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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

    DOEpatents

    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. Purification and characterization of an alpha-glucosidase from a hyperthermophilic archaebacterium, Pyrococcus furiosus, exhibiting a temperature optimum of 105 to 115 degrees C.

    PubMed Central

    Costantino, H R; Brown, S H; Kelly, R M

    1990-01-01

    Pyrococcus furiosus is a strictly anaerobic hyperthermophilic archaebacterium with an optimal growth temperature of about 100 degrees C. When this organism was grown in the presence of certain complex carbohydrates, the production of several amylolytic enzymes was noted. These enzymes included an alpha-glucosidase that was located in the cell cytoplasm. This alpha-glucosidase has been purified 310-fold and corresponded to a protein band of 125 kilodaltons as resolved by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme exhibited optimum activity at pH 5.0 to 6.0 and over a temperature range of 105 to 115 degrees C. Kinetic analysis conducted at 108 degrees C revealed hydrolysis of the substrates p-nitrophenyl-alpha-D-glucopyranoside (PNPG), methyl-alpha-D-glucopyranoside, maltose, and isomaltose. Trace activity was detected towards p-nitrophenyl-beta-D-glucopyranoside, and no activity could be detected towards starch or sucrose. Inhibition studies conducted at 108 degrees C with PNPG as the substrate and maltose as the inhibitor yielded a Ki for maltose of 14.3 mM. Preincubation for 30 min at 98 degrees C in 100 mM dithiothreitol and 1.0 M urea had little effect on enzyme activity, whereas preincubation in 1.0% sodium dodecyl sulfate and 1.0 M guanidine hydrochloride resulted in significant loss of enzyme activity. Purified alpha-glucosidase from P. furiosus exhibited remarkable thermostability; incubation of the enzyme at 98 degrees C resulted in a half life of nearly 48 h. Images PMID:2163383

  4. 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.

  5. Structure of hyperthermophilic β-glucosidase from Pyrococcus furiosus.

    PubMed

    Kado, Yuji; Inoue, Tsuyoshi; Ishikawa, Kazuhiko

    2011-12-01

    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

  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. Expression, Purification, and Characterisation of Dehydroquinate Synthase from Pyrococcus furiosus

    PubMed Central

    Negron, Leonardo; Patchett, Mark L.; Parker, Emily J.

    2011-01-01

    Dehydroquinate synthase (DHQS) catalyses the second step of the shikimate pathway to aromatic compounds. DHQS from the archaeal hyperthermophile Pyrococcus furiosus was insoluble when expressed in Escherichia coli but was partially solubilised when KCl was included in the cell lysis buffer. A purification procedure was developed, involving lysis by sonication at 30°C followed by a heat treatment at 70°C and anion exchange chromatography. Purified recombinant P. furiosus DHQS is a dimer with a subunit Mr of 37,397 (determined by electrospray ionisation mass spectrometry) and is active over broad pH and temperature ranges. The kinetic parameters are KM (3-deoxy-D-arabino-heptulosonate 7-phosphate) 3.7 μM and kcat 3.0 sec−1 at 60°C and pH 6.8. EDTA inactivates the enzyme, and enzyme activity is restored by several divalent metal ions including (in order of decreasing effectiveness) Cd2+, Co2+, Zn2+, and Mn2+. High activity of a DHQS in the presence of Cd2+ has not been reported for enzymes from other sources, and may be related to the bioavailability of Cd2+ for P. furiosus. This study is the first biochemical characterisation of a DHQS from a thermophilic source. Furthermore, the characterisation of this hyperthermophilic enzyme was carried out at elevated temperatures using an enzyme-coupled assay. PMID:21603259

  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

    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

  10. Changes in the catalytic properties of Pyrococcus furiosus thermostable amylase by mutagenesis of the substrate binding sites.

    PubMed

    Yang, Sung-Jae; Min, Byoung-Chul; Kim, Young-Wan; Jang, Sang-Mok; Lee, Byong-Hoon; Park, Kwan-Hwa

    2007-09-01

    Pyrococcus furiosus thermostable amylase (TA) is a cyclodextrin (CD)-degrading enzyme with a high preference for CDs over maltooligosaccharides. In this study, we investigated the roles of four residues (His414, Gly415, Met439, and Asp440) in the function of P. furiosus TA by using site-directed mutagenesis and kinetic analysis. A variant form of P. furiosus TA containing two mutations (H414N and G415E) exhibited strongly enhanced alpha-(1,4)-transglycosylation activity, resulting in the production of a series of maltooligosaccharides that were longer than the initial substrates. In contrast, the variant enzymes with single mutations (H414N or G415E) showed a substrate preference similar to that of the wild-type enzyme. Other mutations (M439W and D440H) reversed the substrate preference of P. furiosus TA from CDs to maltooligosaccharides. Relative substrate preferences for maltoheptaose over beta-CD, calculated by comparing k(cat)/K(m) ratios, of 1, 8, and 26 for wild-type P. furiosus TA, P. furiosus TA with D440H, and P. furiosus TA with M439W and D440H, respectively, were found. Our results suggest that His414, Gly415, Met439, and Asp440 play important roles in substrate recognition and transglycosylation. Therefore, this study provides information useful in engineering glycoside hydrolase family 13 enzymes. PMID:17630303

  11. Neutron diffraction studies on rubredoxin from Pyrococcus furiosus.

    PubMed

    Bau, Robert

    2004-01-01

    Single-crystal neutron diffraction data up to a resolution of 1.5 A have been collected at room temperature on two forms of rubredoxin using the BIX-3 diffractometer at the JRR-3 reactor of the Japan Atomic Energy Research Institute (JAERI). Rubredoxin is a small iron-sulfur redox protein with 53 amino acid residues, and the source of this particular protein is the hyperthermophile Pyrococcus furiosus, a microorganism that normally lives at temperatures near that of boiling water. Data were collected on crystals of the wild-type protein and on a mutant in which three of the residues have been replaced. In this paper we will be describing several sets of results arising from these high-resolution neutron structure determinations: (a) the H/D exchange pattern of the N-H bonds of the main backbone, which give information about which regions of the molecule are more exposed to solvent; (b) the orientations of some of the O-D bonds in the protein, information which is often not obtainable from X-ray results; (c) the structure and appearance of water molecules in the protein crystals; and (d) some structural features which may help rationalize the remarkable thermal stability of the wild-type protein from this intriguing microorganism PMID:14646139

  12. 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

  13. 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.

  14. 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

  15. Complete saccharification of β-glucan using hyperthermophilic endocellulase and β-glucosidase from Pyrococcus furiosus.

    PubMed

    Kataoka, Misumi; Ishikawa, Kazuhiko

    2014-01-01

    Hyperthermophilic cellulase is an industrially important enzyme for biomass saccharification at high temperature. Two hyperthermophilic cellulases from the hyperthermophile Pyrococcus furiosus, endocellulase (EGPf) and β-glucosidase (BGLPf), exhibit optimal activity at 90-105 °C and a combination of two enzymes can hydrolyze a wide range of β-linked substrates. EGPf cleaves the β(1→4) bond of various substrates containing either only the β(1→4) linkage or β(1→3),(1→4) mixed-linkages. In contrast, BGLPf preferentially hydrolyzes the β(1→3) linkage over the β(1→4) linkage of disaccharides. β-Glucans are polysaccharides of D-glucose monomers formed by β(1→3),(1→4) mixed-linkage bonds. They occur most commonly as cellulose in plants, in the bran of cereal grains, the cell wall of baker's yeast, and in certain fungi, mushrooms, and bacteria. We reveal that β-glucan can be completely degraded to glucose at high temperature with a combination of EGPf and BGLPf. PMID:25209501

  16. A novel endonuclease that may be responsible for damaged DNA base repair in Pyrococcus furiosus

    PubMed Central

    Shiraishi, Miyako; Ishino, Sonoko; Yamagami, Takeshi; Egashira, Yuriko; Kiyonari, Shinichi; Ishino, Yoshizumi

    2015-01-01

    DNA is constantly damaged by endogenous and environmental influences. Deaminated adenine (hypoxanthine) tends to pair with cytosine and leads to the A:T→G:C transition mutation during DNA replication. Endonuclease V (EndoV) hydrolyzes the second phosphodiester bond 3′ from deoxyinosine in the DNA strand, and was considered to be responsible for hypoxanthine excision repair. However, the downstream pathway after EndoV cleavage remained unclear. The activity to cleave the phosphodiester bond 5′ from deoxyinosine was detected in a Pyrococcus furiosus cell extract. The protein encoded by PF1551, obtained from the mass spectrometry analysis of the purified fraction, exhibited the corresponding cleavage activity. A putative homolog from Thermococcus kodakarensis (TK0887) showed the same activity. Further biochemical analyses revealed that the purified PF1551 and TK0887 proteins recognize uracil, xanthine and the AP site, in addition to hypoxanthine. We named this endonuclease Endonuclease Q (EndoQ), as it may be involved in damaged base repair in the Thermococcals of Archaea. PMID:25694513

  17. 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.

  18. 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

  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

    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

  1. 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

  2. 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. PMID:25817554

  3. Structure of a double hexamer of the Pyrococcus furiosus minichromosome maintenance protein N-terminal domain.

    PubMed

    Meagher, Martin; Enemark, Eric J

    2016-07-01

    The crystal structure of the N-terminal domain of the Pyrococcus furiosus minichromosome maintenance (MCM) protein as a double hexamer is described. The MCM complex is a ring-shaped helicase that unwinds DNA at the replication fork of eukaryotes and archaea. Prior to replication initiation, the MCM complex assembles as an inactive double hexamer at specific sites of DNA. The presented structure is highly consistent with previous MCM double-hexamer structures and shows two MCM hexamers with a head-to-head interaction mediated by the N-terminal domain. Minor differences include a diminished head-to-head interaction and a slightly reduced inter-hexamer rotation. PMID:27380371

  4. Functional reconstitution and characterization of Pyrococcus furiosus RNase P

    PubMed Central

    Tsai, Hsin-Yue; Pulukkunat, Dileep K.; Woznick, Walter K.; Gopalan, Venkat

    2006-01-01

    RNase P, which catalyzes the magnesium-dependent 5′-end maturation of tRNAs in all three domains of life, is composed of one essential RNA and a varying number of protein subunits depending on the source: at least one in bacteria, four in archaea, and nine in eukarya. To address why multiple protein subunits are needed for archaeal/eukaryal RNase P catalysis, in contrast to their bacterial relative, in vitro reconstitution of these holoenzymes is a prerequisite. Using recombinant subunits, we have reconstituted in vitro the RNase P holoenzyme from the thermophilic archaeon Pyroccocus furiosus (Pfu) and furthered our understanding regarding its functional organization and assembly pathway(s). Whereas Pfu RNase P RNA (RPR) alone is capable of multiple turnover, addition of all four RNase P protein (Rpp) subunits to Pfu RPR results in a 25-fold increase in its kcat and a 170-fold decrease in Km. In fact, even in the presence of only one of two specific pairs of Rpps, the RPR displays activity at lower substrate and magnesium concentrations. Moreover, a pared-down, mini-Pfu RNase P was identified with an RPR deletion mutant. Results from our kinetic and footprinting studies on Pfu RNase P, together with insights from recent structures of bacterial RPRs, provide a framework for appreciating the role of multiple Rpps in archaeal RNase P. PMID:17053064

  5. 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

  6. Overexpression, purification and crystallization of an archaeal DNA ligase from Pyrococcus furiosus

    SciTech Connect

    Nishida, Hirokazu; Tsuchiya, Daisuke; Ishino, Yoshizumi; Morikawa, Kosuke

    2005-12-01

    Crystals of the archaeal DNA ligase from Pyrococcus furiosus were obtained using 6.6%(v/v) ethanol as a precipitant and diffracted X-rays to 1.7 Å resolution. DNA ligases seal single-strand breaks in double-stranded DNA and their function is essential to maintain the integrity of the genome during various aspects of DNA metabolism, such as replication, excision repair and recombination. DNA-strand breaks are frequently generated as reaction intermediates in these events and the sealing of these breaks depends solely on the proper function of DNA ligase. Crystals of the archaeal DNA ligase from Pyrococcus furiosus were obtained using 6.6%(v/v) ethanol as a precipitant and diffracted X-rays to 1.7 Å resolution. They belong to the monoclinic space group P2{sub 1}, with unit-cell parameters a = 61.1, b = 88.3, c = 63.4 Å, β = 108.9°. The asymmetric unit contains one ligase molecule.

  7. 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

  8. 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

  9. 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.

  10. Bioenergetic studies of sulfur reduction in the hyperthermophilic archaebacteria Pyrodictium brockii and Pyrococcus furiosus

    SciTech Connect

    Schicho, R.N.

    1992-01-01

    The central focus of this work was the investigation of the central energy generating pathways of two hyperthermophilic sulfidogenic archaebacteria, Pyrodictium brockii and Pyrococcus furiosus. A potential application of these organisms in the desulfurization of coals was investigated. An effective ;means of removing elemental sulfur (S[degrees]) was developed. Analytical and processing applications are discussed. The rates of sulfur removal by the hyperthermophiles were 5 fold those measured of the mesophile, Thiobacillus thiooxidans. The primary energy generating pathway of Pyrodicutium brockii has been termed hydrogen-sulfur autotrophy and is characterized by the oxidation of H[sub 2] and reduction of S[degrees]. The goals of this part of the work were to quantify the stoichiometry of this organism and to estimate Y[sub s[sup MAX

  11. 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

  12. Molecular and biochemical characterization of the ADP-dependent phosphofructokinase from the hyperthermophilic archaeon Pyrococcus furiosus.

    PubMed

    Tuininga, J E; Verhees, C H; van der Oost, J; Kengen, S W; Stams, A J; de Vos, W M

    1999-07-23

    Pyrococcus furiosus uses a modified Embden-Meyerhof pathway involving two ADP-dependent kinases. Using the N-terminal amino acid sequence of the previously purified ADP-dependent glucokinase, the corresponding gene as well as a related open reading frame were detected in the genome of P. furiosus. Both genes were successfully cloned and expressed in Escherichia coli, yielding highly thermoactive ADP-dependent glucokinase and phosphofructokinase. The deduced amino acid sequences of both kinases were 21.1% identical but did not reveal significant homology with those of other known sugar kinases. The ADP-dependent phosphofructokinase was purified and characterized. The oxygen-stable protein had a native molecular mass of approximately 180 kDa and was composed of four identical 52-kDa subunits. It had a specific activity of 88 units/mg at 50 degrees C and a pH optimum of 6.5. As phosphoryl group donor, ADP could be replaced by GDP, ATP, and GTP to a limited extent. The K(m) values for fructose 6-phosphate and ADP were 2.3 and 0.11 mM, respectively. The phosphofructokinase did not catalyze the reverse reaction, nor was it regulated by any of the known allosteric modulators of ATP-dependent phosphofructokinases. ATP and AMP were identified as competitive inhibitors of the phosphofructokinase, raising the K(m) for ADP to 0.34 and 0.41 mM, respectively. PMID:10409652

  13. Anaerobic desulfurization of ground rubber with the thermophilic archaeon Pyrococcus furiosus--a new method for rubber recycling.

    PubMed

    Bredberg, K; Persson, J; Christiansson, M; Stenberg, B; Holst, O

    2001-01-01

    The anaerobic sulfur-reducing archaeon Pyrococcus furiosus was investigated regarding its capacity to desulfurize rubber material. The microorganism's sensitivity towards common rubber elastomers and additives was tested and several were shown to be toxic to P. furiosus. The microorganism was shown to utilize sulfur in vulcanized natural rubber and an increase in cell density was obtained when cultivated in the presence of spent tire rubber. Ethanol-leached cryo-ground tire rubber treated with P. furiosus for 10 days was vulcanized together with virgin rubber material (15% w/w) and the mechanical properties of the resulting material were determined. The increase in the stress at break value and the decrease in swell ratio and stress relaxation rate obtained for material containing microbially treated rubber (compared to untreated material) show the positive effects of microbial desulfurization on rubber. PMID:11234957

  14. 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

  15. 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.

  16. Purification and characterization of a cobalt-activated carboxypeptidase from the hyperthermophilic archaeon Pyrococcus furiosus.

    PubMed Central

    Cheng, T. C.; Ramakrishnan, V.; Chan, S. I.

    1999-01-01

    A novel metallocarboxypeptidase (PfuCP) has been purified to homogeneity from the hyperthermophilic archaeon, Pyrococcus furiosus, with its intended use in C-terminal ladder sequencing of proteins and peptides at elevated temperatures. PfuCP was purified in its inactive state by the addition of ethylenediaminetetraacetic acid (EDTA) and dithiothreitol (DTT) to purification buffers, and the activity was restored by the addition of divalent cobalt (K, = 24 +/- 4 microM at 80 degrees C). The serine protease inhibitor phenylmethylsulfonyl fluoride (PMSF) had no effect on the activity. The molecular mass of monomeric PfuCP is 59 kDa as determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and 58 kDa by SDS-PAGE analysis. In solution, PfuCP exists as a homodimer of approximately 128 kDa as determined by gel filtration chromatography. The activity of PfuCP exhibits a temperature optimum exceeding 90 degrees C under ambient pressure, and a narrow pH optimum of 6.2-6.6. Addition of Co2+ to the apoPfuCP at room temperature does not alter its far-UV circular dichroism (CD) or its intrinsic fluorescence spectrum. Even when the CoPfuCP is heated to 80 degrees C, its far-UV CD shows a minimal change in the global conformation and the intrinsic fluorescence of aromatic residues shows only a partial quenching. Changes in the intrinsic fluorescence appear essentially reversible with temperature. Finally, the far-UV CD and intrinsic fluorescence data suggest that the overall structure of the holoenzyme is extremely thermostable. However, the activities of both the apo and holo enzyme exhibit a similar second-order decay over time, with 50% activity remaining after approximately 40 min at 80 degrees C. The N-blocked synthetic dipeptide, N-carbobenzoxy-Ala-Arg (ZAR), was used in the purification assay. The kinetic parameters at 80 degrees C with 0.4 mM CoCl2 were: Km, 0.9 +/- 0.1 mM; Vmax, 2,300 +/- 70 U mg(-1); and turn over number

  17. 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.

  18. Multiple crystal forms of N,N'-diacetylchitobiose deacetylase from Pyrococcus furiosus.

    PubMed

    Nakamura, Tsutomu; Niiyama, Mayumi; Hashimoto, Wakana; Ida, Kurumi; Abe, Manabu; Morita, Junji; Uegaki, Koichi

    2015-06-01

    Native N,N'-diacetylchitobiose deacetylase from Pyrococcus furiosus (Pf-Dac) and its selenomethionine derivative (Se-Pf-Dac) were crystallized and analyzed in the presence and absence of cadmium ion. The four crystal structures fell into three different crystal-packing groups, with the cadmium-free Pf-Dac and Se-Pf-Dac belonging to the same space group, with homologous unit-cell parameters. The crystal structures in the presence of cadmium contained distorted octahedral cadmium complexes coordinated by three chlorides, two O atoms and an S or Se atom from the N-terminal methionine or selenomethionine, respectively. The N-terminal cadmium complex was involved in crystal contacts between symmetry-related molecules through hydrogen bonding to the N-termini. While all six N-termini of Se-Pf-Dac were involved in cadmium-complex formation, only two of the Pf-Dac N-termini participated in complex formation in the Cd-containing crystal, resulting in different crystal forms. These differences are discussed in light of the higher stability of the Cd-Se bond than the Cd-S bond. This work provides an example of the contribution of cadmium towards determining protein crystal quality and packing depending on the use of the native protein or the selenomethionine derivative. PMID:26057790

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

    PubMed

    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 Ca(2+) for activity or thermostability, and >80% of the enzyme activity was retained after treatment at 100 °C for 4 h. PMID:26916714

  20. 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

  1. 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.

  2. 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

  3. Flagella of Pyrococcus furiosus: multifunctional organelles, made for swimming, adhesion to various surfaces, and cell-cell contacts.

    PubMed

    Näther, Daniela J; Rachel, Reinhard; Wanner, Gerhard; Wirth, Reinhard

    2006-10-01

    Pyrococcus furiosus ("rushing fireball") was named for the ability of this archaeal coccus to rapidly swim at its optimal growth temperature, around 100 degrees C. Early electron microscopic studies identified up to 50 cell surface appendages originating from one pole of the coccus, which have been called flagella. We have analyzed these putative motility organelles and found them to be composed primarily (>95%) of a glycoprotein that is homologous to flagellins from other archaea. Using various electron microscopic techniques, we found that these flagella can aggregate into cable-like structures, forming cell-cell connections between ca. 5% of all cells during stationary growth phase. P. furiosus cells could adhere via their flagella to carbon-coated gold grids used for electron microscopic analyses, to sand grains collected from the original habitat (Porto di Levante, Vulcano, Italy), and to various other surfaces. P. furiosus grew on surfaces in biofilm-like structures, forming microcolonies with cells interconnected by flagella and adhering to the solid supports. Therefore, we concluded that P. furiosus probably uses flagella for swimming but that the cell surface appendages also enable this archaeon to form cable-like cell-cell connections and to adhere to solid surfaces. PMID:16980494

  4. Relationship between Glycosyl Hydrolase Inventory and Growth Physiology of the Hyperthermophile Pyrococcus furiosus on Carbohydrate-Based Media

    PubMed Central

    Driskill, Lance E.; Kusy, Kevin; Bauer, Michael W.; Kelly, Robert M.

    1999-01-01

    Utilization of a range of carbohydrates for growth by the hyperthermophile Pyrococcus furiosus was investigated by examining the spectrum of glycosyl hydrolases produced by this microorganism and the thermal labilities of various saccharides. Previously, P. furiosus had been found to grow in batch cultures on several α-linked carbohydrates and cellobiose but not on glucose or other β-linked sugars. Although P. furiosus was not able to grow on any nonglucan carbohydrate or any form of cellulose in this study (growth on oat spelt arabinoxylan was attributed to glucan contamination of this substrate), significant growth at 98°C occurred on β-1,3- and β-1,3–β-1,4-linked glucans. Oligosaccharides generated by digestion with a recombinant laminarinase derived from P. furiosus were the compounds that were most effective in stimulating growth of the microorganism. In several cases, periodic addition of β-glucan substrates to fed-batch cultures limited adverse thermochemical modifications of the carbohydrates (i.e., Maillard reactions and caramelization) and led to significant increases (as much as two- to threefold) in the cell yields. While glucose had only a marginally positive effect on growth in batch culture, the final cell densities nearly tripled when glucose was added by the fed-batch procedure. Nonenzymatic browning reactions were found to be significant at 98°C for saccharides with degrees of polymerization (DP) ranging from 1 to 6; glucose was the most labile compound on a mass basis and the least labile compound on a molar basis. This suggests that for DP of 2 or greater protection of the nonreducing monosaccharide component may be a factor in substrate availability. For P. furiosus, carbohydrate utilization patterns were found to reflect the distribution of the glycosyl hydrolases which are known to be produced by this microorganism. PMID:10049838

  5. Relationship between glycosyl hydrolase inventory and growth physiology of the hyperthermophile Pyrococcus furiosus on carbohydrate-based media.

    PubMed

    Driskill, L E; Kusy, K; Bauer, M W; Kelly, R M

    1999-03-01

    Utilization of a range of carbohydrates for growth by the hyperthermophile Pyrococcus furiosus was investigated by examining the spectrum of glycosyl hydrolases produced by this microorganism and the thermal labilities of various saccharides. Previously, P. furiosus had been found to grow in batch cultures on several alpha-linked carbohydrates and cellobiose but not on glucose or other beta-linked sugars. Although P. furiosus was not able to grow on any nonglucan carbohydrate or any form of cellulose in this study (growth on oat spelt arabinoxylan was attributed to glucan contamination of this substrate), significant growth at 98 degrees C occurred on beta-1,3- and beta-1,3-beta-1,4-linked glucans. Oligosaccharides generated by digestion with a recombinant laminarinase derived from P. furiosus were the compounds that were most effective in stimulating growth of the microorganism. In several cases, periodic addition of beta-glucan substrates to fed-batch cultures limited adverse thermochemical modifications of the carbohydrates (i.e., Maillard reactions and caramelization) and led to significant increases (as much as two- to threefold) in the cell yields. While glucose had only a marginally positive effect on growth in batch culture, the final cell densities nearly tripled when glucose was added by the fed-batch procedure. Nonenzymatic browning reactions were found to be significant at 98 degrees C for saccharides with degrees of polymerization (DP) ranging from 1 to 6; glucose was the most labile compound on a mass basis and the least labile compound on a molar basis. This suggests that for DP of 2 or greater protection of the nonreducing monosaccharide component may be a factor in substrate availability. For P. furiosus, carbohydrate utilization patterns were found to reflect the distribution of the glycosyl hydrolases which are known to be produced by this microorganism. PMID:10049838

  6. N-Linked Glycans Are Assembled on Highly Reduced Dolichol Phosphate Carriers in the Hyperthermophilic Archaea Pyrococcus furiosus

    PubMed Central

    Chang, Michelle M.; Imperiali, Barbara; Eichler, Jerry; Guan, Ziqiang

    2015-01-01

    In all three domains of life, N-glycosylation begins with the assembly of glycans on phosphorylated polyisoprenoid carriers. Like eukaryotes, archaea also utilize phosphorylated dolichol for this role, yet whereas the assembled oligosaccharide is transferred to target proteins from dolichol pyrophosphate in eukaryotes, archaeal N-linked glycans characterized to date are derived from a dolichol monophosphate carrier, apart from a single example. In this study, glycan-charged dolichol phosphate from the hyperthermophile Pyrococcus furiosus was identified and structurally characterized. Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated. PMID:26098850

  7. 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

  8. Key Role for Sulfur in Peptide Metabolism and in Regulation of Three Hydrogenases in the Hyperthermophilic Archaeon Pyrococcus furiosus

    PubMed Central

    Adams, Michael W. W.; Holden, James F.; Menon, Angeli Lal; Schut, Gerrit J.; Grunden, Amy M.; Hou, Chun; Hutchins, Andrea M.; Jenney, Francis E.; Kim, Chulhwan; Ma, Kesen; Pan, Guangliang; Roy, Roopali; Sapra, Rajat; Story, Sherry V.; Verhagen, Marc F. J. M.

    2001-01-01

    The hyperthermophilic archaeon Pyrococcus furiosus grows optimally at 100°C by the fermentation of peptides and carbohydrates. Growth of the organism was examined in media containing either maltose, peptides (hydrolyzed casein), or both as the carbon source(s), each with and without elemental sulfur (S0). Growth rates were highest on media containing peptides and S0, with or without maltose. Growth did not occur on the peptide medium without S0. S0 had no effect on growth rates in the maltose medium in the absence of peptides. Phenylacetate production rates (from phenylalanine fermentation) from cells grown in the peptide medium containing S0 with or without maltose were the same, suggesting that S0 is required for peptide utilization. The activities of 14 of 21 enzymes involved in or related to the fermentation pathways of P. furiosus were shown to be regulated under the five different growth conditions studied. The presence of S0 in the growth media resulted in decreases in specific activities of two cytoplasmic hydrogenases (I and II) and of a membrane-bound hydrogenase, each by an order of magnitude. The primary S0-reducing enzyme in this organism and the mechanism of the S0 dependence of peptide metabolism are not known. This study provides the first evidence for a highly regulated fermentation-based metabolism in P. furiosus and a significant regulatory role for elemental sulfur or its metabolites. PMID:11133967

  9. Purification, crystallization and preliminary X-ray diffraction studies of the soluble domain of the oligosaccharyltransferase STT3 subunit from the thermophilic archaeon Pyrococcus furiosus

    SciTech Connect

    Igura, Mayumi; Maita, Nobuo; Obita, Takayuki; Kamishikiryo, Jun; Maenaka, Katsumi; Kohda, Daisuke

    2007-09-01

    The C-terminal soluble domain of the catalytic subunit (STT3) of the oligosaccharyltransferase from P. furiosus was purified and crystallized. A native crystal and a SeMet derivative have been analyzed using X-ray diffraction. Oligosaccharyltransferase catalyzes the transfer of preassembled oligosaccharides onto asparagine residues in nascent polypeptide chains. The STT3 subunit is thought to bear the catalytic site. The C-terminal domain of the STT3 protein of Pyrococcus furiosus was expressed in Escherichia coli cells. STT3 protein prepared from two different sources, the soluble fraction and the inclusion bodies, produced crystals that diffracted to 2.7 Å. During crystallization screening, cocrystals of P. furiosus STT3 with an E. coli 50S ribosomal protein, L7/L12, were accidentally obtained. This cross-species interaction is not biologically relevant, but may be used to design a built-in polypeptide substrate for the STT3 crystals.

  10. Purification and characterization of two functional forms of intracellular protease PfpI from the hyperthermophilic archaeon Pyrococcus furiosus

    SciTech Connect

    Halio, S.B.; Bauer, M.W.; Kelley, R.M.

    1997-01-01

    The hyperthermophilic archaeon Pyrococcus furiosus grows optimally at 100{degrees}C by the fermentation of peptides and carbohydrates. From this organism, An intracellular protease was purified, previously designated PfpI (P. furiosus protease I). The protease contains exists in at least two functional conformations, which were purified separately. The predominant form from the purification (designated PfpI-C1) is a hexamer with a molecular mass of 124 {+-} 6 kDa (by gel filtration) and comprises about 90% of the total activity. The minor form (designated PfpI-C2) is trimeric with a molecular mass of 59 {+-} 3 kDa. PfpI-C1 hydrolyzed both basic and hydrophobic residues in the P1 position, indicating trypsin- and chymotrypsin-like specificities, respectively. The temperature optimum for Ala-Ala-Phe-7-amido-4-methylcoumarin (AAF-MCA) hydrolysis was {approximately}85{degrees}C both for purified PfpI-C1 and for proteolytic activity in P. furiosus cell extract. In contrast, the temperature optimum for PfpI prepared by incubating a cell extract of P. furiosus at 98{degrees}C in 1% sodium dodecyl sulfate for 24 h at 95 to 100{degrees}C, designated PfpI-H, was {approximately}100{degrees}C. Moreover, the half-life of activity of PfpI-C1 at 98{degrees}C was less than 30 min, in contrast to a value of more than 33 h measured for PfpI-H. PfpI-C1 appears to be a predominant serine-type protease in cell extracts but is converted in vitro, probably in part by deamination of Asn and Gln residues, to a more thermally stable form (PfpI-H) by prolonged heat treatment. The deamination hypothesis is supported by the differences in the measured pI values of PfpI-C1 (6.1) and PfpI-H (3.8). High levels of potassium phosphate (>0.5 mM) were found to extend the half-life of PfpI-C1 activity towards AAF-MCA by up to 2.5-fold at 90{degrees}C, suggesting that compatible solutes play an important role in the in vivo function of this protease. 43 refs., 6 figs., 2 tabs.

  11. 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

  12. Characterization of the Recombinant Thermostable Lipase (Pf2001) from Pyrococcus furiosus: Effects of Thioredoxin Fusion Tag and Triton X-100

    PubMed Central

    Alquéres, Sylvia Maria Campbell; Branco, Roberta Vieira; Freire, Denise Maria Guimarães; Alves, Tito Lívio Moitinho; Martins, Orlando Bonifácio; Almeida, Rodrigo Volcan

    2011-01-01

    In this work, the lipase from Pyrococcus furiosus encoded by ORF PF2001 was expressed with a fusion protein (thioredoxin) in Escherichia coli. The purified enzymes with the thioredoxin tag (TRX−PF2001Δ60) and without the thioredoxin tag (PF2001Δ60) were characterized, and various influences of Triton X-100 were determined. The optimal temperature for both enzymes was 80°C. Although the thioredoxin presence did not influence the optimum temperature, the TRX−PF2001Δ60 presented specific activity twice lower than the enzyme PF2001Δ60. The enzyme PF2001Δ60 was assayed using MUF-acetate, MUF-heptanoate, and MUF-palmitate. MUF-heptanoate was the preferred substrate of this enzyme. The chelators EDTA and EGTA increased the enzyme activity by 97 and 70%, respectively. The surfactant Triton X-100 reduced the enzyme activity by 50% and lowered the optimum temperature to 60°C. However, the thermostability of the enzyme PF2001Δ60 was enhanced with Triton X-100. PMID:21760993

  13. Neutron crystallographic study on rubredoxin from Pyrococcus furiosus by BIX-3, a single-crystal diffractometer for biomacromolecules.

    PubMed

    Kurihara, Kazuo; Tanaka, Ichiro; Chatake, Toshiyuki; Adams, Michael W W; Jenney, Francis E; Moiseeva, Natalia; Bau, Robert; Niimura, Nobuo

    2004-08-01

    The structure of a partially deuterated rubredoxin from the hyperthermophilic archaeon Pyrococcus furiosus, an organism that grows optimally at 100 degrees C, was determined by using the neutron single-crystal diffractometer dedicated for biological macromolecules (BIX-3) at the JRR-3M reactor of the Japan Atomic Energy Research Institute. Data were collected at room temperature up to a resolution of 1.5 A, and the completeness factor of the data set was 81.9%. The model contains 306 H and 50 D atoms. A total of 37 hydration water molecules were identified, with 15 having all three atoms fully located and the remaining D2O molecules partially defined. The model has been refined to final agreement factors of R = 18.6% and Rfree = 21.7%. Several orientations of the O-D bonds of side chains, whose assignments from x-ray data were previously ambiguous, were clearly visible in the neutron structure. Although most backbone N-H bonds had undergone some degree of H/D exchange throughout the rubredoxin molecule, 5 H atom positions still had distinctly negative (H) peaks. The neutron Fourier maps clearly showed the details of an extensive set of H bonds involving the ND3+ terminus that may contribute to the unusual thermostability of this molecule. PMID:15272083

  14. An In Silico Approach for Characterization of an Aminoglycoside Antibiotic-Resistant Methyltransferase Protein from Pyrococcus furiosus (DSM 3638)

    PubMed Central

    Oany, Arafat Rahman; Jyoti, Tahmina Pervin; Ahmad, Shah Adil Ishtiyaq

    2014-01-01

    Pyrococcus furiosus is a hyperthermophilic archaea. A hypothetical protein of this archaea, PF0847, was selected for computational analysis. Basic local alignment search tool and multiple sequence alignment (MSA) tool were employed to search for related proteins. Both the secondary and tertiary structure prediction were obtained for further analysis. Three-dimensional model was assessed by PROCHECK and QMEAN6 programs. To get insights about the physical and functional associations of the protein, STRING network analysis was performed. Binding of the SAM (S-adenosyl-l-methionine) ligand with our protein, fetched from an antibiotic-related methyltransferase (PDB code: 3P2K: D), showed high docking energy and suggested the function of the protein as methyltransferase. Finally, we tried to look for a specific function of the proposed methyltransferase, and binding of the geneticin bound to the eubacterial 16S rRNA A-site (PDB code: 1MWL) in the active site of the PF0847 gave us the indication to predict the protein responsible for aminoglycoside antibiotic resistance. PMID:24683305

  15. 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

  16. 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

  17. 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.

  18. 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

  19. Purification and Characterization of Two Functional Forms of Intracellular Protease PfpI from the Hyperthermophilic Archaeon Pyrococcus furiosus

    PubMed Central

    Halio, S. B.; Bauer, M. W.; Mukund, S.; Adams, M.; Kelly, R. M.

    1997-01-01

    The hyperthermophilic archaeon Pyrococcus furiosus grows optimally at 100(deg)C by the fermentation of peptides and carbohydrates. From this organism, we have purified to homogeneity an intracellular protease, previously designated PfpI (P. furiosus protease I) (S. B. Halio, I. I. Blumentals, S. A. Short, B. M. Merrill, and R. M. Kelly, J. Bacteriol. 178:2605-2612, 1996). The protease contains a single subunit with a molecular mass of approximately 19 kDa and exists in at least two functional conformations, which were purified separately. The predominant form from the purification (designated PfpI-C1) is a hexamer with a molecular mass of 124 (plusmn) 6 kDa (by gel filtration) and comprises about 90% of the total activity. The minor form (designated PfpI-C2) is trimeric with a molecular mass of 59 (plusmn) 3 kDa. PfpI-C1 hydrolyzed both basic and hydrophobic residues in the P1 position, indicating trypsin- and chymotrypsin-like specificities, respectively. The temperature optimum for Ala-Ala-Phe-7-amido-4-methylcoumarin (AAF-MCA) hydrolysis was (symbl)85(deg)C both for purified PfpI-C1 and for proteolytic activity in P. furiosus cell extract. In contrast, the temperature optimum for PfpI prepared by incubating a cell extract of P. furiosus at 98(deg)C in 1% sodium dodecyl sulfate for 24 h at 95 to 100(deg)C (I. I. Blumentals, A. S. Robinson, and R. M. Kelly, Appl. Environ. Microbiol. 56:1255-1262, 1990), designated PfpI-H, was (symbl)100(deg)C. Moreover, the half-life of activity of PfpI-C1 at 98(deg)C was less than 30 min, in contrast to a value of more than 33 h measured for PfpI-H. PfpI-C1 appears to be a predominant serine-type protease in cell extracts but is converted in vitro, probably in part by deamidation of Asn and Gln residues, to a more thermally stable form (PfpI-H) by prolonged heat treatment. The deamination hypothesis is supported by the differences in the measured pI values of PfpI-C1 (6.1) and PfpI-H (3.8). High levels of potassium phosphate (>0

  20. 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

  1. 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

  2. Improving dideoxynucleotide-triphosphate utilisation by the hyper-thermophilic DNA polymerase from the archaeon Pyrococcus furiosus

    PubMed Central

    Evans, Steven J.; Fogg, Mark J.; Mamone, Anthony; Davis, Maria; Pearl, Laurence H.; Connolly, Bernard A.

    2000-01-01

    Polymerases from the Pol-I family which are able to efficiently use ddNTPs have demonstrated a much improved performance when used to sequence DNA. A number of mutations have been made to the gene coding for the Pol-II family DNA polymerase from the archaeon Pyrococcus furiosus with the aim of improving ddNTP utilisation. ‘Rational’ alterations to amino acids likely to be near the dNTP binding site (based on sequence homologies and structural information) did not yield the desired level of selectivity for ddNTPs. However, alteration at four positions (Q472, A486, L490 and Y497) gave rise to variants which incorporated ddNTPs better than the wild type, allowing sequencing reactions to be carried out at lowered ddNTP:dNTP ratios. Wild-type Pfu–Pol required a ddNTP:dNTP ratio of 30:1; values of 5:1 (Q472H), 1:3 (L490W), 1:5 (A486Y) and 5:1 (Y497A) were found with the four mutants; A486Y representing a 150-fold improvement over the wild type. A486, L490 and Y497 are on an α-helix that lines the dNTP binding groove, but the side chains of the three amino acids point away from this groove; Q472 is in a loop that connects this α-helix to a second long helix. None of the four amino acids can contact the dNTP directly. Therefore, the increased selectivity for ddNTPs is likely to arise from two factors: (i) small overall changes in conformation that subtly alter the nucleotide triphosphate binding site such that ddNTPs become favoured; (ii) interference with a conformational change that may be critical both for the polymerisation step and discrimination between different nucleotide triphosphates. PMID:10666444

  3. A billion-fold range in acidity for the solvent-exposed amides of Pyrococcus furiosus rubredoxin.

    PubMed

    Anderson, Janet S; Hernández, Griselda; Lemaster, David M

    2008-06-10

    The exchange rates of the static solvent-accessible amide hydrogens of Pyrococcus furiosus rubredoxin range from near the diffusion-limited rate to a billion-fold slower for the non-hydrogen-bonded Val 38 (eubacterial numbering). Hydrogen exchange directly monitors the kinetic acidity of the peptide nitrogen. Electrostatic solvation free energies were calculated by Poisson-Boltzmann methods for the individual peptide anions that form during the hydroxide-catalyzed exchange reaction to examine how well the predicted thermodynamic acidities match the experimentally determined kinetic acidities. With the exception of the Ile 12 amide, the differential exchange rate constant for each solvent-exposed amide proton that is not hydrogen bonded to a backbone carbonyl can be predicted within a factor of 6 (10 (0.78)) root-mean-square deviation (rmsd) using the CHARMM22 electrostatic parameter set and an internal dielectric value of 3. Under equivalent conditions, the PARSE parameter set yields a larger rmsd value of 1.28 pH units, while the AMBER parm99 parameter set resulted in a considerably poorer correlation. Either increasing the internal dielectric value to 4 or reducing it to a value of 2 significantly degrades the quality of the prediction. Assigning the excess charge of the peptide anion equally between the peptide nitrogen and the carbonyl oxygen also reduces the correlation to the experimental data. These continuum electrostatic calculations were further analyzed to characterize the specific structural elements that appear to be responsible for the wide range of peptide acidities observed for these solvent-exposed amides. The striking heterogeneity in the potential at sites along the protein-solvent interface should prove germane to the ongoing challenge of quantifying the contribution that electrostatic interactions make to the catalytic acceleration achieved by enzymes. PMID:18479148

  4. 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

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

    PubMed Central

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

    2014-01-01

    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 Mg2+-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. PMID:25361963

  6. 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

  7. 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

  8. Expression, purification, crystallization and preliminary crystallographic analysis of a stand-alone RAM domain with hydrolytic activity from the hyperthermophile Pyrococcus furiosus

    SciTech Connect

    Agapay, Ramelito C.; Savvides, Savvas N.; Van Driessche, Gonzalez; Devreese, Bart; Van Beeumen, Jozef; Jongejan, Jaap A. Hagen, Wilfred R.

    2005-10-01

    A P. furiosus stand-alone RAM domain with hydrolytic activity has been cloned and expressed in E. coli. The purified protein was crystallized alone and with EPNP and PMSF, producing crystals that yield diffraction data to resolutions of 2.8, 2.2 and 2.8 Å, respectively. The RAM domain is one of several ligand-binding modules present in prokaryotes that are presumed to regulate the transcription of specific genes. To date, no hydrolytic activity has been reported for such modules. Curiously, a stand-alone RAM domain in Pyrococcus furiosus was isolated during a screen for hydrolytic activity against chromogenic esters. The gene encoding this protein was cloned and expressed in Escherichia coli and crystallized after a single purification step. X-ray diffraction data from the crystals were obtained to a resolution of 2.8 Å using a conventional X-ray source. The cocrystallization of the recombinant protein with 1,2-epoxy-3-(4-nitrophenoxy)propane (EPNP) and phenylmethylsulfonyl fluoride (PMSF) produced crystals that yielded data to 2.2 and 2.8 Å, respectively, using synchrotron radiation. Both the untreated and EPNP-treated crystals crystallize isomorphously in space group C2 and contain three dimers in the asymmetric unit. The PMSF-treated crystals also belong to this space group and have almost identical packing density, but show dramatically different unit-cell parameters.

  9. Hydrolysis of flavanone glycosides by β-glucosidase from Pyrococcus furiosus and its application to the production of flavanone aglycones from citrus extracts.

    PubMed

    Shin, Kyung-Chul; Nam, Hyun-Koo; Oh, Deok-Kun

    2013-11-27

    The hydrolytic activity of the recombinant β-glucosidase from Pyrococcus furiosus for the flavanone glycoside hesperidin was optimal at pH 5.5 and 95 °C in the presence of 0.5% (v/v) dimethyl sulfoxide (DMSO) and 0.1% (w/v) Tween 40 with a half-life of 88 h, a Km of 1.6 mM, and a kcat of 68.4 1/s. The specific activity of the enzyme for flavonoid glycosides followed the order hesperidin > neohesperidin > naringin > narirutin > poncirin > diosmin > neoponcirin > rutin. The specific activity for flavanone was higher than that for flavone or flavonol. DMSO at 10% (v/v) was used to increase the solubility of flavanone glycosides as substrates. The enzyme completely converted flavanone glycosides (1 g/L) to flavanone aglycones and disaccharides via one-step reaction. The major flavanone in grapefruit peel, grapefruit pulp, or orange peel extract was naringin (47.5 mg/g), naringin (16.6 mg/g), or hesperidin (18.2 mg/g), respectively. β-Glucosidase from P. furiosus completely converted naringin and narirutin in 100% (w/v) grapefruit peel extract to 22.5 g/L naringenin after 12 h, with a productivity of 1.88 g L(-1) h(-1); naringin and narirutin in 100% (w/v) grapefruit pulp extract to 8.1 g/L naringenin after 9 h, with a productivity of 0.90 g L(-1) h(-1); and hesperidin in 100% (w/v) orange peel extract to 9.0 g/L hesperetin after 9 h, with a productivity of 1.00 g L(-1) h(-1). The conversion yields, concentrations, and productivities of flavanone aglycones in this study are the highest among those obtained from citrus extracts. Thus, this enzyme may be useful for the industrial hydrolysis of flavanone glycosides in citrus extracts. PMID:24188428

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

    PubMed

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

    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 (S0) to hydrogen sulfide (H2S) for optimal growth, even though S0 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 H2S if S0 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 S0 and polysulfide served as substrates for H2S production, and the S0 reduction activity but not the H2-oxidation activity was enhanced by the redox protein rubredoxin. The H2-oxidizing and S0-reduction activities of the enzyme also showed different responses to pH, temperature, and inhibitors. This bifunctional "sulfhydrogenase" 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 S0 to H2S. It is suggested that the function of some form of ancestral hydrogenase was S0 reduction rather than, or in addition to, the reduction of protons. PMID:8389482

  11. Solution structure of Pyrococcus furiosus RPP21, a component of the archaeal RNase P holoenzyme, and interactions with its RPP29 protein partner.

    PubMed

    Amero, Carlos D; Boomershine, William P; Xu, Yiren; Foster, Mark

    2008-11-11

    RNase P is the ubiquitous ribonucleoprotein metalloenzyme responsible for cleaving the 5'-leader sequence of precursor tRNAs during their maturation. While the RNA subunit is catalytically active on its own at high monovalent and divalent ion concentrations, four protein subunits are associated with archaeal RNase P activity in vivo: RPP21, RPP29, RPP30, and POP5. These proteins have been shown to function in pairs: RPP21-RPP29 and POP5-RPP30. We have determined the solution structure of RPP21 from the hyperthermophilic archaeon Pyrococcus furiosus ( Pfu) using conventional and paramagnetic NMR techniques. Pfu RPP21 in solution consists of an unstructured N-terminus, two alpha-helices, a zinc binding motif, and an unstructured C-terminus. Moreover, we have used chemical shift perturbations to characterize the interaction of RPP21 with RPP29. The data show that the primary contact with RPP29 is localized to the two helices of RPP21. This information represents a fundamental step toward understanding structure-function relationships of the archaeal RNase P holoenzyme. PMID:18922021

  12. Stoichiometric complex formation by proliferating cell nuclear antigen (PCNA) and its interacting protein: purification and crystallization of the DNA polymerase and PCNA monomer mutant complex from Pyrococcus furiosus

    SciTech Connect

    Nishida, Hirokazu; Matsumiya, Shigeki; Tsuchiya, Daisuke; Ishino, Yoshizumi; Morikawa, Kosuke

    2006-03-01

    A stable stoichiometric complex of archaeal DNA polymerase with proliferating cell nuclear antigen (PCNA) was formed using a PCNA monomer mutant and the complex was successfully crystallized. Replicative DNA polymerase interacts with processivity factors, the β-subunit of DNA polymerase III or proliferating cell nuclear antigen (PCNA), in order to function with a long template DNA. The archaeal replicative DNA polymerase from Pyrococcus furiosus interacts with PCNA via its PCNA-interacting protein (PIP) motif at the C-terminus. The PCNA homotrimeric ring contains one PIP interacting site on each monomer and since the ring can accommodate up to three molecules simultaneously, formation of a stable stoichiometric complex of PCNA with its interacting protein has been difficult to control in vitro. A stable complex of the DNA polymerase with PCNA, using a PCNA monomer mutant, has been purified and crystallized. The best ordered crystal diffracted to 3.0 Å resolution using synchrotron radiation. The crystals belong to space group P2{sub 1}2{sub 1}2, with unit-cell parameters a = 225.3, b = 123.3, c = 91.3 Å.

  13. 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

  14. '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

  15. 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.

  16. 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. PMID:17204331

  17. 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.

  18. 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

  19. 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

  20. 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

  1. Solution Structure of an Archaeal RNase P Binary Protein Complex. Formation of the 30-kDa Complex Between Pyrococcus furiosus RPP21 and RPP29 is Accompanied by Coupled Protein Folding, and Highlights Critical Features for Protein-Protein and Protein-RNA Interactions

    PubMed Central

    Xu, Yiren; Amero, Carlos D.; Pulukkunat, Dileep K.; Gopalan, Venkat; Foster, Mark P.

    2009-01-01

    RNase P is a ribonucleoprotein (RNP) enzyme that catalyzes the Mg2+-dependent 5’ maturation of precursor tRNAs. In all domains of life, it is a ribozyme: the RNase P RNA (RPR) component has been demonstrated to be responsible for catalysis. However, the number of RNase P protein subunits (RPPs) varies from one in bacteria to nine or ten in eukarya. The archaeal RPR is associated with at least four RPPs, which function in pairs (RPP21–RPP29 and RPP30-POP5). We used solution NMR spectroscopy to determine the three-dimensional structure of the protein-protein complex comprising Pyrococcus furiosus (Pfu) RPP21 and RPP29. We found that the protein-protein interaction is characterized by coupled folding of secondary structural elements that participate in interface formation. In addition to detailing the intermolecular contacts that stabilize this 30-kDa binary complex, the structure identifies surfaces rich in conserved basic residues likely vital for recognition of the RPR and/or precursor tRNA. Furthermore, enzymatic footprinting experiments allowed us to localize the RPP21–RPP29 complex to the specificity domain of the RPR. These findings provide valuable new insights into mechanisms of RNP assembly and serve as important steps towards a three-dimensional model of this ancient RNP enzyme. PMID:19733182

  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. Improving the catalytic activity of hyperthermophilic Pyrococcus prolidases for detoxification of organophosphorus nerve agents over a broad range of temperatures.

    PubMed

    Theriot, Casey M; Du, Xuelian; Tove, Sherry R; Grunden, Amy M

    2010-08-01

    Prolidase isolated from the hyperthermophilic archaeon Pyrococcus furiosus has potential for application for decontamination of organophosphorus compounds in certain pesticides and chemical warfare agents under harsh conditions. However, current applications that use an enzyme-based cocktail are limited by poor long-term enzyme stability and low reactivity over a broad range of temperatures. To obtain a better enzyme for OP nerve agent decontamination and to investigate structural factors that influence protein thermostability and thermoactivity, randomly mutated P. furiosus prolidases were prepared by using XL1-red-based mutagenesis and error-prone PCR. An Escherichia coli strain JD1 (lambdaDE3) (auxotrophic for proline [DeltaproA] and having deletions in pepQ and pepP dipeptidases with specificity for proline-containing dipeptides) was constructed for screening mutant P. furiosus prolidase expression plasmids. JD1 (lambdaDE3) cells were transformed with mutated prolidase expression plasmids and plated on minimal media supplemented with 50 muM Leu-Pro as the only source of proline. By using this positive selection, Pyrococcus prolidase mutants with improved activity over a broader range of temperatures were isolated. The activities of the mutants over a broad temperature range were measured for both Xaa-Pro dipeptides and OP nerve agents, and the thermoactivity and thermostability of the mutants were determined. PMID:20422176

  4. Co-expression of chaperones from P. furiosus enhanced the soluble expression of the recombinant hyperthermophilic α-amylase in E. coli.

    PubMed

    Peng, Shuaiying; Chu, Zhongmei; Lu, Jianfeng; Li, Dongxiao; Wang, Yonghong; Yang, Shengli; Zhang, Yi

    2016-05-01

    The extracellular α-amylase from the hyperthermophilic archaeum Pyrococcus furiosus (PFA) is extremely thermostable and of an industrial importance and interest. PFA aggregates and accumulates as insoluble inclusion bodies when expressed as a heterologous protein at a high level in Escherichia coli. In the present study, we investigated the roles of chaperones from P. furiosus in the soluble expression of recombinant PFA in E. coli. The results indicate that co-expression of PFA with the molecular chaperone prefoldin alone significantly increased the soluble expression of PFA. Although, co-expression of other main chaperone components from P. furiosus, such as the small heat shock protein (sHSP) or chaperonin (HSP60), was also able to improve the soluble expression of PFA to a certain extent. Co-expression of chaperonin or sHSP in addition to prefoldin did not further increase the soluble expression of PFA. This finding emphasizes the biotechnological potentials of the molecular chaperone prefoldin from P. furiosus, which may facilitate the production of recombinant PFA. PMID:26862080

  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. 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. PMID:26337425

  8. 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

  9. Improving the catalytic activity of hyperthermophilic Pyrococcus horikoshii prolidase for detoxification of organophosphorus nerve agents over a broad range of temperatures.

    PubMed

    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. Cell-free transcription at 95 degrees: thermostability of transcriptional components and DNA topology requirements of Pyrococcus transcription.

    PubMed Central

    Hethke, C; Bergerat, A; Hausner, W; Forterre, P; Thomm, M

    1999-01-01

    Cell-free transcription of archaeal promoters is mediated by two archaeal transcription factors, aTBP and TFB, which are orthologues of the eukaryotic transcription factors TBP and TFIIB. Using the cell-free transcription system described for the hyperthermophilic Archaeon Pyrococcus furiosus by Hethke et al., the temperature limits and template topology requirements of archaeal transcription were investigated. aTBP activity was not affected after incubation for 1 hr at 100 degrees. In contrast, the half-life of RNA polymerase activity was 23 min and that of TFB activity was 3 min. The half-life of a 328-nt RNA product was 10 min at 100 degrees. Best stability of RNA was observed at pH 6, at 400 mm K-glutamate in the absence of Mg(2+) ions. Physiological concentrations of K-glutamate were found to stabilize protein components in addition, indicating that salt is an important extrinsic factor contributing to thermostability. Both RNA and proteins were stabilized by the osmolyte betaine at a concentration of 1 m. The highest activity for RNA synthesis at 95 degrees was obtained in the presence of 1 m betaine and 400 mm K-glutamate. Positively supercoiled DNA, which was found to exist in Pyrococcus cells, can be transcribed in vitro both at 70 degrees and 90 degrees. However, negatively supercoiled DNA was the preferred template at all temperatures tested. Analyses of transcripts from plasmid topoisomers harboring the glutamate dehydrogenase promoter and of transcription reactions conducted in the presence of reverse gyrase indicate that positive supercoiling of DNA inhibits transcription from this promoter. PMID:10430563

  11. Characterization of recombinant glutamine synthetase from the hyperthermophilic archaeon Pyrococcus sp. strain KOD1.

    PubMed Central

    Adul Rahman, R N; Jongsareejit, B; Fujiwara, S; Imanaka, T

    1997-01-01

    The glnA gene encoding glutamine synthetase was cloned from the hyperthermophilic archaeon Pyrococcus sp. strain KOD1, and its nucleotide sequence was determined. The glnA gene was expressed in Escherichia coli ME8459 (glnA mutant strain), and the protein was purified to homogeneity and shown to be functional in a dodecameric from (637,000 Da), exhibiting both transferase and synthetase activities. However, kinetic studies indicated that the enzyme possessed low biosynthetic activity, suggesting that the reaction was biased towards glutamate production. The optimum temperature for both activities was 60 degrees C, which was lower than the optimal growth temperature of KOD1. Recombinant KOD1 GlnA exhibited different optimum pHs depending on the reaction employed (pH 7.8 for the synthetase reaction and pH 7.2 for the transferase reaction). Of the various nucleoside triphosphates tested, GTP as well as ATP was involved in the synthetase reaction. PMID:9172372

  12. A Self-compartmentalizing Hexamer Serine Protease from Pyrococcus Horikoshii

    PubMed Central

    Menyhárd, Dóra K.; Kiss-Szemán, Anna; Tichy-Rács, Éva; Hornung, Balázs; Rádi, Krisztina; Szeltner, Zoltán; Domokos, Klarissza; Szamosi, Ilona; Náray-Szabó, Gábor; Polgár, László; Harmat, Veronika

    2013-01-01

    Oligopeptidases impose a size limitation on their substrates, the mechanism of which has long been under debate. Here we present the structure of a hexameric serine protease, an oligopeptidase from Pyrococcus horikoshii (PhAAP), revealing a complex, self-compartmentalized inner space, where substrates may access the monomer active sites passing through a double-gated “check-in” system, first passing through a pore on the hexamer surface and then turning to enter through an even smaller opening at the monomers' domain interface. This substrate screening strategy is unique within the family. We found that among oligopeptidases, a residue of the catalytic apparatus is positioned near an amylogenic β-edge, which needs to be protected to prevent aggregation, and we found that different oligopeptidases use different strategies to achieve such an end. We propose that self-assembly within the family results in characteristically different substrate selection mechanisms coupled to different multimerization states. PMID:23632025

  13. 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.

  14. 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.

  15. The key to the extraordinary thermal stability of P. furiosus holo-rubredoxin: iron binding-guided packing of a core aromatic cluster responsible for high kinetic stability of the native structure.

    PubMed

    Prakash, Satya; Sundd, Monica; Guptasarma, Purnananda

    2014-01-01

    Pyrococcus furiosus rubredoxin (PfRd), a small, monomeric, 53 residues-long, iron-containing, electron-transfer protein of known structure is sometimes referred to as being the most structurally-stable protein known to man. Here, using a combination of mutational and spectroscopic (CD, fluorescence, and NMR) studies of differently made holo- and apo-forms of PfRd, we demonstrate that it is not the presence of iron, or even the folding of the PfRd chain into a compact well-folded structure that causes holo-PfRd to display its extraordinary thermal stability, but rather the correct iron binding-guided packing of certain residues (specifically, Trp3, Phe29, Trp36, and also Tyr10) within a tight aromatic cluster of six residues in PfRd's hydrophobic core. Binding of the iron atom appears to play a remarkable role in determining subtle details of residue packing, forcing the chain to form a hyper-thermally stable native structure which is kinetically stable enough to survive (subsequent) removal of iron. On the other hand, failure to bind iron causes the same chain to adopt an equally well-folded native-like structure which, however, has a differently-packed aromatic cluster in its core, causing it to be only as stable as any other ordinary mesophile-derived rubredoxin. Our studies demonstrate, perhaps for the very first time ever that hyperthermal stability in proteins can owe to subtle differences in residue packing vis a vis mesostable proteins, without there being any underlying differences in either amino acid sequence, or bound ligand status. PMID:24603898

  16. Analysis of the complete genome sequence of the archaeon Pyrococcus chitonophagus DSM 10152 (formerly Thermococcus chitonophagus).

    PubMed

    Papadimitriou, Konstantinos; Baharidis, Panagiotis K; Georgoulis, Anastasios; Engel, Marion; Louka, Maria; Karamolegkou, Georgia; Tsoka, Aggeliki; Blom, Jochen; Pot, Bruno; Malecki, Piotr; Rypniewski, Wojciech; Huber, Harald; Schloter, Michael; Vorgias, Constantinos

    2016-05-01

    Here we analyze the first complete genome sequence of Pyrococcus chitonophagus. The archaeon was previously suggested to belong to the Thermococcus rather than the Pyrococcus genus. Whole genome phylogeny as well as whole proteome comparisons using all available complete genomes in Thermococcales clearly showed that the species belongs to the Pyrococcus genus. P. chitonophagus was originally isolated from a hydrothermal vent site and it has been described to effectively degrade chitin debris, and therefore is considered to play a major role in the sea water ecology and metabolic activity of microbial consortia within hot sea water ecosystems. Indeed, an obvious feature of the P. chitonophagus genome is that it carries proteins showing complementary activities for chitin degradation, i.e. endo- and exo-chitinase, diacetylchitobiose deacetylase and exo-β-D glucosaminidase activities. This finding supports the hypothesis that compared to other Thermococcales species P. chitonophagus is adapted to chitin degradation. PMID:27016195

  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. 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

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

    PubMed

    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. 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.

  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. Pyrococcus horikoshii TET2 Peptidase Assembling Process and Associated Functional Regulation*

    PubMed Central

    Appolaire, Alexandre; Rosenbaum, Eva; Durá, M. Asunción; Colombo, Matteo; Marty, Vincent; Savoye, Marjolaine Noirclerc; Godfroy, Anne; Schoehn, Guy; Girard, Eric; Gabel, Frank; Franzetti, Bruno

    2013-01-01

    Tetrahedral (TET) aminopeptidases are large polypeptide destruction machines present in prokaryotes and eukaryotes. Here, the rules governing their assembly into hollow 12-subunit tetrahedrons are addressed by using TET2 from Pyrococcus horikoshii (PhTET2) as a model. Point mutations allowed the capture of a stable, catalytically active precursor. Small angle x-ray scattering revealed that it is a dimer whose architecture in solution is identical to that determined by x-ray crystallography within the fully assembled TET particle. Small angle x-ray scattering also showed that the reconstituted PhTET2 dodecameric particle displayed the same quaternary structure and thermal stability as the wild-type complex. The PhTET2 assembly intermediates were characterized by analytical ultracentrifugation, native gel electrophoresis, and electron microscopy. They revealed that PhTET2 assembling is a highly ordered process in which hexamers represent the main intermediate. Peptide degradation assays demonstrated that oligomerization triggers the activity of the TET enzyme toward large polypeptidic substrates. Fractionation experiments in Pyrococcus and Halobacterium cells revealed that, in vivo, the dimeric precursor co-exists together with assembled TET complexes. Taken together, our observations explain the biological significance of TET oligomerization and suggest the existence of a functional regulation of the dimer-dodecamer equilibrium in vivo. PMID:23696647

  4. Molecular characterization of the group II chaperonin from the hyperthermophilic archaeum Pyrococcus horikoshii OT3.

    PubMed

    Okochi, Mina; Matsuzaki, Hiroki; Nomura, Tomoko; Ishii, Noriyuki; Yohda, Masafumi

    2005-04-01

    The group II chaperonin from the hyperthermophilic archaeum Pyrococcus horikoshii OT3 (PhCPN) and its functional cooperation with the cognate prefoldin were investigated. PhCPN existed as a homo-oligomer in a double-ring structure, which protected the citrate synthase of a porcine heart from thermal aggregation at 45 degrees C, and did the same on the isopropylmalate dehydrogenase (IPMDH) of a thermophilic bacterium, Thermus thermophilus HB8, at 90 degrees C. PhCPN also enhanced the refolding of green fluorescent protein (GFP), which had been unfolded by low pH, in an ATP-dependent manner. Unexpectedly, functional cooperation between PhCPN and Pyrococcus prefoldin (PhPFD) in the refolding of GFP was not observed. Instead, cooperation between PhCPN and PhPFD was observed in the refolding of IPMDH unfolded with guanidine hydrochloride. Although PhCPN alone was not effective in the refolding of IPMDH, the refolding efficiency was enhanced by the cooperation of PhCPN with PhPFD. PMID:15538645

  5. Glyceraldehyde-3-phosphate dehydrogenase from the hyperthermophilic archaebacterium Pyrococcus woesei: characterization of the enzyme, cloning and sequencing of the gene, and expression in Escherichia coli.

    PubMed Central

    Zwickl, P; Fabry, S; Bogedain, C; Haas, A; Hensel, R

    1990-01-01

    The glyceraldehyde-3-phosphate dehydrogenase from the hyperthermophilic archaebacterium Pyrococcus woesei (optimal growth temperature, 100 to 103 degrees C) was purified to homogeneity. This enzyme was strictly phosphate dependent, utilized either NAD+ or NADP+, and was insensitive to pentalenolactone like the enzyme from the methanogenic archaebacterium Methanothermus fervidus. The enzyme exhibited a considerable thermostability, with a 44-min half-life at 100 degrees C. The amino acid sequence of the glyceraldehyde-3-phosphate dehydrogenase from P. woesei was deduced from the nucleotide sequence of the coding gene. Compared with the enzyme homologs from mesophilic archaebacteria (Methanobacterium bryantii, Methanobacterium formicicum) and an extremely thermophilic archaebacterium (Methanothermus fervidus), the primary structure of the P. woesei enzyme exhibited a strikingly high proportion of aromatic amino acid residues and a low proportion of sulfur-containing residues. The coding gene of P. woesei was expressed at a high level in Escherichia coli, thus providing an ideal basis for detailed structural and functional studies of that enzyme. Images PMID:2165475

  6. 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

  7. Cloning, Expression, and Purification of Hyperthermophile α-Amylase from Pyrococcus woesei

    PubMed Central

    Ghasemi, Amir; Ghafourian, Sobhan; Vafaei, Sedighe; Mohebi, Reza; Farzi, Maryam; Taherikalani, Morovat; Sadeghifard, Nourkhoda

    2015-01-01

    Objectives In an attempt α-amylase gene from Pyrococcus woesei was amplified and cloned into a pTYB2 vector to generate the recombinant plasmid pTY- α-amylase. Methods Escherichia coli BL21 used as a host and protein expression was applied using IPTG. SDS-PAGE assay demonstrated the 100 kDa protein. Amylolytic activity of proteins produced by transformed E. coli cells was detected by zymography, and the rate of active α-amylase with and without the intein tag in both soluble conditions and as inclusion bodies solubilized by 4M urea were measured. Results Amylolytic activity of ∼185,000 U/L of bacterial culture was observed from the soluble form of the protein using this system. Conclusion These results indicate that this expression system was appropriate for the production of thermostable α-amylase. PMID:26835242

  8. 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.

  9. A self-compartmentalizing hexamer serine protease from Pyrococcus horikoshii: substrate selection achieved through multimerization.

    PubMed

    Menyhárd, Dóra K; Kiss-Szemán, Anna; Tichy-Rács, Éva; Hornung, Balázs; Rádi, Krisztina; Szeltner, Zoltán; Domokos, Klarissza; Szamosi, Ilona; Náray-Szabó, Gábor; Polgár, László; Harmat, Veronika

    2013-06-14

    Oligopeptidases impose a size limitation on their substrates, the mechanism of which has long been under debate. Here we present the structure of a hexameric serine protease, an oligopeptidase from Pyrococcus horikoshii (PhAAP), revealing a complex, self-compartmentalized inner space, where substrates may access the monomer active sites passing through a double-gated "check-in" system, first passing through a pore on the hexamer surface and then turning to enter through an even smaller opening at the monomers' domain interface. This substrate screening strategy is unique within the family. We found that among oligopeptidases, a residue of the catalytic apparatus is positioned near an amylogenic β-edge, which needs to be protected to prevent aggregation, and we found that different oligopeptidases use different strategies to achieve such an end. We propose that self-assembly within the family results in characteristically different substrate selection mechanisms coupled to different multimerization states. PMID:23632025

  10. Isolation and Characterization of a Second Subunit of Molecular Chaperonin from Pyrococcus kodakaraensis KOD1: Analysis of an ATPase-Deficient Mutant Enzyme

    PubMed Central

    Izumi, Michi; Fujiwara, Shinsuke; Takagi, Masahiro; Kanaya, Shigenori; Imanaka, Tadayuki

    1999-01-01

    The cpkA gene encoding a second (α) subunit of archaeal chaperonin from Pyrococcus kodakaraensis KOD1 was cloned, sequenced, and expressed in Escherichia coli. Recombinant CpkA was studied for chaperonin functions in comparison with CpkB (β subunit). The effect on decreasing the insoluble form of proteins was examined by coexpressing CpkA or CpkB with CobQ (cobyric acid synthase from P. kodakaraensis) in E. coli. The results indicate that both CpkA and CpkB effectively decrease the amount of the insoluble form of CobQ. Both CpkA and CpkB possessed the same ATPase activity as other bacterial and eukaryal chaperonins. The ATPase-deficient mutant proteins CpkA-D95K and CpkB-D95K were constructed by changing conserved Asp95 to Lys. Effect of the mutation on the ATPase activity and CobQ solubilization was examined. Neither mutant exhibited ATPase activity in vitro. Nevertheless, they decreased the amount of the insoluble form of CobQ by coexpression as did wild-type CpkA and CpkB. These results implied that both CpkA and CpkB could assist protein folding for nascent protein in E. coli without requiring energy from ATP hydrolysis. PMID:10103287

  11. Identification and characterization of a thermostable bifunctional enzyme with phosphomannose isomerase and sugar-1-phosphate nucleotidylyltransferase activities from a hyperthermophilic archaeon, Pyrococcus horikoshii OT3.

    PubMed

    Akutsu, Jun-ichi; Zhang, Zilian; Morita, Rihito; Kawarabayasi, Yutaka

    2015-11-01

    Mannosylglycerate is known as a compatible solute, and plays important roles for salinity adaptation and high temperature stability of microorganisms. In the gene cluster for the mannosylglycerate biosynthetic pathway predicted from the genomic data of Pyrococcus horikoshii OT3, the PH0925 protein was found as a putative bifunctional enzyme with phosphomannose isomerase (PMI) and mannose-1-phosphate guanylyltransferase (Man-1-P GTase) activities, which can synthesize GDP-mannose when accompanied by a phosphomannomutase/phosphoglucomutase (PMM/PGM) enzyme (PH0923). The recombinant PH0925 protein, expressed in E. coli, exhibited both expected PMI and Man-1-P GTase activities, as well as absolute thermostability; 95 °C was the optimum reaction temperature. According to the guanylyltransferase activity (GTase) of the PH0925 protein, it was found that the protein can catalyze glucose-1-phosphate (Glc-1-P) and glucosamine-1-phosphate (GlcN-1-P) in addition to Man-1-P. The analyses of C-terminus-truncated forms of the PH0925 protein indicated that sugar-1-phosphate nucleotidylyltransferase (Sugar-1-P NTase) activity was located in the region from the N-terminus to the 345th residue, and that the C-terminal 114 residue region of the PH0925 protein inhibited the Man-1-P GTase activity. Conversely, the PMI activity was abolished by deletion of the C-terminal 14 residues. This is the first report of a thermostable enzyme with both PMI and multiple Sugar-1-P NTase activities. PMID:26290359

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

    PubMed

    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

  13. 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

  14. 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

  15. 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

  16. Identification and characterization of an archaeal kojibiose catabolic pathway in the hyperthermophilic Pyrococcus sp. strain ST04.

    PubMed

    Jung, Jong-Hyun; Seo, Dong-Ho; Holden, James F; Park, Cheon-Seok

    2014-03-01

    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

  17. Insights into the hyperthermostability and unusual region-specificity of archaeal Pyrococcus abyssi tRNA m1A57/58 methyltransferase

    PubMed Central

    Guelorget, Amandine; Roovers, Martine; Guérineau, Vincent; Barbey, Carole; Li, Xuan; Golinelli-Pimpaneau, Béatrice

    2010-01-01

    The S-adenosyl-l-methionine dependent methylation of adenine 58 in the T-loop of tRNAs is essential for cell growth in yeast or for adaptation to high temperatures in thermophilic organisms. In contrast to bacterial and eukaryotic tRNA m1A58 methyltransferases that are site-specific, the homologous archaeal enzyme from Pyrococcus abyssi catalyzes the formation of m1A also at the adjacent position 57, m1A57 being a precursor of 1-methylinosine. We report here the crystal structure of P. abyssi tRNA m1A57/58 methyltransferase (PabTrmI), in complex with S-adenosyl-l-methionine or S-adenosyl-l-homocysteine in three different space groups. The fold of the monomer and the tetrameric architecture are similar to those of the bacterial enzymes. However, the inter-monomer contacts exhibit unique features. In particular, four disulfide bonds contribute to the hyperthermostability of the archaeal enzyme since their mutation lowers the melting temperature by 16.5°C. His78 in conserved motif X, which is present only in TrmIs from the Thermococcocales order, lies near the active site and displays two alternative conformations. Mutagenesis indicates His78 is important for catalytic efficiency of PabTrmI. When A59 is absent in tRNAAsp, only A57 is modified. Identification of the methylated positions in tRNAAsp by mass spectrometry confirms that PabTrmI methylates the first adenine of an AA sequence. PMID:20483913

  18. 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

  19. Crystallization and preliminary X-ray analysis of a RecB-family nuclease from the archaeon Pyrococcus abyssi

    SciTech Connect

    Ren, Bin; Kuhn, Joëlle; Meslet-Cladiere, Laurence; Myllykallio, Hannu; Ladenstein, Rudolf

    2007-05-01

    A RecB-like nuclease from the archaeon Pyrococcus abyssi was expressed, purified and crystallized. The crystals belong to the orthorhombic space group C222{sub 1} with a = 81.5, b = 159.8, c = 100.8 Å, and a native data set was collected to 2.65 Å resolution. Nucleases are required to process and repair DNA damage in living cells. One of the best studied nucleases is the RecB protein, which functions in Escherichia coli as a component of the RecBCD enzyme complex that amends double-strand breaks in DNA. Although archaea do not contain the RecBCD complex, a RecB-like nuclease from Pyrococcus abyssi has been cloned, expressed and purified. The protein was crystallized by the sitting-drop vapour-diffusion method using polyethylene glycol 8000 as the precipitant. The crystals belong to the orthorhombic space group C222{sub 1}, with unit-cell parameters a = 81.5, b = 159.8, c = 100.8 Å. Self-rotation function and native Patterson map calculations revealed that there is a dimer in the asymmetric unit with its local twofold axis running parallel to the crystallographic twofold screw axis. The crystals diffracted to about 2 Å and a complete native data set was collected to 2.65 Å resolution.

  20. Cloning, expression, purification, crystallization and preliminary X-ray diffraction data of the Pyrococcus horikoshii RadA intein

    PubMed Central

    Lyskowski, Andrzej; Oeemig, Jesper S.; Jaakkonen, Anniina; Rommi, Katariina; DiMaio, Frank; Zhou, Dongwen; Kajander, Tommi; Baker, David; Wlodawer, Alexander; Goldman, Adrian; Iwaï, Hideo

    2011-01-01

    The RadA intein from the hyperthermophilic archaebacterium Pyrococcus horikoshii was cloned, expressed and purified for subsequent structure determination. The protein crystallized rapidly in several conditions. The best crystals, which diffracted to 1.75 Å resolution, were harvested from drops consisting of 0.1 M HEPES pH 7.5, 3.0 M NaCl and were cryoprotected with Paratone-N before flash-cooling. The collected data were processed in the orthorhombic space group P212121, with unit-cell parameters a = 58.1, b = 67.4, c = 82.9 Å. Molecular replacement with Rosetta using energy- and density-guided structure optimization provided the initial solution, which is currently under refinement. PMID:21543876

  1. 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…

  2. An Exhibit for Touching.

    ERIC Educational Resources Information Center

    Hunt, Susan

    1979-01-01

    An exhibit designed for visually handicapped persons presented by the Kalamazoo (Michigan) Institute of Art included bronze sculptures and oil paintings from the institute's permanent collection. (CL)

  3. San Rafael Schools Exhibit.

    ERIC Educational Resources Information Center

    San Rafael City Schools, CA.

    The San Rafael City Schools' exhibit which was displayed at the 1983 Marin County Fair (California) is described. The exhibit, entitled "Education - A Real Winner," consisted of 12 display panels illustrating the following aspects of the school system: (1) early history from 1861; (2) present board and administration; (3) present schools and…

  4. 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)

  5. Visitors Center Exhibits

    NASA Technical Reports Server (NTRS)

    1997-01-01

    A child enjoys building his own LEGO model at a play table which was included in the exhibit 'Travel in Space' World Show. The exhibit consisted of 21 displays designed to teach children about flight and space travel from the Wright brothers to future generations of space vehicles.

  6. 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

  7. 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.

  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. 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.

  10. 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

  11. 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.

  12. Overexpression, purification, crystallization and preliminary crystallographic studies of a hyperthermophilic adenylosuccinate synthetase from Pyrococcus horikoshii OT3

    PubMed Central

    Wang, Xiaoying; Akasaka, Ryogo; Takemoto, Chie; Morita, Satoshi; Yamaguchi, Machiko; Terada, Takaho; Shirozu, Mikako; Yokoyama, Shigeyuki; Chen, Shilin; Si, Shuyi; Xie, Yong

    2011-01-01

    Adenylosuccinate synthetase (AdSS) is a ubiquitous enzyme that catalyzes the first committed step in the conversion of inosine monophosphate (IMP) to adenosine monophosphate (AMP) in the purine-biosynthetic pathway. Although AdSS from the vast majority of organisms is 430–457 amino acids in length, AdSS sequences isolated from thermophilic archaea are 90–120 amino acids shorter. In this study, crystallographic studies of a short AdSS sequence from Pyrococcus horikoshii OT3 (PhAdSS) were performed in order to reveal the unusual structure of AdSS from thermophilic archaea. Crystals of PhAdSS were obtained by the microbatch-under-oil method and X-ray diffraction data were collected to 2.50 Å resolution. The crystal belonged to the trigonal space group P3212, with unit-cell parameters a = b = 57.2, c = 107.9 Å. There was one molecule per asymmetric unit, giving a Matthews coefficient of 2.17 Å3 Da−1 and an approximate solvent content of 43%. In contrast, the results of native polyacrylamide gel electrophoresis and analytical ultracentrifugation showed that the recombinant PhAdSS formed a dimer in solution. PMID:22139164

  13. 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.

  14. Purification, crystallization and preliminary crystallographic analysis of RecA superfamily ATPase PH0284 from Pyrococcus horikoshii OT3

    SciTech Connect

    Bagautdinov, Bagautdin; Kunishima, Naoki

    2006-04-01

    RecA superfamily ATPase PH0284 from P. horikoshii OT3 was overexpressed, purified, crystallized and cocrystallized with ATP. Both crystal forms belong to the trigonal space group P3{sub 2}21 and diffract X-rays to 2.0 and 2.3 Å resolution, respectively. Circadian (daily) protein clocks are found in cyanobacteria, where a complex of the KaiA, KaiB and KaiC proteins generates circadian rhythms. The 28.09 kDa KaiC homologue PH0284 protein from Pyrococcus horikoshii OT3 was cloned and expressed and the purified protein was crystallized by the oil-microbatch method at 295 K. X-ray diffraction data from the crystal were collected to 2.0 Å resolution using synchrotron radiation at 100 K. The crystal belongs to the trigonal space group P3{sub 2}21, with unit-cell parameters a = b = 96.06, c = 298.90 Å. Assuming the presence of one hexamer in the asymmetric unit gives a V{sub M} value of 2.36 Å{sup 3} Da{sup −1} and a solvent content of 47.9%. A cocrystal with ATP was prepared and a diffraction data set was collected at 2.3 Å resolution.

  15. Purification, crystallization and preliminary crystallographic analysis of archaeal 6-pyruvoyl tetrahydrobiopterin synthase homologue PH0634 from Pyrococcus horikoshii OT3.

    PubMed

    Bagautdinov, Bagautdin; Sugahara, Mitsuaki; Kunishima, Naoki

    2007-01-01

    6-Pyruvoyl tetrahydrobiopterin synthase (PTPS) catalyses the conversion of dihydroneopterin triphosphate to 6-pyruvoyl tetrahydropterin, the second of the three enzymatic steps in the synthesis of tetrahydrobiopterin from GTP. PH0634, a 13.51 kDa archaeal PTPS homologue from Pyrococcus horikoshii OT3, was overexpressed as native and selenomethionine-substituted protein and the purified protein was crystallized by the oil-microbatch method at 295 K. X-ray diffraction data were collected to 2.1 A resolution from the native crystal using synchrotron radiation at 100 K. The crystal belongs to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 35.83, b = 95.71, c = 105.65 A. Threefold noncrystallographic symmetry was identified from self-rotation calculations. Assuming the presence of a trimer in the asymmetric unit, the solvent content is 45% (V(M) = 2.24 A3 Da(-1)). The selenomethionine-substituted crystal is isomorphous to the native crystal and diffracts X-rays to 2.9 A. PMID:17183164

  16. Purification, crystallization and preliminary crystallographic analysis of RecA superfamily ATPase PH0284 from Pyrococcus horikoshii OT3.

    PubMed

    Bagautdinov, Bagautdin; Kunishima, Naoki

    2006-04-01

    Circadian (daily) protein clocks are found in cyanobacteria, where a complex of the KaiA, KaiB and KaiC proteins generates circadian rhythms. The 28.09 kDa KaiC homologue PH0284 protein from Pyrococcus horikoshii OT3 was cloned and expressed and the purified protein was crystallized by the oil-microbatch method at 295 K. X-ray diffraction data from the crystal were collected to 2.0 angstroms resolution using synchrotron radiation at 100 K. The crystal belongs to the trigonal space group P3(2)21, with unit-cell parameters a = b = 96.06, c = 298.90 angstroms. Assuming the presence of one hexamer in the asymmetric unit gives a V(M) value of 2.36 angstroms3 Da(-1) and a solvent content of 47.9%. A cocrystal with ATP was prepared and a diffraction data set was collected at 2.3 angstroms resolution. PMID:16582499

  17. Purification, crystallization and preliminary crystallographic analysis of archaeal 6-pyruvoyl tetrahydrobiopterin synthase homologue PH0634 from Pyrococcus horikoshii OT3

    SciTech Connect

    Bagautdinov, Bagautdin; Sugahara, Mitsuaki; Kunishima, Naoki

    2007-01-01

    An archaeal 6-pyruvoyl tetrahydrobiopterin synthase homologue from P. horikoshii OT3 was overexpressed as native and selenomethionine-substituted protein, purified and crystallized. The native and selenomethionine-derivative crystals are isomorphous and diffract X-rays to 2.1 and 2.9 Å resolution, respectively. 6-Pyruvoyl tetrahydrobiopterin synthase (PTPS) catalyses the conversion of dihydroneopterin triphosphate to 6-pyruvoyl tetrahydropterin, the second of the three enzymatic steps in the synthesis of tetrahydrobiopterin from GTP. PH0634, a 13.51 kDa archaeal PTPS homologue from Pyrococcus horikoshii OT3, was overexpressed as native and selenomethionine-substituted protein and the purified protein was crystallized by the oil-microbatch method at 295 K. X-ray diffraction data were collected to 2.1 Å resolution from the native crystal using synchrotron radiation at 100 K. The crystal belongs to the orthorhombic space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 35.83, b = 95.71, c = 105.65 Å. Threefold noncrystallographic symmetry was identified from self-rotation calculations. Assuming the presence of a trimer in the asymmetric unit, the solvent content is 45% (V{sub M} = 2.24 Å{sup 3} Da{sup −1}). The selenomethionine-substituted crystal is isomorphous to the native crystal and diffracts X-rays to 2.9 Å.

  18. Purification, crystallization and preliminary crystallographic analysis of archaeal 6-pyruvoyl tetrahydrobiopterin synthase homologue PH0634 from Pyrococcus horikoshii OT3

    PubMed Central

    Bagautdinov, Bagautdin; Sugahara, Mitsuaki; Kunishima, Naoki

    2007-01-01

    6-Pyruvoyl tetrahydrobiopterin synthase (PTPS) catalyses the conversion of dihydroneopterin triphosphate to 6-pyruvoyl tetrahydropterin, the second of the three enzymatic steps in the synthesis of tetrahydrobiopterin from GTP. PH0634, a 13.51 kDa archaeal PTPS homologue from Pyrococcus horikoshii OT3, was overexpressed as native and selenomethionine-substituted protein and the purified protein was crystallized by the oil-microbatch method at 295 K. X-ray diffraction data were collected to 2.1 Å resolution from the native crystal using synchrotron radiation at 100 K. The crystal belongs to the orthorhombic space group P212121, with unit-cell parameters a = 35.83, b = 95.71, c = 105.65 Å. Threefold noncrystallographic symmetry was identified from self-rotation calculations. Assuming the presence of a trimer in the asymmetric unit, the solvent content is 45% (V M = 2.24 Å3 Da−1). The selenomethionine-substituted crystal is isomorphous to the native crystal and diffracts X-rays to 2.9 Å. PMID:17183164

  19. Purification, crystallization and preliminary crystallographic analysis of the biotin-protein ligase from Pyrococcus horikoshii OT3.

    PubMed

    Bagautdinov, Bagautdin; Kuroishi, Chizu; Sugahara, Mitsuaki; Kunishima, Naoki

    2005-02-01

    Biotin-protein ligase is an enzyme that catalyzes the ATP-dependent biotinylation of a specific lysine residue in acetyl-CoA carboxylase. The biotin-protein ligase from Pyrococcus horikoshii OT3 has been cloned, overexpressed and purified. Crystallization was performed by the microbatch method or the vapour-diffusion method using PEG 2000 as a precipitant at 295 K. X-ray diffraction data have been collected to 1.6 A resolution from a native crystal and to 1.55 A resolution from a selenomethionine-derivative crystal for multiple anomalous dispersion phasing using synchrotron radiation at 100 K. The native crystal belongs to the monoclinic space group P2(1), with unit-cell parameters a = 38.601, b = 78.264, c = 70.147 A, beta = 101.48 degrees. Assuming a homodimer per asymmetric unit gives a VM value of 2.14 A3 Da(-1) and a solvent content of 42.5%. Cocrystals with biotin, ADP and biotinyl-5'-AMP were prepared and diffraction data sets were collected to 1.6, 1.6 and 1.45 A resolution, respectively. PMID:16510991

  20. Purification, crystallization and preliminary crystallographic analysis of the biotin–protein ligase from Pyrococcus horikoshii OT3

    PubMed Central

    Bagautdinov, Bagautdin; Kuroishi, Chizu; Sugahara, Mitsuaki; Kunishima, Naoki

    2005-01-01

    Biotin–protein ligase is an enzyme that catalyzes the ATP-dependent biotinylation of a specific lysine residue in acetyl-CoA carboxylase. The biotin–protein ligase from Pyrococcus horikoshii OT3 has been cloned, overexpressed and purified. Crystallization was performed by the microbatch method or the vapour-diffusion method using PEG 2000 as a precipitant at 295 K. X-ray diffraction data have been collected to 1.6 Å resolution from a native crystal and to 1.55 Å resolution from a selenomethionine-derivative crystal for multiple anomalous dispersion phasing using synchrotron radiation at 100 K. The native crystal belongs to the monoclinic space group P21, with unit-cell parameters a = 38.601, b = 78.264, c  =  70.147 Å, β = 101.48°. Assuming a homodimer per asymmetric unit gives a V M value of 2.14 Å3 Da−1 and a solvent content of 42.5%. Cocrystals with biotin, ADP and biotinyl-5′-AMP were prepared and diffraction data sets were collected to 1.6, 1.6 and 1.45 Å resolution, respectively. PMID:16510991

  1. Purification, crystallization and preliminary crystallographic analysis of RecA superfamily ATPase PH0284 from Pyrococcus horikoshii OT3

    PubMed Central

    Bagautdinov, Bagautdin; Kunishima, Naoki

    2006-01-01

    Circadian (daily) protein clocks are found in cyanobacteria, where a complex of the KaiA, KaiB and KaiC proteins generates circadian rhythms. The 28.09 kDa KaiC homologue PH0284 protein from Pyrococcus horikoshii OT3 was cloned and expressed and the purified protein was crystallized by the oil-microbatch method at 295 K. X-ray diffraction data from the crystal were collected to 2.0 Å resolution using synchrotron radiation at 100 K. The crystal belongs to the trigonal space group P3221, with unit-cell parameters a = b = 96.06, c = 298.90 Å. Assuming the presence of one hexamer in the asymmetric unit gives a V M value of 2.36 Å3 Da−1 and a solvent content of 47.9%. A cocrystal with ATP was prepared and a diffraction data set was collected at 2.3 Å resolution. PMID:16582499

  2. 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.

  3. 1989 Architectural Exhibition Winners.

    ERIC Educational Resources Information Center

    School Business Affairs, 1990

    1990-01-01

    Winners of the 1989 Architectural Exhibition sponsored annually by the ASBO International's School Facilities Research Committee include the Brevard Performing Arts Center (Melbourne, Florida), the Capital High School (Santa Fe, New Mexico), Gage Elementary School (Rochester, Minnesota), the Lakewood (Ohio) High School Natatorium, and three other…

  4. Exhibitions in Sight

    ERIC Educational Resources Information Center

    Wasserman, Burton

    1978-01-01

    During this past year a vast number of art shows have been exhibited across the United States. Their most striking features were their range and diversity. Here are some comments on Ben Shahn's paintings and photography focusing on social realism, some works by the Polish Constructivists, interested in redefining form in relation to space, the…

  5. Skylab Exhibit Ribbon Cutting

    NASA Technical Reports Server (NTRS)

    1976-01-01

    A metal strap became tangled over one of the folded solar array panels when Skylab lost its micro meteoroid shield during its launch. Cutters like the ones used to free the solar array were used to cut the ribbon opening to the public a new full-scale Skylab cluster exhibit at the Alabama Space and Rocket Center in Huntsville, Alabama. Wielding the cutters are (left to right): Alabama Senator James B. Allen; Marshall Space Flight Center director, Dr. William R. Lucas, Huntsville Mayor, Joe Davis; Madison County Commission Chairman, James Record (standing behind Mayor Davis); and chairman of the Alabama Space Science Exhibit Commission, Jack Giles. Astronauts Conrad and Kerwin used the same type of tool in Earth orbit to cut the aluminum strap which jammed the Skylab solar array.

  6. 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.

  7. 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

  8. 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.

  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. 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.

  11. A Unique Chitinase with Dual Active Sites and Triple Substrate Binding Sites from the Hyperthermophilic Archaeon Pyrococcus kodakaraensis KOD1

    PubMed Central

    Tanaka, Takeshi; Fujiwara, Shinsuke; Nishikori, Shingo; Fukui, Toshiaki; Takagi, Masahiro; Imanaka, Tadayuki

    1999-01-01

    We have found that the hyperthermophilic archaeon Pyrococcus kodakaraensis KOD1 produces an extracellular chitinase. The gene encoding the chitinase (chiA) was cloned and sequenced. The chiA gene was found to be composed of 3,645 nucleotides, encoding a protein (1,215 amino acids) with a molecular mass of 134,259 Da, which is the largest among known chitinases. Sequence analysis indicates that ChiA is divided into two distinct regions with respective active sites. The N-terminal and C-terminal regions show sequence similarity with chitinase A1 from Bacillus circulans WL-12 and chitinase from Streptomyces erythraeus (ATCC 11635), respectively. Furthermore, ChiA possesses unique chitin binding domains (CBDs) (CBD1, CBD2, and CBD3) which show sequence similarity with cellulose binding domains of various cellulases. CBD1 was classified into the group of family V type cellulose binding domains. In contrast, CBD2 and CBD3 were classified into that of the family II type. chiA was expressed in Escherichia coli cells, and the recombinant protein was purified to homogeneity. The optimal temperature and pH for chitinase activity were found to be 85°C and 5.0, respectively. Results of thin-layer chromatography analysis and activity measurements with fluorescent substrates suggest that the enzyme is an endo-type enzyme which produces a chitobiose as a major end product. Various deletion mutants were constructed, and analyses of their enzyme characteristics revealed that both the N-terminal and C-terminal halves are independently functional as chitinases and that CBDs play an important role in insoluble chitin binding and hydrolysis. Deletion mutants which contain the C-terminal half showed higher thermostability than did N-terminal-half mutants and wild-type ChiA. PMID:10583986

  12. Crystal structures of biotin protein ligase from Pyrococcus horikoshii OT3 and its complexes: structural basis of biotin activation.

    PubMed

    Bagautdinov, Bagautdin; Kuroishi, Chizu; Sugahara, Mitsuaki; Kunishima, Naoki

    2005-10-21

    Biotin protein ligase (EC 6.3.4.15) catalyses the synthesis of an activated form of biotin, biotinyl-5'-AMP, from substrates biotin and ATP followed by biotinylation of the biotin carboxyl carrier protein subunit of acetyl-CoA carboxylase. The three-dimensional structure of biotin protein ligase from Pyrococcus horikoshii OT3 has been determined by X-ray diffraction at 1.6A resolution. The structure reveals a homodimer as the functional unit. Each subunit contains two domains, a larger N-terminal catalytic domain and a smaller C-terminal domain. The structural feature of the active site has been studied by determination of the crystal structures of complexes of the enzyme with biotin, ADP and the reaction intermediate biotinyl-5'-AMP at atomic resolution. This is the first report of the liganded structures of biotin protein ligase with nucleotide and biotinyl-5'-AMP. The structures of the unliganded and the liganded forms are isomorphous except for an ordering of the active site loop upon ligand binding. Catalytic binding sites are suitably arranged to minimize the conformational changes required during the reaction, as the pockets for biotin and nucleotide are located spatially adjacent to each other in a cleft of the catalytic domain and the pocket for biotinyl-5'-AMP binding mimics the combination of those of the substrates. The exact locations of the ligands and the active site residues allow us to propose a general scheme for the first step of the reaction carried out by biotin protein ligase in which the positively charged epsilon-amino group of Lys111 facilitates the nucleophilic attack on the ATP alpha-phosphate group by the biotin carboxyl oxygen atom and stabilizes the negatively charged intermediates. PMID:16169557

  13. 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

  14. The K-loop, a general feature of the Pyrococcus C/D guide RNAs, is an RNA structural motif related to the K-turn

    PubMed Central

    Nolivos, Sophie; Carpousis, Agamemnon J.; Clouet-d'Orval, Béatrice

    2005-01-01

    The C/D guide RNAs predicted from the genomic sequences of three species of Pyrococcus delineate a family of small non-coding archaeal RNAs involved in the methylation of rRNA and tRNA. The C/D guides assemble into ribonucleoprotein (RNP) that contains the methyltransferase. The protein L7Ae, a key structural component of the RNP, binds to a Kink-turn (K-turn) formed by the C/D motif. The K-turn is a structure that consists of two RNA stems separated by a short asymmetric loop with a characteristic sharp bend (kink) between the two stems. The majority of the pyrococcal C/D guides contain a short 3 nt-spacer between the C′/D′ motifs. We show here that conserved terminal stem–loops formed by the C′/D′ motif of the Pyrococcus C/D RNAs are also L7Ae-binding sites. These stem–loops are related to the K-turn by sequence and structure, but they consist of a single stem closed by a terminal loop. We have named this structure the K-loop. We show that conserved non-canonical base pairs in the stem of the K-loop are necessary for L7Ae binding. For the C/D guides with a 3 nt-spacer we show that the sequence and length is also important. The K-loop could improve the stability of the C/D guide RNAs in Pyrococcal species, which are extreme hyperthermophiles. PMID:16293637

  15. 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

  16. Traveling Exhibitions: translating current science into effective science exhibitions

    NASA Astrophysics Data System (ADS)

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

    The Space Science Institute (SSI) of Boulder, Colorado has recently developed two museum exhibits called the Space Weather Center and MarsQuest. It is currently planning to develop two other exhibitions called Cosmic Origins and InterActive Earth. Museum exhibitions provide research scientists the opportunity to engage in a number of activities that are vital to the success of earth and space outreach programs. The Space Weather Center was developed in partnership with various research missions at NASA's Goddard Space Flight Center. The focus of the presentation will be on the Institute's MarsQuest exhibition. This project is a 5000 square-foot, 2.5M, traveling exhibition that is now touring the country. The exhibit's 3-year tour is enabling 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 host museum staff (e.g. museum educators and docents). The workshops make innovative connections between the exhibitions interactive experiences and lesson plans aligned with the National Science Education Standards. SSI is also developing an interactive web site called MarsQuest On-line. The linkage between the web site, education program and exhibit will be discussed. MarsQuest and SSI's other exhibitions 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.

  17. 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…

  18. Against the Odds Exhibition Opens

    MedlinePlus

    ... the National Institutes of Health in Bethesda, Md. Photo courtesy of Bill Branson NIH Director Dr. Elias ... addresses visitors to the opening of the exhibition. Photo courtesy of Bill Branson Brothers Niko and Theo ...

  19. Against the Odds Exhibition Opens

    MedlinePlus

    ... Bar Home Current Issue Past Issues Special Section Against the Odds Exhibition Opens Past Issues / Spring 2008 ... Research in Bethesda. Photo courtesy of Bill Branson "Against the Odds" is a remarkable story of achievement ...

  20. 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.

  1. 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.

  2. In vitro stabilization and in vivo solubilization of foreign proteins by the beta subunit of a chaperonin from the hyperthermophilic archaeon Pyrococcus sp. strain KOD1.

    PubMed Central

    Yan, Z; Fujiwara, S; Kohda, K; Takagi, M; Imanaka, T

    1997-01-01

    The gene encoding the beta subunit of a molecular chaperonin from the hyperthermophilic archaeon Pyrococcus sp. strain KOD1 (cpkB) was cloned, sequenced, and expressed in Escherichia coli. The cpkB gene is composed of 1,641 nucleotides, encoding a protein (546 amino acids) with a molecular mass of 59,140 Da. The enhancing effect of CpkB on enzyme stability was examined by using Saccharomyces cerevisiae alcohol dehydrogenase (ADH). Purified recombinant CpkB prevents thermal denaturation and enhances thermostability of ADH. CpkB requires ATP for its chaperonin function at a low CpkB concentration; however, CpkB functions without ATP when present in excess. In vivo chaperonin function for the solubilization of insoluble proteins was also studied by coexpressing CpkB and CobQ (cobryic acid synthase), indicating that CpkB is useful for solubilizing the insoluble proteins in vivo. These results suggest that the beta subunit plays a major role in chaperonin activity and is functional without the alpha subunit. PMID:9023959

  3. Comparative Physiological Studies on Hyperthermophilic Archaea Isolated from Deep-Sea Hot Vents with Emphasis on Pyrococcus Strain GB-D †

    PubMed Central

    Jannasch, Holger W.; Wirsen, Carl O.; Molyneaux, Stephen J.; Langworthy, Thomas A.

    1992-01-01

    Three new sulfur- or non-sulfur-dependent archaeal isolates, including a Pyrococcus strain, from Guaymas Basin hydrothermal vents (Gulf of California; depth, 2,010 m) were characterized and physiologically compared with four known hyperthermophiles, previously isolated from other vent sites, with an emphasis on growth and survival under the conditions particular to the natural habitat. Incubation under in situ pressure (200 atm [1 atm = 101.29 kPa]) did not increase the maximum growth temperature by more than 1°C for any of the organisms but did result in increases in growth rates of up to 15% at optimum growth temperatures. At in situ pressure, temperatures considerably higher than those limiting growth (i.e., > 105°C) were survived best by isolates with the highest maximum growth temperatures, but none of the organisms survived at temperatures of 150°C or higher for 5 min. Free oxygen was toxic to all isolates at growth range temperatures, but at ambient deep-sea temperature (3 to 4°C), the effect varied in different isolates, the non-sulfur-dependent isolate being the most oxygen tolerant. Hyperthermophiles could be isolated from refrigerated and oxygenated samples after 5 years of storage. Cu, Zn, and Pb ions were found to be toxic under nongrowth conditions (absence of organic substrate), with the non-sulfur-dependent isolate again being the most tolerant. PMID:16348799

  4. Expression, purification, crystallization and preliminary X-ray analysis of the KaiC-like protein PH0187 from the hyperthermophilic archaeon Pyrococcus horikoshii OT3

    PubMed Central

    Kang, Hee-Jin; Kubota, Keiko; Miyazono, Ken-ichi; Tanokura, Masaru

    2011-01-01

    KaiC is the central protein in the circadian rhythm in cyanobacteria. The 28 kDa KaiC-like protein PH0187 from the hyperthermophilic archaeon Pyrococcus horikoshii was expressed in Escherichia coli, purified and crystallized using the sitting-drop vapour-diffusion method at 293 K. Crystals of PH0187 were obtained using a reservoir solution consisting of 1.0 M ammonium phosphate monobasic and 0.1 M sodium citrate tribasic pH 5.3 (the final pH value of the reservoir solution was 4.8) and diffracted X-rays to 2.75 Å resolution. The crystal of PH0187 belonged to space group P6322, with unit-cell parameters a = b = 239.1, c = 106.5 Å. The crystal contained four PH0187 molecules in the asymmetric unit. PMID:21206047

  5. Crystallization and preliminary X-ray crystallographic studies of the biotin carboxyl carrier protein and biotin protein ligase complex from Pyrococcus horikoshii OT3.

    PubMed

    Bagautdinov, Bagautdin; Matsuura, Yoshinori; Bagautdinova, Svetlana; Kunishima, Naoki

    2007-04-01

    Biotin protein ligase (BPL) catalyses the biotinylation of the biotin carboxyl carrier protein (BCCP) subunit of acetyl-CoA carboxylase. To elucidate the exact details of the protein-protein interactions in the biotinylation function, the C-terminal half fragment of BCCP (BCCPDeltaN76), the R48A mutant of BPL (BPL*) and the R48A K111A double mutant of BPL (BPL**), all of which are from Pyrococcus horikoshii OT3, have been expressed, purified and successfully cocrystallized. Cocrystals of the BPL*-BCCPDeltaN76 and BPL**-BCCPDeltaN76 complexes as well as crystals of BPL*, BPL** and BCCPDeltaN76 were obtained by the oil-microbatch method using PEG 20 000 as a precipitant at 295 K. Complete X-ray diffraction data sets for BPL*-BCCPDeltaN76 and BPL**-BCCPDeltaN76 crystals were collected at 100 K to 2.7 and 2.0 A resolution, respectively, using synchrotron radiation. They belong to the monoclinic space group P2(1), with similar unit-cell parameters a = 69.85, b = 63.12, c = 75.64 A, beta = 95.9 degrees . Assuming two subunits of the complex per asymmetric unit gives a V(M) value of 2.45 A(3) Da(-1) and a solvent content of 50%. PMID:17401210

  6. Crystallization and preliminary X-ray crystallographic studies of the biotin carboxyl carrier protein and biotin protein ligase complex from Pyrococcus horikoshii OT3

    PubMed Central

    Bagautdinov, Bagautdin; Matsuura, Yoshinori; Bagautdinova, Svetlana; Kunishima, Naoki

    2007-01-01

    Biotin protein ligase (BPL) catalyses the biotinylation of the biotin carboxyl carrier protein (BCCP) subunit of acetyl-CoA carboxylase. To elucidate the exact details of the protein–protein interactions in the biotinylation function, the C-terminal half fragment of BCCP (BCCPΔN76), the R48A mutant of BPL (BPL*) and the R48A K111A double mutant of BPL (BPL**), all of which are from Pyrococcus horikoshii OT3, have been expressed, purified and successfully cocrystallized. Cocrystals of the BPL*–BCCPΔN76 and BPL**–BCCPΔN76 complexes as well as crystals of BPL*, BPL** and BCCPΔN76 were obtained by the oil-microbatch method using PEG 20 000 as a precipitant at 295 K. Complete X-ray diffraction data sets for BPL*–BCCPΔN76 and BPL**–BCCPΔN76 crystals were collected at 100 K to 2.7 and 2.0 Å resolution, respectively, using synchrotron radiation. They belong to the monoclinic space group P21, with similar unit-cell parameters a = 69.85, b = 63.12, c = 75.64 Å, β = 95.9°. Assuming two subunits of the complex per asymmetric unit gives a V M value of 2.45 Å3 Da−1 and a solvent content of 50%. PMID:17401210

  7. 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)

  8. The structures of the CutA1 proteins from Thermus thermophilus and Pyrococcus horikoshii: characterization of metal-binding sites and metal-induced assembly

    PubMed Central

    Bagautdinov, Bagautdin

    2014-01-01

    CutA1 (copper tolerance A1) is a widespread cytoplasmic protein found in archaea, bacteria, plants and animals, including humans. In Escherichia coli it is implicated in divalent metal tolerance, while the mammalian CutA1 homologue has been proposed to mediate brain enzyme acetylcholinesterase activity and copper homeostasis. The X-ray structures of CutA1 from the thermophilic bacterium Thermus thermophilus (TtCutA1) with and without bound Na+ at 1.7 and 1.9 Å resolution, respectively, and from the hyperthermophilic archaeon Pyrococcus horikoshii (PhCutA1) in complex with Na+ at 1.8 Å resolution have been determined. Both are short and rigid proteins of about 12 kDa that form intertwined compact trimers in the crystal and solution. The main difference in the structures is a wide-type β-bulge on top of the TtCutA1 trimer. It affords a mechanism for lodging a single-residue insertion in the middle of β2 while preserving the interprotomer main-chain hydrogen-bonding network. The liganded forms of the proteins provide new structural information about the metal-binding sites and CutA1 assembly. The Na+–TtCutA1 structure unveils a dodecameric assembly with metal ions in the trimer–trimer interfaces and the lateral clefts of the trimer. For Na+–PhCutA1, the metal ion associated with six waters in an octahedral geometry. The structures suggest that CutA1 may contribute to regulating intracellular metal homeostasis through various binding modes. PMID:24699729

  9. The structures of the CutA1 proteins from Thermus thermophilus and Pyrococcus horikoshii: characterization of metal-binding sites and metal-induced assembly.

    PubMed

    Bagautdinov, Bagautdin

    2014-04-01

    CutA1 (copper tolerance A1) is a widespread cytoplasmic protein found in archaea, bacteria, plants and animals, including humans. In Escherichia coli it is implicated in divalent metal tolerance, while the mammalian CutA1 homologue has been proposed to mediate brain enzyme acetylcholinesterase activity and copper homeostasis. The X-ray structures of CutA1 from the thermophilic bacterium Thermus thermophilus (TtCutA1) with and without bound Na(+) at 1.7 and 1.9 Å resolution, respectively, and from the hyperthermophilic archaeon Pyrococcus horikoshii (PhCutA1) in complex with Na(+) at 1.8 Å resolution have been determined. Both are short and rigid proteins of about 12 kDa that form intertwined compact trimers in the crystal and solution. The main difference in the structures is a wide-type β-bulge on top of the TtCutA1 trimer. It affords a mechanism for lodging a single-residue insertion in the middle of β2 while preserving the interprotomer main-chain hydrogen-bonding network. The liganded forms of the proteins provide new structural information about the metal-binding sites and CutA1 assembly. The Na(+)-TtCutA1 structure unveils a dodecameric assembly with metal ions in the trimer-trimer interfaces and the lateral clefts of the trimer. For Na(+)-PhCutA1, the metal ion associated with six waters in an octahedral geometry. The structures suggest that CutA1 may contribute to regulating intracellular metal homeostasis through various binding modes. PMID:24699729

  10. Longwall - USA: International exhibition & conference

    SciTech Connect

    1996-12-31

    The Longwall-USA International Exhibition and Conference was held June 4-6, 1996 in Pittsburgh, PA. Seventeen papers are included in the proceedings that covered such topics as health and safety, development of gate roads, telemetry monitoring systems, fires, longwall miners, roof support technologies, dust control, moving car bunker systems, reducing longwall noise, vibration of longwall equipment, and the USBM`s strategic structures testing laboratory. A separate abstract with indexing was prepared for each paper for inclusion in the Energy Science and Technology Database.

  11. 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.

  12. 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

  13. Cystamine preparations exhibit anticoagulant activity.

    PubMed

    Aleman, Maria M; Holle, Lori A; Stember, Katherine G; Devette, Christa I; Monroe, Dougald M; Wolberg, Alisa S

    2015-01-01

    Transglutaminases are a superfamily of isoenzymes found in cells and plasma. These enzymes catalyze the formation of ε-N-(γ-glutamyl)-lysyl crosslinks between proteins. Cystamine blocks transglutaminase activity and is used in vitro in human samples and in vivo in mice and rats in studies of coagulation, immune dysfunction, and inflammatory disease. These studies have suggested cystamine blocks fibrin crosslinking and has anti-inflammatory effects, implicating transglutaminase activity in the pathogenesis of several diseases. We measured the effects of cystamine on fibrin crosslinking, tissue factor-triggered plasma clot formation and thrombin generation, and coagulation factor enzymatic activity. At concentrations that blocked fibrin crosslinking, cystamine also inhibited plasma clot formation and reduced thrombin generation. Cystamine inhibited the amidolytic activity of coagulation factor XI and thrombin towards chromogenic substrates. These findings demonstrate that cystamine exhibits anticoagulant activity during coagulation. Given the close relationship between coagulation and inflammation, these findings suggest prior studies that used cystamine to implicate transglutaminase activity in disease pathogenesis warrant re-examination. PMID:25915545

  14. Cystamine Preparations Exhibit Anticoagulant Activity

    PubMed Central

    Aleman, Maria M.; Holle, Lori A.; Stember, Katherine G.; Devette, Christa I.; Monroe, Dougald M.; Wolberg, Alisa S.

    2015-01-01

    Transglutaminases are a superfamily of isoenzymes found in cells and plasma. These enzymes catalyze the formation of ε-N-(γ-glutamyl)-lysyl crosslinks between proteins. Cystamine blocks transglutaminase activity and is used in vitro in human samples and in vivo in mice and rats in studies of coagulation, immune dysfunction, and inflammatory disease. These studies have suggested cystamine blocks fibrin crosslinking and has anti-inflammatory effects, implicating transglutaminase activity in the pathogenesis of several diseases. We measured the effects of cystamine on fibrin crosslinking, tissue factor-triggered plasma clot formation and thrombin generation, and coagulation factor enzymatic activity. At concentrations that blocked fibrin crosslinking, cystamine also inhibited plasma clot formation and reduced thrombin generation. Cystamine inhibited the amidolytic activity of coagulation factor XI and thrombin towards chromogenic substrates. These findings demonstrate that cystamine exhibits anticoagulant activity during coagulation. Given the close relationship between coagulation and inflammation, these findings suggest prior studies that used cystamine to implicate transglutaminase activity in disease pathogenesis warrant re-examination. PMID:25915545

  15. Crystallization and preliminary X-ray crystallographic studies of the biotin carboxyl carrier protein and biotin protein ligase complex from Pyrococcus horikoshii OT3

    SciTech Connect

    Bagautdinov, Bagautdin; Matsuura, Yoshinori; Bagautdinova, Svetlana; Kunishima, Naoki

    2007-04-01

    A truncated form of biotin carboxyl carrier protein containing the C-terminal half fragment (BCCPΔN76) and the biotin protein ligase (BPL) with the mutation R48A (BPL*) or the double mutation R48A K111A (BPL**) were successfully cocrystallized in the presence of ATP and biotin. The BPL*–BCCPΔN76 and BPL**–BCCPΔN76 crystals belong to space group P2{sub 1} and diffract X-rays to 2.7 and 2.0 Å resolution, respectively. Biotin protein ligase (BPL) catalyses the biotinylation of the biotin carboxyl carrier protein (BCCP) subunit of acetyl-CoA carboxylase. To elucidate the exact details of the protein–protein interactions in the biotinylation function, the C-terminal half fragment of BCCP (BCCPΔN76), the R48A mutant of BPL (BPL*) and the R48A K111A double mutant of BPL (BPL**), all of which are from Pyrococcus horikoshii OT3, have been expressed, purified and successfully cocrystallized. Cocrystals of the BPL*–BCCPΔN76 and BPL**–BCCPΔN76 complexes as well as crystals of BPL*, BPL** and BCCPΔN76 were obtained by the oil-microbatch method using PEG 20 000 as a precipitant at 295 K. Complete X-ray diffraction data sets for BPL*–BCCPΔN76 and BPL**–BCCPΔN76 crystals were collected at 100 K to 2.7 and 2.0 Å resolution, respectively, using synchrotron radiation. They belong to the monoclinic space group P2{sub 1}, with similar unit-cell parameters a = 69.85, b = 63.12, c = 75.64 Å, β = 95.9°. Assuming two subunits of the complex per asymmetric unit gives a V{sub M} value of 2.45 Å{sup 3} Da{sup −1} and a solvent content of 50%.

  16. 29 CFR 2200.70 - Exhibits.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 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 petition... proceeding. (f) Request for custody of physical exhibit. Any person may on motion to the Executive...

  17. 29 CFR 2200.70 - Exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 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 petition... proceeding. (f) Request for custody of physical exhibit. Any person may on motion to the Executive...

  18. 29 CFR 2200.70 - Exhibits.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 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 petition... proceeding. (f) Request for custody of physical exhibit. Any person may on motion to the Executive...

  19. 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…

  20. An Astrobiology Microbes Exhibit and Education Module

    NASA Astrophysics Data System (ADS)

    Lindstrom, M. M.; Allen, J. S.; Mortillaro, C.; Ducceschi, C.; Rawlings, P.; Stocco, K.; Tobola, K.; Olendzenski, L.; Angermiller, L.

    2001-03-01

    A JSC-SCH team has produced an astrobiology exhibit and education module to augment the 'Microbes!' traveling exhibit. The astrobiology section focuses on life in extreme environments and considers the possibility of extraterrestrrial life.

  1. 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)

  2. 49 CFR 1114.7 - Exhibits.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... practical the sheets of each exhibit and the lines of each sheet should be numbered. If the exhibit consists of five or more sheets, the first sheet or title-page should be confined to a brief statement of what the exhibit purports to show with reference by sheet and line to illustrative or typical...

  3. 49 CFR 1114.7 - Exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... practical the sheets of each exhibit and the lines of each sheet should be numbered. If the exhibit consists of five or more sheets, the first sheet or title-page should be confined to a brief statement of what the exhibit purports to show with reference by sheet and line to illustrative or typical...

  4. 49 CFR 1114.7 - Exhibits.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... practical the sheets of each exhibit and the lines of each sheet should be numbered. If the exhibit consists of five or more sheets, the first sheet or title-page should be confined to a brief statement of what the exhibit purports to show with reference by sheet and line to illustrative or typical...

  5. 49 CFR 1114.7 - Exhibits.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... practical the sheets of each exhibit and the lines of each sheet should be numbered. If the exhibit consists of five or more sheets, the first sheet or title-page should be confined to a brief statement of what the exhibit purports to show with reference by sheet and line to illustrative or typical...

  6. 49 CFR 1114.7 - Exhibits.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... practical the sheets of each exhibit and the lines of each sheet should be numbered. If the exhibit consists of five or more sheets, the first sheet or title-page should be confined to a brief statement of what the exhibit purports to show with reference by sheet and line to illustrative or typical...

  7. 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.

  8. A euryarchaeal histone modulates strand displacement synthesis by replicative DNA polymerases.

    PubMed

    Sun, Fei; Huang, Li

    2016-07-01

    Euryarchaeota and Crenarchaeota, the two main lineages of the domain Archaea, encode different chromatin proteins and differ in the use of replicative DNA polymerases. Crenarchaea possess a single family B DNA polymerase (PolB), which is capable of strand displacement modulated by the chromatin proteins Cren7 and Sul7d. Euryarchaea have two distinct replicative DNA polymerases, PolB and PolD, a family D DNA polymerase. Here we characterized the strand displacement activities of PolB and PolD from the hyperthermophilic euryarchaeon Pyrococcus furiosus and investigated the influence of HPfA1, a homolog of eukaryotic histones from P. furiosus, on these activities. We showed that both PolB and PolD were efficient in strand displacement. HPfA1 inhibited DNA strand displacement by both DNA polymerases but exhibited little effect on the displacement of a RNA strand annealed to single-stranded template DNA. This is consistent with the finding that HPfA1 bound more tightly to double-stranded DNA than to a RNA:DNA hybrid. Our results suggest that, although crenarchaea and euryarchaea differ in chromosomal packaging, they share similar mechanisms in modulating strand displacement by DNA polymerases during lagging strand DNA synthesis. PMID:27333783

  9. 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.

  10. 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…

  11. 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".

  12. 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…

  13. An Attention Model for Museum Exhibits.

    ERIC Educational Resources Information Center

    Lightner, John W.

    A qualitative study determined which factors in the museum exhibit environment or within the museum visitor may influence the visitor to attend an exhibit. Observations and interviews were conducted of 14 groups that visited a Chesapeake & Ohio steam locomotive at the Henry Ford Museum in Dearborn, Michigan. An inductive or grounded theory…

  14. 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…

  15. 18 CFR 156.5 - Exhibits.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... (respondent) necessary for the rendition of service requested by the applicant. (6) Exhibit G—Flow diagram showing daily design capacity and reflecting operation with proposed transmission facilities. A flow...—Flow diagram reflecting maximum capabilities. If Exhibit G does not reflect the maximum deliveries...

  16. 18 CFR 156.5 - Exhibits.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... (respondent) necessary for the rendition of service requested by the applicant. (6) Exhibit G—Flow diagram showing daily design capacity and reflecting operation with proposed transmission facilities. A flow...—Flow diagram reflecting maximum capabilities. If Exhibit G does not reflect the maximum deliveries...

  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

    ... 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 HOUSING... Program Pt. 1980, Subpt. E, Exh. G Exhibit G to Subpart E of Part 1980 Note: The Exhibit is not...

  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

    ... 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 HOUSING... Program Pt. 1980, Subpt. E, Exh. G Exhibit G to Subpart E of Part 1980 Note: The Exhibit is not...

  19. 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

    ... 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 HOUSING... Program Pt. 1980, Subpt. E, Exh. G Exhibit G to Subpart E of Part 1980 Note: The Exhibit is not...

  20. 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

    ... 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 HOUSING... Program Pt. 1980, Subpt. E, Exh. G Exhibit G to Subpart E of Part 1980 Note: The Exhibit is not...

  1. 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

    ... 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 HOUSING... Program Pt. 1980, Subpt. E, Exh. G Exhibit G to Subpart E of Part 1980 Note: The Exhibit is not...

  2. Museum Exhibitions: Optimizing Development Using Evaluation

    NASA Astrophysics Data System (ADS)

    Dusenbery, P. B.

    2002-12-01

    The Space Science Institute (SSI) of Boulder, Colorado, has recently developed two museum exhibits called the Space Weather Center and MarsQuest. It is currently planning to develop a third exhibit called InterActive Earth. The Space Weather Center was developed in partnership with various research missions at NASA's Goddard Space Flight Center. The development of these exhibitions included a comprehensive evaluation plan. I will report on the important role evaluation plays in exhibit design and development using MarsQuest and InterActive Earth as models. The centerpiece of SSI's Mars Education Program is the 5,000-square-foot traveling exhibition, MarsQuest: Exploring the Red Planet, which was developed with support from the National Science Foundation (NSF), NASA, and several corporate donors. The MarsQuest exhibit is nearing the end of a highly successful, fully-booked three-year tour. The Institute plans to send an enhanced and updated MarsQuest on a second three-year tour and is also developing Destination: Mars, a mini-version of MarsQuest designed for smaller venues. They are designed to inspire and empower participants to extend the excitement and science content of the exhibitions into classrooms and museum-based education programs in an ongoing fashion. The centerpiece of the InterActive Earth project is a traveling exhibit that will cover about 4,000 square feet. The major goal of the proposed exhibit is to introduce students and the public to the complexity of the interconnections in the Earth system, and thereby, to inspire them to better understand planet Earth. Evaluation must be an integral part of the exhibition development process. For MarsQuest, a 3-phase evaluation (front end, formative and summative) was conducted by Randi Korn and Associates in close association with the development team. Sampling procedures for all three evaluation phases ensured the participation of all audiences, including family groups, students, and adults. Each phase of

  3. 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)

  4. 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...

  5. 32 CFR 705.24 - Exhibits.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... tractor-trailer transportation should be forwarded prior to November 15th previous to the year desired. A... time for which an exhibit is authorized will be determined by the nature of the event and the type...

  6. 32 CFR 705.24 - Exhibits.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... tractor-trailer transportation should be forwarded prior to November 15th previous to the year desired. A... time for which an exhibit is authorized will be determined by the nature of the event and the type...

  7. 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)

  8. 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)

  9. 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…

  10. 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.

  11. 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."

  12. 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;

  13. 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

  14. Expression, high-pressure refolding, purification, crystallization and preliminary X-ray analysis of a novel single-strand-specific 3′–5′ exonuclease PhoExo I from Pyrococcus horikoshii OT3

    PubMed Central

    Miyazono, Ken-ichi; Tsutsumi, Kanae; Ishino, Yoshizumi; Tanokura, Masaru

    2014-01-01

    PhoExo I is a single-strand-specific 3′–5′ exonuclease from Pyrococcus horikoshii OT3 and is thought to be involved in a Thermococcales-specific DNA-repair pathway. The recombinant PhoExo I protein was produced as inclusion bodies in Escherichia coli cells. Solubilization of the inclusion bodies was performed by the high-pressure refolding method and highly purified protein was subjected to crystallization by the sitting-drop vapour-diffusion method at 20°C. A crystal of PhoExo I was obtained in a reservoir solution consisting of 0.1 M Tris–HCl pH 8.9, 27% PEG 6000 and diffracted X-rays to 1.52 Å resolution. The crystal of PhoExo I belonged to space group H32, with unit-cell parameters a = b = 112.07, c = 202.28 Å. The crystal contained two PhoExo I molecules in the asymmetric unit. PMID:25084386

  15. Expression, high-pressure refolding, purification, crystallization and preliminary X-ray analysis of a novel single-strand-specific 3'-5' exonuclease PhoExo I from Pyrococcus horikoshii OT3.

    PubMed

    Miyazono, Ken-ichi; Tsutsumi, Kanae; Ishino, Yoshizumi; Tanokura, Masaru

    2014-08-01

    PhoExo I is a single-strand-specific 3'-5' exonuclease from Pyrococcus horikoshii OT3 and is thought to be involved in a Thermococcales-specific DNA-repair pathway. The recombinant PhoExo I protein was produced as inclusion bodies in Escherichia coli cells. Solubilization of the inclusion bodies was performed by the high-pressure refolding method and highly purified protein was subjected to crystallization by the sitting-drop vapour-diffusion method at 20°C. A crystal of PhoExo I was obtained in a reservoir solution consisting of 0.1 M Tris-HCl pH 8.9, 27% PEG 6000 and diffracted X-rays to 1.52 Å resolution. The crystal of PhoExo I belonged to space group H32, with unit-cell parameters a = b = 112.07, c = 202.28 Å. The crystal contained two PhoExo I molecules in the asymmetric unit. PMID:25084386

  16. 18 CFR 156.5 - Exhibits.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... engineering design data in explanation and support of the diagrams and the proposed project, setting forth: (i... 18 Conservation of Power and Water Resources 1 2012-04-01 2012-04-01 false Exhibits. 156.5 Section 156.5 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION, DEPARTMENT...

  17. 10 CFR 205.303 - Required exhibits.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... pertinent Federal and State laws. (c) Exhibit C. A general map showing the applicant's overall electric system and a detailed map highlighting the location of the facilities or the proposed facilities to be used for the generation and transmission of the electric energy to be exported. The detailed map...

  18. 10 CFR 205.303 - Required exhibits.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... pertinent Federal and State laws. (c) Exhibit C. A general map showing the applicant's overall electric system and a detailed map highlighting the location of the facilities or the proposed facilities to be used for the generation and transmission of the electric energy to be exported. The detailed map...

  19. 18 CFR 34.4 - Required exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... stockholders has been obtained. (c) Exhibit C. The Balance Sheet and attached notes for the most recent 12... computation of interest coverage Actual for the year ended mm-dd-yy OMB control No. 1902-0043, pro forma for the year ended mm-dd-yy Net income Add: Interest on Long-Term Debt, Interest on Short-Term Debt,...

  20. 18 CFR 156.5 - Exhibits.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 18 Conservation of Power and Water Resources 1 2014-04-01 2014-04-01 false Exhibits. 156.5 Section 156.5 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION, DEPARTMENT OF... outlet connections at each compressor station. (iv) Pressures and volumes of gas at each intake...

  1. 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…

  2. Do Online Students Exhibit Different Learning Styles

    ERIC Educational Resources Information Center

    Hausler, Joel; Sanders, John W.; Young, Barbara

    2007-01-01

    We examined the relationship between learning styles and student type. This research seeks to examine if online students exhibit different learning styles from onsite students; and, if so, what accommodations relating to learning style differences may be made for online students? Students (N = 80) were asked to complete an online survey in order…

  3. 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

  4. 49 CFR 250.2 - Required exhibits.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    .... (8) As Exhibit 8, a copy of applicant's most recent year-end general balance sheet certified by... general balance sheet as of a date no less recent than the end of the third month preceding the date of the filing of the application. The unaudited balance sheet shall be presented in account form...

  5. 49 CFR 250.2 - Required exhibits.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    .... (8) As Exhibit 8, a copy of applicant's most recent year-end general balance sheet certified by... general balance sheet as of a date no less recent than the end of the third month preceding the date of the filing of the application. The unaudited balance sheet shall be presented in account form...

  6. 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 INTERSTATE ELECTRIC TRANSMISSION FACILITIES §...

  7. 32 CFR 705.24 - Exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... favorably only when not in conflict with recruiting requirements. (i) Requests for exhibits must be... among the Armed Forces, or with other agencies of the Federal Government. (i) All Navy activities will... concerned, via the chain of command. (3) The official OASD(PA) Request Form for Armed Forces...

  8. 18 CFR 50.7 - Applications: exhibits.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 18 Conservation of Power and Water Resources 1 2013-04-01 2013-04-01 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...

  9. 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, DEPARTMENT OF ENERGY REGULATIONS UNDER THE FEDERAL POWER ACT APPLICATIONS FOR PERMITS TO SITE...

  10. 18 CFR 50.7 - Applications: exhibits.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 18 Conservation of Power and Water Resources 1 2014-04-01 2014-04-01 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...

  11. 18 CFR 153.8 - Required exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... facilities in the United States and Canada or Mexico; (5) Exhibit E. If the proposal is to import or export... Seismic Risk Map of the United States, or where there is a risk of surface faulting or ground liquefaction... the Seismic Review of LNG Facilities,” NBSIR 84-2833. This document may be obtained from the...

  12. 18 CFR 156.5 - Exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... such enterprises or operations, a detailed explanation of each such relationship, including the... relationship. (5) Exhibit F—Location of facilities. A geographical map of suitable scale and detail showing all...—Construction, operation, and management. A concise statement setting forth arrangements for...

  13. 18 CFR 157.14 - Exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... explanation of each such relationship, including the percentage of voting strength represented by such... detailed explanation of each such relationship. (5) Exhibit E—Other pending applications and filings. A... abandoned. This map, or an additional map, shall clearly show the relationship of the new facilities to...

  14. 10 CFR 205.303 - Required exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...; Applications; Administrative Procedures and Sanctions Application for Authorization to Transmit Electric Energy... used for the generation and transmission of the electric energy to be exported. The detailed map shall... or fixing of rates for the purchase, sale or transmission of electric energy. (f) Exhibit F....

  15. 24 CFR 180.645 - Exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 24 Housing and Urban Development 1 2010-04-01 2010-04-01 false Exhibits. 180.645 Section 180.645... OPPORTUNITY CONSOLIDATED HUD HEARING PROCEDURES FOR CIVIL RIGHTS MATTERS Procedures at Hearing § 180.645... evidence could not reasonably be anticipated at that time. (c) Authenticity. The authenticity of...

  16. 18 CFR 157.14 - Exhibits.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... Act (42 U.S.C. 7101-7352); E.O. 12009, 3 CFR 142) Editorial Note: For Federal Register citations affecting § 157.14, see the List of CFR Sections Affected, which appears in the Finding Aids section of the...) Exhibit G—Flow diagrams showing daily design capacity and reflecting operation with and without...

  17. 18 CFR 157.14 - Exhibits.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... Act (42 U.S.C. 7101-7352); E.O. 12009, 3 CFR 142) Editorial Note: For Federal Register citations affecting § 157.14, see the List of CFR Sections Affected, which appears in the Finding Aids section of the...) Exhibit G—Flow diagrams showing daily design capacity and reflecting operation with and without...

  18. 18 CFR 157.14 - Exhibits.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... Act (42 U.S.C. 7101-7352); E.O. 12009, 3 CFR 142) Editorial Note: For Federal Register citations affecting § 157.14, see the List of CFR Sections Affected, which appears in the Finding Aids section of the...) Exhibit G—Flow diagrams showing daily design capacity and reflecting operation with and without...

  19. 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

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. A metafluid exhibiting strong optical magnetism.

    PubMed

    Sheikholeslami, Sassan N; Alaeian, Hadiseh; Koh, Ai Leen; Dionne, Jennifer A

    2013-09-11

    Advances in the field of metamaterials have enabled unprecedented control of light-matter interactions. Metamaterial constituents support high-frequency electric and magnetic dipoles, which can be used as building blocks for new materials capable of negative refraction, electromagnetic cloaking, strong visible-frequency circular dichroism, and enhancing magnetic or chiral transitions in ions and molecules. While all metamaterials to date have existed in the solid-state, considerable interest has emerged in designing a colloidal metamaterial or "metafluid". Such metafluids would combine the advantages of solution-based processing with facile integration into conventional optical components. Here we demonstrate the colloidal synthesis of an isotropic metafluid that exhibits a strong magnetic response at visible frequencies. Protein-antibody interactions are used to direct the solution-phase self-assembly of discrete metamolecules comprised of silver nanoparticles tightly packed around a single dielectric core. The electric and magnetic response of individual metamolecules and the bulk metamaterial solution are directly probed with optical scattering and spectroscopy. Effective medium calculations indicate that the bulk metamaterial exhibits a negative effective permeability and a negative refractive index at modest fill factors. This metafluid can be synthesized in large-quantity and high-quality and may accelerate development of advanced nanophotonic and metamaterial devices. PMID:23919764

  7. 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.

  8. Shape-Memory PVDF Exhibiting Switchable Piezoelectricity.

    PubMed

    Hoeher, Robin; Raidt, Thomas; Novak, Nikola; Katzenberg, Frank; Tiller, Joerg C

    2015-12-01

    In this study, a material is designed which combines the properties of shape-memory and electroactive polymers. This is achieved by covalent cross-linking of polyvinylidene fluoride. The resulting polymer network exhibits excellent shape-memory properties with a storable strain of 200%, and fixity as well as recovery values of 100%. Programming upon rolling induces the transformation from the nonelectroactive α-phase to the piezoelectric β-phase. The highest β-phase content is found to be 83% for a programming strain of 200% affording a d33 value of -30 pm V(-1). This is in good accordance with literature known values for piezoelectric properties. Thermal triggering this material does not only result in a shape change but also renders the material nonelectroactive. PMID:26332996

  9. Supercomputing meets seismology in earthquake exhibit

    ScienceCinema

    Blackwell, Matt; Rodger, Arthur; Kennedy, Tom

    2014-07-22

    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.

  10. Nanoporous frameworks exhibiting multiple stimuli responsiveness

    NASA Astrophysics Data System (ADS)

    Kundu, Pintu K.; Olsen, Gregory L.; Kiss, Vladimir; Klajn, Rafal

    2014-04-01

    Nanoporous frameworks are polymeric materials built from rigid molecules, which give rise to their nanoporous structures with applications in gas sorption and storage, catalysis and others. Conceptually new applications could emerge, should these beneficial properties be manipulated by external stimuli in a reversible manner. One approach to render nanoporous frameworks responsive to external signals would be to immobilize molecular switches within their nanopores. Although the majority of molecular switches require conformational freedom to isomerize, and switching in the solid state is prohibited, the nanopores may provide enough room for the switches to efficiently isomerize. Here we describe two families of nanoporous materials incorporating the spiropyran molecular switch. These materials exhibit a variety of interesting properties, including reversible photochromism and acidochromism under solvent-free conditions, light-controlled capture and release of metal ions, as well reversible chromism induced by solvation/desolvation.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. Extract from Ceratonia siliqua Exhibits Depigmentation Properties.

    PubMed

    Lall, Namrita; Kishore, Navneet; Momtaz, Saeideh; Hussein, Ahmed; Naidoo, Sanushka; Nqephe, Mabatho; Crampton, Bridget

    2015-11-01

    Skin hyper-pigmentation is a condition initiated by the overproduction of melanin existing in the melanocytes. Melanin pigment is responsible for the colour of skin in humans. It is formed through a series of oxidative reactions involving the amino acid tyrosine in the presence of the key enzyme tyrosinase. In continuation with our efforts to identify tyrosinase inhibitors from plants sources, the methanol extract from leaf, bark and fruit of Ceratonia siliqua were screened for tyrosinase inhibition and diphenolase activity. The bark extract exhibited significant inhibition on mushroom tyrosinase using L-tyrosine as a substrate and showed diphenolase activity. The extract further significantly lowered tyrosinase mRNA levels in B16-F10 mouse melanocytes. Bioassay-guided fractionation led to the isolation of six compounds. Compounds (-)-epicatechin-3-O-gallate, 1,2,3,6-tetra-O-galloyl-ß-D-glucose and gallocatechin-3-O-gallate showed tyrosinase inhibitions with the IC50 values of 27.52, 83.30 and 28.30 µg/mL, respectively. These compounds also exhibited L-DOPA activities with IC50 values of >200, 150 and 200 µg/mL, respectively. A clinical study was conducted using 20 volunteers in a patch testing trial for irritancy potential and skin depigmentation. The clinical results showed the sample to be non-irritant with irritancy potential of -34.21 and depigmentation trial showed an improvement in the even skin tone of UV induced pigmentation at 3% after 28 days of application. PMID:26201055

  16. 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

  17. 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

  18. Virtual Exhibition and Fruition of Archaeological Finds

    NASA Astrophysics Data System (ADS)

    Manferdini, A. M.; Garagnani, S.

    2011-09-01

    During the last two decades, since digital technologies have become more sophisticated in acquiring real data and building faithful copies of them, their improvements have suggested interesting applications in the field of valorisation of Historical, Cultural and Artistic Heritage, with significant consequences in the share and widespread of knowledge. But although several technologies and methodologies for 3d digitization have recently been developed and improved, the lack of a standard procedure and the costs connected to their use still doesn't encourage the systematic digital acquisition of wide collections and heritage. The aim of this paper is to show the state of the art of a project whose aim is to provide a methodology and a procedure to create digital reproductions of artefacts for Institutions called to preserve, manage and enhance the fruition of archaeological finds inside museums or through digital exhibitions. Our project's aim is to find the most suitable procedure to digitally acquire archaeo logical artefacts that usually have small dimensions and have very complex and detailed surfaces. Within our methodology, particular attention has been paid to the use of widely shared and open-source visualization systems that enhance the involvement of the user by emphasizing three-dimensional characteristics of artefacts through virtual reality.

  19. 14 CFR Appendix to Subpart A of... - Listing of Exhibits

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... Exhibit A—Budget Summary Exhibit B—Standard Grant and Cooperative Agreement Cover Page Exhibit C... Cooperative Agreements between NASA and the Commercial Space Centers Exhibit F—NASA 1674 Letter of Delegation... Reports Note: Exhibits are available at NASA Headquarters, Code HC, Washington, D.C. 20546....

  20. 14 CFR Appendix to Subpart A of... - Listing of Exhibits

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...—Budget Summary Exhibit B—Standard Grant and Cooperative Agreement Cover Page Exhibit C—Provisions Exhibit... Cooperative Agreements between NASA and the Commercial Space Centers Exhibit F—NASA 1674 Letter of Delegation... Reports Note: Exhibits are available at NASA Headquarters, Code HC, Washington, D.C. 20546....

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

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Exhibit A—Budget Summary Exhibit B—Standard Grant and Cooperative Agreement Cover Page Exhibit C... Cooperative Agreements between NASA and the Commercial Space Centers Exhibit F—NASA 1674 Letter of Delegation... Reports Note: Exhibits are available at NASA Headquarters, Code HC, Washington, D.C. 20546....

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

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... Exhibit A—Budget Summary Exhibit B—Standard Grant and Cooperative Agreement Cover Page Exhibit C... Cooperative Agreements between NASA and the Commercial Space Centers Exhibit F—NASA 1674 Letter of Delegation... Reports Note: Exhibits are available at NASA Headquarters, Code HC, Washington, D.C. 20546....

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

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Exhibit A—Budget Summary Exhibit B—Standard Grant and Cooperative Agreement Cover Page Exhibit C... Cooperative Agreements between NASA and the Commercial Space Centers Exhibit F—NASA 1674 Letter of Delegation... Reports Note: Exhibits are available at NASA Headquarters, Code HC, Washington, D.C. 20546....

  4. 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

  5. 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.

  6. 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. PMID:23324742

  7. 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.

  8. 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. PMID:24657593

  9. 37 CFR 1.95 - Copies of exhibits.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ..., 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...

  10. 37 CFR 1.95 - Copies of exhibits.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ..., 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...

  11. 37 CFR 1.95 - Copies of exhibits.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ..., 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...

  12. 37 CFR 1.95 - Copies of exhibits.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ..., 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...

  13. 37 CFR 1.95 - Copies of exhibits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ..., 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...

  14. 29 CFR 2204.202 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 29 Labor 9 2011-07-01 2011-07-01 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...

  15. 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...

  16. 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…

  17. 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.

  18. 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…

  19. 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…

  20. 49 CFR 826.22 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 7 2011-10-01 2011-10-01 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...

  1. 14 CFR 1262.202 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 5 2011-01-01 2010-01-01 true Net worth exhibit. 1262.202 Section 1262.202... PROCEEDINGS Information Required From Applicants § 1262.202 Net worth exhibit. (a) Each applicant except a... exhibit showing the net worth of the applicant and any affiliates (as defiined in § 1262.104(f) when...

  2. 10 CFR 12.202 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 10 Energy 1 2011-01-01 2011-01-01 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 tax-exempt organization or cooperative association must provide with its application a detailed exhibit showing the...

  3. 19 CFR 212.11 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 19 Customs Duties 3 2011-04-01 2011-04-01 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 exhibit... with its application a detailed exhibit showing the net worth of the applicant and any affiliates...

  4. 17 CFR 201.42 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 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 applicant... a detailed exhibit showing the net worth of the applicant and any affiliates (as defined in §...

  5. 49 CFR 6.19 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 1 2011-10-01 2011-10-01 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...

  6. 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...

  7. 31 CFR 6.9 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 31 Money and Finance: Treasury 1 2011-07-01 2011-07-01 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 §...

  8. 7 CFR 1.191 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 1 2011-01-01 2011-01-01 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...

  9. 10 CFR 12.202 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 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 tax-exempt organization or cooperative association must provide with its application a detailed exhibit showing the...

  10. 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...

  11. 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...

  12. 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...

  13. 17 CFR 148.12 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 17 Commodity and Securities Exchanges 1 2011-04-01 2011-04-01 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...

  14. 14 CFR 1262.202 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 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 applicant... detailed exhibit showing the net worth of the applicant and any affiliates (as defiined in §...

  15. 10 CFR 1023.311 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 10 Energy 4 2011-01-01 2011-01-01 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 §...

  16. 17 CFR 201.42 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 17 Commodity and Securities Exchanges 2 2011-04-01 2011-04-01 false Net worth exhibit. 201.42... Regulations Pertaining to the Equal Access to Justice Act § 201.42 Net worth exhibit. (a) Each applicant... a detailed exhibit showing the net worth of the applicant and any affiliates (as defined in §...

  17. 49 CFR 1016.202 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 8 2011-10-01 2011-10-01 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...

  18. 5 CFR 2610.202 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 5 Administrative Personnel 3 2011-01-01 2011-01-01 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...

  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. 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...

  1. 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 §...

  2. 45 CFR 1160.4 - Eligibility for international exhibitions.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 45 Public Welfare 3 2014-10-01 2014-10-01 false Eligibility for international exhibitions. 1160.4... AND ARTIFACTS INDEMNITY ACT § 1160.4 Eligibility for international exhibitions. An indemnity agreement for an international exhibition made under these regulations shall cover: (a) Eligible items...

  3. 45 CFR 1160.4 - Eligibility for international exhibitions.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 45 Public Welfare 3 2011-10-01 2011-10-01 false Eligibility for international exhibitions. 1160.4... AND ARTIFACTS INDEMNITY ACT § 1160.4 Eligibility for international exhibitions. An indemnity agreement for an international exhibition made under these regulations shall cover: (a) Eligible items...

  4. 45 CFR 1160.4 - Eligibility for international exhibitions.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 45 Public Welfare 3 2012-10-01 2012-10-01 false Eligibility for international exhibitions. 1160.4... AND ARTIFACTS INDEMNITY ACT § 1160.4 Eligibility for international exhibitions. An indemnity agreement for an international exhibition made under these regulations shall cover: (a) Eligible items...

  5. 45 CFR 1160.4 - Eligibility for international exhibitions.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 45 Public Welfare 3 2013-10-01 2013-10-01 false Eligibility for international exhibitions. 1160.4... AND ARTIFACTS INDEMNITY ACT § 1160.4 Eligibility for international exhibitions. An indemnity agreement for an international exhibition made under these regulations shall cover: (a) Eligible items...

  6. 45 CFR 1160.4 - Eligibility for international exhibitions.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 45 Public Welfare 3 2010-10-01 2010-10-01 false Eligibility for international exhibitions. 1160.4... AND ARTIFACTS INDEMNITY ACT § 1160.4 Eligibility for international exhibitions. An indemnity agreement for an international exhibition made under these regulations shall cover: (a) Eligible items...

  7. 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,…

  8. 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

  9. 18 CFR 157.16 - Exhibits relating to acquisitions.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ..., Sales, Service, Construction, Extension, Acquisition or Abandonment § 157.16 Exhibits relating to acquisitions. In addition to the exhibits required by § 157.14, every application involving acquisition of... follow the procedures set out in § 157.10(d). (a) Exhibit Q—Effect of acquisition on existing...

  10. 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. PMID:24782807

  11. How to create an effective scientific exhibit: analysis of award-winning exhibits from the 1998 RSNA meeting.

    PubMed

    Choi, J R; Kruskal, J B; Rosen, M P; Davis, R B

    2000-01-01

    Although the most important component of an effective scientific exhibit is content, the way in which an exhibit is constructed can greatly influence its overall effectiveness. Choice of format should be determined by carefully analyzing the purpose of one's exhibit, expected audience, and data at hand, as well as type of meeting and funding. Depending on the type of data to be presented and available equipment and budget, the most appropriate style for a scientific exhibit may be a traditional mat board, computer-generated tiles or large-print backboard panel, traditional mat board with viewbox exhibit, matted transparency tiles with viewbox exhibit, or computer-generated large-film display. The authors analyzed 993 of 1, 041 (95.4%) scientific exhibits on display at the 84th RSNA Scientific Assembly and Annual Meeting and categorized each exhibit according to the following characteristics: display type and size, color scheme, display font size, and graphic styles. These characteristics were then correlated with scientific exhibit and design awards as well as invitations for submission to RadioGraphics. Chance of winning an award or being asked to publish the presentation in RadioGraphics was significantly increased for viewbox exhibits (compared with backboard panel exhibits) and for larger exhibits (compared with smaller exhibits). PMID:10903695

  12. Characterization of two members among the five ADP-forming acyl coenzyme A (Acyl-CoA) synthetases reveals the presence of a 2-(Imidazol-4-yl)acetyl-CoA synthetase in Thermococcus kodakarensis.

    PubMed

    Awano, Tomotsugu; Wilming, Anja; Tomita, Hiroya; Yokooji, Yuusuke; Fukui, Toshiaki; Imanaka, Tadayuki; Atomi, Haruyuki

    2014-01-01

    The genome of Thermococcus kodakarensis, along with those of most Thermococcus and Pyrococcus species, harbors five paralogous genes encoding putative α subunits of nucleoside diphosphate (NDP)-forming acyl coenzyme A (acyl-CoA) synthetases. The substrate specificities of the protein products for three of these paralogs have been clarified through studies on the individual enzymes from Pyrococcus furiosus and T. kodakarensis. Here we have examined the biochemical properties of the remaining two acyl-CoA synthetase proteins from T. kodakarensis. The TK0944 and TK2127 genes encoding the two α subunits were each coexpressed with the β subunit-encoding TK0943 gene. In both cases, soluble proteins with an α2β2 structure were obtained and their activities toward various acids in the ADP-forming reaction were examined. The purified TK0944/TK0943 protein (ACS IIITk) accommodated a broad range of acids that corresponded to those generated in the oxidative metabolism of Ala, Val, Leu, Ile, Met, Phe, and Cys. In contrast, the TK2127/TK0943 protein exhibited relevant levels of activity only toward 2-(imidazol-4-yl)acetate, a metabolite of His degradation, and was thus designated 2-(imidazol-4-yl)acetyl-CoA synthetase (ICSTk), a novel enzyme. Kinetic analyses were performed on both proteins with their respective substrates. In T. kodakarensis, we found that the addition of histidine to the medium led to increases in intracellular ADP-forming 2-(imidazol-4-yl)acetyl-CoA synthetase activity, and 2-(imidazol-4-yl)acetate was detected in the culture medium, suggesting that ICSTk participates in histidine catabolism. The results presented here, together with those of previous studies, have clarified the substrate specificities of all five known NDP-forming acyl-CoA synthetase proteins in the Thermococcales. PMID:24163338

  13. [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.

  14. 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. PMID:22247169

  15. Evolutionary Dynamics of Influenza A Viruses in US Exhibition Swine.

    PubMed

    Nelson, Martha I; Wentworth, David E; Das, Suman R; Sreevatsan, Srinand; Killian, Mary L; Nolting, Jacqueline M; Slemons, Richard D; Bowman, Andrew S

    2016-01-15

    The role of exhibition swine in influenza A virus transmission was recently demonstrated by >300 infections with influenza A(H3N2) variant viruses among individuals who attended agricultural fairs. Through active influenza A virus surveillance in US exhibition swine and whole-genome sequencing of 380 isolates, we demonstrate that exhibition swine are actively involved in the evolution of influenza A viruses, including zoonotic strains. First, frequent introduction of influenza A viruses from commercial swine populations provides new genetic diversity in exhibition pigs each year locally. Second, genomic reassortment between viruses cocirculating in exhibition swine increases viral diversity. Third, viral migration between exhibition swine in neighboring states demonstrates that movements of exhibition pigs contributes to the spread of genetic diversity. The unexpected frequency of viral exchange between commercial and exhibition swine raises questions about the understudied interface between these populations. Overall, the complexity of viral evolution in exhibition swine indicates that novel viruses are likely to continually reemerge, presenting threats to humans. PMID:26243317

  16. 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

  17. 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.

  18. 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

  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. 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…

  1. 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…

  2. 22 CFR Exhibit B to Part 204 - Assignment

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 22 Foreign Relations 1 2010-04-01 2010-04-01 false Assignment B Exhibit B to Part 204 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT HOUSING GUARANTY STANDARD TERMS AND CONDITIONS Pt. 204, Exh. B Exhibit B to Part 204—Assignment The undersigned, being the registered owner of a Note in...

  3. 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......

  4. 17 CFR 229.1016 - (Item 1016) Exhibits.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 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......

  5. 17 CFR 229.1016 - (Item 1016) Exhibits.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 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......

  6. 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…

  7. 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…

  8. 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...

  9. 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...

  10. 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.... A, Exh. F Exhibit F to Subpart A of Part 1924—Payment Bond KNOW ALL PERSONS BY THESE PRESENTS: that... persons, firms, and corporations having a direct contract with the PRINCIPAL or its...

  11. 7 CFR Exhibit F to Subpart A of... - Payment Bond

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 12 2012-01-01 2012-01-01 false Payment Bond F Exhibit F to Subpart A of Part 1924.... A, Exh. F Exhibit F to Subpart A of Part 1924—Payment Bond KNOW ALL PERSONS BY THESE PRESENTS: that... persons, firms, and corporations having a direct contract with the PRINCIPAL or its...

  12. 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.... A, Exh. F Exhibit F to Subpart A of Part 1924—Payment Bond KNOW ALL PERSONS BY THESE PRESENTS: that... persons, firms, and corporations having a direct contract with the PRINCIPAL or its...

  13. 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...

  14. 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...

  15. 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...

  16. 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...

  17. 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...

  18. 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...

  19. 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...

  20. 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...

  1. 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...

  2. 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...

  3. Students-Exhibits Interaction at a Science Center

    ERIC Educational Resources Information Center

    Botelho, Agostinho; Morais, Ana M.

    2006-01-01

    In this study we investigate students' learning during their interaction with two exhibits at a science center. Specifically, we analyze both students' procedures when interacting with exhibits and their understanding of the scientific concepts presented therein. Bernstein's theory of pedagogic discourse (1990, 2000) provided the sociological…

  4. 47 CFR 1.1512 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 47 Telecommunication 1 2011-10-01 2011-10-01 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...

  5. 12 CFR 1203.11 - Confidentiality of net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 12 Banks and Banking 7 2011-01-01 2011-01-01 false Confidentiality of net worth exhibit. 1203.11... INFORMATION ACT Information Required From Applicants § 1203.11 Confidentiality of net worth exhibit. Unless otherwise ordered by the Director, or required by law, the statement of net worth will be for...

  6. 22 CFR 134.12 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 22 Foreign Relations 1 2011-04-01 2011-04-01 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...

  7. 28 CFR 24.202 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 28 Judicial Administration 1 2011-07-01 2011-07-01 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...

  8. 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...

  9. 24 CFR 14.205 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 24 Housing and Urban Development 1 2010-04-01 2010-04-01 false Net worth exhibit. 14.205 Section... Required From Applicants § 14.205 Net worth exhibit. (a) Each applicant except a qualified tax-exempt... the net worth of the applicant and any affiliates (as defined in § 14.120(f) of this part) when...

  10. 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...

  11. 24 CFR 14.205 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 24 Housing and Urban Development 1 2011-04-01 2011-04-01 false Net worth exhibit. 14.205 Section... Required From Applicants § 14.205 Net worth exhibit. (a) Each applicant except a qualified tax-exempt... the net worth of the applicant and any affiliates (as defined in § 14.120(f) of this part) when...

  12. 29 CFR 16.202 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 29 Labor 1 2011-07-01 2011-07-01 false 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... must provide with its application a detailed exhibit showing the net worth of the applicant and...

  13. 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...

  14. 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...

  15. 14 CFR 14.11 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 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...

  16. 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...

  17. 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...

  18. 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...

  19. 32 CFR 705.25 - Navy Exhibit Center.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 32 National Defense 5 2011-07-01 2011-07-01 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...

  20. 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...

  1. 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... 2003, NARA completed a two year renovation of the Rotunda and constructed additional exhibit space...

  2. 17 CFR 229.601 - (Item 601) Exhibits.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... INSTRUCTIONS FOR FILING FORMS UNDER SECURITIES ACT OF 1933, SECURITIES EXCHANGE ACT OF 1934 AND ENERGY POLICY... sequential numbering system where such exhibit can be found. Where exhibits are incorporated by reference.... (iii)(A) Any management contract or any compensatory plan, contract or arrangement, including but...

  3. 17 CFR 229.601 - (Item 601) Exhibits.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... INSTRUCTIONS FOR FILING FORMS UNDER SECURITIES ACT OF 1933, SECURITIES EXCHANGE ACT OF 1934 AND ENERGY POLICY... sequential numbering system where such exhibit can be found. Where exhibits are incorporated by reference.... (iii)(A) Any management contract or any compensatory plan, contract or arrangement, including but...

  4. 7 CFR Exhibit C to Subpart N of... - [Reserved

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 13 2014-01-01 2013-01-01 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...

  5. 7 CFR Exhibit C to Subpart N of... - [Reserved

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 13 2013-01-01 2013-01-01 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...

  6. 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...

  7. Champions of American Sport--A Major Exhibition.

    ERIC Educational Resources Information Center

    Sorani, Bob

    1981-01-01

    An exhibit of 100 photographs, portraits, and sculptures, entitled "Champions of American Sport", is the first major art exhibit ever devoted to outstanding American sports personalities and to the aesthetic qualities of sport and human movement. The Smithsonian Institution is sponsoring the traveling collection, which includes works from the…

  8. 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…

  9. 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...

  10. 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...

  11. 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...

  12. 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...

  13. 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...

  14. 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.…

  15. 7 CFR Exhibit G to Subpart G of... - [Reserved

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 13 2011-01-01 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...

  16. 7 CFR Exhibit G to Subpart A of... - Performance Bond

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 12 2012-01-01 2012-01-01 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...

  17. 7 CFR Exhibit G to Subpart G of... - [Reserved

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 13 2012-01-01 2012-01-01 false 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...

  18. 7 CFR Exhibits A-B to Subpart G... - [Reserved

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 12 2013-01-01 2013-01-01 false A Exhibits A-B to Subpart G to Part 1822 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE..., Procedures, and Authorizations Exhibits A-B to Subpart G to Part 1822...

  19. 7 CFR Exhibit G to Subpart A of... - Performance Bond

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 12 2013-01-01 2013-01-01 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...

  20. 7 CFR Exhibit G to Subpart G of... - [Reserved

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 13 2014-01-01 2013-01-01 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...

  1. 7 CFR Exhibit G to Subpart G of... - [Reserved

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 13 2013-01-01 2013-01-01 false 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...

  2. 7 CFR Exhibit G to Subpart A of... - Performance Bond

    Code of Federal Regulations, 2014 CFR

    2014-01-01

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

  3. 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:...

  4. 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...

  5. 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...

  6. 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...

  7. 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,…

  8. 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...

  9. 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)

  10. 48 CFR 6101.17 - Exhibits [Rule 17].

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 6101.17 Federal Acquisition Regulations System CIVILIAN BOARD OF CONTRACT APPEALS, GENERAL SERVICES ADMINISTRATION CONTRACT DISPUTE CASES 6101.17 Exhibits . (a) Marking of exhibits. (1) Documents and other... electronically stored information or printed versions of electronically stored information to be included in...

  11. 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...

  12. 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...

  13. 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...

  14. 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...

  15. 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...

  16. 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...

  17. 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...

  18. 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....

  19. Three Pictures of an Exhibition: Warm, Cool, and Hard.

    ERIC Educational Resources Information Center

    McDonald, Joseph P.

    1993-01-01

    Describes several ways to view senior exhibits at an urban high school employing the Coalition of Essential Schools'"graduation by exhibition" assessment method. The coalition advocates a pedagogy combining a personalized, caring environment with a focus on student production. Judges must balance warm regard with cool, critical appraisal and…

  20. 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

  1. Cosmic Origins: A Traveling Science Exhibit and Education Program

    NASA Astrophysics Data System (ADS)

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

    2003-12-01

    The Space Science Institute of Boulder, Colorado, is developing a 3,000 square-foot traveling exhibition, called Cosmic Origins, which will bring origins-related research and discoveries to students and the American public. Cosmic Origins will have three interrelated exhibit areas: Star Formation, 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 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. Exhibit content will address age-old questions that form the basis of NASA's Origins and Astrobiology programs: Where did we come from? Are we alone? In addition to the exhibit, our project will include workshops for educators and docents at host sites, as well as a public Web site that will use a virtual rendering of exhibit content. The exhibit's size will permit it to visit medium sized museums in underserved regions of the country. It will begin its 3-year tour to 9 host museums and science centers in early 2005. A second 3-year tour is also planned for 2008. The Association of Science-Technology Centers (ASTC) will manage the exhibit's national tour. Current partners in the Cosmic Origins project include ASTC, the Denver Museum of Nature and Science, Lawrence Hall of Science, NASA Astrobiology Institute, NASA missions (e.g. PlanetQuest, SIRTF, and Kepler), New York Hall of Science, the SETI Institute, and the Space Telescope Science Institute. The exhibition is supported by grants from NSF and NASA. This report will focus on the Planet Quest part of the exhibition.

  2. 7 CFR Exhibit C to Subpart C of... - Checklist of Visual Exhibits and Documentation for RRH, RCH, and LH Proposals

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 12 2011-01-01 2011-01-01 false Checklist of Visual Exhibits and Documentation for RRH, RCH, and LH Proposals C Exhibit C to Subpart C of Part 1924 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE SERVICE, RURAL UTILITIES SERVICE, AND FARM SERVICE...

  3. 7 CFR Exhibit C to Subpart C of... - Checklist of Visual Exhibits and Documentation for RRH, RCH, and LH Proposals

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 12 2010-01-01 2010-01-01 false Checklist of Visual Exhibits and Documentation for RRH, RCH, and LH Proposals C Exhibit C to Subpart C of Part 1924 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE SERVICE, RURAL UTILITIES SERVICE, AND FARM SERVICE...

  4. 7 CFR Exhibit C to Subpart C of... - Checklist of Visual Exhibits and Documentation for RRH, RCH, and LH Proposals

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 12 2013-01-01 2013-01-01 false Checklist of Visual Exhibits and Documentation for RRH, RCH, and LH Proposals C Exhibit C to Subpart C of Part 1924 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE SERVICE, RURAL UTILITIES SERVICE, AND FARM SERVICE...

  5. 7 CFR Exhibit C to Subpart C of... - Checklist of Visual Exhibits and Documentation for RRH, RCH, and LH Proposals

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 12 2012-01-01 2012-01-01 false Checklist of Visual Exhibits and Documentation for RRH, RCH, and LH Proposals C Exhibit C to Subpart C of Part 1924 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE SERVICE, RURAL UTILITIES SERVICE, AND FARM SERVICE...

  6. An exploratory study of zoo visitors' exhibit experiences and reactions.

    PubMed

    Luebke, Jerry F; Matiasek, Jennifer

    2013-01-01

    Visiting a zoo or aquarium is not only fun, but can also have a positive impact on visitors' knowledge and attitudes regarding animals and the environment. The biggest challenge, however, is for these institutions to strategically provide opportunities for cognitive and affective learning while simultaneously facilitating enjoyment and fun. Recent studies in zoos and aquaria have examined various factors that can influence learning such as engaging visitors' emotions or connecting with visitors' prior knowledge and interests. The intent of the current study was to further this line of investigation and explore the relationship between visitors' predispositions and their cognitive and affective experiences and reactions as they walked through an animal exhibit. We selected three indoor immersion exhibits and one outdoor naturalistic exhibit for the study to obtain a wide range of different animals and exhibit settings. Research assistants randomly intercepted visitors leaving the exhibits and asked, among other things, the extent they experienced certain thoughts and feelings while they were walking through the exhibits. Results revealed that visitors' emotional responses to viewing animals were key experiences along with opportunities for introspection and reflection during their time in the exhibits. Implications of the study are discussed in reference to providing both fun and meaningful learning experiences for visitors. PMID:23740472

  7. 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

  8. 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

  9. 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.

  10. 4. PENSTOCKS. EXHIBIT L, SANTA ANA RIVER NO. 1 PROJECT, ...

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

    4. PENSTOCKS. EXHIBIT L, SANTA ANA RIVER NO. 1 PROJECT, APR. 30, 1945. SCE drawing no. 523197 (sheet no. 7; for filing with Federal Power Commission). - Santa Ana River Hydroelectric System, SAR-1 Forebay & Penstock, Redlands, San Bernardino County, CA

  11. 54. ALDER CREEK DIVERSION, PROJECT 1933, EXHIBIT F, SANTA ANA ...

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

    54. ALDER CREEK DIVERSION, PROJECT 1933, EXHIBIT F, SANTA ANA POWERHOUSE NO. 2 SCE drawing no. 5206858, no date (FERC no. 1933-48). - Santa Ana River Hydroelectric System, Redlands, San Bernardino County, CA

  12. 3. TAILRACE AND FOREBAY, SANTA ANA NO. 3, EXHIBIT L, ...

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

    3. TAILRACE AND FOREBAY, SANTA ANA NO. 3, EXHIBIT L, JAN. 25, 1956. SCE drawing no. 541475 (sheet 6; for filing with Federal Power Commission). - Santa Ana River Hydroelectric System, SAR-3 Forebay & Penstock, Redlands, San Bernardino County, CA

  13. 56. CROSS SECTION OF POWERHOUSE, PROJECT 1933, EXHIBIT F, SANTA ...

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

    56. CROSS SECTION OF POWERHOUSE, PROJECT 1933, EXHIBIT F, SANTA ANA POWERHOUSE NO. 1. SCE drawing no. 5206856 (no date; FERC no. 1933-46). - Santa Ana River Hydroelectric System, SAR-1 Powerhouse, Redlands, San Bernardino County, CA

  14. 54. PLAN OF POWERHOUSE, PROJECT 1933, EXHIBIT F, SANTA ANA ...

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

    54. PLAN OF POWERHOUSE, PROJECT 1933, EXHIBIT F, SANTA ANA POWERHOUSE NO. 1. SCE drawing no. 5206855 (no date; FERC no. 1933-45). - Santa Ana River Hydroelectric System, SAR-1 Powerhouse, Redlands, San Bernardino County, CA

  15. "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.

  16. 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. PMID:23286186

  17. 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

  18. The role of exhibits in teacher workshops at science museums

    NASA Astrophysics Data System (ADS)

    Stein, Fred D.

    Between 1986 and 1998, the Exploratorium Institute for Inquiry offered multi-week science institutes for elementary educators involving museum exhibit use during a three-day independent investigation on light and color. Many museums tend to underutilize exhibit use in their teacher education programs. This study addresses the question, "What are the contributions of exhibit use to teachers' learning of science content during a workshop at a science museum?" Data from workshops over three successive years was collected in the form of 13 case studies of participants' investigations. Pre- and post-testing of six participants showed a large (ES = 3.0 SD) and significant gain in their understanding of light and color concepts. The case studies were analyzed by coding each incident of exhibit use according to how the exhibit interaction might have helped the participant in his or her learning. Clusters of recurring themes emerged inductively from the coding process suggesting that the exhibits conferred both logistical and conceptual benefits. Logistically, the exhibits acted as "labor-saving" devices, saving participants time because they were always set up and ready to use, and saving the workshop facilitators time because facilitators could recommend that a participant visit an exhibit rather than spend time giving them individual attention or helping them construct their own investigation apparatus. Conceptually, the exhibits supported each aspect of the Piagetian conceptual change process---accommodation, assimilation, and disequilibrium. They supported accommodation of idea structures and the development of new ones by encouraging participants to ask and answer "What would happen if...?" questions which often generated ideas to explain the new experiences. They supported assimilation of experiences into recently developed idea structures or schemes (supporting and consolidating them) by providing opportunities for participants to ask and answer "Will it happen if

  19. 15 CFR 18.12 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 15 Commerce and Foreign Trade 1 2011-01-01 2011-01-01 false Net worth exhibit. 18.12 Section 18.12... Information Required from Applicants § 18.12 Net worth exhibit. (a) Each applicant except a qualified tax... showing the net worth of the applicant and any affiliates (as defined in § 18.5(f) of this part) when...

  20. 15 CFR 18.12 - Net worth exhibit.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 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 tax... showing the net worth of the applicant and any affiliates (as defined in § 18.5(f) of this part) when...

  1. Bringing Planetary Science to the Public through Traveling Exhibitions

    NASA Astrophysics Data System (ADS)

    Dusenbery, P. B.

    2001-11-01

    The Space Science Institute (SSI) of Boulder, Colorado has recently developed two museum exhibits called the Space Weather Center and MarsQuest. It is currently planning to develop another exhibit called Gas Giants. 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. The Space Weather Center was developed in partnership with various research missions at NASA's Goddard Space Flight Center. The focus of the presentation will be on MarsQuest and Gas Giants. MarsQuest is a 5000 square-foot, 3M, traveling exhibition that is now touring the country. The exhibit's 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 exhibitions 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 Gas Giants, a proposed 4000 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.

  2. 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.

  3. 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.

  4. 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

  5. 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.

  6. The Powerful Educational Potential of Traveling Space Science Exhibits

    NASA Astrophysics Data System (ADS)

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

    2003-04-01

    Five traveling exhibits (both large and small) related to space science are currently touring the U. S., and two more have recently been funded by the National Science Foundation and NASA. These extraordinary educational resources address topics like space weather, Mars exploration, cosmology, the results of Hubble Space Telescope, and the origins of stars, planets, and life. The Space Science Institute (SSI) of Boulder, Colorado, plays leadership roles in four of the seven exhibits. This paper will summarize the nature and itineraries of these exhibits, and how they serve as rallying points for education and public outreach activities across the entire spectrum of science communication. The talk will give special attention to workshops SSI has conducted at MarsQuest host sites for museum educators, docents, and local educators to bolster the host site's ability to do programming around the exhibit content. These workshops have shown promise of leaving a host site with a legacy of new educational capabilities and enhanced connectivity with space scientists and educators in the region. The talk will also address progress on the MarsQuest On-Line project which uses the 5000 sq ft (500 sq m) exhibit as a conceptual framework for an interactive website.

  7. 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.

  8. 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.

  9. Mouse embryonic fibroblasts exhibit extensive developmental and phenotypic diversity

    PubMed Central

    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

  10. 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

  11. 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

  12. Meeting Report: Ordinary Meeting and Exhibition Meeting, 2006 June 24

    NASA Astrophysics Data System (ADS)

    McGee, H.; Mobberley, M.

    2007-06-01

    Prior to the Ordinary Meeting, the annual Exhibition was opened by Richard Miles, President, who said he was pleased to report that all but two of the Sections had display stands this year, and that most of the Directors had also been able to attend and were available for members who wished to discuss their work. Although we had almost a full day of talks to enjoy, members should not be shy of coming and going from the lecture theatre at will, and in particular must make sure they took sufficient time to do justice to the excellent Exhibition on offer.

  13. Bacterial start site prediction.

    PubMed

    Hannenhalli, S S; Hayes, W S; Hatzigeorgiou, A G; Fickett, J W

    1999-09-01

    With the growing number of completely sequenced bacterial genes, accurate gene prediction in bacterial genomes remains an important problem. Although the existing tools predict genes in bacterial genomes with high overall accuracy, their ability to pinpoint the translation start site remains unsatisfactory. In this paper, we present a novel approach to bacterial start site prediction that takes into account multiple features of a potential start site, viz., ribosome binding site (RBS) binding energy, distance of the RBS from the start codon, distance from the beginning of the maximal ORF to the start codon, the start codon itself and the coding/non-coding potential around the start site. Mixed integer programing was used to optimize the discriminatory system. The accuracy of this approach is up to 90%, compared to 70%, using the most common tools in fully automated mode (that is, without expert human post-processing of results). The approach is evaluated using Bacillus subtilis, Escherichia coli and Pyrococcus furiosus. These three genomes cover a broad spectrum of bacterial genomes, since B.subtilis is a Gram-positive bacterium, E.coli is a Gram-negative bacterium and P. furiosus is an archaebacterium. A significant problem is generating a set of 'true' start sites for algorithm training, in the absence of experimental work. We found that sequence conservation between P. furiosus and the related Pyrococcus horikoshii clearly delimited the gene start in many cases, providing a sufficient training set. PMID:10446249

  14. 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…

  15. 2. SPILLWAYS AND ROCKDROP, SANTA ANA NO. 3, EXHIBIT L, ...

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

    2. SPILLWAYS AND ROCK-DROP, SANTA ANA NO. 3, EXHIBIT L, JAN. 25, 1956. SCE drawing no. 541724 (sheet 5; for filing with Federal Power Commission). - Santa Ana River Hydroelectric System, SAR-3 Forebay & Penstock, Redlands, San Bernardino County, CA

  16. 43. FLOOR PLAN OF POWER HOUSE, EXHIBIT L, SANTA ANA ...

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

    43. FLOOR PLAN OF POWER HOUSE, EXHIBIT L, SANTA ANA RIVER NO. 2 PROJECT, APR. 30, 1945. SCE drawing no. 523643 (sheet no. 14; for filing with Federal Power Commission). - Santa Ana River Hydroelectric System, SAR-2 Powerhouse, Redlands, San Bernardino County, CA

  17. 10. DETAILS OF STEEL FLUME, TYPICAL BENTS AND TRUSSES. EXHIBIT ...

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

    10. DETAILS OF STEEL FLUME, TYPICAL BENTS AND TRUSSES. EXHIBIT L, SANTA ANA RIVER NO. 1 PROJECT, APR. 30, 1945. SCE drawing no. 523196 (sheet no. 6; for filing with Federal Power Commission). - Santa Ana River Hydroelectric System, Flumes & Tunnels below Sandbox, Redlands, San Bernardino County, CA

  18. 4. FOREBAY AND PENSTOCK, EXHIBIT L, SANTA ANA RIVER NO. ...

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

    4. FOREBAY AND PENSTOCK, EXHIBIT L, SANTA ANA RIVER NO. 2 PROJECT, APR. 30, 1945. SCE drawing no. 523642 (sheet no. 13; for filing with the Federal Power Commission). - Santa Ana River Hydroelectric System, SAR-2 Forebay & Penstock, Redlands, San Bernardino County, CA

  19. 9. DETAILS OF STEEL FLUME, TYPICAL CURVES AND TRANSITIONS. EXHIBIT ...

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

    9. DETAILS OF STEEL FLUME, TYPICAL CURVES AND TRANSITIONS. EXHIBIT L, SANTA ANA RIVER NO. 1 PROJECT, APR. 30, 1945. SCE drawing no. 523195 (sheet no. 5; for filing with Federal Power Commission). - Santa Ana River Hydroelectric System, Flumes & Tunnels below Sandbox, Redlands, San Bernardino County, CA

  20. 9. HIGH LENNON FLUME, SANTA ANA NO 3, EXHIBIT L, ...

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

    9. HIGH LENNON FLUME, SANTA ANA NO 3, EXHIBIT L, JAN. 25, 1956. SCE drawing no. 541723 (sheet 3; for filing with Federal Power Commission). - Santa Ana River Hydroelectric System, Warm Springs Canyon-SAR-3 Flumes, Redlands, San Bernardino County, CA

  1. 5. SANDBOX BETWEEN TUNNELS 12. SANTA ANA NO. 3, EXHIBIT ...

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

    5. SANDBOX BETWEEN TUNNELS 1-2. SANTA ANA NO. 3, EXHIBIT L, JAN. 25, 1956. SCE drawing no. 541727 (sheet 2; for filing with Federal Power Commission). - Santa Ana River Hydroelectric System, Sandbox, SAR-3 Flowline, Redlands, San Bernardino County, CA

  2. 55. CROSS SECTION OF POWER HOUSE, EXHIBIT L, SANTA ANA ...

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

    55. CROSS SECTION OF POWER HOUSE, EXHIBIT L, SANTA ANA RIVER NO. 1 PROJECT, APR. 30, 1945. SCE drawing no. 523199 (sheet no. 9, for filing with Federal Power Commission). - Santa Ana River Hydroelectric System, SAR-1 Powerhouse, Redlands, San Bernardino County, CA

  3. 44. SECTIONS OF POWER HOUSE, EXHIBIT L, SANTA ANA RIVER ...

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

    44. SECTIONS OF POWER HOUSE, EXHIBIT L, SANTA ANA RIVER NO. 2 PROJECT, APR. 30, 1945. SCE drawing no. 523644 (sheet no. 15; for filing with Federal Power Commission). - Santa Ana River Hydroelectric System, SAR-2 Powerhouse, Redlands, San Bernardino County, CA

  4. 7 CFR Exhibit B to Subpart B of... - Servicing Company

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... of Agriculture, 7 CFR 2.23; delegation of authority by the Assistant Secretary for Rural Development, 7 CFR 2.70) ... 7 Agriculture 12 2011-01-01 2011-01-01 false Servicing Company B Exhibit B to Subpart B of...

  5. 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...

  6. 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

  7. 12 CFR 625.11 - Net worth exhibit.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... regarding release of information (12 CFR part 602). ... 12 Banks and Banking 6 2011-01-01 2011-01-01 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...

  8. 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...

  9. 12 CFR 625.11 - Net worth exhibit.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... regarding release of information (12 CFR part 602). ... 12 Banks and Banking 7 2013-01-01 2013-01-01 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...

  10. 12 CFR 625.11 - Net worth exhibit.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... regarding release of information (12 CFR part 602). ... 12 Banks and Banking 7 2012-01-01 2012-01-01 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. 12 CFR 625.11 - Net worth exhibit.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... regarding release of information (12 CFR part 602). ... 12 Banks and Banking 7 2014-01-01 2014-01-01 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...

  12. A Preliminary Evaluation of the "Birds in Canada" Exhibit.

    ERIC Educational Resources Information Center

    Clowes, Marya; Wolff, Lee

    This study of the Birds in Canada exhibit (National Museum of Natural Sciences) examines the effectiveness of traditional and modern displays and the visitors' use of identification labels, text, map, and film. Over 700 museum visitors comprised the random sample. Evaluation methods included observation, interview, and cognitive testing, using…

  13. 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...

  14. 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...

  15. 7 CFR Appendix - Exhibits to Subpart E of Part 1951

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 14 2013-01-01 2013-01-01 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...

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

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 12 2011-01-01 2011-01-01 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-COOPERATIVE SERVICE, RURAL UTILITIES SERVICE, AND FARM SERVICE AGENCY, DEPARTMENT OF AGRICULTURE PROGRAM REGULATIONS CONSTRUCTION AND...

  17. 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 of the applicant and any affiliates (as defined in § 1016.105(f) of this part) when the...

  18. 14 CFR 77.59 - Subpoenas of witnesses and exhibits.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 2 2011-01-01 2011-01-01 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 (Eff. until 1-18-11) Rules of Practice for...

  19. 17 CFR 229.1016 - (Item 1016) Exhibits.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 17 Commodity and Securities Exchanges 2 2012-04-01 2012-04-01 false (Item 1016) Exhibits. 229.1016... AND CONSERVATION ACT OF 1975-REGULATION S-K Mergers and Acquisitions (Regulation M-A) § 229.1016 (Item... letter); (2) Solicitation or recommendation (including those referred to in Item 1012 of Regulation...

  20. 17 CFR 229.1016 - (Item 1016) Exhibits.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 17 Commodity and Securities Exchanges 2 2011-04-01 2011-04-01 false (Item 1016) Exhibits. 229.1016... AND CONSERVATION ACT OF 1975-REGULATION S-K Mergers and Acquisitions (Regulation M-A) § 229.1016 (Item... letter); (2) Solicitation or recommendation (including those referred to in Item 1012 of Regulation...

  1. 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…

  2. Grandma Moses in the 21st Century. Learning from Exhibitions.

    ERIC Educational Resources Information Center

    Johnson, Mark M.

    2001-01-01

    Provides background information on the life and career of Grandma Moses who was born as Anna Mary Robertson and painted in the style of folk or naive art. Addresses the art exhibition entitled "Grandma Moses in the 21st Century" that explores the recurring themes in her artwork. (CMK)

  3. "Creation's Journey": Excerpts from the Inaugural Exhibition and Book.

    ERIC Educational Resources Information Center

    Hill, Tom, Hill, Richard W., Sr., Ed.

    1994-01-01

    Contains excerpts from the companion book to the inaugural exhibition "Creation's Journey: Masterworks of Native American Identity and Belief" at the George Gustav Heye Center of the National Museum of the American Indian in New York City. Describes the cultural traditions behind some of the 165 objects chosen for their artistic achievement,…

  4. 27 CFR 4.51 - Exhibiting certificates to Government officials.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Exhibiting certificates to Government officials. 4.51 Section 4.51 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF WINE Requirements...

  5. 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,…

  6. 17 CFR 148.12 - Net worth exhibit.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... to the Presiding Officer in a sealed envelope labeled “Confidential Financial Information... 17 Commodity and Securities Exchanges 1 2013-04-01 2013-04-01 false Net worth exhibit. 148.12 Section 148.12 Commodity and Securities Exchanges COMMODITY FUTURES TRADING COMMISSION IMPLEMENTATION...

  7. 17 CFR 148.12 - Net worth exhibit.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... directly to the Presiding Officer in a sealed envelope labeled “Confidential Financial Information... 17 Commodity and Securities Exchanges 2 2014-04-01 2014-04-01 false Net worth exhibit. 148.12 Section 148.12 Commodity and Securities Exchanges COMMODITY FUTURES TRADING COMMISSION...

  8. 17 CFR 148.12 - Net worth exhibit.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... to the Presiding Officer in a sealed envelope labeled “Confidential Financial Information... 17 Commodity and Securities Exchanges 1 2012-04-01 2012-04-01 false Net worth exhibit. 148.12 Section 148.12 Commodity and Securities Exchanges COMMODITY FUTURES TRADING COMMISSION IMPLEMENTATION...

  9. 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…

  10. 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...

  11. 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...

  12. 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 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...

  13. 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...

  14. 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...

  15. Computerizing Museum Exhibits: The Beginnings of the Essential Conversation

    ERIC Educational Resources Information Center

    Burdick, Bruce

    1978-01-01

    Examples are given of some types of interaction between viewed and museum provided by an experimental computer based nutrition exhibit at the Chicago Museum of Science and Industry. Plans are being developed for a museum in which most of the information conveyed will appear through the use of video systems tied to a computer. (CMV)

  16. 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

  17. 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…

  18. 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...

  19. 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...

  20. 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...

  1. 45 CFR 1160.5 - Eligibility for domestic exhibitions.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 45 Public Welfare 3 2010-10-01 2010-10-01 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...

  2. 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...

  3. 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

  4. 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

  5. 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.

  6. 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

  7. 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...

  8. 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...

  9. (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…

  10. New Viruses Identified in Fig Trees Exhibiting Fig Mosaic Disease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fig mosaic disease has been known for decades, but the causal agent has been elusive. Here we present data on the incidence of at least four new viruses isolated from fig trees exhibiting mosaic symptoms. One of the viruses is closely related to the recently identified European mountain ash ringspo...

  11. 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 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION PROCEDURAL RULES RULES IMPLEMENTING THE EQUAL ACCESS TO JUSTICE ACT OF 1980 Information Required From Applicants § 14.11 Net...

  12. 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…

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

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE SERVICE, RURAL UTILITIES SERVICE, AND FARM SERVICE AGENCY, DEPARTMENT OF AGRICULTURE PROGRAM REGULATIONS CONSTRUCTION AND REPAIR Planning and Performing Site Development Work Exhibit A to Subpart C of Part 1924...

  14. 7 CFR Exhibit A to Subpart C 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 C of Part 1924 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 PROGRAM REGULATIONS CONSTRUCTION AND REPAIR Planning...

  15. 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

  16. 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. 

  17. 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

  18. 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

  19. 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. PMID:22145669

  20. 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)