies, such as diabetic complications and cancer, AKR1B subgroup enzymatic properties have been extensively characterized. However, the issue of AKR1B function in non-pathologic conditions remains poorly resolved. Adrenal activities generated large amount of harmful aldehydes from lipid peroxidation and steroidogenesis, including 4-hydroxynonenal (4-HNE) and isocaproaldehyde (4-methylpentanal), which can both be reduced by AKR1B proteins. More recently, some AKR1B isoforms have been shown to be endowed with prostaglandin F synthase (PGFS) activity, suggesting that, in addition to possible scavenger function, they could instigate paracrine signals. Interestingly, the adrenal gland is one of the major sites for human and murine AKR1B expression, suggesting that their detoxifying/signaling activity could be specifically required for the correct handling of adrenal function. Moreover, chronic effects of ACTH result in a coordinated regulation of genes encoding the steroidogenic enzymes and some AKR1B isoforms. This review presents the molecular mechanisms accounting for the adrenal-specific expression of some AKR1B genes. Using data from recent mouse genetic models, we will try to connect their enzymatic properties and regulation with adrenal functions. PMID:27499746

  15. Cloning and Sequence Analysis of Two Pseudomonas Flavoprotein Xenobiotic Reductases

    PubMed Central

    Blehert, David S.; Fox, Brian G.; Chambliss, Glenn H.

    1999-01-01

    The genes encoding flavin mononucleotide-containing oxidoreductases, designated xenobiotic reductases, from Pseudomonas putida II-B and P. fluorescens I-C that removed nitrite from nitroglycerin (NG) by cleavage of the nitroester bond were cloned, sequenced, and characterized. The P. putida gene, xenA, encodes a 39,702-Da monomeric, NAD(P)H-dependent flavoprotein that removes either the terminal or central nitro groups from NG and that reduces 2-cyclohexen-1-one but did not readily reduce 2,4,6-trinitrotoluene (TNT). The P. fluorescens gene, xenB, encodes a 37,441-Da monomeric, NAD(P)H-dependent flavoprotein that exhibits fivefold regioselectivity for removal of the central nitro group from NG and that transforms TNT but did not readily react with 2-cyclohexen-1-one. Heterologous expression of xenA and xenB was demonstrated in Escherichia coli DH5α. The transcription initiation sites of both xenA and xenB were identified by primer extension analysis. BLAST analyses conducted with the P. putida xenA and the P. fluorescens xenB sequences demonstrated that these genes are similar to several other bacterial genes that encode broad-specificity flavoprotein reductases. The prokaryotic flavoprotein reductases described herein likely shared a common ancestor with old yellow enzyme of yeast, a broad-specificity enzyme which may serve a detoxification role in antioxidant defense systems. PMID:10515912

  16. Quinone Reductase Induction as a Biomarker for Cancer Chemoprevention⊥

    PubMed Central

    Cuendet, Muriel; Oteham, Carol P.; Moon, Richard C.; Pezzuto, John M.

    2007-01-01

    Chemoprevention involves the use of natural or synthetic substances to reduce the risk of developing cancer. Strategies for protecting cells from initiation events include decreasing metabolic enzymes responsible for generating reactive species (phase I enzymes) while increasing phase II enzymes that can deactivate radicals and electrophiles known to intercede in normal cellular processes. Reduction of electrophilic quinones by quinone reductase is an important detoxification pathway. Following evaluation of approximately 3000 plant and marine organism extracts, the number characterized as “active” was established in the range of 12% of the total, and over 60 active compounds have been isolated as quinone reductase inducers. One of them, isoliquiritigenin (1), isolated from tonka bean, was shown to be a monofunctional inducer by having similar quinone reductase inducing ability in wild-type Hepa 1c1c7 cells and two mutant cell lines. To further investigate the mechanism of induction, HepG2 human hepatoma cells stably transfected with ARE-luciferase plasmid were used. Isoliquiritigenin (1) significantly induced the luciferase activity in a dose-dependent manner. On the basis of these results, a full-term cancer chemoprevention study was conducted with 7,12-dimethylbenz[a]anthracene (DMBA)-treated female Sprague-Dawley rats. Dietary administration of 1 increased tumor latency. Based on these promising preliminary results, additional mechanistic studies are underway, as well as full-term carcinogenesis studies with chronic administration schedules. PMID:16562858

  17. The existence and significance of a mitochondrial nitrite reductase.

    PubMed

    Nohl, Hans; Staniek, Katrin; Kozlov, Andrey V

    2005-01-01

    The physiological functions of nitric oxide (NO) are well established. The finding that the endothelium-derived relaxing factor (EDRF) is NO was totally unexpected. It was shown that NO is a reaction product of an enzymatically catalyzed, overall, 5-electron oxidation of guanidinium nitrogen from L-arginine followed by the release of the free radical species NO. NO is synthesized by a single protein complex supported by cofactors, coenzymes (such as tetrahydrobiopterin) and cytochrome P450. The latter can uncouple from substrate oxidation producing O2*- radicals. The research groups of Richter [Ghafourifar P, Richter C. Nitric oxide synthase activity in mitochondria. FEBS Lett 1997; 418: 291-296.] and Boveris [Giulivi C, Poderoso JJ, Boveris A. Production of nitric oxide by mitochondria. J Biol Chem 1998; 273: 11038-11043.] identified a mitochondrial NO synthase (NOS). There are, however, increasing reports demonstrating that mitochondrial NO is derived from cytosolic NOS belonging to the Ca2+-dependent enzymes. NO was thought to control cytochrome oxidase. This assumption is controversial due to the life-time of NO in biological systems (millisecond range). We found a nitrite reductase in mitochondria which is of major interest. Any increase of nitrite in the tissue which is the first oxidation product of NO, for instance following NO donors, will stimulate NO-recycling via mitochondrial nitrite reductase. In this paper, we describe the identity and the function of mitochondrial nitrite reductase and the consequences of NO-recycling in the metabolic compartment of mitochondria.

  18. Catalytic mechanism and substrate selectivity of aldo-keto reductases: insights from structure-function studies of Candida tenuis xylose reductase.

    PubMed

    Kratzer, Regina; Wilson, David K; Nidetzky, Bernd

    2006-09-01

    Aldo-keto reductases (AKRs) constitute a large protein superfamily of mainly NAD(P)-dependent oxidoreductases involved in carbonyl metabolism. Catalysis is promoted by a conserved tetrad of active site residues (Tyr, Lys, Asp and His). Recent results of structure-function relationship studies for xylose reductase (AKR2B5) require an update of the proposed catalytic mechanism. Electrostatic stabilization by the epsilon-NH3+ group of Lys is a key source of catalytic power of xylose reductase. A molecular-level analysis of the substrate binding pocket of xylose reductase provides a case of how a very broadly specific AKR achieves the requisite selectivity for its physiological substrate and could serve as the basis for the design of novel reductases with improved specificities for biocatalytic applications.

  19. Methionine Sulfoxide Reductases Are Essential for Virulence of Salmonella Typhimurium

    PubMed Central

    Rouf, Syed Fazle; Kitowski, Vera; Böhm, Oliver M.; Rhen, Mikael; Jäger, Timo; Bange, Franz-Christoph

    2011-01-01

    Production of reactive oxygen species represents a fundamental innate defense against microbes in a diversity of host organisms. Oxidative stress, amongst others, converts peptidyl and free methionine to a mixture of methionine-S- (Met-S-SO) and methionine-R-sulfoxides (Met-R-SO). To cope with such oxidative damage, methionine sulfoxide reductases MsrA and MsrB are known to reduce MetSOs, the former being specific for the S-form and the latter being specific for the R-form. However, at present the role of methionine sulfoxide reductases in the pathogenesis of intracellular bacterial pathogens has not been fully detailed. Here we show that deletion of msrA in the facultative intracellular pathogen Salmonella (S.) enterica serovar Typhimurium increased susceptibility to exogenous H2O2, and reduced bacterial replication inside activated macrophages, and in mice. In contrast, a ΔmsrB mutant showed the wild type phenotype. Recombinant MsrA was active against free and peptidyl Met-S-SO, whereas recombinant MsrB was only weakly active and specific for peptidyl Met-R-SO. This raised the question of whether an additional Met-R-SO reductase could play a role in the oxidative stress response of S. Typhimurium. MsrC is a methionine sulfoxide reductase previously shown to be specific for free Met-R-SO in Escherichia (E.) coli. We tested a ΔmsrC single mutant and a ΔmsrBΔmsrC double mutant under various stress conditions, and found that MsrC is essential for survival of S. Typhimurium following exposure to H2O2, as well as for growth in macrophages, and in mice. Hence, this study demonstrates that all three methionine sulfoxide reductases, MsrA, MsrB and MsrC, facilitate growth of a canonical intracellular pathogen during infection. Interestingly MsrC is specific for the repair of free methionine sulfoxide, pointing to an important role of this pathway in the oxidative stress response of Salmonella Typhimurium. PMID:22073230

  20. Kinetic characteristics of ZENECA ZD5522, a potent inhibitor of human and bovine lens aldose reductase.

    PubMed

    Cook, P N; Ward, W H; Petrash, J M; Mirrlees, D J; Sennitt, C M; Carey, F; Preston, J; Brittain, D R; Tuffin, D P; Howe, R

    1995-04-18

    Aldose reductase (aldehyde reductase 2) catalyses the conversion of glucose to sorbitol, and methylglyoxal to acetol. Treatment with aldose reductase inhibitors (ARIs) is a potential approach to decrease the development of diabetic complications. The sulphonylnitromethanes are a recently discovered class of aldose reductase inhibitors, first exemplified by ICI215918. We now describe enzyme kinetic characterization of a second sulphonylnitromethane, 3',5'-dimethyl-4'-nitromethylsulphonyl-2-(2-tolyl)acetanilide (ZD5522), which is at least 10-fold more potent against bovine lens aldose reductase in vitro and which also has a greater efficacy for reduction of rat nerve sorbitol levels in vivo (ED95 = 2.8 mg kg-1 for ZD5522 and 20 mg kg-1 for ICI 215918). ZD5522 follows pure noncompetitive kinetics against bovine lens aldose reductase when either glucose or methylglyoxal is varied (K(is) = K(ii) = 7.2 and 4.3 nM, respectively). This contrasts with ICI 215918 which is an uncompetitive inhibitor (K(ii) = 100 nM) of bovine lens aldose reductase when glucose is varied. Against human recombinant aldose reductase, ZD5522 displays mixed noncompetitive kinetics with respect to both substrates (K(is) = 41 nM, K(ii) = 8 nM with glucose and K(is) = 52 nM, K(ii) = 3.8 nM with methylglyoxal). This is the first report of the effects of a sulphonylnitromethane on either human aldose reductase or utilization of methylglyoxal. These results are discussed with reference to a Di Iso Ordered Bi Bi mechanism for aldose reductase, where the inhibitors compete with binding of both the aldehyde substrate and alcohol product. This model may explain why aldose reductase inhibitors follow noncompetitive or uncompetitive kinetics with respect to aldehyde substrates, and X-ray crystallography paradoxically locates an ARI within the substrate binding site. Aldehyde reductase (aldehyde reductase 1) is closely related to aldose reductase. Inhibition of bovine kidney aldehyde reductase by ZD5522

  1. Functional properties and structural characterization of rice δ1-pyrroline-5-carboxylate reductase

    SciTech Connect

    Forlani, Giuseppe; Bertazzini, Michele; Zarattini, Marco; Funck, Dietmar; Ruszkowski, Milosz; Nocek, Bogusław

    2015-07-28

    The majority of plant species accumulate high intracellular levels of proline to cope with hyperosmotic stress conditions. Proline synthesis from glutamate is tightly regulated at both the transcriptional and the translational levels, yet little is known about the mechanisms for post-translational regulation of the enzymatic activities involved. The gene coding in rice (Oryza sativa L.) for δ1-pyrroline-5-carboxylate (P5C) reductase, the enzyme that catalyzes the second and final step in this pathway, was isolated and expressed in Escherichia coli. The structural and functional properties of the affinity-purified protein were characterized. As for most species, rice P5C reductase was able to use in vitro either NADH or NADPH as the electron donor. However, strikingly different effects of cations and anions were found depending on the pyridine nucleotide used, namely inhibition of NADH-dependent activity and stimulation of NADPH-dependent activity. Moreover, physiological concentrations of proline and NADP+ were strongly inhibitory for the NADH-dependent reaction, whereas the NADPH-dependent activity was mildly affected. Our results suggest that only NADPH may be used in vivo and that stress-dependent variations in ion homeostasis and NADPH/NADP+ ratio could modulate enzyme activity, being functional in promoting proline accumulation and potentially also adjusting NADPH consumption during the defense against hyperosmotic stress. The apparent molecular weight of the native protein observed in size exclusion chromatography indicated a high oligomerization state. We also report the first crystal structure of a plant P5C reductase at 3.40-Å resolution, showing a decameric quaternary assembly. It was possible to identify dynamic structural differences among rice, human, and bacterial enzymes.

  2. Nitric Oxide (NO) Generation from Heme/Copper Assembly Mediated Nitrite Reductase Activity

    PubMed Central

    Hematian, Shabnam; Siegler, Maxime A.

    2014-01-01

    Nitric oxide (NO) as a cellular signaling molecule and vasodilator regulates a range of physiological and pathological processes. Nitrite (NO2−) is recycled in vivo to generate nitric oxide, particularly in physiologic hypoxia and ischemia. The cytochrome c oxidase (CcO) binuclear hemea3/CuB active site is one entity known to be responsible for cellular nitrite conversion to nitric oxide. We recently reported that a partially reduced heme/Cu assembly reduces nitrite ion, producing NO; the heme serves as the reductant and cupric ion provides a Lewis Acid interaction with nitrite, facilitating nitrite (N−O) bond cleavage (Hematian et al., J Am Chem Soc 134:18912–18915, 2012). To further investigate this nitrite reductase (NIR) chemistry, copper(II)-nitrito complexes with tri-and tetra-dentate ligands were used in this study, where either O,O'-bidentate or O-unidentate modes of nitrite binding to the cupric center are present. To study the role of the reducing ability of the ferrous heme center, two different tetraarylporphyrinate-iron(II) complexes, one with electron donating para-methoxy peripheral substituents, (TMPP)FeII, and the other with electron withdrawing 2,6-difluorophenyl substituents, (F8)FeII, were employed. The results show that differing nitrite coordination modes to copper(II) ion leads to varying kinetic behavior. Here, also, the ferrous heme is in all cases the source of the reducing equivalent required to take nitrite to nitric oxide, but the reduction ability of the heme center does not play a key role in the observed overall reaction rate. Based on our observations, reaction mechanisms are proposed and discussed in terms of heme/Cu heterobinuclear structures. PMID:24430198

  3. Nitric oxide generation from heme/copper assembly mediated nitrite reductase activity.

    PubMed

    Hematian, Shabnam; Siegler, Maxime A; Karlin, Kenneth D

    2014-06-01

    Nitric oxide (NO) as a cellular signaling molecule and vasodilator regulates a range of physiological and pathological processes. Nitrite (NO2 (-)) is recycled in vivo to generate nitric oxide, particularly in physiologic hypoxia and ischemia. The cytochrome c oxidase binuclear heme a 3/CuB active site is one entity known to be responsible for conversion of cellular nitrite to nitric oxide. We recently reported that a partially reduced heme/copper assembly reduces nitrite ion, producing nitric oxide; the heme serves as the reductant and the cupric ion provides a Lewis acid interaction with nitrite, facilitating nitrite (N-O) bond cleavage (Hematian et al., J. Am. Chem. Soc. 134:18912-18915, 2012). To further investigate this nitrite reductase chemistry, copper(II)-nitrito complexes with tridentate and tetradentate ligands were used in this study, where either O,O'-bidentate or O-unidentate modes of nitrite binding to the cupric center are present. To study the role of the reducing ability of the ferrous heme center, two different tetraarylporphyrinate-iron(II) complexes, one with electron-donating para-methoxy peripheral substituents and the other with electron-withdrawing 2,6-difluorophenyl substituents, were used. The results show that differing modes of nitrite coordination to the copper(II) ion lead to differing kinetic behavior. Here, also, the ferrous heme is in all cases the source of the reducing equivalent required to convert nitrite to nitric oxide, but the reduction ability of the heme center does not play a key role in the observed overall reaction rate. On the basis of our observations, reaction mechanisms are proposed and discussed in terms of heme/copper heterobinuclear structures.

  4. Realgar- and cinnabar-containing an-gong-niu-huang wan (AGNH) is much less acutely toxic than sodium arsenite and mercuric chloride.

    PubMed

    Lu, Yuan-Fu; Yan, Jun-Wen; Wu, Qin; Shi, Jing-Zhen; Liu, Jie; Shi, Jing-Shan

    2011-01-15

    An-gong-niu-huang wan (AGNH) is a famous traditional Chinese medicine used for brain trauma, hemorrhage, and coma. AGNH contains 10% realgar (As₄S₄) and 10% cinnabar (HgS). Both As and Hg are well-known for their toxic effects, and the safety of AGNH is of concern. To address this question, the acute toxicity of AGNH, realgar and cinnabar were compared to sodium arsenite (NaAsO₂) and mercuric chloride (HgCl₂). Mice were administrated orally AGNH at 1, 3 and 6g/kg. AGNH at 3g/kg contains 2.8mmol As/kg as realgar and 1.18mmol Hg/kg as cinnabar. Realgar, cinnabar, arsenite (0.28 mmol/kg, 10% of realgar) and HgCl₂ (0.256 mmol/kg, 20% of cinnabar) were orally given to mice for comparison. Blood and tissues were collected 8h later for toxicity evaluation. Serum alanine aminotransferase was increased by arsenite and blood urea nitrogen was increased by HgCl₂. Total As accumulation after arsenite in liver (100-fold) and kidney (13-fold) was much higher than that after realgar. The accumulation of Hg after HgCl₂ in liver was 400-fold higher and kidney 30-fold higher than after cinnabar. Histopathology showed moderate liver and kidney injuries after arsenite and HgCl₂, but injuries were mild or absent after AGNH, realgar, and cinnabar. The expression of metallothionein-1, a biomarker of metal exposure, was increased 4-10-fold by arsenite and HgCl₂, but was unchanged by AGNH, realgar and cinnabar. Thus, AGNH, realgar and cinnabar are much less toxic acutely than arsenite and HgCl₂. The chemical forms of As and Hg are extremely important factors in determining their disposition and toxicity. Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.

  5. Alteration of mercuric chloride-induced autoimmune glomerulonephritis in brown-Norway rats by herring oil, evening primrose oil and OKY-046 a selective TXA-synthetase inhibitor.

    PubMed

    Papanikolaou, N

    1987-05-01

    Repeated subcutaneous (SC) injections of mercuric chloride (MC) in Brown Norway (BN) rats induce an autoimmune glomerulonephritis (GN) due to antiglomerular basement membrane (BM) antibody deposition in the glomeruli. The aim of this study was to investigate the effects on MC-induced autoimmune GN of OKY-046, a selective TXA-synthetase inhibitor herring oil (HO), which is rich in eicosapentaenoic acid (EPA) (5.6%) precursor of the three series of prostaglandins (PGs) and of (inactive) thromboxane (TXA3), and evening primrose oil (EPO), which is rich in linoleic acid (LA) (72%) and gamma-linolenic acid (GLNA) (9%), precursors of the one series of PGs, mainly PGE1, and of (inactive) TXA1. The administration of OKY-046 significantly inhibited proteinuria, partially prevented fibrin thrombi (FT) formation in the glomeruli, decreased urinary TXB, enhanced 6ketoPGF excretion and, increased survival rate of the animals from 60% (group receiving only MC) to 86%. However, OKY-046 did not prevent body weight (BW) loss or the development and deposition of IgG in the glomeruli. Increased intake of HO (80 days prior and throughout the experiment) and avoidance of arachidonic acid (AA) intake produced an effect comparable to that of OKY-046 in the rats. Furthermore, HO significantly inhibited the deposition of IgG in the glomeruli, increased the survival rate of the animals to 100% and further enhanced the increased urinary PGE excretion induced by MC. However, HO did not prevent BW loss in the animals. Increased intake of EPO and avoidance of AA intake produced an effect comparable to that of HO. Additionally, EPO completely prevented BW loss induced by MC in these animals. These findings suggest that the metabolites of AA, EPA and GLNA play an important role either in the development or in the modulation of this model of MC induced GN.

  6. Attenuation by methyl mercury and mercuric sulfide of pentobarbital induced hypnotic tolerance in mice through inhibition of ATPase activities and nitric oxide production in cerebral cortex.

    PubMed

    Chuu, Jiunn-Jye; Huang, Zih-Ning; Yu, Hsun-Hsin; Chang, Liang-Hao; Lin-Shiau, Shoei-Yn

    2008-06-01

    This study is aimed at exploring the possible mechanism of hypnosis-enhancing effect of HgS or cinnabar (a traditional Chinese medicine containing more than 95% HgS) in mice treated with pentobarbital. We also examined whether the effect of HgS is different from that of the well-known methyl mercury (MeHg). After a short period (7 days) of oral administration to mice, a nontoxic dose (0.1 g/kg) of HgS not only significantly enhanced pentobarbital-induced hypnosis but also attenuated tolerance induction; while a higher dose (1 g/kg) of HgS or cinnabar exerted an almost irreversible enhancing effect on pentobarbital-hypnosis similar to that of MeHg (2 mg/kg) tested, which was still effective even after 10 or 35 days cessation of administration. To study comparatively the effects of different mercury forms from oral administration of MeHg and HgS on membrane ATPase activities of experimental mice, analysis of the Hg content in the cerebral cortex revealed that correlated with the decrease of Na(+)/K(+)-ATPase and Ca(2+)-ATPase activities. Furthermore, NO levels of blood but not that of cerebral cortex were also decreased by mercuric compounds. Although pentobarbital alone enhanced cytochrome p450-2C9 in time dependent manner, all of mercurial compounds tested had no such effect. All of these findings indicated that the mercurial compounds including cinnabar, HgS and MeHg exert a long-lasting enhancing hypnotic activity without affecting pentobarbital metabolism, which provides evidence-based sedative effect of cinnabar used in Chinese traditional medicine for more than 2,000 years. The nontoxic HgS dosing (0.1 g/kg/day) for consecutive 7 days is perhaps useful for delaying or preventing pentobarbital-tolerance.

  7. Physiological studies on the effect of copper nicotinate (Cu-N complex) on the fish, Clarias gariepinus, exposed to mercuric chloride.

    PubMed

    Al-Salahy, M Bassam

    2011-09-01

    Female catfish, Clarias gariepinus, were collected from the Nile River at Assiut region, were divided into 7 groups. The first group was left as control, and the second was treated with mercuric chloride (MC) for 3 weeks following by normal water for 1 week. The third, fourth and fifth groups were provided by MC (150 μg/ l of water). This treatment was continued for 3 weeks. Then, the fish were received CN instead of MC, for 1 week, with 15 and 25 mg CN/100 g wet food. The fifth fish group received diet supplemented with vit E (α-tocopherol) (100 mg/kg wet diet), for 1 week, instead of MC treatment. Vitamin E was used as standard antioxidant drug. Following 3 weeks of normal ambient water, the sixth and seventh aquaria received only CN for 1 week, with 15 and 25 mg CN respectively/100 g wet food, respectively. At the end of the experiment, Samples of liver, kidneys (posterior part), gills (right gills) and ovary were excised. The measurement included the oxidative stress parameters: carbonyl protein and total peroxide and the antioxidant enzyme activities superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST) in all selected organs. MC treatment induced harmful effect in fish, probably due to its enhancing effect on reactive oxygen species (ROS) production in fish organs especially the respiratory and osmoregulatory organs namely gills. The result suggests that this gill damage may exert hypoxic case, anoxia for different organs and some Cu excretion resulting in a magnification of ROS overproduction. Also, the observed oxidative stress in ovary tissue of MC-treated fish may affect fish fertility. The addition of CN in fish diets could protect the fish C. gariepinus against MC-induced oxidative damage showing recovery of fish organs. It could suggest that the detoxifying mechanism of action of CN is mainly due to its scavenging activity of free radicals rather than tissue healing.

  8. Toxicometabolomics approach to urinary biomarkers for mercuric chloride (HgCl{sub 2})-induced nephrotoxicity using proton nuclear magnetic resonance ({sup 1}H NMR) in rats

    SciTech Connect

    Kim, Kyu-Bong; Um, So Young; Chung, Myeon Woo; Jung, Seung Chul; Oh, Ji Seon; Kim, Seon Hwa; Na, Han Sung; Lee, Byung Mu; Choi, Ki Hwan

    2010-12-01

    The primary objective of this study was to determine and characterize surrogate biomarkers that can predict nephrotoxicity induced by mercuric chloride (HgCl{sub 2}) using urinary proton nuclear magnetic resonance ({sup 1}H NMR) spectral data. A procedure for {sup 1}H NMR urinalysis using pattern recognition was proposed to evaluate nephrotoxicity induced by HgCl{sub 2} in Sprague-Dawley rats. HgCl{sub 2} at 0.1 or 0.75 mg/kg was administered intraperitoneally (i.p.), and urine was collected every 24 h for 6 days. Animals (n = 6 per group) were sacrificed 3 or 6 days post-dosing in order to perform clinical blood chemistry tests and histopathologic examinations. Urinary {sup 1}H NMR spectroscopy revealed apparent differential clustering between the control and HgCl{sub 2} treatment groups as evidenced by principal component analysis (PCA) and partial least square (PLS)-discriminant analysis (DA). Time- and dose-dependent separation of HgCl{sub 2}-treated animals from controls was observed by PCA of {sup 1}H NMR spectral data. In HgCl{sub 2}-treated rats, the concentrations of endogenous urinary metabolites of glucose, acetate, alanine, lactate, succinate, and ethanol were significantly increased, whereas the concentrations of 2-oxoglutarate, allantoin, citrate, formate, taurine, and hippurate were significantly decreased. These endogenous metabolites were selected as putative biomarkers for HgCl{sub 2}-induced nephrotoxicity. A dose response was observed in concentrations of lactate, acetate, succinate, and ethanol, where severe disruption of the concentrations of 2-oxoglutarate, citrate, formate, glucose, and taurine was observed at the higher dose (0.75 mg/kg) of HgCl{sub 2}. Correlation of urinary {sup 1}H NMR PLS-DA data with renal histopathologic changes suggests that {sup 1}H NMR urinalysis can be used to predict or screen for HgCl{sub 2}-induced nephrotoxicity{sub .}

  9. The strain difference in the effect of mercuric chloride on antigen-triggered serotonin release from rat mast cells is not mediated via interferon-gamma.

    PubMed Central

    Hodson, D; Oliveira, D B

    1996-01-01

    Previous work has shown that in vitro exposure of Brown-Norway (BN) rat peritoneal mast cells to mercuric chloride (HgCl2) causes enhancement of subsequent mediator release induced by cross-linking of surface immunoglobulin E (IgE). This enhancing effect is seen significantly less often with peritoneal cells from Lewis rats. In addition HgCl2 has been shown to suppress interferon (IFN)-gamma production by BN but not Lewis splenocytes. Given that IFN-gamma is known to inhibit mediator release by mast cells, we hypothesized that the strain difference in the effect of HgCl2 on mediator release was mediated via a differential effect on IFN-gamma release from T cells in the mixed peritoneal cell population: IFN-gamma release would be suppressed in the case of the BN rat, releasing the mast cells from inhibition and resulting in the enhancing effect of HgCl2. The aim of the study was to test two predictions of this hypothesis. Exposure of BN rat mast cells to IFN-gamma inhibited subsequent antigen-induced mediator release but did not significantly reduce HgCl2-mediated enhancement of this release. Exposure of Lewis rat mast cells to blocking concentrations of anti-IFN-gamma did not reveal any HgCl2-mediated enhancement of mediator release. These observations provide strong evidence against the hypothesis that the differential effects of HgCl2 on BN and Lewis rat mast cells are mediated via IFN-gamma. In addition the results revealed that BN rat mast cells are significantly more sensitive than Lewis rat mast cells to the inhibitory effects of IFN-gamma on antigen-induced mediator release. PMID:8958063

  10. Inclusion of Ethyl Acetoacetate Bearing 7-Hydroxycoumarin Dye by β-Cyclodextrin and its Cooperative Assembly with Mercury(II) Ions: Spectroscopic and Molecular Modeling Studies.

    PubMed

    Aliaga, Margarita E; Fierro, Angélica; Uribe, Iván; García-Río, Luis; Cañete, Álvaro

    2016-10-18

    The inclusion of the fluorescent organic dye, ethyl 3-(7-hydroxy-2-oxo-2H-chromen-3-yl)-3-oxopropanoate (1) by the host β-cyclodextrin (β-CD), and its response toward mercuric ions (Hg(2+) ), was studied by UV/Vis, fluorescence, and (1) H NMR spectroscopic analyses, mass spectrometry and molecular modeling studies. (1) H NMR measurements together with molecular modeling studies for dye 1 demonstrate that it exhibits two tautomeric forms (keto and enol); however, when the dye is included into the β-CD cavity, the enol form predominates. Moreover, by using spectroscopic and spectrometry techniques, a 1:1 stoichiometry was determined for the complexes formed between dye 1 (enol form) and β-CD, with a binding constant (Kb1 =1.8×10(4)  m(-1) ) and for the dye 1 (keto form)-Hg(2+) (Kb2 =2.3×10(3)  m(-1) ). Interestingly, in the presence of 1-β-CD complex and mercuric ions, a ternary supramolecular system (Hg-1-β-CD complex) was established, with a 1:1:1 stoichiometry and a Kb3 value of 4.3×10(3)  m(-1) , with the keto form of the dye being the only one present in this assembly. The three-component system provides a starting point for the development of novel and directed supramolecular assemblies.

  11. Structure of the Molybdenum Site of EEcherichia Coli Trimethylamine N-Oxide Reductase

    SciTech Connect

    Zhang, L.; Nelson, K.Johnson; Rajagopalan, K.V.; George, G.N.

    2009-05-28

    We report a structural characterization of the molybdenum site of recombinant Escherichia coli trimethylamine N-oxide (TMAO) reductase using X-ray absorption spectroscopy. The enzyme active site shows considerable similarity to that of dimethyl sulfoxide (DMSO) reductase, in that, like DMSO reductase, the TMAO reductase active site can exist in multiple forms. Examination of the published crystal structure of TMAO oxidase from Shewanella massilia indicates that the postulated Mo coordination structure is chemically impossible. The presence of multiple active site structures provides a potential explanation for the anomalous features reported from the crystal structure.

  12. A flavone from Manilkara indica as a specific inhibitor against aldose reductase in vitro.

    PubMed

    Haraguchi, Hiroyuki; Hayashi, Ryosuke; Ishizu, Takashi; Yagi, Akira

    2003-09-01

    Isoaffinetin (5,7,3',4',5'-pentahydroxyflavone-6-C-glucoside) was isolated from Manilkara indica as a potent inhibitor of lens aldose reductase by bioassay-directed fractionation. This C-glucosyl flavone showed specific inhibition against aldose reductases (rat lens, porcine lens and recombinant human) with no inhibition against aldehyde reductase and NADH oxidase. Kinetic analysis showed that isoaffinetin exhibited uncompetitive inhibition against both dl-glyceraldehyde and NADPH. A structure-activity relationship study revealed that the increasing number of hydroxy groups in the B-ring contributes to the increase in aldose reductase inhibition by C-glucosyl flavones.

  13. Ammonification in Bacillus subtilis Utilizing Dissimilatory Nitrite Reductase Is Dependent on resDE

    PubMed Central

    Hoffmann, Tamara; Frankenberg, Nicole; Marino, Marco; Jahn, Dieter

    1998-01-01

    During anaerobic nitrate respiration Bacillus subtilis reduces nitrate via nitrite to ammonia. No denitrification products were observed. B. subtilis wild-type cells and a nitrate reductase mutant grew anaerobically with nitrite as an electron acceptor. Oxygen-sensitive dissimilatory nitrite reductase activity was demonstrated in cell extracts prepared from both strains with benzyl viologen as an electron donor and nitrite as an electron acceptor. The anaerobic expression of the discovered nitrite reductase activity was dependent on the regulatory system encoded by resDE. Mutation of the gene encoding the regulatory Fnr had no negative effect on dissimilatory nitrite reductase formation. PMID:9422613

  14. The inhibitory activity of aldose reductase in vitro by constituents of Garcinia mangostana Linn.

    PubMed

    Fatmawati, Sri; Ersam, Taslim; Shimizu, Kuniyoshi

    2015-01-15

    We investigated aldose reductase inhibition of Garcinia mangostana Linn. from Indonesia. Dichloromethane extract of the root bark of this tree was found to demonstrate an IC50 value of 11.98 µg/ml for human aldose reductase in vitro. From the dichloromethane fraction, prenylated xanthones were isolated as potent human aldose reductase inhibitors. We discovered 3-isomangostin to be most potent against aldose reductase, with an IC50 of 3.48 µM. Copyright © 2014 Elsevier GmbH. All rights reserved.

  15. Identification of the 7-Hydroxymethyl Chlorophyll a Reductase of the Chlorophyll Cycle in Arabidopsis[W

    PubMed Central

    Meguro, Miki; Ito, Hisashi; Takabayashi, Atsushi; Tanaka, Ryouichi; Tanaka, Ayumi

    2011-01-01

    The interconversion of chlorophyll a and chlorophyll b, referred to as the chlorophyll cycle, plays a crucial role in the processes of greening, acclimation to light intensity, and senescence. The chlorophyll cycle consists of three reactions: the conversions of chlorophyll a to chlorophyll b by chlorophyllide a oxygenase, chlorophyll b to 7-hydroxymethyl chlorophyll a by chlorophyll b reductase, and 7-hydroxymethyl chlorophyll a to chlorophyll a by 7-hydroxymethyl chlorophyll a reductase. We identified 7-hydroxymethyl chlorophyll a reductase, which is the last remaining unidentified enzyme of the chlorophyll cycle, from Arabidopsis thaliana by genetic and biochemical methods. Recombinant 7-hydroxymethyl chlorophyll a reductase converted 7-hydroxymethyl chlorophyll a to chlorophyll a using ferredoxin. Both sequence and biochemical analyses showed that 7-hydroxymethyl chlorophyll a reductase contains flavin adenine dinucleotide and an iron-sulfur center. In addition, a phylogenetic analysis elucidated the evolution of 7-hydroxymethyl chlorophyll a reductase from divinyl chlorophyllide vinyl reductase. A mutant lacking 7-hydroxymethyl chlorophyll a reductase was found to accumulate 7-hydroxymethyl chlorophyll a and pheophorbide a. Furthermore, this accumulation of pheophorbide a in the mutant was rescued by the inactivation of the chlorophyll b reductase gene. The downregulation of pheophorbide a oxygenase activity is discussed in relation to 7-hydroxymethyl chlorophyll a accumulation. PMID:21934147

  16. Components of glycine reductase from Eubacterium acidaminophilum. Cloning, sequencing and identification of the genes for thioredoxin reductase, thioredoxin and selenoprotein PA.

    PubMed

    Lübbers, M; Andreesen, J R

    1993-10-15

    The genes encoding thioredoxin reductase (trxB), thioredoxin (trxA), protein PA of glycine reductase (grdA) and the first 23 amino acids of the large subunit of protein PC of glycine reductase (grdC) belonging to the reductive deamination systems present in Eubacterium acidaminophilum were cloned and sequenced. The proteins were products of closely linked genes with 314 codons (thioredoxin reductase), 110 codons (thioredoxin), and 158 codons (protein PA). The protein previously called 'atypically small lipoamide dehydrogenase' or 'electron transferring flavoprotein' could now conclusively be identified as a thioredoxin reductase (subunit mass of 34781 Da) by the alignment with the enzyme of Escherichia coli showing the same typical order of the corresponding domains. The thioredoxin (molecular mass of 11742 Da) deviated considerably from the known consensus sequence, even in the most strongly conserved redox-active segment WCGPC that was now GCVPC. The selenocysteine of protein PA (molecular mass of 16609 Da) was encoded by TGA. The protein was highly similar to those of Clostridium purinolyticum and Clostridium sticklandii involved in glycine reductase. Thioredoxin reductase and thioredoxin of E. acidaminophilum could be successfully expressed in E. coli.

  17. Insights into function, catalytic mechanism, and fold evolution of selenoprotein methionine sulfoxide reductase B1 through structural analysis.

    PubMed

    Aachmann, Finn L; Sal, Lena S; Kim, Hwa-Young; Marino, Stefano M; Gladyshev, Vadim N; Dikiy, Alexander

    2010-10-22

    Methionine sulfoxide reductases protect cells by repairing oxidatively damaged methionine residues in proteins. Here, we report the first three-dimensional structure of the mammalian selenoprotein methionine sulfoxide reductase B1 (MsrB1), determined by high resolution NMR spectroscopy. Heteronuclear multidimensional spectra yielded NMR spectral assignments for the reduced form of MsrB1 in which catalytic selenocysteine (Sec) was replaced with cysteine (Cys). MsrB1 consists of a central structured core of two β-sheets and a highly flexible, disordered N-terminal region. Analysis of pH dependence of NMR signals of catalytically relevant residues, comparison with the data for bacterial MsrBs, and NMR-based structural analysis of methionine sulfoxide (substrate) and methionine sulfone (inhibitor) binding to MsrB1 at the atomic level reveal a mechanism involving catalytic Sec(95) and resolving Cys(4) residues in catalysis. The MsrB1 structure differs from the structures of Cys-containing MsrBs in the use of distal selenenylsulfide, residues needed for catalysis, and the mode in which the active form of the enzyme is regenerated. In addition, this is the first structure of a eukaryotic zinc-containing MsrB, which highlights the structural role of this metal ion bound to four conserved Cys. We integrated this information into a structural model of evolution of MsrB superfamily.

  18. Charaterization of bumarsin, a 3-hydroxy-3-methylglutaryl-coenzyme reductase inhibitor from Mesobuthus martensii Karsch venom.

    PubMed

    Chai, S C; Armugam, A; Strong, P N; Jeyaseelan, K

    2012-09-01

    Scorpion venoms are rich sources of bioactive peptides and are widely known for their ion channel inhibiting properties. We have isolated, cloned and characterized a venom protein (Bumarsin) from the Chinese scorpion, Mesobuthus martensii Karsch. Bumarsin cDNA encodes a 8132 Da, 72 amino acid mature protein that most probably exists in its native form as a Cys-bridged homodimer. We have identified this novel protein to be an inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase activity. 0.6 μM of Bumarsin inhibits 32% of the HMG-CoA reductase activity, in comparison to 10 μM simvastatin which only inhibits 35% of the activity. RT-PCR and SELDI-TOF mass spectrometric studies demonstrate that bumarsin regulates the expression of both genes and proteins involved in cholesterol homeostasis. Our results suggest that bumarsin may provide a model for the design of novel drugs that can be used to modulate cholesterol homeostasis. Copyright © 2012 Elsevier Ltd. All rights reserved.

  19. Ultrasound-assisted dispersive magnetic solid phase extraction for preconcentration and determination of trace amount of Hg (II) ions from food samples and aqueous solution by magnetic graphene oxide (Fe3O4@GO/2-PTSC): Central composite design optimization.

    PubMed

    Keramat, Akram; Zare-Dorabei, Rouholah

    2017-09-01

    In this work, the synthesis of the magnetic graphene oxide modified by 2-pyridinecarboxaldehyde thiosemicarbazone groups (Fe3O4@GO/2-PTSC) was utilized for preconcentration and determination of mercuric ions in a trace amount by inductively coupled plasma-optical emission spectrometry (ICP-OES). Characterization of the adsorbent was performed using various techniques, such as FT-IR, VSM, SEM and XRD analysis. Central composite design (CCD) under response surface methodology (RSM) was used for obtaining the most important parameters and probable interactions in variables. The variables such as adsorbent dosage, pH, desorption time, and eluent volume was optimized. These values were 8mg, 5.4min, 0.5mL (HCl, 0.1M), respectively. Sonication had an important role in shortening the adsorption time of Hg (II) ions by enhancing the dispersion of adsorbent in solution. Under the optimal conditions, the proposed method presented high enrichment factor of 193, an extraction percentage of 96.5, a detection limit of 0.0079µgL(-1) and a relative standard deviation (RSD %) of 1.63%. Finally, the application of the synthesized material was evaluated for preconcentration and determination of mercuric ions from foods and environmental waters samples. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Structural and Biochemical Characterization of Cinnamoyl-CoA Reductases.

    PubMed

    Sattler, Steven A; Walker, Alexander M; Vermerris, Wilfred; Sattler, Scott E; Kang, ChulHee

    2017-02-01

    Cinnamoyl-coenzyme A reductase (CCR) catalyzes the reduction of hydroxycinnamoyl-coenzyme A (CoA) esters using NADPH to produce hydroxycinnamyl aldehyde precursors in lignin synthesis. The catalytic mechanism and substrate specificity of cinnamoyl-CoA reductases from sorghum (Sorghum bicolor), a strategic plant for bioenergy production, were deduced from crystal structures, site-directed mutagenesis, and kinetic and thermodynamic analyses. Although SbCCR1 displayed higher affinity for caffeoyl-CoA or p-coumaroyl-CoA than for feruloyl-CoA, the enzyme showed significantly higher activity for the latter substrate. Through molecular docking and comparisons between the crystal structures of the Vitis vinifera dihydroflavonol reductase and SbCCR1, residues threonine-154 and tyrosine-310 were pinpointed as being involved in binding CoA-conjugated phenylpropanoids. Threonine-154 of SbCCR1 and other CCRs likely confers strong substrate specificity for feruloyl-CoA over other cinnamoyl-CoA thioesters, and the T154Y mutation in SbCCR1 led to broader substrate specificity and faster turnover. Through data mining using our structural and biochemical information, four additional putative CCR genes were discovered from sorghum genomic data. One of these, SbCCR2, displayed greater activity toward p-coumaroyl-CoA than did SbCCR1, which could imply a role in the synthesis of defense-related lignin. Taken together, these findings provide knowledge about critical residues and substrate preference among CCRs and provide, to our knowledge, the first three-dimensional structure information for a CCR from a monocot species.