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Sample records for methylococcus capsulatus bath

  1. Cytochrome c' of Methylococcus capsulatus Bath.

    PubMed

    Zahn, J A; Arciero, D M; Hooper, A B; Dispirito, A A

    1996-09-15

    Cytochrome c' was isolated from the obligate methylotroph Methylococcus capsulatus Bath. The native and subunit molecular masses of the cytochrome were 34.9 kDa and 16.2 kDa, respectively, with an isoelectric pH of 7.0. The amino acid composition and N-terminal amino acid sequence were consistent with identification of the protein as a cytochrome c'. The electron paramagnetic resonance spectrum of the monoheme cytochrome indicated the presence of a high spin, S = 5/2, heme center that is diagnostic of cytochromes c'. The optical absorption spectra of ferric or ferrous cytochrome c' were also characteristic of cytochromes c'. The ferrocytochrome bound carbon monoxide and nitric oxide, but not isocyanide, cyanide, or azide. Changes in physical properties due to binding of CO or NO to some other c'-type cytochromes have been interpreted as an indication of dimer dissociation. In the case of cytochrome c' from M. capsulatus Bath, analytical ultracentrifugation of the ferricytochrome, the ferrocytochrome, and the ferrocytochrome-CO complex indicate that the changes induced by binding of CO are conformational and are not consistent with dimer dissociation. EPR spectra show that cytochrome c' was reduced in the presence of hydroxylamine only when in a complex with cytochrome P-460. The value of the midpoint potential, Em 7.0, was -250 mV for cytochrome c' from M. capsulatus Bath, which is well below the range of values reported for other cytochromes c'. The values of midpoint potentials for cytochrome P-460 (Em 7.0 = -300 mV to -380 mV) and cytochrome C555 (Em 7.0 = +175 mV to +195 mV) are less than and greater than, respectively, the value for cytochrome c' and suggest the possibility that the latter may function as an electron shuttle between cytochrome P-460 and cytochrome C555. PMID:8856071

  2. Cytochrome c peroxidase from Methylococcus capsulatus Bath.

    PubMed

    Zahn, J A; Arciero, D M; Hooper, A B; Coats, J R; DiSpirito, A A

    1997-11-01

    A bacterial cytochrome c peroxidase was purified from the obligate methanotroph Methylococcus capsulatus Bath in either the fully oxidized or the half reduced form depending on the purification procedure. The cytochrome was a homo-dimer with a subunit mol mass of 35.8 kDa and an isoelectric point of 4.5. At physiological temperatures, the enzyme contained one high-spin, low-potential (Em7 = -254 mV) and one low-spin, high-potential (Em7 = +432 mM ) heme. The low-potential heme center exhibited a spin-state transition from the penta-coordinated, high-spin configuration to a low-spin configuration upon cooling the enzyme to cryogenic temperatures. Using M. capsulatus Bath ferrocytochrome c555 as the electron donor, the KM and Vmax for peroxide reduction were 510 +/- 100 nM and 425 +/- 22 mol ferrocytochrome c555 oxidized min-1 (mole cytochrome c peroxidase)-1, respectively. PMID:9325424

  3. Outer membrane proteins of Methylococcus capsulatus (Bath).

    PubMed

    Fjellbirkeland, A; Kleivdal, H; Joergensen, C; Thestrup, H; Jensen, H B

    1997-08-01

    Membranes obtained from whole-cell lysates of Methylococcus capsulatus (Bath) were separated by Triton X-100 extraction. The resulting insoluble fraction was enriched in outer membranes as assessed by electron microscopy and by the content of beta-hydroxy palmitic acid and particulate methane monooxygenase. Major proteins with molecular masses of approximately 27, 40, 46, 59, and 66 kDa were detected by SDS-PAGE of the Triton-X-100-insoluble membranes. MopA, MopB, MopC, MopD, and MopE (Methylococcus outer membrane protein) are proposed to designate these proteins. Several of the Mop proteins exhibited heat-modifiable properties in SDS-PAGE and were influenced by the presence of 2-mercaptoethanol in the sample buffer. The 46- and 59-kDa bands migrated as a single high-molecular-mass 95-kDa oligomer under mild denaturing conditions. When reconstituted into black lipid membranes, this oligomer was shown to serve as a channel with an estimated single-channel conductance of 1.4 nS in 1 M KCl. PMID:9238104

  4. Isolation, purification and characterization of hemerythrin from Methylococcus capsulatus (Bath).

    PubMed

    Kao, Wei-Chun; Wang, Vincent C-C; Huang, Yi-Che; Yu, Steve S-F; Chang, Ta-Chau; Chan, Sunney I

    2008-08-01

    Earlier work from our laboratory has indicated that a hemerythrin-like protein was over-produced together with the particulate methane monooxygenase (pMMO) when Methylococcus capsulatus (Bath) was grown under high copper concentrations. A homologue of hemerythrin had not previously been found in any prokaryote. To confirm its identity as a hemerythrin, we have isolated and purified this protein by ion-exchange, gel-filtration and hydrophobic interaction chromatography, and characterized it by mass spectrometry, UV-visible, CD, EPR and resonance Raman spectroscopy. On the basis of biophysical and multiple sequence alignment analysis, the protein isolated from M. capsulatus (Bath) is in accord with hemerythrins previously reported from higher organisms. Determination of the Fe content in conjunction with molecular-weight estimation and mass analysis indicates that the native hemerythrin in M. capsulatus (Bath) is a monomer with molecular mass 14.8 kDa, in contrast to hemerythrins from other eukaryotic organisms, where they typically exist as a tetramer or higher oligomers. PMID:18397812

  5. Cytochrome P460 genes from the methanotroph Methylococcus capsulatus bath.

    PubMed

    Bergmann, D J; Zahn, J A; Hooper, A B; DiSpirito, A A

    1998-12-01

    P460 cytochromes catalyze the oxidation of hydroxylamine to nitrite. They have been isolated from the ammonia-oxidizing bacterium Nitrosomonas europaea (R. H. Erickson and A. B. Hooper, Biochim. Biophys. Acta 275:231-244, 1972) and the methane-oxidizing bacterium Methylococcus capsulatus Bath (J. A. Zahn et al., J. Bacteriol. 176:5879-5887, 1994). A degenerate oligonucleotide probe was synthesized based on the N-terminal amino acid sequence of cytochrome P460 and used to identify a DNA fragment from M. capsulatus Bath that contains cyp, the gene encoding cytochrome P460. cyp is part of a gene cluster that contains three open reading frames (ORFs), the first predicted to encode a 59,000-Da membrane-bound polypeptide, the second predicted to encode a 12, 000-Da periplasmic protein, and the third (cyp) encoding cytochrome P460. The products of the first two ORFs have no apparent similarity to any proteins in the GenBank database. The overall sequence similarity of the P460 cytochromes from M. capsulatus Bath and N. europaea was low (24.3% of residues identical), although short regions of conserved residues are present in the two proteins. Both cytochromes have a C-terminal, c-heme binding motif (CXXCH) and a conserved lysine residue (K61) that may provide an additional covalent cross-link to the heme (D. M. Arciero and A. B. Hooper, FEBS Lett. 410:457-460, 1997). Gene probing using cyp indicated that a cytochrome P460 similar to that from M. capsulatus Bath may be present in the type II methanotrophs Methylosinus trichosporium OB3b and Methylocystis parvus OBBP but not in the type I methanotrophs Methylobacter marinus A45, Methylomicrobium albus BG8, and Methylomonas sp. strains MN and MM2. Immunoblot analysis with antibodies against cytochrome P460 from M. capsulatus Bath indicated that the expression level of cytochrome P460 was not affected either by expression of the two different methane monooxygenases or by addition of ammonia to the culture medium. PMID:9851984

  6. Response to mercury (II) ions in Methylococcus capsulatus (Bath).

    PubMed

    Boden, Rich; Murrell, J Colin

    2011-11-01

    The mercury (II) ion is toxic and is usually detoxified in Bacteria by reduction to elemental mercury, which is less toxic. This is catalysed by an NAD(P)H-dependent mercuric reductase (EC 1.16.1.1). Here, we present strong evidence that Methylococcus capsulatus (Bath) - a methanotrophic member of the Gammaproteobacteria - uses this enzyme to detoxify mercury. In radiorespirometry studies, it was found that cells exposed to mercury dissimilated 100% of [(14) C]-methane provided to generate reducing equivalents to fuel mercury (II) reduction, rather than the mix of assimilation and dissimilation found in control incubations. The detoxification system is constitutively expressed with a specific activity of 352 (±18) nmol NADH oxidized min(-1) (mg protein)(-1) . Putative mercuric reductase genes were predicted in the M. capsulatus (Bath) genome and found in mRNA microarray studies. The MerA-derived polypeptide showed high identity (> 80%) with MerA sequences from the Betaproteobacteria. PMID:22092810

  7. Methylococcus capsulatus (Bath) from genome to protein function, and vice versa.

    PubMed

    Karlsen, Odd A; Berven, Frode S; Bagstevold, June I; Larsen, Oivind; Jensen, Harald B

    2011-01-01

    The genome sequence of Methylococcus capsulatus (Bath), considered a model methylotroph, was published in 2004 [Ward, N., et al. (2004). Genomic insights into methanotrophy: the complete genome sequence of Methylococcus capsulatus (Bath). PLoS Biol.2, e303]. In the postgenomic era, the challenge is to determine the gene function, and to this end, genomics must be complemented with proteomic approaches. This chapter describes some experimental and computational approaches we have used and developed for the exploration of the genome and proteome of M. capsulatus (Bath). PMID:21419915

  8. Membrane-Associated Quinoprotein Formaldehyde Dehydrogenase from Methylococcus capsulatus Bath

    PubMed Central

    Zahn, James A.; Bergmann, David J.; Boyd, Jeffery M.; Kunz, Ryan C.; DiSpirito, Alan A.

    2001-01-01

    A membrane-associated, dye-linked formaldehyde dehydrogenase (DL-FalDH) was isolated from the obligate methylotroph Methylococcus capsulatus Bath. The enzyme was the major formaldehyde-oxidizing enzyme in cells cultured in high (above 1 μmol of Cu per mg of cell protein) copper medium and expressing the membrane-associated methane monooxygenase. Soluble NAD(P)+-linked formaldehyde oxidation was the major activity in cells cultured in low-copper medium and expressing the soluble methane monooxygenase (Tate and Dalton, Microbiology 145:159–167, 1999; Vorholt et al., J. Bacteriol. 180:5351–5356, 1998). The membrane-associated enzyme is a homotetramer with a subunit molecular mass of 49,500 Da. UV-visible absorption, electron paramagnetic resonance, and electrospray mass spectrometry suggest the redox cofactor of the DL-FalDH is pyrroloquinoline quinone (PQQ), with a PQQ-to-subunit stochiometry of approximately 1:1. The enzyme was specific for formaldehyde, oxidizing formaldehyde to formate, and utilized the cytochrome b559/569 complex as the physiological electron acceptor. PMID:11698372

  9. Membrane-associated quinoprotein formaldehyde dehydrogenase from Methylococcus capsulatus Bath.

    PubMed

    Zahn, J A; Bergmann, D J; Boyd, J M; Kunz, R C; DiSpirito, A A

    2001-12-01

    A membrane-associated, dye-linked formaldehyde dehydrogenase (DL-FalDH) was isolated from the obligate methylotroph Methylococcus capsulatus Bath. The enzyme was the major formaldehyde-oxidizing enzyme in cells cultured in high (above 1 micromol of Cu per mg of cell protein) copper medium and expressing the membrane-associated methane monooxygenase. Soluble NAD(P)(+)-linked formaldehyde oxidation was the major activity in cells cultured in low-copper medium and expressing the soluble methane monooxygenase (Tate and Dalton, Microbiology 145:159-167, 1999; Vorholt et al., J. Bacteriol. 180:5351-5356, 1998). The membrane-associated enzyme is a homotetramer with a subunit molecular mass of 49,500 Da. UV-visible absorption, electron paramagnetic resonance, and electrospray mass spectrometry suggest the redox cofactor of the DL-FalDH is pyrroloquinoline quinone (PQQ), with a PQQ-to-subunit stochiometry of approximately 1:1. The enzyme was specific for formaldehyde, oxidizing formaldehyde to formate, and utilized the cytochrome b(559/569) complex as the physiological electron acceptor. PMID:11698372

  10. Membrane-associated methane monooxygenase from Methylococcus capsulatus (Bath).

    PubMed Central

    Zahn, J A; DiSpirito, A A

    1996-01-01

    An active preparation of the membrane-associated methane monooxygenase (pMMO) from Methylococcus capsulatus Bath was isolated by ion-exchange and hydrophobic interaction chromatography using dodecyl beta-D-maltoside as the detergent. The active preparation consisted of three major polypeptides with molecular masses of 47,000, 27,000, and 25,000 Da. Two of the three polypeptides (those with molecular masses of 47,000 and 27,000 Da) were identified as the polypeptides induced when cells expressing the soluble MMO are switched to culture medium in which the pMMO is expressed. The 27,000-Da polypeptide was identified as the acetylene-binding protein. The active enzyme complex contained 2.5 iron atoms and 14.5 copper atoms per 99,000 Da. The electron paramagnetic resonance spectrum of the enzyme showed evidence for a type 2 copper center (g perpendicular = 2.057, g parallel = 2.24, and magnitude of A parallel = 172 G), a weak high-spin iron signal (g = 6.0), and a broad low-field (g = 12.5) signal. Treatment of the pMMO with nitric oxide produced the ferrous-nitric oxide derivative observed in the membrane fraction of cells expressing the pMMO. When duroquinol was used as a reductant, the specific activity of the purified enzyme was 11.1 nmol of propylene oxidized.min-1.mg of protein-1, which accounted for approximately 30% of the cell-free propylene oxidation activity. The activity was stimulated by ferric and cupric metal ions in addition to the cytochrome b-specific inhibitors myxothiazol and 2-heptyl-4-hydroxyquinoline-N-oxide. PMID:8576034

  11. Membrane-associated methane monooxygenase from Methylococcus capsulatus (Bath).

    PubMed

    Zahn, J A; DiSpirito, A A

    1996-02-01

    An active preparation of the membrane-associated methane monooxygenase (pMMO) from Methylococcus capsulatus Bath was isolated by ion-exchange and hydrophobic interaction chromatography using dodecyl beta-D-maltoside as the detergent. The active preparation consisted of three major polypeptides with molecular masses of 47,000, 27,000, and 25,000 Da. Two of the three polypeptides (those with molecular masses of 47,000 and 27,000 Da) were identified as the polypeptides induced when cells expressing the soluble MMO are switched to culture medium in which the pMMO is expressed. The 27,000-Da polypeptide was identified as the acetylene-binding protein. The active enzyme complex contained 2.5 iron atoms and 14.5 copper atoms per 99,000 Da. The electron paramagnetic resonance spectrum of the enzyme showed evidence for a type 2 copper center (g perpendicular = 2.057, g parallel = 2.24, and magnitude of A parallel = 172 G), a weak high-spin iron signal (g = 6.0), and a broad low-field (g = 12.5) signal. Treatment of the pMMO with nitric oxide produced the ferrous-nitric oxide derivative observed in the membrane fraction of cells expressing the pMMO. When duroquinol was used as a reductant, the specific activity of the purified enzyme was 11.1 nmol of propylene oxidized.min-1.mg of protein-1, which accounted for approximately 30% of the cell-free propylene oxidation activity. The activity was stimulated by ferric and cupric metal ions in addition to the cytochrome b-specific inhibitors myxothiazol and 2-heptyl-4-hydroxyquinoline-N-oxide. PMID:8576034

  12. Expression of Individual Copies of Methylococcus capsulatus Bath Particulate Methane Monooxygenase Genes

    PubMed Central

    Stolyar, Sergei; Franke, Marion; Lidstrom, Mary E.

    2001-01-01

    The expression of the two gene clusters encoding the particulate methane monooxygenase (pMMO) in Methylococcus capsulatus Bath was assessed by analysis of transcripts and by use of chromosomal gene fusions. The results suggest that the two clusters are functionally redundant but that relative expression alters depending on the copper levels available for growth. PMID:11160118

  13. Some properties of a soluble methane mono-oxygenase from Methylococcus capsulatus strain Bath.

    PubMed Central

    Colby, J; Dalton, H

    1976-01-01

    Soluble extracts of Methylococcus capsulatus (Bath), obtained by centrifugation of crude extracts at 160000g for 1h, catalyse the NAD(P)H- and O2-dependent disappearance of bromomethane, and also the formation of methanol from methane. Soluble methane mono-oxygenase is not inhibited by chelating agents or by most electron-transport inhibitors, and is a multicomponent enzyme. PMID:962879

  14. Expression of individual copies of Methylococcus capsulatus bath particulate methane monooxygenase genes.

    PubMed

    Stolyar, S; Franke, M; Lidstrom, M E

    2001-03-01

    The expression of the two gene clusters encoding the particulate methane monooxygenase (pMMO) in Methylococcus capsulatus Bath was assessed by analysis of transcripts and by use of chromosomal gene fusions. The results suggest that the two clusters are functionally redundant but that relative expression alters depending on the copper levels available for growth. PMID:11160118

  15. Computational and Experimental Analysis of the Secretome of Methylococcus capsulatus (Bath)

    PubMed Central

    Indrelid, Stine; Mathiesen, Geir; Jacobsen, Morten; Lea, Tor; Kleiveland, Charlotte R.

    2014-01-01

    The Gram-negative methanotroph Methylococcus capsulatus (Bath) was recently demonstrated to abrogate inflammation in a murine model of inflammatory bowel disease, suggesting interactions with cells involved in maintaining mucosal homeostasis and emphasizing the importance of understanding the many properties of M. capsulatus. Secreted proteins determine how bacteria may interact with their environment, and a comprehensive knowledge of such proteins is therefore vital to understand bacterial physiology and behavior. The aim of this study was to systematically analyze protein secretion in M. capsulatus (Bath) by identifying the secretion systems present and the respective secreted substrates. Computational analysis revealed that in addition to previously recognized type II secretion systems and a type VII secretion system, a type Vb (two-partner) secretion system and putative type I secretion systems are present in M. capsulatus (Bath). In silico analysis suggests that the diverse secretion systems in M.capsulatus transport proteins likely to be involved in adhesion, colonization, nutrient acquisition and homeostasis maintenance. Results of the computational analysis was verified and extended by an experimental approach showing that in addition an uncharacterized protein and putative moonlighting proteins are released to the medium during exponential growth of M. capsulatus (Bath). PMID:25479164

  16. Computational and experimental analysis of the secretome of Methylococcus capsulatus (Bath).

    PubMed

    Indrelid, Stine; Mathiesen, Geir; Jacobsen, Morten; Lea, Tor; Kleiveland, Charlotte R

    2014-01-01

    The Gram-negative methanotroph Methylococcus capsulatus (Bath) was recently demonstrated to abrogate inflammation in a murine model of inflammatory bowel disease, suggesting interactions with cells involved in maintaining mucosal homeostasis and emphasizing the importance of understanding the many properties of M. capsulatus. Secreted proteins determine how bacteria may interact with their environment, and a comprehensive knowledge of such proteins is therefore vital to understand bacterial physiology and behavior. The aim of this study was to systematically analyze protein secretion in M. capsulatus (Bath) by identifying the secretion systems present and the respective secreted substrates. Computational analysis revealed that in addition to previously recognized type II secretion systems and a type VII secretion system, a type Vb (two-partner) secretion system and putative type I secretion systems are present in M. capsulatus (Bath). In silico analysis suggests that the diverse secretion systems in M.capsulatus transport proteins likely to be involved in adhesion, colonization, nutrient acquisition and homeostasis maintenance. Results of the computational analysis was verified and extended by an experimental approach showing that in addition an uncharacterized protein and putative moonlighting proteins are released to the medium during exponential growth of M. capsulatus (Bath). PMID:25479164

  17. Molecular characterization of structural genes coding for a membrane bound hydrogenase in Methylococcus capsulatus (Bath).

    PubMed

    Csáki, R; Hanczár, T; Bodrossy, L; Murrell, J C; Kovács, K L

    2001-12-18

    The first gene cluster encoding for a membrane bound [NiFe] hydrogenase from a methanotroph, Methylococcus capsulatus (Bath), was cloned and sequenced. The cluster consisted of the structural genes hupS and hupL and accessory genes hupE, hupC and hupD. A DeltahupSL deletion mutant of Mc. capsulatus was constructed by marker exchange mutagenesis. Membrane associated hydrogenase activity disappeared. The membrane associated hydrogenase appeared to have a hydrogen uptake function in vivo. PMID:11750803

  18. The ribulose-1,5-bisphosphate carboxylase/oxygenase gene cluster of Methylococcus capsulatus (Bath).

    PubMed

    Baxter, Nardia J; Hirt, Robert P; Bodrossy, Levente; Kovacs, Kornel L; Embley, T Martin; Prosser, James I; Murrell, J Colin

    2002-04-01

    The genes encoding the ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) from Methylococcus capsulatus (Bath) were localised to an 8.3-kb EcoRI fragment of the genome. Genes encoding the large subunit ( cbbL), small subunit ( cbbS) and putative regulatory gene ( cbbQ) were shown to be located on one cluster. Surprisingly, cbbO, a second putative regulatory gene, was not located in the remaining 1.2-kb downstream (3') of cbbQ. However, probing of the M. capsulatus (Bath) genome with cbbO from Nitrosomonas europaea demonstrated that a cbbO homologue was contained within a separate 3.0-kb EcoRI fragment. Instead of a cbbR ORF being located upstream (5') of cbbL, there was a moxR-like ORF that was transcribed in the opposite direction to cbbL. There were three additional ORFs within the large 8.3-kb EcoRI fragment: a pyrE-like ORF, an rnr-like ORF and an incomplete ORF with no sequence similarity to any known protein. Phylogenetic analysis of cbbL from M. capsulatus (Bath) placed it within clade A of the green-type Form 1 Rubisco. cbbL was expressed in M. capsulatus (Bath) when grown with methane as a sole carbon and energy source under both copper-replete and copper-limited conditions. M. capsulatus (Bath) was capable of autotrophic growth on solid medium but not in liquid medium. Preliminarily investigations suggested that other methanotrophs may also be capable of autotrophic growth. Rubisco genes were also identified, by PCR, in Methylococcus-like strains and Methylocaldum species; however, no Rubisco genes were found in Methylomicrobium album BG8, Methylomonas methanica S1, Methylomonas rubra, Methylosinus trichosporium OB3b or Methylocystis parvus OBBP. PMID:11889481

  19. Effects of the non-commensal Methylococcus capsulatus Bath on mammalian immune cells.

    PubMed

    Christoffersen, Trine Eker; Olsen Hult, Lene Therese; Solberg, Henriette; Bakke, Anne; Kuczkowska, Katarzyna; Huseby, Eirin; Jacobsen, Morten; Lea, Tor; Kleiveland, Charlotte Ramstad

    2015-08-01

    Dietary inclusions of a bacterial meal consisting mainly of the non-commensal, methanotrophic bacteria Methylococcus capsulatus Bath have been shown to ameliorate symptoms of intestinal inflammation in different animal models. In order to investigate the molecular mechanisms causing these effects, we have studied the influence of this strain on different immune cells central for the regulation of inflammatory responses. Effects were compared to those induced by the closely related strain M. capsulatus Texas and the well-described probiotic strain Escherichia coli Nissle 1917. M. capsulatus Bath induced macrophage polarization toward a pro-inflammatory phenotype, but not to the extent observed after exposure to E. coli Nissle 1917. Likewise, dose-dependent abilities to activate NF-κB transcription in U937 cells were observed, with E. coli Nissle 1917 being most potent. High levels of CD141 on human primary monocyte-derived dendritic cells (moDCs) were only detected after exposure to E. coli Nissle 1917, which collectively indicate a superior capacity to induce Th1 cell responses for this strain. On the other hand, the M. capsulatus strains were more potent in increasing the expression of the maturation markers CD80, CD83 and CD86 than E. coli Nissle 1917. M. capsulatus Bath induced the highest levels of IL-6, IL-10 and IL-12 secretion from dendritic cells, suggesting that this strain generally the post potent inducer of cytokine secretion. These results show that M. capsulatus Bath exhibit immunogenic properties in mammalian in vitro systems which diverge from that of E. coli Nissle 1917. This may provide clues to how M. capsulatus Bath influence the adaptive immune system in vivo. However, further in vivo experiments are required for a complete understanding of how this strain ameliorates intestinal inflammation in animal models. PMID:25771177

  20. Structure and Protein–Protein Interactions of Methanol Dehydrogenase from Methylococcus capsulatus (Bath)

    PubMed Central

    2015-01-01

    In the initial steps of their metabolic pathway, methanotrophic bacteria oxidize methane to methanol with methane monooxygenases (MMOs) and methanol to formaldehyde with methanol dehydrogenases (MDHs). Several lines of evidence suggest that the membrane-bound or particulate MMO (pMMO) and MDH interact to form a metabolic supercomplex. To further investigate the possible existence of such a supercomplex, native MDH from Methylococcus capsulatus (Bath) has been purified and characterized by size exclusion chromatography with multi-angle light scattering and X-ray crystallography. M. capsulatus (Bath) MDH is primarily a dimer in solution, although an oligomeric species with a molecular mass of ∼450–560 kDa forms at higher protein concentrations. The 2.57 Å resolution crystal structure reveals an overall fold and α2β2 dimeric architecture similar to those of other MDH structures. In addition, biolayer interferometry studies demonstrate specific protein–protein interactions between MDH and M. capsulatus (Bath) pMMO as well as between MDH and the truncated recombinant periplasmic domains of M. capsulatus (Bath) pMMO (spmoB). These interactions exhibit KD values of 833 ± 409 nM and 9.0 ± 7.7 μM, respectively. The biochemical data combined with analysis of the crystal lattice interactions observed in the MDH structure suggest a model in which MDH and pMMO associate not as a discrete, stoichiometric complex but as a larger assembly scaffolded by the intracytoplasmic membranes. PMID:25185034

  1. Structure and protein-protein interactions of methanol dehydrogenase from Methylococcus capsulatus (Bath).

    PubMed

    Culpepper, Megen A; Rosenzweig, Amy C

    2014-10-01

    In the initial steps of their metabolic pathway, methanotrophic bacteria oxidize methane to methanol with methane monooxygenases (MMOs) and methanol to formaldehyde with methanol dehydrogenases (MDHs). Several lines of evidence suggest that the membrane-bound or particulate MMO (pMMO) and MDH interact to form a metabolic supercomplex. To further investigate the possible existence of such a supercomplex, native MDH from Methylococcus capsulatus (Bath) has been purified and characterized by size exclusion chromatography with multi-angle light scattering and X-ray crystallography. M. capsulatus (Bath) MDH is primarily a dimer in solution, although an oligomeric species with a molecular mass of ∼450-560 kDa forms at higher protein concentrations. The 2.57 Å resolution crystal structure reveals an overall fold and α2β2 dimeric architecture similar to those of other MDH structures. In addition, biolayer interferometry studies demonstrate specific protein-protein interactions between MDH and M. capsulatus (Bath) pMMO as well as between MDH and the truncated recombinant periplasmic domains of M. capsulatus (Bath) pMMO (spmoB). These interactions exhibit KD values of 833 ± 409 nM and 9.0 ± 7.7 μM, respectively. The biochemical data combined with analysis of the crystal lattice interactions observed in the MDH structure suggest a model in which MDH and pMMO associate not as a discrete, stoichiometric complex but as a larger assembly scaffolded by the intracytoplasmic membranes. PMID:25185034

  2. Direct electrochemistry of the hydroxylase of soluble methane monooxygenase from Methylococcus capsulatus (Bath).

    PubMed

    Kazlauskaite, J; Hill, H A; Wilkins, P C; Dalton, H

    1996-10-15

    The redox properties of the hydroxylase component of soluble methane monooxygenase from Methylococcus capsulatus (Bath) have been thoroughly investigated. Previous studies used redox indicator titrations and spectroscopic methods for the determination of the concentrations of reduced species. Herein we report, for the first time, direct electrochemistry (i.e. without the use of mediators) of the diiron centers of the hydroxylase from M. capsulatus (Bath) at a modified gold electrode giving rise to two waves at 4(+/- 10) mV and -386(+/- 14) mV versus saturated calomel electrode (SCE). In addition, the effects of proteins B and B' on the redox reactions were determined. The redox potentials of the complex with protein B are -25(+/- 14) mV and -433(+/- 8) mV versus SCE whereas protein B' had no effect though it did alter the effect of protein B on the redox potentials. PMID:8917455

  3. Nitrogen isotopomer site preference of N2O produced by Nitrosomonas europaea and Methylococcus capsulatus Bath.

    PubMed

    Sutka, R L; Ostrom, N E; Ostrom, P H; Gandhi, H; Breznak, J A

    2003-01-01

    The relative importance of individual microbial pathways in nitrous oxide (N(2)O) production is not well known. The intramolecular distribution of (15)N in N(2)O provides a basis for distinguishing biological pathways. Concentrated cell suspensions of Methylococcus capsulatus Bath and Nitrosomonas europaea were used to investigate the site preference of N(2)O by microbial processes during nitrification. The average site preference of N(2)O formed during hydroxylamine oxidation by M. capsulatus Bath (5.5 +/- 3.5 per thousand) and N. europaea (-2.3 +/- 1.9 per thousand) and nitrite reduction by N. europaea (-8.3 +/- 3.6 per thousand) differed significantly (ANOVA, f((2,35)) = 247.9, p = 0). These results demonstrate that the mechanisms for hydroxylamine oxidation are distinct in M. capsulatus Bath and N. europaea. The average delta(18)O-N(2)O values of N(2)O formed during hydroxylamine oxidation for M. capsulatus Bath (53.1 +/- 2.9 per thousand) and N. europaea (-23.4 +/- 7.2 per thousand) and nitrite reduction by N. europaea (4.6 +/- 1.4 per thousand) were significantly different (ANOVA, f((2,35)) = 279.98, p = 0). Although the nitrogen isotope value of the substrate, hydroxylamine, was similar in both cultures, the observed fractionation (delta(15)N) associated with N(2)O production via hydroxylamine oxidation by M. capsulatus Bath and N. europaea (-2.3 and 26.0 per thousand, respectively) provided evidence that differences in isotopic fractionation were associated with these two organisms. The site preferences in this study are the first measured values for isolated microbial processes. The differences in site preference are significant and indicate that isotopomers provide a basis for apportioning biological processes producing N(2)O. PMID:12661029

  4. Transcription of nitrification genes by the methane-oxidizing bacterium, Methylococcus capsulatus strain Bath.

    PubMed

    Poret-Peterson, Amisha T; Graham, James E; Gulledge, Jay; Klotz, Martin G

    2008-12-01

    Methylococcus capsulatus strain Bath, a methane-oxidizing bacterium, and ammonia-oxidizing bacteria (AOB) carry out the first step of nitrification, the oxidation of ammonia to nitrite, through the intermediate hydroxylamine. AOB use hydroxylamine oxidoreductase (HAO) to produce nitrite. M. capsulatus Bath was thought to oxidize hydroxylamine with cytochrome P460 (cytL), until the recent discovery of an hao gene in its genome. We used quantitative PCR analyses of cDNA from M. capsulatus Bath incubated with CH(4) or CH(4) plus 5 mM (NH(4))(2)SO(4) to determine whether cytL and hao transcript levels change in response to ammonia. While mRNA levels for cytL were not affected by ammonia, hao mRNA levels increased by 14.5- and 31-fold in duplicate samples when a promoter proximal region of the transcript was analyzed, and by sixfold when a region at the distal end of the transcript was analyzed. A conserved open reading frame, orf2, located 3' of hao in all known AOB genomes and in M. capsulatus Bath, was cotranscribed with hao and showed increased mRNA levels in the presence of ammonia. These data led to designating this gene pair as haoAB, with the role of haoB still undefined. We also determined mRNA levels for additional genes that encode proteins involved in N-oxide detoxification: cytochrome c'-beta (CytS) and nitric oxide (NO) reductase (NorCB). Whereas cytS mRNA levels increased in duplicate samples by 28.5- and 40-fold in response to ammonia, the cotranscribed norC-norB mRNA did not increase. Our results strongly suggest that M. capsulatus Bath possesses a functional, ammonia-responsive HAO involved in nitrification. PMID:18650926

  5. Characterization of recombinant fructose-1,6-bisphosphate aldolase from Methylococcus capsulatus Bath.

    PubMed

    Rozova, O N; Khmelenina, V N; Mustakhimov, I I; Reshetnikov, A S; Trotsenko, Y A

    2010-07-01

    The gene fba from the thermotolerant obligate methanotroph Methylococcus capsulatus Bath was cloned and expressed in Escherichia coli BL21(DE3). The fructose-1,6-bisphosphate aldolase (FBA) carrying six His on the C-end was purified by affinity metal chelating chromatography. The Mc. capsulatus FBA is a hexameric enzyme (240 kDa) that is activated by Co2+ and inhibited by EDTA. The enzyme displays low K(m) to fructose-1,6-bisphosphate (FBP) and higher K(m) to the substrates of aldol condensation, dihydroxyacetone phosphate and glyceraldehyde-3-phosphate. The FBA also catalyzes sedoheptulose-1,7-bisphosphate cleavage. The presence of Co2+ in the reaction mixture changes the kinetics of FBP hydrolysis and is accompanied by inhibition of the reaction by 2 mM FBP. Phylogenetically, the Mc. capsulatus enzyme belongs to the type B of class II FBAs showing high identity of translated amino acid sequence with FBAs from autotrophic bacteria. The role of the FBA in metabolism of Mc. capsulatus Bath, which realizes simultaneously three C(1) assimilating pathways (the ribulose monophosphate, the ribulose bisphosphate, and the serine cycles), is discussed. PMID:20673213

  6. Heterotrophic bacteria growing in association with Methylococcus capsulatus (Bath) in a single cell protein production process.

    PubMed

    Bothe, Harald; Møller Jensen, K; Mergel, A; Larsen, J; Jørgensen, C; Bothe, Hermann; Jørgensen, L

    2002-06-01

    The methanotrophic bacterium Methylococcus capsulatus (Bath) grows on pure methane. However, in a single cell protein production process using natural gas as methane source, a bacterial consortium is necessary to support growth over longer periods in continuous cultures. In different bioreactors of Norferm Danmark A/S, three bacteria consistently invaded M. capsulatus cultures growing under semi-sterile conditions in continuous culture. These bacteria have now been identified as a not yet described member of the Aneurinibacillus group, a Brevibacillus agri strain, and an acetate-oxidiser of the genus Ralstonia. The physiological roles of these bacteria in the bioreactor culture growing on natural, non-pure methane gas are discussed. The heterotrophic bacteria do not have the genetic capability to produce either the haemolytic enterotoxin complex HBL or non-haemolytic enterotoxin. PMID:12073128

  7. Characterization of the pyrophosphate-dependent 6-phosphofructokinase from Methylococcus capsulatus Bath.

    PubMed

    Reshetnikov, Alexander S; Rozova, Olga N; Khmelenina, Valentina N; Mustakhimov, Ildar I; Beschastny, Alexander P; Murrell, J Colin; Trotsenko, Yuri A

    2008-11-01

    An active pyrophosphate-dependent 6-phosphofructokinase (PPi-PFK) from the thermotolerant methanotroph Methylococcus capsulatus Bath, containing a six-residue polyhistidine tag, was characterized. The enzyme was homodimeric (2 x 45 kDa), nonallosteric and most active at pH 7.0. PPi-PFK catalyzed reactions of PPi-dependent phosphorylation of fructose-6-phosphate (F-6-P) (K(m) 2.27 mM and V(max) 7.6 U mg(-1) of protein), sedoheptulose-7-phosphate (K(m) 0.027 mM and V(max) 31 U mg(-1)) and ribulose-5-phosphate. In the reaction with F-6-P, the apparent K(m) for PPi was 0.027 mM, while in the reverse reaction, K(m) for orthophosphate was 8.69 mM and that for fructose-1,6-bisphosphate 0.328 mM (V(max) 9.0 U mg(-1)). Phylogenetically, M. capsulatus PPi-PFK was most similar to PPi-PFKs from the lithoautotrophic ammonia oxidizers Nitrosomonas europaea (74.0%), Nitrosospira multiformis (73.6%) and Betaproteobacterial methylotroph Methylibium petroleiphilum PM1 (71.6% identity). Genes coding PPi-PFK and a putative V-type H(+)-translocating pyrophosphatase (H(+)-PPi-ase) were cotranscribed as an operon. The potential significance of the PPi-PFK for regulation of carbon and energy fluxes in M. capsulatus Bath is discussed. PMID:19054082

  8. Analysing the outer membrane subproteome of Methylococcus capsulatus (Bath) using proteomics and novel biocomputing tools.

    PubMed

    Berven, Frode S; Karlsen, Odd André; Straume, Anne Hege; Flikka, Kristian; Murrell, J Colin; Fjellbirkeland, Anne; Lillehaug, Johan R; Eidhammer, Ingvar; Jensen, Harald B

    2006-02-01

    High-resolution two-dimensional gel electrophoresis and mass spectrometry has been used to identify the outer membrane (OM) subproteome of the Gram-negative bacterium Methylococcus capsulatus (Bath). Twenty-eight unique polypeptide sequences were identified from protein samples enriched in OMs. Only six of these polypeptides had previously been identified. The predictions from novel bioinformatic methods predicting beta-barrel outer membrane proteins (OMPs) and OM lipoproteins were compared to proteins identified experimentally. BOMP ( http://www.bioinfo.no/tools/bomp ) predicted 43 beta-barrel OMPs (1.45%) from the 2,959 annotated open reading frames. This was a lower percentage than predicted from other Gram-negative proteomes (1.8-3%). More than half of the predicted BOMPs in M. capsulatus were annotated as (conserved) hypothetical proteins with significant similarity to very few sequences in Swiss-Prot or TrEMBL. The experimental data and the computer predictions indicated that the protein composition of the M. capsulatus OM subproteome was different from that of other Gram-negative bacteria studied in a similar manner. A new program, Lipo, was developed that can analyse entire predicted proteomes and give a list of recognised lipoproteins categorised according to their lipo-box similarity to known Gram-negative lipoproteins ( http://www.bioinfo.no/tools/lipo ). This report is the first using a proteomics and bioinformatics approach to identify the OM subproteome of an obligate methanotroph. PMID:16311759

  9. Genomic insights into methanotrophy: the complete genome sequence of Methylococcus capsulatus (Bath).

    PubMed

    Ward, Naomi; Larsen, Øivind; Sakwa, James; Bruseth, Live; Khouri, Hoda; Durkin, A Scott; Dimitrov, George; Jiang, Lingxia; Scanlan, David; Kang, Katherine H; Lewis, Matt; Nelson, Karen E; Methé, Barbara; Wu, Martin; Heidelberg, John F; Paulsen, Ian T; Fouts, Derrick; Ravel, Jacques; Tettelin, Hervé; Ren, Qinghu; Read, Tim; DeBoy, Robert T; Seshadri, Rekha; Salzberg, Steven L; Jensen, Harald B; Birkeland, Nils Kåre; Nelson, William C; Dodson, Robert J; Grindhaug, Svenn H; Holt, Ingeborg; Eidhammer, Ingvar; Jonasen, Inge; Vanaken, Susan; Utterback, Terry; Feldblyum, Tamara V; Fraser, Claire M; Lillehaug, Johan R; Eisen, Jonathan A

    2004-10-01

    Methanotrophs are ubiquitous bacteria that can use the greenhouse gas methane as a sole carbon and energy source for growth, thus playing major roles in global carbon cycles, and in particular, substantially reducing emissions of biologically generated methane to the atmosphere. Despite their importance, and in contrast to organisms that play roles in other major parts of the carbon cycle such as photosynthesis, no genome-level studies have been published on the biology of methanotrophs. We report the first complete genome sequence to our knowledge from an obligate methanotroph, Methylococcus capsulatus (Bath), obtained by the shotgun sequencing approach. Analysis revealed a 3.3-Mb genome highly specialized for a methanotrophic lifestyle, including redundant pathways predicted to be involved in methanotrophy and duplicated genes for essential enzymes such as the methane monooxygenases. We used phylogenomic analysis, gene order information, and comparative analysis with the partially sequenced methylotroph Methylobacterium extorquens to detect genes of unknown function likely to be involved in methanotrophy and methylotrophy. Genome analysis suggests the ability of M. capsulatus to scavenge copper (including a previously unreported nonribosomal peptide synthetase) and to use copper in regulation of methanotrophy, but the exact regulatory mechanisms remain unclear. One of the most surprising outcomes of the project is evidence suggesting the existence of previously unsuspected metabolic flexibility in M. capsulatus, including an ability to grow on sugars, oxidize chemolithotrophic hydrogen and sulfur, and live under reduced oxygen tension, all of which have implications for methanotroph ecology. The availability of the complete genome of M. capsulatus (Bath) deepens our understanding of methanotroph biology and its relationship to global carbon cycles. We have gained evidence for greater metabolic flexibility than was previously known, and for genetic components that

  10. Genomic Insights into Methanotrophy: The Complete Genome Sequence of Methylococcus capsulatus (Bath)

    PubMed Central

    2004-01-01

    Methanotrophs are ubiquitous bacteria that can use the greenhouse gas methane as a sole carbon and energy source for growth, thus playing major roles in global carbon cycles, and in particular, substantially reducing emissions of biologically generated methane to the atmosphere. Despite their importance, and in contrast to organisms that play roles in other major parts of the carbon cycle such as photosynthesis, no genome-level studies have been published on the biology of methanotrophs. We report the first complete genome sequence to our knowledge from an obligate methanotroph, Methylococcus capsulatus (Bath), obtained by the shotgun sequencing approach. Analysis revealed a 3.3-Mb genome highly specialized for a methanotrophic lifestyle, including redundant pathways predicted to be involved in methanotrophy and duplicated genes for essential enzymes such as the methane monooxygenases. We used phylogenomic analysis, gene order information, and comparative analysis with the partially sequenced methylotroph Methylobacterium extorquens to detect genes of unknown function likely to be involved in methanotrophy and methylotrophy. Genome analysis suggests the ability of M. capsulatus to scavenge copper (including a previously unreported nonribosomal peptide synthetase) and to use copper in regulation of methanotrophy, but the exact regulatory mechanisms remain unclear. One of the most surprising outcomes of the project is evidence suggesting the existence of previously unsuspected metabolic flexibility in M. capsulatus, including an ability to grow on sugars, oxidize chemolithotrophic hydrogen and sulfur, and live under reduced oxygen tension, all of which have implications for methanotroph ecology. The availability of the complete genome of M. capsulatus (Bath) deepens our understanding of methanotroph biology and its relationship to global carbon cycles. We have gained evidence for greater metabolic flexibility than was previously known, and for genetic components that

  11. Fourier transform infrared characterization of the azido complex of methane monooxygenase hydroxylase from Methylococcus capsulatus (Bath).

    PubMed

    Lu, Shen; Sazinsky, Matthew H; Whittaker, James W; Lippard, Stephen J; Moënne-Loccoz, Pierre

    2005-03-30

    The azido complex formed in oxidized methane monooxygenase from Methylococcus capsulatus (Bath) was investigated with resonance Raman and FTIR techniques. These experiments show the presence of a nuas(NNN) at approximately 2077 cm-1 which splits to two components at 2059 and 2073 cm-1 with 15N14N2. The vibrational data are assigned to an azido complex bound terminally to one iron(III) at the diiron center. When the azido complex is illuminated at 15 K, a new nuas(NNN) is observed at 2136 cm-1 which is assigned to a photodissociated HN3 within the substrate pocket. We propose a model where an aqua ligand engages a hydrogen bond interaction with the 1N atom of the azido group and acts as at a proton donor during the photolysis process. PMID:15783178

  12. Oxidation of hydroxylamine by cytochrome P-460 of the obligate methylotroph Methylococcus capsulatus Bath.

    PubMed Central

    Zahn, J A; Duncan, C; DiSpirito, A A

    1994-01-01

    An enzyme capable of the oxidation of hydroxylamine to nitrite was isolated from the obligate methylotroph Methylococcus capsulatus Bath. The absorption spectra in cell extracts, electron paramagnetic resonance spectra, molecular weight, covalent attachment of heme group to polypeptide, and enzymatic activities suggest that the enzyme is similar to cytochrome P-460, a novel iron-containing protein previously observed only in Nitrosomonas europaea. The native and subunit molecular masses of the M. capsulatus Bath protein were 38,900 and 16,390 Da, respectively; the isoelectric point was 6.98. The enzyme has approximately one iron and one copper atom per subunit. The electron paramagnetic resonance spectrum of the protein showed evidence for a high-spin ferric heme. In contrast to the enzyme from N. europaea, a 13-nm blue shift in the soret band of the ferrocytochrome (463 nm in cell extracts to 450 nm in the final sample) occurred during purification. The amino acid composition and N-terminal amino acid sequence of the enzyme from M. capsulatus Bath was similar but not identical to those of cytochrome P-460 of N. europaea. In cell extracts, the identity of the biological electron acceptor is as yet unestablished. Cytochrome c-555 is able to accept electrons from cytochrome P-460, although the purified enzyme required phenazine methosulfate for maximum hydroxylamine oxidation activity (specific activity, 366 mol of O2 per s per mol of enzyme). Hydroxylamine oxidation rates were stimulated approximately 2-fold by 1 mM cyanide and 1.5-fold by 0.1 mM 8-hydroxyquinoline. Images PMID:7928947

  13. Quantitative proteomic analysis of metabolic regulation by copper ions in Methylococcus capsulatus (Bath).

    PubMed

    Kao, Wei-Chun; Chen, Yet-Ran; Yi, Eugene C; Lee, Hookeun; Tian, Qiang; Wu, Keh-Ming; Tsai, Shih-Feng; Yu, Steve S-F; Chen, Yu-Ju; Aebersold, Ruedi; Chan, Sunney I

    2004-12-01

    Copper ions switch the oxidation of methane by soluble methane monooxygenase to particulate methane monooxygenase in Methylococcus capsulatus (Bath). Toward understanding the change in cellular metabolism related to this transcriptional and metabolic switch, we have undertaken genomic sequencing and quantitative comparative analysis of the proteome in M. capsulatus (Bath) grown under different copper-to-biomass ratios by cleavable isotope-coded affinity tag technology. Of the 682 proteins identified, the expressions of 60 proteins were stimulated by at least 2-fold by copper ions; 68 proteins were down-regulated by 2-fold or more. The 60 proteins overexpressed included the methane and carbohydrate metabolic enzymes, while the 68 proteins suppressed were mainly responsible for cellular signaling processes, indicating a role of copper ions in the expression of the genes associated with the metabolism of the organism downstream of methane oxidation. The study has also provided a complete map of the C1 metabolism pathways in this methanotroph and clarified the interrelationships between them. PMID:15385566

  14. Characterization of a prokaryotic haemerythrin from the methanotrophic bacterium Methylococcus capsulatus (Bath).

    PubMed

    Karlsen, Odd A; Ramsevik, Linda; Bruseth, Live J; Larsen, Øivind; Brenner, Annette; Berven, Frode S; Jensen, Harald B; Lillehaug, Johan R

    2005-05-01

    For a long time, the haemerythrin family of proteins was considered to be restricted to only a few phyla of marine invertebrates. When analysing differential protein expression in the methane-oxidizing bacterium, Methylococcus capsulatus (Bath), grown at a high and low copper-to-biomass ratio, respectively, we identified a putative prokaryotic haemerythrin expressed in high-copper cultures. Haemerythrins are recognized by a conserved sequence motif that provides five histidines and two carboxylate ligands which coordinate two iron atoms. The diiron site is located in a hydrophobic pocket and is capable of binding O(2). We cloned the M. capsulatus haemerythrin gene and expressed it in Escherichia coli as a fusion protein with NusA. The haemerythrin protein was purified to homogeneity cleaved from its fusion partner. Recombinant M. capsulatus haemerythrin (McHr) was found to fold into a stable protein. Sequence similarity analysis identified all the candidate residues involved in the binding of diiron (His22, His58, Glu62, His77, His81, His117, Asp122) and the amino acids forming the hydrophobic pocket in which O(2) may bind (Ile25, Phe59, Trp113, Leu114, Ile118). We were also able to model a three-dimensional structure of McHr maintaining the correct positioning of these residues. Furthermore, UV/vis spectrophotometric analysis demonstrated the presence of conjugated diiron atoms in McHr. A comprehensive genomic database search revealed 21 different prokaryotes containing the haemerythrin signature (PROSITE 00550), indicating that these putative haemerythrins may be a conserved prokaryotic subfamily. PMID:15885093

  15. Transcriptomic profiling of Methylococcus capsulatus (Bath) during growth with two different methane monooxygenases.

    PubMed

    Larsen, Øivind; Karlsen, Odd A

    2016-04-01

    Methylococcus capsulatus (Bath) is a methanotroph that possesses both a membrane-embedded (pMMO) and a soluble methane monooxygenase (sMMO). The expression of these two MMO's is tightly controlled by the availability of copper in the growth medium, but the underlying mechanisms and the number of genes involved in this switch in methane oxidation is not yet fully elucidated. Microarray analyses were used to assess the transcriptome in cells producing either pMMO or sMMO. A total of 137 genes were differentially expressed, with 87 genes showing a significant up-regulation during sMMO production. The majority of the differentially expressed genes could be assigned to functional roles in the energy metabolism and transport. Furthermore, three copper responding gene clusters were discovered, including an extended cluster that also harbors the genes for sMMO. Our data also indicates that major changes takes place in the respiratory chain between pMMO- and sMMO-producing cells, and that quinone are predominantly used as the electron donors for methane oxidation by pMMO. Intriguingly, a large proportion of the differentially expressed genes between pMMO- and sMMO-producing cells encode c-type cytochromes. By combining microarray- and mass spectrometry data, a total of 35 c-type cytochromes are apparently expressed in M. capsulatus when grown in nitrate mineral salt medium with methane as sole energy and carbon source, and the expression of 21 of these respond to the availability of copper. Interestingly, several of these c-type cytochromes are recovered from the cell surface, suggesting that extracellular electron transfers may occur in M. capsulatus. PMID:26687591

  16. High-molecular-mass multi-c-heme cytochromes from Methylococcus capsulatus bath.

    PubMed

    Bergmann, D J; Zahn, J A; DiSpirito, A A

    1999-02-01

    The polypeptide and structural gene for a high-molecular-mass c-type cytochrome, cytochrome c553O, was isolated from the methanotroph Methylococcus capsulatus Bath. Cytochrome c553O is a homodimer with a subunit molecular mass of 124,350 Da and an isoelectric point of 6. 0. The heme c concentration was estimated to be 8.2 +/- 0.4 mol of heme c per subunit. The electron paramagnetic resonance spectrum showed the presence of multiple low spin, S = 1/2, hemes. A degenerate oligonucleotide probe synthesized based on the N-terminal amino acid sequence of cytochrome c553O was used to identify a DNA fragment from M. capsulatus Bath that contains occ, the gene encoding cytochrome c553O. occ is part of a gene cluster which contains three other open reading frames (ORFs). ORF1 encodes a putative periplasmic c-type cytochrome with a molecular mass of 118, 620 Da that shows approximately 40% amino acid sequence identity with occ and contains nine c-heme-binding motifs. ORF3 encodes a putative periplasmic c-type cytochrome with a molecular mass of 94, 000 Da and contains seven c-heme-binding motifs but shows no sequence homology to occ or ORF1. ORF4 encodes a putative 11,100-Da protein. The four ORFs have no apparent similarity to any proteins in the GenBank database. The subunit molecular masses, arrangement and number of hemes, and amino acid sequences demonstrate that cytochrome c553O and the gene products of ORF1 and ORF3 constitute a new class of c-type cytochrome. PMID:9922265

  17. The Methylococcus capsulatus (Bath) Secreted Protein, MopE*, Binds Both Reduced and Oxidized Copper

    PubMed Central

    Ve, Thomas; Mathisen, Karina; Helland, Ronny; Karlsen, Odd A.; Fjellbirkeland, Anne; Røhr, Åsmund K.; Andersson, K. Kristoffer; Pedersen, Rolf-Birger; Lillehaug, Johan R.; Jensen, Harald B.

    2012-01-01

    Under copper limiting growth conditions the methanotrophic bacterium Methylococcus capsulatus (Bath) secrets essentially only one protein, MopE*, to the medium. MopE* is a copper-binding protein whose structure has been determined by X-ray crystallography. The structure of MopE* revealed a unique high affinity copper binding site consisting of two histidine imidazoles and one kynurenine, the latter an oxidation product of Trp130. In this study, we demonstrate that the copper ion coordinated by this strong binding site is in the Cu(I) state when MopE* is isolated from the growth medium of M. capsulatus. The conclusion is based on X-ray Near Edge Absorption spectroscopy (XANES), and Electron Paramagnetic Resonance (EPR) studies. EPR analyses demonstrated that MopE*, in addition to the strong copper-binding site, also binds Cu(II) at two weaker binding sites. Both Cu(II) binding sites have properties typical of non-blue type II Cu (II) centres, and the strongest of the two Cu(II) sites is characterised by a relative high hyperfine coupling of copper (A|| = 20 mT). Immobilized metal affinity chromatography binding studies suggests that residues in the N-terminal part of MopE* are involved in forming binding site(s) for Cu(II) ions. Our results support the hypothesis that MopE plays an important role in copper uptake, possibly making use of both its high (Cu(I) and low Cu(II) affinity properties. PMID:22916218

  18. The Methylococcus capsulatus (Bath) secreted protein, MopE*, binds both reduced and oxidized copper.

    PubMed

    Ve, Thomas; Mathisen, Karina; Helland, Ronny; Karlsen, Odd A; Fjellbirkeland, Anne; Røhr, Åsmund K; Andersson, K Kristoffer; Pedersen, Rolf-Birger; Lillehaug, Johan R; Jensen, Harald B

    2012-01-01

    Under copper limiting growth conditions the methanotrophic bacterium Methylococcus capsulatus (Bath) secrets essentially only one protein, MopE*, to the medium. MopE* is a copper-binding protein whose structure has been determined by X-ray crystallography. The structure of MopE* revealed a unique high affinity copper binding site consisting of two histidine imidazoles and one kynurenine, the latter an oxidation product of Trp130. In this study, we demonstrate that the copper ion coordinated by this strong binding site is in the Cu(I) state when MopE* is isolated from the growth medium of M. capsulatus. The conclusion is based on X-ray Near Edge Absorption spectroscopy (XANES), and Electron Paramagnetic Resonance (EPR) studies. EPR analyses demonstrated that MopE*, in addition to the strong copper-binding site, also binds Cu(II) at two weaker binding sites. Both Cu(II) binding sites have properties typical of non-blue type II Cu (II) centres, and the strongest of the two Cu(II) sites is characterised by a relative high hyperfine coupling of copper (A(||) =20 mT). Immobilized metal affinity chromatography binding studies suggests that residues in the N-terminal part of MopE* are involved in forming binding site(s) for Cu(II) ions. Our results support the hypothesis that MopE plays an important role in copper uptake, possibly making use of both its high (Cu(I) and low Cu(II) affinity properties. PMID:22916218

  19. Structure of the redox sensor domain of Methylococcus capsulatus (Bath) MmoS‡

    PubMed Central

    Ukaegbu, Uchechi E.; Rosenzweig, Amy C.

    2009-01-01

    MmoS from Methylococcus capsulatus (Bath) is the multidomain sensor protein of a two component signaling system proposed to play a role in the copper-mediated regulation of soluble methane monooxygenase (sMMO). MmoS binds an FAD cofactor within its N-terminal tandem Per-Arnt-Sim (PAS) domains, suggesting that it functions as a redox sensor. The crystal structure of the MmoS tandem PAS domains, designated PAS-A and PAS-B, has been determined to 2.34 Å resolution. Both domains adopt the typical PAS domain α/β topology and are structurally similar. The two domains are linked by a long α helix and do not interact with one another. The FAD cofactor is housed solely within PAS-A and is stabilized by an extended hydrogen bonding network. The overall fold of PAS-A is similar to other flavin-containing PAS domains, but homodimeric interactions in other structures are not observed in the MmoS sensor, which crystallized as a monomer. The structure both provides new insight into the architecture of tandem PAS domains and suggests specific residues that may play a role in MmoS FAD redox chemistry and subsequent signal transduction. PMID:19271777

  20. Steady-state kinetic analysis of soluble methane mono-oxygenase from Methylococcus capsulatus (Bath).

    PubMed Central

    Green, J; Dalton, H

    1986-01-01

    A steady-state kinetic analysis of purified soluble methane mono-oxygenase of Methylococcus capsulatus (Bath) was performed. The enzyme was found to follow a concerted-substitution mechanism. Methane binds to the enzyme followed by NADH, which reacts to yield reduced enzyme and NAD+. The reduced enzyme-methane complex binds O2 to give a second ternary complex, which breaks down to release water and methanol. In this way the enzyme can control the supply of electrons to the active site to coincide with the arrival of methane. Product-inhibition studies (with propylene as substrate) supported the reaction mechanism proposed. Ki values for NAD+ and propylene oxide are reported. The Km for NADH varied from 25 microM to 300 microM, depending on the nature of the hydrocarbon substrate, and thus supports the proposed reaction sequence. With methane as substrate the Km values for methane, NADH and O2 were shown to be 3 microM, 55.8 microM and 16.8 microM respectively. With propylene as substrate the Km values for propylene, NADH and O2 were 0.94 microM, 25.2 microM and 12.7-15.9 microM respectively. Methane mono-oxygenase was shown to be well adapted to the oxidation of methane compared with other straight-chain alkanes. PMID:3098230

  1. Oxidation of ultrafast radical clock substrate probes by the soluble methane monooxygenase from Methylococcus capsulatus (Bath).

    PubMed

    Valentine, A M; LeTadic-Biadatti, M H; Toy, P H; Newcomb, M; Lippard, S J

    1999-04-16

    Radical clock substrate probes were used to assess the viability of a discrete substrate radical species in the mechanism of hydrocarbon oxidation by the soluble methane monooxygenase (sMMO) from Methylococcus capsulatus (Bath). New substituted cyclopropane probes were used with very fast ring-opening rate constants and other desirable attributes, such as the ability to discriminate between radical and cationic intermediates. Oxidation of these substrates by a reconstituted sMMO system resulted in no rearranged products, allowing an upper limit of 150 fs to be placed on the lifetime of a putative radical species. This limit strongly suggests that there is no such substrate radical intermediate. The two enantiomers of trans-1-methyl-2-phenyl-cyclopropane were prepared, and the regioselectivity of their oxidation to the corresponding cyclopropylmethanol and cyclopropylphenol products was determined. The results are consistent with selective orientation of the two enantiomeric substrates in the hydrophobic cavity at the active site of sMMO, specific models for which were examined by molecular modeling. PMID:10196150

  2. NMR structure of the flavin domain from soluble methane monooxygenase reductase from Methylococcus capsulatus (Bath).

    PubMed

    Chatwood, Lisa L; Müller, Jens; Gross, John D; Wagner, Gerhard; Lippard, Stephen J

    2004-09-28

    Soluble methane monooxygenase (sMMO) catalyzes the hydroxylation of methane by dioxygen to methanol, the first step in carbon assimilation by methanotrophs. This multicomponent system transfers electrons from NADH through a reductase component to the non-heme diiron center in the hydroxylase where O(2) is activated. The reductase component comprises three distinct domains, a [2Fe-2S] ferredoxin domain along with FAD- and NADH-binding domains. We report the solution structure of the reduced 27.6 kDa FAD- and NADH-binding domains (MMOR-FAD) of the reductase from Methylococcus capsulatus (Bath). The FAD-binding domain consists of a six-stranded antiparallel beta-barrel and one alpha-helix, with the first 10 N-terminal residues unstructured. In the interface between the two domains, the FAD cofactor is tightly bound in an unprecedented extended conformation. The NADH-binding domain consists of a five-stranded parallel beta-sheet with four alpha-helices packing closely around this sheet. MMOR-FAD is structurally homologous to other FAD-containing oxidoreductases, and we expect similar structures for the FAD/NADH-binding domains of reductases that occur in other multicomponent monooxygenases. PMID:15379538

  3. Formaldehyde dehydrogenase preparations from Methylococcus capsulatus (Bath) comprise methanol dehydrogenase and methylene tetrahydromethanopterin dehydrogenase.

    PubMed

    Adeosun, Ekundayo K; Smith, Thomas J; Hoberg, Anne-Mette; Velarde, Giles; Ford, Robert; Dalton, Howard

    2004-03-01

    In methylotrophic bacteria, formaldehyde is an important but potentially toxic metabolic intermediate that can be assimilated into biomass or oxidized to yield energy. Previously reported was the purification of an NAD(P)(+)-dependent formaldehyde dehydrogenase (FDH) from the obligate methane-oxidizing methylotroph Methylococcus capsulatus (Bath), presumably important in formaldehyde oxidation, which required a heat-stable factor (known as the modifin) for FDH activity. Here, the major protein component of this FDH preparation was shown by biophysical techniques to comprise subunits of 64 and 8 kDa in an alpha(2)beta(2) arrangement. N-terminal sequencing of the subunits of FDH, together with enzymological characterization, showed that the alpha(2)beta(2) tetramer was a quinoprotein methanol dehydrogenase of the type found in other methylotrophs. The FDH preparations were shown to contain a highly active NAD(P)(+)-dependent methylene tetrahydromethanopterin dehydrogenase that was the probable source of the NAD(P)(+)-dependent formaldehyde oxidation activity. These results support previous findings that methylotrophs possess multiple pathways for formaldehyde dissimilation. PMID:14993320

  4. Presence of methyl sterol and bacteriohopanepolyol in an outer-membrane preparation from Methylococcus capsulatus (Bath)

    NASA Technical Reports Server (NTRS)

    Jahnke, Linda L.; Stan-Lotter, Helga; Kato, Katharine; Hochstein, Lawrence I.

    1992-01-01

    Cytoplasmic/intracytoplasmic and outer membrane preparations of Methylococcus capsulatus (Bath) were isolated by sucrose density gradient centrifugation of a total membrane fraction prepared by disruption using a French pressure cell. The cytoplasmic and/or intracytoplasmic membrane fraction consisted of two distinct bands, Ia and Ib (buoyant densities 1.16 and 1.18 g ml (exp -1), respectively) that together contained 57% of the protein, 68% of the phospholipid, 73% of the ubiquinone and 89% of the CN-sensitive NADH oxidase activity. The only apparent difference between these two cytoplasmic bands was a much higher phospholipid content for Ia. The outer membrane fraction (buoyant density 1.23-1.24 g ml (exp -1)) contained 60% of the lipopolysaccharide-associated, beta-hydroxypalmitic acid, 74% of the methylsterol, and 66% of the bacteriohopanepolyol (BHP); phospholipid to methyl sterol or BHP ratios were 6:1. Methanol dehydrogenase activity and a c-type cytochrome were also present in this outer membrane fraction. Phospholipase A activity was present in borh the cytoplasmic membrane and outer membrane fractions. The unique distribution of cyclic triterpenes may reflect a specific role in conferring outer membrane stability in this methanotrophic bacterium.

  5. The bacteriohemerythrin from Methylococcus capsulatus (Bath): Crystal structures reveal that Leu114 regulates a water tunnel.

    PubMed

    Chen, Kelvin H-C; Chuankhayan, Phimonphan; Wu, Hsin-Hui; Chen, Chun-Jung; Fukuda, Mitsuhiro; Yu, Steve S-F; Chan, Sunney I

    2015-09-01

    The bacteriohemerythrin (McHr) from Methylococcus capsulatus (Bath) is an oxygen carrier that serves as a transporter to deliver O2 from the cytosol of the bacterial cell body to the particulate methane monooxygenase residing in the intracytoplasmic membranes for methane oxidation. Here we report X-ray protein crystal structures of the recombinant wild type (WT) McHr and its L114A, L114Y and L114F mutants. The structure of the WT reveals a possible water tunnel in the McHr that might be linked to its faster autoxidation relative to hemerythrin in marine invertebrates. With Leu114 positioned at the end of this putative water tunnel, the hydrophobic side chain of this residue seems to play a prominent role in controlling the access of the water molecule required for autoxidation. This hypothesis is examined by comparing the autoxidation rates of the WT McHr with those of the L114A, L114Y and L114F mutants. The biochemical data are correlated with structural insights derived from the analysis of the putative water tunnels in the various McHr proteins provided by the X-ray structures. PMID:25890483

  6. Structure of the redox sensor domain of Methylococcus capsulatus (Bath) MmoS.

    PubMed

    Ukaegbu, Uchechi E; Rosenzweig, Amy C

    2009-03-17

    MmoS from Methylococcus capsulatus (Bath) is the multidomain sensor protein of a two-component signaling system proposed to play a role in the copper-mediated regulation of soluble methane monooxygenase (sMMO). MmoS binds an FAD cofactor within its N-terminal tandem Per-Arnt-Sim (PAS) domains, suggesting that it functions as a redox sensor. The crystal structure of the MmoS tandem PAS domains, designated PAS-A and PAS-B, has been determined to 2.34 A resolution. Both domains adopt the typical PAS domain alpha/beta topology and are structurally similar. The two domains are linked by a long alpha helix and do not interact with one another. The FAD cofactor is housed solely within PAS-A and is stabilized by an extended hydrogen bonding network. The overall fold of PAS-A is similar to those of other flavin-containing PAS domains, but homodimeric interactions in other structures are not observed in the MmoS sensor, which crystallized as a monomer. The structure both provides new insight into the architecture of tandem PAS domains and suggests specific residues that may play a role in MmoS FAD redox chemistry and subsequent signal transduction. PMID:19271777

  7. Structure of the Redox Sensor Domain of Methylococcus capsulatus (Bath) MmoS

    SciTech Connect

    Ukaegbu, Uchechi E.; Rosenzweig, Amy C.

    2009-06-01

    MmoS from Methylococcus capsulatus (Bath) is the multidomain sensor protein of a two-component signaling system proposed to play a role in the copper-mediated regulation of soluble methane monooxygenase (sMMO). MmoS binds an FAD cofactor within its N-terminal tandem Per-Arnt-Sim (PAS) domains, suggesting that it functions as a redox sensor. The crystal structure of the MmoS tandem PAS domains, designated PAS-A and PAS-B, has been determined to 2.34 {angstrom} resolution. Both domains adopt the typical PAS domain {alpha}/{beta} topology and are structurally similar. The two domains are linked by a long {alpha} helix and do not interact with one another. The FAD cofactor is housed solely within PAS-A and is stabilized by an extended hydrogen bonding network. The overall fold of PAS-A is similar to those of other flavin-containing PAS domains, but homodimeric interactions in other structures are not observed in the MmoS sensor, which crystallized as a monomer. The structure both provides new insight into the architecture of tandem PAS domains and suggests specific residues that may play a role in MmoS FAD redox chemistry and subsequent signal transduction.

  8. Bacteriohemerythrin bolsters the activity of the particulate methane monooxygenase (pMMO) in Methylococcus capsulatus (Bath).

    PubMed

    Chen, Kelvin H-C; Wu, Hsin-Hui; Ke, Si-Fu; Rao, Ya-Ting; Tu, Chia-Ming; Chen, Yu-Ping; Kuei, Kuo-Hsuan; Chen, Ying-Siao; Wang, Vincent C-C; Kao, Wei-Chun; Chan, Sunney I

    2012-06-01

    Recently, a native bacteriohemerythrin (McHr) has been identified in Methylococcus capsulatus (Bath). Both the particulate methane monooxygenase (pMMO) and McHr are over-expressed in cells of this bacterium when this strain of methanotroph is cultured and grown under high copper to biomass conditions. It has been suggested that the role of the McHr is to provide a shuttle to transport dioxygen from the cytoplasm of the cell to the intra-cytoplasmic membranes for consumption by the pMMO. Indeed, McHr enhances the activity of the pMMO when pMMO-enriched membranes are used to assay the enzyme activity. We find that McHr can dramatically improve the activity of pMMO toward the epoxidation of propylene to propylene oxide. The maximum activity is observed at a pMMO to McHr concentration ratio of 4:1, where we have obtained specific activities of 103.7nmol propylene oxide/min/mg protein and 122.8nmol propylene oxide/min/mg protein at 45°C when the turnover is driven by NADH and duroquinol, respectively. These results are consistent with the suggestion that the bacterium requires McHr to deliver dioxygen to the pMMO in the intra-cytoplasmic membranes to accomplish efficient catalysis of methane oxidation when the enzyme is over-expressed in the cells. PMID:22484247

  9. Detection and localization of two hydrogenases in Methylococcus capsulatus (Bath) and their potential role in methane metabolism.

    PubMed

    Hanczár, Tímea; Csáki, Robert; Bodrossy, Levente; Murrell, J Colin; Kovács, Kornél L

    2002-02-01

    Methylococcus capsulatus (Bath) was shown to contain two distinct hydrogenases, a soluble hydrogenase and a membrane-bound hydrogenase. This is the first report of a membrane-bound hydrogenase in methanotrophs. Both enzymes were expressed apparently constitutively under normal growth conditions. The soluble hydrogenase was capable of reducing NAD(+) with molecular hydrogen. The activities of both soluble and particulate methane monooxygenases could be driven by molecular hydrogen. This confirmed that molecular hydrogen could be used as a source of reducing power for methane oxidation. Hydrogen-driven methane monooxygenase activities tolerated elevated temperatures and moderate oxygen concentrations. The significance of these findings for biotechnological applications of methanotrophs is discussed. PMID:11807566

  10. Domain engineering of the reductase component of soluble methane monooxygenase from Methylococcus capsulatus (Bath).

    PubMed

    Blazyk, Jessica L; Lippard, Stephen J

    2004-02-13

    Soluble methane monooxygenase (sMMO) from Methylococcus capsulatus (Bath) is a three-component enzyme system that catalyzes the conversion of methane to methanol. A reductase (MMOR), which contains [2Fe-2S] and FAD cofactors, facilitates electron transfer from NADH to the hydroxylase diiron active sites where dioxygen activation and substrate hydroxylation take place. By separately expressing the ferredoxin (MMORFd, MMOR residues 1-98) and FAD/NADH (MMOR-FAD, MMOR residues 99-348) domains of the reductase, nearly all biochemical properties of full-length MMOR are retained, except for interdomain electron transfer rates. To investigate the extent to which rapid electron transfer between domains might be restored and further to explore the modularity of MMOR, MMOR-Fd and MMOR-FAD were connected in a non-native fashion. Four different linker sequences were employed to create MMOR reversed-domain (MMOR-RD) constructs, MMOR(99-342)-linker-MMOR(2-98), with a domain connectivity observed in other homologous oxidoreductases. The optical, redox, and electron transfer properties of the four MMOR-RD proteins were characterized and compared with those of wild-type MMOR. The linker sequence plays a key role in controlling solvent accessibility to the FAD cofactor, as evidenced by perturbed flavin optical spectra, decreased FADox/FADsq redox potentials, and increased steady-state oxidase activities in three of the constructs. Stopped-flow optical spectroscopy revealed slow interdomain electron transfer (k < 0.04 s(-1) at 4 degrees C, compared with 90 s(-1) for wild-type MMOR) for all three MMOR-RD proteins with 7-residue linkers. A long (14-residue), flexible linker afforded much faster electron transfer between the FAD and [2Fe-2S] cofactors (k = 0.9 s(-1) at 4 degrees C). PMID:14613937

  11. Component interactions in the soluble methane monooxygenase system from Methylococcus capsulatus (Bath).

    PubMed

    Gassner, G T; Lippard, S J

    1999-09-28

    The soluble methane monooxygenase system of Methylococcus capsulatus (Bath) includes three protein components: a 251-kDa non-heme dinuclear iron hydroxylase (MMOH), a 39-kDa iron-sulfur- and FAD-containing reductase (MMOR), and a 16-kDa regulatory protein (MMOB). The thermodynamic stability and kinetics of formation of complexes between oxidized MMOH and MMOB or MMOR were measured by isothermal titration calorimetry and stopped-flow fluorescence spectroscopy at temperatures ranging from 3.3 to 45 degrees C. The results, in conjunction with data from equilibrium analytical ultracentrifugation studies of MMOR and MMOB, indicate that free MMOR and MMOB exist as monomers in solution and bind MMOH with 2:1 stoichiometry. The role of component interactions in the catalytic mechanism of sMMO was investigated through simultaneous measurement of oxidase and hydroxylase activities as a function of varied protein component concentrations during steady-state turnover. The partitioning of oxidase and hydroxylase activities of sMMO is highly dependent on both the MMOR concentration and the nature of the organic substrate. In particular, NADH oxidation is significantly uncoupled from methane hydroxylation at MMOR concentrations exceeding 20% of the hydroxylase concentration but remains tightly coupled to propylene epoxidation at MMOR concentrations ranging up to the MMOH concentration. The steady-state kinetic data were fit to numerical simulations of models that include both the oxidase activities of free MMOR and of MMOH/MMOR complexes and the hydroxylase activity of MMOH/MMOB complexes. The data were well described by a model in which MMOR and MMOB bind noncompetitively at distinct interacting sites on the hydroxylase. MMOB manifests its regulatory effects by differentially accelerating intermolecular electron transfer from MMOR to MMOH containing bound substrate and product in a manner consistent with its activating and inhibitory effects on the hydroxylase. PMID:10504247

  12. Electron-transfer reactions of the reductase component of soluble methane monooxygenase from Methylococcus capsulatus (Bath).

    PubMed

    Kopp, D A; Gassner, G T; Blazyk, J L; Lippard, S J

    2001-12-11

    Soluble methane monooxygenase (sMMO) catalyzes the hydroxylation of methane by dioxygen to afford methanol and water, the first step of carbon assimilation in methanotrophic bacteria. This enzyme comprises three protein components: a hydroxylase (MMOH) that contains a dinuclear nonheme iron active site; a reductase (MMOR) that facilitates electron transfer from NADH to the diiron site of MMOH; and a coupling protein (MMOB). MMOR uses a noncovalently bound FAD cofactor and a [2Fe-2S] cluster to mediate electron transfer. The gene encoding MMOR was cloned from Methylococcus capsulatus (Bath) and expressed in Escherichia coli in high yield. Purified recombinant MMOR was indistinguishable from the native protein in all aspects examined, including activity, mass, cofactor content, and EPR spectrum of the [2Fe-2S] cluster. Redox potentials for the FAD and [2Fe-2S] cofactors, determined by reductive titrations in the presence of indicator dyes, are FAD(ox/sq), -176 +/- 7 mV; FAD(sq/hq), -266 +/- 15 mV; and [2Fe-2S](ox/red), -209 +/- 14 mV. The midpoint potentials of MMOR are not altered by the addition of MMOH, MMOB, or both MMOH and MMOB. The reaction of MMOR with NADH was investigated by stopped-flow UV-visible spectroscopy, and the kinetic and spectral properties of intermediates are described. The effects of pH on the redox properties of MMOR are described and exploited in pH jump kinetic studies to measure the rate constant of 130 +/- 17 s(-)(1) for electron transfer between the FAD and [2Fe-2S] cofactors in two-electron-reduced MMOR. The thermodynamic and kinetic parameters determined significantly extend our understanding of the sMMO system. PMID:11732913

  13. Inactivation of the particulate methane monooxygenase (pMMO) in Methylococcus capsulatus (Bath) by acetylene.

    PubMed

    Pham, Minh D; Lin, Ya-Ping; Van Vuong, Quan; Nagababu, Penumaka; Chang, Brian T-A; Ng, Kok Yaoh; Chen, Chein-Hung; Han, Chau-Chung; Chen, Chung-Hsuan; Li, Mai Suan; Yu, Steve S-F; Chan, Sunney I

    2015-12-01

    Acetylene (HCCH) has a long history as a mechanism-based enzyme inhibitor and is considered an active-site probe of the particulate methane monooxygenase (pMMO). Here, we report how HCCH inactivates pMMO in Methylococcus capsulatus (Bath) by using high-resolution mass spectrometry and computational simulation. High-resolution MALDI-TOF MS of intact pMMO complexes has allowed us to confirm that the enzyme oxidizes HCCH to the ketene (C2H2O) intermediate, which then forms an acetylation adduct with the transmembrane PmoC subunit. LC-MS/MS analysis of the peptides derived from in-gel proteolytic digestion of the protein subunit identifies K196 of PmoC as the site of acetylation. No evidence is obtained for chemical modification of the PmoA or PmoB subunit. The inactivation of pMMO by a single adduct in the transmembrane PmoC domain is intriguing given the complexity of the structural fold of this large membrane-protein complex as well as the complicated roles played by the various metal cofactors in the enzyme catalysis. Computational studies suggest that the entry of hydrophobic substrates to, and migration of products from, the catalytic site of pMMO are controlled tightly within the transmembrane domain. Support of these conclusions is provided by parallel experiments with two related alkynes: propyne (CH3CCH) and trifluoropropyne (CF3CCH). Finally, we discuss the implication of these findings to the location of the catalytic site in pMMO. PMID:26275807

  14. Expression, purification and biochemical characterization of a family 6 carboxylesterase from Methylococcus capsulatus (bath).

    PubMed

    Soni, Surabhi; Odaneth, Annamma A; Lali, Arvind M; Chandrayan, Sanjeev K

    2016-06-01

    The genome of Methylococcus capsulatus (bath) encodes a protein R-est6 that is annotated as a lipase family 3 protein. The phylogenetic and the sequence analyses linked this protein to the family 6 carboxylesterase. The gene encoding R-est6 was cloned and overexpressed in Escherichia coli and the recombinant 6x-His tagged protein was purified by Ni-NTA affinity chromatography. The buffers used in the purification were modified by adding 1% glycerol instead of the salt to prevent the protein aggregation. Far UV-CD spectrum and gel filtration chromatography of the purified R-est6 confirmed that the protein was well folded like a typical α/β hydrolase and had the quaternary structure of a tetramer, in addition to a compact monomer. The optimum pH was in the range of 7.0-9.0 and the optimum temperature was at 55 °C for the hydrolysis of pNP-butyrate. As expected, being a member of the family 6 carboxylesterase, R-est6 hydrolyzed triglycerides, pNP esters of the small and the medium fatty acid chain esters and an aryl ester-phenyl acetate. However, R-est6 was also found to hydrolyze the long-chain fatty acid ester which had never been reported for the family 6 carboxylesterase. Additionally, R-est6 was stable and active in the different water-miscible organic solvents. Therefore, the broad substrate range and the structural stability of R-est6 would be advantageous for its application in industrial processes. PMID:26899525

  15. Squalene-hopene cyclase from Methylococcus capsulatus (Bath): a bacterium producing hopanoids and steroids.

    PubMed

    Tippelt, A; Jahnke, L; Poralla, K

    1998-03-30

    We report the cloning and characterisation of the Methylococcus capsulatus shc gene, which encodes the squalene-hopene cyclase (SHC). This enzyme catalyses the complex cyclization of squalene to the pentacyclic triterpene skeleton of hopanoids and represents the key reaction in this biosynthesis. Using a combination of PCR amplification and DNA hybridization, two overlapping 2.6 kb PstI and 3.3 kb SalI DNA fragments were cloned bearing a 1962 bp open reading frame encoding a 74 kDa protein with 654 amino acids and a predicted isoelectric point at about pH 6.3. The deduced amino acid sequence of the M. capsulatus shc gene showed significant similarity to known prokaryotic SHCs and to a lesser degree to the related eukaryotic oxidosqualene cyclases (OSCs). Like other triterpene cyclases, the M. capsulatus SHC contains seven so-called QW-motifs as well as an aspartate-rich domain. The recombinant M. capsulatus SHC was expressed in Escherichia coli and in vitro activity of the recombinant cyclase was demonstrated using crude cell-free lysate or solubilized membrane preparation. The cyclization products hop-22-ene and hopan-22-ol (diplopterol) were identified by GC and GC-MS. PMID:9555026

  16. Structure and protein–protein interactions of methanol dehydrogenase from Methylococcus capsulatus (Bath)

    SciTech Connect

    Culpepper, Megen A.; Rosenzweig, Amy C.

    2014-10-07

    In the initial steps of their metabolic pathway, methanotrophic bacteria oxidize methane to methanol with methane monooxygenases (MMOs) and methanol to formaldehyde with methanol dehydrogenases (MDHs). Several lines of evidence suggest that the membrane-bound or particulate MMO (pMMO) and MDH interact to form a metabolic supercomplex. To further investigate the possible existence of such a supercomplex, native MDH from Methylococcus capsulatus (Bath) has been purified and characterized by size exclusion chromatography with multi-angle light scattering and X-ray crystallography. M. capsulatus (Bath) MDH is primarily a dimer in solution, although an oligomeric species with a molecular mass of ~450–560 kDa forms at higher protein concentrations. The 2.57 Å resolution crystal structure reveals an overall fold and α₂β₂ dimeric architecture similar to those of other MDH structures. In addition, biolayer interferometry studies demonstrate specific protein–protein interactions between MDH and M. capsulatus (Bath) pMMO as well as between MDH and the truncated recombinant periplasmic domains of M. capsulatus (Bath) pMMO (spmoB). These interactions exhibit KD values of 833 ± 409 nM and 9.0 ± 7.7 μM, respectively. The biochemical data combined with analysis of the crystal lattice interactions observed in the MDH structure suggest a model in which MDH and pMMO associate not as a discrete, stoichiometric complex but as a larger assembly scaffolded by the intracytoplasmic membranes.

  17. Multiple polypeptide forms observed in two-dimensional gels of Methylococcus capsulatus (Bath) polypeptides are generated during the separation procedure.

    PubMed

    Berven, Frode S; Karlsen, Odd A; Murrell, J Colin; Jensen, Harald B

    2003-02-01

    We have examined two-dimensional electrophoresis (2-DE) gel maps of polypeptides from the Gram-negative bacterium Methylococcus capsulatus (Bath) and found the same widespread trains of spots as often reported in 2-DE gels of polypeptides of other Gram-negative bacteria. Some of the trains of polypeptides, both from the outer membrane and soluble protein fraction, were shown to be generated during the separation procedure of 2-DE, and not by covalent post-translational modifications. The trains were found to be regenerated when rerunning individual polypeptide spots. The polypeptides analysed giving this type of trains were all found to be classified as stable polypeptides according to the instability index of Guruprasad et al. (Protein Eng. 1990, 4, 155-161). The phenomenon most likely reflects conformational equilibria of polypeptides arising from the experimental conditions used, and is a clear drawback of the standard 2-DE procedure, making the gel picture unnecessarily complex to analyse. PMID:12601748

  18. The effects of growth temperature on the methyl sterol and phospholipid fatty acid composition of Methylococcus capsulatus (Bath)

    NASA Technical Reports Server (NTRS)

    Jahnke, Linda L.

    1992-01-01

    Growth of Methylococcus capsulatus (Bath) at temperatures ranging from 30 to 50 C resulted in changes to the whole cell lipid constituents. As temperature was lowered, the overall proportion of hexadecenoic acid (C16:1) increased, and the relative proportions of the Delta9, Delta10, and Delta11 C16:1 double bond positional isomers changed. Methyl sterol content also increased as the growth temperature was lowered. The highest amounts of methyl sterol were found in 30 C cells and the lowest in 50 C cells (sterol-phospholipid ratios of 0.077 and 0.013, respectively). The data are consistent with a membrane modulating role for the sterol produced by this prokaryotic organism.

  19. The effects of growth temperature on the methyl sterol and phospholipid fatty acid composition of Methylococcus capsulatus (Bath)

    NASA Technical Reports Server (NTRS)

    Jahnke, L. L.

    1992-01-01

    Growth of Methylococcus capsulatus (Bath) at temperatures ranging from 30 to 50 degrees C resulted in changes to the whole cell lipid constituents. As temperature was lowered, the overall proportion of hexadecenoic acid (C16:1) increased, and the relative proportions of the delta 9, delta 10 and delta 11 C16:1 double bond positional isomers changed. Methyl sterol content also increased as the growth temperature was lowered. The highest amounts of methyl sterol were found in 30 degrees C cells and the lowest in 50 degrees C cells (sterol-phospholipid ratios of 0.077 and 0.013, respectively). The data are consistent with a membrane modulating role for the sterol produced by this prokaryotic organism.

  20. Characterization of the recombinant pyrophosphate-dependent 6-phosphofructokinases from Methylomicrobium alcaliphilum 20Z and Methylococcus capsulatus Bath.

    PubMed

    Khmelenina, Valentina N; Rozova, Olga N; Trotsenko, Yuri A

    2011-01-01

    The Embden-Meyerhof-Parnas (EMP) glycolysis is the starting point of the core carbon metabolism. Aerobic methanotrophs possessing activity of the pyrophosphate-dependent 6-phosphofructokinase (PPi-PFK) instead of the classical glycolytic enzyme ATP-dependent 6-phosphofructokinase (ATP-PFK) are promising model bacteria for elucidation of the role of inorganic pyrophosphate (PPi) and PPi-dependent glycolysis in microorganisms. Characterization of the His(6)-tagged PPi-PFKs from two methanotrophs, halotolerant alkaliphilic Methylomicrobium alcaliphilum 20Z and thermotolerant Methylococcus capsulatus Bath, showed differential capabilities of PPi-PFKs to phosphorylate sedoheptulose-7-phosphate and this property correlated well with the metabolic patterns of these bacteria assimilating C(1) substrate either via the ribulosemonophosphate (RuMP) pathway (Mm. alcaliphilum 20Z) or simultaneously via the RuMP and serine pathways and the Calvin cycle (Mc. capsulatus Bath). Analysis of the genomic draft of Mm. alcaliphilum 20Z (https://www.genoscope.cns.fr/agc/mage) has provided in silico evidence for the existence of a PPi-dependent pyruvate-phosphate dikinase (PPDK). Expression of the ppdk gene at oxygen limitation along with the presence of PPi-PFK in Mm. alcaliphilum 20Z implied functioning of PPi-dependent glycolysis and PPi recycling under conditions when oxidative phosphorylation is hampered. PMID:21419911

  1. The Surface-Associated and Secreted MopE Protein of Methylococcus capsulatus (Bath) Responds to Changes in the Concentration of Copper in the Growth Medium

    PubMed Central

    Karlsen, Odd A.; Berven, Frode S.; Stafford, Graham P.; Larsen, Øivind; Murrell, J. Colin; Jensen, Harald B.; Fjellbirkeland, Anne

    2003-01-01

    Expression of surface-associated and secreted protein MopE of the methanotrophic bacterium Methylococcus capsulatus (Bath) in response to the concentration of copper ions in the growth medium was investigated. The level of protein associated with the cells and secreted to the medium changed when the copper concentration in the medium varied and was highest in cells exposed to copper stress. PMID:12676726

  2. The surface-associated and secreted MopE protein of Methylococcus capsulatus (Bath) responds to changes in the concentration of copper in the growth medium.

    PubMed

    Karlsen, Odd A; Berven, Frode S; Stafford, Graham P; Larsen, Øivind; Murrell, J Colin; Jensen, Harald B; Fjellbirkeland, Anne

    2003-04-01

    Expression of surface-associated and secreted protein MopE of the methanotrophic bacterium Methylococcus capsulatus (Bath) in response to the concentration of copper ions in the growth medium was investigated. The level of protein associated with the cells and secreted to the medium changed when the copper concentration in the medium varied and was highest in cells exposed to copper stress. PMID:12676726

  3. Role of multiple gene copies in particulate methane monooxygenase activity in the methane-oxidizing bacterium Methylococcus capsulatus Bath.

    PubMed

    Stolyar, S; Costello, A M; Peeples, T L; Lidstrom, M E

    1999-05-01

    Genes for the subunits of particulate methane monooxygenase, PmoABC, have been sequenced from the gamma-proteobacterial methanotroph Methylococcus capsulatus Bath. M. capsulatus Bath contains two complete copies of pmoCAB, as well as a third copy of pmoC. The two pmoCAB regions were almost identical at the nucleotide sequence level, differing in only 13 positions in 3183 bp. At the amino acid level, each translated gene product contained only one differing residue in each copy. However, the pmoC3 sequence was more divergent from the two other pmoC copies at both the far N-terminus and far C-terminus. Chromosomal insertion mutations were generated in all seven genes. Null mutants could not be obtained for pmoC3, suggesting that it may play an essential role in growth on methane. Null mutants were obtained for pmoC1, pmoC2, pmoA1, pmoA2, pmoB1 and pmoB2. All of these mutants grew on methane, demonstrating that both gene copies were functional. Copy 1 mutants showed about two-thirds of the wild-type whole-cell methane oxidation rate, while copy 2 mutants showed only about one-third of the wild-type rate, indicating that both gene copies were necessary for wild-type particulate methane monooxygenase activity. It was not possible to obtain double null mutants that were defective in both pmo copies, which may indicate that some expression of pMMO is important for growth. PMID:10376840

  4. The membrane-associated form of methane mono-oxygenase from Methylococcus capsulatus (Bath) is a copper/iron protein.

    PubMed Central

    Basu, Piku; Katterle, Bettina; Andersson, K Kristoffer; Dalton, Howard

    2003-01-01

    A protocol has been developed which permits the purification of a membrane-associated methane-oxidizing complex from Methylococcus capsulatus (Bath). This complex has approximately 5 fold higher specific activity than any purified particulate methane mono-oxygenase (pMMO) previously reported from M. capsulatus (Bath). This efficiently functioning methane-oxidizing complex consists of the pMMO hydroxylase (pMMOH) and an unidentified component we have assigned as a potential pMMO reductase (pMMOR). The complex was isolated by solubilizing intracytoplasmic membrane preparations containing the high yields of active membrane-bound pMMO (pMMO(m)), using the non-ionic detergent dodecyl-beta-D-maltoside, to yield solubilized enzyme (pMMO(s)). Further purification gave rise to an active complex (pMMO(c)) that could be resolved (at low levels) by ion-exchange chromatography into two components, the pMMOH (47, 27 and 24 kDa subunits) and the pMMOR (63 and 8 kDa subunits). The purified complex contains two copper atoms and one non-haem iron atom/mol of enzyme. EPR spectra of preparations grown with (63)Cu indicated that the copper ion interacted with three or four nitrogenic ligands. These EPR data, in conjunction with other experimental results, including the oxidation by ferricyanide, EDTA treatment to remove copper and re-addition of copper to the depleted protein, verified the essential role of copper in enzyme catalysis and indicated the implausibility of copper existing as a trinuclear cluster. The EPR measurements also demonstrated the presence of a tightly bound mononuclear Fe(3+) ion in an octahedral environment that may well be exchange-coupled to another paramagnetic species. PMID:12379148

  5. The membrane-associated form of methane mono-oxygenase from Methylococcus capsulatus (Bath) is a copper/iron protein.

    PubMed

    Basu, Piku; Katterle, Bettina; Andersson, K Kristoffer; Dalton, Howard

    2003-01-15

    A protocol has been developed which permits the purification of a membrane-associated methane-oxidizing complex from Methylococcus capsulatus (Bath). This complex has approximately 5 fold higher specific activity than any purified particulate methane mono-oxygenase (pMMO) previously reported from M. capsulatus (Bath). This efficiently functioning methane-oxidizing complex consists of the pMMO hydroxylase (pMMOH) and an unidentified component we have assigned as a potential pMMO reductase (pMMOR). The complex was isolated by solubilizing intracytoplasmic membrane preparations containing the high yields of active membrane-bound pMMO (pMMO(m)), using the non-ionic detergent dodecyl-beta-D-maltoside, to yield solubilized enzyme (pMMO(s)). Further purification gave rise to an active complex (pMMO(c)) that could be resolved (at low levels) by ion-exchange chromatography into two components, the pMMOH (47, 27 and 24 kDa subunits) and the pMMOR (63 and 8 kDa subunits). The purified complex contains two copper atoms and one non-haem iron atom/mol of enzyme. EPR spectra of preparations grown with (63)Cu indicated that the copper ion interacted with three or four nitrogenic ligands. These EPR data, in conjunction with other experimental results, including the oxidation by ferricyanide, EDTA treatment to remove copper and re-addition of copper to the depleted protein, verified the essential role of copper in enzyme catalysis and indicated the implausibility of copper existing as a trinuclear cluster. The EPR measurements also demonstrated the presence of a tightly bound mononuclear Fe(3+) ion in an octahedral environment that may well be exchange-coupled to another paramagnetic species. PMID:12379148

  6. Why OrfY? Characterization of MMOD, a long overlooked component of the soluble methane monooxygenase from Methylococcus capsulatus (Bath).

    PubMed

    Merkx, Maarten; Lippard, Stephen J

    2002-02-22

    Soluble methane monooxygenase (sMMO) has been studied intensively to understand the mechanism by which it catalyzes the remarkable oxidation of methane to methanol. The cluster of genes that encode for the three characterized protein components of sMMO (MMOH, MMOB, and MMOR) contains an additional open reading frame (orfY) of unknown function. In the present study, MMOD, the protein encoded by orfY, was overexpressed as a fusion protein in Escherichia coli. Pure MMOD was obtained in high yields after proteolytic cleavage and a two-step purification procedure. Western blot analysis of Methylococcus capsulatus (Bath) soluble cell extracts showed that MMOD is expressed in the native organism although at significantly lower levels than the other sMMO proteins. The cofactorless MMOD protein is a potent inhibitor of sMMO activity and binds to the hydroxylase protein (MMOH) with an affinity similar to that of MMOB and MMOR. The addition of up to 2 MMOD per MMOH results in changes in the optical spectrum of the hydroxylase that suggest the formation of a (micro-oxo)diiron(III) center in a fraction of the MMOH-MMOD complexes. Possible functions for MMOD are discussed, including a role in the assembly of the MMOH diiron center similar to that suggested for DmpK, a protein that shares some properties with MMOD. PMID:11709550

  7. Three-dimensional structure determination of a protein supercomplex that oxidizes methane to formaldehyde in Methylococcus capsulatus (Bath).

    PubMed

    Myronova, Natalia; Kitmitto, Ashraf; Collins, Richard F; Miyaji, Aki; Dalton, Howard

    2006-10-01

    The oxidation of methane to methanol in methanotrophs is catalyzed by the enzyme methane monooxygenase (MMO). Two distinct forms of this enzyme exist, a soluble cytoplasmic MMO (sMMO) and a membrane-bound particulate form (pMMO). The active protein complex termed pMMO-C was purified recently from Methylococcus capsulatus (Bath). The complex consists of pMMO hydroxylase and an additional component pMMO-R, which was proposed to be the reductase for the pMMO complex. Further study of this complex has led here to the proposal that the pMMO-R is in fact methanol dehydrogenase, the subsequent enzyme in the methane oxidation pathway by methanotrophs. We describe here the biochemical and biophysical characterization of a stable purified complex of pMMO hydroxylase (pMMO-H) with methanol dehydrogenase (MDH) and report the first three-dimensional (3D) structure, determined by cryoelectron microscopy and single particle analysis to approximately 16 A resolution. The 3D structure reported here provides the first insights into the supramolecular organization of pMMO with MDH. These studies of pMMO-MDH complexes have provided further understanding of the structural basis for the particular functions of the enzymes in this system which might also be of relevance to the complete process of methane oxidation by methanotrophs under high copper concentration in the environment. PMID:17002291

  8. Kinetic and spectroscopic characterization of intermediates and component interactions in reactions of methane monooxygenase from methylococcus capsulatus (Bath)

    SciTech Connect

    Liu, K.E.; Valentine, A.M.; Salifoglou, A.; Lippard, S.J.; Wang, D.; Huynh, B.H.; Edmondson, D.E.

    1995-10-18

    We describe mechanistic studies of the soluble methane monooxygenase (sMMO) enzyme system from Methylococcus capsulatus (Bath). Interactions among the three sMMO components, the hydroxylase (H), reductase (R), and protein B (B), were investigated by monitoring conversion of nitrobenzene to nitrophenol under both single turnover and catalytic conditions. During catalytic turnover, hydroxylation occurs to afford 3-nitrophenol (43%) and 4-nitrophenol (57%), whereas hydroxylation takes place exclusively (> 95%) to give 4-nitrophenol under single turnover conditions in the absence of reductase. Protein B exerts a strong influence on single turnover reactions of nitrobenzene, with optimal rate constants and yields obtained by using 1.5-2 equiv of protein R per equivalent of hydroxylase. The temperature dependence of these kinetic values was determined. Changes in dioxygen concentration and pH, as well as exchange of solvent accessible protons with D{sub 2}O, did not significantly affect the rate constants for either of these processes, the implications of which for the kinetic mechanism are discussed. From the present and related evidence, structures for H{sub peroxo} and Q are proposed. 54 refs., 11 figs., 4 tabs.

  9. Tritiated chiral alkanes as substrates for soluble methane monooxygenase from Methylococcus capsulatus (Bath): Probes for the mechanism of hydroxylation

    SciTech Connect

    Valentine, A.M.; Liu, K.E.; Komar-Panicucci, S.; Lippard, S.J.; Wilkinson, B.; Priestley, N.D.; Floss, H.G.; Williams, P.G.; Morimoto, Hiromi

    1997-02-26

    The tritiated chiral alkanes (S)-[1-{sup 2}H{sub 1}, 1-{sup 3}H]ethane, (R)-[1-{sup 2}H{sub 1},1-{sup 3}H]ethane, (S)-[1-{sup 2}H{sub 1},1-{sup 3}H]butane, (R)-[1-{sup 2}H{sub 1}, 1-{sup 3}H]butane, (S)-[2-{sup 3}H]butane, (R)-[2-{sup 3}H]butane, and racemic [2-{sup 3}H]butane were oxidized by soluble methane monooxygenase (sMMO) from Methylococcus capsulatus (Bath), and the absolute stereochemistry of the resulting product alcohols was determined in order to probe the mechanism of substrate hydroxylation. When the hydroxylations were performed with purified hydroxylase but only a partially purified cellular extract for the coupling and reductase proteins, different product distributions were observed. These apparently anomalous results could be explained by invoking exchange of hydrogen atoms at the {alpha} carbon of the product alcohols. The characteristics of this exchange reaction are discussed. Together with the mechanistic information available from a range of substrate probes, the results are best accounted for by a nonsynchronous concerted process involving attack on the C-H bond by one or more of several pathways discussed in the text. 65 refs., 5 figs., 3 tabs.

  10. Primary structure of cytochrome c' of Methylococcus capsulatus Bath: evidence of a phylogenetic link between P460 and c'-type cytochromes.

    PubMed

    Bergmann, D J; Zahn, J A; DiSpirito, A A

    2000-01-01

    Cytochrome c' of Methylococcus capsulatus Bath is involved in electron flow from the enzyme responsible for hydroxylamine oxidation, cytochrome P460, to cytochrome C555. This cytochrome is spectrally similar to other cytochromes c' but is larger (16,000 Da) and has a lower midpoint potential (-205 mV). By a combination of Edman degradation, mass spectroscopy, and gene sequencing, we have obtained the primary structure of cytochrome c' from M. capsulatus Bath. The cytochrome shows low sequence similarity to other cytochromes c', only residues R12, Y53, G56, and the C-terminal heme-binding region (GXXCXXCHXXXK) being conserved. In contrast, cytochrome c' from M. capsulatus Bath shows considerable sequence similarity to cytochromes P460 from M. capsulatus Bath (31% identity) and from Nitrosomonas europaea (18% identity). This suggests that P460-type cytochromes may have originated from a c'-type cytochrome which developed a covalent cross-link between a lysine residue and the c'-heme. PMID:10648101

  11. The noncommensal bacterium Methylococcus capsulatus (Bath) ameliorates dextran sulfate (Sodium Salt)-Induced Ulcerative Colitis by influencing mechanisms essential for maintenance of the colonic barrier function.

    PubMed

    Kleiveland, Charlotte R; Hult, Lene T Olsen; Spetalen, Signe; Kaldhusdal, Magne; Christofferesen, Trine Eker; Bengtsson, Oskar; Romarheim, Odd Helge; Jacobsen, Morten; Lea, Tor

    2013-01-01

    Dietary inclusion of a bacterial meal has recently been shown to efficiently abolish soybean meal-induced enteritis in Atlantic salmon. The objective of this study was to investigate whether inclusion of this bacterial meal in the diet could abrogate disease development in a murine model of epithelial injury and colitis and thus possibly have therapeutic potential in human inflammatory bowel disease. C57BL/6N mice were fed ad libitum a control diet or an experimental diet containing 254 g/kg of body weight BioProtein, a bacterial meal consisting of Methylococcus capsulatus (Bath), together with the heterogenic bacteria Ralstonia sp., Brevibacillus agri, and Aneurinibacillus sp. At day 8, colitis was induced by 3.5% dextran sulfate sodium (DSS) ad libitum in the drinking water for 6 days. Symptoms of DSS treatment were less profound after prophylactic treatment with the diet containing the BioProtein. Colitis-associated parameters such as reduced body weight, colon shortening, and epithelial damage also showed significant improvement. Levels of acute-phase reactants, proteins whose plasma concentrations increase in response to inflammation, and neutrophil infiltration were reduced. On the other, increased epithelial cell proliferation and enhanced mucin 2 (Muc2) transcription indicated improved integrity of the colonic epithelial layer. BioProtein mainly consists of Methylococcus capsulatus (Bath) (88%). The results that we obtained when using a bacterial meal consisting of M. capsulatus (Bath) were similar to those obtained when using BioProtein in the DSS model. Our results show that a bacterial meal of the noncommensal bacterium M. capsulatus (Bath) has the potential to attenuate DSS-induced colitis in mice by enhancing colonic barrier function, as judged by increased epithelial proliferation and increased Muc2 transcription. PMID:23064342

  12. The Noncommensal Bacterium Methylococcus capsulatus (Bath) Ameliorates Dextran Sulfate (Sodium Salt)-Induced Ulcerative Colitis by Influencing Mechanisms Essential for Maintenance of the Colonic Barrier Function

    PubMed Central

    Hult, Lene T. Olsen; Spetalen, Signe; Kaldhusdal, Magne; Christofferesen, Trine Eker; Bengtsson, Oskar; Romarheim, Odd Helge; Jacobsen, Morten; Lea, Tor

    2013-01-01

    Dietary inclusion of a bacterial meal has recently been shown to efficiently abolish soybean meal-induced enteritis in Atlantic salmon. The objective of this study was to investigate whether inclusion of this bacterial meal in the diet could abrogate disease development in a murine model of epithelial injury and colitis and thus possibly have therapeutic potential in human inflammatory bowel disease. C57BL/6N mice were fed ad libitum a control diet or an experimental diet containing 254 g/kg of body weight BioProtein, a bacterial meal consisting of Methylococcus capsulatus (Bath), together with the heterogenic bacteria Ralstonia sp., Brevibacillus agri, and Aneurinibacillus sp. At day 8, colitis was induced by 3.5% dextran sulfate sodium (DSS) ad libitum in the drinking water for 6 days. Symptoms of DSS treatment were less profound after prophylactic treatment with the diet containing the BioProtein. Colitis-associated parameters such as reduced body weight, colon shortening, and epithelial damage also showed significant improvement. Levels of acute-phase reactants, proteins whose plasma concentrations increase in response to inflammation, and neutrophil infiltration were reduced. On the other, increased epithelial cell proliferation and enhanced mucin 2 (Muc2) transcription indicated improved integrity of the colonic epithelial layer. BioProtein mainly consists of Methylococcus capsulatus (Bath) (88%). The results that we obtained when using a bacterial meal consisting of M. capsulatus (Bath) were similar to those obtained when using BioProtein in the DSS model. Our results show that a bacterial meal of the noncommensal bacterium M. capsulatus (Bath) has the potential to attenuate DSS-induced colitis in mice by enhancing colonic barrier function, as judged by increased epithelial proliferation and increased Muc2 transcription. PMID:23064342

  13. Temperature Affects Fatty Acids In Methylococcus Capsulatus

    NASA Technical Reports Server (NTRS)

    Jahnke, Linda L.

    1993-01-01

    According to report, temperature of growth of thermotolerant, methane-oxidizing bacterium Methylococcus capsulatus (Bath) affects both proportion of monounsaturated fatty acids and cis/trans ratio of these acids in cell membrane. Because suboptimum growth temperature is potential stress factor, it may be possible to use such cis/trans ratios as indices of stresses upon methane-oxidizing microbial communities. Research in microbiology of methanotrophs increasing because of possible commercial exploitation of these organisms as biocatalysts or as sources of useful polymers; knowledge of effect of temperature on ability of methanotrophs to utilize methane useful in optimization of conditions of growth.

  14. Crystal structures of the methane monooxygenase hydroxylase from Methylococcus capsulatus (Bath): implications for substrate gating and component interactions.

    PubMed

    Rosenzweig, A C; Brandstetter, H; Whittington, D A; Nordlund, P; Lippard, S J; Frederick, C A

    1997-10-01

    The crystal structure of the nonheme iron-containing hydroxylase component of methane monooxygenase hydroxylase (MMOH) from Methylococcus capsulatus (Bath) has been solved in two crystal forms, one of which was refined to 1.7 A resolution. The enzyme is composed of two copies each of three subunits (alpha 2 beta 2 gamma 2), and all three subunits are almost completely alpha-helical, with the exception of two beta hairpin structures in the alpha subunit. The active site of each alpha subunit contains one dinuclear iron center, housed in a four-helix bundle. The two iron atoms are octahedrally coordinated by 2 histidine and 4 glutamic acid residues as well as by a bridging hydroxide ion, a terminal water molecule, and at 4 degrees C, a bridging acetate ion, which is replaced at -160 degrees C with a bridging water molecule. Comparison of the results for two crystal forms demonstrates overall conservation and relative orientation of the domain structures. The most prominent structural differences identified between the two crystal forms is in an altered side chain conformation for Leu 110 at the active site cavity. We suggest that this residue serves as one component of a hydrophobic gate controlling access of substrates to and products from the active site. The leucine gate may be responsible for the effect of the B protein component on the reactivity of the reduced hydroxylase with dioxygen. A potential reductase binding site has been assigned based on an analysis of crystal packing in the two forms and corroborated by inhibition studies with a synthetic peptide corresponding to the proposed docking position. PMID:9329079

  15. Structural characterization by EXAFS spectroscopy of the binuclear iron center in protein A of methane monooxygenase from methylococcus capsulatus (bath)

    SciTech Connect

    Ericson, A.; Hedman, B.; Hodgson, K.O.; Green, J.; Dalton, H.; Bentsen, J.G.; Beer, R.H.; Lippard, S.J.

    1988-03-30

    Soluble methane monooxygenase (MMO) from Methylococcus capsulatus (Bath) activates dioxygen for incorporation into a remarkable variety of substrates including methane, which is required for bacterial growth (eq 1). MMO is a three component CH/sub 4/ + NADH + H/sup +/ + O/sub 2/ /sup MMO/ ..-->.. CH/sub 3/OH + NAD/sup +/ + H/sub 2/O (1) enzyme. Protein A (M/sub r/ 210,000), believed to be the oxygenase component, contains two iron atoms. Protein B (M/sub r/ 15,700) serves a regulatory function and lacks prosthetic groups, while protein C, the reductase component of the enzyme, is an iron-sulfur flavoprotein (M/sub r/ 42,000) responsible for electron transfer from NADH to protein A. Recently, a binuclear iron center was postulated to occur in protein A based on the finding that one-electron reduction gives rise to electron spin resonance (ESR) signals (g 1.95, 1.88, 1.78) very similar to those observed for the binuclear mixed-valence Fe/sub 2/(III,II) centers in semimet hemerythrin (Hr) and purple acid phosphatase (PAP). In conjunction with model studies, extended X-ray absorption fine structure (EXAFS) spectroscopy has proven to be a powerful method for identifying bridged binuclear iron centers in Hr, ribonucleotide reductase (RR), and PAP. Here the authors report iron K-edge EXAFS results on semireduced protein A of MMO which support the occurrence of a binuclear iron center (Fe-Fe distance, 3.41 A), with no short ..mu..-oxo bridge.

  16. Insights into the obligate methanotroph Methylococcus capsulatus.

    PubMed

    Kelly, Donovan P; Anthony, Christopher; Murrell, J Colin

    2005-05-01

    Completion of the genome sequence of Methylococcus capsulatus Bath is an important event in molecular microbiology, and an achievement for which the authors deserve congratulation. M. capsulatus, along with other methanotrophs, has been the subject of intense biochemical and molecular study because of its role in the global carbon cycle: the conversion of biogenic methane to carbon dioxide. The methane monooxygenase enzymes that are central to this process also have high biotechnological potential. Analysis of the genome sequence will potentially accelerate elucidation of the regulation of methane-dependent metabolism in obligate methanotrophs, and help explain the cause of obligate methanotrophy, the phenomenon making most methanotrophs unable to grow on any substrates other than methane and a very small number of other one-carbon compounds. PMID:15866035

  17. Determination of the carbon kinetic isotope effects on propane hydroxylation mediated by the methane monooxygenases from Methylococcus capsulatus (Bath) by using stable carbon isotopic analysis.

    PubMed

    Huang, Ded-Shih; Wu, Suh-Huey; Wang, Yane-Shih; Yu, Steve S-F; Chan, Sunney I

    2002-08-01

    Authentic propane with known position-specific carbon isotope composition at each carbon atom was subjected to hydroxylation by the particulate and soluble methane monooxygenase (pMMO and sMMO) from Methylococcus capsulatus (Bath), and the corresponding position-specific carbon isotope content was redetermined for the product 2-propanol. Neither the reaction mediated by pMMO nor that with sMMO showed an intermolecular (12)C/(13)C kinetic isotope effect effect on the propane hydroxylation at the secondary carbon; this indicates that there is little structural change at the carbon center attacked during formation of the transition state in the rate-determining step. This finding is in line with the concerted mechanism proposed for pMMO (Bath), and suggested for sMMO (Bath), namely, direct side-on insertion of an active "O" species across the C-H bond, as has been previously reported for singlet carbene insertion. PMID:12203974

  18. Physiological evidence for the presence of a cis-trans isomerase of unsaturated fatty acids in Methylococcus capsulatus Bath to adapt to the presence of toxic organic compounds.

    PubMed

    Löffler, Claudia; Eberlein, Christian; Mäusezahl, Ines; Kappelmeyer, Uwe; Heipieper, Hermann J

    2010-07-01

    The physiology of the response in the methanotrophic bacterium Methylococcus capsulatus Bath towards thermal and solvent stress was studied. A systematic investigation of the toxic effects of organic compounds (chlorinated phenols and alkanols) on the growth of this bacterium was carried out. The sensitivity to the tested alkanols correlated with their chain length and hydrophobicity; methanol was shown to be an exception to which the cells showed a very high tolerance. This can be explained by the adaptation of these bacteria to growth on C1 compounds. On the other hand, M. capsulatus Bath was very sensitive towards the tested chlorinated phenols. The high toxic effect of phenolic compounds on methanotrophic bacteria might be explained by the occurrence of toxic reactive oxygen species. In addition, a physiological proof of the presence of cis-trans isomerization as a membrane-adaptive response mechanism in M. capsulatus was provided. This is the first report on physiological evidence for the presence of the unique postsynthetic membrane-adaptive response mechanism of the cis-trans isomerization of unsaturated fatty acids in a bacterium that does not belong to the genera Pseudomonas and Vibrio where this mechanism was already reported and described extensively. PMID:20487020

  19. Comparison of EPR-visible Cu(2+) sites in pMMO from Methylococcus capsulatus (Bath) and Methylomicrobium album BG8.

    PubMed

    Lemos, S S; Perille Collins, M L; Eaton, S S; Eaton, G R; Antholine, W E

    2000-08-01

    X-band (9.1 GHz) and S-band (3.4 GHz) electron paramagnetic resonance (EPR) spectra for particulate methane monooxygenase (pMMO) in whole cells from Methylococcus capsulatus (Bath) grown on (63)Cu and (15)N were obtained and compared with previously reported spectra for pMMO from Methylomicrobium album BG8. For both M. capsulatus (Bath) and M. album BG8, two nearly identical Cu(2+) EPR signals with resolved hyperfine coupling to four nitrogens are observed. The EPR parameters for pMMO from M. capsulatus (Bath) (g( parallel) = 2.244, A( parallel) = 185 G, and A(N) = 19 G for signal one; g( parallel) = 2.246, A( parallel) = 180 G, and A(N) = 19 G for signal two) and for pMMO from M. album BG8 (g( parallel) = 2.243, A( parallel) = 180 G, and A(N) = 18 G for signal one; g( parallel) = 2. 251, A( parallel) = 180 G, and A(N) = 18 G for signal two) are very similar and are characteristic of type 2 Cu(2+) in a square planar or square pyramidal geometry. In three-pulse electron spin echo envelope modulation (ESEEM) data for natural-abundance samples, nitrogen quadrupolar frequencies due to the distant nitrogens of coordinated histidine imidazoles were observed. The intensities of the quadrupolar combination bands indicate that there are three or four coordinated imidazoles, which implies that most, if not all, of the coordinated nitrogens detected in the continuous wave spectra are from histidine imidazoles. PMID:10920038

  20. Comparison of EPR-visible Cu(2+) sites in pMMO from Methylococcus capsulatus (Bath) and Methylomicrobium album BG8.

    PubMed Central

    Lemos, S S; Perille Collins, M L; Eaton, S S; Eaton, G R; Antholine, W E

    2000-01-01

    X-band (9.1 GHz) and S-band (3.4 GHz) electron paramagnetic resonance (EPR) spectra for particulate methane monooxygenase (pMMO) in whole cells from Methylococcus capsulatus (Bath) grown on (63)Cu and (15)N were obtained and compared with previously reported spectra for pMMO from Methylomicrobium album BG8. For both M. capsulatus (Bath) and M. album BG8, two nearly identical Cu(2+) EPR signals with resolved hyperfine coupling to four nitrogens are observed. The EPR parameters for pMMO from M. capsulatus (Bath) (g( parallel) = 2.244, A( parallel) = 185 G, and A(N) = 19 G for signal one; g( parallel) = 2.246, A( parallel) = 180 G, and A(N) = 19 G for signal two) and for pMMO from M. album BG8 (g( parallel) = 2.243, A( parallel) = 180 G, and A(N) = 18 G for signal one; g( parallel) = 2. 251, A( parallel) = 180 G, and A(N) = 18 G for signal two) are very similar and are characteristic of type 2 Cu(2+) in a square planar or square pyramidal geometry. In three-pulse electron spin echo envelope modulation (ESEEM) data for natural-abundance samples, nitrogen quadrupolar frequencies due to the distant nitrogens of coordinated histidine imidazoles were observed. The intensities of the quadrupolar combination bands indicate that there are three or four coordinated imidazoles, which implies that most, if not all, of the coordinated nitrogens detected in the continuous wave spectra are from histidine imidazoles. PMID:10920038

  1. The presence of multiple c-type cytochromes at the surface of the methanotrophic bacterium Methylococcus capsulatus (Bath) is regulated by copper.

    PubMed

    Karlsen, O A; Lillehaug, J R; Jensen, H B

    2008-10-01

    Identification of surface proteins is essential to understand bacterial communication with its environment. Analysis of the surface-associated proteins of Methylococcus capsulatus (Bath) revealed a highly dynamic structure responding closely to the availability of copper in the medium in the range from approximately 0 to 10 microM. Several c-type cytochromes, including three novel multihaem proteins, are present at the cellular surface, a feature that is otherwise a peculiarity of dissimilatory metal-reducing bacteria. At low copper concentrations, the cytochrome c(553o) and the cytochrome c(553o) family protein, encoded by the MCA0421 and MCA0423 genes, respectively, are major constituents of the surfaceome and show a fine-tuned copper-dependent regulation of expression. Two novel members of the cytochrome c(553o) family were identified: MCA0338 was abundant between 5 and 10 microM copper, while MCA2259 was detected only in the surface fraction obtained from approximately 0 microM copper cultures. The presence at the bacterial surface of several c-type cytochromes, generally involved in energy transduction, indicates strongly that redox processes take place at the bacterial surface. Due to the unique role of copper in the biology of M. capsulatus (Bath), it appears that c-type cytochromes have essential functions in copper homeostasis allowing the cells to adapt to varying copper exposure. PMID:18681943

  2. Development and validation of promoter-probe vectors for the study of methane monooxygenase gene expression in Methylococcus capsulatus Bath.

    PubMed

    Ali, Hanif; Murrell, J Colin

    2009-03-01

    A series of integrative and versatile broad-host-range promoter-probe vectors carrying reporter genes encoding green fluorescent protein (GFP), catechol 2,3-dioxygenase (XylE) or beta-galactosidase (LacZ) were constructed for use in methanotrophs. These vectors facilitated the measurement of in vivo promoter activity in methanotrophs under defined growth conditions. They were tested by constructing transcriptional fusions between the soluble methane monooxygenase (sMMO) sigma(54) promoter or particulate methane monooxygenase (pMMO) sigma(70) promoter from Methylococcus capsulatus and the reporter genes. Reporter gene activity was measured under high- and low-copper growth conditions and the data obtained closely reflected transcriptional regulation of the sMMO or pMMO operon, thus demonstrating the suitability of these vectors for assessing promoter activity in methanotrophs. When beta-galactosidase expression was coupled with the fluorogenic substrate 4-methylumbelliferyl beta-D-glucuronide it yielded a sensitive and powerful screening system for detecting cells expressing this reporter gene. These data were substantiated with independent experiments using RT-PCR and RNA dot-blot analysis. PMID:19246747

  3. Genes involved in the copper-dependent regulation of soluble methane monooxygenase of Methylococcus capsulatus (Bath): cloning, sequencing and mutational analysis.

    PubMed

    Csáki, Róbert; Bodrossy, Levente; Klem, József; Murrell, J Colin; Kovács, Kornél L

    2003-07-01

    The key enzyme in methane metabolism is methane monooxygenase (MMO), which catalyses the oxidation of methane to methanol. Some methanotrophs, including Methylococcus capsulatus (Bath), possess two distinct MMOs. The level of copper in the environment regulates the biosynthesis of the MMO enzymes in these methanotrophs. Under low-copper conditions, soluble MMO (sMMO) is expressed and regulation takes place at the level of transcription. The structural genes of sMMO were previously identified as mmoXYBZ, mmoD and mmoC. Putative transcriptional start sites, containing a sigma(70)- and a sigma(N)-dependent motif, were identified in the 5' region of mmoX. The promoter region of mmoX was mapped using truncated 5' end regions fused to a promoterless green fluorescent protein gene. A 9.5 kb region, adjacent to the sMMO structural gene cluster, was analysed. Downstream (3') from the last gene of the operon, mmoC, four ORFs were found, mmoG, mmoQ, mmoS and mmoR. mmoG shows significant identity to the large subunit of the bacterial chaperonin gene, groEL. In the opposite orientation, two genes, mmoQ and mmoS, showed significant identity to two-component sensor-regulator system genes. Next to mmoS, a gene encoding a putative sigma(N)-dependent transcriptional activator, mmoR was identified. The mmoG and mmoR genes were mutated by marker-exchange mutagenesis and the effects of these mutations on the expression of sMMO was investigated. sMMO transcription was impaired in both mutants. These results indicate that mmoG and mmoR are essential for the expression of sMMO in Mc. capsulatus (Bath). PMID:12855730

  4. Effect of methanobactin on the activity and electron paramagnetic resonance spectra of the membrane-associated methane monooxygenase in Methylococcus capsulatus Bath.

    PubMed

    Choi, Dong W; Antholine, William E; Do, Young S; Semrau, Jeremy D; Kisting, Clint J; Kunz, Ryan C; Campbell, Damon; Rao, Vinay; Hartsel, Scott C; DiSpirito, Alan A

    2005-10-01

    Improvements in the purification of methanobactin (mb) from either Methylosinus trichosporium OB3b(T) or Methylococcus capsulatus Bath resulted in preparations that stimulated methane-oxidation activity in both whole-cell and cell-free fractions of Methylococcus capsulatus Bath expressing the membrane-associated methane monooxygenase (pMMO). By using washed membrane factions with pMMO activities in the 290 nmol propylene oxidized min(-1) (mg protein)(-1) range, activities approaching 400 nmol propylene oxidized min(-1) (mg protein)(-1) were commonly observed following addition of copper-containing mb (Cu-mb), which represented 50-75 % of the total whole-cell activity. The stimulation of methane-oxidation activity by Cu-mb was similar to or greater than that observed with equimolar concentrations of Cu(II), without the inhibitory effects observed with high copper concentrations. Stimulation of pMMO activity was not observed with copper-free mb, nor was it observed when the copper-to-mb ratio was <0.5 Cu atoms per mb. The electron paramagnetic resonance (EPR) spectra of mb differed depending on the copper-to-mb ratio. At copper-to-mb ratios of <0.4 Cu(II) per mb, Cu(II) addition to mb showed an initial coordination by both sulfur and nitrogen, followed by reduction to Cu(I) in <2 min. At Cu(II)-to-mb ratios between 0.4 and 0.9 Cu(II) per mb, the intensity of the Cu(II) signal in EPR spectra was more representative of the Cu(II) added and indicated more nitrogen coordination. The EPR spectral properties of mb and pMMO were also examined in the washed membrane fraction following the addition of Cu(II), mb and Cu-mb in the presence or absence of reductants (NADH or duroquinol) and substrates (CH4 and/or O2). The results indicated that Cu-mb increased electron flow to the pMMO, increased the free radical formed following the addition of O2 and decreased the residual free radical following the addition of O2 plus CH4. The increase in pMMO activity and EPR spectral changes

  5. Purified particulate methane monooxygenase from Methylococcus capsulatus (Bath) is a dimer with both mononuclear copper and a copper-containing cluster

    PubMed Central

    Lieberman, Raquel L.; Shrestha, Deepak B.; Doan, Peter E.; Hoffman, Brian M.; Stemmler, Timothy L.; Rosenzweig, Amy C.

    2003-01-01

    Particulate methane monooxygenase (pMMO) is a membrane-bound enzyme that catalyzes the oxidation of methane to methanol in methanotropic bacteria. Understanding how this enzyme hydroxylates methane at ambient temperature and pressure is of fundamental chemical and potential commercial importance. Difficulties in solubilizing and purifying active pMMO have led to conflicting reports regarding its biochemical and biophysical properties, however. We have purified pMMO from Methylococcus capsulatus (Bath) and detected activity. The purified enzyme has a molecular mass of ≈200 kDa, probably corresponding to an α2β2γ2 polypeptide arrangement. Each 200-kDa pMMO complex contains 4.8 ± 0.8 copper ions and 1.5 ± 0.7 iron ions. Electron paramagnetic resonance spectroscopic parameters corresponding to 40–60% of the total copper are consistent with the presence of a mononuclear type 2 copper site. X-ray absorption near edge spectra indicate that purified pMMO is a mixture of Cu(I) and Cu(II) oxidation states. Finally, extended x-ray absorption fine structure data are best fit with oxygen/nitrogen ligands and a 2.57-Å Cu-Cu interaction, providing direct evidence for a copper-containing cluster in pMMO. PMID:12634423

  6. The hydroxylase component of soluble methane monooxygenase from Methylococcus capsulatus (Bath) exists in several forms as shown by electrospray-ionisation mass spectrometry.

    PubMed

    Buzy, A; Millar, A L; Legros, V; Wilkins, P C; Dalton, H; Jennings, K R

    1998-06-15

    The hydroxylase of the soluble methane monooxygenase from the bacterium Methylococcus capsulatus (Bath) has been investigated by means of electrospray-ionisation mass spectrometry (ESI-MS) and liquid chromatography ESI-MS (LC/ESI-MS). The hydroxylase is a non-heme diiron protein consisting of three pairs of non-identical subunits (alpha approximately 60 kDa, beta approximately 45 kDa and gamma approximately 20 kDa). Liquid chromatographic separation of the hydroxylase subunits was required before MS analysis in order to detect the alpha-subunit. The masses measured for the three subunits were found to disagree with those calculated from their gene sequences. Experiments involving the use of CNBr and trypsin cleavage followed by LC/ESI-MS and MS/MS analyses permitted the location and correction of errors in the sequences deduced from the use of cDNA. The ESI-MS results also showed that the alpha-subunit of the hydroxylase exists in multiple forms which result from cleavage of the protein. This observation explains a number of enigmatic features of the protein previously reported in the literature and illustrates the pivotal role of ESI-MS in complementing data obtained from molecular biology for the characterisation of the primary sequence of proteins. PMID:9688272

  7. Identification of putative methanol dehydrogenase (moxF) structural genes in methylotrophs and cloning of moxF genes from Methylococcus capsulatus bath and Methylomonas albus BG8.

    PubMed Central

    Stephens, R L; Haygood, M G; Lidstrom, M E

    1988-01-01

    An open-reading-frame fragment of a Methylobacterium sp. strain AM1 gene (moxF) encoding a portion of the methanol dehydrogenase structural protein has been used as a hybridization probe to detect similar sequences in a variety of methylotrophic bacteria. This hybridization was used to isolate clones containing putative moxF genes from two obligate methanotrophic bacteria, Methylococcus capsulatus Bath and Methylomonas albus BG8. The identity of these genes was confirmed in two ways. A T7 expression vector was used to produce methanol dehydrogenase protein in Escherichia coli from the cloned genes, and in each case the protein was identified by immunoblotting with antiserum against the Methylomonas albus methanol dehydrogenase. In addition, a moxF mutant of Methylobacterium strain AM1 was complemented to a methanol-positive phenotype that partially restored methanol dehydrogenase activity, using broad-host-range plasmids containing the moxF genes from each methanotroph. The partial complementation of a moxF mutant in a facultative serine pathway methanol utilizer by moxF genes from type I and type X obligate methane utilizers suggests broad functional conservation of the methanol oxidation system among gram-negative methylotrophs. Images PMID:3129400

  8. Purified particulate methane monooxygenase from Methylococcus capsulatus (Bath) is a dimer with both mononuclear copper and a copper-containing cluster.

    PubMed

    Lieberman, Raquel L; Shrestha, Deepak B; Doan, Peter E; Hoffman, Brian M; Stemmler, Timothy L; Rosenzweig, Amy C

    2003-04-01

    Particulate methane monooxygenase (pMMO) is a membrane-bound enzyme that catalyzes the oxidation of methane to methanol in methanotropic bacteria. Understanding how this enzyme hydroxylates methane at ambient temperature and pressure is of fundamental chemical and potential commercial importance. Difficulties in solubilizing and purifying active pMMO have led to conflicting reports regarding its biochemical and biophysical properties, however. We have purified pMMO from Methylococcus capsulatus (Bath) and detected activity. The purified enzyme has a molecular mass of approximately 200 kDa, probably corresponding to an alpha(2)beta(2)gamma(2) polypeptide arrangement. Each 200-kDa pMMO complex contains 4.8 +/- 0.8 copper ions and 1.5 +/- 0.7 iron ions. Electron paramagnetic resonance spectroscopic parameters corresponding to 40-60% of the total copper are consistent with the presence of a mononuclear type 2 copper site. X-ray absorption near edge spectra indicate that purified pMMO is a mixture of Cu(I) and Cu(II) oxidation states. Finally, extended x-ray absorption fine structure data are best fit with oxygennitrogen ligands and a 2.57-A Cu-Cu interaction, providing direct evidence for a copper-containing cluster in pMMO. PMID:12634423

  9. The C-terminal part of the surface-associated protein MopE of the methanotroph Methylococcus capsulatus (Bath) is secreted into the growth medium.

    PubMed

    Fjellbirkeland, A; Kruger, P G; Bemanian, V; Høgh, B T; Murrell, J C; Jensen, H B

    2001-09-01

    A protein with an apparent molecular mass of 46 kDa was detected as the major polypeptide in the culture medium of the biotechnologically important methanotrophic bacterium Methylococcus capsulatus (Bath). The protein cross-reacted with polyclonal antibodies raised against the outer-membrane-associated protein MopE. The antiserum was used to identify a positive clone from a lambda gt11 library. The nucleotide sequence determined for the clone demonstrated that MopE and the secreted protein are encoded by the same gene, and that the secreted protein represents an N-terminally truncated form of MopE. By using monospecific antibodies against MopE in immunogold electron microscopy, the protein was localized at the cell surface and cell periphery. The mopE gene was expressed in Escherichia coli. The MopE protein synthesized was found in the periplasmic space of E. coli. No protein with sequence similarity over the entire length of MopE was detected in the databases, but some sequence similarity to the copper-repressible CorA protein of the methanotroph Methylomicrobium albus (Berson and Lidstrom 1997) was observed for the C-terminal region of MopE. PMID:11511867

  10. A low-molecular-mass protein from Methylococcus capsulatus (Bath) is responsible for the regulation of formaldehyde dehydrogenase activity in vitro.

    PubMed

    Tate, S; Dalton, H

    1999-01-01

    An 8.6 kDa protein, which the authors call a modifin, has been purified from Methylococcus capsulatus (Bath) and has been shown to alter the substrate specificity and kinetics of NAD+-linked formaldehyde dehydrogenase (FDH) isolated from the same organism. Purification methods for both the modifin and FDH are presented which reliably produced pure protein for further analysis. Analysis of the molecular mass and N-terminal sequence of both FDH and the modifin indicate that they are unique proteins and show no similarity to alcohol or aldehyde dehydrogenase enzymes isolated from methylotrophic bacteria. Substrate specificity studies demonstrated that FDH oxidized formaldehyde exclusively in the presence of the modifin; a diverse range of aldehydes and alcohols were oxidized by FDH in the absence of the modifin. No formaldehyde oxidation was detected in the absence of the modifin. Attempts to replace the modifin with glutathione or high concentrations of methanol to stimulate formaldehyde oxidation failed. With acetaldehyde as substrate, FDH showed standard Michaelis-Menten kinetics; interaction of FDH with the modifin using formaldehyde as substrate altered the kinetics of the reaction to sigmoidal. Kinetic analysis during turnover experiments indicated that the FDH may be associated with bound formaldehyde following enzyme isolation and that NAD may also be associated with the enzyme but in a form that is less tightly bound than found with the methanol dehydrogenase from Bacillus methanolicus. Data are presented which indicate that the modifin may play an important role in regulating formaldehyde concentration in vivo. PMID:10206695

  11. Molecular analysis of an outer membrane protein, MopB, of Methylococcus capsulatus (Bath) and structural comparisons with proteins of the OmpA family.

    PubMed

    Fjellbirkeland, A; Bemanian, V; McDonald, I R; Murrell, J C; Jensen, H B

    2000-01-01

    The gene encoding a major outer membrane protein (MopB) of the methanotroph Methylococcus capsulatus (Bath) was cloned and sequenced. The cloned DNA contained an open reading frame of 1044 bp coding for a 348-amino-acid polypeptide with a 21-amino-acid leader peptide. Comparative sequence analysis of the predicted amino acid sequence revealed that the C-terminal part of MopB possessed sequences that are conserved in the OmpA family of proteins. The N-terminal half of the protein had no significant sequence similarity to other proteins in the databases, but the predicted secondary structure showed stretches of amphipathic beta-strands typical of transmembrane segments of outer membrane proteins. A region with four cysteines similar to the cysteine-encompassing region of the OprF of Pseudomonas aeruginosa was found toward the C-terminal part of MopB. Results from whole-cell labeling with the fluorescent thiol-reacting reagent 5-iodoacetamidofluorescein indicated a surface-exposed location for these cysteines. A probe consisting of the 3'-end of the mopB gene hybridized to the type I methanotroph Methylomonas methanica S in Southern blots containing DNA from nine methanotrophic strains representing six different genera. PMID:10896213

  12. The soluble methane mono-oxygenase of Methylococcus capsulatus (Bath). Its ability to oxygenate n-alkanes, n-alkenes, ethers, and alicyclic, aromatic and heterocyclic compounds.

    PubMed Central

    Colby, J; Stirling, D I; Dalton, H

    1977-01-01

    1. Methane mono-oxygenase of Methylococcus capsulatus (Bath) catalyses the oxidation of various substituted methane derivatives including methanol. 2. It is a very non-specific oxygenase and, in some of its catalytic properties, apparently resembles the analogous enzyme from Methylomonas methanica but differs from those found in Methylosinus trichosporium and Methylomonas albus. 3. CO is oxidized to CO2. 4. C1-C8 n-alkanes are hydroxylated, yielding mixtures of the corresponding 1- and 2-alcohols; no 3- or 4-alcohols are formed. 5. Terminal alkenes yield the corresponding 1,2-epoxides. cis- or trans-but-2-ene are each oxidized to a mixture of 2,3-epoxybutane and but-2-en-1-ol with retention of the cis or trans configuration in both products; 2-butanone is also formed from cis-but-2-ene only. 6. Dimethyl ether is oxidized. Diethyl ether undergoes sub-terminal oxidation, yielding ethanol and ethanal in equimolar amounts. 7. Methane mono-oxygenase also hydroxylates cyclic alkanes and aromatic compounds. However, styrene yields only styrene epoxide and pyridine yields only pyridine N-oxide. 8. Of those compounds tested, only NADPH can replace NADH as electron donor. PMID:411486

  13. Regulation of bacterial methane oxidation: transcription of the soluble methane mono-oxygenase operon of Methylococcus capsulatus (Bath) is repressed by copper ions.

    PubMed

    Nielsen, A K; Gerdes, K; Degn, H; Murrell, J C

    1996-05-01

    Methane is oxidized to methanol by the enzyme methane mono-oxygenase (MMO) in methanotrophic bacteria. In previous work, this multicomponent enzyme system has been extensively characterized at the biochemical and molecular level. Copper ions have been shown to irreversibly inhibit MMO activity in vivo and in vitro, but the effect of copper ions on transcription of the genes encoding the soluble (cytoplasmic) MMO (sMMO) has not previously been investigated. To examine more closely the regulation of bacterial methane oxidation and to determine the role of copper in this process, we have investigated transcriptional regulation of the sMMO gene cluster in the methanotrophic bacterium Methylococcus capsulatus (Bath). Using Northern blot analysis and primer extension experiments, it was shown that the six ORFs of the sMMO gene cluster are organized as an operon and the transcripts produced upon expression of this operon have been identified. The synthesis of these transcripts was under control of a single copper-regulated promoter, which is as yet not precisely defined. PMID:8704968

  14. Resolution of the methane mono-oxygenase of Methylococcus capsulatus (Bath) into three components. Purification and properties of component C, a flavoprotein.

    PubMed Central

    Colby, J; Dalton, H

    1978-01-01

    1. Ion-exchange chromatography resolves the methane mono-oxygenase from soluble extracts of Methylococcus capsulatus (Bath) into three fractions. 2. Fractions A and B are comparatively stable at 0 degrees C, whereas fraction C is very unstable unless kept in the presence of sodium thioglycollate (1-10 mM) or dithiothreitol (5-10mM). 3. The active component from fraction C was purified some 80-fold. 4. Purified component C has mol. wt. 42000. Its solutions are yellow with absorption maxima at 270 and 465 nm and a shoulder at 395 nm. The 465 nm peak is abolished by reduction with NADH or sodium dithionite, or by photoreduction in the presence of EDTA. A new spectral species, probably a neutral flavin semiquinone, is observed on partial reduction of component C. 5. No copper was detected in samples of purified component C, but the protein contains 1.3-1.5 atoms of iron/molecule. 6. On boiling, component C releases a yellow-green fluorescent material that has been identified as FAD from its absorption and fluorescence spectra and by t.l.c. 7. Component C contains 1 mol of FAD/mol of protein. PMID:418777

  15. Identification of putative methanol dehydrogenase (moxF) structural genes in methylotrophs and cloning of moxF genes from methylococcus capsulatus bath and Methylomonas albus BG8

    SciTech Connect

    Stephens, R.L.; Haygood, M.G.; Lidstrom, M.E.

    1988-05-01

    An open-reading-frame fragment of a Methylobacterium sp. strain AM1 gene (moxF) encoding a portion of the methanol dehydrogenase structural protein has been used as a hybridization probe to detect similar sequences in a variety of methylotrophic bacteria. This hybridization was used to isolate clones containing putative moxF genes from two obligate methanotrophic bacteria, Methylococcus capsulatus Bath and Methylomonas albus BG8. The identity of these genes was confirmed in two ways. A T7 expression vector was used to produce methanol dehydrogenase protein in Escherichia coli from the cloned genes,a and in each case the protein was identified by immunoblotting with antiserum against the Methylomonas albus methanol dehydrogenase. In addition, a moxF mutant of Methylobacterium strain AM1 was complemented to a methanol-positive phenotype that partially restored methanol dehydrogenase activity, using broad-host-range plasmids containing the moxF genes from each methanotroph. The partial complementation of a moxF mutant in a facultative serine pathway methanol utilizer by moxF genes from type I and type X obligate methane utilizers suggests broad functional conservation of the methanol oxidation system among gram-negative methylotrophs.

  16. Crystal structures of the soluble methane monooxygenase hydroxylase from Methylococcus capsulatus (Bath) demonstrating geometrical variability at the dinuclear iron active site.

    PubMed

    Whittington, D A; Lippard, S J

    2001-02-01

    The oxidation of methane to methanol is performed at carboxylate-bridged dinuclear iron centers in the soluble methane monooxygenase hydroxylase (MMOH). Previous structural studies of MMOH, and the related R2 subunit of ribonucleotide reductase, have demonstrated the occurrence of carboxylate shifts involving glutamate residues that ligate the catalytic iron atoms. These shifts are thought to have important mechanistic implications. Recent kinetic and theoretical studies have also emphasized the importance of hydrogen bonding and pH effects at the active site. We report here crystal structures of MMOH from Methylococcus capsulatus (Bath) in the diiron(II), diiron(III), and mixed-valent Fe(II)Fe(III) oxidation states, and at pH values of 6.2, 7.0, and 8.5. These structures were investigated in an effort to delineate the range of possible motions at the MMOH active site and to identify hydrogen-bonding interactions that may be important in understanding catalysis by the enzyme. Our results present the first view of the diiron center in the mixed-valent state, and they indicate an increased lability for ferrous ions in the enzyme. Alternate conformations of Asn214 near the active site according to redox state and a distortion in one of the alpha-helices adjacent to the metal center in the diiron(II) state have also been identified. These changes alter the surface of the protein in the vicinity of the catalytic core and may have implications for small-molecule accessibility to the active site and for protein component interactions in the methane monooxygenase system. Collectively, these results help to explain previous spectroscopic observations and provide new insight into catalysis by the enzyme. PMID:11456616

  17. Positively charged amino acids are essential for electron transfer and protein-protein interactions in the soluble methane monooxygenase complex from Methylococcus capsulatus (Bath).

    PubMed

    Balendra, Suki; Lesieur, Claire; Smith, Thomas J; Dalton, Howard

    2002-02-26

    The soluble methane monooxygenase (sMMO) complex from Methylococcus capsulatus (Bath) catalyses oxygen- and NAD(P)H-dependent oxygenation of methane, propene, and other substrates. Whole-complex sMMO oxygenase activity requires all three sMMO components: the hydroxylase, the reductase, and protein B. Also, in the presence of hydrogen peroxide, the hydroxylase alone catalyzes substrate oxygenation via the peroxide shunt reaction. We investigated the effect of amine cross-linking on hydroxylase activity to probe the role of a gross conformational change that occurs in the hydroxylase upon binding of the other protein components. The cross-linker inhibited hydroxylase activity in the whole complex, but this effect was due to covalent modification of primary amine groups rather than cross-linking. Covalent modification of arginine side-chains on the hydroxylase had a similar effect, but, most remarkably, neither form of modification affected the activity of the hydroxylase via the peroxide shunt reaction. It was shown that covalent modification of positively charged groups on the hydroxylase, which occurred at multiple sites, interfered with its physical and functional interactions with protein B and with the passage of electrons from the reductase. These results indicate that protein B and the reductase of the sMMO complex interact via positively charged groups on the surface of the hydroxylase to induce a conformational change that is necessary for delivery of electrons into the active site of the hydroxylase. Modification of positively charged groups on protein B had no effect on its function, consistent with the hypothesis that positively charged groups on the hydroxylase interact with negative charges on protein B. Thus, we have discovered a means of specifically inactivating the interactions between the sMMO complex while preserving the catalytic activity of the hydroxylase active site which provides a new method of studying intercomponent interactions within s

  18. Inactivation of the regulatory protein B of soluble methane monooxygenase from Methylococcus capsulatus (Bath) by proteolysis can be overcome by a Gly to Gln modification.

    PubMed

    Lloyd, J S; Bhambra, A; Murrell, J C; Dalton, H

    1997-08-15

    The regulatory protein B of soluble methane monooxygenase (sMMO) from Methylococcus capsulatus (Bath), exists as a mixture of the full-length active form and truncated forms, B' and B". Electrospray ionisation mass spectrometry (ESI-MS) was used to identify a cleavage site between Met12 and Gly13, such that 12 amino acids were lost from the N-terminus of protein B. This truncate was designated B' and molecular masses were assigned to proteins B and B' of 15,852.6+/-0.4 Da and 14,629.5+/-0.3 Da, respectively. A cleavage site between Gln29 and Val30 was also identified such that 29 amino acids were lost from the N-terminus of protein B. This truncate was designated B" and had a molecular mass of 12,709.93+/-0.02 Da. Proteins B' and B" were found to be inactive in the sMMO system. Addition of protease inhibitors or the heterologous expression of protein B in various strains of lon-deficient or ompT-deficient Escherichia coli, did not inhibit B' formation. Expression of protein B as a glutathione S-transferase fusion protein and subsequent purification of protein B from E. coli using affinity chromatography resulted in preparations of protein B with higher enzyme activities than that of wild-type protein B. However, ESI-MS confirmed that protein B' was still present. Alteration of the Met12-Gly13 cleavage site to Met12-Gln13 revealed that the stability of G13Q at 20 degrees C and 37 degrees C was higher than that of wild-type preparations. ESI-MS indicated that protein B' was absent and could only be identified after prolonged incubation at room temperature. The amount of active protein B present in the cell may be controlled by protein B cleavage, thereby regulating electron transfer. Alternatively, it may allow protein B to maintain a certain conformation necessary for enzyme activity and this may control the activity of sMMO in response to the supply of methane to the cell. PMID:9310362

  19. The stereospecific hydroxylation of [2,2-2H2]butane and chiral dideuteriobutanes by the particulate methane monooxygenase from Methylococcus capsulatus (Bath).

    PubMed

    Yu, Steve S-F; Wu, Lo-Ying; Chen, Kelvin H-C; Luo, Wen-I; Huang, Ded-Shih; Chan, Sunney I

    2003-10-17

    Experiments on cryptically chiral ethanes have indicated that the particulate methane monooxygenase (pMMO) from Methylococcus capsulatus (Bath) catalyzes the hydroxylation of ethane with total retention of configuration at the carbon center attacked. This result would seem to rule out a radical mechanism for the hydroxylation chemistry, at least as mediated by this enzyme. The interpretation of subsequent experiments on n-propane, n-butane, and n-pentane has been complicated by hydroxylation at both the pro-R and pro-S secondary C-H bonds, where the hydroxylation takes place. It has been suggested that these results merely reflect presentation of both the pro-R and pro-S C-H bonds to the hot "oxygen atom" species generated at the active site, and that the oxo-transfer chemistry, in fact, proceeds concertedly with retention of configuration. In the present work, we have augmented these earlier studies with experiments on [2,2-2H2]butane and designed d,l form chiral dideuteriobutanes. Essentially equal amounts of (2R)-[3,3-2H2]butan-2-ol and (2R)-[2-2H1]butan-2-ol are produced upon hydroxylation of [2,2-2H2]butane. The chemistry is stereospecific with full retention of configuration at the secondary carbon oxidized. In the case of the various chiral deuterated butanes, the extent of configurational inversion has been shown to be negligible for all the chiral butanes examined. Thus, the hydroxylation of butane takes place with full retention of configuration in butane as well as in the case of ethane. These results are interpreted in terms of an oxo-transfer mechanism based on side-on singlet oxene insertion across the C-H bond similar to that previously noted for singlet carbene insertion (Kirmse, W., and Ozkir, I. S. (1992) J. Am. Chem. Soc. 114, 7590-7591). Finally, we discuss how even the oxene insertion mechanism, with "spin crossover" in the transition state, could lead to small amounts of radical rearrangement products, if and when such products are observed. A

  20. Production of High-Quality Particulate Methane Monooxygenase in High Yields from Methylococcus capsulatus (Bath) with a Hollow-Fiber Membrane Bioreactor

    PubMed Central

    Yu, Steve S.-F.; Chen, Kelvin H.-C.; Tseng, Mandy Y.-H.; Wang, Yane-Shih; Tseng, Chiu-Feng; Chen, Yu-Ju; Huang, Ded-Shih; Chan, Sunney I.

    2003-01-01

    In order to obtain particulate methane monooxygenase (pMMO)-enriched membranes from Methylococcus capsulatus (Bath) with high activity and in high yields, we devised a method to process cell growth in a fermentor adapted with a hollow-fiber bioreactor that allows easy control and quantitative adjustment of the copper ion concentration in NMS medium over the time course of cell culture. This technical improvement in the method for culturing bacterial cells allowed us to study the effects of copper ion concentration in the growth medium on the copper content in the membranes, as well as the specific activity of the enzyme. The optimal copper concentration in the growth medium was found to be 30 to 35 μM. Under these conditions, the pMMO is highly expressed, accounting for 80% of the total cytoplasmic membrane proteins and having a specific activity as high as 88.9 nmol of propylene oxide/min/mg of protein with NADH as the reductant. The copper stoichiometry is ∼13 atoms per pMMO molecule. Analysis of other metal contents provided no evidence of zinc, and only traces of iron were present in the pMMO-enriched membranes. Further purification by membrane solubilization in dodecyl β-d-maltoside followed by fractionation of the protein-detergent complexes according to molecular size by gel filtration chromatography resulted in a good yield of the pMMO-detergent complex and a high level of homogeneity. The pMMO-detergent complex isolated in this way had a molecular mass of 220 kDa and consisted of an αβγ protein monomer encapsulated in a micelle consisting of ca. 240 detergent molecules. The enzyme is a copper protein containing 13.6 mol of copper/mol of pMMO and essentially no iron (ratio of copper to iron, 80:1). Both the detergent-solubilized membranes and the purified pMMO-detergent complex exhibited reasonable, if not excellent, specific activity. Finally, our ability to control the level of expression of the pMMO allowed us to clarify the sensitivity of the

  1. The particulate methane monooxygenase from methylococcus capsulatus (Bath) is a novel copper-containing three-subunit enzyme. Isolation and characterization.

    PubMed

    Nguyen, H H; Elliott, S J; Yip, J H; Chan, S I

    1998-04-01

    The particulate methane monooxygenase (pMMO) is known to be very difficult to study mainly due to its unusual activity instability in vitro. By cultivating Methylococcus capsulatus (Bath) under methane stress conditions and high copper levels in the growth medium, membranes highly enriched in the pMMO with exceptionally stable activity can be isolated from these cells. Purified and active pMMO can be subsequently obtained from these membrane preparations using protocols in which an excess of reductants and anaerobic conditions were maintained during membrane solubilization by dodecyl beta-D-maltoside and purification by chromatography. The pMMO was found to be the major constituent in these membranes, constituting 60-80% of total membrane proteins. The dominant species of the pMMO was found to consist of three subunits, alpha, beta, and gamma, with an apparent molecular mass of 45, 26, and 23 kDa, respectively. A second species of the pMMO, a proteolytically processed version of the enzyme, was found to be composed of three subunits, alpha', beta, and gamma, with an apparent molecular mass of 35, 26, and 23 kDa, respectively. The alpha and alpha' subunits from these two forms of the pMMO contain identical N-terminal sequences. The gamma subunit, however, exhibits variation in its N-terminal sequence. The pMMO is a copper-containing protein only and shows a requirement for Cu(I) ions. Approximately 12-15 Cu ions per 94-kDa monomeric unit were observed. The pMMO is sensitive to dioxygen tension. On the basis of dioxygen sensitivity, three kinetically distinct forms of the enzyme can be distinguished. A slow but air-stable form, which is converted into a "pulsed" state upon direct exposure to atmospheric oxygen pressure, is considered as type I pMMO. This form was the subject of our pMMO isolation effort. Other forms (types II and III) are deactivated to various extents upon exposure to atmospheric dioxygen pressure. Under inactivating conditions, these unstable forms

  2. Expression and characterization of ferredoxin and flavin adenine dinucleotide binding domains of the reductase component of soluble methane monooxygenase from Methylococcus capsulatus (Bath).

    PubMed

    Blazyk, Jessica L; Lippard, Stephen J

    2002-12-31

    Soluble methane monooxygenase (sMMO) from Methylococcus capsulatus (Bath) catalyzes the selective oxidation of methane to methanol, the first step in the primary catabolic pathway of methanotrophic bacteria. A reductase (MMOR) mediates electron transfer from NADH through its FAD and [2Fe-2S] cofactors to the dinuclear non-heme iron sites housed in a hydroxylase (MMOH). The structurally distinct [2Fe-2S], FAD, and NADH binding domains of MMOR facilitated division of the protein into its functional ferredoxin (MMOR-Fd) and FAD/NADH (MMOR-FAD) component domains. The 10.9 kDa MMOR-Fd (MMOR residues 1-98) and 27.6 kDa MMOR-FAD (MMOR residues 99-348) were expressed and purified from recombinant Escherichia coli systems. The Fd and FAD domains have absorbance spectral features identical to those of the [2Fe-2S] and flavin components, respectively, of MMOR. Redox potentials, determined by reductive titrations that included indicator dyes, for the [2Fe-2S] and FAD cofactors in the domains are as follows: -205.2 +/- 1.3 mV for [2Fe-2S](ox/red), -172.4 +/- 2.0 mV for FAD(ox/sq), and -266.4 +/- 3.5 mV for FAD(sq/hq). Kinetic and spectral properties of intermediates observed in the reaction of oxidized MMOR-FAD (FAD(ox)) with NADH at 4 degrees C were established with stopped-flow UV-visible spectroscopy. Analysis of the influence of pH on MMOR-FAD optical spectra, redox potentials, and NADH reaction kinetics afforded pK(a) values for the semiquinone (FAD(sq)) and hydroquinone (FAD(hq)) MMOR-FAD species and two protonatable groups near the flavin cofactor. Electron transfer from MMOR-FAD(hq) to oxidized MMOR-Fd is extremely slow (k = 1500 M(-1) s(-1) at 25 degrees C, compared to 90 s(-1) at 4 degrees C for internal electron transfer between cofactors in MMOR), indicating that cofactor proximity is essential for efficient interdomain electron transfer. PMID:12501207

  3. Production of high-quality particulate methane monooxygenase in high yields from Methylococcus capsulatus (bath) with a hollow-fiber membrane bioreactor.

    PubMed

    Yu, Steve S-F; Chen, Kelvin H-C; Tseng, Mandy Y-H; Wang, Yane-Shih; Tseng, Chiu-Feng; Chen, Yu-Ju; Huang, Ded-Shih; Chan, Sunney I

    2003-10-01

    In order to obtain particulate methane monooxygenase (pMMO)-enriched membranes from Methylococcus capsulatus (Bath) with high activity and in high yields, we devised a method to process cell growth in a fermentor adapted with a hollow-fiber bioreactor that allows easy control and quantitative adjustment of the copper ion concentration in NMS medium over the time course of cell culture. This technical improvement in the method for culturing bacterial cells allowed us to study the effects of copper ion concentration in the growth medium on the copper content in the membranes, as well as the specific activity of the enzyme. The optimal copper concentration in the growth medium was found to be 30 to 35 micro M. Under these conditions, the pMMO is highly expressed, accounting for 80% of the total cytoplasmic membrane proteins and having a specific activity as high as 88.9 nmol of propylene oxide/min/mg of protein with NADH as the reductant. The copper stoichiometry is approximately 13 atoms per pMMO molecule. Analysis of other metal contents provided no evidence of zinc, and only traces of iron were present in the pMMO-enriched membranes. Further purification by membrane solubilization in dodecyl beta-D-maltoside followed by fractionation of the protein-detergent complexes according to molecular size by gel filtration chromatography resulted in a good yield of the pMMO-detergent complex and a high level of homogeneity. The pMMO-detergent complex isolated in this way had a molecular mass of 220 kDa and consisted of an alphabetagamma protein monomer encapsulated in a micelle consisting of ca. 240 detergent molecules. The enzyme is a copper protein containing 13.6 mol of copper/mol of pMMO and essentially no iron (ratio of copper to iron, 80:1). Both the detergent-solubilized membranes and the purified pMMO-detergent complex exhibited reasonable, if not excellent, specific activity. Finally, our ability to control the level of expression of the pMMO allowed us to clarify

  4. Identification of a copper-repressible C-type heme protein of Methylococcus capsulatus (Bath). A member of a novel group of the bacterial di-heme cytochrome c peroxidase family of proteins.

    PubMed

    Karlsen, Odd A; Kindingstad, Louise; Angelskår, Solveig M; Bruseth, Live J; Straume, Daniel; Puntervoll, Pål; Fjellbirkeland, Anne; Lillehaug, Johan R; Jensen, Harald B

    2005-12-01

    Genomic sequencing of the methanotrophic bacterium, Methylococcus capsulatus (Bath), revealed an open reading frame (MCA2590) immediately upstream of the previously described mopE gene (MCA2589). Sequence analyses of the deduced amino acid sequence demonstrated that the MCA2590-encoded protein shared significant, but restricted, sequence similarity to the bacterial di-heme cytochrome c peroxidase (BCCP) family of proteins. Two putative C-type heme-binding motifs were predicted, and confirmed by positive heme staining. Immunospecific recognition and biotinylation of whole cells combined with MS analyses confirmed expression of MCA2590 in M. capsulatus as a protein noncovalently associated with the cellular surface of the bacterium exposed to the cell exterior. Similar to MopE, expression of MCA2590 is regulated by the bioavailability of copper and is most abundant in M. capsulatus cultures grown under low copper conditions, thus indicating an important physiological role under these growth conditions. MCA2590 is distinguished from previously characterized members of the BCCP family by containing a much longer primary sequence that generates an increased distance between the two heme-binding motifs in its primary sequence. Furthermore, the surface localization of MCA2590 is in contrast to the periplasmic location of the reported BCCP members. Based on our experimental and bioinformatical analyses, we suggest that MCA2590 is a member of a novel group of bacterial di-heme cytochrome c peroxidases not previously characterized. PMID:16336269

  5. Cytochrome P460 Genes from the Methanotroph Methylococcus capsulatus Bath†

    PubMed Central

    Bergmann, David J.; Zahn, James A.; Hooper, Alan B.; DiSpirito, Alan A.

    1998-01-01

    P460 cytochromes catalyze the oxidation of hydroxylamine to nitrite. They have been isolated from the ammonia-oxidizing bacterium Nitrosomonas europaea (R. H. Erickson and A. B. Hooper, Biochim. Biophys. Acta 275:231–244, 1972) and the methane-oxidizing bacterium Methylococcus capsulatus Bath (J. A. Zahn et al., J. Bacteriol. 176:5879–5887, 1994). A degenerate oligonucleotide probe was synthesized based on the N-terminal amino acid sequence of cytochrome P460 and used to identify a DNA fragment from M. capsulatus Bath that contains cyp, the gene encoding cytochrome P460. cyp is part of a gene cluster that contains three open reading frames (ORFs), the first predicted to encode a 59,000-Da membrane-bound polypeptide, the second predicted to encode a 12,000-Da periplasmic protein, and the third (cyp) encoding cytochrome P460. The products of the first two ORFs have no apparent similarity to any proteins in the GenBank database. The overall sequence similarity of the P460 cytochromes from M. capsulatus Bath and N. europaea was low (24.3% of residues identical), although short regions of conserved residues are present in the two proteins. Both cytochromes have a C-terminal, c-heme binding motif (CXXCH) and a conserved lysine residue (K61) that may provide an additional covalent cross-link to the heme (D. M. Arciero and A. B. Hooper, FEBS Lett. 410:457–460, 1997). Gene probing using cyp indicated that a cytochrome P460 similar to that from M. capsulatus Bath may be present in the type II methanotrophs Methylosinus trichosporium OB3b and Methylocystis parvus OBBP but not in the type I methanotrophs Methylobacter marinus A45, Methylomicrobium albus BG8, and Methylomonas sp. strains MN and MM2. Immunoblot analysis with antibodies against cytochrome P460 from M. capsulatus Bath indicated that the expression level of cytochrome P460 was not affected either by expression of the two different methane monooxygenases or by addition of ammonia to the culture medium. PMID

  6. The copper responding surfaceome of Methylococccus capsulatus Bath.

    PubMed

    Karlsen, Odd A; Larsen, Oivind; Jensen, Harald B

    2011-10-01

    Proteins on the cellular surface of a bacterium, its surfaceome, are part of the interface between the bacterium and its environment, and are essential for the cells response to its habitat. Methylococcus capsulatus Bath is one of the most extensively studied methane-oxidizers and is considered as a model-methanotroph. The composition of proteins of the surfaceome of M. capsulatus Bath varies with the availability of copper and changes significantly upon only minor changes of copper concentration in the sub-μM concentration range. Proteins that respond to the changes in copper availability include the assumed copper acquisition protein MopE, c-type heme proteins (SACCP, cytochrome c(553o) proteins) and several proteins of unknown function. The most intriguing observation is that multi-heme c-type cytochromes are major constituents of the M. capsulatus Bath surfaceome. This is not commonly observed in bacteria, but is a feature shared with the dissimilatory metal-reducing bacteria. Their presence on the M. capsulatus Bath cellular surface may be linked to the cells ability to efficiently adapt to changing growth conditions and environmental challenges. However, their possible role(s) in methane oxidation, nitrogen metabolism, copper acquisition, redox-reactions and/or electron transport remain(s) at present an open question. This review will discuss the possible significance of these findings. PMID:22092708

  7. Delta8(14)-steroids in the bacterium Methylococcus capsulatus.

    PubMed Central

    Bouvier, P; Rohmer, M; Benveniste, P; Ourisson, G

    1976-01-01

    The 4,4-dimethyl and 4alpha-methyl sterols of the bacterium Methylococcus capsulatus were identified as 4,4-dimethyl- and 4alpha-methyl-5alpha-cholest-8(14)-en-3beta-ol and 4,4-dimethyl- and 4alpha-methyl-5alpha-cholesta-8(14),24-dien-3beta-ol. Sterol biosynthesis is blocked at the level of 4alpha-methyl delta8(14)-sterols. PMID:999649

  8. The Membrane-Associated Methane Monooxygenase (pMMO) and pMMO-NADH:Quinone Oxidoreductase Complex from Methylococcus capsulatus Bath

    PubMed Central

    Choi, Dong-W.; Kunz, Ryan C.; Boyd, Eric S.; Semrau, Jeremy D.; Antholine, William E.; Han, J.-I.; Zahn, James A.; Boyd, Jeffrey M.; de la Mora, Arlene M.; DiSpirito, Alan A.

    2003-01-01

    Improvements in purification of membrane-associated methane monooxygenase (pMMO) have resulted in preparations of pMMO with activities more representative of physiological rates: i.e., >130 nmol · min−1 · mg of protein−1. Altered culture and assay conditions, optimization of the detergent/protein ratio, and simplification of the purification procedure were responsible for the higher-activity preparations. Changes in the culture conditions focused on the rate of copper addition. To document the physiological events that occur during copper addition, cultures were initiated in medium with cells expressing soluble methane monooxygenase (sMMO) and then monitored for morphological changes, copper acquisition, fatty acid concentration, and pMMO and sMMO expression as the amended copper concentration was increased from 0 (approximately 0.3 μM) to 95 μM. The results demonstrate that copper not only regulates the metabolic switch between the two methane monooxygenases but also regulates the level of expression of the pMMO and the development of internal membranes. With respect to stabilization of cell-free pMMO activity, the highest cell-free pMMO activity was observed when copper addition exceeded maximal pMMO expression. Optimization of detergent/protein ratios and simplification of the purification procedure also contributed to the higher activity levels in purified pMMO preparations. Finally, the addition of the type 2 NADH:quinone oxidoreductase complex (NADH dehydrogenase [NDH]) from M. capsulatus Bath, along with NADH and duroquinol, to enzyme assays increased the activity of purified preparations. The NDH and NADH were added to maintain a high duroquinol/duroquinone ratio. PMID:13129946

  9. The membrane-associated methane monooxygenase (pMMO) and pMMO-NADH:quinone oxidoreductase complex from Methylococcus capsulatus Bath.

    PubMed

    Choi, Dong-W; Kunz, Ryan C; Boyd, Eric S; Semrau, Jeremy D; Antholine, William E; Han, J-I; Zahn, James A; Boyd, Jeffrey M; de la Mora, Arlene M; DiSpirito, Alan A

    2003-10-01

    Improvements in purification of membrane-associated methane monooxygenase (pMMO) have resulted in preparations of pMMO with activities more representative of physiological rates: i.e., >130 nmol.min(-1).mg of protein(-1). Altered culture and assay conditions, optimization of the detergent/protein ratio, and simplification of the purification procedure were responsible for the higher-activity preparations. Changes in the culture conditions focused on the rate of copper addition. To document the physiological events that occur during copper addition, cultures were initiated in medium with cells expressing soluble methane monooxygenase (sMMO) and then monitored for morphological changes, copper acquisition, fatty acid concentration, and pMMO and sMMO expression as the amended copper concentration was increased from 0 (approximately 0.3 microM) to 95 microM. The results demonstrate that copper not only regulates the metabolic switch between the two methane monooxygenases but also regulates the level of expression of the pMMO and the development of internal membranes. With respect to stabilization of cell-free pMMO activity, the highest cell-free pMMO activity was observed when copper addition exceeded maximal pMMO expression. Optimization of detergent/protein ratios and simplification of the purification procedure also contributed to the higher activity levels in purified pMMO preparations. Finally, the addition of the type 2 NADH:quinone oxidoreductase complex (NADH dehydrogenase [NDH]) from M. capsulatus Bath, along with NADH and duroquinol, to enzyme assays increased the activity of purified preparations. The NDH and NADH were added to maintain a high duroquinol/duroquinone ratio. PMID:13129946

  10. Probing the hydrophobic pocket of the active site in the particulate methane monooxygenase (pMMO) from Methylococcus capsulatus (Bath) by variable stereoselective alkane hydroxylation and olefin epoxidation.

    PubMed

    Ng, Kok-Yaoh; Tu, Li-Chun; Wang, Yane-Shih; Chan, Sunney I; Yu, Steve S-F

    2008-05-01

    pMMO from M. capsulatus (Bath) oxidizes straight-chain C1-C5 alkanes and alkenes to form their corresponding 2-alcohols and epoxides. According to experiments performed with cryptically chiral ethane and D,L-[2-(2)H(1),3-(2)H(1)]butane, the reactions proceed through the concerted O-atom insertion mechanism. However, when propene and but-1-ene are used as epoxidation substrates, the enantiomeric excesses (ees) of the enzymatic products are only 18 and 37 %, respectively. This relatively poor stereoselectivity in the enzymatic epoxidation presumably reflects low stereochemical differentiation between the re and si faces in the hydrophobic pocket of the active site. Further insights into the reaction mechanism are now provided by studies on trans-but-2-ene, which reveal only the D,L-2,3-dimethyloxirane products, and on cis-but-2-ene, which yield only the meso product. These observations indicate that the enzymatic epoxidation indeed proceeds through electrophilic syn addition. To achieve better facial selectivity, we have also used 3,3,3-trifluoroprop-1-ene as the substrate. The products obtained are 90 % (2S)-oxirane. When 1,1,1-trifluoropropane is the substrate, the hydroxylation at the 2-carbon exhibits an inverse chiral selectivity relative to that seen with normal butane, if we consider the size of the CF(3) group in the fluorinated propane to be comparable to one of the ethyl groups in butane. These experiments are beginning to delineate the factors that influence the orientations of various substrates in the hydrophobic cavity of the active site in the enzyme. PMID:18383583

  11. Preparation and X-ray structures of metal-free, dicobalt and dimanganese forms of soluble methane monooxygenase hydroxylase from Methylococcus capsulatus (Bath).

    PubMed

    Sazinsky, Matthew H; Merkx, Maarten; Cadieux, Elisabeth; Tang, Sonya; Lippard, Stephen J

    2004-12-28

    A three-component soluble methane monooxygenase (sMMO) enzyme system catalyzes the hydroxylation of methane to methanol at a carboxylate-bridged diiron center housed in the alpha-subunit of the hydroxylase (MMOH). Catalysis is facilitated by the presence of a regulatory protein (MMOB) and inhibited by MMOD, a protein of unknown function encoded in the sMMO operon. Both MMOB and MMOD are presumed to bind to the same region of the MMOH alpha-subunit. A colorimetric method for monitoring removal of Fe(II) from MMOH was developed using 1,10-phenanthroline and yields apo MMOH with <0.1 Fe/homodimer. With the use of this method, it was possible to investigate the X-ray structure of the apoenzyme and to perform metal reconstitution studies. Using MMOH from Methylococccus capsulatus (Bath), the effects of MMOB and MMOD on metal binding were studied and structural perturbations relevant to the function of this enzyme were identified. X-ray crystal structures of the apo, Mn(II)-soaked, and Co(II)-grown MMOH, determined to 2.3 A or greater resolution, reveal that the presence of metal ions is essential for the proper folding of helices E, F, and H of the alpha-subunit. The active sites of Mn(II)-soaked and Co(II)-grown MMOH are similar to that of reduced, native MMOH with notable differences in the metal-metal distances and ligand coordination sphere that may reflect how this dinuclear metal center might change in the presence of MMOB. MMOB and MMOD decrease the rate of removal of Fe(II) from the enzyme by 22- and 16-fold, respectively. On the basis of previous studies, it is hypothesized that MMOB, and perhaps MMOD, function to block solvent access to the MMOH active site. Finally, ITC studies and the observed disorder in helices E, F, and H in the apo and Mn(II)-soaked structures suggest that these regions of MMOH are critical for MMOB and MMOD binding. PMID:15610020

  12. Optimization of solubilization and purification procedures for the hydroxylase component of membrane-bound methane monooxygenase from Methylococcus capsulatus strain M.

    PubMed

    Vasil'ev, V I; Tikhonova, T V; Gvozdev, R I; Tukhvatullin, I A; Popov, V O

    2006-12-01

    The hydroxylase component of membrane-bound (particulate) methane monooxygenase (pMMO) from Methylococcus capsulatus strain M was isolated and purified to homogeneity. The pMMO molecule comprises three subunits of molecular masses 47, 26, and 23 kD and contains three copper atoms and one iron atom. In solution the protein exists as a stable oligomer of 660 kD with possible subunit composition (alpha beta gamma)6. Mass spectroscopy shows high homology of the purified protein with methane monooxygenase from Methylococcus capsulatus strain Bath. Pilot screening of crystallization conditions has been carried out. PMID:17223785

  13. Incomplete tricarboxylic acid cycle in a type I methylotroph, Methylococcus capsulatus.

    PubMed Central

    Patel, R; Hoare, L; Hoare, D S; Taylor, B F

    1975-01-01

    Alpha-Ketoglutaratedehydrogenase was undetectable in extracts of Methylococcus capsulatus. Cells incorporated [1-14-C] acetate into only four protein amino acids (glutamate, proline, arginine, and leucine) and the C5, but not C1, of glutamate. PMID:806581

  14. The C-terminal aqueous-exposed domain of the 45 kDa subunit of the particulate methane monooxygenase in Methylococcus capsulatus (Bath) is a Cu(I) sponge.

    PubMed

    Yu, Steve S-F; Ji, Cheng-Zhi; Wu, Ya Ping; Lee, Tsu-Lin; Lai, Chien-Hung; Lin, Su-Ching; Yang, Zong-Lin; Wang, Vincent C-C; Chen, Kelvin H-C; Chan, Sunney I

    2007-12-01

    The crystal structure of the particulate methane monooxygenase (pMMO) from Methylococcus capsulatus (Bath) has been reported recently [Lieberman, R. L., and Rosenzweig, A. C. (2005) Crystal structure of a membrane-bound metalloenzyme that catalyses the biological oxidation of methane, Nature 434, 177-182]. Subsequent work has shown that the preparation on which the X-ray analysis is based might be missing many of the important metal cofactors, including the putative trinuclear copper cluster at the active site as well as ca. 10 copper ions (E-clusters) that have been proposed to serve as a buffer of reducing equivalents to re-reduce the copper atoms at the active site following the catalytic chemistry [Chan, S. I., Wang, V. C.-C., Lai, J. C.-H., Yu, S. S.-F., Chen, P. P.-Y., Chen, K. H.-C., Chen, C.-L., and Chan, M. K. (2007) Redox potentiometry studies of particulate methane monooxygenase: Support for a trinuclear copper cluster active site, Angew. Chem., Int. Ed. 46, 1992-1994]. Since the aqueous-exposed domains of the 45 kDa subunit (PmoB) have been suggested to be the putative binding domains for the E-cluster copper ions, we have cloned and overexpressed in Escherichia coli the two aqueous-exposed subdomains toward the N- and C-termini of the subunit: the N-terminal subdomain (residues 54-178) and the C-terminal subdomain (residues 257-394 and 282-414). The recombinant C-terminal water-exposed subdomain is shown to behave like a Cu(I) sponge, taking up to ca. 10 Cu(I) ions cooperatively when cupric ions are added to the protein fragment in the presence of dithiothreitol or ascorbate. In addition, circular dichroism measurements reveal that the C-terminal subdomain folds into a beta-sheet structure in the presence of Cu(I). The propensity for the C-terminal subdomain to bind Cu(I) is consistent with the high redox potential(s) determined for the E-cluster copper ions in the pMMO. These properties of the E-clusters are in accordance with the function proposed

  15. Inhibition of Methane Oxidation by Methylococcus capsulatus with Hydrochlorofluorocarbons and Fluorinated Methanes.

    PubMed

    Matheson, L J; Jahnke, L L; Oremland, R S

    1997-07-01

    The inhibition of methane oxidation by cell suspensions of Methylococcus capsulatus (Bath) exposed to hydrochlorofluorocarbon 21 (HCFC-21; difluorochloromethane [CHF(inf2)Cl]), HCFC-22 (fluorodichloromethane [CHFCl(inf2)]), and various fluorinated methanes was investigated. HCFC-21 inhibited methane oxidation to a greater extent than HCFC-22, for both the particulate and soluble methane monooxygenases. Among the fluorinated methanes, both methyl fluoride (CH(inf3)F) and difluoromethane (CH(inf2)F(inf2)) were inhibitory while fluoroform (CHF(inf3)) and carbon tetrafluoride (CF(inf4)) were not. The inhibition of methane oxidation by HCFC-21 and HCFC-22 was irreversible, while that by methyl fluoride was reversible. The HCFCs also proved inhibitory to methanol dehydrogenase, which suggests that they disrupt other aspects of C(inf1) catabolism in addition to methane monooxygenase activity. PMID:16535662

  16. Inhibition of Methane Oxidation by Methylococcus capsulatus with Hydrochlorofluorocarbons and Fluorinated Methanes

    PubMed Central

    Matheson, L. J.; Jahnke, L. L.; Oremland, R. S.

    1997-01-01

    The inhibition of methane oxidation by cell suspensions of Methylococcus capsulatus (Bath) exposed to hydrochlorofluorocarbon 21 (HCFC-21; difluorochloromethane [CHF(inf2)Cl]), HCFC-22 (fluorodichloromethane [CHFCl(inf2)]), and various fluorinated methanes was investigated. HCFC-21 inhibited methane oxidation to a greater extent than HCFC-22, for both the particulate and soluble methane monooxygenases. Among the fluorinated methanes, both methyl fluoride (CH(inf3)F) and difluoromethane (CH(inf2)F(inf2)) were inhibitory while fluoroform (CHF(inf3)) and carbon tetrafluoride (CF(inf4)) were not. The inhibition of methane oxidation by HCFC-21 and HCFC-22 was irreversible, while that by methyl fluoride was reversible. The HCFCs also proved inhibitory to methanol dehydrogenase, which suggests that they disrupt other aspects of C(inf1) catabolism in addition to methane monooxygenase activity. PMID:16535662

  17. The amino acid sequence of cytochrome c-555 from the methane-oxidizing bacterium Methylococcus capsulatus.

    PubMed Central

    Ambler, R P; Dalton, H; Meyer, T E; Bartsch, R G; Kamen, M D

    1986-01-01

    The amino acid sequence of the cytochrome c-555 from the obligate methanotroph Methylococcus capsulatus strain Bath (N.C.I.B. 11132) was determined. It is a single polypeptide chain of 96 residues, binding a haem group through the cysteine residues at positions 19 and 22, and the only methionine residue is a position 59. The sequence does not closely resemble that of any other cytochrome c that has yet been characterized. Detailed evidence for the amino acid sequence of the protein has been deposited as Supplementary Publication SUP 50131 (12 pages) at the British Library Lending Division, Boston Spa, West Yorkshire LS23 7BQ, U.K., from whom copies are available on prepayment. PMID:3006666

  18. Inhibition of methane oxidation by Methylococcus capsulatus with hydrochlorofluorocarbons and fluorinated methanes

    USGS Publications Warehouse

    Matheson, L.J.; Jahnke, L.L.; Oremland, R.S.

    1997-01-01

    The inhibition of methane oxidation by cell suspensions of Methylococcus capsulatus (Bath) exposed to hydrochlorofluorocarbon 21 (HCFC-21; difluorochloromethane [CHF2Cl]), HCFC-22 (fluorodichloromethane [CHFCl2]), and various fluorinated methanes was investigated. HCFC-21 inhibited methane oxidation to a greater extent than HCFC-22, for both the particulate and soluble methane monooxygenases. Among the fluorinated methanes, both methyl fluoride (CH3F) and difluoromethane (CH2F2) were inhibitory while fluoroform (CHF3) and carbon tetrafluoride (CF4) were not. The inhibition of methane oxidation by HCFC-21 and HCFC-22 was irreversible, while that by methyl fluoride was reversible. The HCFCs also proved inhibitory to methanol dehydrogenase, which suggests that they disrupt other aspects of C1 catabolism in addition to methane monooxygenase activity.

  19. Copper-dependent reciprocal transcriptional regulation of methane monooxygenase genes in Methylococcus capsulatus and Methylosinus trichosporium.

    PubMed

    Nielsen, A K; Gerdes, K; Murrell, J C

    1997-07-01

    The methanotrophic bacteria Methylococcus capsulatus (Bath) and Methylosinus trichosporium OB3b convert methane to methanol using the enzyme, methane monooxygenase (MMO). These bacteria are able to express two distinct MMOs: a cytoplasmic or soluble form (sMMO) and a membrane-bound or particulate form (pMMO). Differential expression of sMMO and pMMO is regulated by the amount of copper ions available to the cells; sMMO is expressed at low copper-biomass ratios, whereas pMMO is expressed at high copper-biomass ratios. In both methanotrophs, transcription of the sMMO gene cluster is negatively regulated by copper ions. Data suggest that transcription of the M. trichosporium OB3b sMMO gene cluster is directed from a sigma54-like and a sigma70-like promoter. The pMMO (pmo) genes of M. capsulatus (Bath) are transcribed into a polycistronic mRNA of 3.3 kb. The synthesis of this mRNA was activated by copper ions. Activation of pmo transcription by copper ions was concomitant with repression of sMMO gene transcription in both methanotrophs. This suggests that a common regulatory pathway may be involved in the transcriptional switch between sMMO and pMMO gene expression. PMID:9282751

  20. Synthesis of cell constituents by methane-grown Methylococcus capsulatus and Methanomonas methanooxidans

    PubMed Central

    Lawrence, A. J.; Kemp, M. B.; Quayle, J. R.

    1970-01-01

    1. A study was made of the incorporation of carbon from [14C]methanol by cultures of Methylococcus capsulatus and Methanomonas methanooxidans growing on methane. 2. The distribution of radioactivity within the non-volatile constituents of the ethanol-soluble fractions of the cells, after incubation with labelled substrate for periods of up to 3min, was analysed by chromatography and radioautography. 3. Over 80% of the radioactivity fixed by Methylococcus capsulatus at 30°C at the earliest times of sampling appeared in phosphorylated compounds, of which glucose phosphate constituted 60%. 4. Most of the radioactivity fixed by Methanomonas methanooxidans at 30°C at the earliest times of sampling appeared in serine, malate, aspartate and an unknown compound(s) tentatively suggested to be folate derivative(s). At 16°C, [14C]methanol was fixed predominantly into serine and the unknown compound(s). 5. Extracts of Methylococcus capsulatus contain an enzyme system that catalyses the condensation of formaldehyde and ribose 5-phosphate to give a mixture consisting mainly of fructose phosphate and allulose phosphate. No similar activity was detected in extracts of Methanomonas methanooxidans. A convenient method was developed for assay of this enzyme system. 6. The enzyme system catalysing the condensation of formaldehyde with ribose 5-phosphate is particle-bound in both Methylococcus capsulatus and Pseudomonas methanica and is unstable in the absence of Mg2+. 7. Extracts of Methanomonas methanooxidans contain high activities of d-glycerate–NAD oxidoreductase, whereas extracts of Methylococcus capsulatus and Pseudomonas methanica contain negligible activities of this enzyme. 8. These results indicate that during growth of Methylococcus capsulatus on methane, as with Pseudomonas methanica, cell constituents are made by the ribose phosphate cycle of formaldehyde fixation. This contrasts with Methanomonas methanooxidans, whose assimilation pathway resembles in some features

  1. An oxidized tryptophan facilitates copper binding in Methylococcus capsulatus-secreted protein MopE.

    PubMed

    Helland, Ronny; Fjellbirkeland, Anne; Karlsen, Odd Andre; Ve, Thomas; Lillehaug, Johan R; Jensen, Harald B

    2008-05-16

    Proteins can coordinate metal ions with endogenous nitrogen and oxygen ligands through backbone amino and carbonyl groups, but the amino acid side chains coordinating metals do not include tryptophan. Here we show for the first time the involvement of the tryptophan metabolite kynurenine in a protein metal-binding site. The crystal structure to 1.35 angstroms of MopE* from the methane-oxidizing Methylococcus capsulatus (Bath) provided detailed information about its structure and mononuclear copper-binding site. MopE* contains a novel protein fold of which only one-third of the structure displays similarities to other known folds. The geometry around the copper ion is distorted tetrahedral with one oxygen ligand from a water molecule, two histidine imidazoles (His-132 and His-203), and at the fourth distorted tetrahedral position, the N1 atom of the kynurenine, an oxidation product of Trp-130. Trp-130 was not oxidized to kynurenine in MopE* heterologously expressed in Escherichia coli, nor did this protein bind copper. Our findings indicate that the modification of tryptophan to kynurenine and its involvement in copper binding is an innate property of M. capsulatus MopE*. PMID:18348978

  2. Draft Genome Sequence of the Methane-Oxidizing Bacterium Methylococcus capsulatus (Texas)

    PubMed Central

    Hult, Lene T. Olsen; Kuczkowska, Katarzyna; Jacobsen, Morten; Lea, Tor; Pope, Phillip B.

    2012-01-01

    Methanotrophic bacteria perform major roles in global carbon cycles via their unique enzymatic activities that enable the oxidation of one-carbon compounds, most notably methane. Here we describe the annotated draft genome sequence of the aerobic methanotroph Methylococcus capsulatus (Texas), a type strain originally isolated from sewer sludge. PMID:23144383

  3. Inhibition of dimethyl ether and methane oxidation in Methylococcus capsulatus and Methylosinus trichosporium.

    PubMed Central

    Patel, R; Hou, C T; Felix, A

    1976-01-01

    Metal-chelating or -binding agents inhibited the oxidation of dimethyl ether and methane, but not methanol, by cell suspensions of Methylococcus capsulatus and Methylosinus trichosporium. Evidence suggests that the involvement of metal-containing enzymatic systems in the initial step of oxidation of dimethyl ether and methane. PMID:4428

  4. Draft genome sequence of the methane-oxidizing bacterium Methylococcus capsulatus (Texas).

    PubMed

    Kleiveland, Charlotte R; Hult, Lene T Olsen; Kuczkowska, Katarzyna; Jacobsen, Morten; Lea, Tor; Pope, Phillip B

    2012-12-01

    Methanotrophic bacteria perform major roles in global carbon cycles via their unique enzymatic activities that enable the oxidation of one-carbon compounds, most notably methane. Here we describe the annotated draft genome sequence of the aerobic methanotroph Methylococcus capsulatus (Texas), a type strain originally isolated from sewer sludge. PMID:23144383

  5. High-Molecular-Mass Multi-c-Heme Cytochromes from Methylococcus capsulatus Bath†

    PubMed Central

    Bergmann, David J.; Zahn, James A.; DiSpirito, Alan A.

    1999-01-01

    The polypeptide and structural gene for a high-molecular-mass c-type cytochrome, cytochrome c553O, was isolated from the methanotroph Methylococcus capsulatus Bath. Cytochrome c553O is a homodimer with a subunit molecular mass of 124,350 Da and an isoelectric point of 6.0. The heme c concentration was estimated to be 8.2 ± 0.4 mol of heme c per subunit. The electron paramagnetic resonance spectrum showed the presence of multiple low spin, S = 1/2, hemes. A degenerate oligonucleotide probe synthesized based on the N-terminal amino acid sequence of cytochrome c553O was used to identify a DNA fragment from M. capsulatus Bath that contains occ, the gene encoding cytochrome c553O. occ is part of a gene cluster which contains three other open reading frames (ORFs). ORF1 encodes a putative periplasmic c-type cytochrome with a molecular mass of 118,620 Da that shows approximately 40% amino acid sequence identity with occ and contains nine c-heme-binding motifs. ORF3 encodes a putative periplasmic c-type cytochrome with a molecular mass of 94,000 Da and contains seven c-heme-binding motifs but shows no sequence homology to occ or ORF1. ORF4 encodes a putative 11,100-Da protein. The four ORFs have no apparent similarity to any proteins in the GenBank database. The subunit molecular masses, arrangement and number of hemes, and amino acid sequences demonstrate that cytochrome c553O and the gene products of ORF1 and ORF3 constitute a new class of c-type cytochrome. PMID:9922265

  6. Reduction of RNA and DNA in Methylococcus capsulatus by endogenous nucleases.

    PubMed

    Larsen, J; Joergensen, L

    1996-03-01

    A method for reducing RNA and DNA in the bacterium Methylococcus capsulatus (Bath) has been developed. Endogenous RNase and DNase were activated by a 10 s heat shock at 90 degrees C. Cells were then incubated at 60 degrees C for 20 min to allow degradation of the nucleic acids. The optimum pH for the process was 7.0. The protein loss was less than 10% and occurred during the initial heat shock. No protein loss was found during incubation. The total dry-weight loss in connection with an 80% reduction of the nucleic acid content was 20%-25%, giving a final product with a raw protein content of approximately 75%. Reduction of both RNA and DNA was inhibited by CuSO4 and ZnSO4. DNA reduction was stimulated by other minerals. Optimal stimulation was found at 1 mM FeSO4. Reduction of RNA was not increased by any of the minerals tested. PMID:8920188

  7. (14C)acetate assimilation by a type I obligate methylotroph, Methylococcus capsulatus.

    PubMed Central

    Patel, R N; Hoare, S L; Hoare, D S; Taylor, B F

    1977-01-01

    Methanol and formate oxidation supported the assimilation of [14C]acetate by cell suspensions of Methylococcus capsulatus; oxidation of other primary alcohols, except ethanol, did not. The extent of [1-14C]acetate assimilation supported by methanol oxidation was decreased in the presence of primary alcohols, except ethanol. Potassium cyanide (0.33 mM) completely inhibited the oxidation of formate and its stimulation of [1-14C]acetate assimilation. The amount of [1-14C]acetate assimilation supported by methanol oxidation was significantly inhibited by cyanide. PMID:412469

  8. An EPR study of the dinuclear iron site in the soluble methane monooxygenase from Methylococcus capsulatus (Bath) reduced by one electron at 77 K: the effects of component interactions and the binding of small molecules to the diiron(III) center.

    PubMed

    Davydov, R; Valentine, A M; Komar-Panicucci, S; Hoffman, B M; Lippard, S J

    1999-03-30

    Reduction of the soluble methane monooxygenase hydroxylase (MMOH) from Methylococcus capsulatus (Bath) in frozen 4:1 buffer/glycerol solutions at 77 K by mobile electrons generated by gamma-irradiation produces an EPR-detectable, mixed-valent Fe(II)Fe(III) center. At this temperature the conformation of the enzyme remains essentially unaltered during reduction, so the mixed-valent EPR spectra serve to probe the active site structure of the EPR-silent, diiron(III) state. The EPR spectra of the cryoreduced samples reveal that the diiron(III) cluster of the resting hydroxylase has at least two chemically distinct forms, the structures of which differ from that of the equilibrium Fe(II)Fe(III) site. Their relative populations depend on pH, the presence of component B, and formation of the MMOH/MMOB complex by reoxidation of the reduced, diiron(II) hydroxylase. The formation of complexes between MMOB, MMOR, and the oxidized hydroxylase does not measurably affect the structure of the diiron(III) site. Cryogenic reduction in combination with EPR spectroscopy has also provided information about interaction of MMOH in the diiron(III) state with small molecules. The diiron(III) center binds methanol and phenols, whereas DMSO and methane have no measurable effect on the EPR properties of cryoreduced hydroxylase. Addition of component B favors the binding of some exogenous ligands, such as DMSO and glycerol, to the active site diiron(III) state and markedly perturbs the structure of the diiron(III) cluster complexed with methanol or phenol. The results reveal different reactivity of the Fe(III)Fe(III) and Fe(II)Fe(III) redox states of MMOH toward exogenous ligands. Moreover, unlike oxidized hydroxylase, the binding of exogenous ligands to the protein in the mixed-valent state is allosterically inhibited by MMOB. The differential reactivity of the hydroxylase in its diiron(III) and mixed-valent states toward small molecules, as well as the structural basis for the regulatory

  9. Evidence that a type-2 NADH:quinone oxidoreductase mediates electron transfer to particulate methane monooxygenase in methylococcus capsulatus.

    PubMed

    Cook, Scott A; Shiemke, Andrew K

    2002-02-01

    NADH readily provides reducing equivalents to membrane-bound methane monooxygenase (pMMO) from Methylococcus capsulatus (Bath) in isolated membrane fractions, but detergent solubilization disrupts this electron-transfer process. Addition of exogenous quinones (especially decyl-plastoquinone and duroquinone) restores the NADH-dependent pMMO activity. Results of inhibitor and substrate dependence of this activity indicate the presence of only a type-2 NADH:quinone oxidoreductase (NDH-2). A 100-fold purification of the NDH-2 was achieved using lauryl-maltoside solubilization followed by ion exchange, hydrophobic-interaction, and gel-filtration chromatography. The purified NDH-2 has a subunit molecular weight of 36 kDa and exists as a monomer in solution. UV-visible and fluorescence spectroscopy identified flavin adenine dinucleotide (FAD) as a cofactor present in stoichiometric amounts. NADH served as the source of electrons, whereas NADPH could not. The purified NDH-2 enzyme reduced coenzyme Q(0), duroquinone, and menaquinone at high rates, whereas the decyl analogs of ubiquinone and plastoquinone were reduced at approximately 100-fold lower rates. Rotenone and flavone did not inhibit the NDH-2, whereas amytal caused partial inhibition but only at high concentrations. PMID:11811946

  10. Hexose phosphate synthetase from Methylococcus capsulatus makes d-arabino-3-hexulose phosphate

    PubMed Central

    Kemp, M. B.

    1974-01-01

    The product of the reaction catalysed by hexose phosphate synthase prepared from Methylococcus capsulatus was dephosphorylated and the sugar moiety purified. The sugar and derivatives were compared by various chromatographic and other methods with authentic samples of allulose (psicose), d-erythro-l-glycero-3-hexulose and d-erythro-d-glycero-3-hexulose. The sugar is not allulose, as was previously thought on the basis of less extensive evidence (Kemp & Quayle, 1966), but is in fact d-erythro-l-glycero-3-hexulose (d-arabino-3-hexulose). This identification is consistent with recent studies which have shown that hexose phosphate synthase catalyses the condensation of formaldehyde with d-ribulose 5-phosphate rather than with d-ribose 5-phosphate (Kemp, 1972). PMID:4463938

  11. Lanosterol biosynthesis in the prokaryote Methylococcus capsulatus: insight into the evolution of sterol biosynthesis.

    PubMed

    Lamb, David C; Jackson, Colin J; Warrilow, Andrew G S; Manning, Nigel J; Kelly, Diane E; Kelly, Steven L

    2007-08-01

    A putative operon containing homologues of essential eukaryotic sterol biosynthetic enzymes, squalene monooxygenase and oxidosqualene cyclase, has been identified in the genome of the prokaryote Methylococcus capsulatus. Expression of the squalene monooxygenase yielded a protein associated with the membrane fraction, while expression of oxidosqualene cyclase yielded a soluble protein, contrasting with the eukaryotic enzyme forms. Activity studies with purified squalene monooxygenase revealed a catalytic activity in epoxidation of 0.35 nmol oxidosqualene produced/min/nmol squalene monooxygenase, while oxidosqualene cyclase catalytic activity revealed cyclization of oxidosqualene to lanosterol with 0.6 nmol lanosterol produced/min/nmol oxidosqualene cyclase and no other products observed. The presence of prokaryotic sterol biosynthesis is still regarded as rare, and these are the first representatives of such prokaryotic enzymes to be studied, providing new insight into the evolution of sterol biosynthesis in general. PMID:17567593

  12. Methyl sterol and cyclopropane fatty acid composition of Methylococcus capsulatus grown at low oxygen tensions.

    PubMed Central

    Jahnke, L L; Nichols, P D

    1986-01-01

    Methylococcus capsulatus contained extensive intracytoplasmic membranes when grown in fed-batch cultures over a wide range of oxygen tensions (0.1 to 10.6%, vol/vol) and at a constant methane level. Although the biomass decreased as oxygen levels were lowered, consistently high amounts of phospholipid and methyl sterol were synthesized. The greatest amounts of sterol and phospholipid were found in cells grown between 0.5 and 1.1% oxygen (7.2 and 203 mumol/g [dry weight], respectively). While sterol was still synthesized in significant amounts in cells grown at 0.1% oxygen, the major sterol product was the dimethyl form. Analysis by capillary gas chromatography-mass spectrophotometry showed that the phospholipid esterified fatty acids were predominantly 16:0 and 16:1 and that the hexadecenoates consisted of cis delta 9, delta 10, and delta 11 isomers. At low oxygen tensions, the presence of large amounts (25%) of cyclopropane fatty acids (cy 17:0) with the methylene groups at the delta 9, delta 10, and delta 11 positions was detected. Although the delta 9 monoenoic isomer was predominant, growth at low oxygen levels enhanced the synthesis of the delta 10 isomers of 16:1 and cy 17:0. As the oxygen level was increased, the amount of cyclopropanes decreased, such that only a trace of cy 17:0 could be detected in cells grown at 10.6% oxygen. Although M. capsulatus grew at very low oxygen tensions, this growth was accompanied by changes in the membrane lipids. PMID:3087955

  13. Hydrogen isotope fractionation in lipids of the methane-oxidizing bacterium Methylococcus capsulatus

    NASA Astrophysics Data System (ADS)

    Sessions, Alex L.; Jahnke, Linda L.; Schimmelmann, Arndt; Hayes, John M.

    2002-11-01

    Hydrogen isotopic compositions of individual lipids from Methylococcus capsulatus, an aerobic, methane-oxidizing bacterium, were analyzed by hydrogen isotope-ratio-monitoring gas chromatography-mass spectrometry (GC-MS). The purposes of the study were to measure isotopic fractionation factors between methane, water, and lipids and to examine the biochemical processes that determine the hydrogen isotopic composition of lipids. M. capsulatus was grown in six replicate cultures in which the δD values of methane and water were varied independently. Measurement of concomitant changes in δD values of lipids allowed estimation of the proportion of hydrogen derived from each source and the isotopic fractionation associated with the utilization of each source. All lipids examined, including fatty acids, sterols, and hopanols, derived 31.4 ± 1.7% of their hydrogen from methane. This was apparently true whether the cultures were harvested during exponential or stationary phase. Examination of the relevant biochemical pathways indicates that no hydrogen is transferred directly (with C-H bonds intact) from methane to lipids. Accordingly, we hypothesize that all methane H is oxidized to H 2O, which then serves as the H source for all biosynthesis, and that a balance between diffusion of oxygen and water across cell membranes controls the concentration of methane-derived H 2O at 31%. Values for α l/ w, the isotopic fractionation between lipids and water, were 0.95 for fatty acids and 0.85 for isoprenoid lipids. These fractionations are significantly smaller than those measured in higher plants and algae. Values for α l/ m, the isotopic fractionation between lipids and methane, were 0.94 for fatty acids and 0.79 for isoprenoid lipids. Based on these results, we predict that methanotrophs living in seawater and consuming methane with typical δD values will produce fatty acids with δD between -50 and -170‰, and sterols and hopanols with δD between -150 and -270‰.

  14. Biochemical characterization of isocitrate dehydrogenase from Methylococcus capsulatus reveals a unique NAD+-dependent homotetrameric enzyme.

    PubMed

    Stokke, Runar; Madern, Dominique; Fedøy, Anita-Elin; Karlsen, Solveig; Birkeland, Nils-Kåre; Steen, Ida Helene

    2007-05-01

    The gene encoding isocitrate dehydrogenase (IDH) of Methylococcus capsulatus (McIDH) was cloned and overexpressed in Escherichia coli. The purified enzyme was NAD+-dependent with a thermal optimum for activity at 55-60 degrees C and an apparent midpoint melting temperature (Tm) of 70 degrees C. Analytical ultracentrifugation (AUC) revealed a homotetrameric state, and McIDH thus represents the first homotetrameric NAD+-dependent IDH that has been characterized. Based on a structural alignment of McIDH and homotetrameric homoisocitrate dehydrogenase (HDH) from Thermus thermophilus (TtHDH), we identified the clasp-like domain of McIDH as a likely site for tetramerization. McIDH showed moreover, higher sequence identity (48%) to TtHDH than to previously characterized IDHs. Putative NAD+-IDHs with high sequence identity (48-57%) to McIDH were however identified in a variety of bacteria showing that NAD+-dependent IDHs are indeed widespread within the domain, Bacteria. Phylogenetic analysis including these new sequences revealed a close relationship with eukaryal allosterically regulated NAD+-IDH and the subfamily III of IDH was redefined to include bacterial NAD+- and NADP+-dependent IDHs. This apparent relationship suggests that the mitochondrial genes encoding NAD+-IDH are derived from the McIDH-like IDHs. PMID:17160675

  15. [Purification and properties of membrane-bound methane hydroxylase from Methylococcus capsulatus (strain M)].

    PubMed

    Gvozdev, R I; Tukhvatullin, I A; Tumanova, L V

    2008-01-01

    Membrane fraction of Methylococcus capsulatus (strain M) were treated with [14C]acetylene, an affinity label binding to the active center of membrane-bound methane monooxygenase (MMO). High-purity particulate form of methane hydroxylase (pMH) was obtained by ion exchange and hydrophobic chromatography. According to SDS-PAGE data, the enzyme contained three polypeptides with molecular weights of 47 (alpha), 27 (beta), and 25 (gamma) kDa in the ratio 1:1:1. The radiolabel was contained in the beta-subunit of pMH. The protein contained 1 or 2 atoms of nonheme iron and 2-4 atoms of copper per a minimum molecular weight of 99 kDa. This protein did not oxidize methane or propylene in the presence of NADH but was able to oxidize low quantities of methane in the presence of duroquinol. It was established that methanol dehydrogenase (MD) and NADH oxidoreductase (NADH-OR) are peripheral membrane proteins. Possible causes of low activity of high-purity methane hydroxylase are discussed. PMID:18946992

  16. Physiological Studies of Methane- and Methanol-Oxidizing Bacteria: Immunological Comparison of a Primary Alcohol Dehydrogenase from Methylococcus capsulatus and Pseudomonas sp. M27

    PubMed Central

    Patel, R. N.; Mandy, W. J.; Hoare, D. S.

    1973-01-01

    A primary alcohol dehydrogenase was purified from cell extracts of two apparently unrelated microorganisms, namely, Pseudomonas sp. M27 and Methylococcus capsulatus. Rabbit antiserum prepared against the purified enzyme from M. capsulatus revealed distinctive antigenic determinants by quantitative and gel precipitin reactions. Rabbit antiserum to M27 enzyme detected both distinctive and shared antigenic determinants. Certain methane- and methanol-oxidizing bacteria were grouped on the basis of serological cross-reacting enzyme specificities. Images PMID:4120569

  17. Pteridines produced by Methylococcus capsulatus. Isolation and identification of a neopterin 2′:3′-phosphate

    PubMed Central

    Urushibara, T.; Forrest, H. S.; Hoare, D. S.; Patel, R. N.

    1971-01-01

    Three pteridines have been isolated from the methane- or methanol-oxidizing bacterium Methylococcus capsulatus. Two of these are known compounds, 2-amino-6-carboxy-4-hydroxypteridine and 2-amino-4-hydroxy-6-methylpteridine. The third is shown by degradative and synthetic experiments to be l-threo-neopterin 2′:3′-phosphate. Labelling experiments show that both the pteridine moiety and phosphate residue are derived from a single GTP molecule. The possible metabolic significance of these compounds in methanol oxidation is discussed. PMID:5158900

  18. Purification and properties of 3-hexulose phosphate synthase and phospho-3-hexuloisomerase from Methylococcus capsulatus

    PubMed Central

    Ferenci, Thomas; Strøm, Terje; Quayle, J. Rodney

    1974-01-01

    3-Hexulose phosphate synthase and phospho-3-hexuloisomerase were purified 40- and 150-fold respectively from methane-grown Methylococcus capsulatus. The molecular weights of the enzymes were approximately 310000 and 67000 respectively, as determined by gel filtration. Dissociation of 3-hexulose phosphate synthase into subunits of molecular weight approx. 49000 under conditions of low pH or low ionic strength was observed. Within the range of compounds tested, 3-hexulose phosphate synthase is specific for formaldehyde and d-ribulose 5-phosphate (forward reaction) and d-arabino-3-hexulose 6-phosphate (reverse reaction), and phospho-3-hexuloisomerase is specific for d-arabino-3-hexulose 6-phosphate (forward reaction) and d-fructose 6-phosphate (reverse reaction). A bivalent cation is essential for activity and stability of 3-hexulose phosphate synthase; phospho-3-hexuloisomerase is inhibited by many bivalent cations. The pH optima of the two enzymes are 7.0 and 8.3 respectively and the equilibrium constants are 4.0×10−5m and 1.9×102m respectively. The apparent Michaelis constants for 3-hexulose phosphate synthase are: d-ribulose 5-phosphate, 8.3×10−5m; formaldehyde, 4.9×10−4m; d-arabino-3-hexulose 6-phosphate, 7.5×10−5m. The apparent Michaelis constants for phospho-3-hexuloisomerase are: d-arabino-3-hexulose 6-phosphate, 1.0×10−4m; d-fructose 6-phosphate, 1.1×10−3m. PMID:4219834

  19. [The binuclear iron site of the membrane-bound methane hydroxylase from Methylococcus capsulatus (strain M)].

    PubMed

    Tumanova, L V; Tukhvatullin, I A; Burbaev, T Sh; Gvozdev, R I; Andersson, K K

    2008-01-01

    The particulate membrane-bound methane hydroxylase (pMMOH) was isolated from methane-oxidizing cells of Methylococcus capsulatus (strain M). At SDS PAGE, pMMOH displays three bands: 47 (alpha), 27 (beta), and 25 kDa (gamma). The ESR spectrum of pMMOH incubated with hydrogen peroxide (final concentration 20 mM) at 4 degrees C exhibited, along with the copper signal of type I with g = 2.05, signals of cytochrome with g = 3.0 and of high-spin ferriheme with g = 6.00. After incubation at -30 degrees C, additional signals with g 8.5 and 13.5 were observed. These signals, which have not been recorded previously in pMMOH preparations, are due to an intermediate of the pMMOH active site, which arises in the reaction of hydrogen peroxide with pMMOH at -30 degrees C. It was established that this intermediate is a high-spin dimer [Fe(IlI)-Fe(IV)] with S = 9/2 and different degree of rhombic distortion of structure (it is responsible for both signals). Presumably, the signal with g = 8.5 also arises from the same dimer [Fe(III)-Fe(IV)], but with S = 7/2. The presence of the intermediate [Fe(lII)-Fe(IV)] in pMMOH preparations suggests that the original state of the pMMOH active site is the dimer [Fe(III)-Fe(III)] which is located in the beta-subunit and cannot be detected by ESR. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2008, vol. 34, no. 2; see also http:// www.maik.ru. PMID:18522275

  20. Physiological Studies of Methane and Methanol-Oxidizing Bacteria: Oxidation of C-1 Compounds by Methylococcus capsulatus

    PubMed Central

    Patel, Ramesh N.; Hoare, Derek S.

    1971-01-01

    Methylococcus capsulatus grows only on methane or methanol as its sole source of carbon and energy. Some amino acids serve as nitrogen sources and are converted to keto acids which accumulate in the culture medium. Cell suspensions oxidize methane, methanol, formaldehyde, and formate to carbon dioxide. Other primary alcohols are oxidized only to the corresponding aldehydes. Oxidation of formate by cell suspensions is more sensitive to inhibition by cyanide than is the oxidation of other one carbon compounds. This is due to the cyanide sensitivity of a soluble nicotinamide adenine dinucleotide-specific formate dehydrogenase. Oxidation of formaldehyde and methanol is catalyzed by a nonspecific primary alcohol dehydrogenase which is activated by ammonium ions and is independent of pyridine nucleotides. Some comparisons are made with a strain of Pseudomonas methanica. PMID:5563868

  1. Oxidation of C1 compounds by particulate fractions from Methylococcus capsulatus: properties of methanol oxidase and methanol dehydrogenase.

    PubMed Central

    Wadzinski, A M; Ribbons, D W

    1975-01-01

    Methanol (and formaldehyde) oxidizing activities in crude extracts of Methylococcus capsulatus are associated mainly with particulate fractions sedimenting between 3,000 and 40,000 X g. Most of the phenazine methosulfate (PMS)-dependent methanol (and formaldehyde) dehydrogenase activity observed resides in the soluble fraction but represents only 40% of the total (PMS dependent plus independent) activity. Both PMS-dependent methanol dehydrogenase activity and PMS-independent methanol oxidase activity are found in particulate fractions, and the PMS-dependent dehydrogenase is easily solubilized by treatment with certain phospholipases or detergents. The properties of the PMS-dependent dehydrogenase activities in the soluble fraction and that solubilized from the particles suggested that they may be identical proteins. Their pH optima, temperature dependence, thermolabilities, and sensitivities to the presence of specific antisera were indistinguishable. Homogeneous preparations of the enzyme proteins obtained from the soluble fractions of extracts and the particulate fractions solubilized by detergents had similar: (i) electrophoretic mobilities in native and denatured states (subunit size in sodium dodecyl sulfate 62,000 daltons); (ii) molecular radii under native conditions, (iii) visible absorption spectra, lambdamax 350 nm, (iv) kinetic constants for methanol and formaldehyde; (v) substrate specificity; and (vi) immunological characteristics--antisera to each enzyme preparation showed precipitin lines of identity to either of the enzymes. It is suggested that the major site of methanol and formaldehyde oxidation in M. capsulatus occurs on the intracytoplasmic membranes in vivo and is coupled to oxygen reduction. Images PMID:238947

  2. The carbon assimilation pathways of Methylococcus capsulatus, Pseudomonas methanica and Methylosinus trichosporium (OB3B) during growth on methane

    PubMed Central

    Strøm, Terje; Ferenci, Thomas; Quayle, J. Rodney

    1974-01-01

    d-arabino-3-Hexulose 6-phosphate was prepared by condensation of formaldehyde with ribulose 5-phosphate in the presence of 3-hexulose phosphate synthase from methane-grown Methylococcus capsulatus. The 3-hexulose phosphate was unstable in solutions of pH greater than 3, giving a mixture of products in which, after dephosphorylation, allulose and fructose were detected. A complete conversion of d-ribulose 5-phosphate and formaldehyde into d-fructose 6-phosphate was demonstrated in the presence of 3-hexulose phosphate synthase and phospho-3-hexuloisomerase (prepared from methane-grown M. capsulatus). d-Allulose 6-phosphate was prepared from d-allose by way of d-allose 6-phosphate. No evidence was found for its metabolism by extracts of M. capsulatus, thus eliminating it as an intermediate in the carbon assimilation process of this organism. A survey was made of the enzymes involved in the regeneration of pentose phosphate during C1 assimilation via a modified pentose phosphate cycle. On the basis of the presence of the necessary enzymes, two alternative routes for cleavage of fructose 6-phosphate are suggested, one route involves fructose diphosphate aldolase and the other 6-phospho-2-keto-3-deoxygluconate aldolase. A detailed formulation of the complete ribulose monophosphate cycle of formaldehyde fixation is presented. The energy requirements for carbon assimilation by this cycle are compared with those for the serine pathway and the ribulose diphosphate cycle of carbon dioxide fixation. A cyclic scheme for oxidation of formaldehyde via 6-phosphogluconate is suggested. PMID:4377654

  3. Novel hopanoids from the methylotrophic bacteria Methylococcus capsulatus and Methylomonas methanica. (22S)-35-aminobacteriohopane-30,31,32,33,34-pentol and (22S)-35-amino-3 beta-methylbacteriohopane-30,31,32,33,34-pentol.

    PubMed Central

    Neunlist, S; Rohmer, M

    1985-01-01

    The major hopanoid of the methylotrophic bacteria Methylococcus capsulatus and Methylomonas methanica was identified by spectroscopic methods as (22S)-35-aminobacteriohopane-30,31,32,33,34-pentol. Minor companions were, in both bacteria, 35-aminobacteriohopane-31,32,33,34-tetrol and in Methylomonas methanica, 35-aminobacteriohopane-32,33,34-triol. In Methylococcus capsulatus the aminopentol and the aminotetrol were accompanied by their homologues possessing an extra methyl group at C-3. Bacterial hopanoids with a functionalized C-30 carbon atom such as these two new aminopentols are possible precursors of widespread C29 hopanoid chemical fossils. PMID:3935106

  4. Physiological Studies of Methane- and Methanol-Oxidizing Bacteria: Comparison of a Primary Alcohol Dehydrogenase from Methylococcus capsulatus (Texas Strain) and Pseudomonas Species M27

    PubMed Central

    Patel, R. N.; Bose, H. R.; Mandy, W. J.; Hoare, D. S.

    1972-01-01

    A primary alcohol dehydrogenase has been purified from Methylococcus capsulatus (Texas strain). The purified enzyme catalyzes the oxidation of methanol and formaldehyde to formate; other primary alcohols are oxidized to their corresponding aldehydes. Ammonium ions are required for enzyme activity. The enzyme has a molecular weight of 120,000 daltons and consists of two 62,000 molecular-weight subunits which dissociate at acidic pH. The enzyme is similar to an alcohol dehydrogenase enzyme isolated from Pseudomonas sp. M27. Images PMID:5022170

  5. Methyl sterol and cyclopropane fatty acid composition of Methylococcus capsulatus grown at low oxygen tensions

    NASA Technical Reports Server (NTRS)

    Jahnke, L. L.; Nichols, P. D.

    1986-01-01

    The sterol and fatty acid concentrations for M. capsulatus grown in fed-batch cultures over a wide range of oxygen tensions (0.1-10.6 percent) and at a constant methane level are evaluated. The analyses reveal that the biomass decreases as oxygen levels are lowered; the sterol concentration increases when the oxygen range is between 0.5-1.1 percent and decreases when the oxygen range is below 0.5 percent; and the amount of monounsaturated C16 decreases and the concentration of cyclopropane fatty acids increases after oxygen is reduced. It is noted that growth and membrane synthesis occur at low oxygen concentrations and that the synthesis of membrane lipids responds to growth conditions.

  6. A novel sterol 14alpha-demethylase/ferredoxin fusion protein (MCCYP51FX) from Methylococcus capsulatus represents a new class of the cytochrome P450 superfamily.

    PubMed

    Jackson, Colin J; Lamb, David C; Marczylo, Timothy H; Warrilow, Andrew G S; Manning, Nigel J; Lowe, David J; Kelly, Diane E; Kelly, Steven L

    2002-12-01

    Sterol 14alpha-demethylase encoded by CYP51 is a member of the cytochrome P450 (CYP) superfamily of enzymes and has been shown to have an essential role in sterol biosynthesis in eukaryotes, with orthologues recently being described in some bacteria. Examination of the genome sequence data for the proteobacterium Methylococcus capsulatus, a bacterial species known to produce sterol, revealed the presence of a single CYP with strong homology to CYP51, particularly to a form in Mycobacterium tuberculosis. This M. capsulatus CYP51 protein represents a new class of CYP consisting of the CYP domain naturally fused to a ferredoxin domain at the C terminus via an alanine-rich linker. Expression of the M. capsulatus MCCYP51FX fusion in Escherichia coli yielded a P450, which, when purified to homogeneity, had the predicted molecular mass approximately 62 kDa on SDS/PAGE and bound lanosterol as a putative substrate. Sterol 14alpha-demethylase activity was shown (0.24 nmol of lanosterol metabolized per minute per nanomole of MCCYP51FX fusion) by gas chromatography/mass spectrometry with the activity dependent upon the presence of ferredoxin reductase and NADPH. Our unique findings describe a new class of naturally existing cytochrome P450, which will provide pivotal information for CYP structure/function in general. PMID:12235134

  7. Sterol biosynthesis by a prokaryote: first in vitro identification of the genes encoding squalene epoxidase and lanosterol synthase from Methylococcus capsulatus.

    PubMed

    Nakano, Chiaki; Motegi, Akihiro; Sato, Tsutomu; Onodera, Masayuki; Hoshino, Tsutomu

    2007-10-01

    Sterol biosynthesis by prokaryotic organisms is very rare. Squalene epoxidase and lanosterol synthase are prerequisite to cyclic sterol biosynthesis. These two enzymes, from the methanotrophic bacterium Methylococcus capsulatus, were functionally expressed in Escherichia coli. Structural analyses of the enzymatic products indicated that the reactions proceeded in a complete regio- and stereospecific fashion to afford (3S)-2,3-oxidosqualene from squalene and lanosterol from (3S)-2,3-oxidosqualene, in full accordance with those of eukaryotes. However, our result obtained with the putative lanosterol synthase was inconsistent with a previous report that the prokaryote accepts both (3R)- and (3S)-2,3-oxidosqualenes to afford 3-epi-lanosterol and lanosterol, respectively. This is the first report demonstrating the existence of the genes encoding squalene epoxidase and lanosterol synthase in prokaryotes by establishing the enzyme activities. The evolutionary aspect of prokaryotic squalene epoxidase and lanosterol synthase is discussed. PMID:17928701

  8. A stopped-flow kinetic study of soluble methane mono-oxygenase from Methylococcus capsulatus (Bath).

    PubMed Central

    Green, J; Dalton, H

    1989-01-01

    1. The roles of the three protein components of soluble methane mono-oxygenase were investigated by the use of rapid-reaction techniques. The transfer of electrons through the enzyme complex from NADH to methane/O2 was also investigated. 2. Electron transfer from protein C, the reductase component, to protein A, the hydroxylase component, was demonstrated. Protein C was shown to undergo a three-electron--one-electron catalytic cycle. The interaction of protein C with NADH was investigated. Reduction of protein C was shown to be rapid, and a charge-transfer interaction between reduced FAD and NAD+ was observed; this intermediate was also found in static titration experiments. Thus the binding of NADH, the reduction of protein C and the intramolecular transfer of electrons through protein C were shown to be much more rapid than the turnover rate of methane mono-oxygenase. 3. The rate of transfer of electrons from protein C to protein A was shown to be lower than the reduction of protein C but higher than the turnover rate of methane mono-oxygenase. Association of the proteins was not rate-limiting. The amount of protein A present in the system had a small effect on the rate of reduction of protein C, indicating some co-operativity between the two proteins. 4. Protein B was shown to prevent electron transfer between protein C and protein A in the absence of methane. On addition of saturating concentrations of methane electron transfer was restored. With saturating concentrations of methane and O2 the observed rate constant for the conversion of methane into methanol was 0.26 s-1 at 18 degrees C. 5. By the use of [2H4]methane it was demonstrated that C-H-bond breakage is likely to be the rate-limiting step in the conversion of methane into methanol. PMID:2497729

  9. Characterization and structural analysis of an active particulate methane monooxygenase trimer from Methylococcus capsulatus (Bath).

    PubMed

    Kitmitto, Ashraf; Myronova, Natalia; Basu, Piku; Dalton, Howard

    2005-08-23

    The oxidation of methane to methanol in methanotrophs is catalyzed by the enzyme methane monooxygenase (MMO). Two distinct forms of this enzyme exist, a soluble cytoplasmic MMO (sMMO) and a membrane-bound particulate form (pMMO). We describe here the biochemical characterization of a stable and active purified pMMO hydroxylase (pMMO-H) and report a three-dimensional (3D) structure, determined by electron microscopy and single-particle analysis at 23 A resolution. Both biochemical and structural data indicate that pMMO hydroxylase is trimeric, with each monomer unit comprised of three polypeptides of 47, 26, and 23 kDa. Comparison of the recent crystal structure [Lieberman, R. L., and Rosenzweig, A. C. (2005) Nature 434, 177] of an uncharacterized pMMO-H complex with the three-dimensional (3D) structure determined here yielded a good match between the principal features and the organization of the enzyme monomers into trimers. The data presented here advance our current understanding of particulate methane monooxygenase function by the characterization of an active form of the enzyme and the corresponding 3D structure. PMID:16101279

  10. Preparation and characterization of a (Cu,Zn)-pMMO from Methylococcus capsulatus (Bath).

    PubMed

    Chen, Chang-Li; Chen, Kelvin H-C; Ke, Shyue-Chu; Yu, Steve S-F; Chan, Sunney I

    2004-12-01

    We report the preparation of a (Cu,Zn)-particulate methane monooxygenase (pMMO) in which the bulk of the copper ions of the electron-transfer clusters (E-clusters) has been replaced by divalent Zn ions. The Cu and Zn contents in the (Cu,Zn)-pMMO were determined by both inductively coupled plasma mass spectroscopy (ICP-MS) and X-ray absorption K-edge spectroscopy. Further characterization of the (Cu,Zn)-pMMO was provided by pMMO-activity assays as well as low-temperature electron paramagnetic resonance (EPR) spectroscopy following reductive titration and incubation in air or air/propylene mixtures. The pMMO-activity assays indicated that the (Cu,Zn)-pMMO was no longer capable of supporting catalytic turnover of hydrocarbon substrates. However, the EPR studies revealed that the catalytic cluster (C-cluster) copper ions in the (Cu,Zn)-pMMO were still capable of supporting the activation of dioxygen when reduced, and that the 14N-superhyperfine features associated with one of the type 2 Cu(II) centers in the hydroxylation C-cluster remained unperturbed. The replacement of the E-cluster copper ions by Zn ions did compromise the ability of the protein to mediate the transfer of reducing equivalents from exogenous reductants to the C-clusters. These observations provide strong support for the electron transfer and catalytic roles for the E-cluster and C-cluster copper ions, respectively. PMID:15541502

  11. Overexpression and purification of the particulate methane monooxygenase from Methylococcus capsulatus (Bath).

    PubMed

    Chan, Sunney I; Nguyen, H-Hoa T; Chen, Kelvin H-C; Yu, Steve S-F

    2011-01-01

    The particulate methane monooxygenase (pMMO) is a multi-copper enzyme that mediates the facile conversion of methane to methanol in methanotrophic bacteria. As a membrane-bound multi-subunit metalloprotein, the highly active protein has been difficult to isolate and purify to homogeneity for biochemical and biophysical studies. In this chapter, we describe a method to overexpress pMMO with good specific activity in high yields in the intracytoplasmic membranes of the host organism, together with two protocols to isolate and purify the enzyme from pMMO-enriched membranes without loss of the copper cofactors and enzymatic activity. PMID:21419922

  12. Concerted mechanism for ethane hydroxylation by the particulate methane monooxygenase from Methylococcus capsulatus (Bath)

    SciTech Connect

    Wilkinson, B.; Priestly, N.D.; Floss, H.G.; Zhu, M.; Nguyen, H.H.T.; Chan, S.I.; Morimoto, Hiromi; Williams, P.G.

    1996-01-31

    The ethanol samples in the isolated alcohol/water mixtures were converted into their (2R)-2-acetoxy-2-phenylethanoate derivatives (2-34 mCi). Examination of the well-resolved {sup 3}H NMR spectra for these derivatives produced an exceptionally consistent set of stereochemical data. When corrected for the enantiomeric purity of the ethyl tosylate starting materials, the data clearly show that the reaction occurs with complete retention of configuration, i.e., with 100% stereoselection. Barring substantial slowing of the carbon-carbon bond rotation of the ethyl radical when bound to the enzyme, this result rules out mechanisms proceeding via alkyl radical (and/or cation) structures, even very short-lived ones, as such intermediates would have to have a lifetime of < 1 x 10{sup -14} s in order not to undergo any detectable C-C bond rotation, i.e., the capture reaction would have to be much faster than the decay of a transition state. The data instead point to a mechanism in which C-H bond cleavage is preceded by bond formation at the alkyl carbon, i.e., one proceeding through a pentacoordinated carbon species. 29 refs., 1 fig., 1 tab.

  13. Oxidation of C1 Compounds by Particulate fractions from Methylococcus capsulatus: distribution and properties of methane-dependent reduced nicotinamide adenine dinucleotide oxidase (methane hydroxylase).

    PubMed Central

    Ribbons, D W

    1975-01-01

    Cell-free particulate fractions of extracts from the obligate methylotroph Methylococcus capsulatus catalyze the reduced nicotinamide adenine dinucleotide (NADH) and O2-dependent oxidation of methane (methane hydroxylase). The only oxidation product detected was formate. These preparations also catalyze the oxidation of methanol and formaldehyde to formate in the presence or absence of phenazine methosulphate with oxygen as the terminal electron acceptor. Methane hydroxylase activity cannot be reproducibly obtained from disintegrated cell suspensions even though the whole cells actively respired when methane was presented as a substrate. Varying the disintegration method or extraction medium had no significant effect on the activities obtained. When active particles were obtained, hydroxylase activity was stable at 0 C for days. Methane hydroxylase assays were made by measuring the methane-dependent oxidation of NADH by O2. In separate experiments, methane consumption and the accumulation of formate were also demonstrated. Formate is not oxidized by these particulate fractions. The effects of particle concentration, temperature, pH, and phosphate concentration on enzymic activity are described. Ethane is utilized in the presence of NADH and O2. The stoichiometric relationships of the reaction(s) with methane as substrate were not established since (i) the presumed initial product, methanol, is also oxidized to formate, and (ii) the contribution that NADH oxidase activity makes to the observed consumption of reactants could not be assessed in the presence of methane. Studies with known inhibitors of electron transport systems indicate that the path of electron flow from NADH to oxygen is different for the NADH oxidase, methane hydroxylase, and methanol oxidase activities. Images PMID:238946

  14. Copper-containing protein from the membranes of methane oxidizing bacterium Methylococcus capsulatus (strain M) containing methane monooxygenase

    SciTech Connect

    Burbaev, D.Sh.; Moroz, I.A.; Gvozdev, R.I. |

    1994-12-31

    The goal of the present work was to study copper-containing center of the membrane of methane oxidizing bacterium, M. capsulatus, strain M, by ESR spectroscopy. The bacteria were grown and the membrane preparation particles, fraction O{sub 1} were isolated as described earlier. The results reveal that the fraction of particles mediating oxidation of CH{sub 4} includes a Cu-protein with a minimal molecular mass of 49 kD. This protein has the type 2 ESR signal characteristic of copper with nitrogen-containing ligands. Histidine residues are most probable ligands. The protein is likely to be incorporated into pMMO, although its function (electron transfer, activation of {sub 2}) is far for clear.

  15. Biochemical characterization of the water-soluble squalene synthase from Methylococcus capsulatus and the functional analyses of its two DXXD(E)D motifs and the highly conserved aromatic amino acid residues.

    PubMed

    Ohtake, Kana; Saito, Naoki; Shibuya, Satoshi; Kobayashi, Wakako; Amano, Ryosuke; Hirai, Takumi; Sasaki, Shinji; Nakano, Chiaki; Hoshino, Tsutomu

    2014-12-01

    Information regarding squalene synthases (SQSs) from prokaryotes is scarce. We aimed to characterize the SQS from Methylococcus capsulatus. We studied its reaction mechanism by kinetic analysis and evaluated the structure of the substrate/inhibitor-binding sites via homology modeling. The cloned M. capsulatus SQS was expressed in Escherichia coli and purified by nickel-nitrilotriacetic acid column chromatography. Interestingly, M. capsulatus SQS was water-soluble and did not require any detergent for its higher activity, unlike other SQSs studied previously; supplementation of any type of detergent inhibited enzyme activity. The specific activity and the kinetic values (Km and kcat ) for the substrate farnesyl diphosphate and NADPH are reported. The substrate analog farnesyl methylenediphosphonate showed potent inhibition toward the enzyme. We prepared the site-specific mutants directed at potential active-site residues (58) DXX(61) E(62) D (S1 site) and (213) DXX(216) D(217) D (S2 site), which were assumed to be involved in the binding of the substrate farnesyl diphosphate through the Mg(2+) ion. We first demonstrated that the S1 site and the two basic residues (R55 and K212) were responsible for the binding of farnesyl diphosphate. Furthermore, we examined the catalytic roles of the highly conserved aromatic residues and demonstrated that the Y164 residue abstracts the proton of cation 5, which is produced during the first half-reaction (Scheme 1), to afford presqualene diphosphate, and that the W224 residue stabilizes the intermediary cation 5 via the cation-π interaction. Furthermore, we confirm for the first time that the F32 and the Y51 residues also stabilize the carbocation intermediate(s) generated during the second half-reaction. PMID:25283713

  16. Product bound structures of the soluble methane monooxygenase hydroxylase from Methylococcus capsulatus (Bath): protein motion in the alpha-subunit.

    PubMed

    Sazinsky, Matthew H; Lippard, Stephen J

    2005-04-27

    The soluble methane monooxygenase hydroxylase (MMOH) alpha-subunit contains a series of cavities that delineate the route of substrate entrance to and product egress from the buried carboxylate-bridged diiron center. The presence of discrete cavities is a major structural difference between MMOH, which can hydroxylate methane, and toluene/o-xylene monooxygenase hydroxylase (ToMOH), which cannot. To understand better the functions of the cavities and to investigate how an enzyme designed for methane hydroxylation can also accommodate larger substrates such as octane, methylcubane, and trans-1-methyl-2-phenylcyclopropane, MMOH crystals were soaked with an assortment of different alcohols and their X-ray structures were solved to 1.8-2.4 A resolution. The product analogues localize to cavities 1-3 and delineate a path of product exit and/or substrate entrance from the active site to the surface of the protein. The binding of the alcohols to a position bridging the two iron atoms in cavity 1 extends and validates previous crystallographic, spectroscopic, and computational work indicating this site to be where substrates are hydroxylated and products form. The presence of these alcohols induces perturbations in the amino acid side-chain gates linking pairs of cavities, allowing for the formation of a channel similar to one observed in ToMOH. Upon binding of 6-bromohexan-1-ol, the pi helix formed by residues 202-211 in helix E of the alpha-subunit is extended through residue 216, changing the orientations of several amino acid residues in the active site cavity. This remarkable secondary structure rearrangement in the four-helix bundle has several mechanistic implications for substrate accommodation and the function of the effector protein, MMOB. PMID:15839679

  17. Mössbauer studies of the membrane-associated methane monooxygenase from Methylococcus capsulatus bath: evidence for a Diiron center.

    PubMed

    Martinho, Marlène; Choi, Dong W; Dispirito, Alan A; Antholine, William E; Semrau, Jeremy D; Münck, Eckard

    2007-12-26

    Two methane monooxygenase (MMO) systems have been identified in methanotrophic bacteria, namely, a soluble or cytoplasmic MMO and a membrane-associated or particulate MMO. The active site of the well-characterized soluble MMO contains a bis-mu-hydroxo-bridged diiron cluster. X-ray crystallographic studies of the particulate enzyme, pMMO, have identified two copper centers on the alpha subunit (pmoB) of the alphabetagamma trimer and a site at the interface of the betagamma subunits filled by a Zn, apparently from the crystallization buffer. In our hands, pMMO preparations containing 1-2 iron atoms per alphabetagamma show the highest catalytic activity. We have employed Mössbauer spectroscopy to characterize the iron in our preparations. Interestingly, we find in pMMO a component with the same spectral properties as the antiferromagnetically coupled diiron(III) cluster in the soluble enzyme. In whole cells, we find nearly 1 diiron center per alphabetagamma of pMMO; in purified enzyme preparations, only 10% of the sites appear to be occupied. These occupancies correlate well with the measured specific activities of purified pMMO and pMMO in whole cells. We suggest that it is the "Zn site" that accommodates the diiron center in active pMMO. PMID:18052283

  18. Spectroscopic detection of intermediates in the reaction of dioxygen with the reduced methane monooxygenase hydroxylase from Methylococcus capsulatus (bath)

    SciTech Connect

    Liu, K.E.; Salifoglou, A.; Lippard, S.J. ); Wang, D.; Huynh, B.H.; Edmondson, D.E. )

    1994-08-10

    Soluble methane monoxygenase (MMO), an enzyme system isolated from methanotrophic bacteria, which catalyzes the conversion of methane to methanol, has three components, a hydroxylase (H), a coupling protein (B), and a reductase (R). In this communication we present the results of stopped-flow and freeze-quench kinetic investigations of the reaction of dioxygen with the reduced hydroxylase, H[sub red], which elaborate upon and extend the findings of parallel work on MMOH from Methylosinus trichosporium OB3b. 34 refs., 1 fig.

  19. Characterization of the second prosthetic group of the flavoenzyme NADH-acceptor reductase (component C) of the methane mono-oxygenase from Methylococcus capsulatus (Bath).

    PubMed Central

    Colby, J; Dalton, H

    1979-01-01

    1. A new two-step purification is described that routinely yields 100mg quantities of component C for biochemical studies. 2. Chemical analyses show component C purified by this procedure to contain 2 g-atoms of iron, 2 mol of acid-labile sulphide (S) and 1 mol of FAD per mol of protein. 3. The Fe-S core of component C was extruded by treating the protein with p-methoxybenzenethiol in hexamethyl phosphoramide/50mM-Tris/HCl buffer, pH 8.5 (4:1, v/v), under anaerobic conditions. The spectral properties of the extruded core suggest that component C contains 1 mol of [2Fe-2S(S-Cys)4] centre per mol of protein. 4. E.p.r. spectroscopy confirms the presence of a Fe-S centre in component C. 5. Component C catalyses the reduction by NADH of ferricyanide, 2,6-dichlorophenol-indophenol or horse heart cytochrome c, with specific activities of 50--230 units/mg of protein. 6. The optimum pH for the NADH-acceptor reductase activity is 8.5--9.0, and the apparent Km values for NADH and NADPH are 0.05mM and 15.5mM respectively. 7. Unlike methane mono-oxygenase activity, NADH-acceptor reductase activity of component C is not inhibited by 8-hydroxyquinoline or by acetylene. PMID:220953

  20. X-ray absorption and EPR studies on the copper ions associated with the particulate methane monooxygenase from Methylococcus capsulatus (Bath). Cu(I) ions and their implications

    SciTech Connect

    Nguyen, H.H.T.; Elliott, S.J.; Lidstrom, M.E.; Chan, S.I.; Nakagawa, K.H.; Hedman, B.; Hodgson, K.O.

    1996-12-18

    Parallel X-ray absorption edge and EPR studies of the particulate methane monooxygenase in situ reveal that the enzyme contains unusually high levels of copper ions with a significant portion of the copper ions existing as Cu(I) in the `as-isolated` form (70-80%). The observation of high levels of reduced copper in a monooxygenase is surprising considering that the natural cosubstrate of the enzyme is dioxygen. Toward clarifying the roles of the various copper ions in the enzyme, we have successfully prepared different states of the protein in the membrane-bound form at various levels of reduction using dithionite, dioxygen, and ferricyanide. EPR intensity analysis of the fully-oxidized preparations indicates that the bulk of copper ions are arranged in cluster units. The fully-reduced protein obtained by reduction by dithionite has been used to initiate the single turnover of the enzyme in the presence of dioxygen. Differential reactivity toward dioxygen was observed upon analyzing the copper reduction levels in these synchronized preparations. The enzyme is capable of supporting turnover in the absence of external electron donors in the highly reduced states. These results suggest the presence of at least two classes of copper ions in the particulate methane monoxygenase. As a working hypothesis, we have referred to these classes of copper ions as (1) the catalytic (C) clusters, and (2) the electron transfer (E) clusters. 56 refs., 4 figs., 1 tab.

  1. Revisiting the mechanism of dioxygen activation in soluble methane monooxygenase from M. capsulatus (Bath): evidence for a multi-step, proton-dependent reaction pathway.

    PubMed

    Tinberg, Christine E; Lippard, Stephen J

    2009-12-29

    Stopped-flow kinetic investigations of soluble methane monooxygenase (sMMO) from M. capsulatus (Bath) have clarified discrepancies that exist in the literature regarding several aspects of catalysis by this enzyme. The development of thorough kinetic analytical techniques has led to the discovery of two novel oxygenated iron species that accumulate in addition to the well-established intermediates H(peroxo) and Q. The first intermediate, P*, is a precursor to H(peroxo) and was identified when the reaction of reduced MMOH and MMOB with O(2) was carried out in the presence of >or=540 microM methane to suppress the dominating absorbance signal due to Q. The optical properties of P* are similar to those of H(peroxo), with epsilon(420) = 3500 M(-1) cm(-1) and epsilon(720) = 1250 M(-1) cm(-1). These values are suggestive of a peroxo-to-iron(III) charge-transfer transition and resemble those of peroxodiiron(III) intermediates characterized in other carboxylate-bridged diiron proteins and synthetic model complexes. The second identified intermediate, Q*, forms on the pathway of Q decay when reactions are performed in the absence of hydrocarbon substrate. Q* does not react with methane, forms independently of buffer composition, and displays a unique shoulder at 455 nm in its optical spectrum. Studies conducted at different pH values reveal that rate constants corresponding to P* decay/H(peroxo) formation and H(peroxo) decay/Q formation are both significantly retarded at high pH and indicate that both events require proton transfer. The processes exhibit normal kinetic solvent isotope effects (KSIEs) of 2.0 and 1.8, respectively, when the reactions are performed in D(2)O. Mechanisms are proposed to account for the observations of these novel intermediates and the proton dependencies of P* to H(peroxo) and H(peroxo) to Q conversion. PMID:19921958

  2. Bathing

    MedlinePlus

    ... in the tub or shower. • Always check the water temperature before he or she gets in the tub ... to do, step by step. • Make sure the water temperature is comfortable. • Don’t use bath oil. It ...

  3. A continuous-wave electron-nuclear double resonance (X-band) study of the Cu2+ sites of particulate methane mono-oxygenase of Methylococcus capsulatus (strain M) in membrane and pure dopamine beta-mono-oxygenase of the adrenal medulla.

    PubMed

    Katterle, Bettina; Gvozdev, Rudolf I; Abudu, Ntei; Ljones, Torbjørn; Andersson, K Kristoffer

    2002-05-01

    All methanotrophic bacteria express a membrane-bound (particulate) methane mono-oxygenase (pMMO). In the present study, we have investigated pMMO in membrane fragments from Methylococcus capsulatus (strain M). pMMO contains a typical type-2 Cu(2+) centre with the following EPR parameters: g(z) 2.24, g(x,y) 2.06, A(Cu)(z) 19.0 mT and A(Cu)(x,y) 1.0 mT. Simulation of the Cu(2+) spectrum yielded a best match by using four equivalent nitrogens (A(N)=1.5 mT, 42 MHz). Incubation with ferricyanide neither changed nor increased the amount of EPR-active Cu(2+), in contrast with other reports. The EPR visible copper seems not to be part of any cluster, as judged from the microwave power saturation behaviour. Continuous-wave electron-nuclear double resonance (CW ENDOR; 9.4 GHz, 5-20 K) experiments at g( perpendicular) of the Cu(II) spectrum show a weak coupling to protons with an A(H) of 2.9 MHz that corresponds to a distance of 3.8 A (1 A identical with 0.1 nm), assuming that it is a purely dipolar coupling. Incubation in (2)H(2)O leads to a significant decrease in these (1)H-ENDOR intensities, showing that these protons are exchangeable. This result strongly suggests that the EPR visible copper site of pMMO is accessible to solvent, which was confirmed by the chelation of the Cu(2+) by diethyldithiocarbamic acid. The (1)H and (14)N hyperfine coupling constants confirm a histidine ligation of the EPR visible copper site in pMMO. The hyperfine structure in the ENDOR or EPR spectra of pMMO is not influenced by the inhibitors azide, cyanide or ammonia, indicating that they do not bind to the EPR visible copper. We compared pMMO with the type-2 Cu(2+) enzyme, dopamine beta-mono-oxygenase (DbetaM). For DbetaM, it is assumed that the copper site is solvent-accessible. CW ENDOR shows similar weakly coupled and (2)H(2)O-exchangeable protons (2.9 MHz), as observed in pMMO, as well as the strongly coupled nitrogens (40 MHz) from the co-ordinating N of the histidines in DbetaM. In

  4. A continuous-wave electron-nuclear double resonance (X-band) study of the Cu2+ sites of particulate methane mono-oxygenase of Methylococcus capsulatus (strain M) in membrane and pure dopamine beta-mono-oxygenase of the adrenal medulla.

    PubMed Central

    Katterle, Bettina; Gvozdev, Rudolf I; Abudu, Ntei; Ljones, Torbjørn; Andersson, K Kristoffer

    2002-01-01

    All methanotrophic bacteria express a membrane-bound (particulate) methane mono-oxygenase (pMMO). In the present study, we have investigated pMMO in membrane fragments from Methylococcus capsulatus (strain M). pMMO contains a typical type-2 Cu(2+) centre with the following EPR parameters: g(z) 2.24, g(x,y) 2.06, A(Cu)(z) 19.0 mT and A(Cu)(x,y) 1.0 mT. Simulation of the Cu(2+) spectrum yielded a best match by using four equivalent nitrogens (A(N)=1.5 mT, 42 MHz). Incubation with ferricyanide neither changed nor increased the amount of EPR-active Cu(2+), in contrast with other reports. The EPR visible copper seems not to be part of any cluster, as judged from the microwave power saturation behaviour. Continuous-wave electron-nuclear double resonance (CW ENDOR; 9.4 GHz, 5-20 K) experiments at g( perpendicular) of the Cu(II) spectrum show a weak coupling to protons with an A(H) of 2.9 MHz that corresponds to a distance of 3.8 A (1 A identical with 0.1 nm), assuming that it is a purely dipolar coupling. Incubation in (2)H(2)O leads to a significant decrease in these (1)H-ENDOR intensities, showing that these protons are exchangeable. This result strongly suggests that the EPR visible copper site of pMMO is accessible to solvent, which was confirmed by the chelation of the Cu(2+) by diethyldithiocarbamic acid. The (1)H and (14)N hyperfine coupling constants confirm a histidine ligation of the EPR visible copper site in pMMO. The hyperfine structure in the ENDOR or EPR spectra of pMMO is not influenced by the inhibitors azide, cyanide or ammonia, indicating that they do not bind to the EPR visible copper. We compared pMMO with the type-2 Cu(2+) enzyme, dopamine beta-mono-oxygenase (DbetaM). For DbetaM, it is assumed that the copper site is solvent-accessible. CW ENDOR shows similar weakly coupled and (2)H(2)O-exchangeable protons (2.9 MHz), as observed in pMMO, as well as the strongly coupled nitrogens (40 MHz) from the co-ordinating N of the histidines in DbetaM. In

  5. Inhibition of methane oxidation by Methylococcus capsulatus with hydrochlorofluorocarbons and fluorinated methanes

    SciTech Connect

    Matheson, L.J.; Oremland, R.S.; Jahnke, L.L.

    1997-07-01

    Concerns about stratospheric ozone and global warming have focused some inquiries upon the microbial degradation of some atmospheric halocarbons. Little is known about the interaction of hydrochlorofluorocarbons (HCFCs) and hydrofluorocarbons (HFCs). This study examines possible interactions, including the inhibition of methane oxidation by chlorinated solvents, whether oxidation products formed may have inhibitory effects of their own, and whether other fluorinated methanes inhibit methane oxidation by whole cells. 33 refs., 4 figs., 1 tab.

  6. Bubble bath soap poisoning

    MedlinePlus

    ... medlineplus.gov/ency/article/002762.htm Bubble bath soap poisoning To use the sharing features on this page, please enable JavaScript. Bubble bath soap poisoning occurs when someone swallows bubble bath soap. ...

  7. Polyhydroxyalkanoate production in Rhodobacter capsulatus: genes, mutants, expression, and physiology.

    PubMed Central

    Kranz, R G; Gabbert, K K; Locke, T A; Madigan, M T

    1997-01-01

    Like many other prokaryotes, the photosynthetic bacterium Rhodobacter capsulatus produces high levels of polyhydroxyalkanoates (PHAs) when a suitable carbon source is available. The three genes that are traditionally considered to be necessary in the PHA biosynthetic pathway, phaA (beta-ketothiolase), phaB (acetoacetylcoenzyme A reductase), and phaC (PHA synthase), were cloned from Rhodobacter capsulatus. In R. capsulatus, the phaAB genes are not linked to the phaC gene. Translational beta-galactosidase fusions to phaA and phaC were constructed and recombined into the chromosome. Both phaC and phaA were constitutively expressed regardless of whether PHA production was induced, suggesting that control is posttranslational at the enzymatic level. Consistent with this conclusion, it was shown that the R. capsulatus transcriptional nitrogen-sensing circuits were not involved in PHA synthesis. The doubling times of R. capsulatus transcriptional nitrogen-sensing circuits were not involved in PHA synthesis. The doubling times of R. capsulatus grown on numerous carbon sources were determined, indicating that this bacterium grows on C2 to C12 fatty acids. Grown on acetone, caproate, or heptanoate, wild-type R. capsulatus produced high levels of PHAs. Although a phaC deletion strain was unable to synthesize PHAs on any carbon source, phaA and phaAB deletion strains were able to produce PHAs, indicating that alternative routes for the synthesis of substrates for the synthase are present. The nutritional versatility and bioenergetic versatility of R. capsulatus, coupled with its ability to produce large amounts of PHAs and its genetic tractability, make it an attractive model for the study of PHA production. PMID:9251189

  8. What Are Bath Salts?

    MedlinePlus

    ... Are bath salts becoming more popular? Marsha Lopez Hi, Lauren. Nope! Actually quite the opposite! This family ... and how dangerous for your body? Michelle Rankin Hi ParkerPanella - Bath salts are drugs known as synthetic ...

  9. METAL COATING BATHS

    DOEpatents

    Robinson, J.W.

    1958-08-26

    A method is presented for restoring the effectiveness of bronze coating baths used for hot dip coating of uranium. Such baths, containing a high proportion of copper, lose their ability to wet uranium surfaces after a period of use. The ability of such a bath to wet uranium can be restored by adding a small amount of metallic aluminum to the bath, and skimming the resultant hard alloy from the surface.

  10. Secretion of flavins by three species of methanotrophic bacteria.

    PubMed

    Balasubramanian, Ramakrishnan; Levinson, Benjamin T; Rosenzweig, Amy C

    2010-11-01

    We detected flavins in the growth medium of the methanotrophic bacterium Methylocystis species strain M. Flavin secretion correlates with growth stage and increases under iron starvation conditions. Two other methanotrophs, Methylosinus trichosporium OB3b and Methylococcus capsulatus (Bath), secrete flavins, suggesting that flavin secretion may be common to many methanotrophic bacteria. PMID:20833792

  11. Complete Genome Sequences of Five Bacteriophages That Infect Rhodobacter capsulatus.

    PubMed

    Bollivar, David W; Bernardoni, Brooke; Bockman, Matthew R; Miller, Brenda M; Russell, Daniel A; Delesalle, Veronique A; Krukonis, Gregory P; Hatfull, Graham F; Cross, Madeline R; Szewczyk, Marlena M; Eppurath, Atul

    2016-01-01

    Five bacteriophages that infect the Rhodobacter capsulatus strain YW1 were isolated from stream water near Bloomington, Illinois, USA. Two distinct genome types are represented in the newly isolated bacteriophages. These genomes are different from other bacteriophage genomes previously described. PMID:27231352

  12. Complete Genome Sequences of Five Bacteriophages That Infect Rhodobacter capsulatus

    PubMed Central

    Bernardoni, Brooke; Bockman, Matthew R.; Miller, Brenda M.; Russell, Daniel A.; Delesalle, Veronique A.; Krukonis, Gregory P.; Hatfull, Graham F.; Cross, Madeline R.; Szewczyk, Marlena M.; Eppurath, Atul

    2016-01-01

    Five bacteriophages that infect the Rhodobacter capsulatus strain YW1 were isolated from stream water near Bloomington, Illinois, USA. Two distinct genome types are represented in the newly isolated bacteriophages. These genomes are different from other bacteriophage genomes previously described. PMID:27231352

  13. Physical map of the genome of Rhodobacter capsulatus SB 1003.

    PubMed Central

    Fonstein, M; Zheng, S; Haselkorn, R

    1992-01-01

    A map of the chromosome of Rhodobacter capsulatus was constructed by overlapping the large restriction fragments generated by endonucleases AseI and XbaI. The analyses were done by hybridization of single fragments with the restriction fragments blotted from pulsed-field gels and by grouping cosmids of a genomic library of R. capsulatus into contigs, corresponding to the restriction fragments, and further overlapping of the contigs. A technical difficulty due to a repeated sequence made it necessary to use hybridization with cloned genes and prior knowledge of the genetic map in order to close the physical circle in a unique way. In all, 41 restriction sites were mapped on the 3.6-Mb circular genome and 22 genes were positioned at 26 loci of the map. Cosmid clones were grouped in about 80 subcontigs, forming two groups, one corresponding to the chromosome of R. capsulatus and the other corresponding to a 134-kb plasmid. cos site end labeling and partial digestion of cosmids were used to construct a high-resolution EcoRV map of the 134-kb plasmid. The same method can be extended to the entire chromosome. The cosmid clones derived in this work can be used as a hybridization panel for the physical mapping of new genes as soon as they are cloned. Images PMID:1597421

  14. Effect of dietary Rhodobacter capsulatus on egg-yolk cholesterol and laying hen performance.

    PubMed

    Salma, U; Miah, A G; Tareq, K M A; Maki, T; Tsujii, H

    2007-04-01

    The present study was conducted to investigate the effects of dietary Rhodobacter capsulatus on the laying hen. A total of forty 23-wk-old Hy-Line Brown laying hens were randomly assigned into 4 treatment groups (10 laying hens/group) and fed diets supplemented with 0 (control), 0.01, 0.02, and 0.04% R. capsulatus during the 60-d feeding period. Dietary supplementation of R. capsulatus (0.04%) reduced (P < 0.05) cholesterol and triglycerides concentration in serum (15 and 11%), as well as in egg-yolk (13 and 16%) over a 60-d feeding period. Cholesterol and triglycerides concentrations in serum as well as egg-yolk were changed linearly in accordance with increasing levels of dietary R. capsulatus. Supplementation of R. capsulatus in diets increased high-density lipoprotein cholesterol level and decreased (P < 0.05) atherogenic index in serum. Yolk color was improved (P < 0.05) in the group fed the 0.04% R. capsulatus supplemented diet compared with the control group. Hepatic cholesterol and triglycerides were reduced (P < 0.05) by 0.04% R. capsulatus. Moreover, the supplementation of R. capsulatus in layer diets did not appear to cause any adverse effects on egg production, shell weight, shell thickness, Haugh unit, yolk index, and feed conversion efficiency compared with the same parameters for the control laying hens. It is postulated that known and unknown factors are present in R. capsulatus presumably responsible for the hypocholesterolemic effect on laying hens. Therefore, the dietary supplementation of R. capsulatus may lead to the development of low-cholesterol chicken eggs as demanded by health-conscious consumers. PMID:17369543

  15. Characterization of a diiron(III) peroxo intermediate in the reaction cycle of methane monooxygenase hydroxylase from Methylococcus cupsulatus (Bath)

    SciTech Connect

    Liu, K.E.; Valentine, A.M.; Lippard, S.J.; Qiu, D.; Spiro, T.G.; Edmondson, D.E.; Appelman, E.H.

    1995-05-03

    We present new optical and resonance Raman spectroscopic data that characterize the first intermediate as a diiron(III) peroxo species. The time-resolved optical and freeze-quench resonance Raman spectroscopic experiments strongly support assignment of the first intermediate, variously designated P or L, in the MMO hydroxylase reaction cycle as a symmetrical diiron(III) peroxo species. 32 refs., 2 figs.

  16. [Immersion in a bath despite a safety bath chair].

    PubMed

    Christensen, H B; Lange, A

    1989-01-01

    A case of submersion is described. A mother left her child aged 8 1/2 months sitting in a "safety bath chair" in a full bath and found the child lying under the water shortly afterwards. The infant was hypotonic for a brief period but rapidly recovered without sequelae. Use of a "safety bath chair" gives a false sense of security and its use is warned against. PMID:2911907

  17. 33 CFR 165.104 - Safety Zone: Vessel Launches, Bath Iron Works, Kennebec River, Bath, Maine.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ..., Bath Iron Works, Kennebec River, Bath, Maine. 165.104 Section 165.104 Navigation and Navigable Waters... Guard District § 165.104 Safety Zone: Vessel Launches, Bath Iron Works, Kennebec River, Bath, Maine. (a... Bath Iron Works dry dock while it is being moved to and from its moored position at the Bath Iron...

  18. 33 CFR 165.104 - Safety Zone: Vessel Launches, Bath Iron Works, Kennebec River, Bath, Maine.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ..., Bath Iron Works, Kennebec River, Bath, Maine. 165.104 Section 165.104 Navigation and Navigable Waters... Guard District § 165.104 Safety Zone: Vessel Launches, Bath Iron Works, Kennebec River, Bath, Maine. (a... Bath Iron Works dry dock while it is being moved to and from its moored position at the Bath Iron...

  19. 33 CFR 165.104 - Safety Zone: Vessel Launches, Bath Iron Works, Kennebec River, Bath, Maine.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ..., Bath Iron Works, Kennebec River, Bath, Maine. 165.104 Section 165.104 Navigation and Navigable Waters... Guard District § 165.104 Safety Zone: Vessel Launches, Bath Iron Works, Kennebec River, Bath, Maine. (a... Bath Iron Works dry dock while it is being moved to and from its moored position at the Bath Iron...

  20. 33 CFR 165.104 - Safety Zone: Vessel Launches, Bath Iron Works, Kennebec River, Bath, Maine.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ..., Bath Iron Works, Kennebec River, Bath, Maine. 165.104 Section 165.104 Navigation and Navigable Waters... Guard District § 165.104 Safety Zone: Vessel Launches, Bath Iron Works, Kennebec River, Bath, Maine. (a... Bath Iron Works dry dock while it is being moved to and from its moored position at the Bath Iron...

  1. 33 CFR 165.104 - Safety Zone: Vessel Launches, Bath Iron Works, Kennebec River, Bath, Maine.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ..., Bath Iron Works, Kennebec River, Bath, Maine. 165.104 Section 165.104 Navigation and Navigable Waters... Guard District § 165.104 Safety Zone: Vessel Launches, Bath Iron Works, Kennebec River, Bath, Maine. (a... Bath Iron Works dry dock while it is being moved to and from its moored position at the Bath Iron...

  2. Cytochromes P460 and c'-beta; a new family of high-spin cytochromes c.

    PubMed

    Elmore, Bradley O; Bergmann, David J; Klotz, Martin G; Hooper, Alan B

    2007-03-01

    Cytochromes-P460 of Nitrosomonas europaea and Methylococcus capsulatus (Bath), and the cytochrome c' of M. capsulatus, believed to be involved in binding or transformation of N-oxides, are shown to represent an evolutionarily related new family of monoheme, approximately 17kDa, cytochromes c found in the genomes of diverse Proteobacteria. All members of this family have a predicted secondary structure predominantly of beta-sheets in contrast to the predominantly alpha-helical cytochromes c' found in photoheterotrophic and denitrifying Proteobacteria. PMID:17292891

  3. Chlorhexidine gluconate: to bathe or not to bathe?

    PubMed

    Rubin, Caroline; Louthan, Rufina Bavin; Wessels, Erica; McGowan, Mary-Bridgid; Downer, Shantee; Maiden, Jeanne

    2013-01-01

    Despite infection-prevention initiatives, hospital-acquired infections (HAIs) are still a common occurrence. Chlorhexidine gluconate (CHG) is an important antibacterial agent. Research indicates that the intervention of bathing with CHG can reduce the number of HAIs. Chlorhexidine gluconate is known to reduce the bioload of several bacteria, including multiple strains of methicillin-resistant Staphylococcus aureus. Research regarding the intervention of bathing with CHG was assessed and found to reduce central line-related blood stream infections, ventilator-associated pneumonia, and vancomycin-resistant enterococci. The reduction in HAIs was found to be greater as compared to bathing with soap and water. The reduction of these HAIs will allow for a saving of resources, finances and staff time, which may ultimately be passed on to the patient. While further research is indicated, a strong conclusion is drawn that bathing with CHG reduces the number of HAIs. PMID:23470709

  4. Grooming, Bathing and Safety Tips

    MedlinePlus

    ... Wet One of the most common problems that amputees encounter is maintaining balance while bathing and climbing ... 18/2014 Back to Top © Copyrighted by the Amputee Coalition . Local reproduction for use by Amputee Coalition ...

  5. The Rhodobacter capsulatus glnB gene is regulated by NtrC at tandem rpoN-independent promoters.

    PubMed Central

    Foster-Hartnett, D; Kranz, R G

    1994-01-01

    The protein encoded by glnB of Rhodobacter capsulatus is part of a nitrogen-sensing cascade which regulates the expression of nitrogen fixation genes (nif). The expression of glnB was studied by using lacZ fusions, primer extension analysis, and in vitro DNase I footprinting. Our results suggest that glnB is transcribed from two promoters, one of which requires the R. capsulatus ntrC gene but is rpoN independent. Another promoter upstream of glnB is repressed by NtrC; purified R. capsulatus NtrC binds to sites that overlap this distal promoter region. Images PMID:8051036

  6. Analysis of the fnrL gene and its function in Rhodobacter capsulatus.

    PubMed Central

    Zeilstra-Ryalls, J H; Gabbert, K; Mouncey, N J; Kaplan, S; Kranz, R G

    1997-01-01

    The fnr gene encodes a regulatory protein involved in the response to oxygen in a variety of bacterial genera. For example, it was previously shown that the anoxygenic, photosynthetic bacterium Rhodobacter sphaeroides requires the fnrL gene for growth under anaerobic, photosynthetic conditions. Additionally, the FnrL protein in R. sphaeroides is required for anaerobic growth in the dark with an alternative electron acceptor, but it is not essential for aerobic growth. In this study, the fnrL locus from Rhodobacter capsulatus was cloned and sequenced. Surprisingly, an R. capsulatus strain with the fnrL gene deleted grows like the wild type under either photosynthetic or aerobic conditions but does not grow anaerobically with alternative electron acceptors such as dimethyl sulfoxide (DMSO) or trimethylamine oxide. It is demonstrated that the c-type cytochrome induced upon anaerobic growth on DMSO is not synthesized in the R. capsulatus fnrL mutant. In contrast to wild-type strains, R. sphaeroides and R. capsulatus fnrL mutants do not synthesize the anaerobically, DMSO-induced reductase. Mechanisms that explain the basis for FnrL function in both organisms are discussed. PMID:9393689

  7. Transcriptional regulation of the Rhodobacter capsulatus response regulator CtrA

    PubMed Central

    Leung, Molly M.; Brimacombe, Cedric A.; Beatty, J. Thomas

    2013-01-01

    The Rhodobacter capsulatus response regulator CtrA controls the expression of 227 genes, some of which are upregulated by both the phosphorylated and unphosphorylated forms of CtrA. Therefore, CtrA concentration alone, regardless of phosphorylation state, may determine expression of downstream genes, yet little is known about the regulation of ctrA in R. capsulatus. In this study we used a ctrA : : lacZ fusion plasmid to study the effects of medium composition, growth conditions and growth phase on R. capsulatus ctrA gene expression. These experiments indicate that ctrA expression is higher when cultures are grown in phototrophic (anaerobic) conditions compared with chemotrophic (aerobic) conditions, and is higher when grown in a minimal medium compared with a rich medium. We used several mutants to investigate possible regulatory pathways, and found that in R. capsulatus ctrA is not autoregulated but is regulated by a quorum-sensing system. The expression of ctrA increased as cell cultures moved through exponential phase and into stationary phase, with high levels of expression persisting long after culture turbidity plateaued. Although this growth phase-dependent pattern of expression was also observed in a quorum-sensing mutant, the magnitude of ctrA expression was about 50 % of the wild-type strain at all phases. Furthermore, reduction of phosphate concentration in the growth medium decreased ctrA expression in a culture density-independent manner, whereas reduction of malic acid (carbon source) or ammonium (nitrogen source) concentration had no effect. The regulation of ctrA expression in R. capsulatus appears to require the coordination of multiple pathways involved in detecting a variety of environmental conditions. PMID:23154973

  8. 33 CFR 334.45 - Kennebec River, Bath Iron Works Shipyard, naval restricted area, Bath, Maine.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 33 Navigation and Navigable Waters 3 2012-07-01 2012-07-01 false Kennebec River, Bath Iron Works Shipyard, naval restricted area, Bath, Maine. 334.45 Section 334.45 Navigation and Navigable Waters CORPS... REGULATIONS § 334.45 Kennebec River, Bath Iron Works Shipyard, naval restricted area, Bath, Maine. (a)...

  9. 33 CFR 334.45 - Kennebec River, Bath Iron Works Shipyard, naval restricted area, Bath, Maine.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 33 Navigation and Navigable Waters 3 2013-07-01 2013-07-01 false Kennebec River, Bath Iron Works Shipyard, naval restricted area, Bath, Maine. 334.45 Section 334.45 Navigation and Navigable Waters CORPS... REGULATIONS § 334.45 Kennebec River, Bath Iron Works Shipyard, naval restricted area, Bath, Maine. (a)...

  10. 33 CFR 334.45 - Kennebec River, Bath Iron Works Shipyard, naval restricted area, Bath, Maine.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 33 Navigation and Navigable Waters 3 2014-07-01 2014-07-01 false Kennebec River, Bath Iron Works Shipyard, naval restricted area, Bath, Maine. 334.45 Section 334.45 Navigation and Navigable Waters CORPS... REGULATIONS § 334.45 Kennebec River, Bath Iron Works Shipyard, naval restricted area, Bath, Maine. (a)...

  11. 33 CFR 334.45 - Kennebec River, Bath Iron Works Shipyard, naval restricted area, Bath, Maine.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 33 Navigation and Navigable Waters 3 2011-07-01 2011-07-01 false Kennebec River, Bath Iron Works Shipyard, naval restricted area, Bath, Maine. 334.45 Section 334.45 Navigation and Navigable Waters CORPS... REGULATIONS § 334.45 Kennebec River, Bath Iron Works Shipyard, naval restricted area, Bath, Maine. (a)...

  12. 33 CFR 334.45 - Kennebec River, Bath Iron Works Shipyard, naval restricted area, Bath, Maine.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Kennebec River, Bath Iron Works Shipyard, naval restricted area, Bath, Maine. 334.45 Section 334.45 Navigation and Navigable Waters CORPS... REGULATIONS § 334.45 Kennebec River, Bath Iron Works Shipyard, naval restricted area, Bath, Maine. (a)...

  13. Bath-induced paroxysmal disorders in infancy.

    PubMed

    Nechay, Alla; Stephenson, John B P

    2009-05-01

    We reviewed those paroxysmal disorders of infancy and of the newborn in which the normal process of bathing may be an important trigger. We focused on infant bathing in normal temperature water (37 degrees C, range 36-38 degrees C) rather than in hot water that is above body temperature. Four principal diagnostic categories emerged: bathing epilepsy, alternating hemiplegia of childhood, hyperekplexia and paroxysmal extreme pain disorder. Bathing or water immersion epilepsy was the best studied and is arguably distinct from hot water epilepsy. The paroxysmal episodes previously attributed to aquagenic urticaria may have been examples of bathing epilepsy with a genetic component. Despite suggestions in the literature to the contrary, no convincing reports of bath-induced infantile syncope have been found. The underlying mechanisms of bath-induced paroxysmal disorders in infancy remain poorly understood, but all have autonomic manifestations and some if not all may be channelopathies. PMID:18571948

  14. Rhodobacter capsulatus nifA1 Promoter: High-GC −10 Regions in High-GC Bacteria and the Basis for Their Transcription

    PubMed Central

    Richard, Cynthia L.; Tandon, Animesh; Kranz, Robert G.

    2004-01-01

    It was previously shown that the Rhodobacter capsulatus NtrC enhancer-binding protein activates the R. capsulatus housekeeping RNA polymerase but not the Escherichia coli RNA polymerase at the nifA1 promoter. We have tested the hypothesis that this activity is due to the high G+C content of the −10 sequence. A comparative analysis of R. capsulatus and other α-proteobacterial promoters with known transcription start sites suggests that the G+C content of the −10 region is higher than that for E. coli. Both in vivo and in vitro results obtained with nifA1 promoters with −10 and/or −35 variations are reported here. A major conclusion of this study is that α-proteobacteria have evolved a promiscuous sigma factor and core RNA polymerase that can transcribe promoters with high-GC −10 regions in addition to the classic E. coli Pribnow box. To facilitate studies of R. capsulatus transcription, we cloned and overexpressed all of the RNA polymerase subunits in E. coli, and these were reconstituted in vitro to form an active, recombinant R. capsulatus RNA polymerase with properties mimicking those of the natural polymerase. Thus, no additional factors from R. capsulatus are necessary for the recognition of high-GC promoters or for activation by R. capsulatus NtrC. The addition of R. capsulatus σ70 to the E. coli core RNA polymerase or the use of −10 promoter mutants did not facilitate R. capsulatus NtrC activation of the nifA1 promoter by the E. coli RNA polymerase. Thus, an additional barrier to activation by R. capsulatus NtrC exists, probably a lack of the proper R. capsulatus NtrC-E. coli RNA polymerase (protein-protein) interaction(s). PMID:14729700

  15. Bed bathing patients in hospital.

    PubMed

    Downey, Lindsey; Lloyd, Hilary

    There are a number of circumstances that may affect an individual's ability to maintain personal hygiene. Hospitalised patients, and in particular those who are bedridden, may become dependent on nursing staff to carry out their hygiene needs. Assisting patients to maintain personal hygiene is a fundamental aspect of nursing care. However, it is a task often delegated to junior or newly qualified staff. This article focuses on the principles of bed bathing patients in hospital, correct procedure and the importance of maintaining patient dignity and respect in clinical practice. PMID:18543852

  16. [Ofuji papuloerythroderma: PUVA bath treatment].

    PubMed

    Michel, S; Hohenleutner, U; Landthaler, M

    1999-05-01

    Papuloerythroderma of Ofuji is a rare skin disorder described primarily in Japanese patients. It occurs primarily in elderly men. The initial lesions are diffuse red papules, sparing the face, palms and soles. Later the papules coalesce into an erythroderma, with typical sparing of the skin folds and creases (the deck chair sign). Pruritus is usually intense. Lymphadenopathy, peripheral blood eosinophilia and elevated IgE levels all are common. Both systemic corticosteroids and systemic PUVA therapy have been recommended. We describe a German male who fulfilled the diagnostic criteria for papuloerythroderma of Ofuji and responded well to PUVA bath therapy with both improvement in skin findings and reduction in pruritus. PMID:10412634

  17. Extracellular production of tellurium nanoparticles by the photosynthetic bacterium Rhodobacter capsulatus.

    PubMed

    Borghese, Roberto; Brucale, Marco; Fortunato, Gianuario; Lanzi, Massimiliano; Mezzi, Alessio; Valle, Francesco; Cavallini, Massimiliano; Zannoni, Davide

    2016-05-15

    The toxic oxyanion tellurite (TeO3(2-)) is acquired by cells of Rhodobacter capsulatus grown anaerobically in the light, via acetate permease ActP2 and then reduced to Te(0) in the cytoplasm as needle-like black precipitates. Interestingly, photosynthetic cultures of R. capsulatus can also generate Te(0) nanoprecipitates (TeNPs) outside the cells upon addition of the redox mediator lawsone (2-hydroxy-1,4-naphtoquinone). TeNPs generation kinetics were monitored to define the optimal conditions to produce TeNPs as a function of various carbon sources and lawsone concentration. We report that growing cultures over a 10 days period with daily additions of 1mM tellurite led to the accumulation in the growth medium of TeNPs with dimensions from 200 up to 600-700 nm in length as determined by atomic force microscopy (AFM). This result suggests that nucleation of TeNPs takes place over the entire cell growth period although the addition of new tellurium Te(0) to pre-formed TeNPs is the main strategy used by R. capsulatus to generate TeNPs outside the cells. Finally, X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FT-IR) analysis of TeNPs indicate they are coated with an organic material which keeps the particles in solution in aqueous solvents. PMID:26894294

  18. Transcriptional Profiling of Hydrogen Production Metabolism of Rhodobacter capsulatus under Temperature Stress by Microarray Analysis.

    PubMed

    Gürgan, Muazzez; Erkal, Nilüfer Afşar; Özgür, Ebru; Gündüz, Ufuk; Eroglu, Inci; Yücel, Meral

    2015-01-01

    Biohydrogen is a clean and renewable form of hydrogen, which can be produced by photosynthetic bacteria in outdoor large-scale photobioreactors using sunlight. In this study, the transcriptional response of Rhodobacter capsulatus to cold (4 °C) and heat (42 °C) stress was studied using microarrays. Bacteria were grown in 30/2 acetate/glutamate medium at 30 °C for 48 h under continuous illumination. Then, cold and heat stresses were applied for two and six hours. Growth and hydrogen production were impaired under both stress conditions. Microarray chips for R. capsulatus were custom designed by Affymetrix (GeneChip®. TR_RCH2a520699F). The numbers of significantly changed genes were 328 and 293 out of 3685 genes under cold and heat stress, respectively. Our results indicate that temperature stress greatly affects the hydrogen production metabolisms of R. capsulatus. Specifically, the expression of genes that participate in nitrogen metabolism, photosynthesis and the electron transport system were induced by cold stress, while decreased by heat stress. Heat stress also resulted in down regulation of genes related to cell envelope, transporter and binding proteins. Transcriptome analysis and physiological results were consistent with each other. The results presented here may aid clarification of the genetic mechanisms for hydrogen production in purple non-sulfur (PNS) bacteria under temperature stress. PMID:26086826

  19. Transcriptional Profiling of Hydrogen Production Metabolism of Rhodobacter capsulatus under Temperature Stress by Microarray Analysis

    PubMed Central

    Gürgan, Muazzez; Afşar Erkal, Nilüfer; Özgür, Ebru; Gündüz, Ufuk; Eroglu, Inci; Yücel, Meral

    2015-01-01

    Biohydrogen is a clean and renewable form of hydrogen, which can be produced by photosynthetic bacteria in outdoor large-scale photobioreactors using sunlight. In this study, the transcriptional response of Rhodobacter capsulatus to cold (4 °C) and heat (42 °C) stress was studied using microarrays. Bacteria were grown in 30/2 acetate/glutamate medium at 30 °C for 48 h under continuous illumination. Then, cold and heat stresses were applied for two and six hours. Growth and hydrogen production were impaired under both stress conditions. Microarray chips for R. capsulatus were custom designed by Affymetrix (GeneChip®. TR_RCH2a520699F). The numbers of significantly changed genes were 328 and 293 out of 3685 genes under cold and heat stress, respectively. Our results indicate that temperature stress greatly affects the hydrogen production metabolisms of R. capsulatus. Specifically, the expression of genes that participate in nitrogen metabolism, photosynthesis and the electron transport system were induced by cold stress, while decreased by heat stress. Heat stress also resulted in down regulation of genes related to cell envelope, transporter and binding proteins. Transcriptome analysis and physiological results were consistent with each other. The results presented here may aid clarification of the genetic mechanisms for hydrogen production in purple non-sulfur (PNS) bacteria under temperature stress. PMID:26086826

  20. Draft Genome Sequences of Two Heat-Resistant Mutant Strains (A52 and B41) of the Photosynthetic Hydrogen-Producing Bacterium Rhodobacter capsulatus.

    PubMed

    Gokce, Abdulmecit; Cakar, Zeynep Petek; Yucel, Meral; Ozcan, Orhan; Sencan, Sevde; Sertdemir, Ibrahim; Erguner, Bekir; Yuceturk, Betul; Sarac, Aydan; Yuksel, Bayram; Ozturk, Yavuz

    2016-01-01

    The draft genome sequences of two heat-resistant mutant strains, A52 and B41, derived from Rhodobacter capsulatus DSM 1710, and with different hydrogen production levels, are reported here. These sequences may help understand the molecular basis of heat resistance and hydrogen production in R. capsulatus. PMID:27284151

  1. Draft Genome Sequences of Two Heat-Resistant Mutant Strains (A52 and B41) of the Photosynthetic Hydrogen-Producing Bacterium Rhodobacter capsulatus

    PubMed Central

    Gokce, Abdulmecit; Cakar, Zeynep Petek; Yucel, Meral; Ozcan, Orhan; Sencan, Sevde; Sertdemir, Ibrahim; Erguner, Bekir; Yuceturk, Betul; Sarac, Aydan; Yuksel, Bayram

    2016-01-01

    The draft genome sequences of two heat-resistant mutant strains, A52 and B41, derived from Rhodobacter capsulatus DSM 1710, and with different hydrogen production levels, are reported here. These sequences may help understand the molecular basis of heat resistance and hydrogen production in R. capsulatus. PMID:27284151

  2. Noncyanide cadmium plating baths. Final report

    SciTech Connect

    Pearlstein, F.; Agarwala, V.S.

    1991-10-04

    One approach to minimizing toxic wastes is to eliminate the use of cyanide plating baths. Non-cyanide zinc plating baths have been successfully developed and have found widespread use. An investigation was conducted in an attempt to accomplish similar results with cadmium plating baths. The focus of this study was on additives to a near neutral cadmium bath, free of complexing agents. A Hull cell was used to enable visualization of deposits over a broad range of cathode current densities. Experimental design (Taguchi Method) was used to optimize bath parameters and constituent concentrations. Bath parameters have been developed which indicate promise for producing dense deposits with good covering power, and relatively low tendency for hydrogen embrittlement.

  3. Bathing Epilepsy: Report of Three Caucasian Cases

    PubMed Central

    Dashi, Florian; Seferi, Arsen; Rroji, Arben; Enesi, Eugen; Petrela, Mentor

    2015-01-01

    Introduction: Bathing epilepsy is a specific type of reflex epilepsy triggered by domestic bathing in water. It is a geographically specific epilepsy syndrome that is more prevalent in India Cases in Caucasian population are very rarely reported. These cases share many similar clinical features and a similar prognosis to the Indian cases. Case report: We describe three cases of bathing epilepsy in Albanian population; two cases with well controlled seizures and one with drug-resistant seizures. PMID:26005279

  4. Mobile wedges in an active turbulent bath

    NASA Astrophysics Data System (ADS)

    Kaiser, Andreas; Sokolov, Andrey; Lowen, Hartmut; Aronson, Igor S.

    The motion of micro-wedges in a turbulent bacterial bath is explored using computer simulations with explicit modeling of the bacteria and experiments. We demonstrate that collective turbulentlike motion in a bacterial bath can power and steer the directed transport of mesoscopic carriers through the suspension. We will show that both polar ordering and swirl shielding inside the wedge yield an optimal transport velocity. Finally, we show the behavior of several wedges exposed to a bacterial bath.

  5. Finite-Size Bath in Qubit Thermodynamics

    NASA Astrophysics Data System (ADS)

    Pekola, J. P.; Suomela, S.; Galperin, Y. M.

    2016-09-01

    We discuss a qubit weakly coupled to a finite-size heat bath (calorimeter) from the point of view of quantum thermodynamics. The energy deposited to this environment together with the state of the qubit provides a basis to analyze the heat and work statistics of this closed combined system. We present results on two representative models, where the bath is composed of two-level systems or harmonic oscillators, respectively. Finally, we derive results for an open quantum system composed of the above qubit plus finite-size bath, but now the latter is coupled to a practically infinite bath of the same nature of oscillators or two-level systems.

  6. Finite-Size Bath in Qubit Thermodynamics

    NASA Astrophysics Data System (ADS)

    Pekola, J. P.; Suomela, S.; Galperin, Y. M.

    2016-04-01

    We discuss a qubit weakly coupled to a finite-size heat bath (calorimeter) from the point of view of quantum thermodynamics. The energy deposited to this environment together with the state of the qubit provides a basis to analyze the heat and work statistics of this closed combined system. We present results on two representative models, where the bath is composed of two-level systems or harmonic oscillators, respectively. Finally, we derive results for an open quantum system composed of the above qubit plus finite-size bath, but now the latter is coupled to a practically infinite bath of the same nature of oscillators or two-level systems.

  7. Bathing or washing babies after birth?

    PubMed

    Henningsson, A; Nyström, B; Tunnell, R

    One group of healthy full-term newborn babies was washed after birth and another was bathed to remove vernix caseosa and clean the skin. Few infections, none of them serious, occurred in either group. Bacterial colonisation of the umbilical cord on the third day of life was similar in both groups. The rectal temperature fell further and more infants cried during washing than during bathing. Thus bathing the baby after birth makes it calmer, quieter, and more comfortable than washing and causes less heat-loss. Clinical signs of infection and bacterial colonisation rates are no higher after bathing than after washing. PMID:6118769

  8. Pulling bubbles from a bath

    NASA Astrophysics Data System (ADS)

    Kao, Justin C. T.; Blakemore, Andrea L.; Hosoi, A. E.

    2010-06-01

    Deposition of bubbles on a wall withdrawn from a liquid bath is a phenomenon observed in many everyday situations—the foam lacing left behind in an emptied glass of beer, for instance. It is also of importance to the many industrial processes where uniformity of coating is desirable. We report work on an idealized version of this situation, the drag-out of a single bubble in Landau-Levich-Derjaguin flow. We find that a well-defined critical wall speed exists, separating the two regimes of bubble persistence at the meniscus and bubble deposition on the moving wall. Experiments show that this transition occurs at Ca∗˜Bo0.73. A similar result is obtained theoretically by balancing viscous stresses and gravity.

  9. 21 CFR 890.5110 - Paraffin bath.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Paraffin bath. 890.5110 Section 890.5110 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES PHYSICAL MEDICINE DEVICES Physical Medicine Therapeutic Devices § 890.5110 Paraffin bath....

  10. 21 CFR 890.5110 - Paraffin bath.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Paraffin bath. 890.5110 Section 890.5110 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES PHYSICAL MEDICINE DEVICES Physical Medicine Therapeutic Devices § 890.5110 Paraffin bath....

  11. 21 CFR 890.5110 - Paraffin bath.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Paraffin bath. 890.5110 Section 890.5110 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES PHYSICAL MEDICINE DEVICES Physical Medicine Therapeutic Devices § 890.5110 Paraffin bath....

  12. 21 CFR 890.5110 - Paraffin bath.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Paraffin bath. 890.5110 Section 890.5110 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES PHYSICAL MEDICINE DEVICES Physical Medicine Therapeutic Devices § 890.5110 Paraffin bath....

  13. Temperature control of a cryogenic bath

    NASA Technical Reports Server (NTRS)

    Asher, I. M.

    1972-01-01

    Foreign gas introduced into vapor phase above liquid region cools cryogenic baths. Equipment consists of gas tank and cover of styrofoam. Helium is considered the best choice to produce cooling, though any gas with boiling point lower than that of bath liquid may be used.

  14. 21 CFR 890.5110 - Paraffin bath.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Paraffin bath. 890.5110 Section 890.5110 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES PHYSICAL MEDICINE DEVICES Physical Medicine Therapeutic Devices § 890.5110 Paraffin bath....

  15. 7 CFR 3201.62 - Bath products.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 15 2014-01-01 2014-01-01 false Bath products. 3201.62 Section 3201.62 Agriculture Regulations of the Department of Agriculture (Continued) OFFICE OF PROCUREMENT AND PROPERTY MANAGEMENT, DEPARTMENT OF AGRICULTURE GUIDELINES FOR DESIGNATING BIOBASED PRODUCTS FOR FEDERAL PROCUREMENT Designated Items § 3201.62 Bath products....

  16. 7 CFR 3201.62 - Bath products.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 15 2012-01-01 2012-01-01 false Bath products. 3201.62 Section 3201.62 Agriculture Regulations of the Department of Agriculture (Continued) OFFICE OF PROCUREMENT AND PROPERTY MANAGEMENT, DEPARTMENT OF AGRICULTURE GUIDELINES FOR DESIGNATING BIOBASED PRODUCTS FOR FEDERAL PROCUREMENT Designated Items § 3201.62 Bath products....

  17. 7 CFR 3201.62 - Bath products.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 15 2013-01-01 2013-01-01 false Bath products. 3201.62 Section 3201.62 Agriculture Regulations of the Department of Agriculture (Continued) OFFICE OF PROCUREMENT AND PROPERTY MANAGEMENT, DEPARTMENT OF AGRICULTURE GUIDELINES FOR DESIGNATING BIOBASED PRODUCTS FOR FEDERAL PROCUREMENT Designated Items § 3201.62 Bath products....

  18. New system for bathing bedridden patients

    NASA Technical Reports Server (NTRS)

    Greenleaf, J. E.; Staley, R. A.; Payne, P. A.

    1973-01-01

    Multihead shower facility can be used with minimal patient handling. Waterproof curtain allows patient to bathe with his head out of shower. He can move completely inside shower to wash his face and hair. Main advantage of shower system is time saved in giving bath.

  19. Alternatives to hexachlorophene bathing of newborn infants.

    PubMed Central

    Hnatko, S. I.

    1977-01-01

    In controlled trials newborn infants were bathed with Lactacyd, pHisoHex, Hibitane, Lanohex or tap water. Bacteriologic samples were taken from three sites (groin, axilla and cord) immediately after birth, following an initial bath with one of the test agents, and on day 3 or 5 after a water bath. Initial bathing with all agents, including water, reduced the concentration of bacteria on the skin to a similar extent. However, comparisons of bacterial flora at birth versus those on days 3 and 5 indicated differences in the actions of the various agents on pathogenic and nonpathogenic organisms. Lactacyd and Hibitane appeared to be suitable alternatives to hexachlorophene in the control of pathogenic bacteria on the skin of newborns. However, their absorption and toxicity in the newborn are unknown and, unless use of a skin disinfectant is warranted, routine bathing of newborns with tap water appears to be satisfactory. PMID:328126

  20. Methylogaea oryzae gen. nov., sp. nov., a mesophilic methanotroph isolated from a rice paddy field.

    PubMed

    Geymonat, Estefanía; Ferrando, Lucía; Tarlera, Silvana E

    2011-11-01

    A novel methanotroph, designated strain E10(T), was isolated from a rice paddy field in Uruguay. Strain E10(T) grew on methane and methanol as sole carbon and energy sources. Cells were Gram-negative, non-motile, non-pigmented, slightly curved rods showing type I intracytoplasmic membranes arranged in stacks. The strain was neutrophilic and mesophilic; optimum growth occurred at 30-35 °C with no growth above 37 °C. The strain possessed only a particulate methane monooxygenase (pmoA). Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strain was most closely related to the moderately thermophilic strains Methylocaldum szegediense OR2(T) (91.6 % sequence similarity) and Methylococcus capsulatus Bath (91.5 %). Comparative sequence analysis of pmoA genes also confirmed that strain E10(T) formed a new lineage among the genera Methylocaldum and Methylococcus with 89 and 84 % derived amino acid sequence identity to Methylococcus capsulatus Bath and Methylocaldum gracile VKM-14L(T), respectively. The DNA G+C content was 63.1 mol% and the major cellular fatty acid was C(16 :0) (62.05 %). Thus, strain E10(T) (=JCM 16910(T) = DSM 23452(T)) represents the type strain of a novel species within a new genus, for which the name Methylogaea oryzae gen. nov., sp. nov. is proposed. PMID:21131502

  1. Dataset of gene cloning and gel filtration chromatography of R-est6.

    PubMed

    Soni, Surabhi; Odaneth, Annamma A; Lali, Arvind M; Chandrayan, Sanjeev K

    2016-06-01

    The data presented in this article are connected to the research article entitled "Expression, purification and biochemical characterization of a family 6 carboxylesterase from Methylococcus capsulatus (bath)" (Soni et al., 2016 [1]). The family 6 carboxylesterases are the smallest and display broad substrate specificity. The 1 kb gene encoding, a family 6 carboxylesterase - R-est6, was amplified from the genome of M. capsulatus (bath strain), and showed in the agarose gel. The corresponding purified protein, after overexpression in Escherichia coli, was biochemically studied in the research article (Soni et al., 2016 [1]). R-est6 has hydrophobic patches on the surface so, it is expected to show oligimeric forms. Here, we have confirmed the presence of oligomers by gel filtration chromatography data and the proteins belonging to the different peaks are shown on a SDS-PAGE. PMID:27222859

  2. Dataset of gene cloning and gel filtration chromatography of R-est6

    PubMed Central

    Soni, Surabhi; Odaneth, Annamma A.; Lali, Arvind M.; Chandrayan, Sanjeev K.

    2016-01-01

    The data presented in this article are connected to the research article entitled “Expression, purification and biochemical characterization of a family 6 carboxylesterase from Methylococcus capsulatus (bath)” (Soni et al., 2016 [1]). The family 6 carboxylesterases are the smallest and display broad substrate specificity. The 1 kb gene encoding, a family 6 carboxylesterase – R-est6, was amplified from the genome of M. capsulatus (bath strain), and showed in the agarose gel. The corresponding purified protein, after overexpression in Escherichia coli, was biochemically studied in the research article (Soni et al., 2016 [1]). R-est6 has hydrophobic patches on the surface so, it is expected to show oligimeric forms. Here, we have confirmed the presence of oligomers by gel filtration chromatography data and the proteins belonging to the different peaks are shown on a SDS-PAGE. PMID:27222859

  3. Mechanism of Substrate and Inhibitor Binding of Rhodobacter Capsulatus Xanthine Dehydrogenase

    SciTech Connect

    Dietzel, U.; Kuper, J; Doebbler, J; Schulte, A; Truglio, J; Leimkuhler, S; Kisker, C

    2009-01-01

    Rhodobacter capsulatus xanthine dehydrogenase (XDH) is an (ae)2 heterotetrameric cytoplasmic enzyme that resembles eukaryotic xanthine oxidoreductases in respect to both amino acid sequence and structural fold. To obtain a detailed understanding of the mechanism of substrate and inhibitor binding at the active site, we solved crystal structures of R. capsulatus XDH in the presence of its substrates hypoxanthine, xanthine, and the inhibitor pterin-6-aldehyde using either the inactive desulfo form of the enzyme or an active site mutant (EB232Q) to prevent substrate turnover. The hypoxanthine- and xanthine-bound structures reveal the orientation of both substrates at the active site and show the importance of residue GluB-232 for substrate positioning. The oxygen atom at the C-6 position of both substrates is oriented toward ArgB-310 in the active site. Thus the substrates bind in an orientation opposite to the one seen in the structure of the reduced enzyme with the inhibitor oxypurinol. The tightness of the substrates in the active site suggests that the intermediate products must exit the binding pocket to allow first the attack of the C-2, followed by oxidation of the C-8 atom to form the final product uric acid. Structural studies of pterin-6-aldehyde, a potent inhibitor of R. capsulatus XDH, contribute further to the understanding of the relative positioning of inhibitors and substrates in the binding pocket. Steady state kinetics reveal a competitive inhibition pattern with a Ki of 103.57 {+-} 18.96 nm for pterin-6-aldehyde.

  4. Cu2+ site in photosynthetic bacterial reaction centers from Rhodobacter sphaeroides, Rhodobacter capsulatus, and Rhodopseudomonas viridis.

    PubMed

    Utschig, L M; Poluektov, O; Schlesselman, S L; Thurnauer, M C; Tiede, D M

    2001-05-22

    The interaction of metal ions with isolated photosynthetic reaction centers (RCs) from the purple bacteria Rhodobacter sphaeroides, Rhodobacter capsulatus, and Rhodopseudomonas viridis has been investigated with transient optical and magnetic resonance techniques. In RCs from all species, the electrochromic response of the bacteriopheophytin cofactors associated with Q(A)(-)Q(B) --> Q(A)Q(B)(-) electron transfer is slowed in the presence of Cu(2+). This slowing is similar to the metal ion effect observed for RCs from Rb. sphaeroides where Zn(2+) was bound to a specific site on the surface of the RC [Utschig et al. (1998) Biochemistry 37, 8278]. The coordination environments of the Cu(2+) sites were probed with electron paramagnetic resonance (EPR) spectroscopy, providing the first direct spectroscopic evidence for the existence of a second metal site in RCs from Rb. capsulatus and Rps. viridis. In the dark, RCs with Cu(2+) bound to the surface exhibit axially symmetric EPR spectra. Electron spin echo envelope modulation (ESEEM) spectral results indicate multiple weakly hyperfine coupled (14)N nuclei in close proximity to Cu(2+). These ESEEM spectra resemble those observed for Cu(2+) RCs from Rb. sphaeroides [Utschig et al. (2000) Biochemistry 39, 2961] and indicate that two or more histidines ligate the Cu(2+) at the surface site in each RC. Thus, RCs from Rb. sphaeroides, Rb. capsulatus, and Rps. viridis each have a structurally analogous Cu(2+) binding site that is involved in modulating the Q(A)(-)Q(B) --> Q(A)Q(B)(-) electron-transfer process. Inspection of the Rps. viridis crystal structure reveals four potential histidine ligands from three different subunits (M16, H178, H72, and L211) located beneath the Q(B) binding pocket. The location of these histidines is surprisingly similar to the grouping of four histidine residues (H68, H126, H128, and L211) observed in the Rb. sphaeroides RC crystal structure. Further elucidation of these Cu(2+) sites will provide

  5. Gene co-expression network analysis in Rhodobacter capsulatus and application to comparative expression analysis of Rhodobacter sphaeroides

    SciTech Connect

    Pena-Castillo, Lourdes; Mercer, Ryan; Gurinovich, Anastasia; Callister, Stephen J.; Wright, Aaron T.; Westbye, Alexander; Beatty, J. T.; Lang, Andrew S.

    2014-08-28

    The genus Rhodobacter contains purple nonsulfur bacteria found mostly in freshwater environments. Representative strains of two Rhodobacter species, R. capsulatus and R. sphaeroides, have had their genomes fully sequenced and both have been the subject of transcriptional profiling studies. Gene co-expression networks can be used to identify modules of genes with similar expression profiles. Functional analysis of gene modules can then associate co-expressed genes with biological pathways, and network statistics can determine the degree of module preservation in related networks. In this paper, we constructed an R. capsulatus gene co-expression network, performed functional analysis of identified gene modules, and investigated preservation of these modules in R. capsulatus proteomics data and in R. sphaeroides transcriptomics data. Results: The analysis identified 40 gene co-expression modules in R. capsulatus. Investigation of the module gene contents and expression profiles revealed patterns that were validated based on previous studies supporting the biological relevance of these modules. We identified two R. capsulatus gene modules preserved in the protein abundance data. We also identified several gene modules preserved between both Rhodobacter species, which indicate that these cellular processes are conserved between the species and are candidates for functional information transfer between species. Many gene modules were non-preserved, providing insight into processes that differentiate the two species. In addition, using Local Network Similarity (LNS), a recently proposed metric for expression divergence, we assessed the expression conservation of between-species pairs of orthologs, and within-species gene-protein expression profiles. Conclusions: Our analyses provide new sources of information for functional annotation in R. capsulatus because uncharacterized genes in modules are now connected with groups of genes that constitute a joint functional

  6. Regulation of synthesis of pyruvate carboxylase in the photosynthetic bacterium Rhodobacter capsulatus.

    PubMed Central

    Yakunin, A F; Hallenbeck, P C

    1997-01-01

    The synthesis of pyruvate carboxylase (PC) was studied by using quantitative immunoblot analysis with an antibody raised against PC purified from Rhodobacter capsulatus and was found to vary 20-fold depending on the growth conditions. The PC content was high in cells grown on pyruvate or on carbon substrates metabolized via pyruvate (lactate, D-malate, glucose, or fructose) and low in cells grown on tricarboxylic acid (TCA) cycle intermediates or substrates metabolized without intermediate formation of pyruvate (acetate or glutamate). Under dark aerobic growth conditions with lactate as a carbon source, the PC content was approximately twofold higher than that found under light anaerobic growth conditions. The results of incubation experiments demonstrate that PC synthesis is induced by pyruvate and repressed by TCA cycle intermediates, with negative control dominating over positive control. The content of PC in R. capsulatus cells was also directly related to the growth rate in continuous cultures. The analysis of intracellular levels of pyruvate and TCA cycle intermediates in cells grown under different conditions demonstrated that the content of PC is directly proportional to the ratio between pyruvate and C4 dicarboxylates. These results suggest that the regulation of PC synthesis by oxygen and its direct correlation with growth rate may reflect effects on the balance of intracellular pyruvate and C4 dicarboxylates. Thus, this important enzyme is potentially regulated both allosterically and at the level of synthesis. PMID:9045800

  7. Isolation of mutants and genes involved in cytochromes c biosynthesis in Rhodobacter capsulatus.

    PubMed Central

    Kranz, R G

    1989-01-01

    Mutants of the photosynthetic bacterium Rhodobacter capsulatus that have combined deficiencies in the cytochrome b/c1 complex and other c-type cytochromes have been isolated. These mutants were unable to grow anaerobically in the light or dark but could grow aerobically. Cosmids with R. capsulatus wild-type DNA that complement the mutants have been used to construct genetic and physical maps of the affected genes. Complementation profiles with Tn5 and mini-Mu insertions in these cosmids and subcloned fragments from them indicated that at least three genes (called helA, helB, and helC) are involved in the defects in cytochromes c biosynthesis. The genes are clustered, and helC is transcribed away from helA and helB. Stable insertion mutants in each gene were constructed. It is postulated that helA, helB, and helC are involved in posttranslational processing during cytochromes c synthesis. Images PMID:2536664

  8. A Rhodobacter capsulatus Member of a Universal Permease Family Imports Molybdate and Other Oxyanions▿

    PubMed Central

    Gisin, Jonathan; Müller, Alexandra; Pfänder, Yvonne; Leimkühler, Silke; Narberhaus, Franz; Masepohl, Bernd

    2010-01-01

    Molybdenum (Mo) is an important trace element that is toxic at high concentrations. To resolve the mechanisms underlying Mo toxicity, Rhodobacter capsulatus mutants tolerant to high Mo concentrations were isolated by random transposon Tn5 mutagenesis. The insertion sites of six independent isolates mapped within the same gene predicted to code for a permease of unknown function located in the cytoplasmic membrane. During growth under Mo-replete conditions, the wild-type strain accumulated considerably more Mo than the permease mutant. For mutants defective for the permease, the high-affinity molybdate importer ModABC, or both transporters, in vivo Mo-dependent nitrogenase (Mo-nitrogenase) activities at different Mo concentrations suggested that ModABC and the permease import molybdate in nanomolar and micromolar ranges, respectively. Like the permease mutants, a mutant defective for ATP sulfurylase tolerated high Mo concentrations, suggesting that ATP sulfurylase is the main target of Mo inhibition in R. capsulatus. Sulfate-dependent growth of a double mutant defective for the permease and the high-affinity sulfate importer CysTWA was reduced compared to those of the single mutants, implying that the permease plays an important role in sulfate uptake. In addition, permease mutants tolerated higher tungstate and vanadate concentrations than the wild type, suggesting that the permease acts as a general oxyanion importer. We propose to call this permease PerO (for oxyanion permease). It is the first reported bacterial molybdate transporter outside the ABC transporter family. PMID:20851900

  9. Effects of room temperature on physiological and subjective responses during whole-body bathing, half-body bathing and showering.

    PubMed

    Hashiguchi, Nobuko; Ni, Furong; Tochihara, Yutaka

    2002-11-01

    The effects of bathroom thermal conditions on physiological and subjective responses were evaluated before, during, and after whole-body bath (W-bath), half-body bath (H-bath) and showering. The air temperature of the dressing room and bathroom was controlled at 10 degrees C, 17.5 degrees C, and 25 degrees C. Eight healthy males bathed for 10 min under nine conditions on separate days. The water temperature of the bathtub and shower was controlled at 40 degrees C and 41 degrees C, respectively. Rectal temperature (Tre), mean skin temperature (Tsk), blood pressure (BP), heart rate (HR), body weight loss and blood characteristics (hematocrit: Hct, hemoglobin: Hb) were evaluated. Also, thermal sensation (TS), thermal comfort (TC) and thermal acceptability (TA) were recorded. BP decreased rapidly during W-bath and H-bath compared to showering. HR during W-bath was significantly higher than for H-bath and showering (p < 0.01). The double products due to W-bath during bathing were also greater than for H-bath and showering (p < 0.05). There were no distinct differences in Hct and Hb among the nine conditions. However, significant differences in body weight loss were observed among the bathing methods: W-bath > H-bath > showering (p < 0.001). W-bath showed the largest increase in Tre and Tsk, followed by H-bath, and showering. Significant differences in Tre after bathing among the room temperatures were found only at H-bath. The changes in Tre after bathing for H-bath at 25 degrees C were similar to those for W-bath at 17.5 degrees C and 10 degrees C. TS and TC after bathing significantly differed for the three bathing methods at 17.5 degrees C and 10 degrees C (TS: p < 0.01 TC: p < 0.001). Especially, for showering, the largest number of subjects felt "cold" and "uncomfortable". Even though all of the subjects could accept the 10 degrees C condition after W-bath, such conditions were intolerable to half of them after showering. These results suggested that the

  10. Loss of the Response Regulator CtrA Causes Pleiotropic Effects on Gene Expression but Does Not Affect Growth Phase Regulation in Rhodobacter capsulatus

    SciTech Connect

    Mercer, Ryan; Callister, Stephen J.; Lipton, Mary S.; Pasa-Tolic, Ljiljana; Strnad, Hynek; Paces, Vaclav; Beatty, J. T.; Lang, Andrew S.

    2010-06-01

    The purple non-sulfur bacterium Rhodobacter capsulatus has been extensively studied for its diverse metabolic capabilities, as well as for its production of a Gene Transfer Agent (RcGTA). Production of RcGTA requires the response regulator protein CtrA. We have used whole genome transcript and whole cell proteome analyses of wild type and ctrA mutant cultures to completely characterize the regulatory role of CtrA in R. capsulatus.

  11. Structural and mechanistic insights into methane oxidation by particulate methane monooxygenase.

    PubMed

    Balasubramanian, Ramakrishnan; Rosenzweig, Amy C

    2007-07-01

    Particulate methane monooxygense (pMMO) is an integral membrane copper-containing enzyme that converts methane to methanol. Knowledge of how pMMO selectively oxidizes methane under ambient conditions could impact the development of new catalysts. The crystal structure of Methylococcus capsulatus (Bath) pMMO reveals the composition and location of three metal centers. Spectroscopic data provide insight into the coordination environments and oxidation states of these metal centers. These results, combined with computational studies and comparisons to relevant systems, are discussed in the context of identifying the most likely site for O 2 activation. PMID:17444606

  12. The metal centres of particulate methane mono-oxygenase.

    PubMed

    Rosenzweig, Amy C

    2008-12-01

    pMMO (particulate methane mono-oxygenase) is an integral membrane metalloenzyme that catalyses the oxidation of methane to methanol. The pMMO metal active site has not been identified, precluding detailed investigation of the reaction mechanism. Models for the metal centres proposed by various research groups have evolved as crystallographic and spectroscopic data have become available. The present review traces the evolution of these active-site models before and after the 2005 Methylococcus capsulatus (Bath) pMMO crystal structure determination. PMID:19021511

  13. The Metal Centers of Particulate Methane Monooxygenase from Methylosinus trichosporium OB3b

    SciTech Connect

    Hakemian,A.; Kondapalli, K.; Telser, J.; Hoffman, B.; Stemmler, T.; Rosenzweig, A.

    2008-01-01

    Particulate methane monooxygenase (pMMO) is a membrane-bound metalloenzyme that oxidizes methane to methanol in methanotrophic bacteria. The nature of the pMMO active site and the overall metal content are controversial, with spectroscopic and crystallographic data suggesting the presence of a mononuclear copper center, a dinuclear copper center, a trinuclear center, and a diiron center or combinations thereof. Most studies have focused on pMMO from Methylococcus capsulatus (Bath). pMMO from a second organism, Methylosinus trichosporium OB3b, has been purified and characterized by spectroscopic and crystallographic methods. Purified M. trichosporium OB3b pMMO contains 2 copper ions per 100 kDa protomer. Electron paramagnetic resonance (EPR) spectroscopic parameters indicate that type 2 Cu(II) is present as two distinct species. Extended X-ray absorption fine structure (EXAFS) data are best fit with oxygen/nitrogen ligands and reveal a Cu-Cu interaction at 2.52 Angstroms. Correspondingly, X-ray crystallography of M. trichosporium OB3b pMMO shows a dinuclear copper center, similar to that observed previously in the crystal structure of M. capsulatus (Bath) pMMO. There are, however, significant differences between the pMMO structures from the two organisms. A mononuclear copper center present in M. capsulatus (Bath) pMMO is absent in M. trichosporium OB3b pMMO, whereas a metal center occupied by zinc in the M. capsulatus (Bath) pMMO structure is occupied by copper in M. trichosporium OB3b pMMO. These findings extend previous work on pMMO from M. capsulatus (Bath) and provide new insight into the functional importance of the different metal centers.

  14. The metal centers of particulate methane monooxygenase from Methylosinus trichosporium OB3b.

    PubMed

    Hakemian, Amanda S; Kondapalli, Kalyan C; Telser, Joshua; Hoffman, Brian M; Stemmler, Timothy L; Rosenzweig, Amy C

    2008-07-01

    Particulate methane monooxygenase (pMMO) is a membrane-bound metalloenzyme that oxidizes methane to methanol in methanotrophic bacteria. The nature of the pMMO active site and the overall metal content are controversial, with spectroscopic and crystallographic data suggesting the presence of a mononuclear copper center, a dinuclear copper center, a trinuclear center, and a diiron center or combinations thereof. Most studies have focused on pMMO from Methylococcus capsulatus (Bath). pMMO from a second organism, Methylosinus trichosporium OB3b, has been purified and characterized by spectroscopic and crystallographic methods. Purified M. trichosporium OB3b pMMO contains approximately 2 copper ions per 100 kDa protomer. Electron paramagnetic resonance (EPR) spectroscopic parameters indicate that type 2 Cu(II) is present as two distinct species. Extended X-ray absorption fine structure (EXAFS) data are best fit with oxygen/nitrogen ligands and reveal a Cu-Cu interaction at 2.52 A. Correspondingly, X-ray crystallography of M. trichosporium OB3b pMMO shows a dinuclear copper center, similar to that observed previously in the crystal structure of M. capsulatus (Bath) pMMO. There are, however, significant differences between the pMMO structures from the two organisms. A mononuclear copper center present in M. capsulatus (Bath) pMMO is absent in M. trichosporium OB3b pMMO, whereas a metal center occupied by zinc in the M. capsulatus (Bath) pMMO structure is occupied by copper in M. trichosporium OB3b pMMO. These findings extend previous work on pMMO from M. capsulatus (Bath) and provide new insight into the functional importance of the different metal centers. PMID:18540635

  15. Avian Assemblages at Bird Baths: A Comparison of Urban and Rural Bird Baths in Australia.

    PubMed

    Cleary, Gráinne P; Parsons, Holly; Davis, Adrian; Coleman, Bill R; Jones, Darryl N; Miller, Kelly K; Weston, Michael A

    2016-01-01

    Private gardens provide habitat and resources for many birds living in human-dominated landscapes. While wild bird feeding is recognised as one of the most popular forms of human-wildlife interaction, almost nothing is known about the use of bird baths. This citizen science initiative explores avian assemblages at bird baths in private gardens in south-eastern Australia and how this differs with respect to levels of urbanisation and bioregion. Overall, 992 citizen scientists collected data over two, four-week survey periods during winter 2014 and summer 2015 (43% participated in both years). Avian assemblages at urban and rural bird baths differed between bioregions with aggressive nectar-eating species influenced the avian assemblages visiting urban bird baths in South Eastern Queensland, NSW North Coast and Sydney Basin while introduced birds contributed to differences in South Western Slopes, Southern Volcanic Plains and Victorian Midlands. Small honeyeaters and other small native birds occurred less often at urban bird baths compared to rural bird baths. Our results suggest that differences between urban versus rural areas, as well as bioregion, significantly influence the composition of avian assemblages visiting bird baths in private gardens. We also demonstrate that citizen science monitoring of fixed survey sites such as bird baths is a useful tool in understanding large-scale patterns in avian assemblages which requires a vast amount of data to be collected across broad areas. PMID:26962857

  16. Avian Assemblages at Bird Baths: A Comparison of Urban and Rural Bird Baths in Australia

    PubMed Central

    Cleary, Gráinne P.; Parsons, Holly; Davis, Adrian; Coleman, Bill R.; Jones, Darryl N.; Miller, Kelly K.; Weston, Michael A.

    2016-01-01

    Private gardens provide habitat and resources for many birds living in human-dominated landscapes. While wild bird feeding is recognised as one of the most popular forms of human-wildlife interaction, almost nothing is known about the use of bird baths. This citizen science initiative explores avian assemblages at bird baths in private gardens in south-eastern Australia and how this differs with respect to levels of urbanisation and bioregion. Overall, 992 citizen scientists collected data over two, four-week survey periods during winter 2014 and summer 2015 (43% participated in both years). Avian assemblages at urban and rural bird baths differed between bioregions with aggressive nectar-eating species influenced the avian assemblages visiting urban bird baths in South Eastern Queensland, NSW North Coast and Sydney Basin while introduced birds contributed to differences in South Western Slopes, Southern Volcanic Plains and Victorian Midlands. Small honeyeaters and other small native birds occurred less often at urban bird baths compared to rural bird baths. Our results suggest that differences between urban versus rural areas, as well as bioregion, significantly influence the composition of avian assemblages visiting bird baths in private gardens. We also demonstrate that citizen science monitoring of fixed survey sites such as bird baths is a useful tool in understanding large-scale patterns in avian assemblages which requires a vast amount of data to be collected across broad areas. PMID:26962857

  17. Rhodobacter capsulatus OlsA is a bifunctional enzyme active in both ornithine lipid and phosphatidic acid biosynthesis.

    PubMed

    Aygun-Sunar, Semra; Bilaloglu, Rahmi; Goldfine, Howard; Daldal, Fevzi

    2007-12-01

    The Rhodobacter capsulatus genome contains three genes (olsA [plsC138], plsC316, and plsC3498) that are annotated as lysophosphatidic acid (1-acyl-sn-glycerol-3-phosphate) acyltransferase (AGPAT). Of these genes, olsA was previously shown to be an O-acyltransferase in the second step of ornithine lipid biosynthesis, which is important for optimal steady-state levels of c-type cytochromes (S. Aygun-Sunar, S. Mandaci, H.-G. Koch, I. V. J. Murray, H. Goldfine, and F. Daldal. Mol. Microbiol. 61:418-435, 2006). The roles of the remaining plsC316 and plsC3498 genes remained unknown. In this work, these genes were cloned, and chromosomal insertion-deletion mutations inactivating them were obtained to define their function. Characterization of these mutants indicated that, unlike the Escherichia coli plsC, neither plsC316 nor plsC3498 was essential in R. capsulatus. In contrast, no plsC316 olsA double mutant could be isolated, indicating that an intact copy of either olsA or plsC316 was required for R. capsulatus growth under the conditions tested. Compared to OlsA null mutants, PlsC316 null mutants contained ornithine lipid and had no c-type cytochrome-related phenotype. However, they exhibited slight growth impairment and highly altered total fatty acid and phospholipid profiles. Heterologous expression in an E. coli plsC(Ts) mutant of either R. capsulatus plsC316 or olsA gene products supported growth at a nonpermissive temperature, exhibited AGPAT activity in vitro, and restored phosphatidic acid biosynthesis. The more vigorous AGPAT activity displayed by PlsC316 suggested that plsC316 encodes the main AGPAT required for glycerophospholipid synthesis in R. capsulatus, while olsA acts as an alternative AGPAT that is specific for ornithine lipid synthesis. This study therefore revealed for the first time that some OlsA enzymes, like the enzyme of R. capsulatus, are bifunctional and involved in both membrane ornithine lipid and glycerophospholipid biosynthesis. PMID

  18. Rhodobacter capsulatus OlsA Is a Bifunctional Enyzme Active in both Ornithine Lipid and Phosphatidic Acid Biosynthesis▿ †

    PubMed Central

    Aygun-Sunar, Semra; Bilaloglu, Rahmi; Goldfine, Howard; Daldal, Fevzi

    2007-01-01

    The Rhodobacter capsulatus genome contains three genes (olsA [plsC138], plsC316, and plsC3498) that are annotated as lysophosphatidic acid (1-acyl-sn-glycerol-3-phosphate) acyltransferase (AGPAT). Of these genes, olsA was previously shown to be an O-acyltransferase in the second step of ornithine lipid biosynthesis, which is important for optimal steady-state levels of c-type cytochromes (S. Aygun-Sunar, S. Mandaci, H.-G. Koch, I. V. J. Murray, H. Goldfine, and F. Daldal. Mol. Microbiol. 61:418-435, 2006). The roles of the remaining plsC316 and plsC3498 genes remained unknown. In this work, these genes were cloned, and chromosomal insertion-deletion mutations inactivating them were obtained to define their function. Characterization of these mutants indicated that, unlike the Escherichia coli plsC, neither plsC316 nor plsC3498 was essential in R. capsulatus. In contrast, no plsC316 olsA double mutant could be isolated, indicating that an intact copy of either olsA or plsC316 was required for R. capsulatus growth under the conditions tested. Compared to OlsA null mutants, PlsC316 null mutants contained ornithine lipid and had no c-type cytochrome-related phenotype. However, they exhibited slight growth impairment and highly altered total fatty acid and phospholipid profiles. Heterologous expression in an E. coli plsC(Ts) mutant of either R. capsulatus plsC316 or olsA gene products supported growth at a nonpermissive temperature, exhibited AGPAT activity in vitro, and restored phosphatidic acid biosynthesis. The more vigorous AGPAT activity displayed by PlsC316 suggested that plsC316 encodes the main AGPAT required for glycerophospholipid synthesis in R. capsulatus, while olsA acts as an alternative AGPAT that is specific for ornithine lipid synthesis. This study therefore revealed for the first time that some OlsA enzymes, like the enzyme of R. capsulatus, are bifunctional and involved in both membrane ornithine lipid and glycerophospholipid biosynthesis. PMID

  19. Chlorhexidine: Patient Bathing and Infection Prevention.

    PubMed

    Abbas, Salma; Sastry, Sangeeta

    2016-08-01

    Healthcare-associated infections (HAIs) are an important cause of morbidity and mortality in the USA. They are associated with a substantial increase in health care costs each year. Fortunately, many HAIs are preventable, and their eradication is a national priority. Chlorhexidine (CHG) bathing has been used as an infection prevention measure, either alone or bundled with other interventions, with mostly beneficial results. The recent surge in its use as an agent of choice for skin antisepsis has lead to concerns over emerging resistance among microorganisms. Moreover, compliance with CHG-bathing protocols is not routinely monitored. Policies developed to determine the best infection prevention practice must consider that a "one-size-fits-all" strategy may lead to the selection of CHG-tolerant microorganisms, thereby emphasizing the need for more robust guidelines and additional studies on the role of chlorhexidine bathing for the prevention of HAIs. PMID:27392413

  20. [Pseudomonas folliculitis after spa bath exposure].

    PubMed

    Uldall Pallesen, Kristine Appel; Andersen, Klaus Ejner; Mørtz, Charlotte Gotthard

    2012-06-25

    Pseudomonas aeruginosa is a rare cause of folliculitis. Pseudomonas folliculitis can develop after contact with contaminated water from swimming pools, hot tubs and spa baths. Systemic therapy may be indicated in patients with widespread lesions, systemic symptoms or in immunosuppressed patients. We describe a 23-year-old healthy woman who developed a pustular rash and general malaise after using a spa bath contaminated with Pseudomonas aeruginosa. Bacterial culture from a pustule confirmed Pseudomonas folliculitis and the patient was treated with ciprofloxacin with rapid good effect. PMID:22735119

  1. Mud bath dermatitis due to cinnamon oil.

    PubMed

    García-Abujeta, José Luis; de Larramendi, Carlos Hernando; Berna, José Pomares; Palomino, Elena Muñoz

    2005-04-01

    A case of long-lasting, extensive eczematous and bullous dermatitis affecting exposed areas (arms and legs), beginning within 24 hr after having a mud bath with cinnamon essential oil in a spa, in a 74-year-old woman, is reported. Patch tests with the GEIDC standard battery and the dental battery (including clove essence and eugenol), cinnamon essence and its components were carried out 5 years later. Fragrance mix, cinnamon essence, eugenol, cinnamic alcohol and cinnamic aldehyde yielded a positive result. To our knowledge, this is the first case of cinnamon dermatitis after a mud bath. PMID:15860002

  2. Enhanced photo-fermentative hydrogen production by Rhodobacter capsulatus with pigment content manipulation.

    PubMed

    Ma, Chao; Wang, Xueqing; Guo, Liejin; Wu, Xiaomin; Yang, Honghui

    2012-08-01

    High content of pigment in purple nonsulfur photosynthetic bacteria hinders its photo-hydrogen production rate under intense light irradiation. In order to alleviate the light shielding effect and improve its photo-fermentative hydrogen production performance, pufQ, which is the regulatory gene of bacteriochlorophyll biosynthesis in Rhodobacter capsulatus, was cloned and relocated in the genome under cbb3 promoter by homologous recombination. The UV-vis spectra indicated that the light absorption of the mutant between 300 and 900 nm was reduced. Photo-hydrogen production experiments by the recombinant and wild type strain were carried out in 350 mL photo bioreactors using acetic and butyric acid as substrate. The results showed that the hydrogen production of recombinant with reduced pigment was 27% higher than that of its parental strain, indicating that it is effective on enhancing photo-fermentative hydrogen production by manipulating pigment biosynthesis in purple nonsulfur photosynthetic bacteria. PMID:22717568

  3. In vitro assembly of a prohead-like structure of the Rhodobacter capsulatus gene transfer agent

    SciTech Connect

    Spano, Anthony J. . E-mail: ajs6z@virginia.edu; Chen, Frank S.; Goodman, Benjamin E.; Sabat, Agnes E.; Simon, Martha N.; Wall, Joseph S.; Correia, John J.; McIvor, Wilson; Newcomb, William W.; Brown, Jay C.; Schnur, Joel M.; Lebedev, Nikolai

    2007-07-20

    The gene transfer agent (GTA) is a phage-like particle capable of exchanging double-stranded DNA fragments between cells of the photosynthetic bacterium Rhodobacter capsulatus. Here we show that the major capsid protein of GTA, expressed in E. coli, can be assembled into prohead-like structures in the presence of calcium ions in vitro. Transmission electron microscopy (TEM) of uranyl acetate staining material and thin sections of glutaraldehyde-fixed material demonstrates that these associates have spherical structures with diameters in the range of 27-35 nm. The analysis of scanning TEM images revealed particles of mass {approx} 4.3 MDa, representing 101 {+-} 11 copies of the monomeric subunit. The establishment of this simple and rapid method to form prohead-like particles permits the GTA system to be used for genome manipulation within the photosynthetic bacterium, for specific targeted drug delivery, and for the construction of biologically based distributed autonomous sensors for environmental monitoring.

  4. In vitro assembly of a prohead-like structure of the Rhodobacter capsulatus gene transfer agent.

    PubMed

    Spano, Anthony J; Chen, Frank S; Goodman, Benjamin E; Sabat, Agnes E; Simon, Martha N; Wall, Joseph S; Correia, John J; McIvor, Wilson; Newcomb, William W; Brown, Jay C; Schnur, Joel M; Lebedev, Nikolai

    2007-07-20

    The gene transfer agent (GTA) is a phage-like particle capable of exchanging double-stranded DNA fragments between cells of the photosynthetic bacterium Rhodobacter capsulatus. Here we show that the major capsid protein of GTA, expressed in E. coli, can be assembled into prohead-like structures in the presence of calcium ions in vitro. Transmission electron microscopy (TEM) of uranyl acetate staining material and thin sections of glutaraldehyde-fixed material demonstrates that these associates have spherical structures with diameters in the range of 27-35 nm. The analysis of scanning TEM images revealed particles of mass approximately 4.3 MDa, representing 101+/-11 copies of the monomeric subunit. The establishment of this simple and rapid method to form prohead-like particles permits the GTA system to be used for genome manipulation within the photosynthetic bacterium, for specific targeted drug delivery, and for the construction of biologically based distributed autonomous sensors for environmental monitoring. PMID:17408713

  5. Cloning, characterization, and regulation of nifF from Rhodobacter capsulatus.

    PubMed

    Gennaro, G; Hübner, P; Sandmeier, U; Yakunin, A F; Hallenbeck, P C

    1996-07-01

    The Rhodobacter capsulatus nifF gene and upstream sequence were cloned by using a probe based on the N-terminal sequence of NifF. nifF was found to not be contained in the previously described nif regions I, II, and III. Comparison of the deduced amino acid sequence showed that it is highly similar to NifF from Azotobacter vinelandii and NifF from Klebsiella pneumoniae. Analysis of translational fusions demonstrated that the regulation of transcription was the same as previously reported at the protein level. Insertional mutagen esis showed that NifF contributes significantly to nitrogenase activity under normal nitrogen-fixing conditions and that it is absolutely required for nitrogen fixation under iron limitation. PMID:8682802

  6. Transcriptional and Posttranscriptional Events Control Copper-Responsive Expression of a Rhodobacter capsulatus Multicopper Oxidase

    PubMed Central

    Rademacher, Corinna; Moser, Roman; Lackmann, Jan-Wilm; Klinkert, Birgit; Narberhaus, Franz

    2012-01-01

    The copper-regulated Rhodobacter capsulatus cutO (multicopper oxidase) gene confers copper tolerance and is carried in the tricistronic orf635-cutO-cutR operon. Transcription of cutO strictly depends on the promoter upstream of orf635, as demonstrated by lacZ reporter fusions to nested promoter fragments. Remarkably, orf635 expression was not affected by copper availability, whereas cutO and cutR were expressed only in the presence of copper. Differential regulation was abolished by site-directed mutations within the orf635-cutO intergenic region, suggesting that this region encodes a copper-responsive mRNA element. Bioinformatic predictions and RNA structure probing experiments revealed an intergenic stem-loop structure as the candidate mRNA element. This is the first posttranscriptional copper response mechanism reported in bacteria. PMID:22287514

  7. Replacement of sugars to hydrogen production by Rhodobacter capsulatus using dark fermentation effluent as substrate.

    PubMed

    Silva, Felipe Thales Moreira; Moreira, Luiza Rojas; de Souza Ferreira, Juliana; Batista, Fabiana Regina Xavier; Cardoso, Vicelma Luiz

    2016-01-01

    Hydrogen is a promising alternative for the increased global energy demand since it has high energy density and is a clean fuel. The aim of this work was to evaluate the photo-fermentation by Rhodobacter capsulatus, using the dark fermentation effluent as substrate. Different systems were tested by changing the type of sugar in the dark fermentation, investigating the influence of supplementing DFE with sugar and adding alternate and periodically lactose and glucose throughout the process. The supplementation of the DFE with sugar resulted in higher H2 productivity and the replacement of the sugars repeatedly during the photo-fermentation process was important to maintain the cell culture active. By controlling the residual amount of sugar, bacteria inhibition was avoided; lactic acid, that was toxic to the biomass, was consumed and the metabolic route of butyric acid production was predominant. Under optimum conditions, the H2 productivity reached 208.40mmolH2/Ld in 52h. PMID:26476167

  8. [Surface layers of methanotrophic bacteria].

    PubMed

    Khmelenina, V N; Suzina, N E; Trotsenko, Iu A

    2013-01-01

    Structural and functional characteristics of the regular glycoprotein layers in prokaryotes are analyzed with a special emphasis on aerobic methanotrophic bacteria. S-layers are present at the surfaces of Methylococcus, Methylothermus, and Methylomicrobium cells. Different Methylomicrobium species either synthesize S-layers with planar (p2, p4) symmetry or form cup-shaped or conicalstructures with hexagonal (p6) symmetry. A unique, copper-binding polypeptide 'CorA'/MopE (27/45 kDa), which is coexpressed with the diheme periplasmic cytochrome c peroxidase 'CorB'/Mca (80 kDa) was found in Methylomicrobium album BG8, Methylomicrobium alcaliphilum 20Z, and Methylococcus capsulatus Bath. This tandem of the surface proteins is functionally analogous to a new siderophore, methanobactin. Importantly, no 'CorA'/MopE homologue was found in methanotrophs not forming S-layers. The role of surface proteins in copper metabolism and initial methane oxidation is discussed. PMID:25509389

  9. Molecular genetic and molecular evolutionary studies on the bacteriochlorophyll synthesis genes of Rhodobacter capsulatus

    SciTech Connect

    Burke-Agueero, D.H.

    1992-08-01

    Rhodobacter capsulatus, purple bacterium capable of either aerobic or photosynthetic growth, has proven to be very useful in genetic studies of photosynthesis. Forty-four genes clustered together within a 46 kilobase region are required to establish photosynthetic ability in R. capsulatus. Approximately twenty of these genes are involved in bacteriochlorophyll synthesis of which eight ``bch`` genes are the subject of this thesis. Six of these genes were found to code for the two ring reductases. The first converts protochlorophyllide (PChlide) into a chlorin, the immediate precursor to chlorophyll a, and then into a bacteriochlorin. Each reductase is shown to be made up of three subunits. PChlide reductase is coded by the genes bchN, bchB, and bchL. Proteins with amino acid sequences markedly similar to those of bchN and bchL have been shown in other organisms to be required for chlorophyll synthesis; hence, their designation as chlN and chlB. A third chloroplast-encoded gene of heretofore unknown function shares amino acid identities with bchB and is probably the third subunit of the plant PChlide reductase. The bchA locus, which encodes the chlorin reductase, is found to be made up of three separate, translationally coupled genes, referred to as bchX, bchY, and bchZ. Amino acid similarities between bchX, bchL, and the nitrogenase reductase protein nifH suggest that all three classes of proteins share certain three-dimensional structural features, including elements that are central to the enzymatic mechanism of nifH. PChlide reductase and chlorin reductase are clearly derived from a common ancestor. Several lines of analysis suggests the ancestor of both enzyme systems reduced PChlide twice to produce bacteriochlorophyll supporting the concept bacteriochlorophyll as the ancestral reaction center pigment.

  10. Molecular genetic and molecular evolutionary studies on the bacteriochlorophyll synthesis genes of Rhodobacter capsulatus

    SciTech Connect

    Burke-Agueero, D.H.

    1992-08-01

    Rhodobacter capsulatus, purple bacterium capable of either aerobic or photosynthetic growth, has proven to be very useful in genetic studies of photosynthesis. Forty-four genes clustered together within a 46 kilobase region are required to establish photosynthetic ability in R. capsulatus. Approximately twenty of these genes are involved in bacteriochlorophyll synthesis of which eight bch'' genes are the subject of this thesis. Six of these genes were found to code for the two ring reductases. The first converts protochlorophyllide (PChlide) into a chlorin, the immediate precursor to chlorophyll a, and then into a bacteriochlorin. Each reductase is shown to be made up of three subunits. PChlide reductase is coded by the genes bchN, bchB, and bchL. Proteins with amino acid sequences markedly similar to those of bchN and bchL have been shown in other organisms to be required for chlorophyll synthesis; hence, their designation as chlN and chlB. A third chloroplast-encoded gene of heretofore unknown function shares amino acid identities with bchB and is probably the third subunit of the plant PChlide reductase. The bchA locus, which encodes the chlorin reductase, is found to be made up of three separate, translationally coupled genes, referred to as bchX, bchY, and bchZ. Amino acid similarities between bchX, bchL, and the nitrogenase reductase protein nifH suggest that all three classes of proteins share certain three-dimensional structural features, including elements that are central to the enzymatic mechanism of nifH. PChlide reductase and chlorin reductase are clearly derived from a common ancestor. Several lines of analysis suggests the ancestor of both enzyme systems reduced PChlide twice to produce bacteriochlorophyll supporting the concept bacteriochlorophyll as the ancestral reaction center pigment.

  11. The Reductive Half-reaction of Xanthine Dehydrogenase from Rhodobacter capsulatus

    PubMed Central

    Hall, James; Reschke, Stefan; Cao, Hongnan; Leimkühler, Silke; Hille, Russ

    2014-01-01

    The kinetic properties of an E232Q variant of the xanthine dehydrogenase from Rhodobacter capsulatus have been examined to ascertain whether Glu232 in wild-type enzyme is protonated or unprotonated in the course of catalysis at neutral pH. We find that kred, the limiting rate constant for reduction at high [xanthine], is significantly compromised in the variant, a result that is inconsistent with Glu232 being neutral in the active site of the wild-type enzyme. A comparison of the pH dependence of both kred and kred/Kd from reductive half-reaction experiments between wild-type and enzyme and the E232Q variant suggests that the ionized Glu232 of wild-type enzyme plays an important role in catalysis by discriminating against the monoanionic form of substrate, effectively increasing the pKa of substrate by two pH units and ensuring that at physiological pH the neutral form of substrate predominates in the Michaelis complex. A kinetic isotope study of the wild-type R. capsulatus enzyme indicates that, as previously determined for the bovine and chicken enzymes, product release is principally rate-limiting in catalysis. The disparity in rate constants for the chemical step of the reaction and product release, however, is not as great in the bacterial enzyme as compared with the vertebrate forms. The results indicate that the bacterial and bovine enzymes catalyze the chemical step of the reaction to the same degree and that the faster turnover observed with the bacterial enzyme is due to a faster rate constant for product release than is seen with the vertebrate enzyme. PMID:25258317

  12. [Is Turkish bath water potable?: The baths of Sidi-Bel-Abbes].

    PubMed

    Benouis, K; Benabderrahmane, M; Harrache-Chettouh, Djamila; Benabdeli, K

    2008-01-01

    In Algeria, large numbers of people regularly go to Turkish baths or "Hammams". The cold tap water of the baths in the town of Sidi-Bel-Abbes (Algeria) comes either from wells or from a mixture of potable waterworks water and well water. Its principal use is for personal hygiene (washing). However, the steam heat generates thirst that can cause users to drink cold water during the steam bath. In addition, the wells feeding the baths are often poorly protected and especially badly treated. To ascertain whether their water quality, particularly bacteriological, meets the requirements for drinking water, we studied the characteristics of water from ten Turkish baths in Sidi-Bel-Abbes. Bacteriological analyses of cold water showed signs of contamination of fecal origin in 50% of the samples analysed. Moreover two water points from two of the baths appeared to have permanent fecal contamination. The physicochemical analysis showed that the water was very high in calcium (up to 550 mg/L) and magnesium (up to 299 mg/L). The maximum nitrate level observed was 68 mg/L. This study thus showed the existence of a health risk due to deterioration in the quality of the bath water and demonstrated the need for protection of the wells, frequent purification, and regular microbiological testing. PMID:19188127

  13. Metal/protein ratio determination in the Rhodobacter capsulatus cytoplasmic pyrophosphatase enzyme by particle induced X-ray emission.

    PubMed

    Solís, C; Celis, H; Romero, I; Isaac-Olivé, K; Andrade, E; Eslava-Rosas, F; Monroy-Rosales, J H; Cuevas-Moreno, R

    2011-02-01

    Inorganic pyrophosphatases are divided in two families, which differ both in structure and mechanism. All of them incorporate in its structure divalent metal cations. In 2003, it was reported for the first time that Rhodobacter capsulatus cytoplasmic pyrophosphatase belongs to family II. It is expected then, that this enzyme contains metal elements in its structure; however, this characterization has not been carried out yet. A fine application of accelerators is the use of proton beams to induce X-ray emission (PIXE) for analyzing the composition of biological macromolecules. The purpose of this work is to complement R. capsulatus cytoplasmic pyrophosphatase characterization by determining the presence of metal elements in its structure. Three different strategies were used: PAGE-PIXE, PAGE-Digestion-PIXE, and Dialysis-PIXE and when metals were found the metal/enzyme ratio was calculated. Only cobalt was found to be associated to the enzyme chemical structure in a ratio 3 Co/enzyme. PMID:21167875

  14. Decoherence Dynamics of a Superconducting Charge Qubit Coupled to a Boson Bath and a Spin Bath

    NASA Astrophysics Data System (ADS)

    Yang, Qin-Ying; Liang, Bao-Long; Wang, Ji-Suo

    2014-01-01

    By virtue of a superconducting charge qubit, we derive the off-diagonal matrix operator and investigate the decoherence of the system in different regimes coupled to, respectively, the boson bath and the spin bath. It is found that the two different baths make a bit of difference on the decay of the system at low but finite temperature and the decoherence of the system is most closely linked with the regime as well as the coupling strength. Therefore, by optimizing some reasonable parameters, we can suppress appropriately the decoherence of a given quantum system.

  15. Inactivation, sequence, and lacZ fusion analysis of a regulatory locus required for repression of nitrogen fixation genes in Rhodobacter capsulatus.

    PubMed Central

    Kranz, R G; Pace, V M; Caldicott, I M

    1990-01-01

    Transcription of the genes that code for proteins involved in nitrogen fixation in free-living diazotrophs is typically repressed by high internal oxygen concentrations or exogenous fixed nitrogen. The DNA sequence of a regulatory locus required for repression of Rhodobacter capsulatus nitrogen fixation genes was determined. It was shown that this locus, defined by Tn5 insertions and by ethyl methanesulfonate-derived mutations, is homologous to the glnB gene of other organisms. The R. capsulatus glnB gene was upstream of glnA, the gene for glutamine synthetase, in a glnBA operon. beta-Galactosidase expression from an R. capsulatus glnBA-lacZ translational fusion was increased twofold in cells induced by nitrogen limitation relative to that in cells under nitrogen-sufficient conditions. R. capsulatus nifR1, a gene that was previously shown to be homologous to ntrC and that is required for transcription of nitrogen fixation genes, was responsible for approximately 50% of the transcriptional activation of this glnBA fusion in cells induced under nitrogen-limiting conditions. R. capsulatus GLNB, NIFR1, and NIFR2 (a protein homologous to NTRB) were proposed to transduce the nitrogen status in the cell into repression or activation of other R. capsulatus nif genes. Repression of nif genes in response to oxygen was still present in R. capsulatus glnB mutants and must have occurred at a different level of control in the regulatory circuit. Images FIG. 4 FIG. 5 PMID:2152916

  16. Bubble bath burns: an unusual case

    PubMed Central

    Nizamoglu, Metin; Tan, Alethea; El-Muttardi, Naguib

    2016-01-01

    Abstract We present an unusual case of flash burn injury in an adolescent following accidental combination of foaming bath bubbles and tea light candle flame. There has not been any reported similar case described before. This serves as a learning point for public prevention and clinicians managing burn injuries. PMID:27583271

  17. Bubble bath burns: an unusual case.

    PubMed

    Nizamoglu, Metin; Tan, Alethea; El-Muttardi, Naguib

    2016-01-01

    We present an unusual case of flash burn injury in an adolescent following accidental combination of foaming bath bubbles and tea light candle flame. There has not been any reported similar case described before. This serves as a learning point for public prevention and clinicians managing burn injuries. PMID:27583271

  18. String melting in a photon bath

    SciTech Connect

    Karouby, Johanna

    2013-10-01

    We compute the decay rate of a metastable cosmic string in contact with a thermal bath by finding the instanton solution. The new feature is that this decay rate is found in the context of non thermal scalar fields in contact with a thermal bath of photons. In general, to make topologically unstable strings stable, one can couple them to such a bath. The resulting plasma effect creates metastable configurations which can decay from the false vacuum to the true vacuum. In our specific set-up, the instanton computation is realized for the case of two out-of-equilibrium complex scalar fields: one is charged and coupled to the photon field, and the other is neutral. New effects coming from the thermal bath of photons make the radius of the nucleated bubble and most of the relevant physical quantities temperature-dependent. However, the temperature appears in a different way than in the purely thermal case, where all scalar fields are in thermal equilibrium. As a result of the tunneling, the core of the initial string melts while bubbles of true vacuum expand at the speed of light.

  19. Chromosomal structure of Rhodobacter capsulatus strain SB1003: Cosmid encyclopedia and high-resolution physical and genetic map

    SciTech Connect

    Fonstein, M.; Haselkorn, R. )

    1993-03-15

    A combination of cosmid genome walking and pulsed-field gel electrophoresis was used to construct a high-resolution physical and genetic map of the 3.8-megabase (Mb) genome of Rhodobacter capsulatus SB1003. The mapping was done by hybridization of pulsed-field gel blots and by grouping and further mapping of the cosmids and bacteriophages from genomic libraries. Cosmid clones formed two uninterrupted and ordered groups, one corresponding to the chromosome of R. capsulatus, the other to its 134-kb plasmid. Cos site end-labeling and partial EcoRV digestion of cosmids were used to construct a high-resolution EvoRV map of the genome. Overlapping of the mosmids was confirmed by the resemblance of the cosmid restriction maps and by direct end-to-end hybridization with SP6- and T7-specific transcripts. Twenty-three previously cloned genes and eight groups of repeated sequences, revealed in this work, were located in the ordered gene library and mapped with an accuracy of 1-10 kb. Blots of a minimal set of 192 cosmids, covering the chromosome and the plasmid with the known map position of each cosmid, give to R. capsulatus the same advantages that the Kohara phage panel gives to E. coli. 15 refs., 4 figs.

  20. Psychoactive "bath salts": not so soothing.

    PubMed

    Baumann, Michael H; Partilla, John S; Lehner, Kurt R

    2013-01-01

    Recently there has been a dramatic rise in the abuse of so-called "bath salts" products that are purchased as legal alternatives to illicit drugs like cocaine and 3,4-methylenedioxymethamphetamine (MDMA). Baths salts contain one or more synthetic derivatives of the naturally-occurring stimulant cathinone. Low doses of bath salts produce euphoria and increase alertness, but high doses or chronic use can cause serious adverse effects such as hallucinations, delirium, hyperthermia and tachycardia. Owing to the risks posed by bath salts, the governments of many countries have made certain cathinones illegal, namely: 4-methylmethcathinone (mephedrone), 3,4-methylenedioxymethcathinone (methylone) and 3,4-methylenedioxypyrovalerone (MDPV). Similar to other psychomotor stimulants, synthetic cathinones target plasma membrane transporters for dopamine (i.e., DAT), norepinephrine (i.e., NET) and serotonin (i.e, SERT). Mephedrone and methylone act as non-selective transporter substrates, thereby stimulating non-exocytotic release of dopamine, norepinephrine and serotonin. By contrast, MDPV acts as a potent blocker at DAT and NET, with little effect at SERT. Administration of mephedrone or methylone to rats increases extracellular concentrations of dopamine and serotonin in the brain, analogous to the effects of MDMA. Not surprisingly, synthetic cathinones elicit locomotor activation in rodents. Stimulation of dopamine transmission by synthetic cathinones predicts a high potential for addiction and may underlie clinical adverse effects. As popular synthetic cathinones are rendered illegal, new replacement cathinones are appearing in the marketplace. More research on the pharmacology and toxicology of abused cathinones is needed to inform public health policy and develop strategies for treating medical consequence of bath salts abuse. PMID:23178799

  1. General view, Belair Bath and Tennis Club, Belair at Bowie, ...

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

    General view, Belair Bath and Tennis Club, Belair at Bowie, Maryland, looking west. - Belair Bath and Tennis Club, Southwest corner of Belair Drive and Tulip Grove Drive, Bowie, Prince George's County, MD

  2. 20 CFR 654.412 - Bathing, laundry, and handwashing.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... Bathing, laundry, and handwashing. (a) Bathing and handwashing facilities, supplied with hot and cold... facilities, supplied with hot and cold water under pressure, shall be provided for the use of all...

  3. Recombinant Rhodobacter capsulatus xanthine dehydrogenase, a useful model system for the characterization of protein variants leading to xanthinuria I in humans.

    PubMed

    Leimkuhler, Silke; Hodson, Rachael; George, Graham N; Rajagopalan, K V

    2003-06-01

    Rhodobacter capsulatus xanthine dehydrogenase (XDH) forms an (alphabeta)2 heterotetramer and is highly homologous to homodimeric eukaryotic XDHs. The crystal structures of bovine XDH and R. capsulatus XDH showed that the two proteins have highly similar folds. We have developed an efficient system for the recombinant expression of R. capsulatus XDH in Escherichia coli. The recombinant protein shows spectral features and a range of substrate specificities similar to bovine milk xanthine oxidase. However, R. capsulatus XDH is at least 5 times more active than bovine XDH and, unlike mammalian XDH, does not undergo the conversion to the oxidase form. EPR spectra were obtained for the FeS centers of the enzyme showing an axial signal for FeSI, which is different from that reported for xanthine oxidase. X-ray absorption spectroscopy at the iron and molybdenum K-edge and the tungsten LIII-edge have been used to probe the different metal coordinations of variant forms of the enzyme. Based on a mutation identified in a patient suffering from xanthinuria I, the corresponding arginine 135 was substituted to a cysteine in R. capsulatus XDH, and the protein variant was purified and characterized. Two different forms of XDH-R135C were purified, an active (alphabeta)2 heterotetrameric form and an inactive (alphabeta) heterodimeric form. The active form contains a full complement of redox centers, whereas in the inactive form the FeSI center is likely to be missing. PMID:12670960

  4. Processing A Printed Wiring Board By Single Bath Electrodeposition

    DOEpatents

    Meltzer, Michael P.; Steffani, Christopher P.; Gonfiotti, Ray A.

    2003-04-15

    A method of processing a printed wiring board by single bath electrodeposition. Initial processing steps are implemented on the printed wiring board. Copper is plated on the printed wiring board from a bath containing nickel and copper. Nickel is plated on the printed wiring board from the bath containing nickel and copper and final processing steps are implemented on the printed wiring board.

  5. Autogenous grinding for bath scraps recycling

    SciTech Connect

    Pinoncely, A.; Podda, P.

    1996-10-01

    In the early 80`s, FCB designed an original process for the recycling of bath scraps in Aluminum smelters, using a single stage fully air-swept autogenous mill. Since then, the 9 industrial references confirmed and even exceeded the expectation in terms of dust-free and easy to run operation, high recovery ratio of bath among the metallic scraps, and low maintenance cost. Problems encountered on conventional processes belong to the old days, and new projects tend to give an increasing importance to classification and storage of crushed products, autogenous grinding being already recognized as the most suitable, simple and reliable process route. The present paper describes this original process and draws up the overall performances of ten years of experience.

  6. Astronaut Jack Lousma taking hot bath

    NASA Technical Reports Server (NTRS)

    1973-01-01

    A closeup view of Astronaut Jack R. Lousma, Skylab 3 pilot, taking a hot bath in the crew quarters of the Orbital Workshop (OWS) of the Skylab space station cluster in Earth orbit. In deploying the shower facility, the shower curtain is pulled up from the floor and attached to the ceiling. The water comes through a push-button shower head attached to a flexible hose. Water is drawn off by a vacuum system.

  7. Leidenfrost drops on liquid baths: theory

    NASA Astrophysics Data System (ADS)

    Sobac, Benjamin; Rednikov, Alexei; Maquet, Laurent; Darbois-Texier, Baptiste; Duchesne, Alexis; Brandenbourger, Martin; Dorbolo, Stéphane; Colinet, Pierre

    2015-11-01

    It is well known that a liquid drop released over a very hot surface generally does not contact the surface nor boils but rather levitates over a thin vapor film generated by its own evaporation (Leidenfrost effect). In particular, the case of a hot (and flat) solid substrate has been extensively studied in recent years. In contrast, we here focus on Leidenfrost drops over a superheated liquid bath, addressing the problem theoretically and comparing our predictions with experimental results, detailed in a separate talk. We predict the geometry of the drop and of the liquid bath, based on the hydrostatic Young-Laplace and lubrication equations. A good agreement is observed with the available experimental data concerning the deformation of the liquid bath. The modeling also yields a rather complete insight into the shape of the drop. As in the case of a solid substrate, the vapor layer generally appears to be composed of a vapor pocket surrounded by a circular neck. The influences of the superheat and of the drop size are parametrically investigated. A number of scaling laws are established. Unlike the case of a solid substrate, no chimney instability was found in the range of drop size studied.

  8. Molten salt bath circulation design for an electrolytic cell

    DOEpatents

    Dawless, Robert K.; LaCamera, Alfred F.; Troup, R. Lee; Ray, Siba P.; Hosler, Robert B.

    1999-01-01

    An electrolytic cell for reduction of a metal oxide to a metal and oxygen has an inert anode and an upwardly angled roof covering the inert mode. The angled roof diverts oxygen bubbles into an upcomer channel, thereby agitating a molten salt bath in the upcomer channel and improving dissolution of a metal oxide in the molten salt bath. The molten salt bath has a lower velocity adjacent the inert anode in order to minimize corrosion by substances in the bath. A particularly preferred cell produces aluminum by electrolysis of alumina in a molten salt bath containing aluminum fluoride and sodium fluoride.

  9. Protecting coherence by reservoir engineering: intense bath disturbance

    NASA Astrophysics Data System (ADS)

    Zhou, Zixian; Lü, Zhiguo; Zheng, Hang

    2016-08-01

    We put forward a scheme based on reservoir engineering to protect quantum coherence from leaking to bath, in which we intensely disturb the Lorentzian bath by N harmonic oscillators. We show that the intense disturbance changes the spectrum of the bath and reduces the qubit-bath interaction. Furthermore, we give the exact time evolution with the Lorentzian spectrum by a master equation and calculate the concurrence and survival probability of the qubits to demonstrate the effect of the intense bath disturbance on the protection of coherence. Meanwhile, we reveal the dynamic effects of counter-rotating interaction on the qubits as compared to the results of the rotating-wave approximation.

  10. Molten salt bath circulation design for an electrolytic cell

    DOEpatents

    Dawless, R.K.; LaCamera, A.F.; Troup, R.L.; Ray, S.P.; Hosler, R.B.

    1999-08-17

    An electrolytic cell for reduction of a metal oxide to a metal and oxygen has an inert anode and an upwardly angled roof covering the inert mode. The angled roof diverts oxygen bubbles into an upcomer channel, thereby agitating a molten salt bath in the upcomer channel and improving dissolution of a metal oxide in the molten salt bath. The molten salt bath has a lower velocity adjacent the inert anode in order to minimize corrosion by substances in the bath. A particularly preferred cell produces aluminum by electrolysis of alumina in a molten salt bath containing aluminum fluoride and sodium fluoride. 4 figs.

  11. Protecting coherence by reservoir engineering: intense bath disturbance

    NASA Astrophysics Data System (ADS)

    Zhou, Zixian; Lü, Zhiguo; Zheng, Hang

    2016-05-01

    We put forward a scheme based on reservoir engineering to protect quantum coherence from leaking to bath, in which we intensely disturb the Lorentzian bath by N harmonic oscillators. We show that the intense disturbance changes the spectrum of the bath and reduces the qubit-bath interaction. Furthermore, we give the exact time evolution with the Lorentzian spectrum by a master equation and calculate the concurrence and survival probability of the qubits to demonstrate the effect of the intense bath disturbance on the protection of coherence. Meanwhile, we reveal the dynamic effects of counter-rotating interaction on the qubits as compared to the results of the rotating-wave approximation.

  12. Model of the molecular basis for hydroxylamine oxidation and nitrous oxide production in methanotrophic bacteria.

    PubMed

    Campbell, Mark A; Nyerges, Györgyi; Kozlowski, Jessica A; Poret-Peterson, Amisha T; Stein, Lisa Y; Klotz, Martin G

    2011-09-01

    Many methane-oxidizing bacteria (MOB) have been shown to aerobically oxidize ammonia and hydroxylamine (NH(2)OH) to produce nitrite and nitrous oxide (N(2)O). Genome sequences of alphaproteobacterial, gammaproteobacterial, and verrucomicrobial methanotrophs revealed the presence of haoAB, cytL, cytS, nirS or nirK, and norCB genes that may be responsible for N(2)O production, and additional haoAB genes were sequenced from two strains of Methylomicrobium album. The haoAB genes of M. album ATCC 33003 were inducible by ammonia and NH(2)OH, similar to haoAB induction by ammonia in Methylococcus capsulatus Bath. Increased expression of genes encoding nitric oxide reductase (cNOR; norCB) was measured upon exposure of M. capsulatus Bath to NaNO(2) and NO-releasing sodium nitroprusside. Only incubations of M. capsulatus Bath with methane, ammonia, and nitrite produced N(2)O. The data suggest a possible pathway of nitrite reduction to NO by reversely operating NH(2)OH oxidoreductase and NO reduction to N(2)O by cNOR independently or in conjunction with ammonia-induced enzymes (i.e. HAO or cytochrome c'-β). Results of this study show that MOB likely have diverse mechanisms for nitrogen oxide metabolism and detoxification of NH(2)OH that involve conventional and unconventional enzymes. PMID:21682764

  13. Heat-tolerant methanotropic bacteria from the hot water effluent of a natural gas field

    SciTech Connect

    Bodrossy, L.; Kovacs, K.L.; Murrell, C.

    1995-10-01

    Methanotrophic bacteria were isolated from a natural environment potentially favorable to heat-tolerant methanotrophs. An improved colony plate assay was developed and used to identify putative methanotrophic colonies with high confidence. Fourteen new isolates were purified and partially characterized. These new isolates exhibit a DNA sequence homology of up to 97% with the conserved regions in the mmoX and mmoC genes of the soluble methane monooxygenase (MMO)-coding gene cluster of Methylococcus capsulatus Bath. The copper regulations of soluble MMO expression in the same isolates, however, differs from that of M. capsulatus Bath, as the new isolates can tolerate up to 0.8 {mu}M copper without loss of MMO activity while a drastic reduction of MMO activity occurs already at 0.1 {mu}M copper in M. capsulatus Bath. The isolates can be cultivated and utilized at elevated temperatures, and their copper- and heat-tolerant MMO activity makes these bacterial ideal candidates for future biotechnological use. 35 refs., 6 figs.

  14. Role of initial system-bath correlation on coherence trapping

    PubMed Central

    Zhang, Ying-Jie; Han, Wei; Xia, Yun-Jie; Yu, Yan-Mei; Fan, Heng

    2015-01-01

    We study the coherence trapping of a qubit correlated initially with a non-Markovian bath in a pure dephasing channel. By considering the initial qubit-bath correlation and the bath spectral density, we find that the initial qubit-bath correlation can lead to a more efficient coherence trapping than that of the initially separable qubit-bath state. The stationary coherence in the long time limit can be maximized by optimizing the parameters of the initially correlated qubit-bath state and the bath spectral density. In addition, the effects of this initial correlation on the maximal evolution speed for the qubit trapped to its stationary coherence state are also explored. PMID:26303160

  15. Triterpene hydrocarbon production engineered into a metabolically versatile host--Rhodobacter capsulatus.

    PubMed

    Khan, Nymul E; Nybo, S Eric; Chappell, Joe; Curtis, Wayne R

    2015-08-01

    Triterpene hydrocarbon biosynthesis of the ancient algae Botryococcus braunii was installed into Rhodobacter capsulatus to explore the production of C30 hydrocarbon in a host capable of diverse growth habits-utilizing carbohydrate, sunlight or hydrogen (with CO2 fixation) as alternative energy feedstocks. Engineering an enhanced MEP pathway was also used to augment triterpene accumulation. Despite dramatically different sources of carbon and reducing power, nearly the same level of botryococcene or squalene (∼5 mg oil/g-dry-weight [gDW]) was achieved in small-scale aerobic heterotrophic, anaerobic photoheterotrophic, and aerobic chemoautotrophic growth conditions. A glucose fed-batch bioreactor reached 40 mg botryococcene/L (∼12 mg/gDW), while autotrophic bioreactor performance with CO2 , H2 , and O2 reached 110 mg/L (16.7 mg/gDW) during batch and 60 mg/L (23 mg/gDW) during continuous operation at a dilution rate corresponding to about 10% of μ(max). Batch and continuous autotrophic specific productivity was found to reach 0.5 and 0.32 mg triterpene/g DW/h, comparable to prior reports for terpene production driven by heterotrophic growth conditions. This demonstrates the feasibility of alternative feedstocks and trophic modes to provide comparable routes to biochemicals that do not rely on sugar. PMID:25728701

  16. Two distinct ferredoxins from Rhodobacter capsulatus: complete amino acid sequences and molecular evolution.

    PubMed

    Saeki, K; Suetsugu, Y; Yao, Y; Horio, T; Marrs, B L; Matsubara, H

    1990-09-01

    Two distinct ferredoxins were purified from Rhodobacter capsulatus SB1003. Their complete amino acid sequences were determined by a combination of protease digestion, BrCN cleavage and Edman degradation. Ferredoxins I and II were composed of 64 and 111 amino acids, respectively, with molecular weights of 6,728 and 12,549 excluding iron and sulfur atoms. Both contained two Cys clusters in their amino acid sequences. The first cluster of ferredoxin I and the second cluster of ferredoxin II had a sequence, CxxCxxCxxxCP, in common with the ferredoxins found in Clostridia. The second cluster of ferredoxin I had a sequence, CxxCxxxxxxxxCxxxCM, with extra amino acids between the second and third Cys, which has been reported for other photosynthetic bacterial ferredoxins and putative ferredoxins (nif-gene products) from nitrogen-fixing bacteria, and with a unique occurrence of Met. The first cluster of ferredoxin II had a CxxCxxxxCxxxCP sequence, with two additional amino acids between the second and third Cys, a characteristics feature of Azotobacter-[3Fe-4S] [4Fe-4S]-ferredoxin. Ferredoxin II was also similar to Azotobacter-type ferredoxins with an extended carboxyl (C-) terminal sequence compared to the common Clostridium-type. The evolutionary relationship of the two together with a putative one recently found to be encoded in nifENXQ region in this bacterium [Moreno-Vivian et al. (1989) J. Bacteriol. 171, 2591-2598] is discussed. PMID:2277040

  17. Effects of cadmium stress on growth, morphology, and protein expression in Rhodobacter capsulatus B10.

    PubMed

    Mohamed Fahmy Gad El-Rab, Sanaa; Abdel-Fattah Shoreit, Ahmed; Fukumori, Yoshihiro

    2006-10-01

    The effects of cadmium stress on growth, morphology, and protein expression were investigated in Rhodobacter capsulatus B10 using two-dimensional polyacrylamide gel electrophoresis and a scanning electron microscope with an energy dispersive X-ray spectrometer. The bacterium grew in the presence of 150 microM CdCl2 and highly induced heat-shock proteins (GroEL and Dnak), S-adenosylmethionine synthetase, ribosomal protein S1, aspartate aminotransferase, and phosphoglycerate kinase. Interestingly, the ribosomal protein S1 was proportionally expressed as the amount of cadmium in the medium, suggesting that S1 may be required for the repair of cadmium-mediated cellular damage. On the other hand, we identified five cadmium-binding proteins: 2-methylcitrate dehydratase, phosphate periplasmic binding protein, inosine-5'-monophosphate dehydrogenase/guanosine-5'-monophosphate reductase, inositol monophosphatase, and lytic murein transglycosylase. The cadmium-treated cells had a filamentous structure and contained less phosphorous than the untreated cells. We propose that these characteristics of the cadmium-treated cells may be due to the inactivation of the phosphate periplasmic binding protein and lytic murein transglycosylase by cadmium. PMID:17031048

  18. A molecular genetic analysis of carotenoid biosynthesis and the effects of carotenoid mutations on other photosynthetic genes in Rhodobacter capsulatus

    SciTech Connect

    Armstrong, G.A.

    1989-04-01

    The nine known R. capsulatus carotenoid genes are contained within the 46 kilobase (kb) photosynthesis gene cluster. An 11 kb subcluster containing eight of these genes has been cloned and its nucleotide sequence determined. A new gene, crtK, has been located in the middle of the subcluster. The carotenoid gene cluster contains sequences homologous to Escherichia coli ..omega../sup 70/ promoters, rho-independent transcription terminators, and prokaryotic transcriptional factor binding sites. The phenotypes and genotypes of ten transposon Tn5.7 insertion mutations within the carotenoid gene cluster have been analyzed, by characterization of the carotenoids accumulated and high resolution mapping of the Tn5.7 insertions. The enzymatic blockages in previously uncharacterized early carotenoid mutants have been determined using a new in vitro synthesis system, suggesting specific roles for the CrtB and CrtE gene products. The expression of six of the eight carotenoid genes in the cluster is induced upon the shift from dark chemoheterotrophic to anaerobic photosynthetic growth. The magnitude of the induction is equivalent to that of genes encoding structural photosynthesis polypeptides, although the carotenoid genes are induced earlier after the growth shift. Different means of regulating photosynthesis genes in R. capsulatus are discussed, and a rationale for the temporal pattern of expression of the carotenoid genes during photosynthetic adaptation is presented. Comparison of the deduced amino acid sequences of the two dehydrogenases of the R. capsulatus carotenoid biosynthesis pathway reveals two regions of strong similarity. The effect of carotenoid mutations on the photosynthetic phenotype has been studied by examining growth rates, pigments, pigment-protein complexes and gene expression for a complete set of carotenoid mutants. 161 refs.

  19. Assignment of the sup 1 H and sup 15 N NMR spectra of Rhodobacter capsulatus ferrocytochrome c sub 2

    SciTech Connect

    Gooley, P.R.; Caffrey, M.S.; Cusanovich, M.A.; MacKenzie, N.E. )

    1990-03-06

    The peptide resonances of the {sup 1}H and {sup 15}N nuclear magnetic resonance spectra of ferrocytochrome c{sub 2} from Rhodobacter capsulatus are sequentially assigned by a combination of 2D {sup 1}H-{sup 1}H and {sup 1}H-{sup 15}N spectroscopy, the latter performed on {sup 15}N-enriched protein. Short-range nuclear Overhauser effect (NOE) data show {alpha}-helices from residues 3-17, 55-65, 69-88, and 103-115. Within the latter two {alpha}-helices, there are three single 3{sub 10} turns, 70-72, 76-78, and 107-109. In addition {alpha}H-NH{sub i+1} and {alpha}H-NH{sub i+2} NOEs indicate that the N-terminal helix (3-17) is distorted. Compared to horse or tuna cytochrome c and cytochrome c{sub 2} of Rhodospirillium rubrum, there is a 6-residue insertion at residues 23-29 in R. capsulatus cytochrome c{sub 2}. The NOE data show that this insertion forms a loop, probably an {Omega} loop. {sup 1}H-{sup 15}N heteronuclear multiple quantum correlation experiments are used to follow NH exchange over a period of 40 h. As the 2D spectra are acquired in short time periods (30 min), rates for intermediate exchanging protons can be measured. Comparison of the NH exchange data for the N-terminal helix of cytochrome c{sub 2} of R. capsulatus with the highly homologous horse heart cytochrome c shows that this helix is less stable in cytochrome c{sub 2}.

  20. Differential levels of specific cytochrome c biogenesis proteins in response to oxygen: analysis of the ccl operon in Rhodobacter capsulatus.

    PubMed Central

    Gabbert, K K; Goldman, B S; Kranz, R G

    1997-01-01

    The photosynthetic bacterium Rhodobacter capsulatus synthesizes c-type cytochromes under a variety of growth conditions. For example, under aerobic growth, c-type cytochromes are synthesized as part of an electron transport pathway, using oxygen as the terminal electron acceptor. Anaerobically in the light, R. capsulatus requires cytochrome bc1 and other c-type cytochromes for the photosynthetic electron transport pathway. It is shown here that the ccl1 and ccl2 genes of R. capsulatus are required for the synthesis of all c-type cytochromes, including the cytochrome c' protein of unknown function but of structural similarity to cytochrome b562. Polar and nonpolar mutations constructed in each gene demonstrated that the ccl12 genes form an operon. Expression of the ccl12 genes was examined by using lacZ and phoA fusions as translational reporters. Primer extension analysis was used to determine transcriptional control and the start site of the ccl12 promoter. Finally, antiserum to the Ccl2 protein was used to quantitate levels of Ccl2 under six different growth conditions. The Ccl2 protein is present at 20-fold-higher levels under conditions where oxygen is present. In contrast, other cytochromes c biogenesis proteins, HelA and HelX, previously shown to be part of an helABCDX operon, are at relatively similar levels under these six growth conditions. This discovery is discussed in terms of the physiology and evolution of cytochromes c biogenesis, with particular attention to oxidative environments. PMID:9286996

  1. Orbiting droplets on a vibrated bath

    NASA Astrophysics Data System (ADS)

    Sampara, Naresh; Burger, Loic; Gilet, Tristan; Microfluidics, university of liege Team

    2015-11-01

    A millimeter-sized oil droplet can bounce on a vertically vibrated liquid bath for unlimited time. It may couple to the surface wave it emits; leading to horizontal self-propulsion called walking. When several walkers coexist close to one another, they either repel or attract each other, in response to the superposition of the waves they generate. Attraction leads to various bound states, including droplets that orbit around each other. We have experimentally investigated the variety of quantized orbital motions exhibited by two, three and more identical walkers, as a function of forcing acceleration. Each motion is quantified in terms of droplet and wave energy.

  2. A Cu{sup 2+} site common to photosynthetic bacterial reaction centers from Rb. sphaeroides, Rb. capsulatus, and Rps. viridis.

    SciTech Connect

    Utschig, L. M.; Poluektov, O.; Schlesselman, S. L.; Thurnauer, M. C.; Tiede, D. M.; Chemistry

    2001-05-22

    The interaction of metal ions with isolated photosynthetic reaction centers (RCs) from the purple bacteria Rhodobacter sphaeroides, Rhodobacter capsulatus, and Rhodopseudomonas viridis has been investigated with transient optical and magnetic resonance techniques. In RCs from all species, the electrochromic response of the bacteriopheophytin cofactors associated with Q{sub A}{sup -}Q{sub B} {yields} Q{sub A}Q{sub B}{sup -} electron transfer is slowed in the presence of Cu{sup 2+}. This slowing is similar to the metal ion effect observed for RCs from Rb. sphaeroides where Zn{sup 2+} was bound to a specific site on the surface of the RC [Utschig et al. (1998) Biochemistry 37, 8278]. The coordination environments of the Cu{sup 2+} sites were probed with electron paramagnetic resonance (EPR) spectroscopy, providing the first direct spectroscopic evidence for the existence of a second metal site in RCs from Rb. capsulatus and Rps. viridis. In the dark, RCs with Cu{sup 2+} bound to the surface exhibit axially symmetric EPR spectra. Electron spin echo envelope modulation (ESEEM) spectral results indicate multiple weakly hyperfine coupled {sup 14}N nuclei in close proximity to Cu{sup 2+}. These ESEEM spectra resemble those observed for Cu{sup 2+} RCs from Rb. sphaeroides [Utschig et al. (2000) Biochemistry 39, 2961] and indicate that two or more histidines ligate the Cu{sup 2+} at the surface site in each RC. Thus, RCs from Rb. sphaeroides, Rb. capsulatus, and Rps. viridis each have a structurally analogous Cu{sup 2+} binding site that is involved in modulating the Q{sub A}{sup -}Q{sub B} {yields} Q{sub A}Q{sub B}{sup -} electron-transfer process. Inspection of the Rps. viridis crystal structure reveals four potential histidine ligands from three different subunits (M16, H178, H72, and L211) located beneath the Q{sub B} binding pocket. The location of these histidines is surprisingly similar to the grouping of four histidine residues (H68, H126, H128, and L211) observed in

  3. Assisting Cognitively Impaired Nursing Home Residents with Bathing: Effects of Two Bathing Interventions on Caregiving

    ERIC Educational Resources Information Center

    Hoeffer, Beverly; Talerico, Karen Amann; Rasin, Joyce; Mitchell, C. Madeline; Stewart, Babara J.; McKenzie, Darlene; Barrick, Ann Louise; Rader, Joanne; Sloane, Philip D.

    2006-01-01

    Purpose: When cognitively impaired nursing home residents exhibit agitated and aggressive behaviors during bathing, nursing home caregivers are in a unique position to improve residents' experience. This report addresses whether certified nursing assistants (CNAs) who received training in a person-centered approach with showering and with the…

  4. Carbon Isotope Fractionations Associated with Methanotrophic Growth with the Soluble and Particulate Methane Monooxygenases

    NASA Technical Reports Server (NTRS)

    Jahnke, Linda L.; Summons, Roger E.; Chang, Sherwood (Technical Monitor)

    1996-01-01

    Growth experiments with the RuMP-type methanotroph, Methylococcus capsulatus (Bath), have demonstrated that biomass and lipid biomarkers are significantly depleted in C-13 compared to the substrate methane and that the extent of fractionation is dependent on whether cells express the soluble (s) or particulate (p) methane monooxygenase (MMO). The presence or absence of the characteristic sMMO subunits was monitored using SDS-polyacrylamide gels. In M. capsulatus grown with no Cu supplementation, the characteristic sMMO subunits were observed in the soluble fraction throughout the entire growth period and biomass was depleted in C-13 by approximately 14,700 relative to substrate methane. In cells grown with 5uM Cu, no sMMO bands were observed and a greater fractionation of approximately 27,700 in resultant biomass was obtained. Methanol growth experiments with M. capsulatus and with a RuMP methylotroph, Methylophilus methylotrophus, in which biomass measurements yielded depletions in C-13 of 9 and 5%(sub o), respectively, suggest that oxidation of methane is the major fractionation step. Growth of M. capsulatus at a low level of oxygen, approximately 0.5%, had no significant effect on carbon isotope fractionation by either sMMO or pMMO. These observations are significant for identification of molecular biomarkers; and methanotrophic contributions to carbon isotope composition in natural environments.

  5. Recovery process for electroless plating baths

    DOEpatents

    Anderson, R.W.; Neff, W.A.

    1992-05-12

    A process is described for removing, from spent electroless metal plating bath solutions, accumulated byproducts and counter-ions that have deleterious effects on plating. The solution, or a portion thereof, is passed through a selected cation exchange resin bed in hydrogen form, the resin selected from strong acid cation exchangers and combinations of intermediate acid cation exchangers with strong acid cation exchangers. Sodium and nickel ions are sorbed in the selected cation exchanger, with little removal of other constituents. The remaining solution is subjected to sulfate removal through precipitation of calcium sulfate hemihydrate using, sequentially, CaO and then CaCO[sub 3]. Phosphite removal from the solution is accomplished by the addition of MgO to form magnesium phosphite trihydrate. The washed precipitates of these steps can be safely discarded in nontoxic land fills, or used in various chemical industries. Finally, any remaining solution can be concentrated, adjusted for pH, and be ready for reuse. The plating metal can be removed from the exchanger with sulfuric acid or with the filtrate from the magnesium phosphite precipitation forming a sulfate of the plating metal for reuse. The process is illustrated as applied to processing electroless nickel plating baths. 18 figs.

  6. Recovery process for electroless plating baths

    DOEpatents

    Anderson, Roger W.; Neff, Wayne A.

    1992-01-01

    A process for removing, from spent electroless metal plating bath solutions, accumulated byproducts and counter-ions that have deleterious effects on plating. The solution, or a portion thereof, is passed through a selected cation exchange resin bed in hydrogen form, the resin selected from strong acid cation exchangers and combinations of intermediate acid cation exchangers with strong acid cation exchangers. Sodium and nickel ions are sorbed in the selected cation exchanger, with little removal of other constituents. The remaining solution is subjected to sulfate removal through precipitation of calcium sulfate hemihydrate using, sequentially, CaO and then CaCO.sub.3. Phosphite removal from the solution is accomplished by the addition of MgO to form magnesium phosphite trihydrate. The washed precipitates of these steps can be safely discarded in nontoxic land fills, or used in various chemical industries. Finally, any remaining solution can be concentrated, adjusted for pH, and be ready for reuse. The plating metal can be removed from the exchanger with sulfuric acid or with the filtrate from the magnesium phosphite precipitation forming a sulfate of the plating metal for reuse. The process is illustrated as applied to processing electroless nickel plating baths.

  7. Cavity-assisted quantum bath engineering

    NASA Astrophysics Data System (ADS)

    Murch, Kater

    2013-03-01

    In practice, quantum systems are never completely isolated, but instead interact with degrees of freedom in the surrounding environment, eventually leading to decoherence. Precision measurement techniques such as nuclear magnetic resonance and interferometry, as well as envisioned quantum schemes for computation, simulation, and data encryption, rely on the ability to prepare and preserve delicate quantum superpositions and entanglement. The conventional route to long-lived quantum coherence involves minimizing coupling to a dissipative bath. Paradoxically, it is possible to instead engineer specific couplings to a quantum environment that allow dissipation to actually preserve coherence. I will discuss our recent demonstration of quantum bath engineering for a superconducting qubit coupled to a microwave cavity. By tailoring the spectrum of microwave photon shot noise in the cavity, we create a dissipative environment that autonomously relaxes the qubit to an arbitrarily specified coherent superposition of the ground and excited states. In the presence of background thermal excitations, this mechanism increases the state purity and effectively cools the dressed atom state to a low temperature. We envision that future multi-qubit implementations could enable the preparation of entangled many-body states suitable for quantum simulation and computation. This work was supported by the IARPA CSQ program.

  8. Cavitation effects in ultrasonic cleaning baths

    NASA Technical Reports Server (NTRS)

    Glasscock, Barbara H.

    1995-01-01

    In this project, the effect of cavitation from aqueous ultrasonic cleaning on the surfaces of metal and non-metal sample coupons was studied. After twenty cleaning cycles, the mass loss from the aluminum coupons averaged 0.22 mg/sq cm surface area and 0.014 mg/sq cm for both stainless steel and titanium. The aluminum coupons showed visual evidence of minor cavitation erosion in regions of previously existing surface irregularities. The non-metal samples showed some periods of mass gain. These effects are believed to have minor impact on hardware being cleaned, but should be evaluated in the context of specific hardware requirements. Also the ultrasonic activity in the large cleaning baths was found to be unevenly distributed as measured by damage to sheets of aluminum foil. It is therefore recommended that items being cleaned in an ultrasonic bath be moved or conveyed during the cleaning to more evenly distribute the cavitation action provide more uniform cleaning.

  9. Thermal baths as quantum resources: more friends than foes?

    NASA Astrophysics Data System (ADS)

    Kurizki, Gershon; Shahmoon, Ephraim; Zwick, Analia

    2015-12-01

    In this article we argue that thermal reservoirs (baths) are potentially useful resources in processes involving atoms interacting with quantized electromagnetic fields and their applications to quantum technologies. One may try to suppress the bath effects by means of dynamical control, but such control does not always yield the desired results. We wish instead to take advantage of bath effects, that do not obliterate ‘quantumness’ in the system-bath compound. To this end, three possible approaches have been pursued by us. (i) Control of a quantum system faster than the correlation time of the bath to which it couples: such control allows us to reveal quasi-reversible/coherent dynamical phenomena of quantum open systems, manifest by the quantum Zeno or anti-Zeno effects (QZE or AZE, respectively). Dynamical control methods based on the QZE are aimed not only at protecting the quantumness of the system, but also diagnosing the bath spectra or transferring quantum information via noisy media. By contrast, AZE-based control is useful for fast cooling of thermalized quantum systems. (ii) Engineering the coupling of quantum systems to selected bath modes: this approach, based on field-atom coupling control in cavities, waveguides and photonic band structures, allows one to drastically enhance the strength and range of atom-atom coupling through the mediation of the selected bath modes. More dramatically, it allows us to achieve bath-induced entanglement that may appear paradoxical if one takes the conventional view that coupling to baths destroys quantumness. (iii) Engineering baths with appropriate non-flat spectra: this approach is a prerequisite for the construction of the simplest and most efficient quantum heat machines (engines and refrigerators). We may thus conclude that often thermal baths are ‘more friends than foes’ in quantum technologies.

  10. Ultrasonic bath depth control and regulation in single cell recordings.

    PubMed

    Duong Dinh, Thien An; Jüngling, Eberhard; Strotmann, Karl-Heinz; Westhofen, Martin; Lückhoff, Andreas

    2006-09-01

    Control of the bath depth is critical in many applications of the patch-clamp technique, particularly when the capacitance of cells is determined to assess secretion or transmitter release or in studies of ion currents sensitive to small changes in the hydrostatic pressure. We describe an inexpensive technique for tight control of the bath depth with the aid of a commercially available ultrasound sensor. The sensor continuously determines changes in the distance to the bath surface with a resolution of about 10 mum. The signal from the sensor is digitized in a microcontroller card and used to send on or off signals at 100 Hz to a peristaltic pump that removes volume from the bath. The inflow into the bath can be realized in a versatile way. The capacitance of Sylgard-coated patch-clamp glass electrodes, demonstrated to be extremely sensitive to small changes in the area moistened by bath solution, is constant within the noise level of +/-3 fF when immersed into a depth-controlled bath, even during exchange of the bath medium. Thus, when small changes in the cell capacitance are measured in patch-clamp experiments, errors due to alterations in the pipette capacitance caused by bath depth fluctuations are eliminated. PMID:16721611

  11. An assessment of the impact of the proposed EU bathing water directive on Irish coastal bathing area compliance.

    PubMed

    Chawla, R; Real, K; Masterson, B

    2005-01-01

    An assessment of the impact of the new microbial water quality standards of the proposed EU Bathing Water Directive on the classification of designated Irish coastal bathing areas is presented. The new standards are applied retrospectively to the microbial water quality results for the bathing seasons of 1999, 2000 and 2001, and the outcome is compared with that recorded under the present Bathing Water Directive. A Microsoft EXCEL application was developed to generate the retrospective bathing area classifications according to the proposed Directive (Excellent, Good, Poor). It was found that the number of Irish coastal bathing areas not attaining 'Excellent' classification (as would be required at present for the Blue Flag award) was trebled; the number attaining 'Good' classification was increased by about 50%, and the number attracting 'Poor' classification (equivalent to 'Fail' under the present Directive) was increased nine-fold. Some of the shortcomings of the proposed Directive and suggestions for its revision are discussed. PMID:15850194

  12. Optimizing multi-step B-side charge separation in photosynthetic reaction centers from Rhodobacter capsulatus.

    PubMed

    Faries, Kaitlyn M; Kressel, Lucas L; Dylla, Nicholas P; Wander, Marc J; Hanson, Deborah K; Holten, Dewey; Laible, Philip D; Kirmaier, Christine

    2016-02-01

    Using high-throughput methods for mutagenesis, protein isolation and charge-separation functionality, we have assayed 40 Rhodobacter capsulatus reaction center (RC) mutants for their P(+)QB(-) yield (P is a dimer of bacteriochlorophylls and Q is a ubiquinone) as produced using the normally inactive B-side cofactors BB and HB (where B is a bacteriochlorophyll and H is a bacteriopheophytin). Two sets of mutants explore all possible residues at M131 (M polypeptide, native residue Val near HB) in tandem with either a fixed His or a fixed Asn at L181 (L polypeptide, native residue Phe near BB). A third set of mutants explores all possible residues at L181 with a fixed Glu at M131 that can form a hydrogen bond to HB. For each set of mutants, the results of a rapid millisecond screening assay that probes the yield of P(+)QB(-) are compared among that set and to the other mutants reported here or previously. For a subset of eight mutants, the rate constants and yields of the individual B-side electron transfer processes are determined via transient absorption measurements spanning 100 fs to 50 μs. The resulting ranking of mutants for their yield of P(+)QB(-) from ultrafast experiments is in good agreement with that obtained from the millisecond screening assay, further validating the efficient, high-throughput screen for B-side transmembrane charge separation. Results from mutants that individually show progress toward optimization of P(+)HB(-)→P(+)QB(-) electron transfer or initial P*→P(+)HB(-) conversion highlight unmet challenges of optimizing both processes simultaneously. PMID:26658355

  13. 30 CFR 75.1712 - Bath houses and toilet facilities.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 30 Mineral Resources 1 2014-07-01 2014-07-01 false Bath houses and toilet facilities. 75.1712 Section 75.1712 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR COAL MINE SAFETY AND HEALTH MANDATORY SAFETY STANDARDS-UNDERGROUND COAL MINES Miscellaneous § 75.1712 Bath...

  14. 30 CFR 75.1712 - Bath houses and toilet facilities.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 30 Mineral Resources 1 2013-07-01 2013-07-01 false Bath houses and toilet facilities. 75.1712 Section 75.1712 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR COAL MINE SAFETY AND HEALTH MANDATORY SAFETY STANDARDS-UNDERGROUND COAL MINES Miscellaneous § 75.1712 Bath...

  15. 30 CFR 75.1712 - Bath houses and toilet facilities.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 30 Mineral Resources 1 2012-07-01 2012-07-01 false Bath houses and toilet facilities. 75.1712 Section 75.1712 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR COAL MINE SAFETY AND HEALTH MANDATORY SAFETY STANDARDS-UNDERGROUND COAL MINES Miscellaneous § 75.1712 Bath...

  16. 30 CFR 75.1712 - Bath houses and toilet facilities.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 30 Mineral Resources 1 2011-07-01 2011-07-01 false Bath houses and toilet facilities. 75.1712 Section 75.1712 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR COAL MINE SAFETY AND HEALTH MANDATORY SAFETY STANDARDS-UNDERGROUND COAL MINES Miscellaneous § 75.1712 Bath...

  17. 30 CFR 75.1712 - Bath houses and toilet facilities.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Bath houses and toilet facilities. 75.1712 Section 75.1712 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR COAL MINE SAFETY AND HEALTH MANDATORY SAFETY STANDARDS-UNDERGROUND COAL MINES Miscellaneous § 75.1712 Bath...

  18. Chemical Safety: Molten Salt Baths Cited as Lab Hazards.

    ERIC Educational Resources Information Center

    Baum, Rudy

    1982-01-01

    Discusses danger of explosions with molten salts baths, commonly used as heat-transfer media. One such explosion involved use of a bath containing 3-lb sodium nitrite and 1-lb potassium thiocyanate. Although most commercially available mixtures for heat transfer contain oxidizers, a reducer (thiocyanate) was included which possibly triggered the…

  19. 20 CFR 654.412 - Bathing, laundry, and handwashing.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... facilities, supplied with hot and cold water under pressure, shall be provided for the use of all occupants... Bathing, laundry, and handwashing. (a) Bathing and handwashing facilities, supplied with hot and cold water under pressure, shall be provided for the use of all occupants. These facilities shall be...

  20. 36 CFR 21.11 - Redemption of bath tickets.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 36 Parks, Forests, and Public Property 1 2010-07-01 2010-07-01 false Redemption of bath tickets. 21.11 Section 21.11 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR HOT SPRINGS NATIONAL PARK; BATHHOUSE REGULATIONS § 21.11 Redemption of bath tickets....

  1. 36 CFR 21.11 - Redemption of bath tickets.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 36 Parks, Forests, and Public Property 1 2011-07-01 2011-07-01 false Redemption of bath tickets. 21.11 Section 21.11 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR HOT SPRINGS NATIONAL PARK; BATHHOUSE REGULATIONS § 21.11 Redemption of bath tickets....

  2. 36 CFR 21.12 - Lost bath tickets.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 36 Parks, Forests, and Public Property 1 2012-07-01 2012-07-01 false Lost bath tickets. 21.12 Section 21.12 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR HOT SPRINGS NATIONAL PARK; BATHHOUSE REGULATIONS § 21.12 Lost bath tickets. A patron who loses his ticket...

  3. 36 CFR 21.12 - Lost bath tickets.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 36 Parks, Forests, and Public Property 1 2010-07-01 2010-07-01 false Lost bath tickets. 21.12 Section 21.12 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR HOT SPRINGS NATIONAL PARK; BATHHOUSE REGULATIONS § 21.12 Lost bath tickets. A patron who loses his ticket...

  4. 36 CFR 21.5 - Therapeutic bathing requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 36 Parks, Forests, and Public Property 1 2010-07-01 2010-07-01 false Therapeutic bathing requirements. 21.5 Section 21.5 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR HOT SPRINGS NATIONAL PARK; BATHHOUSE REGULATIONS § 21.5 Therapeutic bathing requirements....

  5. 36 CFR 21.11 - Redemption of bath tickets.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 36 Parks, Forests, and Public Property 1 2012-07-01 2012-07-01 false Redemption of bath tickets. 21.11 Section 21.11 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR HOT SPRINGS NATIONAL PARK; BATHHOUSE REGULATIONS § 21.11 Redemption of bath tickets....

  6. 36 CFR 21.11 - Redemption of bath tickets.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 36 Parks, Forests, and Public Property 1 2013-07-01 2013-07-01 false Redemption of bath tickets. 21.11 Section 21.11 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR HOT SPRINGS NATIONAL PARK; BATHHOUSE REGULATIONS § 21.11 Redemption of bath tickets....

  7. 36 CFR 21.5 - Therapeutic bathing requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 36 Parks, Forests, and Public Property 1 2011-07-01 2011-07-01 false Therapeutic bathing requirements. 21.5 Section 21.5 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR HOT SPRINGS NATIONAL PARK; BATHHOUSE REGULATIONS § 21.5 Therapeutic bathing requirements....

  8. 36 CFR 21.5 - Therapeutic bathing requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 36 Parks, Forests, and Public Property 1 2012-07-01 2012-07-01 false Therapeutic bathing requirements. 21.5 Section 21.5 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR HOT SPRINGS NATIONAL PARK; BATHHOUSE REGULATIONS § 21.5 Therapeutic bathing requirements....

  9. 36 CFR 21.5 - Therapeutic bathing requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 36 Parks, Forests, and Public Property 1 2013-07-01 2013-07-01 false Therapeutic bathing requirements. 21.5 Section 21.5 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR HOT SPRINGS NATIONAL PARK; BATHHOUSE REGULATIONS § 21.5 Therapeutic bathing requirements....

  10. 36 CFR 21.12 - Lost bath tickets.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 36 Parks, Forests, and Public Property 1 2011-07-01 2011-07-01 false Lost bath tickets. 21.12 Section 21.12 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR HOT SPRINGS NATIONAL PARK; BATHHOUSE REGULATIONS § 21.12 Lost bath tickets. A patron who loses his ticket...

  11. 36 CFR 21.12 - Lost bath tickets.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 36 Parks, Forests, and Public Property 1 2013-07-01 2013-07-01 false Lost bath tickets. 21.12 Section 21.12 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR HOT SPRINGS NATIONAL PARK; BATHHOUSE REGULATIONS § 21.12 Lost bath tickets. A patron who loses his ticket...

  12. Processing a printed wiring board by single bath electrodeposition

    DOEpatents

    Meltzer, Michael P.; Steffani, Christopher P.; Gonfiotti, Ray A.

    2010-12-07

    A method of processing a printed wiring board. Initial processing steps are implemented on the printed wiring board. Copper is plated on the printed wiring board from a bath containing nickel and copper. Nickel is plated on the printed wiring board from a bath containing nickel and copper and final processing steps are implemented on the printed wiring board.

  13. 21 CFR 740.17 - Foaming detergent bath products.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 7 2010-04-01 2010-04-01 false Foaming detergent bath products. 740.17 Section 740.17 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) COSMETICS COSMETIC PRODUCT WARNING STATEMENTS Warning Statements § 740.17 Foaming detergent bath...

  14. 21 CFR 740.17 - Foaming detergent bath products.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 7 2011-04-01 2010-04-01 true Foaming detergent bath products. 740.17 Section 740.17 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) COSMETICS COSMETIC PRODUCT WARNING STATEMENTS Warning Statements § 740.17 Foaming detergent bath...

  15. 21 CFR 740.17 - Foaming detergent bath products.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 7 2012-04-01 2012-04-01 false Foaming detergent bath products. 740.17 Section 740.17 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) COSMETICS COSMETIC PRODUCT WARNING STATEMENTS Warning Statements § 740.17 Foaming detergent bath...

  16. 21 CFR 740.17 - Foaming detergent bath products.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 7 2014-04-01 2014-04-01 false Foaming detergent bath products. 740.17 Section 740.17 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) COSMETICS COSMETIC PRODUCT WARNING STATEMENTS Warning Statements § 740.17 Foaming detergent bath...

  17. 21 CFR 740.17 - Foaming detergent bath products.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 7 2013-04-01 2013-04-01 false Foaming detergent bath products. 740.17 Section 740.17 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) COSMETICS COSMETIC PRODUCT WARNING STATEMENTS Warning Statements § 740.17 Foaming detergent bath...

  18. Copper Sulfate Foot Baths on Dairies and Crop Toxicities

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A rising concern with the application of dairy wastes to agricultural fields is the accumulation of copper (Cu) in the soil. Copper sulfate (CuSO4) from cattle foot baths are washed out of dairy barns and into wastewater lagoons. The addition of CuSO4 baths has been reported to increase Cu concent...

  19. 20 CFR 654.412 - Bathing, laundry, and handwashing.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... Bathing, laundry, and handwashing. (a) Bathing and handwashing facilities, supplied with hot and cold... of floor space per unit. Adequate, dry dressing space shall be provided in common use facilities.... When common use shower facilities for both sexes are in the same building they shall be separated by...

  20. 20 CFR 654.412 - Bathing, laundry, and handwashing.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 20 Employees' Benefits 3 2012-04-01 2012-04-01 false Bathing, laundry, and handwashing. 654.412 Section 654.412 Employees' Benefits EMPLOYMENT AND TRAINING ADMINISTRATION, DEPARTMENT OF LABOR SPECIAL RESPONSIBILITIES OF THE EMPLOYMENT SERVICE SYSTEM Housing for Agricultural Workers Housing Standards § 654.412 Bathing, laundry, and handwashing....

  1. 36 CFR 21.5 - Therapeutic bathing requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 36 Parks, Forests, and Public Property 1 2014-07-01 2014-07-01 false Therapeutic bathing requirements. 21.5 Section 21.5 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR HOT SPRINGS NATIONAL PARK; BATHHOUSE REGULATIONS § 21.5 Therapeutic bathing requirements....

  2. 36 CFR 21.12 - Lost bath tickets.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 36 Parks, Forests, and Public Property 1 2014-07-01 2014-07-01 false Lost bath tickets. 21.12 Section 21.12 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR HOT SPRINGS NATIONAL PARK; BATHHOUSE REGULATIONS § 21.12 Lost bath tickets. A patron who loses his ticket...

  3. 36 CFR 21.11 - Redemption of bath tickets.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 36 Parks, Forests, and Public Property 1 2014-07-01 2014-07-01 false Redemption of bath tickets. 21.11 Section 21.11 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR HOT SPRINGS NATIONAL PARK; BATHHOUSE REGULATIONS § 21.11 Redemption of bath tickets....

  4. The development of a virtual heat bath for calorimeters

    SciTech Connect

    Pickrell, M.M.; Bracken, D.S.; Rudy, C.R.

    1998-12-31

    All existing calorimeter systems for sensitive nuclear assay employ a heat bath surrounding the sample chamber. The purpose of the heat bath is to maintain a constant temperature so that a fixed temperature difference is maintained across the thermal resistance of the calorimeter. Present calorimeter systems all employ an active, feedback-controlled system to maintain a fixed temperature. An alternative would be to allow the heat-bath temperature to change, to measure it, and to compensate the assay for this change. Two significant observations make this approach possible: (1) the effect on the measurement of a temperature change in the heat bath is differential in form and (2) temperature measurement systems are very accurate when measuring differences in temperature (either in time or between two locations). From these observations, the authors have developed a virtual heat-bath compensation system. The control theory and results will be presented.

  5. Photo-biohydrogen production potential of Rhodobacter capsulatus-PK from wheat straw

    PubMed Central

    2013-01-01

    Background Biotechnological exploitation of lignocellulosic biomass is promising for sustainable and environmentally sound energy provision strategy because of the abundant availability of the renewable resources. Wheat straw (WS) comprising of 75-80% cellulose and hemicellulose is one of widely available, inexpensive and renewable lignocellulosic biomass types. The cellulosic and hemicellulose substrate can be hydrolyzed into monomeric sugars by chemical and/or biological methods. Results This study examined comparative potential of dilute acid and pre-ammonia pretreated and enzymatically hydrolyzed wheat straw (WS) for hydrogen production by purple non sulfur bacterium Rhodobacter capsulatus-PK. Gas production became noticeable after 14 h of inoculation in WS pretreated with 4% H2SO4. The detoxified liquid hydrolyzate (DLH) after overliming attained a production level of 372 mL-H2/L after 16 h under illumination of 120-150 W/m2 at 30 ± 2.0°C. Whereas the non-detoxified acid pretreated hydrolyzate (NDLH) of WS could produce only upto 254 mL-H2/L after 21 h post inoculation. Evolution of H2 became observable just after 10 ± 2.0 h of inoculation by employing 48 h age inoculum on the WS pretreated with 30% ammonia, hydrolyzed with cellulase 80 FPU/g and β-glucosidase 220 CbU/ml at 50°C. Upto 712 ml/L of culture was measured with continuous shaking for 24 h. The 47.5% and 64.2% higher hydrogen volume than the DLH and NDLH substrates, respectively appeared as a function of significantly higher monomeric sugar contents of the enzymatically hydrolyzed substrate and lesser/zero amounts of toxic derivatives including pH reducing agents. Conclusion Photofermentative hydrogen production from lignocellulosic waste is a feasible approach for eco-friendly sustainable supply of bioenergy in a cost-effective way. Results of this study provide new insight for addressing biotechnological exploitation of abundantly available and low-cost cellulosic substrates

  6. Bath for electrolytic reduction of alumina and method therefor

    DOEpatents

    Brown, Craig W.; Brooks, Richard J.; Frizzle, Patrick B.; Juric, Drago D.

    2002-11-26

    An electrolytic bath for use during the electrolytic reduction of alumina to aluminum. The bath comprises a molten electrolyte having the following ingredients: (a) AlF.sub.3 and at least one salt selected from the group consisting of NaF, KF, and LiF; and (b) about 0.004 wt. % to about 0.2 wt. %, based on total weight of the molten electrolyte, of at least one transition metal or at least one compound of the metal or both. The compound may be, for example, a fluoride, oxide, or carbonate. The metal can be nickel, iron, copper, cobalt, or molybdenum. The bath can be employed in a combination that includes a vessel for containing the bath and at least one non-consumable anode and at least one dimensionally stable cathode in the bath. Employing the bath of the present invention during electrolytic reduction of alumina to aluminum can improve the wetting of aluminum on a cathode by reducing or eliminating the formation of non-metallic deposits on the cathode. Removing sulfur from the bath can also minimize cathode deposits. Aluminum formed on the cathode can be removed directly from the cathode.

  7. Mephedrone ("bath salt") pharmacology: insights from invertebrates.

    PubMed

    Ramoz, L; Lodi, S; Bhatt, P; Reitz, A B; Tallarida, C; Tallarida, R J; Raffa, R B; Rawls, S M

    2012-04-19

    Psychoactive bath salts (also called meph, drone, meow meow, m-CAT, bounce, bubbles, mad cow, etc.) contain a substance called mephedrone (4-methylcathinone) that may share psychostimulant properties with amphetamine and cocaine. However, there are only limited studies of the neuropharmacological profile of mephedrone. The present study used an established invertebrate (planarian) assay to test the hypothesis that acute and repeated mephedrone exposure produces psychostimulant-like behavioral effects. Acute mephedrone administration (50-1000 μM) produced stereotyped movements that were attenuated by a dopamine receptor antagonist (SCH 23390) (0.3 μM). Spontaneous discontinuation of mephedrone exposure (1, 10 μM) (60 min) resulted in an abstinence-induced withdrawal response (i.e. reduced motility). In place conditioning experiments, planarians in which mephedrone (100, 500 μM) was paired with the non-preferred environment during conditioning displayed a shift in preference upon subsequent testing. These results suggest that mephedrone produces three behavioral effects associated with psychostimulant drugs, namely dopamine-sensitive stereotyped movements, abstinence-induced withdrawal, and environmental place conditioning. PMID:22300981

  8. Balamuthia mandrillaris therapeutic mud bath in Jamaica.

    PubMed

    Todd, C D; Reyes-Batlle, M; Piñero, J E; Martínez-Carretero, E; Valladares, B; Lindo, J F; Lorenzo-Morales, J

    2015-07-01

    Balamuthia mandrillaris is an emerging cause of encephalitis in humans. The transmission dynamics are poorly understood due to the high fatality rate and the sporadic nature of cases. Seventy-two soil samples were collected from beaches and the banks of lagoons, rivers, ponds, mineral springs and streams from across Jamaica and assayed for the presence of B. mandrillaris. Seventy-nine sites were sampled and the mitochondrial 16S rDNA gene of B. mandrillaris was amplified and sequenced to confirm the presence of the amoeba. One isolate of B. mandrillaris was recovered from soil from mineral spring which hosts an informal therapeutic mud bath business. Although B. mandrillaris is less frequently isolated from soil than other free-living amoebae, rubbing mud containing the organism onto the skin increases the likelihood of exposure and infection. This first report on the isolation of B. mandrillaris in the Caribbean and its presence in soil where human contact is likely warrants further investigation using serological methods to elucidate exposure patterns. PMID:25335452

  9. Hyperthermic effects of hand bathing: benefits of incorporating finger flexion-extension exercise

    PubMed Central

    Ohshige, Tadasu; Ohwatashi, Akihiko; Kiyama, Ryoji; Nishi, Hiroaki; Takamori, Akihisa

    2015-01-01

    [Purpose] The purpose of our study was to compare the effects of hand bathing using plain water and water supplemented with inorganic salt and carbonated gas and to assess the hyperthermic effects of performing finger flexion-extension exercise while bathing in water with carbonated gas and inorganic salt and without water. [Subjects and Methods] Sixteen healthy, young males were subjected to plain water bathing, CO2 bathing, kineto-CO2 bathing, or no bathing. CO2 bathing involved bathing in a solution of artificial bath additives including inorganic salts and carbon dioxide. Partial bathing of the hand was implemented for 20 minutes at 41 °C. The concentration of carbonic gas was set at 33 ppm. In the kineto-CO2 bathing condition, finger flexion-extension exercise was performed at 60 laps per minute in the same solution used in CO2 bathing. The control group engaged in the same exercise as those in the kineto-CO2 bathing group, but without bathing. [Results] A significant increase in deep-body temperature was observed in the CO2 bathing and kineto-CO2 bathing conditions compared with both the plain water bathing and control condition. [Conclusion] Significantly heightened hyperthermic effects were observed when finger flexion-extension exercise was performed during CO2 bathing. PMID:26834351

  10. Quantum Spin Baths Induced Transition of Decoherence and Entanglement

    SciTech Connect

    Chen Pochung; Lai Chengyan; Hung, J.-T.; Mou Chungyu

    2008-11-07

    We investigate the reduced dynamics of single or two qubits coupled to an interacting quantum spin bath modeled by a XXZ spin chain. By using the method of time-dependent density matrix renormalization group (t-DMRG), we evaluate nonperturbatively the induced decoherence and entanglement. We find that the behavior of both decoherence and entanglement strongly depend on the phase of the underlying spin bath. We show that spin baths can induce entanglement for an initially disentangled pair of qubits. We observe that entanglement sudden death only occurs in paramagnetic phase and discuss the effect of the coupling range.

  11. Bath additives for the treatment of childhood eczema (BATHE): protocol for multicentre parallel group randomised trial

    PubMed Central

    Santer, Miriam; Rumsby, Kate; Ridd, Matthew J; Francis, Nick A; Stuart, Beth; Chorozoglou, Maria; Wood, Wendy; Roberts, Amanda; Thomas, Kim S; Williams, Hywel C; Little, Paul

    2015-01-01

    Introduction Bath emollients are widely prescribed for childhood eczema, yet evidence of their benefits over direct application of emollients is lacking. Objectives To determine the clinical and cost-effectiveness of adding bath emollient to the standard management of eczema in children Methods and analysis Design: Pragmatic open 2-armed parallel group randomised controlled trial. Setting: General practitioner (GP) practices in England and Wales. Participants: Children aged over 12 months and less than 12 years with eczema, excluding inactive or very mild eczema (5 or less on Nottingham Eczema Severity Scale). Interventions: Children will be randomised to either bath emollients plus standard eczema care or standard eczema care only. Outcome measures: Primary outcome is long-term eczema severity, measured by the Patient-Oriented Eczema Measure (POEM) repeated weekly for 16 weeks. Secondary outcomes include: number of eczema exacerbations resulting in healthcare consultations over 1 year; eczema severity over 1 year; disease-specific and generic quality of life; medication use and healthcare resource use; cost-effectiveness. Aiming to detect a mean difference between groups of 2.0 (SD 7.0) in weekly POEM scores over 16 weeks (significance 0.05, power 0.9), allowing for 20% loss to follow-up, gives a total sample size of 423 children. We will use repeated measures analysis of covariance, or a mixed model, to analyse weekly POEM scores. We will control for possible confounders, including baseline eczema severity and child's age. Cost-effectiveness analysis will be carried out from a National Health Service (NHS) perspective. Ethics and dissemination This protocol was approved by Newcastle and North Tyneside 1 NRES committee 14/NE/0098. Follow-up will be completed in 2017. Findings will be disseminated to participants and carers, the public, dermatology and primary care journals, guideline developers and decision-makers. Trial registration number ISRCTN

  12. Structural and Phylogenetic Analysis of Rhodobacter capsulatus NifF: Uncovering General Features of Nitrogen-fixation (nif)-Flavodoxins

    PubMed Central

    Pérez-Dorado, Inmaculada; Bortolotti, Ana; Cortez, Néstor; Hermoso, Juan A.

    2013-01-01

    Analysis of the crystal structure of NifF from Rhodobacter capsulatus and its homologues reported so far reflects the existence of unique structural features in nif flavodoxins: a leucine at the re face of the isoalloxazine, an eight-residue insertion at the C-terminus of the 50’s loop and a remarkable difference in the electrostatic potential surface with respect to non-nif flavodoxins. A phylogenetic study on 64 sequences from 52 bacterial species revealed four clusters, including different functional prototypes, correlating the previously defined as “short-chain” with the firmicutes flavodoxins and the “long-chain” with gram-negative species. The comparison of Rhodobacter NifF structure with other bacterial flavodoxin prototypes discloses the concurrence of specific features of these functional electron donors to nitrogenase. PMID:23303276

  13. Evidence for a Regulatory Link of Nitrogen Fixation and Photosynthesis in Rhodobacter capsulatus via HvrA

    PubMed Central

    Kern, Monika; Kamp, Paul-Bertram; Paschen, Annette; Masepohl, Bernd; Klipp, Werner

    1998-01-01

    A Rhodobacter capsulatus reporter strain, carrying a constitutively expressed nifA gene and a nifH-lacZ gene fusion, was used for random transposon Tn5 mutagenesis to search for genes required for the NtrC-independent ammonium repression of NifA activity. A mutation in hvrA, which is known to be involved in low-light activation of the photosynthetic apparatus, released both ammonium and oxygen control of nifH expression in this reporter strain, demonstrating a regulatory link of nitrogen fixation and photosynthesis via HvrA. In addition, a significant increase in bacteriochlorophyll a (BChla) content was found in cells under nitrogen-fixing conditions. HvrA was not involved in this up-regulation of BChla. Instead, the presence of active nitrogenase seemed to be sufficient for this process, since no increase in BChla content was observed in different nif mutants. PMID:9537402

  14. Magnetic susceptibility tensor and heme contact shifts determinations in the Rhodobacter capsulatus ferricytochrome c': NMR and magnetic susceptibility studies.

    PubMed

    Tsan, P; Caffrey, M; Daku, M L; Cusanovich, M; Marion, D; Gans, P

    2001-03-14

    The 1H and 15N resonances of the carbon monoxide complex of ferrocytochrome c' of Rhodobacter capsulatus, a ferrous diamagnetic heme protein, have been extensively assigned by TOCSY-HSQC, NOESY-HSQC, and HSQC-NOESY-HSQC 3D heteronuclear experiments performed on a 7 mM sample labeled with 15N. Based on short-range and medium-range NOEs and H(N) exchange rates, the secondary structure consists of four helices: helix 1 (3-29), helix 2 (33-48), helix 3 (78-101), and helix 4 (103-125). The 15N, 1HN, and 1H(alpha) chemical shifts of the CO complex form are compared to those of the previously assigned oxidized (or ferric) state. From the chemical shift differences between these redox states, the orientation and the anisotropy of the paramagnetic susceptibility tensor have been determined using the crystallographic coordinates of the ferric state. The chi-tensor is axial, and the orientation of the z-axis is approximately perpendicular to the heme plane. The paramagnetic chemical shifts of the protons of the heme ligand have been determined and decomposed into the Fermi shift and dipolar shift contributions. Magnetic susceptibility studies in frozen solutions have been performed. Fits of the susceptibility data using the model of Maltempo (Maltempo, M. M. J. Chem. Phys. 1974, 61, 2540-2547) are consistent with a rather low contribution of the S = 3/2 spin state over the range of temperatures and confirm the value of the axial anisotropy. Values in the range 10.4-12.5 cm(-1) have been inferred for the axial zero-field splitting parameter (D). Analysis of the contact shift and the susceptibility data suggests that cytochrome c' of Rb. capsulatus exhibits a predominant high-spin character of the iron in the oxidized state at room temperature. PMID:11456869

  15. Urea Utilization in the Phototrophic Bacterium Rhodobacter capsulatus Is Regulated by the Transcriptional Activator NtrC

    PubMed Central

    Masepohl, Bernd; Kaiser, Björn; Isakovic, Nazila; Richard, Cynthia L.; Kranz, Robert G.; Klipp, Werner

    2001-01-01

    The phototrophic nonsulfur purple bacterium Rhodobacter capsulatus can use urea as a sole source of nitrogen. Three transposon Tn5-induced mutations (Xan-9, Xan-10, and Xan-19), which led to a Ure− phenotype, were mapped to the ureF and ureC genes, whereas two other Tn5 insertions (Xan-20 and Xan-22) were located within the ntrC and ntrB genes, respectively. As in Klebsiella aerogenes and other bacteria, the genes encoding urease (ureABC) and the genes required for assembly of the nickel metallocenter (ureD and ureEFG) are clustered in R. capsulatus (ureDABC-orf136-ureEFG). No homologues of Orf136 were found in the databases, and mutational analysis demonstrated that orf136 is not essential for urease activity or growth on urea. Analysis of a ureDA-lacZ fusion showed that maximum expression of the ure genes occurred under nitrogen-limiting conditions (e.g., serine or urea as the sole nitrogen source), but ure gene expression was not substrate (urea) inducible. Expression of the ure genes was strictly dependent on NtrC, whereas ς54 was not essential for urease activity. Expression of the ure genes was lower (by a factor of 3.5) in the presence of ammonium than under nitrogen-limiting conditions, but significant transcription was also observed in the presence of ammonium, approximately 10-fold higher than in an ntrC mutant background. Thus, ure gene expression in the presence of ammonium also requires NtrC. Footprint analyses demonstrated binding of NtrC to tandem binding sites upstream of the ureD promoter. Phosphorylation of NtrC increased DNA binding by at least eightfold. Although urea is effectively used as a nitrogen source in an NtrC-dependent manner, nitrogenase activity was not repressed by urea. PMID:11133958

  16. Interior view of bath room 05 with original toilet stall, ...

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

    Interior view of bath room 0-5 with original toilet stall, marble surround, and urinal, facing west. - Marine Barracks, Panama Canal, Barracks Building, 100' North of Thatcher Highway, Balboa, Former Panama Canal Zone, CZ

  17. 14. STEAM CABINETS & SITZ BATH IN STEAM ROOM. ...

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

    14. STEAM CABINETS & SITZ BATH IN STEAM ROOM. - Hot Springs National Park, Bathhouse Row, Fordyce Bathhouse: Mechanical & Piping Systems, State Highway 7, 1 mile north of U.S. Highway 70, Hot Springs, Garland County, AR

  18. 1. NORTHWEST FRONT, SOUTHWEST SIDE (SPRING HOUSE IN FOREGROUND; BATH ...

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

    1. NORTHWEST FRONT, SOUTHWEST SIDE (SPRING HOUSE IN FOREGROUND; BATH HOUSE AT REAR) (4 x 5 negative; 5 x 7 print) - Salt Sulphur Springs, Spring House, U.S. Route 219, Salt Sulphur Springs, Monroe County, WV

  19. Online process control of acidic texturisation baths with ion chromatography.

    PubMed

    Zimmer, Martin; Oltersdorf, Antje; Rentsch, Jochen

    2009-12-15

    Etching of silicon with mixtures of hydrofluoric acid and nitric acid is a widely used process in silicon solar cell fabrication. One precondition for an optimized usage of the acidic etching baths is the exact knowledge of the chemical bath composition. In this paper, we investigated a fast and online-capable method for the total analysis of all bath constituents by ion chromatography. The chromatographical system consists of a low-volume injection valve, which injects the concentrated samples directly into the KOH-based eluent. After separation and detection of nitrate and fluoride, a post-column derivatization with sodium molybdate is applied to detect the hexafluorosilicic acid, which enriches in the texturisation bath during the etching process. The results of the presented approach are discussed and compared with already published chromatographical and titration methods found in literature. PMID:19836511

  20. 11. GENERAL VIEW OF MEN'S BATH HALL. Hot Springs ...

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

    11. GENERAL VIEW OF MEN'S BATH HALL. - Hot Springs National Park, Bathhouse Row, Fordyce Bathhouse: Mechanical & Piping Systems, State Highway 7, 1 mile north of U.S. Highway 70, Hot Springs, Garland County, AR

  1. WORKER REMOVING SLAG FROM THE MOLTEN METAL BATH IN THE ...

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

    WORKER REMOVING SLAG FROM THE MOLTEN METAL BATH IN THE ELECTRIC FURNACE AFTER ADDING A CHEMICAL COAGULANT TO FORCE IT TO THE SURFACE. - Southern Ductile Casting Company, Melting, 2217 Carolina Avenue, Bessemer, Jefferson County, AL

  2. Thermal bath of Dirac field in non-inertial frames

    NASA Astrophysics Data System (ADS)

    Zhang, Anwei

    2016-07-01

    The properties of inertial Dirac thermal bath viewed from the perspective of a uniformly accelerated observer is investigated. Using the single-mode approximation, we obtain the expression of the thermal reduced density matrix, which is constructed by the effective partition function in another way compared with the case of scalar field. We deduce the spectrum of particles and antiparticles seen by Rob and antiRob, respectively. It is found that the fermionic thermal bath will suppress the appearance of Unruh particles.

  3. Intranasal substituted cathinone "bath salts" psychosis potentially exacerbated by diphenhydramine.

    PubMed

    Gunderson, Erik W; Kirkpatrick, Matthew G; Willing, Laura M; Holstege, Christopher P

    2013-01-01

    In this report, we describe a case of intranasal "bath salts"-associated psychosis. Symptoms developed during a 3-week binge and were potentially exacerbated by oral diphenhydramine taken for insomnia. The clinical case conference includes expert discussion from 3 disciplines: emergency medicine toxicology, behavioral pharmacology, and addiction medicine. It is hoped that the discussion will provide insight into the clinical aspects and challenges of addressing acute substituted cathinone toxicity, including acute psychosis, a major adverse effect of bath salts consumption. PMID:23732955

  4. RNase E Enzymes from Rhodobacter capsulatus and Escherichia coli Differ in Context- and Sequence-Dependent In Vivo Cleavage within the Polycistronic puf mRNA

    PubMed Central

    Heck, Claudia; Evguenieva-Hackenberg, Elena; Balzer, Angelika; Klug, Gabriele

    1999-01-01

    The 5′ pufQ mRNA segment and the pufLMX mRNA segment of Rhodobacter capsulatus exhibit different stabilities. Degradation of both mRNA segments is initiated by RNase E-mediated endonucleolytic cleavage. While Rhodobacter RNase E does not discriminate between the different sequences present around the cleavage sites within pufQ and pufL, Escherichia coli RNase E shows preference for the sequence harboring more A and U residues. PMID:10601223

  5. Bath for electrolytic reduction of alumina and method therefor

    DOEpatents

    Brown, Craig W.; Brooks, Richard J.; Frizzle, Patrick B.; Juric, Drago D.

    2001-07-10

    An electrolytic bath for use during the electrolytic reduction of alumina to aluminum. The bath comprises a molten electrolyte having the following ingredients: (a) AlF.sub.3 and at least one salt selected from the group consisting of NaF, KF, and LiF; and (b) about 0.004 wt. % to about 0.2 wt. %, based on total weight of the molten electrolyte, of at least one transition metal or at least one compound of the metal or both. The compound may be, for example, a fluoride, oxide, or carbonate. The metal can be nickel, iron, copper, cobalt, or molybdenum. The bath can be employed in a combination that includes a vessel for containing the bath and at least one non-consumable anode and at least one dimensionally stable cathode in the bath. Employing the bath of the present invention during electrolytic reduction of alumina to aluminum can improve the wetting of aluminum on a cathode by reducing or eliminating the formation of non-metallic deposits on the cathode.

  6. Effect of hyperthermic water bath on parameters of cellular immunity.

    PubMed

    Blazícková, S; Rovenský, J; Koska, J; Vigas, M

    2000-01-01

    Effects of hyperthermic water bath on selected immune parameters (lymphocyte subpopulations, natural killer (NK) cell counts and their activity) were studied in a group of 10 volunteers. Application of hyperthermic water bath (both topical and whole-body) was followed by a significant reduction of relative B lymphocyte counts. Whole-body hyperthermic water bath reduced relative total T lymphocyte counts, increased relative CD8+ T lymphocyte and NK cell counts and increased NK activity. Whole-body hyperthermic bath increased somatotropic hormone (STH) activity in eight out of 10 volunteers; higher relative counts of CD8+ lymphocytes and NK cells were observed compared with the group of volunteers not responding to hyperthermic water bath by STH secretion. In five volunteers STH was released in response to local hyperthermic water bath and the NK activity of lymphocytes also increased but their relative counts did not. The results suggest that these increases in CD8+ lymphocyte and NK cell counts are probably dependent on increased STH production. PMID:11146901

  7. ELECTRODIALYSIS AS A TECHNIQUE FOR EXTENDING ELECTROLESS NICKEL BATH LIFE-IMPROVING SELECTIVITY AND REDUCING LOSSES OF VALUABLE BATH COMPONENTS

    EPA Science Inventory

    Over the last decade electrodialysis has emerged as an effective technique for removing accumulated reactant counterions (sodium and sulfate) and reaction products (orthophosphite) that interfere with the electroless nickel plating process, thus extending bath life by up to 50 me...

  8. 78 FR 53734 - Proposed Extension of Approval of Information Collection; Comment Request-Infant Bath Seats

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-30

    ... infant bath seats. 75 FR 31691. On July 31, 2012, the Commission adopted the revised ASTM standard for infant bath seats, ASTM F1967-11a. 77 FR 45242. The requirements for infant bath seats are set forth... COMMISSION Proposed Extension of Approval of Information Collection; Comment Request--Infant Bath...

  9. Nucleotide sequence and genetic analysis of the Rhodobacter capsulatus ORF6-nifUI SVW gene region: possible role of NifW in homocitrate processing.

    PubMed

    Masepohl, B; Angermüller, S; Hennecke, S; Hübner, P; Moreno-Vivian, C; Klipp, W

    1993-04-01

    DNA sequence analysis of a 3494-bp HindIII-BclI fragment of the Rhodobacter capsulatus nif region A revealed genes that are homologous to ORF6, nifU, nifS, nifV and nifW from Azotobacter vinelandii and Klebsiella pneumoniae. R. capsulatus nifU, which is present in two copies, encodes a novel type of NifU protein. The deduced amino acid sequences of NifUI and NifUII share homology only with the C-terminal domain of NifU from A. vinelandii and K. pneumoniae. In contrast to nifA and nifB, which are almost perfectly duplicated, the predicted amino acid sequences of the two NifU proteins showed only 39% sequence identity. Expression of the ORF6-nifUISVW operon, which is preceded by a putative sigma 54-dependent promoter, required the function of NifA and the nif-specific rpoN gene product encoded by nifR4. Analysis of defined insertion and deletion mutants demonstrated that only nifS was absolutely essential for nitrogen fixation in R. capsulatus. Strains carrying mutations in nifV were capable of very slow diazotrophic growth, whereas ORF6, nifUI and nifW mutants as well as a nifUI/nifUII double mutant exhibited a Nif+ phenotype. Interestingly, R. capsulatus nifV mutants were able to reduce acetylene not only to ethylene but also to ethane under conditions preventing the expression of the alternative nitrogenase system. Homocitrate added to the growth medium repressed ethane formation and cured the NifV phenotype in R. capsulatus. Higher concentrations of homocitrate were necessary to complement the NifV phenotype of a polar nifV mutant (NifV-NifW-), indicating a possible role of NifW either in homocitrate transport or in the incorporation of this compound into the iron-molybdenum cofactor of nitrogenase. PMID:8492805

  10. System-reservoir theory with anharmonic baths: a perturbative approach

    NASA Astrophysics Data System (ADS)

    Bhadra, Chitrak; Banerjee, Dhruba

    2016-04-01

    In this paper we develop the formalism of a general system coupled to a reservoir (the words ‘bath’ and ‘reservoir’ will be used interchangeably) consisting of nonlinear oscillators, based on perturbation theory at the classical level, by extending the standard Zwanzig approach of elimination of bath degrees of freedom order by order in perturbation. We observe that the fluctuation dissipation relation (FDR) of the second kind in its standard form for harmonic baths gets modified due to the nonlinearity and this is manifested through higher powers of {{k}\\text{B}}T in the expression for two-time noise correlation. On the flip side, this very modification allows us to define a dressed (renormalized) system-bath coupling that depends on the temperature and the nonlinear parameters of the bath in such a way that the structure of the FDR (of the second kind) is maintained. As an aside, we also observe that the first moment of the noise arising from a nonlinear bath can be non-zero, even in the absence of any external drive, if the reservoir potential is asymmetric with respect to one of its minima, about which one builds up the perturbation theory.

  11. Environmental and behavioral conditions of bathing among elderly Japanese.

    PubMed

    Takasaki, Yuji; Ohnaka, Tadakatsu; Tochihara, Yutaka; Nagai, Yumiko; Ito, Hiromitsu; Yoshitake, Shiro

    2007-03-01

    This study investigated the bathing conditions of elderly Japanese, and sought to find factors relating to regional differences in death rates from bathtub accidents. A questionnaire survey was carried out in 11 areas of Japan. Questionnaires including questions regarding the length of time since houses had been built, types of facilities, and subjects' indoor thermal sensations and behavior while bathing were distributed to detached houses in each area twice, once in summer and once in winter. Completed questionnaires were collected from approximately 160 elderly people over 65 years old. Information regarding thermal sensations of rooms in winter revealed that a prefabricated bath and insulating window glass eased the cold in the bathroom. Unexpectedly, more subjects in the southern region than in the northern region reported being cold or a little cold while bathing in winter. In the present study, thermal sensations and behaviors while bathing seemed to be more affected by facilities and the location of houses than by the sex and age of the subjects. PMID:17435371

  12. Mutations conferring resistance to quinol oxidation (Qz) inhibitors of the cyt bc1 complex of Rhodobacter capsulatus.

    PubMed Central

    Daldal, F; Tokito, M K; Davidson, E; Faham, M

    1989-01-01

    Several spontaneous mutants of the photosynthetic bacterium Rhodobacter capsulatus resistant to myxothiazol, stigmatellin and mucidin--inhibitors of the ubiquinol: cytochrome c oxidoreductase (cyt bc1 complex)--were isolated. They were grouped into eight different classes based on their genetic location, growth properties and inhibitor cross-resistance. The petABC (fbcFBC) cluster that encodes the structural genes for the Rieske FeS protein, cyt b and cyt c1 subunits of the cyt bc1 complex was cloned out of the representative isolates and the molecular basis of inhibitor-resistance was determined by DNA sequencing. These data indicated that while one group of mutations was located outside the petABC(fbcFBC) cluster, the remainder were single base pair changes in codons corresponding to phylogenetically conserved amino acid residues of cyt b. Of these substitutions, F144S conferred resistance to myxothiazol, T163A and V333A to stigmatellin, L106P and G152S to myxothiazol + mucidin and M140I and F144L to myxothiazol + stigmatellin. In addition, a mutation (aer126) which specifically impairs the quinol oxidase (Qz) activity of the cyt bc1 complex of a non-photosynthetic mutant (R126) was identified to be a glycine to an aspartic acid replacement at position 158 of cyt b. Six of these mutations were found between amino acid residues 140 and 163, in a region linking the putative third and fourth transmembrane helices of cyt b. The non-random clustering of several inhibitor-resistance mutations around the non-functional aer126 mutation suggests that this region may be involved in the formation of the Qz inhibitor binding/quinol oxidation domain(s) of the cyt bc1 complex. Of the two remaining mutations, the V333A replacement conferred resistance to stigmatellin exclusively and was located in another region toward the C terminus of cyt b. The L106P substitution, on the other hand, was situated in the transmembrane helix II that carries two conserved histidine residues

  13. Analysis methods for meso- and macroporous silicon etching baths

    PubMed Central

    2012-01-01

    Analysis methods for electrochemical etching baths consisting of various concentrations of hydrofluoric acid (HF) and an additional organic surface wetting agent are presented. These electrolytes are used for the formation of meso- and macroporous silicon. Monitoring the etching bath composition requires at least one method each for the determination of the HF concentration and the organic content of the bath. However, it is a precondition that the analysis equipment withstands the aggressive HF. Titration and a fluoride ion-selective electrode are used for the determination of the HF and a cuvette test method for the analysis of the organic content, respectively. The most suitable analysis method is identified depending on the components in the electrolyte with the focus on capability of resistance against the aggressive HF. PMID:22805742

  14. Displacement of an Electrically Charged Drop on a Vibrating Bath.

    PubMed

    Brandenbourger, M; Vandewalle, N; Dorbolo, S

    2016-01-29

    In this work, the manipulation of an electrically charged droplet bouncing on a vertically vibrated bath is investigated. When a horizontal, uniform, and static electric field is applied to it, a motion is induced. The droplet is accelerated when the droplet is small. On the other hand, large droplets appear to move with a constant speed that depends linearly on the applied electrical field. In the latter regime, high-speed imaging of one bounce reveals that the droplet experiences an acceleration due to the electrical force during the flight and decelerates to 0 when interacting with the surface of the bath. Thus, the droplet moves with a constant average speed on a large time scale. We propose a criterion based on the force necessary to move a charged droplet at the surface of the bath to discriminate between constant speed and accelerated droplet regimes. PMID:26871337

  15. [Assessment of a hospital bath chair: an ergonomic approach].

    PubMed

    Comélio, Maria Estevam; Alexandre, Neusa Maria Costa

    2005-01-01

    A high rate of musculoskeletal disorders in members of the nursing team is observed in the literature. The use of special devices such as the bath chair has reduced the risk of back injuries in these workers and has also provided the patient with greater safety as well as comfort. The aim of the present study was to assess the ergonomic characteristics of a bath chair utilized in a hospital Separate questionnaires were applied for the nurses and the patients. The bath chair accessories presented several ergonomic problems. The perceived physical exertion according to the Borg-CR10 Scale indicated that a high level of exertion was required to manipulate the equipment (8.33). PMID:16514945

  16. Effects of Water Temperature during Foot Bath in Young Females.

    PubMed

    Nishimura, Masahiro; Tatsuya Saito, Tatsuya Saito; Kato, Toshiaki; Onodera, Sho

    2013-09-01

    We examined the effects of environmental and water temperatures of foot baths on pulse rate, blood pressure, mean skin temperature, salivary amylase (SA) activity, relaxation level and thermal sensation during winter. Five females participated in the study. The subjects rested in a chair for 20 min and the above-noted physiological reactions during the last 5 min of the resting period were recorded as baseline (BASE) values. Next, the subjects received a 15-min foot bath in water at 40 °C (WT40) or 45 °C (WT45), with a 15-min recovery period. Although SA is thought to be an indicator of stress via the sympathetic nervous system, we did not find a correlation between SA activity and relaxation state. We considered the possible effect of seasonal variation on the physiological reaction to foot bathing. PMID:24174706

  17. Production of aqueous spherical gold nanoparticles using conventional ultrasonic bath

    PubMed Central

    2012-01-01

    A conventional ultrasonic bath was used to examine the feasibility of forming aqueous spherical gold nanoparticles (GNPs) under atmospheric conditions. The effects of ultrasonic energy on the size and morphology of GNPs were also investigated. Highly monodispersed spherical GNPs were successfully synthesised by sodium citrate reduction in a conventional ultrasonic bath, without an additional heater or magnetic stirrer, as evidenced by ultraviolet–visible spectra and transmission electron microscopy. Ultrasonic energy was shown to be a key parameter for producing spherical GNPs of tunable sizes (20 to 50 nm). A proposed scheme for understanding the role of ultrasonic energy in the formation and growth of GNPs was discussed. The simple single-step method using just a conventional ultrasonic bath as demonstrated in this study offers new opportunities in the production of aqueous suspensions of monodispersed spherical GNPs. PMID:22839598

  18. [A sudden death case in the bath with peculiar scald].

    PubMed

    Hosoe, H; Tanegashima, A; Yamamoto, H; Nishiyama, N; Ogura, T; Hirasawa, T; Fukunaga, T

    1999-06-01

    A 60-year-old man was found dead in the bathtub of his house on the 2nd of February. Erythema with a clear border was observed on the right side of the face, trunk and around the knees, though the left side of the face, the hips and the feet were normal. The water heater was set outside of the bath, and the heated water flowed from the heater to the bath through the upper pipe. In order to determine the mechanism of the peculiar scalding, changes in the water temperature in the bathtub were measured under the same conditions. An hour after turning on the water heater, the surface water temperature of the bath was 73 degrees C. Zonal gradation of the temperature was observed. At the level of the opening for the lower intake pipe, it was 50 degrees C. The border of the scalding was consistent with the water layer above 50 degrees C. PMID:10536442

  19. Reduced quantum dynamics with arbitrary bath spectral densities: Hierarchical equations of motion based on several different bath decomposition schemes

    SciTech Connect

    Liu, Hao; Zhu, Lili; Bai, Shuming; Shi, Qiang

    2014-04-07

    We investigated applications of the hierarchical equation of motion (HEOM) method to perform high order perturbation calculations of reduced quantum dynamics for a harmonic bath with arbitrary spectral densities. Three different schemes are used to decompose the bath spectral density into analytical forms that are suitable to the HEOM treatment: (1) The multiple Lorentzian mode model that can be obtained by numerically fitting the model spectral density. (2) The combined Debye and oscillatory Debye modes model that can be constructed by fitting the corresponding classical bath correlation function. (3) A new method that uses undamped harmonic oscillator modes explicitly in the HEOM formalism. Methods to extract system-bath correlations were investigated for the above bath decomposition schemes. We also show that HEOM in the undamped harmonic oscillator modes can give detailed information on the partial Wigner transform of the total density operator. Theoretical analysis and numerical simulations of the spin-Boson dynamics and the absorption line shape of molecular dimers show that the HEOM formalism for high order perturbations can serve as an important tool in studying the quantum dissipative dynamics in the intermediate coupling regime.

  20. Pediatric sink-bathing: a risk for scald burns.

    PubMed

    Baggott, Kaitlin; Rabbitts, Angela; Leahy, Nicole E; Bourke, Patrick; Yurt, Roger W

    2013-01-01

    Our burn center previously reported a significant incidence of scald burns from tap water among patients treated at the center. However, mechanism of these scalds was not investigated in detail. A recent series of pediatric patients who sustained scalds while bathing in the sink was noted. To evaluate the extent of these injuries and create an effective prevention program for this population, a retrospective study of bathing-related sink burns among pediatric patients was performed. Patients between the ages of 0 and 5.0 years who sustained scald burns while being bathed in the sink were included in this study. Sex, race, age, burn size, length of stay, and surgical procedures were reviewed. During the study period of January 2003 through August 2008, 56 patients who were scalded in the sink were admitted, accounting for 54% of all bathing-related scalds. Among these, 56% were boys and 45% were Hispanic. Mean age was 0.8 ± 0.1 years. Burn size and hospital length of stay averaged 5 ± 0.7% and 11 ± 1 days, respectively. Of this group, 10.7% required skin grafting. The overwhelming majority (94% of patients) were discharged home. The remaining patients were discharged to inpatient rehabilitation, foster care, and others. Pediatric scald burns sustained while bathing in a sink continue to be prevalent at our burn center. Because of limited space and the child's proximity to faucet handles and water flow, sinks are an unsafe location to bathe a child. While such practice may be necessary for some families, comprehensive burn prevention education must address this hazard. PMID:23412329

  1. DNA sequence and genetic analysis of the Rhodobacter capsulatus nifENX gene region: homology between NifX and NifB suggests involvement of NifX in processing of the iron-molybdenum cofactor.

    PubMed

    Moreno-Vivian, C; Schmehl, M; Masepohl, B; Arnold, W; Klipp, W

    1989-04-01

    Rhodobacter capsulatus genes homologous to Klebsiella pneumoniae nifE, nifN and nifX were identified by DNA sequence analysis of a 4282 bp fragment of nif region A. Four open reading frames coding for a 51,188 (NifE), a 49,459 (NifN), a 17,459 (NifX) and a 17,472 (ORF4) dalton protein were detected. A typical NifA activated consensus promoter and two imperfect putative NifA binding sites were located in the 377 bp sequence in front of the nifE coding region. Comparison of the deduced amino acid sequences of R. capsulatus NifE and NifN revealed homologies not only to analogous gene products of other organisms but also to the alpha and beta subunits of the nitrogenase iron-molybdenum protein. In addition, the R. capsulatus nifE and nifN proteins shared considerable homology with each other. The map position of nifX downstream of nifEN corresponded in R. capsulatus and K. pneumoniae and the deduced molecular weights of both proteins were nearly identical. Nevertheless, R. capsulatus NifX was more related to the C-terminal end of NifY from K. pneumoniae than to NifX. A small domain of approximately 33 amino acid residues showing the highest degree of homology between NifY and NifX was also present in all nifB proteins analyzed so far. This homology indicated an evolutionary relationship of nifX, nifY and nifB and also suggested that NifX and NifY might play a role in maturation and/or stability of the iron-molybdenum cofactor. The open reading frame (ORF4) downstream of nifX in R. capsulatus is also present in Azotobacter vinelandii but not in K. pneumoniae.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2747620

  2. Hygiene for the newborn--to bath or to wash?

    PubMed Central

    Hylén, A. M.; Karlsson, E.; Svanberg, L.; Walder, M.

    1983-01-01

    Appropriate skin care of newborns is performed partly to prevent infection but also for aesthetic and cleansing purposes. Skin care should involve cleansing with a non-toxic, non-abrasive neutral material. This study compared the relative risks and benefits of washing versus bathing with regard to bacterial colonization rate, clinical infection rate, body temperature and crying. The results confirm our clinical impression that bathing and washing routines do not differ with regard to signs of infection or other clinical complications. However, the washing routine does increase the babies heat loss and make them less comfortable. PMID:6663065

  3. Existence of an independent phonon bath in a quantum device

    NASA Astrophysics Data System (ADS)

    Pascal, L. M. A.; Fay, A.; Winkelmann, C. B.; Courtois, H.

    2013-09-01

    At low temperatures, the thermal wavelength of acoustic phonons in a metallic thin film on a substrate can widely exceed the film thickness. It is thus generally believed that a mesoscopic device operating at low temperature does not carry an individual phonon population. In this work, we provide direct experimental evidence for the thermal decoupling of phonons in a mesoscopic quantum device from its substrate phonon heat bath at a sub-Kelvin temperature. A simple heat balance model assuming an independent phonon bath following the usual electron-phonon and Kapitza coupling laws can account for all experimental observations.

  4. Vitamin B12 regulates photosystem gene expression via the CrtJ antirepressor AerR in Rhodobacter capsulatus

    PubMed Central

    Cheng, Zhuo; Li, Keran; Hammad, Loubna A.; Karty, Jonathan A.; Bauer, Carl E.

    2014-01-01

    Summary The tetrapyrroles heme, bacteriochlorophyll and cobalamin (B12) exhibit a complex interrelationship regarding their synthesis. In this study, we demonstrate that AerR functions as an antirepressor of the tetrapyrrole regulator CrtJ. We show that purified AerR contains B12 that is bound to a conserved histidine (His145) in AerR. The interaction of AerR to CrtJ was further demonstrated in vitro by pull down experiments using AerR as bait and quantified using microscale thermophoresis. DNase I DNA footprint assays show that AerR containing B12 inhibits CrtJ binding to the bchC promoter. We further show that bchC expression is greatly repressed in a B12 auxotroph of Rhodobacter capsulatus and that B12 regulation of gene expression is mediated by AerR’s ability to function as an antirepressor of CrtJ. This study thus provides a mechanism for how the essential tetrapyrrole, cobalamin controls the synthesis of bacteriochlorophyll, an essential component of the photosystem. PMID:24329562

  5. Putative Hydrogen Bond to Tyrosine M208 in Photosynthetic Reaction Centers from Rhodobacter capsulatus Significantly Slows Primary Charge Separation

    PubMed Central

    2015-01-01

    Slow, ∼50 ps, P* → P+HA– electron transfer is observed in Rhodobacter capsulatus reaction centers (RCs) bearing the native Tyr residue at M208 and the single amino acid change of isoleucine at M204 to glutamic acid. The P* decay kinetics are unusually homogeneous (single exponential) at room temperature. Comparative solid-state NMR of [4′-13C]Tyr labeled wild-type and M204E RCs show that the chemical shift of Tyr M208 is significantly altered in the M204E mutant and in a manner consistent with formation of a hydrogen bond to the Tyr M208 hydroxyl group. Models based on RC crystal structure coordinates indicate that if such a hydrogen bond is formed between the Glu at M204 and the M208 Tyr hydroxyl group, the −OH would be oriented in a fashion expected (based on the calculations by Alden et al., J. Phys. Chem.1996, 100, 16761–16770) to destabilize P+BA– in free energy. Alteration of the environment of Tyr M208 and BA by Glu M204 via this putative hydrogen bond has a powerful influence on primary charge separation. PMID:24902471

  6. Markovian master equation for a classical particle coupled with arbitrary strength to a harmonic bath.

    PubMed

    Gelin, Maxim F

    2014-12-01

    We consider a classical point particle bilinearly coupled to a harmonic bath. Assuming that the evolution of the particle is monitored on a timescale which is longer than the characteristic bath correlation time, we derive the Markovian master equation for the probability density of the particle. The relaxation operator of this master equation is evaluated analytically, without invoking the perturbation theory and the approximation of weak system-bath coupling. When the bath correlation time tends to zero, the Fokker-Planck equation is recovered. For a finite bath correlation time, the relaxation operator contains contributions of all orders in the system-bath coupling. PMID:25481131

  7. Electron spin decoherence in nuclear spin baths and dynamical decoupling

    SciTech Connect

    Zhao, N.; Yang, W.; Ho, S. W.; Hu, J. L.; Wan, J. T. K.; Liu, R. B.

    2011-12-23

    We introduce the quantum theory of the electron spin decoherence in a nuclear spin bath and the dynamical decoupling approach for protecting the electron spin coherence. These theories are applied to various solid-state systems, such as radical spins in molecular crystals and NV centers in diamond.

  8. 21 CFR 890.5125 - Nonpowered sitz bath.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Nonpowered sitz bath. 890.5125 Section 890.5125 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES PHYSICAL MEDICINE DEVICES Physical Medicine Therapeutic Devices § 890.5125 Nonpowered...

  9. 21 CFR 890.5125 - Nonpowered sitz bath.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Nonpowered sitz bath. 890.5125 Section 890.5125 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES PHYSICAL MEDICINE DEVICES Physical Medicine Therapeutic Devices § 890.5125 Nonpowered...

  10. MASTER BATH SHOWING SINK WITH VANITY AND THE MEDICINE CABINET. ...

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

    MASTER BATH SHOWING SINK WITH VANITY AND THE MEDICINE CABINET. VIEW FACING WEST - Camp H.M. Smith and Navy Public Works Center Manana Title VII (Capehart) Housing, U-Shaped Two-Bedroom Single-Family Type 6, Birch Circle, Elm Drive, Elm Circle, and Date Drive, Pearl City, Honolulu County, HI

  11. 21 CFR 890.5125 - Nonpowered sitz bath.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Nonpowered sitz bath. 890.5125 Section 890.5125 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES PHYSICAL MEDICINE DEVICES Physical Medicine Therapeutic Devices § 890.5125 Nonpowered...

  12. 21 CFR 890.5125 - Nonpowered sitz bath.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Nonpowered sitz bath. 890.5125 Section 890.5125 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES PHYSICAL MEDICINE DEVICES Physical Medicine Therapeutic Devices § 890.5125 Nonpowered...

  13. Interior detail of unit "A" bath showing original medicine cabinet, ...

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

    Interior detail of unit "A" bath showing original medicine cabinet, ceramic soap dishes, ceramic towel rod, and triangular motif on ceramic features, facing south. - Albrook Air Force Station, Non-Commissioned Officers' Duplex, East side of Hall Street, Balboa, Former Panama Canal Zone, CZ

  14. Regenerate metal-plating baths to cut waste and save

    SciTech Connect

    1995-03-01

    During electrode-based metal plating of equipment components, the formation of an electrical field causes metal deposits to be thicker at edges and seams, and thinner on flat surfaces. And, electrode-based methods consume large amounts of energy. Electroless-nickel (EN) -- or autocatalytic -- plating systems were pioneered in the 1940s to solve these problems. EN plating produces a more uniform coating, irrespective of the complexity of the part, and it consumes less electricity, since to electric current is required during plating. Plating in an EN system results from a chemical reaction between nickel in the bath and the substrate of the equipment component. The downside of electroless plating, however, is the limited life of the nickel bath, and the large volume of metal waste produced by bath disposal. Ionsep Corp. (Wilmington, Del.) has developed an electrodialytic system that continuously reforms the EN plating baths, to give them longer life. Its patented system has been successfully laboratory tested in a 1-ft{sup 2} cell, and the firm recently won a $250,000 grant from the US Dept. of Energy (Washington, D.C.) and Environmental Protection Agency (Washington, D.C.), to design and engineer a commercial-scale version of the system.

  15. [Legionella pneumonia which occurred in a private whirlpool bath user].

    PubMed

    Ishikawa, Akira; Okada, Jun; Kondo, Hirobumi; Takayama, Youko; Sunagawa, Keisuke; Enari, Tadako; Ishii, Yoshikazu

    2004-10-01

    A 88 year old female with active rheumatoid arthritis treated by low dose of prednisolone and methotrexate was admitted to our hospital because of severe bilateral pulmonary infiltration and acute respiratory distress syndrome. On admission, she had consciousness disturbance and was intubated because of severe respiratory failure. We heard from her family of her habit she had taking a private whirlpool bath 2 or 3 times everyday. So, we suspected a Legionella pneumophila infection. We started intravenous erythromycin (EM) (1,500mg/day) and methylprednisolone pulse therapy (1,000mg x 3days) and full controlled mechanical ventilation supported with PEEP. Her respiratory failure was gradually improved and she was discharged on the 44 the hospital day. Legionella pneumophila (serogroup 6) was isolated in her sputum by B-CYE alpha culture. Legionella pneumophila (serogroup 6) was isolated in her private whirlpool bath too. Both samples revealed the same by genetic analysis with pulse field gel electrophoresis (PFGE). This is the first adult case of Legionella pneumophila pneumonia infected from a private whirlpool bath confirmed by genetic analysis. We should always suspect Legionella pneumonia as one of the severe community-acquired pneumonia, because Legionella pneumophila were frequently detected among various water sources including the private whirlpool bath. PMID:15560380

  16. 21 CFR 890.5125 - Nonpowered sitz bath.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Nonpowered sitz bath. 890.5125 Section 890.5125 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES PHYSICAL MEDICINE DEVICES Physical Medicine Therapeutic Devices § 890.5125 Nonpowered...

  17. A to Stirred-Liquid-Bath-Based Blackbody Source

    NASA Astrophysics Data System (ADS)

    Wang, J.; Yuan, Z.; Hao, X.; Wang, T.; Duan, Y.

    2015-08-01

    At the national facility for blackbody source radiance temperature calibration of the National Institute of Metrology, China, a stirred liquid bath blackbody was developed for use as a radiance temperature reference source, which has a temperature range from to . This blackbody source consists of a stirred liquid bath, a blackbody cavity, a standard capsule platinum resistance thermometer, and a dry-air purging system. The cavity is cylindrical with grooves on the inner wall. The cavity is 80 mm in diameter, with a depth of 520 mm, and is immersed in a bath filled with a water-ethylene glycol mixture. The average normal emissivity of the cavity is calculated to be better than 0.9999 with V grooves and when painted with Nextel 811-21 coating. The temperature stability of the blackbody source is over a period of 20 min, and the temperature uniformity of the cavity bottom is . The standard uncertainty of the radiance temperature of the stirred liquid bath blackbody source is estimated to be.

  18. VIEW OF INTEGRITY TESTING EQUIPMENT UTILIZING CRYOGENIC BATHS IN BUILDING ...

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

    VIEW OF INTEGRITY TESTING EQUIPMENT UTILIZING CRYOGENIC BATHS IN BUILDING 991. (6/7/68) - Rocky Flats Plant, Final Assembly & Shipping, Eastern portion of plant site, south of Spruce Avenue, east of Tenth Street & north of Central Avenue, Golden, Jefferson County, CO

  19. 75 FR 51177 - Safety Standard for Infant Bath Seats; Correction

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-19

    ..., 2010 (75 FR 31691). The document established a standard for infant bath seats by incorporating by... published in the Federal Register of June 4, 2010 (75 FR 31691) a final rule establishing a standard for... final rule, this phrase is redundant, and the final rule, therefore eliminates it.'' 75 FR...

  20. 9. VIEW OF MOLTEN SALT BATH EQUIPMENT AND ROLLER PRESSES ...

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

    9. VIEW OF MOLTEN SALT BATH EQUIPMENT AND ROLLER PRESSES BEING INSTALLED ON THE WEST SIDE (SIDE B) OF BUILDING 883. SIDE B OF BUILDING 883 WAS USED TO PROCESS ENRICHED URANIUM FROM 1957-66. (1/23/57) - Rocky Flats Plant, Uranium Rolling & Forming Operations, Southeast section of plant, southeast quadrant of intersection of Central Avenue & Eighth Street, Golden, Jefferson County, CO

  1. 13. VIEW OF THE MOLTEN SALT BATHS USED TO UNIFORMLY ...

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

    13. VIEW OF THE MOLTEN SALT BATHS USED TO UNIFORMLY AND QUICKLY HEAT METALS PRIOR TO WORKING (ROLLING). (9/16/85) - Rocky Flats Plant, Uranium Rolling & Forming Operations, Southeast section of plant, southeast quadrant of intersection of Central Avenue & Eighth Street, Golden, Jefferson County, CO

  2. Finite Systems in a Heat Bath: Spectrum Perturbations and Thermodynamics.

    PubMed

    de Miguel, Rodrigo; Rubi, J Miguel

    2016-09-01

    When a finite system is at equilibrium with a heat bath, the equilibrium temperature is dictated by the heat bath and not by the intrinsic thermostatistics of the finite system. If not sufficiently large, it may be necessary for the finite system to change its thermostatistics in order to be at equilibrium with the heat bath. We account for this process by invoking Landsberg's notion of temperature-dependent energy levels. We establish that the mismatch between the intrinsic temperature of the excited finite system and that of the heat bath drives a spectrum perturbation which enables thermal equilibrium. We show that the temperature-induced spectrum perturbation is equivalent to Hill's purely thermodynamic subdivision potential. The difference between intrinsic and equilibrium temperature provides us with a measure for how large a system can be before it no longer needs to be regarded as small. The theoretical framework proposed in this paper identifies the role of temperature in a bottom-up thermostatistical description of finite systems. PMID:27494276

  3. Bath osmolality: effect on water permeability of epithelial tissue.

    PubMed

    Lau, Y T; Parsons, R H; Feeney, G A; Walker, K L

    1982-03-01

    When hyperosmotic gradients from 100 to 500 mosM are used to produce a water flux, the water permeability of live and potassium cyanide (KCN)-poisoned frog skin decreases with increasing osmotic gradients. In addition, as the total bath osmolality (corium + epithelial) increases there is a reduction in tissue water. Examination of the tissue cellular and extracellular compartments shows that cell shrinkage caused by the increasing hyperosmolality of the bathing medium correlates with the decrease in osmotic permeability. When the bath osmolality is held constant and cell volume decreases, there is a decrease in the water permeability. High potassium in the external bathing medium causes cell swelling that is associated with an increase in water permeability. These data support the hypothesis that a number of conditions known to affect the water permeability of frog skin do so partly or wholly as a result of a change in the cell volume, which either directly or indirectly alters the osmotic permeability of a rate limiting barrier, possibly the cell membrane. PMID:6801996

  4. MASTER BATH SHOWING THE LINEN CLOSET WITH BUILTIN SHELVES. NOTE ...

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

    MASTER BATH SHOWING THE LINEN CLOSET WITH BUILT-IN SHELVES. NOTE THE WINDOWS IN THE UPPER PORTION OF THE EXTERIOR WALL. VIEW FACING NORTHWEST - Camp H.M. Smith and Navy Public Works Center Manana Title VII (Capehart) Housing, U-Shaped Two-Bedroom Duplex Type 1, Acacia Road, Birch Circle, and Cedar Drive, Pearl City, Honolulu County, HI

  5. 19. VIEW OF THE PLATING BATHS AND CONTROL PANELS. GOLD ...

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

    19. VIEW OF THE PLATING BATHS AND CONTROL PANELS. GOLD AND SILVER WERE AMONG THE MATERIALS PLATED ONTO PARTS MADE OF COPPER, STAINLESS STEEL AND STEEL. (11/15/89) - Rocky Flats Plant, Non-Nuclear Production Facility, South of Cottonwood Avenue, west of Seventh Avenue & east of Building 460, Golden, Jefferson County, CO

  6. Bath County Computer Attitude Scale: A Reliability and Validity Scale.

    ERIC Educational Resources Information Center

    Moroz, Pauline A.; Nash, John B.

    The Bath County Computer Attitude Scale (BCCAS) has received limited attention concerning its reliability and validity with a U.S. adult population. As developed by G. G. Bear, H. C. Richards, and P. Lancaster in 1987, the instrument assessed attitudes toward computers in areas of computer use, computer-aided instruction, programming and technical…

  7. Zinc recovery and waste sludge minimization from chromium passivation baths.

    PubMed

    Diban, Nazely; Mediavilla, Rosa; Urtiaga, Ane; Ortiz, Inmaculada

    2011-08-30

    This work reports the feasibility of applying emulsion pertraction technology (EPT) aiming at zinc recovery and waste minimization in the zinc electroplating processes that include Cr (III) passivation. The assessment consists of firstly the lifetime extension of the passivation baths by selective removal of the tramp ions zinc and iron, and secondly, the recovery of zinc for further reuse. Spent passivation baths from a local industry were tested, being the major metallic content: Cr(3+) 9000mg L(-1), Zn(2+) 12,000mg L(-1), Fe(3+) 100mg L(-1). Working in a Liqui-Cel hollow fiber membrane contactor and using the extractant bis(2,4,4-trimethylpentyl) phosphinic acid, reduction of zinc and iron concentrations below 60mg L(-1) and 2mg L(-1), respectively were obtained, while trivalent chromium, the active metal that generates the passivation layer, was retained in the baths. Zinc was selectively transferred to an acidic stripping phase that in the experimental time reached a concentration of 157,000mg L(-1). Zinc recovery by electrowinning from the acidic stripping phase without any pretreatment of the electrolyte solution provided a purity of 98.5%, matching the lower commercial zinc grade. As a result of the extension of the life time of the passivation bath, significant environmental advantages are derived such as minimization of the volume of hazardous wastes and savings in the consumption of raw materials. PMID:21704452

  8. BATH 1 SHOWING THE SHOWER ENCLOSURE AND FLUSH DOOR OF ...

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

    BATH 1 SHOWING THE SHOWER ENCLOSURE AND FLUSH DOOR OF LINEN CLOSET. VIEW FACING SOUTH - Camp H.M. Smith and Navy Public Works Center Manana Title VII (Capehart) Housing, U-Shaped Three-Bedroom Duplex Type 4, Acacia Road, Birch Circle, Cedar Drive and Elm Drive, Pearl City, Honolulu County, HI

  9. MASTER BATH SHOWING THE LINEN CLOSET WITH BUILT IN SHELVES ...

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

    MASTER BATH SHOWING THE LINEN CLOSET WITH BUILT IN SHELVES NEXT TO THE SHOWER ENCLOSURE. VIEW FACING NORTHEAST - Camp H.M. Smith and Navy Public Works Center Manana Title VII (Capehart) Housing, Three-Bedroom Single-Family Types 8 and 11, Birch Circle, Elm Drive, Elm Circle, and Date Drive, Pearl City, Honolulu County, HI

  10. MASTER BATH. NOTE THE LINEN CLOSET DOOR TO THE RIGHT ...

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

    MASTER BATH. NOTE THE LINEN CLOSET DOOR TO THE RIGHT OF THE SHOWER ENCLOSURE. VIEW FACING NORTHEAST - Camp H.M. Smith and Navy Public Works Center Manana Title VII (Capehart) Housing, Three-Bedroom Single-Family Type 7, Birch Circle, Elm Drive, Elm Circle, and Date Drive, Pearl City, Honolulu County, HI

  11. BATH 1 SHOWING THE LINEN CLOSET DOOR. VIEW FACING SOUTHWEST ...

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

    BATH 1 SHOWING THE LINEN CLOSET DOOR. VIEW FACING SOUTHWEST - Camp H.M. Smith and Navy Public Works Center Manana Title VII (Capehart) Housing, M-Shaped Four-Bedroom Duplex Type 5, Birch Circle, Cedar Drive, Pearl City, Honolulu County, HI

  12. Effects of tub bathing procedures on preterm infants' behavior.

    PubMed

    Liaw, Jen-Jiuan; Yang, Luke; Yuh, Yeong-Seng; Yin, Ti

    2006-12-01

    Although medical advances have increased the survival rate of preterm infants, morbidity in terms of neurodevelopmental impairment has not decreased for this population. This results in caregivers having to reconsider how neonatal intensive care unit (NICU) caregiving impacts on preterm infants. The purpose of this study was to examine the effects of different phases of a routine tub bath on preterm infants' distress and state behavior in the NICU. The study used an exploratory repeated measures design that focused on preterm infants' distress and state behavior, and evaluated the effects of three phases of a routine tub bath, which were performed according to standard unit practice in the NICU. Thirteen nurses repeatedly bathed 12 infants on different days, and 64 baths were videotaped for the purpose of assessing the variety of distress behavior. The procedures of one bath could be categorized into three phases designated to Phases I, II, and III. The variables were measured by a preterm infant behavioral coding scheme developed for this research. The inter-rater reliability of the instrument ranged from .82 to .99. Mixed effects analysis of variance was used to analyze the differences among the bath phases in the occurrences of distress and state behavior. The results showed significant statistical difference among most distress behaviors during the three phases (e.g. "startle, jerk, tremor" F ratio = 25.62, p < .001; "finger splay, grasping, fisting" F ratio = 49.99, p < .001; "grimace" F ratio = 36.55, p < .001; "fussing or crying" F ratio = 25.27, p < .001), with the exception of "extension, arching and squirming". In particular, the occurrence of distress and state behavior increased significantly in phase II. Routine tub bathing not only disrupts preterm infants' sleep but also causes an increase in distress behavior. Preterm infants' stress increases with the intrusiveness of nursing procedures. NICU caregivers should consider the effects of routine

  13. Bath Stone - a Possible Global Heritage Stone from England

    NASA Astrophysics Data System (ADS)

    Marker, Brian

    2014-05-01

    The Middle Jurassic strata of England have several horizons of oolitic and bioclastic limestones that provide high quality dimension stone. One of the most important is found in and near the City of Bath. The Great Oolite Group (Upper Bathonian) contains the Combe Down and Bath Oolites, consisting of current bedded oolites and shelly oolites, that have been used extensively as freestones for construction nearby, for prestigious buildings through much of southern England and more widely. The stone has been used to some extent since Roman times when the city, then known as Aquae Sulis, was an important hot spa. The stone was used to a limited extent through medieval times but from the early 18th century onwards was exploited on a large scale through surface quarrying and underground mining. The City was extensively redeveloped in the 18th to early 19th century, mostly using Bath Stone, when the spas made it a fashionable resort. Buildings from that period include architectural "gems" such as the Royal Crescent and Pulteney Bridge, as well as the renovated Roman Baths. Many buildings were designed by some of the foremost British architects of the time. The consistent use of this stone gives the City an architectural integrity throughout. These features led to the designation of the City as a World Heritage Site. It is a requirement in current City planning policy documents that Bath Stone should be used for new building to preserve the appearance of the City. More widely the stone was used in major houses (e.g. Buckingham Palace and Apsley House in London; King's Pavilion in Brighton); civic buildings (e.g. Bristol Guildhall; Dartmouth Naval College in Devon); churches and cathedrals (e.g. Truro Cathedral in Cornwall); and engineered structures (e.g. the large Dundas Aqueduct on the Kennet and Avon Canal). More widely, Bath Stone has been used in Union Station in Washington DC; Toronto Bible College and the Town Hall at Cape Town, South Africa. Extraction declined in

  14. One for All or All for One: Heterogeneous Expression and Host Cell Lysis Are Key to Gene Transfer Agent Activity in Rhodobacter capsulatus

    PubMed Central

    Fogg, Paul C. M.; Westbye, Alexander B.; Beatty, J. Thomas

    2012-01-01

    The gene transfer agent (RcGTA) of Rhodobacter capsulatus is the model for a family of novel bacteriophage-related genetic elements that carry out lateral transfer of essentially random host DNA. Genuine and putative gene transfer agents have been discovered in diverse genera and are becoming recognized as potentially an important source of genetic exchange and microbial evolution in the oceans. Despite being discovered over 30 years ago, little is known about many essential aspects of RcGTA biology. Here, we validate the use of direct fluorescence reporter constructs, which express the red fluorescent protein mCherry in R. capsulatus. A construct containing the RcGTA promoter fused to mCherry was used to examine the single-cell expression profiles of wild type and RcGTA overproducer R. capsulatus populations, under different growth conditions and growth phases. The majority of RcGTA production clearly arises from a small, distinct sub-set of the population in the wild type strain and a larger sub-set in the overproducer. The most likely RcGTA release mechanism concomitant with this expression pattern is host cell lysis and we present direct evidence for the release of an intracellular enzyme accompanying RcGTA release. RcGTA ORF s is annotated as a ‘cell wall peptidase’ but we rule out a role in host lysis and propose an alternative function as a key contributor to RcGTA invasion of a target cell during infection. PMID:22916305

  15. l-Malyl-Coenzyme A/β-Methylmalyl-Coenzyme A Lyase Is Involved in Acetate Assimilation of the Isocitrate Lyase-Negative Bacterium Rhodobacter capsulatus

    PubMed Central

    Meister, Michael; Saum, Stephan; Alber, Birgit E.; Fuchs, Georg

    2005-01-01

    Cell extracts of Rhodobacter capsulatus grown on acetate contained an apparent malate synthase activity but lacked isocitrate lyase activity. Therefore, R. capsulatus cannot use the glyoxylate cycle for acetate assimilation, and a different pathway must exist. It is shown that the apparent malate synthase activity is due to the combination of a malyl-coenzyme A (CoA) lyase and a malyl-CoA-hydrolyzing enzyme. Malyl-CoA lyase activity was 20-fold up-regulated in acetate-grown cells versus glucose-grown cells. Malyl-CoA lyase was purified 250-fold with a recovery of 6%. The enzyme catalyzed not only the reversible condensation of glyoxylate and acetyl-CoA to l-malyl-CoA but also the reversible condensation of glyoxylate and propionyl-CoA to β-methylmalyl-CoA. Enzyme activity was stimulated by divalent ions with preference for Mn2+ and was inhibited by EDTA. The N-terminal amino acid sequence was determined, and a corresponding gene coding for a 34.2-kDa protein was identified and designated mcl1. The native molecular mass of the purified protein was 195 ± 20 kDa, indicating a homohexameric composition. A homologous mcl1 gene was found in the genomes of the isocitrate lyase-negative bacteria Rhodobacter sphaeroides and Rhodospirillum rubrum in similar genomic environments. For Streptomyces coelicolor and Methylobacterium extorquens, mcl1 homologs are located within gene clusters implicated in acetate metabolism. We therefore propose that l-malyl-CoA/β-methylmalyl-CoA lyase encoded by mcl1 is involved in acetate assimilation by R. capsulatus and possibly other glyoxylate cycle-negative bacteria. PMID:15687206

  16. Effects of bathing solution on tensile properties of the cornea.

    PubMed

    Hatami-Marbini, Hamed; Rahimi, Abdolrasol

    2014-03-01

    The cornea is a transparent tissue with the major functions of protecting the inner contents of the eye and refracting incoming light. The biomechanical properties of the cornea strongly depend on the microstructure and composition of the stromal layer, a hydrated bio-gel. The uniaxial strip testing is a convenient and well-accepted experimental technique for characterizing corneal material parameters. It is known that the water content of specimens in this method depends on the osmolality of the bathing solution. The present study was designed to investigate the effects of different bathing solutions on uniaxial tensile material properties of the cornea. The tensile behavior of bovine corneal samples was measured in six different bathing solutions, i.e., hypertonic solution (12% NaCl solution), common preserving isotonic solutions (e.g., phosphate buffer saline, ophthalmic balanced salt solution, and 0.9% NaCl solution), hypotonic solution (distilled water), and neutral solution (mineral oil). It was observed that the bathing solution had significant influence on the tensile behavior of the corneal samples. In particular, the specimens tested in bathing solutions causing less swelling had significantly stiffer tensile properties. Furthermore, a simple mathematical model based on Voigt composite material model was developed to represent the measured solution-dependent tensile properties. The present study suggests that extra attention should be paid to corneal thickness (hydration) in uniaxial tensile experiments. It also provides important data on tensile properties of the cornea; such information could significantly contribute to improving the accuracy of numerical predictions of corneal biomechanics. PMID:24333541

  17. A stochastic reorganizational bath model for electronic energy transfer

    SciTech Connect

    Fujita, Takatoshi E-mail: aspuru@chemistry.harvard.edu; Huh, Joonsuk; Aspuru-Guzik, Alán E-mail: aspuru@chemistry.harvard.edu

    2014-06-28

    Environmentally induced fluctuations of the optical gap play a crucial role in electronic energy transfer dynamics. One of the simplest approaches to incorporate such fluctuations in energy transfer dynamics is the well known Haken-Strobl-Reineker (HSR) model, in which the energy-gap fluctuation is approximated as white noise. Recently, several groups have employed molecular dynamics simulations and excited-state calculations in conjunction to account for excitation energies’ thermal fluctuations. On the other hand, since the original work of HSR, many groups have employed stochastic models to simulate the same transfer dynamics. Here, we discuss a rigorous connection between the stochastic and the atomistic bath models. If the phonon bath is treated classically, time evolution of the exciton-phonon system can be described by Ehrenfest dynamics. To establish the relationship between the stochastic and atomistic bath models, we employ a projection operator technique to derive the generalized Langevin equations for the energy-gap fluctuations. The stochastic bath model can be obtained as an approximation of the atomistic Ehrenfest equations via the generalized Langevin approach. Based on this connection, we propose a novel scheme to take account of reorganization effects within the framework of stochastic models. The proposed scheme provides a better description of the population dynamics especially in the regime of strong exciton-phonon coupling. Finally, we discuss the effect of the bath reorganization in the absorption and fluorescence spectra of ideal J-aggregates in terms of the Stokes shifts. We find a simple expression that relates the reorganization contribution to the Stokes shifts – the reorganization shift – to the ideal or non-ideal exciton delocalization in a J-aggregate. The reorganization shift can be described by three parameters: the monomer reorganization energy, the relaxation time of the optical gap, and the exciton delocalization length

  18. Psychoactive “bath salts”: not so soothing

    PubMed Central

    Baumann, Michael H.; Partilla, John S.; Lehner, Kurt R.

    2012-01-01

    Recently there has been a dramatic rise in the abuse of so-called “bath salts” products that are purchased as legal alternatives to illicit drugs like cocaine and 3,4-methylenedioxymethamphetamine (MDMA). Baths salts contain one or more synthetic derivatives of the naturally-occurring stimulant cathinone. Low doses of bath salts produce euphoria and increase alertness, but high doses or chronic use can cause serious adverse effects such as hallucinations, delirium, hyperthermia and tachycardia. Owing to the risks posed by bath salts, the governments of many countries have made certain cathinones illegal, namely: 4-methylmethcathinone (mephedrone), 3,4-methylenedioxymethcathinone (methylone) and 3,4-methylenedioxypyrovalerone (MDPV). Similar to other psychomotor stimulants, synthetic cathinones target plasma membrane transporters for dopamine (i.e., DAT), norepinephrine (i.e., NET) and serotonin (i.e, SERT). Mephedrone and methylone act as non-selective transporter substrates, thereby stimulating non-exocytotic release of dopamine, norepinephrine and serotonin. By contrast, MDPV acts as a potent blocker at DAT and NET, with little effect at SERT. Administration of mephedrone or methylone to rats increases extracellular concentrations of dopamine and serotonin in the brain, analogous to the effects of MDMA. Not surprisingly, synthetic cathinones elicit locomotor activation in rodents. Stimulation of dopamine transmission by synthetic cathinones predicts a high potential for addiction and may underlie clinical adverse effects. As popular synthetic cathinones are rendered illegal, new replacement cathinones are appearing in the marketplace. More research on the pharmacology and toxicology of abused cathinones is needed to inform public health policy and develop strategies for treating medical consequence of bath salts abuse. PMID:23178799

  19. Difluoromethane, a new and improved inhibitor of methanotrophy

    USGS Publications Warehouse

    Miller, L.G.; Sasson, C.; Oremland, R.S.

    1998-01-01

    Difluoromethane (HFC-32; DFM) is compared to acetylene and methyl fluoride as an inhibitor of methanotrophy in cultures and soils. DFM was found to be a reversible inhibitor of CH4 oxidation by Methylococcus capsulatus (Bath). Consumption of CH4 in soil was blocked by additions of low levels of DFM (0.03 kPa), and this inhibition was reversed by DFM removal. Although a small quantity of DFM was consumed during these incubations, its remaining concentration was sufficiently elevated to sustain inhibition. Methanogenesis in anaerobic soil slurries, including acetoclastic methanogenesis, was unaffected by levels of DFM which inhibit methanotrophy. Low levels of DFM (0.03 kPa) also inhibited nitrification and N2O production by soils. DFM is proposed as an improved inhibitor of CH4 oxidation over acetylene and/or methyl fluoride on the basis of its reversibility, its efficacy at low concentrations, its lack of inhibition of methanogenesis, and its low cost.

  20. Crystal structure of a membrane-bound metalloenzyme that catalyses the biological oxidation of methane

    SciTech Connect

    Lieberman, R.L.; Rosenzweig, A.C.

    2010-03-08

    Particulate methane monooxygenase (pMMO) is an integral membrane metalloenzyme that catalyses the conversion of methane to methanol. Knowledge of how pMMO performs this extremely challenging chemistry may have an impact on the use of methane as an alternative energy source by facilitating the development of new synthetic catalysts. We have determined the structure of pMMO from the methanotroph Methylococcus capsulatus (Bath) to a resolution of 2.8 {angstrom}. The enzyme is a trimer with an {alpha}{sub 3}{beta}{sub 3}{gamma}{sub 3} polypeptide arrangement. Two metal centres, modelled as mononuclear copper and dinuclear copper, are located in soluble regions of each pmoB subunit, which resembles cytochrome c oxidase subunit II. A third metal centre, occupied by zinc in the crystal, is located within the membrane. The structure provides new insight into the molecular details of biological methane oxidation.

  1. Methanotroph outer membrane preparation.

    PubMed

    Karlsen, Odd A; Berven, Frode S; Jensen, Harald B; Fjellbirkeland, Anne

    2011-01-01

    All presently known methanotrophs are gram-negative bacteria suggesting that they are surrounded by a two-layered membrane: an inner or cytoplasmic membrane and an outer membrane. In the methanotroph Methylococcus capsulatus (Bath), separation of the two membranes has allowed studies on protein and lipid composition of the outer membrane. Its outer membrane can be isolated from purified cell envelopes by selective solubilization of the inner membranes with the detergent Triton X-100. The proteins associated with the outer membrane can further be fractionated into integral and tightly associated proteins and peripheral loosely associated proteins. We present here protocols for this fractionation and show how the proteins associated with the outer leaflet of the outer membrane can be isolated and identified by whole-cell biotin surface labeling. PMID:21419921

  2. Cardiac infection and sepsis in 3 intravenous bath salts drug users.

    PubMed

    Belton, Patrick; Sharngoe, Tenzing; Maguire, F Michael; Polhemus, Mark

    2013-06-01

    The street drug "bath salts" are psychoactive mixtures of cathinone derivatives. We report 3 cases of disseminated Staphylococcus aureus infection with cardiac involvement (2 endocarditis and 1 pericarditis), secondary to intravenous bath salts use. PMID:23418275

  3. Dust-bathing behavior of laying hens in enriched colony housing systems and an aviary system.

    PubMed

    Louton, H; Bergmann, S; Reese, S; Erhard, M H; Rauch, E

    2016-07-01

    The dust-bathing behavior of Lohmann Selected Leghorn hens was compared in 4 enriched colony housing systems and in an aviary system. The enriched colony housing systems differed especially in the alignment and division of the functional areas dust bath, nest, and perches. Forty-eight-hour video recordings were performed at 3 time-points during the laying period, and focal animal sampling and behavior sampling methods were used to analyze the dust-bathing behavior. Focal animal data included the relative fractions of dust-bathing hens overall, of hens bathing in the dust-bath area, and of those bathing on the wire floor throughout the day. Behavior data included the number of dust-bathing bouts within a predefined time range, the duration of 1 bout, the number of and reasons for interruptions, and the number of and reasons for the termination of dust-bathing bouts. Results showed that the average duration of dust bathing varied between the 4 enriched colony housing systems compared with the aviary system. The duration of dust-bathing bouts was shorter than reported under natural conditions. A positive correlation between dust-bathing activity and size of the dust-bath area was observed. Frequently, dust baths were interrupted and terminated by disturbing influences such as pecking by other hens. This was especially observed in the enriched colony housing systems. In none of the observed systems, neither in the enriched colony housing nor in the aviary system, were all of the observed dust baths terminated "normally." Dust bathing behavior on the wire mesh rather than in the provided dust-bath area generally was observed at different frequencies in all enriched colony housing systems during all observation periods, but never in the aviary system. The size and design of the dust-bath area influenced the prevalence of dust-bathing behavior in that small and subdivided dust-bath areas reduced the number of dust-bathing bouts but increased the incidence of sham dust

  4. Dust-bathing behavior of laying hens in enriched colony housing systems and an aviary system

    PubMed Central

    Louton, H.; Bergmann, S.; Reese, S.; Erhard, M. H.; Rauch, E.

    2016-01-01

    The dust-bathing behavior of Lohmann Selected Leghorn hens was compared in 4 enriched colony housing systems and in an aviary system. The enriched colony housing systems differed especially in the alignment and division of the functional areas dust bath, nest, and perches. Forty-eight-hour video recordings were performed at 3 time-points during the laying period, and focal animal sampling and behavior sampling methods were used to analyze the dust-bathing behavior. Focal animal data included the relative fractions of dust-bathing hens overall, of hens bathing in the dust-bath area, and of those bathing on the wire floor throughout the day. Behavior data included the number of dust-bathing bouts within a predefined time range, the duration of 1 bout, the number of and reasons for interruptions, and the number of and reasons for the termination of dust-bathing bouts. Results showed that the average duration of dust bathing varied between the 4 enriched colony housing systems compared with the aviary system. The duration of dust-bathing bouts was shorter than reported under natural conditions. A positive correlation between dust-bathing activity and size of the dust-bath area was observed. Frequently, dust baths were interrupted and terminated by disturbing influences such as pecking by other hens. This was especially observed in the enriched colony housing systems. In none of the observed systems, neither in the enriched colony housing nor in the aviary system, were all of the observed dust baths terminated “normally.” Dust bathing behavior on the wire mesh rather than in the provided dust-bath area generally was observed at different frequencies in all enriched colony housing systems during all observation periods, but never in the aviary system. The size and design of the dust-bath area influenced the prevalence of dust-bathing behavior in that small and subdivided dust-bath areas reduced the number of dust-bathing bouts but increased the incidence of sham dust

  5. Crystal Structure and Characterization of Particulate Methane Monooxygenase from Methylocystis species Strain M

    PubMed Central

    Smith, Stephen M.; Rawat, Swati; Telser, Joshua; Hoffman, Brian M.; Stemmler, Timothy L.; Rosenzweig, Amy C.

    2012-01-01

    Particulate methane monooxygenase (pMMO) is an integral membrane metalloenzyme that oxidizes methane to methanol in methanotrophic bacteria. Previous biochemical and structural studies of pMMO have focused on preparations from Methylococcus capsulatus (Bath) and Methylosinus trichosporium OB3b. A pMMO from a third organism, Methylocystis species strain M, has been isolated and characterized. Both membrane-bound and solubilized Methylocystis sp. strain M pMMO contain ~2 copper ions per 100 kDa protomer and exhibit copper-dependent propylene epoxidation activity. Spectroscopic data indicate that Methylocystis sp. strain M pMMO contains a mixture of CuI and CuII, of which the latter exhibits two distinct type 2 CuII electron paramagnetic resonance (EPR) signals. Extended X-ray absorption fine structure (EXAFS) data are best fit with a mixture of Cu–O/N and Cu–Cu ligand environments with a Cu–Cu interaction at 2.52–2.64 Å. The crystal structure of Methylocystis sp. strain M pMMO was determined to 2.68 Å resolution and is the best quality pMMO structure obtained to date. It provides a revised model for the pmoA and pmoC subunits and has led to an improved model of M. capsulatus (Bath) pMMO. In these new structures, the intramembrane zinc/copper binding site has a different coordination environment from that in previous models. PMID:22013879

  6. Crystal structure and characterization of particulate methane monooxygenase from Methylocystis species strain M.

    PubMed

    Smith, Stephen M; Rawat, Swati; Telser, Joshua; Hoffman, Brian M; Stemmler, Timothy L; Rosenzweig, Amy C

    2011-11-29

    Particulate methane monooxygenase (pMMO) is an integral membrane metalloenzyme that oxidizes methane to methanol in methanotrophic bacteria. Previous biochemical and structural studies of pMMO have focused on preparations from Methylococcus capsulatus (Bath) and Methylosinus trichosporium OB3b. A pMMO from a third organism, Methylocystis species strain M, has been isolated and characterized. Both membrane-bound and solubilized Methylocystis sp. strain M pMMO contain ~2 copper ions per 100 kDa protomer and exhibit copper-dependent propylene epoxidation activity. Spectroscopic data indicate that Methylocystis sp. strain M pMMO contains a mixture of Cu(I) and Cu(II), of which the latter exhibits two distinct type 2 Cu(II) electron paramagnetic resonance (EPR) signals. Extended X-ray absorption fine structure (EXAFS) data are best fit with a mixture of Cu-O/N and Cu-Cu ligand environments with a Cu-Cu interaction at 2.52-2.64 Å. The crystal structure of Methylocystis sp. strain M pMMO was determined to 2.68 Å resolution and is the best quality pMMO structure obtained to date. It provides a revised model for the pmoA and pmoC subunits and has led to an improved model of M. capsulatus (Bath) pMMO. In these new structures, the intramembrane zinc/copper binding site has a different coordination environment from that in previous models. PMID:22013879

  7. Remediation of chromium(VI) by a methane-oxidizing bacterium.

    PubMed

    Al Hasin, Abubakr; Gurman, Stephen J; Murphy, Loretta M; Perry, Ashlee; Smith, Thomas J; Gardiner, Philip H E

    2010-01-01

    Methane-oxidizing bacteria are ubiquitous in the environment and are globally important in oxidizing the potent greenhouse gas methane. It is also well recognized that they have wide potential for bioremediation of organic and chlorinated organic pollutants, thanks to the wide substrate ranges of the methane monooxygenase enzymes that they produce. Here we have demonstrated that the well characterized model methanotroph Methylococcus capsulatus (Bath) is able to bioremediate chromium(VI) pollution over a wide range of concentrations (1.4-1000 mg L(-1) of Cr(6+)), thus extending the bioremediation potential of this major group of microorganisms to include an important heavy-metal pollutant. The chromium(VI) reduction reaction was dependent on the availability of reducing equivalents from the growth substrate methane and was partially inhibited by the metabolic poison sodium azide. X-ray spectroscopy showed that the cell-associated chromium was predominantly in the +3 oxidation state and associated with cell- or medium-derived moieties that were most likely phosphate groups. The genome sequence of Mc. capsulatus (Bath) suggests at least five candidate genes for the chromium(VI) reductase activity in this organism. PMID:20039753

  8. An assay for screening microbial cultures for chalkophore production.

    PubMed

    Yoon, Sukhwan; Kraemer, Stephan M; Dispirito, Alan A; Semrau, Jeremy D

    2010-04-01

    Methanotrophs, bacteria that utilize methane as their sole carbon and energy source, are known to have high requirements for copper. These bacteria have recently been found to synthesize a copper-chelating agent, or chalkophore, termed methanobactin. To aid in screening methanobactin production by methanotrophs, a plate assay developed from the chrome azurol S (CAS) assay for siderophore production, was modified. In the typical CAS assay, a colour change from blue to orange in iron-CAS plates is observed as iron (III) ion weakly bound to CAS is sequestered by siderophores with higher affinities. In our modified assay, iron (III) chloride of the original CAS solution was substituted with copper (II) chloride, and removal of copper from CAS caused a colour change from blue to yellow. Assay results indicated that of the four tested methanotrophs (Methylosinus trichosporium OB3b, Methylococcus capsulatus Bath, Methylomicrobium album BG8 and Methylocystis parvus OBBP), only M. trichosporium OB3b, M. capsulatus Bath and M. album BG8 produced chalkophores capable of competing with CAS for copper, while M. parvus OBBP did not or did not export sufficient concentrations of methanobactin for detection by this assay. It was also found using Fe-CAS plates that at least M. trichosporium OB3b and M. album BG8 produce siderophores. These results may be expanded for the detection of chalkophores in other microorganisms as well as for screening of putative mutants of chalkophore synthesis. PMID:23766081

  9. Crystal Structure and Characterization of Particulate Methane Monooxygenase from Methylocystis species Strain M

    SciTech Connect

    Smith, Stephen M.; Rawat, Swati; Telser, Joshua; Hoffman, Brian M.; Stemmler, Timothy L.; Rosenzweig, Amy C.

    2012-02-08

    Particulate methane monooxygenase (pMMO) is an integral membrane metalloenzyme that oxidizes methane to methanol in methanotrophic bacteria. Previous biochemical and structural studies of pMMO have focused on preparations from Methylococcus capsulatus (Bath) and Methylosinus trichosporium OB3b. A pMMO from a third organism, Methylocystis species strain M, has been isolated and characterized. Both membrane-bound and solubilized Methylocystis sp. strain M pMMO contain {approx}2 copper ions per 100 kDa protomer and exhibit copper-dependent propylene epoxidation activity. Spectroscopic data indicate that Methylocystis sp. strain M pMMO contains a mixture of Cu{sup I} and Cu{sup II}, of which the latter exhibits two distinct type 2 Cu{sup II} electron paramagnetic resonance (EPR) signals. Extended X-ray absorption fine structure (EXAFS) data are best fit with a mixture of Cu-O/N and Cu-Cu ligand environments with a Cu-Cu interaction at 2.52-2.64 {angstrom}. The crystal structure of Methylocystis sp. strain M pMMO was determined to 2.68 {angstrom} resolution and is the best quality pMMO structure obtained to date. It provides a revised model for the pmoA and pmoC subunits and has led to an improved model of M. capsulatus (Bath) pMMO. In these new structures, the intramembrane zinc/copper binding site has a different coordination environment from that in previous models.

  10. Biochemical characterization of MmoS, a sensor protein involved in copper-dependent regulation of soluble methane monooxygenase.

    PubMed

    Ukaegbu, Uchechi E; Henery, Shannon; Rosenzweig, Amy C

    2006-08-29

    Methane monooxygenase (MMO) enzymes catalyze the oxidation of methane to methanol in methanotrophic bacteria. Several strains of methanotrophs, including Methylococcus capsulatus (Bath), express a membrane-bound or particulate MMO (pMMO) at high copper-to-biomass ratios and a soluble MMO (sMMO) form when copper is limited. The mechanism of this "copper switch" is not understood. The mmoS gene, located downstream of the sMMO operon, encodes a sensor protein that is part of a two-component signaling system and has been proposed to play a role in the copper switch. MmoS from M. capsulatus (Bath) has been cloned, expressed, and purified. The purified protein is a tetramer of molecular mass 480 kDa. Optical spectra indicate that MmoS contains a flavin cofactor, identified as flavin adenine dinucleotide (FAD) by fluorescence spectroscopy and chromatographic analysis. The redox potential of the MmoS-bound FAD, which binds within the N-terminal PAS-PAC domains, is -290 +/- 2 mV at pH 8.0 and 25 degrees C. Despite extensive efforts, MmoS could not be loaded with Cu(I) or Cu(II), indicating that MmoS does not sense copper directly. These data suggest that MmoS functions as a redox sensor and provide new insight into the copper-mediated regulation of sMMO expression. PMID:16922494

  11. [Isolation of Mycobacterium avium complex from the "24-hour bath"].

    PubMed

    Saito, H; Murakami, K; Ishii, N; Kwon, H H

    2000-01-01

    The "24-HOUR BATH" is an apparatus which circulates the bath water, keeps it clean and warm, and makes it possible to take a bath at any time during the day or night. It consists of apparatus for cleaning (sponge or mesh filter and filter material), heating (ceramic heater), and sterilizing (UV lamp). Recently, three cases of skin disease due to M. avium infection in private homes, in which "24-HOUR BATH" water was suspected to be the source of infection, have been reported. We attempted to isolate M. avium complex from the water (32 specimens), sponge filter (29 specimens), and filter material (32 specimens) of the "24-HOUR BATH". One hundred-ml samples of bath water, and 50-ml samples of rinse from a sponge filter or filter material were centrifuged at 3000 rpm for 20 min. Sediment was suspended in distilled water and a smear was prepared, and then digested and decontaminated with 2% sodium hydroxide. The processed specimens were cultured on 2% Ogawa medium containing ofloxacin (1 microgram/ml) and ethambutol (2.5 micrograms/ml) for 8 weeks at 37 degrees C. Positive smears were 3 (9.4%), 25 (86.2%) and 25 (78.1%) specimens from the water, sponge and filter material, respectively. A few bacterial clumps were observed, especially in the sponge specimens. The number of positive culture was 5 (15.6%), 24 (82.8%) and 25 (78.1%) from the water, sponge and filter material, respectively. Among them the number of Runyon's Group III-positive cultures was 5 (100%), 22 (91.7%) and 20 (80%) in the water, sponge, and filter material specimens, respectively. In most cases, cultures were positive for both the sponge and filter material specimens. All of the Group III mycobacteria were smooth, grew at 28, 37, 42, and 45 degrees C, negative for niacin, nitrate reductase, semiquantitative catalase, urease and Tween80 hydrolysis, and positive for 68 degrees C catalase. All of the strains reacted with M. avium complex AccuProbe and M. avium AccuProbe, but none of the strains reacted

  12. INVESTIGATION INTO THE REJUVENATION OF SPENT ELECTROLESS NICKEL BATHS BY ELECTRODIALYSIS

    EPA Science Inventory

    Electroless nickel plating generates substantially more waste than other metal-finishing processes due to the inherent limited bath life and the need for regular bath disposal. Electrodialysis can be used to generate electroless nickel baths, but poor membrane permselectivity, l...

  13. 30 CFR 71.400 - Bathing facilities; change rooms; sanitary flush toilet facilities.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Bathing facilities; change rooms; sanitary... WORK AREAS OF UNDERGROUND COAL MINES Surface Bathing Facilities, Change Rooms, and Sanitary Flush Toilet Facilities at Surface Coal Mines § 71.400 Bathing facilities; change rooms; sanitary flush...

  14. 30 CFR 71.400 - Bathing facilities; change rooms; sanitary flush toilet facilities.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 30 Mineral Resources 1 2011-07-01 2011-07-01 false Bathing facilities; change rooms; sanitary... WORK AREAS OF UNDERGROUND COAL MINES Surface Bathing Facilities, Change Rooms, and Sanitary Flush Toilet Facilities at Surface Coal Mines § 71.400 Bathing facilities; change rooms; sanitary flush...

  15. The Medical Risks and Benefits of Sauna, Steam Bath, and Whirlpool Use.

    ERIC Educational Resources Information Center

    Duda, Marty

    1987-01-01

    Saunas, steam baths, and whirlpools--popular fixtures at health clubs--are safe means of relaxation if used properly. Ignoring the recommendations for moderate, commonsense enjoyment of these baths may expose users to health risks, including sudden death, arrhythmias, and skin infections. A guide to safe use of such baths is presented. (Author/CB)

  16. 40 CFR 420.80 - Applicability; description of the salt bath descaling subcategory.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... into publicly owned treatment works resulting from oxidizing and reducing salt bath descaling... 40 Protection of Environment 28 2010-07-01 2010-07-01 true Applicability; description of the salt... Salt Bath Descaling Subcategory § 420.80 Applicability; description of the salt bath...

  17. Main chain and side chain dynamics of a heme protein: 15N and 2H NMR relaxation studies of R. capsulatus ferrocytochrome c2.

    PubMed

    Flynn, P F; Bieber Urbauer, R J; Zhang, H; Lee, A L; Wand, A J

    2001-06-01

    A detailed characterization of the main chain and side chain dynamics in R. capsulatus ferrocytochrome c(2) derived from (2)H NMR relaxation of methyl group resonances is presented. (15)N relaxation measurements confirm earlier results indicating that R. capsulatus ferrocytochrome c(2) exhibits minor rotational anisotropy in solution. The current study is focused on the use of deuterium relaxation in side chain methyl groups, which has been shown to provide a detailed and accurate measure of internal dynamics. Results obtained indicate that the side chains of ferrocytochrome c(2) exhibit a wide range of motional amplitudes, but are more rigid than generally found in the interior of nonprosthetic group bearing globular proteins. This unusual rigidity is ascribed to the interactions of the protein with the large heme prosthetic group. This observation has significant implications for the potential of the heme-protein interface to modulate the redox properties of the protein and also points to the need for great precision in the design and engineering of heme proteins. PMID:11380250

  18. Analysis of the Rhodobacter capsulatus puc operon: the pucC gene plays a central role in the regulation of LHII (B800-850 complex) expression.

    PubMed Central

    Tichy, H V; Albien, K U; Gad'on, N; Drews, G

    1991-01-01

    Formation of the light harvesting complex B800-850 (LHII) of Rhodobacter capsulatus requires the expression of more than the three known genes specific for that complex (pucA, pucB and pucE) encoding the alpha, beta and gamma subunits of LHII, respectively. In this work evidence is presented that the product of the gene pucC, which is located downstream from pucA, is essential for high-level transcription of the pucBACDE operon and formation of LHII. Plasmids were constructed containing deletions in one or several puc genes and transferred to a pucC::Tn5 mutant in which the puc operon is not expressed. It was found that the LHII- phenotype of the mutant was due to the missing PucC protein and that all known puc genes are located in one operon. To dissect the pucC, pucD and pucE genes from pucB and pucA and independently regulate them, they were placed under control of the nifHDK promoter. Only under nitrogen-fixing growth conditions was the LHII- pucC::Tn5 mutant complemented by this construction. It is concluded that expression of pucC is essential for formation of the LHII complex in R.capsulatus. Analysis of the pucD and pucE genes led to the conclusion that the products of these genes stabilize the B800-850 complex. Images PMID:1717257

  19. Molecular dynamics with coupling to an external bath

    NASA Astrophysics Data System (ADS)

    Berendsen, H. J. C.; Postma, J. P. M.; van Gunsteren, W. F.; DiNola, A.; Haak, J. R.

    1984-10-01

    In molecular dynamics (MD) simulations the need often arises to maintain such parameters as temperature or pressure rather than energy and volume, or to impose gradients for studying transport properties in nonequilibrium MD. A method is described to realize coupling to an external bath with constant temperature or pressure with adjustable time constants for the coupling. The method is easily extendable to other variables and to gradients, and can be applied also to polyatomic molecules involving internal constraints. The influence of coupling time constants on dynamical variables is evaluated. A leap-frog algorithm is presented for the general case involving constraints with coupling to both a constant temperature and a constant pressure bath.

  20. Appendix B: Inventory of coniferous forests near Bath, New York

    NASA Technical Reports Server (NTRS)

    Stanturf, J. A.

    1980-01-01

    A zoom stereoscope was used to interpret aerial color photographs of the Finger Lakes region near Bath, New York, and areas of conifers were delineated on acetate sheets. Scale was determined for each photograph and units were converted to acres. Photographically enlarged positive transparencies of imagery from LANDSAT bands 5,6, and 7 for the southern portion of the study area were placed in a cold additive viewer and registered with each other to provide a composite image. A green filter was used on band 5, blue on band 6, and red on band 7. Conifers appeared at dark, reddish purple. Average was determined using a grid. Results show that the total confer stands within 50 miles of Bath is approximately 176,000 acres of which 60,000 acres are in Pennsylvania. The study was conducted to determine the feasibility of locating a particleboard manufacturing firm in the Southern Tier.

  1. Electron spin decoherence in silicon carbide nuclear spin bath

    NASA Astrophysics Data System (ADS)

    Yang, Li-Ping

    In this paper, we study the electron spin decoherence of single defects in silicon carbide (SiC) nuclear spin bath. We find that, although the natural abundance of 29Si (4.7 counter-intuitive result, is the suppression of heteronuclear-spin flip-flop process in finite magnetic field. Our results show that electron spin of defect centers in SiC are excellent candidates for solid state spin qubit in quantum information processing.

  2. Giving a Newborn a Bath in her Parents' Presence.

    PubMed

    Didry, Pascale; Didry, Emmanuelle

    2015-11-01

    Today, Sophie is working on the maternity ward. She is going to give Manon, David and Laura's first born, a bath. Manon was born on the day before. She weighs 3.350kg and is 49cm long. She has already got a lot of fuzzy brown hair. Both parents are looking forward to watching and learning how to care for their new baby. PMID:26548395

  3. Density matrix embedding in an antisymmetrized geminal power bath

    SciTech Connect

    Tsuchimochi, Takashi; Welborn, Matthew; Van Voorhis, Troy

    2015-07-14

    Density matrix embedding theory (DMET) has emerged as a powerful tool for performing wave function-in-wave function embedding for strongly correlated systems. In traditional DMET, an accurate calculation is performed on a small impurity embedded in a mean field bath. Here, we extend the original DMET equations to account for correlation in the bath via an antisymmetrized geminal power (AGP) wave function. The resulting formalism has a number of advantages. First, it allows one to properly treat the weak correlation limit of independent pairs, which DMET is unable to do with a mean-field bath. Second, it associates a size extensive correlation energy with a given density matrix (for the models tested), which AGP by itself is incapable of providing. Third, it provides a reasonable description of charge redistribution in strongly correlated but non-periodic systems. Thus, AGP-DMET appears to be a good starting point for describing electron correlation in molecules, which are aperiodic and possess both strong and weak electron correlation.

  4. Bath salt intoxication causing acute kidney injury requiring hemodialysis.

    PubMed

    Regunath, Hariharan; Ariyamuthu, Venkatesh Kumar; Dalal, Pranavkumar; Misra, Madhukar

    2012-10-01

    Traditional bath salts contain a combination of inorganic salts like Epsom salts, table salt, baking soda, sodium metaphosphate, and borax that have cleansing properties. Since 2010, there have been rising concerns about a new type of substance abuse in the name of "bath salts." They are beta-ketone amphetamine analogs and are derivates of cathinone, a naturally occurring amphetamine analog found in the "khat" plant (Catha edulis). Effects reported with intake included increased energy, empathy, openness, and increased libido. Serious adverse effects reported with intoxication included cardiac, psychiatric, and neurological signs and symptoms. Not much is known about the toxicology and metabolism of these compounds. They inhibit monoamine reuptake (dopamine, nor epinephrine, etc.) and act as central nervous system stimulants with high additive and abuse potential because of their clinical and biochemical similarities to effects from use of cocaine, amphetamine, and 3,4-methylenedioxy-N-methylamphetamine. Deaths associated with use of these compounds have also been reported. We report a case of acute kidney injury associated with the use of "bath salt" pills that improved with hemodialysis. PMID:23036036

  5. Controlling the quantum dynamics of a mesoscopic spin bath in diamond

    PubMed Central

    de Lange, Gijs; van der Sar, Toeno; Blok, Machiel; Wang, Zhi-Hui; Dobrovitski, Viatcheslav; Hanson, Ronald

    2012-01-01

    Understanding and mitigating decoherence is a key challenge for quantum science and technology. The main source of decoherence for solid-state spin systems is the uncontrolled spin bath environment. Here, we demonstrate quantum control of a mesoscopic spin bath in diamond at room temperature that is composed of electron spins of substitutional nitrogen impurities. The resulting spin bath dynamics are probed using a single nitrogen-vacancy (NV) centre electron spin as a magnetic field sensor. We exploit the spin bath control to dynamically suppress dephasing of the NV spin by the spin bath. Furthermore, by combining spin bath control with dynamical decoupling, we directly measure the coherence and temporal correlations of different groups of bath spins. These results uncover a new arena for fundamental studies on decoherence and enable novel avenues for spin-based magnetometry and quantum information processing. PMID:22536480

  6. Proteome Profiling of the Rhodobacter capsulatus Molybdenum Response Reveals a Role of IscN in Nitrogen Fixation by Fe-Nitrogenase

    PubMed Central

    Hoffmann, Marie-Christine; Wagner, Eva; Langklotz, Sina; Pfänder, Yvonne; Hött, Sina; Bandow, Julia E.

    2015-01-01

    ABSTRACT Rhodobacter capsulatus is capable of synthesizing two nitrogenases, a molybdenum-dependent nitrogenase and an alternative Mo-free iron-only nitrogenase, enabling this diazotroph to grow with molecular dinitrogen (N2) as the sole nitrogen source. Here, the Mo responses of the wild type and of a mutant lacking ModABC, the high-affinity molybdate transporter, were examined by proteome profiling, Western analysis, epitope tagging, and lacZ reporter fusions. Many Mo-controlled proteins identified in this study have documented or presumed roles in nitrogen fixation, demonstrating the relevance of Mo control in this highly ATP-demanding process. The levels of Mo-nitrogenase, NifHDK, and the Mo storage protein, Mop, increased with increasing Mo concentrations. In contrast, Fe-nitrogenase, AnfHDGK, and ModABC, the Mo transporter, were expressed only under Mo-limiting conditions. IscN was identified as a novel Mo-repressed protein. Mo control of Mop, AnfHDGK, and ModABC corresponded to transcriptional regulation of their genes by the Mo-responsive regulators MopA and MopB. Mo control of NifHDK and IscN appeared to be more complex, involving different posttranscriptional mechanisms. In line with the simultaneous control of IscN and Fe-nitrogenase by Mo, IscN was found to be important for Fe-nitrogenase-dependent diazotrophic growth. The possible role of IscN as an A-type carrier providing Fe-nitrogenase with Fe-S clusters is discussed. IMPORTANCE Biological nitrogen fixation is a central process in the global nitrogen cycle by which the abundant but chemically inert dinitrogen (N2) is reduced to ammonia (NH3), a bioavailable form of nitrogen. Nitrogen reduction is catalyzed by nitrogenases found in diazotrophic bacteria and archaea but not in eukaryotes. All diazotrophs synthesize molybdenum-dependent nitrogenases. In addition, some diazotrophs, including Rhodobacter capsulatus, possess catalytically less efficient alternative Mo-free nitrogenases, whose expression

  7. Decoherence of a single spin coupled to an interacting spin bath

    NASA Astrophysics Data System (ADS)

    Wu, Ning; Fröhling, Nina; Xing, Xi; Hackmann, Johannes; Nanduri, Arun; Anders, Frithjof B.; Rabitz, Herschel

    2016-01-01

    Decoherence of a central spin coupled to an interacting spin bath via inhomogeneous Heisenberg coupling is studied by two different approaches, namely an exact equations of motion (EOMs) method and a Chebyshev expansion technique (CET). By assuming a wheel topology of the bath spins with uniform nearest-neighbor X X -type intrabath coupling, we examine the central spin dynamics with the bath prepared in two different types of bath initial conditions. For fully polarized baths in strong magnetic fields, the polarization dynamics of the central spin exhibits a collapse-revival behavior in the intermediate-time regime. Under an antiferromagnetic bath initial condition, the two methods give excellently consistent central spin decoherence dynamics for finite-size baths of N ≤14 bath spins. The decoherence factor is found to drop off abruptly on a short time scale and approach a finite plateau value which depends on the intrabath coupling strength nonmonotonically. In the ultrastrong intrabath coupling regime, the plateau values show an oscillatory behavior depending on whether N /2 is even or odd. The observed results are interpreted qualitatively within the framework of the EOM and perturbation analysis. The effects of anisotropic spin-bath coupling and inhomogeneous intrabath bath couplings are briefly discussed. Possible experimental realization of the model in a modified quantum corral setup is suggested.

  8. Incidence of Symptoms and Accidents During Baths and Showers Among the Japanese General Public

    PubMed Central

    Hayasaka, Shinya; Shibata, Yosuke; Noda, Tatsuya; Goto, Yasuaki; Ojima, Toshiyuki

    2011-01-01

    Background Bathing is a deeply ingrained custom among Japanese; however, data on the incidence rate of symptoms and accidents during bathing have not yet been reported for the Japanese general public. Methods We conducted a population-based cross-sectional study of 617 Japanese adults who attended a specialized health checkup. Participants completed a self-administered questionnaire to assess weekly frequencies of bathtub bathing and showering and the frequency of symptoms/accidents (falling, loss of consciousness, and other) during these activities in the past year. We calculated the incidence rates of accidents per 10 000 baths/showers and 95% confidence intervals (CIs) and compared the clinical characteristics of participants who had symptoms/accidents with those who did not. Results The incidence rates of accidents per 10 000 bathtub baths and showers were 0.43 (95% CI: 0.22–0.84) and 0.24 (95% CI: 0.04–1.37). Although these rates are low, there were 740 000 bathtub bathing-related accidents in Japan, due to the fact that bathing is an almost-daily habit. There was no significant difference in clinical characteristics between groups Conclusions We collected basic information on the incidence of bathing-related accidents in Japan. Falls and loss of consciousness during bathing or showering can potentially lead to a serious accident, so the general public should be educated about the possibility of such accidents during bathing. PMID:21478641

  9. [History of hot spring bath treatment in China].

    PubMed

    Hao, Wanpeng; Wang, Xiaojun; Xiang, Yinghong; Gu Li, A Man; Li, Ming; Zhang, Xin

    2011-07-01

    As early as the 7th century B.C. (Western Zhou Dynasty), there is a recording as 'spring which contains sulfur could treat disease' on the Wentang Stele written by WANG Bao. Wenquan Fu written by ZHANG Heng in the Easten Han Dynasty also mentioned hot spring bath treatment. The distribution of hot springs in China has been summarized by LI Daoyuan in the Northern Wei Dynasty in his Shuijingzhu which recorded hot springs in 41 places and interpreted the definition of hot spring. Bencao Shiyi (by CHEN Cangqi, Tang Dynasty) discussed the formation of and indications for hot springs. HU Zai in the Song Dynasty pointed out distinguishing hot springs according to water quality in his book Yuyin Conghua. TANG Shenwei in the Song Dynasty noted in Jingshi Zhenglei Beiji Bencao that hot spring bath treatment should be combined with diet. Shiwu Bencao (Ming Dynasty) classified hot springs into sulfur springs, arsenicum springs, cinnabar springs, aluminite springs, etc. and pointed out their individual indications. Geologists did not start the work on distribution and water quality analysis of hot springs until the first half of the 20th century. There are 972 hot springs in Wenquan Jiyao (written by geologist ZHANG Hongzhao and published in 1956). In July 1982, the First National Geothermal Conference was held and it reported that there were more than 2600 hot springs in China. Since the second half of the 20th century, hot spring sanatoriums and rehabilitation centers have been established, which promoted the development of hot spring bath treatment. PMID:22169492

  10. Resummed memory kernels in generalized system-bath master equations

    SciTech Connect

    Mavros, Michael G.; Van Voorhis, Troy

    2014-08-07

    Generalized master equations provide a concise formalism for studying reduced population dynamics. Usually, these master equations require a perturbative expansion of the memory kernels governing the dynamics; in order to prevent divergences, these expansions must be resummed. Resummation techniques of perturbation series are ubiquitous in physics, but they have not been readily studied for the time-dependent memory kernels used in generalized master equations. In this paper, we present a comparison of different resummation techniques for such memory kernels up to fourth order. We study specifically the spin-boson Hamiltonian as a model system bath Hamiltonian, treating the diabatic coupling between the two states as a perturbation. A novel derivation of the fourth-order memory kernel for the spin-boson problem is presented; then, the second- and fourth-order kernels are evaluated numerically for a variety of spin-boson parameter regimes. We find that resumming the kernels through fourth order using a Padé approximant results in divergent populations in the strong electronic coupling regime due to a singularity introduced by the nature of the resummation, and thus recommend a non-divergent exponential resummation (the “Landau-Zener resummation” of previous work). The inclusion of fourth-order effects in a Landau-Zener-resummed kernel is shown to improve both the dephasing rate and the obedience of detailed balance over simpler prescriptions like the non-interacting blip approximation, showing a relatively quick convergence on the exact answer. The results suggest that including higher-order contributions to the memory kernel of a generalized master equation and performing an appropriate resummation can provide a numerically-exact solution to system-bath dynamics for a general spectral density, opening the way to a new class of methods for treating system-bath dynamics.

  11. Resummed memory kernels in generalized system-bath master equations

    NASA Astrophysics Data System (ADS)

    Mavros, Michael G.; Van Voorhis, Troy

    2014-08-01

    Generalized master equations provide a concise formalism for studying reduced population dynamics. Usually, these master equations require a perturbative expansion of the memory kernels governing the dynamics; in order to prevent divergences, these expansions must be resummed. Resummation techniques of perturbation series are ubiquitous in physics, but they have not been readily studied for the time-dependent memory kernels used in generalized master equations. In this paper, we present a comparison of different resummation techniques for such memory kernels up to fourth order. We study specifically the spin-boson Hamiltonian as a model system bath Hamiltonian, treating the diabatic coupling between the two states as a perturbation. A novel derivation of the fourth-order memory kernel for the spin-boson problem is presented; then, the second- and fourth-order kernels are evaluated numerically for a variety of spin-boson parameter regimes. We find that resumming the kernels through fourth order using a Padé approximant results in divergent populations in the strong electronic coupling regime due to a singularity introduced by the nature of the resummation, and thus recommend a non-divergent exponential resummation (the "Landau-Zener resummation" of previous work). The inclusion of fourth-order effects in a Landau-Zener-resummed kernel is shown to improve both the dephasing rate and the obedience of detailed balance over simpler prescriptions like the non-interacting blip approximation, showing a relatively quick convergence on the exact answer. The results suggest that including higher-order contributions to the memory kernel of a generalized master equation and performing an appropriate resummation can provide a numerically-exact solution to system-bath dynamics for a general spectral density, opening the way to a new class of methods for treating system-bath dynamics.

  12. Resummed memory kernels in generalized system-bath master equations.

    PubMed

    Mavros, Michael G; Van Voorhis, Troy

    2014-08-01

    Generalized master equations provide a concise formalism for studying reduced population dynamics. Usually, these master equations require a perturbative expansion of the memory kernels governing the dynamics; in order to prevent divergences, these expansions must be resummed. Resummation techniques of perturbation series are ubiquitous in physics, but they have not been readily studied for the time-dependent memory kernels used in generalized master equations. In this paper, we present a comparison of different resummation techniques for such memory kernels up to fourth order. We study specifically the spin-boson Hamiltonian as a model system bath Hamiltonian, treating the diabatic coupling between the two states as a perturbation. A novel derivation of the fourth-order memory kernel for the spin-boson problem is presented; then, the second- and fourth-order kernels are evaluated numerically for a variety of spin-boson parameter regimes. We find that resumming the kernels through fourth order using a Padé approximant results in divergent populations in the strong electronic coupling regime due to a singularity introduced by the nature of the resummation, and thus recommend a non-divergent exponential resummation (the "Landau-Zener resummation" of previous work). The inclusion of fourth-order effects in a Landau-Zener-resummed kernel is shown to improve both the dephasing rate and the obedience of detailed balance over simpler prescriptions like the non-interacting blip approximation, showing a relatively quick convergence on the exact answer. The results suggest that including higher-order contributions to the memory kernel of a generalized master equation and performing an appropriate resummation can provide a numerically-exact solution to system-bath dynamics for a general spectral density, opening the way to a new class of methods for treating system-bath dynamics. PMID:25106575

  13. Transport of thermal water from well to thermal baths

    NASA Astrophysics Data System (ADS)

    Montegrossi, Giordano; Vaselli, Orlando; Tassi, Franco; Nocentini, Matteo; Liccioli, Caterina; Nisi, Barbara

    2013-04-01

    The main problem in building a thermal bath is having a hot spring or a thermal well located in an appropriate position for customer access; since Roman age, thermal baths were distributed in the whole empire and often road and cities were built all around afterwards. Nowadays, the perspectives are changed and occasionally the thermal resource is required to be transported with a pipeline system from the main source to the spa. Nevertheless, the geothermal fluid may show problems of corrosion and scaling during transport. In the Ambra valley, central Italy, a geothermal well has recently been drilled and it discharges a Ca(Mg)-SO4, CO2-rich water at the temperature of 41 °C, that could be used for supplying a new spa in the surrounding areas of the well itself. The main problem is that the producing well is located in a forest tree ca. 4 km far away from the nearest structure suitable to host the thermal bath. In this study, we illustrate the pipeline design from the producing well to the spa, constraining the physical and geochemical parameters to reduce scaling and corrosion phenomena. The starting point is the thermal well that has a flow rate ranging from 22 up to 25 L/sec. The thermal fluid is heavily precipitating calcite (50-100 ton/month) due to the calcite-CO2 equilibrium in the reservoir, where a partial pressure of 11 bar of CO2 is present. One of the most vexing problems in investigating scaling processed during the fluid transport in the pipeline is that there is not a proper software package for multiphase fluid flow in pipes characterized by such a complex chemistry. As a consequence, we used a modified TOUGHREACT with Pitzer database, arranged to use Darcy-Weisbach equation, and applying "fictitious" material properties in order to give the proper y- z- velocity profile in comparison to the analytical solution for laminar fluid flow in pipes. This investigation gave as a result the lowest CO2 partial pressure to be kept in the pipeline (nearly 2

  14. Langevin description of gauged scalar fields in a thermal bath

    NASA Astrophysics Data System (ADS)

    Miyamoto, Yuhei; Motohashi, Hayato; Suyama, Teruaki; Yokoyama, Jun'ichi

    2014-04-01

    We study the dynamics of the oscillating gauged scalar field in a thermal bath. A Langevin-type equation of motion of the scalar field, which contains both dissipation and fluctuation terms, is derived by using the real-time finite-temperature effective action approach. The existence of the quantum fluctuation-dissipation relation between the nonlocal dissipation term and the Gaussian stochastic noise terms is verified. We find that the noise variables are anticorrelated at equal time. The dissipation rate for each mode is also studied, which turns out to depend on the wave number.

  15. Dissociation rate of bromine diatomics in an argon heat bath

    NASA Technical Reports Server (NTRS)

    Razner, R.; Hopkins, D.

    1973-01-01

    The evolution of a collection of 300 K bromine diatomics embedded in a heat bath of argon atoms at 1800 K was studied by computer, and a dissociation-rate constant for the reaction Br2 + BR + Ar yields Br + Ar was determined. Previously published probability distributions for energy and angular momentum transfers in classical three-dimensional Br2-Ar collisions were used in conjunction with a newly developed Monte Carlo scheme for this purpose. Results are compared with experimental shock-tube data and the predictions of several other theoretical models. A departure from equilibrium is obtained which is significantly greater than that predicted by any of these other theories.

  16. Detection of weak signals in memory thermal baths

    NASA Astrophysics Data System (ADS)

    Jiménez-Aquino, J. I.; Velasco, R. M.; Romero-Bastida, M.

    2014-11-01

    The nonlinear relaxation time and the statistics of the first passage time distribution in connection with the quasideterministic approach are used to detect weak signals in the decay process of the unstable state of a Brownian particle embedded in memory thermal baths. The study is performed in the overdamped approximation of a generalized Langevin equation characterized by an exponential decay in the friction memory kernel. A detection criterion for each time scale is studied: The first one is referred to as the receiver output, which is given as a function of the nonlinear relaxation time, and the second one is related to the statistics of the first passage time distribution.

  17. Verification of impact of morning showering and mist sauna bathing on human physiological functions and work efficiency during the day

    NASA Astrophysics Data System (ADS)

    Lee, Soomin; Fujimura, Hiroko; Shimomura, Yoshihiro; Katsuura, Tetsuo

    2015-09-01

    Recently, a growing number in Japan are switching to taking baths in the morning (morning bathing). However, the effects of the morning bathing on human physiological functions and work efficiency have not yet been revealed. Then, we hypothesized that the effect of morning bathing on physiological functions would be different from those of night bathing. In this study, we measured the physiological functions and work efficiency during the day following the morning bathing (7:10-7:20) including showering, mist sauna bathing, and no bathing as a control. Ten male healthy young adults participated in this study as the subjects. We evaluated the rectal temperature (Tre), skin temperature (Tsk), heart rate (HR), heart rate variability (HRV), blood pressure (BP), the relative power density of the alpha wave (α-wave ratio) of electroencephalogram, alpha attenuation coefficient (AAC), and the error rate of the task performance. As a result, we found that the HR after the mist sauna bathing was significantly lower than those after no bathing rest 3 (11:00). Furthermore, we verified that the α-wave ratio of the Pz after the mist sauna bathing was significantly lower than those after no bathing during the task 6 (15:00). On the other hand, the α-wave ratio of the Pz after the mist sauna bathing was significantly higher than those after showering during the rest 3 (11:00). Tsk after the mist sauna bathing was higher than those after the showering at 9:00 and 15:00. In addition, the error rate of the task performance after the mist sauna bathing was lower than those after no bathing and showering at 14:00. This study concludes that a morning mist sauna is safe and maintains both skin temperature compared to other bathing methods. Moreover, it is presumed that the morning mist sauna bathing improves work efficiency comparing other bathing methods during the task period of the day following the morning bathing.

  18. Experimental study of pharmacokinetics of external, whole-body bathing application of ivermectin.

    PubMed

    Miyajima, Atsushi; Komoda, Masayo; Akagi, Keita; Yuzawa, Kaoru; Yoshimasu, Takashi; Yamamoto, Yosuke; Hirota, Takashi

    2015-01-01

    As a novel method improving the safety of conventional oral ivermectin (IVM) for scabies treatment, we conceived an idea called the "whole-body bathing method". In this method, the patients would bathe themselves in a bathing fluid containing IVM at an effective concentration. To evaluate the feasibility of the method, we investigated the IVM concentration in the skin and plasma after bathing rats in a fluid containing 100 ng/mL of IVM. After the bathing, the concentration of IVM in the skin was more than 400 ng/g wet weight and was maintained until 8 h after the bathing. The concentration was clearly higher than that in patients taking IVM p.o. as previously reported; IVM was not detected in plasma in the present study. Thus, the method would be a preferable drug delivery system for the skin application of IVM compared with p.o. administration. PMID:25492083

  19. Vibrational spectroscopy of a harmonic oscillator system nonlinearly coupled to a heat bath

    NASA Astrophysics Data System (ADS)

    Kato, Tsuyoshi; Tanimura, Yoshitaka

    2002-10-01

    Vibrational relaxation of a harmonic oscillator nonlinearly coupled to a heat bath is investigated by the Gaussian-Markovian quantum Fokker-Planck equation approach. The system-bath interaction is assumed to be linear in the bath coordinate, but linear plus square in the system coordinate modeling the elastic and inelastic relaxation mechanisms. Interplay of the two relaxation processes induced by the linear-linear and square-linear interactions in Raman or infrared spectra is discussed for various system-bath couplings, temperatures, and correlation times for the bath fluctuations. The one-quantum coherence state created through the interaction with the pump laser pulse relaxes through different pathways in accordance with the mechanisms of the system-bath interactions. Relations between the present theory, Redfield theory, and stochastic theory are also discussed.

  20. Music-assisted bathing: making shower time easier for people with dementia.

    PubMed

    Ray, Kendra D; Fitzsimmons, Suzanne

    2014-02-01

    It is estimated that 90% of nursing home residents need assistance with bathing. The purpose of this article is to describe a music-assisted care technique that can be used by caregivers when bathing nursing home residents with dementia. Research suggests that music has many therapeutic benefits for people with dementia. Using music to soothe anxiety can be an effective intervention to assist with lessening of agitation during activities of daily living, especially bathing. This article will provide nursing and direct care staff tools to successfully conduct the music-assisted bathing protocol. Consideration for choosing appropriate music for bathing, the creation of individualized personalized playlists, and acknowledgement of desired outcomes are presented. Incorporating music-assisted bathing may address neuropsychiatric symptoms of dementia by lessening agitation and improving mood, which in turn can increase job satisfaction. PMID:24550123

  1. Dissipative Landau-Zener transitions of a qubit: Bath-specific and universal behavior

    SciTech Connect

    Saito, Keiji; Wubs, Martijn; Kohler, Sigmund; Haenggi, Peter; Kayanuma, Yosuke

    2007-06-01

    We study Landau-Zener transitions in a qubit coupled to a bath at zero temperature. A general formula that is applicable to models with a nondegenerate ground state is derived. We calculate exact transition probabilities for a qubit coupled to either a bosonic or a spin bath. The nature of the baths and the qubit-bath coupling is reflected in the transition probabilities. For diagonal coupling, when the bath causes energy fluctuations of the diabatic qubit states but no transitions between them, the transition probability coincides with the standard Landau-Zener probability of an isolated qubit. This result is universal as it does not depend on the specific type of bath. For pure off-diagonal coupling, by contrast, the tunneling probability is sensitive to the coupling strength. We discuss the relevance of our results for experiments on molecular nanomagnets, in circuit QED, and for the fast-pulse readout of superconducting phase qubits.

  2. Efficient heat-bath sampling in Fock space

    NASA Astrophysics Data System (ADS)

    Holmes, Adam; Changlani, Hitesh; Umrigar, Cyrus

    We introduce an algorithm for sampling many-body quantum states in Fock space. The algorithm efficiently samples states with probability approximately proportional to an arbitrary function of the second-quantized Hamiltonian matrix elements connected to the current state. We apply the new sampling algorithm to the recently-developed Semistochastic Full Configuration Interaction Quantum Monte Carlo method (S-FCIQMC), a semistochastic implementation of the power method for projecting out the ground state energy in a basis of Slater determinants. The heat-bath sampling requires modest additional computational time and memory compared to uniform sampling but results in newly-spawned weights that are approximately of the same magnitude, thereby greatly improving the efficiency of projection. A comparison in efficiency between uniform and approximate heat-bath sampling is performed on the all-electron nitrogen dimer at equilibrium in Dunning's cc-pVXZ basis sets with X ∈ D , T , Q , 5 , demonstrating a large gain in efficiency that increases with basis set size. This work was supported in part by grants NSF CHE-1112097, DOE DE-SC0006650, and NSF ACI-1534965.

  3. Achievable Polarization for Heat-Bath Algorithmic Cooling.

    PubMed

    Rodríguez-Briones, Nayeli Azucena; Laflamme, Raymond

    2016-04-29

    Pure quantum states play a central role in applications of quantum information, both as initial states for quantum algorithms and as resources for quantum error correction. Preparation of highly pure states that satisfy the threshold for quantum error correction remains a challenge, not only for ensemble implementations like NMR or ESR but also for other technologies. Heat-bath algorithmic cooling is a method to increase the purity of a set of qubits coupled to a bath. We investigated the achievable polarization by analyzing the limit when no more entropy can be extracted from the system. In particular, we give an analytic form for the maximum polarization achievable for the case when the initial state of the qubits is totally mixed, and the corresponding steady state of the whole system. It is, however, possible to reach higher polarization while starting with certain states; thus, our result provides an achievable bound. We also give the number of steps needed to get a specific required polarization. PMID:27176508

  4. Lunar Surface Systems Wet-Bath Design Evaluation

    NASA Technical Reports Server (NTRS)

    Thompson, Shelby; Szabo, Rich; Howard, Robert

    2010-01-01

    The goal of the current evaluation was to examine five different wet-bath architectural design concepts. The primary means of testing the concepts required participants to physically act-out a number of functional tasks (e.g., shaving, showering, changing clothes, maintenance) in order to give judgments on the affordance of the volume as based on the design concepts. Each of the concepts was designed in such a way that certain features were exploited - for example, a concept may have a large amount of internal stowage, but minimum amount of usable space to perform tasks. The results showed that the most preferred concept was one in which stowage and usable space were balanced. This concept allowed for a moderate amount of stowage with some suggested redesign, but would not preclude additional personal items such as clothing. This concept also allowed for a greater distance to be achieved between the toilet and the sink with minimum redesign, which was desirable. Therefore, the all-in-one (i.e., toilet, sink, and shower all occupying a single volume) wet-bath concept seemed to be a viable solution in which there is a minimal amount of overall volume available with certain lunar habitat configurations.

  5. Achievable Polarization for Heat-Bath Algorithmic Cooling

    NASA Astrophysics Data System (ADS)

    Rodríguez-Briones, Nayeli Azucena; Laflamme, Raymond

    2016-04-01

    Pure quantum states play a central role in applications of quantum information, both as initial states for quantum algorithms and as resources for quantum error correction. Preparation of highly pure states that satisfy the threshold for quantum error correction remains a challenge, not only for ensemble implementations like NMR or ESR but also for other technologies. Heat-bath algorithmic cooling is a method to increase the purity of a set of qubits coupled to a bath. We investigated the achievable polarization by analyzing the limit when no more entropy can be extracted from the system. In particular, we give an analytic form for the maximum polarization achievable for the case when the initial state of the qubits is totally mixed, and the corresponding steady state of the whole system. It is, however, possible to reach higher polarization while starting with certain states; thus, our result provides an achievable bound. We also give the number of steps needed to get a specific required polarization.

  6. Chemical bath deposition of II-VI compound thin films

    NASA Astrophysics Data System (ADS)

    Oladeji, Isaiah Olatunde

    II-VI compounds are direct bandgap semiconductors with great potentials in optoelectronic applications. Solar cells, where these materials are in greater demand, require a low cost production technology that will make the final product more affordable. Chemical bath deposition (CBD) a low cost growth technique capable of producing good quality thin film semiconductors over large area and at low temperature then becomes a suitable technology of choice. Heterogeneous reaction in a basic aqueous solution that is responsible for the II-VI compound film growth in CBD requires a metal complex. We have identified the stability constant (k) of the metal complex compatible with CBD growth mechanism to be about 106.9. This value is low enough to ensure that the substrate adsorbed complex relax for subsequent reaction with the chalcogen precursor to take place. It is also high enough to minimize the metal ion concentration in the bath participating in the precipitation of the bulk compounds. Homogeneous reaction that leads to precipitation in the reaction bath takes place because the solubility products of bulk II-VI compounds are very low. This reaction quickly depletes the bath of reactants, limit the film thickness, and degrade the film quality. While ZnS thin films are still hard to grow by CBD because of lack of suitable complexing agent, the homogeneous reaction still limits quality and thickness of both US and ZnS thin films. In this study, the zinc tetraammine complex ([Zn(NH3) 4]2+) with k = 108.9 has been forced to acquire its unsaturated form [Zn(NH3)3]2+ with a moderate k = 106.6 using hydrazine and nitrilotriacetate ion as complementary complexing agents and we have successfully grown ZnS thin films. We have also, minimized or eliminated the homogeneous reaction by using ammonium salt as a buffer and chemical bath with low reactant concentrations. These have allowed us to increase the saturation thickness of ZnS thin film by about 400% and raise that of US film

  7. Hydrogen production by hup(-) mutant and wild-type strains of Rhodobacter capsulatus from dark fermentation effluent of sugar beet thick juice in batch and continuous photobioreactors.

    PubMed

    Uyar, Basar; Gürgan, Muazzez; Özgür, Ebru; Gündüz, Ufuk; Yücel, Meral; Eroglu, Inci

    2015-10-01

    Photofermentative production of hydrogen is a promising and sustainable process; however, it should be coupled to dark fermentation to become cost effective. In order to integrate dark fermentation and photofermentation, the suitability of dark fermenter effluents for the photofermentative hydrogen production must be demonstrated. In this study, thermophilic dark fermenter effluent (DFE) of sugar beet thick juice was used as a substrate in photofermentation process to compare wild-type and uptake hydrogenase-deficient (hup (-)) mutant strains of Rhodobacter capsulatus by means of hydrogen production and biomass growth. The tests were conducted in small-scale (50 mL) batch and large-scale (4 L) continuous photobioreactors in indoor conditions under continuous illumination. In small scale batch conditions, maximum cell concentrations were 0.92 gdcw/L c and 1.50 gdcw/L c, hydrogen yields were 34 % and 31 %, hydrogen productivities were 0.49 mmol/(L c·h) and 0.26 mmol/(Lc·h), for hup (-) and wild-type cells, respectively. In large-scale continuous conditions, maximum cell concentrations were 1.44 gdcw/L c and 1.87 gdcw/L c, hydrogen yields were 48 and 46 %, and hydrogen productivities were 1.01 mmol/(L c·h) and 1.05 mmol/(L c·h), for hup (-) and wild-type cells, respectively. Our results showed that Rhodobacter capsulatus hup (-) cells reached to a lower maximum cell concentration but their hydrogen yield and productivity were in the same range or superior compared to the wild-type cells in both batch and continuous operating modes. The maximum biomass concentration, yield and productivity of hydrogen were higher in continuous mode compared to the batch mode with both bacterial strains. PMID:26164274

  8. Refinement of the high-resolution physical and genetic map of Rhodobacter capsulatus and genome surveys using blots of the cosmid encyclopedia.

    PubMed Central

    Fonstein, M; Koshy, E G; Nikolskaya, T; Mourachov, P; Haselkorn, R

    1995-01-01

    Cosmids from a library containing Rhodobacter capsulatus DNA fragments were previously ordered in two contigs: one corresponding to the chromosome and one to a 134 kb plasmid. This map contained 40 regions connected only by colony hybridization. To confirm the linkage and correct the map, the actual sizes of the overlaps were determined by blot-hybridization with Rhodobacter chromosomal DNA and by mapping of additional cosmids. Several revisions of the earlier map include single cosmid shifts and inversions. One additional gap in a cosmid contig was also found, raising the possibility that the chromosome is not a contiguous circle. About 2500 additional EcoRI,BamHI and HindIII restriction sites were added to the 560 EcoRV sites previously mapped onto the Rhodobacter chromosome, increasing the resolution of the physical map to the size of individual genes. Twenty-five new markers were located on the genetic map. The 48 markers now mapped represent nearly 300 genes and ORFs cloned from different species of Rhodobacter. The orientation of transcription of the four rrn operons was established using 16S rRNA- and 23S rRNA-specific probes and digestion with the rare-cutting enzyme, CeuI. Gel blots of 192 cosmids of the miniset of R.capsulatus digested with EcoRV were prepared. Such a hybridization template represents the whole genome cut into 560 DNA fragments varying in size from 0.4 to 25 kb. This template was used for high-resolution mapping of single genes, analysis of total genomic DNAs from related Rhodobacter strains and differentially expressed RNAs. Images PMID:7737133

  9. Overproduction of CcmG and CcmFHRc Fully Suppresses the c-Type Cytochrome Biogenesis Defect of Rhodobacter capsulatus CcmI-Null Mutants

    PubMed Central

    Sanders, Carsten; Deshmukh, Meenal; Astor, Doniel; Kranz, Robert G.; Daldal, Fevzi

    2005-01-01

    Gram-negative bacteria like Rhodobacter capsulatus use intertwined pathways to carry out the posttranslational maturation of c-type cytochromes (Cyts). This periplasmic process requires at least 10 essential components for apo-Cyt c chaperoning, thio-oxidoreduction, and the delivery of heme and its covalent ligation. One of these components, CcmI (also called CycH), is thought to act as an apo-Cyt c chaperone. In R. capsulatus, CcmI-null mutants are unable to produce c-type Cyts and thus sustain photosynthetic (Ps) growth. Previously, we have shown that overproduction of the putative heme ligation components CcmF and CcmHRc (also called Ccl1 and Ccl2) can partially bypass the function of CcmI on minimal, but not on enriched, media. Here, we demonstrate that either additional overproduction of CcmG (also called HelX) or hyperproduction of CcmF-CcmHRc is needed to completely overcome the role of CcmI during the biogenesis of c-type Cyts on both minimal and enriched media. These findings indicate that, in the absence of CcmI, interactions between the heme ligation and thioreduction pathways become restricted for sufficient Cyt c production. We therefore suggest that CcmI, along with its apo-Cyt chaperoning function, is also critical for the efficacy of holo-Cyt c formation, possibly via its close interactions with other components performing the final heme ligation steps during Cyt c biogenesis. PMID:15937187

  10. Bath Salts Abuse leading to New Onset Psychosis and Potential for Violence.

    PubMed

    John, Michelle E; Thomas-Rozea, Crystal; Hahn, David

    2014-06-20

    Background: Bath salts have recently emerged as a popular designer drug of abuse causing significant hazardous effects on mental health and physical health, resulting in public health legislation making its usage illegal in the United States.Objective: To educate mental health providers on the effects of the new designer drug bath salts, including its potential to cause psychosis and violence in patients.Method: This is a case report on a 40 year-old male with no past psychiatric history who presented with new onset psychosis and increased risk for violence after ingesting bath salts. In addition, a literature review was performed to summarize the documented effects of bath salts abuse and the current US public health legislation on bath salts.Results: The presented case illustrates a new onset, substance-induced psychotic disorder related to bath salts usage. The literature review explains the sympathomimetic reaction and the potential for psychotic symptoms.Discussion: To discuss the physical and psychological effects of bath salts, treatment options for bath salts abuse and US legislation by Ohio state law to current US federal law that bans production, sale, and possession of main substances found in bath salts.Conclusion: It is important for mental health providers to be aware of bath salts, understand the physical and psychiatric effects of bath salts and be familiar with current legislative policy banning its usage. Lastly, bath salts abuse should be in the differential diagnosis where psychosis is new onset or clinically incongruent with known primary presentation of a psychotic disorder. PMID:24951715

  11. A microscopic model for noise induced transport: Heat-bath nonlinearly driven by external white noise.

    PubMed

    Ghosh, Pradipta; Shit, Anindita; Chattopadhyay, Sudip; Chaudhuri, Jyotipratim Ray

    2011-03-01

    This work explores the observation that, even in the absence of a net externally applied bias, a symmetric homogeneous system coupled linearly to two heat baths is capable of producing unidirectional motion simply by nonlinearly driving one of the heat baths by an external Gaussian white noise. This is quite contrary to the traditional observation that, in order to obtain a net drift current, a state-dependent dissipation, which is a consequence of nonlinear system-bath coupling, is ubiquitous. PMID:21456831

  12. A microscopic model for noise induced transport: Heat-bath nonlinearly driven by external white noise

    SciTech Connect

    Ghosh, Pradipta; Shit, Anindita; Chattopadhyay, Sudip; Chaudhuri, Jyotipratim Ray

    2011-03-15

    This work explores the observation that, even in the absence of a net externally applied bias, a symmetric homogeneous system coupled linearly to two heat baths is capable of producing unidirectional motion simply by nonlinearly driving one of the heat baths by an external Gaussian white noise. This is quite contrary to the traditional observation that, in order to obtain a net drift current, a state-dependent dissipation, which is a consequence of nonlinear system-bath coupling, is ubiquitous.

  13. Electron transfer in a two-level system within a Cole-Davidson vitreous bath

    NASA Astrophysics Data System (ADS)

    Zarea, Mehdi; Ratner, Mark A.; Wasielewski, Michael R.

    2014-01-01

    We study electron transfer (ET) in a two level quantum system coupled to a glassy viscous bath. The bath is modeled by the Cole-Davidson (CD) spectral density. The ET in this model is compared to the ET in a normal Drude-Debye (DD) model. It is shown that at low temperatures and when the coupling to the bath is weak, the viscous bath preserves the quantum coherence for a longer time. However in the strong coupling regime, the tunneling rate is higher in the CD. In the classical high temperature limit the difference between the CD and DD models is negligible.

  14. Culture and long-term care: the bath as social service in Japan.

    PubMed

    Traphagan, John W

    2004-01-01

    A central feature of Japan's approach to community-based care of the elderly, including long-term home health care, is the emphasis on providing bath facilities. For mobile elders, senior centers typically provide a public bathing facility in which people can enjoy a relaxing soak along with friends who also visit the centers. In terms of in-home long-term care, visiting bath services are provided to assist family care providers with the difflcult task of bathing a frail or disabled elder--a task made more problematic as a result of the Japanese style of bathing. I argue that the bath, as social service, is a culturally shaped solution to a specific problem of elder care that arises in the Japanese context as a result of the importance of the bath in everyday life for Japanese. While the services may be considered specific to Japan, some aspects of bathing services, particularly the mobile bath service, may also have applicability in the United States. PMID:15792331

  15. Electron transfer in a two-level system within a Cole-Davidson vitreous bath

    SciTech Connect

    Zarea, Mehdi Ratner, Mark A.; Wasielewski, Michael R.

    2014-01-14

    We study electron transfer (ET) in a two level quantum system coupled to a glassy viscous bath. The bath is modeled by the Cole-Davidson (CD) spectral density. The ET in this model is compared to the ET in a normal Drude-Debye (DD) model. It is shown that at low temperatures and when the coupling to the bath is weak, the viscous bath preserves the quantum coherence for a longer time. However in the strong coupling regime, the tunneling rate is higher in the CD. In the classical high temperature limit the difference between the CD and DD models is negligible.

  16. Occurrence of thermotolerant Campylobacter spp. and adenoviruses in Finnish bathing waters and purified sewage effluents.

    PubMed

    Hokajärvi, Anna-Maria; Pitkänen, Tarja; Siljanen, Henri M P; Nakari, Ulla-Maija; Torvinen, Eila; Siitonen, Anja; Miettinen, Ilkka T

    2013-03-01

    A total of 50 Finnish bathing water samples and 34 sewage effluent samples originating from 17 locations were studied in the summers of 2006 and 2007. Campylobacter were present in 58% and adenoviruses in 12% of all bathing water samples; 53% of all sewage effluent samples were positive for Campylobacter spp. and 59% for adenoviruses. C. jejuni was the most common Campylobacter species found and human adenovirus serotype 41 was the most common identified adenovirus type. Bathing water temperature displayed a significant negative relationship with the occurrence of Campylobacter. One location had identical pulsed-field gel electrophoresis patterns of C. coli isolates in the bathing water and in sewage effluent, suggesting that sewage effluent was the source of C. coli at this bathing site. The counts of faecal indicator bacteria were not able to predict the presence of Campylobacter spp. or adenoviruses in the bathing waters. Thus the observed common presence of these pathogens in Finnish sewage effluents and bathing waters may represent a public health risk. The low water temperature in Finland may enhance the prevalence of Campylobacter in bathing waters. More attention needs to be paid to minimizing the concentrations of intestinal pathogens in bathing waters. PMID:23428555

  17. Quantum Otto engine using a single ion and a single thermal bath

    NASA Astrophysics Data System (ADS)

    Biswas, Asoka; Chand, Suman

    2016-05-01

    Quantum heat engines employ a quantum system as the working fluid, that gives rise to large work efficiency, beyond the limit for classical heat engines. Existing proposals for implementing quantum heat engines require that the system interacts with the hot bath and the cold bath (both modelled as a classical system) in an alternative fashion and therefore assumes ability to switch off the interaction with the bath during a certain stage of the heat-cycle. However, it is not possible to decouple a quantum system from its always-on interaction with the bath without use of complex pulse sequences. It is also hard to identify two different baths at two different temperatures in quantum domain, that sequentially interact with the system. Here, we show how to implement a quantum Otto engine without requiring to decouple the bath in a sequential manner. This is done by considering a single thermal bath, coupled to a single trapped ion. The electronic degree of freedom of the ion is chosen as a two-level working fluid while the vibrational degree of freedom plays the role of the cold bath. Measuring the electronic state mimics the release of heat into the cold bath. Thus, our model is fully quantum and exhibits very large work efficiency, asymptotically close to unity.

  18. Gauging a Quantum Heat Bath with Dissipative Landau-Zener Transitions

    SciTech Connect

    Wubs, Martijn; Kohler, Sigmund; Haenggi, Peter; Saito, Keiji; Kayanuma, Yosuke

    2006-11-17

    We calculate the exact Landau-Zener transition probabilities for a qubit with an arbitrary linear coupling to a bath at zero temperature. The final quantum state exhibits a peculiar entanglement between the qubit and the bath. In the special case of diagonal coupling, the bath does not influence the transition probability, whatever the speed of the Landau-Zener sweep. It is proposed to use Landau-Zener transitions to determine both the reorganization energy and the integrated spectral density of the bath. Possible applications include circuit QED and molecular nanomagnets.

  19. Nucleotide sequence and characterization of the Rhodobacter capsulatus hvrB gene: HvrB is an activator of S-adenosyl-L-homocysteine hydrolase expression and is a member of the LysR family.

    PubMed Central

    Buggy, J J; Sganga, M W; Bauer, C E

    1994-01-01

    Here we present the nucleotide sequence and characterization of two genes, hvrB and orf5, that are located in the regulatory gene cluster from Rhodobacter capsulatus. The hvrB gene, which encodes a protein with a predicted molecular mass of 32 kDa, is shown to be highly homologous to genes encoding members of the LysR family of bacterial transcriptional regulators. A chromosomal disruption of hvrB is shown to result in the failure to regulate expression from the nearby ahcY and orf5 genes in response to alterations in light intensity. We show by primer extension mapping that the 5' end of ahcY-specific mRNA defines a promoter region exhibiting sequence similarity to known R. capsulatus promoter elements. Our mutational analysis further demonstrates that hvrB autoregulates its own expression in vivo. Images PMID:8282711

  20. Heat-Bath Configuration Interaction: An Efficient Selected Configuration Interaction Algorithm Inspired by Heat-Bath Sampling.

    PubMed

    Holmes, Adam A; Tubman, Norm M; Umrigar, C J

    2016-08-01

    We introduce a new selected configuration interaction plus perturbation theory algorithm that is based on a deterministic analog of our recent efficient heat-bath sampling algorithm. This Heat-bath Configuration Interaction (HCI) algorithm makes use of two parameters that control the trade-off between speed and accuracy, one which controls the selection of determinants to add to a variational wave function and one which controls the selection of determinants used to compute the perturbative correction to the variational energy. We show that HCI provides an accurate treatment of both static and dynamic correlation by computing the potential energy curve of the multireference carbon dimer in the cc-pVDZ basis. We then demonstrate the speed and accuracy of HCI by recovering the full configuration interaction energy of both the carbon dimer in the cc-pVTZ basis and the strongly correlated chromium dimer in the Ahlrichs VDZ basis, correlating all electrons, to an accuracy of better than 1 mHa, in just a few minutes on a single core. These systems have full variational spaces of 3 × 10(14) and 2 × 10(22) determinants, respectively. PMID:27428771

  1. Promoters controlling expression of the alternative nitrogenase and the molybdenum uptake system in Rhodobacter capsulatus are activated by NtrC, independent of sigma54, and repressed by molybdenum.

    PubMed Central

    Kutsche, M; Leimkühler, S; Angermüller, S; Klipp, W

    1996-01-01

    The alternative nitrogenase of Rhodobacter capsulatus is expressed only under conditions of nitrogen and molybdenum depletion. The analysis of anfA-lacZ fusions demonstrated that this dual control occurred at the level of transcription of anfA, which encodes a transcriptional activator specific for the alternative nitrogenase. The anfA promoter was found to be activated under nitrogen-limiting conditions by NtrC in a sigma54-independent manner. In addition, anfA transcription was repressed by traces of molybdenum. This molybdenum-dependent repression of anfA was released in R. capsulatus mutants carrying either lesions in the high-affinity molybdenum uptake system (modABCD) or a double deletion of mopA and mopB, two genes encoding molybdenum-pterin-binding proteins. The expression of the molybdenum transport system itself was shown to be negatively regulated by molybdenum and, unexpectedly, to be also regulated by NtrC. This finding is in line with the presence of two tandemly arranged DNA motifs located in front of the R. capsulatus mopA-modABCD operon, which are homologous to R. capsulatus NtrC binding sites. Mapping of the transcriptional initiation sites of mopA and anfA revealed promoter sequences exhibiting significant homology to each other but no homology to known prokaryotic promoters. In addition, a conserved DNA sequence of dyad symmetry overlapping the transcriptional initiation sites of mopA and anfA was found. Deletions within this element resulted in molybdenum-independent expression of anfA, indicating that this DNA sequence may be the target of MopA/MopB-mediated repression. PMID:8606177

  2. Identification and sequence analysis of the hupR1 gene, which encodes a response regulator of the NtrC family required for hydrogenase expression in Rhodobacter capsulatus.

    PubMed Central

    Richaud, P; Colbeau, A; Toussaint, B; Vignais, P M

    1991-01-01

    The hupR1 gene from Rhodobacter capsulatus was cloned and sequenced. It can encode a protein of 53,843 Da which shares significant similarity with several transcriptional regulators and activates transcription of the structural hupSL genes of [NiFe]hydrogenase, as shown by the use of a translational fusion of lacZ with the hupSL promoter. A Hup- mutant having a point mutation in the hupR1 gene is described. PMID:1885559

  3. Open reading frame 5 (ORF5), encoding a ferredoxinlike protein, and nifQ are cotranscribed with nifE, nifN, nifX, and ORF4 in Rhodobacter capsulatus.

    PubMed

    Moreno-Vivian, C; Hennecke, S; Pühler, A; Klipp, W

    1989-05-01

    DNA sequence analysis of a 1,600-base-pair fragment located downstream of nifENX in nif region A of Rhodobacter capsulatus revealed two additional open reading frames (ORFs): ORF5, encoding a ferredoxinlike protein, and nifQ. The ferredoxinlike gene product contained two cysteine motifs, typical of ferredoxins coordinating two 4Fe-4S clusters, but the distance between these two motifs was unusual for low-molecular-weight ferredoxins. The R. capsulatus nifQ gene product shared a high degree of homology with Klebsiella pneumoniae and Azotobacter vinelandii NifQ, including a typical cysteine motif located in the C-terminal part. nifQ insertion mutants and also an ORF5-nifQ double deletion mutant showed normal diazotrophic growth only in the presence of high concentrations of molybdate. This demonstrated that the gene encoding the ferredoxinlike protein is not essential for nitrogen fixation. No NifA-activated consensus promoter could be found in the intergenic region between nifENX-ORF4 and ORF5-nifQ. Analyses of a nifQ-lacZYA fusion revealed that transcription of nifQ was initiated at a promoter in front of nifE. In contrast to other nitrogen-fixing organisms, R. capsulatus nifE, nifN, nifX, ORF4, ORF5, and nifQ were organized in one transcriptional unit. PMID:2708314

  4. Complementation of nitrogen-regulatory (ntr-like) mutations in Rhodobacter capsulatus by an Escherichia coli gene: cloning and sequencing of the gene and characterization of the gene product.

    PubMed Central

    Allibert, P; Willison, J C; Vignais, P M

    1987-01-01

    In vivo genetic engineering by R' plasmid formation was used to isolate an Escherichia coli gene that restored the Ntr+ phenotype to Ntr- mutants of the photosynthetic bacterium Rhodobacter capsulatus (formerly Rhodopseudomonas capsulata; J. F. Imhoff, H. G. Trüper, and N. Pfenning, Int. J. Syst. Bacteriol. 34:340-343, 1984). Nucleotide sequencing of the gene revealed no homology to the ntr genes of Klebsiella pneumoniae. Furthermore, hybridization experiments between the cloned gene and different F' plasmids indicated that the gene is located between 34 and 39 min on the E. coli genetic map and is therefore unlinked to the known ntr genes. The molecular weight of the gene product, deduced from the nucleotide sequence, was 30,563. After the gene was cloned in an expression vector, the gene product was purified. It was shown to have a pI of 5.8 and to behave as a dimer during gel filtration and on sucrose density gradients. Antibodies raised against the purified protein revealed the presence of this protein in R. capsulatus strains containing the E. coli gene, but not in other strains. Moreover, elimination of the plasmid carrying the E. coli gene from complemented strains resulted in the loss of the Ntr+ phenotype. Complementation of the R. capsulatus mutations by the E. coli gene therefore occurs in trans and results from the synthesis of a functional gene product. Images PMID:3025172

  5. Open reading frame 5 (ORF5), encoding a ferredoxinlike protein, and nifQ are cotranscribed with nifE, nifN, nifX, and ORF4 in Rhodobacter capsulatus.

    PubMed Central

    Moreno-Vivian, C; Hennecke, S; Pühler, A; Klipp, W

    1989-01-01

    DNA sequence analysis of a 1,600-base-pair fragment located downstream of nifENX in nif region A of Rhodobacter capsulatus revealed two additional open reading frames (ORFs): ORF5, encoding a ferredoxinlike protein, and nifQ. The ferredoxinlike gene product contained two cysteine motifs, typical of ferredoxins coordinating two 4Fe-4S clusters, but the distance between these two motifs was unusual for low-molecular-weight ferredoxins. The R. capsulatus nifQ gene product shared a high degree of homology with Klebsiella pneumoniae and Azotobacter vinelandii NifQ, including a typical cysteine motif located in the C-terminal part. nifQ insertion mutants and also an ORF5-nifQ double deletion mutant showed normal diazotrophic growth only in the presence of high concentrations of molybdate. This demonstrated that the gene encoding the ferredoxinlike protein is not essential for nitrogen fixation. No NifA-activated consensus promoter could be found in the intergenic region between nifENX-ORF4 and ORF5-nifQ. Analyses of a nifQ-lacZYA fusion revealed that transcription of nifQ was initiated at a promoter in front of nifE. In contrast to other nitrogen-fixing organisms, R. capsulatus nifE, nifN, nifX, ORF4, ORF5, and nifQ were organized in one transcriptional unit. PMID:2708314

  6. Bath salts and synthetic cathinones: an emerging designer drug phenomenon.

    PubMed

    German, Christopher L; Fleckenstein, Annette E; Hanson, Glen R

    2014-02-27

    Synthetic cathinones are an emerging class of designer drugs abused for psychostimulant and hallucinogenic effects similar to cocaine, methylenedioxymethamphetamine (MDMA), or other amphetamines. Abuse of synthetic cathinones, frequently included in products sold as 'bath salts', became prevalent in early 2009, leading to legislative classification throughout Europe in 2010 and schedule I classification within the United States in 2011. Recent pre-clinical and clinical studies indicate that dysregulation of central monoamine systems is a principal mechanism of synthetic cathinone action and presumably underlie the behavioral effects and abuse liability associated with these drugs. This review provides insight into the development of synthetic cathinones as substances of abuse, current patterns of their abuse, known mechanisms of their action and toxicology, and the benefits and drawbacks of their classification. PMID:23911668

  7. Skin hydration in nursing home residents using disposable bed baths.

    PubMed

    Gillis, Katrin; Tency, Inge; Roelant, Ella; Laureys, Sarina; Devriendt, Hendrik; Lips, Dirk

    2016-01-01

    The objective of this study was to evaluate a new way for applying bed baths and reducing the risk for dry skin by comparing the effect of two washing methods on skin hydration. A cluster randomized trial was conducted. Skin hydration was measured before and after implementation of disposable wash gloves, using a MoistureMeter SC at three skin sites. Total skin hydration did not differ between residents at the start of the study in both groups. After implementation, the post minus pre hydration scores were higher for the intervention group than the control group at all skin sites. However, the difference was only significant at cheek site. The use of disposable wash gloves does not increase the risk for dry skin in comparison with traditional washing methods. These results may encourage the introduction of disposable wash gloves as an innovation in daily skin care practice. PMID:26724816

  8. Bath salts and synthetic cathinones: An emerging designer drug phenomenon

    PubMed Central

    German, Christopher L.; Fleckenstein, Annette E.; Hanson, Glen R.

    2014-01-01

    The synthetic cathinones are an emerging class of designer drugs abused for psychostimulant and hallucinogenic effects similar to cocaine, methylenedioxymethamphetamine (MDMA), or other amphetamines. Abuse of synthetic cathinones, frequently included in products sold as ‘bath salts’, became prevalent in early 2009, leading to legislative classification throughout Europe in 2010 and schedule I classification within the United States in 2011. Recent pre-clinical and clinical studies indicate dysregulation of central monoamine systems are a principal mechanism of synthetic cathinone action and presumably underlie the behavioral effects and abuse liability associated with these drugs. This review provides insight into the development of synthetic cathinones as substances of abuse, current patterns of their abuse, known mechanisms of their action and toxicology, and the benefits and drawbacks of their classification. PMID:23911668

  9. Experimental bath engineering for quantitative studies of quantum control

    NASA Astrophysics Data System (ADS)

    Soare, A.; Ball, H.; Hayes, D.; Zhen, X.; Jarratt, M. C.; Sastrawan, J.; Uys, H.; Biercuk, M. J.

    2014-04-01

    We develop and demonstrate a technique to engineer universal unitary baths in quantum systems. Using the correspondence between unitary decoherence due to ambient environmental noise and errors in a control system for quantum bits, we show how a wide variety of relevant classical error models may be realized through in-phase or in-quadrature modulation on a vector signal generator producing a resonant carrier signal. We demonstrate our approach through high-bandwidth modulation of the 12.6-GHz carrier appropriate for trapped Yb171+ ions. Experiments demonstrate the reduction of coherent lifetime in the system in the presence of both engineered dephasing noise during free evolution and engineered amplitude noise during driven operations. In both cases, the observed reduction of coherent lifetimes matches well with quantitative models described herein. These techniques form the basis of a toolkit for quantitative tests of quantum control protocols, helping experimentalists characterize the performance of their quantum coherent systems.

  10. Asymptotic bound for heat-bath algorithmic cooling.

    PubMed

    Raeisi, Sadegh; Mosca, Michele

    2015-03-13

    The purity of quantum states is a key requirement for many quantum applications. Improving the purity is limited by fundamental laws of thermodynamics. Here, we are probing the fundamental limits for a natural approach to this problem, namely, heat-bath algorithmic cooling (HBAC). The existence of the cooling limit for HBAC techniques was proved by Schulman, Mor, and Weinstein. A bound for this value was found by Elias et al. and numerical testing supported the hypothesis that their bound may be the actual limit. A proof or disproof of whether their bound was the actual limit remained open for the past decade. Here, for the first time, we prove this limit. In the context of quantum thermodynamics, this corresponds to the maximum extractable work from the quantum system. We also establish, in the case of higher dimensional reset systems, how the performance of HBAC depends on the energy spectrum of the reset system. PMID:25815911

  11. Discovery, taxonomic distribution, and phenotypic characterization of a gene required for 3-methylhopanoid production

    PubMed Central

    Welander, Paula V.; Summons, Roger E.

    2012-01-01

    Hopanoids methylated at the C-3 position are a subset of bacterial triterpenoids that are readily preserved in modern and ancient sediments and in petroleum. The production of 3-methylhopanoids by extant aerobic methanotrophs and their common occurrence in modern and fossil methane seep communities, in conjunction with carbon isotope analysis, has led to their use as biomarker proxies for aerobic methanotrophy. In addition, these lipids are also produced by aerobic acetic acid bacteria and, lacking carbon isotope analysis, are more generally used as indicators for aerobiosis in ancient ecosystems. However, recent genetic studies have brought into question our current understanding of the taxonomic diversity of methylhopanoid-producing bacteria and have highlighted that a proper interpretation of methylhopanes in the rock record requires a deeper understanding of their cellular function. In this study, we identified and deleted a gene, hpnR, required for methylation of hopanoids at the C-3 position in the obligate methanotroph Methylococcus capsulatus strain Bath. Bioinformatics analysis revealed that the taxonomic distribution of HpnR extends beyond methanotrophic and acetic acid bacteria. Phenotypic analysis of the M. capsulatus hpnR deletion mutant demonstrated a potential physiological role for 3-methylhopanoids; they appear to be required for the maintenance of intracytoplasmic membranes and cell survival in late stationary phase. Therefore, 3-methylhopanoids may prove more useful as proxies for specific environmental conditions encountered during stationary phase rather than a particular bacterial group. PMID:22826256

  12. Discovery, taxonomic distribution, and phenotypic characterization of a gene required for 3-methylhopanoid production.

    PubMed

    Welander, Paula V; Summons, Roger E

    2012-08-01

    Hopanoids methylated at the C-3 position are a subset of bacterial triterpenoids that are readily preserved in modern and ancient sediments and in petroleum. The production of 3-methylhopanoids by extant aerobic methanotrophs and their common occurrence in modern and fossil methane seep communities, in conjunction with carbon isotope analysis, has led to their use as biomarker proxies for aerobic methanotrophy. In addition, these lipids are also produced by aerobic acetic acid bacteria and, lacking carbon isotope analysis, are more generally used as indicators for aerobiosis in ancient ecosystems. However, recent genetic studies have brought into question our current understanding of the taxonomic diversity of methylhopanoid-producing bacteria and have highlighted that a proper interpretation of methylhopanes in the rock record requires a deeper understanding of their cellular function. In this study, we identified and deleted a gene, hpnR, required for methylation of hopanoids at the C-3 position in the obligate methanotroph Methylococcus capsulatus strain Bath. Bioinformatics analysis revealed that the taxonomic distribution of HpnR extends beyond methanotrophic and acetic acid bacteria. Phenotypic analysis of the M. capsulatus hpnR deletion mutant demonstrated a potential physiological role for 3-methylhopanoids; they appear to be required for the maintenance of intracytoplasmic membranes and cell survival in late stationary phase. Therefore, 3-methylhopanoids may prove more useful as proxies for specific environmental conditions encountered during stationary phase rather than a particular bacterial group. PMID:22826256

  13. Identification of a bacterial di-haem cytochrome c peroxidase from Methylomicrobium album BG8.

    PubMed

    Karlsen, O A; Larsen, Ø; Jensen, H B

    2010-09-01

    The nucleotide sequence of an open reading frame (corB) downstream of the copper-repressible CorA-encoding gene of the methanotrophic bacterium Methylomicrobium album BG8 was obtained by restriction enzyme digestion and inverse PCR. The amino acid sequence deduced from this gene showed significant sequence similarity to the surface-associated di-haem cytochrome c peroxidase (SACCP) previously isolated from Methylococcus capsulatus (Bath), including both c-type haem-binding motifs. Homology analysis placed this protein, phylogenetically, within the subfamily containing the M. capsulatus SACCP of the bacterial di-haem cytochrome c peroxidase (BCCP) family of proteins. Immunospecific recognition confirmed synthesis of the M. album CorB as a protein non-covalently associated with the outer membrane and exposed to the periplasm. corB expression is regulated by the availability of copper ions during growth and the protein is most abundant in M. album when grown at a low copper-to-biomass ratio, indicating an important physiological role of CorB under these growth conditions. corB was co-transcribed with the gene encoding CorA, constituting a copper-responding operon, which appears to be under the control of a sigma(54)-dependent promoter. M. album CorB is the second isolated member of the recently described subfamily of the BCCP family of proteins. So far, these proteins have only been described in methanotrophic bacteria. PMID:20576687

  14. Analyses of Methanobactin, a Novel Copper-Binding Compound, or Chalkophore, from Methanotrophs

    NASA Astrophysics Data System (ADS)

    Semrau, J. D.; Yoon, S.; Dispirito, A.; Kraemer, S.

    2009-12-01

    Methanotrophs, cells that utilize methane as their sole carbon and energy source are known to have high requirements for copper. Many of these cells are known to synthesize a copper-chelating agent, or chalkophore, termed methanobactin that appears integral to copper uptake. Structural analyses indicate that methanobactin is a small chromopeptide that binds copper utilizing two alkylidene oxazolone rings. Using a suite of spectrophotemetric and calorimetry analyses, it was discovered that methanobactin preferentially binds copper, but also binds other metals, including gold, forming gold nanoparticles. To screen methanobactin production by methanotrophs, as well as to determine if other cells make chalkophores, a plate assay developed from the Chromo Azurol S (CAS) assay for siderophore production, was modified. In the standard CAS assay, a color change in CAS plate is observed as iron (III) ion weakly bound to CAS is taken up by siderophores. In our modified assay, iron (III) chloride of the initial CAS solution was substituted with copper(II) chloride. Assay results indicated that of the four tested methanotrophs (Methylomicrobium album BG8, Methylosinus trichosporium OB3b, Methylococcus capsulatus Bath, and Methylocystis parvus OBBP), only M. album BG8, M. trichosporium OB3b, and M. capsulatus produced methanobactin, while M. parvus OBBP did not. The assay can be easily adopted for detection of chalkophores in other microorganisms and detection as well as for screening of putative mutants of chalkophore synthesis.

  15. Soluble methane monooxygenase component B gene probe for identification of methanotrophs that rapidly degrade trichloroethylene

    SciTech Connect

    Hsienchyang Tsien; Hanson, R.S. )

    1992-03-01

    Restriction fragment length polymorphisms, Western blot (immunoblot) analysis, and fluorescence-labelled signature probes were used for the characterization of methanotrophic bacteria as well as for the identification of methanotrophs which contained the soluble methane monooxygenase (MMO) gene and were able to degrade trichloroethylene (TCE). The gene encoding a soluble MMO component B protein from Methylosinus trichosporium OB3b was cloned. It contained a 2.2-kb EcoRI fragment. With this cloned component B gene as probe, methanotroph types I, II, and X and environmental and bioreactor samples were screened for the presence of the gene encoding soluble MMO. Among twelve pure or mixed cultures, DNA fragments of seven methanotrophs hybridized with the soluble MMO B gene probe. When grown in media with limited copper, all of these bacteria degraded TCE. All of them are type II methanotrophs. The soluble MMO component B gene of the type X methanotroph, Methylococcus capsulatus Bath, did not hybridize to the M. trichosporium OB3b soluble MMO component B gene probe, although M. capsulatus Baath also produces a soluble MMO.

  16. Discovery, taxonomic distribution, and phenotypic characterization of a gene required for 3-methylhopanoid production

    NASA Astrophysics Data System (ADS)

    Welander, Paula V.; Summons, Roger E.

    2012-08-01

    Hopanoids methylated at the C-3 position are a subset of bacterial triterpenoids that are readily preserved in modern and ancient sediments and in petroleum. The production of 3-methylhopanoids by extant aerobic methanotrophs and their common occurrence in modern and fossil methane seep communities, in conjunction with carbon isotope analysis, has led to their use as biomarker proxies for aerobic methanotrophy. In addition, these lipids are also produced by aerobic acetic acid bacteria and, lacking carbon isotope analysis, are more generally used as indicators for aerobiosis in ancient ecosystems. However, recent genetic studies have brought into question our current understanding of the taxonomic diversity of methylhopanoid-producing bacteria and have highlighted that a proper interpretation of methylhopanes in the rock record requires a deeper understanding of their cellular function. In this study, we identified and deleted a gene, hpnR, required for methylation of hopanoids at the C-3 position in the obligate methanotroph Methylococcus capsulatus strain Bath. Bioinformatics analysis revealed that the taxonomic distribution of HpnR extends beyond methanotrophic and acetic acid bacteria. Phenotypic analysis of the M. capsulatus hpnR deletion mutant demonstrated a potential physiological role for 3-methylhopanoids; they appear to be required for the maintenance of intracytoplasmic membranes and cell survival in late stationary phase. Therefore, 3-methylhopanoids may prove more useful as proxies for specific environmental conditions encountered during stationary phase rather than a particular bacterial group.

  17. [Interference therapy and radon baths in the combined treatment of patients with reflex cervicobrachial syndromes].

    PubMed

    Gorbunov, F E; Semenistaia, S V

    1998-01-01

    Patients with cervicobrachialgic syndromes on interference therapy, exercise treatment, massage of the cervical collar region received balneotherapy. 42 of them took water baths, 39 took dry air radon baths. These complexes proved effective in cervicobrachialgic syndromes, the effect being slightly dependent on the clinical symptoms of the disease. PMID:9987978

  18. Transport coefficients of a quantum system interacting with a squeezed heat bath

    SciTech Connect

    Kalandarov, Sh. A.; Adamian, G. G.; Kanokov, Z.; Antonenko, N. V.

    2006-07-15

    The analytical expressions for the time-dependent friction and diffusion coefficients are presented for the case of coupling in coordinates between the collective subsystem and a squeezed heat bath. The effects of initial phase-sensitive and -insensitive correlations of the heat bath on the diffusion coefficients, fluctuations, and decoherence are studied. The interplay between friction and decoherence is discussed.

  19. Copper Sulfate Foot Baths on Dairies and Crop Toxicities – What are the Risks?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A rising concern with the application of dairy wastes to agricultural fields is the accumulation of copper (Cu) in the soil. Copper sulfate (CuSO4) from cattle foot baths are washed out of dairy barns and into wastewater lagoons. The addition of CuSO4 baths has been reported to increase Cu concent...

  20. Quantum thermodynamic functions for an oscillator coupled to a heat bath

    NASA Astrophysics Data System (ADS)

    Ford, G. W.; O'Connell, R. F.

    2007-04-01

    Small systems (of interest in the areas of nanophysics, quantum information, etc.) are particularly vulnerable to environmental effects. Thus, we determine various thermodynamic functions for an oscillator in an arbitrary heat bath at arbitrary temperatures. Explicit results are presented for the most commonly discussed heat bath models: Ohmic, single relaxation time, and blackbody radiation.

  1. 78 FR 73506 - Submission for OMB Review; Comment Request: Infant Bath Seats

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-06

    ... modifications to reduce further the risk of injury associated with infant bath seats. 75 FR 31691. On July 31, 2012, the Commission adopted the revised ASTM standard for infant bath seats, ASTM F1967-11a. 77 FR....gov . SUPPLEMENTARY INFORMATION: In the Federal Register of August 30, 2013 (78 FR 53734),...

  2. CURRENT AND EMERGING TECHNOLOGIES FOR EXTENDING THE LIFETIME OF ELECTROLESS NICKEL PLATING BATHS

    EPA Science Inventory

    The waste treatment and rejuvenation of spent electroless nickel baths has attracted a considerable amount of interest from electroplating shops, electroless nickel suppliers, universities and regulatory agencies due to the finite life of the baths and the associated waste that t...

  3. Copper sulfate foot baths on dairies and crop toxicities – What are the risks?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A rising concern with the application of dairy wastes to agricultural fields is the accumulation of copper (Cu) in the soil. Copper sulfate (CuSO4) from cattle foot baths are washed out of dairy barns and into wastewater lagoons. The addition of CuSO4 baths has been reported to increase Cu concent...

  4. [An outbreak of legionellosis in a new facility of hot spring bath in Hiuga City].

    PubMed

    Yabuuchi, Eiko; Agata, Kunio

    2004-02-01

    Following cerebrating ceremony in 20 June 2002, for the completion of Hiuga Sun-Park Hot Spring Bath "Ofunade-no-Yu" facilities, Miyazaki Prefecture, Kyushu Island, 200 neighbors were invited each day to experience bathing on 20 and 21 June. The Bath "Ofunade-no-Yu" officially opened on 1 July 2002. On 18 July, Hiuga Health Center was informed that 3 suspected Legionella pneumonia patients in a hospital and all of them have bathing history of "Ofunade-no-Yu". Health Center officers notified Hiuga City, the main proprietor of the Bath business, that on-site inspection on sanitary managements will be done next day and requested the City to keep the bath facilities as they are. On 19 July, Health Center officers collected bath water from seven places and recommended voluntary-closing of "Ofunade-no-Yu" business. Because of various reasons, Hiuga City did not accept the recommendation and continued business up to 23 July. Because Legionella pneumophila serogroup 1 strains from 4 patients' sputa and several bath water specimens were determined genetically similar by Pulsed Field Gel Electrophoresis of Sfi I-cut DNA. "Ofunede-no-Yu" was regarded as the source of infection of this outbreak. On 24 July, "Ofunade-no-Yu" accepted the Command to prohibit the business. Among 19,773 persons who took the bath during the period from 20 June to 23 July, 295 became ill, and 7 died. Among them, 34 were definitely diagnosed as Legionella pneumonia due to L. pneumophila SG 1, by either one or two tests of positive sputum culture, Legionella-specific urinary antigen, and significant rise of serum antibody titer against L. pneumophila SG 1. In addition to the 8 items shown by Miyazaki-Prefecture Investigation Committee as the cause of infection. Hiuga City Investigation Committee pointed out following 3 items: 1) Insufficient knowledge and understanding of stuffs on Legionella and legionellosis; 2) Residual water in tubing system after trial runs might lead multiplication of legionellae

  5. Effect of chlorhexidine bathing in preventing infections and reducing skin burden and environmental contamination: A review of the literature.

    PubMed

    Donskey, Curtis J; Deshpande, Abhishek

    2016-05-01

    Chlorhexidine bathing is effective in reducing levels of pathogens on skin. In this review, we examine the evidence that chlorhexidine bathing can prevent colonization and infection with health care-associated pathogens and reduce dissemination to the environment and the hands of personnel. The importance of education and monitoring of compliance with bathing procedures is emphasized in order to optimize chlorhexidine bathing in clinical practice. PMID:27131130

  6. Homologues of Genetic Transformation DNA Import Genes Are Required for Rhodobacter capsulatus Gene Transfer Agent Recipient Capability Regulated by the Response Regulator CtrA

    PubMed Central

    Brimacombe, Cedric A.; Ding, Hao; Johnson, Jeanette A.

    2015-01-01

    ABSTRACT Gene transfer agents (GTAs) morphologically resemble small, double-stranded DNA (dsDNA) bacteriophages; however, their only known role is to package and transfer random pieces of the producing cell genome to recipient cells. The best understood GTA is that of Rhodobacter capsulatus, termed RcGTA. We discovered that homologues of three genes involved in natural transformation in other bacteria, comEC, comF, and comM, are essential for RcGTA-mediated gene acquisition. This paper gives genetic and biochemical evidence that RcGTA-borne DNA entry into cells requires the ComEC and ComF putative DNA transport proteins and genetic evidence that putative cytoplasmic ComM protein of unknown function is required for recipient capability. Furthermore, the master regulator of RcGTA production in <1% of a cell population, CtrA, which is also required for gene acquisition in recipient cells, is expressed in the vast majority of the population. Our results indicate that RcGTA-mediated gene transfer combines key aspects of two bacterial horizontal gene transfer mechanisms, where donor DNA is packaged in transducing phage-like particles and recipient cells take up DNA using natural transformation-related machinery. Both of these differentiated subsets of a culture population, donors and recipients, are dependent on the same response regulator, CtrA. IMPORTANCE Horizontal gene transfer (HGT) is a major driver of bacterial evolution and adaptation to environmental stresses. Traits such as antibiotic resistance or metabolic properties can be transferred between bacteria via HGT; thus, HGT can have a tremendous effect on the fitness of a bacterial population. The three classically described HGT mechanisms are conjugation, transformation, and phage-mediated transduction. More recently, the HGT factor GTA was described, where random pieces of producing cell genome are packaged into phage-like particles that deliver DNA to recipient cells. In this report, we show that transport of

  7. Cardiac bidomain bath-loading effects during arrhythmias: interaction with anatomical heterogeneity.

    PubMed

    Bishop, Martin J; Vigmond, Edward; Plank, Gernot

    2011-12-21

    Cardiac tissue is always surrounded by conducting fluid, both in vivo (blood) and in experimental preparations (Tyrode's solution), which acts to increase conduction velocity (CV) close to the tissue-fluid interface, inducing transmural wavefront curvature. Despite its potential importance, computer modeling studies focused on arrhythmia mechanisms have previously not accounted for these bath-loading effects. Here, we investigate the increase in CV and concomitant change in transmural wavefront profiles upon both propagation and arrhythmia dynamics within models of differing anatomical complexity. In simplified slab models, in absence of transmural fiber rotation, bath-loading induced transmural wavefront curvature dominates, significantly increasing arrhythmia complexity compared to no bath. In the presence of fiber rotation, bath-loading effects are less striking and depend upon propagation direction: the bath accentuates natural concave curvature caused by transmurally rotating fibers, but attenuates convex curvature, which negates overall impact upon arrhythmia complexity. Finally, we demonstrate that the high degree of anatomical complexity within whole ventricular models modulates bath-loading induced transmural wavefront curvature. However, key is the increased surface CV that dramatically reduces both arrhythmia inducibility and resulting complexity by increasing wavelength and reducing the available excitable gap. Our findings highlight the importance of including bath-loading effects during arrhythmia mechanism investigations, which could have implications for interpreting and comparing simulation results with experimental data where such effects are inherently present. PMID:22208185

  8. Locomotor stimulant and discriminative stimulus effects of 'bath salt' cathinones.

    PubMed

    Gatch, Michael B; Taylor, Cynthia M; Forster, Michael J

    2013-09-01

    A number of psychostimulant-like cathinone compounds are being sold as 'legal' alternatives to methamphetamine or cocaine. The purpose of these experiments was to determine whether cathinone compounds stimulate motor activity and have discriminative stimulus effects similar to those of cocaine and/or methamphetamine. 3,4-Methylenedioxypyrovalerone (MDPV), methylone, mephedrone, naphyrone, flephedrone, and butylone were tested for locomotor stimulant effects in mice and subsequently for substitution in rats trained to discriminate cocaine (10 mg/kg, intraperitoneally) or methamphetamine (1 mg/kg, intraperitoneally) from saline. All compounds fully substituted for the discriminative stimulus effects of cocaine and methamphetamine. Several commonly marketed cathinones produce discriminative stimulus effects comparable with those of cocaine and methamphetamine, which suggests that these compounds are likely to have similar abuse liabilities. MDPV and naphyrone produced locomotor stimulant effects that lasted much longer than those of cocaine or methamphetamine and therefore may be of particular concern, particularly because MDPV is one of the most commonly found substances associated with emergency room visits because of adverse effects of taking 'bath salts'. PMID:23839026

  9. Quantum Thermal Bath for Path Integral Molecular Dynamics Simulation.

    PubMed

    Brieuc, Fabien; Dammak, Hichem; Hayoun, Marc

    2016-03-01

    The quantum thermal bath (QTB) method has been recently developed to account for the quantum nature of the nuclei by using standard molecular dynamics (MD) simulation. QTB-MD is an efficient but approximate method when dealing with strongly anharmonic systems, while path integral molecular dynamics (PIMD) gives exact results but in a huge amount of computation time. The QTB and PIMD methods have been combined in order to improve the PIMD convergence or correct the failures of the QTB-MD technique. Therefore, a new power spectral density of the random force within the QTB has been developed. A modified centroid-virial estimator of the kinetic energy, especially adapted to QTB-PIMD, has also been proposed. The method is applied to selected systems: a one-dimensional double-well system, a ferroelectric phase transition, and the position distribution of an hydrogen atom in a fuel cell material. The advantage of the QTB-PIMD method is its ability to give exact results with a more reasonable computation time for strongly anharmonic systems. PMID:26799437

  10. Mechanisms of Carrier Transport Induced by a Microswimmer Bath

    SciTech Connect

    Kaiser, Andreas; Sokolov, Andrey; Aranson, Igor S.; Lowen, Hartmut

    2015-04-01

    Recently, it was found that a wedgelike microparticle (referred to as ”carrier”) which is only allowed to translate but not to rotate exhibits a directed translational motion along the wedge cusp if it is exposed to a bath of microswimmers. Here we model this effect in detail by resolving the microswimmers explicitly using interaction models with different degrees of mutual alignment. Using computer simulations we study the impact of these interactions on the transport efficiency of V-shaped carrier. We show that the transport mechanisms itself strongly depends on the degree of alignment embodied in the modelling of the individual swimmer dynamics. For weak alignment, optimal carrier transport occurs in the turbulent microswimmer state and is induced by swirl depletion inside the carrier. For strong aligning interactions, optimal transport occurs already in the dilute regime and is mediated by a polar cloud of swimmers in the carrier wake pushing the wedge-particle forward. We also demonstrate that the optimal shape of the carrier leading to maximal transport speed depends on the kind of interaction model used.

  11. Laser cooling of a harmonic oscillator's bath with optomechanics

    NASA Astrophysics Data System (ADS)

    Xu, Xunnong; Taylor, Jacob

    Thermal noise reduction in mechanical systems is a topic both of fundamental interest for studying quantum physics at the macroscopic level and for application of interest, such as building high sensitivity mechanics based sensors. Similar to laser cooling of neutral atoms and trapped ions, the cooling of mechanical motion by radiation pressure can take single mechanical modes to their ground state. Conventional optomechanical cooling is able to introduce additional damping channel to mechanical motion, while keeping its thermal noise at the same level, and as a consequence, the effective temperature of the mechanical mode is lowered. However, the ratio of temperature to quality factor remains roughly constant, preventing dramatic advances in quantum sensing using this approach. Here we propose an efficient scheme for reducing the thermal load on a mechanical resonator while improving its quality factor. The mechanical mode of interest is assumed to be weakly coupled to its heat bath but strongly coupled to a second mechanical mode, which is cooled by radiation pressure coupling to a red detuned cavity field. We also identify a realistic optomechanical design that has the potential to realize this novel cooling scheme. Joint Center for Quantum Information and Computer Science, University of Maryland, College Park, MD 20742, USA.

  12. Mephedrone (‘bath salt’) pharmacology: insights from invertebrates

    PubMed Central

    Ramoz, Leda; Lodi, Sabah; Bhatt, Parth; Reitz, Allen B.; Tallarida, Chris; Tallarida, Ronald J.; Raffa, Robert B.; Rawls, Scott M.

    2014-01-01

    Psychoactive bath salts (also called meph, drone, meow meow, m-CAT, bounce, bubbles, mad cow, etc.) contain a substance called mephedrone (4-methylcathinone) that may share psychostimulant properties with amphetamine and cocaine. However, there are only limited studies of the neuropharmacological profile of mephedrone. The present study used an established invertebrate (planarian) assay to test the hypothesis that acute and repeated mephedrone exposure produces psychostimulant-like behavioral effects. Acute mephedrone administration (50 – 1000 µM) produced stereotyped movements that were attenuated by a dopamine receptor antagonist (SCH 23390) (0.3 µM). Spontaneous discontinuation of mephedrone exposure (1, 10 µM) (60 min) resulted in an abstinence-induced withdrawal response (i.e., reduced motility). In place conditioning experiments, planarians in which mephedrone (100, 500 µM) was paired with the non-preferred environment during conditioning displayed a shift in preference upon subsequent testing. These results suggest that mephedrone produces three behavioral effects associated with psychostimulant drugs, namely dopamine-sensitive stereotyped movements, abstinence-induced withdrawal, and environmental place conditioning. PMID:22300981

  13. Nonequilibrium Response of Nanosystems Coupled to Driven Quantum Baths.

    PubMed

    Grabert, Hermann; Nalbach, Peter; Reichert, Joscha; Thorwart, Michael

    2016-06-01

    Commonly, nanosystems are characterized by their response to time-dependent external fields in the presence of inevitable environmental fluctuations. The direct impact of the external driving on the environment is generally neglected. While this approach is satisfactory for macroscopic systems, on the nanoscale, an interaction of external fields with the environment is often unavoidable on principle. We extend the standard linear response theory of quantum dissipative systems to strongly driven baths. Significant modifications are found for two paradigm examples. First, we evaluate the polarizability of a molecule immersed in a strongly polarizable medium that responds to terahertz radiation. We find an increase of the molecular polarizability by about 30%. Second, we determine the response of a semiconductor quantum dot in close proximity to a metallic nanoparticle. Both are placed in a polarizable medium and exposed to electromagnetic irradiation. We show that the response of the quantum dot is qualitatively modified by the driven nanoparticle, including the generation of an additional channel of stimulated emission. PMID:27176818

  14. Risk factors of sudden death in the Japanese hot bath in the senior population.

    PubMed

    Chiba, Takashi; Yamauchi, Misa; Nishida, Naoki; Kaneko, Taeko; Yoshizaki, Katsuaki; Yoshioka, Naofumi

    2005-05-10

    A series of experiments were carried out to clarify the cause of death and the risk factors related to sudden death in the Japanese senior population while bathing in a Japanese style "hot bath." The biodynamic changes while bathing were carefully monitored under actual bathing situations occurring in both the winter and summer seasons. We observed double product (DP), total peripheral blood vessel resistance (TPR), cardiac output (CO), and blood vessel compliance (COMP) by measuring blood pressure, heart rate, pulse wave, and electrocardiogram (ECG). The finding of a high level of DP in the elderly suggests that more myocardial oxygen consumption is needed than for young adults, particularly in subjects with arrhythmia. Although the values for TPR and CO changed somewhat during bathing, the changes were considered normal and to be expected. However, more significant and substantial changes were observed during the winter experiment than during the summer experiment, no doubt owing to lower temperature of the bathing room. The value of COMP did not vary significantly between winter and summer subjects. Twelve subjects in the elderly developed ECG changes while bathing such as supraventricular extrasystole or ventricular tachycardia. No clinical significance was found in the biochemical analyses of the blood obtained before and after bathing. In conclusion, some subjects in the elderly showed risky changes in the above parameters and ECG, factors which may partially explain some of the causes of the many reported cases of lapse of consciousness and unexpected sudden death in the elderly while bathing especially in the winter season. Cold climate, hot water immersion, and hydrostatic pressure may affect their physiological compensation along with existing of coronary stenosis or weakness of respiratory function as a normal consequence of advanced age. PMID:15749356

  15. The effects of daily bathing on symptoms of patients with bronchial asthma

    PubMed Central

    Arimoto, Yoshihito; Homma, Chie; Takeoka, Shinjiro; Fukusumi, Munehisa; Mouri, Atsuto; Hamamoto, Yoichiro

    2016-01-01

    Background The influence of bathing in asthma patients is not yet fully known. Objective We conducted an observational study to investigate changes in symptoms and their degree by bathing in asthmatic patients. Methods A questionnaire focusing on ever experienced bathing-induced symptom changes and their degree, as well as contributing factors, was designed and administered to asthmatic patients in the outpatient department of our institute between January 2012 and November 2013. Results Two hundred fifteen cases were recruited. In 60 cases (27.9%), asthmatic symptoms appeared, including 20 cases of chest discomfort (33.3%), 19 cases of cough (31.7%), and 21 cases of wheezing (35.0%). The triggering factors included vapor inhalation (32 cases, 53.3%), hydrostatic pressure on the thorax due to body immersion in the bathtub (26 cases, 43.3%), and sudden change of air temperature (16 cases, 26.7%). Thirty-eight cases (17.7%) experienced improvement in active asthmatic symptoms by bathing. Vapor inhalation was the most common contributing factor (34 cases, 89.5%), followed by warming of the whole body (13 cases, 34.2%). There was no relationship between asthma severity and the appearance of bathing-induced symptoms or improvement of active asthmatic symptoms by bathing. Conclusion The effects of bathing in asthmatic patients widely differed from patient to patient and their etiology includes several factors. For those who suffer from bathing-induced asthma symptoms, preventive methods, such as premedication with bronchodilators before bathing, should be established. This study is registered in the University Hospital Medical Information Network (UMIN) clinical trials registry in Japan with the registration number UMIN000015641. PMID:27141485

  16. On the operation of machines powered by quantum non-thermal baths

    NASA Astrophysics Data System (ADS)

    Niedenzu, Wolfgang; Gelbwaser-Klimovsky, David; Kofman, Abraham G.; Kurizki, Gershon

    2016-08-01

    Diverse models of engines energised by quantum-coherent, hence non-thermal, baths allow the engine efficiency to transgress the standard thermodynamic Carnot bound. These transgressions call for an elucidation of the underlying mechanisms. Here we show that non-thermal baths may impart not only heat, but also mechanical work to a machine. The Carnot bound is inapplicable to such a hybrid machine. Intriguingly, it may exhibit dual action, concurrently as engine and refrigerator, with up to 100% efficiency. We conclude that even though a machine powered by a quantum bath may exhibit an unconventional performance, it still abides by the traditional principles of thermodynamics.

  17. Approach to equilibrium in models of a system in contact with a heat bath

    SciTech Connect

    Goldstein, S.; Lebowitz, J.L.; Ravishankar, K.

    1986-04-01

    We investigate simple model systems in contact with an infinite heat bath. The former consists of a finite number of particles in a bounded region ..lambda.. of R/sup d/, d = 1, 2. The heat baths are infinite particle systems which can penetrate ..lambda.. and interact with the system via elastic collisions. Outside ..lambda.. the particles move freely and have a Gibbs probability measure prior to entering ..lambda... We show that starting from almost any initial configuration, the system approaches, as t ..-->.. infinity, the appropriate Gibbs distribution. The combined system plus bath is Bernoulli.

  18. Portable oil bath for high-accuracy resistance transfer and maintenance

    NASA Astrophysics Data System (ADS)

    Shiota, Fuyuhiko

    1999-10-01

    A portable oil bath containing one standard resistor for high-accuracy resistance transfer and maintenance was developed and operated for seven years in the National Research Laboratory of Metrology. The aim of the bath is to save labor and apparatus for high-accuracy resistance transfer and maintenance by consistently keeping the standard resistor in an optimum environmental condition. The details of the prototype system, including its performance, are described together with some suggestions for a more practical bath design, which adopts the same concept.

  19. Carotenoid biosynthesis in bacteria: In vitro studies of a crt/bch transcription factor from Rhodobacter capsulatus and carotenoid enzymes from Erwinia herbicola

    SciTech Connect

    O`Brien, D.A.

    1992-11-01

    A putative transcription factor in Rhodobactor capsulatus which binds upstream of the crt and bch pigment biosynthesis operons and appears to play a role in the adaptation of the organism from the aerobic to the anaerobic-photosynthetic growth mode was characterized. Chapter 2 describes the identification of this factor through an in vitro mobility shift assay, as well as the determination of its binding properties and sequence specificity. Chapter 3 focuses on the isolation of this factor. Biochemistry of later carotenoid biosynthesis enzymes derived from the non-photosynthetic bacterium, Erwinia herbicola. Chapter 4 describes the separate overexpression and in vitro analysis of two enzymes involved in the main sequence of the carotenoid biosynthesis pathway, lycopene cyclase and 5-carotene hydroxylase. Chapter 5 examines the overexpression and enzymology of functionally active zeaxanthin glucosyltransferase, an enzyme which carries out a more unusual transformation, converting a carotenoid into its more hydrophilic mono- and diglucoside derivatives. In addition, amino acid homology with other glucosyltransferases suggests a putative binding site for the UDP-activated glucose substrate.

  20. NifA- and CooA-Coordinated cowN Expression Sustains Nitrogen Fixation by Rhodobacter capsulatus in the Presence of Carbon Monoxide

    PubMed Central

    Hoffmann, Marie-Christine; Pfänder, Yvonne; Fehringer, Maria; Narberhaus, Franz

    2014-01-01

    Rhodobacter capsulatus fixes atmospheric dinitrogen via two nitrogenases, Mo- and Fe-nitrogenase, which operate under different conditions. Here, we describe the functions in nitrogen fixation and regulation of the rcc00574 (cooA) and rcc00575 (cowN) genes, which are located upstream of the structural genes of Mo-nitrogenase, nifHDK. Disruption of cooA or cowN specifically impaired Mo-nitrogenase-dependent growth at carbon monoxide (CO) concentrations still tolerated by the wild type. The cooA gene was shown to belong to the Mo-nitrogenase regulon, which is exclusively expressed when ammonium is limiting. Its expression was activated by NifA1 and NifA2, the transcriptional activators of nifHDK. AnfA, the transcriptional activator of Fe-nitrogenase genes, repressed cooA, thereby counteracting NifA activation. CooA activated cowN expression in response to increasing CO concentrations. Base substitutions in the presumed CooA binding site located upstream of the cowN transcription start site abolished cowN expression, indicating that cowN regulation by CooA is direct. In conclusion, a transcription factor-based network controls cowN expression to protect Mo-nitrogenase (but not Fe-nitrogenase) under appropriate conditions. PMID:25070737