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Sample records for pyrococcus furiosus exhibits

  1. Operon prediction in Pyrococcus furiosus

    PubMed Central

    Tran, Thao T.; Dam, Phuongan; Su, Zhengchang; Poole, Farris L.; Adams, Michael W. W.; Zhou, G. Tong; Xu, Ying

    2007-01-01

    Identification of operons in the hyperthermophilic archaeon Pyrococcus furiosus represents an important step to understanding the regulatory mechanisms that enable the organism to adapt and thrive in extreme environments. We have predicted operons in P.furiosus by combining the results from three existing algorithms using a neural network (NN). These algorithms use intergenic distances, phylogenetic profiles, functional categories and gene-order conservation in their operon prediction. Our method takes as inputs the confidence scores of the three programs, and outputs a prediction of whether adjacent genes on the same strand belong to the same operon. In addition, we have applied Gene Ontology (GO) and KEGG pathway information to improve the accuracy of our algorithm. The parameters of this NN predictor are trained on a subset of all experimentally verified operon gene pairs of Bacillus subtilis. It subsequently achieved 86.5% prediction accuracy when applied to a subset of gene pairs for Escherichia coli, which is substantially better than any of the three prediction programs. Using this new algorithm, we predicted 470 operons in the P.furiosus genome. Of these, 349 were validated using DNA microarray data. PMID:17148478

  2. Pyrococcus furiosus strains and methods of using same

    SciTech Connect

    Lipscomb, Gina L; Farkas, Joel Andrew; Adams, Michael W. W.; Westpheling, Janet

    2015-01-06

    Provided herein are methods for transforming a Pyrococcus furiosus with a polynucleotide. In one embodiment, the method includes contacting a P. furiosus with a polynucleotide under conditions suitable for uptake of the polynucleotide by the P. furiosus, and identifying transformants at a frequency of, for instance, at least 10.sup.3 transformants per microgram DNA. Also provided are isolated Pyrococcus furiosus having the characteristics of Pyrococcus furiosus COM1, and plasmids that include an origin of replication that functions in a Pyrococcus furiosus. The plasmid is stable in a recipient P. furiosus without selection for more than 100 generations and is structurally unchanged after replication in P. furiosus for more than 100 generations.

  3. Characterization of a tungsten-iron-sulfur protein exhibiting novel spectroscopic and redox properties from the hyperthermophilic archaebacterium Pyrococcus furiosus.

    PubMed

    Mukund, S; Adams, M W

    1990-07-15

    The archaebacterium, Pyrococcus furiosus, is a strict anaerobe that grows optimally at 100 degrees C by a fermentative-type metabolism in which H2 and CO2 are the only detectable products. Tungsten is known to stimulate the growth of this organism. A red-colored tungsten-containing protein (abbreviated RTP) that is redox-active and extremely thermostable has been purified. RTP is a monomer of Mr = 85,000 and contains approximately 6 iron, 1 tungsten, and 4 acid-labile sulfide atoms/molecule. Titrations using visible spectroscopy were consistent with the oxidation and reduction of the protein each requiring two electrons/molecule, suggesting that these metals and the sulfide are arranged in two redox active centers. P. furiosus ferredoxin served as an electron acceptor for the protein. Dithionite-reduced RTP exhibited a remarkable and complex EPR spectrum at 6 K with g values ranging from 1.3 to 10.0. This was shown to arise from the spin-coupling interaction of two paramagnetic centers. One (center A) has a S = 3/2 spin system (effective g values: gx = 3.33, gy = 4.75, and gz = 1.92, where D = 4.3 cm-1 and lambda = 0.135), whereas the EPR properties of the other (center B) could not be deduced. Nevertheless, theoretical analyses show how the redox properties of both centers may be determined using EPR spectroscopy. Their midpoint potentials (Em) at 20 degrees C and pH 8.0 are -410 mV (center A) and -500 mV (center B) with an effective potential for the spin coupled system (Em, A + B) of -505 mV. The Em values are dependent on temperature (delta Em/delta T = -2 mV/degrees C between 20 and 70 degrees C) and pH with pK alpha values of 8.0 (A) and approximately 8.5 (B). The Em values at 100 degrees C, the growth temperature, were estimated at -590, -650, and -660 mV for centers A, B, and A + B, respectively. These data indicate that RTP catalyzes a dehydrogenase-type reaction of extremely low potential, which involves the transfer of two protons and of two electrons

  4. TrmB, a sugar sensing regulator of ABC transporter genes in Pyrococcus furiosus exhibits dual promoter specificity and is controlled by different inducers.

    PubMed

    Lee, Sung-Jae; Moulakakis, Christina; Koning, Sonja M; Hausner, Winfried; Thomm, Michael; Boos, Winfried

    2005-09-01

    TrmB is the transcriptional repressor for the gene cluster of the trehalose/maltose ABC transporter of the hyperthermophilic archaea Thermococcus litoralis and Pyrococcus furiosus (malE or TM operon), with maltose and trehalose acting as inducers. We found that TrmB (the protein is identical in both organisms) also regulated the transcription of genes encoding a separate maltodextrin ABC transporter in P. furiosus (mdxE or MD operon) with maltotriose, longer maltodextrins and sucrose acting as inducers, but not with maltose or trehalose. In vitro transcription of the malE and the mdxE operons was inhibited by TrmB binding to the different operator sequences. Inhibition of the TM operon was released by maltose and trehalose whereas inhibition of the MD operon was released by maltotriose and larger maltodextrins as well as by sucrose. Scanning mutagenesis of the TM operator revealed the role of the palindromic TACTNNNAGTA sequence for TrmB recognition. TrmB exhibits a broad spectrum of sugar-binding specificity, binding maltose, sucrose, maltotriose and trehalose in decreasing order of affinity, half-maximal binding occurring at 20, 60, 250 and 500 microM substrate concentration respectively. Of all substrates, only maltose shows sigmoidal binding characteristics with a Hill coefficient of 2. As measured by molecular sieve chromatography and cross-linking TrmB behaved as dimer in dilute buffer solution at room temperature. We conclude that TrmB acts as a bifunctional transcriptional regulator acting on two different promoters and being differentially controlled by binding to different sugars. We believe this to represent a novel strategy of prokaryotic transcription regulation.

  5. Pyrococcus Furiosus Genome Supplementary Data from the Adams Laboratory at the University of Georgia

    DOE Data Explorer

    Adams, Michael W.W.; Weinberg, Michael V.; Schut, Gerrit J.; Brehm, Scott; Datta, Susmitta; Zhou, J.

    The research in the Adams Laboratory focuses on the physiology of hyperthermophilic organisms with an emphasis on metal-containing enzymes in the hyperthermophilic marine archaeon Pyrococcus furiosus. Three of the many articles from this University of Georgia lab have supplementary materials that are available on the Adams Lab website. All three sets of data are Open Reading Frames (ORFs) used for DNA microarray experiments and the changes in signal intensities. The full citations for the three articles are: 1) Weinberg, M. V., Schut, G. J., Brehm, S., Datta, S. and Adams, M. W. W. (2005) Cold shock of a hyperthermophilic archaeon: Pyrococcus furiosus exhibits multiple responses to a suboptimal growth temperature with a key role for membrane-bound glycoproteins. J Bacteriol. 187, 336-348; 2) Schut, G. J., Brehm, S. D., Datta, S. and Adams, M. W. W. (2003) "Whole genome DNA microarray analysis of a hyperthermophile and an archaeon: Pyrococcus furiosus grown on carbohydrates or peptides" J. Bacteriol. 185, 3935-3947; Schut, G. J., Zhou, J. and Adams, M. W. W. (2001) "DNA microarray analysis of the hyperthermophilic archaeon Pyrococcus furiosus evidence for a new type of sulfur-reducing enzyme" J. Bacteriol. 183, 7027-7036. Note that these articles are copyrighted by the Journal of Bacteriology.

  6. Structure of hyperthermophilic β-glucosidase from Pyrococcus furiosus

    PubMed Central

    Kado, Yuji; Inoue, Tsuyoshi; Ishikawa, Kazuhiko

    2011-01-01

    Three categories of cellulases, endoglucanases, cellobiohydrolases and β-glucosidases, are commonly used in the process of cellulose saccharification. In particular, the activity and characteristics of hyperthermophilic β-glucosidase make it promising in industrial applications of biomass. In this paper, the crystal structure of the hyperthermophilic β-glucosidase from Pyrococcus furiosus (BGLPf) was determined at 2.35 Å resolution in a new crystal form. The structure showed that there is one tetramer in the asymmetric unit and that the dimeric molecule exhibits a structure that is stable towards sodium dodecyl sulfate (SDS). The dimeric molecule migrated in reducing SDS polyacrylamide gel electrophoresis (SDS–PAGE) buffer even after boiling at 368 K. Energy calculations demonstrated that one of the two dimer interfaces acquired the largest solvation free energy. Structural comparison and sequence alignment with mesophilic β-glucosidase A from Clostridium cellulovorans (BGLACc) revealed that the elongation at the C-terminal end forms a hydrophobic patch at the dimer interface that might contribute to hyperthermostability. PMID:22139147

  7. Production and Application of a Soluble Hydrogenase from Pyrococcus furiosus

    PubMed Central

    Wu, Chang-Hao; McTernan, Patrick M.; Walter, Mary E.; Adams, Michael W. W.

    2015-01-01

    Hydrogen gas is a potential renewable alternative energy carrier that could be used in the future to help supplement humanity's growing energy needs. Unfortunately, current industrial methods for hydrogen production are expensive or environmentally unfriendly. In recent years research has focused on biological mechanisms for hydrogen production and specifically on hydrogenases, the enzyme responsible for catalyzing the reduction of protons to generate hydrogen. In particular, a better understanding of this enzyme might allow us to generate hydrogen that does not use expensive metals, such as platinum, as catalysts. The soluble hydrogenase I (SHI) from the hyperthermophile Pyrococcus furiosus, a member of the euryarchaeota, has been studied extensively and used in various biotechnological applications. This review summarizes the strategies used in engineering and characterizing three different forms of SHI and the properties of the recombinant enzymes. SHI has also been used in in vitro systems for hydrogen production and NADPH generation and these systems are also discussed. PMID:26543406

  8. Production and Application of a Soluble Hydrogenase from Pyrococcus furiosus.

    PubMed

    Wu, Chang-Hao; McTernan, Patrick M; Walter, Mary E; Adams, Michael W W

    2015-01-01

    Hydrogen gas is a potential renewable alternative energy carrier that could be used in the future to help supplement humanity's growing energy needs. Unfortunately, current industrial methods for hydrogen production are expensive or environmentally unfriendly. In recent years research has focused on biological mechanisms for hydrogen production and specifically on hydrogenases, the enzyme responsible for catalyzing the reduction of protons to generate hydrogen. In particular, a better understanding of this enzyme might allow us to generate hydrogen that does not use expensive metals, such as platinum, as catalysts. The soluble hydrogenase I (SHI) from the hyperthermophile Pyrococcus furiosus, a member of the euryarchaeota, has been studied extensively and used in various biotechnological applications. This review summarizes the strategies used in engineering and characterizing three different forms of SHI and the properties of the recombinant enzymes. SHI has also been used in in vitro systems for hydrogen production and NADPH generation and these systems are also discussed. PMID:26543406

  9. Production and Application of a Soluble Hydrogenase from Pyrococcus furiosus

    DOE PAGESBeta

    Wu, Chang-Hao; McTernan, Patrick M.; Walter, Mary E.; Adams, Michael W. W.

    2015-01-01

    Hydrogen gas is a potential renewable alternative energy carrier that could be used in the future to help supplement humanity’s growing energy needs. Unfortunately, current industrial methods for hydrogen production are expensive or environmentally unfriendly. In recent years research has focused on biological mechanisms for hydrogen production and specifically on hydrogenases, the enzyme responsible for catalyzing the reduction of protons to generate hydrogen. In particular, a better understanding of this enzyme might allow us to generate hydrogen that does not use expensive metals, such as platinum, as catalysts. The soluble hydrogenase I (SHI) from the hyperthermophile Pyrococcus furiosus ,more » a member of the euryarchaeota, has been studied extensively and used in various biotechnological applications. This review summarizes the strategies used in engineering and characterizing three different forms of SHI and the properties of the recombinant enzymes. SHI has also been used in in vitro systems for hydrogen production and NADPH generation and these systems are also discussed.« less

  10. Characterization of amylolytic enzyme activities associated with the hyperthermophilic archaebacterium Pyrococcus furiosus

    SciTech Connect

    Brown, S.H.; Costantino, H.R.; Kelly, R.M. Univ. of Maryland, Baltimore )

    1990-07-01

    The hyperthermophilic archaebacterium Pyrococcus furiosus produces several amylolytic enzymes in response to the presence of complex carbohydrates in the growth medium. These enzyme activities, {alpha}-glucosidase, pullulanase, and {alpha}-amylase, were detected in both cell extracts and culture supernatants. All activities were characterized by temperature optima of at least 100{degree}C as well as a high degree of thermostability. The existence of this collection of activities in P. furiosus suggests that polysaccharide availability in its growth environment is a significant aspect of the niche from which it was isolated.

  11. Identification of two [4Fe-4S]-cluster-containing hydro-lyases from Pyrococcus furiosus.

    PubMed

    van Vugt-Lussenburg, Barbara M A; van der Weel, Laura; Hagen, Wilfred R; Hagedoorn, Peter-Leon

    2009-09-01

    The hyperthermophilic archaeon Pyrococcus furiosus is a strict anaerobe. It is therefore not expected to use the oxidative tricarboxylic acid (TCA) cycle for energy transduction. Nonetheless, its genome encodes more putative TCA cycle enzymes than the closely related Pyrococcus horikoshii and Pyrococcus abyssi, including an aconitase (PF0201). Furthermore, a two-subunit fumarase (PF1755 and PF1754) is encoded on the Pyr. furiosus genome. In the present study, these three genes were heterologously overexpressed in Escherichia coli to enable characterization of the enzymes. PF1755 and PF1754 were shown to form a [4Fe-4S]-cluster-containing heterodimeric enzyme, able to catalyse the reversible hydratation of fumarate. The aconitase PF0201 also contained an Fe-S cluster, and catalysed the conversion from citrate to isocitrate. The fumarase belongs to the class of two-subunit, [4Fe-4S]-cluster-containing fumarate hydratases exemplified by MmcBC from Pelotomaculum thermopropionicum; the aconitase belongs to the aconitase A family. Aconitase probably plays a role in amino acid synthesis when the organism grows on carbohydrates. However, the function of the seemingly metabolically isolated fumarase in Pyr. furiosus has yet to be established.

  12. A thermostable hybrid cluster protein from Pyrococcus furiosus: effects of the loss of a three helix bundle subdomain.

    PubMed

    Overeijnder, Marieke L; Hagen, Wilfred R; Hagedoorn, Peter-Leon

    2009-06-01

    Pyrococcus furiosus hybrid cluster protein (HCP) was expressed in Escherichia coli, purified, and characterized. This is the first archaeal and thermostable HCP to be isolated. Compared with the protein sequences of previously characterized HCPs from mesophiles, the protein sequence of P. furiosus HCP exhibits a deletion of approximately 13 kDa as a single amino acid stretch just after the N-terminal cysteine motif, characteristic for class-III HCPs from (hyper)thermophilic archaea and bacteria. The protein was expressed as a thermostable, soluble homodimeric protein. Hydroxylamine reductase activity of P. furiosus HCP showed a K(m) value of 0.40 mM and a k(cat) value of 3.8 s(-1) at 70 degrees C and pH 9.0. Electron paramagnetic resonance spectroscopy showed evidence for the presence of a spin-admixed, S = 3/2 [4Fe-4S](+) cubane cluster and of the hybrid cluster. The cubane cluster of P. furiosus HCP is presumably coordinated by a CXXC-X(7)-C-X(5)-C motif close to the N-terminus, which is similar to the CXXC-X(8)-C-X(5)-C motif of the Desulfovibrio desulfuricans and Desulfovibrio vulgaris HCPs. Amino acid sequence alignment and homology modeling of P. furiosus HCP reveal that the deletion results in a loss of one of the two three-helix bundles of domain 1. Clearly the loss of one of the three-helix bundles of domain 1 does not diminish the hydroxylamine reduction activity and the incorporation of the iron-sulfur clusters. PMID:19241093

  13. A thermostable hybrid cluster protein from Pyrococcus furiosus: effects of the loss of a three helix bundle subdomain.

    PubMed

    Overeijnder, Marieke L; Hagen, Wilfred R; Hagedoorn, Peter-Leon

    2009-06-01

    Pyrococcus furiosus hybrid cluster protein (HCP) was expressed in Escherichia coli, purified, and characterized. This is the first archaeal and thermostable HCP to be isolated. Compared with the protein sequences of previously characterized HCPs from mesophiles, the protein sequence of P. furiosus HCP exhibits a deletion of approximately 13 kDa as a single amino acid stretch just after the N-terminal cysteine motif, characteristic for class-III HCPs from (hyper)thermophilic archaea and bacteria. The protein was expressed as a thermostable, soluble homodimeric protein. Hydroxylamine reductase activity of P. furiosus HCP showed a K(m) value of 0.40 mM and a k(cat) value of 3.8 s(-1) at 70 degrees C and pH 9.0. Electron paramagnetic resonance spectroscopy showed evidence for the presence of a spin-admixed, S = 3/2 [4Fe-4S](+) cubane cluster and of the hybrid cluster. The cubane cluster of P. furiosus HCP is presumably coordinated by a CXXC-X(7)-C-X(5)-C motif close to the N-terminus, which is similar to the CXXC-X(8)-C-X(5)-C motif of the Desulfovibrio desulfuricans and Desulfovibrio vulgaris HCPs. Amino acid sequence alignment and homology modeling of P. furiosus HCP reveal that the deletion results in a loss of one of the two three-helix bundles of domain 1. Clearly the loss of one of the three-helix bundles of domain 1 does not diminish the hydroxylamine reduction activity and the incorporation of the iron-sulfur clusters.

  14. Expression and in vitro assembly of recombinant glutamate dehydrogenase from the hyperthermophilic archaeon Pyrococcus furiosus.

    PubMed Central

    Diruggiero, J; Robb, F T

    1995-01-01

    The gdhA gene, encoding the hexameric glutamate dehydrogenase (GDH) from the hyperthermophilic archaeon Pyrococcus furiosus, was expressed in Escherichia coli by using the pET11-d system. The recombinant GDH was soluble and constituted 15% of the E. coli cell extract. The N-terminal amino acid sequence of the recombinant protein was identical to the sequence of the P. furiosus enzyme, except for the presence of an initial methionine which was absent from the enzyme purified from P. furiosus. By molecular exclusion chromatography we showed that the recombinant GDH was composed of equal amounts of monomeric and hexameric forms. Heat treatment of the recombinant protein triggered in vitro assembly of inactive monomers into hexamers, resulting in increased GDH activity. The specific activity of the recombinant enzyme, purified by heat treatment and affinity chromatography, was equivalent to that of the native enzyme from P. furiosus. The recombinant GDH displayed a slightly lower level of thermostability, with a half-life of 8 h at 100 degrees C, compared with 10.5 h for the enzyme purified from P. furiosus. PMID:7887598

  15. Characterization of the glycolytic enzyme enolase which is abundant in the hyperthermophilic archaeon, Pyrococcus furiosus

    SciTech Connect

    Peak, M.J.; Peak, J.G.; Stevens, F.J.; Blamey, J.; Mai, X.; Zhou, Z.H.; Adams, M.W.W.

    1993-12-31

    High enolase activity, as measured by the conversion of 2-phosphoglycerate to phosphoenolphyruvate, was found in the cytoplasm of Pyrococcus (an anaerobic, hyperthermophilic archaeon that grows optimally at 100{degree}C). In this organism, the enzyme probably functions in a sugar fermentation pathway. The enzyme was purified to homogeneity. It had a temperature optimum of >90 {degree}C, and a pH optimum of 8.1. The enzyme was extremely thermostable with a half time for inactivation at 100{degree}C of 40 min. In contrast, an enolase from yeast was inactivated in 1 min at 88{degree}C. Both the P. furiosus and yeast enzymes required a metal ion for activity, but whereas the yeast enzyme has an absolute requirement for Mg{sup ++} the P. furiosus enolase was equally active in the presence of Mn{sup ++}. Both enzymes were competitively inhibited by citrate. P. furiosus enolase, as for mesophilic enolases, probably has a homodimeric structure with subunit M{sub r} greater than 45,000. A highly conserved sequence of eight amino acids in the N-terminal region was found in enolases from P. furiosus and a wide range of other organisms including bacteria, yeast, birds, and mammals.

  16. Characterization of UDP amino sugars as major phosphocompounds in the hyperthermophilic archaeon Pyrococcus furiosus.

    PubMed Central

    Ramakrishnan, V; Teng, Q; Adams, M W

    1997-01-01

    The archaeon Pyrococcus furiosus is a strictly anaerobic heterotroph that grows optimally at 100 degrees C by the fermentation of carbohydrates. It is known to contain high concentrations of novel intracellular solutes such as beta-mannosylglycerate and di-myo-inositol 1,1'-phosphate (DIP) (L. O. Martins and H. Santos, Appl. Environ. Microbiol. 61:3299-3303, 1995). Here, 31P nuclear magnetic resonance (NMR) spectroscopy was used to show that this organism also accumulates another type of phospho compound, as revealed by a major multiplet signal in the pyrophosphate region. The compounds were purified from cell extracts of P. furiosus by anion-exchange and gel filtration chromatographic procedures and were structurally analyzed by 1H, 13C, and 31P NMR spectroscopy. They were identified as two uridylated amino sugars, UDP N-acetylglucosamine and UDP N-acetylgalactosamine. Unambiguous characterizations and complete assignments of 1H and 13C resonances from such sugars have not been previously reported. In vitro 31P NMR spectroscopic analyses showed that, in contrast to DIP, which is maintained at a constant intracellular concentration (approximately 32 mM) throughout the growth phase of P. furiosus, the UDP amino sugars accumulated (to approximately 14 mM) only during the late log phase. The possible biochemical roles of these compounds in P. furiosus are discussed. PMID:9045806

  17. Novel and remarkably thermostable ferredoxin from the hyperthermophilic archaebacterium Pyrococcus furiosus

    SciTech Connect

    Aono, S.; Bryant, F.O.; Adams, M.W.W. )

    1989-06-01

    The archaebacterium Pyrococcus furiosus is a strict anaerobe that grows optimally at 100{degree}C by a fermentative-type metabolism in which H{sub 2} and CO{sub 2} are the only detectable products. A ferredoxin, which functions as the electron donor to the hydrogenase of this organism was purified under anaerobic reducing conditions. The study shows that P. furiosus ferredoxin is therefore distinct from other ferredoxins in that the bulk of its iron is not present as iron-sulfur clusters with an S = 1/2 ground state. The apoferredoxin was reconstituted with iron and sulfide to give a protein that was indistinguishable from the native ferredoxin by its iron content and electron paramagnetic resonance properties, which showed that the novel iron-sulfur clusters were not artifacts of purification. The reduced ferredoxin also functioned as an electron donor for H{sub 2} evolution catalyzed by the hydrogenase of the mesophilic eubacterium Clostridium pasteurianum. P. furiosus ferredoxin was resistant to denaturation by sodium dodecyl sulfate (20%, wt/vol) and was remarkably thermostable. Its UV-visible absorption spectrum and electron carrier activity to P. furiosus hydrogenase were unaffected by a 12-h incubation at 95{degree}C.

  18. Heterologous Production of an Energy-Conserving Carbon Monoxide Dehydrogenase Complex in the Hyperthermophile Pyrococcus furiosus

    PubMed Central

    Schut, Gerrit J.; Lipscomb, Gina L.; Nguyen, Diep M. N.; Kelly, Robert M.; Adams, Michael W. W.

    2016-01-01

    Carbon monoxide (CO) is an important intermediate in anaerobic carbon fixation pathways in acetogenesis and methanogenesis. In addition, some anaerobes can utilize CO as an energy source. In the hyperthermophilic archaeon Thermococcus onnurineus, which grows optimally at 80°C, CO oxidation and energy conservation is accomplished by a respiratory complex encoded by a 16-gene cluster containing a CO dehydrogenase, a membrane-bound [NiFe]-hydrogenase and a Na+/H+ antiporter module. This complex oxidizes CO, evolves CO2 and H2, and generates a Na+ motive force that is used to conserve energy by a Na+-dependent ATP synthase. Herein we used a bacterial artificial chromosome to insert the 13.2 kb gene cluster encoding the CO-oxidizing respiratory complex of T. onnurineus into the genome of the heterotrophic archaeon, Pyrococcus furiosus, which grows optimally at 100°C. P. furiosus is normally unable to utilize CO, however, the recombinant strain readily oxidized CO and generated H2 at 80°C. Moreover, CO also served as an energy source and allowed the P. furiosus strain to grow with a limiting concentration of sugar or with peptides as the carbon source. Moreover, CO oxidation by P. furiosus was also coupled to the re-utilization, presumably for biosynthesis, of acetate generated by fermentation. The functional transfer of CO utilization between Thermococcus and Pyrococcus species demonstrated herein is representative of the horizontal gene transfer of an environmentally relevant metabolic capability. The transfer of CO utilizing, hydrogen-producing genetic modules also has applications for biohydrogen production and a CO-based industrial platform for various thermophilic organisms. PMID:26858706

  19. Heterologous Production of an Energy-Conserving Carbon Monoxide Dehydrogenase Complex in the Hyperthermophile Pyrococcus furiosus.

    PubMed

    Schut, Gerrit J; Lipscomb, Gina L; Nguyen, Diep M N; Kelly, Robert M; Adams, Michael W W

    2016-01-01

    Carbon monoxide (CO) is an important intermediate in anaerobic carbon fixation pathways in acetogenesis and methanogenesis. In addition, some anaerobes can utilize CO as an energy source. In the hyperthermophilic archaeon Thermococcus onnurineus, which grows optimally at 80°C, CO oxidation and energy conservation is accomplished by a respiratory complex encoded by a 16-gene cluster containing a CO dehydrogenase, a membrane-bound [NiFe]-hydrogenase and a Na(+)/H(+) antiporter module. This complex oxidizes CO, evolves CO2 and H2, and generates a Na(+) motive force that is used to conserve energy by a Na(+)-dependent ATP synthase. Herein we used a bacterial artificial chromosome to insert the 13.2 kb gene cluster encoding the CO-oxidizing respiratory complex of T. onnurineus into the genome of the heterotrophic archaeon, Pyrococcus furiosus, which grows optimally at 100°C. P. furiosus is normally unable to utilize CO, however, the recombinant strain readily oxidized CO and generated H2 at 80°C. Moreover, CO also served as an energy source and allowed the P. furiosus strain to grow with a limiting concentration of sugar or with peptides as the carbon source. Moreover, CO oxidation by P. furiosus was also coupled to the re-utilization, presumably for biosynthesis, of acetate generated by fermentation. The functional transfer of CO utilization between Thermococcus and Pyrococcus species demonstrated herein is representative of the horizontal gene transfer of an environmentally relevant metabolic capability. The transfer of CO utilizing, hydrogen-producing genetic modules also has applications for biohydrogen production and a CO-based industrial platform for various thermophilic organisms. PMID:26858706

  20. Heterologous production of an energy-conserving carbon monoxide dehydrogenase complex in the hyperthermophile Pyrococcus furiosus

    DOE PAGESBeta

    Schut, Gerrit J.; Lipscomb, Gina L.; Nguyen, Diep M. N.; Kelly, Robert M.; Adams, Michael W. W.

    2016-01-29

    In this study, carbon monoxide (CO) is an important intermediate in anaerobic carbon fixation pathways in acetogenesis and methanogenesis. In addition, some anaerobes can utilize CO as an energy source. In the hyperthermophilic archaeon Thermococcus onnurineus, which grows optimally at 80°C, CO oxidation and energy conservation is accomplished by a respiratory complex encoded by a 16-gene cluster containing a CO dehydrogenase, a membrane-bound [NiFe]-hydrogenase and a Na+/H+ antiporter module. This complex oxidizes CO, evolves CO2 and H2, and generates a Na+ motive force that is used to conserve energy by a Na+-dependent ATP synthase. Herein we used a bacterial artificialmore » chromosome to insert the 13.2 kb gene cluster encoding the CO-oxidizing respiratory complex of T. onnurineus into the genome of the heterotrophic archaeon, Pyrococcus furiosus, which grows optimally at 100° C. P. furiosus is normally unable to utilize CO, however, the recombinant strain readily oxidized CO and generated H2 at 80° C. Moreover, CO also served as an energy source and allowed the P. furiosus strain to grow with a limiting concentration of sugar or with peptides as the carbon source. Moreover, CO oxidation by P. furiosus was also coupled to the re-utilization, presumably for biosynthesis, of acetate generated by fermentation. The functional transfer of CO utilization between Thermococcus and Pyrococcus species demonstrated herein is representative of the horizontal gene transfer of an environmentally relevant metabolic capability. The transfer of CO utilizing, hydrogen-producing genetic modules also has applications for biohydrogen production and a CO-based industrial platform for various thermophilic organisms.« less

  1. Phosphate and arsenate removal efficiency by thermostable ferritin enzyme from Pyrococcus furiosus using radioisotopes.

    PubMed

    Sevcenco, Ana-Maria; Paravidino, Monica; Vrouwenvelder, Johannes S; Wolterbeek, Hubert Th; van Loosdrecht, Mark C M; Hagen, Wilfred R

    2015-06-01

    Oxo-anion binding properties of the thermostable enzyme ferritin from Pyrococcus furiosus were characterized with radiography. Radioisotopes (32)P and (76)As present as oxoanions were used to measure the extent and the rate of their absorption by the ferritin. Thermostable ferritin proved to be an excellent system for rapid phosphate and arsenate removal from aqueous solutions down to residual concentrations at the picomolar level. These very low concentrations make thermostable ferritin a potential tool to considerably mitigate industrial biofouling by phosphate limitation or to remove arsenate from drinking water.

  2. The RNA- and DNA-targeting CRISPR–Cas immune systems of Pyrococcus furiosus

    PubMed Central

    Terns, Rebecca M.; Terns, Michael P.

    2014-01-01

    Using the hyperthermophile Pyrococcus furiosus, we have delineated several key steps in CRISPR (clustered regularly interspaced short palindromic repeats)–Cas (CRISPR-associated) invader defence pathways. P. furiosus has seven transcriptionally active CRISPR loci that together encode a total of 200 crRNAs (CRISPR RNAs). The 27 Cas proteins in this organism represent three distinct pathways and are primarily encoded in two large gene clusters. The Cas6 protein dices CRISPR locus transcripts to generate individual invader-targeting crRNAs. The mature crRNAs include a signature sequence element (the 5′ tag) derived from the CRISPR locus repeat sequence that is important for function. crRNAs are tailored into distinct species and integrated into three distinct crRNA–Cas protein complexes that are all candidate effector complexes. The complex formed by the Cmr [Cas module RAMP (repeat-associated mysterious proteins)] (subtype III-B) proteins cleaves complementary target RNAs and can be programmed to cleave novel target RNAs in a prokaryotic RNAi-like manner. Evidence suggests that the other two CRISPR–Cas systems in P. furiosus, Csa (Cas subtype Apern) (subtype I-A) and Cst (Cas subtype Tneap) (subtype I-B), target invaders at the DNA level. Studies of the CRISPR–Cas systems from P. furiosus are yielding fundamental knowledge of mechanisms of crRNA biogenesis and silencing for three of the diverse CRISPR–Cas pathways, and reveal that organisms such as P. furiosus possess an arsenal of multiple RNA-guided mechanisms to resist diverse invaders. Our knowledge of the fascinating CRISPR–Cas pathways is leading in turn to our ability to co-opt these systems for exciting new biomedical and biotechnological applications. PMID:24256230

  3. Structures of the superoxide reductase from Pyrococcus furiosus in the oxidized and reduced states.

    PubMed

    Yeh, A P; Hu, Y; Jenney, F E; Adams, M W; Rees, D C

    2000-03-14

    Superoxide reductase (SOR) is a blue non-heme iron protein that functions in anaerobic microbes as a defense mechanism against reactive oxygen species by catalyzing the reduction of superoxide to hydrogen peroxide [Jenney, F. E., Jr., Verhagen, M. F. J. M., Cui, X. , and Adams, M. W. W. (1999) Science 286, 306-309]. Crystal structures of SOR from the hyperthermophilic archaeon Pyrococcus furiosus have been determined in the oxidized and reduced forms to resolutions of 1.7 and 2.0 A, respectively. SOR forms a homotetramer, with each subunit adopting an immunoglobulin-like beta-barrel fold that coordinates a mononuclear, non-heme iron center. The protein fold and metal center are similar to those observed previously for the homologous protein desulfoferrodoxin from Desulfovibrio desulfuricans [Coelho, A. V., Matias, P., Fülöp, V., Thompson, A., Gonzalez, A., and Carrondo, M. A. (1997) J. Bioinorg. Chem. 2, 680-689]. Each iron is coordinated to imidazole nitrogens of four histidines in a planar arrangement, with a cysteine ligand occupying an axial position normal to this plane. In two of the subunits of the oxidized structure, a glutamate carboxylate serves as the sixth ligand to form an overall six-coordinate, octahedral coordinate environment. In the remaining two subunits, the sixth coordination site is either vacant or occupied by solvent molecules. The iron centers in all four subunits of the reduced structure exhibit pentacoordination. The structures of the oxidized and reduced forms of SOR suggest a mechanism by which superoxide accessibility may be controlled and define a possible binding site for rubredoxin, the likely physiological electron donor to SOR. PMID:10704199

  4. Expression of Pyrococcus furiosus Superoxide Reductase in Arabidopsis Enhances Heat Tolerance1[C][W][OA

    PubMed Central

    Im, Yang Ju; Ji, Mikyoung; Lee, Alice; Killens, Rushyannah; Grunden, Amy M.; Boss, Wendy F.

    2009-01-01

    Plants produce reactive oxygen species (ROS) in response to environmental stresses sending signaling cues, which, if uncontrolled, result in cell death. Like other aerobic organisms, plants have ROS-scavenging enzymes, such as superoxide dismutase (SOD), which removes superoxide anion radical (O2−) and prevents the production and buildup of toxic free radicals. However, increasing the expression of cytosolic SODs is complex, and increasing their production in vivo has proven to be challenging. To avoid problems with endogenous regulation of gene expression, we expressed a gene from the archaeal hyperthermophile Pyrococcus furiosus that reduces O2−. P. furiosus uses superoxide reductase (SOR) rather than SOD to remove superoxide. SOR is a thermostable enzyme that reduces O2− in a one-electron reduction without producing oxygen. We show that P. furiosus SOR can be produced as a functional enzyme in planta and that plants producing SOR have enhanced tolerance to heat, light, and chemically induced ROS. Stress tolerance in the SOR-producing plants correlates positively with a delayed increase in ROS-sensitive transcripts and a decrease in ascorbate peroxidase activity. The SOR plants provide a good model system to study the impact of cytosolic ROS on downstream signaling in plant growth and development. Furthermore, this work demonstrates that this synthetic approach for reducing cytosolic ROS holds promise as a means for improving stress tolerance in crop plants. PMID:19684226

  5. Engineering Hyperthermophilic Archaeon Pyrococcus furiosus to Overproduce Its Cytoplasmic [NiFe]-Hydrogenase*

    PubMed Central

    Chandrayan, Sanjeev K.; McTernan, Patrick M.; Hopkins, R. Christopher; Sun, Junsong; Jenney, Francis E.; Adams, Michael W. W.

    2012-01-01

    The cytoplasmic hydrogenase (SHI) of the hyperthermophilic archaeon Pyrococcus furiosus is an NADP(H)-dependent heterotetrameric enzyme that contains a nickel-iron catalytic site, flavin, and six iron-sulfur clusters. It has potential utility in a range of bioenergy systems in vitro, but a major obstacle in its use is generating sufficient amounts. We have engineered P. furiosus to overproduce SHI utilizing a recently developed genetic system. In the overexpression (OE-SHI) strain, transcription of the four-gene SHI operon was under the control of a strong constitutive promoter, and a Strep-tag II was added to the N terminus of one subunit. OE-SHI and wild-type P. furiosus strains had similar rates of growth and H2 production on maltose. Strain OE-SHI had a 20-fold higher transcription of the polycistronic hydrogenase mRNA encoding SHI, and the specific activity of the cytoplasmic hydrogenase was ∼10-fold higher when compared with the wild-type strain, although the expression levels of genes encoding processing and maturation of SHI were the same in both strains. Overexpressed SHI was purified by a single affinity chromatography step using the Strep-tag II, and it and the native form had comparable activities and physical properties. Based on protein yield per gram of cells (wet weight), the OE-SHI strain yields a 100-fold higher amount of hydrogenase when compared with the highest homologous [NiFe]-hydrogenase system previously reported (from Synechocystis). This new P. furiosus system will allow further engineering of SHI and provide hydrogenase for efficient in vitro biohydrogen production. PMID:22157005

  6. Studies on Hydrogen Production by Photosynthetic Bacteria after Anaerobic Fermentation of Starch by a Hyperthermophile, Pyrococcus furiosus

    NASA Astrophysics Data System (ADS)

    Sugitate, Toshihiro; Fukatsu, Makoto; Ishimi, Katsuhiro; Kohno, Hideki; Wakayama, Tatsuki; Nakamura, Yoshihiro; Miyake, Jun; Asada, Yasuo

    In order to establish the sequential hydrogen production from waste starch using a hyperthermophile, Pyrococcus furiosus, and a photosynthetic bacterium, basic studies were done. P. furiosus produced hydrogen and acetate by anaerobic fermentation at 90°C. A photosynthetic bacterium, Rhodobacter sphaeroides RV, was able to produce hydrogen from acetate under anaerobic and light conditions at 30°C. However, Rb. sphaeroides RV was not able to produce hydrogen from acetate in the presence of sodium chloride that was essential for the growth and hydrogen production of P. furiosus although it produced hydrogen from lactate at a reduced rate with 1% sodium chloride. A newly isolated strain, CST-8, from natural environment was, however, able to produce hydrogen from acetate, especially with 3 mM L-alanine and in the presence of 1% sodium chloride. The sequential hydrogen production with P. furiosus and salt-tolerant photosynthetic bacteria could be probable at least in the laboratory experiment scale.

  7. Recombinant expression library of Pyrococcus furiosus constructed by high-throughput cloning: a useful tool for functional and structural genomics

    PubMed Central

    Yuan, Hui; Peng, Li; Han, Zhong; Xie, Juan-Juan; Liu, Xi-Peng

    2015-01-01

    Hyperthermophile Pyrococcus furiosus grows optimally near 100°C and is an important resource of many industrial and molecular biological enzymes. To study the structure and function of P. furiosus proteins at whole genome level, we constructed expression plasmids of each P. furiosus gene using a ligase-independent cloning method, which was based on amplifying target gene and vector by PCR using phosphorothioate-modified primers and digesting PCR products by λ exonuclease. Our cloning method had a positive clone percentage of ≥ 80% in 96-well plate cloning format. Small-scale expression experiment showed that 55 out of 80 genes were efficiently expressed in Escherichia coli Strain Rosetta 2(DE3)pLysS. In summary, this recombinant expression library of P. furiosus provides a platform for functional and structural studies, as well as developing novel industrial enzymes. Our cloning scheme is adaptable to constructing recombinant expression library of other sequenced organisms. PMID:26441878

  8. Cloning, purification, crystallization and preliminary crystallographic analysis of a hypothetical acetyltransferase from Pyrococcus furiosus

    PubMed Central

    Biarrotte-Sorin, Sabrina; Mayer, Claudine

    2005-01-01

    The GCN5-related N-acetyltransferase (GNAT) superfamily has a primordial role in cellular processes such as transcription initiation and regulation by histone acetylation, aminoglycoside resistance and melatonin metabolism. To date, no acetyltransferase from the archaeal domain of life has been studied. This paper describes the cloning, expression, purification and crystallization of a Pyrococcus furiosus hypothetical acetyltransferase PfGNAT (MW = 22 007 Da). The crystals belong to space group P622, with one molecule in the asymmetric unit and unit-cell parameters a = b = 82.6, c = 105.92 Å, α = β = 90, γ = 120°. Crystals diffract X-rays to 3.0 Å resolution on a synchrotron-radiation source. Determination of this structure will provide new insights into the substrate-specificity of this acetyltransferase and the thermal stability of the N-acetyltransferase domain. PMID:16511014

  9. Comparative Analysis of Barophily-Related Amino Acid Content in Protein Domains of Pyrococcus abyssi and Pyrococcus furiosus

    PubMed Central

    Yafremava, Liudmila S.; Di Giulio, Massimo; Caetano-Anollés, Gustavo

    2013-01-01

    Amino acid substitution patterns between the nonbarophilic Pyrococcus furiosus and its barophilic relative P. abyssi confirm that hydrostatic pressure asymmetry indices reflect the extent to which amino acids are preferred by barophilic archaeal organisms. Substitution patterns in entire protein sequences, shared protein domains defined at fold superfamily level, domains in homologous sequence pairs, and domains of very ancient and very recent origin now provide further clues about the environment that led to the genetic code and diversified life. The pyrococcal proteomes are very similar and share a very early ancestor. Relative amino acid abundance analyses showed that biases in the use of amino acids are due to their shared fold superfamilies. Within these repertoires, only two of the five amino acids that are preferentially barophilic, aspartic acid and arginine, displayed this preference significantly and consistently across structure and in domains appearing in the ancestor. The more primordial asparagine, lysine and threonine displayed a consistent preference for nonbarophily across structure and in the ancestor. Since barophilic preferences are already evident in ancient domains that are at least ~3 billion year old, we conclude that barophily is a very ancient trait that unfolded concurrently with genetic idiosyncrasies in convergence towards a universal code. PMID:24187517

  10. SurR regulates hydrogen production in Pyrococcus furiosus by a sulfur-dependent redox switch

    PubMed Central

    Yang, Hua; Lipscomb, Gina L.; Keese, Annette M.; Schut, Gerrit J.; Thomm, Michael; Adams, Michael W. W.; Wang, Bi Cheng; Scott, Robert A.

    2010-01-01

    SUMMARY We present structural and biochemical evidence for a redox switch in the archaeal transcriptional regulator SurR of Pyrococcus furiosus, a hyperthermophilic anaerobe. P. furiosus produces H2 during fermentation, but undergoes a metabolic shift to produce H2S when elemental sulfur (S0) becomes available. Changes in gene expression occur within minutes of S0 addition, and the majority of these S0-responsive genes are regulatory targets of SurR, a key regulator involved in primary S0 response. SurR was shown in vitro to have dual functionality, activating transcription of some of these genes, notably the hydrogenase operons, and repressing others, including a gene encoding sulfur reductase. This work demonstrates via biochemical and structural evidence that the activity of SurR is modulated by cysteine residues in a CxxC motif that constitute a redox switch. Oxidation of the switch with S0 inhibits sequence-specific DNA binding by SurR, leading to deactivation of genes related to H2 production and derepression of genes involved in S0 metabolism. PMID:20598080

  11. Intact Functional Fourteen-subunit Respiratory Membrane-bound [NiFe]-Hydrogenase Complex of the Hyperthermophilic Archaeon Pyrococcus furiosus*

    PubMed Central

    McTernan, Patrick M.; Chandrayan, Sanjeev K.; Wu, Chang-Hao; Vaccaro, Brian J.; Lancaster, W. Andrew; Yang, Qingyuan; Fu, Dax; Hura, Greg L.; Tainer, John A.; Adams, Michael W. W.

    2014-01-01

    The archaeon Pyrococcus furiosus grows optimally at 100 °C by converting carbohydrates to acetate, CO2, and H2, obtaining energy from a respiratory membrane-bound hydrogenase (MBH). This conserves energy by coupling H2 production to oxidation of reduced ferredoxin with generation of a sodium ion gradient. MBH is encoded by a 14-gene operon with both hydrogenase and Na+/H+ antiporter modules. Herein a His-tagged MBH was expressed in P. furiosus and the detergent-solubilized complex purified under anaerobic conditions by affinity chromatography. Purified MBH contains all 14 subunits by electrophoretic analysis (13 subunits were also identified by mass spectrometry) and had a measured iron:nickel ratio of 15:1, resembling the predicted value of 13:1. The as-purified enzyme exhibited a rhombic EPR signal characteristic of the ready nickel-boron state. The purified and membrane-bound forms of MBH both preferentially evolved H2 with the physiological donor (reduced ferredoxin) as well as with standard dyes. The O2 sensitivities of the two forms were similar (half-lives of ∼15 h in air), but the purified enzyme was more thermolabile (half-lives at 90 °C of 1 and 25 h, respectively). Structural analysis of purified MBH by small angle x-ray scattering indicated a Z-shaped structure with a mass of 310 kDa, resembling the predicted value (298 kDa). The angle x-ray scattering analyses reinforce and extend the conserved sequence relationships of group 4 enzymes and complex I (NADH quinone oxidoreductase). This is the first report on the properties of a solubilized form of an intact respiratory MBH complex that is proposed to evolve H2 and pump Na+ ions. PMID:24860091

  12. Intact functional fourteen-subunit respiratory membrane-bound [NiFe]-hydrogenase complex of the hyperthermophilic archaeon Pyrococcus furiosus.

    PubMed

    McTernan, Patrick M; Chandrayan, Sanjeev K; Wu, Chang-Hao; Vaccaro, Brian J; Lancaster, W Andrew; Yang, Qingyuan; Fu, Dax; Hura, Greg L; Tainer, John A; Adams, Michael W W

    2014-07-11

    The archaeon Pyrococcus furiosus grows optimally at 100 °C by converting carbohydrates to acetate, CO2, and H2, obtaining energy from a respiratory membrane-bound hydrogenase (MBH). This conserves energy by coupling H2 production to oxidation of reduced ferredoxin with generation of a sodium ion gradient. MBH is encoded by a 14-gene operon with both hydrogenase and Na(+)/H(+) antiporter modules. Herein a His-tagged MBH was expressed in P. furiosus and the detergent-solubilized complex purified under anaerobic conditions by affinity chromatography. Purified MBH contains all 14 subunits by electrophoretic analysis (13 subunits were also identified by mass spectrometry) and had a measured iron:nickel ratio of 15:1, resembling the predicted value of 13:1. The as-purified enzyme exhibited a rhombic EPR signal characteristic of the ready nickel-boron state. The purified and membrane-bound forms of MBH both preferentially evolved H2 with the physiological donor (reduced ferredoxin) as well as with standard dyes. The O2 sensitivities of the two forms were similar (half-lives of ∼ 15 h in air), but the purified enzyme was more thermolabile (half-lives at 90 °C of 1 and 25 h, respectively). Structural analysis of purified MBH by small angle x-ray scattering indicated a Z-shaped structure with a mass of 310 kDa, resembling the predicted value (298 kDa). The angle x-ray scattering analyses reinforce and extend the conserved sequence relationships of group 4 enzymes and complex I (NADH quinone oxidoreductase). This is the first report on the properties of a solubilized form of an intact respiratory MBH complex that is proposed to evolve H2 and pump Na(+) ions. PMID:24860091

  13. Identification of membrane proteins in the hyperthermophilic archaeon Pyrococcus furiosus using proteomics and prediction programs.

    SciTech Connect

    Holden, J. F.; Poole, F. L.; Tollaksen, S. L.; Giometti, C. S.; Lim, H.; Yates, J. R.; Adams, M. W. W.; Biosciences Division; Univ. of Georgia; The Scnpps Research Inst.

    2001-01-01

    Cell-free extracts from the hyperthermophilic archaeon Pyrococcus furiosus were separated into membrane and cytoplasmic fractions and each was analyzed by 2D-gel electrophoresis. A total of 66 proteins were identified, 32 in the membrane fraction and 34 in the cytoplasmic fraction. Six prediction programs were used to predict the subcellular locations of these proteins. Three were based on signal-peptides (SignalP, TargetP, and SOSUISignal) and three on transmembrane-spanning a-helices (TSEG, SOSUI, and PRED-TMR2). A consensus of the six programs predicted that 23 of the 32 proteins (72%) from the membrane fraction should be in the membrane and that all of the proteins from the cytoplasmic fraction should be in the cytoplasm. Two membrane-associated proteins predicted to be cytoplasmic by the programs are also predicted to consist primarily of transmembrane-spanning {beta}-sheets using porin protein models, suggesting that they are, in fact, membrane components. An ATPase subunit homolog found in the membrane fraction, although predicted to be cytoplasmic, is most likely complexed with other ATPase subunits in the membrane fraction. An additional three proteins predicted to be cytoplasmic but found in the membrane fraction, may be cytoplasmic contaminants. These include a chaperone homolog that may have attached to denatured membrane proteins during cell fractionation. Omitting these three proteins would boost the membrane-protein predictability of the models to near 80%. A consensus prediction using all six programs for all 2242 ORFs in the P. furiosus genome estimates that 24% of the ORF products are found in the membrane. However, this is likely to be a minimum value due to the programs' inability to recognize certain membrane-related proteins, such as subunits associated with membrane complexes and porin-type proteins.

  14. Identification of Membrane Proteins in the Hyperthermophilic Archaeon Pyrococcus Furiosus Using Proteomics and Prediction Programs

    PubMed Central

    Holden, James F.; Poole II, Farris L.; Tollaksen, Sandra L.; Giometti, Carol S.; Lim, Hanjo; Yates III, John R.

    2001-01-01

    Cell-free extracts from the hyperthermophilic archaeon Pyrococcus furiosus were separated into membrane and cytoplasmic fractions and each was analyzed by 2D-gel electrophoresis. A total of 66 proteins were identified, 32 in the membrane fraction and 34 in the cytoplasmic fraction. Six prediction programs were used to predict the subcellular locations of these proteins. Three were based on signal-peptides (SignalP, TargetP, and SOSUISignal) and three on transmembrane-spanning α-helices (TSEG, SOSUI, and PRED-TMR2). A consensus of the six programs predicted that 23 of the 32 proteins (72%) from the membrane fraction should be in the membrane and that all of the proteins from the cytoplasmic fraction should be in the cytoplasm. Two membrane-associated proteins predicted to be cytoplasmic by the programs are also predicted to consist primarily of transmembrane-spanning β-sheets using porin protein models, suggesting that they are, in fact, membrane components. An ATPase subunit homolog found in the membrane fraction, although predicted to be cytoplasmic, is most likely complexed with other ATPase subunits in the membrane fraction. An additional three proteins predicted to be cytoplasmic but found in the membrane fraction, may be cytoplasmic contaminants. These include a chaperone homolog that may have attached to denatured membrane proteins during cell fractionation. Omitting these three proteins would boost the membrane-protein predictability of the models to near 80%. A consensus prediction using all six programs for all 2242 ORFs in the P. furiosus genome estimates that 24% of the ORF products are found in the membrane. However, this is likely to be a minimum value due to the programs’ inability to recognize certain membrane-related proteins, such as subunits associated with membrane complexes and porin-type proteins. PMID:18629240

  15. Differential signal transduction via TrmB, a sugar sensing transcriptional repressor of Pyrococcus furiosus.

    PubMed

    Lee, Sung-Jae; Surma, Melanie; Seitz, Sabine; Hausner, Winfried; Thomm, Michael; Boos, Winfried

    2007-06-01

    TrmB is a transcriptional repressor of the hyperthermophilic archaeon Pyrococcus furiosus serving at least two operons. TrmB represses genes encoding an ABC transporter for trehalose and maltose (the TM system) with trehalose and maltose as inducers. TrmB also represses genes encoding another ABC transporter for maltodextrins (the MD system) with maltotriose and sucrose as inducers. Here we report that glucose which was also bound by TrmB acted as a corepressor (causing stronger repression) for both the TM and the MD system. Binding of glucose by TrmB was increased in the presence of TM promoter DNA. Maltose which acted as inducer for the TM system acted as a corepressor for the MD system intensifying repression. We propose that the differential conformational changes of TrmB in response to binding the different sugars governs the ability of TrmB to interact with the promoter region and represents a simple mechanism for selecting the usage of one carbon source over the other, reminiscent of catabolite repression in bacteria.

  16. A new strategy to express the extracellular α-amylase from Pyrococcus furiosus in Bacillus amyloliquefaciens

    PubMed Central

    Wang, Ping; Wang, Peili; Tian, Jian; Yu, Xiaoxia; Chang, Meihui; Chu, Xiaoyu; Wu, Ningfeng

    2016-01-01

    Extracellular α-amylase from Pyrococcus furiosus (PFA) shows great starch-processing potential for industrial application due to its thermostability, long half-life and optimal activity at low pH; however, it is difficult to produce in large quantities. In contrast, α-amylase from Bacillus amyloliquefaciens (BAA) can be produced in larger quantities, but shows lower stability at high temperatures and low pH. Here, we describe a BAA protein expression pattern-mimicking strategy to express PFA in B. amyloliquefaciens using the expression and secretion elements of BAA, including the codon usage bias and mRNA structure of gene, promoter, signal peptide, host and cultivation conditions. This design was assessed to be successful by comparing the various genes (mpfa and opfa), promoters (PamyA and P43), and strains (F30, F31, F32 and F30-∆amyA). The final production of PFA yielded 2714 U/mL, about 3000- and 14-fold that reportedly produced in B. subtilis or E. coli, respectively. The recombinant PFA was optimally active at ~100 °C and pH 5 and did not require Ca2+ for activity or thermostability, and >80% of the enzyme activity was retained after treatment at 100 °C for 4 h. PMID:26916714

  17. A cryo-crystallographic time course for peroxide reduction by rubrerythrin from Pyrococcus furiosus

    SciTech Connect

    Dillard, Bret D.; Demick, Jonathan M.; Adams, Michael W.W.; Lanzilotta, William N.

    2011-09-06

    High-resolution crystal structures of Pyrococcus furiosus rubrerythrin (PfRbr) in the resting (all-ferrous) state and at time points following exposure of the crystals to hydrogen peroxide are reported. This approach was possible because of the relativity slow turnover of PfRbr at room temperature. To this end, we were able to perform time-dependent peroxide treatment of the fully reduced enzyme, under strictly anaerobic conditions, in the crystalline state. In this work we demonstrate, for the first time, that turnover of a thermophilic rubrerythrin results in approximately 2-{angstrom} movement of one iron atom in the diiron site from a histidine to a carboxylate ligand. These results confirm that, despite the domain-swapped architecture, the hyperthermophilic rubrerythrins also utilize the classic combination of iron sites together with redox-dependent iron toggling to selectively reduce hydrogen peroxide over dioxygen. In addition, we have identified previously unobserved intermediates in the reaction cycle and observed structural changes that may explain the enzyme precipitation observed for the all-iron form of PfRbr upon oxidation to the all-ferric state.

  18. Preliminary neutron crystallographic analysis of selectively CH3-protonated, deuterated rubredoxin from Pyrococcus furiosus

    SciTech Connect

    Weiss, Kevin L; Meilleur, Flora; Blakeley, Matthew; Myles, Dean A A

    2008-01-01

    Neutron crystallography is used to locate hydrogen atoms in biological materials and can distinguish between negatively scattering hydrogen and positively scattering deuterium substituted positions in isomorphous neutron structures. Recently, Hauptman and Langs (2003) have shown that neutron diffraction data can be used to solve macromolecular structures by direct methods and that solution is aided by the presence of negatively scattering hydrogen atoms in the structure. Selective labeling protocols allow the design and production of H/D-labeled macromolecular structures in which the ratio of hydrogen to deuterium atoms can be precisely controlled. We have applied methyl-selective labeling protocols to introduce (1H-delta methyl)-leucine and (1H-gamma methyl)-valine into deuterated rubredoxin from Pyrococcus furiosus (PfRd). Here we report on the production, crystallization, and preliminary neutron analysis of the selectively CH3-protonated, deuterated PfRd sample, which provided a high quality neutron data set extending to 1.75 resolution at the new LADI-III instrument at the Insititut Laue-Langevin. Preliminary analysis of neutron density maps allows unambiguous assignment of the positions of hydrogen atoms at the methyl groups of the valine and leucine residues in the otherwise deuterated rubredoxin structure.

  19. Cloning, expression, and molecular characterization of the gene encoding an extremely thermostable [4Fe-4S] ferredoxin from the hyperthermophilic archaeon Pyrococcus furiosus.

    PubMed Central

    Heltzel, A; Smith, E T; Zhou, Z H; Blamey, J M; Adams, M W

    1994-01-01

    The gene for ferredoxin from the hyperthermophilic archaeon Pyrococcus furiosus was cloned, sequenced, and expressed in Escherichia coli. The coding region confirmed the determined amino acid sequence. Putative archaeon-type transcriptional regulatory elements were identified. The fdxA gene appears to be an independent transcriptional unit. Recombinant ferredoxin was indistinguishable from the protein purified from P. furiosus in its thermal stability and in the potentiometric and spectroscopic properties of its [4Fe-4S] cluster. PMID:8045914

  20. Cloning, sequencing, and expression of the gene encoding amylopullulanase from Pyrococcus furiosus and biochemical characterization of the recombinant enzyme.

    PubMed

    Dong, G; Vieille, C; Zeikus, J G

    1997-09-01

    The gene encoding the Pyrococcus furiosus hyperthermophilic amylopullulanase (APU) was cloned, sequenced, and expressed in Escherichia coli. The gene encoded a single 827-residue polypeptide with a 26-residue signal peptide. The protein sequence had very low homology (17 to 21% identity) with other APUs and enzymes of the alpha-amylase family. In particular, none of the consensus regions present in the alpha-amylase family could be identified. P. furiosus APU showed similarity to three proteins, including the P. furiosus intracellular alpha-amylase and Dictyoglomus thermophilum alpha-amylase A. The mature protein had a molecular weight of 89,000. The recombinant P. furiosus APU remained folded after denaturation at temperatures of < or = 70 degrees C and showed an apparent molecular weight of 50,000 in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Denaturating temperatures of above 100 degrees C were required for complete unfolding. The enzyme was extremely thermostable, with an optimal activity at 105 degrees C and pH 5.5. Ca2+ increased the enzyme activity, thermostability, and substrate affinity. The enzyme was highly resistant to chemical denaturing reagents, and its activity increased up to twofold in the presence of surfactants.

  1. Mechanism of oxygen detoxification by the surprisingly oxygen-tolerant hyperthermophilic archaeon, Pyrococcus furiosus.

    PubMed

    Thorgersen, Michael P; Stirrett, Karen; Scott, Robert A; Adams, Michael W W

    2012-11-01

    The anaerobic archaeon Pyrococcus furiosus grows by fermenting carbohydrates producing H(2), CO(2), and acetate. We show here that it is surprisingly tolerant to oxygen, growing well in the presence of 8% (vol/vol) O(2). Although cell growth and acetate production were not significantly affected by O(2), H(2) production was reduced by 50% (using 8% O(2)). The amount of H(2) produced decreased in a linear manner with increasing concentrations of O(2) over the range 2-12% (vol/vol), and for each mole of O(2) consumed, the amount of H(2) produced decreased by approximately 2 mol. The recycling of H(2) by the two cytoplasmic hydrogenases appeared not to play a role in O(2) resistance because a mutant strain lacking both enzymes was not more sensitive to O(2) than the parent strain. Decreased H(2) production was also not due to inactivation of the H(2)-producing, ferredoxin-dependent membrane-bound hydrogenase because its activity was unaffected by O(2) exposure. Electrons from carbohydrate oxidation must therefore be diverted to relieve O(2) stress at the level of reduced ferredoxin before H(2) production. Deletion strains lacking superoxide reductase (SOR) and putative flavodiiron protein A showed increased sensitivity to O(2), indicating that these enzymes play primary roles in resisting O(2). However, a mutant strain lacking the proposed electron donor to SOR, rubredoxin, was unaffected in response to O(2). Hence, electrons from sugar oxidation normally used to produce H(2) are diverted to O(2) detoxification by SOR and putative flavodiiron protein A, but the electron flow pathway from ferredoxin does not necessarily involve rubredoxin.| PMID:23093671

  2. Recombinant production of hyperthermostable CelB from Pyrococcus furiosus in Lactobacillus sp.

    PubMed

    Böhmer, N; Lutz-Wahl, S; Fischer, L

    2012-11-01

    Lactic acid bacteria (LAB) are used widespread in the food industry as traditional starters for various fermented foods. For recombinant protein production, LAB would be superior with view from the food safety demands since most of them are Generally Recognized As Safe organisms. We investigated the two pSIP expression systems, pSIP403 and pSIP409 (Sørvig et al. 2005), to produce a hyper-thermophilic β-glycosidase (CelB) from Pyrococcus furiosus in Lactobacillus plantarum NC8 and Lactobacillus casei as hosts, respectively. Both lactobacilli harboring the pSIP409-celB vector produced active CelB in batch bioreactor cultivations (MRS medium) while the specific CelB activity of the cell free extract was about 44 % higher with L. plantarum (1,590 ± 90 nkat/mg(protein)) than with L. casei (1,070 ± 66 nkat/mg(protein)) using p-nitrophenyl-β-galactoside (pNPGal) as the substrate. A fed-batch bioreactor cultivation of L. plantarum NC8 pSIP409-celB resulted in a specific CelB activity of 2,500 ± 120 nkat ( pNPGal)/mg(protein) after 28 h. A repeated dosage of the inducer spp-IP did not increase the enzyme expression further. As alternative for the cost intensive MRS medium, a basal whey medium with supplements (yeast extract, Tween 80, NH(4)-citrate) was developed. In bioreactor cultivations using this medium, about 556 ± 29 nkat ( pNPGal)/mg(protein) of CelB activity was achieved. It was shown that both LAB were potential expression hosts for recombinant enzyme production. The pSIP expression system can be applied in L. casei.

  3. Inhibition and stimulation of formation of the ferroxidase center and the iron core in Pyrococcus furiosus ferritin.

    PubMed

    Honarmand Ebrahimi, Kourosh; Hagedoorn, Peter-Leon; Hagen, Wilfred R

    2010-11-01

    Ferritin is a ubiquitous iron-storage protein that has 24 subunits. Each subunit of ferritins that exhibit high Fe(II) oxidation rates has a diiron binding site, the so-called ferroxidase center (FC). The role of the FC appears to be essential for the iron-oxidation catalysis of ferritins. Studies of the iron oxidation by mammalian, bacterial, and archaeal ferritin have indicated different mechanisms are operative for Fe(II) oxidation, and for inhibition of the Fe(II) oxidation by Zn(II). These differences are presumably related to the variations in the amino acid residues of the FC and/or transport channels. We have used a combination of UV-vis spectroscopy, fluorescence spectroscopy, and isothermal titration calorimetry to study the inhibiting action of Zn(II) ions on the iron-oxidation process by apoferritin and by ferritin aerobically preloaded with 48 Fe(II) per 24-meric protein, and to study a possible role of phosphate in initial iron mineralization by Pyrococcus furiosus ferritin (PfFtn). Although the empty FC can accommodate two zinc ions, binding of one zinc ion to the FC suffices to essentially abolish iron-oxidation activity. Zn(II) no longer binds to the FC nor does it inhibit iron core formation once the FC is filled with two Fe(III). Phosphate and vanadate facilitate iron oxidation only after formation of a stable FC, whereupon they become an integral part of the core. These results corroborate our previous proposal that the FC in PfFtn is a stable prosthetic group, and they suggest that its formation is essential for iron-oxidation catalysis by the protein.

  4. Characterization of Ten Heterotetrameric NDP-Dependent Acyl-CoA Synthetases of the Hyperthermophilic Archaeon Pyrococcus furiosus

    DOE PAGESBeta

    Scott, Joseph W.; Poole, Farris L.; Adams, Michael W. W.

    2014-01-01

    Tmore » he hyperthermophilic archaeon Pyrococcus furiosus grows by fermenting peptides and carbohydrates to organic acids. In the terminal step, acyl-CoA synthetase (ACS) isoenzymes convert acyl-CoA derivatives to the corresponding acid and conserve energy in the form of ATP. ACS1 and ACS2 were previously purified from P. furiosus and have α 2 β 2 structures but the genome contains genes encoding three additional α -subunits.he ten possible combinations of α and β genes were expressed in E. coli and each resulted in stable and active α 2 β 2 isoenzymes.he α -subunit of each isoenzyme determined CoA-based substrate specificity and between them they accounted for the CoA derivatives of fourteen amino acids.he β -subunit determined preference for adenine or guanine nucleotides.he GTP-generating isoenzymes are proposed to play a role in gluconeogenesis by producing GTP for GTP-dependent phosphoenolpyruvate carboxykinase and for other GTP-dependent processes.ranscriptional and proteomic data showed that all ten isoenzymes are constitutively expressed indicating that both ATP and GTP are generated from the metabolism of most of the amino acids. A phylogenetic analysis showed that the ACSs of P. furiosus and other members of thehermococcales are evolutionarily distinct from those found throughout the rest of biology, including those of other hyperthermophilic archaea.« less

  5. Accumulation of Mannosylglycerate and Di-myo-Inositol-Phosphate by Pyrococcus furiosus in Response to Salinity and Temperature

    PubMed Central

    Martins, L. O.; Santos, H.

    1995-01-01

    (sup13)C and (sup1)H nuclear magnetic resonance spectroscopy was used to identify and quantify organic solutes accumulated by the hyperthermophilic archaeon Pyrococcus furiosus in response to temperature and salinity. Di-myo-inositol-phosphate and 2-O-(beta)-mannosylglycerate were the major organic solutes accumulated in these cells. The total intracellular organic solutes increased significantly in response either to an increase in temperature or to an increase in salinity, but (beta)-mannosylglycerate accumulated mainly at high salinities, whereas the concentration of di-myo-inositol-phosphate increased dramatically at supraoptimal growth temperatures. Glutamate was present at concentrations detectable by nuclear magnetic resonance only in cells grown in low-salinity media. The intracellular levels of K(sup+) are clearly dependent on the salinity of the medium, and the concentrations of this cation are high enough to counterbalance the negative charges of (beta)-mannosylglycerate and di-myo-inositol-phosphate in the cell. The results presented here together with those previously reported for Pyrococcus woesei (S. Scholz, J. Sonnenbichler, W. Schafer, and R. Hensel, FEBS Lett. 306:239-242, 1992) strongly support a role for di-myo-inositol-phosphate in thermoprotection. PMID:16535119

  6. Electronic, Magnetic, and Redox Properties of [MFe(3)S(4)] Clusters (M = Cd, Cu, Cr) in Pyrococcus furiosus Ferredoxin.

    PubMed

    Staples, Christopher R.; Dhawan, Ish K.; Finnegan, Michael G.; Dwinell, Derek A.; Zhou, Zhi Hao; Huang, Heshu; Verhagen, Marc F. J. M.; Adams, Michael W. W.; Johnson, Michael K.

    1997-12-01

    The ground- and excited-state properties of heterometallic [CuFe(3)S(4)](2+,+), [CdFe(3)S(4)](2+,+), and [CrFe(3)S(4)](2+,+) cubane clusters assembled in Pyrococcus furiosus ferredoxin have been investigated by the combination of EPR and variable-temperature/variable-field magnetic circular dichroism (MCD) studies. The results indicate Cd(2+) incorporation into [Fe(3)S(4)](0,-) cluster fragments to yield S = 2 [CdFe(3)S(4)](2+) and S = (5)/(2) [CdFe(3)S(4)](+) clusters and Cu(+) incorporation into [Fe(3)S(4)](+,0) cluster fragments to yield S = (1)/(2) [CuFe(3)S(4)](2+) and S = 2 [CuFe(3)S(4)](+) clusters. This is the first report of the preparation of cubane type [CrFe(3)S(4)](2+,+) clusters, and the combination of EPR and MCD results indicates S = 0 and S = (3)/(2) ground states for the oxidized and reduced forms, respectively. Midpoint potentials for the [CdFe(3)S(4)](2+,+), [CrFe(3)S(4)](2+,+), and [CuFe(3)S(4)](2+,+) couples, E(m) = -470 +/- 15, -440 +/- 10, and +190 +/- 10 mV (vs NHE), respectively, were determined by EPR-monitored redox titrations or direct electrochemistry at a glassy carbon electrode. The trends in redox potential, ground-state spin, and electron delocalization of [MFe(3)S(4)](2+,+) clusters in P. furiosus ferredoxin are discussed as a function of heterometal (M = Cr, Mn, Fe, Co, Ni, Cu, Zn, Cd, and Tl).

  7. Mannosylglycerate and Di-myo-Inositol Phosphate Have Interchangeable Roles during Adaptation of Pyrococcus furiosus to Heat Stress

    PubMed Central

    Esteves, Ana M.; Chandrayan, Sanjeev K.; McTernan, Patrick M.; Adams, Michael W. W.; Santos, Helena

    2014-01-01

    Marine hyperthermophiles accumulate small organic compounds, known as compatible solutes, in response to supraoptimal temperatures or salinities. Pyrococcus furiosus is a hyperthermophilic archaeon that grows optimally at temperatures near 100°C. This organism accumulates mannosylglycerate (MG) and di-myo-inositol phosphate (DIP) in response to osmotic and heat stress, respectively. It has been assumed that MG and DIP are involved in cell protection; however, firm evidence for the roles of these solutes in stress adaptation is still missing, largely due to the lack of genetic tools to produce suitable mutants of hyperthermophiles. Recently, such tools were developed for P. furiosus, making this organism an ideal target for that purpose. In this work, genes coding for the synthases in the biosynthetic pathways of MG and DIP were deleted by double-crossover homologous recombination. The growth profiles and solute patterns of the two mutants and the parent strain were investigated under optimal growth conditions and also at supraoptimal temperatures and NaCl concentrations. DIP was a suitable replacement for MG during heat stress, but substitution of MG for DIP and aspartate led to less efficient growth under conditions of osmotic stress. The results suggest that the cascade of molecular events leading to MG synthesis is tuned for osmotic adjustment, while the machinery for induction of DIP synthesis responds to either stress agent. MG protects cells against heat as effectively as DIP, despite the finding that the amount of DIP consistently increases in response to heat stress in the nine (hyper)thermophiles examined thus far. PMID:24795373

  8. DNA targeting by the type I-G and type I-A CRISPR–Cas systems of Pyrococcus furiosus

    PubMed Central

    Elmore, Joshua; Deighan, Trace; Westpheling, Jan; Terns, Rebecca M.; Terns, Michael P.

    2015-01-01

    CRISPR–Cas systems silence plasmids and viruses in prokaryotes. CRISPR–Cas effector complexes contain CRISPR RNAs (crRNAs) that include sequences captured from invaders and direct CRISPR-associated (Cas) proteins to destroy corresponding invader nucleic acids. Pyrococcus furiosus (Pfu) harbors three CRISPR–Cas immune systems: a Cst (Type I-G) system with an associated Cmr (Type III-B) module at one locus, and a partial Csa (Type I-A) module (lacking known invader sequence acquisition and crRNA processing genes) at another locus. The Pfu Cmr complex cleaves complementary target RNAs, and Csa systems have been shown to target DNA, while the mechanism by which Cst complexes silence invaders is unknown. In this study, we investigated the function of the Cst as well as Csa system in Pfu strains harboring a single CRISPR–Cas system. Plasmid transformation assays revealed that the Cst and Csa systems both function by DNA silencing and utilize similar flanking sequence information (PAMs) to identify invader DNA. Silencing by each system specifically requires its associated Cas3 nuclease. crRNAs from the 7 shared CRISPR loci in Pfu are processed for use by all 3 effector complexes, and Northern analysis revealed that individual effector complexes dictate the profile of mature crRNA species that is generated. PMID:26519471

  9. Structures of the Signal Recognition Particle Receptor From the Archaeon Pyrococcus Furiosus: Implications for the Targeting Step at the Membrane

    SciTech Connect

    Egea, P.F.; Tsuruta, H.; Leon, G.P.de; Napetschnig, J.; Walter, P.; Stroud, R.M.

    2009-05-18

    In all organisms, a ribonucleoprotein called the signal recognition particle (SRP) and its receptor (SR) target nascent proteins from the ribosome to the translocon for secretion or membrane insertion. We present the first X-ray structures of an archeal FtsY, the receptor from the hyper-thermophile Pyrococcus furiosus (Pfu), in its free and GDP {center_dot} magnesium-bound forms. The highly charged N-terminal domain of Pfu-FtsY is distinguished by a long N-terminal helix. The basic charges on the surface of this helix are likely to regulate interactions at the membrane. A peripheral GDP bound near a regulatory motif could indicate a site of interaction between the receptor and ribosomal or SRP RNAs. Small angle X-ray scattering and analytical ultracentrifugation indicate that the crystal structure of Pfu-FtsY correlates well with the average conformation in solution. Based on previous structures of two sub-complexes, we propose a model of the core of archeal and eukaryotic SRP {center_dot} SR targeting complexes.

  10. Expression and Characterization of a Novel Glycerophosphodiester Phosphodiesterase from Pyrococcus furiosus DSM 3638 That Possesses Lysophospholipase D Activity.

    PubMed

    Wang, Fanghua; Lai, Linhui; Liu, Yanhua; Yang, Bo; Wang, Yonghua

    2016-01-01

    Glycerophosphodiester phosphodiesterases (GDPD) are enzymes which degrade various glycerophosphodiesters to produce glycerol-3-phosphate and the corresponding alcohol moiety. Apart from this, a very interesting finding is that this enzyme could be used in the degradation of toxic organophosphorus esters, which has resulted in much attention on the biochemical and application research of GDPDs. In the present study, a novel GDPD from Pyrococcus furiosus DSM 3638 (pfGDPD) was successfully expressed in Escherichia coli and biochemically characterized. This enzyme hydrolyzed bis(p-nitrophenyl) phosphate, one substrate analogue of organophosphorus diester, with an optimal reaction temperature 55 °C and pH 8.5. The activity of pfGDPD was strongly dependent on existing of bivalent cations. It was strongly stimulated by Mn(2+) ions, next was Co(2+) and Ni(2+) ions. Further investigations were conducted on its substrate selectivity towards different phospholipids. The results indicated that except of glycerophosphorylcholine (GPC), this enzyme also possessed lysophospholipase D activity toward both sn1-lysophosphatidylcholine (1-LPC) and sn2-lysophosphatidylcholine (2-LPC). Higher activity was found for 1-LPC than 2-LPC; however, no hydrolytic activity was found for phosphatidylcholine (PC). Molecular docking based on the 3D-modeled structure of pfGDPD was conducted in order to provide a structural foundation for the substrate selectivity. PMID:27248999

  11. Structures of the Signal Recognition Particle Receptor from the Archaeon Pyrococcus furiosus: Implications for the Targeting Step at the Membrane

    PubMed Central

    Egea, Pascal F.; Tsuruta, Hiro; de Leon, Gladys P.; Napetschnig, Johanna; Walter, Peter; Stroud, Robert M.

    2008-01-01

    In all organisms, a ribonucleoprotein called the signal recognition particle (SRP) and its receptor (SR) target nascent proteins from the ribosome to the translocon for secretion or membrane insertion. We present the first X-ray structures of an archeal FtsY, the receptor from the hyper-thermophile Pyrococcus furiosus (Pfu), in its free and GDP•magnesium-bound forms. The highly charged N-terminal domain of Pfu-FtsY is distinguished by a long N-terminal helix. The basic charges on the surface of this helix are likely to regulate interactions at the membrane. A peripheral GDP bound near a regulatory motif could indicate a site of interaction between the receptor and ribosomal or SRP RNAs. Small angle X-ray scattering and analytical ultracentrifugation indicate that the crystal structure of Pfu-FtsY correlates well with the average conformation in solution. Based on previous structures of two sub-complexes, we propose a model of the core of archeal and eukaryotic SRP•SR targeting complexes. PMID:18978942

  12. Improving the Thermostability and Optimal Temperature of a Lipase from the Hyperthermophilic Archaeon Pyrococcus furiosus by Covalent Immobilization

    PubMed Central

    Branco, Roberta V.; Gutarra, Melissa L. E.; Freire, Denise M. G.; Almeida, Rodrigo V.; Palomo, Jose M.

    2015-01-01

    A recombinant thermostable lipase (Pf2001Δ60) from the hyperthermophilic Archaeon Pyrococcus furiosus (PFUL) was immobilized by hydrophobic interaction on octyl-agarose (octyl PFUL) and by covalent bond on aldehyde activated-agarose in the presence of DTT at pH = 7.0 (one-point covalent attachment) (glyoxyl-DTT PFUL) and on glyoxyl-agarose at pH 10.2 (multipoint covalent attachment) (glyoxyl PFUL). The enzyme's properties, such as optimal temperature and pH, thermostability, and selectivity, were improved by covalent immobilization. The highest enzyme stability at 70°C for 48 h incubation was achieved for glyoxyl PFUL (around 82% of residual activity), whereas glyoxyl-DTT PFUL maintained around 69% activity, followed by octyl PFUL (27% remaining activity). Immobilization on glyoxyl-agarose improved the optimal temperature to 90°C, while the optimal temperature of octyl PFUL was 70°C. Also, very significant changes in activity with different substrates were found. In general, the covalent bond derivatives were more active than octyl PFUL. The E value also depended substantially on the derivative and the conditions used. It was observed that the reaction of glyoxyl-DTT PFUL using methyl mandelate as a substrate at pH 7 presented the best results for enantioselectivity (E = 22) and enantiomeric excess (ee (%) = 91). PMID:25839031

  13. Expression and Characterization of a Novel Glycerophosphodiester Phosphodiesterase from Pyrococcus furiosus DSM 3638 That Possesses Lysophospholipase D Activity

    PubMed Central

    Wang, Fanghua; Lai, Linhui; Liu, Yanhua; Yang, Bo; Wang, Yonghua

    2016-01-01

    Glycerophosphodiester phosphodiesterases (GDPD) are enzymes which degrade various glycerophosphodiesters to produce glycerol-3-phosphate and the corresponding alcohol moiety. Apart from this, a very interesting finding is that this enzyme could be used in the degradation of toxic organophosphorus esters, which has resulted in much attention on the biochemical and application research of GDPDs. In the present study, a novel GDPD from Pyrococcus furiosus DSM 3638 (pfGDPD) was successfully expressed in Escherichia coli and biochemically characterized. This enzyme hydrolyzed bis(p-nitrophenyl) phosphate, one substrate analogue of organophosphorus diester, with an optimal reaction temperature 55 °C and pH 8.5. The activity of pfGDPD was strongly dependent on existing of bivalent cations. It was strongly stimulated by Mn2+ ions, next was Co2+ and Ni2+ ions. Further investigations were conducted on its substrate selectivity towards different phospholipids. The results indicated that except of glycerophosphorylcholine (GPC), this enzyme also possessed lysophospholipase D activity toward both sn1-lysophosphatidylcholine (1-LPC) and sn2-lysophosphatidylcholine (2-LPC). Higher activity was found for 1-LPC than 2-LPC; however, no hydrolytic activity was found for phosphatidylcholine (PC). Molecular docking based on the 3D-modeled structure of pfGDPD was conducted in order to provide a structural foundation for the substrate selectivity. PMID:27248999

  14. Physiological aggregation of maltodextrin phosphorylase from Pyrococcus furiosus and its application in a process of batch starch degradation to alpha-D-glucose-1-phosphate.

    PubMed

    Nahálka, Jozef

    2008-04-01

    Maltodextrin phosphorylase from Pyrococcus furiosus (PF1535) was fused with the cellulose-binding domain of Clostridium cellulovorans serving as an aggregation module. After molecular cloning of the corresponding gene fusion construct and controlled expression in Escherichia coli BL21, 83% of total maltodextrin phosphorylase activity (0.24 U/mg of dry cell weight) was displayed in active inclusion bodies. These active inclusion bodies were easily isolated by nonionic detergent treatment and directly used for maltodextrin conversion to alpha-D-glucose-1-phosphate in a repetitive batch mode. Only 10% of enzyme activity was lost after ten conversion cycles at optimum conditions.

  15. MAGGIE Component 1: Identification and Purification of Native and Recombinant Multiprotein Complexes and Modified Proteins from Pyrococcus furiosus

    SciTech Connect

    Adams, Michael W.; W. W. Adams, Michael

    2014-01-07

    Virtualy all cellular processes are carried out by dynamic molecular assemblies or multiprotein complexes (PCs), the composition of which is largely unknown. Structural genomics efforts have demonstrated that less than 25% of the genes in a given prokaryotic genome will yield stable, soluble proteins when expressed using a one-ORF-at-a-time approach. We proposed that much of the remaining 75% of the genes encode proteins that are part of multiprotein complexes or are modified post-translationally, for example, with metals. The problem is that PCs and metalloproteins (MPs) cannot be accurately predicted on a genome-wide scale. The only solution to this dilemma is to experimentally determine PCs and MPs in biomass of a model organism and to develop analytical tools that can then be applied to the biomass of any other organism. In other words, organisms themselves must be analyzed to identify their PCs and MPs: “native proteomes” must be determined. This information can then be utilized to design multiple ORF expression systems to produce recombinant forms of PCs and MPs. Moreover, the information and utility of this approach can be enhanced by using a hyperthermophile, one that grows optimally at 100°C, as a model organism. By analyzing the native proteome at close to 100 °C below the optimum growth temperature, we will trap reversible and dynamic complexes, thereby enabling their identification, purification, and subsequent characterization. The model organism for the current study is Pyrococcus furiosus, a hyperthermophilic archaeon that grows optimally at 100°C. It is grown up to 600-liter scale and kg quantities of biomass are available. In this project we identified native PCs and MPs using P. furiosus biomass (with MS/MS analyses to identify proteins by component 4). In addition, we provided samples of abundant native PCs and MPs for structural characterization (using SAXS by component 5). We also designed and evaluated generic bioinformatics and

  16. The L7Ae protein binds to two kink-turns in the Pyrococcus furiosus RNase P RNA.

    PubMed

    Lai, Stella M; Lai, Lien B; Foster, Mark P; Gopalan, Venkat

    2014-12-01

    The RNA-binding protein L7Ae, known for its role in translation (as part of ribosomes) and RNA modification (as part of sn/oRNPs), has also been identified as a subunit of archaeal RNase P, a ribonucleoprotein complex that employs an RNA catalyst for the Mg(2+)-dependent 5' maturation of tRNAs. To better understand the assembly and catalysis of archaeal RNase P, we used a site-specific hydroxyl radical-mediated footprinting strategy to pinpoint the binding sites of Pyrococcus furiosus (Pfu) L7Ae on its cognate RNase P RNA (RPR). L7Ae derivatives with single-Cys substitutions at residues in the predicted RNA-binding interface (K42C/C71V, R46C/C71V, V95C/C71V) were modified with an iron complex of EDTA-2-aminoethyl 2-pyridyl disulfide. Upon addition of hydrogen peroxide and ascorbate, these L7Ae-tethered nucleases were expected to cleave the RPR at nucleotides proximal to the EDTA-Fe-modified residues. Indeed, footprinting experiments with an enzyme assembled with the Pfu RPR and five protein cofactors (POP5, RPP21, RPP29, RPP30 and L7Ae-EDTA-Fe) revealed specific RNA cleavages, localizing the binding sites of L7Ae to the RPR's catalytic and specificity domains. These results support the presence of two kink-turns, the structural motifs recognized by L7Ae, in distinct functional domains of the RPR and suggest testable mechanisms by which L7Ae contributes to RNase P catalysis.

  17. Structural Basis of Thermal Stability of the Tungsten Cofactor Synthesis Protein MoaB from Pyrococcus furiosus

    PubMed Central

    Havarushka, Nastassia; Fischer-Schrader, Katrin; Lamkemeyer, Tobias; Schwarz, Guenter

    2014-01-01

    Molybdenum and tungsten cofactors share a similar pterin-based scaffold, which hosts an ene-dithiolate function being essential for the coordination of either molybdenum or tungsten. The biosynthesis of both cofactors involves a multistep pathway, which ends with the activation of the metal binding pterin (MPT) by adenylylation before the respective metal is incorporated. In the hyperthermophilic organism Pyrococcus furiosus, the hexameric protein MoaB (PfuMoaB) has been shown to catalyse MPT-adenylylation. Here we determined the crystal structure of PfuMoaB at 2.5 Å resolution and identified key residues of α3-helix mediating hexamer formation. Given that PfuMoaB homologues from mesophilic organisms form trimers, we investigated the impact on PfuMoaB hexamerization on thermal stability and activity. Using structure-guided mutagenesis, we successfully disrupted the hexamer interface in PfuMoaB. The resulting PfuMoaB-H3 variant formed monomers, dimers and trimers as determined by size exclusion chromatography. Circular dichroism spectroscopy as well as chemical cross-linking coupled to mass spectrometry confirmed a wild-type-like fold of the protomers as well as inter-subunits contacts. The melting temperature of PfuMoaB-H3 was found to be reduced by more than 15°C as determined by differential scanning calorimetry, thus demonstrating hexamerization as key determinant for PfuMoaB thermal stability. Remarkably, while a loss of activity at temperatures higher than 50°C was observed in the PfuMoaB-H3 variant, at lower temperatures, we determined a significantly increased catalytic activity. The latter suggests a gain in conformational flexibility caused by the disruption of the hexamerization interface. PMID:24465852

  18. Preparation of lactose-free pasteurized milk with a recombinant thermostable β-glucosidase from Pyrococcus furiosus

    PubMed Central

    2013-01-01

    Background Lactose intolerance is a common health concern causing gastrointestinal symptoms and avoidance of dairy products by afflicted individuals. Since milk is a primary source of calcium and vitamin D, lactose intolerant individuals often obtain insufficient amounts of these nutrients which may lead to adverse health outcomes. Production of lactose-free milk can provide a solution to this problem, although it requires use of lactase from microbial sources and increases potential for contamination. Use of thermostable lactase enzymes can overcome this issue by functioning under pasteurization conditions. Results A thermostable β-glucosidase gene from Pyrococcus furiosus was cloned in frame with the Saccharomyces cerecisiae a-factor secretory signal and expressed in Pichia pastoris strain X-33. The recombinant enzyme was purified by a one-step method of weak anion exchange chromatography. The optimum temperature and pH for this β-glucosidase activity was 100°C and pH 6.0, respectively. The enzyme activity was not significantly inhibited by Ca2+. We tested the additive amount, hydrolysis time, and the influence of glucose on the enzyme during pasteurization and found that the enzyme possessed a high level of lactose hydrolysis in milk that was not obviously influenced by glucose. Conclusions The thermostablity of this recombinant β-glucosidase, combined with its neutral pH activity and favorable temperature activity optima, suggest that this enzyme is an ideal candidate for the hydrolysis of lactose in milk, and it would be suitable for application in low-lactose milk production during pasteurization. PMID:24053641

  19. Structural basis of thermal stability of the tungsten cofactor synthesis protein MoaB from Pyrococcus furiosus.

    PubMed

    Havarushka, Nastassia; Fischer-Schrader, Katrin; Lamkemeyer, Tobias; Schwarz, Guenter

    2014-01-01

    Molybdenum and tungsten cofactors share a similar pterin-based scaffold, which hosts an ene-dithiolate function being essential for the coordination of either molybdenum or tungsten. The biosynthesis of both cofactors involves a multistep pathway, which ends with the activation of the metal binding pterin (MPT) by adenylylation before the respective metal is incorporated. In the hyperthermophilic organism Pyrococcus furiosus, the hexameric protein MoaB (PfuMoaB) has been shown to catalyse MPT-adenylylation. Here we determined the crystal structure of PfuMoaB at 2.5 Å resolution and identified key residues of α3-helix mediating hexamer formation. Given that PfuMoaB homologues from mesophilic organisms form trimers, we investigated the impact on PfuMoaB hexamerization on thermal stability and activity. Using structure-guided mutagenesis, we successfully disrupted the hexamer interface in PfuMoaB. The resulting PfuMoaB-H3 variant formed monomers, dimers and trimers as determined by size exclusion chromatography. Circular dichroism spectroscopy as well as chemical cross-linking coupled to mass spectrometry confirmed a wild-type-like fold of the protomers as well as inter-subunits contacts. The melting temperature of PfuMoaB-H3 was found to be reduced by more than 15 °C as determined by differential scanning calorimetry, thus demonstrating hexamerization as key determinant for PfuMoaB thermal stability. Remarkably, while a loss of activity at temperatures higher than 50 °C was observed in the PfuMoaB-H3 variant, at lower temperatures, we determined a significantly increased catalytic activity. The latter suggests a gain in conformational flexibility caused by the disruption of the hexamerization interface. PMID:24465852

  20. Influence of ionic liquid cosolvent on transgalactosylation reactions catalyzed by thermostable beta-glycosylhydrolase CelB from Pyrococcus Furiosus.

    PubMed

    Lang, Markus; Kamrat, Thomas; Nidetzky, Bernd

    2006-12-20

    The synthesis of glycosides by enzymatic transglycosylation is a kinetically controlled reaction performed in the context of a non-favorable thermodynamic equilibrium. An unreactive organic cosolvent which increases the selectivity of the enzyme for glycosyl transfer to the acceptor nucleophile compared with water (Ksel) could improve maximum product yield. Here we report on the effect of the ionic liquid 1,3-dimethylimidazoliummethylsulfate on hydrolase and transferase activities of the hyperthermostable beta-glycosidase CelB from the archaeon Pyrococcus furiosus. CelB retained full catalytic efficiency for lactose hydrolysis at 80 degrees C in a 50% (by vol.) solution of ionic liquid in sodium citrate buffer, pH 5.5. It was inactive but not irreversibly denatured at 70% ionic liquid. Using lactose (0.15 M) as galactosyl donor, values of Ksel for a representative series of eight acceptor alcohols were determined in kinetic assays at 80 degrees C and found to increase between 1.3-fold (D-xylose) and 3.1-fold (glycerol) in 45% ionic liquid. Enhancement of Ksel was dependent on ionic liquid concentration and higher than expected from the decrease in water activity caused by the cosolvent. Experimental molar ratios of D-glucose and D-galactose produced during enzymatic conversion of lactose (75-150 mM) in the presence of D-xylose (0.5 M) or glycerol (0.5 M) showed excellent agreement with predictions based on Ksel values and confirm a significant, yet moderate effect of 45% ionic liquid on increasing the yield of D-galactoside product, by < or = 10%.

  1. The role of TrmB and TrmB-like transcriptional regulators for sugar transport and metabolism in the hyperthermophilic archaeon Pyrococcus furiosus.

    PubMed

    Lee, Sung-Jae; Surma, Melanie; Hausner, Winfried; Thomm, Michael; Boos, Winfried

    2008-09-01

    TrmB of Pyrococcus furiosus was discovered as the trehalose/maltose-specific repressor for the genes encoding the trehalose/maltose high-affinity ABC transporter (the TM system). TrmB also represses the genes encoding the high affinity maltodextrin-specific ABC transporter (the MD system) with maltodextrin and sucrose as inducers. In addition, TrmB binds glucose leading to an increased repression of both, the TM and the MD system. Thus, TrmB recognizes different promoters and depending on the promoter it will be activated or inactivated for promoter binding by different sugar effectors. The TrmB-like protein TrmBL1 of P. furiosus is a global regulator and recognizes preferentially, but not exclusively, the TGM (for Thermococcales-glycolytic motif) sequence that is found upstream of the MD system as well as of genes encoding enzymes involved in the glycolytic and the gluconeogenic pathway. It responds to maltose and maltotriose as inducers and functions as repressor for the genes encoding the MD system and glycolytic enzymes, but as activator for genes encoding gluconeogenic enzymes. The TrmB-like protein TrmBL2 of P. furiosus lacks the sugar-binding domain that has been determined in TrmB. It recognizes the MD promoter, but not all TGM harboring promoters. It is evolutionary the most conserved among the Thermococcales. The regulatory range of TrmBL2 remains unclear.

  2. The ferredoxin-dependent conversion of glyceraldehyde-3-phosphate in the hyperthermophilic archaeon Pyrococcus furiosus represents a novel site of glycolytic regulation.

    PubMed

    van der Oost, J; Schut, G; Kengen, S W; Hagen, W R; Thomm, M; de Vos, W M

    1998-10-23

    The fermentative conversion of glucose in anaerobic hyperthermophilic Archaea is a variant of the classical Embden-Meyerhof pathway found in Bacteria and Eukarya. A major difference of the archaeal glycolytic pathway concerns the conversion of glyceraldehyde-3-phosphate. In the hyperthermophilic archaeon Pyrococcus furiosus, this reaction is catalyzed by an unique enzyme, glyceraldehyde-3-phosphate ferredoxin oxidoreductase (GAPOR). Here, we report the isolation, characterization, and transcriptional analysis of the GAPOR-encoding gene. GAPOR is related to a family of ferredoxin-dependent tungsten enzymes in (hyper)thermophilic Archaea and, in addition, to a hypothetical protein in Escherichia coli. Electron paramagnetic resonance analysis of the purified P. furiosus GAPOR protein confirms the anticipated involvement of tungsten in catalysis. During glycolysis in P. furiosus, GAPOR gene expression is induced, whereas the activity of glyceraldehyde-3-phosphate dehydrogenase is repressed. It is discussed that this unprecedented unidirectional reaction couple in the pyrococcal glycolysis and gluconeogenesis gives rise to a novel site of glycolytic regulation that might be widespread among Archaea.

  3. Enhancing Heat Tolerance of the Little Dogwood Cornus canadensis L. f. with Introduction of a Superoxide Reductase Gene from the Hyperthermophilic Archaeon Pyrococcus furiosus

    PubMed Central

    Geng, Xing-Min; Liu, Xiang; Ji, Mikyoung; Hoffmann, William A.; Grunden, Amy; Xiang, Qiu-Yun J.

    2016-01-01

    Production of reactive oxygen species (ROS) can be accelerated under various biotic and abiotic stresses causing lipid peroxidation, protein degradation, enzyme inactivation, and DNA damage. Superoxide reductase (SOR) is a novel antioxidant enzyme from Pyrococcus furiosus and is employed by this anaerobic hyperthermophilic archaeon for efficient detoxification of ROS. In this study, SOR was introduced into a flowering plant Cornus canadensis to enhance its heat tolerance and reduce heat induced damage. A fusion construct of the SOR gene and Green Fluorescent Protein gene (GFP) was introduced into C. canadensis using Agrobacterium-mediated transformation. Heat tolerance of the GFP-SOR expressing transgenic plants was investigated by observing morphological symptoms of heat injury and by examining changes in photosynthesis, malondialdehyde (MDA), and proline levels in the plants. Our results indicate that the expression of the P. furiosus SOR gene in the transgenic plants alleviated lipid peroxidation of cell membranes and photoinhibition of PS II, and decreased the accumulation of proline at 40°C. After a series of exposures to increasing temperatures, the SOR transgenic plants remained healthy and green whereas most of the non-transgenic plants dried up and were unable to recover. While it had previously been reported that expression of SOR in Arabidopsis enhanced heat tolerance, this is the first report of the successful demonstration of improved heat tolerance in a non-model plant resulting from the introduction of P. furiosus SOR. The study demonstrates the potential of SOR for crop improvement and that inherent limitations of plant heat tolerance can be ameliorated with P. furiosus SOR. PMID:26858741

  4. Enhancing Heat Tolerance of the Little Dogwood Cornus canadensis L. f. with Introduction of a Superoxide Reductase Gene from the Hyperthermophilic Archaeon Pyrococcus furiosus.

    PubMed

    Geng, Xing-Min; Liu, Xiang; Ji, Mikyoung; Hoffmann, William A; Grunden, Amy; Xiang, Qiu-Yun J

    2016-01-01

    Production of reactive oxygen species (ROS) can be accelerated under various biotic and abiotic stresses causing lipid peroxidation, protein degradation, enzyme inactivation, and DNA damage. Superoxide reductase (SOR) is a novel antioxidant enzyme from Pyrococcus furiosus and is employed by this anaerobic hyperthermophilic archaeon for efficient detoxification of ROS. In this study, SOR was introduced into a flowering plant Cornus canadensis to enhance its heat tolerance and reduce heat induced damage. A fusion construct of the SOR gene and Green Fluorescent Protein gene (GFP) was introduced into C. canadensis using Agrobacterium-mediated transformation. Heat tolerance of the GFP-SOR expressing transgenic plants was investigated by observing morphological symptoms of heat injury and by examining changes in photosynthesis, malondialdehyde (MDA), and proline levels in the plants. Our results indicate that the expression of the P. furiosus SOR gene in the transgenic plants alleviated lipid peroxidation of cell membranes and photoinhibition of PS II, and decreased the accumulation of proline at 40°C. After a series of exposures to increasing temperatures, the SOR transgenic plants remained healthy and green whereas most of the non-transgenic plants dried up and were unable to recover. While it had previously been reported that expression of SOR in Arabidopsis enhanced heat tolerance, this is the first report of the successful demonstration of improved heat tolerance in a non-model plant resulting from the introduction of P. furiosus SOR. The study demonstrates the potential of SOR for crop improvement and that inherent limitations of plant heat tolerance can be ameliorated with P. furiosus SOR.

  5. Evidence for the operation of a novel Embden-Meyerhof pathway that involves ADP-dependent kinases during sugar fermentation by Pyrococcus furiosus.

    PubMed

    Kengen, S W; de Bok, F A; van Loo, N D; Dijkema, C; Stams, A J; de Vos, W M

    1994-07-01

    The main pathway for the fermentation of maltose or cellobiose by the hyperthermophile Pyrococcus furiosus was investigated by in vivo NMR and by enzyme measurements. Addition of [1-13C]glucose to cell suspensions resulted in the formation of C2-labeled acetate and C3-labeled alanine. No label was recovered in CO2 or HCO3-. In the presence of [3-13C]glucose, the label ended up in the C1 atom of alanine and in HCO3- and CO2. These labeling patterns indicate that glucose is converted along an Embden-Meyerhof pathway, and they disagree with the previously proposed nonphosphorylated Entner-Doudoroff pathway (pyroglycolysis). The NMR data were supported by enzyme measurements. Hexokinase (8.7 units/mg), phosphoglucose isomerase (6.8 units/mg), phosphofructokinase (0.81 unit/mg), and aldolase (0.26 unit/mg) were present in cell-free extracts (specific activities at 90 degrees C). Remarkably, the two kinases required ADP as the phosphoryl group donor instead of ATP. No activity was found with pyrophosphate. These are the first descriptions of ADP-dependent (AMP-forming) kinases to date. Since P. furiosus is a phylogenetically ancient organism, these enzymes may represent an ancestral kind of metabolism.

  6. Exploitation of the S-layer self-assembly system for site directed immobilization of enzymes demonstrated for an extremophilic laminarinase from Pyrococcus furiosus

    PubMed Central

    Tschiggerl, Helga; Breitwieser, Andreas; de Roo, Guy; Verwoerd, Theo; Scḧaffer, Christina; Sleytr, Uwe B.

    2015-01-01

    A fusion protein based on the S-layer protein SbpA from Bacillus sphaericus CCM 2177 and the enzyme laminarinase (LamA) from Pyrococcus furiosus was designed and overexpressed in Escherichia coli. Due to the construction principle, the S-layer fusion protein fully retained the self-assembly capability of the S-layer moiety, while the catalytic domain of LamA remained exposed at the outer surface of the formed protein lattice. The enzyme activity of the S-layer fusion protein monolayer obtained upon recrystallization on silicon wafers, glass slides and different types of polymer membranes was determined colorimetrically and related to the activity of sole LamA that has been immobilized with conventional techniques. LamA aligned within the S-layer fusion protein lattice in a periodic and orientated fashion catalyzed twice the glucose release from the laminarin polysaccharide substrate in comparison to the randomly immobilized enzyme. In combination with the good shelf-life and the high resistance towards temperature and diverse chemicals, these novel composites are regarded a promising approach for site-directed enzyme immobilization. PMID:18035441

  7. 'Super-perfect' enzymes: Structural stabilities and activities of recombinant triose phosphate isomerases from Pyrococcus furiosus and Thermococcus onnurineus produced in Escherichia coli.

    PubMed

    Sharma, Prerna; Guptasarma, Purnananda

    2015-05-01

    Triose phosphate isomerases (TIMs) are considered to be 'kinetically perfect' enzymes, limited in their activity only by the rates of diffusion of substrate and product molecules. Most studies conducted thus far have been on mesophile-derived TIMs. Here, we report studies of two extremophile-derived TIMs produced in Escherichia coli: (i) TonTIM, sourced from the genome of the thermophile archaeon, Thermococcus onnurineus, and (ii) PfuTIM, sourced from the genome of the hyperthermophile archaeon, Pyrococcus furiosus (PfuTIM). Although these enzymes are presumed to have evolved to function optimally at temperatures close to the boiling point of water, we find that TonTIM and PfuTIM display second-order rate-constants of activity (k(cat)/K(m) values) comparable to mesophile-derived TIMs, at 25 °C. At 90 °C, TonTIM and PfuTIM reach maximum velocities of reaction of ∼ 10(6)-10(7) μmol/s/mg, and display k(cat)/K(m) values in the range of ∼ 10(10)-10(11) M(-1) s(-1), which are three orders of magnitude higher than those reported for mesophile TIMs. Further, the two enzymes display no signs of having undergone any structural unfolding at 90 °C. Such enzymes could thus probably be called 'super-perfect' enzymes. PMID:25824038

  8. Uncovering the stoichiometry of Pyrococcus furiosus RNase P, a multi-subunit catalytic ribonucleoprotein complex, by surface-induced dissociation and ion mobility mass spectrometry.

    PubMed

    Ma, Xin; Lai, Lien B; Lai, Stella M; Tanimoto, Akiko; Foster, Mark P; Wysocki, Vicki H; Gopalan, Venkat

    2014-10-20

    We demonstrate that surface-induced dissociation (SID) coupled with ion mobility mass spectrometry (IM-MS) is a powerful tool for determining the stoichiometry of a multi-subunit ribonucleoprotein (RNP) complex assembled in a solution containing Mg(2+). We investigated Pyrococcus furiosus (Pfu) RNase P, an archaeal RNP that catalyzes tRNA 5' maturation. Previous step-wise, Mg(2+)-dependent reconstitutions of Pfu RNase P with its catalytic RNA subunit and two interacting protein cofactor pairs (RPP21⋅RPP29 and POP5⋅RPP30) revealed functional RNP intermediates en route to the RNase P enzyme, but provided no information on subunit stoichiometry. Our native MS studies with the proteins showed RPP21⋅RPP29 and (POP5⋅RPP30)2 complexes, but indicated a 1:1 composition for all subunits when either one or both protein complexes bind the cognate RNA. These results highlight the utility of SID and IM-MS in resolving conformational heterogeneity and yielding insights on RNP assembly.

  9. Crystallization and preliminary X-ray characterization of a ferritin from the hyperthermophilic archaeon and anaerobe Pyrococcus furiosus

    SciTech Connect

    Matias, Pedro M.; Tatur, Jana; Carrondo, Maria Arménia; Hagen, Wilfred R.

    2005-05-01

    Ferritin from P. furiosus crystallizes in space group C222{sub 1}, with unit-cell parameters a = 258.1, b = 340.1, c = 266.5 Å and 36 monomers in the asymmetric unit, corresponding to one and a half 24-mers. Crystals of the title protein have been produced and preliminary structural analysis has been carried out. The crystals belong to the orthorhombic space group C222{sub 1}, with unit-cell parameters a = 258.1, b = 340.1, c = 266.5 Å. The protein forms a 24-mer of 20 kDa subunits, which assemble with 432 non-crystallographic symmetry. A total of 36 monomers are found in the asymmetric unit, corresponding to one and a half 24-mers.

  10. Characterization of the TrmB-like protein, PF0124, a TGM-recognizing global transcriptional regulator of the hyperthermophilic archaeon Pyrococcus furiosus.

    PubMed

    Lee, Sung-Jae; Surma, Melanie; Seitz, Sabine; Hausner, Winfried; Thomm, Michael; Boos, Winfried

    2007-07-01

    The characterization of the transcriptional regulator TrmBL1 of the hyperthermophilic archaeon Pyrococcus furiosus, homologous to TrmB (transcriptional regulator of the maltose system), was studied. The genome of P. furiosus contains three TrmB paralogues. One of the TrmB-like proteins (TrmBL), PF0124 (TrmBL1), was analysed in more detail. It regulated the expression of the genes encoding enzymes of the glycolytic pathway as well as the maltodextrin (MD) ABC transporter. By molecular sieve chromatography, purified TrmBL1 behaved at ambient temperature as a tetramer of 148.8 kDa. In the presence of 1 mM maltotriose or 5 mM maltose TrmBL1 formed octamers. As shown by electrophoretic mobility shift assay (EMSA) TrmBL1 was found to bind the MD (maltodextrin ABC transport genes) promoter DNA with sixfold higher binding affinity (K(d) 0.2 microM) than to the trehalose/maltose ABC transporter (TM) promoter (K(d) 1.2 microM). Maltotriose and maltose interfered in these assays indicating inducer function. In vitro transcription assays using purified transcription components corroborated the data obtained with EMSA and showed inhibition of transcription of the MD promoter by TrmBL1. Recently, van de Werken et al. (FEMS Microbiol Lett 2006; 260: 69-76) identified TGM, a conserved sequence (Thermococcales-Glycolytic-Motif) upstream of genes encoding glycolytic enzymes and the MD ABC transporter. The position of TGM is invariably located downstream of the BRE-TATA box and overlapping the transcription start site on each promoter. By footprint analysis TrmBL1 was found to recognize the TGM sequence in several TGM-containing promoter sequences. We identified the recognition helix in TrmBL1 revealing tyrosine (Y49) to be essential for target DNA binding. However, the TGM motif was not essential for TrmBL1 binding. We conclude that TrmBL1 is a global sugar-sensing transcriptional regulator controlling the genes of transport systems and of sugar-metabolizing enzymes.

  11. Pyrococcus furiosus flagella: biochemical and transcriptional analyses identify the newly detected flaB0 gene to encode the major flagellin

    PubMed Central

    Näther-Schindler, Daniela J.; Schopf, Simone; Bellack, Annett; Rachel, Reinhard; Wirth, Reinhard

    2014-01-01

    We have described previously that the flagella of the Euryarchaeon Pyrococcus furiosus are multifunctional cell appendages used for swimming, adhesion to surfaces and formation of cell-cell connections. Here, we characterize these organelles with respect to their biochemistry and transcription. Flagella were purified by shearing from cells followed by CsCl-gradient centrifugation and were found to consist mainly of a ca. 30 kDa glycoprotein. Polymerization studies of denatured flagella resulted in an ATP-independent formation of flagella-like filaments. The N-terminal sequence of the main flagellin was determined by Edman degradation, but none of the genes in the complete genome code for a protein with that N-terminus. Therefore, we resequenced the respective region of the genome, thereby discovering that the published genome sequence is not correct. A total of 771 bp are missing in the data base, resulting in the correction of the previously unusual N-terminal sequence of flagellin FlaB1 and in the identification of a third flagellin. To keep in line with the earlier nomenclature we call this flaB0. Very interestingly, the previously not identified flaB0 codes for the major flagellin. Transcriptional analyses of the revised flagellar operon identified various different cotranscripts encoding only a single protein in case of FlaB0 and FlaJ or up to five proteins (FlaB0-FlaD). Analysing the RNA of cells from different growth phases, we found that the length and number of detected cotranscript increased over time suggesting that the flagellar operon is transcribed mostly in late exponential and stationary growth phase. PMID:25566211

  12. Hydrogenase of the hyperthermophile Pyrococcus furiosus is an elemental sulfur reductase or sulfhydrogenase: Evidence for a sulfur-reducing hydrogenase ancestor

    SciTech Connect

    Ma, K.; Adams, M.W.W. ); Schicho, R.N. ); Kelly, R.M. )

    1993-06-01

    Microorganisms growing near and above 100[degrees]C have recently been discovered near shallow and deep sea hydrothermal vents. Most are obligately dependent upon the reduction of elemental sulfur (S[sup 0]) to hydrogen sulfide (H[sub 2]S) for optimal growth, even though S[sup 0] reduction readily occurs abiotically at their growth temperatures. The sulfur reductase activity of the anaerobic archaeon Pyrococcus furiosus, which grows optimally at 100[degrees]C by a metabolism that produces H[sub 2]S if S[sup 0] is present, was found in the cytoplasm. It was purified anaerobically and was shown to be identical to the hydrogenase that had been previously purified from this organism. Both S[sup 0] and polysulfide served as substrates for H[sub 2]S production, and the S[sub 0] reduction activity but not the H[sub 2]-oxidation activity was enhanced by the redox protein rubredoxin. The H[sub 2]-oxidizing and S[sup 0]-reduction activities of the enzyme also showed different responses to pH, temperature, and inhibitors. This bifunctional [open quotes]sulfhydrogenase[close quotes] enzyme can, therefore, dispose of the excess reductant generated during fermentation using either protons or polysulfides as the electron acceptor. In addition, purified hydrogenases from both hyperthermophilic and mesophilic representatives of the archaeal and bacterial domains were shown to reduce S[sup 0] to H[sub 2]S. It is suggested that the function of some form of ancestral hydrogenase was S[sup 0] reduction rather than, or in addition, to the reduction of protons. 33 refs., 4 figs., 2 tabs.

  13. Observation of terahertz vibrations in Pyrococcus furiosus rubredoxin via impulsive coherent vibrational spectroscopy and nuclear resonance vibrational spectroscopy--interpretation by molecular mechanics.

    PubMed

    Tan, Ming-Liang; Bizzarri, Anna Rita; Xiao, Yuming; Cannistraro, Salvatore; Ichiye, Toshiko; Manzoni, Cristian; Cerullo, Giulio; Adams, Michael W W; Jenney, Francis E; Cramer, Stephen P

    2007-03-01

    We have used impulsive coherent vibrational spectroscopy (ICVS) to study the Fe(S-Cys)(4) site in oxidized rubredoxin (Rd) from Pyrococcus furiosus (Pf). In this experiment, a 15 fs visible laser pulse is used to coherently pump the sample to an excited electronic state, and a second <10 fs pulse is used to probe the change in transmission as a function of the time delay. PfRd was observed to relax to the ground state by a single exponential decay with time constants of approximately 255-275 fs. Superimposed on this relaxation are oscillations caused by coherent excitation of vibrational modes in both excited and ground electronic states. Fourier transformation reveals the frequencies of these modes. The strongest ICV mode with 570 nm excitation is the symmetric Fe-S stretching mode near 310 cm(-1), compared to 313 cm(-1) in the low temperature resonance Raman. If the rubredoxin is pumped at 520 nm, a set of strong bands occurs between 20 and 110 cm(-1). Finally, there is a mode at approximately 500 cm(-1) which is similar to features near 508 cm(-1) in blue Cu proteins that have been attributed to excited state vibrations. Normal mode analysis using 488 protein atoms and 558 waters gave calculated spectra that are in good agreement with previous nuclear resonance vibrational spectra (NRVS) results. The lowest frequency normal modes are identified as collective motions of the entire protein or large segments of polypeptide. Motion in these modes may affect the polar environment of the redox site and thus tune the electron transfer functions in rubredoxins.

  14. DNA polymerase hybrids derived from the family-B enzymes of Pyrococcus furiosus and Thermococcus kodakarensis: improving performance in the polymerase chain reaction

    PubMed Central

    Elshawadfy, Ashraf M.; Keith, Brian J.; Ee Ooi, H'Ng; Kinsman, Thomas; Heslop, Pauline; Connolly, Bernard A.

    2014-01-01

    The polymerase chain reaction (PCR) is widely applied across the biosciences, with archaeal Family-B DNA polymerases being preferred, due to their high thermostability and fidelity. The enzyme from Pyrococcus furiosus (Pfu-Pol) is more frequently used than the similar protein from Thermococcus kodakarensis (Tkod-Pol), despite the latter having better PCR performance. Here the two polymerases have been comprehensively compared, confirming that Tkod-Pol: (1) extends primer-templates more rapidly; (2) has higher processivity; (3) demonstrates superior performance in normal and real time PCR. However, Tkod-Pol is less thermostable than Pfu-Pol and both enzymes have equal fidelities. To understand the favorable properties of Tkod-Pol, hybrid proteins have been prepared. Single, double and triple mutations were used to site arginines, present at the “forked-point” (the junction of the exonuclease and polymerase channels) of Tkod-Pol, at the corresponding locations in Pfu-Pol, slightly improving PCR performance. The Pfu-Pol thumb domain, responsible for double-stranded DNA binding, has been entirely replaced with that from Tkod-Pol, again giving better PCR properties. Combining the “forked-point” and thumb swap mutations resulted in a marked increase in PCR capability, maintenance of high fidelity and retention of the superior thermostability associated with Pfu-Pol. However, even the arginine/thumb swap mutant falls short of Tkod-Pol in PCR, suggesting further improvement within the Pfu-Pol framework is attainable. The significance of this work is the observation that improvements in PCR performance are easily attainable by blending elements from closely related archaeal polymerases, an approach that may, in future, be extended by using more polymerases from these organisms. PMID:24904539

  15. TrmBL2 from Pyrococcus furiosus Interacts Both with Double-Stranded and Single-Stranded DNA

    PubMed Central

    Wierer, Sebastian; Daldrop, Peter; Ud Din Ahmad, Misbha; Boos, Winfried; Drescher, Malte; Welte, Wolfram; Seidel, Ralf

    2016-01-01

    In many hyperthermophilic archaea the DNA binding protein TrmBL2 or one of its homologues is abundantly expressed. TrmBL2 is thought to play a significant role in modulating the chromatin architecture in combination with the archaeal histone proteins and Alba. However, its precise physiological role is poorly understood. It has been previously shown that upon binding TrmBL2 covers double-stranded DNA, which leads to the formation of a thick and fibrous filament. Here we investigated the filament formation process as well as the stabilization of DNA by TrmBL2 from Pyroccocus furiosus in detail. We used magnetic tweezers that allow to monitor changes of the DNA mechanical properties upon TrmBL2 binding on the single-molecule level. Extended filaments formed in a cooperative manner and were considerably stiffer than bare double-stranded DNA. Unlike Alba, TrmBL2 did not form DNA cross-bridges. The protein was found to bind double- and single-stranded DNA with similar affinities. In mechanical disruption experiments of DNA hairpins this led to stabilization of both, the double- (before disruption) and the single-stranded (after disruption) DNA forms. Combined, these findings suggest that the biological function of TrmBL2 is not limited to modulating genome architecture and acting as a global repressor but that the protein acts additionally as a stabilizer of DNA secondary structure. PMID:27214207

  16. TrmBL2 from Pyrococcus furiosus Interacts Both with Double-Stranded and Single-Stranded DNA.

    PubMed

    Wierer, Sebastian; Daldrop, Peter; Ud Din Ahmad, Misbha; Boos, Winfried; Drescher, Malte; Welte, Wolfram; Seidel, Ralf

    2016-01-01

    In many hyperthermophilic archaea the DNA binding protein TrmBL2 or one of its homologues is abundantly expressed. TrmBL2 is thought to play a significant role in modulating the chromatin architecture in combination with the archaeal histone proteins and Alba. However, its precise physiological role is poorly understood. It has been previously shown that upon binding TrmBL2 covers double-stranded DNA, which leads to the formation of a thick and fibrous filament. Here we investigated the filament formation process as well as the stabilization of DNA by TrmBL2 from Pyroccocus furiosus in detail. We used magnetic tweezers that allow to monitor changes of the DNA mechanical properties upon TrmBL2 binding on the single-molecule level. Extended filaments formed in a cooperative manner and were considerably stiffer than bare double-stranded DNA. Unlike Alba, TrmBL2 did not form DNA cross-bridges. The protein was found to bind double- and single-stranded DNA with similar affinities. In mechanical disruption experiments of DNA hairpins this led to stabilization of both, the double- (before disruption) and the single-stranded (after disruption) DNA forms. Combined, these findings suggest that the biological function of TrmBL2 is not limited to modulating genome architecture and acting as a global repressor but that the protein acts additionally as a stabilizer of DNA secondary structure.

  17. Characterization of an extremely thermostable but cold-adaptive β-galactosidase from the hyperthermophilic archaeon Pyrococcus furiosus for use as a recombinant aggregation for batch lactose degradation at high temperature.

    PubMed

    Dong, Qing; Yan, Xufan; Zheng, Minhui; Yang, Ziwen

    2014-06-01

    β-Galactosidase (lactase), which catalyzes the hydrolysis of lactose into glucose and galactose, is one of the most important enzymes used in dairy processing. In this study, a gene that encoded an extremely thermostable β-galactosidase from Pyrococcus furiosus (Pflactase) was cloned and expressed in Escherichia coli BL21. The recombinant enzyme was purified by heat treatment and Ni-NTA affinity chromatography. The enzyme displayed optimal activity at 90°C and pH 7.0 in phosphate buffer. The specific activity of the recombinant enzyme on o-nitrophenyl-β-d-galactopyranoside was 10.2 U/mg at 0°C and 130.0dU/mg at 90°C. The half-lives of the enzyme were 31423.4, 8168.3, 4017.7, 547.4, 309.6, and 203.5 min at 70°C, 80°C, 85°C, 90°C, 95°C, and 100°C, respectively. The recombinant enzyme exhibited both β-galactosidase and β-glucosidase activity. The active inclusion bodies of β-galactosidase were easily isolated by nonionic detergent treatment and directly used for lactose conversion in a repetitive batch mode. More than 54% (90°C) or 88% (10°C) of the original enzyme activity was retained after 10 conversion cycles under optimum conditions. These results suggest that the recombinant thermostable β-galactosidase may be suitable for the hydrolysis of lactose in milk processing.

  18. Dynamics of the [4Fe-4S] Cluster in Pyrococcus furiosus D14C Ferredoxin via Nuclear Resonance Vibrational and Resonance Raman Spectroscopies, Force Field Simulations, and Density Functional Theory Calculations

    PubMed Central

    Mitra, Devrani; Pelmenschikov, Vladimir; Guo, Yisong; Case, David A.; Wang, Hongxin; Dong, Weibing; Tan, Ming-Liang; Ichiye, Toshiko; Jenney, Francis E.; Adams, Michael W. W.; Yoda, Yoshitaka; Zhao, Jiyong; Cramer, Stephen P.

    2011-01-01

    We have used 57Fe nuclear resonance vibrational spectroscopy (NRVS) to study oxidized and reduced forms of the [4Fe-4S] cluster in the D14C variant ferredoxin from Pyrococcus furiosus (Pf D14C Fd). To assist the normal mode assignments, we recorded the NRVS of D14C ferredoxin samples with 36S substituted into the [4Fe-4S] cluster bridging sulfide positions, and a model compound without ligand side chains: (Ph4P)2[Fe4S4Cl4]. Several distinct regions of NRVS intensity are identified, ranging from `protein' and torsional modes below 100 cm−1, through bending and breathing modes near 150 cm−1, to strong bands from Fe-S stretching modes between 250 cm−1 and ~400 cm−1. The oxidized ferredoxin samples were also investigated by resonance Raman (RR) spectroscopy. We found good agreement between NRVS and RR frequencies, but because of different selection rules, the intensities vary dramatically between the two types of spectra. The 57Fe partial vibrational densities of states (PVDOS) for the oxidized samples were interpreted by normal mode analysis with optimization of Urey-Bradley force fields for local models of the [4Fe-4S] clusters. Full protein model calculations were also conducted using a supplemented CHARMM force field, and these calculations revealed low frequency modes that may be relevant to electron transfer with Pf Fd partners. Density functional theory (DFT) calculations complemented these empirical analyses, and DFT was used to estimate the reorganization energy associated with the [Fe4S4]2+/1+ redox cycle. Overall, the NRVS technique demonstrates great promise for the observation and quantitative interpretation of the dynamical properties of Fe-S proteins. PMID:21500788

  19. Crystal structure of an intracellular protease from Pyrococcus horikoshii at 2-Å resolution

    PubMed Central

    Du, Xinlin; Choi, In-Geol; Kim, Rosalind; Wang, Weiru; Jancarik, Jaru; Yokota, Hisao; Kim, Sung-Hou

    2000-01-01

    The intracellular protease from Pyrococcus horikoshii (PH1704) and PfpI from Pyrococcus furiosus are members of a class of intracellular proteases that have no sequence homology to any other known protease family. We report the crystal structure of PH1704 at 2.0-Å resolution. The protease is tentatively identified as a cysteine protease based on the presence of cysteine (residue 100) in a nucleophile elbow motif. In the crystal, PH1704 forms a hexameric ring structure, and the active sites are formed at the interfaces between three pairs of monomers. PMID:11114201

  20. Identification of a glycolytic regulon in the archaea Pyrococcus and Thermococcus.

    PubMed

    van de Werken, Harmen J G; Verhees, Corné H; Akerboom, Jasper; de Vos, Willem M; van der Oost, John

    2006-07-01

    The glycolytic pathway of the hyperthermophilic archaea that belong to the order Thermococcales (Pyrococcus, Thermococcus and Palaeococcus) differs significantly from the canonical Embden-Meyerhof pathway in bacteria and eukarya. This archaeal glycolysis variant consists of several novel enzymes, some of which catalyze unique conversions. Moreover, the enzymes appear not to be regulated allosterically, but rather at transcriptional level. To elucidate details of the gene expression control, the transcription initiation sites of the glycolytic genes in Pyrococcus furiosus have been mapped by primer extension analysis and the obtained promoter sequences have been compared with upstream regions of non-glycolytic genes. Apart from consensus sequences for the general transcription factors (TATA-box and BRE) this analysis revealed the presence of a potential transcription factor binding site (TATCAC-N(5)-GTGATA) in glycolytic and starch utilizing promoters of P. furiosus and several thermococcal species. The absence of this inverted repeat in Pyrococcus abyssi and Pyrococcus horikoshii probably reflects that their reduced catabolic capacity does not require this regulatory system. Moreover, this phyletic pattern revealed a TrmB-like regulator (PF0124 and TK1769) which may be involved in recognizing the repeat. This Thermococcales glycolytic regulon, with more than 20 genes, is the largest regulon that has yet been described for Archaea.

  1. Pyrococcus horikoshii sp. nov., a hyperthermophilic archaeon isolated from a hydrothermal vent at the Okinawa Trough.

    PubMed

    González, J M; Masuchi, Y; Robb, F T; Ammerman, J W; Maeder, D L; Yanagibayashi, M; Tamaoka, J; Kato, C

    1998-05-01

    A hyperthermophilic, anaerobic archaeon was isolated from hydrothermal fluid samples obtained at the Okinawa Trough vents in the NE Pacific Ocean, at a depth of 1395m. The strain is obligately heterotrophic, and utilizes complex proteinaceous media (peptone, tryptone, or yeast extract), or a 21-amino-acid mixture supplemented with vitamins, as growth substrates. Sulfur greatly enhances growth. The cells are irregular cocci with a tuft of flagella, growing optimally at 98 degrees C (maximum growth temperature 102 degrees C), but capable of prolonged survival at 105 degrees C. Optimum growth was at pH 7 (range 5-8) and NaCl concentration 2.4% (range 1%-5%). Tryptophan was required for growth, in contrast to the closely related strains Pyrococcus furiosus and P. abyssi. Thin sections of the cell, viewed by transmission electron microscopy, revealed a periplasmic space similar in appearance to the envelope of P. furiosus. The predominant cell membrane component was tetraether lipid, with minor amounts of diether lipids. Treatment of the cells by mild osmotic shock released an extract that contained a Zn(2+)-dependent alkaline phosphatase. Phylogenetic analysis of the sequences encoding 16S rRNA and glutamate dehydrogenase places the isolate with certainty within the genus Pyrococcus although there is relatively low DNA-DNA hybridization (< 63%) with described species of this genus. Based on the reported results, we propose a new species, to be named Pyrococcus horikoshii sp.nov.

  2. Role of Polysulfides in Reduction of Elemental Sulfur by the Hyperthermophilic Archaebacterium Pyrococcus furiosus†

    PubMed Central

    Blumentals, I. I.; Itoh, M.; Olson, G. J.; Kelly, R. M.

    1990-01-01

    Polysulfides formed through the breakdown of elemental sulfur or other sulfur compounds were found to be reduced to H2S by the hyperthermophilic archaebacterium Pyrococcus furiosus during growth. Metabolism of polysulfides by the organism was dissimilatory, as no incorporation of 35S-labeled elemental sulfur was detected. However, [35S]cysteine and [35S]methionine were incorporated into cellular protein. Contact between the organism and elemental sulfur is not necessary for metabolism. The sulfide generated from metabolic reduction of polysulfides dissociates to a strong nucleophile, HS−, which in turn opens up the S8 elemental sulfur ring. In addition to H2S, P. furiosus cultures produced methyl mercaptan in a growth-associated fashion. PMID:16348181

  3. Molybdenum Incorporation in Tungsten Aldehyde Oxidoreductase Enzymes from Pyrococcus furiosus▿ †

    PubMed Central

    Sevcenco, Ana-Maria; Bevers, Loes E.; Pinkse, Martijn W. H.; Krijger, Gerard C.; Wolterbeek, Hubert T.; Verhaert, Peter D. E. M.; Hagen, Wilfred R.; Hagedoorn, Peter-Leon

    2010-01-01

    The hyperthermophilic archaeon Pyrococcus furiosus expresses five aldehyde oxidoreductase (AOR) enzymes, all containing a tungsto-bispterin cofactor. The growth of this organism is fully dependent on the presence of tungsten in the growth medium. Previous studies have suggested that molybdenum is not incorporated in the active site of these enzymes. Application of the radioisotope 99Mo in metal isotope native radioautography in gel electrophoresis (MIRAGE) technology to P. furiosus shows that molybdenum can in fact be incorporated in all five AOR enzymes. Mo(V) signals characteristic for molybdopterin were observed in formaldehyde oxidoreductase (FOR) in electron paramagnetic resonance (EPR)-monitored redox titrations. Our finding that the aldehyde oxidation activity of FOR and WOR5 (W-containing oxidoreductase 5) correlates only with the residual tungsten content suggests that the Mo-containing AORs are most likely inactive. An observed W/Mo antagonism is indicative of tungstate-dependent negative feedback of the expression of the tungstate/molybdate ABC transporter. An intracellular selection mechanism for tungstate and molybdate processing has to be present, since tungsten was found to be preferentially incorporated into the AORs even under conditions with comparable intracellular concentrations of tungstate and molybdate. Under the employed growth conditions of starch as the main carbon source in a rich medium, no tungsten- and/or molybdenum-associated proteins are detected in P. furiosus other than the high-affinity transporter, the proteins of the metallopterin insertion machinery, and the five W-AORs. PMID:20562313

  4. Pyrococcus furiosus-immobilized anodized tubular titania cathode in a hydrogen production system

    NASA Astrophysics Data System (ADS)

    Yoon, Jaekyung; Bae, Sanghyun; Shim, Eunjung; Joo, Hyunku

    Anodized tubular titania (TiO 2) electrodes (ATTEs) are prepared and used as both the photoanode and the cathode substrate in a photoelectrochemical system designed to split water into hydrogen with the assistance of an enzyme and an external bias of 1.5 V. In particular, the ATTE used as the cathode substrate for the immobilization of the enzyme is prepared by two methods-adsorption and crosslinking. Results show that the optimized amount of enzyme is 10.98 units for the slurried enzyme, 3.66 units for the adsorbed one and 7.32 units for the crosslinked one, and the corresponding hydrogen evolution rates are 33.04, 148.58 and 234.88 μmol h -1, respectively. The immobilized enzyme, specifically the chemically crosslinked one, seems to be much superior to the slurried enzyme, due to the enhanced charge-transfer process that is caused by the lower electrical resistance between the enzyme and the ATTE. This results in a greater number of accepted electrons and a larger amount of enzymes able to deal with the electrons.

  5. Evaluation of sulfur-reducing microorganisms for organic desulfurization. [Pyrococcus furiosus

    SciTech Connect

    Miller, K.W.

    1991-01-01

    Because of substantial portion of the sulfur in Illinois coal is organic, microbial desulfurization of sulfidic and thiophenic functionalities could hold great potential for completing pyritic sulfur removal. We are testing the hypothesis that organic sulfur can be reductively removed as H{sub 2}S through the activities of anaerobic microorganisms. Our objectives for this year include the following: (1) To obtain cultures that will reductively desulfurize thiophenic model compounds. In addition to crude oil enrichments begun last year, we sampled municipal sewage sludge. (2) To continue to work toward optimizing the activity of the DBDS-reducing cultures obtained during the previous year. (3) To expand coal desulfurization work to include other coals including Illinois Basin Coal 101 and a North Dakota lignite, which might be more susceptible to the dibenzyldisulfide reducing cultures due to its lower rank. (4) To address the problem of sulfide sorption, by investigating the sorption capacity of coals in addition to Illinois Basin Coal 108.

  6. Features for instantaneous emissions of low-level infrared signals of glucokinase enzyme from Pyrococcus furiosus.

    PubMed

    Torres, Sergio; Mella, Héctor; Reyes, Claudio; Meza, Pablo; Gallardo, Maria J; Staforelli, Juan P

    2015-03-10

    A noncontact infrared (IR) imaging-based methodology and signal recovery tools are applied on an enzyme reaction as a test target. The method is implemented by a long-wave (8-12 μm) IR microbolometer imaging array and a germanium-based IR optical vision. The reaction is carried out by the glucokinase, which produces a rapid exothermal release of energy that is weak, and, even worse, the IR video captured by the uncooled microbolometer detector is affected by spatial and temporal noise with specific complexities. Hitherto, IR-based signal recovery tools have worked with a standard acquisition frequency, which is clearly beyond the time scale of a real scenario. The implications of this (and similar) rapid reactions motivate the designs of a signal recovery method using prior information of the processes to extract and quantify the spontaneity of the enzymatic reaction in a three-dimensional (space and time) single and noncontact online measurement. PMID:25968383

  7. Regulation and Mechanism of Action of the Small Heat Shock Protein from the Hyperthermophilic Archaeon Pyrococcus furiosus†

    PubMed Central

    Laksanalamai, Pongpan; Maeder, Dennis L.; Robb, Frank T.

    2001-01-01

    The small heat shock protein (sHSP) from the hyperthermophile Pyrococcus furiosus was specifically induced at the level of transcription by heat shock at 105°C. The gene encoding this protein was cloned and overexpressed in Escherichia coli. The recombinant sHSP prevented the majority of E. coli proteins from aggregating in vitro for up to 40 min at 105°C. The sHSP also prevented bovine glutamate dehydrogenase from aggregating at 56°C. Survivability of E. coli overexpressing the sHSP was enhanced approximately sixfold during exposure to 50°C for 2 h compared with the control culture, which did not express the sHSP. Apparently, the sHSP confers a survival advantage on mesophilic bacteria by preventing protein aggregation at supraoptimal temperatures. PMID:11489874

  8. Highly thermostable RadA protein from the archaeon Pyrococcus woesei enhances specificity of simplex and multiplex PCR assays.

    PubMed

    Stefanska, Aleksandra; Gaffke, Lidia; Kaczorowska, Anna-Karina; Plotka, Magdalena; Dabrowski, Slawomir; Kaczorowski, Tadeusz

    2016-05-01

    The radA gene of the hyperthermophilic archaeon Pyrococcus woesei (Thermococcales) was cloned and overexpressed in Escherichia coli. The 1050-bp gene codes for a 349-amino-acid polypeptide with an M r of 38,397 which shows 100 % positional amino acid identity to Pyrococcus furiosus RadA and 27.1 % to the E. coli RecA protein. Recombinant RadA was overproduced in Escherichia coli as a His-tagged fusion protein and purified to electrophoretic homogeneity using a simple procedure consisting of ammonium sulfate precipitation and metal-affinity chromatography. In solution RadA exists as an undecamer (11-mer). The protein binds both to ssDNA and dsDNA. RadA has been found to be highly thermostable, it remains almost unaffected by a 4-h incubation at 94 °C. The addition of the RadA protein to either simplex or multiplex PCR assays, significantly improves the specificity of DNA amplification by eliminating non-specific products. Among applications tested the RadA protein proved to be useful in allelic discrimination assay of HADHA gene associated with long-chain 3-hydroxylacyl-CoA dehydrogenase deficiency that in infancy may lead to hypotonia, serious heart and liver problems and even sudden death.

  9. Improving the Catalytic Activity of Hyperthermophilic Pyrococcus horikoshii Prolidase for Detoxification of Organophosphorus Nerve Agents over a Broad Range of Temperatures

    PubMed Central

    Theriot, Casey M.; Semcer, Rebecca L.; Shah, Saumil S.; Grunden, Amy M.

    2011-01-01

    Prolidases hydrolyze Xaa-Pro dipeptides and can also cleave the P-F and P-O bonds found in organophosphorus (OP) compounds, including the nerve agents soman and sarin. Ph1prol (PH0974) has previously been isolated and characterized from Pyrococcus horikoshii and was shown to have higher catalytic activity over a broader pH range, higher affinity for metal, and increased thermostability compared to P. furiosus prolidase, Pfprol (PF1343). To obtain a better enzyme for OP nerve agent decontamination and to investigate the structural factors that may influence protein thermostability and thermoactivity, randomly mutated Ph1prol enzymes were prepared. Four Ph1prol mutants (A195T/G306S-, Y301C/K342N-, E127G/E252D-, and E36V-Ph1prol) were isolated which had greater thermostability and improved activity over a broader range of temperatures against Xaa-Pro dipeptides and OP nerve agents compared to wild type Pyrococcus prolidases. PMID:22162664

  10. Crystal Structural and Functional Analysis of the Putative Dipeptidase from Pyrococcus horikoshii OT3.

    PubMed

    Jeyakanthan, Jeyaraman; Takada, Katsumi; Sawano, Masahide; Ogasahara, Kyoko; Mizutani, Hisashi; Kunishima, Naoki; Yokoyama, Shigeyuki; Yutani, Katsuhide

    2009-01-01

    The crystal structure of a putative dipeptidase (Phdpd) from Pyrococcus horikoshii OT3 was solved using X-ray data at 2.4 A resolution. The protein is folded into two distinct entities. The N-terminal domain consists of the general topology of the alpha/beta fold, and the C-terminal domain consists of five long mixed strands, four helices, and two 3(10) helices. The structure of Phdpd is quite similar to reported structures of prolidases from P. furiosus (Zn-Pfprol) and P. horikoshii (Zn-Phdpd), where Zn ions are observed in the active site resulting in an inactive form. However, Phdpd did not contain metals in the crystal structure and showed prolidase activity in the absence of additional Co ions, whereas the specific activities increased by 5 times in the presence of a sufficient concentration (1.2 mM) of Co ions. The substrate specificities (X-Pro) of Phdpd were broad compared with those of Zn-Phdpd in the presence of Co ions, whose relative activities are 10% or less for substrates other than Met-Pro, which is the most favorable substrate. The binding constants of Zn-Phdpd with three metals (Zn, Co, and Mn) were higher than those of Phdpd and that with Zn was higher by greater than 2 orders, which were determined by DSC experiments. From the structural comparison of both forms and the above experimental results, it could be elucidated why the protein with Zn(2+) ions is inactive.

  11. Impact of Substrate Glycoside Linkage and Elemental Sulfur on Bioenergetics of and Hydrogen Production by the Hyperthermophilic Archaeon Pyrococcus furiosus▿ †

    PubMed Central

    Chou, Chung-Jung; Shockley, Keith R.; Conners, Shannon B.; Lewis, Derrick L.; Comfort, Donald A.; Adams, Michael W. W.; Kelly, Robert M.

    2007-01-01

    Glycoside linkage (cellobiose versus maltose) dramatically influenced bioenergetics to different extents and by different mechanisms in the hyperthermophilic archaeon Pyrococcus furiosus when it was grown in continuous culture at a dilution rate of 0.45 h−1 at 90°C. In the absence of S0, cellobiose-grown cells generated twice as much protein and had 50%-higher specific H2 generation rates than maltose-grown cultures. Addition of S0 to maltose-grown cultures boosted cell protein production fourfold and shifted gas production completely from H2 to H2S. In contrast, the presence of S0 in cellobiose-grown cells caused only a 1.3-fold increase in protein production and an incomplete shift from H2 to H2S production, with 2.5 times more H2 than H2S formed. Transcriptional response analysis revealed that many genes and operons known to be involved in α- or β-glucan uptake and processing were up-regulated in an S0-independent manner. Most differentially transcribed open reading frames (ORFs) responding to S0 in cellobiose-grown cells also responded to S0 in maltose-grown cells; these ORFs included ORFs encoding a membrane-bound oxidoreductase complex (MBX) and two hypothetical proteins (PF2025 and PF2026). However, additional genes (242 genes; 108 genes were up-regulated and 134 genes were down-regulated) were differentially transcribed when S0 was present in the medium of maltose-grown cells, indicating that there were different cellular responses to the two sugars. These results indicate that carbohydrate characteristics (e.g., glycoside linkage) have a major impact on S0 metabolism and hydrogen production in P. furiosus. Furthermore, such issues need to be considered in designing and implementing metabolic strategies for production of biofuel by fermentative anaerobes. PMID:17827328

  12. The three-dimensional structure of TrmB, a transcriptional regulator of dual function in the hyperthermophilic archaeon Pyrococcus furiosus in complex with sucrose.

    PubMed

    Krug, Michael; Lee, Sung-Jae; Boos, Winfried; Diederichs, Kay; Welte, Wolfram

    2013-06-01

    TrmB is a repressor that binds maltose, maltotriose, and sucrose, as well as other α-glucosides. It recognizes two different operator sequences controlling the TM (Trehalose/Maltose) and the MD (Maltodextrin) operon encoding the respective ABC transporters and sugar-degrading enzymes. Binding of maltose to TrmB abrogates repression of the TM operon but maintains the repression of the MD operon. On the other hand, binding of sucrose abrogates repression of the MD operon but maintains repression of the TM operon. The three-dimensional structure of TrmB in complex with sucrose was solved and refined to a resolution of 3.0 Å. The structure shows the N-terminal DNA binding domain containing a winged-helix-turn-helix (wHTH) domain followed by an amphipathic helix with a coiled-coil motif. The latter promotes dimerization and places the symmetry mates of the putative recognition helix in the wHTH motif about 30 Å apart suggesting a canonical binding to two successive major grooves of duplex palindromic DNA. This suggests that the structure resembles the conformation of TrmB recognizing the pseudopalindromic TM promoter but not the conformation recognizing the nonpalindromic MD promoter.

  13. The three-dimensional structure of TrmB, a transcriptional regulator of dual function in the hyperthermophilic archaeon Pyrococcus furiosus in complex with sucrose

    PubMed Central

    Krug, Michael; Lee, Sung-Jae; Boos, Winfried; Diederichs, Kay; Welte, Wolfram

    2013-01-01

    TrmB is a repressor that binds maltose, maltotriose, and sucrose, as well as other α-glucosides. It recognizes two different operator sequences controlling the TM (Trehalose/Maltose) and the MD (Maltodextrin) operon encoding the respective ABC transporters and sugar-degrading enzymes. Binding of maltose to TrmB abrogates repression of the TM operon but maintains the repression of the MD operon. On the other hand, binding of sucrose abrogates repression of the MD operon but maintains repression of the TM operon. The three-dimensional structure of TrmB in complex with sucrose was solved and refined to a resolution of 3.0 Å. The structure shows the N-terminal DNA binding domain containing a winged-helix-turn-helix (wHTH) domain followed by an amphipathic helix with a coiled-coil motif. The latter promotes dimerization and places the symmetry mates of the putative recognition helix in the wHTH motif about 30 Å apart suggesting a canonical binding to two successive major grooves of duplex palindromic DNA. This suggests that the structure resembles the conformation of TrmB recognizing the pseudopalindromic TM promoter but not the conformation recognizing the nonpalindromic MD promoter. PMID:23576322

  14. Cover preference of the Carolina madtom (Noturus furiosus), an imperiled, endemic southeastern stream fish

    USGS Publications Warehouse

    Midway, S.R.; Aday, D.D.; Kwak, T.J.; Gross, K.

    2010-01-01

    In a laboratory setting, we investigated cover preference of the Carolina madtom (Noturus furiosus), an imperiled, endemic southeastern USA stream fish. Fish were tested individually and given 24 hours to make a selection from four cover options, including rock, leaf pack, mussel shell, and an artificial cover unit. Among 30 trials, Carolina madtom preferred the artificial cover unit, selecting it 63% of the time. Rock was selected 23% of the time, and leaf pack 13%. Mussel shells were not selected during any trial.

  15. Cover preference of the Carolina madtom (Noturus furiosus), an imperiled, indemic southeastern stream fish

    USGS Publications Warehouse

    Midway, S.R.; Aday, D.D.; Kwak, Thomas J.; Gross, K.

    2010-01-01

    In a laboratory setting, we investigated cover preference of the Carolina madtom (Noturus furiosus), an imperiled, endemic southeastern USA stream fish. Fish were tested individually and given 24 hours to make a selection from four cover options, including rock, leaf pack, mussel shell, and an artificial cover unit. Among 30 trials, Carolina madtom preferred the artificial cover unit, selecting it 63% of the time. Rock was selected 23% of the time, and leaf pack 13%. Mussel shells were not selected during any trial.

  16. Crystal structure of an archaeal Ski2p-like protein from Pyrococcus horikoshii OT3.

    PubMed

    Zhang, Xiaodong; Nakashima, Takashi; Kakuta, Yoshimitsu; Yao, Min; Tanaka, Isao; Kimura, Makoto

    2008-01-01

    The Ski complex composed of Ski2p, Ski3p, and Ski8p plays an essential role in the 3' to 5' cytoplasmic mRNA degradation pathway in yeast. Ski2p is a putative RNA helicase, belonging in the DExD/H-box protein families and conserved in eukarya as well as in archaea. The gene product (Ph1280p) from the hyperthermophilic archaeon Pyrococcus horikoshii OT3 shows sequence homology with Ski2p, sharing 22.6% identical amino acids with a central region of Ski2p. In order to gain structural information about the Ski2p-like RNA helicase, we overproduced Ph1280p in Escherichia coli cells, and purified it to apparent homogeneity. Ph1280p exhibits DNA/RNA-dependent ATPase activity with an optimal temperature at approximately 90 degrees C. The crystal structure of Ph1280p has been solved at a resolution of 3.5 A using single-wavelength anomalous dispersion (SAD) and selenomethionyl (Se-Met)-substituted protein. Ph1280p comprises four subdomains; the two N-terminal subdomains (N1 and N2) fold into an RecA-like architecture with the conserved helicase motifs, while the two C-terminal subdomains (C1 and C2) fold into alpha-helical structures containing a winged helix (WH)-fold and helix-hairpin-helix (HhH)-fold, respectively. Although the structure of each of the Ph1280p subdomains can be individually superimposed on the corresponding domains in other helicases, such as the Escherichia coli DNA helicase RecQ, the relative orientation of the helicase and C-terminal subdomains in Ph1280p is significantly different from that of other helicases. This structural feature is implicated in substrate specificity for the Ski2-like helicase and would play a critical role in the 3' to 5' cytoplasmic mRNA degradation in the Ski complex. PMID:18042682

  17. Complete genome sequence of the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1 and comparison with Pyrococcus genomes

    PubMed Central

    Fukui, Toshiaki; Atomi, Haruyuki; Kanai, Tamotsu; Matsumi, Rie; Fujiwara, Shinsuke; Imanaka, Tadayuki

    2005-01-01

    The genus Thermococcus, comprised of sulfur-reducing hyperthermophilic archaea, belongs to the order Thermococcales in Euryarchaeota along with the closely related genus Pyrococcus. The members of Thermococcus are ubiquitously present in natural high-temperature environments, and are therefore considered to play a major role in the ecology and metabolic activity of microbial consortia within hot-water ecosystems. To obtain insight into this important genus, we have determined and annotated the complete 2,088,737-base genome of Thermococcus kodakaraensis strain KOD1, followed by a comparison with the three complete genomes of Pyrococcus spp. A total of 2306 coding DNA sequences (CDSs) have been identified, among which half (1165 CDSs) are annotatable, whereas the functions of 41% (936 CDSs) cannot be predicted from the primary structures. The genome contains seven genes for probable transposases and four virus-related regions. Several proteins within these genetic elements show high similarities to those in Pyrococcus spp., implying the natural occurrence of horizontal gene transfer of such mobile elements among the order Thermococcales. Comparative genomics clarified that 1204 proteins, including those for information processing and basic metabolisms, are shared among T. kodakaraensis and the three Pyrococcus spp. On the other hand, among the set of 689 proteins unique to T. kodakaraensis, there are several intriguing proteins that might be responsible for the specific trait of the genus Thermococcus, such as proteins involved in additional pyruvate oxidation, nucleotide metabolisms, unique or additional metal ion transporters, improved stress response system, and a distinct restriction system. PMID:15710748

  18. Characterization of the archaeal ribonuclease P proteins from Pyrococcus horikoshii OT3.

    PubMed

    Terada, Atsushi; Honda, Takashi; Fukuhara, Hideo; Hada, Kazumasa; Kimura, Makoto

    2006-08-01

    Ribonuclease P (RNase P) is a ribonucleoprotein complex involved in the processing of the 5'-leader sequence of precursor tRNA (pre-tRNA). Our earlier study revealed that RNase P RNA (pRNA) and five proteins (PhoPop5, PhoRpp38, PhoRpp21, PhoRpp29, and PhoRpp30) in the hyperthermophilic archaeon Pyrococcus horikoshii OT3 reconstituted RNase P activity that exhibits enzymatic properties like those of the authentic enzyme. In present study, we investigated involvement of the individual proteins in RNase P activity. Two particles (R-3Ps), in which pRNA was mixed with three proteins, PhoPop5, PhoRpp30, and PhoRpp38 or PhoPop5, PhoRpp30, and PhoRpp21 showed a detectable RNase P activity, and five reconstituted particles (R-4Ps) composed of pRNA and four proteins exhibited RNase P activity, albeit at reduced level compared to that of the reconstituted particle (R-5P) composed of pRNA and five proteins. Time-course analysis of the RNase P activities of R-4Ps indicated that the R-4Ps lacking PhoPop5, PhoRpp21, or PhoRpp30 had virtually reduced activity, while omission of PhoRpp29 or PhoRpp38 had a slight effect on the activity. The results indicate that the proteins contribute to RNase P activity in order of PhoPop5 > PhoRpp30 > PhoRpp21 > PhoRpp29 > PhoRpp38. It was further found that R-4Ps showed a characteristic Mg2+ ion dependency approximately identical to that of R-5P. However, R-4Ps had optimum temperature of around at 55 degrees C which is lower than 70 degrees C for R-5P. Together, it is suggested that the P. horikoshii RNase P proteins are predominantly involved in optimization of the pRNA conformation, though they are individually dispensable for RNase P activity in vitro.

  19. High hydrostatic pressure adaptive strategies in an obligate piezophile Pyrococcus yayanosii

    PubMed Central

    Michoud, Grégoire; Jebbar, Mohamed

    2016-01-01

    Pyrococcus yayanosii CH1, as the first and only obligate piezophilic hyperthermophilic microorganism discovered to date, extends the physical and chemical limits of life on Earth. It was isolated from the Ashadze hydrothermal vent at 4,100 m depth. Multi-omics analyses were performed to study the mechanisms used by the cell to cope with high hydrostatic pressure variations. In silico analyses showed that the P. yayanosii genome is highly adapted to its harsh environment, with a loss of aromatic amino acid biosynthesis pathways and the high constitutive expression of the energy metabolism compared with other non-obligate piezophilic Pyrococcus species. Differential proteomics and transcriptomics analyses identified key hydrostatic pressure-responsive genes involved in translation, chemotaxis, energy metabolism (hydrogenases and formate metabolism) and Clustered Regularly Interspaced Short Palindromic Repeats sequences associated with Cellular apoptosis susceptibility proteins. PMID:27250364

  20. High hydrostatic pressure adaptive strategies in an obligate piezophile Pyrococcus yayanosii

    NASA Astrophysics Data System (ADS)

    Michoud, Grégoire; Jebbar, Mohamed

    2016-06-01

    Pyrococcus yayanosii CH1, as the first and only obligate piezophilic hyperthermophilic microorganism discovered to date, extends the physical and chemical limits of life on Earth. It was isolated from the Ashadze hydrothermal vent at 4,100 m depth. Multi-omics analyses were performed to study the mechanisms used by the cell to cope with high hydrostatic pressure variations. In silico analyses showed that the P. yayanosii genome is highly adapted to its harsh environment, with a loss of aromatic amino acid biosynthesis pathways and the high constitutive expression of the energy metabolism compared with other non-obligate piezophilic Pyrococcus species. Differential proteomics and transcriptomics analyses identified key hydrostatic pressure-responsive genes involved in translation, chemotaxis, energy metabolism (hydrogenases and formate metabolism) and Clustered Regularly Interspaced Short Palindromic Repeats sequences associated with Cellular apoptosis susceptibility proteins.

  1. Aspartate transcarbamylase from the deep-sea hyperthermophilic archaeon Pyrococcus abyssi: genetic organization, structure, and expression in Escherichia coli.

    PubMed

    Purcarea, C; Hervé, G; Ladjimi, M M; Cunin, R

    1997-07-01

    The genes coding for aspartate transcarbamylase (ATCase) in the deep-sea hyperthermophilic archaeon Pyrococcus abyssi were cloned by complementation of a pyrB Escherichia coli mutant. The sequence revealed the existence of a pyrBI operon, coding for a catalytic chain and a regulatory chain, as in Enterobacteriaceae. Comparison of primary sequences of the polypeptides encoded by the pyrB and pyrI genes with those of homologous eubacterial and eukaryotic chains showed a high degree of conservation of the residues which in E. coli ATCase are involved in catalysis and allosteric regulation. The regulatory chain shows more-extensive divergence with respect to that of E. coli and other Enterobacteriaceae than the catalytic chain. Several substitutions suggest the existence in P. abyssi ATCase of additional hydrophobic interactions and ionic bonds which are probably involved in protein stabilization at high temperatures. The catalytic chain presents a secondary structure similar to that of the E. coli enzyme. Modeling of the tridimensional structure of this chain provides a folding close to that of the E. coli protein in spite of several significant differences. Conservation of numerous pairs of residues involved in the interfaces between different chains or subunits in E. coli ATCase suggests that the P. abyssi enzyme has a quaternary structure similar to that of the E. coli enzyme. P. abyssi ATCase expressed in transgenic E. coli cells exhibited reduced cooperativity for aspartate binding and sensitivity to allosteric effectors, as well as a decreased thermostability and barostability, suggesting that in P. abyssi cells this enzyme is further stabilized through its association with other cellular components.

  2. Mechanism of protein splicing of the Pyrococcus abyssi lon protease intein

    SciTech Connect

    O'Brien, Kevin M.; Schufreider, Ann K.; McGill, Melissa A.; O'Brien, Kathryn M.; Reitter, Julie N.; Mills, Kenneth V.

    2010-12-17

    Research highlights: {yields} The Pyrococcus abyssi lon protease intein promotes efficient protein splicing. {yields} Inteins with mutations that interfere with individual steps of splicing do not promote unproductive side reactions. {yields} The intein splices with Lys in place of the highly conserved penultimate His. {yields} The intein is flanked by a Gly-rich region at its C terminus that may increase the efficiency of the third step of splicing, Asn cyclization coupled to peptide bond cleavage. -- Abstract: Protein splicing is a post-translational process by which an intervening polypeptide, the intein, excises itself from the flanking polypeptides, the exteins, coupled to ligation of the exteins. The lon protease of Pyrococcus abyssi (Pab) is interrupted by an intein. When over-expressed as a fusion protein in Escherichia coli, the Pab lon protease intein can promote efficient protein splicing. Mutations that block individual steps of splicing generally do not lead to unproductive side reactions, suggesting that the intein tightly coordinates the splicing process. The intein can splice, although it has Lys in place of the highly conserved penultimate His, and mutants of the intein in the C-terminal region lead to the accumulation of stable branched-ester intermediate.

  3. Outdoor Exhibits

    NASA Technical Reports Server (NTRS)

    1996-01-01

    The National Data Buoy Center (NDBC) at the John C. Stennis Space Center has exhibits located in front of the Visitors Center. These boat-shaped buoys are moored in areas of the ocean that experience hostile environmental conditions. The instruments installed gather information and relay it to the National Weather Service by satellite. Nomad buoys are 20 feet long and weigh 13,900 pounds. They provide information on wind speed and direction, humidity levels, air and sea surface temperature and air pressure. U.S. Coast Guard ships transport buoys to their mooring sites.

  4. Archaeal homologs of eukaryotic methylation guide small nucleolar RNAs: lessons from the Pyrococcus genomes.

    PubMed

    Gaspin, C; Cavaillé, J; Erauso, G; Bachellerie, J P

    2000-04-01

    Ribose methylation is a prevalent type of nucleotide modification in rRNA. Eukaryotic rRNAs display a complex pattern of ribose methylations, amounting to 55 in yeast Saccharomyces cerevisiae and about 100 in vertebrates. Ribose methylations of eukaryotic rRNAs are each guided by a cognate small RNA, belonging to the family of box C/D antisense snoRNAs, through transient formation of a specific base-pairing at the rRNA modification site. In prokaryotes, the pattern of rRNA ribose methylations has been fully characterized in a single species so far, Escherichia coli, which contains only four ribose methylated rRNA nucleotides. However, the hyperthermophile archaeon Sulfolobus solfataricus contains, like eukaryotes, a large number of (yet unmapped) rRNA ribose methylations and homologs of eukaryotic box C/D small nucleolar ribonuclear proteins have been identified in archaeal genomes. We have therefore searched archaeal genomes for potential homologs of eukaryotic methylation guide small nucleolar RNAs, by combining searches for structured motifs with homology searches. We have identified a family of 46 small RNAs, conserved in the genomes of three hyperthermophile Pyrococcus species, which we have experimentally characterized in Pyrococcus abyssi. The Pyrococcus small RNAs, the first reported homologs of methylation guide small nucleolar RNAs in organisms devoid of a nucleus, appear as a paradigm of minimalist box C/D antisense RNAs. They differ from their eukaryotic homologs by their outstanding structural homogeneity, extended consensus box motifs and the quasi-systematic presence of two (instead of one) rRNA antisense elements. Remarkably, for each small RNA the two antisense elements always match rRNA sequences close to each other in rRNA structure, suggesting an important role in rRNA folding. Only a few of the predicted P. abyssi rRNA ribose methylations have been detected so far. Further analysis of these archaeal small RNAs could provide new insights into

  5. The crystal structure of a novel SAM-dependent methyltransferase PH1915 from Pyrococcus horikoshii.

    SciTech Connect

    Sun, W.; Xu, X.; Pavlova, M.; Edwards, A.; Joachimiak, A.; Savchenko, A.; Christendat, D.; Biosciences Division; Univ. of Toronto; Univ. Health Network

    2005-01-01

    The S-adenosyl-L-methionine (SAM)-dependent methyltransferases represent a diverse and biologically important class of enzymes. These enzymes utilize the ubiquitous methyl donor SAM as a cofactor to methylate proteins, small molecules, lipids, and nucleic acids. Here we present the crystal structure of PH1915 from Pyrococcus horikoshii OT3, a predicted SAM-dependent methyltransferase. This protein belongs to the Cluster of Orthologous Group 1092, and the presented crystal structure is the first representative structure of this protein family. Based on sequence and 3D structure analysis, we have made valuable functional insights that will facilitate further studies for characterizing this group of proteins. Specifically, we propose that PH1915 and its orthologs are rRNA- or tRNA-specific methyltransferases.

  6. Cloning, purification, crystallization and preliminary crystallographic analysis of acylphosphatase from Pyrococcus horikoshii OT3.

    PubMed

    Miyazono, Ken Ichi; Kudo, Norio; Tanokura, Masaru

    2004-06-01

    Acylphosphatase is one of the smallest enzymes and catalyzes the hydrolysis of the carboxy-phosphate bond. An extremely thermostable acylphosphatase from a hyperthermophilic archaea, Pyrococcus horikoshii OT3, has been cloned, expressed in Escherichia coli, purified and crystallized using the sitting-drop vapour-diffusion method with potassium/sodium tartrate as the precipitant at pH 5.5. X-ray diffraction data have been collected to a highest resolution of 1.72 angstroms on a synchrotron-radiation source. The crystals belong to space group P3(2)21, with approximate unit-cell parameters a = b = 86.6, c = 75.4 angstroms and two monomers in the asymmetric unit.

  7. Complete Genome Sequence of the Hyperthermophilic Archaeon Pyrococcus sp. Strain ST04, Isolated from a Deep-Sea Hydrothermal Sulfide Chimney on the Juan de Fuca Ridge

    PubMed Central

    Jung, Jong-Hyun; Lee, Ju-Hoon; Holden, James F.; Seo, Dong-Ho; Shin, Hakdong; Kim, Hae-Yeong; Kim, Wooki; Ryu, Sangryeol

    2012-01-01

    Pyrococcus sp. strain ST04 is a hyperthermophilic, anaerobic, and heterotrophic archaeon isolated from a deep-sea hydrothermal sulfide chimney on the Endeavour Segment of the Juan de Fuca Ridge in the northeastern Pacific Ocean. To further understand the distinct characteristics of this archaeon at the genome level (polysaccharide utilization at high temperature and ATP generation by a Na+ gradient), the genome of strain ST04 was completely sequenced and analyzed. Here, we present the complete genome sequence analysis results of Pyrococcus sp. ST04 and report the major findings from the genome annotation, with a focus on its saccharolytic and metabolite production potential. PMID:22843576

  8. Complete genome sequence of the hyperthermophilic archaeon Pyrococcus sp. strain ST04, isolated from a deep-sea hydrothermal sulfide chimney on the Juan de Fuca Ridge.

    PubMed

    Jung, Jong-Hyun; Lee, Ju-Hoon; Holden, James F; Seo, Dong-Ho; Shin, Hakdong; Kim, Hae-Yeong; Kim, Wooki; Ryu, Sangryeol; Park, Cheon-Seok

    2012-08-01

    Pyrococcus sp. strain ST04 is a hyperthermophilic, anaerobic, and heterotrophic archaeon isolated from a deep-sea hydrothermal sulfide chimney on the Endeavour Segment of the Juan de Fuca Ridge in the northeastern Pacific Ocean. To further understand the distinct characteristics of this archaeon at the genome level (polysaccharide utilization at high temperature and ATP generation by a Na(+) gradient), the genome of strain ST04 was completely sequenced and analyzed. Here, we present the complete genome sequence analysis results of Pyrococcus sp. ST04 and report the major findings from the genome annotation, with a focus on its saccharolytic and metabolite production potential.

  9. Complete genome sequence of the hyperthermophilic archaeon Pyrococcus sp. strain ST04, isolated from a deep-sea hydrothermal sulfide chimney on the Juan de Fuca Ridge.

    PubMed

    Jung, Jong-Hyun; Lee, Ju-Hoon; Holden, James F; Seo, Dong-Ho; Shin, Hakdong; Kim, Hae-Yeong; Kim, Wooki; Ryu, Sangryeol; Park, Cheon-Seok

    2012-08-01

    Pyrococcus sp. strain ST04 is a hyperthermophilic, anaerobic, and heterotrophic archaeon isolated from a deep-sea hydrothermal sulfide chimney on the Endeavour Segment of the Juan de Fuca Ridge in the northeastern Pacific Ocean. To further understand the distinct characteristics of this archaeon at the genome level (polysaccharide utilization at high temperature and ATP generation by a Na(+) gradient), the genome of strain ST04 was completely sequenced and analyzed. Here, we present the complete genome sequence analysis results of Pyrococcus sp. ST04 and report the major findings from the genome annotation, with a focus on its saccharolytic and metabolite production potential. PMID:22843576

  10. Identification and Characterization of an Archaeal Kojibiose Catabolic Pathway in the Hyperthermophilic Pyrococcus sp. Strain ST04

    PubMed Central

    Jung, Jong-Hyun; Seo, Dong-Ho; Holden, James F.

    2014-01-01

    A unique gene cluster responsible for kojibiose utilization was identified in the genome of Pyrococcus sp. strain ST04. The proteins it encodes hydrolyze kojibiose, a disaccharide product of glucose caramelization, and form glucose-6-phosphate (G6P) in two steps. Heterologous expression of the kojibiose-related enzymes in Escherichia coli revealed that two genes, Py04_1502 and Py04_1503, encode kojibiose phosphorylase (designated PsKP, for Pyrococcus sp. strain ST04 kojibiose phosphorylase) and β-phosphoglucomutase (PsPGM), respectively. Enzymatic assays show that PsKP hydrolyzes kojibiose to glucose and β-glucose-1-phosphate (β-G1P). The Km values for kojibiose and phosphate were determined to be 2.53 ± 0.21 mM and 1.34 ± 0.04 mM, respectively. PsPGM then converts β-G1P into G6P in the presence of 6 mM MgCl2. Conversion activity from β-G1P to G6P was 46.81 ± 3.66 U/mg, and reverse conversion activity from G6P to β-G1P was 3.51 ± 0.13 U/mg. The proteins are highly thermostable, with optimal temperatures of 90°C for PsKP and 95°C for PsPGM. These results indicate that Pyrococcus sp. strain ST04 converts kojibiose into G6P, a substrate of the glycolytic pathway. This is the first report of a disaccharide utilization pathway via phosphorolysis in hyperthermophilic archaea. PMID:24391053

  11. Preliminary crystallographic analysis of an extremely thermostable glutamate dehydrogenase from the archaeon Pyrococcus woesei.

    PubMed

    Knapp, S; Karshikoff, A; Waldkötter, K; Rüdiger, A; Antranikian, G; Ladenstein, R

    1995-05-01

    The extremely heat-stable glutamate dehydrogenase (GluDH) from Pyrococcus woesei was crystallized by the hanging-drop vapour-diffusion method. Crystals suitable for X-ray crystallographic investigations were obtained using polyethylene glycol (PEG) 4000 and ammonium acetate as precipitating agents. The crystals obtained diffract to a resolution of 2.8 A, have a prismatic shape and grow up to 0.5 mm in their maximal dimension. They belong to the triclinic system (space group P1; a = 90.9, b = 92.8, c = 107.1 A, alpha = 69.1, beta = 80.7, Vgamma = 65.0 degrees ) with a unit-cell volume of 765052 A(3) which accommodates one GluDH hexamer of 276 kDa. The averaged density of the crystal determined by Ficoll-gradient centrifugation is 1.15 g cm(-3), which corresponds to a molecular mass of 255 kDa in the unit cell. A native data set has been collected on an MAR image-plate system using Cu Kalpha radiation. The completeness of the data set in the range 316-3 A is 74%, and contains 64% of the data in the outer shell (3.4-2.8 A), with an average R(merge) value of 9%. Calculation of self-rotation functions revealed the 32 symmetry of the hexamer; 3 non-crystallographic twofold axes were found at a distance of 120 degrees in a plane perpendicular to the non-crystallographic threefold axis. PMID:15299309

  12. Substrate recognition of N,N'-diacetylchitobiose deacetylase from Pyrococcus horikoshii.

    PubMed

    Nakamura, Tsutomu; Yonezawa, Yasushige; Tsuchiya, Yuko; Niiyama, Mayumi; Ida, Kurumi; Oshima, Maki; Morita, Junji; Uegaki, Koichi

    2016-09-01

    Enzymes of carbohydrate esterase (CE) family 14 catalyze hydrolysis of N-acetyl groups at the non-reducing end of the N-acetylglucosamine (GlcNAc) residue of chitooligosaccharides or related compounds. N,N'-diacetylchitobiose deacetylase (Dac) belongs to the CE-14 family and plays a role in the chitinolytic pathway in archaea by deacetylating N,N'-diacetylchitobiose (GlcNAc2), which is the end product of chitinase. In this study, we revealed the structural basis of reaction specificity in CE-14 deacetylases by solving a crystal structure of Dac from Pyrococcus horikoshii (Ph-Dac) in complex with a novel reaction intermediate analog. We developed 2-deoxy-2-methylphosphoramido-d-glucose (MPG) as the analog of the tetrahedral oxyanion intermediate of the monosaccharide substrate GlcNAc. The crystal structure of Ph-Dac in complex with MPG demonstrated that Arg92, Asp115, and His152 side chains interact with hydroxyl groups of the glucose moiety of the non-reducing-end GlcNAc residue. The amino acid residues responsible for recognition of the MPG glucose moiety are spatially conserved in other CE-14 deacetylases. Molecular dynamics simulation of the structure of the Ph-Dac-GlcNAc2 complex indicated that the reducing GlcNAc residue is placed in a large intermolecular cleft and is not involved with specific interactions with the enzyme. This observation was consistent with results indicating that Ph-Dac displayed similar kinetic parameters for both GlcNAc and GlcNAc2. This study provides the structural basis of reaction-site specificity of Dac and related CE-14 enzymes.

  13. Substrate recognition of N,N'-diacetylchitobiose deacetylase from Pyrococcus horikoshii.

    PubMed

    Nakamura, Tsutomu; Yonezawa, Yasushige; Tsuchiya, Yuko; Niiyama, Mayumi; Ida, Kurumi; Oshima, Maki; Morita, Junji; Uegaki, Koichi

    2016-09-01

    Enzymes of carbohydrate esterase (CE) family 14 catalyze hydrolysis of N-acetyl groups at the non-reducing end of the N-acetylglucosamine (GlcNAc) residue of chitooligosaccharides or related compounds. N,N'-diacetylchitobiose deacetylase (Dac) belongs to the CE-14 family and plays a role in the chitinolytic pathway in archaea by deacetylating N,N'-diacetylchitobiose (GlcNAc2), which is the end product of chitinase. In this study, we revealed the structural basis of reaction specificity in CE-14 deacetylases by solving a crystal structure of Dac from Pyrococcus horikoshii (Ph-Dac) in complex with a novel reaction intermediate analog. We developed 2-deoxy-2-methylphosphoramido-d-glucose (MPG) as the analog of the tetrahedral oxyanion intermediate of the monosaccharide substrate GlcNAc. The crystal structure of Ph-Dac in complex with MPG demonstrated that Arg92, Asp115, and His152 side chains interact with hydroxyl groups of the glucose moiety of the non-reducing-end GlcNAc residue. The amino acid residues responsible for recognition of the MPG glucose moiety are spatially conserved in other CE-14 deacetylases. Molecular dynamics simulation of the structure of the Ph-Dac-GlcNAc2 complex indicated that the reducing GlcNAc residue is placed in a large intermolecular cleft and is not involved with specific interactions with the enzyme. This observation was consistent with results indicating that Ph-Dac displayed similar kinetic parameters for both GlcNAc and GlcNAc2. This study provides the structural basis of reaction-site specificity of Dac and related CE-14 enzymes. PMID:27456364

  14. Structural features underlying the selective cleavage of a novel exo-type maltose-forming amylase from Pyrococcus sp. ST04.

    PubMed

    Park, Kwang-Hyun; Jung, Jong-Hyun; Park, Sung-Goo; Lee, Myeong-Eun; Holden, James F; Park, Cheon-Seok; Woo, Eui-Jeon

    2014-06-01

    A novel maltose-forming α-amylase (PSMA) was recently found in the hyperthermophilic archaeon Pyrococcus sp. ST04. This enzyme shows <13% amino-acid sequence identity to other known α-amylases and displays a unique enzymatic property in that it hydrolyzes both α-1,4-glucosidic and α-1,6-glucosidic linkages of substrates, recognizing only maltose units, in an exo-type manner. Here, the crystal structure of PSMA at a resolution of 1.8 Å is reported, showing a tight ring-shaped tetramer with monomers composed of two domains: an N-domain (amino acids 1-341) with a typical GH57 family (β/α)7-barrel fold and a C-domain (amino acids 342-597) composed of α-helical bundles. A small closed cavity observed in proximity to the catalytic residues Glu153 and Asp253 at the domain interface has the appropriate volume and geometry to bind a maltose unit, accounting for the selective exo-type maltose hydrolysis of the enzyme. A narrow gate at the putative subsite +1 formed by residue Phe218 and Phe452 is essential for specific cleavage of glucosidic bonds. The closed cavity at the active site is connected to a short substrate-binding channel that extends to the central hole of the tetramer, exhibiting a geometry that is significantly different from classical maltogenic amylases or β-amylases. The structural features of this novel exo-type maltose-forming α-amylase provide a molecular basis for its unique enzymatic characteristics and for its potential use in industrial applications and protein engineering.

  15. Cloning and sequencing of the gene encoding glutamine synthetase I from the archaeum Pyrococcus woesei: anomalous phylogenies inferred from analysis of archaeal and bacterial glutamine synthetase I sequences.

    PubMed Central

    Tiboni, O; Cammarano, P; Sanangelantoni, A M

    1993-01-01

    The gene glnA encoding glutamine synthetase I (GSI) from the archaeum Pyrococcus woesei was cloned and sequenced with the Sulfolobus solfataricus glnA gene as the probe. An operon reading frame of 448 amino acids was identified within a DNA segment of 1,528 bp. The encoded protein was 49% identical with the GSI of Methanococcus voltae and exhibited conserved regions characteristic of the GSI family. The P. woesei GSI was aligned with available homologs from other archaea (S. solfataricus, M. voltae) and with representative sequences from cyanobacteria, proteobacteria, and gram-positive bacteria. Phylogenetic trees were constructed from both the amino acid and the nucleotide sequence alignments. In accordance with the sequence similarities, archaeal and bacterial sequences did not segregate on a phylogeny. On the basis of sequence signatures, the GSI trees could be subdivided into two ensembles. One encompassed the GSI of cyanobacteria and proteobacteria, but also that of the high-G + C gram-positive bacterium Streptomyces coelicolor (all of which are regulated by the reversible adenylylation of the enzyme subunits); the other embraced the GSI of the three archaea as well as that of the low-G + C gram-positive bacteria (Clostridium acetobutilycum, Bacillus subtilis) and Thermotoga maritima (none of which are regulated by subunit adenylylation). The GSIs of the Thermotoga and the Bacillus-Clostridium lineages shared a direct common ancestor with that of P. woesei and the methanogens and were unrelated to their homologs from cyanobacteria, proteobacteria, and S. coelicolor. The possibility is presented that the GSI gene arose among the archaea and was then laterally transferred from some early methanogen to a Thermotoga-like organism. However, the relationship of the cyanobacterial-proteobacterial GSIs to the Thermotoga GSI and the GSI of low-G+C gram-positive bacteria remains unexplained. PMID:8098326

  16. Identification of a novel amino acid racemase from a hyperthermophilic archaeon Pyrococcus horikoshii OT-3 induced by D-amino acids.

    PubMed

    Kawakami, Ryushi; Ohmori, Taketo; Sakuraba, Haruhiko; Ohshima, Toshihisa

    2015-08-01

    To date, there have been few reports analyzing the amino acid requirement for growth of hyperthermophilic archaea. We here found that the hyperthermophilic archaeon Pyrococcus horikoshii OT-3 requires Thr, Leu, Val, Phe, Tyr, Trp, His and Arg in the medium for growth, and shows slow growth in medium lacking Met or Ile. This largely corresponds to the presence, or absence, of genes related to amino acid biosynthesis in its genome, though there are exceptions. The amino acid requirements were dramatically lost by addition of D-isomers of Met, Leu, Val, allo-Ile, Phe, Tyr, Trp and Arg. Tracer analysis using (14)C-labeled D-Trp showed that D-Trp in the medium was used as a protein component in the cells, suggesting the presence of D-amino acid metabolic enzymes. Pyridoxal 5'-phosphate (PLP)-dependent racemase activity toward Met, Leu and Phe was detected in crude extract of P. horikoshii and was enhanced in cells grown in the medium supplemented with D-amino acids, especially D-allo-Ile. The gene encoding the racemase was narrowed down to one open reading frame on the basis of enzyme purification from P. horikoshii cells, and the recombinant enzyme exhibited PLP-dependent racemase activity toward several amino acids, including Met, Leu and Phe, but not Pro, Asp or Glu. This is the first report showing the presence in a hyperthermophilic archaeon of a PLP-dependent amino acid racemase with broad substrate specificity that is likely responsible for utilization of D-amino acids for growth.

  17. Ethics on Exhibit

    ERIC Educational Resources Information Center

    Vick, Randy M.

    2011-01-01

    This article discusses ethical questions raised by an exhibition of work by an artist with a history of mental illness and the exhibition's relevance to art therapy and “outsider art” discourse on the subject. Considerations for how such an exhibit could be handled had the circumstances included an art therapist and art therapy client are…

  18. Thermostable flap endonuclease from the archaeon, Pyrococcus horikoshii, cleaves the replication fork-like structure endo/exonucleolytically.

    PubMed

    Matsui, E; Kawasaki, S; Ishida, H; Ishikawa, K; Kosugi, Y; Kikuchi, H; Kawarabayashi, Y; Matsui, I

    1999-06-25

    The flap endonuclease gene homologue from the hyperthermophilic archaeon, Pyrococcus horikoshii, was overexpressed in Escherichia coli and purified. The results of gel filtration indicated that this protein was a 41-kDa monomer. P. horikoshii flap endonuclease (phFEN) cleaves replication fork-like substrates (RF) and 5' double-strand flap structures (DF) using both flap endonuclease and 5'-3'-exonuclease activities. The mammalian flap endonuclease (mFEN) is a single-strand flap-specific endonuclease (Harrington, J. J., and Lieber, M. R. (1994) EMBO J. 13, 1235-1246), but the action patterns of phFEN appear to be quite different from those of mFEN at this point. The DF-specific flap endonuclease and 5'-exonuclease activities have not yet been reported. Therefore, this is the first report of the specific endo/exonuclease activities of phFEN. The DF-specific 5'-exonuclease activity degraded the downstream primer of 3' single-flap structure and was 15 times higher than the activities against nicked substrates without 3' flap strand. DF-specific flap endonuclease cleaved the 5' double-flap strand in DF and the lagging strand in RF at the junction portion. Because the RF appears to be the intermediate structure, due to the arrest of the replication fork, the double strand breaks after the arrests of the replication forks are probably caused by phFEN.

  19. Cloning, purification, crystallization and preliminary crystallographic analysis of a penicillin-binding protein homologue from Pyrococcus abyssi

    SciTech Connect

    Delfosse, Vanessa; Hugonnet, Jean-Emmanuel; Sougakoff, Wladimir; Mayer, Claudine

    2005-11-01

    The crystallization of a hypothetical penicillin-binding protein from the archaeon P. abyssi in space group C2 by hanging-drop vapour diffusion is reported. The genome of the hyperthermophilic archaeon Pyrococcus abyssi contains a gene (pab0087) encoding a penicillin-binding protein (PBP) homologue. This sequence consists of 447 residues and shows significant sequence similarity to low-molecular-weight PBPs and class C β-lactamases. The Pab0087 protein was overexpressed, purified and crystallized. Diffraction data from two different crystal forms were collected to 2.7 and 2.0 Å resolution. Both crystals belong to space group C2, with unit-cell parameters a = 160.59, b = 135.74, c = 113.02 Å, β = 117.36° and a = 166.97, b = 131.25, c = 189.39 Å, β = 113.81°, respectively. The asymmetric unit contains four and eight molecules, respectively, with fourfold non-crystallographic symmetry.

  20. Characterization of the PH1704 Protease from Pyrococcus horikoshii OT3 and the Critical Functions of Tyr120

    PubMed Central

    Yu, Lei; Han, Weiwei; Feng, Yan

    2014-01-01

    The PH1704 protease from hyperthermophilic archaean Pyrococcus horikoshii OT3 is a member of DJ-1/ThiJ/PfpI superfamily with diverse functional subclasses. The recombinant PH1704 was efficiently purified and was systematically characterized by a combination of substrate specificity analysis, steady-state kinetics study and molecular docking research. The homogeneous protease was obtained as a presumed dodecamer with molecular weight of ∼240 kDa. Iodoacetamide strongly inhibited the peptidase activity, confirming that Cys100 is a nucleophilic residue. The recombinant protein was identified as both an aminopeptidase and an endopeptidase. Experimental data showed that L-R-amc was the best substrate of PH1704. Structural interaction fingerprint analysis (SIFt) indicated the binding pose of PH1704 and showed that Tyr120 is important in substrate binding. Kinetic parameters Kcat and Kcat/Km of the Y120P mutant with L-R-amc was about 7 and 7.8 times higher than that of the wild type (WT). For the endopeptidase Y120P with AAFR-amc, Kcat and Kcat/Km is 10- and 21- fold higher than that of WT. Experimental data indicate the important functions of Tyr120: involvement in enzyme activity to form a hydrogen bond with Cys100 and as an entrance gate of the substrate with Lys43. The results of this study can be used to investigate the DJ-1/ThiJ/PfpI superfamily. PMID:25192005

  1. Microwave-Assisted Synthesis of Glycoconjugates by Transgalactosylation with Recombinant Thermostable β-Glycosidase from Pyrococcus

    PubMed Central

    Henze, Manja; Merker, Dorothee; Elling, Lothar

    2016-01-01

    The potential of the hyperthermophilic β-glycosidase from Pyrococcus woesei (DSM 3773) for the synthesis of glycosides under microwave irradiation (MWI) at low temperatures was investigated. Transgalactosylation reactions with β-N-acetyl-d-glucosamine as acceptor substrate (GlcNAc-linker-tBoc) under thermal heating (TH, 85 °C) and under MWI at 100 and 300 W resulted in the formation of (Galβ(1,4)GlcNAc-linker-tBoc) as the main product in all reactions. Most importantly, MWI at temperatures far below the temperature optimum of the hyperthermophilic glycosidase led to higher product yields with only minor amounts of side products β(1,6-linked disaccharide and trisaccharides). At high acceptor concentrations (50 mM), transgalactosylation reactions under MWI at 300 W gave similar product yields when compared to TH at 85 °C. In summary, we demonstrate that MWI is useful as a novel experimental set-up for the synthesis of defined galacto-oligosaccharides. In conclusion, glycosylation reactions under MWI at low temperatures have the potential as a general strategy for regioselective glycosylation reactions of hyperthermophilic glycosidases using heat-labile acceptor or donor substrates. PMID:26861292

  2. Purification, crystallization and preliminary crystallographic analysis of the vacuole-type ATPase subunit E from Pyrococcus horikoshii OT3

    SciTech Connect

    Lokanath, Neratur K.; Ukita, Yoko; Sugahara, Mitsuaki; Kunishima, Naoki

    2005-01-01

    The E subunit of vacuole-type ATPase from P. horikoshii OT3 was overexpressed, purified and crystallized. The native crystals diffracted X-rays to 1.85 Å resolution. The vacuole-type ATPases in eukaryotic cells translocate protons across various biological membranes including the vacuolar membrane by consuming ATP molecules. The E subunit of the multisubunit complex V-ATPase from Pyrococcus horikoshii OT3, which has a molecular weight of 22.88 kDa, has been cloned, overexpressed in Escherichia coli, purified and crystallized by the microbatch method using PEG 4000 as a precipitant at 296 K. A data set to 1.85 Å resolution with 98.8% completeness and an R{sub merge} of 6.5% was collected from a single flash-cooled crystal using synchrotron radiation. The crystal belonged to the orthorhombic space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 52.196, b = 55.317, c = 77.481 Å, and is most likely to contain one molecule per asymmetric unit.

  3. A Teaching Aids Exhibition.

    ERIC Educational Resources Information Center

    Mahanja, Salah

    1985-01-01

    Describes an exhibition for the benefit of teachers of English in Arab Primary Schools, which was prepared by third-year students at the Teachers College for Arab Teachers. The exhibition included games, songs, audiovisual aids, crossword puzzles, vocabulary, spelling booklets, preposition aids, and worksheet and lesson planning aids. (SED)

  4. Molecular dynamics simulations of the Nip7 proteins from the marine deep- and shallow-water Pyrococcus species

    PubMed Central

    2014-01-01

    Background The identification of the mechanisms of adaptation of protein structures to extreme environmental conditions is a challenging task of structural biology. We performed molecular dynamics (MD) simulations of the Nip7 protein involved in RNA processing from the shallow-water (P. furiosus) and the deep-water (P. abyssi) marine hyperthermophylic archaea at different temperatures (300 and 373 K) and pressures (0.1, 50 and 100 MPa). The aim was to disclose similarities and differences between the deep- and shallow-sea protein models at different temperatures and pressures. Results The current results demonstrate that the 3D models of the two proteins at all the examined values of pressures and temperatures are compact, stable and similar to the known crystal structure of the P. abyssi Nip7. The structural deviations and fluctuations in the polypeptide chain during the MD simulations were the most pronounced in the loop regions, their magnitude being larger for the C-terminal domain in both proteins. A number of highly mobile segments the protein globule presumably involved in protein-protein interactions were identified. Regions of the polypeptide chain with significant difference in conformational dynamics between the deep- and shallow-water proteins were identified. Conclusions The results of our analysis demonstrated that in the examined ranges of temperatures and pressures, increase in temperature has a stronger effect on change in the dynamic properties of the protein globule than the increase in pressure. The conformational changes of both the deep- and shallow-sea protein models under increasing temperature and pressure are non-uniform. Our current results indicate that amino acid substitutions between shallow- and deep-water proteins only slightly affect overall stability of two proteins. Rather, they may affect the interactions of the Nip7 protein with its protein or RNA partners. PMID:25315147

  5. Communicating Science through Exhibitions

    NASA Astrophysics Data System (ADS)

    Dusenbery, P.; Harold, J.; Morrow, C.

    It is critically important for the public to better understand the scientific process. Museum exhibitions are an important part of informal science education that can effectively reach public audiences as well as school groups. They provide an important gateway for the public to learn about compelling scientific endeavors. There are many ways for scientists to help develop science exhibitions. The Space Science Institute (SSI) is a national leader in producing traveling science exhibitions and their associated educational programming (i.e. interactive websites, educator workshops, public talks, instructional materials). Two of its exhibitions, Space Weather Center and MarsQuest, are currently on tour. Another exhibition, Alien Earths, is in development. The Space Weather Center was developed in partnership with various research missions at NASA's Goddard Space Flight Center. MarsQuest is a 5000 square-foot traveling exhibition. The exhibit's second 3-year tour began this January at the Detroit Science Center. It is enabling millions of Americans to share in the excitement of the scientific exploration of Mars and to learn more about their own planet in the process. The 3,000 square-foot traveling exhibition, called Alien Earths, will bring origins-related research and discoveries to students and the American public. Alien Earths has four interrelated exhibit areas: Our Place in Space, Star Birth, PlanetQuest, and Search for Life. Exhibit visitors will explore the awesome events surrounding the birth of stars and planets; they will join scientists in the hunt for planets outside our solar system including those that may be in ``habitable zones'' around other stars; and finally they will be able to learn about how scientists are looking for signs of life beyond Earth. Besides the exhibits, SSI is also developing interactive web sites based on exhibit themes. New technologies are transforming the Web from a static medium to an interactive environment with tremendous

  6. New Hurricane Exhibit

    NASA Technical Reports Server (NTRS)

    2007-01-01

    A new exhibit in StenniSphere depicting NASA's role in hurricane prediction and research and SSC's role in helping the region recover from Hurricane Katrina. The cyclone-shaped exhibit focuses on the effects of the Aug. 29, 2005 storm and outlines how NASA is working to improve weather forecasting. Through photos, 3-D models and digital animations, the exhibit tells the story of what happened inside the storm and how NASA's scientific research can increase the accuracy of hurricane tracking and modeling.

  7. Test Control Center exhibit

    NASA Technical Reports Server (NTRS)

    2000-01-01

    Have you ever wondered how the engineers at John C. Stennis Space Center in Hancock County, Miss., test fire a Space Shuttle Main Engine? The Test Control Center exhibit at StenniSphere can answer your questions by simulating the test firing of a Space Shuttle Main Engine. A recreation of one of NASA's test control centers, the exhibit explains and portrays the 'shake, rattle and roar' that happens during a real test firing.

  8. Communicating Science through Exhibitions

    NASA Astrophysics Data System (ADS)

    Dusenbery, Paul

    2005-04-01

    It is critically important for the public to better understand the scientific process. Museum exhibitions are an important part of informal science education that can effectively reach public audiences as well as school groups. They provide an important gateway for the public to learn about compelling scientific endeavors. Science exhibitions also provide a marvelous opportunity for scientists to become engaged in the exhibit development process. The Space Science Institute (SSI) is a national leader in producing traveling science exhibitions and their associated educational programming (i.e. interactive websites, educator workshops, public talks, instructional materials). The focus of this presentation will be on two of its exhibit projects: MarsQuest (on tour for four years) and Alien Earths (its tour began early in 2005). MarsQuest is enabling millions of Americans to share in the excitement of the scientific exploration of Mars and to learn more about their own planet in the process. Alien Earths will bring origins-related research and discoveries to students and the American public. It has four interrelated exhibit areas: Our Place in Space, Star Birth, Planet Quest, and Search for Life. Exhibit visitors will explore the awesome events surrounding the birth of stars and planets; they will join scientists in the hunt for planets outside our solar system including those that may be in ``habitable zones'' around other stars; and finally they will be able to learn about how scientists are looking for signs of life beyond Earth. SSI is also developing interactive web sites based on exhibit themes. New technologies are transforming the Web from a static medium to an interactive environment with tremendous potential for informal education and inquiry-based investigations. This talk will focus on the role informal science projects play in effectively communicating science to a broad, public audience.

  9. An Extended Network of Genomic Maintenance in the Archaeon Pyrococcus abyssi Highlights Unexpected Associations between Eucaryotic Homologs

    PubMed Central

    Pluchon, Pierre-François; Fouqueau, Thomas; Crezé, Christophe; Laurent, Sébastien; Briffotaux, Julien; Hogrel, Gaëlle; Palud, Adeline; Henneke, Ghislaine; Godfroy, Anne; Hausner, Winfried; Thomm, Michael; Nicolas, Jacques; Flament, Didier

    2013-01-01

    In Archaea, the proteins involved in the genetic information processing pathways, including DNA replication, transcription, and translation, share strong similarities with those of eukaryotes. Characterizations of components of the eukaryotic-type replication machinery complex provided many interesting insights into DNA replication in both domains. In contrast, DNA repair processes of hyperthermophilic archaea are less well understood and very little is known about the intertwining between DNA synthesis, repair and recombination pathways. The development of genetic system in hyperthermophilic archaea is still at a modest stage hampering the use of complementary approaches of reverse genetics and biochemistry to elucidate the function of new candidate DNA repair gene. To gain insights into genomic maintenance processes in hyperthermophilic archaea, a protein-interaction network centred on informational processes of Pyrococcus abyssi was generated by affinity purification coupled with mass spectrometry. The network consists of 132 interactions linking 87 proteins. These interactions give insights into the connections of DNA replication with recombination and repair, leading to the discovery of new archaeal components and of associations between eucaryotic homologs. Although this approach did not allow us to clearly delineate new DNA pathways, it provided numerous clues towards the function of new molecular complexes with the potential to better understand genomic maintenance processes in hyperthermophilic archaea. Among others, we found new potential partners of the replication clamp and demonstrated that the single strand DNA binding protein, Replication Protein A, enhances the transcription rate, in vitro, of RNA polymerase. This interaction map provides a valuable tool to explore new aspects of genome integrity in Archaea and also potentially in Eucaryotes. PMID:24244547

  10. Crystal structure of product-bound complex of UDP-N-acetyl-D-mannosamine dehydrogenase from Pyrococcus horikoshii OT3

    SciTech Connect

    Pampa, K.J.; Lokanath, N.K.; Girish, T.U.; Kunishima, N.; Rai, V.R.

    2014-10-24

    Highlights: • Determined the structure of UDP-D-ManNAcADH to a resolution of 1.55 Å. • First complex structure of PhUDP-D-ManNAcADH with UDP-D-ManMAcA. • The monomeric structure consists of three distinct domains. • Cys258 acting as catalytic nucleophilic and Lys204 acts as acid/base catalyst. • Oligomeric state plays an important role for the catalytic function. - Abstract: UDP-N-acetyl-D-mannosamine dehydrogenase (UDP-D-ManNAcDH) belongs to UDP-glucose/GDP-mannose dehydrogenase family and catalyzes Uridine-diphospho-N-acetyl-D-mannosamine (UDP-D-ManNAc) to Uridine-diphospho-N-acetyl-D-mannosaminuronic acid (UDP-D-ManNAcA) through twofold oxidation of NAD{sup +}. In order to reveal the structural features of the Pyrococcus horikoshii UDP-D-ManNAcADH, we have determined the crystal structure of the product-bound enzyme by X-ray diffraction to resolution of 1.55 Å. The protomer folds into three distinct domains; nucleotide binding domain (NBD), substrate binding domain (SBD) and oligomerization domain (OD, involved in the dimerization). The clear electron density of the UDP-D-ManNAcA is observed and the residues binding are identified for the first time. Crystal structures reveal a tight dimeric polymer chains with product-bound in all the structures. The catalytic residues Cys258 and Lys204 are conserved. The Cys258 acts as catalytic nucleophile and Lys204 as acid/base catalyst. The product is directly interacts with residues Arg211, Thr249, Arg244, Gly255, Arg289, Lys319 and Arg398. In addition, the structural parameters responsible for thermostability and oligomerization of the three dimensional structure are analyzed.

  11. ADP-dependent 6-Phosphofructokinase from Pyrococcus horikoshii OT3: STRUCTURE DETERMINATION AND BIOCHEMICAL CHARACTERIZATION OF PH1645

    SciTech Connect

    Currie, Mark A.; Merino, Felipe; Skarina, Tatiana; Wong, Andrew H.Y.; Singer, Alexander; Brown, Greg; Savchenko, Alexei; Caniuguir, Andrés; Guixé, Victoria; Yakunin, Alexander F.; Jia, Zongchao

    2009-12-01

    Some hyperthermophilic archaea use a modified glycolytic pathway that employs an ADP-dependent glucokinase (ADP-GK) and an ADP-dependent phosphofructokinase (ADP-PFK) or, in the case of Methanococcus jannaschii, a bifunctional ADP-dependent glucophosphofructokinase (ADP-GK/PFK). The crystal structures of three ADP-GKs have been determined. However, there is no structural information available for ADP-PFKs or the ADP-GK/PFK. Here, we present the first crystal structure of an ADP-PFK from Pyrococcus horikoshii OT3 (PhPFK) in both apo- and AMP-bound forms determined to 2.0-{angstrom} and 1.9-{angstrom} resolution, respectively, along with biochemical characterization of the enzyme. The overall structure of PhPFK maintains a similar large and small {alpha}/{beta} domain structure seen in the ADP-GK structures. A large conformational change accompanies binding of phosphoryl donor, acceptor, or both, in all members of the ribokinase superfamily characterized thus far, which is believed to be critical to enzyme function. Surprisingly, no such conformational change was observed in the AMP-bound PhPFK structure compared with the apo structure. Through comprehensive site-directed mutagenesis of the substrate binding pocket we identified residues that were critical for both substrate recognition and the phosphotransfer reaction. The catalytic residues and many of the substrate binding residues are conserved between PhPFK and ADP-GKs; however, four key residues differ in the sugar-binding pocket, which we have shown determine the sugar-binding specificity. Using these results we were able to engineer a mutant PhPFK that mimics the ADP-GK/PFK and is able to phosphorylate both fructose 6-phosphate and glucose.

  12. Swamp to Space exhibit

    NASA Technical Reports Server (NTRS)

    2000-01-01

    The menacing-looking alligator is really harmless. It is one of the realistic props to help convince visitors that the feel of the swamp is real in StenniSphere's Swamp to Space exhibit at John C. Stennis Space Center in Hancock County, Miss. The historical section of the Swamp to Space exhibit tells the story of why and how Stennis Space Center came to be. It also pays tribute to the families who moved their homes to make way for the space age in Mississippi.

  13. Exhibition in Sight

    ERIC Educational Resources Information Center

    Wasserman, Burton

    1978-01-01

    Ludwig Mies van der Rohe is known primarily as an architect. However, he also designed chairs and tables. Discusses an exhibit held in New York City a few months ago which showed how well the famous architect achieved his goals in the area of furniture design. (Author/RK)

  14. Exhibition in Sight

    ERIC Educational Resources Information Center

    Wasserman, Burton

    1978-01-01

    One of the most offbeat exhibitions presented in the last several years was the widely celebrated Warhol-Wyeth duo show, "Portraits of Each Other", held at the Brandywine River Museum in Chadds Ford, Pennsylvania. Discusses their paintings and their diametrically different personalities. (Author/RK)

  15. Experimental silicification of the extremophilic Archaea Pyrococcus abyssi and Methanocaldococcus jannaschii: applications in the search for evidence of life in early Earth and extraterrestrial rocks.

    PubMed

    Orange, F; Westall, F; Disnar, J-R; Prieur, D; Bienvenu, N; Le Romancer, M; Défarge, Ch

    2009-09-01

    Hydrothermal activity was common on the early Earth and associated micro-organisms would most likely have included thermophilic to hyperthermophilic species. 3.5-3.3 billion-year-old, hydrothermally influenced rocks contain silicified microbial mats and colonies that must have been bathed in warm to hot hydrothermal emanations. Could they represent thermophilic or hyperthermophilic micro-organisms and if so, how were they preserved? We present the results of an experiment to silicify anaerobic, hyperthermophilic micro-organisms from the Archaea Domain Pyrococcus abyssi and Methanocaldococcus jannaschii, that could have lived on the early Earth. The micro-organisms were placed in a silica-saturated medium for periods up to 1 year. Pyrococcus abyssi cells were fossilized but the M. jannaschii cells lysed naturally after the exponential growth phase, apart from a few cells and cell remains, and were not silicified although their extracellular polymeric substances were. In this first simulated fossilization of archaeal strains, our results suggest that differences between species have a strong influence on the potential for different micro-organisms to be preserved by fossilization and that those found in the fossil record represent probably only a part of the original diversity. Our results have important consequences for biosignatures in hydrothermal or hydrothermally influenced deposits on Earth, as well as on early Mars, as environmental conditions were similar on the young terrestrial planets and traces of early Martian life may have been similarly preserved as silicified microfossils.

  16. Experimental silicification of the extremophilic Archaea Pyrococcus abyssi and Methanocaldococcus jannaschii: applications in the search for evidence of life in early Earth and extraterrestrial rocks.

    PubMed

    Orange, F; Westall, F; Disnar, J-R; Prieur, D; Bienvenu, N; Le Romancer, M; Défarge, Ch

    2009-09-01

    Hydrothermal activity was common on the early Earth and associated micro-organisms would most likely have included thermophilic to hyperthermophilic species. 3.5-3.3 billion-year-old, hydrothermally influenced rocks contain silicified microbial mats and colonies that must have been bathed in warm to hot hydrothermal emanations. Could they represent thermophilic or hyperthermophilic micro-organisms and if so, how were they preserved? We present the results of an experiment to silicify anaerobic, hyperthermophilic micro-organisms from the Archaea Domain Pyrococcus abyssi and Methanocaldococcus jannaschii, that could have lived on the early Earth. The micro-organisms were placed in a silica-saturated medium for periods up to 1 year. Pyrococcus abyssi cells were fossilized but the M. jannaschii cells lysed naturally after the exponential growth phase, apart from a few cells and cell remains, and were not silicified although their extracellular polymeric substances were. In this first simulated fossilization of archaeal strains, our results suggest that differences between species have a strong influence on the potential for different micro-organisms to be preserved by fossilization and that those found in the fossil record represent probably only a part of the original diversity. Our results have important consequences for biosignatures in hydrothermal or hydrothermally influenced deposits on Earth, as well as on early Mars, as environmental conditions were similar on the young terrestrial planets and traces of early Martian life may have been similarly preserved as silicified microfossils. PMID:19656214

  17. Starship 2040 Exhibit

    NASA Technical Reports Server (NTRS)

    2002-01-01

    This photograph shows onlookers viewing displays within the Starship 2040 exhibit on display at Joe Davis Stadium in Huntsville, Alabama. Developed by the Space Transportation Directorate at Marshall Space Flight Center (MSFC), the Starship 2040 exhibit is housed in a 48-ft (14.6-m) tractor and trailer rig, permitting it to travel around the Nation, demonstrating NASA's vision of what commercial spaceflight might be like 40 years from now. All the irnovations suggested aboard the exhibit (automated vehicle health monitoring systems, high-energy propulsion drive, navigational aids, and emergency and safety systems) are based on concepts and technologies now being studied at NASA Centers and partner institutions around the Nation. NASA is the Nation's premier agency for development of the space transportation system, including future-generation reusable launch vehicles. Such systems, the keys to a 'real' Starship 2040, require revolutionary advances in critical aerospace technologies, from thermal, magnetic, chemical, and propellantless propulsion systems to new energy sources such as space solar power or antimatter propulsion. These and other advances are now being studied, developed, and tested at NASA field centers and partner institutions all over the Nation.

  18. Starship 2040 Exhibit

    NASA Technical Reports Server (NTRS)

    2002-01-01

    This photograph shows Justin Varnadore, son of a Marshall TV employee, at the controls of one of the many displays within the Starship 2040 exhibit on display at Joe Davis Stadium in Huntsville, Alabama. Developed by the Space Transportation Directorate at Marshall Space Flight Center (MSFC), the Starship 2040 exhibit is housed in a 48-ft (14.6-m) tractor and trailer rig, permitting it to travel around the Nation, demonstrating NASA's vision of what commercial spaceflight might be like 40 years from now. All the irnovations suggested aboard the exhibit (automated vehicle health monitoring systems, high-energy propulsion drive, navigational aids, and emergency and safety systems) are based on concepts and technologies now being studied at NASA Centers and partner institutions around the Nation. NASA is the Nation's premier agency for development of the space transportation system, including future-generation reusable launch vehicles. Such systems, the keys to a 'real' Starship 2040, require revolutionary advances in critical aerospace technologies, from thermal, magnetic, chemical, and propellantless propulsion systems to new energy sources such as space solar power or antimatter propulsion. These and other advances are now being studied, developed, and tested at NASA field centers and partner institutions all over the Nation.

  19. Starship 2040 Exhibit

    NASA Technical Reports Server (NTRS)

    2001-01-01

    This photograph shows the Starship 2040 leaving the Marshall Space Flight Center (MSFC) for the exhibit site. Developed by the Space Transportation Directorate at MSFC, the Starship 2040 exhibit is housed in a 48-ft (14.6-m) tractor and trailer rig, permitting it to travel around the Nation, demonstrating NASA's vision of what commercial spaceflight might be like 40 years from now. All the irnovations suggested aboard the exhibit, automated vehicle health monitoring systems, high-energy propulsion drive, navigational aids and emergency and safety systems, are based on concepts and technologies now being studied at NASA Centers and partner institutions around the Nation. NASA is the nation's premier agency for development of the space transportation system, including future-generation reusable launch vehicles. Such systems, the keys to a 'real' Starship 2040, require revolutionary advances in critical aerospace technologies, from thermal, magnetic, chemical, and propellantless propulsion systems to new energy sources such as space solar power or antimatter propulsion. These and other advances are now being studied, developed, and tested at NASA field centers and partner institutions all over the Nation.

  20. Online Exhibits & Concept Maps

    NASA Astrophysics Data System (ADS)

    Douma, M.

    2009-12-01

    Presenting the complexity of geosciences to the public via the Internet poses a number of challenges. For example, utilizing various - and sometimes redundant - Web 2.0 tools can quickly devour limited time. Do you tweet? Do you write press releases? Do you create an exhibit or concept map? The presentation will provide participants with a context for utilizing Web 2.0 tools by briefly highlighting methods of online scientific communication across several dimensions. It will address issues of: * breadth and depth (e.g. from narrow topics to well-rounded views), * presentation methods (e.g. from text to multimedia, from momentary to enduring), * sources and audiences (e.g. for experts or for the public, content developed by producers to that developed by users), * content display (e.g. from linear to non-linear, from instructive to entertaining), * barriers to entry (e.g. from an incumbent advantage to neophyte accessible, from amateur to professional), * cost and reach (e.g. from cheap to expensive), and * impact (e.g. the amount learned, from anonymity to brand awareness). Against this backdrop, the presentation will provide an overview of two methods of online information dissemination, exhibits and concept maps, using the WebExhibits online museum (www.webexhibits.org) and SpicyNodes information visualization tool (www.spicynodes.org) as examples, with tips on how geoscientists can use either to communicate their science. Richly interactive online exhibits can serve to engage a large audience, appeal to visitors with multiple learning styles, prompt exploration and discovery, and present a topic’s breadth and depth. WebExhibits, which was among the first online museums, delivers interactive information, virtual experiments, and hands-on activities to the public. While large, multidisciplinary exhibits on topics like “Color Vision and Art” or “Calendars Through the Ages” require teams of scholars, user interface experts, professional writers and editors

  1. Space Shuttle Cockpit exhibit

    NASA Technical Reports Server (NTRS)

    2000-01-01

    Want to sit in the cockpit of the Space Shuttle and watch astronauts work in outer space? At StenniSphere, you can do that and much more. StenniSphere, the visitor center at John C. Stennis Space Center in Hancock County, Miss., presents 14,000-square-feet of interactive exhibits that depict America's race for space as well as a glimpse of the future. StenniSphere is open free of charge from 9 a.m. to 5 p.m. daily.

  2. Smithsonian climate change exhibits

    NASA Astrophysics Data System (ADS)

    Kumar, Mohi

    2006-05-01

    Two new museum exhibits, ``Arctic: A Friend Acting Strangely'' and ``Atmosphere: Change is in the Air'' opened 15 April at the Smithsonian Institution's National Museum of Natural History in Washington, D.C., in partnership with the U.S. National Oceanic and Atmospheric Administration, NASA, and the U.S. National Science Foundation. In ``Arctic: A Friend Acting Strangely,'' anecdotes from indigenous polar people reveal how climate changes have affected life within the last 50 years. For example, as permafrost melts and sea ice shrinks, plant distributions and animal migration patterns are changing, severely affecting culture.

  3. Starship 2040 Exhibit

    NASA Technical Reports Server (NTRS)

    2002-01-01

    This photograph shows the Starship 2040 on display at Joe Davis Stadium in Huntsville, Alabama. Developed by the Space Transportation Directorate at Marshall Space Flight Center (MSFC), the Starship 2040 exhibit is housed in a 48-ft (14.6-m) tractor and trailer rig, permitting it to travel around the Nation, demonstrating NASA's vision of what commercial spaceflight might be like 40 years from now. All the irnovations suggested aboard the exhibit (automated vehicle health monitoring systems, high-energy propulsion drive, navigational aids, and emergency and safety systems) are based on concepts and technologies now being studied at NASA Centers and partner institutions around the Nation. NASA is the Nation's premier agency for development of the space transportation system, including future-generation reusable launch vehicles. Such systems, the keys to a 'real' Starship 2040, require revolutionary advances in critical aerospace technologies, from thermal, magnetic, chemical, and propellantless propulsion systems to new energy sources such as space solar power or antimatter propulsion. These and other advances are now being studied, developed, and tested at NASA field centers and partner institutions all over the Nation.

  4. Dimeric 3-phosphoglycerate kinases from hyperthermophilic Archaea. Cloning, sequencing and expression of the 3-phosphoglycerate kinase gene of Pyrococcus woesei in Escherichia coli and characterization of the protein. Structural and functional comparison with the 3-phosphoglycerate kinase of Methanothermus fervidus.

    PubMed

    Hess, D; Krüger, K; Knappik, A; Palm, P; Hensel, R

    1995-10-01

    The gene coding for the 3-phosphoglycerate kinase (EC 2.7.2.3) of Pyrococcus woesei was cloned and sequenced. The gene sequence comprises 1230 bp coding for a polypeptide with the theoretical M(r) of 46,195. The deduced protein sequence exhibits a high similarity (46.1% and 46.6% identity) to the other known archaeal 3-phosphoglycerate kinases of Methanobacterium bryantii and Methanothermus fervidus [Fabry, S., Heppner, P., Dietmaier, W. & Hensel, R. (1990) Gene 91, 19-25]. By comparing the 3-phosphoglycerate kinase sequences of the mesophilic and the two thermophilic Archaea, trends in thermoadaptation were confirmed that could be deduced from comparisons of glyceraldehyde-3-phosphate dehydrogenase sequences from the same organisms [Zwickl, P., Fabry, S., Bogedain, C., Haas, A. & Hensel, R. (1990) J. Bacteriol. 172, 4329-4338]. With increasing temperature the average hydrophobicity and the portion of aromatic residues increases, whereas the chain flexibility as well as the content in chemically labile residues (Asn, Cys) decreases. To study the phenotypic properties of the 3-phosphoglycerate kinases from thermophilic Archaea in more detail, the 3-phosphoglycerate kinase genes from P. woesei and M. fervidus were expressed in Escherichia coli. Comparisons of kinetic and molecular properties of the enzymes from the original organisms and from E. coli indicate that the proteins expressed in the mesophilic host are folded correctly. Besides their higher thermostability according to their origin from hyperthermophilic organisms, both enzymes differ from their bacterial and eucaryotic homologues mainly in two respects. (a) The 3-phosphoglycerate kinases from P. woesei and M. fervidus are homomeric dimers in their native state contrary to all other known 3-phosphoglycerate kinases, which are monomers including the enzyme from the mesophilic Archaeum M. bryantii. (b) Monovalent cations are essential for the activity of both archaeal enzymes with K+ being significantly more

  5. Dimeric 3-phosphoglycerate kinases from hyperthermophilic Archaea. Cloning, sequencing and expression of the 3-phosphoglycerate kinase gene of Pyrococcus woesei in Escherichia coli and characterization of the protein. Structural and functional comparison with the 3-phosphoglycerate kinase of Methanothermus fervidus.

    PubMed

    Hess, D; Krüger, K; Knappik, A; Palm, P; Hensel, R

    1995-10-01

    The gene coding for the 3-phosphoglycerate kinase (EC 2.7.2.3) of Pyrococcus woesei was cloned and sequenced. The gene sequence comprises 1230 bp coding for a polypeptide with the theoretical M(r) of 46,195. The deduced protein sequence exhibits a high similarity (46.1% and 46.6% identity) to the other known archaeal 3-phosphoglycerate kinases of Methanobacterium bryantii and Methanothermus fervidus [Fabry, S., Heppner, P., Dietmaier, W. & Hensel, R. (1990) Gene 91, 19-25]. By comparing the 3-phosphoglycerate kinase sequences of the mesophilic and the two thermophilic Archaea, trends in thermoadaptation were confirmed that could be deduced from comparisons of glyceraldehyde-3-phosphate dehydrogenase sequences from the same organisms [Zwickl, P., Fabry, S., Bogedain, C., Haas, A. & Hensel, R. (1990) J. Bacteriol. 172, 4329-4338]. With increasing temperature the average hydrophobicity and the portion of aromatic residues increases, whereas the chain flexibility as well as the content in chemically labile residues (Asn, Cys) decreases. To study the phenotypic properties of the 3-phosphoglycerate kinases from thermophilic Archaea in more detail, the 3-phosphoglycerate kinase genes from P. woesei and M. fervidus were expressed in Escherichia coli. Comparisons of kinetic and molecular properties of the enzymes from the original organisms and from E. coli indicate that the proteins expressed in the mesophilic host are folded correctly. Besides their higher thermostability according to their origin from hyperthermophilic organisms, both enzymes differ from their bacterial and eucaryotic homologues mainly in two respects. (a) The 3-phosphoglycerate kinases from P. woesei and M. fervidus are homomeric dimers in their native state contrary to all other known 3-phosphoglycerate kinases, which are monomers including the enzyme from the mesophilic Archaeum M. bryantii. (b) Monovalent cations are essential for the activity of both archaeal enzymes with K+ being significantly more

  6. Crystallization and preliminary X-ray crystallographic analysis of a conserved domain in plants and prokaryotes from Pyrococcus horikoshii OT3

    SciTech Connect

    Lin, Linyen; Nakano, Hiroaki; Uchiyama, Susumu; Fujimoto, Satoru; Matsunaga, Sachihiro; Nakamura, Shota; Kobayashi, Yuji; Ohkubo, Tadayasu; Fukui, Kiichi

    2005-04-01

    A plant- and prokaryote-conserved domain (PPC) has been crystallized. The crystal diffracted to 1.7 Å resolution and belonged to space group P6{sub 3}22. A plant- and prokaryote-conserved domain (PPC) has previously been found in AT-hook motif nuclear localized protein 1 (AHL1) localized in the nuclear matrix of Arabidopsis thaliana (AtAHL1). AtAHL1 has a DNA-binding function. Mutation analyses of AtAHL1 has previously revealed that the hydrophobic region of the PPC domain is essential for its nuclear localization. In this study, the PPC of the hyperthermophilic archaebacterium Pyrococcus horikoshii (PhPPC) was crystallized using the hanging-drop vapour-diffusion method. The crystals belonged to the hexagonal space group P6{sub 3}22, with unit-cell parameters a = b = 53.69, c = 159.2 Å. Data were obtained at 100 K, with diffraction being observed to a resolution of 1.7 Å. A complete data set from crystals of the SeMet-substituted protein was also obtained.

  7. The TET2 and TET3 aminopeptidases from Pyrococcus horikoshii form a hetero-subunit peptidasome with enhanced peptide destruction properties.

    PubMed

    Appolaire, Alexandre; Durá, M Asunción; Ferruit, Mylène; Andrieu, Jean-Pierre; Godfroy, Anne; Gribaldo, Simonetta; Franzetti, Bruno

    2014-11-01

    TET aminopeptidases assemble as large homo-dodecameric complexes. The reason why prokaryotic genomes often encode a diverse set of TET peptidases homologues remains unclear. In the archaeon Pyrococcus horikoshii, PhTET1, PhTET2 and PhTET3 homo-oligomeric particles have been proposed to work in concert to breakdown intracellular polypeptides. When coexpressed in Escherichia coli, the PhTET2 and PhTET3 proteins were found to assemble efficiently as heteromeric complexes. Biophysical analysis demonstrated that these particles possess the same quaternary structure as the homomeric TET dodecamers. The same hetero-oligomeric complexes were immunodetected in P. horikoshii cell extracts analysed by sucrose gradient fractionation and ion exchange chromatography. The biochemical activity of a purified hetero-oligomeric TET particle, assessed on chromogenic substrates and on a complex mixture of peptides, reveals that it displays higher efficiency than an equivalent combination of homo-oligomeric TET particles. Interestingly, phylogenetic analysis shows that PhTET2 and PhTET3 are paralogous proteins that arose from gene duplication in the ancestor of Thermococcales. Together, these results establish that the PhTET2 and PhTET3 proteins are two subunits of the same enzymatic complex aimed at the destruction of polypeptidic chains of very different composition. This is the first report for such a mechanism intended to improve multi-enzymatic complex efficiency among exopeptidases.

  8. Isolation and characterization of a heat-stable pullulanase from the hyperthermophilic archaeon Pyrococcus woesei after cloning and expression of its gene in Escherichia coli.

    PubMed Central

    Rüdiger, A; Jorgensen, P L; Antranikian, G

    1995-01-01

    The gene encoding an extremely heat-stable pullulanase from the hyperthermophilic archaeon Pyrococcus woesei was cloned and expressed in Escherichia coli. Purification of the enzyme to homogeneity was achieved after heat treatment of the recombinant E. coli cells, affinity chromatography on a maltotriose-coupled Sepharose 6B column, and anion-exchange chromatography on Mono Q. The pullulanase, which was purified 90-fold with a final yield of 15%, is composed of a single polypeptide chain with a molecular mass of 90 kDa. The enzyme is optimally active at 100 degrees C and pH 6.0 and shows 40% activity at 120 degrees C. Enzyme activation up to 370% is achieved in the presence of calcium ions and reducing agents such as beta-mercaptoethanol and dithiothreitol, whereas N-bromosuccinimide and alpha-cyclodextrin are inhibitory. The high rigidity of the heat-stable enzyme is demonstrated by fluorescence spectroscopic studies in the presence of denaturing agents such as sodium dodecyl sulfate. At temperatures above 80 degrees C, the enzyme seems to switch from the compact to the unfolded form, which is accompanied by an apparent shift in the molecular mass from 45 to 90 kDa. PMID:7574598

  9. Exhibitions: Facing Outward, Pointing Inward

    ERIC Educational Resources Information Center

    McDonald, Joseph P.

    2007-01-01

    The Coalition of Essential Schools (CES) Exhibitions Project of the early 1990s produced a range of work that continues to inform the practice of using exhibitions as a "360 degree" method of transforming teaching and learning, community connections, school design, and assessment. Among that work was this paper coupling the origins of exhibitions…

  10. Against the Odds Exhibition Opens

    MedlinePlus

    ... LA and Vox Populi organizations. Photo courtesy of Bill Branson At the exhibition, HIV and AIDS were topics addressed by Dr. Victoria Cargill (right), Director of Clinical Studies and Director of Minority ...

  11. Greenhouse Earth: A Traveling Exhibition

    SciTech Connect

    Booth, W.H.; Caesar, S.

    1992-09-01

    The Franklin Institute Science Museum provided an exhibit entitled the Greenhouse Earth: A Traveling Exhibition. This 3500 square-foot exhibit on global climate change was developed in collaboration with the Association of Science-Technology Centers. The exhibit opened at The Franklin Institute on February 14, 1992, welcoming 291,000 visitors over its three-month stay. During its three-year tour, Greenhouse Earth will travel to ten US cities, reaching two million visitors. Greenhouse Earth aims to deepen public understanding of the scientific issues of global warming and the conservation measures that can be taken to slow its effects. The exhibit features hands-on exhibitry, interactive computer programs and videos, a theater production, a demonstration cart,'' guided tours, and lectures. supplemental educational programs at the Institute included a teachers preview, a symposium on climate change, and a satellite field trip.'' The development of Greenhouse Earth included front-end and formative evaluation procedures. Evaluation includes interviews with visitors, prototypes, and summative surveys for participating museums. During its stay in Philadelphia, Greenhouse Earth was covered by the local and national press, with reviews in print and broadcast media. Greenhouse Earth is the first large-scale museum exhibit to address global climate change.

  12. Considering High-Tech Exhibits?

    ERIC Educational Resources Information Center

    Routman, Emily

    1994-01-01

    Discusses a variety of high-tech exhibit media used in The Living World, an educational facility operated by The Saint Louis Zoo. Considers the strengths and weaknesses of holograms, video, animatronics, video-equipped microscopes, and computer interactives. Computer interactives are treated with special attention. (LZ)

  13. Survival and growth of two heterotrophic hydrothermal vent archaea, Pyrococcus strain GB-D and Thermococcus fumicolans, under low pH and high sulfide concentrations in combination with high temperature and pressure regimes.

    PubMed

    Edgcomb, Virginia P; Molyneaux, Stephen J; Böer, Simone; Wirsen, Carl O; Saito, Mak; Atkins, Michael S; Lloyd, Karen; Teske, Andreas

    2007-03-01

    Growth and survival of hyperthermophilic archaea in their extreme hydrothermal vent and subsurface environments are controlled by chemical and physical key parameters. This study examined the effects of elevated sulfide concentrations, temperature, and acidic pH on growth and survival of two hydrothermal vent archaea (Pyrococcus strain GB-D and Thermococcus fumicolans) under high temperature and pressure regimes. These two strains are members of the Thermococcales, a family of hyperthermophilic, heterotrophic, sulfur-reducing archaea that occur in high densities at vent sites. As actively growing cells, these two strains tolerated regimes of pH, pressure, and temperature that were in most cases not tolerated under severe substrate limitation. A moderate pH of 5.5-7 extends their survival and growth range over a wider range of sulfide concentrations, temperature and pressure, relative to lower pH conditions. T. fumicolans and Pyrococcus strain GB-D grew under very high pressures that exceeded in-situ pressures typical of hydrothermal vent depths, and included deep subsurface pressures. However, under the same conditions, but in the absence of carbon substrates and electron acceptors, survival was generally lower, and decreased rapidly when low pH stress was combined with high pressure and high temperature.

  14. Balamuthia mandrillaris exhibits metalloprotease activities.

    PubMed

    Matin, Abdul; Stins, Monique; Kim, Kwang Sik; Khan, Naveed Ahmed

    2006-06-01

    Balamuthia mandrillaris is a recently identified protozoan pathogen that can cause fatal granulomatous encephalitis. However, the pathogenesis and pathophysiology of B. mandrillaris encephalitis remain unclear. Because proteases may play a role in the central nervous system (CNS) pathology, we used spectrophotometric, cytopathic and zymographic assays to assess protease activities of B. mandrillaris. Using two clinical isolates of B. mandrillaris (from human and baboon), we observed that B. mandrillaris exhibits protease activities. Zymographic assays revealed major protease bands of approximate molecular weights in the region of 40-50 kDa on sodium dodecyl sulfate-polyacrylamide gels using gelatin as substrate. The protease bands were inhibited with 1,10-phenanthroline, suggesting metallo-type proteases. The proteolytic activities were observed over a pH range of 5-11 with maximum activity at neutral pH and at 42 degrees C. Balamuthia mandrillaris proteases exhibit properties to degrade extracellular matrix (ECM), which provide structural and functional support to the brain tissue. This is shown by degradation of collagen I and III (major components of collagenous ECM), elastin (elastic fibrils of ECM), plasminogen (involved in proteolytic degradation of ECM), as well as other substrates such as casein and gelatin but not haemoglobin. However, these proteases exhibited a minimal role in B. mandrillaris-mediated host cell death in vitro using human brain microvascular endothelial cells (HBMECs). This was shown using broad-spectrum matrix metalloprotease inhibitors, GM 6001 and GM 1489, which had no effect on B. mandrillaris-mediated HBMEC cytotoxicity. This is the first demonstration that B. mandrillaris exhibits metalloproteases, which may play important role(s) in the ECM degradation and thus in CNS pathology. PMID:16706791

  15. Crows spontaneously exhibit analogical reasoning.

    PubMed

    Smirnova, Anna; Zorina, Zoya; Obozova, Tanya; Wasserman, Edward

    2015-01-19

    Analogical reasoning is vital to advanced cognition and behavioral adaptation. Many theorists deem analogical thinking to be uniquely human and to be foundational to categorization, creative problem solving, and scientific discovery. Comparative psychologists have long been interested in the species generality of analogical reasoning, but they initially found it difficult to obtain empirical support for such thinking in nonhuman animals (for pioneering efforts, see [2, 3]). Researchers have since mustered considerable evidence and argument that relational matching-to-sample (RMTS) effectively captures the essence of analogy, in which the relevant logical arguments are presented visually. In RMTS, choice of test pair BB would be correct if the sample pair were AA, whereas choice of test pair EF would be correct if the sample pair were CD. Critically, no items in the correct test pair physically match items in the sample pair, thus demanding that only relational sameness or differentness is available to support accurate choice responding. Initial evidence suggested that only humans and apes can successfully learn RMTS with pairs of sample and test items; however, monkeys have subsequently done so. Here, we report that crows too exhibit relational matching behavior. Even more importantly, crows spontaneously display relational responding without ever having been trained on RMTS; they had only been trained on identity matching-to-sample (IMTS). Such robust and uninstructed relational matching behavior represents the most convincing evidence yet of analogical reasoning in a nonprimate species, as apes alone have spontaneously exhibited RMTS behavior after only IMTS training.

  16. Collaborative virtual environments art exhibition

    NASA Astrophysics Data System (ADS)

    Dolinsky, Margaret; Anstey, Josephine; Pape, Dave E.; Aguilera, Julieta C.; Kostis, Helen-Nicole; Tsoupikova, Daria

    2005-03-01

    This panel presentation will exhibit artwork developed in CAVEs and discuss how art methodologies enhance the science of VR through collaboration, interaction and aesthetics. Artists and scientists work alongside one another to expand scientific research and artistic expression and are motivated by exhibiting collaborative virtual environments. Looking towards the arts, such as painting and sculpture, computer graphics captures a visual tradition. Virtual reality expands this tradition to not only what we face, but to what surrounds us and even what responds to our body and its gestures. Art making that once was isolated to the static frame and an optimal point of view is now out and about, in fully immersive mode within CAVEs. Art knowledge is a guide to how the aesthetics of 2D and 3D worlds affect, transform, and influence the social, intellectual and physical condition of the human body through attention to psychology, spiritual thinking, education, and cognition. The psychological interacts with the physical in the virtual in such a way that each facilitates, enhances and extends the other, culminating in a "go together" world. Attention to sharing art experience across high-speed networks introduces a dimension of liveliness and aliveness when we "become virtual" in real time with others.

  17. Crows spontaneously exhibit analogical reasoning.

    PubMed

    Smirnova, Anna; Zorina, Zoya; Obozova, Tanya; Wasserman, Edward

    2015-01-19

    Analogical reasoning is vital to advanced cognition and behavioral adaptation. Many theorists deem analogical thinking to be uniquely human and to be foundational to categorization, creative problem solving, and scientific discovery. Comparative psychologists have long been interested in the species generality of analogical reasoning, but they initially found it difficult to obtain empirical support for such thinking in nonhuman animals (for pioneering efforts, see [2, 3]). Researchers have since mustered considerable evidence and argument that relational matching-to-sample (RMTS) effectively captures the essence of analogy, in which the relevant logical arguments are presented visually. In RMTS, choice of test pair BB would be correct if the sample pair were AA, whereas choice of test pair EF would be correct if the sample pair were CD. Critically, no items in the correct test pair physically match items in the sample pair, thus demanding that only relational sameness or differentness is available to support accurate choice responding. Initial evidence suggested that only humans and apes can successfully learn RMTS with pairs of sample and test items; however, monkeys have subsequently done so. Here, we report that crows too exhibit relational matching behavior. Even more importantly, crows spontaneously display relational responding without ever having been trained on RMTS; they had only been trained on identity matching-to-sample (IMTS). Such robust and uninstructed relational matching behavior represents the most convincing evidence yet of analogical reasoning in a nonprimate species, as apes alone have spontaneously exhibited RMTS behavior after only IMTS training. PMID:25532894

  18. 17 CFR 232.102 - Exhibits.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... Regulation S-T (17 CFR 232.311(b)). (b) Amendments to all exhibits shall be filed in electronic format... designation “CE” (confirming electronic) should be placed next to the listed exhibit in the exhibit index....

  19. A complex standard for protein identification, designed by evolution.

    PubMed

    Vaudel, Marc; Burkhart, Julia M; Breiter, Daniela; Zahedi, René P; Sickmann, Albert; Martens, Lennart

    2012-10-01

    Shotgun proteomic investigations rely on the algorithmic assignment of mass spectra to peptides. The quality of these matches is therefore a cornerstone in the analysis and has been the subject of numerous recent developments. In order to establish the benefits of novel algorithms, they are applied to reference samples of known content. However, these were recently shown to be either too simple to resemble typical real-life samples or as leading to results of lower accuracy as the method itself. Here, we describe how to use the proteome of Pyrococcus furiosus , a hyperthermophile, as a standard to evaluate proteomics identification workflows. Indeed, we prove that the Pyrococcus furiosus proteome provides a valid method for detecting random hits, comparable to the decoy databases currently in popular use, but we also prove that the Pyrococcus furiosus proteome goes squarely beyond the decoy approach by also providing many hundreds of highly reliable true positive hits. Searching the Pyrococcus furiosus proteome can thus be used as a unique test that provides the ability to reliably detect both false positives as well as proteome-scale true positives, allowing the rigorous testing of identification algorithms at the peptide and protein level.

  20. Exhibits Enhanced by Stand-Alone Computers.

    ERIC Educational Resources Information Center

    Van Rennes, Eve C.

    Both the development and evaluation of one of a set of computer programs designed for use by visitors as adjuncts to museum exhibits are described. Museum displays used were (1) a static, behind-glass exhibit on evolution; (2) a hands-on primitive stone age tools exhibit; and (3) a Foucault pendulum. A computer placed next to each exhibit served…

  1. Science Education Through a Museum Exhibit

    ERIC Educational Resources Information Center

    Chaparian, Azad; And Others

    1973-01-01

    Describes the polywater exhibit at the Worcester Science Center in Massachusetts. Curiosity and interest are stimulated in young people by allowing them to handle the materials in the exhibit and by providing them with instructions for making polywater. (JR)

  2. Learning by Doing, Creating a Museum Exhibit.

    ERIC Educational Resources Information Center

    Main, Sarah; Kallquist, Dierdre

    2000-01-01

    Describes an exhibit called Kid's Kitchen, built within a major exhibit called Biodiversity: Life Supporting Life, in order to discuss environmental prompts hidden within the kitchen designed to surprise students and get them thinking. (ASK)

  3. 32 CFR 705.24 - Exhibits.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... shopping centers, malls, etc.) provided it is clearly established that such areas are places the general... exhibits may be forwarded to the Navy Recruiting Exhibit Center via the local Navy recruiter with an information copy to the Chief of Information. The primary mission of the Navy Recruiting Exhibit Center is...

  4. 32 CFR 705.24 - Exhibits.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... shopping centers, malls, etc.) provided it is clearly established that such areas are places the general... exhibits may be forwarded to the Navy Recruiting Exhibit Center via the local Navy recruiter with an information copy to the Chief of Information. The primary mission of the Navy Recruiting Exhibit Center is...

  5. 32 CFR 705.24 - Exhibits.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... shopping centers, malls, etc.) provided it is clearly established that such areas are places the general... exhibits may be forwarded to the Navy Recruiting Exhibit Center via the local Navy recruiter with an information copy to the Chief of Information. The primary mission of the Navy Recruiting Exhibit Center is...

  6. 32 CFR 705.24 - Exhibits.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... shopping centers, malls, etc.) provided it is clearly established that such areas are places the general... exhibits may be forwarded to the Navy Recruiting Exhibit Center via the local Navy recruiter with an information copy to the Chief of Information. The primary mission of the Navy Recruiting Exhibit Center is...

  7. 29 CFR 2200.70 - Exhibits.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Relating to Labor (Continued) OCCUPATIONAL SAFETY AND HEALTH REVIEW COMMISSION RULES OF PROCEDURE Hearings... separate file designated for rejected exhibits. (e) Return of physical exhibits. A party may on motion request the return of a physical exhibit within 30 days after expiration of the time for filing a...

  8. 29 CFR 2200.70 - Exhibits.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Relating to Labor (Continued) OCCUPATIONAL SAFETY AND HEALTH REVIEW COMMISSION RULES OF PROCEDURE Hearings... separate file designated for rejected exhibits. (e) Return of physical exhibits. A party may on motion request the return of a physical exhibit within 30 days after expiration of the time for filing a...

  9. 29 CFR 2200.70 - Exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Relating to Labor (Continued) OCCUPATIONAL SAFETY AND HEALTH REVIEW COMMISSION RULES OF PROCEDURE Hearings... separate file designated for rejected exhibits. (e) Return of physical exhibits. A party may on motion request the return of a physical exhibit within 30 days after expiration of the time for filing a...

  10. A new crystal form of a hyperthermophilic endocellulase

    SciTech Connect

    Kataoka, Misumi; Ishikawa, Kazuhiko

    2014-06-18

    The hyperthermostable endocellulase from P. furiosus was crystallized at pH 5.5. The new crystal form has symmetry consistent with space group C2 and exhibits a structure different from that of the protein crystallized at pH 9.0. The hyperthermophilic glycoside hydrolase family endocellulase 12 from the archaeon Pyrococcus furiosus (EGPf; Gene ID PF0854; EC 3.2.1.4) catalyzes the hydrolytic cleavage of the β-1,4-glucosidic linkage in β-glucan in lignocellulose biomass. A crystal of EGPf was previously prepared at pH 9.0 and its structure was determined at an atomic resolution of 1.07 Å. This article reports the crystallization of EGPf at the more physiologically relevant pH of 5.5. Structure determination showed that this new crystal form has the symmetry of space group C2. Two molecules of the enzyme are observed in the asymmetric unit. Crystal packing is weak at pH 5.5 owing to two flexible interfaces between symmetry-related molecules. Comparison of the EGPf structures obtained at pH 9.0 and pH 5.5 reveals a significant conformational difference at the active centre and in the surface loops. The interfaces in the vicinity of the flexible surface loops impact the quality of the EGPf crystal.

  11. Superconductive microstrip exhibiting negative differential resistivity

    DOEpatents

    Huebener, R.P.; Gallus, D.E.

    1975-10-28

    A device capable of exhibiting negative differential electrical resistivity over a range of values of current and voltage is formed by vapor- depositing a thin layer of a material capable of exhibiting superconductivity on an insulating substrate, establishing electrical connections at opposite ends of the deposited strip, and cooling the alloy into its superconducting range. The device will exhibit negative differential resistivity when biased in the current- induced resistive state.

  12. Encountering Nanotechnology in an Interactive Exhibition

    ERIC Educational Resources Information Center

    Murriello, Sandra E.; Knobel, Marcelo

    2008-01-01

    This article offers findings from a learning sciences-informed evaluation of a nanoscience and nanotechnology exhibition called Nano-Aventura (NanoAdventure), based on four interactive-collaborative games and two narrated videos. This traveling exhibition was developed in Brazil by the Museu Exploratorio de Ciencias for children and teenagers…

  13. Strategies for Determining Exhibit Effectiveness. Final Report.

    ERIC Educational Resources Information Center

    Shettel, Harris H.; And Others

    This project was designed to develop research strategies and hypotheses for evaluating the effectiveness of exhibits. An exhibit on the role of the Federal Government in science and technology was used as the subject matter. Two basic groups of viewers were used, casual viewers and paid experimental viewers. Both were tested on knowledge gained…

  14. Memory and Mourning: An Exhibit History

    ERIC Educational Resources Information Center

    Eberle, Scott G.

    2005-01-01

    Mounted by the Strong Museum in Rochester, New York, in 1993, and traveling nationally thereafter, the exhibit Memory and Mourning provided historical and contemporary perspectives to help museum guests explore their own reactions to loss and grief. In the process the exhibit's development team encountered a range of philosophical, historical,…

  15. Science Fiction Exhibits as STEM Gateways

    NASA Astrophysics Data System (ADS)

    Robie, Samantha

    Women continue to hold less than a quarter of all STEM jobs in the United States, prompting many museums to develop programs and exhibits with the express goal of interesting young girls in scientific fields. At the same time, a number of recent museum exhibits have harnessed the popularity of pop culture and science fiction in order to interest general audiences in STEM subject matter, as well as using the exhibits as springboards to expand or shift mission goals and focus. Because science fiction appears to be successful at raising interest in STEM fields, it may be an effective way to garner the interest of young girls in STEM in particular. This research seeks to describe the ways in which museums are currently using science fiction exhibits to interest young girls in STEM fields and careers. Research focused on four institutions across the country hosting three separate exhibits, and included staff interviews and content analysis of exhibit descriptions, promotional materials, a summative evaluation and supplementary exhibit productions. In some ways, science fiction exhibits do serve young girls, primarily through the inclusion of female role models, staff awareness, and prototype testing to ensure interactives are attractive to girls as well as to boys. However, STEM appears to be underutilized, which may be partly due to a concern within the field that the outcome of targeting a specific gender could be construed as "stereotyping".

  16. Learning4Life on the Exhibit Floor

    ERIC Educational Resources Information Center

    Sullivan, Margaret

    2009-01-01

    The exhibit floor is a wealth of knowledge. One can read, view, and listen to information presented in many formats. Somewhere on the exhibit floor there are experts on every topic, ready and waiting for one's questions. But like any research topic, frequently a structured search is required to find the best answers. This article discusses how to…

  17. Exhibit of School Architecture, 1997. Special Section.

    ERIC Educational Resources Information Center

    Texas Architect, 1998

    1998-01-01

    Presents selected winners of the Texas 1997 Exhibit of School Architecture Design Competition. The Caudill and honor award winning projects are listed along with facility photos, brief descriptions, project credits, and the names of the construction companies used. (GR)

  18. Exhibit of School Architecture, 1996. Special Section.

    ERIC Educational Resources Information Center

    Texas Architect, 1997

    1997-01-01

    Presents selected winners of the Texas 1996 Exhibit of School Architecture Design Competition. The Caudill and honor award-winning projects are listed along with facility photos, brief descriptions, project credits, and the names of the construction companies used. (GR)

  19. The Making of a Museum Exhibition.

    ERIC Educational Resources Information Center

    Bleecker, Samuel E.

    1979-01-01

    Discusses the preparation of the Reptile and Amphibian exhibition at the American Museum of Natural History. Various steps involved in developing the ten showcases in a six-year period are presented. (SA)

  20. 18 CFR 32.2 - Required exhibits.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... of operating such facilities. Exhibit B. A general or key map on a scale not greater than 20 miles to... facilities used for the generation and transmission of electric energy, indicating on said map the...

  1. 18 CFR 32.2 - Required exhibits.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... of operating such facilities. Exhibit B. A general or key map on a scale not greater than 20 miles to... facilities used for the generation and transmission of electric energy, indicating on said map the...

  2. 18 CFR 32.2 - Required exhibits.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... of operating such facilities. Exhibit B. A general or key map on a scale not greater than 20 miles to... facilities used for the generation and transmission of electric energy, indicating on said map the...

  3. 28 CFR 5.201 - Exhibits.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... the existing or proposed activities engaged in or to be engaged in, including political activities, by... be filed as exhibit C: (1) A copy of the registrant's charter, articles of incorporation...

  4. 28 CFR 5.201 - Exhibits.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... the existing or proposed activities engaged in or to be engaged in, including political activities, by... be filed as exhibit C: (1) A copy of the registrant's charter, articles of incorporation...

  5. 32 CFR 705.24 - Exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... public of the Navy's mission and operations. (2) To disseminate technical and scientific information. (3... naval equipment, models, devices and information and orientation material placed on public display for information purposes before audiences at conventions, conferences, seminars, demonstrations, exhibits,...

  6. Communicating Complex Sciences by Means of Exhibitions

    NASA Astrophysics Data System (ADS)

    Schneider, S.

    2011-12-01

    Earth Sciences will have to take over the leading role in global sustainable policy and in discussions about climate change. Efforts to raise attention within the politically responsible communities as well as in the public are getting more and more support by executive and advisory boards all over the world. But how can you successfully communicate complex sciences? For example, to start communication about climate change, the first step is to encourage people to be concerned about climate change. After that, one has to start thinking about how to present data and how to include the presented data into an unprejudiced context. Therefore, the communication toolbox offers various methods to reach diverse audiences. The R&D programme GEOTECHNOLOGIEN conducts roving exhibitions as one of its most successful communication tools. With roving exhibitions GEOTECHNOLOGIEN is able to get in touch with different audiences at once. The main purpose and theme of these exhibitions is to convey the everyday means of climate change to the visitors. It is within the responsibility of science to communicate the effects of a phenomenon like climate change as well as the impact of research results to the everyday life of people. Currently, a GEOTECHNOLOGIEN roving exhibition on remote sensing with satellites deals with various issues of environmental research, including a chapter on climate change. By following the 3M-concept (Meaning - Memorable - Moving), exhibitions allow to connect the visitors daily environment and personal experiences with the presented issues and objects. Therefore, hands-on exhibits, exciting multimedia effects and high-tech artefacts have to be combined with interpretive text elements to highlight the daily significance of the scientific topics and the exhibition theme respectively. To create such an exhibition, strong conceptual planning has to be conducted. This includes the specification of stern financial as well as time wise milestones. In addition

  7. When Do Children Exhibit a "Yes" Bias?

    ERIC Educational Resources Information Center

    Okanda, Mako; Itakura, Shoji

    2010-01-01

    This study investigated whether one hundred and thirty-five 3- to 6-year-old children exhibit a yes bias to various yes-no questions and whether their knowledge status affects the production of a yes bias. Three-year-olds exhibited a yes bias to all yes-no questions such as "preference-object" and "knowledge-object" questions pertaining to…

  8. An Astrobiology Microbes Exhibit and Education Module

    NASA Technical Reports Server (NTRS)

    Lindstrom, Marilyn M.; Allen, Jaclyn S.; Stocco, Karen; Tobola, Kay; Olendzenski, Lorraine

    2001-01-01

    Telling the story of NASA-sponsored scientific research to the public in exhibits is best done by partnerships of scientists and museum professionals. Likewise, preparing classroom activities and training teachers to use them should be done by teams of teachers and scientists. Here we describe how we used such partnerships to develop a new astrobiology augmentation to the Microbes! traveling exhibit and a companion education module. "Additional information is contained in the original extended abstract."

  9. Reaching the Public through Traveling Exhibitions

    NASA Astrophysics Data System (ADS)

    Dusenbery, P. B.; Harold, J. B.; Morrow, C. A.

    2004-11-01

    The Space Science Institute (SSI) of Boulder, Colorado has recently developed two museum exhibits called Alien Earths and MarsQuest. It has just started to develop another exhibit called Giant Planets. These exhibitions provide research scientists the opportunity to engage in a number of activities that are vital to the success of these major outreach programs. Alien Earths was developed in partnership with various research missions. The focus of the presentation will be on MarsQuest and Giant Planets. MarsQuest is a 5000 square-foot, \\$3M, traveling exhibition that is now touring the country. The exhibit's second 3-year tour will enable millions of Americans to share in the excitement of the scientific exploration of Mars and learn more about their own planet in the process. The associated planetarium show and education program will also be described, with particular emphasis on workshops to orient museum staff (e.g. museum educators and docents) and workshops for master educators near host museums and science centers. The workshops make innovative connections between the exhibition's interactive experiences and lesson plans aligned with the National Science Education Standards. These exhibit programs are good models for actively involving scientists and their discoveries to help improve informal science education in the museum community and for forging a stronger connection between formal and informal education. The presentation will also discuss how Giant Planets, a proposed 3500 square-foot traveling exhibition on the mysteries and discoveries of the outer planets, will be able to take advantage of the connections and resources that have been developed by the MarsQuest project.

  10. [All-Russian hygienic exhibitions and museums].

    PubMed

    Kuzybaeva, M P

    2011-01-01

    The material about the popularization of hygiene and health education in Russia in the second half of the 19th century to early 20th century through exhibition and museum activities has been collected for the first time and analyzed in the paper. The role of scientists and scientific medical societies in this process is noted. The significance of museum and exhibition activities in this area for the development of medical science is defined.

  11. Sex differences in science museum exhibit attraction

    NASA Astrophysics Data System (ADS)

    Arámbula Greenfield, Teresa

    This study examines the relative attraction of hands-on, interactive science museum exhibits for females and males. Studies have demonstrated that such exhibits can be effective learning experiences for children, with both academic and affective benefits. Other studies have shown that girls and boys do not always experience the same science-related educational opportunities and that, even when they do, they do not necessarily receive the same benefits from them. These early differences can lead to more serious educational and professional disparities later in life. As interactive museum exhibits represent a science experience that is-readily available to both girls and boys, the question arose as to whether they were being used similarly by the two groups as well as by adult women and men. It was found that both girls and boys used all types of exhibits, but that girls were more likely than boys to use puzzles and exhibits focusing on the human body; boys were more likely than girls to use computers and exhibits illustrating physical science principles. However, this was less true of children accompanied by adults (parents) than it was of unaccompanied children on school field trips who roamed the museum more freely.Received: 16 February 1994; Revised: 3 February 1995;

  12. Using Comparative Planetology in Exhibit Development

    NASA Astrophysics Data System (ADS)

    Dusenbery, P. B.; Harold, J. B.; Morrow, C. A.

    2004-12-01

    It is critically important for the public to better understand the scientific process. Museum exhibitions are an important part of informal science education that can effectively reach public audiences as well as school groups. They provide an important gateway for the public to learn about compelling scientific endeavors. The Space Science Institute (SSI) is a national leader in producing traveling science exhibitions and their associated educational programming (i.e. interactive websites, educator workshops, public talks, instructional materials). The focus of this presentation will be on three of its exhibit projects: MarsQuest (currently on tour), Alien Earths (in fabrication), and Giant Planets (in development). MarsQuest is enabling millions of Americans to share in the excitement of the scientific exploration of Mars and to learn more about their own planet in the process. Alien Earths will bring origins-related research and discoveries to students and the American public. It has four interrelated exhibit areas: Our Place in Space, Star Birth, PlanetQuest, and Search for Life. Exhibit visitors will explore the awesome events surrounding the birth of stars and planets; they will join scientists in the hunt for planets outside our solar system including those that may be in "habitable zones" around other stars; and finally they will be able to learn about how scientists are looking for signs of life beyond Earth. Giant Planets: Exploring the Outer Solar System will take advantage of the excitement generated by the Cassini mission and bring planetary and origins research and discoveries to students and the public. It will be organized around four thematic areas: Our Solar System; Colossal Worlds; Moons, Rings, and Fields; and Make Space for Kids. Giant Planets will open in 2007. This talk will focus on the importance of making Earth comparisons in the conceptual design of each exhibit and will show several examples of how these comparisons were manifested in

  13. Genome Evolution at the Genus Level: Comparison of Three Complete Genomes of Hyperthermophilic Archaea

    PubMed Central

    Lecompte, Odile; Ripp, Raymond; Puzos-Barbe, Valérie; Duprat, Simone; Heilig, Roland; Dietrich, Jacques; Thierry, Jean-Claude; Poch, Olivier

    2001-01-01

    We have compared three complete genomes of closely related hyperthermophilic species of Archaea belonging to the Pyrococcus genus: Pyrococcus abyssi, Pyrococcus horikoshii, and Pyrococcus furiosus. At the genomic level, the comparison reveals a differential conservation among four regions of the Pyrococcus chromosomes correlated with the location of genetic elements mediating DNA reorganization. This discloses the relative contribution of the major mechanisms that promote genomic plasticity in these Archaea, namely rearrangements linked to the replication terminus, insertion sequence-mediated recombinations, and DNA integration within tRNA genes. The combination of these mechanisms leads to a high level of genomic plasticity in these hyperthermophilic Archaea, at least comparable to the plasticity observed between closely related bacteria. At the proteomic level, the comparison of the three Pyrococcus species sheds light on specific selection pressures acting both on their coding capacities and evolutionary rates. Indeed, thanks to two independent methods, the “reciprocal best hits“ approach and a new distance ratio analysis, we detect the false orthology relationships within the Pyrococcus lineage. This reveals a high amount of differential gains and losses of genes since the divergence of the three closely related species. The resulting polymorphism is probably linked to an adaptation of these free-living organisms to differential environmental constraints. As a corollary, we delineate the set of orthologous genes shared by the three species, that is, the genes that may characterize the Pyrococcus genus. In this conserved core, the amino acid substitution rate is equal between P. abyssi and P. horikoshii for most of their shared proteins, even for fast-evolving ones. In contrast, strong discrepancies exist among the substitution rates observed in P. furiosus relative to the two other species, which is in disagreement with the molecular clock hypothesis. PMID

  14. After Terror Charges, Artist Exhibits Academic Freedom

    ERIC Educational Resources Information Center

    Wilson, Robin

    2008-01-01

    Steven Kurtz, a professor of visual studies at the State University of New York, has been working with various bacteria as part of his counterculture exhibit artworks for nearly 20 years. Four years ago, federal agents raided his home in a bioterrorism investigation. The federal agents had been called to the house by local police officers…

  15. The medial prefrontal cortex exhibits money illusion

    PubMed Central

    Weber, Bernd; Rangel, Antonio; Wibral, Matthias; Falk, Armin

    2009-01-01

    Behavioral economists have proposed that money illusion, which is a deviation from rationality in which individuals engage in nominal evaluation, can explain a wide range of important economic and social phenomena. This proposition stands in sharp contrast to the standard economic assumption of rationality that requires individuals to judge the value of money only on the basis of the bundle of goods that it can buy—its real value—and not on the basis of the actual amount of currency—its nominal value. We used fMRI to investigate whether the brain's reward circuitry exhibits money illusion. Subjects received prizes in 2 different experimental conditions that were identical in real economic terms, but differed in nominal terms. Thus, in the absence of money illusion there should be no differences in activation in reward-related brain areas. In contrast, we found that areas of the ventromedial prefrontal cortex (vmPFC), which have been previously associated with the processing of anticipatory and experienced rewards, and the valuation of goods, exhibited money illusion. We also found that the amount of money illusion exhibited by the vmPFC was correlated with the amount of money illusion exhibited in the evaluation of economic transactions. PMID:19307555

  16. 18 CFR 153.8 - Required exhibits.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... LNG, evidence that an appropriate and qualified concern will properly and safely receive or deliver such LNG, including a report containing detailed engineering and design information. The Commission... Office of Energy Projects, 888 First Street, NE., Washington, DC 20426; (6) Exhibit E-1. If the...

  17. 28 CFR 68.43 - Exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 28 Judicial Administration 2 2010-07-01 2010-07-01 false Exhibits. 68.43 Section 68.43 Judicial Administration DEPARTMENT OF JUSTICE (CONTINUED) RULES OF PRACTICE AND PROCEDURE FOR ADMINISTRATIVE HEARINGS BEFORE ADMINISTRATIVE LAW JUDGES IN CASES INVOLVING ALLEGATIONS OF UNLAWFUL EMPLOYMENT OF ALIENS,...

  18. Creating Cross-Cultural Exhibits in Schools.

    ERIC Educational Resources Information Center

    Nakamura, Kazuyo; Erickson, Virginia; Ford, Viktoria

    Theory and practice of the Cross-Cultural Arts Exhibit project initiated by the Krannert Art Museum at the University of Illinois at Urbana-Champaign (Illinois) is described in this paper. The project was developed based on the concept of post-museum. Instead of transmitting values and knowledge, communication in the post-museum stresses the…

  19. 18 CFR 50.7 - Applications: exhibits.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... required supporting equipment, such as insulation medium pressurizing or forced cooling; (iii) Cathodic protection scheme; and (iv) Type of dielectric fluid and safeguards used to limit potential spills in...) Exhibit H—System analysis data. An analysis evaluating the impact the proposed facilities will have on...

  20. 17 CFR 232.102 - Exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... pursuant to the schedule's general instructions. See Rule 311(b) of Regulation S-T (17 CFR 232.311(b)). (b... registered investment company or a business development company). (d) Each electronic filing requiring... AND REGULATIONS FOR ELECTRONIC FILINGS Electronic Filing Requirements § 232.102 Exhibits. (a)...

  1. 18 CFR 50.7 - Applications: exhibits.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 18 Conservation of Power and Water Resources 1 2011-04-01 2011-04-01 false Applications: exhibits. 50.7 Section 50.7 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION, DEPARTMENT OF ENERGY REGULATIONS UNDER THE FEDERAL POWER ACT APPLICATIONS FOR PERMITS TO SITE...

  2. 18 CFR 50.7 - Applications: exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... relation to the project and other principal interconnected system elements, as well as power flow and loss... 18 Conservation of Power and Water Resources 1 2010-04-01 2010-04-01 false Applications: exhibits. 50.7 Section 50.7 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY...

  3. 18 CFR 50.7 - Applications: exhibits.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 18 Conservation of Power and Water Resources 1 2012-04-01 2012-04-01 false Applications: exhibits. 50.7 Section 50.7 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION... destination of the project; (ii) Design voltage rating (kV); (iii) Operating voltage rating (kV); (iv)...

  4. The medial prefrontal cortex exhibits money illusion.

    PubMed

    Weber, Bernd; Rangel, Antonio; Wibral, Matthias; Falk, Armin

    2009-03-31

    Behavioral economists have proposed that money illusion, which is a deviation from rationality in which individuals engage in nominal evaluation, can explain a wide range of important economic and social phenomena. This proposition stands in sharp contrast to the standard economic assumption of rationality that requires individuals to judge the value of money only on the basis of the bundle of goods that it can buy-its real value-and not on the basis of the actual amount of currency-its nominal value. We used fMRI to investigate whether the brain's reward circuitry exhibits money illusion. Subjects received prizes in 2 different experimental conditions that were identical in real economic terms, but differed in nominal terms. Thus, in the absence of money illusion there should be no differences in activation in reward-related brain areas. In contrast, we found that areas of the ventromedial prefrontal cortex (vmPFC), which have been previously associated with the processing of anticipatory and experienced rewards, and the valuation of goods, exhibited money illusion. We also found that the amount of money illusion exhibited by the vmPFC was correlated with the amount of money illusion exhibited in the evaluation of economic transactions.

  5. MarsQuest: A National Traveling Exhibition

    NASA Astrophysics Data System (ADS)

    Lee, S. W.; Dusenbery, P. B.

    1998-09-01

    With the successful landing of Mars Pathfinder and the arrival of Mars Global Surveyor, a new decade of Mars exploration has commenced. MarsQuest, a 5000 square foot traveling exhibition, is being developed to further bring the excitement and discoveries of this "Decade of Mars Exploration" to the public. MarsQuest is partially funded by the Informal Science Education Program of the National Science Foundation and NASA's Office of Space Science. The Space Science Institute (SSI) in Boulder, CO, is leading the project. Scientific and educational advisors from many different universities and government laboratories, most of whom are directly involved in the active and planned Mars missions, will ensure the scientific accuracy, timeliness, and relevance of the key concepts presented in the exhibition and accompanying programs. The traveling exhibit is the primary element of the MarsQuest project. The exhibition experience, carefully keyed to current events in Mars exploration, will transport visitors to the surface of the Red Planet via large murals, dioramas, and numerous interactive displays. There they will have the opportunity to share in the spirit and thrill of exploration, and come to appreciate the similarities and differences between Earth and Mars. A planetarium show, geared to the goals of the MarsQuest project, will be an important sensory addition to the traveling exhibit. The planetarium/star-theater venue presents a unique environment where audience members can literally be surrounded by Mars images. Education and outreach programs comprise the remainder of the MarsQuest project. The goal of these is to make scientific concepts and scientific and engineering processes understandable to students via Mars-inspired curricula. MarsQuest will open in late-1999, traveling to about nine sites throughout the United States and reaching an estimated two to three million children and adults during its planned three-year tour. Mars - coming soon to a museum near

  6. The palaeontological exhibition: a venue for dialogue.

    PubMed

    Murriello, Sandra

    2015-01-01

    Understanding the dialogue between museums and their visitors enables museums to subsist, undergo transformations and become consolidated as socially valued cultural venues. The Museo de La Plata (Argentina) was created in the late nineteenth century as a natural history museum, and this study shows that currently the museum is valued socially as a venue for family leisure and education, at which people make sense to the objects exhibited through characteristics conferred upon them by both the institution and the visitor. Nevertheless, such dialogue is somehow affected by the museographic proposal and the public interpretation of the institutional narrative, which could be analysed within the frame of contextual learning. As a consequence, the evolutionary idea that the museum aims to communicate is distorted by the public. This article highlights the importance of considering the visitors' interpretations when planning museum exhibitions, a perspective that has been rather absent in the Argentinian museums.

  7. Multimodal audio guide for museums and exhibitions

    NASA Astrophysics Data System (ADS)

    Gebbensleben, Sandra; Dittmann, Jana; Vielhauer, Claus

    2006-02-01

    In our paper we introduce a new Audio Guide concept for exploring buildings, realms and exhibitions. Actual proposed solutions work in most cases with pre-defined devices, which users have to buy or borrow. These systems often go along with complex technical installations and require a great degree of user training for device handling. Furthermore, the activation of audio commentary related to the exhibition objects is typically based on additional components like infrared, radio frequency or GPS technology. Beside the necessity of installation of specific devices for user location, these approaches often only support automatic activation with no or limited user interaction. Therefore, elaboration of alternative concepts appears worthwhile. Motivated by these aspects, we introduce a new concept based on usage of the visitor's own mobile smart phone. The advantages in our approach are twofold: firstly the Audio Guide can be used in various places without any purchase and extensive installation of additional components in or around the exhibition object. Secondly, the visitors can experience the exhibition on individual tours only by uploading the Audio Guide at a single point of entry, the Audio Guide Service Counter, and keeping it on her or his personal device. Furthermore, since the user usually is quite familiar with the interface of her or his phone and can thus interact with the application device easily. Our technical concept makes use of two general ideas for location detection and activation. Firstly, we suggest an enhanced interactive number based activation by exploiting the visual capabilities of modern smart phones and secondly we outline an active digital audio watermarking approach, where information about objects are transmitted via an analog audio channel.

  8. Naval Meteorology and Oceanography Command exhibit entrance

    NASA Technical Reports Server (NTRS)

    2000-01-01

    StenniSphere at NASA's John C. Stennis Space Center in Hancock County, Miss., invites visitors to discover why America comes to Stennis Space Center before going into space. Designed to entertain while educating, StenniSphere includes informative displays and exhibits from NASA and other agencies located at Stennis, such as this one from the Naval Meteorology and Oceanography Command. Visitors can 'travel' three-dimensionally under the sea and check on the weather back home in the Weather Center.

  9. Naval Meteorology and Oceanography Command exhibit

    NASA Technical Reports Server (NTRS)

    2000-01-01

    Designed to entertain while educating, StenniSphere at the John C. Stennis Space Center in Hancock County, Miss., includes informative displays and exhibits from NASA and other agencies located at Stennis, such as this one from the Naval Meteorology and Oceanography Command. Visitors can 'travel' three-dimensionally under the sea and check on the weather back home in the Weather Center. StenniSphere is open free of charge from 9 a.m. to 5 p.m. daily.

  10. Art exhibit focuses on African astronomy

    NASA Astrophysics Data System (ADS)

    Showstack, Randy

    2012-07-01

    Connections between Africans and astronomy are the focus of a new exhibition in the National Museum of African Art in Washington, D. C. "African Cosmos: Stellar Arts," which includes artwork, cultural items, and scientific displays from ancient to contemporary times, is the first major exhibit "that brings together arts and science focused on Africa's contribution to keen observations of the heavens over time," curator Christine Mullen Kreamer said at a 20 June news briefing. Among the exhibit's nearly 100 objects are an ancient Egyptian mummy board that includes a representation of the sky goddess Nut, sculptures by the Dogon people of Mali depicting figures in relation to the cosmos, a video that uses data from two square degrees of the Hubble Space Telescope Cosmic Evolution Survey, and a nearly floor-to-ceiling "Rainbow Serpent" constructed of plastic containers by Benin artist Hazoume. An untitled acrylic painting (Figure 1) by South African Gavin Jantjes evokes a myth of the Khoi San people of southern Africa, as it portrays a girl throwing evening fire embers into the night sky, where they remained as the Milky Way.

  11. Bumblebees exhibit the memory spacing effect

    NASA Astrophysics Data System (ADS)

    Toda, Nicholas R. T.; Song, Jeremy; Nieh, James C.

    2009-10-01

    Associative learning is key to how bees recognize and return to rewarding floral resources. It thus plays a major role in pollinator floral constancy and plant gene flow. Honeybees are the primary model for pollinator associative learning, but bumblebees play an important ecological role in a wider range of habitats, and their associative learning abilities are less well understood. We assayed learning with the proboscis extension reflex (PER), using a novel method for restraining bees (capsules) designed to improve bumblebee learning. We present the first results demonstrating that bumblebees exhibit the memory spacing effect. They improve their associative learning of odor and nectar reward by exhibiting increased memory acquisition, a component of long-term memory formation, when the time interval between rewarding trials is increased. Bombus impatiens forager memory acquisition (average discrimination index values) improved by 129% and 65% at inter-trial intervals (ITI) of 5 and 3 min, respectively, as compared to an ITI of 1 min. Memory acquisition rate also increased with increasing ITI. Encapsulation significantly increases olfactory memory acquisition. Ten times more foragers exhibited at least one PER response during training in capsules as compared to traditional PER harnesses. Thus, a novel conditioning assay, encapsulation, enabled us to improve bumblebee-learning acquisition and demonstrate that spaced learning results in better memory consolidation. Such spaced learning likely plays a role in forming long-term memories of rewarding floral resources.

  12. When do children exhibit a "yes" bias?

    PubMed

    Okanda, Mako; Itakura, Shoji

    2010-01-01

    This study investigated whether one hundred and thirty-five 3- to 6-year-old children exhibit a yes bias to various yes-no questions and whether their knowledge status affects the production of a yes bias. Three-year-olds exhibited a yes bias to all yes-no questions such as preference-object and knowledge-object questions pertaining to objects, and knowledge-face questions pertaining to facial expressions. Four-year-olds tended to say "yes" only to knowledge-object questions. Five-year-olds did not show any strong response tendency. Six-year-olds exhibited a nay-saying bias to knowledge-face questions. Also, 3-year-olds could indicate the correct option when asked questions with 2 response options. It suggested that 3-year-olds tended to inappropriately say "yes" to yes-no questions, although they knew the answers to the questions. The mechanism of a yes bias was discussed.

  13. The E = mc{sup 2} exhibition

    SciTech Connect

    Henderson, D.; Peshkin, M.

    1995-08-01

    The goal of this DOE-supported exhibition is to demystify Einstein`s formula E = mc{sup 2} by illustrating the interchangeability of matter (m) and energy (E), c{sup 2} being the exchange rate. The exhibition has two major parts, {open_quotes}matter into energy{close_quotes} and {open_quotes}energy into matter{close_quotes}, plus a video to connect them. {open_quotes}Matter into energy{close_quotes} has now been completed and has been placed on the museum floor. Positrons from a {sup 22}Na source are annihilated to produce gamma rays that are caught in NaI detectors. The viewer can alter the alignment of the detectors and observe the consequences for the rates of single and coincident counts. The viewer can also observe the effects of placing absorbers in front of the counters. Prototype explanatory graphics were placed around the exhibit and those will probably be changed after we have some experience with their effectiveness. The connecting video is in the process of being produced in collaboration with Fermilab. A cloud chamber for {open_quotes}energy into matter{close_quotes}, where gamma rays from a small Th source will produce observable pairs, was purchased and work to make the pairs visible has commenced.

  14. 7 CFR Exhibit G to Subpart E of... - Exhibit G to Subpart E of Part 1980

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... OF AGRICULTURE (CONTINUED) PROGRAM REGULATIONS (CONTINUED) GENERAL Business and Industrial Loan... 7 Agriculture 14 2014-01-01 2014-01-01 false Exhibit G to Subpart E of Part 1980 G Exhibit G to Subpart E of Part 1980 Agriculture Regulations of the Department of Agriculture (Continued) RURAL...

  15. 7 CFR Exhibit G to Subpart E of... - Exhibit G to Subpart E of Part 1980

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... OF AGRICULTURE (CONTINUED) PROGRAM REGULATIONS (CONTINUED) GENERAL Business and Industrial Loan... 7 Agriculture 14 2011-01-01 2011-01-01 false Exhibit G to Subpart E of Part 1980 G Exhibit G to Subpart E of Part 1980 Agriculture Regulations of the Department of Agriculture (Continued) RURAL...

  16. 7 CFR Exhibit G to Subpart E of... - Exhibit G to Subpart E of Part 1980

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... OF AGRICULTURE (CONTINUED) PROGRAM REGULATIONS (CONTINUED) GENERAL Business and Industrial Loan... 7 Agriculture 14 2012-01-01 2012-01-01 false Exhibit G to Subpart E of Part 1980 G Exhibit G to Subpart E of Part 1980 Agriculture Regulations of the Department of Agriculture (Continued) RURAL...

  17. 7 CFR Exhibit G to Subpart E of... - Exhibit G to Subpart E of Part 1980

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... OF AGRICULTURE (CONTINUED) PROGRAM REGULATIONS (CONTINUED) GENERAL Business and Industrial Loan... 7 Agriculture 14 2010-01-01 2009-01-01 true Exhibit G to Subpart E of Part 1980 G Exhibit G to Subpart E of Part 1980 Agriculture Regulations of the Department of Agriculture (Continued) RURAL...

  18. 7 CFR Exhibit G to Subpart E of... - Exhibit G to Subpart E of Part 1980

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... OF AGRICULTURE (CONTINUED) PROGRAM REGULATIONS (CONTINUED) GENERAL Business and Industrial Loan... 7 Agriculture 14 2013-01-01 2013-01-01 false Exhibit G to Subpart E of Part 1980 G Exhibit G to Subpart E of Part 1980 Agriculture Regulations of the Department of Agriculture (Continued) RURAL...

  19. Mars in their eyes - a cartoon exhibition

    NASA Astrophysics Data System (ADS)

    Pillinger, Pi.

    Recently a collection of 120 cartoons which tell the story of Mars exploration and scientific discovery, past, present and future, was held in London. We discuss the aims of the exhibition, to what extent we believe the original aims were met and report on additional outreach opportunities resulting from the project. The overriding aim was to capitalise on the popular appeal of accessible art - most people admit to enjoying cartoons. This was strengthened by hanging the originals of cartoons which had, mostly, been published in newspapers and magazines in a wide selection of countries. The provenances served to indicate the attraction of Mars to a wide public. We were fortunate to work with the Cartoon Art Trust of the UK who was in the process of relocating to new premises and opening as The Cartoon Museum, in the tourist area of Bloomsbury, central London, very close to the British Museum. "Mars in their Eyes" ran for 10 weeks during April to July 2006; immediately following which a selection of the cartoons was displayed at the week-long Royal Society Summer Exhibition. We explore the differences between the two exhibitions and comment on the various audience responses. We use this comparison to discuss whether a project which is primarily art can be extended to explain science. Does the coupling merely result in dumbing-down of both cultures or is there a true synergy? The experience has led us to coin the phrase "extreme outreach". Projects which are as ambitious as "Mars in their Eyes", without the security of a safe, captive audience, for example at a Science Centre, must be judged by different criteria. Indeed if the project does not meet comparable targets like large visitor numbers, then the honest evaluation of such details can only inform future activities and must not be reflected in the future funding of only "safe" outreach activities.

  20. Noisy neural nets exhibiting epileptic features.

    PubMed

    Kokkinidis, M; Anninos, P

    1985-04-01

    On the basis of our previous studies of noisy neural nets we propose a model for the explanation of epileptic phenomena. Our neural net model is capable of exhibiting epileptic features if the number of spontaneously firing neurons is periodically increased beyond a certain threshold. Some alternative epileptogenic mechanisms are also discussed. The epileptic behavior of the neural net is determined by a combination of certain parameters of its phase diagram. The general features of the model are consistent with several experimental observations and explain some poorly understood clinical phenomena. The differences between normal and epileptic neural nets are explained in terms of the structural properties of the model.

  1. Library exhibits and programs boost science education

    NASA Astrophysics Data System (ADS)

    Dusenbery, Paul B.; Curtis, Lisa

    2012-05-01

    Science museums let visitors explore and discover, but for many families there are barriers—such as cost or distance—that prevent them from visiting museums and experiencing hands-on science, technology, engineering, and mathematics (STEM) learning. Now educators are reaching underserved audiences by developing STEM exhibits and programs for public libraries. With more than 16,000 outlets in the United States, public libraries serve almost every community in the country. Nationwide, they receive about 1.5 billion visits per year, and they offer their services for free.

  2. Nematic liquid crystals exhibiting high birefringence

    NASA Astrophysics Data System (ADS)

    Thingujam, Kiranmala; Bhattacharjee, Ayon; Choudhury, Basana; Dabrowski, Roman

    2016-06-01

    Two fluorinated isothiocyanato nematic liquid crystalline compounds, 4'-butylcyclohexyl-3, 5-difluoro-4-isothiocyanatobiphenyl and 4'-pentylcyclohexyl-3, 5-difluoro-4-isothiocynatobiphenyl are studied in detail to obtain their different physical parameters. Optical polarizing microscopy, differential scanning calorimetry, density and dielectric studies have been carried out for the two samples. Both the samples were found to have high clearing temperature (>100 °C) and exhibit small enthalpy of transition. The two samples exhibit high optical birefringence (Δ n > 0.2). The values of order parameters for the two samples were obtained using different approaches, namely, Vuks', Neugebauer's, modified Vuks' and direct extrapolation method from birefringence data. Experimentally obtained values of order parameters have also been compared with theoretical Maier-Saupe values. The parallel and perpendicular components of dielectric permittivity values of the two compounds were also calculated and their anisotropy values were found to be small. The effect of temperature on the molecular dipole moment μ and the angle of inclination β of the dipole axis with the director have also been investigated in this work.

  3. Supercomputing meets seismology in earthquake exhibit

    SciTech Connect

    Blackwell, Matt; Rodger, Arthur; Kennedy, Tom

    2013-10-03

    When the California Academy of Sciences created the "Earthquake: Evidence of a Restless Planet" exhibit, they called on Lawrence Livermore to help combine seismic research with the latest data-driven visualization techniques. The outcome is a series of striking visualizations of earthquakes, tsunamis and tectonic plate evolution. Seismic-wave research is a core competency at Livermore. While most often associated with earthquakes, the research has many other applications of national interest, such as nuclear explosion monitoring, explosion forensics, energy exploration, and seismic acoustics. For the Academy effort, Livermore researchers simulated the San Andreas and Hayward fault events at high resolutions. Such calculations require significant computational resources. To simulate the 1906 earthquake, for instance, visualizing 125 seconds of ground motion required over 1 billion grid points, 10,000 time steps, and 7.5 hours of processor time on 2,048 cores of Livermore's Sierra machine.

  4. New Monolayered Materials Exhibiting Unusual Electronic Properties

    NASA Astrophysics Data System (ADS)

    Lopez-Bezanilla, Alejandro; Martin, Ivar; Littlewood, Peter B.

    Computationally based approaches are allowing to progress in the discovery and design of nano-scaled materials. Here we propose a series of new mono-layered compounds with exotic properties. By means of density functional theory calculations we demonstrate that the pentagonal arrangement of SiC2 yields an inverted distribution of the p-bands which leads to an unusual electronic behaviour of the material under strain [J. Phys. Chem. C, 2015, 119 (33), pp 19469]. A different pentagonal arrangement of C atoms enables the formation of Dirac cones which, unlike graphene, exhibit a strain-mediated tunable band gap. This work is supported by DOE-BES under Contract No. DE-AC02-06CH11357.

  5. Supercomputing meets seismology in earthquake exhibit

    ScienceCinema

    Blackwell, Matt; Rodger, Arthur; Kennedy, Tom

    2016-07-12

    When the California Academy of Sciences created the "Earthquake: Evidence of a Restless Planet" exhibit, they called on Lawrence Livermore to help combine seismic research with the latest data-driven visualization techniques. The outcome is a series of striking visualizations of earthquakes, tsunamis and tectonic plate evolution. Seismic-wave research is a core competency at Livermore. While most often associated with earthquakes, the research has many other applications of national interest, such as nuclear explosion monitoring, explosion forensics, energy exploration, and seismic acoustics. For the Academy effort, Livermore researchers simulated the San Andreas and Hayward fault events at high resolutions. Such calculations require significant computational resources. To simulate the 1906 earthquake, for instance, visualizing 125 seconds of ground motion required over 1 billion grid points, 10,000 time steps, and 7.5 hours of processor time on 2,048 cores of Livermore's Sierra machine.

  6. Application of an imaging system to a museum exhibition for developing interactive exhibitions

    NASA Astrophysics Data System (ADS)

    Miyata, Kimiyoshi; Inoue, Yuka; Takiguchi, Takahiro; Tsumura, Norimichi; Nakaguchi, Toshiya; Miyake, Yoichi

    2009-10-01

    In the National Museum of Japanese History, 215,759 artifacts are stored and used for research and exhibitions. In museums, due to the limitation of space in the galleries, a guidance system is required to satisfy visitors' needs and to enhance their understanding of the artifacts. We introduce one exhibition using imaging technology to improve visitors' understanding of a kimono (traditional Japanese clothing) exhibition. In the imaging technology introduced, one data projector, one display with touch panel interface, and magnifiers were used as exhibition tools together with a real kimono. The validity of this exhibition method was confirmed by results from a visitors' interview survey. Second, to further develop the interactive guidance system, an augmented reality system that consisted of cooperation between the projector and a digital video camera was also examined. A white paper board in the observer's hand was used as a projection screen and also as an interface to control the images projected on the board. The basic performance of the proposed system was confirmed; however continuous development was necessary for applying the system to actual exhibitions.

  7. A Traveling Exhibit of Cassini Image Science

    NASA Astrophysics Data System (ADS)

    Burns, Joseph A.; Hedman, M. M.; Tiscareno, M. S.; Ebel, D.; Mac Low, M.; Lovett, L. E.; Burns, J. K.; Schaff, N.; Bilson, E. M.

    2007-10-01

    An exhibit of Cassini's images will open at NYC's American Museum of Natural History in March 2008 and then visit the Johnson Art Museum (Cornell) throughout fall 2008, including during next year's DPS. It is under consideration by several other venues in the States and overseas. The exhibit will feature 40-50 images, ranging from letter size to large posters, taken by remote-sensing instruments aboard Cassini and Huygens. Photos will be organized into a half-dozen thematic clusters (e.g., organized by celestial target or by physical process); a panel will introduce each grouping with individual images identified briefly. The Saturn system is a perfect vehicle to educate citizens about planetary science and origins. The images’ beauty should capture the public's attention, allowing us to then engage their curiosity about the relevant science. Among the Saturn system's broad suite of objects are Enceladus and Titan, two satellites of astrobiological interest; moreover, the rings display many processes active in other astrophysical disks. Several auxiliary ideas will be implemented. In Ithaca, we will project images at night against the museum's sand-colored exterior walls. A 10-12 minute musical composition has been commissioned from Roberto Sierra to open the show. We will encourage school children to participate in a human orrery circling the museum and will seek volunteers to participate in several Saturnalia. At Cornell we will involve the university and local communities, by taping their reactions to the images’ exquisite beauty as well as to their scientific content. Cassini will be the E/PO focus of next year's DPS meeting; those materials will be employed throughout the fall at New York schools and be available to travel with the show. We intend to work with NYC partners to offer teacher credits for associated weekend courses. We will produce classroom materials, including a DVD, for teacher use.

  8. Quiescent Fibroblasts Exhibit High Metabolic Activity

    PubMed Central

    Lemons, Johanna M. S.; Feng, Xiao-Jiang; Bennett, Bryson D.; Legesse-Miller, Aster; Johnson, Elizabeth L.; Raitman, Irene; Pollina, Elizabeth A.; Rabitz, Herschel A.; Rabinowitz, Joshua D.; Coller, Hilary A.

    2010-01-01

    Many cells in mammals exist in the state of quiescence, which is characterized by reversible exit from the cell cycle. Quiescent cells are widely reported to exhibit reduced size, nucleotide synthesis, and metabolic activity. Much lower glycolytic rates have been reported in quiescent compared with proliferating lymphocytes. In contrast, we show here that primary human fibroblasts continue to exhibit high metabolic rates when induced into quiescence via contact inhibition. By monitoring isotope labeling through metabolic pathways and quantitatively identifying fluxes from the data, we show that contact-inhibited fibroblasts utilize glucose in all branches of central carbon metabolism at rates similar to those of proliferating cells, with greater overflow flux from the pentose phosphate pathway back to glycolysis. Inhibition of the pentose phosphate pathway resulted in apoptosis preferentially in quiescent fibroblasts. By feeding the cells labeled glutamine, we also detected a “backwards” flux in the tricarboxylic acid cycle from α-ketoglutarate to citrate that was enhanced in contact-inhibited fibroblasts; this flux likely contributes to shuttling of NADPH from the mitochondrion to cytosol for redox defense or fatty acid synthesis. The high metabolic activity of the fibroblasts was directed in part toward breakdown and resynthesis of protein and lipid, and in part toward excretion of extracellular matrix proteins. Thus, reduced metabolic activity is not a hallmark of the quiescent state. Quiescent fibroblasts, relieved of the biosynthetic requirements associated with generating progeny, direct their metabolic activity to preservation of self integrity and alternative functions beneficial to the organism as a whole. PMID:21049082

  9. Nuclear resonance vibrational spectroscopy (NRVS) of rubredoxin and MoFe protein crystals

    PubMed Central

    Guo, Yisong; Brecht, Eric; Aznavour, Kristen; Nix, Jay C.; Xiao, Yuming; Wang, Hongxin; George, Simon J.; Bau, Robert; Keable, Stephen; Peters, John W.; Adams, Michael W.W.; Jenney, Francis; Sturhahn, Wolfgang; Alp, Ercan E.; Zhao, Jiyong; Yoda, Yoshitaka; Cramer, Stephen P.

    2014-01-01

    We have applied 57Fe nuclear resonance vibrational spectroscopy (NRVS) for the first time to study the dynamics of Fe centers in Fe-S protein crystals, including oxidized wild type rubredoxin crystals from Pyrococcus furiosus, and the MoFe protein of nitrogenase from Azotobacter vinelandii. Thanks to the NRVS selection rule, selectively probed vibrational modes have been observed in both oriented rubredoxin and MoFe protein crystals. The NRVS work was complemented by extended X-ray absorption fine structure spectroscopy (EXAFS) measurements on oxidized wild type rubredoxin crystals from Pyrococcus furiosus. The EXAFS spectra revealed the Fe-S bond length difference in oxidized Pf Rd protein, which is qualitatively consistent with the X-ray crystal structure. PMID:26052177

  10. Nuclear resonance vibrational spectroscopy (NRVS) of rubredoxin and MoFe protein crystals

    NASA Astrophysics Data System (ADS)

    Guo, Yisong; Brecht, Eric; Aznavour, Kristen; Nix, Jay C.; Xiao, Yuming; Wang, Hongxin; George, Simon J.; Bau, Robert; Keable, Stephen; Peters, John W.; Adams, Michael W. W.; , Francis E. Jenney, Jr.; Sturhahn, Wolfgang; Alp, Ercan E.; Zhao, Jiyong; Yoda, Yoshitaka; Cramer, Stephen P.

    2013-12-01

    We have applied 57Fe nuclear resonance vibrational spectroscopy (NRVS) for the first time to study the dynamics of Fe centers in Iron-sulfur protein crystals, including oxidized wild type rubredoxin crystals from Pyrococcus furiosus, and the MoFe protein of nitrogenase from Azotobacter vinelandii. Thanks to the NRVS selection rule, selectively probed vibrational modes have been observed in both oriented rubredoxin and MoFe protein crystals. The NRVS work was complemented by extended X-ray absorption fine structure spectroscopy (EXAFS) measurements on oxidized wild type rubredoxin crystals from Pyrococcus furiosus. The EXAFS spectra revealed the Fe-S bond length difference in oxidized Pf Rd protein, which is qualitatively consistent with the crystal structure.

  11. Exhibition of Stochastic Resonance in Vestibular Perception

    NASA Technical Reports Server (NTRS)

    Galvan-Garza, R. C.; Clark, T. K.; Merfeld, D. M.; Bloomberg, J. J.; Oman, C. M.; Mulavara, A. P.

    2016-01-01

    Astronauts experience sensorimotor changes during spaceflight, particularly during G-transitions. Post flight sensorimotor changes include spatial disorientation, along with postural and gait instability that may degrade operational capabilities of the astronauts and endanger the crew. A sensorimotor countermeasure that mitigates these effects would improve crewmember safety and decrease risk. The goal of this research is to investigate the potential use of stochastic vestibular stimulation (SVS) as a technology to improve sensorimotor function. We hypothesize that low levels of SVS will improve sensorimotor perception through the phenomenon of stochastic resonance (SR), when the response of a nonlinear system to a weak input signal is enhanced by the application of a particular nonzero level of noise. This study aims to advance the development of SVS as a potential countermeasure by 1) demonstrating the exhibition of stochastic resonance in vestibular perception, a vital component of sensorimotor function, 2) investigating the repeatability of SR exhibition, and 3) determining the relative contribution of the semicircular canals (SCC) and otolith (OTO) organs to vestibular perceptual SR. A constant current stimulator was used to deliver bilateral bipolar SVS via electrodes placed on each of the mastoid processes, as previously done. Vestibular perceptual motion recognition thresholds were measured using a 6-degree of freedom MOOG platform and a 150 trial 3-down/1-up staircase procedure. In the first test session, we measured vestibular perceptual thresholds in upright roll-tilt at 0.2 Hz (SCC+OTO) with SVS ranging from 0-700 µA. In a second test session a week later, we re-measured roll-tilt thresholds with 0, optimal (from test session 1), and 1500 µA SVS levels. A subset of these subjects, plus naive subjects, participated in two additional test sessions in which we measured thresholds in supine roll-rotation at 0.2 Hz (SCC) and upright y-translation at 1 Hz

  12. Agitated Honeybees Exhibit Pessimistic Cognitive Biases

    PubMed Central

    Bateson, Melissa; Desire, Suzanne; Gartside, Sarah E.; Wright, Geraldine A.

    2011-01-01

    Summary Whether animals experience human-like emotions is controversial and of immense societal concern [1–3]. Because animals cannot provide subjective reports of how they feel, emotional state can only be inferred using physiological, cognitive, and behavioral measures [4–8]. In humans, negative feelings are reliably correlated with pessimistic cognitive biases, defined as the increased expectation of bad outcomes [9–11]. Recently, mammals [12–16] and birds [17–20] with poor welfare have also been found to display pessimistic-like decision making, but cognitive biases have not thus far been explored in invertebrates. Here, we ask whether honeybees display a pessimistic cognitive bias when they are subjected to an anxiety-like state induced by vigorous shaking designed to simulate a predatory attack. We show for the first time that agitated bees are more likely to classify ambiguous stimuli as predicting punishment. Shaken bees also have lower levels of hemolymph dopamine, octopamine, and serotonin. In demonstrating state-dependent modulation of categorization in bees, and thereby a cognitive component of emotion, we show that the bees' response to a negatively valenced event has more in common with that of vertebrates than previously thought. This finding reinforces the use of cognitive bias as a measure of negative emotional states across species and suggests that honeybees could be regarded as exhibiting emotions. Video Abstract PMID:21636277

  13. Ferromagnetism exhibited by nanoparticles of noble metals.

    PubMed

    Maitra, Urmimala; Das, Barun; Kumar, Nitesh; Sundaresan, Athinarayanan; Rao, C N R

    2011-08-22

    Gold nanoparticles with average diameters in the range 2.5-15 nm, prepared at the organic/aqueous interface by using tetrakis(hydroxymethyl)phosphonium chloride (THPC) as reducing agent, exhibit ferromagnetism whereby the saturation magnetization M(S) increases with decreasing diameter and varies linearly with the fraction of surface atoms. The value of M(S) is higher when the particles are present as a film instead of as a sol. Capping with strongly interacting ligands such as alkane thiols results in a higher M(S) value, which varies with the strength of the metal-sulfur bond. Ferromagnetism is also found in Pt and Ag nanoparticles prepared as sols, and the M(S) values vary as Pt>Au>Ag. A careful study of the temperature variation of the magnetization of Au nanoparticles, along with certain other observations, suggests that small bare nanoparticles of noble metals could indeed possess ferromagnetism, albeit weak, which is accentuated in the presence of capping agents, specially alkane thiols which form strong metal-sulfur bonds.

  14. Marine bacteria exhibit a bipolar distribution.

    PubMed

    Sul, Woo Jun; Oliver, Thomas A; Ducklow, Hugh W; Amaral-Zettler, Linda A; Sogin, Mitchell L

    2013-02-01

    The microbial cosmopolitan dispersion hypothesis often invoked to explain distribution patterns driven by high connectivity of oceanographic water masses and widespread dispersal ability has never been rigorously tested. By using a global marine bacterial dataset and iterative matrix randomization simulation, we show that marine bacteria exhibit a significantly greater dispersal limitation than predicted by our null model using the "everything is everywhere" tenet with no dispersal limitation scenario. Specifically, marine bacteria displayed bipolar distributions (i.e., species occurring exclusively at both poles and nowhere else) significantly less often than in the null model. Furthermore, we observed fewer taxa present in both hemispheres but more taxa present only in a single hemisphere than expected under the null model. Each of these trends diverged further from the null expectation as the compared habitats became more geographically distant but more environmentally similar. Our meta-analysis supported a latitudinal gradient in bacterial diversity with higher richness at lower latitudes, but decreased richness toward the poles. Bacteria in the tropics also demonstrated narrower latitudinal ranges at lower latitudes and relatively larger ranges in higher latitudes, conforming to the controversial macroecological pattern of the "Rapoport rule." Collectively, our findings suggest that bacteria follow biogeographic patterns more typical of macroscopic organisms, and that dispersal limitation, not just environmental selection, likely plays an important role. Distributions of microbes that deliver critical ecosystem services, particularly those in polar regions, may be vulnerable to the same impacts that environmental stressors, climate warming, and degradation in habitat quality are having on biodiversity in animal and plant species.

  15. Waves in geomaterials exhibiting negative stiffness behaviour

    NASA Astrophysics Data System (ADS)

    Esin, Maxim; Dyskin, Arcady; Pasternak, Elena

    2016-04-01

    Negative stiffness denotes the type of material behaviour when the force applied to the body decreases the body's deformation increases. Some geomaterials, for instance, rocks, demonstrate behaviour of this type at certain loads: during the compression tests the loading curves exhibit descending branch (post-peak softening). One of the possible mechanisms of the negative stiffness appearance in geomaterials is rotation of non-spherical grains. It is important to emphasize that in this case the descending branch may be reversible given that the testing machine is stiff enough (in general case it means an importance of boundary conditions). Existence of geomaterials with a negative modulus associated with rotations may have significant importance. In particular, important is understanding of the wave propagation in such materials. We study the stability of geomaterials with negative stiffness inclusions and wave propagation in it using two approaches: Cosserat continuum and discrete mass-spring models. In both cases we consider the rotational degrees of freedom in addition to the conventional translational ones. We show that despite non positiveness of the energy the materials with negative stiffness elements can be stable if certain conditions are met. In the case of Cosserat continuum the Cosserat shear modulus (the modulus relating the non-symmetrical part of shear stress and internal rotations) is allowed to assume negative values as long as its value does not exceed the value of the standard (positive) shear modulus. In the case of discrete mass-spring systems (with translational and rotational springs) the concentration of negative stiffness springs and the absolute values of negative spring stiffness are limited. The critical concentration when the system loses stability and the amplitude of the oscillations tends to infinity is equal to 1/2 and 3/5 for two- and three-dimensional cases respectively.

  16. Rats exhibit reference-dependent choice behavior.

    PubMed

    Bhatti, Mehwish; Jang, Hyeran; Kralik, Jerald D; Jeong, Jaeseung

    2014-07-01

    Human preferences depend on whether a chosen outcome appears to be a loss or a gain compared with what had been expected, i.e., in comparison to a reference point. Because reference dependence has such a strong influence on human decision-making, it is important to uncover its origins, which will in turn help delineate the underlying mechanisms. It remains unknown whether rats use reference points in decision-making, and yet, the study of rats could help address the question of whether reference dependence is evolutionarily conserved among mammals and could provide a nonhuman animal model to investigate the neural mechanisms underlying this important cognitive process. The aim of the current study was to determine whether rats show reference-dependent choice behavior. We developed a novel paradigm by modifying the "T" maze by installing "pockets" to the left and right of the "T" stem that held reward pellets so rats would potentially develop reference values for each option prior to choice. We found that the rats were indeed sensitive to the way alternatives were presented. That is, they exhibited reference-dependent choice behavior by avoiding the choice option framed as a loss (e.g., having four reward pellets in the pocket, but receiving only one), at least under conditions with certain outcomes and clear differences between the reference and outcome quantities. Despite the small number of rats in this study, this species-level capacity suggests that reference dependence in general and loss aversion in particular may be conserved traits that evolved at or before the emergence of mammals.

  17. 32 CFR 705.26 - Exhibit availability report.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... of all exhibits is required. (b) A current inventory of exhibits headquartered in Washington, DC, and...: Officer-in-Charge, Navy Recruiting Exhibit Center, Washington Navy Yard, Washington, DC 20374....

  18. 32 CFR 705.26 - Exhibit availability report.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... of all exhibits is required. (b) A current inventory of exhibits headquartered in Washington, DC, and...: Officer-in-Charge, Navy Recruiting Exhibit Center, Washington Navy Yard, Washington, DC 20374....

  19. 32 CFR 705.26 - Exhibit availability report.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... of all exhibits is required. (b) A current inventory of exhibits headquartered in Washington, DC, and...: Officer-in-Charge, Navy Recruiting Exhibit Center, Washington Navy Yard, Washington, DC 20374....

  20. 29 CFR 2204.202 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 9 2010-07-01 2010-07-01 false Net worth exhibit. 2204.202 Section 2204.202 Labor... COMMISSION Information Required From Applicants § 2204.202 Net worth exhibit. (a) Each applicant except a... exhibit showing the net worth of the applicant as of the date specified by § 2204.105(c). The exhibit...

  1. 37 CFR 1.95 - Copies of exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... COMMERCE GENERAL RULES OF PRACTICE IN PATENT CASES National Processing Provisions Models, Exhibits, Specimens § 1.95 Copies of exhibits. Copies of models or other physical exhibits will not ordinarily be furnished by the Office, and any model or exhibit in an application or patent shall not be taken from...

  2. Temporary and Travelling Exhibitions. Museums and Monuments, X.

    ERIC Educational Resources Information Center

    Daifuku, Hiroshi; And Others

    The permanent exhibition, the most typical form of museum exhibition, has failed to attract repeated visitation, since visitors quickly become familiar with the objects shown. The temporary exhibition evolved as a result for the need of repeated visitation. The temporary exhibition, set up for a period of one to six months, introduces fresh…

  3. Development of Exhibit on Arctic Climate Change Called The Arctic: A Friend Acting Strangely Exhibition

    SciTech Connect

    Stauffer, Barbara W.

    2006-04-01

    The exhibition, The Arctic: A Friend Acting Strangely, was developed at the Smithsonian Institution’s National Museum of Natural History (NMNH) as a part of the museum’s Forces of Change exhibit series on global change. It opened to the public in Spring 2006, in conjunction with another Forces of Change exhibit on the Earth’s atmosphere called Change Is in the Air. The exhibit was a 2000 square-foot presentation that explored the forces and consequences of the changing Arctic as documented by scientists and native residents alike. Native peoples of the Arctic have always lived with year-to-year fluctuations in weather and ice conditions. In recent decades, they have witnessed that the climate has become unpredictable, the land and sea unfamiliar. An elder in Arctic Canada recently described the weather as uggianaqtuq —an Inuit word that can suggest strange, unexpected behavior, sometimes described as that of “a friend acting strangely.” Scientists too have been documenting dramatic changes in the Arctic. Air temperatures have warmed over most—though not all—of the Arctic since the 1950s; Arctic precipitation may have increased by as much as 8%; seasonal melting of the Greenland Ice Sheet has increased on average by 16% since 1979; polar-orbiting satellites have measured a 15¬–20% decline in sea ice extent since the 1970s; aircraft reconnaissance and ship observations show a steady decrease in sea ice since the 1950s. In response to this warming, plant distributions have begun to shift and animals are changing their migration routes. Some of these changes may have beneficial effects while others may bring hardship or have costly implications. And, many scientists consider arctic change to be a ‘bell-weather’ for large-scale changes in other regions of the world. The exhibition included text, photos artifacts, hands-on interactives and other exhibitry that illustrated the changes being documented by indigenous people and scientists alike.

  4. 48 CFR 204.7105 - Contract exhibits and attachments.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... attachments. 204.7105 Section 204.7105 Federal Acquisition Regulations System DEFENSE ACQUISITION REGULATIONS... 204.7105 Contract exhibits and attachments. Follow the procedures at PGI 204.7105 for use and numbering of contract exhibits and attachments....

  5. 32 CFR 705.25 - Navy Exhibit Center.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... AND OFFICIAL RECORDS PUBLIC AFFAIRS REGULATIONS § 705.25 Navy Exhibit Center. (a) The center is a... mission is to produce, transport and display U.S. Navy exhibits throughout the United States. It...

  6. Swimming behavior of selected species of Archaea.

    PubMed

    Herzog, Bastian; Wirth, Reinhard

    2012-03-01

    The swimming behavior of Bacteria has been studied extensively, at least for some species like Escherichia coli. In contrast, almost no data have been published for Archaea on this topic. In a systematic study we asked how the archaeal model organisms Halobacterium salinarum, Methanococcus voltae, Methanococcus maripaludis, Methanocaldococcus jannaschii, Methanocaldococcus villosus, Pyrococcus furiosus, and Sulfolobus acidocaldarius swim and which swimming behavior they exhibit. The two Euryarchaeota M. jannaschii and M. villosus were found to be, by far, the fastest organisms reported up to now, if speed is measured in bodies per second (bps). Their swimming speeds, at close to 400 and 500 bps, are much higher than the speed of the bacterium E. coli or of a very fast animal, like the cheetah, each with a speed of ca. 20 bps. In addition, we observed that two different swimming modes are used by some Archaea. They either swim very rapidly, in a more or less straight line, or they exhibit a slower kind of zigzag swimming behavior if cells are in close proximity to the surface of the glass capillary used for observation. We argue that such a "relocate-and-seek" behavior enables the organisms to stay in their natural habitat.

  7. Weight lifting can facilitate appreciative comprehension for museum exhibits

    PubMed Central

    Yamada, Yuki; Harada, Shinya; Choi, Wonje; Fujino, Rika; Tokunaga, Akinobu; Gao, YueYun; Miura, Kayo

    2014-01-01

    Appreciation of exhibits in a museum can be equated to a virtual experience of lives in the contexts originally surrounding the exhibits. Here we focus on the importance of weight information, and hence tested whether experiencing a weight during museum exhibit appreciation affects the beholders' satisfaction and recognition memory for the exhibits. An experiment was performed at a museum exhibiting skeletal preparations of animals. We used nine preparations and prepared four weight stimuli as weight cues in accordance with the actual weight of four of the preparations: Remaining five preparations was displayed without weight stimuli. In the cued condition, participants were asked to lift up the weight stimuli during their observation of the four exhibits. In the uncued condition, participants observed the exhibits without touching the weight stimuli. After observation of the exhibits, the participants responded to a questionnaire that measured their impressions of the exhibits and the museum, and performed a recognition test on the exhibits. Results showed that memory performance was better and viewing duration was longer with weight lifting instruction than without instruction. A factor analysis on the questionnaires revealed four factors (likeability, contentment, value, and quality). A path analysis showed indirect effects of viewing duration on memory performance and willingness-to-pay (WTP) for the museum appreciation through the impression factors. Our findings provide insight into a new interactive exhibition that enables long appreciation producing positive effects on visitors' impression, memory, and value estimation for exhibits. PMID:24782807

  8. 45 CFR 1160.5 - Eligibility for domestic exhibitions.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... AND ARTIFACTS INDEMNITY ACT § 1160.5 Eligibility for domestic exhibitions. An indemnity agreement for...-owned objects; (B) Exhibitions outside of the United States of domestic-owned objects; or (C) Exhibitions in the United States of both foreign- and domestic-owned objects, with the foreign-owned...

  9. 77 FR 31420 - Culturally Significant Objects Imported for Exhibition

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-25

    ... Culturally Significant Objects Imported for Exhibition Determinations: ``Alighiero Boetti: Game Plan... determine that the objects to be included in the exhibition ``Alighiero Boetti: Game Plan'' imported from abroad for temporary exhibition within the United States, are of cultural significance. The objects...

  10. 14 CFR Appendix to Subpart A of... - Listing of Exhibits

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 5 2011-01-01 2010-01-01 true Listing of Exhibits Appendix to Subpart A of Part 1260 Aeronautics and Space NATIONAL AERONAUTICS AND SPACE ADMINISTRATION GRANTS AND COOPERATIVE AGREEMENTS General Pt. 1260, Subpt. A, App. Appendix to Subpart A of Part 1260—Listing of Exhibits Exhibit...

  11. A Major Children's Educational Art Exhibit: An Evaluative Case Study.

    ERIC Educational Resources Information Center

    Schlenk, George W.; Shrock, Sharon A.

    Results of a case study of an exhibit of art and artifacts designed for children are presented. The focus of the study was to apply the principles of instructional-message design to the evaluation of the exhibit. The exhibit, "Art Inside Out: Exploring Art and Culture through Time," was displayed at the Art Institute of Chicago. Textual elements,…

  12. 17 CFR 201.42 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... that exhibit in accordance with 17 CFR 201.190. ... 17 Commodity and Securities Exchanges 2 2010-04-01 2010-04-01 false Net worth exhibit. 201.42... Regulations Pertaining to the Equal Access to Justice Act § 201.42 Net worth exhibit. (a) Each...

  13. 39 CFR 960.10 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 39 Postal Service 1 2010-07-01 2010-07-01 false Net worth exhibit. 960.10 Section 960.10 Postal... JUSTICE ACT IN POSTAL SERVICE PROCEEDINGS Information Required From Applicants § 960.10 Net worth exhibit... with its application a detailed exhibit showing the net worth of the applicant and any affiliates...

  14. 10 CFR 1023.311 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 10 Energy 4 2010-01-01 2010-01-01 false Net worth exhibit. 1023.311 Section 1023.311 Energy... Access to Justice Act Information Required from Applicants § 1023.311 Net worth exhibit. (a) Each... application a detailed exhibit showing the net worth of the applicant and any affiliates (as defined in §...

  15. 45 CFR 13.11 - Net worth exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 45 Public Welfare 1 2010-10-01 2010-10-01 false Net worth exhibits. 13.11 Section 13.11 Public... TO JUSTICE ACT IN AGENCY PROCEEDINGS Information Required from Applicants § 13.11 Net worth exhibits. (a) Each applicant must provide with its application a detailed exhibit showing the net worth of...

  16. 49 CFR 826.22 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 7 2010-10-01 2010-10-01 false Net worth exhibit. 826.22 Section 826.22... Net worth exhibit. (a) Each applicant except a qualified tax-exempt organization or cooperative association must provide with its application a detailed exhibit showing the net worth of the applicant...

  17. 7 CFR 1.191 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 1 2010-01-01 2010-01-01 false Net worth exhibit. 1.191 Section 1.191 Agriculture... § 1.191 Net worth exhibit. (a) An applicant, except a qualified tax-exempt organization or cooperative association, must provide with its application a detailed exhibit showing the net worth of the applicant...

  18. 17 CFR 148.12 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 17 Commodity and Securities Exchanges 1 2010-04-01 2010-04-01 false Net worth exhibit. 148.12... Required from Applicants § 148.12 Net worth exhibit. (a) Each applicant except a qualified tax-exempt organization or cooperative association must provide with its application a detailed exhibit showing the...

  19. 31 CFR 6.9 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 31 Money and Finance: Treasury 1 2010-07-01 2010-07-01 false Net worth exhibit. 6.9 Section 6.9... EQUAL ACCESS TO JUSTICE ACT Information Required From Applicants § 6.9 Net worth exhibit. (a) Each... application a detailed exhibit showing the net worth of the applicant and any affiliates (as defined in §...

  20. 5 CFR 2610.202 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 3 2010-01-01 2010-01-01 false Net worth exhibit. 2610.202 Section 2610... THE EQUAL ACCESS TO JUSTICE ACT Information Required From Applicants § 2610.202 Net worth exhibit. (a... with its application a detailed exhibit showing the net worth of the applicant and any affiliates...

  1. 49 CFR 6.19 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 1 2010-10-01 2010-10-01 false Net worth exhibit. 6.19 Section 6.19... PROCEEDINGS Information Required from Applicants § 6.19 Net worth exhibit. (a) Each applicant except a... exhibit showing the net worth of the applicant and any affiliates (as defined in this part) when...

  2. 49 CFR 1016.202 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 8 2010-10-01 2010-10-01 false Net worth exhibit. 1016.202 Section 1016.202... BY PARTIES TO BOARD ADJUDICATORY PROCEEDINGS Information Required From Applicants § 1016.202 Net worth exhibit. (a) Each applicant must provide with its application a detailed exhibit showing the...

  3. African Past: Migrant Present. A Guide to the Exhibition.

    ERIC Educational Resources Information Center

    Twining, Mary Arnold; Roark-Calnek, Sue

    This exhibit guide describes an exhibition of African folk arts produced by seasonal migrant farmworkers in western New York State. Workers come from the American South, Haiti, Puerto Rico, and Jamaica. The exhibition pieces were collected through the BOCES Geneseo Migrant Center's Folk Arts Program and Creative Artists Migrant Program Services…

  4. Weight lifting can facilitate appreciative comprehension for museum exhibits.

    PubMed

    Yamada, Yuki; Harada, Shinya; Choi, Wonje; Fujino, Rika; Tokunaga, Akinobu; Gao, Yueyun; Miura, Kayo

    2014-01-01

    Appreciation of exhibits in a museum can be equated to a virtual experience of lives in the contexts originally surrounding the exhibits. Here we focus on the importance of weight information, and hence tested whether experiencing a weight during museum exhibit appreciation affects the beholders' satisfaction and recognition memory for the exhibits. An experiment was performed at a museum exhibiting skeletal preparations of animals. We used nine preparations and prepared four weight stimuli as weight cues in accordance with the actual weight of four of the preparations: Remaining five preparations was displayed without weight stimuli. In the cued condition, participants were asked to lift up the weight stimuli during their observation of the four exhibits. In the uncued condition, participants observed the exhibits without touching the weight stimuli. After observation of the exhibits, the participants responded to a questionnaire that measured their impressions of the exhibits and the museum, and performed a recognition test on the exhibits. Results showed that memory performance was better and viewing duration was longer with weight lifting instruction than without instruction. A factor analysis on the questionnaires revealed four factors (likeability, contentment, value, and quality). A path analysis showed indirect effects of viewing duration on memory performance and willingness-to-pay (WTP) for the museum appreciation through the impression factors. Our findings provide insight into a new interactive exhibition that enables long appreciation producing positive effects on visitors' impression, memory, and value estimation for exhibits.

  5. Online Cultural Heritage Exhibitions: A Survey of Strategic Issues

    ERIC Educational Resources Information Center

    Liew, Chern Li

    2006-01-01

    Purpose: This paper seeks to report findings from a study that looked at a range of strategic issues faced in the development, management and maintenance of online cultural heritage exhibitions. The study examined exhibitions from different types of cultural agencies and asked questions about whether, for instance, the exhibitions are part of the…

  6. 47 CFR 1.356 - Copies of exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 47 Telecommunication 1 2010-10-01 2010-10-01 false Copies of exhibits. 1.356 Section 1.356....356 Copies of exhibits. No document or exhibit, or part thereof, shall be received as, or admitted in... offered in evidence, copies shall be furnished to other counsel unless the presiding officer...

  7. [Energy education exhibits for Insights El Paso Science Museum

    SciTech Connect

    Shubinski, R.

    1998-05-27

    The grant in question, DE-FG03-94ER75954, was awarded to Insights El Paso Science Museum to build key exhibits. These exhibits helped the Museum fulfill its mission to ``promote curiosity and stimulate interest by exploratory, entertaining, exciting, and participatory learning in a broad range of scientific disciplines to persons of all ages regionally and internationally.`` There are several current Board of Directors members who also were Board members during the grant period and who helped construct some of the exhibits. Through speaking with them and reviewing minutes of Board meetings during 1994, it has been determined that seven of the ten proposed exhibits were constructed, with an eighth exhibit constructed as an alternative. Photos of seven of the exhibits and preliminary sketches of some are attached. Following is a list of the constructed exhibits: Hot or Cold, Give and Take, Conduction, Convection, Sources of Energy, Wind Generator, Solar Tracker, and Perpetual Motion.

  8. Engineering Hydrogen Gas Production from Formate in a Hyperthermophile by Heterologous Production of an 18-Subunit Membrane-bound Complex*

    PubMed Central

    Lipscomb, Gina L.; Schut, Gerrit J.; Thorgersen, Michael P.; Nixon, William J.; Kelly, Robert M.; Adams, Michael W. W.

    2014-01-01

    Biohydrogen gas has enormous potential as a source of reductant for the microbial production of biofuels, but its low solubility and poor gas mass transfer rates are limiting factors. These limitations could be circumvented by engineering biofuel production in microorganisms that are also capable of generating H2 from highly soluble chemicals such as formate, which can function as an electron donor. Herein, the model hyperthermophile, Pyrococcus furiosus, which grows optimally near 100 °C by fermenting sugars to produce H2, has been engineered to also efficiently convert formate to H2. Using a bacterial artificial chromosome vector, the 16.9-kb 18-gene cluster encoding the membrane-bound, respiratory formate hydrogen lyase complex of Thermococcus onnurineus was inserted into the P. furiosus chromosome and expressed as a functional unit. This enabled P. furiosus to utilize formate as well as sugars as an H2 source and to do so at both 80° and 95 °C, near the optimum growth temperature of the donor (T. onnurineus) and engineered host (P. furiosus), respectively. This accomplishment also demonstrates the versatility of P. furiosus for metabolic engineering applications. PMID:24318960

  9. Bioenergetic and physiological studies of hyperthermophilic archaea. Final report

    SciTech Connect

    Kelly, R.M.

    1999-03-01

    This project focuses on physiological and bioenergetic characteristics of two representative hyperthermophilic archaea: Thermococcus litoralis (T{sub opt} 88 C) and Pyrococcus furiosus (T{sub opt} 98 C). Both are obligately anaerobic heterotrophs which grow in the presence or absence of reducible sulfur compounds. T. litoralis was studied in relation to information previously developed for P. furiosus: effect of sulfur reduction on bioenergetics, preferred fermentation patterns, tungsten requirement, etc. A defined medium was developed for T. litoralis consisting of amino acids, vitamins and nucleotides. This serves as the basis for continuous culture studies probing metabolic response to media changes. P. furiosus and T. litoralis have also been found to produce a polysaccharide in the presence of maltose and yeast extract. The composition and chemical structure of this polysaccharide was investigated as well as the metabolic motivation for its production. A novel and, perhaps, primitive intracellular proteolytic complex (previously designated as protease S66) in P. furiosus was isolated and the gene encoding the subunit of the complex was cloned, sequenced and the protease expressed in active form in Eschericia coli. Among other issues, the role of this complex in protein turnover and stress response was examined in the context of this organism in addition to comparing it to other complexes in eubacterial and eukaryotic cells. Biochemical characteristics of the protease have been measured in addition to examining other proteolytic species in P. furiosus.

  10. 18. INTERIOR OF CIVIL RIGHTS INSTITUTE, MILESTONE GALLERY EXHIBITION OF ...

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

    18. INTERIOR OF CIVIL RIGHTS INSTITUTE, MILESTONE GALLERY EXHIBITION OF THE SIXTEENTH STREET CHURCH, LOOKING NORTHWEST - Sixteenth Street Baptist Church, 1530 Sixth Avenue North, Birmingham, Jefferson County, AL

  11. MCM ring hexamerization is a prerequisite for DNA-binding

    SciTech Connect

    Froelich, Clifford A.; Nourse, Amanda; Enemark, Eric J.

    2015-09-13

    The hexameric Minichromosome Maintenance (MCM) protein complex forms a ring that unwinds DNA at the replication fork in eukaryotes and archaea. Our recent crystal structure of an archaeal MCM N-terminal domain bound to single-stranded DNA (ssDNA) revealed ssDNA associating across tight subunit interfaces but not at the loose interfaces, indicating that DNA-binding is governed not only by the DNA-binding residues of the subunits (MCM ssDNA-binding motif, MSSB) but also by the relative orientation of the subunits. We now extend these findings to show that DNA-binding by the MCM N-terminal domain of the archaeal organism Pyrococcus furiosus occurs specifically in the hexameric oligomeric form. We show that mutants defective for hexamerization are defective in binding ssDNA despite retaining all the residues observed to interact with ssDNA in the crystal structure. One mutation that exhibits severely defective hexamerization and ssDNA-binding is at a conserved phenylalanine that aligns with the mouse Mcm4(Chaos3) mutation associated with chromosomal instability, cancer, and decreased intersubunit association.

  12. Crystal structure of the sugar binding domain of the archaeal transcriptional regulator TrmB.

    PubMed

    Krug, Michael; Lee, Sung-Jae; Diederichs, Kay; Boos, Winfried; Welte, Wolfram

    2006-04-21

    TrmB is an alpha-glucoside-sensing transcriptional regulator controlling two operons encoding maltose/trehalose and maltodextrin ABC transporters of Pyrococcus furiosus. The crystal structure of an N-terminal truncated derivative of TrmB (amino acids 2-109 deleted; TrmB(delta2-109)) was solved at 1.5 A resolution. This protein has lost its DNA binding domain but has retained its sugar recognition site. The structure represents a novel sugar-binding fold. TrmB(delta2-109) bound maltose, glucose, sucrose, and maltotriose, exhibiting Kd values of 6.8, 25, 34, and 160 microM, respectively. TrmB(delta2-109) behaved as a monomer in dilute buffer solution in contrast to the full-length protein, which is a dimer. Co-crystallization with bound maltose identified a binding site involving seven amino acid residues: Ser229, Asn305, Gly320, Met321, Val324, Ile325, and Glu326. Six of these residues interact with the nonreducing glucosyl residue of maltose. The nonreducing glucosyl residue is shared by all substrates bound to TrmB, suggesting it as a common recognition motif.

  13. Formation of the conserved pseudouridine at position 55 in archaeal tRNA.

    PubMed

    Roovers, Martine; Hale, Caryn; Tricot, Catherine; Terns, Michael P; Terns, Rebecca M; Grosjean, Henri; Droogmans, Louis

    2006-01-01

    Pseudouridine (Psi) located at position 55 in tRNA is a nearly universally conserved RNA modification found in all three domains of life. This modification is catalyzed by TruB in bacteria and by Pus4 in eukaryotes, but so far the Psi55 synthase has not been identified in archaea. In this work, we report the ability of two distinct pseudouridine synthases from the hyperthermophilic archaeon Pyrococcus furiosus to specifically modify U55 in tRNA in vitro. These enzymes are (pfu)Cbf5, a protein known to play a role in RNA-guided modification of rRNA, and (pfu)PsuX, a previously uncharacterized enzyme that is not a member of the TruB/Pus4/Cbf5 family of pseudouridine synthases. (pfu)PsuX is hereafter renamed (pfu)Pus10. Both enzymes specifically modify tRNA U55 in vitro but exhibit differences in substrate recognition. In addition, we find that in a heterologous in vivo system, (pfu)Pus10 efficiently complements an Escherichia coli strain deficient in the bacterial Psi55 synthase TruB. These results indicate that it is probable that (pfu)Cbf5 or (pfu)Pus10 (or both) is responsible for the introduction of pseudouridine at U55 in tRNAs in archaea. While we cannot unequivocally assign the function from our results, both possibilities represent unexpected functions of these proteins as discussed herein.

  14. Development of Novel Sugar Isomerases by Optimization of Active Sites in Phosphosugar Isomerases for Monosaccharides

    PubMed Central

    Yeom, Soo-Jin; Kim, Yeong-Su

    2013-01-01

    Phosphosugar isomerases can catalyze the isomerization of not only phosphosugar but also of monosaccharides, suggesting that the phosphosugar isomerases can be used as sugar isomerases that do not exist in nature. Determination of active-site residues of phosphosugar isomerases, including ribose-5-phosphate isomerase from Clostridium difficile (CDRPI), mannose-6-phosphate isomerase from Bacillus subtilis (BSMPI), and glucose-6-phosphate isomerase from Pyrococcus furiosus (PFGPI), was accomplished by docking of monosaccharides onto the structure models of the isomerases. The determinant residues, including Arg133 of CDRPI, Arg192 of BSMPI, and Thr85 of PFGPI, were subjected to alanine substitutions and found to act as phosphate-binding sites. R133D of CDRPI, R192 of BSMPI, and T85Q of PFGPI displayed the highest catalytic efficiencies for monosaccharides at each position. These residues exhibited 1.8-, 3.5-, and 4.9-fold higher catalytic efficiencies, respectively, for the monosaccharides than the wild-type enzyme. However, the activities of these 3 variant enzymes for phosphosugars as the original substrates disappeared. Thus, R133D of CDRPI, R192 of BSMPI, and T85Q of PFGPI are no longer phosphosugar isomerases; instead, they are changed to a d-ribose isomerase, an l-ribose isomerase, and an l-talose isomerase, respectively. In this study, we used substrate-tailored optimization to develop novel sugar isomerases which are not found in nature based on phosphosugar isomerases. PMID:23204422

  15. MCM ring hexamerization is a prerequisite for DNA-binding

    DOE PAGESBeta

    Froelich, Clifford A.; Nourse, Amanda; Enemark, Eric J.

    2015-09-13

    The hexameric Minichromosome Maintenance (MCM) protein complex forms a ring that unwinds DNA at the replication fork in eukaryotes and archaea. Our recent crystal structure of an archaeal MCM N-terminal domain bound to single-stranded DNA (ssDNA) revealed ssDNA associating across tight subunit interfaces but not at the loose interfaces, indicating that DNA-binding is governed not only by the DNA-binding residues of the subunits (MCM ssDNA-binding motif, MSSB) but also by the relative orientation of the subunits. We now extend these findings to show that DNA-binding by the MCM N-terminal domain of the archaeal organism Pyrococcus furiosus occurs specifically in themore » hexameric oligomeric form. We show that mutants defective for hexamerization are defective in binding ssDNA despite retaining all the residues observed to interact with ssDNA in the crystal structure. One mutation that exhibits severely defective hexamerization and ssDNA-binding is at a conserved phenylalanine that aligns with the mouse Mcm4(Chaos3) mutation associated with chromosomal instability, cancer, and decreased intersubunit association.« less

  16. 45 CFR 1160.4 - Eligibility for international exhibitions.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... AND ARTIFACTS INDEMNITY ACT § 1160.4 Eligibility for international exhibitions. An indemnity agreement...- and domestic-owned objects in an international exhibition. The foreign-owned objects are eligible for indemnity coverage under paragraph (a) of this section, and the domestic-owned objects may be eligible...

  17. 78 FR 7849 - Culturally Significant Objects Imported for Exhibition

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-02-04

    ... Culturally Significant Objects Imported for Exhibition Determinations: ``Edwardian Opulence: British Art at... April 15, 2003), I hereby determine that the objects to be included in the exhibition ``Edwardian... within the United States, are of cultural significance. The objects are imported pursuant to...

  18. 77 FR 31909 - Culturally Significant Objects Imported for Exhibition

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-30

    ... Culturally Significant Objects Imported for Exhibition Determinations: ``50th Anniversary Remembrance of the..., 2003), I hereby determine that the object to be included in the exhibition ``50th Anniversary... within the United States, is of cultural significance. The object is imported pursuant to a...

  19. 78 FR 50137 - Culturally Significant Objects Imported for Exhibition

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-16

    ... Culturally Significant Objects Imported for Exhibition Determinations: ``Dena'inaq' Huch'ulyeshi: The Dena..., 2003, I hereby determine that the objects to be included in the exhibition ``Dena'inaq' Huch'ulyeshi..., are of cultural significance. The objects are imported pursuant to loan agreements with the...

  20. 75 FR 6079 - Culturally Significant Objects Imported for Exhibition

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-05

    ... Culturally Significant Objects Imported for Exhibition Determinations: ``Compass and Rule: Architecture as... 15, 2003 , I hereby determine that the objects to be included in the exhibition ``Compass and Rule... within the United States, are of cultural significance. The objects are imported pursuant to...

  1. 78 FR 25337 - Culturally Significant Object Imported for Exhibition

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-30

    ... Culturally Significant Object Imported for Exhibition Determinations: ``Bronze Statue of a Boxer, Hellenistic..., 2003), I hereby determine that the object to be included in the exhibition ``Bronze Statue of a Boxer... significance. The object is imported pursuant to a loan agreement with the foreign owner or custodian. I...

  2. 77 FR 18295 - Culturally Significant Objects Imported for Exhibition

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-27

    ... Culturally Significant Objects Imported for Exhibition Determinations: ``Roy Lichtenstein: A Retrospective... determine that the objects to be included in the exhibition ``Roy Lichtenstein: A Retrospective'' imported... objects are imported pursuant to loan agreements with the foreign owners or custodians. I also...

  3. 76 FR 68808 - Culturally Significant Objects Imported for Exhibition

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-11-07

    ... Culturally Significant Objects Imported for Exhibition Determinations: ``Transition to Christianity: Art of... April 15, 2003), I hereby determine that the objects to be included in the exhibition ``Transition to... the United States, are of cultural significance. The objects are imported pursuant to loan...

  4. A Critical Appraisal of State Level Science Exhibition

    ERIC Educational Resources Information Center

    Nath, Baiju K.

    2007-01-01

    Science exhibitions are really great opportunities to students as well as teachers to disseminate knowledge that they have, and to experience a variety of new inventions and innovations that also need wide dissemination. The great significance of exhibition is that it fosters acquisition of different process skills leading to the development of…

  5. 7 CFR Exhibits A-F to Subpart A... - [Reserved

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 14 2013-01-01 2013-01-01 false A Exhibits A-F to Subpart A of Part 1955 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... Real and Chattel Property Exhibits A-F to Subpart A of Part 1955...

  6. 7 CFR Exhibit A to Subpart A of... - [Reserved

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 12 2011-01-01 2011-01-01 false A Exhibit A to Subpart A of Part 1902 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... SUPERVISED BANK ACCOUNTS Supervised Bank Accounts of Loan, Grant, and Other Funds Exhibit A to Subpart A...

  7. 7 CFR Exhibit A to Subpart A of... - [Reserved

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 12 2014-01-01 2013-01-01 true A Exhibit A to Subpart A of Part 1902 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... SUPERVISED BANK ACCOUNTS Supervised Bank Accounts of Loan, Grant, and Other Funds Exhibit A to Subpart A...

  8. 7 CFR Exhibit A to Subpart A of... - [Reserved

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 12 2012-01-01 2012-01-01 false A Exhibit A to Subpart A of Part 1902 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... SUPERVISED BANK ACCOUNTS Supervised Bank Accounts of Loan, Grant, and Other Funds Exhibit A to Subpart A...

  9. 7 CFR Exhibit A to Subpart A of... - [Reserved

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 12 2013-01-01 2013-01-01 false A Exhibit A to Subpart A of Part 1902 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... SUPERVISED BANK ACCOUNTS Supervised Bank Accounts of Loan, Grant, and Other Funds Exhibit A to Subpart A...

  10. 7 CFR Exhibits A-F to Subpart A... - [Reserved

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 14 2014-01-01 2014-01-01 false A Exhibits A-F to Subpart A of Part 1955 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... Real and Chattel Property Exhibits A-F to Subpart A of Part 1955...

  11. 7 CFR Exhibits A-F to Subpart A... - [Reserved

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 14 2011-01-01 2011-01-01 false A Exhibits A-F to Subpart A of Part 1955 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... Real and Chattel Property Exhibits A-F to Subpart A of Part 1955...

  12. 7 CFR Exhibits A-F to Subpart A... - [Reserved

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 14 2012-01-01 2012-01-01 false A Exhibits A-F to Subpart A of Part 1955 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... Real and Chattel Property Exhibits A-F to Subpart A of Part 1955...

  13. Designing Art Exhibitions in an Educational Virtual World

    ERIC Educational Resources Information Center

    Julian, June; Crooks, Julian

    2011-01-01

    Demonstrating the multiple features of the Cerulean Gallery in Second Life, this research report showcases several exemplar exhibits created by students, artists, and museums. Located in The Educational Media Center, a Second Life teaching and social space, the Cerulean Gallery exhibits functioned as case studies that tested its effectiveness as…

  14. A Commemorative History of the George Rogers Clark Bicentennial Exhibit.

    ERIC Educational Resources Information Center

    Indiana State Museum Society, Inc., Indianapolis.

    This pamphlet provides an illustrated narrative history of the George Rogers Clark Bicentennial Exhibit at the Indiana State Museum. George Rogers Clark was a frontier hero of the American Revolution who explored and conquered territory in Kentucky, Ohio, and Illinois. The multimedia exhibit is open to the public from February 25, 1976 through…

  15. Making Your Trade Fair Exhibit More Productive and More Interesting.

    ERIC Educational Resources Information Center

    Newcomb, Jack

    1982-01-01

    Suggestions for producing a successful exhibit booth include the following: the effectiveness of an exhibit depends on the effectiveness of the people staffing it; avoid games and unrelated giveaway items; demonstrate product in the booth; give special attention to existing customers; and make literature available only from the booth personnel.…

  16. Using Museum Exhibits: An Innovation in Experiential Learning

    ERIC Educational Resources Information Center

    Das, Satarupa

    2015-01-01

    Museum exhibits can be a tool in experiential learning. While instructors have documented various methods of experiential learning, they have not sufficiently explored such learning from museum exhibits. Museum researchers, however, have long found a satisfying cognitive component to museum visits. This paper narrates the author's design to…

  17. 40 CFR 17.12 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 1 2010-07-01 2010-07-01 false Net worth exhibit. 17.12 Section 17.12... JUSTICE ACT IN EPA ADMINISTRATIVE PROCEEDINGS Information Required From Applicants § 17.12 Net worth... submit with its application a detailed exhibit showing its net worth at the time the proceeding...

  18. 10 CFR 12.202 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ..., 10 CFR part 9, subpart A. ... 10 Energy 1 2010-01-01 2010-01-01 false Net worth exhibit. 12.202 Section 12.202 Energy NUCLEAR... Required From Applicants § 12.202 Net worth exhibit. (a) Each applicant, except a qualified...

  19. 28 CFR 24.202 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 28 Judicial Administration 1 2010-07-01 2010-07-01 false Net worth exhibit. 24.202 Section 24.202... DEPARTMENT OF JUSTICE ADMINISTRATIVE PROCEEDINGS Information Required From Applicants § 24.202 Net worth... submit with its application a detailed exhibit showing its net worth at the time the proceeding...

  20. 14 CFR 1262.202 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... disposed of in accordance with the Agency's regulations under the Freedom of Information Act, at 14 CFR... 14 Aeronautics and Space 5 2010-01-01 2010-01-01 false Net worth exhibit. 1262.202 Section 1262... AGENCY PROCEEDINGS Information Required From Applicants § 1262.202 Net worth exhibit. (a) Each...

  1. 15 CFR 18.12 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... accordance with the Department's established procedures under the Freedom of Information Act (15 CFR Part 4). ... 15 Commerce and Foreign Trade 1 2010-01-01 2010-01-01 false Net worth exhibit. 18.12 Section 18.12... Information Required from Applicants § 18.12 Net worth exhibit. (a) Each applicant except a qualified...

  2. 47 CFR 1.1512 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 47 Telecommunication 1 2010-10-01 2010-10-01 false Net worth exhibit. 1.1512 Section 1.1512... Access to Justice Act (EAJA) in Agency Proceedings Information Required from Applicants § 1.1512 Net... must provide with its application a detailed exhibit showing the net worth of the applicant and...

  3. 22 CFR 134.12 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 22 Foreign Relations 1 2010-04-01 2010-04-01 false Net worth exhibit. 134.12 Section 134.12... Information Required From Applicants § 134.12 Net worth exhibit. (a) Each applicant except a qualifed tax... showing the net worth of the applicant and any affiliates (as defined in § 960.4(f)) when the...

  4. 29 CFR 16.202 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 1 2010-07-01 2010-07-01 true Net worth exhibit. 16.202 Section 16.202 Labor Office of the Secretary of Labor EQUAL ACCESS TO JUSTICE ACT Information Required From Applicants § 16.202 Net worth... provide with its application a detailed exhibit showing the net worth of the applicant and any...

  5. 14 CFR 14.11 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Net worth exhibit. 14.11 Section 14.11... IMPLEMENTING THE EQUAL ACCESS TO JUSTICE ACT OF 1980 Information Required From Applicants § 14.11 Net worth... provide with its application a detailed exhibit showing the net worth of the applicant and any...

  6. 14 CFR 77.59 - Subpoenas of witnesses and exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 2 2010-01-01 2010-01-01 false Subpoenas of witnesses and exhibits. 77.59 Section 77.59 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED) AIRSPACE OBJECTS AFFECTING NAVIGABLE AIRSPACE Rules of Practice for Hearings Under Subpart D § 77.59 Subpoenas of witnesses and exhibits....

  7. 7 CFR Exhibits A-F to Subpart A... - [Reserved

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 14 2010-01-01 2009-01-01 true A Exhibits A-F to Subpart A of Part 1955 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... Real and Chattel Property Exhibits A-F to Subpart A of Part 1955...

  8. 7 CFR Exhibit A to Subpart A of... - [Reserved

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 12 2010-01-01 2010-01-01 false A Exhibit A to Subpart A of Part 1902 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... SUPERVISED BANK ACCOUNTS Supervised Bank Accounts of Loan, Grant, and Other Funds Exhibit A to Subpart A...

  9. Cooperative Exhibition: A New Concept for the Apparel Design Teacher.

    ERIC Educational Resources Information Center

    Furer, Gloria

    1982-01-01

    Explains how an apparel and textile exhibition can benefit the institution that underwrites the project, the department that sponsors it, the community, and the apparel design teacher (through the chance for professional recognition). Methods for planning such an exhibition are given. (CT)

  10. 7 CFR Exhibit C to Subpart N of... - [Reserved

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 13 2012-01-01 2012-01-01 false C Exhibit C to Subpart N of Part 1944 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... REGULATIONS (CONTINUED) HOUSING Housing Preservation Grants Exhibit C to Subpart N of Part 1944...

  11. Learning Times Two: Creating Learning through a Children's Museum Exhibit

    ERIC Educational Resources Information Center

    Sharon, Tanya

    2012-01-01

    This article describes an innovative lab course in which students designed and constructed five developmentally (and locally) appropriate museum exhibits around the theme "Healthy Living." They installed these at a local children's museum, designed a pretest/posttest to assess children's learning from the exhibit, conducted children through the…

  12. 7 CFR Exhibit A to Subpart L of... - [Reserved

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 13 2010-01-01 2009-01-01 true A Exhibit A to Subpart L of Part 1940 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... Exhibit A to Subpart L of Part 1940...

  13. An Exhibition on Everyday Chemistry. Communicating Chemistry to the Public.

    ERIC Educational Resources Information Center

    Ucko, David A.; And Others

    1986-01-01

    Discusses a recent addition to the Museum of Science and Industry (Chicago) known as "Everyday Chemistry." This permanent exhibit on modern chemistry incorporates demonstrations of chemical reactions in ways intended to enhance public understanding. Describes the six cases in the exhibit and the automated aspects of their demonstrations. (TW)

  14. 46 CFR 169.805 - Exhibition of merchant mariner credentials.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 7 2013-10-01 2013-10-01 false Exhibition of merchant mariner credentials. 169.805 Section 169.805 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) NAUTICAL SCHOOLS SAILING SCHOOL VESSELS Operations § 169.805 Exhibition of merchant mariner credentials. Officers on any...

  15. 46 CFR 169.805 - Exhibition of merchant mariner credentials.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 7 2010-10-01 2010-10-01 false Exhibition of merchant mariner credentials. 169.805 Section 169.805 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) NAUTICAL SCHOOLS SAILING SCHOOL VESSELS Operations § 169.805 Exhibition of merchant mariner credentials. Officers on any...

  16. 46 CFR 169.805 - Exhibition of merchant mariner credentials.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 7 2014-10-01 2014-10-01 false Exhibition of merchant mariner credentials. 169.805 Section 169.805 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) NAUTICAL SCHOOLS SAILING SCHOOL VESSELS Operations § 169.805 Exhibition of merchant mariner credentials. Officers on any...

  17. 46 CFR 169.805 - Exhibition of merchant mariner credentials.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 7 2012-10-01 2012-10-01 false Exhibition of merchant mariner credentials. 169.805 Section 169.805 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) NAUTICAL SCHOOLS SAILING SCHOOL VESSELS Operations § 169.805 Exhibition of merchant mariner credentials. Officers on any...

  18. 46 CFR 169.805 - Exhibition of merchant mariner credentials.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 7 2011-10-01 2011-10-01 false Exhibition of merchant mariner credentials. 169.805 Section 169.805 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) NAUTICAL SCHOOLS SAILING SCHOOL VESSELS Operations § 169.805 Exhibition of merchant mariner credentials. Officers on any...

  19. Gifted Kids Create a Large Scale Museum Exhibit in Indianapolis.

    ERIC Educational Resources Information Center

    Waters, Mary Lou; Bostwick, Alice

    1989-01-01

    Indianapolis gifted students in grades five-seven created a museum exhibit to express the views of youth while giving adults an opportunity to listen. The process involved brainstorming, selecting a topic (drug and alcohol abuse), gathering information, forming committees, building a scale model, and creating the actual exhibit. (JDD)

  20. Exhibitions: Connecting Classroom Assessment with Culminating Demonstrations of Mastery

    ERIC Educational Resources Information Center

    Davidson, Jill

    2009-01-01

    Exhibitions are public demonstrations of mastery that occur at culminating moments, such as at the conclusion of a unit of study, the transition from one level of schooling to the next, and graduation. Exhibitions require students to speak publicly, use evidence, present engaging visual displays, and otherwise demonstrate mastery to educators,…

  1. 32 CFR 705.25 - Navy Exhibit Center.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 32 National Defense 5 2014-07-01 2014-07-01 false Navy Exhibit Center. 705.25 Section 705.25 National Defense Department of Defense (Continued) DEPARTMENT OF THE NAVY UNITED STATES NAVY REGULATIONS AND OFFICIAL RECORDS PUBLIC AFFAIRS REGULATIONS § 705.25 Navy Exhibit Center. (a) The center is...

  2. 32 CFR 705.25 - Navy Exhibit Center.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 5 2010-07-01 2010-07-01 false Navy Exhibit Center. 705.25 Section 705.25 National Defense Department of Defense (Continued) DEPARTMENT OF THE NAVY UNITED STATES NAVY REGULATIONS AND OFFICIAL RECORDS PUBLIC AFFAIRS REGULATIONS § 705.25 Navy Exhibit Center. (a) The center is...

  3. 32 CFR 705.25 - Navy Exhibit Center.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 32 National Defense 5 2012-07-01 2012-07-01 false Navy Exhibit Center. 705.25 Section 705.25 National Defense Department of Defense (Continued) DEPARTMENT OF THE NAVY UNITED STATES NAVY REGULATIONS AND OFFICIAL RECORDS PUBLIC AFFAIRS REGULATIONS § 705.25 Navy Exhibit Center. (a) The center is...

  4. 32 CFR 705.25 - Navy Exhibit Center.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 32 National Defense 5 2013-07-01 2013-07-01 false Navy Exhibit Center. 705.25 Section 705.25 National Defense Department of Defense (Continued) DEPARTMENT OF THE NAVY UNITED STATES NAVY REGULATIONS AND OFFICIAL RECORDS PUBLIC AFFAIRS REGULATIONS § 705.25 Navy Exhibit Center. (a) The center is...

  5. Modelling the Future: Exhibitions and the Materiality of Education

    ERIC Educational Resources Information Center

    Lawn, Martin, Ed.

    2009-01-01

    The role of World Exhibitions in the 19th and early 20th centuries was to confirm a relation between the nation state and modernity. As a display about industries, inventions and identities, the Exhibition, in a sense, put entire nations into an elevated, viewable space. It is a significant element in modernity as comparisons can be made, progress…

  6. 7 CFR Exhibit G to Subpart G of... - [Reserved

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 13 2010-01-01 2009-01-01 true G Exhibit G to Subpart G of Part 1940 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... REGULATIONS (CONTINUED) GENERAL Environmental Program Exhibit G to Subpart G of Part 1940...

  7. 7 CFR Exhibit G to Subpart A of... - Performance Bond

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 12 2010-01-01 2010-01-01 false Performance Bond G Exhibit G to Subpart A of Part 1924 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL.... 1924, Subpt. A, Exh. G Exhibit G to Subpart A of Part 1924—Performance Bond KNOW ALL PERSONS BY...

  8. 17 CFR 229.1016 - (Item 1016) Exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 1016) Exhibits. File as an exhibit to the schedule: (a) Any disclosure materials furnished to security...-9 Disclosure Material X X X Loan Agreement X X Report, Opinion or Appraisal X Contracts, Arrangements or Understandings X X X Statement re: Appraisal Rights X Oral Solicitation Materials X X......

  9. Perspectives on ... Multiculturalism and Library Exhibits: Sites of Contested Representation

    ERIC Educational Resources Information Center

    Reece, Gwendolyn J.

    2005-01-01

    This article analyzes a multicultural library exhibit presenting the Palestinian/Israeli conflict as a site of contested representation. Qualitative methodology is used to interrogate the exhibit and its audience reception. Drawing on insights from critical pedagogy, implications for libraries arising from this case study are given and suggestions…

  10. 7 CFR Exhibit C to Subpart N of... - [Reserved

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 13 2010-01-01 2009-01-01 true C Exhibit C to Subpart N of Part 1944 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... REGULATIONS (CONTINUED) HOUSING Housing Preservation Grants Exhibit C to Subpart N of Part 1944...

  11. 7 CFR Exhibit C to Subpart N of... - [Reserved

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 13 2011-01-01 2009-01-01 true C Exhibit C to Subpart N of Part 1944 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... REGULATIONS (CONTINUED) HOUSING Housing Preservation Grants Exhibit C to Subpart N of Part 1944...

  12. 7 CFR Exhibit F to Subpart A of... - Payment Bond

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 12 2013-01-01 2013-01-01 false Payment Bond F Exhibit F to Subpart A of Part 1924 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS.... A, Exh. F Exhibit F to Subpart A of Part 1924—Payment Bond KNOW ALL PERSONS BY THESE PRESENTS:...

  13. 7 CFR Exhibit F to Subpart A of... - [Reserved

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 14 2014-01-01 2014-01-01 false F Exhibit F to Subpart A of Part 1962 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... REGULATIONS (CONTINUED) PERSONAL PROPERTY Servicing and Liquidation of Chattel Security Exhibit F to Subpart...

  14. 7 CFR Exhibit F to Subpart A of... - [Reserved

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 14 2013-01-01 2013-01-01 false F Exhibit F to Subpart A of Part 1962 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... REGULATIONS (CONTINUED) PERSONAL PROPERTY Servicing and Liquidation of Chattel Security Exhibit F to Subpart...

  15. 7 CFR Exhibit F to Subpart A of... - [Reserved

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 14 2011-01-01 2011-01-01 false F Exhibit F to Subpart A of Part 1962 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... REGULATIONS (CONTINUED) PERSONAL PROPERTY Servicing and Liquidation of Chattel Security Exhibit F to Subpart...

  16. 7 CFR Exhibit F to Subpart A of... - Payment Bond

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 12 2014-01-01 2013-01-01 true Payment Bond F Exhibit F to Subpart A of Part 1924 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS.... A, Exh. F Exhibit F to Subpart A of Part 1924—Payment Bond KNOW ALL PERSONS BY THESE PRESENTS:...

  17. Online Cultural Heritage Exhibitions: A Survey of Information Retrieval Features

    ERIC Educational Resources Information Center

    Liew, Chern Li

    2005-01-01

    Purpose: What kinds of online cultural heritage exhibitions are now available on the internet? How far have these cultural heritage institutions voyaged in terms of harnessing the power of information and communication technology and the interactivity of multimedia systems to exhibit cultural heritage resources? This study aims to highlight the…

  18. The Development of Validated Museum Exhibits. Final Report.

    ERIC Educational Resources Information Center

    Nicol, Elizabeth H.

    Exhibit development, as conceived in this report, is an evolutionary process, drawing the museum visitor into the collaborative venture of testing and improving the exhibits. The findings of contemporary learning research were put to work in the arrangement of activities and specimens that engaged children through self-instructional sequences. The…

  19. Outreach to Science Faculty and Students through Research Exhibitions

    ERIC Educational Resources Information Center

    Chan, Tina; Hebblethwaite, Chris

    2014-01-01

    Penfield Library at the State University of New York at Oswego (SUNY Oswego) has a gallery exhibit space near the front entrance that is used to showcase student-faculty research and art class projects. This article features the library's outreach efforts to science faculty and students through research exhibitions. The library held an…

  20. 7 CFR Exhibit A to Subpart L of... - [Reserved

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 13 2012-01-01 2012-01-01 false A Exhibit A to Subpart L of Part 1940 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... Exhibit A to Subpart L of Part 1940...

  1. 7 CFR Exhibit A to Subpart L of... - [Reserved

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 13 2014-01-01 2013-01-01 true A Exhibit A to Subpart L of Part 1940 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... Exhibit A to Subpart L of Part 1940...

  2. 7 CFR Exhibit A to Subpart L of... - [Reserved

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 13 2011-01-01 2009-01-01 true A Exhibit A to Subpart L of Part 1940 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... Exhibit A to Subpart L of Part 1940...

  3. 7 CFR Exhibit A to Subpart L of... - [Reserved

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 13 2013-01-01 2013-01-01 false A Exhibit A to Subpart L of Part 1940 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... Exhibit A to Subpart L of Part 1940...

  4. 7 CFR Exhibit F to Subpart A of... - Payment Bond

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 12 2010-01-01 2010-01-01 false Payment Bond F Exhibit F to Subpart A of Part 1924 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS.... A, Exh. F Exhibit F to Subpart A of Part 1924—Payment Bond KNOW ALL PERSONS BY THESE PRESENTS:...

  5. 7 CFR Exhibit F to Subpart A of... - [Reserved

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 14 2010-01-01 2009-01-01 true F Exhibit F to Subpart A of Part 1962 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE... REGULATIONS (CONTINUED) PERSONAL PROPERTY Servicing and Liquidation of Chattel Security Exhibit F to Subpart...

  6. 46 CFR Exhibit No. 1 to Subpart F... - [Reserved

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 9 2010-10-01 2010-10-01 false No. Exhibit No. 1 to Subpart F of Part 502 Shipping FEDERAL MARITIME COMMISSION GENERAL AND ADMINISTRATIVE PROVISIONS RULES OF PRACTICE AND PROCEDURE Settlement; Prehearing Procedure Exhibit No. 1 to Subpart F of Part 502...

  7. 46 CFR Exhibit No. 1 to Subpart F... - [Reserved

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 9 2011-10-01 2011-10-01 false No. Exhibit No. 1 to Subpart F of Part 502 Shipping FEDERAL MARITIME COMMISSION GENERAL AND ADMINISTRATIVE PROVISIONS RULES OF PRACTICE AND PROCEDURE Settlement; Prehearing Procedure Exhibit No. 1 to Subpart F of Part 502...

  8. First major museum exhibit on the science of AIDS.

    PubMed

    Nary, G

    1995-05-01

    The Chicago Museum of Science and Industry recently opened its 4,000-square-foot permanent exhibit on the science of AIDS. The exhibit is designed to educate visitors about immunology and virology by presenting AIDS as an example of an immune system breakdown and showing advances made in research and prevention of diseases caused by viruses. The exhibit also seeks to engage preteens with compelling graphics and interactive displays in the hope of motivating them to pursue careers in science. One exhibit, called Frontline, allows visitors to question several physicians and researchers specializing in HIV/AIDS through a touch-screen video program. The exhibit allows these professionals to reveal their commitment and explain their views on the future of the pandemic.

  9. Exhibit Development: The Importance of Process and Evaluation

    NASA Astrophysics Data System (ADS)

    Dusenbery, P.; McLain, B.

    2010-08-01

    The Space Science Institute (SSI) is a national leader in developing national traveling exhibitions on space science education (e.g. Electric Space, MarsQuest, Alien Earths, Giant Worlds, Asteroids, and Discover Space). It is also known for developing effective digital media programs (e.g. www.alienearths.org), education workshops for formal and informal educators, and educational films (e.g. Inspire Me: Weightless Flights of Discovery). This paper focuses on the exhibit development process, spanning conceptual planning, design development, fabrication and launch. SSI's exhibit programs also include education and outreach programming and the development of an online version of the exhibit. Examples from Giant Worlds and Asteroids will be used to illustrate these development phases especially the importance of evaluation/research in exhibit development using a logic model approach.

  10. First major museum exhibit on the science of AIDS.

    PubMed

    Nary, G

    1995-05-01

    The Chicago Museum of Science and Industry recently opened its 4,000-square-foot permanent exhibit on the science of AIDS. The exhibit is designed to educate visitors about immunology and virology by presenting AIDS as an example of an immune system breakdown and showing advances made in research and prevention of diseases caused by viruses. The exhibit also seeks to engage preteens with compelling graphics and interactive displays in the hope of motivating them to pursue careers in science. One exhibit, called Frontline, allows visitors to question several physicians and researchers specializing in HIV/AIDS through a touch-screen video program. The exhibit allows these professionals to reveal their commitment and explain their views on the future of the pandemic. PMID:11362508

  11. Novel type of ADP-forming acetyl coenzyme A synthetase in hyperthermophilic archaea: heterologous expression and characterization of isoenzymes from the sulfate reducer Archaeoglobus fulgidus and the methanogen Methanococcus jannaschii.

    PubMed

    Musfeldt, Meike; Schönheit, Peter

    2002-02-01

    Acetyl coenzyme A (CoA) synthetase (ADP forming) (ACD) represents a novel enzyme of acetate formation and energy conservation (acetyl-CoA + ADP + P(i) right harpoon over left harpoon acetate + ATP + CoA) in Archaea and eukaryotic protists. The only characterized ACD in archaea, two isoenzymes from the hyperthermophile Pyrococcus furiosus, constitute 145-kDa heterotetramers (alpha(2), beta(2)). The coding genes for the alpha and beta subunits are located at different sites in the P. furiosus chromosome. Based on significant sequence similarity of the P. furiosus genes, five open reading frames (ORFs) encoding putative ACD were identified in the genome of the hyperthermophilic sulfate-reducing archaeon Archaeoglobus fulgidus and one ORF was identified in the hyperthermophilic methanogen Methanococcus jannaschii. The ORFs constitute fusions of the homologous P. furiosus genes encoding the alpha and beta subunits. Two ORFs, AF1211 and AF1938, of A. fulgidus and ORF MJ0590 of M. jannaschii were cloned and functionally overexpressed in Escherichia coli. The purified recombinant proteins were characterized as distinctive isoenzymes of ACD with different substrate specificities. In contrast to the Pyrococcus ACD, the ACDs of Archaeoglobus and Methanococcus constitute homodimers of about 140 kDa composed of two identical 70-kDa subunits, which represent fusions of the homologous P. furiosus alpha and beta subunits in an alphabeta (AF1211 and MJ0590) or betaalpha (AF1938) orientation. The data indicate that A. fulgidus and M. jannaschii contains a novel type of ADP-forming acetyl-CoA synthetase in Archaea, in which the subunit polypeptides and their coding genes are fused.

  12. Maine Exhibitions Assessment Project, September 2002-June 2004. Technical Criteria for Including Exhibition Assessments in Comprehensive Local Assessment Systems. Revised.

    ERIC Educational Resources Information Center

    Maine Department of Education, 2004

    2004-01-01

    In September 2002, the Maine Department of Education began organizing an Exhibition Assessment Advisory Committee whose purpose it was to consider meaningful ways to include exhibition assessments in comprehensive local assessment systems. School administrators were invited to participate and/or nominate practitioners in their districts to become…

  13. Statistical properties of chaotic dynamical systems which exhibit strange attractors

    SciTech Connect

    Jensen, R.V.; Oberman, C.R.

    1981-07-01

    A path integral method is developed for the calculation of the statistical properties of turbulent dynamical systems. The method is applicable to conservative systems which exhibit a transition to stochasticity as well as dissipative systems which exhibit strange attractors. A specific dissipative mapping is considered in detail which models the dynamics of a Brownian particle in a wave field with a broad frequency spectrum. Results are presented for the low order statistical moments for three turbulent regimes which exhibit strange attractors corresponding to strong, intermediate, and weak collisional damping.

  14. 27 CFR 7.42 - Exhibiting certificates to Government officials.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF MALT BEVERAGES Requirements for Approval of Labels of Malt Beverages Domestically Bottled or Packed § 7.42 Exhibiting...

  15. 27 CFR 7.42 - Exhibiting certificates to Government officials.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF MALT BEVERAGES Requirements for Approval of Labels of Malt Beverages Domestically Bottled or Packed § 7.42 Exhibiting...

  16. 27 CFR 7.42 - Exhibiting certificates to Government officials.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... TRADE BUREAU, DEPARTMENT OF THE TREASURY ALCOHOL LABELING AND ADVERTISING OF MALT BEVERAGES Requirements for Approval of Labels of Malt Beverages Domestically Bottled or Packed § 7.42 Exhibiting...

  17. 27 CFR 7.42 - Exhibiting certificates to Government officials.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF MALT BEVERAGES Requirements for Approval of Labels of Malt Beverages Domestically Bottled or Packed § 7.42 Exhibiting...

  18. 45 CFR 1160.4 - Eligibility for international exhibitions.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... AND ARTIFACTS INDEMNITY ACT § 1160.4 Eligibility for international exhibitions. An indemnity agreement... include more than 150 works of art by Impressionist painter Auguste Renoir. Museums in Paris and...

  19. 45 CFR 1160.4 - Eligibility for international exhibitions.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... AND ARTIFACTS INDEMNITY ACT § 1160.4 Eligibility for international exhibitions. An indemnity agreement... include more than 150 works of art by Impressionist painter Auguste Renoir. Museums in Paris and...

  20. 45 CFR 1160.4 - Eligibility for international exhibitions.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... AND ARTIFACTS INDEMNITY ACT § 1160.4 Eligibility for international exhibitions. An indemnity agreement... include more than 150 works of art by Impressionist painter Auguste Renoir. Museums in Paris and...

  1. 45 CFR 1160.4 - Eligibility for international exhibitions.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... AND ARTIFACTS INDEMNITY ACT § 1160.4 Eligibility for international exhibitions. An indemnity agreement... include more than 150 works of art by Impressionist painter Auguste Renoir. Museums in Paris and...

  2. 76 FR 61472 - Culturally Significant Objects Imported for Exhibition

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-04

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF STATE Culturally Significant Objects Imported for Exhibition Determinations: ``The Game of Kings: Medieval Ivory... Kings: Medieval Ivory Chessmen from the Isle of Lewis,'' imported from abroad for temporary...

  3. Unimode metamaterials exhibiting negative linear compressibility and negative thermal expansion

    NASA Astrophysics Data System (ADS)

    Dudek, Krzysztof K.; Attard, Daphne; Caruana-Gauci, Roberto; Wojciechowski, Krzysztof W.; Grima, Joseph N.

    2016-02-01

    Unimode metamaterials made from rotating rigid triangles are analysed mathematically for their mechanical and thermal expansion properties. It is shown that these unimode systems exhibit positive Poisson’s ratios irrespective of size, shape and angle of aperture, with the Poisson’s ratio exhibiting giant values for certain conformations. When the Poisson’s ratio in one loading direction is larger than +1, the systems were found to exhibit the anomalous property of negative linear compressibility along this direction, that is, the systems expand in this direction when hydrostatically compressed. Also discussed are the thermal expansion properties of these systems under the assumption that the units exhibit increased rotational agitation once subjected to an increase in temperature. The effect of the geometric parameters on the aforementioned thermo-mechanical properties of the system, are discussed, with the aim of identifying negative behaviour.

  4. Science, providence, and progress at the Great Exhibition.

    PubMed

    Cantor, Geoffrey

    2012-09-01

    The Great Exhibition of 1851 is generally interpreted as a thoroughly secular event that celebrated progress in science, technology, and industry. In contrast to this perception, however, the exhibition was viewed by many contemporaries as a religious event of considerable importance. Although some religious commentators were highly critical of the exhibition and condemned the display of artifacts in the Crystal Palace as giving succor to materialism, others incorporated science and technology into their religious frameworks. Drawing on sermons, tracts, and the religious periodical press, this essay pays close attention to the ways in which science and technology were endowed with providentialist significance and particularly examines the notion of human progress used by a number of Christian writers, especially Congregationalists, who set scientific and technological progress within a teleological religious perspective. This discussion sheds fresh light not only on the Great Exhibition itself but also on the deployment of natural theology in mid-nineteenth-century Britain.

  5. 78 FR 21979 - Federal Advisory Committee on International Exhibitions Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-12

    ... From the Federal Register Online via the Government Publishing Office NATIONAL FOUNDATION ON THE ARTS AND THE HUMANITIES Federal Advisory Committee on International Exhibitions Meeting AGENCY..., notice is hereby given that one meeting of the Federal Advisory Committee on International...

  6. 78 FR 68479 - Federal Advisory Committee on International Exhibitions Meeting

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-14

    ... From the Federal Register Online via the Government Publishing Office NATIONAL FOUNDATION ON THE ARTS AND THE HUMANITIES Federal Advisory Committee on International Exhibitions Meeting AGENCY..., notice is hereby given that one meeting of the Federal Advisory Committee on International...

  7. "Wenn die Erde bebt", an educational public exhibition in Vienna

    NASA Astrophysics Data System (ADS)

    Parithusta, Rizkita; Brueckl, Ewald; Heuer, Rudolf; Mitterbauer, Ulrike

    2010-05-01

    Natural disasters can cause the loss of human lives, an economic crisis and also the loss of irreplaceable cultural heritage. Earthquakes can mean instantaneous destruction without warning, causing extensive and often irreparable damage to our heritage. An exhibition with the title "Wenn die Erde bebt" (i.e. "When the Earth shakes") which was held at the Natural History Museum, Vienna; in an effort to introduce understanding, awareness, and preparedness to the public, facing earthquake phenomenon. The exhibition compiled and classified examples of large earthquakes and introduces into the basic principles of seismology. It further addresses earthquake impact and how to live with earthquakes, giving access to the most suitable procedure of safety education. The idea of the exhibition is communicated by the means of posters, videos, and physical models which support the understanding of seismometry, elastic rebound theory and earthquake resistant construction. The exhibition is an Austrian contribution to IYPE - International Year of Planet Earth and is now on tour through Austria.

  8. 17. INTERIOR VIEW OF BASEMENT EXHIBITION OF EVENTS OF CIVIL ...

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

    17. INTERIOR VIEW OF BASEMENT EXHIBITION OF EVENTS OF CIVIL RIGHTS MOVEMENT AND THE 1963 BOMBING OF THE CHURCH, LOOKING SOUTH - Sixteenth Street Baptist Church, 1530 Sixth Avenue North, Birmingham, Jefferson County, AL

  9. JoAnn Morgan looks at Newscapade exhibit.

    NASA Technical Reports Server (NTRS)

    2000-01-01

    JoAnn Morgan, associate director for Advanced Development and Shuttle Upgrades at KSC, studies posters of space-related news stories in the mobile exhibition called 'NewsCapade with Al Neuharth.' The exhibit started its cross-country tour in San Francisco in April. It is a traveling version of the Newseum in Arlington, Va. Morgan was among four speakers discussing 'Space, the Media and the Millennium' at a reception Jan. 24 kicking off the display at KSC.

  10. Evaluating Education and Science in the KSC Visitor Complex Exhibits

    NASA Technical Reports Server (NTRS)

    Erickson, Lance K.

    2000-01-01

    The continuing development of exhibits at the Kennedy Space Center's Visitor Complex is an excellent opportunity for NASA personnel to promote science and provide insight into NASA programs and projects for the approximately 3 million visitors that come to KSC annually. Stated goals for the Visitor Complex, in fact, emphasize science awareness and recommend broadening the appeal of the displays and exhibits for all age groups. To this end, this summer project seeks to evaluate the science content of planned exhibits/displays in relation to these developing opportunities and identify specific areas for enhancement of existing or planned exhibits and displays. To help expand the educational and science content within the developing exhibits at the Visitor Complex, this project was structured to implement the goals of the Visitor Center Director. To accomplish this, the exhibits and displays planned for completion within the year underwent review and evaluation for science content and educational direction. Planning emphasis for the individual displays was directed at combining the elements of effective education with fundamental scientific integrity, within an appealing format.

  11. University / Science Center Exhibit Development Collaboration: Strategies and Lessons Learned

    NASA Astrophysics Data System (ADS)

    Raddick, M. J.; Carliles, S.; Bartelme, L.; Patterson, J.

    2008-06-01

    Through funding from the NSF's Internship in Public Science Education (IPSE) program, Johns Hopkins University (JHU) and the Maryland Science Center (MSC) have worked together to create an exhibit based on JHU's research with the Sloan Digital Sky Survey, a project to map the universe. The exhibit is a kiosk-based interactive presentation that connects to online data about the sky. It is currently displayed in SpaceLink, an area at the MSC that focuses on current events and research in astronomy. The person primarily responsible for the exhibit was a graduate student in computer science in the JHU Physics and Astronomy department. He worked with an EPO professional in the department and two members of the MSC's planetarium and exhibit staff to plan the exhibit. The team also worked with a coordinator in the JHU chemistry department, and an external evaluator. Along with increased public understanding of science, our goal was to create and evaluate a sustainable partnership between a research university and a local science center. We are producing an evaluation report discussing our collaboration and detailing lessons learned. We hope that our experience can be a model for other university / science center collaborations in the future. Some lessons that we have learned in our development effort are: start all design decisions with learning goals and objectives, write goals with evaluation in mind, focus on the process of science, and do not underestimate the challenges of working with the web as part of the exhibit technology.

  12. A phenomenological investigation of science center exhibition developers' expertise development

    NASA Astrophysics Data System (ADS)

    Young, Denise L.

    The purpose of this study was to examine the exhibition developer role in the context of United States (U.S.) science centers, and more specifically, to investigate the way science center exhibition developers build their professional expertise. This research investigated how successfully practicing exhibition developers described their current practices, how they learned to be exhibition developers, and what factors were the most important to the developers in building their professional expertise. Qualitative data was gathered from 10 currently practicing exhibition developers from three science centers: the Exploratorium, San Francisco, California; the Field Museum, Chicago, Illinois; and the Science Museum of Minnesota, St. Paul, Minnesota. In-depth, semistructured interviews were used to collect the data. The study embraced aspects of the phenomenological tradition and sought to derive a holistic understanding of the position and how expertise was built for it. The data were methodically coded and organized into themes prior to analysis. The data analysis found that the position consisted of numerous and varied activities, but the developers' primary roles were advocating for the visitor, storytelling, and mediating information and ideas. They conducted these activities in the context of a team and relied on an established exhibition planning process to guide their work. Developers described a process of learning exhibition development that was experiential in nature. Learning through daily practice was key, though they also consulted with mentors and relied on visitor studies to gauge the effectiveness of their work. They were adept at integrating prior knowledge gained from many aspects of their lives into their practice. The developers described several internal factors that contributed to their expertise development including the desire to help others, a natural curiosity about the world, a commitment to learning, and the ability to accept critique. They

  13. Black Holes Traveling Exhibition: This Time, It's Personal.

    NASA Astrophysics Data System (ADS)

    Dussault, Mary E.; Braswell, E. L.; Sunbury, S.; Wasser, M.; Gould, R. R.

    2012-01-01

    How can you make a topic as abstract as black holes seem relevant to the life of the average museum visitor? In 2009, the Harvard-Smithsonian Center for Astrophysics developed a 2500 square foot interactive museum exhibition, "Black Holes: Space Warps & Time Twists,” with funding from the National Science Foundation and NASA. The exhibition has been visited by more than a quarter million museum-goers, and is about to open in its sixth venue at the Reuben H. Fleet Science Center in San Diego, California. We have found that encouraging visitors to adopt a custom black hole explorer's identity can help to make the science of black holes more accessible and meaningful. The Black Holes exhibition uses networked exhibit technology that serves to personalize the visitor experience, to support learning over time including beyond the gallery, and to provide a rich quantitative source of embedded evaluation data. Visitors entering the exhibition create their own bar-coded "Black Holes Explorer's Card” which they use throughout the exhibition to collect and record images, movies, their own predictions and conclusions, and other black hole artifacts. This digital database of personal discoveries grows as visitors navigate through the gallery, and an automated web-content authoring system creates a personalized online journal of their experience that they can access once they get home. We report here on new intriguing results gathered from data generated by 112,000 visitors across five different venues. For example, an initial review of the data reveals correlations between visitors’ black hole explorer identity choices and their engagement with the exhibition. We will also discuss correlations between learning gains and personalization.

  14. Space Research Institute (IKI) Exhibition as an Educational Project

    NASA Astrophysics Data System (ADS)

    Sadovski, Andrei; Antonenko, Elena

    2016-07-01

    The Exhibition "Space Science: Part and Future" in Space Research Institute (IKI) was opened in 2007 in commemoration of the 50th anniversary of the first man-made satellite launch. It covers the latest and the most important findings in space research, shows instruments which are used in space exploration, and presents past, current, and future Russian science missions. Prototypes of space instruments developed by Russian specialists and mockups of spacecraft and spaceships flown to space are displayed, together with information posters, describing space missions, their purposes and results. The Exhibition takes a great part in school space education. Its stuff actively works with schoolchildren, undergraduate students and also makes a great contribution in popularization of space researches. Moreover the possibility to learn about scientific space researches first-hand is priceless. We describe the main parts of the Exhibition and forms of it work and also describe the collaboration with other museums and educational organizations.

  15. Liberty Bell 7 Space Capsule Exhibit previews at Visitor Complex

    NASA Technical Reports Server (NTRS)

    2000-01-01

    Children on a tour at the KSC Visitor Complex get an early look at the Discovery Channel's Liberty Bell 7 Space Capsule Exhibit, which opens to the public on Saturday, June 17. They are on a re- creation of the deck of Ocean Project, the ship that located and recovered the space capsule from the floor of the Atlantic Ocean. Liberty Bell 7 launched U.S. Air Force Captain Virgil '''Gus''' Grissom July 21, 1961, on a mission that lasted 15 minutes and 37 seconds before sinking. It lay undetected for nearly four decades before a Discovery Channel expedition located it and recovered it. The space capsule, now restored and preserved, is part of an interactive exhibit touring science centers and museums in 12 cities throughout the United States until 2003. The exhibit also includes hands-on elements such as a capsule simulator, a centrifuge, and ROV pilot.

  16. View of Soyuz spacecraft which was part of exhibit

    NASA Technical Reports Server (NTRS)

    1973-01-01

    A close-up view of the Soyuz spacecraft which was part of the Apollo Soyuz Test Project (ASTP) exhibit at the 30th International Aeronautics and Space Exhibition held May 24 - June 3, 1973 at the Le Bourget Airport in Paris, France. At the far left a mock-up of a Docking Module connects the Apollo spacecraft (not visible) with the Soyuz. The spherical-shaped portion of the Soyuz is called the orbital section. The middle section with the lettering 'CCCP' (USSR) on it is called the Cosmonauts cabin. Two solar panels extend out from the machine and panel section.

  17. Mouse embryonic fibroblasts exhibit extensive developmental and phenotypic diversity.

    PubMed

    Singhal, Prabhat K; Sassi, Slim; Lan, Lan; Au, Patrick; Halvorsen, Stefan C; Fukumura, Dai; Jain, Rakesh K; Seed, Brian

    2016-01-01

    Analysis of embryonic fibroblasts from GFP reporter mice indicates that the fibroblast cell type harbors a large collection of developmentally and phenotypically heterogeneous subtypes. Some of these cells exhibit multipotency, whereas others do not. Multiparameter flow cytometry analysis shows that a large number of distinct populations of fibroblast-like cells can be found in cultures initiated from different embryonic organs, and cells sorted according to their surface phenotype typically retain their characteristics on continued propagation in culture. Similarly, surface phenotypes of individual cloned fibroblast-like cells exhibit significant variation. The fibroblast cell class appears to contain a very large number of denumerable subtypes. PMID:26699463

  18. One exhibition, many goals. Combining scientific research and risk communication

    NASA Astrophysics Data System (ADS)

    Charrière, Marie; Bogaard, Thom; Junier, Sandra; Malet, Jean-Philippe; Mostert, Erik

    2015-04-01

    How effective is visual communication to increase awareness of natural hazards and risks? To answer this research question, we developed a research design that was at the same time an experimental setting and an actual communication effort. Throughout the full length of the 2-years project held in the Ubaye valley (southeastern France), we collaborated with local and regional stakeholders (politicians and technicians). During a consultation phase, the communication context was determined, the audience of the project was defined and finally the testing activity-communication effort was determined. We were offered the opportunity to design an exhibition for the local public library. In a consultation phase that corresponded to the design of the exhibition, the stakeholders contributed to its content as well as helping with the funding of the exhibition. Finally, during the experimentation phase, the stakeholders participated in advertising the activity, gathering of participants and designing the scientific survey. In order to assess the effects of the exhibition on risk awareness, several groups of children, teenagers and adults were submitted to a research design, consisting of 1) a pre-test, 2) the visit of the exhibition and 3) a post-test similar to the pre-test. In addition, the children answered a second post-test 3 months after the visit. Close ended questions addressed the awareness indicators mentioned in the literature, i.e. worry level, previous experiences with natural hazards events, exposure to awareness raising, ability to mitigate/respond/prepare, attitude to risk, and demographics. In addition, the post-test included several satisfaction questions concerning the visual tools displayed in the exhibition. A statistical analysis of the changes between the pre- and post- tests (paired t-test, Wilcoxon signed-rank test and bootstrapping) allowed to verify whether the exhibition had an impact on risk awareness or not. In order to deduce which variable

  19. 13. VIEW OF RAILROAD EXHIBIT AT EL PORTAL. SHAY LOCOMOTIVE ...

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

    13. VIEW OF RAILROAD EXHIBIT AT EL PORTAL. SHAY LOCOMOTIVE IS FROM THE HETCH HETCHY RAILROAD. CABOOSE IS FROM THE YOSEMITE VALLEY RAILROAD. FOREST ROAD IN FOREGROUND IS THE ALIGNMENT OF THE YOSEMITE VALLEY RAILROAD. LOOKING W. GIS: N-37 40 27.0 / W-119 47 10.5 - Yosemite National Park Roads & Bridges, Yosemite Village, Mariposa County, CA

  20. (Un)Disturbing Exhibitions: Indigenous Historical Memory at the NMAI

    ERIC Educational Resources Information Center

    Carpio, Myla Vicenti

    2006-01-01

    Museums in particular are educational tools used to create and perpetuate specific ideologies and historical memories. They have played a prominent role in defining the visibility of Indigenous peoples and cultures in America historical memory by creating exhibits of Indigenous peoples based on perceptions and views that benefit and justify…

  1. 14 CFR Appendix to Subpart A of... - Listing of Exhibits

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 5 2012-01-01 2012-01-01 false Listing of Exhibits Appendix to Subpart A of Part 1260 Aeronautics and Space NATIONAL AERONAUTICS AND SPACE ADMINISTRATION GRANTS AND COOPERATIVE AGREEMENTS General Pt. 1260, Subpt. A, App. Appendix to Subpart A of Part 1260—Listing of...

  2. 14 CFR Appendix to Subpart A of... - Listing of Exhibits

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 5 2013-01-01 2013-01-01 false Listing of Exhibits Appendix to Subpart A of Part 1260 Aeronautics and Space NATIONAL AERONAUTICS AND SPACE ADMINISTRATION GRANTS AND COOPERATIVE AGREEMENTS General Pt. 1260, Subpt. A, App. Appendix to Subpart A of Part 1260—Listing of...

  3. 14 CFR Appendix to Subpart A of... - Listing of Exhibits

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 5 2014-01-01 2014-01-01 false Listing of Exhibits Appendix to Subpart A of Part 1260 Aeronautics and Space NATIONAL AERONAUTICS AND SPACE ADMINISTRATION GRANTS AND COOPERATIVE AGREEMENTS General Pt. 1260, Subpt. A, App. Appendix to Subpart A of Part 1260—Listing of...

  4. A Zn based coordination polymer exhibiting long-lasting phosphorescence.

    PubMed

    Cepeda, Javier; Sebastian, Eider San; Padro, Daniel; Rodríguez-Diéguez, Antonio; García, Jose A; Ugalde, Jesus M; Seco, Jose M

    2016-07-01

    A new Zn(ii) based coordination polymer (CP) built by the cohesive pilling of 2D Shubnikov type layers is reported. This material exhibits time dependent multicoloured emission, part of which shows a persistent green phosphorescence visible for up to two seconds to the naked eye, which originates from multiple charge transfer mechanisms. PMID:27297330

  5. 7 CFR Exhibit B to Subpart B of... - Servicing Company

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... of Agriculture, 7 CFR 2.23; delegation of authority by the Assistant Secretary for Rural Development, 7 CFR 2.70) ... 7 Agriculture 12 2012-01-01 2012-01-01 false Servicing Company B Exhibit B to Subpart B of...

  6. 75 FR 3862 - Photography in Public Exhibit Space

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-01-25

    ..., 2009, NARA published a proposed rule in the Federal Register (74 FR 38153) for a 60-day public comment... RECORDS ADMINISTRATION 36 CFR Part 1280 RIN 3095-AB60 Photography in Public Exhibit Space AGENCY: National... documents accessible and available to the public, and that by prohibiting photography, NARA will make...

  7. Exhibition of Humanities Portfolio with a Latino Urban Population.

    ERIC Educational Resources Information Center

    Estrada, Christelle L. Martinez

    1995-01-01

    Pasadena (California) High School's Puente Pilot Project encourages Hispanic students to pursue college and return home as leaders and mentors. A bilingual counselor, community liaison, and English teacher engage students in an integrated curriculum relevant to their life experiences. Portfolio exhibitions involving oral presentation and…

  8. Peasants on display: the Czechoslavic Ethnographic Exhibition of 1895.

    PubMed

    Filipová, Marta

    2011-01-01

    In the increasingly modernized Central Europe of the late nineteenth century, folk culture, with its alleged ancient character, was still understood by some scholars as the bearer of national identity. The Czechoslavic [sic] Ethnographic Exhibition, which took place in Prague in 1895, aimed to promote the idea of the ethnically unified, but at the same time regionally diverse, identity of the Czech-speaking people living in Bohemia, Moravia and Slovakia. Having to negotiate their identity with the ethnic Germans of Bohemia, the Czechs consciously excluded them from the event both as organizers and as exhibitors. The exhibition could therefore be seen as a symptom of its time—in the late nineteenth century Central Europe, locating national heritage was crucial and folk culture played an important role in the national politics, and not only for the Czechs. This article focuses mainly on the ethnographic exhibit entitled ‘the Exhibition Village’, which consisted of an eclectic selection of village houses and their imitations from Bohemia, Moravia and Slovakia. On this basis, it explores the political intentions behind the display of folk culture to both urban and rural audiences and brings attention to the question of integration of the diverse regional objects in a utopian national whole. The article thus also aims to demonstrate issues related to the use of folk artefacts for the purposes of cultural nationalism in Austria-Hungary in the late nineteenth century. PMID:21574287

  9. Dream-Makers: A National Exhibition of Children's Art.

    ERIC Educational Resources Information Center

    Williams, Susan

    1984-01-01

    The Crayola Dream-Makers exhibition, which consists of 100 two- and three-dimensional works of art in which second, third, and fourth graders from all over the United States depict their dreams for themselves and their world is described. Samples of the children's art are included. (RM)

  10. 12 CFR 625.11 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... regarding release of information (12 CFR part 602). ... 12 Banks and Banking 6 2010-01-01 2010-01-01 false Net worth exhibit. 625.11 Section 625.11 Banks... EXPENSES UNDER THE EQUAL ACCESS TO JUSTICE ACT Applicant Information Required § 625.11 Net worth...

  11. 24 CFR 14.205 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ..., 24 CFR part 15. In either case, disclosure shall be subject to the provisions of the Privacy Act of 1974, 5 U.S.C. 552a, and the Department's procedures implementing the Privacy Act of 1974 at 24 CFR... 24 Housing and Urban Development 1 2010-04-01 2010-04-01 false Net worth exhibit. 14.205...

  12. 19 CFR 212.11 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...'s established procedures under the Freedom of Information Act, 19 CFR 201.17-201.21. ... 19 Customs Duties 3 2010-04-01 2010-04-01 false Net worth exhibit. 212.11 Section 212.11 Customs... IMPLEMENTATION OF THE EQUAL ACCESS TO JUSTICE ACT Information Required From Applicants § 212.11 Net worth...

  13. AERIAL OF VISITORS INFORMATION CENTER [VIC] & ROCKET GARDEN EXHIBIT

    NASA Technical Reports Server (NTRS)

    1975-01-01

    AERIAL OF VISITORS INFORMATION CENTER [VIC] & ROCKET GARDEN EXHIBIT KSC-375C-0604.12 116-KSC-375C-604.12, P-20220, ARCHIVE-04465 Aerial view of Kennedy Space Center Visitors Information Center looking east-northeastward. New food services building under construction is visible at upper left.

  14. The William Parks exhibition denture--a hidden gem.

    PubMed

    Jordan, Peter

    2014-01-01

    This paper describes a previously unknown denture constructed from hippopotamus ivory. The denture is unusual in that it is anatomically correct in terms of tooth morphology. It is believed that it was a demonstration item for the Great Exhibition of 1851. PMID:24620435

  15. Take To the Streets: Guide To Planning Outdoor, Public Exhibits.

    ERIC Educational Resources Information Center

    Cutting, Jennifer McGregor; And Others

    Placing exhibits in public places provides a unique opportunity to reach a broad non-museum-going audience. It offers marketing and publicity opportunities as well as the potential to develop relationships with agencies and individuals who are stakeholders in the public site. The purpose of this guidebook is to describe the steps in creating an…

  16. 7 CFR Appendix - Exhibits to Subpart E of Part 1951

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 14 2014-01-01 2014-01-01 false Exhibits to Subpart E of Part 1951 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE SERVICE, RURAL UTILITIES SERVICE, AND FARM SERVICE AGENCY, DEPARTMENT OF AGRICULTURE (CONTINUED) PROGRAM REGULATIONS (CONTINUED) SERVICING...

  17. 7 CFR Appendix - Exhibits to Subpart E of Part 1951

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 14 2012-01-01 2012-01-01 false Exhibits to Subpart E of Part 1951 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE SERVICE, RURAL UTILITIES SERVICE, AND FARM SERVICE AGENCY, DEPARTMENT OF AGRICULTURE (CONTINUED) PROGRAM REGULATIONS (CONTINUED) SERVICING...

  18. The Cheapbook: A Compendium of Inexpensive Exhibit Ideas, 1995 Edition.

    ERIC Educational Resources Information Center

    Orselli, Paul, Ed.

    This guide includes complete installation descriptions of 30 exhibits. They include: the adjustable birthday cake, ball-in-tube, Bernoulli Box, chain wave, collapsible truss bridge, double wave device, eddy currents raceway, full-length mirror, geodesic domes, giant magnetic tangrams, harmonic cantilever, hyperboloid of revolution, lifting lever,…

  19. 22 CFR Exhibit B to Part 204 - Assignment

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... of ___, ___ the “Guaranty”), between the Lender and the United States of America, acting through the... payment to the undersigned as of the date hereof. By Name Title Dated Accepted: UNITED STATES OF AMERICA.... B Exhibit B to Part 204—Assignment The undersigned, being the registered owner of a Note in...

  20. 22 CFR Exhibit B to Part 204 - Assignment

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... of ___, ___ the “Guaranty”), between the Lender and the United States of America, acting through the... payment to the undersigned as of the date hereof. By Name Title Dated Accepted: UNITED STATES OF AMERICA.... B Exhibit B to Part 204—Assignment The undersigned, being the registered owner of a Note in...

  1. 22 CFR Exhibit B to Part 204 - Assignment

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... of ___, ___ the “Guaranty”), between the Lender and the United States of America, acting through the... payment to the undersigned as of the date hereof. By Name Title Dated Accepted: UNITED STATES OF AMERICA.... B Exhibit B to Part 204—Assignment The undersigned, being the registered owner of a Note in...

  2. 22 CFR Exhibit B to Part 204 - Assignment

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... of ___, ___ the “Guaranty”), between the Lender and the United States of America, acting through the... payment to the undersigned as of the date hereof. By Name Title Dated Accepted: UNITED STATES OF AMERICA.... B Exhibit B to Part 204—Assignment The undersigned, being the registered owner of a Note in...

  3. 22 CFR Exhibit B to Part 204 - Assignment

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... of ___, ___ the “Guaranty”), between the Lender and the United States of America, acting through the... payment to the undersigned as of the date hereof. By Name Title Dated Accepted: UNITED STATES OF AMERICA.... B Exhibit B to Part 204—Assignment The undersigned, being the registered owner of a Note in...

  4. 7 CFR Appendix - Exhibits to Subpart E of Part 1951

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 14 2010-01-01 2009-01-01 true Exhibits to Subpart E of Part 1951 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE SERVICE, RURAL UTILITIES SERVICE, AND FARM SERVICE AGENCY, DEPARTMENT OF AGRICULTURE (CONTINUED) PROGRAM REGULATIONS (CONTINUED) SERVICING...

  5. 7 CFR Appendix - Exhibits to Subpart E of Part 1951

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 14 2011-01-01 2011-01-01 false Exhibits to Subpart E of Part 1951 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE SERVICE, RURAL UTILITIES SERVICE, AND FARM SERVICE AGENCY, DEPARTMENT OF AGRICULTURE (CONTINUED) PROGRAM REGULATIONS (CONTINUED) SERVICING...

  6. 14 CFR Appendix to Subpart A of... - Listing of Exhibits

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 5 2010-01-01 2010-01-01 false Listing of Exhibits Appendix to Subpart A of Part 1260 Aeronautics and Space NATIONAL AERONAUTICS AND SPACE ADMINISTRATION GRANTS AND COOPERATIVE AGREEMENTS General Pt. 1260, Subpt. A, App. Appendix to Subpart A of Part 1260—Listing of...

  7. The "Gravity-Powered Calculator," a Galilean Exhibit

    ERIC Educational Resources Information Center

    Cerreta, Pietro

    2014-01-01

    The Gravity-Powered Calculator is an exhibit of the Exploratorium in San Francisco. It is presented by its American creators as an amazing device that extracts the square roots of numbers, using only the force of gravity. But if you analyze his concept construction one can not help but recall the research of Galileo on falling bodies, the inclined…

  8. Multifunctional materials exhibiting spin crossover and liquid-crystalline properties

    NASA Astrophysics Data System (ADS)

    Seredyuk, M.; Gaspar, Ana B.; Ksenofontov, V.; Reiman, S.; Galyametdinov, Y.; Haase, W.; Rentschler, E.; Gütlich, P.

    2005-11-01

    The physical characterization of a new class of Fe(II) multifunctional SCO materials exhibiting spin crossover and liquid crystalline properties in the room temperatures region is reported. Mössbauer spectroscopy, magnetic, differential scanning calorimetry (DSC), X-ray powder diffraction (XRD) and optical polarizing microscopy studies have been performed on such materials.

  9. Caring for Planet Earth Interactive Exhibit and School Enrichment Program.

    ERIC Educational Resources Information Center

    Kirby, Sarah D.; And Others

    1995-01-01

    The Caring for Planet Earth program addresses concerns and misconceptions of Oklahoma youth about the environment. A school enrichment curriculum reinforces the environmental content of the exhibit at the state fair. Pre-/posttest results verify it is an effective method of educating culturally diverse audiences about environmental issues. (SK)

  10. 7 CFR Exhibit B to Subpart B of... - Servicing Company

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... of Agriculture, 7 CFR 2.23; delegation of authority by the Assistant Secretary for Rural Development, 7 CFR 2.70) ... 7 Agriculture 12 2010-01-01 2010-01-01 false Servicing Company B Exhibit B to Subpart B of...

  11. 17 CFR 230.483 - Exhibits for certain registration statements.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 17 Commodity and Securities Exchanges 2 2010-04-01 2010-04-01 false Exhibits for certain registration statements. 230.483 Section 230.483 Commodity and Securities Exchanges SECURITIES AND EXCHANGE... information as to interest, dividend or conversion rates, redemption or conversion prices, purchase...

  12. 9. COPY OF PHOTOGRAPHIC EXHIBIT BOARD CREATED 19481949 SHOWING CONSTRUCTION ...

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

    9. COPY OF PHOTOGRAPHIC EXHIBIT BOARD CREATED 1948-1949 SHOWING CONSTRUCTION OF ARCH HANGAR. BOARD LOCATED AT AIR FORCE BASE CONVERSION AGENCY, LORING AIR FORCE BASE, MAINE. - Loring Air Force Base, Arch Hangar, East of Arizona Road near southern end of runway, Limestone, Aroostook County, ME

  13. 45 CFR 1160.5 - Eligibility for domestic exhibitions.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 45 Public Welfare 3 2011-10-01 2011-10-01 false Eligibility for domestic exhibitions. 1160.5 Section 1160.5 Public Welfare Regulations Relating to Public Welfare (Continued) NATIONAL FOUNDATION ON THE ARTS AND THE HUMANITIES FEDERAL COUNCIL ON THE ARTS AND THE HUMANITIES INDEMNITIES UNDER THE...

  14. 18 CFR 157.16 - Exhibits relating to acquisitions.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... purchase, sale, or interchange of natural gas, and upon any rate schedules or tariffs on file with this... required to be made with this Commission. (b) Exhibit R—Acquisition contracts. A summary statement of all..., if the acquisition involved is by purchase of capital stock and liquidation of the acquired...

  15. Curriculum Activities Guide for Natural History Exhibits, Grades K-8.

    ERIC Educational Resources Information Center

    International Wildlife Museum, Tucson, AZ.

    A natural history museum is a building where animals, plants, minerals, and other things in nature are kept and exhibited for study. This document is a curriculum guide to provide a variety of activities for educators and their students to use not only when visiting the International Wildlife Museum (Tuscon, Arizona), but also with natural history…

  16. 40 CFR 17.12 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 1 2011-07-01 2011-07-01 false Net worth exhibit. 17.12 Section 17.12 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GENERAL IMPLEMENTATION OF THE EQUAL ACCESS TO JUSTICE ACT IN EPA ADMINISTRATIVE PROCEEDINGS Information Required From Applicants § 17.12 Net...

  17. 40 CFR 17.12 - Net worth exhibit.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 1 2012-07-01 2012-07-01 false Net worth exhibit. 17.12 Section 17.12 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GENERAL IMPLEMENTATION OF THE EQUAL ACCESS TO JUSTICE ACT IN EPA ADMINISTRATIVE PROCEEDINGS Information Required From Applicants § 17.12 Net...

  18. 40 CFR 17.12 - Net worth exhibit.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 1 2014-07-01 2014-07-01 false Net worth exhibit. 17.12 Section 17.12 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GENERAL IMPLEMENTATION OF THE EQUAL ACCESS TO JUSTICE ACT IN EPA ADMINISTRATIVE PROCEEDINGS Information Required From Applicants § 17.12 Net...

  19. 40 CFR 17.12 - Net worth exhibit.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 1 2013-07-01 2013-07-01 false Net worth exhibit. 17.12 Section 17.12 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY GENERAL IMPLEMENTATION OF THE EQUAL ACCESS TO JUSTICE ACT IN EPA ADMINISTRATIVE PROCEEDINGS Information Required From Applicants § 17.12 Net...

  20. Early Childhood Commercial Exhibit Controversies: 1890 and 1990.

    ERIC Educational Resources Information Center

    Hewes, Dorothy W.

    The controversy over the contrast between early childhood principles and the commercial exhibits at the 1990 annual meeting of the National Association for the Education of Young Children prompted a search for historical antecedents. Research disclosed that the first popular linking of Froebelian kindergarten curriculum and manufactured equipment…

  1. Do Online Learning Patterns Exhibit Regional and Demographic Differences?

    ERIC Educational Resources Information Center

    Hsieh, Tsui-Chuan; Yang, Chyan

    2012-01-01

    This paper used a multi-level latent class model to evaluate whether online learning patterns exhibit regional differences and demographics. This study discovered that the Internet learning pattern consists of five segments, and the region of Taiwan is divided into two segments and further found that both the user and the regional segments are…

  2. Explanatory Parent-Child Conversation Predominates at an Evolution Exhibit

    ERIC Educational Resources Information Center

    Tare, Medha; French, Jason; Frazier, Brandy N.; Diamond, Judy; Evans, E. Margaret

    2011-01-01

    To investigate how parents support children's learning at an exhibit on evolution, the conversations of 12 families were recorded, transcribed, and coded (6,263 utterances). Children (mean age 9.6 years) and parents visited Explore Evolution, which conveyed current research about the evolution of seven organisms. Families were engaged with the…

  3. 17 CFR 229.601 - (Item 601) Exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... specific exhibit filing requirements applicable thereto. (4) If a material contract or plan of acquisition... forms 10 8-K 2 10-D 10-Q 10-K (1) Underwriting agreement X X X X X X X X (2) Plan of acquisition... into a registration statement subsequent to its effectiveness. (2) Plan of acquisition,...

  4. 14 CFR Appendix to Part 1274 - Listing of Exhibits

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 5 2011-01-01 2010-01-01 true Listing of Exhibits Appendix to Part 1274 Aeronautics and Space NATIONAL AERONAUTICS AND SPACE ADMINISTRATION COOPERATIVE AGREEMENTS WITH COMMERCIAL... Order 11246 Relating to Equal Employment Opportunity,” and as supplemented by regulations at 41 CFR...

  5. Brainstorming Interactive Exhibits for the Baseball Hall of Fame.

    ERIC Educational Resources Information Center

    Shapiro, David W.; Seidman, Steven

    1996-01-01

    Discusses incorporating interactive multimedia into the Baseball Hall of Fame in Cooperstown, New York. Presents examples of interactive exhibits from other museums and suggests ideas for the Hall of Fame to pursue: simulations of batting, signal use, and umpiring; a quiz show; a visitor poll; CD-ROM software; and an interactive database.…

  6. 45 CFR 1160.5 - Eligibility for domestic exhibitions.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 45 Public Welfare 3 2012-10-01 2012-10-01 false Eligibility for domestic exhibitions. 1160.5 Section 1160.5 Public Welfare Regulations Relating to Public Welfare (Continued) NATIONAL FOUNDATION ON THE ARTS AND THE HUMANITIES FEDERAL COUNCIL ON THE ARTS AND THE HUMANITIES INDEMNITIES UNDER THE...

  7. 45 CFR 1160.5 - Eligibility for domestic exhibitions.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 45 Public Welfare 3 2013-10-01 2013-10-01 false Eligibility for domestic exhibitions. 1160.5 Section 1160.5 Public Welfare Regulations Relating to Public Welfare (Continued) NATIONAL FOUNDATION ON THE ARTS AND THE HUMANITIES FEDERAL COUNCIL ON THE ARTS AND THE HUMANITIES INDEMNITIES UNDER THE...

  8. 45 CFR 1160.5 - Eligibility for domestic exhibitions.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 45 Public Welfare 3 2014-10-01 2014-10-01 false Eligibility for domestic exhibitions. 1160.5 Section 1160.5 Public Welfare Regulations Relating to Public Welfare (Continued) NATIONAL FOUNDATION ON THE ARTS AND THE HUMANITIES FEDERAL COUNCIL ON THE ARTS AND THE HUMANITIES INDEMNITIES UNDER THE...

  9. 63. View of Klystron tube cutaway exhibit located at mezzanine ...

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

    63. View of Klystron tube cut-away exhibit located at mezzanine level transmitter building no. 102, directly above RF power generation systems located on first floor. - Clear Air Force Station, Ballistic Missile Early Warning System Site II, One mile west of mile marker 293.5 on Parks Highway, 5 miles southwest of Anderson, Anderson, Denali Borough, AK

  10. Conversational Competency Profiles of Adult Males Who Exhibit Fluency Disorders.

    ERIC Educational Resources Information Center

    Thorpe, Carol D.

    The study explored the verbal and nonverbal conversational competency of five adult males who exhibited fluency disorders. Five subject/interactant videotaped conversational interactions were analyzed utilizing an INter-REActive Learning (INREAL) Model analysis format. Descriptive individual and composite profiles resulting from trained raters'…

  11. Alien Earths: A Traveling Science Exhibit and Education Program

    NASA Astrophysics Data System (ADS)

    Dusenbery, P. B.; Morrow, C. A.; Harold, J.

    2004-05-01

    Where did we come from? Are we alone? These age-old questions form the basis of NASA's Origins Program, a series of missions spanning the next twenty years that will use a host of space- and ground-based observatories to understand the origin and development of galaxies, stars, planets, and the conditions necessary to support life. The Space Science Institute in Boulder, CO, is developing a 3,000 square-foot traveling exhibition, called Alien Earths, which will bring origins-related research and discoveries to students and the American public. Alien Earths will have four interrelated exhibit areas: Our Place in Space, Star Birth, PlanetQuest, and Search for Life. Exhibit visitors will explore the awesome events surrounding the birth of stars and planets; they will join scientists in the hunt for planets outside our solar system including those that may be in "habitable zones" around other stars; and finally they will be able to learn about the wide range of conditions for life on Earth and how scientists are looking for signs of life beyond Earth. Visitors will also learn about the tools scientists use, such as space-based and ground-based telescopes, to improve our understanding of the cosmos. The exhibit's size will permit it to visit medium sized museums in all regions of the country. It will begin its 3-year tour to 9 host museums and science centers in early 2005 at the Lawrence Hall of Science in Berkeley, California. The Association of Science-Technology Centers (ASTC) will manage the exhibit's national tour. In addition to the exhibit, the project includes workshops for educators and docents at host sites, as well as a public website that will use exhibit content to delve deeper into origins research. Current partners in the Alien Earths project include ASTC, Denver Museum of Nature and Science, Lawrence Hall of Science, NASA Astrobiology Institute, NASA missions (Navigator, SIRTF, and Kepler), the SETI Institute, and the Space Telescope Science Institute

  12. Inside active volcanoes; an exhibit on the move!

    USGS Publications Warehouse

    Fiske, R.S.

    1990-01-01

    All of us are aware of the emphasis currently being placed in the United States on science education and public understanding of science. Most of this emphasis is directed toward mass audiences through book publications, school curricula, and television programs; sadly, most of it deals with non-earth science topics. In an effort to take advantage of this awakened consciousness and to highlight the earth sciences, the Smithsonian Institution and the U.S Geological Survey joined forces to prepare a traveling exhibit on volcanoes that is currently touring the country. This note will serve to bring you up to date on the progress of this exhibit as it reaches the mid-point of its tour. 

  13. The Making of the Fathers of Astronomy Exhibit

    NASA Astrophysics Data System (ADS)

    Graney, C. M.

    2010-10-01

    The International Year of Astronomy 2009 stretched a few days into 2010 here in Louisville, Kentucky - the Fathers of Astronomy exhibit at the Frazier International History Museum did not close until 3 January 2010. Fathers of Astronomy, which was open for five months, told the story of Galileo through authentic original editions of three books - the 1493 Nuremberg Chronicle, Nicolas Copernicus's 1543 On the Revolutions of Heavenly Spheres, and Galileo Galilei's 1632 Dialogue Concerning the two Chief World Systems. The success of "Fathers" resulted from three very different partners coming together and combining resources to produce a history-themed IYA2009 programme of the highest quality at minimal cost. Lessons learned from the exhibit may be of value to people interested in communicating astronomy to the public.

  14. New neptunium(V) borates that exhibit the alexandrite effect.

    PubMed

    Wang, Shuao; Alekseev, Evgeny V; Depmeier, Wulf; Albrecht-Schmitt, Thomas E

    2012-01-01

    A new neptunium(V) borate, K[(NpO(2))B(10)O(14)(OH)(4)], was synthesized using boric acid as a reactive flux. The compound possesses a layered structure in which Np(V) resides in triangular holes, creating a hexagonal-bipyramidal environment around neptunium. This compound is unusual in that it exhibits the Alexandrite effect, a property that is typically restricted to neptunium(IV) compounds.

  15. Environmental enrichment for a mixed-species nocturnal mammal exhibit.

    PubMed

    Clark, Fay E; Melfi, Vicky A

    2012-01-01

    Environmental enrichment (EE) is an integral aspect of modern zoo animal management but, empirical evaluation of it is biased toward species housed in single-species groups. Nocturnal houses, where several nocturnal species are housed together, are particularly overlooked. This study investigated whether three species (nine-banded armadillos, Dasypus novemcinctus; Senegal bush babies, Galago senegalensis; two-toed sloths, Choloepus didactylus) in the nocturnal house at Paignton Zoo Environmental Park, UK could be enriched using food-based and sensory EE. Subjects were an adult male and female of each species. EE was deemed effective if it promoted target species-typical behaviors, behavioral diversity, and increased use of enriched exhibit zones. Results from generalized linear mixed models demonstrated that food-based EE elicited the most positive behavioral effects across species. One set of food-based EEs (Kong®, termite mound and hanging food) presented together was associated with a significant increase in species-typical behaviors, increased behavioral diversity, and increased use of enriched exhibit zones in armadillos and bush babies. Although one type of sensory EE (scented pine cones) increased overall exhibit use in all species, the other (rainforest sounds) was linked to a significant decrease in species-typical behavior in bush babies and sloths. There were no intra or interspecies conflicts over EE, and commensalism occurred between armadillos and bush babies. Our data demonstrate that simple food-based and sensory EE can promote positive behavioral changes in a mixed-species nocturnal mammal exhibit. We suggest that both food and sensory EE presented concurrently will maximize opportunities for naturalistic activity in all species.

  16. Inuit Perspectives on Arctic Environmental Change': A Traveling Exhibition

    NASA Astrophysics Data System (ADS)

    Sheffield, E. M.; Hakala, J. S.; Gearheard, S.

    2006-12-01

    The Inuit of Nunavut, Canada, have an intimate relationship with their surroundings. As a culture that relies on knowledge of sea ice, snow, and weather conditions for success in hunting, fishing, and healthy wellbeing, Inuit have observed and studied environmental patterns for generations. An ongoing study into their traditional knowledge and their observations of environmental change is being conducted by researcher Dr. Shari Gearheard, who has worked with Inuit communities in Nunavut for over a decade. The results of the research have been published in scientific journals, and to communicate the results to a broader audience, Dr. Gearheard designed an interactive CD-ROM displaying photographs, maps, and interview videos of Inuit Elders' perspectives on the changes they have witnessed. Receiving immediate popularity since its release in 2004, copies of `When the Weather is Uggianaqtuq: Inuit Observations of Environmental Change' have been distributed worldwide, to indigenous peoples, social science and climate change researchers, teachers, students, and the general public. To further disseminate the information contained on the CD-ROM, the National Snow and Ice Data Center and the Museum of Natural History, both of the University of Colorado, are partnering to create an exhibition which will open at the Museum during the International Polar Year in April 2008. The exhibit, tentatively titled `Inuit Perspectives on Arctic Environmental Change,' will feature photographs, graphics, and text in both English and Inuktitut describing environmental change in the North. The goals are to make the information and interpretation contained on the CD-ROM available and more accessible to a broad audience and to raise awareness about Arctic climate change and the important contribution of Inuit knowledge. Following exhibition at the Museum, the exhibit will travel throughout the United States, Alaska, and Nunavut, through a network of museums, schools, libraries, tribal

  17. Environmental enrichment for a mixed-species nocturnal mammal exhibit.

    PubMed

    Clark, Fay E; Melfi, Vicky A

    2012-01-01

    Environmental enrichment (EE) is an integral aspect of modern zoo animal management but, empirical evaluation of it is biased toward species housed in single-species groups. Nocturnal houses, where several nocturnal species are housed together, are particularly overlooked. This study investigated whether three species (nine-banded armadillos, Dasypus novemcinctus; Senegal bush babies, Galago senegalensis; two-toed sloths, Choloepus didactylus) in the nocturnal house at Paignton Zoo Environmental Park, UK could be enriched using food-based and sensory EE. Subjects were an adult male and female of each species. EE was deemed effective if it promoted target species-typical behaviors, behavioral diversity, and increased use of enriched exhibit zones. Results from generalized linear mixed models demonstrated that food-based EE elicited the most positive behavioral effects across species. One set of food-based EEs (Kong®, termite mound and hanging food) presented together was associated with a significant increase in species-typical behaviors, increased behavioral diversity, and increased use of enriched exhibit zones in armadillos and bush babies. Although one type of sensory EE (scented pine cones) increased overall exhibit use in all species, the other (rainforest sounds) was linked to a significant decrease in species-typical behavior in bush babies and sloths. There were no intra or interspecies conflicts over EE, and commensalism occurred between armadillos and bush babies. Our data demonstrate that simple food-based and sensory EE can promote positive behavioral changes in a mixed-species nocturnal mammal exhibit. We suggest that both food and sensory EE presented concurrently will maximize opportunities for naturalistic activity in all species. PMID:21387395

  18. New Constitutively Active Phytochromes Exhibit Light-Independent Signaling Activity.

    PubMed

    Jeong, A-Reum; Lee, Si-Seok; Han, Yun-Jeong; Shin, Ah-Young; Baek, Ayoung; Ahn, Taeho; Kim, Min-Gon; Kim, Young Soon; Lee, Keun Woo; Nagatani, Akira; Kim, Jeong-Il

    2016-08-01

    Plant phytochromes are photoreceptors that mediate a variety of photomorphogenic responses. There are two spectral photoisomers, the red light-absorbing Pr and far-red light-absorbing Pfr forms, and the photoreversible transformation between the two forms is important for the functioning of phytochromes. In this study, we isolated a Tyr-268-to-Val mutant of Avena sativa phytochrome A (AsYVA) that displayed little photoconversion. Interestingly, transgenic plants of AsYVA showed light-independent phytochrome signaling with a constitutive photomorphogenic (cop) phenotype that is characterized by shortened hypocotyls and open cotyledons in the dark. In addition, the corresponding Tyr-303-to-Val mutant of Arabidopsis (Arabidopsis thaliana) phytochrome B (AtYVB) exhibited nuclear localization and interaction with phytochrome-interacting factor 3 (PIF3) independently of light, conferring a constitutive photomorphogenic development to its transgenic plants, which is comparable to the first constitutively active version of phytochrome B (YHB; Tyr-276-to-His mutant). We also found that chromophore ligation was required for the light-independent interaction of AtYVB with PIF3. Moreover, we demonstrated that AtYVB did not exhibit phytochrome B activity when it was localized in the cytosol by fusion with the nuclear export signal and that AsYVA exhibited the full activity of phytochrome A when localized in the nucleus by fusion with the nuclear localization signal. Furthermore, the corresponding Tyr-269-to-Val mutant of Arabidopsis phytochrome A (AtYVA) exhibited similar cop phenotypes in transgenic plants to AsYVA. Collectively, these results suggest that the conserved Tyr residues in the chromophore-binding pocket play an important role during the Pr-to-Pfr photoconversion of phytochromes, providing new constitutively active alleles of phytochromes by the Tyr-to-Val mutation. PMID:27325667

  19. Uranium mining wastes, garden exhibition and health risks

    SciTech Connect

    Schmidt, Gerhard; Schmidt, Peter; Hinz, Wilko

    2007-07-01

    Available in abstract form only. Full text of publication follows: For more than 40 years the Soviet-German stockholding company SDAG WISMUT mined and milled Uranium in the East of Germany and became up to 1990 the world's third largest Uranium producer. After reunification of Germany, the new found state own company Wismut GmbH was faced with the task of decommissioning and rehabilitation of the mining and milling sites. One of the largest mining areas in the world, that had to be cleaned up, was located close to the municipality of Ronneburg near the City of Gera in Thuringia. After closing the operations of the Ronneburg underground mine and at the 160 m deep open pit mine with a free volume of 84 Mio.m{sup 3}, the open pit and 7 large piles of mine waste, together 112 Mio.m{sup 3} of material, had to be cleaned up. As a result of an optimisation procedure it was chosen to relocate the waste rock piles back into the open pit. After taking this decision and approval of the plan the disposal operation was started. Even though the transport task was done by large trucks, this took 16 years. The work will be finished in 2007, a cover consisting of 40 cm of uncontaminated material will be placed on top of the material, and the re-vegetation of the former open pit area will be established. When in 2002 the City of Gera applied to host the largest garden exhibition in Germany, Bundesgartenschau (BUGA), in 2007, Wismut GmbH supported this plan by offering parts of the territory of the former mining site as an exhibition ground. Finally, it was decided by the BUGA organizers to arrange its 2007 exhibition on grounds in Gera and in the valley adjacent to the former open pit mine, with parts of the remediated area within the fence of the exhibition. (authors)

  20. Didactical Holographic Exhibit Including Holo TV (holographic Television)

    NASA Astrophysics Data System (ADS)

    Lunazzi, José J.; Magalhães, Daniel S. F.; Rivera, Noemí I. R.

    2008-04-01

    Our Institute of Physics exposes since 1980 didactical exhibitions of holography in Brazil where nice holograms are shown altogether with basic experiments of geometric and wave optics. This experiments lead to the understanding of the phenomenon of images of an ample way. Thousands of people have been present at them, in their majority of the Universidade Estadual de Campinas, where since 2002 they have taken the format of a course without formal evaluation. This way the exhibition has been divided in four modules, in each one of them are shown different holograms, experiments of optics and applications of diffractive images with white light developed in the Institute of Physics. The sequence of the learning through the modules begins with the geometric optics, later we explain the wave optics and finally holography. The phenomenon of the diffraction in daily elements is shown experimentally from the beginning. As well as the application of the holographic screens in white light: the television images that appear in front of the screen and the spectator can try to experience the reality illusion. Put something so exclusive (that only exists in the laboratory) to the public is a way to approximate the persons to an investigation in course. The vision of images that seem to be of holograms, but in movement, and size of until a square meter completes this exhibition of an exclusive way in the world.

  1. Liberty Bell 7 Space Capsule Exhibit previews at Visitor Complex

    NASA Technical Reports Server (NTRS)

    2000-01-01

    Media gather at the KSC Visitor Complex for the kickoff of the Discovery Channel's Liberty Bell 7 Space Capsule Exhibit, which will open to the public on Saturday, June 17. At the podium is Mike Quattrone, executive vice president and general manager, Discovery Channel. Standing to the left of the podium is Rick Abramson, president and chief operating officer of Delaware North Parks Services of Spaceport, Inc., and far left, Jim Jennings, deputy director of Kennedy Space Center. Liberty Bell 7 launched U.S. Air Force Captain Virgil '''Gus''' Grissom July 21, 1961 on a mission that lasted 15 minutes and 37 seconds before sinking to the floor of the Atlantic Ocean, three miles deep. It lay undetected for nearly four decades before a Discovery Channel expedition located it and recovered it. The space capsule is now restored and preserved, and part of an interactive exhibit touring science centers and museums in 12 cities throughout the United States until 2003. The exhibit includes hands-on elements such as a capsule simulator, a centrifuge, and ROV pilot.

  2. Liberty Bell 7 Space Capsule Exhibit previews at Visitor Complex

    NASA Technical Reports Server (NTRS)

    2000-01-01

    The Discovery Channel's Liberty Bell 7 Space Capsule Exhibit, which opens to the public at the KSC Visitor Complex on Saturday, June 17, had a preview for the press today. Liberty Bell 7 launched U.S. Air Force Captain Virgil 'Gus' Grissom July 21, 1961, on a mission that lasted 15 minutes and 37 seconds before sinking. The capsule lay undetected for nearly four decades before a Discovery Channel expedition located it and recovered it. Standing in front of the restored Liberty Bell 7 capsule are (left to right) KSC's Deputy Director Jim Jennings; Gunther Wendt, who worked on the Liberty Bell 7 before its launch; Jim Lewis, who piloted the Hunt Club 1 helicopter that rescued Gus Grissom; and Larry Grissom, brother of Gus Grissom. The space capsule, now restored and preserved, is part of an interactive exhibit touring science centers and museums in 12 cities throughout the United States until 2003. The exhibit also includes hands-on elements such as a capsule simulator, a centrifuge, and ROV pilot.

  3. Environmental Salmonella in agricultural fair poultry exhibits in Colorado.

    PubMed

    Pabilonia, K L; Cadmus, K J; Lingus, T M; Bolte, D S; Russell, M M; Van Metre, D C; Erdman, M M

    2014-03-01

    Salmonella enterica is a common zoonotic pathogen in humans. Transmission typically occurs through consumption of contaminated food products or contact with infected animals, including poultry or their environment. The objective of this study was to estimate the frequency of Salmonella contamination in the environment in poultry exhibits at agricultural fairs. Samples were collected from cages, feed, floors and tables in the exhibit and cultured for Salmonella. At least one environmental sample was positive for Salmonella in 10 of 11 fairs (91%), and Salmonella was isolated from 28 of 55 environmental samples (50.9%). Eleven different serotypes were detected. Results of this study demonstrate that environmental surfaces at agricultural fairs can be contaminated with Salmonella and could potentially serve as a route of transmission to bird owners and the general public. Poultry owners and the general public should be educated about the risks of Salmonella infection from the poultry exhibit environment. Agricultural fairs should consider instituting policies and practices to improve hygiene and mitigate the risk of zoonotic salmonellosis.

  4. Exhibition QIM-based watermarking for digital cinema

    NASA Astrophysics Data System (ADS)

    Callau, Pilar; Darazi, Rony; Macq, Benoît

    2009-02-01

    The copyright protection of Digital Cinema requires the insertion of forensic watermarks during exhibition playback. This paper presents a low-complexity exhibition watermarking method based on quantization index modulation (QIM) and embedded in the DCI compliant decoder. Watermark embedding is proposed to fit in the JPEG2000 decoding process, prior to the inverse wavelet transform and such as it has a minimal impact on the image quality, guarantying a strong link between decompression and watermarking. The watermark is embedded by using an adaptive-Spread Transform Dither Modulation (STDM) method, based on a new multi-resolution perceptual masking to adapt watermark strength. Watermark detection is thereafter performed over the wavelet transformation of the recovered images. The proposed approach offers a wide range of channel capacities according to robustness to several kinds of distortions while maintaining a low computational complexity. Watermarking detection performance on Digital Cinema pictures captured with a video camera from a viewing room has been preliminary assessed, showing very promising results. The proposed approach provides high levels of imperceptibility, yet good robustness to degradations resulting from camcorder exhibition capture, to common signal processing operations such as filtering or re-sampling, and to very high compression.

  5. Electroanalytical determination of tungsten and molybdenum in proteins.

    PubMed

    Hagedoorn, P L; van't Slot, P; van Leeuwen, H P; Hagen, W R

    2001-10-01

    Recent crystal structure determinations accelerated the progress in the biochemistry of tungsten-containing enzymes. In order to characterize these enzymes, a sensitive determination of this metal in protein-containing samples is necessary. An electroanalytical tungsten determination has successfully been adapted to determine the tungsten and molybdenum content in enzymes. The tungsten and molybdenum content can be measured simultaneously from 1 to 10 microg of purified protein with little or no sample handling. More crude protein samples require precipitation of interfering surface active material with 10% perchloric acid. This method affords the isolation of novel molybdenum- and tungsten-containing proteins via molybdenum and tungsten monitoring of column fractions, without using radioactive isotopes. A screening of soluble proteins from Pyrococcus furiosus for tungsten, using anion-exchange column chromatography to separate the proteins, has been performed. The three known tungsten-containing enzymes from P. furiosus were recovered with this screening.

  6. Human Myocardial Pericytes: Multipotent Mesodermal Precursors Exhibiting Cardiac Specificity

    PubMed Central

    Chen, William C.W.; Baily, James E.; Corselli, Mirko; Diaz, Mary; Sun, Bin; Xiang, Guosheng; Gray, Gillian A.; Huard, Johnny; Péault, Bruno

    2015-01-01

    Perivascular mesenchymal precursor cells (i.e. pericytes) reside in skeletal muscle where they contribute to myofiber regeneration; however, the existence of similar microvessel-associated regenerative precursor cells in cardiac muscle has not yet been documented. We tested whether microvascular pericytes within human myocardium exhibit phenotypes and multipotency similar to their anatomically and developmentally distinct counterparts. Fetal and adult human heart pericytes (hHPs) express canonical pericyte markers in situ, including CD146, NG2, PDGFRβ, PDGFRα, αSMA, and SM-MHC, but not CD117, CD133 and desmin, nor endothelial cell (EC) markers. hHPs were prospectively purified to homogeneity from ventricular myocardium by flow cytometry, based on a combination of positive- (CD146) and negative-selection (CD34, CD45, CD56, and CD117) cell lineage markers. Purified hHPs expanded in vitro were phenotypically similar to human skeletal muscle-derived pericytes (hSkMPs). hHPs express MSC markers in situ and exhibited osteo- chondro-, and adipogenic potentials but, importantly, no ability for skeletal myogenesis, diverging from pericytes of all other origins. hHPs supported network formation with/without ECs in Matrigel cultures; hHPs further stimulated angiogenic responses under hypoxia, markedly different from hSkMPs. The cardiomyogenic potential of hHPs was examined following 5-azacytidine treatment and neonatal cardiomyocyte co-culture in vitro, and intramyocardial transplantation in vivo. Results indicated cardiomyocytic differentiation in a small fraction of hHPs. In conclusion, human myocardial pericytes share certain phenotypic and developmental similarities with their skeletal muscle homologs, yet exhibit different antigenic, myogenic, and angiogenic properties. This is the first example of an anatomical restriction in the developmental potential of pericytes as native mesenchymal stem cells. PMID:25336400

  7. Homogeneous illusion device exhibiting transformed and shifted scattering effect

    NASA Astrophysics Data System (ADS)

    Mei, Jin-Shuo; Wu, Qun; Zhang, Kuang; He, Xun-Jun; Wang, Yue

    2016-06-01

    Based on the theory of transformation optics, a type of homogeneous illusion device exhibiting transformed and shifted scattering effect is proposed in this paper. The constitutive parameters of the proposed device are derived, and full-wave simulations are performed to validate the electromagnetic properties of transformed and shifted scattering effect. The simulation results show that the proposed device not only can visually shift the image of target in two dimensions, but also can visually transform the shape of target. It is expected that such homogeneous illusion device could possess potential applications in military camouflage and other field of electromagnetic engineering.

  8. Predicting supercontinuum pulse collisions with simulations exhibiting temporal aliasing.

    PubMed

    Liu, Chu; Rees, Eric J; Laurila, Toni; Jian, Shuisheng; Kaminski, Clemens F

    2010-12-15

    Interactions between supercontinuum (SC) light pulses, produced by the propagation of rapidly sequenced picosecond pump laser pulses along a photonic crystal fiber, result in spectral broadening, which we attribute to interpulse soliton collisions. This phenomenon was measured experimentally, following our observation of spectral broadening in numerical simulations that exhibit so-called "pulse wraparound" or "temporal aliasing." This occurs in simulations with narrow time grids: as early parts of the SC pulse leave the computational time domain, they "reenter" at the beginning and so interact with later parts of the evolving SC pulse. We show that this provides an effective model to predict the experimentally observed spectral changes.

  9. An application of projection imaging systems for museum exhibitions

    NASA Astrophysics Data System (ADS)

    Miyata, Kimiyoshi; Takiguchi, Takahiro; Nakaguchi, Toshiya; Tsumura, Norimichi; Miyake, Yoichi

    2008-01-01

    Museums hold a wide variety of cultural properties and they organize exhibitions for the public. In history museums, a number of interpretive materials are necessary to present a specific historical theme. However, due to limitation of space, the number of displayed materials is severely restricted. There are various types of guidance systems in museums, however, most systems have not yet responded to visitors' needs. Therefore, a useful guidance system is necessary to satisfy their needs and also to enhance their understandings of the exhibitions. In order to create such guidance system, applications of imaging technology can become a solution to overcome these restrictions and meet the requirements. In this research, a visual guidance system using a data projector was examined in the National Museum of Japanese History. In the kimono (traditional Japanese clothing) section, the projector was used to simulate color changes of the materials under different illuminants. The projector also highlighted areas where annotations were featured in the kimono. The validity of using the projector system was confirmed by results from an interview survey. To further develop this visual guidance system, an augmented reality system consisted of cooperation between the projector and a digital camera was also examined.

  10. Endophytic Fungi Isolated from Coleus amboinicus Lour Exhibited Antimicrobial Activity

    PubMed Central

    Astuti, Puji; Sudarsono, Sudarsono; Nisak, Khoirun; Nugroho, Giri Wisnu

    2014-01-01

    Purpose: Coleus amboinicus is a medicinal plant traditionally used to treat various diseases such as throat infection, cough and fever, diarrhea, nasal congestion and digestive problems. The plant was explored for endophytic fungi producing antimicrobial agents. Methods: Screening for endophytic fungi producing antimicrobial agents was conducted using agar plug method and antimicrobial activity of promising ethyl acetate extracts was determined by disc diffusion assay. Thin layer chromatography (TLC) - bioautography was performed to localize the bioactive components within the extract. TLC visualization detection reagents were used to preliminary analyze phytochemical groups of the bioactive compounds. Results: Three endophytic fungi were obtained, two of them showed promising potential. Agar diffusion method showed that endophytic fungi CAL-2 exhibited antimicrobial activity against P. aeruginosa, B. subtilis, S. aureus and S. thypi, whilst CAS-1 inhibited the growth of B. subtilis. TLC bioautography of ethyl acetate extract of CAL-2 revealed at least three bands exhibited antimicrobial activity and at least two bands showed inhibition of B. subtilis growth. Preliminary analysis of the crude extracts suggests that bioactive compounds within CAL-2 extract are terpenoids, phenolics and phenyl propanoid compounds whilst the antimicrobial agents within CAS-1 extract are terpenoids, propylpropanoids, alkaloids or heterocyclic nitrogen compounds. Conclusion: These data suggest the potential of endophytic fungi of C. amboinicus as source for antimicrobial agents. PMID:25671195

  11. Parallax Park - A bilingual, outdoor, interactive family exhibit

    NASA Astrophysics Data System (ADS)

    Hemenway, M. K.; Benedict, G. F.; Preston, S. L.; Armosky, B. J.; Cianciolo, F. W., Jr.; Wetzel, M. N.; Freeman, R.; Ransenberg, A.

    2004-12-01

    We present a design for Parallax Park, an outdoor, interactive family exhibit for McDonald Observatory that will bring the quest for learning about extra-solar planets and the astronomical distance ladder down to Earth for the more than 100,000 visitors who come to the Observatory each year. The Park will teach visitors the basic principles of astrometry. It will concentrate on distance determination and extrasolar planet detection, and emphasize the advantages of space-based astrometric measurements. Visitors will experience parallax by traveling a path around a representation of the Sun, mimicking Earth's orbit, while viewing representations of stars at various distances and in various directions. Concepts include Cepheids as standard candles, the inverse square law, and motion around a center of mass. The exhibit includes interactive components suitable for children's use. Text labels and printed guides, in English and Spanish, will explain how to use the Park to explore the uses of astrometry in modern science. This program was made possible through funding from the Hubble Space Telescope Cycle 11 and 12 Education and Public Outreach Grant Programs. Support from the NASA/JPL Space Interferometry Mission (JPL contract #1227563) is also gratefully acknowledged.

  12. Chimpanzees and bonobos exhibit divergent spatial memory development.

    PubMed

    Rosati, Alexandra G; Hare, Brian

    2012-11-01

    Spatial cognition and memory are critical cognitive skills underlying foraging behaviors for all primates. While the emergence of these skills has been the focus of much research on human children, little is known about ontogenetic patterns shaping spatial cognition in other species. Comparative developmental studies of nonhuman apes can illuminate which aspects of human spatial development are shared with other primates, versus which aspects are unique to our lineage. Here we present three studies examining spatial memory development in our closest living relatives, chimpanzees (Pan troglodytes) and bonobos (P. paniscus). We first compared memory in a naturalistic foraging task where apes had to recall the location of resources hidden in a large outdoor enclosure with a variety of landmarks (Studies 1 and 2). We then compared older apes using a matched memory choice paradigm (Study 3). We found that chimpanzees exhibited more accurate spatial memory than bonobos across contexts, supporting predictions from these species' different feeding ecologies. Furthermore, chimpanzees - but not bonobos - showed developmental improvements in spatial memory, indicating that bonobos exhibit cognitive paedomorphism (delays in developmental timing) in their spatial abilities relative to chimpanzees. Together, these results indicate that the development of spatial memory may differ even between closely related species. Moreover, changes in the spatial domain can emerge during nonhuman ape ontogeny, much like some changes seen in human children.

  13. DNA damage in cells exhibiting radiation-induced genomic instability

    DOE PAGESBeta

    Keszenman, Deborah J.; Kolodiuk, Lucia; Baulch, Janet E.

    2015-02-22

    Cells exhibiting radiation induced genomic instability exhibit varied spectra of genetic and chromosomal aberrations. Even so, oxidative stress remains a common theme in the initiation and/or perpetuation of this phenomenon. Isolated oxidatively modified bases, abasic sites, DNA single strand breaks and clustered DNA damage are induced in normal mammalian cultured cells and tissues due to endogenous reactive oxygen species generated during normal cellular metabolism in an aerobic environment. While sparse DNA damage may be easily repaired, clustered DNA damage may lead to persistent cytotoxic or mutagenic events that can lead to genomic instability. In this study, we tested the hypothesismore » that DNA damage signatures characterised by altered levels of endogenous, potentially mutagenic, types of DNA damage and chromosomal breakage are related to radiation-induced genomic instability and persistent oxidative stress phenotypes observed in the chromosomally unstable progeny of irradiated cells. The measurement of oxypurine, oxypyrimidine and abasic site endogenous DNA damage showed differences in non-double-strand breaks (DSB) clusters among the three of the four unstable clones evaluated as compared to genomically stable clones and the parental cell line. These three unstable clones also had increased levels of DSB clusters. The results of this study demonstrate that each unstable cell line has a unique spectrum of persistent damage and lead us to speculate that alterations in DNA damage signaling and repair may be related to the perpetuation of genomic instability.« less

  14. DNA damage in cells exhibiting radiation-induced genomic instability

    SciTech Connect

    Keszenman, Deborah J.; Kolodiuk, Lucia; Baulch, Janet E.

    2015-02-22

    Cells exhibiting radiation induced genomic instability exhibit varied spectra of genetic and chromosomal aberrations. Even so, oxidative stress remains a common theme in the initiation and/or perpetuation of this phenomenon. Isolated oxidatively modified bases, abasic sites, DNA single strand breaks and clustered DNA damage are induced in normal mammalian cultured cells and tissues due to endogenous reactive oxygen species generated during normal cellular metabolism in an aerobic environment. While sparse DNA damage may be easily repaired, clustered DNA damage may lead to persistent cytotoxic or mutagenic events that can lead to genomic instability. In this study, we tested the hypothesis that DNA damage signatures characterised by altered levels of endogenous, potentially mutagenic, types of DNA damage and chromosomal breakage are related to radiation-induced genomic instability and persistent oxidative stress phenotypes observed in the chromosomally unstable progeny of irradiated cells. The measurement of oxypurine, oxypyrimidine and abasic site endogenous DNA damage showed differences in non-double-strand breaks (DSB) clusters among the three of the four unstable clones evaluated as compared to genomically stable clones and the parental cell line. These three unstable clones also had increased levels of DSB clusters. The results of this study demonstrate that each unstable cell line has a unique spectrum of persistent damage and lead us to speculate that alterations in DNA damage signaling and repair may be related to the perpetuation of genomic instability.

  15. Liposomal nanoparticles encapsulating iloprost exhibit enhanced vasodilation in pulmonary arteries

    PubMed Central

    Jain, Pritesh P; Leber, Regina; Nagaraj, Chandran; Leitinger, Gerd; Lehofer, Bernhard; Olschewski, Horst; Olschewski, Andrea; Prassl, Ruth; Marsh, Leigh M

    2014-01-01

    Prostacyclin analogues are standard therapeutic options for vasoconstrictive diseases, including pulmonary hypertension and Raynaud’s phenomenon. Although effective, these treatment strategies are expensive and have several side effects. To improve drug efficiency, we tested liposomal nanoparticles as carrier systems. In this study, we synthesized liposomal nanoparticles tailored for the prostacyclin analogue iloprost and evaluated their pharmacologic efficacy on mouse intrapulmonary arteries, using a wire myograph. The use of cationic lipids, stearylamine, or 1,2-di-(9Z-octadecenoyl)-3-trimethylammonium-propane (DOTAP) in liposomes promoted iloprost encapsulation to at least 50%. The addition of cholesterol modestly reduced iloprost encapsulation. The liposomal nanoparticle formulations were tested for toxicity and pharmacologic efficacy in vivo and ex vivo, respectively. The liposomes did not affect the viability of human pulmonary artery smooth muscle cells. Compared with an equivalent concentration of free iloprost, four out of the six polymer-coated liposomal formulations exhibited significantly enhanced vasodilation of mouse pulmonary arteries. Iloprost that was encapsulated in liposomes containing the polymer polyethylene glycol exhibited concentration-dependent relaxation of arteries. Strikingly, half the concentration of iloprost in liposomes elicited similar pharmacologic efficacy as nonencapsulated iloprost. Cationic liposomes can encapsulate iloprost with high efficacy and can serve as potential iloprost carriers to improve its therapeutic efficacy. PMID:25045260

  16. Apolipophorin III from honeybees (Apis cerana) exhibits antibacterial activity.

    PubMed

    Kim, Bo Yeon; Jin, Byung Rae

    2015-04-01

    Apolipophorin III (apoLp-III) is involved in lipid transport and innate immunity in insects. In this study, an apoLp-III protein that exhibits antibacterial activity was identified in honeybees (Apis cerana). A. cerana apoLp-III cDNA encodes a 193 amino acid sequence that shares high identity with other members of the hymenopteran insect apoLp-III family. A. cerana apoLp-III is expressed constitutively in the fat body, epidermis, and venom gland and is detected as a 23-kDa protein. A. cerana apoLp-III expression is induced in the fat body after injection with Escherichia coli, Bacillus thuringiensis, or Beauveria bassiana. However, recombinant A. cerana apoLp-III (expressed in baculovirus-infected insect cells) binds directly to E. coli and B. thuringiensis but not to B. bassiana. Consistent with these findings, A. cerana apoLp-III exhibited antibacterial activity against both Gram-negative and Gram-positive bacteria. These results provide insight into the role of A. cerana apoLp-III during the innate immune response following bacterial infection.

  17. Peer review and competition in the Art Exhibition Game

    PubMed Central

    Goldstone, Robert L.; Helbing, Dirk

    2016-01-01

    To investigate the effect of competitive incentives under peer review, we designed a novel experimental setup called the Art Exhibition Game. We present experimental evidence of how competition introduces both positive and negative effects when creative artifacts are evaluated and selected by peer review. Competition proved to be a double-edged sword: on the one hand, it fosters innovation and product diversity, but on the other hand, it also leads to more unfair reviews and to a lower level of agreement between reviewers. Moreover, an external validation of the quality of peer reviews during the laboratory experiment, based on 23,627 online evaluations on Amazon Mechanical Turk, shows that competition does not significantly increase the level of creativity. Furthermore, the higher rejection rate under competitive conditions does not improve the average quality of published contributions, because more high-quality work is also rejected. Overall, our results could explain why many ground-breaking studies in science end up in lower-tier journals. Differences and similarities between the Art Exhibition Game and scholarly peer review are discussed and the implications for the design of new incentive systems for scientists are explained. PMID:27402744

  18. Chimpanzees and bonobos exhibit emotional responses to decision outcomes.

    PubMed

    Rosati, Alexandra G; Hare, Brian

    2013-01-01

    The interface between cognition, emotion, and motivation is thought to be of central importance in understanding complex cognitive functions such as decision-making and executive control in humans. Although nonhuman apes have complex repertoires of emotional expression, little is known about the role of affective processes in ape decision-making. To illuminate the evolutionary origins of human-like patterns of choice, we investigated decision-making in humans' closest phylogenetic relatives, chimpanzees (Pan troglodytes) and bonobos (Pan paniscus). In two studies, we examined these species' temporal and risk preferences, and assessed whether apes show emotional and motivational responses in decision-making contexts. We find that (1) chimpanzees are more patient and more risk-prone than are bonobos, (2) both species exhibit affective and motivational responses following the outcomes of their decisions, and (3) some emotional and motivational responses map onto species-level and individual-differences in decision-making. These results indicate that apes do exhibit emotional responses to decision-making, like humans. We explore the hypothesis that affective and motivational biases may underlie the psychological mechanisms supporting value-based preferences in these species.

  19. Nanocomposite thin films exhibiting high mechanical and optical flexibility

    NASA Astrophysics Data System (ADS)

    Druffel, Thad; Buazza, Omar; Lattis, Matt; Farmer, Scott

    2008-08-01

    Nanocomposites are created by doping host polymers with nanoparticles that typically have higher or lower refractive indices. The ability to tailor the mechanical and optical performance of these composites has led to their increased use in transparent materials. Nanocomposites maintain the elastic properties of the binding polymers and exhibit infinite refractive index tunability between the limits of the system. These unique properties provide distinct benefits for multilayer, thin-film optical filters. Because the nanoparticles are dispersed in a fluid or bound in a polymer matrix in use, toxicity risks that may be associated with raw particles are reduced. Using a stable dispersion of titanium dioxide nanoparticles and a UV curable monomer, we were able to design and produce several quarter-wave filters that demonstrate control of the height and width of the passband through adjustment of the organic/inorganic ratio and layer count. The volume loading of the metal oxides can be adjusted from zero to near the theoretical packing density of spheres, allowing refractive index to be controlled over a large range. Because metal oxide particles exhibit high UV absorption, these additives provide UV protection to the host polymer and the filter's substrate. Additionally, significant improvements in abrasion resistance are often observed in films loaded with nanoparticles at the concentrations of interest.

  20. Proteomic analysis of seminal fluid from men exhibiting oxidative stress

    PubMed Central

    2013-01-01

    Background Seminal plasma serves as a natural reservoir of antioxidants. It helps to remove excessive formation of reactive oxygen species (ROS) and consequently, reduce oxidative stress. Proteomic profiling of seminal plasma proteins is important to understand the molecular mechanisms underlying oxidative stress and sperm dysfunction in infertile men. Methods This prospective study consisted of 52 subjects: 32 infertile men and 20 healthy donors. Once semen and oxidative stress parameters were assessed (ROS, antioxidant concentration and DNA damage), the subjects were categorized into ROS positive (ROS+) or ROS negative (ROS-). Seminal plasma from each group was pooled and subjected to proteomics analysis. In-solution digestion and protein identification with liquid chromatography tandem mass spectrometry (LC-MS/MS), followed by bioinformatics analyses was used to identify and characterize potential biomarker proteins. Results A total of 14 proteins were identified in this analysis with 7 of these common and unique proteins were identified in both the ROS+ and ROS- groups through MASCOT and SEQUEST analyses, respectively. Prolactin-induced protein was found to be more abundantly present in men with increased levels of ROS. Gene ontology annotations showed extracellular distribution of proteins with a major role in antioxidative activity and regulatory processes. Conclusions We have identified proteins that help protect against oxidative stress and are uniquely present in the seminal plasma of the ROS- men. Men exhibiting high levels of ROS in their seminal ejaculate are likely to exhibit proteins that are either downregulated or oxidatively modified, and these could potentially contribute to male infertility. PMID:24004880

  1. Peer review and competition in the Art Exhibition Game.

    PubMed

    Balietti, Stefano; Goldstone, Robert L; Helbing, Dirk

    2016-07-26

    To investigate the effect of competitive incentives under peer review, we designed a novel experimental setup called the Art Exhibition Game. We present experimental evidence of how competition introduces both positive and negative effects when creative artifacts are evaluated and selected by peer review. Competition proved to be a double-edged sword: on the one hand, it fosters innovation and product diversity, but on the other hand, it also leads to more unfair reviews and to a lower level of agreement between reviewers. Moreover, an external validation of the quality of peer reviews during the laboratory experiment, based on 23,627 online evaluations on Amazon Mechanical Turk, shows that competition does not significantly increase the level of creativity. Furthermore, the higher rejection rate under competitive conditions does not improve the average quality of published contributions, because more high-quality work is also rejected. Overall, our results could explain why many ground-breaking studies in science end up in lower-tier journals. Differences and similarities between the Art Exhibition Game and scholarly peer review are discussed and the implications for the design of new incentive systems for scientists are explained. PMID:27402744

  2. Parallax Park - A Bilingual, Outdoor, Interactive Family Exhibit

    NASA Astrophysics Data System (ADS)

    Benedict, G. F.; Parallax Park Design Team

    2005-10-01

    Parallax Park will be an outdoor, interactive family exhibit for McDonald Observatory. It will bring the quest for extra-solar planets and the astronomical distance scale down to Earth for the more than 100,000 visitors who come to the Observatory each year. The Park will teach visitors the basic principles of astrometry. It will concentrate on distance determination and extrasolar planet detection, and emphasize the advantages of space-based astrometry measurements. Visitors will experience parallax by traveling a path around a representation of the Sun, mimicking Earth's orbit, while viewing representations of stars at various distances and in various directions. Concepts include parallax, Cepheids as standard candles, the inverse square law, and motion around a center of mass. The exhibit includes interactive components suitable for children's use. Text labels and printed guides, in English and Spanish, will explain how to use the Park to explore the uses of astrometry in modern science. In addition auxiliary educational materials that align with the National Science Education Standards will be produced.

  3. In vitro hydrogen production by glucose dehydrogenase and hydrogenase

    SciTech Connect

    Woodward, J.; Mattingly, S.M.; Danson, M.

    1996-07-01

    A new in vitro enzymatic pathway for the generation of molecular hydrogen from glucose has been demonstrated. The reaction is based on the oxidation of glucose by Thermoplasma acidophilum glucose dehydrogenase with the concomitant oxidation of NADPH by Pyrococcus furiosus hydrogenase. Stoichiometric yields of hydrogen were produced from glucose with the continuous recycling of cofactor. This simple system may provide a method for the biological production of hydrogen from renewable sources. In addition, the other product of this reaction, gluconic acid, is a high-value chemical commodity. 23 refs., 5 figs.

  4. In vitro hydrogen production by glucose dehydrogenase and hydrogenase

    SciTech Connect

    Woodward, J.

    1996-10-01

    A new in vitro enzymatic pathway for the generation of molecular hydrogen from glucose has been demonstrated. The reaction is based upon the oxidation of glucose by Thermoplasma acidophilum glucose dehydrogenase with the concomitant oxidation of NADPH by Pyrococcus furiosus hydrogenase. Stoichiometric yields of hydrogen were produced from glucose with continuous cofactor recycle. This simple system may provide a method for the biological production of hydrogen from renewable sources. In addition, the other product of this reaction, gluconic acid, is a high-value commodity chemical.

  5. NMR studies on mechanism of isomerisation of fructose 6-phosphate to glucose 6-phosphate catalysed by phosphoglucose isomerase from Thermococcus kodakarensis.

    PubMed

    Abbas, Shahzada Nadeem; Mok, Kenneth Hun; Rashid, Naeem; Xie, Yongjing; Ruether, Manuel; O'Brien, John; Akhtar, Muhammad

    2016-06-01

    The fate of hydrogen atoms at C-2 of glucose 6-phosphate (G6P) and C-1 of fructose 6-phosphate (F6P) was studied in the reaction catalysed by phosphoglucose isomerase from Thermococcus kodakarensis (TkPGI) through 1D and 2D NMR methods. When the reaction was performed in (2)H2O the hydrogen atoms in the aforementioned positions were exchanged with deuterons indicating that the isomerization occurred by a cis-enediol intermediate involving C-1 pro-R hydrogen of F6P. These features are similar to those described for phosphoglucose isomerases from rabbit muscle and Pyrococcus furiosus.

  6. Combining biomass wet disk milling and endoglucanase/β-glucosidase hydrolysis for the production of cellulose nanocrystals.

    PubMed

    Teixeira, Ricardo Sposina Sobral; da Silva, Ayla Sant'Ana; Jang, Jae-Hyuk; Kim, Han-Woo; Ishikawa, Kazuhiko; Endo, Takashi; Lee, Seung-Hwan; Bon, Elba P S

    2015-09-01

    Cellulose nanocrystals (CNCs), a biomaterial with high added value, were obtained from pure cellulose, Eucalyptus holocellulose, unbleached Kraft pulp, and sugarcane bagasse, by fibrillating these biomass substrates using wet disk milling (WDM) followed by enzymatic hydrolysis using endoglucanase/β-glucosidase. The hydrolysis experiments were conducted using the commercial enzyme OptimashBG or a blend of Pyrococcus horikoshii endoglucanase and Pyrococcus furiosus β-glucosidase. The fibrillated materials and CNCs were analyzed by X-ray diffraction, atomic force microscopy, scanning electron microscopy, and the specific surface area (SSA) was measured. WDM resulted in the formation of long and twisted microfibers of 1000-5000 nm in length and 4-35 nm in diameter, which were hydrolyzed into shorter and straighter CNCs of 500-1500 nm in length and 4-12 nm in diameter, with high cellulose crystallinity. Therefore, the CNC's aspect ratio was successfully adjusted by endoglucanases under mild reaction conditions, relative to the reported acidic hydrolysis method.

  7. Use of cellobiohydrolase-free cellulase blends for the hydrolysis of microcrystalline cellulose and sugarcane bagasse pretreated by either ball milling or ionic liquid [Emim][Ac].

    PubMed

    Teixeira, Ricardo Sposina Sobral; da Silva, Ayla Sant'Ana; Kim, Han-Woo; Ishikawa, Kazuhiko; Endo, Takashi; Lee, Seung-Hwan; Bon, Elba P S

    2013-12-01

    This study investigated the requirement of cellobiohydrolases (CBH) for saccharification of microcrystalline cellulose and sugarcane bagasse pretreated either by ball milling (BM) or by ionic liquid (IL) [Emim][Ac]. Hydrolysis was done using CBH-free blends of Pyrococcus horikoshii endoglucanase (EG) plus Pyrococcus furiosus β-glucosidase (EGPh/BGPf) or Optimash™ BG while Acremonium Cellulase was used as control. IL-pretreated substrates were hydrolyzed more effectively by CBH-free enzymes than were the BM-pretreated substrates. IL-treatment decreased the crystallinity and increased the specific surface area (SSA), whereas BM-treatment decreased the crystallinity without increasing the SSA. The hydrolysis of IL-treated cellulose by EGPh/BGPf showed a saccharification rate of 3.92 g/Lh and a glucose yield of 81% within 9h. These results indicate the efficiency of CBH-free enzymes for the hydrolysis of IL-treated substrates.

  8. Plants at high altitude exhibit higher component of alternative respiration.

    PubMed

    Kumar, Narinder; Vyas, Dhiraj; Kumar, Sanjay

    2007-01-01

    Total respiration, capacities of cytochrome (CytR) and alternative respiration (AR) were studied in two varieties of barley (Horedum vulgare) and wheat (Triticum aestivum) each and one variety of pea (Pisum sativum) at low (Palampur; 1300 m) and high altitudes (Kibber; 4200 m). Similar studies were carried out in naturally growing Rumex nepalensis and Trifoilum repenses at Palampur, Palchan (2250 m) and Marhi (3250 m). All the plants species exhibited lower CytR but significantly higher AR capacity at high altitude (HA) (72-1117% higher) as compared to those at low altitude (LA). Glycolytic product, pyruvate and tricarboxylic acid cycle intermediate, citrate increased with increase in altitude. While the role of these metabolites in relation to HA biology is discussed, significantly higher AR at HA is proposed to be an adaptive mechanism against the metabolic perturbations wherein it might act to lower reactive oxygen species and also provides metabolic homeostasis to plants under the environment of HA.

  9. Achiral flexible liquid crystal trimers exhibiting chiral conglomerates.

    PubMed

    Sasaki, Haruna; Takanishi, Yoichi; Yamamoto, Jun; Yoshizawa, Atsushi

    2016-04-14

    Chiral conglomerates of domains with opposite handedness have attracted much attention from researchers. We prepared a homologous series of achiral liquid crystal trimers in which two phenylpyrimidine units and one biphenyl unit were connected via flexible methylene spacers. We investigated their phase transition behaviour. Some trimers possessing odd-numbered spacers were found to exhibit a nematic phase and a dark chiral conglomerate phase possessing a layered structure. The chiral characteristics were confirmed by uncrossing the polarizers in opposite directions. The layer spacing detected using X-ray diffraction was about 80% of the molecular length. The structure-property relations indicate that intermolecular interactions cause a conformational change in the trimers possessing flexible odd-numbered methylene spacers to form helical conformers with axial chirality, which might induce chiral segregation and layer deformation to drive the chiral conglomerates.

  10. A novel cyanobacterium exhibiting an elevated tolerance for iron.

    PubMed

    Brown, Igor I; Mummey, Daniel; Cooksey, Keith E

    2005-05-01

    Studies directed at cyanobacteria inhabiting iron-depositing hot springs may provide insights into the role of both ancient and contemporary cyanobacteria mediated iron transformations. Here we phylogenetically, morphologically and physiologically characterize a novel cyanobacterium isolated from an iron-depositing hot spring. Phylogenetic analyses indicate that the bacterium is a representative of a new genus, exhibiting a maximum 95.2% homology to database sequences. The isolate is a unicellular cyanobacterium with bladder-like cells typically packed as duplexes, or in extracellular polymeric substance covered clumps and small chains without the ability to produce baeocystes. No growth without added combined nitrogen occurred. While requiring relatively large amounts of iron for growth (>40microM), the isolate was shown to facilitate removal of iron from culture media. These results suggest that the isolate may be an important component of an iron-depositing microbial community. The name "Chroogloeocystis siderophila" for this cyanobacterium is proposed. PMID:16329916

  11. [A case of Williams syndrome who exhibited fetishism].

    PubMed

    Noguchi, Masayuki; Kato, Satoshi

    2004-01-01

    Williams syndrome is a rare congenital disease in which the etiological locus is a micro-deletion in chromosome 7. Here, we describe the case of a 22-year-old male who was diagnosed with Williams syndrome at the age of 3 years. As a child, the patient exhibited patterns of behavior characteristic of this syndrome including hyperactivity, attention deficit, and over-friendliness. He also showed persistent interest in construction vehicles, playgrounds, and gloves. He became interested in gloves after watching a television program in which the heroine fought her enemies while wearing gloves. Watching pornographic movies allowed him to attach strong sexual significance to gloves when he was 19 years old. Since that time, he has assaulted women wearing gloves four times to rob them of the gloves. The current paper discusses both the role of the cognitive profile unique to Williams syndrome and that of environmental factors in the development of fetishism in this case. PMID:15669216

  12. Ultrasensitive dual phosphorylation dephosphorylation cycle kinetics exhibits canonical competition behavior

    NASA Astrophysics Data System (ADS)

    Huang, Qingdao; Qian, Hong

    2009-09-01

    We establish a mathematical model for a cellular biochemical signaling module in terms of a planar differential equation system. The signaling process is carried out by two phosphorylation-dephosphorylation reaction steps that share common kinase and phosphatase with saturated enzyme kinetics. The pair of equations is particularly simple in the present mathematical formulation, but they are singular. A complete mathematical analysis is developed based on an elementary perturbation theory. The dynamics exhibits the canonical competition behavior in addition to bistability. Although widely understood in ecological context, we are not aware of a full range of biochemical competition in a simple signaling network. The competition dynamics has broad implications to cellular processes such as cell differentiation and cancer immunoediting. The concepts of homogeneous and heterogeneous multisite phosphorylation are introduced and their corresponding dynamics are compared: there is no bistability in a heterogeneous dual phosphorylation system. A stochastic interpretation is also provided that further gives intuitive understanding of the bistable behavior inside the cells.

  13. The Pahrump Valley Museum Yucca Mountain History Exhibit - 12389

    SciTech Connect

    Voegele, Michael; McCracken, Robert; Herrera, Troy

    2012-07-01

    As part of its management of the Yucca Mountain project, the Department of Energy maintained several information centers to provide public access to information about the status of the Yucca Mountain project. Those information centers contained numerous displays, historical information, and served as the location for the Department's outreach activities. As the Department of Energy dealt with reduced budgets in 2009 following the Obama Administration's intent to terminate the program, it shut down its information centers. Nye County considered it important to maintain a public information center where people would be able to find information about what was happening with the Yucca Mountain project. Initially the Nye County assumed responsibility for the information center in Pahrump; eventually the County made a decision to move that information center into an expansion of the existing Pahrump Valley Museum. Nye County undertook an effort to update the information about the Yucca Mountain project and modernize the displays. A parallel effort to create a source of historical information where people could find out about the Yucca Mountain project was undertaken. To accompany the Yucca Mountain exhibits in the Pahrump Valley Museum, Nye County also sponsored a series of interviews to document, through oral histories, as much information about the Yucca Mountain project as could be found in these interviews. The paper presents an overview of the Yucca Mountain exhibits in the Pahrump Valley Museum, and the accompanying oral histories. An important conclusion that can be drawn from the interviews is that construction of a repository in Nevada should have been conceptualized as but the first step in transforming the economy of central Nevada by turning part of the Nevada National Security Site and adjoining area into a world-class energy production and energy research center. (authors)

  14. Wave velocity dispersion and attenuation in media exhibiting internal oscillations

    NASA Astrophysics Data System (ADS)

    Frehner, Marcel; Steeb, Holger; Schmalholz, Stefan M.

    2010-05-01

    Understanding the dynamical and acoustical behavior of porous and heterogeneous rocks is of great importance in geophysics, e.g. earthquakes, and for various seismic engineering applications, e.g. hydrocarbon exploration. Within a heterogeneous medium oscillations with a characteristic resonance frequency, depending on the mass and internal length of the heterogeneity, can occur. When excited, heterogeneities can self-oscillate with their natural frequency. Another example of internal oscillations is the dynamical behavior of non-wetting fluid blobs or fluid patches in residually saturated pore spaces. Surface tension forces or capillary forces act as the restoring force that drives the oscillation. Whatever mechanism is involved, an oscillatory phenomena within a heterogeneous medium will have an effect on acoustic or seismic waves propagating through such a medium, i.e. wave velocity dispersion and frequency-dependent attenuation. We present two models for media exhibiting internal oscillations and discuss the frequency-dependent wave propagation mechanism. Both models give similar results: (1) The low-frequency (i.e. quasi-static) limit for the phase velocity is identical with the Gassmann-Wood limit and the high-frequency limit is larger than this value and (2) Around the resonance frequency a very strong phase velocity change and the largest attenuation occurs. (1) Model for a homogeneous medium exhibiting internal oscillations We present a continuum model for an acoustic medium exhibiting internal damped oscillations. The obvious application of this model is water containing oscillating gas bubbles, providing the material and model parameters for this study. Two physically based momentum interaction terms between the two inherent constituents are used: (1) A purely elastic term of oscillatory nature that scales with the volume of the bubbles and (2) A viscous term that scales with the specific surface of the bubble. The model is capable of taking into account

  15. Ultralyophobic oxidized aluminum surfaces exhibiting negligible contact angle hysteresis.

    PubMed

    Hozumi, Atsushi; McCarthy, Thomas J

    2010-02-16

    Ultralyophobic oxidized aluminum surfaces exhibiting negligible contact angle hysteresis for probe liquids were prepared by chemical vapor deposition (CVD) of bis((tridecafluoro-1,1,2,2,-tetrahydrooctyl)-dimethylsiloxy)methylsilane (CF(3)(CF(2))(5)CH(2)CH(2)Si(CH(3))(2)O)(2)SiCH(3)H, (R(F)Si(Me)(2)O)(2)SiMeH). Oxidized aluminum surfaces were prepared by photooxidation/cleaning of sputter-coated aluminum on silicon wafers (Si/Al(Al(2)(O(3)))) using oxygen plasma. X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM) confirmed that this facile CVD method produces a monolayer with a thickness of 1.1 nm on the Si/Al(Al(2)(O(3))) surface without a discernible change in surface morphology. After monolayer deposition, the hydrophilic Si/Al(Al(2)(O(3))) surface became both hydrophobic and oleophobic and exhibited essentially no contact angle hysteresis for water and n-hexadecane (advancing/receding contact angles (theta(A)/theta(R)) = 110 degrees/109 degrees and 52 degrees/50 degrees, respectively). Droplets move very easily on this surface and roll off of slightly tilted surfaces, independently of the contact angle (which is a practical definition of ultralyophobic). A conventional fluoroalkylsilane monolayer was also prepared from 1H,1H,2H,2H-perfluorodecyltrimethoxysilane (CF(3)(CF(2))(7)CH(2)CH(2)Si(OCH(3))(3), R(F)Si(OMe)(3)) for comparison. The theta(A)/theta(R) values for water and n-hexadecane are 121 degrees/106 degrees and 76 degrees/71 degrees, respectively. The larger hysteresis values indicate the "pinning" of probe liquids, even though advancing contact angles are larger than those of the (R(F)Si(Me)(2)O)(2)SiMeH-derived monolayers. The (R(F)Si(Me)(2)O)(2)SiMeH-derived monolayers have excellent hydrolytic stability in water. We propose that the (R(F)Si(Me)(2)O)(2)SiMeH-derived monolayers are flexible and liquidlike and that drops in contact with these surfaces experience very low energy barriers between metastable states, leading to the

  16. How Can Museum Exhibits Enhance Earthquake and Tsunami Hazard Resiliency?

    NASA Astrophysics Data System (ADS)

    Olds, S. E.

    2015-12-01

    Creating a natural disaster-ready community requires interoperating scientific, technical, and social systems. In addition to the technical elements that need to be in place, communities and individuals need to be prepared to react when a natural hazard event occurs. Natural hazard awareness and preparedness training and education often takes place through informal learning at science centers and formal k-12 education programs as well as through awareness raising via strategically placed informational tsunami warning signs and placards. Museums and science centers are influential in raising science literacy within a community, however can science centers enhance earthquake and tsunami resiliency by providing hazard science content and preparedness exhibits? Museum docents and informal educators are uniquely situated within the community. They are transmitters and translators of science information to broad audiences. Through interaction with the public, docents are well positioned to be informants of the knowledge beliefs, and feelings of science center visitors. They themselves are life-long learners, both constantly learning from the museum content around them and sharing this content with visitors. They are also members of a community where they live. In-depth interviews with museum informal educators and docents were conducted at a science center in coastal Pacific Northwest. This region has a potential to be struck by a great 9+ Mw earthquake and subsequent tsunami. During the interviews, docents described how they applied learning from natural hazard exhibits at a science visitor center to their daily lives. During the individual interviews, the museum docents described their awareness (knowledge, attitudes, and behaviors) of natural hazards where they live and work, the feelings evoked as they learned about their hazard vulnerability, the extent to which they applied this learning and awareness to their lives, such as creating an evacuation plan, whether

  17. Ultralyophobic oxidized aluminum surfaces exhibiting negligible contact angle hysteresis.

    PubMed

    Hozumi, Atsushi; McCarthy, Thomas J

    2010-02-16

    Ultralyophobic oxidized aluminum surfaces exhibiting negligible contact angle hysteresis for probe liquids were prepared by chemical vapor deposition (CVD) of bis((tridecafluoro-1,1,2,2,-tetrahydrooctyl)-dimethylsiloxy)methylsilane (CF(3)(CF(2))(5)CH(2)CH(2)Si(CH(3))(2)O)(2)SiCH(3)H, (R(F)Si(Me)(2)O)(2)SiMeH). Oxidized aluminum surfaces were prepared by photooxidation/cleaning of sputter-coated aluminum on silicon wafers (Si/Al(Al(2)(O(3)))) using oxygen plasma. X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM) confirmed that this facile CVD method produces a monolayer with a thickness of 1.1 nm on the Si/Al(Al(2)(O(3))) surface without a discernible change in surface morphology. After monolayer deposition, the hydrophilic Si/Al(Al(2)(O(3))) surface became both hydrophobic and oleophobic and exhibited essentially no contact angle hysteresis for water and n-hexadecane (advancing/receding contact angles (theta(A)/theta(R)) = 110 degrees/109 degrees and 52 degrees/50 degrees, respectively). Droplets move very easily on this surface and roll off of slightly tilted surfaces, independently of the contact angle (which is a practical definition of ultralyophobic). A conventional fluoroalkylsilane monolayer was also prepared from 1H,1H,2H,2H-perfluorodecyltrimethoxysilane (CF(3)(CF(2))(7)CH(2)CH(2)Si(OCH(3))(3), R(F)Si(OMe)(3)) for comparison. The theta(A)/theta(R) values for water and n-hexadecane are 121 degrees/106 degrees and 76 degrees/71 degrees, respectively. The larger hysteresis values indicate the "pinning" of probe liquids, even though advancing contact angles are larger than those of the (R(F)Si(Me)(2)O)(2)SiMeH-derived monolayers. The (R(F)Si(Me)(2)O)(2)SiMeH-derived monolayers have excellent hydrolytic stability in water. We propose that the (R(F)Si(Me)(2)O)(2)SiMeH-derived monolayers are flexible and liquidlike and that drops in contact with these surfaces experience very low energy barriers between metastable states, leading to the

  18. Isolation and characterization of novel bacterial strains exhibiting ligninolytic potential

    PubMed Central

    2011-01-01

    Background To expand on the range of products which can be obtained from lignocellulosic biomass, the lignin component should be utilized as feedstock for value-added chemicals such as substituted aromatics, instead of being incinerated for heat and energy. Enzymes could provide an effective means for lignin depolymerization into products of interest. In this study, soil bacteria were isolated by enrichment on Kraft lignin and evaluated for their ligninolytic potential as a source of novel enzymes for waste lignin valorization. Results Based on 16S rRNA gene sequencing and phenotypic characterization, the organisms were identified as Pandoraea norimbergensis LD001, Pseudomonas sp LD002 and Bacillus sp LD003. The ligninolytic capability of each of these isolates was assessed by growth on high-molecular weight and low-molecular weight lignin fractions, utilization of lignin-associated aromatic monomers and degradation of ligninolytic indicator dyes. Pandoraea norimbergensis LD001 and Pseudomonas sp. LD002 exhibited best growth on lignin fractions, but limited dye-decolourizing capacity. Bacillus sp. LD003, however, showed least efficient growth on lignin fractions but extensive dye-decolourizing capacity, with a particular preference for the recalcitrant phenothiazine dye class (Azure B, Methylene Blue and Toluidene Blue O). Conclusions Bacillus sp. LD003 was selected as a promising source of novel types of ligninolytic enzymes. Our observations suggested that lignin mineralization and depolymerization are separate events which place additional challenges on the screening of ligninolytic microorganisms for specific ligninolytic enzymes. PMID:21995752

  19. Glucose biosensor based on multisegment nanowires exhibiting reversible magnetic control.

    PubMed

    Gerola, Gislaine P; Takahashi, Giovanna S; Perez, Geraldo G; Recco, Lucas C; Pedrosa, Valber A

    2014-11-01

    We describe the amperometric detection of glucose using oriented nanowires with magnetic switching of the bioelectrochemical process. The fabrication process of the nanowires was prepared through controlled nucleation and growth during a stepwise electrochemical deposition, and it was characterized using scanning electron microscopy. Cyclic voltammetry and amperometry were used to study the magnetoswitchable property; this control was accomplished by changing the surface orientation of nanowires. Under the optimal condition, the amperometric response was also linear up to a glucose concentration of 0.1-16.0 mmol L(-1) with a sensitivity of 81 μA mM(-1). The detection limit was estimated for 4.8×10(-8) mol L(-1), defined from a signal/noise ratio of 3. It also exhibits good reproducibility and high selectivity with insignificant interference from ascorbic acid, acetoaminophen, and uric acid. The resulting biosensor was applied to detect the blood sugar in human serum samples without any pretreatment, and the results were comparatively in agreement with the clinical assay. PMID:25127595

  20. Norepinephrine transporter heterozygous knockout mice exhibit altered transport and behavior.

    PubMed

    Fentress, H M; Klar, R; Krueger, J J; Sabb, T; Redmon, S N; Wallace, N M; Shirey-Rice, J K; Hahn, M K

    2013-11-01

    The norepinephrine (NE) transporter (NET) regulates synaptic NE availability for noradrenergic signaling in the brain and sympathetic nervous system. Although genetic variation leading to a loss of NET expression has been implicated in psychiatric and cardiovascular disorders, complete NET deficiency has not been found in people, limiting the utility of NET knockout mice as a model for genetically driven NET dysfunction. Here, we investigate NET expression in NET heterozygous knockout male mice (NET(+/-) ), demonstrating that they display an approximately 50% reduction in NET protein levels. Surprisingly, these mice display no significant deficit in NET activity assessed in hippocampal and cortical synaptosomes. We found that this compensation in NET activity was due to enhanced activity of surface-resident transporters, as opposed to surface recruitment of NET protein or compensation through other transport mechanisms, including serotonin, dopamine or organic cation transporters. We hypothesize that loss of NET protein in the NET(+/-) mouse establishes an activated state of existing surface NET proteins. The NET(+/-) mice exhibit increased anxiety in the open field and light-dark box and display deficits in reversal learning in the Morris water maze. These data suggest that recovery of near basal activity in NET(+/-) mice appears to be insufficient to limit anxiety responses or support cognitive performance that might involve noradrenergic neurotransmission. The NET(+/-) mice represent a unique model to study the loss and resultant compensatory changes in NET that may be relevant to behavior and physiology in human NET deficiency disorders.

  1. Polymeric surfaces exhibiting photocatalytic activity and controlled anisotropic wettability

    NASA Astrophysics Data System (ADS)

    Anastasiadis, Spiros H.; Frysali, Melani A.; Papoutsakis, Lampros; Kenanakis, George; Stratakis, Emmanuel; Vamvakaki, Maria; Mountrichas, Grigoris; Pispas, Stergios

    2015-03-01

    In this work we focus on surfaces, which exhibit controlled, switchable wettability in response to one or more external stimuli as well as photocatalytic activity. For this we are inspired from nature to produce surfaces with a dual-scale hierarchical roughness and combine them with the appropriate inorganic and/or polymer coating. The combination of the hierarchical surface with a ZnO coating and a pH- or temperature-responsive polymer results in efficient photo-active properties as well as reversible superhydrophobic / superhydrophilic surfaces. Furthermore, we fabricate surfaces with unidirectional wettability variation. Overall, such complex surfaces require advanced design, combining hierarchically structured surfaces with suitable polymeric materials. Acknowledgment: This research was partially supported by the European Union (European Social Fund, ESF) and Greek national funds through the ``ARISTEIA II'' Action (SMART-SURF) of the Operational Programme ``Education and Lifelong Learning,'' NSRF 2007-2013, via the General Secretariat for Research & Technology, Ministry of Education and Religious Affairs, Greece.

  2. Method for making glass-ceramic articles exhibiting high frangibility

    DOEpatents

    Beall, George H.; Brydges, III., William T.; Ference, Joseph; Kozlowski, Theodore R.

    1976-02-03

    This invention is concerned with glass-ceramic articles having compositions within a very narrowly-delimited area of the MgO-Al.sub.2 O.sub.3 -B.sub.2 O.sub.3 -SiO.sub.2 field and having alpha-quartz and sapphirine as the principal crystal phases, resulting from nucleation through a combination of TiO.sub.2 and ZrO.sub.2. Upon contacting such articles with lithium ions at an elevated temperature, said lithium ions will replace magnesium ions on a two Li.sup.+-for-one Mg.sup..sup.+2 basis within the crystal structures, thereby providing a unitary glass-ceramic article having an integral surface layer wherein the principal crystal phase is a lithium-stuffed beta-quartz solid solution. That transformation of crystal phases results in compressive stresses being set up within the surface layer as the articles are cooled. Through the careful control of composition, crystallization treatment, and the parameters of the replacement reaction in the crystal structures, a tremendous degree of stored elastic energy can be developed within the articles such that they will demonstrate frangibility when fractured but will not exhibit undesirable spontaneous breakage and/or spalling.

  3. Metallic Zinc Exhibits Optimal Biocompatibility for Bioabsorbable Endovascular Stents

    PubMed Central

    Bowen, Patrick K.; Guillory, Roger J.; Shearier, Emily R.; Seitz, Jan-Marten; Drelich, Jaroslaw; Bocks, Martin; Zhao, Feng; Goldman, Jeremy

    2015-01-01

    Although corrosion resistant bare metal stents are considered generally effective, their permanent presence in a diseased artery is an increasingly recognized limitation due to the potential for long-term complications. We previously reported that metallic zinc exhibited an ideal biocorrosion rate within murine aortas, thus raising the possibility of zinc as a candidate base material for endovascular stenting applications. This study was undertaken to further assess the arterial biocompatibility of metallic zinc. Metallic zinc wires were punctured and advanced into the rat abdominal aorta lumen for up to 6.5 months. This study demonstrated that metallic zinc did not provoke responses that often contribute to restenosis. Low cell densities and neointimal tissue thickness, along with tissue regeneration within the corroding implant, point to optimal biocompatibility of corroding zinc. Furthermore, the lack of progression in neointimal tissue thickness over 6.5 months or the presence of smooth muscle cells near the zinc implant suggest that the products of zinc corrosion may suppress the activities of inflammatory and smooth muscle cells. PMID:26249616

  4. Pimecrolimus micelle exhibits excellent therapeutic effect for Keratoconjunctivitis Sicca.

    PubMed

    Yingfang, Fan; Zhuang, Bo; Wang, Cheng; Xu, Xuelian; Xu, Wei; Lv, Zhihua

    2016-04-01

    Poor corneal penetration and short residence time on the ocular surface are two major bottlenecks for conventional ophthalmic formulations. To overcome the foregoing dilemmas, we prepared two novel formulations of pimecrolimus nanomicelles (PNM) with particle size of 37.85 ± 1.21 nm and thermosensitive hydrogel (PTH) for treating Keratoconjunctivitis Sicca (KCS). PNM were investigated by transmission electron microscopy (TEM), Malvern laser particle size analyzer, X-ray diffraction (XRD) system, and the content of drug in PNM was measured by high-performance liquid chromatography (HPLC). The drug loading and encapsulation efficiency reached to 7.57% ± 0.10% and 97.9% ± 1.26%, respectively. PTH displayed special gel-sol transition behavior with temperature increasing from 4 °C to 37 °C. The in vitro release profile demonstrated that PNM and PTH exhibited sustained-release behavior compared with free pimecrolimus oil-based eye drop (FPO). In addition, we established a mouse model of KCS induced by benzalkonium chloride to evaluate the therapeutic outcome of different pimecrolimus formulations. The production of tear, fluorescein staining scores and histopathologic examinations of the cornea were assessed in detail. The results confirmed that PNM had the best therapeutic effect among all formulations based on its higher drug encapsulation capability, favourable permeability and sustained release. All these indicated that PNM could serve as a potent ophthalmologic agent for KCS. PMID:26731192

  5. Bonobos and chimpanzees exhibit human-like framing effects.

    PubMed

    Krupenye, Christopher; Rosati, Alexandra G; Hare, Brian

    2015-02-01

    Humans exhibit framing effects when making choices, appraising decisions involving losses differently from those involving gains. To directly test for the evolutionary origin of this bias, we examined decision-making in humans' closest living relatives: bonobos (Pan paniscus) and chimpanzees (Pan troglodytes). We presented the largest sample of non-humans to date (n = 40) with a simple task requiring minimal experience. Apes made choices between a 'framed' option that provided preferred food, and an alternative option that provided a constant amount of intermediately preferred food. In the gain condition, apes experienced a positive 'gain' event in which the framed option was initially presented as one piece of food but sometimes was augmented to two. In the loss condition, apes experienced a negative 'loss' event in which they initially saw two pieces but sometimes received only one. Both conditions provided equal pay-offs, but apes chose the framed option more often in the positive 'gain' frame. Moreover, male apes were more susceptible to framing than were females. These results suggest that some human economic biases are shared through common descent with other apes and highlight the importance of comparative work in understanding the origins of individual differences in human choice. PMID:25672997

  6. Growth states of catalytic reaction networks exhibiting energy metabolism

    NASA Astrophysics Data System (ADS)

    Kondo, Yohei; Kaneko, Kunihiko

    2011-07-01

    All cells derive nutrition by absorbing some chemical and energy resources from the environment; these resources are used by the cells to reproduce the chemicals within them, which in turn leads to an increase in their volume. In this study we introduce a protocell model exhibiting catalytic reaction dynamics, energy metabolism, and cell growth. Results of extensive simulations of this model show the existence of four phases with regard to the rates of both the influx of resources and cell growth. These phases include an active phase with high influx and high growth rates, an inefficient phase with high influx but low growth rates, a quasistatic phase with low influx and low growth rates, and a death phase with negative growth rate. A mean field model well explains the transition among these phases as bifurcations. The statistical distribution of the active phase is characterized by a power law, and that of the inefficient phase is characterized by a nearly equilibrium distribution. We also discuss the relevance of the results of this study to distinct states in the existing cells.

  7. Semiparametric energy-based models of systems exhibiting criticality

    NASA Astrophysics Data System (ADS)

    Humplik, Jan; Tkacik, Gasper

    Over the last decade, several empirical studies have found evidence that many biological and natural systems exhibit critical fluctuations analogous to those observed during second-order phase transitions in equilibrium systems. In many cases, these fluctuations were shown to be equivalent to a thermodynamic version of Zipf's law-if the system is sufficiently large, then a log-log plot of the probability of a state vs. its rank yields a straight line with slope - 1 . Because the origin of critical fluctuations cannot be traced to a unique mechanism, it is important that data-driven phenomenological models of natural systems are flexible enough so as to easily capture any kind of criticality. Here we study a class of models with exactly this property. This class consists of energy-based models in which the exponential Boltzmann factor is replaced by an arbitrary nonlinear function. We demonstrate the usefulness of our method by modeling the spiking activity of a population of retinal neurons, and the distribution of light intensities in small patches of natural images. In light of recent work on models with hidden variables, the proposed method can separate interactions induced by an unknown fluctuating environment from interactions intrinsic to the system.

  8. Metallic zinc exhibits optimal biocompatibility for bioabsorbable endovascular stents.

    PubMed

    Bowen, Patrick K; Guillory, Roger J; Shearier, Emily R; Seitz, Jan-Marten; Drelich, Jaroslaw; Bocks, Martin; Zhao, Feng; Goldman, Jeremy

    2015-11-01

    Although corrosion resistant bare metal stents are considered generally effective, their permanent presence in a diseased artery is an increasingly recognized limitation due to the potential for long-term complications. We previously reported that metallic zinc exhibited an ideal biocorrosion rate within murine aortas, thus raising the possibility of zinc as a candidate base material for endovascular stenting applications. This study was undertaken to further assess the arterial biocompatibility of metallic zinc. Metallic zinc wires were punctured and advanced into the rat abdominal aorta lumen for up to 6.5months. This study demonstrated that metallic zinc did not provoke responses that often contribute to restenosis. Low cell densities and neointimal tissue thickness, along with tissue regeneration within the corroding implant, point to optimal biocompatibility of corroding zinc. Furthermore, the lack of progression in neointimal tissue thickness over 6.5months or the presence of smooth muscle cells near the zinc implant suggest that the products of zinc corrosion may suppress the activities of inflammatory and smooth muscle cells.

  9. Growth states of catalytic reaction networks exhibiting energy metabolism.

    PubMed

    Kondo, Yohei; Kaneko, Kunihiko

    2011-07-01

    All cells derive nutrition by absorbing some chemical and energy resources from the environment; these resources are used by the cells to reproduce the chemicals within them, which in turn leads to an increase in their volume. In this study we introduce a protocell model exhibiting catalytic reaction dynamics, energy metabolism, and cell growth. Results of extensive simulations of this model show the existence of four phases with regard to the rates of both the influx of resources and cell growth. These phases include an active phase with high influx and high growth rates, an inefficient phase with high influx but low growth rates, a quasistatic phase with low influx and low growth rates, and a death phase with negative growth rate. A mean field model well explains the transition among these phases as bifurcations. The statistical distribution of the active phase is characterized by a power law, and that of the inefficient phase is characterized by a nearly equilibrium distribution. We also discuss the relevance of the results of this study to distinct states in the existing cells. PMID:21867233

  10. Glucose biosensor based on multisegment nanowires exhibiting reversible magnetic control.

    PubMed

    Gerola, Gislaine P; Takahashi, Giovanna S; Perez, Geraldo G; Recco, Lucas C; Pedrosa, Valber A

    2014-11-01

    We describe the amperometric detection of glucose using oriented nanowires with magnetic switching of the bioelectrochemical process. The fabrication process of the nanowires was prepared through controlled nucleation and growth during a stepwise electrochemical deposition, and it was characterized using scanning electron microscopy. Cyclic voltammetry and amperometry were used to study the magnetoswitchable property; this control was accomplished by changing the surface orientation of nanowires. Under the optimal condition, the amperometric response was also linear up to a glucose concentration of 0.1-16.0 mmol L(-1) with a sensitivity of 81 μA mM(-1). The detection limit was estimated for 4.8×10(-8) mol L(-1), defined from a signal/noise ratio of 3. It also exhibits good reproducibility and high selectivity with insignificant interference from ascorbic acid, acetoaminophen, and uric acid. The resulting biosensor was applied to detect the blood sugar in human serum samples without any pretreatment, and the results were comparatively in agreement with the clinical assay.

  11. Glucocorticoid receptor exhibits sexually dimorphic expression in the medaka brain.

    PubMed

    Kikuchi, Yukiko; Hosono, Kohei; Yamashita, Junpei; Kawabata, Yukika; Okubo, Kataaki

    2015-11-01

    The differential impact of stress on brain functions of males and females has been widely observed in vertebrates. Recent evidence suggests that stress-induced glucocorticoid signaling affects sexual differentiation and sex changes in teleost fish. These facts led us to postulate that there were sex differences in glucocorticoid signaling in the teleost brain that underlie some sex differences in their physiological and behavioral traits. Here we found sexually dimorphic expression of a glucocorticoid receptor gene (gr1) in the brain of medaka fish (Oryzias latipes), with females having greater expression in several preoptic and thalamic nuclei. Further, gr1 exhibits female-biased expression in neurons of the anterior parvocellular preoptic nucleus that produce the neuropeptides vasotocin and gonadotropin-releasing hormone 1 (these neuropeptides have been implicated in the regulation of neuroendocrine and behavioral functions). These findings suggest that glucocorticoids have a greater influence on physiology and behavior mediated by these neuropeptides in females than in males, which may contribute to sex differences in the brain's response to stress. PMID:26433060

  12. Morphology development in polymer blends exhibiting strong intermolecular interactions

    SciTech Connect

    Weiss, R.A.; Feng, Y.; Han, C.C.; Karim, A.

    1996-12-31

    He et al. measured the spinodal decomposition (SD) kinetics of a blend of poly(butyl methacrylate) with a polystyrene modified with 1.5 mol% of a hydroxy-containing comonomer that exhibited intermolecular hydrogen bonding, who studied blends. For small excursions into the spinodal region, multiple structures developed in the blend, which suggested that multiple mechanisms may be involved in the phase separation process. For most cases, however, the kinetics of phase separation followed Cahn-Hilliard theory in the early stage of spinodal decomposition and a self-similar mechanism in the later stages, similar to non-associating polymer blends such as PS/PVME. The failure to observe an effect of a specific intermolecular interaction on SD kinetics may be a consequence of the low concentration of hydroxyl groups on the polystyrene, ca. five per chain, and the weakening of the hydrogen bond at the elevated temperatures used to study phase separation where the crosslink effect of hydrogen bonding may not be significant. At elevated temperatures, the association-dissociation equilibrium shifts towards non-associated hydroxyl and ester groups. An objective of the present study was to use a polymer blend having a relatively higher degree of intermolecular association at the phase separation temperature and to investigate how physical crosslinks affect the phase separation kinetics accompanying spinodal decomposition.

  13. Respiratory-deficient human fibroblasts exhibiting defective mitochondrial DNA replication.

    PubMed Central

    Bodnar, A G; Cooper, J M; Leonard, J V; Schapira, A H

    1995-01-01

    We have characterized cultured skin fibroblasts from two siblings affected with a fatal mitochondrial disease caused by a nuclear genetic defect. Mitochondrial respiratory-chain function was severely decreased in these cells. Southern-blot analysis showed that the fibroblasts had reduced levels of mitochondrial DNA (mtDNA). The mtDNA was unstable and was eliminated from the cultured cells over many generations, generating the rho0 genotype. As the mtDNA level decreased, the cells became more dependent upon pyruvate and uridine for growth. Nuclear-encoded subunits of respiratory-chain complexes were synthesized and imported into the mitochondria of the mtDNA-depleted cells, albeit at reduced levels compared with the controls. Mitochondrial protein synthesis directed by the residual mtDNA indicated that the mtDNA was expressed and that the defect specifically involves the replication or maintenance of mtDNA. This is a unique example of a respiratory-deficient human cell line exhibiting defective mtDNA replication. Images Figure 1 Figure 2 Figure 4 Figure 5 PMID:7848281

  14. Power gain exhibited by motile mechanosensory neurons in Drosophila ears

    PubMed Central

    Göpfert, M. C.; Humphris, A. D. L.; Albert, J. T.; Robert, D.; Hendrich, O.

    2005-01-01

    In insects and vertebrates alike, hearing is assisted by the motility of mechanosensory cells. Much like pushing a swing augments its swing, this cellular motility is thought to actively augment vibrations inside the ear, thus amplifying the ear's mechanical input. Power gain is the hallmark of such active amplification, yet whether and how much energy motile mechanosensory cells contribute within intact auditory systems has remained uncertain. Here, we assess the mechanical energy provided by motile mechanosensory neurons in the antennal hearing organs of Drosophila melanogaster by analyzing the fluctuations of the sound receiver to which these neurons connect. By using dead WT flies and live mutants (tilB2, btv5P1, and nompA2) with defective neurons as a background, we show that the intact, motile neurons do exhibit power gain. In WT flies, the neurons lift the receiver's mean total energy by 19 zJ, which corresponds to 4.6 times the energy of the receiver's Brownian motion. Larger energy contributions (200 zJ) associate with self-sustained oscillations, suggesting that the neurons adjust their energy expenditure to optimize the receiver's sensitivity to sound. We conclude that motile mechanosensory cells provide active amplification; in Drosophila, mechanical energy contributed by these cells boosts the vibrations that enter the ear. PMID:15623551

  15. Power gain exhibited by motile mechanosensory neurons in Drosophila ears.

    PubMed

    Göpfert, M C; Humphris, A D L; Albert, J T; Robert, D; Hendrich, O

    2005-01-11

    In insects and vertebrates alike, hearing is assisted by the motility of mechanosensory cells. Much like pushing a swing augments its swing, this cellular motility is thought to actively augment vibrations inside the ear, thus amplifying the ear's mechanical input. Power gain is the hallmark of such active amplification, yet whether and how much energy motile mechanosensory cells contribute within intact auditory systems has remained uncertain. Here, we assess the mechanical energy provided by motile mechanosensory neurons in the antennal hearing organs of Drosophila melanogaster by analyzing the fluctuations of the sound receiver to which these neurons connect. By using dead WT flies and live mutants (tilB(2), btv(5P1), and nompA(2)) with defective neurons as a background, we show that the intact, motile neurons do exhibit power gain. In WT flies, the neurons lift the receiver's mean total energy by 19 zJ, which corresponds to 4.6 times the energy of the receiver's Brownian motion. Larger energy contributions (200 zJ) associate with self-sustained oscillations, suggesting that the neurons adjust their energy expenditure to optimize the receiver's sensitivity to sound. We conclude that motile mechanosensory cells provide active amplification; in Drosophila, mechanical energy contributed by these cells boosts the vibrations that enter the ear. PMID:15623551

  16. The leatherback turtle, Dermochelys coriacea, exhibits both polyandry and polygyny.

    PubMed

    Crim, J L; Spotila, L D; Spotila, J R; O'Connor, M; Reina, R; Williams, C J; Paladino, F V

    2002-10-01

    The leatherback turtle (Dermochelys coriacea) is an endangered species, and world-wide populations are declining. To understand better the mating structure of this pelagic and fragile species, we investigated paternity in nearly 1000 hatchlings from Playa Grande in Parque Marino Nacional Las Baulas, Costa Rica. We collected DNA samples from 36 adult female leatherbacks and assessed allele frequency distributions for three microsatellite loci. For 20 of these 36 females, we examined DNA from hatchlings representing multiple clutches, and in some cases assessed up to four successive clutches from the same female. We inferred paternal alleles by comparing maternal and hatchling genotypes. We could not reject the null hypothesis of single paternity in 12 of 20 families (31 of 50 clutches), but we did reject the null hypothesis in two families (eight of 50 clutches). In the remaining six families, the null hypothesis could not be accepted or rejected with certainty because the number of hatchlings exhibiting extra nonmaternal alleles was small, and could thus be a result of mutation or sample error. Successive clutches laid by the same female had the same paternal allelic contribution, indicating sperm storage or possibly monogamy. None of 20 females shared the same three-locus genotype whereas there were two instances of shared genotypes among 17 inferred paternal three-locus genotypes. We conclude that both polyandry and polygyny are part of the mating structure of this leatherback sea turtle population.

  17. Magnetic tweezers force calibration for molecules that exhibit conformational switching

    NASA Astrophysics Data System (ADS)

    Jacobson, David R.; Saleh, Omar A.

    2016-09-01

    High spatial and temporal resolution magnetic tweezers experiments allow for the direct calibration of pulling forces applied to short biomolecules. In one class of experiments, a force is applied to a structured RNA or protein to induce an unfolding transition; when the force is maintained at particular values, the molecule can exhibit conformational switching between the folded and unfolded states or between intermediate states. Here, we analyze the degree to which common force calibration approaches, involving the fitting of model functions to the Allan variance or power spectral density of the bead trajectory, are biased by this conformational switching. We find significant effects in two limits: that of large molecular extension changes between the two states, in which alternative fitting functions must be used, and that of very fast switching kinetics, in which the force calibration cannot be recovered due to the slow diffusion time of the magnetic bead. We use simulations and high-resolution RNA hairpin data to show that most biophysical experiments do not occur in either of these limits.

  18. Artificially engineered Heusler ferrimagnetic superlattice exhibiting perpendicular magnetic anisotropy.

    PubMed

    Ma, Q L; Zhang, X M; Miyazaki, T; Mizukami, S

    2015-01-01

    To extend density limits in magnetic recording industry, two separate strategies were developed to build the storage bit in last decade, introduction of perpendicular magnetic anisotropy (PMA) and adoption of ferrimagnetism/antiferromagnetism. Meanwhile, these properties significantly improve device performance, such as reducing spin-transfer torque energy consumption and decreasing signal-amplitude-loss. However, materials combining PMA and antiferromagnetism rather than transition-metal/rare-earth system were rarely developed. Here, we develop a new type of ferrimagnetic superlattice exhibiting PMA based on abundant Heusler alloy families. The superlattice is formed by [MnGa/Co2FeAl] unit with their magnetizations antiparallel aligned. The effective anisotropy (K(u)(eff)) over 6 Merg/cm(3) is obtained, and the SL can be easily built on various substrates with flexible lattice constants. The coercive force, saturation magnetization and K(u)(eff) of SLs are highly controllable by varying the thickness of MnGa and Co2FeAl layers. The SLs will supply a new choice for magnetic recording and spintronics memory application such as magnetic random access memory. PMID:25597496

  19. Mechanical Amplification Exhibited by Quiescent Saccular Hair Bundles

    PubMed Central

    Roongthumskul, Yuttana; Bozovic, Dolores

    2015-01-01

    Spontaneous oscillations exhibited by free-standing hair bundles from the Bullfrog sacculus suggest the existence of an active process that might underlie the exquisite sensitivity of the sacculus to mechanical stimulation. However, this spontaneous activity is suppressed by coupling to an overlying membrane, which applies a large mechanical load on the bundle. How a quiescent hair bundle utilizes its active process is still unknown. We studied the dynamics of motion of individual hair bundles under different offsets in the bundle position, and observed the occurrence of spikes in hair-bundle motion, associated with the generation of active work. These mechanical spikes can be evoked by a sinusoidal stimulus, leading to an amplified movement of the bundle with respect to the passive response. Amplitude gain reached as high as 100-fold at small stimulus amplitudes. Amplification of motion decreased with increasing amplitude of stimulation, ceasing at ∼6–12 pN stimuli. Results from numerical simulations suggest that the adaptation process, mediated by myosin 1c, is not required for the production of mechanical spikes. PMID:25564852

  20. Mechanical amplification exhibited by quiescent saccular hair bundles.

    PubMed

    Roongthumskul, Yuttana; Bozovic, Dolores

    2015-01-01

    Spontaneous oscillations exhibited by free-standing hair bundles from the Bullfrog sacculus suggest the existence of an active process that might underlie the exquisite sensitivity of the sacculus to mechanical stimulation. However, this spontaneous activity is suppressed by coupling to an overlying membrane, which applies a large mechanical load on the bundle. How a quiescent hair bundle utilizes its active process is still unknown. We studied the dynamics of motion of individual hair bundles under different offsets in the bundle position, and observed the occurrence of spikes in hair-bundle motion, associated with the generation of active work. These mechanical spikes can be evoked by a sinusoidal stimulus, leading to an amplified movement of the bundle with respect to the passive response. Amplitude gain reached as high as 100-fold at small stimulus amplitudes. Amplification of motion decreased with increasing amplitude of stimulation, ceasing at ∼6-12 pN stimuli. Results from numerical simulations suggest that the adaptation process, mediated by myosin 1c, is not required for the production of mechanical spikes. PMID:25564852