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Sample records for comamonas denitrificans based

  1. Characterization of Comamonas thiooxidans sp. nov., and comparison of thiosulfate oxidation with Comamonas testosteroni and Comamonas composti.

    PubMed

    Narayan, Kunwar Digvijay; Pandey, Shachindra K; Das, Subrata K

    2010-10-01

    Comamonas thiooxidans (strain S23(T)) capable of oxidizing thiosulfate under a mixotrophic growth condition was isolated from a sulfur spring. DNA-DNA homology study showed 55% similarity with Comamonas testosteroni KCTC2990(T) and 52% with Comamonas composti LMG24008(T), the nearest phylogenetic relative (16S rRNA sequence similarity <97%). Comparative genomic fingerprinting by using ERIC and Rep-PCR further delineated species identity of the strain S23(T) for which Comamonas thiooxidans sp. nov. is proposed. In addition, thiosulfate oxidation potential of the strain S23(T) was compared with Comamonas testosteroni and Comamonas composti.

  2. Elevated level of the second messenger c-di-GMP in Comamonas testosteroni enhances biofilm formation and biofilm-based biodegradation of 3-chloroaniline.

    PubMed

    Wu, Yichao; Ding, Yuanzhao; Cohen, Yehuda; Cao, Bin

    2015-02-01

    The bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP) is a ubiquitous second messenger that determines bacterial lifestyle between the planktonic and biofilm modes of life. Although the role of c-di-GMP signaling in biofilm development and dispersal has been extensively studied, how c-di-GMP signaling influences environmental bioprocess activities such as biodegradation remains unexplored. To elucidate the impacts of elevating c-di-GMP level on environmental bioprocesses, we constructed a Comamonas testosteroni strain constitutively expressing a c-di-GMP synthase YedQ from Escherichia coli and examined its capability in biofilm formation and biodegradation of 3-chloroaniline (3-CA). The high c-di-GMP strain exhibited an increased binding to Congo red dye, a decreased motility, and an enhanced biofilm formation capability. In planktonic cultures, the strain with an elevated c-di-GMP concentration and the wild type could degrade 3-CA comparably well. However, under batch growth conditions with a high surface to volume ratio, an elevated c-di-GMP concentration in C. testosteroni significantly increased the contribution of biofilms in 3-CA biodegradation. In continuous submerged biofilm reactors, C. testosteroni with an elevated c-di-GMP level exhibited an enhanced 3-CA biodegradation and a decreased cell detachment rate. Taken together, this study provides a novel strategy to enhance biofilm-based biodegradation of toxic xenobiotic compounds through manipulating bacterial c-di-GMP signaling.

  3. The 5S ribosomal RNAs of Paracoccus denitrificans and Prochloron

    NASA Technical Reports Server (NTRS)

    Mackay, R. M.; Salgado, D.; Bonen, L.; Doolittle, W. F.; Stackebrandt, E.

    1982-01-01

    The nucleotide sequences of the 5S rRNAs of Paracoccus denitrificans and Prochloron sp. are presented, along with the demonstrated phylogenetic relationships of P. denitrificans with purple nonsulfur bacteria, and of Prochloron with cyanobacteria. Structural findings include the following: (1) helix II in both models is much shorter than in other eubacteria, (2) a base-pair has been deleted from helix IV of P. denitrificans 5S, and (3) Prochloron 5S has the potential to form four base-pairs between residues. Also covered are the differences between pairs of sequences in P. denitrificans, Prochloron, wheat mitochondion, spinach chloroplast, and nine diverse eubacteria. Findings include the observation that Prochloron 5S rRNA is much more similar to the 5S of the cyanobacterium Anacystis nidulans (25 percent difference) than either are to any of the other nine eubacterial 5S rRNAs.

  4. Draft Genome Sequence of Comamonas thiooxydans Strain S23T (DSM 17888T), a Thiosulfate-Oxidizing Bacterium Isolated from a Sulfur Spring in India

    PubMed Central

    Narayan, Kunwar Digvijay; Badhai, Jhasketan; Whitman, William B.

    2016-01-01

    The genus Comamonas contains species isolated from various environments, such as termite guts, wetlands, activated sludge, soil, humans, and fresh water. Here, we report the draft genome sequence of Comamonas thiooxydans strain S23T capable of oxidizing thiosulfate under mixotrophic growth conditions. Based upon draft genome sequencing, the genome is 5.3 Mb and encodes 4,767 proteins. The Comamonas thiooxydans whole-genome sequence will help understand the metabolic diversity in sulfur oxidation pathways. PMID:27516520

  5. Draft Genome Sequence of Comamonas thiooxydans Strain S23T (DSM 17888T), a Thiosulfate-Oxidizing Bacterium Isolated from a Sulfur Spring in India.

    PubMed

    Narayan, Kunwar Digvijay; Badhai, Jhasketan; Whitman, William B; Das, Subrata K

    2016-01-01

    The genus Comamonas contains species isolated from various environments, such as termite guts, wetlands, activated sludge, soil, humans, and fresh water. Here, we report the draft genome sequence of Comamonas thiooxydans strain S23(T) capable of oxidizing thiosulfate under mixotrophic growth conditions. Based upon draft genome sequencing, the genome is 5.3 Mb and encodes 4,767 proteins. The Comamonas thiooxydans whole-genome sequence will help understand the metabolic diversity in sulfur oxidation pathways. PMID:27516520

  6. Intra-Abdominal Infections Due to Comamonas kerstersii

    PubMed Central

    Almuzara, Marisa N.; Cittadini, Rosana; Vera Ocampo, Cecilia; Bakai, Romina; Traglia, German; Ramirez, Maria S.; del Castillo, Marcelo

    2013-01-01

    Herein, we report four cases of Comamonas kerstersii intra-abdominal infections representing the first report of human infections caused by this Comamonas species. In addition, our work demonstrates the association of C. kerstersii with peritonitis secondary to appendix rupture. PMID:23576541

  7. Oxidation of hydrogen sulfide by mixed cultures of Thiobacillus denitrificans and heterotrophs

    SciTech Connect

    Sublette, K.L.; Sylvester, N.D.

    1987-04-01

    A process for the microbial desulfurization of natural gas based on the oxidation of H/sub 2/S(g) by Thiobacillus denitrificans has been previously proposed. The proposed process could be greatly simplified if aseptic operation of the reactor was not required. Accordingly, the authors report here a study of the effects of heterotrophic contaminants on H/sub 2/S(g) oxidation by T. denitrificans.

  8. Genetic manipulation of the obligate chemolithoautotrophic bacterium Thiobacillus denitrificans

    SciTech Connect

    Beller, H.R.; Legler, T.C.; Kane, S.R.

    2011-07-15

    Chemolithoautotrophic bacteria can be of industrial and environmental importance, but they present a challenge for systems biology studies, as their central metabolism deviates from that of model organisms and there is a much less extensive experimental basis for their gene annotation than for typical organoheterotrophs. For microbes with sequenced genomes but unconventional metabolism, the ability to create knockout mutations can be a powerful tool for functional genomics and thereby render an organism more amenable to systems biology approaches. In this chapter, we describe a genetic system for Thiobacillus denitrificans, with which insertion mutations can be introduced by homologous recombination and complemented in trans. Insertion mutations are generated by in vitro transposition, the mutated genes are amplified by the PCR, and the amplicons are introduced into T. denitrificans by electroporation. Use of a complementation vector, pTL2, based on the IncP plasmid pRR10 is also addressed.

  9. Comparison of Four Comamonas Catabolic Plasmids Reveals the Evolution of pBHB To Catabolize Haloaromatics

    PubMed Central

    Chen, Kai; Xu, Xihui; Zhang, Long; Gou, Zhenjiu; Li, Shunpeng

    2015-01-01

    Comamonas plasmids play important roles in shaping the phenotypes of their hosts and the adaptation of these hosts to changing environments, and understanding the evolutionary strategy of these plasmids is thus of great concern. In this study, the sequence of the 119-kb 3,5-dibromo-4-hydroxybenzonitrile-catabolizing plasmid pBHB from Comamonas sp. strain 7D-2 was studied and compared with those of three other Comamonas haloaromatic catabolic plasmids. Incompatibility group determination based on a phylogenetic analysis of 24 backbone gene proteins, as well as TrfA, revealed that these four plasmids all belong to the IncP-1β subgroup. Comparison of the four plasmids revealed a conserved backbone region and diverse genetic-load regions. The four plasmids share a core genome consisting of 40 genes (>50% similarities) and contain 12 to 50 unique genes each, most of which are xenobiotic-catabolic genes. Two functional reductive dehalogenase gene clusters are specifically located on pBHB, showing distinctive evolution of pBHB for haloaromatics. The higher catabolic ability of the bhbA2B2 cluster than the bhbAB cluster may be due to the transcription levels and the character of the dehalogenase gene itself rather than that of its extracytoplasmic binding receptor gene. The plasmid pBHB is riddled with transposons and insertion sequence (IS) elements, and ISs play important roles in the evolution of pBHB. The analysis of the origin of the bhb genes on pBHB suggested that these accessory genes evolved independently. Our work provides insights into the evolutionary strategies of Comamonas plasmids, especially into the adaptation mechanism employed by pBHB for haloaromatics. PMID:26682859

  10. Comparison of Four Comamonas Catabolic Plasmids Reveals the Evolution of pBHB To Catabolize Haloaromatics.

    PubMed

    Chen, Kai; Xu, Xihui; Zhang, Long; Gou, Zhenjiu; Li, Shunpeng; Freilich, Shiri; Jiang, Jiandong

    2015-12-18

    Comamonas plasmids play important roles in shaping the phenotypes of their hosts and the adaptation of these hosts to changing environments, and understanding the evolutionary strategy of these plasmids is thus of great concern. In this study, the sequence of the 119-kb 3,5-dibromo-4-hydroxybenzonitrile-catabolizing plasmid pBHB from Comamonas sp. strain 7D-2 was studied and compared with those of three other Comamonas haloaromatic catabolic plasmids. Incompatibility group determination based on a phylogenetic analysis of 24 backbone gene proteins, as well as TrfA, revealed that these four plasmids all belong to the IncP-1β subgroup. Comparison of the four plasmids revealed a conserved backbone region and diverse genetic-load regions. The four plasmids share a core genome consisting of 40 genes (>50% similarities) and contain 12 to 50 unique genes each, most of which are xenobiotic-catabolic genes. Two functional reductive dehalogenase gene clusters are specifically located on pBHB, showing distinctive evolution of pBHB for haloaromatics. The higher catabolic ability of the bhbA2B2 cluster than the bhbAB cluster may be due to the transcription levels and the character of the dehalogenase gene itself rather than that of its extracytoplasmic binding receptor gene. The plasmid pBHB is riddled with transposons and insertion sequence (IS) elements, and ISs play important roles in the evolution of pBHB. The analysis of the origin of the bhb genes on pBHB suggested that these accessory genes evolved independently. Our work provides insights into the evolutionary strategies of Comamonas plasmids, especially into the adaptation mechanism employed by pBHB for haloaromatics.

  11. Identification of type 4 pili in Kingella denitrificans.

    PubMed Central

    Weir, S; Marrs, C F

    1992-01-01

    Kingella denitrificans is an occasionally pathogenic member of the family Neisseriaceae and is a member of the normal respiratory flora. Electron microscopy, colony morphology types, DNA transformation patterns, and immunoblots suggest that K. denitrificans and K. kingae have type 4 pili. This was confirmed by N-terminal amino acid sequencing for K. denitrificans. Images PMID:1353484

  12. Transfer of Halobacterium denitrificans (Tomlinson, Jahnke, and Hochstein) to the genus Haloferax as Haloferax denitrificans comb. nov

    NASA Technical Reports Server (NTRS)

    Tindall, B. J.; Tomlinson, G. A.; Hochstein, L. I.

    1989-01-01

    Halobacterium denitrificans (Tomlinson, Jahnke, and Hochstein) was described at a time when the taxonomic subdivision of the family Halobacteriaceae was in a state of flux. On the basis of both biochemical and chemotaxonomic data, this organism exhibits features which indicate that it is more closely related to members of the genus Haloferax. On the basis of such criteria, we propose that Halobacterium denitrificans be reclassified as Haloferax denitrificans comb. nov. The type strain is strain ATCC 35960 (= DSM 4425).

  13. Assimilation of ammonia in Paracoccus denitrificans.

    PubMed

    Mikes, V; Chválová, H; Mátlová, L

    1991-01-01

    Two pathways serve for assimilation of ammonia in Paracoccus denitrificans. Glutamate dehydrogenase (NADP+) catalyzes the assimilation at a high NH4+ concentration. If nitrate serves as the nitrogen source, glutamate is synthesized by glutamate-ammonia ligase and glutamate synthase (NADPH). At a very low NH4+ concentration, all three enzymes are synthesized simultaneously. No direct relationship exists between glutamate dehydrogenase (NADP+) and glutamate-ammonia ligase in P. denitrificans, while the glutamate synthase (NADPH) activity changes in parallel with that of the latter enzyme. Ammonia does not influence the induction or repression of glutamate dehydrogenase (NADP+). The inner concentration of metabolites indicates a possible repression of glutamate dehydrogenase (NADP+) by the high concentration of glutamine or its metabolic products as in the case when NH4+ is formed by assimilative nitrate reduction. No direct effect of the intermediates of nitrate assimilation on the synthesis of glutamate dehydrogenase (NADP+) was observed. PMID:1688163

  14. Anoxic Androgen Degradation by the Denitrifying Bacterium Sterolibacterium denitrificans via the 2,3-seco Pathway

    PubMed Central

    Wang, Po-Hsiang; Yu, Chang-Ping; Lee, Tzong-Huei; Lin, Ching-Wen; Ismail, Wael; Wey, Shiaw-Pyng; Kuo, An-Ti

    2014-01-01

    The biodegradation of steroids is a crucial biochemical process mediated exclusively by bacteria. So far, information concerning the anoxic catabolic pathways of androgens is largely unknown, which has prevented many environmental investigations. In this work, we show that Sterolibacterium denitrificans DSMZ 13999 can anaerobically mineralize testosterone and some C19 androgens. By using a 13C-metabolomics approach and monitoring the sequential appearance of the intermediates, we demonstrated that S. denitrificans uses the 2,3-seco pathway to degrade testosterone under anoxic conditions. Furthermore, based on the identification of a C17 intermediate, we propose that the A-ring cleavage may be followed by the removal of a C2 side chain at C-5 of 17-hydroxy-1-oxo-2,3-seco-androstan-3-oic acid (the A-ring cleavage product) via retro-aldol reaction. The androgenic activities of the bacterial culture and the identified intermediates were assessed using the lacZ-based yeast androgen assay. The androgenic activity in the testosterone-grown S. denitrificans culture decreased significantly over time, indicating its ability to eliminate androgens. The A-ring cleavage intermediate (≤500 μM) did not exhibit androgenic activity, whereas the sterane-containing intermediates did. So far, only two androgen-degrading anaerobes (Sterolibacterium denitrificans DSMZ 13999 [a betaproteobacterium] and Steroidobacter denitrificans DSMZ 18526 [a gammaproteobacterium]) have been isolated and characterized, and both of them use the 2,3-seco pathway to anaerobically degrade androgens. The key intermediate 2,3-seco-androstan-3-oic acid can be used as a signature intermediate for culture-independent environmental investigations of anaerobic degradation of C19 androgens. PMID:24657867

  15. The 5S ribosomal RNAs of Paracoccus denitrificans and Prochloron.

    PubMed Central

    MacKay, R M; Salgado, D; Bonen, L; Stackebrandt, E; Doolittle, W F

    1982-01-01

    The nucleotide sequences of the 5S rRNAs of Paracoccus denitrificans and Prochloron sp. are (formula: see text), respectively. Specific phylogenetic relationships of P. denitrificans with purple non-sulphur bacteria, and of Prochloron with cyanobacteria are demonstrated, and unique features of potential secondary structure are described. PMID:7099971

  16. Permanent draft genome sequence of Comamonas testosteroni KF-1

    SciTech Connect

    Weiss, Michael; Kesberg, Anna I; LaButti, Kurt; Pitluck, Sam; Bruce, David; Hauser, Loren John; Copeland, A; Woyke, Tanja; Lowry, Stephen; Lucas, Susan; Land, Miriam L; Goodwin, Lynne A.; Kjelleberg, Staffan; Cook, Alasdair M.; Buhmann, Matthias; Thomas, Torsten; Schleheck, David

    2013-01-01

    Comamonas testosteroni KF-1 is a model organism for the elucidation of the novel biochemical degra- dation pathways for xenobiotic 4-sulfophenylcarboxylates (SPC) formed during biodegradation of syn- thetic 4-sulfophenylalkane surfactants (linear alkylbenzenesulfonates, LAS) by bacterial communities. Here we describe the features of this organism, together with the complete genome sequence and an- notation. The 6,026,527 bp long chromosome (one sequencing gap) exhibits an average G+C content of 61.79% and is predicted to encode 5,492 protein-coding genes and 114 RNA genes.

  17. Permanent draft genome sequence of Comamonas testosteroni KF-1

    PubMed Central

    Weiss, Michael; Kesberg, Anna I.; LaButti, Kurt M.; Pitluck, Sam; Bruce, David; Hauser, Loren; Copeland, Alex; Woyke, Tanja; Lowry, Stephen; Lucas, Susan; Land, Miriam; Goodwin, Lynne; Kjelleberg, Staffan; Cook, Alasdair M.; Buhmann, Matthias; Thomas, Torsten; Schleheck, David

    2013-01-01

    Comamonas testosteroni KF-1 is a model organism for the elucidation of the novel biochemical degradation pathways for xenobiotic 4-sulfophenylcarboxylates (SPC) formed during biodegradation of synthetic 4-sulfophenylalkane surfactants (linear alkylbenzenesulfonates, LAS) by bacterial communities. Here we describe the features of this organism, together with the complete genome sequence and annotation. The 6,026,527 bp long chromosome (one sequencing gap) exhibits an average G+C content of 61.79% and is predicted to encode 5,492 protein-coding genes and 114 RNA genes. PMID:23991256

  18. Genome of the epsilonproteobacterial chemolithoautotroph Sulfurimonas denitrificans

    SciTech Connect

    Sievert, Stefan M; Scott, Kathleen M; Klotz, Martin G; Chain, Patrick S. G.; Hauser, Loren John; Hemp, James; Hugler, Michael; Land, Miriam L; Lapidus, Alla L.; Larimer, Frank W; Lucas, Susan; Malfatti, Stephanie; Meyer, Folker; Paulsen, Ian T; Ren, Qinghu; Simon, Jorg

    2008-01-01

    Sulfur-oxidizing epsilonproteobacteria are common in a variety of sulfidogenic environments. These autotrophic and mixotrophic sulfur-oxidizing bacteria are believed to contribute substantially to the oxidative portion of the global sulfur cycle. In order to better understand the ecology and roles of sulfur-oxidizing epsilonproteobacteria, in particular those of the widespread genus Sulfurimonas, in biogeochemical cycles, the genome of Sulfurimonas denitrificans DSM1251 was sequenced. This genome has many features, including a larger size (2.2 Mbp), that suggest a greater degree of metabolic versatility or responsiveness to the environment than seen for most of the other sequenced epsilonproteobacteria. A branched electron transport chain is apparent, with genes encoding complexes for the oxidation of hydrogen, reduced sulfur compounds, and formate and the reduction of nitrate and oxygen. Genes are present for a complete, autotrophic reductive citric acid cycle. Many genes are present that could facilitate growth in the spatially and temporally heterogeneous sediment habitat from where Sulfurimonas denitrificans was originally isolated. Many resistance-nodulation-development family transporter genes (10 total) are present; of these, several are predicted to encode heavy metal efflux transporters. An elaborate arsenal of sensory and regulatory protein-encoding genes is in place, as are genes necessary to prevent and respond to oxidative stress.

  19. Microbial degradation of quinoline: Kinetic studies with Comamonas acidovorans DSM 6426

    SciTech Connect

    Miethling, R.; Hecht, V.; Deckwer, W.D. . Dept. of Biochemical Engineering)

    1993-08-20

    The microbial degradation of quinoline by Comamonas acidovorans was studied in a laboratory scale stirred tank reactor. In continuous culture experiments using quinoline as a sole source of carbon and nitrogen, it was shown by means of mass balances that quinoline was converted completely to biomass, carbon dioxide, and ammonia. Degradation rates up to 0.7 g/L h were obtained. Measured yield coefficients Y[sub x/s] for quinoline were about 0.7 g/g, which is in agreement with the theoretical value for complete mineralization. Kinetic constants based on Haldane substrate inhibition were evaluated. The values were [mu][sub max] = 0.48 h[sup [minus]1], K[sub i] = 69 mg/L, and K[sub s]<1.45 mg/L.

  20. Identification of Comamonas testosteroni as an androgen degrader in sewage

    PubMed Central

    Chen, Yi-Lung; Wang, Chia-Hsiang; Yang, Fu-Chun; Ismail, Wael; Wang, Po-Hsiang; Shih, Chao-Jen; Wu, Yu-Ching; Chiang, Yin-Ru

    2016-01-01

    Numerous studies have reported the masculinization of freshwater wildlife exposed to androgens in polluted rivers. Microbial degradation is a crucial mechanism for eliminating steroid hormones from contaminated ecosystems. The aerobic degradation of testosterone was observed in various bacterial isolates. However, the ecophysiological relevance of androgen-degrading microorganisms in the environment is unclear. Here, we investigated the biochemical mechanisms and corresponding microorganisms of androgen degradation in aerobic sewage. Sewage samples collected from the Dihua Sewage Treatment Plant (Taipei, Taiwan) were aerobically incubated with testosterone (1 mM). Androgen metabolite analysis revealed that bacteria adopt the 9, 10-seco pathway to degrade testosterone. A metagenomic analysis indicated the apparent enrichment of Comamonas spp. (mainly C. testosteroni) and Pseudomonas spp. in sewage incubated with testosterone. We used the degenerate primers derived from the meta-cleavage dioxygenase gene (tesB) of various proteobacteria to track this essential catabolic gene in the sewage. The amplified sequences showed the highest similarity (87–96%) to tesB of C. testosteroni. Using quantitative PCR, we detected a remarkable increase of the 16S rRNA and catabolic genes of C. testosteroni in the testosterone-treated sewage. Together, our data suggest that C. testosteroni, the model microorganism for aerobic testosterone degradation, plays a role in androgen biodegradation in aerobic sewage. PMID:27734937

  1. The role of hydrogen for Sulfurimonas denitrificans' metabolism.

    PubMed

    Han, Yuchen; Perner, Mirjam

    2014-01-01

    Sulfurimonas denitrificans was originally isolated from coastal marine sediments. It can grow with thiosulfate and nitrate or sulfide and oxygen. Recently sequencing of its genome revealed that it encodes periplasmic and cytoplasmic [NiFe]-hydrogenases but the role of hydrogen for its metabolism has remained unknown. We show the first experimental evidence that S. denitrificans can indeed express a functional hydrogen uptake active hydrogenase and can grow on hydrogen. In fact, under the provided conditions it grew faster and denser on hydrogen than on thiosulfate alone and even grew with hydrogen in the absence of reduced sulfur compounds. In our experiments, at the time points tested, the hydrogen uptake activity appeared to be related to the periplasmic hydrogenase and not to the cytoplasmic hydrogenase. Our data suggest that under the provided conditions S. denitrificans can grow more efficiently with hydrogen than with thiosulfate. PMID:25170905

  2. Oxidation of hydrogen sulfide by continuous cultures of Thiobacillus denitrificans

    SciTech Connect

    Sublette, K.L.; Sylvester, N.D.

    1987-04-01

    In order to study the anaerobic oxidation of H/sub 2/S by T. denitrificans under constant growth conditions the organism was cultured in a continuous stirred tank reactor. A series of experiments was conducted having three purposes: 1) to identify any unforeseen problems which might be associated with growth of T. denitrificans on a continuous basis; 2) to examine the effect of metabolic state and environment on the maximum loading of H/sub 2/S; and 3) to examine the relationship between growth rate and yield of biomass per mole H/sub 2/S oxidized.

  3. Halobacterium denitrificans sp. nov. - An extremely halophilic denitrifying bacterium

    NASA Technical Reports Server (NTRS)

    Tomlinson, G. A.; Jahnke, L. L.; Hochstein, L. I.

    1986-01-01

    Halobacterium denitrificans was one of several carbohydrate-utilizing, denitrifying, extremely halophilic bacteria isolated by anaerobic enrichment in the presence of nitrate. Anaerobic growth took place only when nitrate (or nitrite) was present and was accompanied by the production of dinitrogen. In the presence of high concentrations of nitrate (i.e., 0.5 percent), nitrous oxide and nitrite were also detected. When grown aerobically in a mineral-salts medium containing 0.005 percent yeast extract, H. denitrificans utilized a variety of carbohydrates as sources of carbon and energy. In every case, carbohydrate utilization was accompanied by acid production.

  4. Halobacterium denitrificans sp. nov., an extremely halophilic denitrifying bacterium

    NASA Technical Reports Server (NTRS)

    Tomlinson, G. A.; Jahnke, L. L.; Hochstein, L. I.

    1986-01-01

    Halobacterium denitrificans was one of several carbohydrate-utilizing, denitrifying, extremely halophilic bacteria isolated by anaerobic enrichment in the presence of nitrate. Anaerobic growth took place only when nitrate (or nitrite) was present and was accompanied by the production of dinitrogen. In the presence of high concentrations of nitrate (i.e., 0.5 percent), nitrous oxide and nitrite were also detected. When grown aerobically in a mineral-salts medium containing 0.005 percent yeast extract, H. denitrificans utilized a variety of carbohydrates as sources of carbon and energy. In every case, carbohydrate utilization was accompanied by acid production.

  5. Novel cyanide-hydrolyzing enzyme from Alcaligenes xylosoxidans subsp. denitrificans.

    PubMed Central

    Ingvorsen, K; Højer-Pedersen, B; Godtfredsen, S E

    1991-01-01

    A cyanide-metabolizing bacterium, strain DF3, isolated from soil was identified as Alcaligenes xylosoxidans subsp. denitrificans. Whole cells and cell extracts of strain DF3 catalyzed hydrolysis of cyanide to formate and ammonia (HCN + 2H2O----HCOOH + NH3) without forming formamide as a free intermediate. The cyanide-hydrolyzing activity was inducibly produced in cells during growth in cyanide-containing media. Cyanate (OCN-) and a wide range of aliphatic and aromatic nitriles were not hydrolyzed by intact cells of A. xylosoxidans subsp. denitrificans DF3. Strain DF3 hydrolyzed cyanide with great efficacy. Thus, by using resting induced cells at a concentration of 11.3 mg (dry weight) per ml, the cyanide concentration could be reduced from 0.97 M (approximately 25,220 ppm) to less than 77 nM (approximately 0.002 ppm) in 55 h. Enzyme purification established that cyanide hydrolysis by A. xylosoxidans subsp. denitrificans DF3 was due to a single intracellular enzyme. The soluble enzyme was purified approximately 160-fold, and the first 25 NH2-terminal amino acids were determined by automated Edman degradation. The molecular mass of the active enzyme (purity, greater than 97% as determined by amino acid sequencing) was estimated to be greater than 300,000 Da. The cyanide-hydrolyzing enzyme of A. xylosoxidans subsp. denitrificans DF3 was tentatively named cyanidase to distinguish it from known nitrilases (EC 3.5.5.1) which act on organic nitriles. Images PMID:1872607

  6. Analyses of a Polyhydroxyalkanoic Acid Granule-Associated 16-Kilodalton Protein and Its Putative Regulator in the pha Locus of Paracoccus denitrificans

    PubMed Central

    Maehara, Akira; Ueda, Shunsaku; Nakano, Hideo; Yamane, Tsuneo

    1999-01-01

    The polyhydroxyalkanoic acid (PHA) granule-associated 16-kDa protein (GA16 protein) of Paracoccus denitrificans was identified, and its corresponding gene was cloned and analyzed at the molecular level. The N-terminal amino acid sequence of GA16 protein revealed that its structural gene is located downstream from the PHA synthase gene (phaCPd) cloned recently (S. Ueda, T. Yabutani, A. Maehara, and T. Yamane, J. Bacteriol. 178:774–779, 1996). Gene walking around phaCPd revealed two new open reading frames (ORFs) possibly related to PHA synthesis, one of which was the phaPPd gene, encoding GA16 protein, and the other was the phaRPd gene, encoding a protein that is putatively involved in the regulation of the expression of phaPPd. Overproduction of PhaPPd was observed in Escherichia coli carrying phaPPd, but the overproduction was not observed in the presence of phaRPd. Coexpression of phaPPd and PHA biosynthesis genes in E. coli caused increases in both the number of poly-(3-hydroxybutyric acid) (PHB) granules and PHB content and caused decreases in both the size of the granules and the molecular weight of PHB. GA16 protein was considered a phasin protein. The phaRPd gene had significant similarities to stdC, a possible transcriptional factor of Comamonas testosteroni, as well as to other ORFs of unknown function previously found in other PHA-synthetic bacteria. PMID:10217786

  7. Atypical Polyphosphate Accumulation by the Denitrifying Bacterium Paracoccus denitrificans

    PubMed Central

    Barak, Yoram; van Rijn, Jaap

    2000-01-01

    Polyphosphate accumulation by Paracoccus denitrificans was examined under aerobic, anoxic, and anaerobic conditions. Polyphosphate synthesis by this denitrifier took place with either oxygen or nitrate as the electron acceptor and in the presence of an external carbon source. Cells were capable of poly-β-hydroxybutyrate (PHB) synthesis, but no polyphosphate was produced when PHB-rich cells were incubated under anoxic conditions in the absence of an external carbon source. By comparison of these findings to those with polyphosphate-accumulating organisms thought to be responsible for phosphate removal in activated sludge systems, it is concluded that P. denitrificans is capable of combined phosphate and nitrate removal without the need for alternating anaerobic/aerobic or anaerobic/anoxic switches. Studies on additional denitrifying isolates from a denitrifying fluidized bed reactor suggested that polyphosphate accumulation is widespread among denitrifiers. PMID:10698794

  8. Isolation and algicidal characterization of Bowmanella denitrificans S088 against Chlorella vulgaris.

    PubMed

    Jiang, Xiao; Ren, Chunhua; Hu, Chaoqun; Zhao, Zhe

    2014-02-01

    One strain of algicidal bacterium, named as S088, was isolated from the intestine of healthy sea cucumbers (Stichopus horrens) in the South China Sea. Based on the analysis of its biochemical characteristics and 16S rDNA gene sequence, S088 was identified as Bowmanella denitrificans. Importantly, the algicidal activity of S088 on Chlorella vulgaris was characterized in this study. The initial densities of bacterial and algal cell showed strong influence on the removal rates of chlorophyll a. When the strain S088 was cultured under a complete darkness condition at 30 °C, its algicidal activity reached the highest level. Furthermore, it was found that the filtered supernatant from bacterial cultures had full algicidal activity, suggesting that the secreted compounds from S088 are involved in the observed algicidal action of S088. Moreover, the algicidal compounds were heat tolerant and had no cytotoxicity against fish cells, indicating that S088 would have a promising application as a safe probiotics for S. horrens. Finally, this is the first report about the algicidal activities in B. denitrificans.

  9. Aerobic and anaerobic growth of Paracoccus denitrificans on methanol.

    PubMed

    Bamforth, C W; Quayle, J R

    1978-10-01

    1. The dye-linked methanol dehydrogenase from Paracoccus denitrificans grown aerobically on methanol has been purified and its properties compared with similar enzymes from other bacteria. It was shown to be specific and to have high affinity for primary alcohols and formaldehyde as substrate, ammonia was the best activator and the enzyme could be linked to reduction of phenazine methosulphate. 2. Paracoccus denitrificans could be grown anaerobically on methanol, using nitrate or nitrite as electron acceptor. The methanol dehydrogenase synthesized under these conditions could not be differentiated from the aerobically-synthesized enzyme. 3. Activities of methanol dehydrogenase, formaldehyde dehydrogenase, formate dehydrogenase, nitrate reductase and nitrite reductase were measured under aerobic and anaerobic growth conditions. 4. Difference spectra of reduced and oxidized cytochromes in membrane and supernatant fractions of methanol-grown P. denitrificans were measured. 5. From the results of the spectral and enzymatic analyses it has been suggested that anaerobic growth on methanol/nitrate is made possible by reduction of nitrate to nitrite using electrons derived from the pyridine nucleotide-linked dehydrogenations of formaldehyde and formate, the nitrite so produced then functioning as electron acceptor for methanol dehydrogenase via cytochrome c and nitrite reductase. PMID:718372

  10. Comamonas testosteroni-associated peritonitis in a pediatric peritoneal dialysis patient.

    PubMed

    Parolin, Mattia; Baraldi, Maura; Valentini, Elena; Murer, Luisa; Vidal, Enrico

    2016-03-01

    Comamonas testosteroni (C. testosteroni) has been rarely observed as an infectious agent in clinical practice. Few reports described its potential pathogenicity in bloodstream and abdominal infections. Here, we report our experience in the treatment of a C. testosteroni-associated peritonitis in a four-year-old girl receiving chronic peritoneal dialysis (PD). The organism was shown to be highly susceptible to appropriate antibiotic therapy. Infection responded promptly and the patient was managed conservatively without withdrawal from PD.

  11. Paracoccus denitrificans possesses two BioR homologs having a role in regulation of biotin metabolism

    PubMed Central

    Feng, Youjun; Kumar, Ritesh; Ravcheev, Dmitry A; Zhang, Huimin

    2015-01-01

    Recently, we determined that BioR, the GntR family of transcription factor, acts as a repressor for biotin metabolism exclusively distributed in certain species of α-proteobacteria, including the zoonotic agent Brucella melitensis and the plant pathogen Agrobacterium tumefaciens. However, the scenario is unusual in Paracoccus denitrificans, another closely related member of the same phylum α-proteobacteria featuring with denitrification. Not only does it encode two BioR homologs Pden_1431 and Pden_2922 (designated as BioR1 and BioR2, respectively), but also has six predictive BioR-recognizable sites (the two bioR homolog each has one site, whereas the two bio operons (bioBFDAGC and bioYB) each contains two tandem BioR boxes). It raised the possibility that unexpected complexity is present in BioR-mediated biotin regulation. Here we report that this is the case. The identity of the purified BioR proteins (BioR1 and BioR2) was confirmed with LC-QToF-MS. Phylogenetic analyses combined with GC percentage raised a possibility that the bioR2 gene might be acquired by horizontal gene transfer. Gel shift assays revealed that the predicted BioR-binding sites are functional for the two BioR homologs, in much similarity to the scenario seen with the BioR site of A. tumefaciens bioBFDAZ. Using the A. tumefaciens reporter system carrying a plasmid-borne LacZ fusion, we revealed that the two homologs of P. denitrificans BioR are functional repressors for biotin metabolism. As anticipated, not only does the addition of exogenous biotin stimulate efficiently the expression of bioYB operon encoding biotin transport/uptake system BioY, but also inhibits the transcription of the bioBFDAGC operon resembling the de novo biotin synthetic pathway. EMSA-based screening failed to demonstrate that the biotin-related metabolite is involved in BioR-DNA interplay, which is consistent with our former observation with Brucella BioR. Our finding defined a complex regulatory network for biotin

  12. Comamonas testosteroni uses a chemoreceptor for tricarboxylic acid cycle intermediates to trigger chemotactic responses towards aromatic compounds.

    PubMed

    Ni, Bin; Huang, Zhou; Fan, Zheng; Jiang, Cheng-Ying; Liu, Shuang-Jiang

    2013-11-01

    Bacterial chemotaxis towards aromatic compounds has been frequently observed; however, knowledge of how bacteria sense aromatic compounds is limited. Comamonas testosteroni CNB-1 is able to grow on a range of aromatic compounds. This study investigated the chemotactic responses of CNB-1 to 10 aromatic compounds. We constructed a chemoreceptor-free, non-chemotactic mutant, CNB-1Δ20, by disruption of all 19 putative methyl-accepting chemotaxis proteins (MCPs) and the atypical chemoreceptor in strain CNB-1. Individual complementation revealed that a putative MCP (tagged MCP2201) was involved in triggering chemotaxis towards all 10 aromatic compounds. The recombinant sensory domain of MCP2201 did not bind to 3- or 4-hydroxybenzoate, protocatechuate, catechol, benzoate, vanillate and gentisate, but bound oxaloacetate, citrate, cis-aconitate, isocitrate, α-ketoglutarate, succinate, fumarate and malate. The mutant CNB-1ΔpmdF that lost the ability to metabolize 4-hydroxybenzoate and protocatechuate also lost its chemotactic response to these compounds, suggesting that taxis towards aromatic compounds is metabolism-dependent. Based on the ligand profile, we proposed that MCP2201 triggers taxis towards aromatic compounds by sensing TCA cycle intermediates. Our hypothesis was further supported by the finding that introduction of the previously characterized pseudomonad chemoreceptor (McpS) for TCA cycle intermediates into CNB-1Δ20 likewise triggered chemotaxis towards aromatic compounds.

  13. Substrate uptake and subcellular compartmentation of anoxic cholesterol catabolism in Sterolibacterium denitrificans.

    PubMed

    Lin, Ching-Wen; Wang, Po-Hsiang; Ismail, Wael; Tsai, Yu-Wen; El Nayal, Ashraf; Yang, Chia-Ying; Yang, Fu-Chun; Wang, Chia-Hsiang; Chiang, Yin-Ru

    2015-01-01

    Cholesterol catabolism by actinobacteria has been extensively studied. In contrast, the uptake and catabolism of cholesterol by Gram-negative species are poorly understood. Here, we investigated microbial cholesterol catabolism at the subcellular level. (13)C metabolomic analysis revealed that anaerobically grown Sterolibacterium denitrificans, a β-proteobacterium, adopts an oxygenase-independent pathway to degrade cholesterol. S. denitrificans cells did not produce biosurfactants upon growth on cholesterol and exhibited high cell surface hydrophobicity. Moreover, S. denitrificans did not produce extracellular catabolic enzymes to transform cholesterol. Accordingly, S. denitrificans accessed cholesterol by direction adhesion. Cholesterol is imported through the outer membrane via a putative FadL-like transport system, which is induced by neutral sterols. The outer membrane steroid transporter is able to selectively import various C27 sterols into the periplasm. S. denitrificans spheroplasts exhibited a significantly higher efficiency in cholest-4-en-3-one-26-oic acid uptake than in cholesterol uptake. We separated S. denitrificans proteins into four fractions, namely the outer membrane, periplasm, inner membrane, and cytoplasm, and we observed the individual catabolic reactions within them. Our data indicated that, in the periplasm, various periplasmic and peripheral membrane enzymes transform cholesterol into cholest-4-en-3-one-26-oic acid. The C27 acidic steroid is then transported into the cytoplasm, in which side-chain degradation and the subsequent sterane cleavage occur. This study sheds light into microbial cholesterol metabolism under anoxic conditions.

  14. Substrate Uptake and Subcellular Compartmentation of Anoxic Cholesterol Catabolism in Sterolibacterium denitrificans*

    PubMed Central

    Lin, Ching-Wen; Wang, Po-Hsiang; Ismail, Wael; Tsai, Yu-Wen; El Nayal, Ashraf; Yang, Chia-Ying; Yang, Fu-Chun; Wang, Chia-Hsiang; Chiang, Yin-Ru

    2015-01-01

    Cholesterol catabolism by actinobacteria has been extensively studied. In contrast, the uptake and catabolism of cholesterol by Gram-negative species are poorly understood. Here, we investigated microbial cholesterol catabolism at the subcellular level. 13C metabolomic analysis revealed that anaerobically grown Sterolibacterium denitrificans, a β-proteobacterium, adopts an oxygenase-independent pathway to degrade cholesterol. S. denitrificans cells did not produce biosurfactants upon growth on cholesterol and exhibited high cell surface hydrophobicity. Moreover, S. denitrificans did not produce extracellular catabolic enzymes to transform cholesterol. Accordingly, S. denitrificans accessed cholesterol by direction adhesion. Cholesterol is imported through the outer membrane via a putative FadL-like transport system, which is induced by neutral sterols. The outer membrane steroid transporter is able to selectively import various C27 sterols into the periplasm. S. denitrificans spheroplasts exhibited a significantly higher efficiency in cholest-4-en-3-one-26-oic acid uptake than in cholesterol uptake. We separated S. denitrificans proteins into four fractions, namely the outer membrane, periplasm, inner membrane, and cytoplasm, and we observed the individual catabolic reactions within them. Our data indicated that, in the periplasm, various periplasmic and peripheral membrane enzymes transform cholesterol into cholest-4-en-3-one-26-oic acid. The C27 acidic steroid is then transported into the cytoplasm, in which side-chain degradation and the subsequent sterane cleavage occur. This study sheds light into microbial cholesterol metabolism under anoxic conditions. PMID:25418128

  15. Optimization of intracellular product release from Neisseria denitrificans using microfluidizer.

    PubMed

    Stupak, Robert; Makauskas, Nerijus; Radzevičius, Kostas; Valančius, Zenonas

    2015-01-01

    Disruption of Neisseria denitrificans cells by microfluidizer was optimized using a factorial experiments design. The pH, pretreatment time, cell concentration, NaCl, ethylenediamine tetraacetic acid (EDTA) and Triton X-100 concentrations showed significant impact on disruption process and the process was optimized using central composite design and response surface methodology (RSM). Investigation revealed optimum conditions: 90 min pretreatment at pH 9.0 containing 110 g L(-1) cells (dry cell weight), 50 mM NaCl, 10 mM EDTA, and 0.2% Triton X-100. At optimized conditions, the disruption rate increased twofold, up to 5.62 ± 0.27 × 10(-3) MPa(-a); meanwhile, yield of intracellular content was increased by 26%, with 1 g of cells resulting in 113.2 ± 8.2 mg proteins, 12.1 ± 0.7 mg nucleic acids, 21.0 ± 1.2 mg polysaccharides, 0.99 ± 0.08 kU glucose-6-phosphate dehydrogenase (G6PD), and 10,100 ± 110 kU restriction endonuclease NdeI endonuclease. Particle size distribution analysis revealed nearly twofold larger cell lysate particles with diameter of 120 nm. For optimal release of intracellular content, 9200 J/g of energy was needed (95% confidence), yielding 6900 J/g energy savings. Model equations generated from RSM on cell disruption of N. denitrificans were found adequate to determine significant factors and its interaction. The results showed that optimized combination of known pretreatment and disruption methods could considerably improve cell disruption efficiency.

  16. Draft Genome Sequence of Comamonas thiooxydans Strain PHE2-6 (NBRC 110656), a Chlorinated-Ethene-Degrading Bacterium

    PubMed Central

    Shimodaira, Jun; Yonezuka, Kenta; Tabata, Michiro; Nagase, Shun; Kasai, Daisuke; Hosoyama, Akira; Yamazoe, Atsushi; Fujita, Nobuyuki

    2016-01-01

    Comamonas thiooxydans strain PHE2-6 (NBRC 110656), which was isolated from a trichloroethene-contaminated site in Japan, utilizes phenol as a sole source of carbon and cometabolizes cis- and trans-dichloroethenes. We report here the draft genome sequence of this strain, containing 5,309,680 bp, with 60.6% G+C content. PMID:27340052

  17. Production of R-(-)-Ketoprofen from an Amide Compound by Comamonas acidovorans KPO-2771-4

    PubMed Central

    Yamamoto, K.; Otsubo, K.; Matsuo, A.; Hayashi, T.; Fujimatsu, I.; Komatsu, K.

    1996-01-01

    R-(-)-2-(3(prm1)-Benzoylphenyl)propionic acid [R-(-)-ketoprofen] was produced from racemic 2-(3(prm1)-benzoylphenyl)propionamide (keto-amide) by the isolated bacterial strain Comamonas acidovorans KPO-2771-4. Sodium fumarate as the carbon source and 2-azacyclononanone or isobutyronitrile as the enhancer in the culture medium were effective for bacterial growth and the enhancement of R-(-)-ketoprofen-producing activity. R-(-)-Ketoprofen produced from the keto-amide by resting cells was present in 99% enantiomeric exess. C. acidovorans KPO-2771-4 has an R-enantioselective amidase for keto-amide because the purified amidase from the bacterium hydrolyzed keto-amide, producing optically pure R-ketoprofen and ammonia. PMID:16535206

  18. Production of 3-hydroxypropionic acid from glycerol by recombinant Pseudomonas denitrificans.

    PubMed

    Zhou, Shengfang; Catherine, Christy; Rathnasingh, Chelladurai; Somasundar, Ashok; Park, Sunghoon

    2013-12-01

    3-Hydroxypropionic acid (3-HP) can be produced from glycerol through two sequential enzymatic reactions that are catalyzed by a coenzyme B12 -dependent glycerol dehydratase and an NAD(P)(+) -dependent aldehyde dehydrogenase (ALDH), respectively. Pseudomonas denitrificans synthesizes coenzyme B12 under aerobic conditions, where NAD(P)(+) is regenerated efficiently. Hence, it is considered an ideal host for the production of 3-HP from glycerol under aerobic conditions. In this study, recombinant strains of P. denitrificans were developed and their potential for the production of 3-HP from glycerol was evaluated. When the enzymes, glycerol dehydratase (DhaB) and glycerol dehydratase reactivase (GdrAB), of Klebsiella pneumoniae were expressed heterologously, P. denitrificans could produce 3-HP at 37.7 mmol/L with 62% (mol/mol) yield on glycerol. Glucose was required as the carbon and energy sources for cell growth. The overexpression of heterologous ALDH was not essential; however, the titer and yield of 3-HP were improved to 54.7 mmol/L and 67% (mol/mol), respectively, when an ALDH gene (puuC) from K. pneumoniae was overexpressed. One serious drawback hindering the use of P. denitrificans as a recombinant host for 3-HP production is that it oxidizes 3-HP to malonate and utilizes 3-HP as a carbon source for growth. This is the first report on the development and use of recombinant P. denitrificans for 3-HP production from glycerol.

  19. Improved vitamin B12 fermentation process by adding rotenone to regulate the metabolism of Pseudomonas denitrificans.

    PubMed

    Cheng, Xin; Chen, Wei; Peng, Wei-Fu; Li, Kun-Tai

    2014-06-01

    Our previous research had revealed that the dissolved oxygen limitation was more favorable for vitamin B12 fermentation, due to its inducement to the increased glycolytic flux in Pseudomonas denitrificans. In this paper, a novel strategy was implemented to further investigate the metabolic characteristics of P. denitrificans under different oxygen supply levels, by exogenously adding rotenone (a respiratory chain inhibitor interfering with the oxygen consumption) to the fermentation broths. Compared to the fermentation process without rotenone treatment, it was observed that 5 mg/L rotenone treatment could significantly strengthen the glycolytic flux of P. denitrificans via activating the key glycolytic enzymes (phosphofructokinase and pyruvate kinase), resulting in the accelerated generations of anterior precursors (glutamate and 5-aminolevulinic acid) for vitamin B12 biosynthesis. Although 5 mg/L rotenone treatment had a negative effect on cell growth of P. denitrificans, the vitamin B12 yield was increased from 48.28 ± 0.62 mg/L to 54.70 ± 0.45 mg/L, which further proved that an increased glycolytic flux in P. denitrificans was a consequence of higher vitamin B12 production. PMID:24687557

  20. Comamonas sp. halotolerant bacterium from industrial zone of Jovein of Sabzevar introduced as good candidate to remove industrial pollution

    PubMed Central

    Ghanbarinia, Fahimeh; Kheirbadi, Mitra; Mollania, Nasrin

    2015-01-01

    Background and Objectives: Heavy metals are considered as high risk biocides due to their harmful effects on human health, the environment and other living organisms. Bacterial strains showing resistance to heavy metals has been used for removing such toxic materials from the environment. In this study we isolated and characterized a heavy metals-resistance halophilic bacterial strains from Kal shoor Jovein of Sabzevar, one of the industrial zone of Khorasan-e-Razavi province in Iran and has naturally saline oils. Materials and Methods: Strain JC-66 is heavy metals-resistance halophilic bacterial strains isolated from Kal shoor Jovein of Sabzevar. The 16S rDNA gene was sequenced to identify this bacterium. The appropriate conditions for its potency to remove the lead were tested in various temprature, pH and agitation speed. The resistance mechanism of JC-66 to lead were investigated. Results: JC-66 is a Comamonas sp. according to 16S rDNA sequence analysis. Based on minimum inhibitory concentration (MIC) results, the isolated strain has high resistance to the lead metal. The optimal condition for lead removal was exhibited in neutral medium (pH 7) incubation temperature 37 °C, and shaking rate of 180 rpm for JC-66. X-Ray Diffraction results also are indicative of adsorption mechanism to lead metal uptake. Plasmid extraction was performed to confirm the role of plasmids in bacterial resistance to lead. Conclusion: It can be concluded that the mechanism of resistance to heavy metals in the studied strain, is the result of an expression plasmid, and adsorption. It was concluded that JC-66 is able to be one of the best candidates to remove industrial pollution because it showed high resistance to lead. PMID:26719784

  1. Paracoccus denitrificans for the effluent recycling during continuous denitrification of liquid food.

    PubMed

    Tippkötter, Nils; Roikaew, Wipa; Ulber, Roland; Hoffmann, Alexander; Denzler, Hans-Jörg; Buchholz, Heinrich

    2010-01-01

    Nitrate is an undesirable component of several foods. A typical case of contamination with high nitrate contents is whey concentrate, containing nitrate in concentrations up to 25 l. The microbiological removal of nitrate by Paracoccus denitrificans under formation of harmless nitrogen in combination with a cell retention reactor is described here. Focus lies on the resource-conserving design of a microbal denitrification process. Two methods are compared. The application of polyvinyl alcohol-immobilized cells, which can be applied several times in whey feed, is compared with the implementation of a two step denitrification system. First, the whey concentrate's nitrate is removed by ion exchange and subsequently the eluent regenerated by microorganisms under their retention by crossflow filtration. Nitrite and nitrate concentrations were determined by reflectometric color measurement with a commercially available Reflectoquant device. Correction factors for these media had to be determined. During the pilot development, bioreactors from 4 to 250 mg x L(-1) and crossflow units with membrane areas from 0.02 to 0.80 m(2) were examined. Based on the results of the pilot plants, a scaling for the exemplary process of denitrifying 1,000 tons per day is discussed. PMID:20187124

  2. Identification of a blue copper protein from Hyphomicrobium denitrificans and its functions in the periplasm.

    PubMed

    Hira, Daisuke; Nojiri, Masaki; Yamaguchi, Kazuya; Suzuki, Shinnichiro

    2007-09-01

    It has been known that the methylotrophic denitrifying bacteria have the specific electron transfer chains, involving in 'methanol oxidation' and 'denitrification', in the periplasm. Recently, a unique blue copper protein (HdBCP) has been isolated from the methanol-grown methylotrophic denitrifying bacterium, Hyphomicrobium denitrificans. HdBCP is a 14.5 kDa protein and contains one copper atom in the molecule. The electronic absorption spectrum of HdBCP exhibits two absorption maxima near 450 and 750 nm comparable with the intense 600 nm band (epsilon(450)/epsilon(600) = ca. 0.9). The rhombic electron paramagnetic resonance spectrum shows clearly that the copper centre is a 'perturbed' type 1 copper geometry. Stopped-flow kinetics indicates that HdBCP accepts efficiently an electron from cytochrome c(L) (k(2) = 4.0 x 10(6) M(-1) s(-1) at 25.0 degrees C), which is a physiological electron acceptor for methanol dehydrogenase. According to cloning and DNA sequencing of the structural gene, the deduced amino acid sequence shows significant similarities with pseudoazurins, which are a physiological electron donor for Cu-containing nitrite reductase from the denitrifying bacteria. Based on these results, we discuss the role of HdBCP in the electron-flow system, which link 'methanol oxidation' and 'denitrification' together. PMID:17646178

  3. Biodegradation of Benzene, Toluene, Ethylbenzene, and o-, m-, and p-Xylenes by the Newly Isolated Bacterium Comamonas sp. JB.

    PubMed

    Jiang, Bei; Zhou, Zunchun; Dong, Ying; Tao, Wei; Wang, Bai; Jiang, Jingwei; Guan, Xiaoyan

    2015-07-01

    A bacterium designated strain JB, able to degrade six benzene, toluene, ethylbenzene, and o-, m-, and p-xylene (BTEX) compounds, was isolated from petroleum-contaminated soil. Taxonomic analyses showed that the isolate belonged to Comamonas, and until now, the genus Comamonas has not included any known BTEX degraders. The BTEX biodegradation rate was slightly low on the mineral salt medium (MSM), but adding a small amount of yeast extract greatly enhanced the biodegradation. The relationship between specific degradation rate and individual BTEX was described well by Michaelis-Menten kinetics. The treatment of petrochemical wastewater containing BTEX mixture and phenol was shown to be highly efficient by BTEX-grown JB. In addition, toxicity assessment indicated the treatment of the petrochemical wastewater by BTEX-grown JB led to less toxicity than untreated wastewater.

  4. Regulation of oxidative phosphorylation: the flexible respiratory network of Paracoccus denitrificans.

    PubMed

    Van Spanning, R J; de Boer, A P; Reijnders, W N; De Gier, J W; Delorme, C O; Stouthamer, A H; Westerhoff, H V; Harms, N; van der Oost, J

    1995-10-01

    Paracoccus denitrificans is a facultative anaerobic bacterium that has the capacity to adjust its metabolic infrastructure, quantitatively and/or qualitatively, to the prevailing growth condition. In this bacterium the relative activity of distinct catabolic pathways is subject to a hierarchical control. In the presence of oxygen the aerobic respiration, the most efficient way of electron transfer-linked phosphorylation, has priority. At high oxygen tensions P. denitrificans synthesizes an oxidase with a relatively low affinity for oxygen, whereas under oxygen limitation a high-affinity oxidase appears specifically induced. During anaerobiosis, the pathways with lower free energy-transducing efficiency are induced. In the presence of nitrate, the expression of a number of dehydrogenases ensures the continuation of oxidative phosphorylation via denitrification. After identification of the structural components that are involved in both the aerobic and the anaerobic respiratory networks of P. denitrificans, the intriguing next challenge is to get insight in its regulation. Two transcription regulators have recently been demonstrated to be involved in the expression of a number of aerobic and/or anaerobic respiratory complexes in P. denitrificans. Understanding of the regulation machinery is beginning to emerge and promises much excitement in discovery. PMID:8718455

  5. Mode of action of copper complexes of some 2,2'-bipyridyl analogs on Paracoccus denitrificans.

    PubMed Central

    Smit, H; van der Goot, H; Nauta, W T; Pijper, P J; Balt, S; de Bolster, M W; Stouthamer, A H; Verheul, H; Vis, R D

    1980-01-01

    Copper complexes of 2,2'-bipyridyl and related compounds and CuSO4 inhibited the growth of paracoccus denitrificans. The copper(I) complex of 2,9-dimethyl-1,10-phenanthroline [Cu(DMP)2NO3] showed the highest activity, whereas the copper(II) complex of 1,10-phenanthroline and CuSO4 inhibited the growth to a lesser extent. The uncomplexed ligands (1,10-phenanthroline and 2,9-dimethyl-1,10-phenanthroline) showed little activity, but in the presence of noninhibitory amounts of CuSO4 this activity increased markedly. Copper ions therefore proved to be essential for the growth-inhibitor effect. The extent of inhibition appeared to be strongly dependent on the initial cell density and on the growth medium. No selective inhibition of deoxyribonucleic acid, ribonucleic acid, or protein synthesis was observed with Cu(DMP)2NO3. Respiratory electron transport of P. denitrificans appeared to be strongly inhibited by Cu(DMP)2NO3 and to a somewhat lesser extent by CuSO4. Both aerobic and anaerobic respirations were inhibited to the same extent, and from the cytochrome redox kinetics it is concluded that the site of this inhibition in the respiratory electron transport chain must be located before cytochrome b. Cu(DMP)2NO3 did not significantly influence the H+/O ratio with whole cells of P. denitrificans, suggesting that the efficiency of oxidative phosphorylation is not affected by CU(DMP)2NO3. Growing cultures of P. denitrificans showed a decrease in intracellular potassium ion content in the presence of increasing amounts of Cu(DMP)2NO3. It is concluded that interference with the cytoplasmic membrane, resulting in inhibition of respiratory electron transport, probably constitutes the main mode of action of copper complexes of 2,2'-bipyridyl analogs on P. denitrificans. PMID:6778379

  6. Kinetics of iron acquisition from ferric siderophores by Paracoccus denitrificans.

    PubMed Central

    Bergeron, R J; Weimar, W R

    1990-01-01

    The kinetics of iron accumulation by iron-starved Paracoccus denitrificans during the first 2 min of exposure to 55Fe-labeled ferric siderophore chelates is described. Iron is acquired from the ferric chelate of the natural siderophore L-parabactin in a process exhibiting biphastic kinetics by Lineweaver-Burk analysis. The kinetic data for 1 microM less than [Fe L-parabactin] less than 10 microM fit a regression line which suggests a low-affinity system (Km = 3.9 +/- 1.2 microM, Vmax = 494 pg-atoms of 55Fe min-1 mg of protein-1), whereas the data for 0.1 microM less than or equal to [Fe L-parabactin] less than or equal to 1 microM fit another line consistent with a high-affinity system (Km = 0.24 +/- 0.06 microM, Vmax = 108 pg-atoms of 55Fe min-1 mg of protein-1). The Km of the high-affinity uptake is comparable to the binding affinity we had previously reported for the purified ferric L-parabactin receptor protein in the outer membrane. In marked contrast, ferric D-parabactin data fit a single regression line corresponding to a simple Michaelis-Menten process with comparatively low affinity (Km = 3.1 +/- 0.9 microM, Vmax = 125 pg-atoms of 55Fe min-1 mg of protein-1). Other catecholamide siderophores with an intact oxazoline ring derived from L-threonine (L-homoparabactin, L-agrobactin, and L-vibriobactin) also exhibit biphasic kinetics with a high-affinity component similar to ferric L-parabactin. Circular dichroism confirmed that these ferric chelates, like ferric L-parabactin, exist as the lambda enantiomers. The A forms ferric parabactin (ferrin D- and L-parabactin A), in which the oxazoline ring is hydrolyzed to the open-chain threonyl structure, exhibit linear kinetics with a comparatively high Km (1.4 +/- 0.3 microM) and high Vmax (324 pg-atoms of 55Fe min-1 of protein-1). Furthermore, the marked stereospecificity seen between ferric D- and L-parabactins is absent; i.e., iron acquisition from ferric parabactin A is non stereospecific. The mechanistic

  7. Colonization of an acid resistant Kingella denitrificans in the stomach may contribute to gastric dysbiosis by Helicobacter pylori.

    PubMed

    Okamoto, Takeshi; Hayashi, Yasuhiro; Mizuno, Hidekazu; Yanai, Hideo; Nishikawa, Jun; Nakazawa, Teruko; Iizasa, Hisashi; Jinushi, Masahisa; Sakaida, Isao; Yoshiyama, Hironori

    2014-03-01

    In the stomach of a gastric ulcer patient who had been administered an anti-acid, a gram-negative and urease-negative bacillus similar in size to Helicobacter pylori was infected together with H. pylori. According to biochemical test and 16S rRNA gene analysis, the urease-negative bacterium was identified as Kingella denitrificans, a human nasopharyngeal commensal. In contrast to the standard strain of K. denitrificans, the isolate showed catalase activity, did not produce acid from glucose, and exhibited acid tolerance. Acid tolerance of H. pylori was increased by cocultivation with the K. denitrificans isolate, but not with other isolates of K. denitrificans. Disruption of physiological and immunological niche by dysbiotic colonization of bacterium may provide pathological attributes to human stomach. Collectively, a careful administration of anti-acids to the elderly, especially those with atrophic gastritis, is necessary to avoid repression of the gastric barrier to bacteria.

  8. Continuous degradation of dimethyl sulfoxide to sulfate ion by Hyphomicrobium denitrificans WU-K217.

    PubMed

    Murakami-Nitta, Takako; Kurimura, Hiroyuki; Kirimura, Kohtaro; Kino, Kuniki; Usami, Shoji

    2002-01-01

    With the objective of removing dimethyl sulfoxide (DMSO) contained in wastewater from semiconductor or liquid crystal display factories, biodegradation of DMSO, particularly at a low concentration, was examined. Through the screening of DMSO-degrading microorganisms, Hyphomicrobium denitrificans WU-K217 utilizing DMSO as the sole source of carbon was isolated from soil. DMSO at less than 20 mM was degraded to sulfate ion by WU-K217 with 100% molar conversion ratio based on DMSO added during 60-h cultivation at 30 degrees C under aerobic conditions. Even in the presence of 116 mM or 225 mM DMSO, WU-K217 showed growth although the amount of DMSO degraded was only 33 mM or 10 mM, respectively. Similar to the growing cells, the resting cells of WU-K217 degraded DMSO at over a wide range of temperature, 20-40 degrees C. The highest DMSO-degradation activity was obtained at 30 degrees C, and 0.64 mM (50 mg/l) DMSO was completely degraded to sulfate ion with 100% molar conversion ratio within only 15 min. Furthermore, to examine whether WU-K217 would be useful for the removal of DMSO contained in wastewater exhausted in large amounts, continuous degradation of DMSO was examined. When 0.64 mM DMSO was added to the resting cells periodically at 15-min intervals, DMSO was completely degraded to sulfate ion without any decrease of the degradation activity at least during the twelve times of DMSO addition. PMID:16233269

  9. The nitric-oxide reductase from Paracoccus denitrificans uses a single specific proton pathway.

    PubMed

    ter Beek, Josy; Krause, Nils; Reimann, Joachim; Lachmann, Peter; Ädelroth, Pia

    2013-10-18

    The NO reductase from Paracoccus denitrificans reduces NO to N2O (2NO + 2H(+) + 2e(-) → N2O + H2O) with electrons donated by periplasmic cytochrome c (cytochrome c-dependent NO reductase; cNOR). cNORs are members of the heme-copper oxidase superfamily of integral membrane proteins, comprising the O2-reducing, proton-pumping respiratory enzymes. In contrast, although NO reduction is as exergonic as O2 reduction, there are no protons pumped in cNOR, and in addition, protons needed for NO reduction are derived from the periplasmic solution (no contribution to the electrochemical gradient is made). cNOR thus only needs to transport protons from the periplasm into the active site without the requirement to control the timing of opening and closing (gating) of proton pathways as is needed in a proton pump. Based on the crystal structure of a closely related cNOR and molecular dynamics simulations, several proton transfer pathways were suggested, and in principle, these could all be functional. In this work, we show that residues in one of the suggested pathways (denoted pathway 1) are sensitive to site-directed mutation, whereas residues in the other proposed pathways (pathways 2 and 3) could be exchanged without severe effects on turnover activity with either NO or O2. We further show that electron transfer during single-turnover reduction of O2 is limited by proton transfer and can thus be used to study alterations in proton transfer rates. The exchange of residues along pathway 1 showed specific slowing of this proton-coupled electron transfer as well as changes in its pH dependence. Our results indicate that only pathway 1 is used to transfer protons in cNOR.

  10. Description of Rhodanobacter denitrificans sp. nov., isolated from nitrate-rich zones of a contaminated aquifer

    SciTech Connect

    Prakash, Om; Green, Stefan; Jasrotia, Puja; Overholt, Will; Canion, Andy; Watson, David B; Brooks, Scott C; Kostka,

    2012-01-01

    Bacterial strains 2APBS1T and 116-2 were isolated from the subsurface of a nuclear legacy waste site where sediments are co-contaminated with large amounts of acidity, nitrate, metal radionuclides and other heavy metals. A combination of physiological and genetic assays indicated that these strains represent the first members of the Rhodanobacter genus shown to be capable of complete denitrification. Cells of strain 2APBS1T and 116-2 were Gram negative, non-spore-forming, rods, 3-5 micro;m long and 0.25-0.5 m in diameter. The isolates were facultative anaerobes, and had temperature and pH optima for growth at 30 C and pH 6.5, respectively, and could tolerate up to 2.0 % NaCl, though growth improved in its absence. Strains 2APBS1T and 116-2 contained fatty acid profiles and 100 % Q-8 ubiquinone, that are characteristic features of the genus Rhodanobacter. Although strains 2APBS1T and 116-2 share high SSU rRNA gene sequence similarity to R. thiooxydans (>99%), DNA-DNA hybridization values were substantially below the 70% threshold used to designate novel species. Thus, based on genotypic, phylogenetic, chemotaxonomic and physiological differences, strains 2APBS1T and 116-2 are considered to represent a novel species of the genus Rhodanobacter, for which the name Rhodanobacter denitrificans sp. nov is proposed. The type strain is 2APBS1T (=DSM 23569T =JCM 17641T). Strain 116-2 (=DSM 24678 = JCM 17642) is a reference strain.

  11. Description of Rhodanobacter denitrificans sp. nov., isolated from nitrate-rich zones of a contaminated aquifer

    SciTech Connect

    Prakash, Om; Green, Stefan; Jasrotia, Puja; Overholt, Will; Canion, Andy; Watson, David B; Brooks, Scott C; Kostka, Joel

    2011-01-01

    Bacterial strains 2APBS1T and 116-2 were isolated from the subsurface of a nuclear legacy waste site where sediments are co-contaminated with large amounts of acidity, nitrate, metal radionuclides and other heavy metals. A combination of physiological and genetic assays indicated that these strains represent the first members of the Rhodanobacter genus shown to be capable of complete denitrification. Cells of strain 2APBS1T and 116-2 were Gram negative, non-spore-forming, rods, 3-5 micro;m long and 0.25-0.5 m in diameter. The isolates were facultative anaerobes, and had temperature and pH optima for growth at 30 C and pH 6.5, respectively, and could tolerate up to 2.0 % NaCl, though growth improved in its absence. Strains 2APBS1T and 116-2 contained fatty acid profiles and 100 % Q-8 ubiquinone, that are characteristic features of the genus Rhodanobacter. Although strains 2APBS1T and 116-2 share high SSU rRNA gene sequence similarity to R. thiooxydans (>99%), DNA-DNA hybridization values were substantially below the 70% threshold used to designate novel species. Thus, based on genotypic, phylogenetic, chemotaxonomic and physiological differences, strains 2APBS1T and 116-2 are considered to represent a novel species of the genus Rhodanobacter, for which the name Rhodanobacter denitrificans sp. nov is proposed. The type strain is 2APBS1T (=DSM 23569T =JCM 17641T). Strain 116-2 (=DSM 24678 = JCM 17642) is a reference strain.

  12. Brevundimonas denitrificans sp. nov., a denitrifying bacterium isolated from deep subseafloor sediment.

    PubMed

    Tsubouchi, Taishi; Koyama, Sumihiro; Mori, Kozue; Shimane, Yasuhiro; Usui, Keiko; Tokuda, Maki; Tame, Akihiro; Uematsu, Katsuyuki; Maruyama, Tadashi; Hatada, Yuji

    2014-11-01

    A novel Gram-stain-negative, aerobic, heterotrophic, stalked and capsulated bacterium with potential denitrification ability, designated strain TAR-002(T), was isolated from deep seafloor sediment in Japan. Colonies lacked lustre, and were viscous and translucent white. The ranges of temperature, pH and salt concentration for growth were 8-30 °C, pH 6.0-10.0 and 1-3% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences confirmed that strain TAR-002(T) belongs to the genus Brevundimonas of the class Alphaproteobacteria. Levels of similarity between the 16S rRNA gene sequence of strain TAR-002(T) and those of the type strains of species of the genus Brevundimonas were 93.5-98.9%; the most closely related species was Brevundimonas basaltis. In DNA-DNA hybridization assays between strain TAR-002(T) and its phylogenetic neighbours, Brevundimonas lenta DS-18(T), B. basaltis J22(T), Brevundimonas subvibrioides ATCC 15264(T) and Brevundimonas alba DSM 4736(T), mean hybridization levels were 6.4-27.7%. The G+C content of strain TAR-002(T) was 70.3 mol%. Q-10 was the major respiratory isoprenoid quinone. The major fatty acids were C(18:1)ω7c and C(16:0), and the presence of 1,2-di-O-acyl-3-O-[D-glucopyranosyl-(1 → 4)-α-D-glucopyranuronosyl]glycerol (DGL) indicates the affiliation of strain TAR-002(T) with the genus Brevundimonas. On the basis of biological characteristics and 16S rRNA gene sequence comparisons, strain TAR-002(T) is considered to represent a novel species of the genus Brevundimonas, for which the name Brevundimonas denitrificans sp. nov. is proposed; the type strain is TAR-002(T) ( =NBRC 110107(T) =CECT 8537(T)).

  13. Ferrovibrio denitrificans gen. nov., sp. nov., a novel neutrophilic facultative anaerobic Fe(II)-oxidizing bacterium.

    PubMed

    Sorokina, Anna Y; Chernousova, Elena Y; Dubinina, Galina A

    2012-10-01

    A neutrophilic Fe(II)-oxidizing bacterium was isolated from the redox zone of a low-salinity spring in Krasnodar krai (Russia), at the FeS-Fe(OH)(3) interface deposited at the sediment surface. The cells of strain Sp-1 were short, thin motile vibrioids with one polar flagellum dividing by binary fission. The optimal values and ranges for pH and temperature were pH 6.2 (5.5-8) and 35 °C (5-45 °C), respectively. The organism was a facultative anaerobe. Strain Sp-1 was capable of organotrophic, lithoheterotrophic and mixotrophic growth with Fe(II) as an electron donor. The denitrification chain was 'disrupted'. Oxidation of Fe(II) was coupled to reduction of NO3 - to NO2 - or of N(2) O to N(2) , as well as under microaerobic conditions, with O(2) as an electron acceptor. The DNA G+C content was 64.2 mol%. According to the results of phylogenetic analysis, the strain was 10.6-12% remote from the closest relatives, members of the genera Sneathiella, Inquilinus, Oceanibaculum and Phaeospirillum within the Alphaproteobacteria. Based on its morphological, physiological and taxonomic characteristics, together with the results of phylogenetic analysis, strain Sp-1 is described as a member of a new genus Ferrovibrio gen. nov., with the type species Ferrovibrio denitrificans sp. nov. and the type strain Sp-1(T) (= LMG 25817(T)  = VKM B-2673(T) ). PMID:22765162

  14. Involvement in Denitrification is Beneficial to the Biofilm Lifestyle of Comamonas testosteroni: A Mechanistic Study and Its Environmental Implications.

    PubMed

    Wu, Yichao; Shukal, Sudha; Mukherjee, Manisha; Cao, Bin

    2015-10-01

    Comamonas is one of the most abundant microorganisms in biofilm communities driving wastewater treatment. Little has been known about the role of this group of organisms and their biofilm mode of life. In this study, using Comamonas testosteroni as a model organism, we demonstrated the involvement of Comamonas biofilms in denitrification under bulk aerobic conditions and elucidated the influence of nitrate respiration on its biofilm lifestyle. Our results showed that C. testosteroni could use nitrate as the sole electron acceptor for anaerobic growth. Under bulk aerobic condition, biofilms of C. testosteroni were capable of reducing nitrate, and intriguingly, nitrate reduction significantly enhanced viability of the biofilm-cells and reduced cell detachment from the biofilms. Nitrate respiration was further shown to play an essential role in maintaining high cell viability in the biofilms. RNA-seq analysis, quantitative polymerase chain reaction, and liquid chromatography-mass spectrometry revealed a higher level of bis(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP) in cells respiring on nitrate than those grown aerobically (1.3 × 10(-4) fmol/cell vs 7.9 × 10(-6) fmol/cell; P < 0.01). C-di-GMP is one universal signaling molecule that regulates the biofilm mode of life, and a higher c-di-GMP concentration reduces cell detachment from biofilms. Taking these factors together, this study reveals that nitrate reduction occurs in mature biofilms of C. testosteroni under bulk aerobic conditions, and the respiratory reduction of nitrate is beneficial to the biofilm lifestyle by providing more metabolic energy to maintain high viability and a higher level of c-di-GMP to reduce cell detachment. PMID:26327221

  15. Direct sensing and signal transduction during bacterial chemotaxis toward aromatic compounds in Comamonas testosteroni.

    PubMed

    Huang, Zhou; Ni, Bin; Jiang, Cheng-Ying; Wu, Yu-Fan; He, Yun-Zhe; Parales, Rebecca E; Liu, Shuang-Jiang

    2016-07-01

    Micro-organisms sense and chemotactically respond to aromatic compounds. Although the existence of chemoreceptors that bind to aromatic attractants and subsequently trigger chemotaxis have long been speculated, such a chemoreceptor has not been demonstrated. In this report, we demonstrated that the chemoreceptor MCP2901 from Comamonas testosteroni CNB-1 binds to aromatic compounds and initiates downstream chemotactic signaling in addition to its ability to trigger chemotaxis via citrate binding. The function of gene MCP2901 was investigated by genetic deletion from CNB-1 and genetic complementation of the methyl-accepting chemotaxis protein (MCP)-null mutant CNB-1Δ20. Results showed that the expression of MCP2901 in the MCP-null mutant restored chemotaxis toward nine tested aromatic compounds and nine carboxylic acids. Isothermal titration calorimetry (ITC) analyses demonstrated that the ligand-binding domain of MCP2901 (MCP2901LBD) bound to citrate, and weakly to gentisate and 4-hydroxybenzoate. Additionally, ITC assays indicated that MCP2901LBD bound strongly to 2,6-dihydroxybenzoate and 2-hydroxybenzoate, which are isomers of gentisate and 4-hydroxybenzoate respectively that are not metabolized by CNB-1. Agarose-in-plug and capillary assays showed that these two molecules serve as chemoattractants for CNB-1. Through constructing membrane-like MCP2901-inserted Nanodiscs and phosphorelay activity assays, we demonstrated that 2,6-dihydroxybenzoate and 2-hydroxybenzoate altered kinase activity of CheA. This is the first evidence of an MCP binding to an aromatic molecule and triggering signal transduction for bacterial chemotaxis.

  16. Molecular analysis of the poly(3-hydroxyalkanoate) synthase gene from a methylotrophic bacterium, Paracoccus denitrificans.

    PubMed Central

    Ueda, S; Yabutani, T; Maehara, A; Yamane, T

    1996-01-01

    A 3.6-kb EcoRI-SalI fragment of Paracoccus denitrificans DNA hybridized with a DNA probe carrying the poly(3-hydroxyalkanoate) (PHA) synthase gene (phaC) of Alcaligenes eutrophus. Nucleotide sequence analysis of this region showed the presence of a 1,872-bp open reading frame (ORF), which corresponded to a polypeptide with a molecular weight of 69,537. Upstream of the ORF, a promoter-like sequence was found. Escherichia coli carrying the fusion gene between lacZ and the ORF accumulated a level of poly(3-hydroxybutyrate) that was as much as 20 wt% of the cell dry weight in the presence of beta-ketothiolase and acetoacetylcoenzyme A reductase genes of A. eutrophus. The ORF was designated phaCPd. A plasmid vector carrying the phaCPd'-'lacZ fusion gene downstream of the promoter-like sequence expressed beta-galactosidase activity in P. denitrificans. When a multicopy and broad-host-range vector carrying the ORF along with the promoter-like sequence was introduced into P. denitrificans, the PHA content in the cells increased by twofold compared with cells carrying only a vector sequence. PMID:8550512

  17. Molecular analysis of the poly(3-hydroxyalkanoate) synthase gene from a methylotrophic bacterium, Paracoccus denitrificans.

    PubMed

    Ueda, S; Yabutani, T; Maehara, A; Yamane, T

    1996-02-01

    A 3.6-kb EcoRI-SalI fragment of Paracoccus denitrificans DNA hybridized with a DNA probe carrying the poly(3-hydroxyalkanoate) (PHA) synthase gene (phaC) of Alcaligenes eutrophus. Nucleotide sequence analysis of this region showed the presence of a 1,872-bp open reading frame (ORF), which corresponded to a polypeptide with a molecular weight of 69,537. Upstream of the ORF, a promoter-like sequence was found. Escherichia coli carrying the fusion gene between lacZ and the ORF accumulated a level of poly(3-hydroxybutyrate) that was as much as 20 wt% of the cell dry weight in the presence of beta-ketothiolase and acetoacetylcoenzyme A reductase genes of A. eutrophus. The ORF was designated phaCPd. A plasmid vector carrying the phaCPd'-'lacZ fusion gene downstream of the promoter-like sequence expressed beta-galactosidase activity in P. denitrificans. When a multicopy and broad-host-range vector carrying the ORF along with the promoter-like sequence was introduced into P. denitrificans, the PHA content in the cells increased by twofold compared with cells carrying only a vector sequence.

  18. Sulfurimonas paralvinellae sp. nov., a novel mesophilic, hydrogen- and sulfur-oxidizing chemolithoautotroph within the Epsilonproteobacteria isolated from a deep-sea hydrothermal vent polychaete nest, reclassification of Thiomicrospira denitrificans as Sulfurimonas denitrificans comb. nov. and emended description of the genus Sulfurimonas.

    PubMed

    Takai, Ken; Suzuki, Masae; Nakagawa, Satoshi; Miyazaki, Masayuki; Suzuki, Yohey; Inagaki, Fumio; Horikoshi, Koki

    2006-08-01

    A novel mesophilic bacterium, strain GO25(T), was isolated from a nest of hydrothermal vent polychaetes, Paralvinella sp., at the Iheya North field in the Mid-Okinawa Trough. Cells were motile short rods with a single polar flagellum. Growth was observed between 4 and 35 degrees C (optimum 30 degrees C; 13-16 h doubling time) and between pH 5.4 and 8.6 (optimum pH 6.1). The isolate was a facultatively anaerobic chemolithoautotroph capable of growth using molecular hydrogen, elemental sulfur or thiosulfate as the sole energy source, carbon dioxide as the sole carbon source, ammonium or nitrate as the sole nitrogen source and elemental sulfur, thiosulfate or yeast extract as the sole sulfur source. Strain GO25(T) represents the first deep-sea epsilonproteobacterium capable of growth by both hydrogen and sulfur oxidation. Nitrate or molecular oxygen (up to 10 % partial pressure) could serve as the sole electron acceptor to support growth. Metabolic products of nitrate reduction shifted in response to the electron donor provided. The G+C content of genomic DNA was 37.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the novel isolate belonged to the genus Sulfurimonas and was most closely related to Sulfurimonas autotrophica OK10(T) (96.3 % sequence similarity). DNA-DNA hybridization demonstrated that the novel isolate could be differentiated genotypically from Sulfurimonas autotrophica OK10(T). On the basis of the physiological and molecular properties of the novel isolate, the name Sulfurimonas paralvinellae sp. nov. is proposed, with strain GO25(T) (=JCM 13212(T)=DSM 17229(T)) as the type strain. Thiomicrospira denitrificans DSM 1251(T) (=ATCC 33889(T)) is phylogenetically associated with Sulfurimonas autotrophica OK10(T) and Sulfurimonas paralvinellae GO25(T). Based on the phylogenetic relationship between Thiomicrospira denitrificans DSM 1251(T), Sulfurimonas autotrophica OK10(T) and Sulfurimonas paralvinellae GO25(T), we propose the

  19. 3-Hydroxybutyrate oligomer hydrolase and 3-hydroxybutyrate dehydrogenase participate in intracellular polyhydroxybutyrate and polyhydroxyvalerate degradation in Paracoccus denitrificans.

    PubMed

    Lu, Jing; Takahashi, Akira; Ueda, Shunsaku

    2014-02-01

    Genes encoding 3-hydroxybutyrate oligomer hydrolase (PhaZc) and 3-hydroxybutyrate dehydrogenase (Hbd) were isolated from Paracoccus denitrificans. PhaZc and Hbd were overproduced as His-tagged proteins in Escherichia coli and purified by affinity and gel filtration chromatography. Purified His-tagged proteins had molecular masses of 31 kDa and 120 kDa (a tetramer of 29-kDa subunits). The His-tagged PhaZc hydrolyzed not only 3-hydroxybutyrate oligomers but also 3-hydroxyvalerate oligomers. The His-tagged Hbd catalyzed the dehydrogenation of 3-hydroxyvalerate as well as 3-hydroxybutyrate. When both enzymes were included in the same enzymatic reaction system with 3-hydroxyvalerate dimer, sequential reactions occurred, suggesting that PhaZc and Hbd play an important role in the intracellular degradation of poly(3-hydroxyvalerate). When the phaZc gene was disrupted in P. denitrificans by insertional inactivation, the mutant strain lost PhaZc activity. When the phaZc-disrupted P. denitrificans was complemented with phaZc, PhaZc activity was restored. These results suggest that P. denitrificans carries a single phaZc gene. Disruption of the phaZc gene in P. denitrificans affected the degradation rate of PHA. PMID:24271169

  20. Growth and nitrite and nitrous oxide accumulation of Paracoccus denitrificans ATCC 19367 in the presence of selected pesticides.

    PubMed

    Sáez, Florentina; Pozo, Clementina; Gómez, Miguel Angel; Rodelas, Belén; Gónzalez-López, Jesús

    2003-09-01

    The effects of the application of eight pesticides (aldrin, lindane, dimetoate, methylparathion, methidation, atrazine, simazine, and captan) on growth, respiratory activity (as CO2 production), denitrifying activity (as N2O released), and nitrite accumulation in the culture medium by Paracoccus denitrificans strain ATCC 19367 were studied. The fungicide captan totally inhibited growth and biological activity of P. denitrificans, while the rest of the tested pesticides delayed the growth and CO2 release of P. denitrificans but did not drastically affect the bacterial growth or respiratory capacity after 96 h of culture. The denitrifying activity of P. denitrificans ATCC 19367 (as N2O released) was negatively affected by all tested pesticides. The release of N2O was strongly inhibited by several organochlorinated and organophosphorated insecticides (aldrin, lindane, dimetoate, and methidation), which led to high accumulation of nitrite in the surrounding medium. Atrazine decreased N2O release after 48 h of culture because of negative effects on growth, and methylparathion and simazine delayed the onset of N2O release by P. denitrificans. These three pesticides reduced the accumulation of NO2- compared to unamended control cultures. PMID:12959522

  1. Identification of the intracellular polyhydroxyalkanoate depolymerase gene of Paracoccus denitrificans and some properties of the gene product.

    PubMed

    Gao, D; Maehara, A; Yamane, T; Ueda, S

    2001-03-15

    Paracoccus denitrificans degraded poly(3-hydroxybutyrate) (PHB) in the cells under carbon source starvation. Intracellular poly(3-hydroxyalkanoate) (PHA) depolymerase gene (phaZ) was identified near the PHA synthase gene (phaC) of P. denitrificans. Cell extract of Escherichia coli carrying lacZ--phaZ fusion gene degraded protease-treated PHB granules. Reaction products were thought to be mainly D(--)-3-hydroxybutyrate (3HB) dimer and 3HB oligomer. Diisopropylfluorophosphonate and Triton X-100 exhibited an inhibitory effect on the degradation of PHB granules. When cell extract of the recombinant E. coli was used, Mg(2+) ion inhibited PHB degradation. However, the inhibitory effect by Mg(2+) ion was not observed using the cell extract of P. denitrificans.

  2. Biodegradation of propyzamide by Comamonas testosteroni W1 and cloning of the propyzamide hydrolase gene camH.

    PubMed

    Zhao, Baiping; Hua, Xiude; Wang, Fei; Dong, Weiliang; Li, Zhoukun; Yang, Yu; Cui, Zhongli; Wang, Minghua

    2015-03-01

    Propyzamide is a widely used benzamide herbicide for controlling weeds in lettuce, soybeans, cotton and other crops. An efficient propyzamide-degrading strain W1 was firstly isolated from activated sludge and identified as Comamonas testosteroni. A metabolite of propyzamide by strain W1 was firstly identified. The novel gene camH encoding a hydrolase that catalyzed the amide bond cleavage of propyzamide was cloned from strain W1. The gene contained an open reading frame of 1452 bp, the deduced amino acid sequence showed low identity with other amidases. The recombinant enzyme CamH was expressed in Escherichia coli BL21 and purified. CamH displayed the highest activity at 30°C and pH 8.0 with propyzamide as the substrate. These results provide important knowledge on the fate of propyzamide in the biodegradation, and elucidate the biodegradation mechanism of propyzamide by the strain W1.

  3. Bioaugmentation of activated sludge with elemental sulfur producing strain Thiopseudomonas denitrificans X2 against nitrate shock load.

    PubMed

    Tan, Wenbo; Huang, Cong; Chen, Chuan; Liang, Bin; Wang, Aijie

    2016-11-01

    The sulfide and nitrogen compounds in wastewaters are toxic and cause a serious environmental problem. Thiopseudomonas denitrificans X2, which is the type species of a novel genus Thiopseudomonas was used for bioaugmentation. It oxidized sulfide and acetate with nitrate, and generated elemental sulfur that could be recovered as resource. The generation rate of elemental sulfur was enhanced significantly by the bioaugmentation under the condition of excessive nitrate feeding. The inoculums survived and worked actively in the activated sludge system as the dominant population. Thiopseudomonas denitrificans X2 could be applied to wastewater treatment and resource recovery simultaneously. PMID:27590576

  4. The genome sequence of the obligately chemolithoautotrophic, facultatively anaerobic bacterium Thiobacillus denitrificans.

    PubMed

    Beller, Harry R; Chain, Patrick S G; Letain, Tracy E; Chakicherla, Anu; Larimer, Frank W; Richardson, Paul M; Coleman, Matthew A; Wood, Ann P; Kelly, Donovan P

    2006-02-01

    The complete genome sequence of Thiobacillus denitrificans ATCC 25259 is the first to become available for an obligately chemolithoautotrophic, sulfur-compound-oxidizing, beta-proteobacterium. Analysis of the 2,909,809-bp genome will facilitate our molecular and biochemical understanding of the unusual metabolic repertoire of this bacterium, including its ability to couple denitrification to sulfur-compound oxidation, to catalyze anaerobic, nitrate-dependent oxidation of Fe(II) and U(IV), and to oxidize mineral electron donors. Notable genomic features include (i) genes encoding c-type cytochromes totaling 1 to 2 percent of the genome, which is a proportion greater than for almost all bacterial and archaeal species sequenced to date, (ii) genes encoding two [NiFe]hydrogenases, which is particularly significant because no information on hydrogenases has previously been reported for T. denitrificans and hydrogen oxidation appears to be critical for anaerobic U(IV) oxidation by this species, (iii) a diverse complement of more than 50 genes associated with sulfur-compound oxidation (including sox genes, dsr genes, and genes associated with the AMP-dependent oxidation of sulfite to sulfate), some of which occur in multiple (up to eight) copies, (iv) a relatively large number of genes associated with inorganic ion transport and heavy metal resistance, and (v) a paucity of genes encoding organic-compound transporters, commensurate with obligate chemolithoautotrophy. Ultimately, the genome sequence of T. denitrificans will enable elucidation of the mechanisms of aerobic and anaerobic sulfur-compound oxidation by beta-proteobacteria and will help reveal the molecular basis of this organism's role in major biogeochemical cycles (i.e., those involving sulfur, nitrogen, and carbon) and groundwater restoration.

  5. Identification of periplasmic nitrate reductase Mo(V) EPR signals in intact cells of Paracoccus denitrificans.

    PubMed

    Sears, H J; Bennett, B; Spiro, S; Thomson, A J; Richardson, D J

    1995-08-15

    EPR spectroscopy has been successfully used to detect signals due to molybdenum (V) and ferric iron in intact cells of aerobically grown Paracoccus denitrificans. The signals are ascribed to the catalytic molybdenum centre and to the haem iron of the periplasmic nitrate reductase. These signals are absent from a mutant strain deficient in this enzyme. The Mo(V) signal is due to the High-g Split species which has been well characterized in the purified enzyme. This confirms that the High-g Split is the physiologically relevant signal of a number observed in the previous work on the purified enzyme. PMID:7646461

  6. Proteomic analysis of nitrate-dependent acetone degradation by Alicycliphilus denitrificans strain BC.

    PubMed

    Oosterkamp, Margreet J; Boeren, Sjef; Atashgahi, Siavash; Plugge, Caroline M; Schaap, Peter J; Stams, Alfons J M

    2015-06-01

    Alicycliphilus denitrificans strain BC grows anaerobically on acetone with nitrate as electron acceptor. Comparative proteomics of cultures of A. denitrificans strain BC grown on either acetone or acetate with nitrate was performed to study the enzymes involved in the acetone degradation pathway. In the proposed acetone degradation pathway, an acetone carboxylase converts acetone to acetoacetate, an AMP-dependent synthetase/ligase converts acetoacetate to acetoacetyl-CoA, and an acetyl-CoA acetyltransferase cleaves acetoacetyl-CoA to two acetyl-CoA. We also found a putative aldehyde dehydrogenase associated with acetone degradation. This enzyme functioned as a β-hydroxybutyrate dehydrogenase catalyzing the conversion of surplus acetoacetate to β-hydroxybutyrate that may be converted to the energy and carbon storage compound, poly-β-hydroxybutyrate. Accordingly, we confirmed the formation of poly-β-hydroxybutyrate in acetone-grown cells of strain BC. Our findings provide insight in nitrate-dependent acetone degradation that is activated by carboxylation of acetone. This will aid studies of similar pathways found in other microorganisms degrading acetone with nitrate or sulfate as electron acceptor.

  7. Nitrogen, carbon, and sulfur isotopic change during heterotrophic (Pseudomonas aureofaciens) and autotrophic (Thiobacillus denitrificans) denitrification reactions.

    PubMed

    Hosono, Takahiro; Alvarez, Kelly; Lin, In-Tian; Shimada, Jun

    2015-12-01

    In batch culture experiments, we examined the isotopic change of nitrogen in nitrate (δ(15)NNO3), carbon in dissolved inorganic carbon (δ(13)CDIC), and sulfur in sulfate (δ(34)SSO4) during heterotrophic and autotrophic denitrification of two bacterial strains (Pseudomonas aureofaciens and Thiobacillus denitrificans). Heterotrophic denitrification (HD) experiments were conducted with trisodium citrate as electron donor, and autotrophic denitrification (AD) experiments were carried out with iron disulfide (FeS2) as electron donor. For heterotrophic denitrification experiments, a complete nitrate reduction was accomplished, however bacterial denitrification with T. denitrificans is a slow process in which, after seventy days nitrate was reduced to 40% of the initial concentration by denitrification. In the HD experiment, systematic change of δ(13)CDIC (from -7.7‰ to -12.2‰) with increase of DIC was observed during denitrification (enrichment factor εN was -4.7‰), suggesting the contribution of C of trisodium citrate (δ(13)C=-12.4‰). No SO4(2-) and δ(34)SSO4 changes were observed. In the AD experiment, clear fractionation of δ(13)CDIC during DIC consumption (εC=-7.8‰) and δ(34)SSO4 during sulfur use of FeS2-S (around 2‰), were confirmed through denitrification (εN=-12.5‰). Different pattern in isotopic change between HD and AD obtained on laboratory-scale are useful to recognize the type of denitrification occurring in the field. PMID:26529303

  8. Nitrogen, carbon, and sulfur isotopic change during heterotrophic (Pseudomonas aureofaciens) and autotrophic (Thiobacillus denitrificans) denitrification reactions

    NASA Astrophysics Data System (ADS)

    Hosono, Takahiro; Alvarez, Kelly; Lin, In-Tian; Shimada, Jun

    2015-12-01

    In batch culture experiments, we examined the isotopic change of nitrogen in nitrate (δ15NNO3), carbon in dissolved inorganic carbon (δ13CDIC), and sulfur in sulfate (δ34SSO4) during heterotrophic and autotrophic denitrification of two bacterial strains (Pseudomonas aureofaciens and Thiobacillus denitrificans). Heterotrophic denitrification (HD) experiments were conducted with trisodium citrate as electron donor, and autotrophic denitrification (AD) experiments were carried out with iron disulfide (FeS2) as electron donor. For heterotrophic denitrification experiments, a complete nitrate reduction was accomplished, however bacterial denitrification with T. denitrificans is a slow process in which, after seventy days nitrate was reduced to 40% of the initial concentration by denitrification. In the HD experiment, systematic change of δ13CDIC (from - 7.7‰ to - 12.2‰) with increase of DIC was observed during denitrification (enrichment factor εN was - 4.7‰), suggesting the contribution of C of trisodium citrate (δ13C = - 12.4‰). No SO42 - and δ34SSO4 changes were observed. In the AD experiment, clear fractionation of δ13CDIC during DIC consumption (εC = - 7.8‰) and δ34SSO4 during sulfur use of FeS2-S (around 2‰), were confirmed through denitrification (εN = - 12.5‰). Different pattern in isotopic change between HD and AD obtained on laboratory-scale are useful to recognize the type of denitrification occurring in the field.

  9. Nitrogen, carbon, and sulfur isotopic change during heterotrophic (Pseudomonas aureofaciens) and autotrophic (Thiobacillus denitrificans) denitrification reactions.

    PubMed

    Hosono, Takahiro; Alvarez, Kelly; Lin, In-Tian; Shimada, Jun

    2015-12-01

    In batch culture experiments, we examined the isotopic change of nitrogen in nitrate (δ(15)NNO3), carbon in dissolved inorganic carbon (δ(13)CDIC), and sulfur in sulfate (δ(34)SSO4) during heterotrophic and autotrophic denitrification of two bacterial strains (Pseudomonas aureofaciens and Thiobacillus denitrificans). Heterotrophic denitrification (HD) experiments were conducted with trisodium citrate as electron donor, and autotrophic denitrification (AD) experiments were carried out with iron disulfide (FeS2) as electron donor. For heterotrophic denitrification experiments, a complete nitrate reduction was accomplished, however bacterial denitrification with T. denitrificans is a slow process in which, after seventy days nitrate was reduced to 40% of the initial concentration by denitrification. In the HD experiment, systematic change of δ(13)CDIC (from -7.7‰ to -12.2‰) with increase of DIC was observed during denitrification (enrichment factor εN was -4.7‰), suggesting the contribution of C of trisodium citrate (δ(13)C=-12.4‰). No SO4(2-) and δ(34)SSO4 changes were observed. In the AD experiment, clear fractionation of δ(13)CDIC during DIC consumption (εC=-7.8‰) and δ(34)SSO4 during sulfur use of FeS2-S (around 2‰), were confirmed through denitrification (εN=-12.5‰). Different pattern in isotopic change between HD and AD obtained on laboratory-scale are useful to recognize the type of denitrification occurring in the field.

  10. Structure of the Membrane-intrinsic Nitric Oxide Reductase from Roseobacter denitrificans.

    PubMed

    Crow, Allister; Matsuda, Yuji; Arata, Hiroyuki; Oubrie, Arthur

    2016-06-14

    Membrane-intrinsic nitric oxide reductases (NORs) are key components of bacterial denitrification pathways with a close evolutionary relationship to the cytochrome oxidase (COX) complex found in aerobic respiratory chains. A key distinction between COX and NOR is the identity of the metal directly opposite heme b3 within the active site. In NOR, this metal is iron (FeB), whereas in COX, it is copper (CuB). The purified NOR of Roseobacter denitrificans contains copper and has modest oxidase activity, raising the possibility that a COX-like active site might have independently arisen within the context of a NOR-like protein scaffold. Here we present the crystal structure of the Roseobacter denitrificans NorBC complex and anomalous scattering experiments probing the identity of each metal center. Our results refute the hypothesis that copper occupies the active site and instead reveal a new metal center in the small subunit not seen in any other NOR or COX. PMID:27185533

  11. Purification, characterization and crystallization of the F-ATPase from Paracoccus denitrificans.

    PubMed

    Morales-Rios, Edgar; Watt, Ian N; Zhang, Qifeng; Ding, Shujing; Fearnley, Ian M; Montgomery, Martin G; Wakelam, Michael J O; Walker, John E

    2015-09-01

    The structures of F-ATPases have been determined predominantly with mitochondrial enzymes, but hitherto no F-ATPase has been crystallized intact. A high-resolution model of the bovine enzyme built up from separate sub-structures determined by X-ray crystallography contains about 85% of the entire complex, but it lacks a crucial region that provides a transmembrane proton pathway involved in the generation of the rotary mechanism that drives the synthesis of ATP. Here the isolation, characterization and crystallization of an integral F-ATPase complex from the α-proteobacterium Paracoccus denitrificans are described. Unlike many eubacterial F-ATPases, which can both synthesize and hydrolyse ATP, the P. denitrificans enzyme can only carry out the synthetic reaction. The mechanism of inhibition of its ATP hydrolytic activity involves a ζ inhibitor protein, which binds to the catalytic F₁-domain of the enzyme. The complex that has been crystallized, and the crystals themselves, contain the nine core proteins of the complete F-ATPase complex plus the ζ inhibitor protein. The formation of crystals depends upon the presence of bound bacterial cardiolipin and phospholipid molecules; when they were removed, the complex failed to crystallize. The experiments open the way to an atomic structure of an F-ATPase complex.

  12. Production, isolation and characterization of monoclonal antibodies to cytochromes c of beef heart and Paracoccus denitrificans.

    PubMed

    Kuo, L M; Davies, H C

    1983-08-01

    Hybridoma cell lines secreting monoclonal antibodies which bind beef heart cytochrome c or Paracoccus denitrificans cytochrome c have been produced using spleen cells from BALB/c mice immunized with cytochrome c. Immunization was performed with either the native cytochrome c, succinylated hemocyanin-conjugated cytochrome c, or beef heart cytochrome c polymerized with glutaraldehyde. Of 10 such fusions, the hybridization frequency ranged from 0 to 42%. The cell fusion efficiency, the possible factors involved in the cell fusion efficiency and the frequency of antibody producing hybridomas are described. The percentage of hybridomas positive for anti-cytochrome c antibody production as screened for by radioimmunoassay or ELISA was 2%. Of the antibodies from 12 hybridoma cell lines which resulted from 10 fusions, three were specific to beef heart cytochrome c, another three were specific to P. denitrificans cytochrome c, and the remainder reacted with both cytochromes c. These groups of monoclonal antibodies react to different sets of sites on these two cytochromes c. The monoclonal antibodies from ten representative clones have been isolated and characterized by different methods.

  13. Co-Cultures of Pseudomonas aeruginosa and Roseobacter denitrificans Reveal Shifts in Gene Expression Levels Compared to Solo Cultures

    PubMed Central

    Conway, Crystal A.; Esiobu, Nwadiuto; Lopez, Jose V.

    2012-01-01

    Consistent biosynthesis of desired secondary metabolites (SMs) from pure microbial cultures is often unreliable. In a proof-of-principle study to induce SM gene expression and production, we describe mixed “co-culturing” conditions and monitoring of messages via quantitative real-time PCR (qPCR). Gene expression of model bacterial strains (Pseudomonas aeruginosa PAO1 and Roseobacter denitrificans Och114) was analyzed in pure solo and mixed cocultures to infer the effects of interspecies interactions on gene expression in vitro, Two P. aeruginosa genes (PhzH coding for portions of the phenazine antibiotic pathway leading to pyocyanin (PCN) and the RhdA gene for thiosulfate: cyanide sulfurtransferase (Rhodanese)) and two R. denitrificans genes (BetaLact for metallo-beta-lactamase and the DMSP gene for dimethylpropiothetin dethiomethylase) were assessed for differential expression. Results showed that R. denitrificans DMSP and BetaLact gene expression became elevated in a mixed culture. In contrast, P. aeruginosa co-cultures with R. denitrificans or a third species did not increase target gene expression above control levels. This paper provides insight for better control of target SM gene expression in vitro and bypass complex genetic engineering manipulations. PMID:22566756

  14. Achromobactor denitrificans SP1 produces pharmaceutically active 25C prodigiosin upon utilizing hazardous di(2-ethylhexyl)phthalate

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Achromobacter denitrificans SP1 isolated from soil sludge heavily contaminated with plastic waste produced a novel pharmaceutically-active 25C prodigiosin analog during growth in a simple mineral salt medium supplemented with hazardous di(2-ethylhexyl)phthalate (DEHP) blended PVC plastics (in situ) ...

  15. Bioremediation of Petrochemical Wastewater Containing BTEX Compounds by a New Immobilized Bacterium Comamonas sp. JB in Magnetic Gellan Gum.

    PubMed

    Jiang, Bei; Zhou, Zunchun; Dong, Ying; Wang, Bai; Jiang, Jingwei; Guan, Xiaoyan; Gao, Shan; Yang, Aifu; Chen, Zhong; Sun, Hongjuan

    2015-05-01

    In this study, we investigated the bioremediation of petrochemical wastewater containing BTEX compounds by immobilized Comamonas sp. JB cells. Three kinds of magnetic nanoparticles were evaluated as immobilization supports for strain JB. After comparison with Fe3O4 and a-Fe2O3 nanoparticles, r-Fe2O3 nanoparticle was selected as the optimal immobilization support. The highest biodegradation activity of r-Fe2O3-magnetically immobilized cells was obtained when the concentration of r-Fe2O3 nanoparticle was 120 mg L(-1). Additionally, the recycling experiments demonstrated that the degradation activity of r-Fe2O3-magnetically immobilized cells was still high and led to less toxicity than untreated wastewater during the eight recycles. qPCR suggested the concentration of strain JB in r-Fe2O3-magnetically immobilized cells was evidently increased after eight cycles of degradation experiments. These results supported developing efficient biocatalysts using r-Fe2O3-magnetically immobilized cells and provided a promising technique for improving biocatalysts used in the bioremediation of not only petrochemical wastewater but also other hazardous wastewater.

  16. Purification, structural elucidation and bioactivity of tryptophan containing diketopiperazines, from Comamonas testosteroni associated with a rhabditid entomopathogenic nematode against major human-pathogenic bacteria.

    PubMed

    Kumar, S Nishanth; Mohandas, C; Nambisan, Bala

    2014-03-01

    The cell free culture filtrate of a Comamonas testosteroni associated with an Entomopathogenic nematode (EPN), Rhabditis (Oscheius) sp. exhibited promising antimicrobial activity. The ethyl acetate extract of the bacterial culture filtrate was purified by silica gel column chromatography to obtain five diketopiperazines or cyclic dipeptides (DKP 1-5). The structure and absolute stereochemistry of the compounds were determined based on extensive spectroscopic analyses (HR-MS, (1)HNMR, (13)CNMR, (1)H-(1)H COSY, (1)H-(13)C HMBC) and Marfey's method. Based on the spectral data the compounds were identified as Cyclo-(L-Trp-L-Pro) (1), Cyclo-(L-Trp-L-Tyr) (2), Cyclo-(L-Trp-L-Ile) (3), Cyclo-(L-Trp-L-Leu) (4) and Cyclo-(L-Trp-L-Phe) (5), respectively. Three diketopiperazines (DKP 2, 3 and 5) were active against all the ten bacteria tested. The highest activity of 0.5μg/ml by Cyclo-(L-Trp-L-Phe) was recorded against Vibrio cholerae followed by Salmonella typhi (1 μg/ml) a human pathogen responsible for life threatening diseases like profuse watery diarrhea and typhoid or enteric fever. The activity of this compound against V. cholerae and S. typhi is more effective than ciprofloxacin and ampicillin, the standard antibiotics. Cyclo-(L-Trp-L-Phe) recorded significant antibacterial activity against all the test bacteria when compared to other compounds. Five diketopiperazines were active against all the test fungi and are more effective than bavistin the standard fungicide. Diketopiperazines recorded no cytotoxicity to FS normal fibroblast and VERO cells (African green monkey kidney) except DKP 3 and 4. To our best knowledge this is the first report of antimicrobial activity of the tryptophan containing diketopiperazines against the human pathogenic microbes. The production of cyclic dipeptides by C. testosteroni is also reported here for the first time. We conclude that the C. testosteroni is promising sources of natural bioactive secondary metabolites against human

  17. Nitric Oxide Is a Signal for NNR-Mediated Transcription Activation in Paracoccus denitrificans

    PubMed Central

    Van Spanning, Rob J. M.; Houben, Edith; Reijnders, Willem N. M.; Spiro, Stephen; Westerhoff, Hans V.; Saunders, Neil

    1999-01-01

    By using the ′lacZ gene, the activities of the nirI, nirS, and norC promoters were assayed in the wild type and in NNR-deficient mutants of Paracoccus denitrificans grown under various growth conditions. In addition, induction profiles of the three promoters in response to the presence of various nitrogenous oxides were determined. Transcription from the three promoters required the absence of oxygen and the presence both of the transcriptional activator NNR and of nitric oxide. The activity of the nnr promoter itself was halved after the cells had been switched from aerobic respiration to denitrification. This response was apparently not a result of autoregulation or of regulation by FnrP, since the nnr promoter was as active in the wild-type strain as it was in NNR- or FnrP-deficient mutants. PMID:10383987

  18. Purification and properties of D-(-)-3-hydroxybutyrate oligomer hydrolase of Paracoccus denitrificans.

    PubMed

    Ueda, Shunsaku; Sano, Konomi; Gao, Dai; Tomihari, Nao; Yamane, Tsuneo; Endo, Isao

    2002-01-10

    D-(-)-3-Hydroxybutyrate (3HB) oligomer hydrolase was purified from Paracoccus denitrificans. The enzyme was a monomeric protein with an approximate molecular mass of 31 kDa. The isoelectric point of the enzyme was 5.2. Optimum temperature and pH were 35-40 degrees C and 8.0, respectively. The enzyme activity was not affected by sulfhydryl reagents but strongly inhibited by serine proteinase inhibitors. Both 3HB trimer and 3HB dimer were hydrolyzed by the enzyme, indicating that the enzyme is not 3HB dimer hydrolase but 3HB oligomer hydrolase. para-Nitrophenyl esters of short-chain fatty acids were also hydrolyzed by the enzyme. 3HB dimer was hydrolyzed somewhat faster than 3HB trimer. The level of the enzyme activity was almost constant, irrespective of carbon sources for the bacterial growth and of the cultivation conditions.

  19. Kinetic properties of ATP sulfurylase and APS kinase from Thiobacillus denitrificans.

    PubMed

    Gay, Sean C; Fribourgh, Jennifer L; Donohoue, Paul D; Segel, Irwin H; Fisher, Andrew J

    2009-09-01

    The Thiobacillus denitrificans genome contains two sequences corresponding to ATP sulfurylase (Tbd_0210 and Tbd_0874). Both genes were cloned and expressed protein characterized. The larger protein (Tbd_0210; 544 residues) possesses an N-terminal ATP sulfurylase domain and a C-terminal APS kinase domain and was therefore annotated as a bifunctional enzyme. But, the protein was not bifunctional because it lacked ATP sulfurylase activity. However, the enzyme did possess APS kinase activity and displayed substrate inhibition by APS. Truncated protein missing the N-terminal domain had <2% APS kinase activity suggesting the function of the inactive sulfurylase domain is to promote the oligomerization of the APS kinase domains. The smaller gene product (Tbd_0874; 402 residues) possessed strong ATP sulfurylase activity with kinetic properties that appear to be kinetically optimized for the direction of APS utilization and ATP+sulfate production, which is consistent with an enzyme that functions physiologically to produce inorganic sulfate.

  20. Purification and properties of a dissimilatory nitrate reductase from Haloferax denitrificans

    NASA Technical Reports Server (NTRS)

    Hochstein, L. I.; Lang, F.

    1991-01-01

    A membrane-bound nitrate reductase (nitrite:(acceptor) oxidoreductase, EC 1.7.99.4) from the extremely halophilic bacterium Haloferax denitrificans was solubilized by incubating membranes in buffer lacking NaCl and purified by DEAE, hydroxylapatite, and Sepharose 6B gel filtration chromatography. The purified nitrate reductase reduced chlorate and was inhibited by azide and cyanide. Preincubating the enzyme with cyanide increased the extent of inhibition which in turn was intensified when dithionite was present. Although cyanide was a noncompetitive inhibitor with respect to nitrate, nitrate protected against inhibition. The enzyme, as isolated, was composed of two subunits (Mr 116,000 and 60,000) and behaved as a dimer during gel filtration (Mr 380,000). Unlike other halobacterial enzymes, this nitrate reductase was most active, as well as stable, in the absence of salt.

  1. Comparative Analysis of Denitrifying Activities of Hyphomicrobium nitrativorans, Hyphomicrobium denitrificans, and Hyphomicrobium zavarzinii.

    PubMed

    Martineau, Christine; Mauffrey, Florian; Villemur, Richard

    2015-08-01

    Hyphomicrobium spp. are commonly identified as major players in denitrification systems supplied with methanol as a carbon source. However, denitrifying Hyphomicrobium species are poorly characterized, and very few studies have provided information on the genetic and physiological aspects of denitrification in pure cultures of these bacteria. This is a comparative study of three denitrifying Hyphomicrobium species, H. denitrificans ATCC 51888, H. zavarzinii ZV622, and a newly described species, H. nitrativorans NL23, which was isolated from a denitrification system treating seawater. Whole-genome sequence analyses revealed that although they share numerous orthologous genes, these three species differ greatly in their nitrate reductases, with gene clusters encoding a periplasmic nitrate reductase (Nap) in H. nitrativorans, a membrane-bound nitrate reductase (Nar) in H. denitrificans, and one Nap and two Nar enzymes in H. zavarzinii. Concurrently with these differences observed at the genetic level, important differences in the denitrification capacities of these Hyphomicrobium species were determined. H. nitrativorans grew and denitrified at higher nitrate and NaCl concentrations than did the two other species, without significant nitrite accumulation. Significant increases in the relative gene expression levels of the nitrate (napA) and nitrite (nirK) reductase genes were also noted for H. nitrativorans at higher nitrate and NaCl concentrations. Oxygen was also found to be a strong regulator of denitrification gene expression in both H. nitrativorans and H. zavarzinii, although individual genes responded differently in these two species. Taken together, the results presented in this study highlight the potential of H. nitrativorans as an efficient and adaptable bacterium that is able to perform complete denitrification under various conditions. PMID:25979892

  2. Comparative Analysis of Denitrifying Activities of Hyphomicrobium nitrativorans, Hyphomicrobium denitrificans, and Hyphomicrobium zavarzinii

    PubMed Central

    Martineau, Christine; Mauffrey, Florian

    2015-01-01

    Hyphomicrobium spp. are commonly identified as major players in denitrification systems supplied with methanol as a carbon source. However, denitrifying Hyphomicrobium species are poorly characterized, and very few studies have provided information on the genetic and physiological aspects of denitrification in pure cultures of these bacteria. This is a comparative study of three denitrifying Hyphomicrobium species, H. denitrificans ATCC 51888, H. zavarzinii ZV622, and a newly described species, H. nitrativorans NL23, which was isolated from a denitrification system treating seawater. Whole-genome sequence analyses revealed that although they share numerous orthologous genes, these three species differ greatly in their nitrate reductases, with gene clusters encoding a periplasmic nitrate reductase (Nap) in H. nitrativorans, a membrane-bound nitrate reductase (Nar) in H. denitrificans, and one Nap and two Nar enzymes in H. zavarzinii. Concurrently with these differences observed at the genetic level, important differences in the denitrification capacities of these Hyphomicrobium species were determined. H. nitrativorans grew and denitrified at higher nitrate and NaCl concentrations than did the two other species, without significant nitrite accumulation. Significant increases in the relative gene expression levels of the nitrate (napA) and nitrite (nirK) reductase genes were also noted for H. nitrativorans at higher nitrate and NaCl concentrations. Oxygen was also found to be a strong regulator of denitrification gene expression in both H. nitrativorans and H. zavarzinii, although individual genes responded differently in these two species. Taken together, the results presented in this study highlight the potential of H. nitrativorans as an efficient and adaptable bacterium that is able to perform complete denitrification under various conditions. PMID:25979892

  3. Biosynthesis of the corrin macrocycle of coenzyme B12 in Pseudomonas denitrificans.

    PubMed Central

    Debussche, L; Thibaut, D; Cameron, B; Crouzet, J; Blanche, F

    1993-01-01

    Studies with cell-free protein preparations from a series of recombinant strains of Pseudomonas denitrificans demonstrated that precorrin-3 is converted into a further trimethylated intermediate, named precorrin-3B, along the pathway to coenzyme B12. It was then shown that the part of the pathway from precorrin-3 (called precorrin-3A hereafter) to precorrin-6x involves three intermediates, precorrin-3B, precorrin-4, and precorrin-5. Precorrin-3B was isolated in its native (reduced) as well as its oxidized (factor-IIIB) states, and precorrin-4 was isolated in its oxidized form only (factor-IV). Both factors were in vitro precursors of precorrin-6x. The synthesis of precorrin-6x from precorrin-3A was shown to be catalyzed by four enzymes, CobG, CobJ, CobM, and CobF, intervening in this order. They were purified to homogeneity. CobG, which converts precorrin-3A to precorrin-3B, was found to be an iron-sulfur protein responsible for the oxidation known to occur between precorrin-3A and precorrin-6x, and CobJ, CobM, and CobF are the C-17, C-11, and C-1 methylases, respectively. The acetate fragment is extruded after precorrin-4 formation. This study combined with our recent structural studies on factor-IV (D. Thibaut, L. Debussche, D. Fréchet, F. Herman, M. Vuilhorgne, and F. Blanche, J. Chem. Soc. Chem. Commun. 1993:513-515, 1993) and precorrin-3B (L. Debussche, D. Thibaut, M. Danzer, F. Debu, D. Fréchet, F. Herman, F. Blanche, and M. Vuilhorgne, J. Chem. Soc. Chem. Commun. 1993:1100-1103, 1993) provides a first step-by-step picture of the sequence of the enzymatic reactions leading to the corrin ring in P. denitrificans. PMID:8226690

  4. Expression studies on the ba3 quinol oxidase from Paracoccus denitrificans. A bb3 variant is enzymatically inactive.

    PubMed

    Zickermann, I; Tautu, O S; Link, T A; Korn, M; Ludwig, B; Richter, O M

    1997-06-15

    Expression of the quinol oxidase from Paracoccus denitrificans has been examined using a polyclonal antibody directed against subunit II and a promoter probe vector carrying the promoter region of the qox operon. Under aerobic conditions nitrate and nitrite act as specific inducers of the expression. To obtain an enzymatically competent quinol oxidase complex, an intact ctaB gene is required, which constitutes part of the cta operon coding for the aa3 cytochrome c oxidase of P. denitrificans. Deletion of ctaB leads to a change in heme composition of the quinol oxidase with heme b replacing the high-spin heme a of the binuclear center, causing loss of electron transport activity. PMID:9219517

  5. Anaerobic, Nitrate-Dependent Oxidation of U(IV) Oxide Minerals by the Chemolithoautotrophic Bacterium Thiobacillus denitrificans

    SciTech Connect

    Beller, H R

    2004-06-25

    Under anaerobic conditions and at circumneutral pH, cells of the widely-distributed, obligate chemolithoautotrophic bacterium Thiobacillus denitrificans oxidatively dissolved synthetic and biogenic U(IV) oxides (uraninite) in nitrate-dependent fashion: U(IV) oxidation required the presence of nitrate and was strongly correlated to nitrate consumption. This is the first report of anaerobic U(IV) oxidation by an autotrophic bacterium.

  6. Biosynthesis of medium chain length poly(3-hydroxyalkanoates) (mcl-PHAs) by Comamonas testosteroni during cultivation on vegetable oils.

    PubMed

    Thakor, Nehal; Trivedi, Ujjval; Patel, K C

    2005-11-01

    Comamonas testosteroni has been studied for its ability to synthesize and accumulate medium chain length poly(3-hydroxyalkanoates) (mcl-PHAs) during cultivation on vegetable oils available in the local market. Castor seed oil, coconut oil, mustard oil, cotton seed oil, groundnut oil, olive oil and sesame oil were supplemented in the mineral medium as a sole source of carbon for growth and PHAs accumulation. The composition of PHAs was analysed by a coupled gas chromatography/mass spectroscopy (GC/MS). PHAs contained C6 to C14 3-hydroxy acids, with a strong presence of 3-hydroxyoctanoate when coconut oil, mustard oil, cotton seed oil and groundnut oil were supplied. 3-hydroxydecanoate was incorporated at higher concentrations when castor seed oil, olive oil and sesame oil were the substrates. Purified PHAs samples were characterized by Fourier Transform Infrared (FTIR) and 13C NMR analysis. During cultivation on various vegetable oils, C. testosteroni accumulated PHAs up to 78.5-87.5% of the cellular dry material (CDM). The efficiency of the culture to convert oil to PHAs ranged from 53.1% to 58.3% for different vegetable oils. Further more, the composition of the PHAs formed was not found to be substrate dependent as PHAs obtained from C. testosteroni during growth on variety of vegetable oils showed similar compositions; 3-hydroxyoctanoic acid and/or 3-hydroxydecanoic acid being always predominant. The polymerizing system of C. testosteroni showed higher preference for C8 and C10 monomers as longer and smaller monomers were incorporated less efficiently. PMID:16084364

  7. Selenite reduction by the obligate aerobic bacterium Comamonas testosteroni S44 isolated from a metal-contaminated soil

    PubMed Central

    2014-01-01

    Background Selenium (Se) is an essential trace element in most organisms but has to be carefully handled since there is a thin line between beneficial and toxic concentrations. Many bacteria have the ability to reduce selenite (Se(IV)) and (or) selenate (Se(VI)) to red elemental selenium that is less toxic. Results A strictly aerobic bacterium, Comamonas testosteroni S44, previously isolated from metal(loid)-contaminated soil in southern China, reduced Se(IV) to red selenium nanoparticles (SeNPs) with sizes ranging from 100 to 200 nm. Both energy dispersive X-ray Spectroscopy (EDX or EDS) and EDS Elemental Mapping showed no element Se and SeNPs were produced inside cells whereas Se(IV) was reduced to red-colored selenium in the cytoplasmic fraction in presence of NADPH. Tungstate inhibited Se(VI) but not Se(IV) reduction, indicating the Se(IV)-reducing determinant does not contain molybdenum as co-factor. Strain S44 was resistant to multiple heavy and transition metal(loid)s such as Se(IV), As(III), Cu(II), and Cd(II) with minimal inhibitory concentrations (MIC) of 100 mM, 20 mM, 4 mM, and 0.5 mM, respectively. Disruption of iscR encoding a transcriptional regulator negatively impacted cellular growth and subsequent resistance to multiple heavy metal(loid)s. Conclusions C. testosteroni S44 could be very useful for bioremediation in heavy metal(loid) polluted soils due to the ability to both reduce toxic Se(VI) and Se(IV) to non-toxic Se (0) under aerobic conditions and to tolerate multiple heavy and transition metals. IscR appears to be an activator to regulate genes involved in resistance to heavy or transition metal(loid)s but not for genes responsible for Se(IV) reduction. PMID:25098921

  8. Biosynthesis of medium chain length poly(3-hydroxyalkanoates) (mcl-PHAs) by Comamonas testosteroni during cultivation on vegetable oils.

    PubMed

    Thakor, Nehal; Trivedi, Ujjval; Patel, K C

    2005-11-01

    Comamonas testosteroni has been studied for its ability to synthesize and accumulate medium chain length poly(3-hydroxyalkanoates) (mcl-PHAs) during cultivation on vegetable oils available in the local market. Castor seed oil, coconut oil, mustard oil, cotton seed oil, groundnut oil, olive oil and sesame oil were supplemented in the mineral medium as a sole source of carbon for growth and PHAs accumulation. The composition of PHAs was analysed by a coupled gas chromatography/mass spectroscopy (GC/MS). PHAs contained C6 to C14 3-hydroxy acids, with a strong presence of 3-hydroxyoctanoate when coconut oil, mustard oil, cotton seed oil and groundnut oil were supplied. 3-hydroxydecanoate was incorporated at higher concentrations when castor seed oil, olive oil and sesame oil were the substrates. Purified PHAs samples were characterized by Fourier Transform Infrared (FTIR) and 13C NMR analysis. During cultivation on various vegetable oils, C. testosteroni accumulated PHAs up to 78.5-87.5% of the cellular dry material (CDM). The efficiency of the culture to convert oil to PHAs ranged from 53.1% to 58.3% for different vegetable oils. Further more, the composition of the PHAs formed was not found to be substrate dependent as PHAs obtained from C. testosteroni during growth on variety of vegetable oils showed similar compositions; 3-hydroxyoctanoic acid and/or 3-hydroxydecanoic acid being always predominant. The polymerizing system of C. testosteroni showed higher preference for C8 and C10 monomers as longer and smaller monomers were incorporated less efficiently.

  9. Global Regulator IscR Positively Contributes to Antimonite Resistance and Oxidation in Comamonas testosteroni S44

    PubMed Central

    Liu, Hongliang; Zhuang, Weiping; Zhang, Shengzhe; Rensing, Christopher; Huang, Jun; Li, Jie; Wang, Gejiao

    2015-01-01

    Antimonial compounds can be found as a toxic contaminant in the environment. Knowledge on mechanisms of microbial Sb oxidation and its role in microbial tolerance are limited. Previously, we found that Comamonas testosteroni S44 was resistant to multiple heavy metals and was able to oxidize the toxic antimonite [Sb(III)] to the much less toxic antimonate [Sb(V)]. In this study, transposon mutagenesis was performed in C. testosteroni S44 to isolate genes responsible for Sb(III) resistance and oxidation. An insertion mutation into iscR, which regulates genes involved in the biosynthesis of Fe-S clusters, generated a strain called iscR-280. This mutant strain was complemented with a plasmid carrying iscR to generate strain iscR-280C. Compared to the wild type S44 and iscR-280C, strain iscR-280 showed lower resistance to Sb(III) and a lower Sb(III) oxidation rate. Strain iscR-280 also showed lower resistance to As(III), Cd(II), Cu(II), and H2O2. In addition, intracellular γ-glutamylcysteine ligase (γ-GCL) activity and glutathione (GSH) content were decreased in the mutated strain iscR-280. Real-time RT-PCR and lacZ fusion expression assay indicated that transcription of iscR and iscS was induced by Sb(III). Results of electrophoretic mobility shift assay (EMSA) and bacterial one-hybrid (B1H) system demonstrated a positive interaction between IscR and its promoter region. The diverse defective phenotypes and various expression patterns suggest a role for IscR in contributing to multi-metal(loid)s resistance and Sb(III) oxidation via Fe-S cluster biogenesis and oxidative stress protection. Bacterial Sb(III) oxidation is a detoxification reaction. PMID:26734615

  10. Investigating the Proton Donor in the NO Reductase from Paracoccus denitrificans

    PubMed Central

    ter Beek, Josy; Krause, Nils; Ädelroth, Pia

    2016-01-01

    Variant nomenclature: the variants were made in the NorB subunit if not indicated by the superscript c, which are variants in the NorC subunit (e.g. E122A = exchange of Glu-122 in NorB for an Ala, E71cD; exchange of Glu-71 in NorC for an Asp). Bacterial NO reductases (NORs) are integral membrane proteins from the heme-copper oxidase superfamily. Most heme-copper oxidases are proton-pumping enzymes that reduce O2 as the last step in the respiratory chain. With electrons from cytochrome c, NO reductase (cNOR) from Paracoccus (P.) denitrificans reduces NO to N2O via the following reaction: 2NO+2e-+2H+→N2O+H2O. Although this reaction is as exergonic as O2-reduction, cNOR does not contribute to the electrochemical gradient over the membrane. This means that cNOR does not pump protons and that the protons needed for the reaction are taken from the periplasmic side of the membrane (since the electrons are donated from this side). We previously showed that the P. denitrificans cNOR uses a single defined proton pathway with residues Glu-58 and Lys-54 from the NorC subunit at the entrance. Here we further strengthened the evidence in support of this pathway. Our further aim was to define the continuation of the pathway and the immediate proton donor for the active site. To this end, we investigated the region around the calcium-binding site and both propionates of heme b3 by site directed mutagenesis. Changing single amino acids in these areas often had severe effects on cNOR function, with many variants having a perturbed active site, making detailed analysis of proton transfer properties difficult. Our data does however indicate that the calcium ligation sphere and the region around the heme b3 propionates are important for proton transfer and presumably contain the proton donor. The possible evolutionary link between the area for the immediate donor in cNOR and the proton loading site (PLS) for pumped protons in oxygen-reducing heme-copper oxidases is discussed. PMID

  11. Mutational analysis of the nor gene cluster which encodes nitric-oxide reductase from Paracoccus denitrificans.

    PubMed

    de Boer, A P; van der Oost, J; Reijnders, W N; Westerhoff, H V; Stouthamer, A H; van Spanning, R J

    1996-12-15

    The genes that encode the hc-type nitric-oxide reductase from Paracoccus denitrificans have been identified. They are part of a cluster of six genes (norCBQDEF) and are found near the gene cluster that encodes the cd1-type nitrite reductase, which was identified earlier [de Boer, A. P. N., Reijnders, W. N. M., Kuenen, J. G., Stouthamer, A. H. & van Spanning, R. J. M. (1994) Isolation, sequencing and mutational analysis of a gene cluster involved in nitrite reduction in Paracoccus denitrificans, Antonie Leeu wenhoek 66, 111-127]. norC and norB encode the cytochrome-c-containing subunit II and cytochrome b-containing subunit I of nitric-oxide reductase (NO reductase), respectively. norQ encodes a protein with an ATP-binding motif and has high similarity to NirQ from Pseudomonas stutzeri and Pseudomonas aeruginosa and CbbQ from Pseudomonas hydrogenothermophila. norE encodes a protein with five putative transmembrane alpha-helices and has similarity to CoxIII, the third subunit of the aa3-type cytochrome-c oxidases. norF encodes a small protein with two putative transmembrane alpha-helices. Mutagenesis of norC, norB, norQ and norD resulted in cells unable to grow anaerobically. Nitrite reductase and NO reductase (with succinate or ascorbate as substrates) and nitrous oxide reductase (with succinate as substrate) activities were not detected in these mutant strains. Nitrite extrusion was detected in the medium, indicating that nitrate reductase was active. The norQ and norD mutant strains retained about 16% and 23% of the wild-type level of NorC, respectively. The norE and norF mutant strains had specific growth rates and NorC contents similar to those of the wild-type strain, but had reduced NOR and NIR activities, indicating that their gene products are involved in regulation of enzyme activity. Mutant strains containing the norCBQDEF region on the broad-host-range vector pEG400 were able to grow anaerobically, although at a lower specific growth rate and with lower

  12. Investigating the Proton Donor in the NO Reductase from Paracoccus denitrificans.

    PubMed

    ter Beek, Josy; Krause, Nils; Ädelroth, Pia

    2016-01-01

    Variant nomenclature: the variants were made in the NorB subunit if not indicated by the superscript c, which are variants in the NorC subunit (e.g. E122A = exchange of Glu-122 in NorB for an Ala, E71cD; exchange of Glu-71 in NorC for an Asp). Bacterial NO reductases (NORs) are integral membrane proteins from the heme-copper oxidase superfamily. Most heme-copper oxidases are proton-pumping enzymes that reduce O2 as the last step in the respiratory chain. With electrons from cytochrome c, NO reductase (cNOR) from Paracoccus (P.) denitrificans reduces NO to N2O via the following reaction: 2NO+2e-+2H+→N2O+H2O. Although this reaction is as exergonic as O2-reduction, cNOR does not contribute to the electrochemical gradient over the membrane. This means that cNOR does not pump protons and that the protons needed for the reaction are taken from the periplasmic side of the membrane (since the electrons are donated from this side). We previously showed that the P. denitrificans cNOR uses a single defined proton pathway with residues Glu-58 and Lys-54 from the NorC subunit at the entrance. Here we further strengthened the evidence in support of this pathway. Our further aim was to define the continuation of the pathway and the immediate proton donor for the active site. To this end, we investigated the region around the calcium-binding site and both propionates of heme b3 by site directed mutagenesis. Changing single amino acids in these areas often had severe effects on cNOR function, with many variants having a perturbed active site, making detailed analysis of proton transfer properties difficult. Our data does however indicate that the calcium ligation sphere and the region around the heme b3 propionates are important for proton transfer and presumably contain the proton donor. The possible evolutionary link between the area for the immediate donor in cNOR and the proton loading site (PLS) for pumped protons in oxygen-reducing heme-copper oxidases is discussed.

  13. Peroxidase Activity and Involvement in the Oxidative Stress Response of Roseobacter denitrificans Truncated Hemoglobin

    PubMed Central

    Wang, Yaya; Barbeau, Xavier; Bilimoria, Astha; Lagüe, Patrick; Couture, Manon; Tang, Joseph Kuo-Hsiang

    2015-01-01

    Roseobacter denitrificans is a member of the widespread marine Roseobacter genus. We report the first characterization of a truncated hemoglobin from R. denitrificans (Rd. trHb) that was purified in the heme-bound form from heterologous expression of the protein in Escherichia coli. Rd. trHb exhibits predominantly alpha-helical secondary structure and absorbs light at 412, 538 and 572 nm. The phylogenetic classification suggests that Rd. trHb falls into group II trHbs, whereas sequence alignments indicate that it shares certain important heme pocket residues with group I trHbs in addition to those of group II trHbs. The resonance Raman spectra indicate that the isolated Rd. trHb contains a ferric heme that is mostly 6-coordinate low-spin and that the heme of the ferrous form displays a mixture of 5- and 6-coordinate states. Two Fe-His stretching modes were detected, notably one at 248 cm-1, which has been reported in peroxidases and some flavohemoglobins that contain an Fe-His-Asp (or Glu) catalytic triad, but was never reported before in a trHb. We show that Rd. trHb exhibits a significant peroxidase activity with a (kcat/Km) value three orders of magnitude higher than that of bovine Hb and only one order lower than that of horseradish peroxidase. This enzymatic activity is pH-dependent with a pKa value ~6.8. Homology modeling suggests that residues known to be important for interactions with heme-bound ligands in group II trHbs from Mycobacterium tuberculosis and Bacillus subtilis are pointing toward to heme in Rd. trHb. Genomic organization and gene expression profiles imply possible functions for detoxification of reactive oxygen and nitrogen species in vivo. Altogether, Rd. trHb exhibits some distinctive features and appears equipped to help the bacterium to cope with reactive oxygen/nitrogen species and/or to operate redox biochemistry. PMID:25658318

  14. Phospholipid fatty acids as physiological indicators of Paracoccus denitrificans encapsulated in silica sol-gel hydrogels.

    PubMed

    Trögl, Josef; Jirková, Ivana; Kuráň, Pavel; Akhmetshina, Elmira; Brovdyová, Taťjána; Sirotkin, Alexander; Kirilina, Tatiana

    2015-01-01

    The phospholipid fatty acid (PLFA) content was determined in samples of Paracoccus denitrificans encapsulated in silica hydrogel films prepared from prepolymerized tetramethoxysilane (TMOS). Immediately after encapsulation the total PLFA concentration was linearly proportional to the optical density (600 nm) of the input microbial suspension (R2 = 0.99). After 7 days this relationship remained linear, but with significantly decreased slope, indicating a higher extinction of bacteria in suspensions of input concentration 108 cells/mL and higher. trans-Fatty acids, indicators of cytoplasmatic membrane disturbances, were below the detection limit. The cy/pre ratio (i.e., ratio of cyclopropylated fatty acids (cy17:0 + cy19:0) to their metabolic precursors (16:1ω7 + 18:1ω7)), an indicator of the transition of the culture to a stationary growth-phase, decreased depending on co-immobilization of nutrients in the order phosphate buffer > mineral medium > Luria Broth rich medium. The ratio, too, was logarithmically proportional to cell concentration. These results confirm the applicability of total PLFA as an indicator for the determination of living biomass and cy/pre ratio for determination of nutrient limitation of microorganisms encapsulated in sol-gel matrices. This may be of interest for monitoring of sol-gel encapsulated bacteria proposed as optical recognition elements in biosensor construction, as well as other biotechnological applications. PMID:25690547

  15. Phospholipid Fatty Acids as Physiological Indicators of Paracoccus denitrificans Encapsulated in Silica Sol-Gel Hydrogels

    PubMed Central

    Trögl, Josef; Jirková, Ivana; Kuráň, Pavel; Akhmetshina, Elmira; Brovdyová, Tat′jána; Sirotkin, Alexander; Kirilina, Tatiana

    2015-01-01

    The phospholipid fatty acid (PLFA) content was determined in samples of Paracoccus denitrificans encapsulated in silica hydrogel films prepared from prepolymerized tetramethoxysilane (TMOS). Immediately after encapsulation the total PLFA concentration was linearly proportional to the optical density (600 nm) of the input microbial suspension (R2 = 0.99). After 7 days this relationship remained linear, but with significantly decreased slope, indicating a higher extinction of bacteria in suspensions of input concentration 108 cells/mL and higher. trans-Fatty acids, indicators of cytoplasmatic membrane disturbances, were below the detection limit. The cy/pre ratio (i.e., ratio of cyclopropylated fatty acids (cy17:0 + cy19:0) to their metabolic precursors (16:1ω7 + 18:1ω7)), an indicator of the transition of the culture to a stationary growth-phase, decreased depending on co-immobilization of nutrients in the order phosphate buffer > mineral medium > Luria Broth rich medium. The ratio, too, was logarithmically proportional to cell concentration. These results confirm the applicability of total PLFA as an indicator for the determination of living biomass and cy/pre ratio for determination of nutrient limitation of microorganisms encapsulated in sol-gel matrices. This may be of interest for monitoring of sol-gel encapsulated bacteria proposed as optical recognition elements in biosensor construction, as well as other biotechnological applications. PMID:25690547

  16. Genome analysis and physiological comparison of Alicycliphilus denitrificans strains BC and K601T

    SciTech Connect

    Oosterkamp, Margreet J.; Veuskens, Teun; Saia, Flavia Talarico; Weelink, Sander A.B.; Goodwin, Lynne A.; Daligault, Hajnalka E.; Bruce, David; Detter, J. Chris; Tapia, Roxanne; Han, Cliff; Land, Miriam L; Hauser, Loren John; Langenhoff, A. M.; Gerritse, Jan; Van Berkel, Willem J. H.; Pieper, Dietmar; Junca, Howard; Smidt, Hauke; Schraa, Gosse; Davids, Mark; Schaap, Peter J; Plugge, Caroline M.; Stams, Alfons J. M.

    2013-01-01

    The genomes of the Betaproteobacteria Alicycliphilus denitrificans strains BC and K601T have been sequenced to get insight into the physiology of the two strains. Strain BC degrades benzene with chlorate as electron acceptor. The cyclohexanol-degrading denitrifying strain K601T is not able to use chlorate as electron acceptor, while strain BC cannot degrade cyclohexanol. The 16S rRNA sequences of strains BC and K601T are identical and the fatty acid methyl ester patterns of the strains are similar. Basic Local Alignment Search Tool (BLAST) analysis of predicted open reading frames of both strains showed most hits with Acidovorax sp. JS42, a bacterium that degrades nitro-aromatics. The genomes include strain-specific plasmids (pAlide201 in strain K601T and pAlide01 and pAlide02 in strain BC). Key genes of chlorate reduction in strain BC were located on a 120 kb megaplasmid (pAlide01), which was absent in strain K601T. Genes involved in cyclohexanol degradation were only found in strain K601T. Benzene and toluene are degraded via oxygenase-mediated pathways in both strains. Genes involved in the meta-cleavage pathway of catechol are present in the genomes of both strains. Strain BC also contains all genes of the ortho-cleavage pathway. The large number of mono- and dioxygenase genes in the genomes suggests that the two strains have a broader substrate range than known thus far.

  17. Genome Analysis and Physiological Comparison of Alicycliphilus denitrificans Strains BC and K601T

    PubMed Central

    Talarico Saia, Flávia; Weelink, Sander A. B.; Goodwin, Lynne A.; Daligault, Hajnalka E.; Bruce, David C.; Detter, John C.; Tapia, Roxanne; Han, Cliff S.; Land, Miriam L.; Hauser, Loren J.; Langenhoff, Alette A. M.; Gerritse, Jan; van Berkel, Willem J. H.; Pieper, Dietmar H.; Junca, Howard; Smidt, Hauke; Schraa, Gosse; Davids, Mark; Schaap, Peter J.; Plugge, Caroline M.; Stams, Alfons J. M.

    2013-01-01

    The genomes of the Betaproteobacteria Alicycliphilus denitrificans strains BC and K601T have been sequenced to get insight into the physiology of the two strains. Strain BC degrades benzene with chlorate as electron acceptor. The cyclohexanol-degrading denitrifying strain K601T is not able to use chlorate as electron acceptor, while strain BC cannot degrade cyclohexanol. The 16S rRNA sequences of strains BC and K601T are identical and the fatty acid methyl ester patterns of the strains are similar. Basic Local Alignment Search Tool (BLAST) analysis of predicted open reading frames of both strains showed most hits with Acidovorax sp. JS42, a bacterium that degrades nitro-aromatics. The genomes include strain-specific plasmids (pAlide201 in strain K601T and pAlide01 and pAlide02 in strain BC). Key genes of chlorate reduction in strain BC were located on a 120 kb megaplasmid (pAlide01), which was absent in strain K601T. Genes involved in cyclohexanol degradation were only found in strain K601T. Benzene and toluene are degraded via oxygenase-mediated pathways in both strains. Genes involved in the meta-cleavage pathway of catechol are present in the genomes of both strains. Strain BC also contains all genes of the ortho-cleavage pathway. The large number of mono- and dioxygenase genes in the genomes suggests that the two strains have a broader substrate range than known thus far. PMID:23825601

  18. Characterization of the p-toluenesulfonate operon tsaMBCD and tsaR in Comamonas testosteroni T-2.

    PubMed Central

    Junker, F; Kiewitz, R; Cook, A M

    1997-01-01

    Comamonas testosteroni T-2 uses a standard, if seldom examined, attack on an aromatic compound and oxygenates the side chain of p-toluenesulfonate (TS) (or p-toluenecarboxylate) to p-sulfobenzoate (or terephthalate) prior to complete oxidation. The expression of the first three catabolic enzymes in the pathway, the TS methyl-monooxygenase system (comprising reductase B and oxygenase M; TsaMB), p-sulfobenzyl alcohol dehydrogenase (TsaC), and p-sulfobenzaldehyde dehydrogenase (TsaD), is coregulated as regulatory unit R1 (H. R. Schlafli Oppenberg, G. Chen, T. Leisinger, and A. M. Cook, Microbiology [Reading] 141:1891-1899, 1995). The components of the oxygenase system were repurified, and the N-terminal amino acid sequences were confirmed and extended. An internal sequence of TsaM was obtained, and the identity of the [2Fe-2S] Rieske center was confirmed by electron paramagnetic resonance spectroscopy. We purified both dehydrogenases (TsaC and TsaD) and determined their molecular weights and N-terminal amino acid sequences. Oligonucleotides derived from the partial sequences of TsaM were used to identify cloned DNA from strain T-2, and about 6 kb of contiguous cloned DNA was sequenced. Regulatory unit R1 was presumed to represent a four-gene operon (tsaMBCD) which was regulated by the LysR-type regulator, TsaR, encoded by a deduced one-gene transcriptional unit. The genes for the inducible TS transport system were not at this locus. The oxygenase system was confirmed to be a class IA mononuclear iron oxygenase, and class IA can now be seen to have two evolutionary groups, the monooxygenases and the dioxygenases, though the divergence is limited to the oxygenase components. The alcohol dehydrogenase TsaC was confirmed to belong to the short-chain, zinc-independent dehydrogenases, and the aldehyde dehydrogenase TsaD was found to resemble several other aldehyde dehydrogenases. The operon and its putative regulator are compared with units of the TOL plasmid. PMID:9006050

  19. Defluviimonas denitrificans gen. nov., sp. nov., and Pararhodobacter aggregans gen. nov., sp. nov., non-phototrophic Rhodobacteraceae from the biofilter of a marine aquaculture.

    PubMed

    Foesel, Bärbel U; Drake, Harold L; Schramm, Andreas

    2011-11-01

    Three Gram-negative bacterial strains were isolated from the biofilter of a recirculating marine aquaculture. They were non-pigmented rods, mesophiles, moderately halophilic, and showed chemo-organoheterotrophic growth on various sugars, fatty acids, and amino acids, with oxygen as electron acceptor; strains D9-3(T) and D11-58 were in addition able to denitrify. Phototrophic or fermentative growth could not be demonstrated. Phylogenetic analysis of the 16S rRNA gene sequences placed D9-3(T) and D11-58, and D1-19(T) on two distinct branches within the alpha-3 proteobacterial Rhodobacteraceae, affiliated with, but clearly separate from, the genera Rhodobacter, Rhodovulum, and Rhodobaca. Based on morphological, physiological, and 16S rRNA-based phylogenetic characteristics, the isolated strains are proposed as new species of two novel genera, Defluviimonas denitrificans gen. nov., sp. nov. (type strain D9-3(T)=DSM 18921(T)=ATCC BAA-1447(T); additional strain D11-58=DSM19039=ATCC BAA-1448) and Pararhodobacter aggregans gen. nov., sp. nov (type strain D1-19(T)=DSM 18938(T)=ATCC BAA-1446(T)).

  20. Capillary zone electrophoresis with field enhanced sample stacking as a tool for targeted metabolome analysis of adenine nucleotides and coenzymes in Paracoccus denitrificans.

    PubMed

    Musilová, Jindra; Sedlácek, Vojtech; Kucera, Igor; Glatz, Zdenek

    2009-07-01

    The main aim of this work was to demonstrate the applicability of capillary zone electrophoresis in combination with field enhanced sample stacking in targeted metabolome analyses of adenine nucleotides--AMP, ADP, ATP, coenzymes NAD(+), NADP(+) and their reduced forms in Paracoccus denitrificans. Sodium carbonate/hydrogencarbonate buffer (100 mM, pH 9.6) with the addition of beta-CD at a concentration of 10 mM was found to be an effective BGE for their separation within 20 min. Besides this, special attention was paid to the development of the procedure for the extraction of specific metabolites from the bacterium P. denitrificans. This procedure was not only optimised to achieve the highest metabolite yields but also to obtain a sample that was fully compatible with the online preconcetration strategy used. The developed methodology was finally applied in a study of the bacterium P. denitrificans at various stages of the active respiratory chain.

  1. Inhibition of denitrification activity but not of mRNA induction in Paracoccus denitrificans by nitrite at a suboptimal pH.

    PubMed

    Baumann, B; van der Meer, J R; Snozzi, M; Zehnder, A J

    1997-10-01

    The influence of pH on the denitrification activity of a continuous culture of Paracoccus denitrificans was studied in relation to the presence of nitrite. After a transition from aerobic to anaerobic conditions at the suboptimal pH of 6.8, P. denitrificans was not able to build up a functional denitrification pathway. Nitrite accumulated in the medium as the predominant denitrification product. Although the nitrite reductase gene was induced properly, the enzyme could not be detected at sufficient amounts in the culture. These observations was somehow inhibited, or once synthesized nitrite reductase was inactivated, possibly by the high concentrations of nitrous acid (HNO2). Interestingly, when a P. denitrificans culture which was grown to steady-state under anaerobic conditions was then exposed to suboptimal pHs, cells exhibited a reduced overall denitrification activity, but neither nitrite nor any other denitrification intermediate accumulated. PMID:9403103

  2. Genome-enabled studies of anaerobic, nitrate-dependent iron oxidation in the chemolithoautotrophic bacterium Thiobacillus denitrificans

    PubMed Central

    Beller, Harry R.; Zhou, Peng; Legler, Tina C.; Chakicherla, Anu; Kane, Staci; Letain, Tracy E.; A. O’Day, Peggy

    2013-01-01

    Thiobacillus denitrificans is a chemolithoautotrophic bacterium capable of anaerobic, nitrate-dependent U(IV) and Fe(II) oxidation, both of which can strongly influence the long-term efficacy of in situ reductive immobilization of uranium in contaminated aquifers. We previously identified two c-type cytochromes involved in nitrate-dependent U(IV) oxidation in T. denitrificans and hypothesized that c-type cytochromes would also catalyze Fe(II) oxidation, as they have been found to play this role in anaerobic phototrophic Fe(II)-oxidizing bacteria. Here we report on efforts to identify genes associated with nitrate-dependent Fe(II) oxidation, namely (a) whole-genome transcriptional studies [using FeCO3, Fe2+, and U(IV) oxides as electron donors under denitrifying conditions], (b) Fe(II) oxidation assays performed with knockout mutants targeting primarily highly expressed or upregulated c-type cytochromes, and (c) random transposon-mutagenesis studies with screening for Fe(II) oxidation. Assays of mutants for 26 target genes, most of which were c-type cytochromes, indicated that none of the mutants tested were significantly defective in nitrate-dependent Fe(II) oxidation. The non-defective mutants included the c1-cytochrome subunit of the cytochrome bc1 complex (complex III), which has relevance to a previously proposed role for this complex in nitrate-dependent Fe(II) oxidation and to current concepts of reverse electron transfer. A transposon mutant with a disrupted gene associated with NADH:ubiquinone oxidoreductase (complex I) was ~35% defective relative to the wild-type strain; this strain was similarly defective in nitrate reduction with thiosulfate as the electron donor. Overall, our results indicate that nitrate-dependent Fe(II) oxidation in T. denitrificans is not catalyzed by the same c-type cytochromes involved in U(IV) oxidation, nor have other c-type cytochromes yet been implicated in the process. PMID:24065960

  3. Construction of a Specialized Cloning Strain of E. Coli for the Nitrate Reductase Genes of Haloferax Denitrificans

    NASA Technical Reports Server (NTRS)

    Johnson, Emmett

    1999-01-01

    This is the final report on Joint Research Interchange (NCC2-5011) "Construction of a Specialized Cloning Strain of E.. coli for the Nitrate Reductase Genes of Haloferax denitrificans." Originally the award was 11/l/93-10/31/95, but there were no-cost extensions made, because of a year Sabbatical at the Pasteur Institute in Paris and other leaves of 3 months each at the Pasteur Institute, during which work could not be done on this project, which extended the closing date to 10/30/98.

  4. Biodegradation of naphthalene-2-sulfonic acid present in tannery wastewater by bacterial isolates Arthrobacter sp. 2AC and Comamonas sp. 4BC.

    PubMed

    Song, Zhi; Edwards, Suzanne R; Burns, Richard G

    2005-06-01

    Two bacterial strains, 2AC and 4BC, both capable of utilizing naphthalene-2-sulfonic acid (2-NSA) as a sole source of carbon, were isolated from activated sludges previously exposed to tannery wastewater. Enrichments were carried out in mineral salt medium (MSM) with 2-NSA as the sole carbon source. 16S rDNA sequencing analysis indicated that 2AC is an Arthrobacter sp. and 4BC is a Comamonas sp. Within 33 h, both isolates degraded 100% of 2-NSA in MSM and also 2-NSA in non-sterile tannery wastewater. The yield coefficient was 0.33 g biomass dry weight per gram of 2-NSA. A conceptual model, which describes the aerobic transformation of organic matter, was used for interpreting the biodegradation kinetics of 2-NSA. The half-lives for 2-NSA, at initial concentrations of 100 and 500 mg/l in MSM, ranged from 20 h (2AC) to 26 h (4BC) with lag-phases of 8 h (2AC) and 12 h (4BC). The carbon balance indicates that 75-90% of the initial TOC (total organic carbon) was mineralized, 5-20% remained as DOC (dissolved organic carbon) and 3-10% was biomass carbon. The principal metabolite of 2-NSA biodegradation (in both MSM and tannery wastewater) produced by Comamonas sp. 4BC had a MW of 174 and accounted for the residual DOC (7.0-19.0% of the initial TOC and 66% of the remaining TOC). Three to ten percent of the initial TOC (33% of the remaining TOC) was associated with biomass. The metabolite was not detected when Arthrobacter sp. 2AC was used, and a lower residual DOC and biomass carbon were recorded. This suggests that the two strains may use different catabolic pathways for 2-NSA degradation. The rapid biodegradation of 2-NSA (100 mg/l) added to non-sterile tannery wastewater (total 2-NSA, 105 mg/l) when inoculated with either Arthrobacter 2AC or Comamonas 4BC showed that both strains were able to compete with the indigenous microorganisms and degrade 2-NSA even in the presence of alternate carbon sources (DOC in tannery wastewater = 91 mg/l). The results provide information

  5. Burkholderia denitrificans sp. nov., isolated from the soil of Dokdo Island, Korea.

    PubMed

    Lee, Chang-Muk; Weon, Hang-Yeon; Yoon, Sang-Hong; Kim, Soo-Jin; Koo, Bon-Sung; Kwon, Soon-Wo

    2012-10-01

    A novel, Gram-negative, bacterial strain KIS30-44(T) was identified from wet forest soil collected on the Korean island of Dokdo. Growth of the strain was observed at 15-30°C, pH 5-9, 0-3% NaCl, and 950 mM KNO(3). KIS30-44(T) reduced nitrate to nitrogen gas. Analysis of the 16S rRNA gene sequence showed that KIS30-44(T) was phylogenetically related to Burkholderia sacchari, Burkholderia mimosarum, and Burkholderia oxyphila (98.1%, 98.0%, and 98.0% sequence similarity, respectively). The genomic G+C content was 63.5 mol%. KIS30-44(T) exhibited less than 52% DNA-DNA relatedness with the type strains of 9 closely related Burkholderia species. The major isoprenoid quinone was Q-8. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and two unknown aminolipids. The major fatty acids in KIS30-44(T) were C(16:0), C(18:1) ω7c and summed feature 3 (iso-C(15:0) 2-OH and C(16:1) ω7c), and the strain contained half the amount of C(17:0) cyclo found in the 9 closely related Burkholderia species. The results of these phenotypic, 16S rRNA gene sequence, DNA-DNA hybridization, and chemotaxonomic data indicate that KIS30-44(T) represents a novel species within the genus Burkholderia, for which the name Burkholderia denitrificans (Type strain KIS30-44(T) =KACC 12733(T) =DSM 24336(T)) is proposed.

  6. Biochemical properties of Paracoccus denitrificans FnrP: reactions with molecular oxygen and nitric oxide.

    PubMed

    Crack, Jason C; Hutchings, Matthew I; Thomson, Andrew J; Le Brun, Nick E

    2016-03-01

    In Paracoccus denitrificans, three CRP/FNR family regulatory proteins, NarR, NnrR and FnrP, control the switch between aerobic and anaerobic (denitrification) respiration. FnrP is a [4Fe-4S] cluster-containing homologue of the archetypal O2 sensor FNR from E. coli and accordingly regulates genes encoding aerobic and anaerobic respiratory enzymes in response to O2, and also NO, availability. Here we show that FnrP undergoes O2-driven [4Fe-4S] to [2Fe-2S] cluster conversion that involves up to 2 O2 per cluster, with significant oxidation of released cluster sulfide to sulfane observed at higher O2 concentrations. The rate of the cluster reaction was found to be ~sixfold lower than that of E. coli FNR, suggesting that FnrP can remain transcriptionally active under microaerobic conditions. This is consistent with a role for FnrP in activating expression of the high O2 affinity cytochrome c oxidase under microaerobic conditions. Cluster conversion resulted in dissociation of the transcriptionally active FnrP dimer into monomers. Therefore, along with E. coli FNR, FnrP belongs to the subset of FNR proteins in which cluster type is correlated with association state. Interestingly, two key charged residues, Arg140 and Asp154, that have been shown to play key roles in the monomer-dimer equilibrium in E. coli FNR are not conserved in FnrP, indicating that different protomer interactions are important for this equilibrium. Finally, the FnrP [4Fe-4S] cluster is shown to undergo reaction with multiple NO molecules, resulting in iron nitrosyl species and dissociation into monomers. PMID:26790880

  7. Iron-sulfur cluster-dependent catalysis of chlorophyllide a oxidoreductase from Roseobacter denitrificans.

    PubMed

    Kiesel, Svenja; Wätzlich, Denise; Lange, Christiane; Reijerse, Edward; Bröcker, Markus J; Rüdiger, Wolfhart; Lubitz, Wolfgang; Scheer, Hugo; Moser, Jürgen; Jahn, Dieter

    2015-01-01

    Bacteriochlorophyll a biosynthesis requires the stereo- and regiospecific two electron reduction of the C7-C8 double bond of chlorophyllide a by the nitrogenase-like multisubunit metalloenzyme, chlorophyllide a oxidoreductase (COR). ATP-dependent COR catalysis requires interaction of the protein subcomplex (BchX)2 with the catalytic (BchY/BchZ)2 protein to facilitate substrate reduction via two redox active iron-sulfur centers. The ternary COR enzyme holocomplex comprising subunits BchX, BchY, and BchZ from the purple bacterium Roseobacter denitrificans was trapped in the presence of the ATP transition state analog ADP·AlF4(-). Electron paramagnetic resonance experiments revealed a [4Fe-4S] cluster of subcomplex (BchX)2. A second [4Fe-4S] cluster was identified on (BchY/BchZ)2. Mutagenesis experiments indicated that the latter is ligated by four cysteines, which is in contrast to the three cysteine/one aspartate ligation pattern of the closely related dark-operative protochlorophyllide a oxidoreductase (DPOR). In subsequent mutagenesis experiments a DPOR-like aspartate ligation pattern was implemented for the catalytic [4Fe-4S] cluster of COR. Artificial cluster formation for this inactive COR variant was demonstrated spectroscopically. A series of chemically modified substrate molecules with altered substituents on the individual pyrrole rings and the isocyclic ring were tested as COR substrates. The COR enzyme was still able to reduce the B ring of substrates carrying modified substituents on ring systems A, C, and E. However, substrates with a modification of the distantly located propionate side chain were not accepted. A tentative substrate binding mode was concluded in analogy to the related DPOR system.

  8. Cytochrome cb-type nitric oxide reductase with cytochrome c oxidase activity from Paracoccus denitrificans ATCC 35512.

    PubMed

    Fujiwara, T; Fukumori, Y

    1996-04-01

    A highly active nitric oxide reductase was purified from Paracoccus denitrificans ATCC 35512, formerly named Thiosphaera pantotropha, which was anaerobically cultivated in the presence of nitrate. The enzyme was composed of two subunits with molecular masses of 34 and 15 kDa and contained two hemes b and one heme c per molecule. Copper was not found in the enzyme. The spectral properties suggested that one of the two hemes b and heme c were in six-coordinated low-spin states and another heme b was in a five-coordinated high-spin state and reacted with carbon monoxide. The enzyme showed high cytochrome c-nitric oxide oxidoreductase activity and formed nitrous oxide from nitric oxide with the expected stoichiometry when P. denitrificans ATCC 35512 ferrocytochrome c-550 was used as the electron donor. The V max and Km values for nitric oxide were 84 micromol of nitric oxide per min/mg of protein and 0.25 microM, respectively. Furthermore, the enzyme showed ferrocytochrome c-550-O2 oxidoreductase activity with a V max of 8.4 micromol of O2 per min/mg of protein and a Km value of 0.9 mM. Both activities were 50% inhibited by about 0.3 mM KCN. PMID:8606159

  9. Assembly of respiratory complexes I, III, and IV into NADH oxidase supercomplex stabilizes complex I in Paracoccus denitrificans.

    PubMed

    Stroh, Anke; Anderka, Oliver; Pfeiffer, Kathy; Yagi, Takao; Finel, Moshe; Ludwig, Bernd; Schägger, Hermann

    2004-02-01

    Stable supercomplexes of bacterial respiratory chain complexes III (ubiquinol:cytochrome c oxidoreductase) and IV (cytochrome c oxidase) have been isolated as early as 1985 (Berry, E. A., and Trumpower, B. L. (1985) J. Biol. Chem. 260, 2458-2467). However, these assemblies did not comprise complex I (NADH:ubiquinone oxidoreductase). Using the mild detergent digitonin for solubilization of Paracoccus denitrificans membranes we could isolate NADH oxidase, assembled from complexes I, III, and IV in a 1:4:4 stoichiometry. This is the first chromatographic isolation of a complete "respirasome." Inactivation of the gene for tightly bound cytochrome c552 did not prevent formation of this supercomplex, indicating that this electron carrier protein is not essential for structurally linking complexes III and IV. Complex I activity was also found in the membranes of mutant strains lacking complexes III or IV. However, no assembled complex I but only dissociated subunits were observed following the same protocols used for electrophoretic separation or chromatographic isolation of the supercomplex from the wild-type strain. This indicates that the P. denitrificans complex I is stabilized by assembly into the NADH oxidase supercomplex. In addition to substrate channeling, structural stabilization of a membrane protein complex thus appears as one of the major functions of respiratory chain supercomplexes.

  10. Enzymatic properties, evidence for in vivo expression, and intracellular localization of shewasin D, the pepsin homolog from Shewanella denitrificans

    PubMed Central

    Leal, Ana Rita; Cruz, Rui; Bur, Daniel; Huesgen, Pitter F.; Faro, Rosário; Manadas, Bruno; Wlodawer, Alexander; Faro, Carlos; Simões, Isaura

    2016-01-01

    The widespread presence of pepsin-like enzymes in eukaryotes together with their relevance in the control of multiple biological processes is reflected in the large number of studies published so far for this family of enzymes. By contrast, pepsin homologs from bacteria have only recently started to be characterized. The work with recombinant shewasin A from Shewanella amazonensis provided the first documentation of this activity in prokaryotes. Here we extend our studies to shewasin D, the pepsin homolog from Shewanella denitrificans, to gain further insight into this group of bacterial peptidases that likely represent ancestral versions of modern eukaryotic pepsin-like enzymes. We demonstrate that the enzymatic properties of recombinant shewasin D are strongly reminiscent of eukaryotic pepsin homologues. We determined the specificity preferences of both shewasin D and shewasin A using proteome-derived peptide libraries and observed remarkable similarities between both shewasins and eukaryotic pepsins, in particular with BACE-1, thereby confirming their phylogenetic proximity. Moreover, we provide first evidence of expression of active shewasin D in S. denitrificans cells, confirming its activity at acidic pH and inhibition by pepstatin. Finally, our results revealed an unprecedented localization for a family A1 member by demonstrating that native shewasin D accumulates preferentially in the cytoplasm. PMID:27029611

  11. Tightly bound nucleotides of the energy-transducing ATPase, and their role in oxidative phosphorylation. I. The Paracoccus denitrificans system.

    PubMed

    Harris, D A; John, P; Radda, G K

    1977-03-11

    1. The coupling ATPase of Paracoccus denitrificans can be removed from the membrane by washing coupled membrane fragments at low salt concentrations. 2. This ATPase resembles coupling ATPases of mitochondria, chloroplasts and other bacteria. It is a negatively charged protein of molecular weight about 300,000. An inhibitor protein in bound tightly to the ATPase in vivo, and can be destroyed by trypsin treatment. 3. ATP and ADP are found tightly bound to the coupling ATPase of P. denitrificans, both in its membrane-bound and isolated state. The ATP/ADP ratio on the enzyme is greater than one. 4. Under de-energised condtions, the bound nucleotides are not available to the suspending medium. When the membrane is energised however, the bound nucleotides can exchange with added nucleotides and incorporate 32Pi. 32Ppi is incorporated into the beta and gamma positions of the bound nucleotides, but beta-labelling probably does not occur on the coupling ATPase. 5. Uncouplers inhibit the exchange of the free nucleotides or 32Pi into the bound nucleotides, while venturicidin (an energy transfer inhibitor) and aurovertin stimulate the exchange. 6. The response of the bound nucleotides to energisation is consistent with their being involved directly in the mechanism of oxidative phosphorylation.

  12. Industrial vitamin B12 production by Pseudomonas denitrificans using maltose syrup and corn steep liquor as the cost-effective fermentation substrates.

    PubMed

    Xia, Wei; Chen, Wei; Peng, Wei-Fu; Li, Kun-Tai

    2015-06-01

    The aerobic Pseudomonas denitrificans is widely used for industrial and commercial vitamin B12 fermentation, due to its higher productivity compared to the anaerobic vitamin B12-producing microorganisms. This paper aimed to develop a cost-effective fermentation medium for industrial vitamin B12 production by P. denitrificans in 120,000-l fermenter. It was found that maltose syrup (a low-cost syrup from corn starch by means of enzymatic or acid hydrolysis) and corn steep liquor (CSL, a by-product of starch industry) were greatly applicable to vitamin B12 production by P. denitrificans. Under the optimal fermentation medium performed by response surface methodology, 198.27 ± 4.60 mg/l of vitamin B12 yield was obtained in 120,000-l fermenter, which was close to the fermentation with the refined sucrose (198.80 mg/l) and was obviously higher than that obtained under beet molasses utilization (181.75 mg/l). Therefore, maltose syrups and CSL were the efficient and economical substrates for industrial vitamin B12 fermentation by P. denitrificans. PMID:25561346

  13. Efficient simultaneous adsorption-biodegradation of high-concentrated N,N-dimethylformamide from water by Paracoccus denitrificans-graphene oxide microcomposites

    NASA Astrophysics Data System (ADS)

    Zheng, Yuan; Chen, Dongyun; Li, Najun; Xu, Qingfeng; Li, Hua; He, Jinghui; Lu, Jianmei

    2016-02-01

    Water contamination becomes one of the most pervasive environmental issues all over the world in recent years. In this paper, the functionalization of graphene oxide (GO) with copolymers containing methacrylic acid (MAA) and butyl methacrylate (BMA) was investigated to prepare a new microcomposite material (PGO) via free radical solution polymerization. PGO was used for the adsorption of N,N-dimethylformamide (DMF) from aqueous solution by utilizing the characteristics of ultralarge surface and the Van der Waals force between DMF molecules and polymers on the surface of PGO. Besides, PGO was used not only a high-capable adsorbent but also a carrier for the immobilization of Paracoccus denitrificans cells in the treatment of high-concentrated DMF. Bacterial cells could immobilized on the PGO (PGO@P. denitrificans) stably by covalent coupling process after acclimatization and high-concentrated DMF (2000 mg/L) could be removed completely and relatively rapidly from aqueous solutions by the simultaneous adsorption-biodegradation (SAB) process of PGO@P. denitrificans. Furthermore, the excellent recycle performance of PGO@P. denitrificans made the whole process more economical and practical.

  14. Efficient simultaneous adsorption-biodegradation of high-concentrated N,N-dimethylformamide from water by Paracoccus denitrificans-graphene oxide microcomposites

    PubMed Central

    Zheng, Yuan; Chen, Dongyun; Li, Najun; Xu, Qingfeng; Li, Hua; He, Jinghui; Lu, Jianmei

    2016-01-01

    Water contamination becomes one of the most pervasive environmental issues all over the world in recent years. In this paper, the functionalization of graphene oxide (GO) with copolymers containing methacrylic acid (MAA) and butyl methacrylate (BMA) was investigated to prepare a new microcomposite material (PGO) via free radical solution polymerization. PGO was used for the adsorption of N,N-dimethylformamide (DMF) from aqueous solution by utilizing the characteristics of ultralarge surface and the Van der Waals force between DMF molecules and polymers on the surface of PGO. Besides, PGO was used not only a high-capable adsorbent but also a carrier for the immobilization of Paracoccus denitrificans cells in the treatment of high-concentrated DMF. Bacterial cells could immobilized on the PGO (PGO@P. denitrificans) stably by covalent coupling process after acclimatization and high-concentrated DMF (2000 mg/L) could be removed completely and relatively rapidly from aqueous solutions by the simultaneous adsorption-biodegradation (SAB) process of PGO@P. denitrificans. Furthermore, the excellent recycle performance of PGO@P. denitrificans made the whole process more economical and practical. PMID:26829653

  15. Secondary metabolites extracted from marine sponge associated Comamonas testosteroni and Citrobacter freundii as potential antimicrobials against MDR pathogens and hypothetical leads for VP40 matrix protein of Ebola virus: an in vitro and in silico investigation.

    PubMed

    Skariyachan, Sinosh; Acharya, Archana B; Subramaniyan, Saumya; Babu, Sumangala; Kulkarni, Shruthi; Narayanappa, Rajeswari

    2016-09-01

    The current study explores therapeutic potential of metabolites extracted from marine sponge (Cliona sp.)-associated bacteria against MDR pathogens and predicts the binding prospective of probable lead molecules against VP40 target of Ebola virus. The metabolite-producing bacteria were characterized by agar overlay assay and as per the protocols in Bergey's manual of determinative bacteriology. The antibacterial activities of extracted metabolites were tested against clinical pathogens by well-diffusion assay. The selected metabolite producers were characterized by 16S rDNA sequencing. Chemical screening and Fourier Transform Infrared (FTIR) analysis for selected compounds were performed. The probable lead molecules present in the metabolites were hypothesized based on proximate analysis, FTIR data, and literature survey. The drug-like properties and binding potential of lead molecules against VP40 target of Ebola virus were hypothesized by computational virtual screening and molecular docking. The current study demonstrated that clear zones around bacterial colonies in agar overlay assay. Antibiotic sensitivity profiling demonstrated that the clinical isolates were multi-drug resistant, however; most of them showed sensitivity to secondary metabolites (MIC-15 μl/well). The proximate and FTIR analysis suggested that probable metabolites belonged to alkaloids with O-H, C-H, C=O, and N-H groups. 16S rDNA characterization of selected metabolite producers demonstrated that 96% and 99% sequence identity to Comamonas testosteroni and Citrobacter freundii, respectively. The docking studies suggested that molecules such as Gymnastatin, Sorbicillactone, Marizomib, and Daryamide can designed as probable lead candidates against VP40 target of Ebola virus.

  16. Structure of apo-azurin from Alcaligenes denitrificans at 1.8 A resolution.

    PubMed

    Shepard, W E; Kingston, R L; Anderson, B F; Baker, E N

    1993-05-01

    The structure of apo-azurin from Alcaligenes denitrificans has been determined at high resolution by X-ray crystallography. Two separate structure analyses have been carried out, (i) on crystals obtained from solutions of apo-azurin and (ii) on crystals obtained by removal of copper from previously formed crystals of holo-azurin. Data to 1.8 A resolution were collected from the apo-azurin crystals, by Weissenberg photography (with image plates) using synchrotron radiation and by diffractometry, and the structure was refined by restrained least-squares methods to a final R value of 0.160 for all data in the range 10.0-1.8 A. The final model of 1954 protein atoms, 246 water molecules (66 half-weighted), four SO(4)(2-) ions, and two low-occupancy (0.13 and 0.15) Cu atoms has r.m.s. deviations of 0.012, 0.045 and 0.013 A from standard bond lengths, angle distances and planar groups. For copper-removed azurin, data to 2.2 A were collected by diffractometry and the structure refined by restrained least squares to a final R value of 0.158 for all data in the range 10.0-2.2 A. The final model of 1954 protein atoms, 264 water molecules, two SO(4)(2-) ions, two low occupancy (0.18 and 0.22) metal atoms and one unidentified atom (modelled as S) has r.m.s. deviations of 0.013, 0.047 and 0.012 A from standard bond lengths, angle distances and planar groups. The two structures are essentially identical to each other and show no significant differences from the oxidized and reduced holo-azurin structures. The ligand side chains move slightly closer together following the removal of copper, with the radius of the cavity between the three strongly binding ligands, His 46, His 117 and Cys 112, shrinking from 1.31 A in reduced azurin to 1.24 A in oxidized azurin and 1.16 A in apo-azurin. There is a suggestion of increased flexibility in one of the copper-binding loops but the structure supports the view that the copper site found in holo-azurin is a stable structure, defined by the

  17. Genome-Enabled Studies of Anaerobic, Nitrate-Dependent Iron Oxidation in the Chemolithoautotrophic Bacterium Thiobacillus denitrificans

    NASA Astrophysics Data System (ADS)

    Beller, H. R.; Zhou, P.; Legler, T. C.; Chakicherla, A.; O'Day, P. A.

    2013-12-01

    Thiobacillus denitrificans is a chemolithoautotrophic bacterium capable of anaerobic, nitrate-dependent U(IV) and Fe(II) oxidation, both of which can strongly influence the long-term efficacy of in situ reductive immobilization of uranium in contaminated aquifers. We previously identified two c-type cytochromes involved in nitrate-dependent U(IV) oxidation in T. denitrificans and hypothesized that c-type cytochromes would also catalyze Fe(II) oxidation, as they have been found to play this role in anaerobic phototrophic Fe(II)-oxidizing bacteria. Here we report on efforts to identify genes associated with nitrate-dependent Fe(II) oxidation, namely (a) whole-genome transcriptional studies [using FeCO3, Fe2+, and U(IV) oxides as electron donors under denitrifying conditions], (b) Fe(II) oxidation assays performed with knockout mutants targeting primarily highly expressed or upregulated c-type cytochromes, and (c) random transposon-mutagenesis studies with screening for Fe(II) oxidation. Assays of mutants for 26 target genes, most of which were c-type cytochromes, indicated that none of the mutants tested were significantly defective in nitrate-dependent Fe(II) oxidation. The non-defective mutants included the c1-cytochrome subunit of the cytochrome bc1 complex (complex III), which has relevance to a previously proposed role for this complex in nitrate-dependent Fe(II) oxidation and to current concepts of reverse electron transfer. Of the transposon mutants defective in Fe(II) oxidation, one mutant with a disrupted gene associated with NADH:ubiquinone oxidoreductase (complex I) was ~35% defective relative to the wild-type strain; this strain was similarly defective in nitrate reduction with thiosulfate as the electron donor. Overall, our results indicate that nitrate-dependent Fe(II) oxidation in T. denitrificans is not catalyzed by the same c-type cytochromes involved in U(IV) oxidation, nor have other c-type cytochromes yet been implicated in the process.

  18. Anaerobic, Nitrate-Dependent Oxidation of Uraninite by the Chemolithoautotroph Thiobacillus denitrificans: Cell Suspension and Whole-Genome Transcriptional Studies

    NASA Astrophysics Data System (ADS)

    Beller, H. R.; Chakicherla, A.; Legler, T. C.; Letain, T. E.; Coleman, M.; Kane, S. R.

    2005-12-01

    Background: In-situ, reductive immobilization of uranium in aquifers, whereby relatively soluble U(VI) species are reduced to poorly soluble uraninite (UO2) by aquifer bacteria, has been the subject of intensive research effort recently. This study explored the possibility that a widespread soil bacterium, Thiobacillus denitrificans, could catalyze anaerobic U re-oxidation in the presence of nitrate, a common co-contaminant with uranium at U.S. DOE sites. Whole-genome, cDNA microarray studies (representing all 2832 ORFs of the 2.9 Mb genome) were conducted to identify genes upregulated during nitrate-dependent U(IV) oxidation (relative to control conditions of nitrate-dependent thiosulfate oxidation). Methods: Washed cell suspension experiments were carried out under strictly anaerobic conditions and at circumneutral pH with UO2 and T. denitrificans cells grown under denitrifying conditions and harvested in late exponential phase. Experiments included both sterile controls and live, no-nitrate controls. For microarray analysis, RNA was isolated from cells exposed to either UO2 or thiosulfate under strictly anaerobic, denitrifying conditions. For all samples analyzed with microarrays, chemical analyses were used to confirm that the applicable metabolic activity [i.e., denitrification and either U(IV) or thiosulfate oxidation] was occurring. Reverse transcription, quantitative PCR was used to confirm selected microarray results. Results: In the cell suspension experiments, T. denitrificans cells oxidatively dissolved UO2 in nitrate-dependent fashion: U(IV) oxidation required the presence of nitrate ( P<0.01) and was strongly correlated to nitrate consumption (r2 = 0.98). However, U(IV) oxidation and denitrification appeared to be dependent on H2. The microarrays identified 333 genes as upregulated under U(IV)-oxidizing conditions using RMA statistical analysis and a 2-fold ( P<0.0001) cutoff. Notably, 16 of these genes, which were upregulated 5- to 22-fold, were

  19. Electrochemistry suggests proton access from the exit site to the binuclear center in Paracoccus denitrificans cytochrome c oxidase pathway variants.

    PubMed

    Meyer, Thomas; Melin, Frédéric; Richter, Oliver-M H; Ludwig, Bernd; Kannt, Aimo; Müller, Hanne; Michel, Hartmut; Hellwig, Petra

    2015-02-27

    Two different pathways through which protons access cytochrome c oxidase operate during oxygen reduction from the mitochondrial matrix, or the bacterial cytoplasm. Here, we use electrocatalytic current measurements to follow oxygen reduction coupled to proton uptake in cytochrome c oxidase isolated from Paracoccus denitrificans. Wild type enzyme and site-specific variants with defects in both proton uptake pathways (K354M, D124N and K354M/D124N) were immobilized on gold nanoparticles, and oxygen reduction was probed electrochemically in the presence of varying concentrations of Zn(2+) ions, which are known to inhibit both the entry and the exit proton pathways in the enzyme. Our data suggest that under these conditions substrate protons gain access to the oxygen reduction site via the exit pathway.

  20. Precorrin-6x reductase from Pseudomonas denitrificans: purification and characterization of the enzyme and identification of the structural gene.

    PubMed Central

    Blanche, F; Thibaut, D; Famechon, A; Debussche, L; Cameron, B; Crouzet, J

    1992-01-01

    Precorrin-6x reductase, which catalyzes the NADPH-dependent reduction of precorrin-6x to a dihydro derivative named precorrin-6y, was purified 14,300-fold to homogeneity with an 8% yield from extracts of a recombinant strain of Pseudomonas denitrificans. Precorrin-6y was identified by fast atom bombardment-mass spectrometry. It was converted in high yield (90%) to hydrogenobyrinic acid by cell-free protein preparations from P. denitrificans. For the purification and characterization of precorrin-6x reductase, a coupled-enzyme radioenzymatic assay was developed in which precorrin-6y was methylated in situ by the cobL gene product (F. Blanche, A. Famechon, D. Thibaut, L. Debussche, B. Cameron, J. Crouzet, J. Bacteriol. 174:1050-1052, 1992) in the presence of [methyl-3H]S-adenosyl-L-methionine. Molecular weights of precorrin-6x reductase obtained by gel filtration (Mr congruent to 27,000) and by analytical sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Mr congruent to 31,000) were consistent with the enzyme being a monomer. Km values of 3.6 +/- 0.2 microM for precorrin-6x and 23.5 +/- 3.5 microM for NADPH and a Vmax value of 17,000 U mg-1 were obtained at pH 7.7. The N-terminal sequence (six amino acids) and three internal sequences obtained after tryptic digestion of the enzyme were determined by microsequencing and established that precorrin-6x reductase is encoded by the cobK gene, located on a previously described 8.7-kb EcoRI fragment (J. Crouzet, B. Cameron, L. Cauchois, S. Rigault, M.-C. Rouyez, F. Blanche, D. Thibaut, and L. Debussche, J. Bacteriol. 172:5980-5990, 1990). However, the coding sequence was shown to be on the strand complementary to the one previously proposed as the coding strand. Images PMID:1732193

  1. Reductive dechlorination of 2,4-dichlorobenzoate to 4-chlorobenzoate and hydrolytic dehalogenation of 4-chloro-, 4-bromo-, and 4-iodobenzoate by Alcaligenes denitrificans NTB-1.

    PubMed Central

    van den Tweel, W J; Kok, J B; de Bont, J A

    1987-01-01

    Alcaligenes denitrificans NTB-1, previously isolated on 4-chlorobenzoate, also utilized 4-bromo-, 4-iodo-, and 2,4-dichlorobenzoate but not 4-fluorobenzoate as a sole carbon and energy source. During growth, stoichiometric amounts of halide were released. Experiments with whole cells and cell extracts revealed that 4-bromo- and 4-iodobenzoate were metabolized like 4-chlorobenzoate, involving an initial hydrolytic dehalogenation yielding 4-hydroxybenzoate, which in turn was hydroxylated to 3,4-dihydroxybenzoate. The initial step in the metabolism of 2,4-dichlorobenzoate was catalyzed by a novel type of reaction for aerobic organisms, involving inducible reductive dechlorination to 4-chlorobenzoate. Under conditions of low and controlled oxygen concentrations, A. denitrificans NTB-1 converted all 4-halobenzoates and 2,4-dichlorobenzoate almost quantitatively to 4-hydroxybenzoate. PMID:3579283

  2. Treatment of naphthalene-2-sulfonic acid from tannery wastewater by a granular activated carbon fixed bed inoculated with bacterial isolates Arthrobacter globiformis and Comamonas testosteroni.

    PubMed

    Song, Zhi; Edwards, Suzanne R; Burns, Richard G

    2006-02-01

    The kinetics of naphthalene-2-sulfonic acid (2-NSA) adsorption by granular activated carbon (GAC) were measured and the relationships between adsorption, desorption, bioavailability and biodegradation assessed. The conventional Langmuir model fitted the experimental sorption isotherm data and introduced 2-NSA degrading bacteria, established on the surface of the GAC, did not interfere with adsorption. The potential value of GAC as a microbial support in the aerobic degradation of 2-NSA by Arthrobacter globiformis and Comamonas testosteroni was investigated. Using both virgin and microbially colonised GAC, adsorption removed 2-NSA from the liquid phase up to its saturation capacity of 140 mg/g GAC within 48 h. However, between 83.2% and 93.3% of the adsorbed 2-NSA was bioavailable to both bacterial species as a source of carbon for growth. In comparison to the non-inoculated GAC, the combination of rapid adsorption and biodegradation increased the amount (by 70-93%) of 2-NSA removal from the influent phase as well as the bed-life of the GAC (from 40 to >120 d). A microbially conditioned GAC fixed-bed reactor containing 15 g GAC removed 100% 2-NSA (100 mg/l) from tannery wastewater at an empty bed contact time of 22 min for a minimum of 120 d without the need for GAC reconditioning or replacement. This suggests that small volume GAC bioreactors could be used for tannery wastewater recycling.

  3. Ligand specificity of MobR, a transcriptional regulator for the 3-hydroxybenzoate hydroxylase gene of Comamonas testosteroni KH122-3s

    SciTech Connect

    Yoshida, Mariko; Hiromoto, Takeshi; Hosokawa, Keiichi; Yamaguchi, Hiroshi; Fujiwara, Shinsuke

    2007-10-19

    MobR from Comamonas testosteroni KH122-3s is a member of the MarR family of transcriptional regulators and functions as a repressor for the mobA gene that encodes a 3-hydroxybenzoate 4-hydroxylase. 3-Hydroxybenzoate binds to MobR as a ligand, resulting in an efficient induction of mobA. Various 3-hydroxybenzoate analogues were examined for their inducibilities using the mobA::lacZ transcriptional fusion system. {beta}-Galactosidase was induced by the addition of 2,3-dihydroxybenzoate or 3,5-dihydroxybenzoate besides 3-hydroxybenzoate, suggesting that the hydroxyl group at position 3 is critical in addition to the carboxyl group on the aromatic ring. A gel mobility-shift assay also showed that MobR was released from the target DNA in the presence of these compounds. Circular dichroism studies demonstrated that MobR adopted two conformational states corresponding to the 3-hydroxybenzoate-bound and unbound forms. Other ligands also induced the structural change as well; however, the tertiary structures of converted forms were different from those by 3-hydroxybenzoate.

  4. The Fe-type nitrile hydratase from Comamonas testosteroni Ni1 does not require an activator accessory protein for expression in Escherichia coli

    SciTech Connect

    Kuhn, Misty L.; Martinez, Salette; Gumataotao, Natalie; Bornscheuer, Uwe; Liu, Dali; Holz, Richard C.

    2012-10-10

    We report herein the functional expression of an Fe-type nitrile hydratase (NHase) without the co-expression of an activator protein or the Escherichia coli chaperone proteins GroES/EL. Soluble protein was obtained when the {alpha}- and {beta}-subunit genes of the Fe-type NHase Comamonas testosteroni Ni1 (CtNHase) were synthesized with optimized E. coli codon usage and co-expressed. As a control, the Fe-type NHase from Rhodococcus equi TG328-2 (ReNHase) was expressed with (ReNHase{sup +Act}) and without (ReNHase{sup -Act}) its activator protein, establishing that expression of a fully functional, metallated ReNHase enzyme requires the co-expression of its activator protein, similar to all other Fe-type NHase enzymes reported to date, whereas the CtNHase does not. The X-ray crystal structure of CtNHase was determined to 2.4 {angstrom} resolution revealing an {alpha}{beta} heterodimer, similar to other Fe-type NHase enzymes, except for two important differences. First, two His residues reside in the CtNHase active site that are not observed in other Fe-type NHase enzymes and second, the active site Fe(III) ion resides at the bottom of a wide solvent exposed channel. The solvent exposed active site, along with the two active site histidine residues, are hypothesized to play a role in iron incorporation in the absence of an activator protein.

  5. Fe(III)-enhanced anaerobic transformation of 2,4-dichlorophenoxyacetic acid by an iron-reducing bacterium Comamonas koreensis CY01.

    PubMed

    Wu, Chun-Yuan; Zhuang, Li; Zhou, Shun-Gui; Li, Fang-Bai; Li, Xiao-Min

    2010-01-01

    This work studied the ability of Comamonas koreensis CY01 to reduce Fe(III) (hydr)oxides by coupling the oxidation of electron donors and the enhanced biodegradation of 2,4-dichlorophenoxyacetic acid (2,4-D) by the presence of Fe(III) (hydr)oxides. The experimental results suggested that strain CY01 can utilize ferrihydrite, goethite, lepidocrocite or hematite as the terminal electron acceptor and citrate, glycerol, glucose or sucrose as the electron donor. Strain CY01 could transform 2,4-D to 4-chlorophenol through reductive side-chain removal and dechlorination. Under the anaerobic conditions, Fe(III) reduction and 2,4-D biodegradation by strain CY01 occurred simultaneously. The presence of Fe(III) (hydr)oxides would significantly enhance 2,4-D biodegradation, probably due to the fact that the reactive mineral-bound Fe(II) species generated from Fe(III) reduction can abiotically reduce 2,4-D. This is the first report of a strain of C. koreensis capable of reducing Fe(III) (hydr)oxides and 2,4-D, which extends the diversity of iron-reducing bacteria associated with dechlorination.

  6. Structure of a catalytic dimer of the α- and β-subunits of the F-ATPase from Paracoccus denitrificans at 2.3 Å resolution

    SciTech Connect

    Morales-Ríos, Edgar; Montgomery, Martin G.; Leslie, Andrew G. W.; García-Trejo, José J.; Walker, John E.

    2015-09-23

    The structure of the αβ heterodimer of the F-ATPase from the α-proteobacterium P. denitrificans has been determined at 2.3 Å resolution. It corresponds to the ‘open’ or ‘empty’ catalytic interface found in other F-ATPases. The structures of F-ATPases have predominantly been determined from mitochondrial enzymes, and those of the enzymes in eubacteria have been less studied. Paracoccus denitrificans is a member of the α-proteobacteria and is related to the extinct protomitochondrion that became engulfed by the ancestor of eukaryotic cells. The P. denitrificans F-ATPase is an example of a eubacterial F-ATPase that can carry out ATP synthesis only, whereas many others can catalyse both the synthesis and the hydrolysis of ATP. Inhibition of the ATP hydrolytic activity of the P. denitrificans F-ATPase involves the ζ inhibitor protein, an α-helical protein that binds to the catalytic F{sub 1} domain of the enzyme. This domain is a complex of three α-subunits and three β-subunits, and one copy of each of the γ-, δ- and ∊-subunits. Attempts to crystallize the F{sub 1}–ζ inhibitor complex yielded crystals of a subcomplex of the catalytic domain containing the α- and β-subunits only. Its structure was determined to 2.3 Å resolution and consists of a heterodimer of one α-subunit and one β-subunit. It has no bound nucleotides, and it corresponds to the ‘open’ or ‘empty’ catalytic interface found in other F-ATPases. The main significance of this structure is that it aids in the determination of the structure of the intact membrane-bound F-ATPase, which has been crystallized.

  7. Structure of the two-domain hexameric APS kinase from Thiobacillus denitrificans: structural basis for the absence of ATP sulfurylase activity

    SciTech Connect

    Gay, Sean C.; Segel, Irwin H.; Fisher, Andrew J.

    2009-10-01

    APS kinase from Thiobacillus denitrificans contains an inactive N-terminal ATP sulfurylase domain. The structure presented unveils the first hexameric assembly for an APS kinase, and reveals that structural changes in the N-terminal domain disrupt the ATP sulfurylase active site thus prohibiting activity. The Tbd-0210 gene of the chemolithotrophic bacterium Thiobacillus denitrificans is annotated to encode a 60.5 kDa bifunctional enzyme with ATP sulfurylase and APS kinase activity. This putative bifunctional enzyme was cloned, expressed and structurally characterized. The 2.95 Å resolution X-ray crystal structure reported here revealed a hexameric assembly with D{sub 3} symmetry. Each subunit contains a large N-terminal sulfurylase-like domain and a C-terminal APS kinase domain reminiscent of the two-domain fungal ATP sulfurylases of Penicillium chrysogenum and Saccharomyces cerevisiae, which also exhibit a hexameric assembly. However, the T. denitrificans enzyme exhibits numerous structural and sequence differences in the N-terminal domain that render it inactive with respect to ATP sulfurylase activity. Surprisingly, the C-terminal domain does indeed display APS kinase activity, indicating that this gene product is a true APS kinase. Therefore, these results provide the first structural insights into a unique hexameric APS kinase that contains a nonfunctional ATP sulfurylase-like domain of unknown function.

  8. Structure of a catalytic dimer of the α- and β-subunits of the F-ATPase from Paracoccus denitrificans at 2.3 Å resolution.

    PubMed

    Morales-Ríos, Edgar; Montgomery, Martin G; Leslie, Andrew G W; García-Trejo, José J; Walker, John E

    2015-10-01

    The structures of F-ATPases have predominantly been determined from mitochondrial enzymes, and those of the enzymes in eubacteria have been less studied. Paracoccus denitrificans is a member of the α-proteobacteria and is related to the extinct protomitochondrion that became engulfed by the ancestor of eukaryotic cells. The P. denitrificans F-ATPase is an example of a eubacterial F-ATPase that can carry out ATP synthesis only, whereas many others can catalyse both the synthesis and the hydrolysis of ATP. Inhibition of the ATP hydrolytic activity of the P. denitrificans F-ATPase involves the ζ inhibitor protein, an α-helical protein that binds to the catalytic F1 domain of the enzyme. This domain is a complex of three α-subunits and three β-subunits, and one copy of each of the γ-, δ- and ℇ-subunits. Attempts to crystallize the F1-ζ inhibitor complex yielded crystals of a subcomplex of the catalytic domain containing the α- and β-subunits only. Its structure was determined to 2.3 Å resolution and consists of a heterodimer of one α-subunit and one β-subunit. It has no bound nucleotides, and it corresponds to the `open' or `empty' catalytic interface found in other F-ATPases. The main significance of this structure is that it aids in the determination of the structure of the intact membrane-bound F-ATPase, which has been crystallized.

  9. Isolation and characterization of ubiquinol oxidase complexes from Paracoccus denitrificans cells cultured under various limiting growth conditions in the chemostat.

    PubMed

    Bosma, G; Braster, M; Stouthamer, A H; van Verseveld, H W

    1987-06-15

    To obtain more information about the composition of the respiratory chain under different growth conditions and about the regulation of electron-transfer to several oxidases and reductases, ubiquinol oxidase complexes were partially purified from membranes of Paracoccus denitrificans cells grown in carbon-source-limited aerobic, nitrate-limited anaerobic and oxygen-limited chemostat cultures. The isolated enzymes consisted of cytochromes bc1, c552 and aa3. In comparison with the aerobic ubiquinol oxidase complex, the oxygen- and nitrate-limited ones contained, respectively, less and far less of the cytochrome aa3 subunits and the anaerobic complex also contained lower amounts of cytochrome c552. In addition, extra haem-containing polypeptides were present with apparent Mr of 14,000, 30,000 and 45,000, the former one only in the anaerobic and the latter two in both the anaerobic and oxygen-limited preparations. This is the first report describing four different membrane-bound c-type cytochromes. The potentiometric and spectral characteristics of the redox components in membrane particles and isolated ubiquinol oxidase fractions were determined by combined potentiometric analysis and spectrum deconvolution. Membranes of nitrate- and oxygen-limited cells contained extra high-potential cytochrome b in comparison with the membranes of aerobically grown cells. No difference was detected between the three isolated ubiquinol oxidase complexes. Aberrances with already published values of redox potentials are discussed. PMID:3036512

  10. Atomic resolution structure of pseudoazurin from the methylotrophic denitrifying bacterium Hyphomicrobium denitrificans: structural insights into its spectroscopic properties.

    PubMed

    Hira, Daisuke; Nojiri, Masaki; Suzuki, Shinnichiro

    2009-01-01

    The crystal structure of native pseudoazurin (HdPAz) from the methylotrophic denitrifying bacterium Hyphomicrobium denitrificans has been determined at a resolution of 1.18 A. After refinement with SHELX employing anisotropic displacement parameters and riding H atoms, R(work) and R(free) were 0.135 and 0.169, respectively. Visualization of the anisotropic displacement parameters as thermal ellipsoids provided insight into the atomic motion within the perturbed type 1 Cu site. The asymmetric unit includes three HdPAz molecules which are tightly packed by head-to-head cupredoxin dimer formation. The shape of the Cu-atom ellipsoid implies significant vibrational motion diagonal to the equatorial xy plane defined by the three ligands (two His and one Cys). The geometric parameters of the type 1 Cu site in the HdPAz structure differ unambiguously from those of other pseudoazurins. It is demonstrated that their structural aspects are consistent with the unique visible absorption spectrum. PMID:19153470

  11. AztD, a Periplasmic Zinc Metallochaperone to an ATP-binding Cassette (ABC) Transporter System in Paracoccus denitrificans.

    PubMed

    Handali, Melody; Roychowdhury, Hridindu; Neupane, Durga P; Yukl, Erik T

    2015-12-11

    Bacterial ATP-binding cassette (ABC) transporters of transition metals are essential for acquisition of necessary elements from the environment. A large number of Gram-negative bacteria, including human pathogens, have a fourth conserved gene of unknown function adjacent to the canonical permease, ATPase, and solute-binding protein (SBP) genes of the AztABC zinc transporter system. To assess the function of this putative accessory factor (AztD) from Paracoccus denitrificans, we have analyzed its transcriptional regulation, metal binding properties, and interaction with the SBP (AztC). Transcription of the aztD gene is significantly up-regulated under conditions of zinc starvation. Recombinantly expressed AztD purifies with slightly substoichiometric zinc from the periplasm of Escherichia coli and is capable of binding up to three zinc ions with high affinity. Size exclusion chromatography and a simple intrinsic fluorescence assay were used to determine that AztD as isolated is able to transfer bound zinc nearly quantitatively to apo-AztC. Transfer occurs through a direct, associative mechanism that prevents loss of metal to the solvent. These results indicate that AztD is a zinc chaperone to AztC and likely functions to maintain zinc homeostasis through interaction with the AztABC system. This work extends our understanding of periplasmic zinc trafficking and the function of chaperones in this process.

  12. Achromobacter denitrificans Strain YD35 Pyruvate Dehydrogenase Controls NADH Production To Allow Tolerance to Extremely High Nitrite Levels

    PubMed Central

    Doi, Yuki; Shimizu, Motoyuki; Fujita, Tomoya; Nakamura, Akira; Takizawa, Noboru

    2014-01-01

    We identified the extremely nitrite-tolerant bacterium Achromobacter denitrificans YD35 that can grow in complex medium containing 100 mM nitrite (NO2−) under aerobic conditions. Nitrite induced global proteomic changes and upregulated tricarboxylate (TCA) cycle enzymes as well as antioxidant proteins in YD35. Transposon mutagenesis generated NO2−-hypersensitive mutants of YD35 that had mutations at genes for aconitate hydratase and α-ketoglutarate dehydrogenase in the TCA cycle and a pyruvate dehydrogenase (Pdh) E1 component, indicating the importance of TCA cycle metabolism to NO2− tolerance. A mutant in which the pdh gene cluster was disrupted (Δpdh mutant) could not grow in the presence of 100 mM NO2−. Nitrite decreased the cellular NADH/NAD+ ratio and the cellular ATP level. These defects were more severe in the Δpdh mutant, indicating that Pdh contributes to upregulating cellular NADH and ATP and NO2−-tolerant growth. Exogenous acetate, which generates acetyl coenzyme A and then is metabolized by the TCA cycle, compensated for these defects caused by disruption of the pdh gene cluster and those caused by NO2−. These findings demonstrate a link between NO2− tolerance and pyruvate/acetate metabolism through the TCA cycle. The TCA cycle mechanism in YD35 enhances NADH production, and we consider that this contributes to a novel NO2−-tolerating mechanism in this strain. PMID:24413603

  13. Growth-Associated Production of Poly(3-Hydroxyvalerate) from n-Pentanol by a Methylotrophic Bacterium, Paracoccus denitrificans

    PubMed Central

    Yamane, T.; Chen, X.; Ueda, S.

    1996-01-01

    Paracoccus denitrificans accumulated a polyester in its cells during growth on n-pentanol. The composition of the polyester varied during the cultivation: the level of the 3-hydroxyvalerate unit in the polyester increased, and eventually a homopolymeric poly(3-hydroxyvalerate) [P(3HV)] accumulated to an amount 22 to 24% of the cell dry weight. Growth-associated polyester synthesis was considerably affected by n-pentanol when its concentration was controlled at several levels. Maximum accumulation of the polyester was obtained at 0.02% (vol/vol). Physical and mechanical characteristics of the P(3HV) were determined and compared with those of other homo- and copolyesters. The P(3HV) was dextrorotatory and had number-averaged and weight-averaged molecular masses of 128,000 and 888,000 Da, respectively, with a rate of polydispersity of 6.93. The level of tensile strength of the P(3HV) was lower, and its extension to break was higher than that of the poly(3-hydroxybutyrate) homopolyester. PMID:16535226

  14. Transcriptional Regulation, Metal Binding Properties and Structure of Pden1597, an Unusual Zinc Transport Protein from Paracoccus denitrificans.

    PubMed

    Handali, Melody; Neupane, Durga P; Roychowdhury, Hridindu; Yukl, Erik T

    2015-05-01

    ATP-binding cassette (ABC) transporters of the cluster 9 family are ubiquitous among bacteria and essential for acquiring Zn(2+) and Mn(2+) from the environment or, in the case of pathogens, from the host. These rely on a substrate-binding protein (SBP) to coordinate the relevant metal with high affinity and specificity and subsequently release it to a membrane permease for translocation into the cytoplasm. Although a number of cluster 9 SBP structures have been determined, the structural attributes conferring Zn(2+) or Mn(2+) specificity remain ambiguous. Here we describe the gene expression profile, in vitro metal binding properties, and crystal structure of a new cluster 9 SBP from Paracoccus denitrificans we have called AztC. Although all of our results strongly indicate Zn(2+) over Mn(2+) specificity, the Zn(2+) ion is coordinated by a conserved Asp residue only observed to date as a metal ligand in Mn(2+)-specific SBPs. The unusual sequence properties of this protein are shared among close homologues, including members from the human pathogens Klebsiella pneumonia and Enterobacter aerogenes, and would seem to suggest a subclass of Zn(2+)-specific transporters among the cluster 9 family. In any case, the unusual coordination environment of AztC expands the already considerable range of those available to Zn(2+)-specific SBPs and highlights the presence of a His-rich loop as the most reliable indicator of Zn(2+) specificity.

  15. Transcriptional Regulation, Metal Binding Properties and Structure of Pden1597, an Unusual Zinc Transport Protein from Paracoccus denitrificans.

    PubMed

    Handali, Melody; Neupane, Durga P; Roychowdhury, Hridindu; Yukl, Erik T

    2015-05-01

    ATP-binding cassette (ABC) transporters of the cluster 9 family are ubiquitous among bacteria and essential for acquiring Zn(2+) and Mn(2+) from the environment or, in the case of pathogens, from the host. These rely on a substrate-binding protein (SBP) to coordinate the relevant metal with high affinity and specificity and subsequently release it to a membrane permease for translocation into the cytoplasm. Although a number of cluster 9 SBP structures have been determined, the structural attributes conferring Zn(2+) or Mn(2+) specificity remain ambiguous. Here we describe the gene expression profile, in vitro metal binding properties, and crystal structure of a new cluster 9 SBP from Paracoccus denitrificans we have called AztC. Although all of our results strongly indicate Zn(2+) over Mn(2+) specificity, the Zn(2+) ion is coordinated by a conserved Asp residue only observed to date as a metal ligand in Mn(2+)-specific SBPs. The unusual sequence properties of this protein are shared among close homologues, including members from the human pathogens Klebsiella pneumonia and Enterobacter aerogenes, and would seem to suggest a subclass of Zn(2+)-specific transporters among the cluster 9 family. In any case, the unusual coordination environment of AztC expands the already considerable range of those available to Zn(2+)-specific SBPs and highlights the presence of a His-rich loop as the most reliable indicator of Zn(2+) specificity. PMID:25787075

  16. Achromobacter denitrificans SP1 produces pharmaceutically active 25C prodigiosin upon utilizing hazardous di(2-ethylhexyl)phthalate.

    PubMed

    Pradeep, S; Sarath Josh, M K; Balachandran, S; Sudha Devi, R; Sadasivam, R; Thirugnanam, P E; Doble, Mukesh; Anderson, Robin C; Benjamin, Sailas

    2014-11-01

    This first report describes the purification and identification of an orange-red pigment produced by Achromobacter denitrificans strain SP1 (isolated from sewage sludge heavily contaminated with plastics) during its growth in a simple basal salt medium supplemented with the hazardous di(2-ethylhexyl)phthalate (DEHP) blended in PVC blood bag (in situ) or free DEHP (ex situ) as carbon source. The cell-bound pigment was elucidated, characterized at molecular level, and described as an unusual 25C prodigiosin analog for the first time. At laboratory conditions (in flasks), the dry cell mass was 75.2mg/g blood bag, which upon extraction yielded 7.1mg prodigiosin; at this stage the pH of the medium was dropped from 7.2 to 3.5. Considering its pharmaceutical importance, taking 10 known prodigiosins as controls, this 25C prodigiosin was subjected to molecular docking studies, showed comparable and promising binding efficiencies with the crucial molecular human targets like cycloxygenase-2, ZAP-70 kinase and Jak-3 kinase.

  17. Structure of a catalytic dimer of the α- and β-subunits of the F-ATPase from Paracoccus denitrificans at 2.3 Å resolution

    PubMed Central

    Morales-Ríos, Edgar; Montgomery, Martin G.; Leslie, Andrew G. W.; García-Trejo, José J.; Walker, John E.

    2015-01-01

    The structures of F-ATPases have predominantly been determined from mitochondrial enzymes, and those of the enzymes in eubacteria have been less studied. Paracoccus denitrificans is a member of the α-proteobacteria and is related to the extinct protomitochondrion that became engulfed by the ancestor of eukaryotic cells. The P. denitrificans F-ATPase is an example of a eubacterial F-ATPase that can carry out ATP synthesis only, whereas many others can catalyse both the synthesis and the hydrolysis of ATP. Inhibition of the ATP hydrolytic activity of the P. denitrificans F-ATPase involves the ζ inhibitor protein, an α-helical protein that binds to the catalytic F1 domain of the enzyme. This domain is a complex of three α-subunits and three β-subunits, and one copy of each of the γ-, δ- and ∊-subunits. Attempts to crystallize the F1–ζ inhibitor complex yielded crystals of a subcomplex of the catalytic domain containing the α- and β-subunits only. Its structure was determined to 2.3 Å resolution and consists of a heterodimer of one α-subunit and one β-subunit. It has no bound nucleotides, and it corresponds to the ‘open’ or ‘empty’ catalytic interface found in other F-ATPases. The main significance of this structure is that it aids in the determination of the structure of the intact membrane-bound F-ATPase, which has been crystallized. PMID:26457523

  18. Identification and characterization of a novel translational repressor of the steroid-inducible 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase gene in Comamonas testosteroni.

    PubMed

    Xiong, Guangming; Martin, Hans-Jörg; Maser, Edmund

    2003-11-28

    Comamonas testosteroni 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase (3 alpha-HSD/CR) is a key enzyme in the degradation of steroid compounds in soil and may therefore play a significant role in the bioremediation of hormonally active compounds in the environment. The enzyme is also involved in the degradation of the steroid antibiotic fusidic acid. In addition, 3 alpha-HSD/CR mediates the carbonyl reduction of non-steroidal aldehydes and ketones. Because the gene of 3 alpha-HSD/CR (hsdA) is inducible by steroids, we were interested in the mode of its molecular regulation. Recently, we could identify the first molecular determinant in procaryotic steroid signaling, i.e. a repressor protein (RepA), which acts as a negative regulator by binding to upstream operator sequences of hsdA, thereby blocking hsdA transcription. In this work, we identified and cloned a second novel regulator gene that we named repB. The gene locates 932 bp downstream from hsdA on the C. testosteroni chromosome with an orientation opposite to that of hsdA. The open reading frame of repB consists of 237 bp and translates into a protein of 78 amino acids that was found to act as a repressor that regulates hsdA expression on the translational level. Northern blot analysis, UV-cross linking, gel-shift assays, and competition experiments proved that RepB binds to a 16-nucleotide sequence downstream of AUG at the 5' end of the 3 alpha-HSD/CR mRNA, thereby blocking hsdA translation. Testosterone, on the other hand, was shown to specifically bind to RepB, thereby yielding the release of RepB from the 3 alpha-HSD/CR mRNA such that hsdA translation could proceed. Data bank searches with the RepB primary structure yielded a 46.2% identity to the regulator of nucleoside diphosphate kinase, a formerly unknown protein from Escherichia coli that can restore a growth defect in alginate production in Pseudomonas aeruginosa. In conclusion, the induction of hsdA by steroids in fact is a derepression

  19. Whole-Genome Transcriptional Analysis of Chemolithoautotrophic Thiosulfate Oxidation by Thiobacillus denitrificans Under Aerobic vs. Denitrifying Conditions

    SciTech Connect

    Beller, H R; Letain, T E; Chakicherla, A; Kane, S R; Legler, T C; Coleman, M A

    2006-04-22

    Thiobacillus denitrificans is one of the few known obligate chemolithoautotrophic bacteria capable of energetically coupling thiosulfate oxidation to denitrification as well as aerobic respiration. As very little is known about the differential expression of genes associated with ke chemolithoautotrophic functions (such as sulfur-compound oxidation and CO2 fixation) under aerobic versus denitrifying conditions, we conducted whole-genome, cDNA microarray studies to explore this topic systematically. The microarrays identified 277 genes (approximately ten percent of the genome) as differentially expressed using Robust Multi-array Average statistical analysis and a 2-fold cutoff. Genes upregulated (ca. 6- to 150-fold) under aerobic conditions included a cluster of genes associated with iron acquisition (e.g., siderophore-related genes), a cluster of cytochrome cbb3 oxidase genes, cbbL and cbbS (encoding the large and small subunits of form I ribulose 1,5-bisphosphate carboxylase/oxygenase, or RubisCO), and multiple molecular chaperone genes. Genes upregulated (ca. 4- to 95-fold) under denitrifying conditions included nar, nir, and nor genes (associated respectively with nitrate reductase, nitrite reductase, and nitric oxide reductase, which catalyze successive steps of denitrification), cbbM (encoding form II RubisCO), and genes involved with sulfur-compound oxidation (including two physically separated but highly similar copies of sulfide:quinone oxidoreductase and of dsrC, associated with dissimilatory sulfite reductase). Among genes associated with denitrification, relative expression levels (i.e., degree of upregulation with nitrate) tended to decrease in the order nar > nir > nor > nos. Reverse transcription, quantitative PCR analysis was used to validate these trends.

  20. Mechanism of nitrogenase switch-off by oxygen. [Klebsiella pneumoniae; Rhodopseudomonas sphaeroides f. sp. denitrificans; Rhodopseudomonas capsulate

    SciTech Connect

    Goldberg, I.; Nadler, V.; Hochman, A.

    1987-02-01

    Oxygen caused a reversible inhibition (switch-off) of nitrogenase activity in whole cells of four strains of diazotrophs, the facultative anaerobe Klebsiella pneumoniae and three strains of photosynthetic bacteria (Rhodopseudomonas sphaeroides f. sp. denitrificans and Rhodopseudomonas capsulata strians AD2 and BK5). In K. pneumoniae 50% inhibition of acetylene reduction was attained at an O/sub 2/ concentration of 0.37 ..mu..M. Cyanide (90 ..mu..M), which did not affect acetylene reduction but inhibited whole-cell respiration by 60 to 70%, shifted the O/sub 2/ concentration that caused 50% inhibition of nitrogenase activity to 2.9 ..mu..M. A mutant strain of K. pneumoniae, strain AH11, has a respiration rate that is 65 to 75% higher than that of the wild type, but is nitrogenase activity is similar to wild-type activity. Acetylene reduction by whole cells of this mutant was inhibited 50% by 0.20 ..mu..M O/sub 2/. Inhibition by CN/sup -/ of 40 to 50% of the O/sub 2/ uptake in the mutant shifted the O/sub 2/ concentration that caused 50% inhibition of nitrogenase to 1.58 ..mu..M. Thus, when the respiration rates were lower, higher oxygen concentrations were required to inhibit nitrogenase. Reversible inhibition of nitrogenase activity in vivo was caused under anaerobic conditions by other electron acceptors. Addition of 2 mM sulfite to cell suspensions of R. capsulata B10 and R. sphaeroides inhibited nitrogenase activity. Nitrite also inhibited acetylene reduction in whole cells of the photodenitrifier R. sphaeroides but not in R. capsulata B10, which is not capable of enzymatic reduction of NO/sub 2//sup -/. Lower concentrations of NO/sub 2//sup -/ were required to inhibit the activity in NO/sub 3//sup -/-grown cells, which have higher activities of nitrite reductase.

  1. Structure of ATP synthase from Paracoccus denitrificans determined by X-ray crystallography at 4.0 Å resolution

    PubMed Central

    Morales-Rios, Edgar; Montgomery, Martin G.; Leslie, Andrew G. W.; Walker, John E.

    2015-01-01

    The structure of the intact ATP synthase from the α-proteobacterium Paracoccus denitrificans, inhibited by its natural regulatory ζ-protein, has been solved by X-ray crystallography at 4.0 Å resolution. The ζ-protein is bound via its N-terminal α-helix in a catalytic interface in the F1 domain. The bacterial F1 domain is attached to the membrane domain by peripheral and central stalks. The δ-subunit component of the peripheral stalk binds to the N-terminal regions of two α-subunits. The stalk extends via two parallel long α-helices, one in each of the related b and b′ subunits, down a noncatalytic interface of the F1 domain and interacts in an unspecified way with the a-subunit in the membrane domain. The a-subunit lies close to a ring of 12 c-subunits attached to the central stalk in the F1 domain, and, together, the central stalk and c-ring form the enzyme’s rotor. Rotation is driven by the transmembrane proton-motive force, by a mechanism where protons pass through the interface between the a-subunit and c-ring via two half-channels in the a-subunit. These half-channels are probably located in a bundle of four α-helices in the a-subunit that are tilted at ∼30° to the plane of the membrane. Conserved polar residues in the two α-helices closest to the c-ring probably line the proton inlet path to an essential carboxyl group in the c-subunit in the proton uptake site and a proton exit path from the proton release site. The structure has provided deep insights into the workings of this extraordinary molecular machine. PMID:26460036

  2. Electronic and vibrational spectroscopy of the cytochrome c:cytochrome c oxidase complexes from bovine and Paracoccus denitrificans.

    PubMed Central

    Lynch, S. R.; Copeland, R. A.

    1992-01-01

    The 1:1 complex between horse heart cytochrome c and bovine cytochrome c oxidase, and between yeast cytochrome c and Paracoccus denitrificans cytochrome c oxidase have been studied by a combination of second derivative absorption, circular dichroism (CD), and resonance Raman spectroscopy. The second derivative absorption and CD spectra reveal changes in the electronic transitions of cytochrome a upon complex formation. These results could reflect changes in ground state heme structure or changes in the protein environment surrounding the chromophore that affect either the ground or excited electronic states. The resonance Raman spectrum, on the other hand, reflects the heme structure in the ground electronic state only and shows no significant difference between cytochrome a vibrations in the complex or free enzyme. The only major difference between the Raman spectra of the free enzyme and complex is a broadening of the cytochrome a3 formyl band of the complex that is relieved upon complex dissociation at high ionic strength. These data suggest that the differences observed in the second derivative and CD spectra are the result of changes in the protein environment around cytochrome a that affect the electronic excited state. By analogy to other protein-chromophore systems, we suggest that the energy of the Soret pi* state of cytochrome a may be affected by (1) changes in the local dielectric, possibly brought about by movement of a charged amino acid side chain in proximity to the heme group, or (2) pi-pi interactions between the heme and aromatic amino acid residues. PMID:1338946

  3. Nisaea denitrificans gen. nov., sp. nov. and Nisaea nitritireducens sp. nov., two novel members of the class Alphaproteobacteria from the Mediterranean Sea.

    PubMed

    Urios, Laurent; Michotey, Valérie; Intertaglia, Laurent; Lesongeur, Françoise; Lebaron, Philippe

    2008-10-01

    Two novel Gram-negative bacteria, designated strains DR41_21(T) and DR41_18(T), were isolated from coastal, surface waters of the north-western Mediterranean Sea. The cells were motile, pleomorphic rods, 2.9 microm long and 0.9 microm wide and formed cream colonies on marine agar medium. The G+C content of the genomic DNA was 60 mol%. Phylogenetic analysis of 16S rRNA gene sequences positioned the isolates in the class Alphaproteobacteria within the family Rhodospirillaceae. The 16S rRNA gene sequence similarity of the two strains was 98.8 % but DNA-DNA hybridization indicated only 55 % relatedness. Strain DR41_21(T) was able to denitrify and possessed nirK and nosZ genes, unlike strain DR41_18(T), which possessed only nirK. These isolates represent two novel species of a new genus, Nisaea gen. nov., for which the names Nisaea denitrificans sp. nov. and Nisaea nitritireducens sp. nov. are proposed. The type strain of Nisaea denitrificans is DR41_21(T) (=DSM 18348(T)=CIP 109265(T)=OOB 129(T)) and the type strain of Nisaea nitritireducens is DR41_18(T) (=DSM 19540(T)=CIP 109601(T)=OOB 128(T)).

  4. Structure of the two-domain hexameric APS kinase from Thiobacillus denitrificans: structural basis for the absence of ATP sulfurylase activity.

    PubMed

    Gay, Sean C; Segel, Irwin H; Fisher, Andrew J

    2009-10-01

    The Tbd_0210 gene of the chemolithotrophic bacterium Thiobacillus denitrificans is annotated to encode a 60.5 kDa bifunctional enzyme with ATP sulfurylase and APS kinase activity. This putative bifunctional enzyme was cloned, expressed and structurally characterized. The 2.95 A resolution X-ray crystal structure reported here revealed a hexameric assembly with D(3) symmetry. Each subunit contains a large N-terminal sulfurylase-like domain and a C-terminal APS kinase domain reminiscent of the two-domain fungal ATP sulfurylases of Penicillium chrysogenum and Saccharomyces cerevisiae, which also exhibit a hexameric assembly. However, the T. denitrificans enzyme exhibits numerous structural and sequence differences in the N-terminal domain that render it inactive with respect to ATP sulfurylase activity. Surprisingly, the C-terminal domain does indeed display APS kinase activity, indicating that this gene product is a true APS kinase. Therefore, these results provide the first structural insights into a unique hexameric APS kinase that contains a nonfunctional ATP sulfurylase-like domain of unknown function.

  5. Models for molybdenum coordination during the catalytic cycle of periplasmic nitrate reductase from Paracoccus denitrificans derived from EPR and EXAFS spectroscopy.

    PubMed

    Butler, C S; Charnock, J M; Bennett, B; Sears, H J; Reilly, A J; Ferguson, S J; Garner, C D; Lowe, D J; Thomson, A J; Berks, B C; Richardson, D J

    1999-07-13

    The periplasmic nitrate reductase from Paracoccus denitrificans is a soluble two-subunit enzyme which binds two hemes (c-type), a [4Fe-4S] center, and a bis molybdopterin guanine dinucleotide cofactor (bis-MGD). A catalytic cycle for this enzyme is presented based on a study of these redox centers using electron paramagnetic resonance (EPR) and extended X-ray absorption fine structure (EXAFS) spectroscopies. The Mo(V) EPR signal of resting NAP (High g [resting]) has g(av) = 1.9898 is rhombic, exhibits low anisotropy, and is split by two weakly interacting protons which are not solvent-exchangeable. Addition of exogenous ligands to this resting state (e.g., nitrate, nitrite, azide) did not change the form of the signal. A distinct form of the High g Mo(V) signal, which has slightly lower anisotropy and higher rhombicity, was trapped during turnover of nitrate and may represent a catalytically relevant Mo(V) intermediate (High g [nitrate]). Mo K-edge EXAFS analysis was undertaken on the ferricyanide oxidized enzyme, a reduced sample frozen within 10 min of dithionite addition, and a nitrate-reoxidized form of the enzyme. The oxidized enzyme was fitted best as a di-oxo Mo(VI) species with 5 sulfur ligands (4 at 2. 43 A and 1 at 2.82 A), and the reduced form was fitted best as a mono-oxo Mo(IV) species with 3 sulfur ligands at 2.35 A. The addition of nitrate to the reduced enzyme resulted in reoxidation to a di-oxo Mo(VI) species similar to the resting enzyme. Prolonged incubation of NAP with dithionite in the absence of nitrate (i.e., nonturnover conditions) resulted in the formation of a species with a Mo(V) EPR signal that is quite distinct from the High g family and which has a g(av) = 1.973 (Low g [unsplit]). This signal resembles those of the mono-MGD xanthine oxidase family and is proposed to arise from an inactive form of the nitrate reductase in which the Mo(V) form is only coordinated by the dithiolene of one MGD. In samples of NAP that had been reduced with

  6. Structure of the puf operon of the obligately aerobic, bacteriochlorophyll alpha-containing bacterium Roseobacter denitrificans OCh114 and its expression in a Rhodobacter capsulatus puf puc deletion mutant.

    PubMed Central

    Kortlüke, C; Breese, K; Gad'on, N; Labahn, A; Drews, G

    1997-01-01

    Roseobacter denitrificans (Erythrobacter species strain OCh114) synthesizes bacteriochlorophyll a (BChl) and the photosynthetic apparatus only in the presence of oxygen and is unable to carry out primary photosynthetic reactions and to grow photosynthetically under anoxic conditions. The puf operon of R. denitrificans has the same five genes in the same order as in many photosynthetic bacteria, i.e., pufBALMC. PufC, the tetraheme subunit of the reaction center (RC), consists of 352 amino acids (Mr, 39,043); 20 and 34% of the total amino acids are identical to those of PufC of Chloroflexus aurantiacus and Rubrivivax gelatinosus, respectively. The N-terminal hydrophobic domain is probably responsible for anchoring the subunit in the membrane. Four heme-binding domains are homologous to those of PufC in several purple bacteria. Sequences similar to pufQ and pufX of Rhodobacter capsulatus were not detected on the chromosome of R. denitrificans. The puf operon of R. denitrificans was expressed in trans in Escherichia coli, and all gene products were synthesized. The Roseobacter puf operon was also expressed in R. capsulatus CK11, a puf puc double-deletion mutant. For the first time, an RC/light-harvesting complex I core complex was heterologously synthesized. The strongest expression of the R. denitrificans puf operon was observed under the control of the R. capsulatus puf promoter, in the presence of pufQ and pufX and in the absence of pufC. Charge recombination between the primary donor P+ and the primary ubiquinone Q(A)- was observed in the transconjugant, showing that the M and L subunits of the RC were correctly assembled. The transconjugants did not grow photosynthetically under anoxic conditions. PMID:9286973

  7. Uptake of benzoic acid and chloro-substituted benzoic acids by alcaligenes denitrificans BRI 3010 and BRI 6011

    SciTech Connect

    Miguez, C.B.; Ingram, J.M.; MacLeod, R.A.

    1995-12-01

    The mechanism of uptake of benzoic and 2,4-dichlorobenzoic acid (2,4-DCBA) by Alcaligenes denitrificans BRI 3010 and BRI 6011 and Pseudomonas sp. strain B13, three organisms capable of degrading isomers of chlorinated benzoic acids, was investigated. In all three organisms, uptake of benzoic acid was inducible. For benzoic acid uptake into BRI 3010, monophasic saturation kinetics with apparent K{sub m} and V{sub max} values of 1.4 {mu}M and 3.2 nmol/min/mg of cell dry weight, respectively, were obtained. For BRI 6011, biphasic saturation kinetics were observed, suggesting presence of two uptake systems for benzoic acid with distinct K{sub m} (0.72 and 5.3 {mu}M) and V{sub max} (3.3 and 4.6 nmol/min/mg of cell dry weight) values. BRI 3010 and BRI 6011 accumulated benzoic acid against a concentration gradient by a factor of 8 and 10, respectively. A wide range of structural analogs, at 50-fold excess concentrations, inhibited benzoic acid uptake by BRI 3010 and BRI 6011, whereas with B13, only 3-chlorobenzoic acid was an effective inhibitor. For BRI 3010 and BRI 6011, the inhibition by the structural analogs was not of a competitive nature. Uptake of benzoic acid by BRI 3010 and BRI 6011 was inhibited by KCN, by the protonophore 3,5,3`, 4`-tetrachlorosalicylanilide (TCS), and, for BRI 6011, by anaerobiosis unless nitrate was present, thus indicating that energy was required for the uptake process. Uptake of 2,4-DCBA by BRI 6011 was constitutive and saturation uptake kinetics were not observed. Uptake of 2,4-DCBA by BRI 6011 was inhibited by KCN, TCS, and anaerobiosis even if nitrate was present, but the compound was not accumulated intracellularly against a concentration gradient. Uptake of 2,4-DCBA by BRI 6011 appears to occur by passive diffusion into the cell down its concentration gradient, which is maintained by the intracellular metabolism of the compound. This process could play an important role in the degradation of xenobiotic compounds by microorganisms.

  8. High crystallizability under air-exclusion conditions of the full-length LysR-type transcriptional regulator TsaR from Comamonas testosteroni T-2 and data-set analysis for a MIRAS structure-solution approach

    PubMed Central

    Monferrer, Dominique; Tralau, Tewes; Kertesz, Michael A.; Panjikar, Santosh; Usón, Isabel

    2008-01-01

    The full-length LysR-type transcriptional regulator TsaR from Comamonas testosteroni T-2 was heterologously overexpressed in Escherichia coli, purified and stabilized under conditions that favoured its rapid crystallization using the microbatch-under-oil technique. The purified protein was highly crystallizable and two different crystal forms were readily obtained. However, only monoclinic crystals gave diffraction beyond 2 Å and there was a slight variation in unit-cell parameters between crystals. The only other LysR-type regulator for which a full-length crystal form is available is CbnR, but no solution could be obtained when this was used as a model in molecular replacement. Mercury and xenon derivatives were therefore produced in order to phase the structure using a MIRAS approach. PMID:18678953

  9. Cloning and characterization of sulfite dehydrogenase, two c-type cytochromes, and a flavoprotein of Paracoccus denitrificans GB17: essential role of sulfite dehydrogenase in lithotrophic sulfur oxidation.

    PubMed

    Wodara, C; Bardischewsky, F; Friedrich, C G

    1997-08-01

    A 13-kb genomic region of Paracoccus dentrificans GB17 is involved in lithotrophic thiosulfate oxidation. Adjacent to the previously reported soxB gene (C. Wodara, S. Kostka, M. Egert, D. P. Kelly, and C. G. Friedrich, J. Bacteriol. 176:6188-6191, 1994), 3.7 kb were sequenced. Sequence analysis revealed four additional open reading frames, soxCDEF. soxC coded for a 430-amino-acid polypeptide with an Mr of 47,339 that included a putative signal peptide of 40 amino acids (Mr of 3,599) with a RR motif present in periplasmic proteins with complex redox centers. The mature soxC gene product exhibited high amino acid sequence similarity to the eukaryotic molybdoenzyme sulfite oxidase and to nitrate reductase. We constructed a mutant, GBsoxC delta, carrying an in-frame deletion in soxC which covered a region possibly coding for the molybdenum cofactor binding domain. GBsoxC delta was unable to grow lithoautotrophically with thiosulfate but grew well with nitrate as a nitrogen source or as an electron acceptor. Whole cells and cell extracts of mutant GBsoxC delta contained 10% of the thiosulfate-oxidizing activity of the wild type. Only a marginal rate of sulfite-dependent cytochrome c reduction was observed from cell extracts of mutant GBsoxC delta. These results demonstrated that sulfite dehydrogenase was essential for growth with thiosulfate of P. dentrificans GB17. soxD coded for a periplasmic diheme c-type cytochrome of 384 amino acids (Mr of 39,983) containing a putative signal peptide with an Mr of 2,363. soxE coded for a periplasmic monoheme c-type cytochrome of 236 amino acids (Mr of 25,926) containing a putative signal peptide with an Mr of 1,833. SoxD and SoxE were highly identical to c-type cytochromes of P. denitrificans and other organisms. soxF revealed an incomplete open reading frame coding for a peptide of 247 amino acids with a putative signal peptide (Mr of 2,629). The deduced amino acid sequence of soxF was 47% identical and 70% similar to the sequence

  10. A New Bacterial Steroid Degradation Gene Cluster in Comamonas testosteroni TA441 Which Consists of Aromatic-Compound Degradation Genes for Seco-Steroids and 3-Ketosteroid Dehydrogenase Genes

    PubMed Central

    Horinouchi, Masae; Hayashi, Toshiaki; Yamamoto, Takako; Kudo, Toshiaki

    2003-01-01

    In Comamonas testosteroni TA441, testosterone is degraded via aromatization of the A ring, which is cleaved by the meta-cleavage enzyme TesB, and further degraded by TesD, the hydrolase for the product of TesB. TesEFG, encoded downstream of TesD, are probably hydratase, aldolase, and dehydrogenase for degradation of 2-oxohex-4-enoicacid, one of the products of TesD. Here we present a new and unique steroid degradation gene cluster in TA441, which consists of ORF18, ORF17, tesI, tesH, ORF11, ORF12, and tesDEFG. TesH and TesI are 3-ketosteroid-Δ1-dehydrogenase and 3-ketosteroid-Δ4(5α)-dehydrogenase, respectively, which work in the early steps of steroid degradation. ORF17 probably encodes the reductase component of 9α-hydroxylase for 1,4-androstadiene-3,17-dione, which is the product of TesH in testosterone degradation. Gene disruption experiments showed that these genes are necessary for steroid degradation and do not have any isozymes in TA441. By Northern blot analysis, these genes were shown to be induced when TA441 was incubated with steroids (testosterone and cholic acid) but not with aromatic compounds [phenol, biphenyl, and 3-(3-hydroxyphenyl)propionic acid], indicating that these genes function exclusively in steroid degradation. PMID:12902225

  11. Structural Stability, Transitions, and Interactions within SoxYZCD-Thiosulphate from Sulfurimonas denitrificans: An In Silico Molecular Outlook for Maintaining Environmental Sulphur Cycle

    PubMed Central

    Banerjee, Arundhati

    2016-01-01

    Thiosulphate oxidation (an essential mechanism) serves to maintain the global sulphur cycle. Earlier experimental and computational studies dealt with environmental thiosulphate oxidation but none dealt with thiosulphate oxidation from deep ocean belts. Wet-laboratory experimental research shows that epsilon-proteobacteria Sulfurimonas denitrificans possess sox (sulphur-oxidizing) operon and perform thiosulphate oxidation efficiently underneath the oceans. From this specific sox operon, SoxCD complex recycles the thiosulphate-bound SoxY from SoxYZ complex to balance the environmental sulphur cycle. So, four chief proteins were variedly modeled and relevant simulated interactive structures were obtained. The final simulated tetraprotein complex (SoxYZCD) from docked SoxYZ and SoxCD complexes was disclosed to be a highly interactive one with predominant ionic residues. Free energy of folding, solvent accessibility, and conformational shifts (coil-like conformation to helices and sheets) were observed in SoxYZ complex after interacting with SoxCD. The stability of the complex (SoxYZCD) after simulation was also observed through the electrostatic surface potential values. These evaluations were rationalized via biostatistics. This aids SoxCD for recycling SoxY along with thiosulphate, which remains interconnected by four H-bonds with SoxY. Therefore, this novel exploration is endowed with the detailed molecular viewpoint for maintaining the sulphur cycle (globally) including the ocean belts. PMID:27777586

  12. The structure of RdDddP from Roseobacter denitrificans reveals that DMSP lyases in the DddP-family are metalloenzymes.

    PubMed

    Hehemann, Jan-Hendrik; Law, Adrienne; Redecke, Lars; Boraston, Alisdair B

    2014-01-01

    Marine microbes degrade dimethylsulfoniopropionate (DMSP), which is produced in large quantities by marine algae and plants, with DMSP lyases into acrylate and the gas dimethyl sulfide (DMS). Approximately 10% of the DMS vents from the sea into the atmosphere and this emission returns sulfur, which arrives in the sea through rivers and runoff, back to terrestrial systems via clouds and rain. Despite their key role in this sulfur cycle DMSP lyases are poorly understood at the molecular level. Here we report the first X-ray crystal structure of the putative DMSP lyase RdDddP from Roseobacter denitrificans, which belongs to the abundant DddP family. This structure, determined to 2.15 Å resolution, shows that RdDddP is a homodimeric metalloprotein with a binuclear center of two metal ions located 2.7 Å apart in the active site of the enzyme. Consistent with the crystallographic data, inductively coupled plasma mass spectrometry (ICP-MS) and total reflection X-ray fluorescence (TRXF) revealed the bound metal species to be primarily iron. A 3D structure guided analysis of environmental DddP lyase sequences elucidated the critical residues for metal binding are invariant, suggesting all proteins in the DddP family are metalloenzymes.

  13. The structure of RdDddP from Roseobacter denitrificans reveals that DMSP lyases in the DddP-family are metalloenzymes.

    PubMed

    Hehemann, Jan-Hendrik; Law, Adrienne; Redecke, Lars; Boraston, Alisdair B

    2014-01-01

    Marine microbes degrade dimethylsulfoniopropionate (DMSP), which is produced in large quantities by marine algae and plants, with DMSP lyases into acrylate and the gas dimethyl sulfide (DMS). Approximately 10% of the DMS vents from the sea into the atmosphere and this emission returns sulfur, which arrives in the sea through rivers and runoff, back to terrestrial systems via clouds and rain. Despite their key role in this sulfur cycle DMSP lyases are poorly understood at the molecular level. Here we report the first X-ray crystal structure of the putative DMSP lyase RdDddP from Roseobacter denitrificans, which belongs to the abundant DddP family. This structure, determined to 2.15 Å resolution, shows that RdDddP is a homodimeric metalloprotein with a binuclear center of two metal ions located 2.7 Å apart in the active site of the enzyme. Consistent with the crystallographic data, inductively coupled plasma mass spectrometry (ICP-MS) and total reflection X-ray fluorescence (TRXF) revealed the bound metal species to be primarily iron. A 3D structure guided analysis of environmental DddP lyase sequences elucidated the critical residues for metal binding are invariant, suggesting all proteins in the DddP family are metalloenzymes. PMID:25054772

  14. IncP-1β plasmids of Comamonas sp. and Delftia sp. strains isolated from a wastewater treatment plant mediate resistance to and decolorization of the triphenylmethane dye crystal violet.

    PubMed

    Stolze, Yvonne; Eikmeyer, Felix; Wibberg, Daniel; Brandis, Gerrit; Karsten, Christina; Krahn, Irene; Schneiker-Bekel, Susanne; Viehöver, Prisca; Barsch, Aiko; Keck, Matthias; Top, Eva M; Niehaus, Karsten; Schlüter, Andreas

    2012-08-01

    The application of toxic triphenylmethane dyes such as crystal violet (CV) in various industrial processes leads to large amounts of dye-contaminated sludges that need to be detoxified. Specific bacteria residing in wastewater treatment plants (WWTPs) are able to degrade triphenylmethane dyes. The objective of this work was to gain insights into the genetic background of bacterial strains capable of CV degradation. Three bacterial strains isolated from a municipal WWTP harboured IncP-1β plasmids mediating resistance to and decolorization of CV. These isolates were assigned to the genera Comamonas and Delftia. The CV-resistance plasmid pKV29 from Delftia sp. KV29 was completely sequenced. In addition, nucleotide sequences of the accessory regions involved in conferring CV resistance were determined for plasmids pKV11 and pKV36 from the other two isolates. Plasmid pKV29 contains typical IncP-1β backbone modules that are highly similar to those of previously sequenced IncP-1β plasmids that confer antibiotic resistance, degradative capabilities or mercury resistance. The accessory regions located between the conjugative transfer (tra) and mating pair formation modules (trb) of all three plasmids analysed share common modules and include a triphenylmethane reductase gene, tmr, that is responsible for decolorization of CV. Moreover, these accessory regions encode other enzymes that are dispensable for CV degradation and hence are involved in so-far-unknown metabolic pathways. Analysis of plasmid-mediated degradation of CV in Escherichia coli by ultra-high-performance liquid chromatography-electrospray ionization-quadrupole-time-of-flight MS revealed that leuco crystal violet was the first degradation product. Michler's ketone and 4-dimethylaminobenzaldehyde appeared as secondary degradation metabolites. Enzymes encoded in the E. coli chromosome seem to be responsible for cleavage of leuco crystal violet. Plasmid-mediated degradation of triphenylmethane dyes such as CV

  15. Construction of a biosensor mutant of Comamonas testosteroni for testosterone determination by cloning the EGFP gene downstream to the regulatory region of the 3,17β-HSD gene.

    PubMed

    Xiong, Guangming; Maser, Edmund

    2015-06-01

    Comamonas testosteroni (C. testosteroni) is able to catabolize a variety of steroids and polycyclic aromatic hydrocarbons. 3,17β-Hydroxysteroid dehydrogenase (3,17β-HSD) from C. testosteroni is a testosterone-inducible protein and a key enzyme in steroid degradation. 3,17β-HSD is a member of the short-chain dehydrogenase/reductase (SDR) superfamily. We found that a 2.4 kb regulatory DNA fragment upstream of the 3,17β-HSD gene (βhsd) responds to steroids and triggers βhsd gene induction. To exploit this cis-acting regulatory element for a steroid determination system, plasmids pK2.4-EGFP-4 and pBB2.4-EGFP-8 were constructed. Both plasmids contain the EGFP gene fused downstream to a 2.4 kb DNA fragment from C. testosteroni. However, whereas pK2.4-EGFP-4 could integrate into the chromosomal DNA of C. testosteroni and knock out the βhsd gene promoter, pBB2.4-EGFP-8 could replicate in C. testosteroni cells as a free plasmid DNA. After integration of pK2.4-EGFP-4 into the βhsd gene promoter, 3,17β-HSD expression could not be induced such that EGFP expression in the mutant cells was at low levels. In contrast, in C. testosteroni cells transformed with pBB2.4-EGFP-8 the expression of EGFP was induced with testosterone. Our results showed that fluorescence counts (relative fluorescence units; RFU) increased in parallel with testosterone concentrations. Of note, estradiol and cholesterol could not induce the EGFP reporter gene. In summary, this new biosensor system might be used for the specific determination of testosterone.

  16. The energy-conserving nitric-oxide-reductase system in Paracoccus denitrificans. Distinction from the nitrite reductase that catalyses synthesis of nitric oxide and evidence from trapping experiments for nitric oxide as a free intermediate during denitrification.

    PubMed

    Carr, G J; Page, M D; Ferguson, S J

    1989-02-15

    1. A Clark-type electrode that responds to nitric oxide has been used to show that cytoplasmic membrane vesicles of Paracoccus denitrificans have a nitric-oxide reductase activity. Nitrous oxide is the reaction product. NADH, succinate or isoascorbate plus 2,3,5,6-tetramethyl-1,4-phenylene diamine can act as reductants. The NADH-dependent activity is resistant to freezing of the vesicles and thus the NADH:nitric-oxide oxidoreductase activity of stored frozen vesicles provides a method for calibrating the electrode by titration of dissolved nitric oxide with NADH. The periplasmic nitrite reductase and nitrous-oxide reductase enzymes are absent from the vesicles which indicates that nitric-oxide reductase is a discrete enzyme associated with the denitrification process. This conclusion was supported by the finding that nitric-oxide reductase activity was absent from both membranes prepared from aerobically grown P. denitrificans and bovine heart submitochondrial particles. 2. The NADH: nitric-oxide oxidoreductase activity was inhibited by concentrations of antimycin or myxothiazol that were just sufficient to inhibit the cytochrome bc1 complex of the ubiquinol--cytochrome-c oxidoreductase. The activity was deduced to be proton translocating by the observations of: (a) up to 3.5-fold stimulation upon addition of an uncoupler; and (b) ATP synthesis with a P:2e ratio of 0.75. 3. Nitrite reductase of cytochrome cd1 type was highly purified from P. denitrificans in a new, high-yield, rapid two- or three-step procedure. This enzyme catalysed stoichiometric synthesis of nitric oxide. This observation, taken together with the finding that the maximum rate of NADH:nitric-oxide oxidoreductase activity catalysed by the vesicles was comparable with that of NADH:nitrate-oxidoreductase, is consistent with a role for nitric-oxide reductase in the physiological conversion of nitrate or nitrite to dinitrogen gas. 4. Intact cells of P. denitrificans also reduced nitric oxide in an

  17. Aminolaevulinate synthetase of Micrococcus denitrificans. Purification and properties of the enzyme, and the effect of growth conditions on the enzyme activity in cells.

    PubMed

    Tait, G H

    1973-02-01

    1. 5-Aminolaevulinate synthetase was detected in extracts of the non-photosynthetic bacterium Micrococcus denitrificans. 2. Activity is high in cells grown anaerobically in a defined nitrate medium, but is low in cells grown in an iron-deficient medium, and in cells grown aerobically. 3. Aminolaevulinate synthetase was purified extensively; it has a molecular weight of about 68000; apparent K(m) values for glycine, succinyl-CoA and pyridoxal phosphate are 12mm, 10mum and 11mum respectively; 2mum-haemin and 14mum-protoporphyrin inhibit by 50%. 4. The low activity of aminolaevulinate synthetase in iron-deficient cells increases on adding iron salts to cells only under conditions where protein synthesis can occur. 5. In defined nitrate medium with a high Ca(2+) concentration anaerobic growth yield is higher, but aminolaevulinate synthetase activity is lower than in cells grown in the medium with a low Ca(2+) concentration. In medium made from AnalaR constituents, growth yield is low and aminolaevulinate synthetase activity is high even in the presence of high concentrations of Ca(2+); on adding Cu(2+) (0.1mum) to the medium growth yield and aminolaevulinate synthetase activity become the same as in non-AnalaR medium. 6. Cells incubated under conditions where protein synthesis does not occur but where electron transport does, lose their aminolaevulinate synthetase activity rapidly. 7. The activities of aminolaevulinate dehydratase and succinic thiokinase do not change under any of the conditions of growth examined. 8. The possible mechanisms controlling aminolaevulinate synthetase activity and the role of this enzyme in controlling the synthesis of haem in this organism are discussed. PMID:4722442

  18. Transcriptional and metabolic regulation of denitrification in Paracoccus denitrificans allows low but significant activity of nitrous oxide reductase under oxic conditions.

    PubMed

    Qu, Zhi; Bakken, Lars R; Molstad, Lars; Frostegård, Åsa; Bergaust, Linda L

    2016-09-01

    Oxygen is known to repress denitrification at the transcriptional and metabolic levels. It has been a common notion that nitrous oxide reductase (N2 OR) is the most sensitive enzyme among the four N-oxide reductases involved in denitrification, potentially leading to increased N2 O production under suboxic or fluctuating oxygen conditions. We present detailed gas kinetics and transcription patterns from batch culture experiments with Paracoccus denitrificans, allowing in vivo estimation of e(-) -flow to O2 and N2 O under various O2 regimes. Transcription of nosZ took place concomitantly with that of narG under suboxic conditions, whereas transcription of nirS and norB was inhibited until O2 levels approached 0 μM in the liquid. Catalytically functional N2 OR was synthesized and active in aerobically raised cells transferred to vials with 7 vol% O2 in headspace, but N2 O reduction rates were 10 times higher when anaerobic pre-cultures were subjected to the same conditions. Upon oxygen exposure, there was an incomplete and transient inactivation of N2 OR that could be ascribed to its lower ability to compete for electrons compared with terminal oxidases. The demonstrated reduction of N2 O at high O2 partial pressure and low N2 O concentrations by a bacterium not known as a typical aerobic denitrifier may provide one clue to the understanding of why some soils appear to act as sinks rather than sources for atmospheric N2 O. PMID:26568281

  19. Cloning and Characterization of a Gene Cluster Involved in Cyclopentanol Metabolism in Comamonas sp. Strain NCIMB 9872 and Biotransformations Effected by Escherichia coli-Expressed Cyclopentanone 1,2-Monooxygenase†

    PubMed Central

    Iwaki, Hiroaki; Hasegawa, Yoshie; Wang, Shaozhao; Kayser, Margaret M.; Lau, Peter C. K.

    2002-01-01

    Cyclopentanone 1,2-monooxygenase, a flavoprotein produced by Pseudomonas sp. strain NCIMB 9872 upon induction by cyclopentanol or cyclopentanone (M. Griffin and P. W. Trudgill, Biochem. J. 129:595-603, 1972), has been utilized as a biocatalyst in Baeyer-Villiger oxidations. To further explore this biocatalytic potential and to discover new genes, we have cloned and sequenced a 16-kb chromosomal locus of strain 9872 that is herein reclassified as belonging to the genus Comamonas. Sequence analysis revealed a cluster of genes and six potential open reading frames designated and grouped in at least four possible transcriptional units as (orf11-orf10-orf9)-(cpnE-cpnD-orf6-cpnC)-(cpnR-cpnB-cpnA)-(orf3-orf4 [partial 3′ end]). The cpnABCDE genes encode enzymes for the five-step conversion of cyclopentanol to glutaric acid catalyzed by cyclopentanol dehydrogenase, cyclopentanone 1,2-monooxygenase, a ring-opening 5-valerolactone hydrolase, 5-hydroxyvalerate dehydrogenase, and 5-oxovalerate dehydrogenase, respectively. Inactivation of cpnB by using a lacZ-Kmr cassette resulted in a strain that was not capable of growth on cyclopentanol or cyclopentanone as a sole carbon and energy source. The presence of σ54-dependent regulatory elements in front of the divergently transcribed cpnB and cpnC genes supports the notion that cpnR is a regulatory gene of the NtrC type. Knowledge of the nucleotide sequence of the cpn genes was used to construct isopropyl-β-thio-d-galactoside-inducible clones of Escherichia coli cells that overproduce the five enzymes of the cpn pathway. The substrate specificities of CpnA and CpnB were studied in particular to evaluate the potential of these enzymes and establish the latter recombinant strain as a bioreagent for Baeyer-Villiger oxidations. Although frequently nonenantioselective, cyclopentanone 1,2-monooxygenase was found to exhibit a broader substrate range than the related cyclohexanone 1,2-monooxygenase from Acinetobacter sp. strain NCIMB

  20. Crystal structures of cytochrome c(L) and methanol dehydrogenase from Hyphomicrobium denitrificans: structural and mechanistic insights into interactions between the two proteins.

    PubMed

    Nojiri, Masaki; Hira, Daisuke; Yamaguchi, Kazuya; Okajima, Toshihide; Tanizawa, Katsuyuki; Suzuki, Shinnichiro

    2006-03-21

    Methanol dehydrogenase (Hd-MDH) and its physiological electron acceptor, cytochrome c(L) (Hd-Cyt c(L)), isolated from a methylotrophic denitrifying bacterium, Hyphomicrobium denitrificans A3151, have been kinetically and structurally characterized; the X-ray structures of Hd-MDH and Hd-Cyt c(L) have been determined using molecular replacement at 2.5 and 2.0 A resolution, respectively. To explain the mechanism for electron transfer between these proteins, the dependence of MDH activity on the concentration of Hd-Cyt c(L) has been investigated at pH 4.5-7.0. The Michaelis constant for Hd-Cyt c(L) shows the smallest value (approximately 0.3 microM) at pH 5.5. The pseudo-first-order rate constant (k(obs)) of the reduction of Hd-Cyt c(L) exhibits a hyperbolic concentration dependence of Hd-MDH at pH 5.5, although k(obs) linearly increases at pH 6.5. These findings indicate formation of a transient complex between these proteins during an electron transfer event. Hd-MDH (148 kDa) is a large tetrameric protein with an alpha(2)beta(2) subunit composition, showing a high degree of structural similarity with other MDHs. Hd-Cyt c(L) (19 kDa) exhibiting the alpha-band at 550.7 nm has a unique C-terminal region involving a disulfide bond between Cys47 and Cys165. Moreover, there is a pair of Hd-Cyt c(L) monomers related with a pseudo-2-fold axis of symmetry in the asymmetric unit, and the two monomers tightly interact with each other through three hydrogen bonds. This configuration is the first example in the studies of cytochrome c as the physiological electron acceptor for MDH. The docking simulation between the coupled Hd-Cyt c(L) molecules and the heterotetrameric Hd-MDH molecule has been carried out. PMID:16533029

  1. FnrP and NNR of Paracoccus denitrificans are both members of the FNR family of transcriptional activators but have distinct roles in respiratory adaptation in response to oxygen limitation.

    PubMed

    Van Spanning, R J; De Boer, A P; Reijnders, W N; Westerhoff, H V; Stouthamer, A H; Van Der Oost, J

    1997-03-01

    The Paracoccus denitrificans fnrP gene encoding a homologue of the Escherichia coli FNR protein was localized upstream of the gene cluster that encodes the high-affinity cbb3-type oxidase. FnrP harbours the invariant cysteine residues that are supposed to be the ligands of the redox-sensitive [4Fe-4S] cluster in FNR. NNR, another FNR-like transcriptional regulator in P. denitrificans, does not. Analysis of FnrP and NNR single and double mutants revealed that the two regulators each exert exclusive control on the expression of a discrete set of target genes. In FnrP mutants, the expression of cytochrome c peroxidase was blocked, that of membrane-bound nitrate reductase and the cbb3-type oxidase was significantly reduced, whilst the activity of the bb3-type quinol oxidase was increased. The amounts of the nitrite and nitric oxide reductases in these FnrP mutants were the same as in the wild type. NNR mutants, on the other hand, were disturbed exclusively in the concentrations of nitrite reductase and nitric oxide reductase. An FnrP.NNR double mutant combined the phenotypes of the single mutant strains. In all three mutants, the concentrations and/or activities of the aa3-type oxidase, cytochrome C550, cytochrome C552, and nitrous oxide reductase equalled those in the wild type. As the FNR boxes in front of the FnrP- and NNR-regulated genes are highly similar to or even identical to each other, the absence of cross-talk between the regulation by FnrP and NNR implies that as yet unidentified factors are important in the control. It is proposed that the redox state of an intracellular redox couple other than the oxygen/water couple is one of the factors that modulates the activity of FnrP. PMID:9076727

  2. Phylogenetic origins of the plant mitochondrion based on a comparative analysis of 5S ribosomal RNA sequences

    NASA Technical Reports Server (NTRS)

    Villanueva, E.; Delihas, N.; Luehrsen, K. R.; Fox, G. E.; Gibson, J.

    1985-01-01

    The complete nucleotide sequences of 5S ribosomal RNAs from Rhodocyclus gelatinosa, Rhodobacter sphaeroides, and Pseudomonas cepacia were determined. Comparisons of these 5S RNA sequences show that rather than being phylogenetically related to one another, the two photosynthetic bacterial 5S RNAs share more sequence and signature homology with the RNAs of two nonphotosynthetic strains. Rhodobacter sphaeroides is specifically related to Paracoccus denitrificans and Rc. gelatinosa is related to Ps. cepacia. These results support earlier 16S ribosomal RNA studies and add two important groups to the 5S RNA data base. Unique 5S RNA structural features previously found in P. denitrificans are present also in the 5S RNA of Rb. sphaeroides; these provide the basis for subdivisional signatures. The immediate consequence of obtaining these new sequences is that it is possible to clarify the phylogenetic origins of the plant mitochondrion. In particular, a close phylogenetic relationship is found between the plant mitochondria and members of the alpha subdivision of the purple photosynthetic bacteria, namely, Rb. sphaeroides, P. denitrificans, and Rhodospirillum rubrum.

  3. Identification of 9α-Hydroxy-17-Oxo-1,2,3,4,10,19-Hexanorandrostan-5-Oic Acid in Steroid Degradation by Comamonas testosteroni TA441 and Its Conversion to the Corresponding 6-En-5-Oyl Coenzyme A (CoA) Involving Open Reading Frame 28 (ORF28)- and ORF30-Encoded Acyl-CoA Dehydrogenases

    PubMed Central

    Hayashi, Toshiaki; Koshino, Hiroyuki; Malon, Michal; Hirota, Hiroshi; Kudo, Toshiaki

    2014-01-01

    Comamonas testosteroni TA441 degrades steroids via aromatization and meta-cleavage of the A ring, followed by hydrolysis, and produces 9,17-dioxo-1,2,3,4,10,19-hexanorandrostan-5-oic acid as an intermediate compound. Herein, we identify a new intermediate compound, 9α-hydroxy-17-oxo-1,2,3,4,10,19-hexanorandrostan-5-oic acid. Open reading frame 28 (ORF28)- and ORF30-encoded acyl coenzyme A (acyl-CoA) dehydrogenase was shown to convert the CoA ester of 9α-hydroxy-17-oxo-1,2,3,4,10,19-hexanorandrostan-5-oic acid to the CoA ester of 9α-hydroxy-17-oxo-1,2,3,4,10,19-hexanorandrost-6-en-5-oic acid. A homology search of the deduced amino acid sequences suggested that the ORF30-encoded protein is a member of the acyl-CoA dehydrogenase_fadE6_17_26 family, whereas the deduced amino acid sequence of ORF28 showed no significant similarity to specific acyl-CoA dehydrogenase family proteins. Possible steroid degradation gene clusters similar to the cluster of TA441 appear in bacterial genome analysis data. In these clusters, ORFs similar to ORFs 28 and 30 are often found side by side and ordered in the same manner as ORFs 28 and 30. PMID:25092028

  4. The designated type strain of Pseudomonas halophila Fendrich 1989 is DSM 3051, the designated type strain of Halovibrio variabilis Fendrich 1989 is DSM 3050, the new name Halomonas utahensis (Fendrich 1989) Sorokin and Tindall 2006 is created for the species represented by DSM 3051 when treated as a member of the genus Halomonas, the combination Halomonas variabilis (Fendrich 1989) Dobson and Franzmann 1996 is rejected, and the combination Halovibrio denitrificans Sorokin et al. 2006 is validly published with an emendation of the description of the genus Halovibrio Fendrich 1989 emend. Sorokin et al. 2006. Opinion 93. Judicial Commission of the International Committee on Systematics of Prokaryotes.

    PubMed

    Tindall, B J

    2014-10-01

    The Judicial Commission affirms that, according to information presented to it, the designated type strain of Pseudomonas halophila Fendrich 1989 is DSM 3051 (replacing DSM 3050) and the designated type strain of Halovibrio variabilis Fendrich 1989 is DSM 3050 (replacing DSM 3051). A new name, Halomonas utahensis (Fendrich 1989) Sorokin and Tindall 2006 nom. nov., is created for the species represented by DSM 3051 when treated as a member of the genus Halomonas, because the combination Halomonas halophila (Quesada et al. 1984) Dobson and Franzmann 1996 has priority based on the fact that the epithet halophila in the combination Halomonas halophila (Quesada et al. 1984) Dobson and Franzmann 1996 ( BASOYNM: Deleya halophila Quesada et al. 1984) has priority over the epithet halophila should the taxon Pseudomonas halophila Fendrich 1989 be treated as a member of the genus Halomonas. The combination Halomonas variabilis (Fendrich 1989) Dobson and Franzmann 1996 is rejected. The combination Halovibrio denitrificans Sorokin et al. 2006 is validly published with an emendation of the description of the genus Halovibrio Fendrich 1989 emend. Sorokin et al. 2006.

  5. Pyrite-based autotrophic denitrification for remediation of nitrate contaminated groundwater.

    PubMed

    Pu, Jiaoyang; Feng, Chuanping; Liu, Ying; Li, Rui; Kong, Zhe; Chen, Nan; Tong, Shuang; Hao, Chunbo; Liu, Ye

    2014-12-01

    In this study, pyrite-based denitrification using untreated pyrite (UP) and acid-pretreated pyrite (AP) was evaluated as an alternative to elemental sulfur based denitrification. Pyrite-based denitrification resulted in a favorable nitrate removal rate constant (0.95 d(-1)), sulfate production of 388.00 mg/L, and a stable pH. The pretreatment of pyrite with acid led to a further increase in the nitrate removal rate constant (1.03 d(-1)) and reduction in initial sulfate concentration (224.25±7.50 mg/L). By analyzing the microbial community structure using Denaturing Gradient Gel Electrophoresis, it was confirmed that Sulfurimonas denitrificans (S. denitrificans) could utilize pyrite as an electron donor. A stable pH was observed over the entire experimental period, indicating that the use of a pH buffer reagent would not be necessary for pyrite-based denitrification. Therefore, pyrite could effectively replace elemental sulfur as an electron donor in autotrophic denitrification for nitrate-contaminated groundwater remediation.

  6. INDISIM-Paracoccus, an individual-based and thermodynamic model for a denitrifying bacterium.

    PubMed

    Araujo Granda, Pablo; Gras, Anna; Ginovart, Marta; Moulton, Vincent

    2016-08-21

    We have developed an individual-based model for denitrifying bacteria. The model, called INDISIM-Paracoccus, embeds a thermodynamic model for bacterial yield prediction inside the individual-based model INDISIM, and is designed to simulate the bacterial cell population behavior and the product dynamics within the culture. The INDISIM-Paracoccus model assumes a culture medium containing succinate as a carbon source, ammonium as a nitrogen source and various electron acceptors such as oxygen, nitrate, nitrite, nitric oxide and nitrous oxide to simulate in continuous or batch culture the different nutrient-dependent cell growth kinetics of the bacterium Paracoccus denitrificans. The individuals in the model represent microbes and the individual-based model INDISIM gives the behavior-rules that they use for their nutrient uptake and reproduction cycle. Three previously described metabolic pathways for P. denitrificans were selected and translated into balanced chemical equations using a thermodynamic model. These stoichiometric reactions are an intracellular model for the individual behavior-rules for metabolic maintenance and biomass synthesis and result in the release of different nitrogen oxides to the medium. The model was implemented using the NetLogo platform and it provides an interactive tool to investigate the different steps of denitrification carried out by a denitrifying bacterium. The simulator can be obtained from the authors on request. PMID:27179457

  7. Phylogenetic relationships among members of the Comamonadaceae, and description of Delftia acidovorans (den Dooren de Jong 1926 and Tamaoka et al. 1987) gen. nov., comb. nov.

    PubMed

    Wen, A; Fegan, M; Hayward, C; Chakraborty, S; Sly, L I

    1999-04-01

    The phylogenetic relationships among members of the family Comamonadaceae and several unclassified strains were studied by direct sequencing of their PCR-amplified 16S rRNA genes. Based on the 16S rRNA gene sequence analysis, members of the family formed a coherent group. The closest relatives are species of the Rubrivivax sub-group: Leptothrix discophora, Ideonella dechloratans and Rubrivivax gelatinosus. The genus Hydrogenophaga formed two subclusters, as did the species of Acidovorax, whereas the five species of the genus [Aquaspirillum] were polyphyletic. Comamonas acidovorans was phylogenetically distant from the type species of Comamonas, Comamonas terrigena. On the basis of this work and previous studies, Comamonas acidovorans is removed from the genus Comamonas and renamed as Delftia acidovorans gen. nov., comb. nov. Descriptions of the new genus Delftia and of the type species Delftia acidovorans, for which the type strain is ATCC 15668T, are presented. PMID:10319477

  8. Remediation of oil-based drill cuttings through a biosurfactant-based washing followed by a biodegradation treatment.

    PubMed

    Yan, Ping; Lu, Mang; Guan, Yueming; Zhang, Weimu; Zhang, Zhongzhi

    2011-11-01

    In this study, oil-based drill cuttings were washed by a rhamnolipid solution and then subjected to bioremediation in stainless steel boxes using sawdust as bulking agent. A mixed bacterial culture, mainly containing Pseudomonas, Acinetobacter, Alcaligenes, Agrobacterium, and Comamonas, was used as inoculums. Approximately 83% of organics were removed after washing under optimal conditions (liquid/solid ratio, 3:1; washing time, 20 min; stirring speed, 200 rpm; rhamnolipid concentration, 360 mg/L; temperature, 60 °C), and the total petroleum hydrocarbon concentration of the cuttings dropped from 85,000 to 12,600 mg/kg. In the bioremediation stage, concentrations of saturated and aromatic hydrocarbons decreased to 2140 and 1290 mg/kg, respectively, after 120 days. Ultrahigh-resolution mass spectrometry demonstrated that oxygen- and nitrogen-containing compounds had undergone biodegradation. The results of this study indicate that this two-stage remedial system can reduce treatment time and increase treatment efficiency as compared with a single bioremediation or washing treatment.

  9. Evaluation of the Catalytic Contribution from a Positioned General Base in Ketosteroid Isomerase.

    PubMed

    Lamba, Vandana; Yabukarski, Filip; Pinney, Margaux; Herschlag, Daniel

    2016-08-10

    Proton transfer reactions are ubiquitous in enzymes and utilize active site residues as general acids and bases. Crystal structures and site-directed mutagenesis are routinely used to identify these residues, but assessment of their catalytic contribution remains a major challenge. In principle, effective molarity measurements, in which exogenous acids/bases rescue the reaction in mutants lacking these residues, can estimate these catalytic contributions. However, these exogenous moieties can be restricted in reactivity by steric hindrance or enhanced by binding interactions with nearby residues, thereby resulting in over- or underestimation of the catalytic contribution, respectively. With these challenges in mind, we investigated the catalytic contribution of an aspartate general base in ketosteroid isomerase (KSI) by exogenous rescue. In addition to removing the general base, we systematically mutated nearby residues and probed each mutant with a series of carboxylate bases of similar pKa but varying size. Our results underscore the need for extensive and multifaceted variation to assess and minimize steric and positioning effects and determine effective molarities that estimate catalytic contributions. We obtained consensus effective molarities of ∼5 × 10(4) M for KSI from Comamonas testosteroni (tKSI) and ∼10(3) M for KSI from Pseudomonas putida (pKSI). An X-ray crystal structure of a tKSI general base mutant showed no additional structural rearrangements, and double mutant cycles revealed similar contributions from an oxyanion hole mutation in the wild-type and base-rescued reactions, providing no indication of mutational effects extending beyond the general base site. Thus, the high effective molarities suggest a large catalytic contribution associated with the general base. A significant portion of this effect presumably arises from positioning of the base, but its large magnitude suggests the involvement of additional catalytic mechanisms as well

  10. Evaluation of the Catalytic Contribution from a Positioned General Base in Ketosteroid Isomerase.

    PubMed

    Lamba, Vandana; Yabukarski, Filip; Pinney, Margaux; Herschlag, Daniel

    2016-08-10

    Proton transfer reactions are ubiquitous in enzymes and utilize active site residues as general acids and bases. Crystal structures and site-directed mutagenesis are routinely used to identify these residues, but assessment of their catalytic contribution remains a major challenge. In principle, effective molarity measurements, in which exogenous acids/bases rescue the reaction in mutants lacking these residues, can estimate these catalytic contributions. However, these exogenous moieties can be restricted in reactivity by steric hindrance or enhanced by binding interactions with nearby residues, thereby resulting in over- or underestimation of the catalytic contribution, respectively. With these challenges in mind, we investigated the catalytic contribution of an aspartate general base in ketosteroid isomerase (KSI) by exogenous rescue. In addition to removing the general base, we systematically mutated nearby residues and probed each mutant with a series of carboxylate bases of similar pKa but varying size. Our results underscore the need for extensive and multifaceted variation to assess and minimize steric and positioning effects and determine effective molarities that estimate catalytic contributions. We obtained consensus effective molarities of ∼5 × 10(4) M for KSI from Comamonas testosteroni (tKSI) and ∼10(3) M for KSI from Pseudomonas putida (pKSI). An X-ray crystal structure of a tKSI general base mutant showed no additional structural rearrangements, and double mutant cycles revealed similar contributions from an oxyanion hole mutation in the wild-type and base-rescued reactions, providing no indication of mutational effects extending beyond the general base site. Thus, the high effective molarities suggest a large catalytic contribution associated with the general base. A significant portion of this effect presumably arises from positioning of the base, but its large magnitude suggests the involvement of additional catalytic mechanisms as well.

  11. First microbiota assessments of children's paddling pool waters evaluated using 16S rRNA gene-based metagenome analysis.

    PubMed

    Sawabe, Toko; Suda, Wataru; Ohshima, Kenshiro; Hattori, Masahira; Sawabe, Tomoo

    2016-01-01

    Insufficient chloric sterilization of children's paddling pool waters increases the risk of diarrheal illness. Therefore, we investigated the microbiota changes after children use pools. First, we applied 16S rRNA gene-based metagenome analysis to understand the dynamics of microbiota in pool water, especially with respect to the bio-contamination by potential pathogens. Proteobacteria were major taxa detected in every pool water sample after children spent time in the pool. In more detail, Gammaproteobacteria comprised the dominant class, which was followed by Betaproteobacteria. Five phyla, Bacteroidetes, Firmicutes, Actinobacteria and Deinococcus-Thermus phyla were minor groups. The pool water microbiota are likely to be a consortium of intestinal and skin microbiota from humans. Interestingly, the ratio of Gammaproteobacteria and Betaproteobacteria differed according to the age of the children who used the pool, which means the pool water was additionally contaminated by soil microbiota as a result of the children's behavior. Furthermore, potential pathogens, such as Campylobacter spp., Comamonas testosteroni and Burkholderia pseudomallei, were also found. Considering the standard plate counts, the abundances of these human pathogens are unlikely to be a sufficiently infectious dose. We suggest the importance of sanitary measures in paddling pool waters to reduce bio-contamination from both humans and the environment. PMID:26671497

  12. First microbiota assessments of children's paddling pool waters evaluated using 16S rRNA gene-based metagenome analysis.

    PubMed

    Sawabe, Toko; Suda, Wataru; Ohshima, Kenshiro; Hattori, Masahira; Sawabe, Tomoo

    2016-01-01

    Insufficient chloric sterilization of children's paddling pool waters increases the risk of diarrheal illness. Therefore, we investigated the microbiota changes after children use pools. First, we applied 16S rRNA gene-based metagenome analysis to understand the dynamics of microbiota in pool water, especially with respect to the bio-contamination by potential pathogens. Proteobacteria were major taxa detected in every pool water sample after children spent time in the pool. In more detail, Gammaproteobacteria comprised the dominant class, which was followed by Betaproteobacteria. Five phyla, Bacteroidetes, Firmicutes, Actinobacteria and Deinococcus-Thermus phyla were minor groups. The pool water microbiota are likely to be a consortium of intestinal and skin microbiota from humans. Interestingly, the ratio of Gammaproteobacteria and Betaproteobacteria differed according to the age of the children who used the pool, which means the pool water was additionally contaminated by soil microbiota as a result of the children's behavior. Furthermore, potential pathogens, such as Campylobacter spp., Comamonas testosteroni and Burkholderia pseudomallei, were also found. Considering the standard plate counts, the abundances of these human pathogens are unlikely to be a sufficiently infectious dose. We suggest the importance of sanitary measures in paddling pool waters to reduce bio-contamination from both humans and the environment.

  13. Removal of methyl acrylate by ceramic-packed biotrickling filter and their response to bacterial community.

    PubMed

    Wu, Hao; Yin, Zhenhao; Quan, Yue; Fang, Yingyu; Yin, Chengri

    2016-06-01

    Methyl acrylate is a widely used raw chemical materials and it is toxic in humans. In order to treat the methyl acrylate waste gas, a 3-layer BTF packed with ceramic particles and immobilized with activated sludge was set up. The BTF exhibited excellent removal efficiency that no methyl acrylate could be detected when EBRT was larger than 266s and inlet concentration was lower than 0.19g/m(3). The 1st layer performed the best at fixed inlet concentration of 0.42g/m(3). PCR combined with DGGE was performed to detect the differences in different layers of the BTF. Phylum Proteobacteria (e.g. α-, β-, γ-, δ-) was predominantly represented in the bacterial community, followed by Actinobacteria and Firmicutes. Desulfovibrio gigas, Variovorax paradoxus, Dokdonella koreensis, Pseudoxanthomonas suwonensis, Azorhizobium caulinodans, Hyphomicrobium denitrificans, Hyphomicrobium sp. and Comamonas testosteroni formed the bacteria community to treat methyl acrylate waste gas in the BTF. PMID:26970927

  14. [Study on sulfur-based autotrophic denitrification with different electron donors].

    PubMed

    Yuan, Ying; Zhou, Wei-li; Wang, Hui; He, Sheng-bing

    2013-05-01

    Sulphur-based autotrophic denitrification was applied to treat the low concentration nitrate-contaminated water. Different electron donors, namely, elemental sulfur, sulfide and thiosulfate, were used in three continuous reactors to compare the denitrification performance. When treating the low concentration nitrate-contaminated water (13 mg x L(-1)), the thiosulfate system showed the best performance and the sulfide system was the worst. The thiosulfate system was less sensitive to low temperature than the other two. At temperatures higher than 20 degrees C, the sulfur system was greatly influenced by mass transfer efficiency and HRT. It removed 81% of nitrate and 79% of TN when HRT was no less than 2 h, but could only achieve a low nitrate and TN removal rate of 47% and 51% when HRT was shorter than 2 h. No obvious nitrite accumulation was observed and the average effluent nitrite was 0.53 mg x L(-1). The sulfide system could only remove 47% of nitrate and 41% of TN with 0.29 mg x L(-1) nitrite in the effluent at the HRT of 4 h. Meanwhile, the thiosulfate system had a high removal rate of 99% for nitrate and 90% for TN, with a low content of effluent nitrite of 0.080 mg x L(-1), and the HRT could be shortened to 0.5 h. The molecular biological analysis showed that different bacteria predominated in the three reactors, and that Thiobacillus denitrificans existed abundantly in the sulfur system, while the functional bacteria in the sulfide and thiosulfate systems could not be identified yet. A new species of sulfur-based autotrophic denitrification bacterium may have been found.

  15. Energy coupling to nitrate uptake into the denitrifying cells of Paracoccus denitrificans.

    PubMed

    Kucera, Igor

    2005-09-01

    This study deals with the effects of the agents that dissipate the individual components of the proton motive force (short-chain fatty acids, nigericin, and valinomycin) upon the methyl viologen-coupled nitrate reductase activity in intact cells. Substitution of butyrate or acetate for chloride in Tris-buffered assay media resulted in a marked inhibition at pH 7. In a Tris--chloride buffer of neutral pH, the reaction was almost fully inhibitable by nigericin. Alkalinisation increased the IC(50) value for nigericin and decreased the maximal inhibition attained. Both types of inhibitions could be reversed by the permeabilisation of cells or by the addition of nitrite, and that caused by nigericin disappeared at high extracellular concentrations of potassium. These data indicate that nitrate transport step relies heavily on the pH gradient at neutral pH. Since the affinity of cells for nitrate was strongly diminished by imposing an inside-positive potassium (or lithium) diffusion potential at alkaline external pH, a potential dependent step may be of significance in the transporter cycle under these conditions. Experiments with sodium-depleted media provided no hints for Na(+) as a possible H(+) substitute. PMID:16112075

  16. Microbial recovery of metals from spent coal liquefaction catalysts. [Thiobacillus denitrificans, Sulfolobus

    SciTech Connect

    Sperl, P.L.; Sperl, G.T.

    1991-01-01

    This project was initiated on October 1, 1989, for the purpose of recovering metals from spent coal liquefaction catalysts. The catalyst is a Ni-Mo catalyst supported on alumina (Shell 324) as is used in a pilot scale coal liquefaction facility at Wilsonville, Alabama. This plant is run and operated by Southern Clean Fuels. A large sample of spent catalyst from this facility has been obtained. The object of the contract is to treat the spent catalysts with microorganisms, especially Thiobacillus ferrooxidans, but also other Thiobacillus sp. and possibly Sulfolobus, and other potentially useful microorganisms to leach and remove the metals (Ni and Mo) form the spent catalysts into a form which can be readily recovered by conventional techniques.

  17. A nitrite biosensor based on co-immobilization of nitrite reductase and viologen-modified chitosan on a glassy carbon electrode.

    PubMed

    Quan, De; Shin, Woonsup

    2010-01-01

    An electrochemical nitrite biosensor based on co-immobilization of copper-containing nitrite reductase (Cu-NiR, from Rhodopseudomonas sphaeroides forma sp. denitrificans) and viologen-modified chitosan (CHIT-V) on a glassy carbon electrode (GCE) is presented. Electron transfer (ET) between a conventional GCE and immobilized Cu-NiR was mediated by the co-immobilized CHIT-V. Redox-active viologen was covalently linked to a chitosan backbone, and the thus produced CHIT-V was co-immobilized with Cu-NiR on the GCE surface by drop-coating of hydrophilic polyurethane (HPU). The electrode responded to nitrite with a limit of detection (LOD) of 40 nM (S/N = 3). The sensitivity, linear response range, and response time (t(90%)) were 14.9 nA/μM, 0.04-11 μM (r(2) = 0.999) and 15 s, respectively. The corresponding Lineweaver-Burk plot showed that the apparent Michaelis-Menten constant (K(M) (app)) was 65 μM. Storage stability of the biosensor (retaining 80% of initial activity) was 65 days under ambient air and room temperature storage conditions. Reproducibility of the sensor showed a relative standard deviation (RSD) of 2.8% (n = 5) for detection of 1 μM of nitrite. An interference study showed that anions commonly found in water samples such as chlorate, chloride, sulfate and sulfite did not interfere with the nitrite detection. However, nitrate interfered with a relative sensitivity of 64% and this interference effect was due to the intrinsic character of the NiR employed in this study. PMID:22219710

  18. Development and Use of Integrated Microarray-Based Genomic Technologies for Assessing Microbial Community Composition and Dynamics

    SciTech Connect

    Zhou, J.; Wu, L.; Gentry, T.; Schadt, C.; He, Z.; Li, X.

    2006-04-05

    To effectively monitor microbial populations involved in various important processes, a 50-mer-based oligonucleotide microarray was developed based on known genes and pathways involved in: biodegradation, metal resistance and reduction, denitrification, nitrification, nitrogen fixation, methane oxidation, methanogenesis, carbon polymer decomposition, and sulfate reduction. This array contains approximately 2000 unique and group-specific probes with <85% similarity to their non-target sequences. Based on artificial probes, our results showed that at hybridization conditions of 50 C and 50% formamide, the 50-mer microarray hybridization can differentiate sequences having <88% similarity. Specificity tests with representative pure cultures indicated that the designed probes on the arrays appeared to be specific to their corresponding target genes. Detection limits were about 5-10ng genomic DNA in the absence of background DNA, and 50-100ng ({approx}1.3{sup o} 10{sup 7} cells) in the presence background DNA. Strong linear relationships between signal intensity and target DNA and RNA concentration were observed (r{sup 2} = 0.95-0.99). Application of this microarray to naphthalene-amended enrichments and soil microcosms demonstrated that composition of the microflora varied depending on incubation conditions. While the naphthalene-degrading genes from Rhodococcus-type microorganisms were dominant in enrichments, the genes involved in naphthalene degradation from Gram-negative microorganisms such as Ralstonia, Comamonas, and Burkholderia were most abundant in the soil microcosms (as well as those for polyaromatic hydrocarbon and nitrotoluene degradation). Although naphthalene degradation is widely known and studied in Pseudomonas, Pseudomonas genes were not detected in either system. Real-time PCR analysis of 4 representative genes was consistent with microarray-based quantification (r{sup 2} = 0.95). Currently, we are also applying this microarray to the study of several

  19. Development and Use of Integrated Microarray-Based Genomic Technologies for Assessing Microbial Community Composition and Dynamics

    SciTech Connect

    J. Zhou; S.-K. Rhee; C. Schadt; T. Gentry; Z. He; X. Li; X. Liu; J. Liebich; S.C. Chong; L. Wu

    2004-03-17

    To effectively monitor microbial populations involved in various important processes, a 50-mer-based oligonucleotide microarray was developed based on known genes and pathways involved in: biodegradation, metal resistance and reduction, denitrification, nitrification, nitrogen fixation, methane oxidation, methanogenesis, carbon polymer decomposition, and sulfate reduction. This array contains approximately 2000 unique and group-specific probes with <85% similarity to their non-target sequences. Based on artificial probes, our results showed that at hybridization conditions of 50 C and 50% formamide, the 50-mer microarray hybridization can differentiate sequences having <88% similarity. Specificity tests with representative pure cultures indicated that the designed probes on the arrays appeared to be specific to their corresponding target genes. Detection limits were about 5-10ng genomic DNA in the absence of background DNA, and 50-100ng ({approx}1.3{sup o} 10{sup 7} cells) in the presence background DNA. Strong linear relationships between signal intensity and target DNA and RNA concentration were observed (r{sup 2} = 0.95-0.99). Application of this microarray to naphthalene-amended enrichments and soil microcosms demonstrated that composition of the microflora varied depending on incubation conditions. While the naphthalene-degrading genes from Rhodococcus-type microorganisms were dominant in enrichments, the genes involved in naphthalene degradation from Gram-negative microorganisms such as Ralstonia, Comamonas, and Burkholderia were most abundant in the soil microcosms (as well as those for polyaromatic hydrocarbon and nitrotoluene degradation). Although naphthalene degradation is widely known and studied in Pseudomonas, Pseudomonas genes were not detected in either system. Real-time PCR analysis of 4 representative genes was consistent with microarray-based quantification (r{sup 2} = 0.95). Currently, we are also applying this microarray to the study of several

  20. Evaluation of pyrrolidonyl arylamidase for the identification of nonfermenting Gram-negative rods.

    PubMed

    Bombicino, Karina A; Almuzara, Marisa N; Famiglietti, Angela M R; Vay, Carlos

    2007-01-01

    To evaluate the activity of pyrrolidonyl arylamidase (PYR) for the differentiation and identification of nonfermenting gram negative rods (NFGNR), 293 isolates were tested. A 24 h culture of each test organism was prepared. From this a 108-109 cfu/mL suspension was added to 0.25 mL of sterile physiologic solution. A PYR disk was then added and the test was incubated for 30 minutes at 35-37 degrees C, at environmental atmosphere. Reading was done by adding 1 drop of cinnamaldehyde reagent. Strains of Acinetobacter baumannii, Acinetobacter haemolyticus, Alcaligenes faecalis, Bergeyella zoohelcum, Bordetella bronchiseptica, Bordetella hinzii, Brevundimonas diminuta, Brevundimonas vesicularis, Brucella ovis, Brucella spp., Brucella suis, Burkholderia cepacia complex, Moraxella catarrhalis, Moraxella lacunata, Moraxella nonliquefaciens, Moraxella osloensis, Oligella ureolytica, Pseudomonas alcaligenes, Pseudomonas mendocina, Pseudomonas pseudoalcaligenes, Pseudomonas putida, Pseudomonas stutzeri, Pseudomonas Vb3, Psychrobacter phenylpyruvicus, and Stenotrophomonas maltophilia were PYR negative. On the other hand Achromobacter piechaudii, Achromobacter denitrificans, Achromobacter xylosoxidans, Burkholderia gladioli, Chryseobacterium gleum-indologenes, Comamonas testosroni, Cupriavidus pauculus, Delftia acidovorans, Elizabethkingia meningoseptica, Myroides spp., Ochrobactrum anthropi, Pseudomonas oryzihabitans, Ralstonia pickettii, Rhizobium radiobacter, Shewanella spp., Sphingobacterium multivorum, Sphingobacterium spiritivorum, and Weeksella virosa were PYR positive. Finally, Acinetobacter lwoffii, Pseudomonas aeruginosa, Pseudomonas fluorescens, Roseomonas spp., and Sphingomonas paucimobilis-parapaucimobilis were PYR variable. PYR testing should be considered as a useful tool to facilitate the identification of NFGNR.

  1. Evaluation of pyrrolidonyl arylamidase for the identification of nonfermenting Gram-negative rods.

    PubMed

    Bombicino, Karina A; Almuzara, Marisa N; Famiglietti, Angela M R; Vay, Carlos

    2007-01-01

    To evaluate the activity of pyrrolidonyl arylamidase (PYR) for the differentiation and identification of nonfermenting gram negative rods (NFGNR), 293 isolates were tested. A 24 h culture of each test organism was prepared. From this a 108-109 cfu/mL suspension was added to 0.25 mL of sterile physiologic solution. A PYR disk was then added and the test was incubated for 30 minutes at 35-37 degrees C, at environmental atmosphere. Reading was done by adding 1 drop of cinnamaldehyde reagent. Strains of Acinetobacter baumannii, Acinetobacter haemolyticus, Alcaligenes faecalis, Bergeyella zoohelcum, Bordetella bronchiseptica, Bordetella hinzii, Brevundimonas diminuta, Brevundimonas vesicularis, Brucella ovis, Brucella spp., Brucella suis, Burkholderia cepacia complex, Moraxella catarrhalis, Moraxella lacunata, Moraxella nonliquefaciens, Moraxella osloensis, Oligella ureolytica, Pseudomonas alcaligenes, Pseudomonas mendocina, Pseudomonas pseudoalcaligenes, Pseudomonas putida, Pseudomonas stutzeri, Pseudomonas Vb3, Psychrobacter phenylpyruvicus, and Stenotrophomonas maltophilia were PYR negative. On the other hand Achromobacter piechaudii, Achromobacter denitrificans, Achromobacter xylosoxidans, Burkholderia gladioli, Chryseobacterium gleum-indologenes, Comamonas testosroni, Cupriavidus pauculus, Delftia acidovorans, Elizabethkingia meningoseptica, Myroides spp., Ochrobactrum anthropi, Pseudomonas oryzihabitans, Ralstonia pickettii, Rhizobium radiobacter, Shewanella spp., Sphingobacterium multivorum, Sphingobacterium spiritivorum, and Weeksella virosa were PYR positive. Finally, Acinetobacter lwoffii, Pseudomonas aeruginosa, Pseudomonas fluorescens, Roseomonas spp., and Sphingomonas paucimobilis-parapaucimobilis were PYR variable. PYR testing should be considered as a useful tool to facilitate the identification of NFGNR. PMID:16822636

  2. Effects of plant growth promoting rhizobacteria (PGPR) on rooting and root growth of kiwifruit (Actinidia deliciosa) stem cuttings.

    PubMed

    Erturk, Yasar; Ercisli, Sezai; Haznedar, Ayhan; Cakmakci, Ramazan

    2010-01-01

    The effects of plant growth promoting rhizobacteria (PGPR) on the rooting and root growth of semi-hardwood and hardwood kiwifruit stem cuttings were investigated. The PGPR used were Bacillus RC23, Paenibacillus polymyxa RC05, Bacillus subtilis OSU142, Bacillus RC03, Comamonas acidovorans RC41, Bacillus megaterium RC01 and Bacillus simplex RC19. All the bacteria showed indole-3-acetic acid (IAA) producing capacity. Among the PGPR used, the highest rooting ratios were obtained at 47.50% for semi-hardwood stem cuttings from Bacillus RC03 and Bacillus simplex RC19 treatments and 42.50% for hardwood stem cuttings from Bacillus RC03. As well, Comamonas acidovorans RC41 inoculations indicated higher value than control treatments. The results suggest that these PGPR can be used in organic nursery material production and point to the feasibility of synthetic auxin (IBA) replacement by organic management based on PGPR. PMID:21157636

  3. Effects of plant growth promoting rhizobacteria (PGPR) on rooting and root growth of kiwifruit (Actinidia deliciosa) stem cuttings.

    PubMed

    Erturk, Yasar; Ercisli, Sezai; Haznedar, Ayhan; Cakmakci, Ramazan

    2010-01-01

    The effects of plant growth promoting rhizobacteria (PGPR) on the rooting and root growth of semi-hardwood and hardwood kiwifruit stem cuttings were investigated. The PGPR used were Bacillus RC23, Paenibacillus polymyxa RC05, Bacillus subtilis OSU142, Bacillus RC03, Comamonas acidovorans RC41, Bacillus megaterium RC01 and Bacillus simplex RC19. All the bacteria showed indole-3-acetic acid (IAA) producing capacity. Among the PGPR used, the highest rooting ratios were obtained at 47.50% for semi-hardwood stem cuttings from Bacillus RC03 and Bacillus simplex RC19 treatments and 42.50% for hardwood stem cuttings from Bacillus RC03. As well, Comamonas acidovorans RC41 inoculations indicated higher value than control treatments. The results suggest that these PGPR can be used in organic nursery material production and point to the feasibility of synthetic auxin (IBA) replacement by organic management based on PGPR.

  4. Characterization of Microbial Communities in Gas Industry Pipelines

    PubMed Central

    Zhu, Xiang Y.; Lubeck, John; Kilbane, John J.

    2003-01-01

    Culture-independent techniques, denaturing gradient gel electrophoresis (DGGE) analysis, and random cloning of 16S rRNA gene sequences amplified from community DNA were used to determine the diversity of microbial communities in gas industry pipelines. Samples obtained from natural gas pipelines were used directly for DNA extraction, inoculated into sulfate-reducing bacterium medium, or used to inoculate a reactor that simulated a natural gas pipeline environment. The variable V2-V3 (average size, 384 bp) and V3-V6 (average size, 648 bp) regions of bacterial and archaeal 16S rRNA genes, respectively, were amplified from genomic DNA isolated from nine natural gas pipeline samples and analyzed. A total of 106 bacterial 16S rDNA sequences were derived from DGGE bands, and these formed three major clusters: beta and gamma subdivisions of Proteobacteria and gram-positive bacteria. The most frequently encountered bacterial species was Comamonas denitrificans, which was not previously reported to be associated with microbial communities found in gas pipelines or with microbially influenced corrosion. The 31 archaeal 16S rDNA sequences obtained in this study were all related to those of methanogens and phylogenetically fall into three clusters: order I, Methanobacteriales; order III, Methanomicrobiales; and order IV, Methanosarcinales. Further microbial ecology studies are needed to better understand the relationship among bacterial and archaeal groups and the involvement of these groups in the process of microbially influenced corrosion in order to develop improved ways of monitoring and controlling microbially influenced corrosion. PMID:12957923

  5. Bioaugmentation with a pyridine-degrading bacterium in a membrane bioreactor treating pharmaceutical wastewater.

    PubMed

    Wen, Donghui; Zhang, Jing; Xiong, Ruilin; Liu, Rui; Chen, Lujun

    2013-11-01

    The bacterial strain Paracoccus denitrificans W12, which could utilize pyridine as its sole source of carbon and nitrogen, was added into a membrane bioreactor (MBR) to enhance the treatment of a pharmaceutical wastewater. The treatment efficiencies investigated showed that the removal of chemical oxygen demand, total nitrogen, and total phosphorus were similar between bioaugmented and non-bioaugmented MBRs, however, significant removal of pyridine was obtained in the bioaugmented reactor. When the hydraulic retention time was 60 hr and the influent concentration of pyridine was 250-500 mg/L, the mean effluent concentration of pyridine without adding W12 was 57.2 mg/L, while the pyridine was degraded to an average of 10.2 mg/L with addition of W12. The bacterial community structure of activated sludge during the bioaugmented treatment was analyzed using polymerase chain reaction -denaturing gradient gel electrophoresis (PCR-DGGE). The results showed that the W12 inoculum reversed the decline of microbial community diversity, however, the similarity between bacterial community structure of the original sludge and that of the sludge after bioaugmentation decreased steadily during the wastewater treatment. Sequencing of the DNA recovered from DGGE gel indicated that Flavobacteriaceae sp., Sphingobium sp., Comamonas sp., and Hyphomicrobium sp. were the dominant organisms in time sequence in the bacterial community in the bioaugmented MBR. This implied that the bioaugmentation was affected by the adjustment of whole bacterial community structure in the inhospitable environment, rather than being due solely to the degradation performance of the bacterium added. PMID:24552055

  6. A Novel Testosterone Catabolic Pathway in Bacteria ▿ ‡

    PubMed Central

    Leu, Yann-Lii; Wang, Po-Hsiang; Shiao, Ming-Shi; Ismail, Wael; Chiang, Yin-Ru

    2011-01-01

    Forty years ago, Coulter and Talalay (A. W. Coulter and P. Talalay, J. Biol. Chem. 243:3238–3247, 1968) established the oxygenase-dependent pathway for the degradation of testosterone by aerobes. The oxic testosterone catabolic pathway involves several oxygen-dependent reactions and is not available for anaerobes. Since then, a variety of anaerobic bacteria have been described for the ability to degrade testosterone in the absence of oxygen. Here, a novel, oxygenase-independent testosterone catabolic pathway in such organisms is described. Steroidobacter denitrificansDSMZ18526 was shown to be capable of degrading testosterone in the absence of oxygen and was selected as the model organism in this study. In a previous investigation, we identified the initial intermediates involved in an anoxic testosterone catabolic pathway, most of which are identical to those of the oxic pathway demonstrated in Comamonas testosteroni. In this study, five additional intermediates of the anoxic pathway were identified. We demonstrated that subsequent steps of the anoxic pathway greatly differ from those of the established oxic pathway, which suggests that a novel pathway for testosterone catabolism is present. In the proposed anoxic pathway, a reduction reaction occurs at C-4 and C-5 of androsta-1,4-diene-3,17-dione, the last common intermediate of both the oxic and anoxic pathways. After that, a novel hydration reaction occurs and a hydroxyl group is thus introduced to the C-1α position of C19steroid substrates. To our knowledge, an enzymatic hydration reaction occurring at the A ring of steroid compounds has not been reported before. PMID:21725000

  7. ENVIROMENTALLY BENIGN MITIGATION OF MICROBIOLOGICALLY INFLUENCED CORROSION (MIC)

    SciTech Connect

    J. Robert Paterek; Gemma Husmillo; Amrutha Daram; Vesna Trbovic

    2003-10-31

    The overall program objective is to develop and evaluate environmentally benign agents or products that are effective in the prevention, inhibition, and mitigation of microbially influenced corrosion (MIC) in the internal surfaces of metallic natural gas pipelines. The goal is to develop one or more environmentally benign (a.k.a. ''green'') products that can be applied to maintain the structure and dependability of the natural gas infrastructure. The technical approach for this quarter includes the application of the method of fractionation of the extracts by high performance liquid chromatography (HPLC); determination of antimicrobial activities of the new extracts and fractions using a growth inhibition assay, and evaluation of the extracts' ability to inhibit biofilm formation. We initiated the delivery system for these new biocides in the test cell and in mixtures of foam components and biocides/anti-biofilms. A total of 51 fractions collected by HPLC from crude extracts that were obtained from three varieties of Capsicum sp. (Serrano, Habanero, Chile de Arbol) were subjected to growth inhibition tests against two SRB strains, D. vulgaris and D. desulfuricans. Five fractions showed growth inhibition against both strains while seven inhibited D. desulfuricans only. The crude extracts did not show growth inhibition on both strains but were proven to be potent in preventing the formation of biofilm. Growth inhibition tests of the same set of crude extracts against Comamonas denitrificans did not show positive results. The fractions will be subjected to biofilm inhibition and dissociation assay as well. The delivery system to be evaluated first was foam. The ''foam pig'' components of surfactants and water were tested with the biocide addition. The first chemical and physical parameters to be tested were pH and surfactants. Tests using the fractionated pepper extracts are progressing rapidly. Gas chromatographic analysis on a number of fractions is underway

  8. Bioaugmentation with a pyridine-degrading bacterium in a membrane bioreactor treating pharmaceutical wastewater.

    PubMed

    Wen, Donghui; Zhang, Jing; Xiong, Ruilin; Liu, Rui; Chen, Lujun

    2013-11-01

    The bacterial strain Paracoccus denitrificans W12, which could utilize pyridine as its sole source of carbon and nitrogen, was added into a membrane bioreactor (MBR) to enhance the treatment of a pharmaceutical wastewater. The treatment efficiencies investigated showed that the removal of chemical oxygen demand, total nitrogen, and total phosphorus were similar between bioaugmented and non-bioaugmented MBRs, however, significant removal of pyridine was obtained in the bioaugmented reactor. When the hydraulic retention time was 60 hr and the influent concentration of pyridine was 250-500 mg/L, the mean effluent concentration of pyridine without adding W12 was 57.2 mg/L, while the pyridine was degraded to an average of 10.2 mg/L with addition of W12. The bacterial community structure of activated sludge during the bioaugmented treatment was analyzed using polymerase chain reaction -denaturing gradient gel electrophoresis (PCR-DGGE). The results showed that the W12 inoculum reversed the decline of microbial community diversity, however, the similarity between bacterial community structure of the original sludge and that of the sludge after bioaugmentation decreased steadily during the wastewater treatment. Sequencing of the DNA recovered from DGGE gel indicated that Flavobacteriaceae sp., Sphingobium sp., Comamonas sp., and Hyphomicrobium sp. were the dominant organisms in time sequence in the bacterial community in the bioaugmented MBR. This implied that the bioaugmentation was affected by the adjustment of whole bacterial community structure in the inhospitable environment, rather than being due solely to the degradation performance of the bacterium added.

  9. Assessment of bacterial diversity during composting of agricultural byproducts

    PubMed Central

    2013-01-01

    Background Composting is microbial decomposition of biodegradable materials and it is governed by physicochemical, physiological and microbiological factors. The importance of microbial communities (bacteria, actinomycetes and fungi) during composting is well established. However, the microbial diversity during composting may vary with the variety of composting materials and nutrient supplements. Therefore, it is necessary to study the diversity of microorganisms during composting of different agricultural byproducts like wheat bran, rice bran, rice husk, along with grass clippings and bulking agents. Here it has been attempted to assess the diversity of culturable bacteria during composting of agricultural byproducts. Results The culturable bacterial diversity was assessed during the process by isolating the most prominent bacteria. Bacterial population was found to be maximum during the mesophilic phase, but decreased during the thermophilic phase and declined further in the cooling and maturation phase of composting. The bacterial population ranged from 105 to 109 cfu g-1 compost. The predominant bacteria were characterized biochemically, followed by 16S rRNA gene sequencing. The isolated strains, both Gram-positive and Gram-negative groups belonged to the order Burkholderiales, Enterobacteriales, Actinobacteriales and Bacillales, which includes genera e.g. Staphylococcus, Serratia, Klebsiella, Enterobacter, Terribacillus, Lysinibacillus Kocuria, Microbacterium, Acidovorax and Comamonas. Genera like Kocuria, Microbacterium, Acidovorax, Comamonas and some new species of Bacillus were also identified for the first time from the compost made from agricultural byproducts. Conclusion The use of appropriate nitrogen amendments and bulking agents in composting resulted in good quality compost. The culture based strategy enabled us to isolate some novel bacterial isolates like Kocuria, Microbacterium, Acidovorax and Comamonas first time from agro-byproducts compost

  10. Bacterial Diversity in Submarine Groundwater along the Coasts of the Yellow Sea.

    PubMed

    Ye, Qi; Liu, Jianan; Du, Jinzhou; Zhang, Jing

    2015-01-01

    Submarine groundwater (SGD) is one of the most significant pathways for the exchange of groundwater and/or source of nutrients, metals and carbon to the ocean, subsequently cause deleterious impacts on the coastal ecosystems. Microorganisms have been recognized as the important participators in the biogeochemical processes in the SGD. In this study, by utilizing 16S rRNA-based Illumina Miseq sequencing technology, we investigated bacterial diversity and distribution in both fresh well water and brackish recirculated porewater along the coasts in the Yellow Sea. The results showed that Actinobacteria and Betaproteobacteria, especially Comamonas spp. and Limnohabitans spp. were dominated in fresh well samples. Distinct patterns of bacterial communities were found among the porewater samples due to different locations, for examples, Cyanbacteria was the most abundant in the porewater samples far from the algal bloomed areas. The analysis of correlation between representative bacterial taxonomic groups and the contexture environmental parameters showed that fresh well water and brackish porewater might provide different nutrients to the coastal waters. Potential key bacterial groups such as Comamonas spp. may be excellent candidates for the bioremediation of the natural pollutants in the SGD. Our comprehensive understanding of bacterial diversity in the SGD along the coasts of the Yellow Sea will create a basis for designing the effective clean-up approach in-situ, and provide valuable information for the coastal management. PMID:26779172

  11. Bacterial Diversity in Submarine Groundwater along the Coasts of the Yellow Sea

    PubMed Central

    Ye, Qi; Liu, Jianan; Du, Jinzhou; Zhang, Jing

    2016-01-01

    Submarine groundwater (SGD) is one of the most significant pathways for the exchange of groundwater and/or source of nutrients, metals and carbon to the ocean, subsequently cause deleterious impacts on the coastal ecosystems. Microorganisms have been recognized as the important participators in the biogeochemical processes in the SGD. In this study, by utilizing 16S rRNA-based Illumina Miseq sequencing technology, we investigated bacterial diversity and distribution in both fresh well water and brackish recirculated porewater along the coasts in the Yellow Sea. The results showed that Actinobacteria and Betaproteobacteria, especially Comamonas spp. and Limnohabitans spp. were dominated in fresh well samples. Distinct patterns of bacterial communities were found among the porewater samples due to different locations, for examples, Cyanbacteria was the most abundant in the porewater samples far from the algal bloomed areas. The analysis of correlation between representative bacterial taxonomic groups and the contexture environmental parameters showed that fresh well water and brackish porewater might provide different nutrients to the coastal waters. Potential key bacterial groups such as Comamonas spp. may be excellent candidates for the bioremediation of the natural pollutants in the SGD. Our comprehensive understanding of bacterial diversity in the SGD along the coasts of the Yellow Sea will create a basis for designing the effective clean-up approach in-situ, and provide valuable information for the coastal management. PMID:26779172

  12. Bacterial Diversity in Submarine Groundwater along the Coasts of the Yellow Sea.

    PubMed

    Ye, Qi; Liu, Jianan; Du, Jinzhou; Zhang, Jing

    2015-01-01

    Submarine groundwater (SGD) is one of the most significant pathways for the exchange of groundwater and/or source of nutrients, metals and carbon to the ocean, subsequently cause deleterious impacts on the coastal ecosystems. Microorganisms have been recognized as the important participators in the biogeochemical processes in the SGD. In this study, by utilizing 16S rRNA-based Illumina Miseq sequencing technology, we investigated bacterial diversity and distribution in both fresh well water and brackish recirculated porewater along the coasts in the Yellow Sea. The results showed that Actinobacteria and Betaproteobacteria, especially Comamonas spp. and Limnohabitans spp. were dominated in fresh well samples. Distinct patterns of bacterial communities were found among the porewater samples due to different locations, for examples, Cyanbacteria was the most abundant in the porewater samples far from the algal bloomed areas. The analysis of correlation between representative bacterial taxonomic groups and the contexture environmental parameters showed that fresh well water and brackish porewater might provide different nutrients to the coastal waters. Potential key bacterial groups such as Comamonas spp. may be excellent candidates for the bioremediation of the natural pollutants in the SGD. Our comprehensive understanding of bacterial diversity in the SGD along the coasts of the Yellow Sea will create a basis for designing the effective clean-up approach in-situ, and provide valuable information for the coastal management.

  13. Foundation: Transforming data bases into knowledge bases

    NASA Technical Reports Server (NTRS)

    Purves, R. B.; Carnes, James R.; Cutts, Dannie E.

    1987-01-01

    One approach to transforming information stored in relational data bases into knowledge based representations and back again is described. This system, called Foundation, allows knowledge bases to take advantage of vast amounts of pre-existing data. A benefit of this approach is inspection, and even population, of data bases through an intelligent knowledge-based front-end.

  14. Michael hydratase alcohol dehydrogenase or just alcohol dehydrogenase?

    PubMed Central

    2014-01-01

    The Michael hydratase – alcohol dehydrogenase (MhyADH) from Alicycliphilus denitrificans was previously identified as a bi-functional enzyme performing a hydration of α,β-unsaturated ketones and subsequent oxidation of the formed alcohols. The investigations of the bi-functionality were based on a spectrophotometric assay and an activity staining in a native gel of the dehydrogenase. New insights in the recently discovered organocatalytic Michael addition of water led to the conclusion that the previously performed experiments to identify MhyADH as a bi-functional enzyme and their results need to be reconsidered and the reliability of the methodology used needs to be critically evaluated. PMID:24949265

  15. Microbial reduction of SO[sub 2] and NO[sub x] as a means of by- product recovery/disposal from regenerable processes for the desulfurization of flue gas

    SciTech Connect

    Sublette, K.L.

    1992-01-01

    Based on the work described simultaneous SO[sub 2]/No[sub x] removal from flue gas based on direct contact of the gas with SRB and T. denitrificans co-cultures or cultures-in-series has been eliminated as a viable process concept at this time. The technical reasons are as follows: (1) NO inhibition of SO[sub 2] reduction by D. desulfuricans - Although the NO concentrations used in the experiments described above are somewhat higher than that found in a typical flue gas, it is quite possible that at lower NO concentrations (or partial pressures) the inhibiting effects will simply take longer to become apparent. (2) Nitrate suppression of NO removal - As noted previously, the cultivation of T. denitrificans in a microbial flue gas treatment system (either one or two stages) would require sulfide-limiting conditions. Therefore, the electron acceptor must be in excess, requiring nitrate in the T. denitrificans process culture. As shown in experiments described above, nitrate significantly suppresses the removal of NO from a feed gas making simultaneous SO[sub 2]/NO[sub x] removal impractical by microbial means. (3) O[sub 2] inhibition of SO[sub 2] and NO reduction - It has been demonstrated that D. desulfuricans working cultures are tolerant of up to 1.7% O[sub 2] in the feed gas. However, further increases in the O[sub 2] partial pressure in the feed gas resulted in O[sub 2] inhibition of SO[sub 2] reduction. These inhibiting levels of O[sub 2] are comparable to those concentrations found in flue gases (3). Therefore, in any process in which raw flue gas contacts a D. desulfuricans culture marginal stability at best can be expected.

  16. Microbial reduction of SO{sub 2} and NO{sub x} as a means of by- product recovery/disposal from regenerable processes for the desulfurization of flue gas. Technical progress report, June 11, 1992--September 11, 1992

    SciTech Connect

    Sublette, K.L.

    1992-12-31

    Based on the work described simultaneous SO{sub 2}/No{sub x} removal from flue gas based on direct contact of the gas with SRB and T. denitrificans co-cultures or cultures-in-series has been eliminated as a viable process concept at this time. The technical reasons are as follows: (1) NO inhibition of SO{sub 2} reduction by D. desulfuricans - Although the NO concentrations used in the experiments described above are somewhat higher than that found in a typical flue gas, it is quite possible that at lower NO concentrations (or partial pressures) the inhibiting effects will simply take longer to become apparent. (2) Nitrate suppression of NO removal - As noted previously, the cultivation of T. denitrificans in a microbial flue gas treatment system (either one or two stages) would require sulfide-limiting conditions. Therefore, the electron acceptor must be in excess, requiring nitrate in the T. denitrificans process culture. As shown in experiments described above, nitrate significantly suppresses the removal of NO from a feed gas making simultaneous SO{sub 2}/NO{sub x} removal impractical by microbial means. (3) O{sub 2} inhibition of SO{sub 2} and NO reduction - It has been demonstrated that D. desulfuricans working cultures are tolerant of up to 1.7% O{sub 2} in the feed gas. However, further increases in the O{sub 2} partial pressure in the feed gas resulted in O{sub 2} inhibition of SO{sub 2} reduction. These inhibiting levels of O{sub 2} are comparable to those concentrations found in flue gases (3). Therefore, in any process in which raw flue gas contacts a D. desulfuricans culture marginal stability at best can be expected.

  17. Nickel-Resistant Bacteria from Anthropogenically Nickel-Polluted and Naturally Nickel-Percolated Ecosystems

    PubMed Central

    Stoppel, R.; Schlegel, H. G.

    1995-01-01

    DNA fragments harboring the nickel resistance determinants from bacteria isolated from anthropogenically polluted ecosystems in Europe and Zaire were compared with those harboring the nickel resistance determinants from bacteria isolated from naturally nickel-percolated soils from New Caledonia by DNA-DNA hybridization. The biotinylated DNA probes were derived from the previously described Alcaligenes eutrophus CH34, Alcaligenes xylosoxidans 31A, Alcaligenes denitrificans 4a-2, and Klebsiella oxytoca CCUG 15788 and four new nickel resistance-determining fragments cloned from strains isolated from soils under nickel-hyperaccumulating trees. Nine probes were hybridized with endonuclease-cleaved plasmid and total DNA samples from 56 nickel-resistant strains. Some of the New Caledonian strains were tentatively identified as Acinetobacter, Pseudomonas mendocina, Comamonas, Hafnia alvei, Burkholderia, Arthrobacter aurescens, and Arthrobacter ramosus strains. The DNA of most strains showed homologies to one or several of the following nickel resistance determinants: the cnr and ncc operons of the strains A. eutrophus CH34 and A. xylosoxidans 31A, respectively, the nre operon of strain 31A, and the nickel resistance determinants of K. oxytoca. On the basis of their hybridization reactions the nickel resistance determinants of the strains could be assigned to four groups: (i) cnr/ncc type, (ii) cnr/ncc/nre type, (iii) K. oxytoca type, and (iv) others. The majority of the strains were assigned to the known groups. Among the strains from Belgium and Zaire, exclusively the cnr/ncc and the cnr/ncc/nre types were found. Among the New Caledonian strains all four types were represented. Homologies to the nre operon were found only in combination with the cnr/ncc operon. The homologies to the cnr/ncc operon were the most abundant and were detected alone or together with homologies to the nre operon. Only the DNA of the strains isolated from soil in Scotland and the United States

  18. Structure and function of a hexameric copper-containing nitrite reductase.

    PubMed

    Nojiri, Masaki; Xie, Yong; Inoue, Tsuyoshi; Yamamoto, Takahiko; Matsumura, Hiroyoshi; Kataoka, Kunishige; Deligeer; Yamaguchi, Kazuya; Kai, Yasushi; Suzuki, Shinnichiro

    2007-03-13

    Dissimilatory nitrite reductase (NIR) is a key enzyme in denitrification, catalyzing the first step that leads to gaseous products (NO, N(2)O, and N(2)). We have determined the crystal structure of a Cu-containing NIR from a methylotrophic denitrifying bacterium, Hyphomicrobium denitrificans, at 2.2-A resolution. The overall structure of this H. denitrificans NIR reveals a trigonal prism-shaped molecule in which a monomer consisting of 447 residues and three Cu atoms is organized into a unique hexamer (i.e., a tightly associated dimer of trimers). Each monomer is composed of an N-terminal region containing a Greek key beta-barrel folding domain, cupredoxin domain I, and a C-terminal region containing cupredoxin domains II and III. Both cupredoxin domains I and II bind one type 1 Cu and are combined with a long loop comprising 31 amino acid residues. The type 2 Cu is ligated at the interface between domain II of one monomer and domain III of an adjacent monomer. Between the two trimeric C-terminal regions are three interfaces formed by an interaction between the domains I, and the type 1 Cu in the domain is required for dimerization of the trimer. The type 1 Cu in domain II functions as an electron acceptor from an electron donor protein and then transfers an electron to the type 2 Cu, binding the substrate to reduce nitrite to NO. The discussion of the intermolecular electron transfer process from cytochrome c(550) to the H. denitrificans NIR is based on x-ray crystallographic and kinetic results. PMID:17360521

  19. Water-based Screenprinting.

    ERIC Educational Resources Information Center

    Kreneck, Lynwood

    1989-01-01

    Outlines the techniques for silkscreening using water-based inks, concentrating on the qualities of water-based printing that differ from oil-based printing. Includes a step-by-step description of the process illustrated with photographs. (LS)

  20. Team-Based Learning

    ERIC Educational Resources Information Center

    Michaelsen, Larry K.; Sweet, Michael

    2011-01-01

    Team-based learning (TBL), when properly implemented, includes many, if not all, of the common elements of evidence-based best practices. To explain this, a brief overview of TBL is presented. The authors examine the relationship between the best practices of evidence-based teaching and the principles that constitute team-based learning. (Contains…

  1. Microbial Community Dynamics and Activity Link to Indigo Production from Indole in Bioaugmented Activated Sludge Systems

    PubMed Central

    Deng, Jie; Deng, Ye; Van Nostrand, Joy D.; Wu, Liyou; He, Zhili; Qin, Yujia; Zhou, Jiti; Zhou, Jizhong

    2015-01-01

    Biosynthesis of the popular dyestuff indigo from indole has been comprehensively studied using pure cultures, but less has been done to characterize the indigo production by microbial communities. In our previous studies, a wild strain Comamonas sp. MQ was isolated from activated sludge and the recombinant Escherichia coli nagAc carrying the naphthalene dioxygenase gene (nag) from strain MQ was constructed, both of which were capable of producing indigo from indole. Herein, three activated sludge systems, G1 (non-augmented control), G2 (augmented with Comamonas sp. MQ), and G3 (augmented with recombinant E. coli nagAc), were constructed to investigate indigo production. After 132-day operation, G3 produced the highest yields of indigo (99.5 ± 3.0 mg/l), followed by G2 (27.3 ± 1.3 mg/l) and G1 (19.2 ± 1.2 mg/l). The microbial community dynamics and activities associated with indigo production were analyzed by Illumina Miseq sequencing of 16S rRNA gene amplicons. The inoculated strain MQ survived for at least 30 days, whereas E. coli nagAc was undetectable shortly after inoculation. Quantitative real-time PCR analysis suggested the abundance of naphthalene dioxygenase gene (nagAc) from both inoculated strains was strongly correlated with indigo yields in early stages (0–30 days) (P < 0.001) but not in later stages (30–132 days) (P > 0.10) of operation. Based on detrended correspondence analysis (DCA) and dissimilarity test results, the communities underwent a noticeable shift during the operation. Among the four major genera (> 1% on average), the commonly reported indigo-producing populations Comamonas and Pseudomonas showed no positive relationship with indigo yields (P > 0.05) based on Pearson correlation test, while Alcaligenes and Aquamicrobium, rarely reported for indigo production, were positively correlated with indigo yields (P < 0.05). This study should provide new insights into our understanding of indigo bio-production by microbial communities

  2. Community-Based Care

    MedlinePlus

    ... our e-newsletter! Aging & Health A to Z Community-Based Care Basic Facts & Information A variety of ... Adult Day Care Adult day care is a community-based option that has become more common. It ...

  3. Carbon Based Nanotechnology: Review

    NASA Technical Reports Server (NTRS)

    Srivastava, Deepak; Saini, Subhash (Technical Monitor)

    1999-01-01

    This presentation reviews publicly available information related to carbon based nanotechnology. Topics covered include nanomechanics, carbon based electronics, nanodevice/materials applications, nanotube motors, nano-lithography and H2O storage in nanotubes.

  4. Stolen Base Physics

    NASA Astrophysics Data System (ADS)

    Kagan, David

    2013-05-01

    Few plays in baseball are as consistently close and exciting as the stolen base. While there are several studies of sprinting,2-4 the art of base stealing is much more nuanced. This article describes the motion of the base-stealing runner using a very basic kinematic model. The model will be compared to some data from a Major League game. The predictions of the model show consistency with the skills needed for effective base stealing.

  5. Stolen Base Physics

    ERIC Educational Resources Information Center

    Kagan, David

    2013-01-01

    Few plays in baseball are as consistently close and exciting as the stolen base. While there are several studies of sprinting, the art of base stealing is much more nuanced. This article describes the motion of the base-stealing runner using a very basic kinematic model. The model will be compared to some data from a Major League game. The…

  6. Volatile fatty acids influence on the structure of microbial communities producing PHAs

    PubMed Central

    Ciesielski, Slawomir; Przybylek, Grzegorz

    2014-01-01

    Polyhydroxyalkanoates (PHAs) can be produced by microorganisms and are a biodegradable alternative to fossil-fuel based plastics. Currently, the focus is on reducing production costs by exploring alternative substrates for PHAs production, and on producing copolymers which are less brittle than monomers. Accordingly, this study used a substrate consisting of wastewater from waste-glycerol fermentation, supplemented with different amounts of acetic and propionic acids. These substrates were used to feed mixed microbial communities enriched from activated sludge in a sequencing batch reactor. A reactor supplemented with 2 mL of acetic acid produced 227.8 mg/L of a homopolymer of hydroxybutyrate (3HB); 4 mL of acetic acid produced 279.8 mg/L 3HB; whereas 4 mL of propionic acid produced 673.0 mg/L of a copolymer of 3HB and 3HV (hydroxyvalerate). Ribosomal Intergenic Spacer Analysis (RISA) was used to show the differences between the communities created in the reactors. Thauera species predominated in biomass that produced 3HB; Paracoccus denitrificans in the biomass that produced 3HB-co-3HV. Because P. denitrificans produced the more desirable copolymer, it may be advantageous to promote its growth in PHAs-producing reactors by adding propionate. PMID:25242921

  7. Interdependence of two NarK domains in a fused nitrate/nitrite transporter.

    PubMed

    Goddard, Alan D; Moir, James W B; Richardson, David J; Ferguson, Stuart J

    2008-11-01

    Nitrate uptake is essential for various bacterial processes and combines with nitrite export to form the usual initial steps of denitrification, a process that reduces nitrate to dinitrogen gas. Although many bacterial species contain NarK-like transporters that are proposed to function as either nitrate/proton symporters or nitrate/nitrite antiporters based on sequence homology, these transporters remain, in general, poorly characterized. Several bacteria appear to contain a transporter that is a fusion of two NarK-like proteins, although the significance of this arrangement remains elusive. We demonstrate that NarK from Paracoccus denitrificans is expressed as a fusion of two NarK-like transporters. NarK1 and NarK2 are separately capable of supporting anaerobic denitrifying growth but with growth defects that are partially mitigated by coexpression of the two domains. NarK1 appears to be a nitrate/proton symporter with high affinity for nitrate and NarK2 a nitrate/nitrite antiporter with lower affinity for nitrate. Each transporter requires two conserved arginine residues for activity. A transporter consisting of inactivated NarK1 fused to active NarK2 has a dramatically increased affinity for nitrate compared with NarK2 alone, implying a functional interaction between the two domains. A potential model for nitrate and nitrite transport in P. denitrificans is proposed. PMID:18823285

  8. Performance of a full-scale modified anaerobic/anoxic/oxic process: High-throughput sequence analysis of its microbial structures and their community functions.

    PubMed

    Chen, Yasong; Zhao, Zheng; Peng, Yuke; Li, Jie; Xiao, Lin; Yang, Liuyan

    2016-11-01

    The average COD, TN, TP, and NH4(+)-N elimination rates in a new wastewater treatment plant (WWTP) based on a modified A2/O process were 83%, 72.4%, 93.5%, and 98.6%, respectively, even under conditions of a low C/N ratio and low temperature. Among the four potential denitrifying units, the post-anoxic unit was the least efficient with respect to the removal efficiency. However, the structures of the bacterial community among samples obtained from the treatment units were similar, as demonstrated using Illumina Miseq high-throughput sequencing. Genera with nitrifying, denitrifying, hydrolyzing, and glycogen-accumulating activities were identified in all units, indicating that functional groups were highly enriched in the active sludges and thus enabled nitrogen removal. The key functional microorganisms responsible for nitrification-denitrification in the WWTP were species belonging to the genera Nitrospira, Hydrogenophilaceae, Comamonas, Dechloromonas, Thauera, Haliangium, and Halomonas. PMID:27573476

  9. Saturn base heating handbook

    NASA Technical Reports Server (NTRS)

    Mullen, C. R.; Bender, R. L.; Bevill, R. L.; Reardon, J.; Hartley, L.

    1972-01-01

    A handbook containing a summary of model and flight test base heating data from the S-1, S-1B, S-4, S-1C, and S-2 stages is presented. A review of the available prediction methods is included. Experimental data are provided to make the handbook a single source of Saturn base heating data which can be used for preliminary base heating design predictions of launch vehicles.

  10. ARAC terrain data base

    SciTech Connect

    Walker, H.

    1982-11-01

    A terrain data base covering the continental United States at 500-meter resolution has been generated. Its function is to provide terrain data for input to mesoscale atmospheric models that are used as part of the Atmospheric Release Advisory Capability at Lawrence Livermore Laboratory (LLNL). The structure of the data base as it exists on the LLNL computer system is described. The data base has been written to tapes for transfer to other systems and the format of these tapes is also described.

  11. Action-Based Planning

    NASA Technical Reports Server (NTRS)

    Lansky, Amy L.; Lum, Henry Jr. (Technical Monitor)

    1994-01-01

    This paper presents an approach to domain representation and planning that is fundamentally different from traditional methods; an approach based strictly on actions and their interrelationships, rather than on state-based goals and preconditions. In particular, we focus on the action-based planner COLLAGE, describe its methods for plan-construction, and contrast them with more traditional approaches to planning. Experiences with COLLAGE in realistic domains have shown that the action-based approach is not only more natural to use, but can also be more cost-efficient than traditional planning methods.

  12. Understanding Acid Base Disorders.

    PubMed

    Gomez, Hernando; Kellum, John A

    2015-10-01

    The concentration of hydrogen ions is regulated in biologic solutions. There are currently 3 recognized approaches to assess changes in acid base status. First is the traditional Henderson-Hasselbalch approach, also called the physiologic approach, which uses the relationship between HCO3(-) and Pco2; the second is the standard base excess approach based on the Van Slyke equation. The third approach is the quantitative or Stewart approach, which uses the strong ion difference and the total weak acids. This article explores the origins of the current concepts framing the existing methods to analyze acid base balance.

  13. Synthetic Base Fluids

    NASA Astrophysics Data System (ADS)

    Brown, M.; Fotheringham, J. D.; Hoyes, T. J.; Mortier, R. M.; Orszulik, S. T.; Randles, S. J.; Stroud, P. M.

    The chemical nature and technology of the main synthetic lubricant base fluids is described, covering polyalphaolefins, alkylated aromatics, gas-to-liquid (GTL) base fluids, polybutenes, aliphatic diesters, polyolesters, polyalkylene glycols or PAGs and phosphate esters.Other synthetic lubricant base oils such as the silicones, borate esters, perfluoroethers and polyphenylene ethers are considered to have restricted applications due to either high cost or performance limitations and are not considered here.Each of the main synthetic base fluids is described for their chemical and physical properties, manufacture and production, their chemistry, key properties, applications and their implications when used in the environment.

  14. XML-BASED REPRESENTATION

    SciTech Connect

    R. KELSEY

    2001-02-01

    For focused applications with limited user and use application communities, XML can be the right choice for representation. It is easy to use, maintain, and extend and enjoys wide support in commercial and research sectors. When the knowledge and information to be represented is object-based and use of that knowledge and information is a high priority, then XML-based representation should be considered. This paper discusses some of the issues involved in using XML-based representation and presents an example application that successfully uses an XML-based representation.

  15. Base Oils from Petroleum

    NASA Astrophysics Data System (ADS)

    Prince, R. J.

    The source, composition and suitability of crude oils for base oil production are reviewed. The physical and chemical properties of alkanes, naphthenes and aromatics and their characteristics for lubricant applications are examined. Properties and applications of various base oils are defined and specified. Production of conventional mineral oils is described, including the various processes to remove wax and other deleterious substances, followed by increasingly severe hydrogenation to produce base oils of increased quality and performance. The API categorization of mineral base oils, either direct from the refinery or after hydrotreatment of increasing severity, is described, together with sub-categories.

  16. Soy-based polyols

    DOEpatents

    Suppes, Galen; Lozada, Zueica; Lubguban, Arnold

    2013-06-25

    The invention provides processes for preparing soy-based oligomeric polyols or substituted oligomeric polyols, as well as urethane bioelasteromers comprising the oligomeric polyols or substituted oligomeric polyols.

  17. Gasification-based biomass

    SciTech Connect

    None, None

    2009-01-18

    The gasification-based biomass section of the Renewable Energy Technology Characterizations describes the technical and economic status of this emerging renewable energy option for electricity supply.

  18. Diagnosis and quantification of glycerol assimilating denitrifying bacteria in an integrated fixed-film activated sludge reactor via 13C DNA stable-isotope probing.

    PubMed

    Lu, Huijie; Chandran, Kartik

    2010-12-01

    Glycerol, a byproduct of biodiesel and oleo-chemical manufacturing operations, represents an attractive alternate to methanol as a carbon and electron donor for enhanced denitrification. However, unlike methanol, little is known about the diversity and activity of glycerol assimilating bacteria in activated sludge. In this study, the microbial ecology of glycerol assimilating denitrifying bacteria in a sequencing batch integrated fixed film activated sludge (SB-IFAS) reactor was investigated using (13)C-DNA stable isotope probing (SIP). During steady state SB-IFAS reactor operation, near complete nitrate removal (92.7 ± 5.8%) was achieved. Based on (13)C DNA clone libraries obtained after 360 days of SB-IFAS reactor operation, bacteria related to Comamonas spp. and Diaphorobacter spp. dominated in the suspended phase communities. (13)C assimilating members in the biofilm community were phylogenetically more diverse and were related to Comamonas spp., Bradyrhizobium spp., and Tessaracoccus spp. Possibly owing to greater substrate availability in the suspended phase, the glycerol-assimilating denitrifying populations (quantified by real-time PCR) were more abundant therein than in the biofilm phase. The biomass in the suspended phase also had a higher specific denitrification rate than the biofilm phase (p = 4.33e-4), and contributed to 69.7 ± 4.5% of the overall N-removal on a mass basis. The kinetics of glycerol based denitrification by suspended phase biomass were approximately 3 times higher than with methanol. Previously identified methanol assimilating denitrifying bacteria were not associated with glycerol assimilation, thereby suggesting limited cross-utilization of these two substrates for denitrification in the system tested.

  19. Scientifically Based Research.

    ERIC Educational Resources Information Center

    Beghetto, Ron

    2003-01-01

    Most principals are aware that the No Child Left Behind Act of 2001 makes it mandatory for school leaders who depend on federal funding to select and implement programs that are based on scientific research. This publication reviews five documents that offer insights into what is meant by scientifically based research and help school leaders…

  20. Content-Based Instruction.

    ERIC Educational Resources Information Center

    CATESOL Journal, 1992

    1992-01-01

    This English-as-a-Second Language (ESL) journal periodical devotes entire issues to specific issues. The theme of this issue is "Content-Base Instruction." Articles include: "Syllabus Design in Content-Based Instruction" (David E. Eskey); "How Relevant Is Relevance?: An Examination of Student Needs, Interests, and Motivation in the Content-Based…

  1. Medical Knowledge Bases.

    ERIC Educational Resources Information Center

    Miller, Randolph A.; Giuse, Nunzia B.

    1991-01-01

    Few commonly available, successful computer-based tools exist in medical informatics. Faculty expertise can be included in computer-based medical information systems. Computers allow dynamic recombination of knowledge to answer questions unanswerable with print textbooks. Such systems can also create stronger ties between academic and clinical…

  2. Management Data Base Development.

    ERIC Educational Resources Information Center

    Dan, Robert L.

    1975-01-01

    A management data base is seen as essential for a management information system, program budgeting, program costing, management by objectives, program evaluation, productivity measures, and accountability in institutions of higher education. The necessity of a management data base is addressed, along with the benefits and limitations it may have…

  3. Propagation data bases

    NASA Astrophysics Data System (ADS)

    1981-12-01

    Existing data bases accumulated as the result of experiments to gather propagation data on millimeter wave Earth-space links are described. The satellites used are described and results of the significant experiments conducted in the United States are summarized. The data bases consist primarily of cumulative attenuation statistics, though some depolarization measurements are included. Additional summaries of propagation data are cited.

  4. Content-Based Instruction

    ERIC Educational Resources Information Center

    DelliCarpini, M.; Alonso, O.

    2013-01-01

    DelliCarpini and Alonso's book "Content-Based Instruction" explores different approaches to teaching content-based instruction (CBI) in the English language classroom. They provide a comprehensive overview of how to teach CBI in an easy-to-follow guide that language teachers will find very practical for their own contexts. Topics…

  5. Red-based cumulus.

    PubMed

    Gedzelman, Stanley David

    2015-02-01

    Observations and model simulations of cumulus clouds whose bases are tinted red when the Sun is well above the horizon are presented. Conditions for seeing red bases include (1) a red underlying surface (which may consist of dust clouds, as from haboobs) with high albedo, (2) small fractional cloud cover when the Sun is far enough below the zenith for direct sunlight to illuminate much of the surface directly below and around cloud base, (3) optically thick clouds so that the bases are dark, and (4) clouds with bases that are near enough to the observer to appear high in the sky so that the admixture of scattered light from the intervening atmosphere is minimized.

  6. Treatment of cosmetic effluent in different configurations of ceramic UF membrane based bioreactor: Toxicity evaluation of the untreated and treated wastewater using catfish (Heteropneustes fossilis).

    PubMed

    Banerjee, Priya; Dey, Tanmoy Kumar; Sarkar, Sandeep; Swarnakar, Snehasikta; Mukhopadhyay, Aniruddha; Ghosh, Sourja

    2016-03-01

    Extensive usage of pharmaceutical and personal care products (PPCPs) and their discharge through domestic sewage have been recently recognized as a new generation environmental concern which deserves more scientific attention over the classical environmental pollutants. The major issues of this type of effluent addressed in this study were its colour, triclosan and anionic surfactant (SDS) content. Samples of cosmetic effluent were collected from different beauty treatment salons and spas in and around Kolkata, India and treated in bioreactors containing a bacterial consortium isolated from activated sludge samples collected from a common effluent treatment plant. Members of the consortium were isolated and identified as Klebsiella sp., Pseudomonas sp., Salmonella sp. and Comamonas sp. The biotreated effluent was subjected to ultrafiltration (UF) involving indigenously prepared ceramic membranes in both side-stream and submerged mode. Analysis of the MBR treated effluent revealed 99.22%, 98.56% and 99.74% removal of colour, triclosan and surfactant respectively. Investigation of probable acute and chronic cyto-genotoxic potential of the untreated and treated effluents along with their possible participation in triggering oxidative stress was carried out with Heteropneustes fossilis (Bloch). Comet formation recorded in both liver and gill cells and micronucleus count in peripheral erythrocytes of individuals exposed to untreated effluent increased with duration of exposure and was significantly higher than those treated with UF permeates which in turn neared control levels. Results of this study revealed successful application of the isolated bacterial consortium in MBR process for efficient detoxification of cosmetic effluent thereby conferring the same suitable for discharge and/or reuse. PMID:26714296

  7. Skull Base Anatomy.

    PubMed

    Patel, Chirag R; Fernandez-Miranda, Juan C; Wang, Wei-Hsin; Wang, Eric W

    2016-02-01

    The anatomy of the skull base is complex with multiple neurovascular structures in a small space. Understanding all of the intricate relationships begins with understanding the anatomy of the sphenoid bone. The cavernous sinus contains the carotid artery and some of its branches; cranial nerves III, IV, VI, and V1; and transmits venous blood from multiple sources. The anterior skull base extends to the frontal sinus and is important to understand for sinus surgery and sinonasal malignancies. The clivus protects the brainstem and posterior cranial fossa. A thorough appreciation of the anatomy of these various areas allows for endoscopic endonasal approaches to the skull base.

  8. Data base management study

    NASA Technical Reports Server (NTRS)

    1976-01-01

    Data base management techniques and applicable equipment are described. Recommendations which will assist potential NASA data users in selecting and using appropriate data base management tools and techniques are presented. Classes of currently available data processing equipment ranging from basic terminals to large minicomputer systems were surveyed as they apply to the needs of potential SEASAT data users. Cost and capabilities projections for this equipment through 1985 were presented. A test of a typical data base management system was described, as well as the results of this test and recommendations to assist potential users in determining when such a system is appropriate for their needs. The representative system tested was UNIVAC's DMS 1100.

  9. Cyclodextrin-based molecular machines.

    PubMed

    Hashidzume, Akihito; Yamaguchi, Hiroyasu; Harada, Akira

    2014-01-01

    This chapter overviews molecular machines based on cyclodextrins (CDs). The categories of CD-based molecular machines, external stimuli for CD-based molecular machines, and typical examples of CD-based molecular machines are briefly described.

  10. Illumina MiSeq Sequencing Reveals Diverse Microbial Communities of Activated Sludge Systems Stimulated by Different Aromatics for Indigo Biosynthesis from Indole.

    PubMed

    Zhang, Xuwang; Qu, Yuanyuan; Ma, Qiao; Zhang, Zhaojing; Li, Duanxing; Wang, Jingwei; Shen, Wenli; Shen, E; Zhou, Jiti

    2015-01-01

    Indole, as a typical N-heteroaromatic compound existed in coking wastewater, can be used for bio-indigo production. The microbial production of indigo from indole has been widely reported during the last decades using culture-dependent methods, but few studies have been carried out by microbial communities. Herein, three activated sludge systems stimulated by different aromatics, i.e. naphthalene plus indole (G1), phenol plus indole (G2) and indole only (G3), were constructed for indigo production from indole. During the operation, G1 produced the highest indigo yield in the early stage, but it switched to G3 in the late stage. Based on LC-MS analysis, indigo was the major product in G1 and G3, while the purple product 2-(7-oxo-1H-indol-6(7H)-ylidene) indolin-3-one was dominant in G2. Illumina MiSeq sequencing of 16S rRNA gene amplicons was applied to analyze the microbial community structure and composition. Detrended correspondence analysis (DCA) and dissimilarity tests showed that the overall community structures of three groups changed significantly during the operation (P<0.05). Nevertheless, the bacteria assigned to phylum Proteobacteria, family Comamonadaceae, and genera Diaphorobacter, Comamonas and Aquamicrobium were commonly shared dominant populations. Pearson correlations were calculated to discern the relationship between microbial communities and indigo yields. The typical indigo-producing populations Comamonas and Pseudomonas showed no positive correlations with indigo yields, while there emerged many other genera that exhibited positive relationships, such as Aquamicrobium, Truepera and Pusillimonas, which had not been reported for indigo production previously. The present study should provide new insights into indigo bio-production by microbial communities from indole.

  11. Illumina MiSeq Sequencing Reveals Diverse Microbial Communities of Activated Sludge Systems Stimulated by Different Aromatics for Indigo Biosynthesis from Indole

    PubMed Central

    Zhang, Xuwang; Qu, Yuanyuan; Ma, Qiao; Zhang, Zhaojing; Li, Duanxing; Wang, Jingwei; Shen, Wenli; Shen, E; Zhou, Jiti

    2015-01-01

    Indole, as a typical N-heteroaromatic compound existed in coking wastewater, can be used for bio-indigo production. The microbial production of indigo from indole has been widely reported during the last decades using culture-dependent methods, but few studies have been carried out by microbial communities. Herein, three activated sludge systems stimulated by different aromatics, i.e. naphthalene plus indole (G1), phenol plus indole (G2) and indole only (G3), were constructed for indigo production from indole. During the operation, G1 produced the highest indigo yield in the early stage, but it switched to G3 in the late stage. Based on LC-MS analysis, indigo was the major product in G1 and G3, while the purple product 2-(7-oxo-1H-indol-6(7H)-ylidene) indolin-3-one was dominant in G2. Illumina MiSeq sequencing of 16S rRNA gene amplicons was applied to analyze the microbial community structure and composition. Detrended correspondence analysis (DCA) and dissimilarity tests showed that the overall community structures of three groups changed significantly during the operation (P<0.05). Nevertheless, the bacteria assigned to phylum Proteobacteria, family Comamonadaceae, and genera Diaphorobacter, Comamonas and Aquamicrobium were commonly shared dominant populations. Pearson correlations were calculated to discern the relationship between microbial communities and indigo yields. The typical indigo-producing populations Comamonas and Pseudomonas showed no positive correlations with indigo yields, while there emerged many other genera that exhibited positive relationships, such as Aquamicrobium, Truepera and Pusillimonas, which had not been reported for indigo production previously. The present study should provide new insights into indigo bio-production by microbial communities from indole. PMID:25928424

  12. Event-Based Science.

    ERIC Educational Resources Information Center

    Wright, Russell G.

    1992-01-01

    Suggests that an event-based science curriculum can provide the framework for deciding what to retain in an overloaded science curriculum. Provides examples of current events and the science concepts explored related to the event. (MDH)

  13. Navigating Value Based Care.

    PubMed

    Sorrel, Amy Lynn

    2015-12-01

    TMA is collaborating with TMF Health Quality Institute to connect Texas physicians to free TMF resources that will better position doctors for the rapid transition to value-based payment. PMID:26630238

  14. Benzimidazole Based Aerogel Materials

    NASA Technical Reports Server (NTRS)

    Rhine, Wendell E. (Inventor); Mihalcik, David (Inventor)

    2016-01-01

    The present invention provides aerogel materials based on imidazoles and polyimidazoles. The polyimidazole based aerogel materials can be thermally stable up to 500 C or more, and can be carbonized to produce a carbon aerogel having a char yield of 60% or more, specifically 70% or more. The present invention also provides methods of producing polyimidazole based aerogel materials by reacting at least one monomer in a suitable solvent to form a polybenzimidazole gel precursor solution, casting the polybenzimidazole gel precursor solution into a fiber reinforcement phase, allowing the at least one gel precursor in the precursor solution to transition into a gel material, and drying the gel materials to remove at least a portion of the solvent, to obtain an polybenzimidazole-based aerogel material.

  15. Meditation-Based Training

    PubMed Central

    2008-01-01

    Recent studies suggest that training-based measures are effective in improving cognitive skills. Meditation-based training has produced lasting changes in brain and cognitive functions. This technique of mental training exhibits neuroplasticity in the attentional networks, exhibiting superior performance, especially in the domain of attention and executive control processing, which is impaired in attention deficit hyperactivity disorder (ADHD). Although intervention techniques for ADHD are well researched, many individuals continue to experience significant functional impairment despite the symptom improvement. This emphasizes a need for a comprehensive approach that requires an effective behavioral intervention. The present paper provides a converging review of meditation-based effects on the brain, dysfunctions of ADHD, and suggestions for enhancement of cognitive abilities in patients with ADHD using meditational training combined with existing measures of intervention. The idea proposed herein should be considered a step for initiation of empirical studies on meditation-based training intervention and outcome effects. PMID:19727310

  16. Transformation based endorsement systems

    NASA Technical Reports Server (NTRS)

    Sudkamp, Thomas

    1988-01-01

    Evidential reasoning techniques classically represent support for a hypothesis by a numeric value or an evidential interval. The combination of support is performed by an arithmetic rule which often requires restrictions to be placed on the set of possibilities. These assumptions usually require the hypotheses to be exhausitive and mutually exclusive. Endorsement based classification systems represent support for the alternatives symbolically rather than numerically. A framework for constructing endorsement systems is presented in which transformations are defined to generate and update the knowledge base. The interaction of the knowledge base and transformations produces a non-monotonic reasoning system. Two endorsement based reasoning systems are presented to demonstrate the flexibility of the transformational approach for reasoning with ambiguous and inconsistent information.

  17. COMMUNITY BASED ENVIRONMENTAL PROTECTION

    EPA Science Inventory

    Community Based Environmental Protection intends to make environmental protection spring from the needs and values of the community of interest. Real community involvement in protecting the environment requires a process in which the environmental needs of communities and ecosyst...

  18. SLM based semiconductor maskwriter

    NASA Astrophysics Data System (ADS)

    Diez, Steffen; Jehle, Achim

    2015-09-01

    The high-end semiconductor mask fabrication is dominated by e-beam technology. But still more than 50% of all semiconductor masks are produced by laser writers. The current laser writers are based on the same technology that was used 25 years ago. They are reliable and fast but not very economical. Heidelberg Instruments has developed a new economical and fast laser writer based on the latest technologies.

  19. New formaldehyde base disinfectants.

    NASA Technical Reports Server (NTRS)

    Trujillo, R.; Lindell, K. F.

    1973-01-01

    Preparations of formaldehyde in various organic liquids - ethylene glycol, glycerol, and propylene glycol - serve as effective disinfectants towards microbial vegetative cells and spores. This disinfection is a temperature-dependent process and is manifest when these formaldehyde base disinfectants are dissolved in water. The irritating vapors associated with formaldehyde disinfection are not present in either of these new formaldehyde base disinfectants or in aqueous solutions of them.

  20. LDEF materials data bases

    NASA Technical Reports Server (NTRS)

    Funk, Joan G.; Strickland, John W.; Davis, John M.

    1993-01-01

    The Long Duration Exposure Facility (LDEF) and the accompanying experiments were composed of and contained a wide variety of materials representing the largest collection of materials flown in low Earth orbit (LEO) and retrieved for ground based analysis to date. The results and implications of the mechanical, thermal, optical, and electrical data from these materials are the foundation on which future LEO space missions will be built. The LDEF Materials Special Investigation Group (MSIG) has been charged with establishing and developing data bases to document these materials and their performance to assure not only that the data are archived for future generations but also that the data are available to the spacecraft user community in an easily accessed, user-friendly form. This paper discusses the format and content of the three data bases developed or being developed to accomplish this task. The hardware and software requirements for each of these three data bases are discussed along with current availability of the data bases. This paper also serves as a user's guide to the MAPTIS LDEF Materials Data Base.

  1. Swarm-Based Medicine

    PubMed Central

    Oldenburg, Jan

    2013-01-01

    Occasionally, medical decisions have to be taken in the absence of evidence-based guidelines. Other sources can be drawn upon to fill in the gaps, including experience and intuition. Authorities or experts, with their knowledge and experience, may provide further input—known as “eminence-based medicine”. Due to the Internet and digital media, interactions among physicians now take place at a higher rate than ever before. With the rising number of interconnected individuals and their communication capabilities, the medical community is obtaining the properties of a swarm. The way individual physicians act depends on other physicians; medical societies act based on their members. Swarm behavior might facilitate the generation and distribution of knowledge as an unconscious process. As such, “swarm-based medicine” may add a further source of information to the classical approaches of evidence- and eminence-based medicine. How to integrate swarm-based medicine into practice is left to the individual physician, but even this decision will be influenced by the swarm. PMID:24052454

  2. Plant based butters.

    PubMed

    Gorrepati, Kalyani; Balasubramanian, S; Chandra, Pitam

    2015-07-01

    During the last few years the popularity for the plant based butters (nut and seed butters) has increased considerably. Earlier peanut butter was the only alternative to the dairy butter, but over the years development in the technologies and also the consumer awareness about the plant based butters, has led the development of myriad varieties of butters with different nuts and seeds, which are very good source of protein, fiber, essential fatty acids and other nutrients. These days' different varieties of plant based butters are available in the market viz., peanut butter, soy butter, almond butter, pistachio butter, cashew butter and sesame butter etc. The form of butter is one of the healthy way of integrating nuts and seeds in to our regular diet. Nut and seed butters are generally prepared by roasting, grinding and refrigerated to consume it when it is still fresh. During this process it is imperative to retain the nutritional properties of these nuts and seeds in order to reap the benefits of the fresh nuts and seeds in the form of butter as well. Proper care is needed to minimize the conversion of healthful components in to unhealthy components during processing and further storage. Roasting temperature, temperatures during grinding and storage are the vital factors to be considered in order to have healthy and nutritious plant based butters. In this article, different plant based butters and their processing methods have been described. PMID:26139864

  3. Anterior skull base oncocytoma.

    PubMed

    López, Fernando; Vivanco, Blanca; Suárez, Carlos; Llorente, José L

    2013-03-01

    Oncocytic neoplasms are tumors composed of oncocytes (ie, epithelial cells with a large cytoplasm that is rich in mitochondria). Most cases are benign and originate from the salivary glands. Although there have been a few reported cases of oncocytomas being found in the sinonasal tract, most if not all cases seem not to involve the anterior skull base. We report a rare case of oncocytoma involving the anterior skull base occurring in a 44-year-old male patient. Preoperative carotid angiography and selective embolization was performed. The patient underwent an expanded endoscopic endonasal anterior craniofacial resection, which allowed complete resection of the tumor, with a low morbidity. The pathological diagnosis was oncocytoma. At 36 months after the initial treatment, the patient is free of disease. Based on our literature search, this may be the first such reported case. A brief review of the available literature examining the known body of knowledge regarding these neoplasms is presented.

  4. Lidar base specification

    USGS Publications Warehouse

    Heidemann, Hans Karl.

    2012-01-01

    Lidar is a fast evolving technology, and much has changed in the industry since the final draft of the “Lidar Base Specification Version 1.0” was written. Lidar data have improved in accuracy and spatial resolution, geospatial accuracy standards have been revised by the American Society for Photogrammetry and Remote Sensing (ASPRS), industry standard file formats have been expanded, additional applications for lidar have become accepted, and the need for interoperable data across collections has been realized. This revision to the “Lidar Base Specification Version 1.0” publication addresses those changes and provides continued guidance towards a nationally consistent lidar dataset.

  5. Protein based Block Copolymers

    PubMed Central

    Rabotyagova, Olena S.; Cebe, Peggy; Kaplan, David L.

    2011-01-01

    Advances in genetic engineering have led to the synthesis of protein-based block copolymers with control of chemistry and molecular weight, resulting in unique physical and biological properties. The benefits from incorporating peptide blocks into copolymer designs arise from the fundamental properties of proteins to adopt ordered conformations and to undergo self-assembly, providing control over structure formation at various length scales when compared to conventional block copolymers. This review covers the synthesis, structure, assembly, properties, and applications of protein-based block copolymers. PMID:21235251

  6. Process-based costing.

    PubMed

    Lee, Robert H; Bott, Marjorie J; Forbes, Sarah; Redford, Linda; Swagerty, Daniel L; Taunton, Roma Lee

    2003-01-01

    Understanding how quality improvement affects costs is important. Unfortunately, low-cost, reliable ways of measuring direct costs are scarce. This article builds on the principles of process improvement to develop a costing strategy that meets both criteria. Process-based costing has 4 steps: developing a flowchart, estimating resource use, valuing resources, and calculating direct costs. To illustrate the technique, this article uses it to cost the care planning process in 3 long-term care facilities. We conclude that process-based costing is easy to implement; generates reliable, valid data; and allows nursing managers to assess the costs of new or modified processes.

  7. Model based manipulator control

    NASA Technical Reports Server (NTRS)

    Petrosky, Lyman J.; Oppenheim, Irving J.

    1989-01-01

    The feasibility of using model based control (MBC) for robotic manipulators was investigated. A double inverted pendulum system was constructed as the experimental system for a general study of dynamically stable manipulation. The original interest in dynamically stable systems was driven by the objective of high vertical reach (balancing), and the planning of inertially favorable trajectories for force and payload demands. The model-based control approach is described and the results of experimental tests are summarized. Results directly demonstrate that MBC can provide stable control at all speeds of operation and support operations requiring dynamic stability such as balancing. The application of MBC to systems with flexible links is also discussed.

  8. Performance-Based Assessment

    ERIC Educational Resources Information Center

    ERIC Review, 1994

    1994-01-01

    "The ERIC Review" is published three times a year and announces research results, publications, and new programs relevant to each issue's theme topic. This issue explores performance-based assessment via two principal articles: "Performance Assessment" (Lawrence M. Rudner and Carol Boston); and "Alternative Assessment: Implications for Social…

  9. Protein Crystal Based Nanomaterials

    NASA Technical Reports Server (NTRS)

    Bell, Jeffrey A.; VanRoey, Patrick

    2001-01-01

    This is the final report on a NASA Grant. It concerns a description of work done, which includes: (1) Protein crystals cross-linked to form fibers; (2) Engineering of protein to favor crystallization; (3) Better knowledge-based potentials for protein-protein contacts; (4) Simulation of protein crystallization.

  10. Space based OTV servicing

    NASA Technical Reports Server (NTRS)

    Mcallister, J. G.

    1984-01-01

    Space based servicing of an orbit transfer vehicle (OTV) was previously outlined in sufficient detail to arrive at OTV and support system servicing requirements. Needed space station facilities and their functional requirements were identified. The impact of logistics and space serviceable design on the OTV design is detailed herein. RL10 derivative rocket engine inspection task times are enumerated.

  11. Developing Mobile Based Instruction

    ERIC Educational Resources Information Center

    Martin, Florence; Pastore, Raymond; Snider, Jean

    2012-01-01

    This paper describes an instructional design class's experience developing instruction for the mobile web. The class was taught at a southeastern university in the United States in a master's level computer based instruction course. Two example projects are showcased and student reflections on design issues are highlighted. Additionally,…

  12. Landscape-based Indicators

    EPA Science Inventory

    The report is based on data and experience gained through the GLNPO-funded Great Lakes Coastal Wetland Consortium (GLCWC) and the EPA-STAR funded Great Lakes Ecological Indicators Project (GLEI). EPA-MED author Trebitz and other MED personnel were collaborators on the GLEI proje...

  13. Drafting. Competency Based Curriculum.

    ERIC Educational Resources Information Center

    Everly, Al; And Others

    This competency based drafting curriculum is presented in seven specialization sections with units in each section containing a competency statement, performance objective, learning activities, evaluation, and quiz or problem sheets. Some units also contain answer sheets and/or handout sheets. Sections and number of units presented are (1) basic…

  14. Resource-Based Learning.

    ERIC Educational Resources Information Center

    Brown, Sally, Ed.; Smith, Brenda, Ed.

    The selections in this book encompass a broad spectrum of resource-based learning experiences, and are intended to help teachers and administrators gain a better understanding of the concepts and devise effective and efficient ways to use these materials. Titles include: "Introducing Resources for Learning" (Sally Brown and Brenda Smith);…

  15. Lunar Base Sitting

    NASA Technical Reports Server (NTRS)

    Staehle, Robert L.; Burke, James D.; Snyder, Gerald C.; Dowling, Richard; Spudis, Paul D.

    1993-01-01

    Speculation with regard to a permanent lunar base has been with us since Robert Goddard was working on the first liquid-fueled rockets in the 1920's. With the infusion of data from the Apollo Moon flights, a once speculative area of space exploration has become an exciting possibility. A Moon base is not only a very real possibility, but is probably a critical element in the continuation of our piloted space program. This article, originally drafted by World Space Foundation volunteers in conjuction with various academic and research groups, examines some of the strategies involved in selecting an appropriate site for such a lunar base. Site selection involves a number of complex variables, including raw materials for possible rocket propellant generation, hot an cold cycles, view of the sky (for astronomical considerations, among others), geological makeup of the region, and more. This article summarizes the key base siting considerations and suggests some alternatives. Availability of specific resources, including energy and certain minerals, is critical to success.

  16. "Comunidades de Base" Schools.

    ERIC Educational Resources Information Center

    Ozar, Lorraine A.

    1993-01-01

    Describes the objectives of a proposal to create two Catholic "base community" schools in Hispanic communities in Chicago, which will allow parents full participation and infuse neighborhoods with hope. Discusses several broad assumptions about the role of the schools and steps to ensure schools are products of the community. (BCY)

  17. Team Based Work. Symposium.

    ERIC Educational Resources Information Center

    2002

    This document contains three papers from a symposium on team-based work in human resource development (HRD). "Toward Transformational Learning in Organizations: Effects of Model-II Governing Variables on Perceived Learning in Teams" (Blair K. Carruth) summarizes a study that indicated that, regardless of which Model-II variable (valid information,…

  18. Service-based Solutions.

    ERIC Educational Resources Information Center

    Cummings, Lynda; Winston, Michael

    1998-01-01

    Describes the Solutions model used at Shelley High School in Idaho which gives students the opportunity to gain practical experience while tackling community problems. This approach is built on the three fundamentals of an integrated curriculum, a problem-solving focus, and service-based learning. Sample problems include increasing certain trout…

  19. SUPERCONDUCTING VANADIUM BASE ALLOY

    DOEpatents

    Cleary, H.J.

    1958-10-21

    A new vanadium-base alloy which possesses remarkable superconducting properties is presented. The alloy consists of approximately one atomic percent of palladium, the balance being vanadium. The alloy is stated to be useful in a cryotron in digital computer circuits.

  20. Project-Based Science

    ERIC Educational Resources Information Center

    Krajcik, Joe

    2015-01-01

    Project-based science is an exciting way to teach science that aligns with the "Next Generation Science Standards" ("NGSS"). By focusing on core ideas along with practices and crosscutting concepts, classrooms become learning environments where teachers and students engage in science by designing and carrying out…

  1. Concept-Based Learning

    ERIC Educational Resources Information Center

    Schill, Bethany; Howell, Linda

    2011-01-01

    A major part of developing concept-based instruction is the use of an overarching idea to provide a conceptual lens through which students view the content of a particular subject. By using a conceptual lens to focus learning, students think at a much deeper level about the content and its facts (Erickson 2007). Therefore, the authors collaborated…

  2. WATERSHED BASED SURVEY DESIGNS

    EPA Science Inventory

    The development of watershed-based design and assessment tools will help to serve the multiple goals for water quality monitoring required under the Clean Water Act, including assessment of regional condition to meet Section 305(b), identification of impaired water bodies or wate...

  3. Resource-Based Learning.

    ERIC Educational Resources Information Center

    Thomas, Margie Klink

    1999-01-01

    Provides an annotated bibliography of publications related to resource-based learning, which is defined as a student-centered learning environment grounded in learning theory in which the teacher and the library-media specialist collaborate to help students with information needs, information retrieval, analyzing and synthesizing the information,…

  4. Computer Based Library Orientation.

    ERIC Educational Resources Information Center

    Machalow, Robert

    This document presents computer-based lessons used to teach basic library skills to college students at York College of the City University of New York. The information for library orientation has been entered on a disk which must be used in conjunction with a word processing program, the Applewriter IIe, and an Apple IIe microcomputer. The…

  5. A Mars base

    NASA Technical Reports Server (NTRS)

    Soule, Veronique

    1989-01-01

    This study was initiated to provide an approach to the development of a permanently manned Mars base. The objectives for a permanently manned Mars base are numerous. Primarily, human presence on Mars will allow utilization of new resources for the improvement of the quality of life on Earth, allowing for new discoveries in technologies, the solar system, and human physiology. Such a mission would also encourage interaction between different countries, increasing international cooperation and leading to a stronger unification of mankind. Surface studies of Mars, scientific experiments in the multiple fields, the research for new minerals, and natural resource production are more immediate goals of the Mars mission. Finally, in the future, colonization of Mars will ensure man's perpetual presence in the universe. Specific objectives of this study were: (1) to design a Mars habitat that minimizes the mass delivered to the Mars surface, provides long-stay capability for the base crew, and accommodates future expansion and modification; (2) to develop a scenario of the construction of a permanently manned Mars base; and (3) to incorporate new and envisioned technologies.

  6. Acid-Base Homeostasis.

    PubMed

    Hamm, L Lee; Nakhoul, Nazih; Hering-Smith, Kathleen S

    2015-12-01

    Acid-base homeostasis and pH regulation are critical for both normal physiology and cell metabolism and function. The importance of this regulation is evidenced by a variety of physiologic derangements that occur when plasma pH is either high or low. The kidneys have the predominant role in regulating the systemic bicarbonate concentration and hence, the metabolic component of acid-base balance. This function of the kidneys has two components: reabsorption of virtually all of the filtered HCO3(-) and production of new bicarbonate to replace that consumed by normal or pathologic acids. This production or generation of new HCO3(-) is done by net acid excretion. Under normal conditions, approximately one-third to one-half of net acid excretion by the kidneys is in the form of titratable acid. The other one-half to two-thirds is the excretion of ammonium. The capacity to excrete ammonium under conditions of acid loads is quantitatively much greater than the capacity to increase titratable acid. Multiple, often redundant pathways and processes exist to regulate these renal functions. Derangements in acid-base homeostasis, however, are common in clinical medicine and can often be related to the systems involved in acid-base transport in the kidneys.

  7. LOGO Based Instruction.

    ERIC Educational Resources Information Center

    Yusuf, Mian Muhammad

    Findings of a 1989 research study on LOGO-based instruction for teaching abstract concepts of geometry are reported in an effort to encourage mathematics teachers to use the LOGO language in computer-assisted instruction. Thirty-three ninth graders (19 males and 14 females) (6 white students and 27 black students) were involved in the controlled…

  8. Polymer based tunneling sensor

    NASA Technical Reports Server (NTRS)

    Cui, Tianhong (Inventor); Wang, Jing (Inventor); Zhao, Yongjun (Inventor)

    2006-01-01

    A process for fabricating a polymer based circuit by the following steps. A mold of a design is formed through a lithography process. The design is transferred to a polymer substrate through a hot embossing process. A metal layer is then deposited over at least part of said design and at least one electrical lead is connected to said metal layer.

  9. Skull Base Tumors

    NASA Astrophysics Data System (ADS)

    Schulz-Ertner, Daniela

    In skull base tumors associated with a low radiosensitivity for conventional radiotherapy (RT), irradiation with proton or carbon ion beams facilitates a safe and accurate application of high tumor doses due to the favorable beam localization properties of these particle beams. Cranial nerves, the brain stem and normal brain tissue can at the same time be optimally spared.

  10. Space base antenna study

    NASA Technical Reports Server (NTRS)

    Deerkoski, L. F.

    1971-01-01

    The field of view required of the space base antenna is defined for both the tracking and data relay satellite link and detached module links. The gain requirements are established and the feasibility of alternative antenna configurations using phased arrays and reflectors are considered. One recommended and one alternative configuration are presented for each of the required links.

  11. Risk Assessment: Evidence Base

    NASA Technical Reports Server (NTRS)

    Johnson-Throop, Kathy A.

    2007-01-01

    Human systems PRA (Probabilistic Risk Assessment: a) Provides quantitative measures of probability, consequence, and uncertainty; and b) Communicates risk and informs decision-making. Human health risks rated highest in ISS PRA are based on 1997 assessment of clinical events in analog operational settings. Much work remains to analyze remaining human health risks identified in Bioastronautics Roadmap.

  12. Problem-Based Learning

    ERIC Educational Resources Information Center

    Allen, Deborah E.; Donham, Richard S.; Bernhardt, Stephen A.

    2011-01-01

    In problem-based learning (PBL), students working in collaborative groups learn by resolving complex, realistic problems under the guidance of faculty. There is some evidence of PBL effectiveness in medical school settings where it began, and there are numerous accounts of PBL implementation in various undergraduate contexts, replete with…

  13. Performance-based ratemaking

    SciTech Connect

    Cross, P.S.

    1995-07-15

    Performance-based ratemaking (PBR) departs from the cost-of-service standard in setting just and reasonable utility rates, but that departure isn`t as easy as it looks. Up until now, cost-of-service ratemaking has provided relatively stable rates, while enabling utilities to attract enormous amounts of capital. Of late, however, regulators appear to be heeding the argument that changing markets warrant a second look. Throughout the country and across the utility industry, some regulators appear willing to abandon cost of service as a proxy for competition, instead favoring performance-based methods that would rely on competitive forces. These performance-based schemes vary in their details but generally afford utilities the opportunity to increase profits by exceeding targets for efficiency and cost savings. Moreover, these plans purport to streamline the regulatory process. Annual, accounting-type reviews replace rate hearings. Cost-of-service studies might not be required at all once initial rates are fixed. Nevertheless, these PBR plans rely on cost-based rates as a starting point and still contain safeguards to protect ratepayers. PBR falls short of true deregulation. As the Massachusetts Department of Public Utilities noted recently in an order approving a PBR variant known as price-cap regulation for New England Telephone and Telegraph Co., `price-cap regulation is not deregulation; it is merely another way for regulators to control the rates charged by a firm.`

  14. School Based Health Centers

    ERIC Educational Resources Information Center

    Children's Aid Society, 2012

    2012-01-01

    School Based Health Centers (SBHC) are considered by experts as one of the most effective and efficient ways to provide preventive health care to children. Few programs are as successful in delivering health care to children at no cost to the patient, and where they are: in school. For many underserved children, The Children's Aid Society's…

  15. Concrete lunar base investigation

    NASA Technical Reports Server (NTRS)

    Lin, T. D.; Senseny, Jonathan A.; Arp, Larry D.; Lindbergh, Charles

    1992-01-01

    This paper presents results of structural analyses and a preliminary design of a precast, prestressed concrete lunar base subjected to 1-atm internal pressure. The proposed infrastructure measures 120 ft in diameter and 72 ft in height, providing 33,000 sq ft of work area for scientific and industrial operations. Three loading conditions were considered in the design (1) during construction, (2) under pressurization, and (3) during an air-leak scenario. A floating foundation, capable of rigid body rotation and translation as the lunar soil beneath it yields, was developed to support the infrastructure and to ensure the airtightness of the system. Results reveal that it is feasible to use precast, prestressed concrete for construction of large lunar bases on the Moon.

  16. Concrete lunar base investigation

    NASA Technical Reports Server (NTRS)

    Lin, T. D.; Senseney, Jonathan A.; Arp, Larry Dean; Lindbergh, Charles

    1989-01-01

    This paper presents results of structural analyses and a preliminary design of a precast, prestressed concrete lunar based subjected to one atmosphere internal pressure. The proposed infrastructure measures 120 ft in diameter and 72 ft in height, providing 33,000 sq ft of work area for scientific and industrial operations. Three loading conditions were considered in the design: (1) during construction; (2) under pressurization; and (3) during an air-leak scenario. A floating foundation, capable of rigid body rotation and translation as the lunar soil beneath it yields, was developed to support the infrastructure and to ensure the air-tightness of the system. Results reveal that it is feasible to use precast, prestressed concrete for construction of large lunar bases on the moon.

  17. WormBase 2007

    PubMed Central

    Rogers, Anthony; Antoshechkin, Igor; Bieri, Tamberlyn; Blasiar, Darin; Bastiani, Carol; Canaran, Payan; Chan, Juancarlos; Chen, Wen J.; Davis, Paul; Fernandes, Jolene; Fiedler, Tristan J.; Han, Michael; Harris, Todd W.; Kishore, Ranjana; Lee, Raymond; McKay, Sheldon; Müller, Hans-Michael; Nakamura, Cecilia; Ozersky, Philip; Petcherski, Andrei; Schindelman, Gary; Schwarz, Erich M.; Spooner, Will; Tuli, Mary Ann; Van Auken, Kimberly; Wang, Daniel; Wang, Xiaodong; Williams, Gary; Yook, Karen; Durbin, Richard; Stein, Lincoln D.; Spieth, John; Sternberg, Paul W.

    2008-01-01

    WormBase (www.wormbase.org) is the major publicly available database of information about Caenorhabditis elegans, an important system for basic biological and biomedical research. Derived from the initial ACeDB database of C. elegans genetic and sequence information, WormBase now includes the genomic, anatomical and functional information about C. elegans, other Caenorhabditis species and other nematodes. As such, it is a crucial resource not only for C. elegans biologists but the larger biomedical and bioinformatics communities. Coverage of core areas of C. elegans biology will allow the biomedical community to make full use of the results of intensive molecular genetic analysis and functional genomic studies of this organism. Improved search and display tools, wider cross-species comparisons and extended ontologies are some of the features that will help scientists extend their research and take advantage of other nematode species genome sequences. PMID:17991679

  18. Space construction data base

    NASA Technical Reports Server (NTRS)

    1979-01-01

    Construction of large systems in space is a technology requiring the development of construction methods to deploy, assemble, and fabricate the elements comprising such systems. A construction method is comprised of all essential functions and operations and related support equipment necessary to accomplish a specific construction task in a particular way. The data base objective is to provide to the designers of large space systems a compendium of the various space construction methods which could have application to their projects.

  19. [Evidence based medicine].

    PubMed

    Cuestas, Eduardo

    2005-01-01

    Evidence based medicine is a systematic method employed to secure the best scientific available evidence when making clinical decisions. Several steps are taken in these process, describing a clinical scenario, formulating a specific clinical question, searching the literature for the pertinent studies, selecting the relevant articles using rules of evidence, understanding and calculating measures of effect, and finally incorporating the evidence and patients preferences in the clinical decision process.

  20. Pavement base drain evaluation

    NASA Astrophysics Data System (ADS)

    Hoffman, G. L.

    1981-06-01

    Portions of a highway drainage system design was revised. Essentially, the longitudinal drainage trench was moved closer to the pavement/shoulder joint, and the fine concrete sand layer was eliminated as a trench backfill material. The specified backfill material is a coarser crushed aggregate (pea gravel). An evaluation of the effects of these changes on pavement performance is given and the new pavement base drain system is compared to the older pipe foundation underdrain system at the same site.

  1. Graphene-based biosensors

    NASA Astrophysics Data System (ADS)

    Lebedev, A. A.; Davydov, V. Yu.; Novikov, S. N.; Litvin, D. P.; Makarov, Yu. N.; Klimovich, V. B.; Samoilovich, M. P.

    2016-07-01

    Results of developing and testing graphene-based sensors capable of detecting protein molecules are presented. The biosensor operation was checked using an immunochemical system comprising fluorescein dye and monoclonal antifluorescein antibodies. The sensor detects fluorescein concentration on a level of 1-10 ng/mL and bovine serum albumin-fluorescein conjugate on a level of 1-5 ng/mL. The proposed device has good prospects for use for early diagnostics of various diseases.

  2. Refrigerator Based on Chemisorption

    NASA Technical Reports Server (NTRS)

    Jones, Jack A.

    1987-01-01

    Reversible chemical reaction generates pressurized oxygen for cooling. Concept for cryogenic refrigerator based on chemical absorption of oxygen by praseodymium/cerium oxide (PCO) compound. Refrigerator produces cryogenic liquid for cooling infrared sensors. Also used for liquefying air and separating oxygen from nitrogen in air. In chemisorption refrigerator, PCO alternately absorbs and desorbs oxygen depending on whether cooled or heated. One pair of compressors accepts oxygen while others releases it. Compressed oxygen liquefied when precooked and expanded.

  3. Flock-based microfluidics.

    PubMed

    Hitzbleck, Martina; Lovchik, Robert D; Delamarche, Emmanuel

    2013-05-21

    Flock-based microfluidics are created by depositing hydrophilic microfibers on an adhesive-coated substrate using an electric field. This enables the fabrication of self-powered microfluidics from one or more different kinds of fibers that form 2D and 3D flowpaths, which can wick 40 microliters of liquid per square centimeter. With this approach, large areas of functional wicking materials can be produced at extremely low cost.

  4. Space Based Communications

    NASA Technical Reports Server (NTRS)

    Simpson, James; Denson, Erik; Valencia, Lisa; Birr, Richard

    2003-01-01

    Current space lift launches on the Eastern and Western Range require extensive ground-based real-time tracking, communications and command/control systems. These are expensive to maintain and operate and cover only limited geographical areas. Future spaceports will require new technologies to provide greater launch and landing opportunities, support simultaneous missions, and offer enhanced decision support models and simulation capabilities. These ranges must also have lower costs and reduced complexity while continuing to provide unsurpassed safety to the public, flight crew, personnel, vehicles and facilities. Commercial and government space-based assets for tracking and communications offer many attractive possibilities to help achieve these goals. This paper describes two NASA proof-of-concept projects that seek-to exploit the advantages of a space-based range: Iridium Flight Modem and Space-Based Telemetry and Range Safety (STARS). Iridium Flight Modem uses the commercial satellite system Iridium for extremely low cost, low rate two-way communications and has been successfully tested on four aircraft flights. A sister project at Goddard Space Flight Center's (GSFC) Wallops Flight Facility (WFF) using the Globalstar system has been tested on one rocket. The basic Iridium Flight Modem system consists of a L1 carrier Coarse/Acquisition (C/A)-Code Global Positioning System (GPS) receiver, an on-board computer, and a standard commercial satellite modem and antennas. STARS uses the much higher data rate NASA owned Tracking and Data Relay Satellite System (TDRSS), a C/A-Code GPS receiver, an experimental low-power transceiver, custom built command and data handler processor, and digitized flight termination system (FTS) commands. STARS is scheduled to fly on an F-15 at Dryden Flight Research Center in the spring of 2003, with follow-on tests over the next several years.

  5. Granular loess classification based

    SciTech Connect

    Browzin, B.S.

    1985-05-01

    This paper discusses how loess might be identified by two index properties: the granulometric composition and the dry unit weight. These two indices are necessary but not always sufficient for identification of loess. On the basis of analyses of samples from three continents, it was concluded that the 0.01-0.5-mm fraction deserves the name loessial fraction. Based on the loessial fraction concept, a granulometric classification of loess is proposed. A triangular chart is used to classify loess.

  6. Evidence-Based Anatomy

    PubMed Central

    Yammine, Kaissar

    2014-01-01

    Anatomy is a descriptive basic medical science that is no longer considered a research-led discipline. Many publications in clinical anatomy are prevalence studies treating clinically relevant anatomical variations and reporting their frequencies and/or associations with variables such as age, sex, side, laterality, and ancestry. This article discusses the need to make sense of the available literature. A new concept, evidence-based anatomy (EBA), is proposed to find, appraise, and synthetize the results reported in such publications. It consists in applying evidence-based principles to the field of epidemiological anatomy research through evidence synthesis using systematic reviews and meta-analyses to generate weighted pooled results. Pooled frequencies and associations based on large pooled sample size are likely to be more accurate and to reflect true population statistics and associations more closely. A checklist of a typical systematic review in anatomy is suggested and the implications of EBA for practice and future research, along with its scope, are discussed. The EBA approach would have positive implications for the future preservation of anatomy as a keystone basic science, for sound knowledge of anatomical variants, and for the safety of medical practice. Clin. Anat. 27:847–852, 2014. PMID:24797314

  7. Incretin-based therapies.

    PubMed

    Neumiller, Joshua J

    2015-01-01

    Incretin-based therapies are steadily gaining clinical popularity, with many more products in the developmental pipeline. Current treatment recommendations incorporate GLP-1 RAs and DPP-4 inhibitors as important agents for consideration in the treatment of T2DM owing to their low hypoglycemia risk, ability to address postprandial hyperglycemia (DPP-4 inhibitors and short-acting GLP-1 RAs), and potential for weight reduction (GLP-1 RAs). These properties may likewise prove advantageous in older adults in whom hypoglycemia is particularly undesirable, although older adults may be more prone to the nausea and vomiting associated with GLP-1 RA therapy. Other safety issues for incretin-based therapies, such as pancreatitis, C-cell hyperplasia, and renal failure, should be considered when choosing an appropriate patient to receive such therapies. Ongoing CV outcome studies will further inform the health care community regarding the CV safety of incretin-based therapies. The availability of both short-acting and long-acting GLP-1 RAs currently allows practitioners to consider individualized blood glucose trends and therapeutic needs when choosing an optimal agent. PMID:25456646

  8. Example based lesion segmentation

    NASA Astrophysics Data System (ADS)

    Roy, Snehashis; He, Qing; Carass, Aaron; Jog, Amod; Cuzzocreo, Jennifer L.; Reich, Daniel S.; Prince, Jerry; Pham, Dzung

    2014-03-01

    Automatic and accurate detection of white matter lesions is a significant step toward understanding the progression of many diseases, like Alzheimer's disease or multiple sclerosis. Multi-modal MR images are often used to segment T2 white matter lesions that can represent regions of demyelination or ischemia. Some automated lesion segmentation methods describe the lesion intensities using generative models, and then classify the lesions with some combination of heuristics and cost minimization. In contrast, we propose a patch-based method, in which lesions are found using examples from an atlas containing multi-modal MR images and corresponding manual delineations of lesions. Patches from subject MR images are matched to patches from the atlas and lesion memberships are found based on patch similarity weights. We experiment on 43 subjects with MS, whose scans show various levels of lesion-load. We demonstrate significant improvement in Dice coefficient and total lesion volume compared to a state of the art model-based lesion segmentation method, indicating more accurate delineation of lesions.

  9. Mobile lunar base project

    NASA Astrophysics Data System (ADS)

    Kozlov, I. A.; Shevchenko, V. V.

    1995-01-01

    An explorer must possess maximal mobility on the Moon if he is to discover natural anomalies most interesting for investigation. The same problem arises in the case of utilization of lunar natural resources. Moreover, according to lunar ecology requirements we should not destroy lunar surface layers over a wide area. For mining processes, many small plots should be chosen far away from each other. The concept of a mobile lunar manned base is proposed. The base structure consists of three vertical cylindrical modules placed into triangular (top view) girder construction. Each module is 5 meters in diameter with a height of 7 meters. The space around the cylinders is filled by a one meter protective layer of lunar soil. The ends of three vertical tube-type supports are put on the separate chassis. Total volume of living and working space is about 350 cubic meters. These modules are sized for a crew of nine. The velocity of the mobile lunar base is about 8 km per hour on a horizontal surface.

  10. AFE base flow computations

    NASA Technical Reports Server (NTRS)

    Venkatapathy, Ethiraj; Prabhu, Dinesh K.; Palmer, Grant

    1991-01-01

    Hypersonic wake flows behind the Aeroassist Flight Experiment (AFE) geometry are analyzed using two Navier-Stokes flow solvers. Many of the AFE wake features observed in ballistic-range shadowgraphs are simulated using a simple, two-dimensional semicylinder geometry at moderate angles of attack. At free-stream conditions corresponding to a Hypersonic Free Flight Facility (HFFF) AFE experiment, the three-dimensional base flow for the AFE geometry is computed using an ideal-gas, Navier-Stokes solver. The computed results agree reasonably well with the shadowgraphs taken at the HFFF. An ideal-gas and a nonequilibrium Navier-Stokes solver have been coupled and applied to the complete flow around the AFE vehicle at the free-stream conditions corresponding to a nomial trajectory point. Limitations of the coupled ideal-gas and nonequilibrium solution are discussed. The nonequilibrium base flow solution is analyzed for the wake radiation and the radiation profiles along various lines of sight are compared. Finally, the wake unsteadiness is predicted using experimental correlations and the numerical solutions. An adaptive grid code, SAGE, has been used in all the simulations to enhance the solution accuracy. The grid adaptation is found to be necessary in obtaining base flow solutions with accurate flow features.

  11. Example Based Lesion Segmentation

    PubMed Central

    Roy, Snehashis; He, Qing; Carass, Aaron; Jog, Amod; Cuzzocreo, Jennifer L.; Reich, Daniel S.; Prince, Jerry; Pham, Dzung

    2016-01-01

    Automatic and accurate detection of white matter lesions is a significant step toward understanding the progression of many diseases, like Alzheimer’s disease or multiple sclerosis. Multi-modal MR images are often used to segment T2 white matter lesions that can represent regions of demyelination or ischemia. Some automated lesion segmentation methods describe the lesion intensities using generative models, and then classify the lesions with some combination of heuristics and cost minimization. In contrast, we propose a patch-based method, in which lesions are found using examples from an atlas containing multi-modal MR images and corresponding manual delineations of lesions. Patches from subject MR images are matched to patches from the atlas and lesion memberships are found based on patch similarity weights. We experiment on 43 subjects with MS, whose scans show various levels of lesion-load. We demonstrate significant improvement in Dice coefficient and total lesion volume compared to a state of the art model-based lesion segmentation method, indicating more accurate delineation of lesions.

  12. RNA based evolutionary optimization.

    PubMed

    Schuster, P

    1993-12-01

    The notion of an RNA world has been introduced for a prebiotic scenario that is dominated by RNA molecules and their properties, in particular their capabilities to act as templates for reproduction and as catalysts for several cleavage and ligation reactions of polynucleotides and polypeptides. This notion is used here also for simple experimental assays which are well suited to study evolution in the test tube. In molecular evolution experiments fitness is determined in essence by the molecular structures of RNA molecules. Evidence is presented for adaptation to environment in cell-free media. RNA based molecular evolution experiments have led to interesting spin-offs in biotechnology, commonly called 'applied molecular evolution', which make use of Darwinian trial-and-error strategies in order to synthesize new pharmacological compounds and other advanced materials on a biological basis. Error-propagation in RNA replication leads to formation of mutant spectra called 'quasispecies'. An increase in the error rate broadens the mutant spectrum. There exists a sharply defined threshold beyond which heredity breaks down and evolutionary adaptation becomes impossible. Almost all RNA viruses studied so far operate at conditions close to this error threshold. Quasispecies and error thresholds are important for an understanding of RNA virus evolution, and they may help to develop novel antiviral strategies. Evolution of RNA molecules can be studied and interpreted by considering secondary structures. The notion of sequence space introduces a distance between pairs of RNA sequences which is tantamount to counting the minimal number of point mutations required to convert the sequences into each other. The mean sensitivity of RNA secondary structures to mutation depends strongly on the base pairing alphabet: structures from sequences which contain only one base pair (GC or AU are much less stable against mutation than those derived from the natural (AUGC) sequences

  13. Lunar base initiative 1992

    NASA Astrophysics Data System (ADS)

    Koelle, H. H.

    The return to the Moon is no longer a question of yes or no, but a question of when and how. The first landing of a human being on the lunar surface in 1969 was a purely national effort of the U.S.A. Building a lunar base and operating it in the next century is rather a task for all nations of this planet, even if one nation could do it alone. However, there are several alternatives to carry out such a program and these will and should be debated during the next years on an urgent basis. To do this, one has to take into account not only the historical accomplishments and the present trends of cooperation in space programs, but also recent geopolitical developments as well as the frame of reference established by international law. The case for an International Lunar Base (ILB) has been presented to the International Academy of Astronautics on 11 October 1987 by the IAA Ad Hoc Committee "Return-to-the-Moon". This draft of a position paper was subsequently published in Acta Astronautica Vol. 17, No. 5, (pp. 463-489) with the request of public debate particularly by the members of the Academy. Some 80 Academicians responded to this invitation by the President of the Academy and voiced their opinions on the questions and issues raised by this draft of a position paper. This led to a refinement of the arguments and assumptions made and it is now possible to prepare an improved position paper proposing concrete steps which may lead to an ILB. An issue of this proportion must start with a discussion of goals and objectives to be arranged in some kind of a ranked order. It also has to take note of the limitations existing at any time by the availability of suitable space transportation systems. These will determine the acquisition date and rate of growth of a lunar base. The logistics system will also greatly influence the base characteristics and layout. The availability of heavy lift launch vehicles would simplify the task and allow to concentrate the construction

  14. DOM Based XSS Detecting Method Based on Phantomjs

    NASA Astrophysics Data System (ADS)

    Dong, Ri-Zhan; Ling, Jie; Liu, Yi

    Because malicious code does not appear in html source code, DOM based XSS cannot be detected by traditional methods. By analyzing the causes of DOM based XSS, this paper proposes a detection method of DOM based XSS based on phantomjs. This paper uses function hijacking to detect dangerous operation and achieves a prototype system. Comparing with existing tools shows that the system improves the detection rate and the method is effective to detect DOM based XSS.

  15. Polyolefin-Based Aerogels

    NASA Technical Reports Server (NTRS)

    Lee, Je Kyun; Gould, George

    2012-01-01

    An organic polybutadiene (PB) rubberbased aerogel insulation material was developed that will provide superior thermal insulation and inherent radiation protection, exhibiting the flexibility, resiliency, toughness, and durability typical of the parent polymer, yet with the low density and superior insulation properties associated with the aerogels. The rubbery behaviors of the PB rubber-based aerogels are able to overcome the weak and brittle nature of conventional inorganic and organic aerogel insulation materials. Additionally, with higher content of hydrogen in their structure, the PB rubber aerogels will also provide inherently better radiation protection than those of inorganic and carbon aerogels. Since PB rubber aerogels also exhibit good hydrophobicity due to their hydrocarbon molecular structure, they will provide better performance reliability and durability as well as simpler, more economic, and environmentally friendly production over the conventional silica or other inorganic-based aerogels, which require chemical treatment to make them hydrophobic. Inorganic aerogels such as silica aerogels demonstrate many unusual and useful properties. There are several strategies to overcoming the drawbacks associated with the weakness and brittleness of silica aerogels. Development of the flexible fiber-reinforced silica aerogel composite blanket has proven one promising approach, providing a conveniently fielded form factor that is relatively robust toward handling in industrial environments compared to silica aerogel monoliths. However, the flexible silica aerogel composites still have a brittle, dusty character that may be undesirable, or even intolerable, in certain applications. Although the cross-linked organic aerogels such as resorcinol-formaldehyde (RF), polyisocyanurate, and cellulose aerogels show very high impact strength, they are also very brittle with little elongation (i.e., less rubbery). Also, silica and carbon aerogels are less efficient

  16. Knowledge based programming at KSC

    NASA Technical Reports Server (NTRS)

    Tulley, J. H., Jr.; Delaune, C. I.

    1986-01-01

    Various KSC knowledge-based systems projects are discussed. The objectives of the knowledge-based automatic test equipment and Shuttle connector analysis network projects are described. It is observed that knowledge-based programs must handle factual and expert knowledge; the characteristics of these two types of knowledge are examined. Applications for the knowledge-based programming technique are considered.

  17. Potential-Based Achievement Goals

    ERIC Educational Resources Information Center

    Elliot, Andrew; Murayama, Kou; Kobeisy, Ahmed; Lichtenfeld, Stephanie

    2015-01-01

    Background: Self-based achievement goals use one's own intrapersonal trajectory as a standard of evaluation, and this intrapersonal trajectory may be grounded in one's past (past-based goals) or one's future potential (potential-based goals). Potential-based goals have been overlooked in the literature to date. Aims: The primary aim of the present…

  18. Characteristics Data Base

    SciTech Connect

    Lewis, E.D.; Moore, R.S. )

    1990-08-01

    The LWR Serial Numbers Database System (SNDB) contains detailed data about individual, historically discharged LWR spent fuel assemblies. This data includes the reactor where used, the year the assemblies were discharged, the pool where they are currently stored, assembly type, burnup, weight, enrichment, and an estimate of their radiological properties. This information is distributed on floppy disks to users in the nuclear industry to assist in planning for the permanent nuclear waste repository. This document describes the design and development of the SNDB. It provides a complete description of the file structures and an outline of the major code modules. It serves as a reference for a programmer maintaining the system, or for others interested in the technical detail of this database. This is the initial version of the SNDB. It contains historical data through December 31, 1987, obtained from the Energy Information Administration (EIA). EIA obtains the data from the utility companies via the RW-859 Survey Form. It evaluates and standardizes the data and distributes the resulting batch level database as a large file on magnetic tape. The Characteristics Data Base obtains this database for use in the LWR Quantities Data Base. Additionally, the CDB obtains the individual assembly level detail from EIA for use in the SNDB. While the Quantities Data Base retains only the level of detail necessary for its reporting, the SNDB does retain and use the batch level data to assist in the identification of a particular assembly serial number. We expect to update the SNDB on an annual basis, as new historical data becomes available.

  19. Internet Based Remote Operations

    NASA Technical Reports Server (NTRS)

    Chamberlain, James

    1999-01-01

    This is the Final Report for the Internet Based Remote Operations Contract, has performed payload operations research support tasks March 1999 through September 1999. These tasks support the GSD goal of developing a secure, inexpensive data, voice, and video mission communications capability between remote payload investigators and the NASA payload operations team in the International Space Station (ISS) era. AZTek has provided feedback from the NASA payload community by utilizing its extensive payload development and operations experience to test and evaluate remote payload operations systems. AZTek has focused on use of the "public Internet" and inexpensive, Commercial-off-the-shelf (COTS) Internet-based tools that would most benefit "small" (e.g., $2 Million or less) payloads and small developers without permanent remote operations facilities. Such projects have limited budgets to support installation and development of high-speed dedicated communications links and high-end, custom ground support equipment and software. The primary conclusions of the study are as follows: (1) The trend of using Internet technology for "live" collaborative applications such as telescience will continue. The GSD-developed data and voice capabilities continued to work well over the "public" Internet during this period. 2. Transmitting multiple voice streams from a voice-conferencing server to a client PC to be mixed and played on the PC is feasible. 3. There are two classes of voice vendors in the market: - Large traditional phone equipment vendors pursuing integration of PSTN with Internet, and Small Internet startups.The key to selecting a vendor will be to find a company sufficiently large and established to provide a base voice-conferencing software product line for the next several years.

  20. Covering all the bases

    SciTech Connect

    Reimers, R.F.; Gross, M.P.

    1996-04-01

    The use of silent discharge plasma technology (SDPT) to treat volatile organic compounds (VOCs) at McClellan Air Force Base during a two-month test is described. SDPT was developed by Los Alamos National Laboratory and licensed to High Mesa Technologies. SDPT was used at McClellan to treat off-gases from air strippers, vapor-extraction systems, and incinerators at flow rates up to 10.4 scfm; destruction and removal efficiencies up to 99.4% were achieved. SDPT can also treat organic liquids by adding a packaged-bed reactor to the first stage system.

  1. Base drive circuit

    DOEpatents

    Lange, Arnold C.

    1995-01-01

    An improved base drive circuit (10) having a level shifter (24) for providing bistable input signals to a pair of non-linear delays (30, 32). The non-linear delays (30, 32) provide gate control to a corresponding pair of field effect transistors (100, 106) through a corresponding pair of buffer components (88, 94). The non-linear delays (30, 32) provide delayed turn-on for each of the field effect transistors (100, 106) while an associated pair of transistors (72, 80) shunt the non-linear delays (30, 32) during turn-off of the associated field effect transistor (100, 106).

  2. Ground based automated telescope

    SciTech Connect

    Colgate, S.A.; Thompson, W.

    1980-01-01

    Recommendation that a ground-based automated telescope of the 2-meter class be built for remote multiuser use as a natural facility. Experience dictates that a primary consideration is a time shared multitasking operating system with virtual memory overlayed with a real time priority interrupt. The primary user facility is a remote terminal networked to the single computer. Many users must have simultaneous time shared access to the computer for program development. The telescope should be rapid slewing, and hence a light weight construction. Automation allows for the closed loop pointing error correction independent of extreme accuracy of the mount.

  3. Mars base buildup scenarios

    SciTech Connect

    Blacic, J.D.

    1985-01-01

    Two surface base build-up scenarios are presented in order to help visualize the mission and to serve as a basis for trade studies. In the first scenario, direct manned landings on the Martian surface occur early in the missions and scientific investigation is the main driver and rationale. In the second scenario, early development of an infrastructure to exploite the volatile resources of the Martian moons for economic purposes is emphasized. Scientific exploration of the surface is delayed at first, but once begun develops rapidly aided by the presence of a permanently manned orbital station.

  4. NICKEL-BASE ALLOY

    DOEpatents

    Inouye, H.; Manly, W.D.; Roche, T.K.

    1960-01-19

    A nickel-base alloy was developed which is particularly useful for the containment of molten fluoride salts in reactors. The alloy is resistant to both salt corrosion and oxidation and may be used at temperatures as high as 1800 deg F. Basically, the alloy consists of 15 to 22 wt.% molybdenum, a small amount of carbon, and 6 to 8 wt.% chromium, the balance being nickel. Up to 4 wt.% of tungsten, tantalum, vanadium, or niobium may be added to strengthen the alloy.

  5. Evidence-based medicine

    PubMed Central

    Shah, Dheeraj; Sachdev, HPS

    2007-01-01

    Evidence based medicine is the practice of solving the clinical problems in one's practice by judicious and systematic use of the medical literature. This includes framing questions rightly and searching the right kind of literature. Thereafter, the available evidence needs to be evaluated for the validity, strength and effect size. Finally, the results are examined for applicability to the current problem which requires a detailed knowledge of the clinical setting, patient profile and the issues related to cost and harm. The present communication deals with these issues in a step-wise manner in order to stimulate readers to practise this important art. PMID:21124675

  6. Mars base buildup scenarios

    NASA Technical Reports Server (NTRS)

    Blacic, J. D.

    1986-01-01

    Two Mars surface based build-up scenarios are presented in order to help visualize the mission and to serve as a basis for trade studies. In the first scenario, direct manned landings on the Martian surface occur early in the missions and scientific investigation is the main driver and rationale. In the second senario, Earth development of an infrastructure to exploit the volatile resources of the Martian moons for economic purposes is emphasized. Scientific exploration of the surface is delayed at first in this scenario relative to the first, but once begun develops rapidly, aided by the presence of a permanently manned orbital station.

  7. Synchrotron based proton drivers

    SciTech Connect

    Weiren Chou

    2002-09-19

    Proton drivers are the proton sources that produce intense short proton bunches. They have a wide range of applications. This paper discusses the proton drivers based on high-intensity proton synchrotrons. It gives a review of the high-intensity proton sources over the world and a brief report on recent developments in this field in the U.S. high-energy physics (HEP) community. The Fermilab Proton Driver is used as a case study for a number of challenging technical design issues.

  8. Internet-Based Communication

    PubMed Central

    Gernsbacher, Morton Ann

    2015-01-01

    Google the question, “How is the Internet changing the way we communicate?,” and you will find no shortage of opinions, or fears, about the Internet altering the way we communicate. Although the Internet is not necessarily making communication briefer (neither is the Internet making communication less formal), the Internet is manifesting our preference for writing over speaking. I propose that our preference for communicating through Internet-based text derives from a fundamental feature of writing: In contrast to speech, which is most often synchronous, text is most often asynchronous. PMID:26330702

  9. Commonality based interoperability

    NASA Astrophysics Data System (ADS)

    Moulton, Christine L.; Hepp, Jared J.; Harrell, John

    2016-05-01

    What interoperability is and why the Army wants it between systems is easily understood. Enabling multiple systems to work together and share data across boundaries in a co-operative manner will benefit the warfighter by allowing for easy access to previously hard-to-reach capabilities. How to achieve interoperability is not as easy to understand due to the numerous different approaches that accomplish the goal. Commonality Based Interoperability (CBI) helps establish how to achieve the goal by extending the existing interoperability definition. CBI is not an implementation, nor is it an architecture; it is a definition of interoperability with a foundation of establishing commonality between systems.

  10. TUNGSTEN BASE ALLOYS

    DOEpatents

    Schell, D.H.; Sheinberg, H.

    1959-12-15

    A high-density quaternary tungsten-base alloy having high mechanical strength and good machinability composed of about 2 wt.% Ni, 3 wt.% Cu, 5 wt.% Pb, and 90wt.% W is described. This alloy can be formed by the powder metallurgy technique of hot pressing in a graphite die without causing a reaction between charge and the die and without formation of a carbide case on the final compact, thereby enabling re-use of the graphite die. The alloy is formable at hot- pressing temperatures of from about 1200 to about 1350 deg C. In addition, there is little component shrinkage, thereby eliminating the necessity of subsequent extensive surface machining.

  11. Base drive circuit

    DOEpatents

    Lange, A.C.

    1995-04-04

    An improved base drive circuit having a level shifter for providing bistable input signals to a pair of non-linear delays. The non-linear delays provide gate control to a corresponding pair of field effect transistors through a corresponding pair of buffer components. The non-linear delays provide delayed turn-on for each of the field effect transistors while an associated pair of transistors shunt the non-linear delays during turn-off of the associated field effect transistor. 2 figures.

  12. Accelerator-based BNCT.

    PubMed

    Kreiner, A J; Baldo, M; Bergueiro, J R; Cartelli, D; Castell, W; Thatar Vento, V; Gomez Asoia, J; Mercuri, D; Padulo, J; Suarez Sandin, J C; Erhardt, J; Kesque, J M; Valda, A A; Debray, M E; Somacal, H R; Igarzabal, M; Minsky, D M; Herrera, M S; Capoulat, M E; Gonzalez, S J; del Grosso, M F; Gagetti, L; Suarez Anzorena, M; Gun, M; Carranza, O

    2014-06-01

    The activity in accelerator development for accelerator-based BNCT (AB-BNCT) both worldwide and in Argentina is described. Projects in Russia, UK, Italy, Japan, Israel, and Argentina to develop AB-BNCT around different types of accelerators are briefly presented. In particular, the present status and recent progress of the Argentine project will be reviewed. The topics will cover: intense ion sources, accelerator tubes, transport of intense beams, beam diagnostics, the (9)Be(d,n) reaction as a possible neutron source, Beam Shaping Assemblies (BSA), a treatment room, and treatment planning in realistic cases.

  13. Accelerator-based BNCT.

    PubMed

    Kreiner, A J; Baldo, M; Bergueiro, J R; Cartelli, D; Castell, W; Thatar Vento, V; Gomez Asoia, J; Mercuri, D; Padulo, J; Suarez Sandin, J C; Erhardt, J; Kesque, J M; Valda, A A; Debray, M E; Somacal, H R; Igarzabal, M; Minsky, D M; Herrera, M S; Capoulat, M E; Gonzalez, S J; del Grosso, M F; Gagetti, L; Suarez Anzorena, M; Gun, M; Carranza, O

    2014-06-01

    The activity in accelerator development for accelerator-based BNCT (AB-BNCT) both worldwide and in Argentina is described. Projects in Russia, UK, Italy, Japan, Israel, and Argentina to develop AB-BNCT around different types of accelerators are briefly presented. In particular, the present status and recent progress of the Argentine project will be reviewed. The topics will cover: intense ion sources, accelerator tubes, transport of intense beams, beam diagnostics, the (9)Be(d,n) reaction as a possible neutron source, Beam Shaping Assemblies (BSA), a treatment room, and treatment planning in realistic cases. PMID:24365468

  14. Sustainability Base Construction Update

    NASA Technical Reports Server (NTRS)

    Mewhinney, Michael

    2012-01-01

    Construction of the new Sustainability Base Collaborative support facility, expected to become the highest performing building in the federal government continues at NASA's Ames Research Center, Moffet Field, Calif. The new building is designed to achieve a platinum rating under the leadership in Energy and Environment Design (LEED) new construction standards for environmentally sustainable construction developed by the U. S. Green Building Council, Washington, D. C. When completed by the end of 2011, the $20.6 million building will feature near zero net energy consumption, use 90 percent less potable water than conventionally build buildings of equivalent size, and will result in reduced building maintenance costs.

  15. Molecular characterization of dichloromethane-degrading Hyphomicrobium strains using 16S rDNA and DCM dehalogenase gene sequences.

    PubMed

    Nikolausz, Marcell; Kappelmeyer, Uwe; Nijenhuis, Ivonne; Ziller, Katja; Kästner, Matthias

    2005-09-01

    A phylogenetic analysis of 6 strains of dichloromethane (DCM) utilizing bacteria was performed. Based on the almost complete 16S rDNA sequence determination, all strains clustered together and showed high sequence similarity to Hyphomicrobium denitrificans, except for the strain MC8b, which is only moderately related to them and probably represents a distinct species. The 16S rDNA-based phylogenetic tree was compared to the one obtained from the DNA sequence data of the dcmA gene coding DCM dehalogenase, the key enzyme of DCM utilization. The topology of the two trees is in good agreement and may suggest an ancient origin of DCM dehalogenase, but also raises questions about the original role of the enzyme. PMID:16156115

  16. Model Based Definition

    NASA Technical Reports Server (NTRS)

    Rowe, Sidney E.

    2010-01-01

    In September 2007, the Engineering Directorate at the Marshall Space Flight Center (MSFC) created the Design System Focus Team (DSFT). MSFC was responsible for the in-house design and development of the Ares 1 Upper Stage and the Engineering Directorate was preparing to deploy a new electronic Configuration Management and Data Management System with the Design Data Management System (DDMS) based upon a Commercial Off The Shelf (COTS) Product Data Management (PDM) System. The DSFT was to establish standardized CAD practices and a new data life cycle for design data. Of special interest here, the design teams were to implement Model Based Definition (MBD) in support of the Upper Stage manufacturing contract. It is noted that this MBD does use partially dimensioned drawings for auxiliary information to the model. The design data lifecycle implemented several new release states to be used prior to formal release that allowed the models to move through a flow of progressive maturity. The DSFT identified some 17 Lessons Learned as outcomes of the standards development, pathfinder deployments and initial application to the Upper Stage design completion. Some of the high value examples are reviewed.

  17. Model-Based Systems

    NASA Technical Reports Server (NTRS)

    Frisch, Harold P.

    2007-01-01

    Engineers, who design systems using text specification documents, focus their work upon the completed system to meet Performance, time and budget goals. Consistency and integrity is difficult to maintain within text documents for a single complex system and more difficult to maintain as several systems are combined into higher-level systems, are maintained over decades, and evolve technically and in performance through updates. This system design approach frequently results in major changes during the system integration and test phase, and in time and budget overruns. Engineers who build system specification documents within a model-based systems environment go a step further and aggregate all of the data. They interrelate all of the data to insure consistency and integrity. After the model is constructed, the various system specification documents are prepared, all from the same database. The consistency and integrity of the model is assured, therefore the consistency and integrity of the various specification documents is insured. This article attempts to define model-based systems relative to such an environment. The intent is to expose the complexity of the enabling problem by outlining what is needed, why it is needed and how needs are being addressed by international standards writing teams.

  18. SPACE BASED INTERCEPTOR SCALING

    SciTech Connect

    G. CANAVAN

    2001-02-01

    Space Based Interceptor (SBI) have ranges that are adequate to address rogue ICBMs. They are not overly sensitive to 30-60 s delay times. Current technologies would support boost phase intercept with about 150 interceptors. Higher acceleration and velocity could reduce than number by about a factor of 3 at the cost of heavier and more expensive Kinetic Kill Vehicles (KKVs). 6g SBI would reduce optimal constellation costs by about 35%; 8g SBI would reduce them another 20%. Interceptor ranges fall rapidly with theater missile range. Constellations increase significantly for ranges under 3,000 km, even with advanced interceptor technology. For distributed launches, these estimates recover earlier strategic scalings, which demonstrate the improved absentee ratio for larger or multiple launch areas. Constellations increase with the number of missiles and the number of interceptors launched at each. The economic estimates above suggest that two SBI per missile with a modest midcourse underlay is appropriate. The SBI KKV technology would appear to be common for space- and surface-based boost phase systems, and could have synergisms with improved midcourse intercept and discrimination systems. While advanced technology could be helpful in reducing costs, particularly for short range theater missiles, current technology appears adequate for pressing rogue ICBM, accidental, and unauthorized launches.

  19. Nanotechnology-Based Cosmeceuticals

    PubMed Central

    Lohani, Alka; Verma, Anurag; Joshi, Himanshi; Yadav, Niti; Karki, Neha

    2014-01-01

    Cosmeceuticals are the fastest growing segment of the personal care industry, and a number of topical cosmeceutical treatments for conditions such as photoaging, hyperpigmentation, wrinkles, and hair damage have come into widespread use. In the cosmeceutical arena nanotechnology has played an important role. Using new techniques to manipulate matter at an atomic or molecular level, they have been at the root of numerous innovations, opening up new perspectives for the future of cosmeceutical industry. Nanotechnology-based cosmeceuticals offer the advantage of diversity in products, and increased bioavailability of active ingredients and increase the aesthetic appeal of cosmeceutical products with prolonged effects. However increased use of nanotechnology in cosmeceuticals has raised concern about the possible penetration of nanoparticles through the skin and potential hazards to the human health. This review outlines the different nanoparticles used in various classes of cosmeceuticals, nanotechnology-based cosmeceutical products present in the market, and the potential risk caused by nanoparticles on exposure and recent regulatory steps taken to overcome them. PMID:24963412

  20. Data base simulator

    SciTech Connect

    Pack, D. J.

    1982-03-01

    This document describes the features of and input to a computer program written for the purpose of generating data bases whose data values contain deterministically known errors. The development of the computer program was motivated by the need to assess automatic data editing procedures for data validation of real data bases. The observed values in the simulated data are the sum of generated true values and generated error values. For a given variable, true data values may be generated by any of the following six methods: frequency distribution, conditional frequency distribution, analysis of variance model, multiple regression model, ARIMA time series model, membership within a defined constrained region. The error values for a given variable may be simulated from an independent distribution or from a distribution dependent upon the error values of other specified variables. The computer program described can be used to satisfy other needs in the area of data simulation beyond the specific need expressed above. Since the addition of errors to the true values is optional, one may readily simulate observed data for variables using one or more of the six previously listed methods.

  1. Soils based productivity evaluation

    SciTech Connect

    Hooks, C.

    1998-12-31

    Since the passage of SMCRA, reclamation success on agricultural lands has been determined by long-term yield testing. This required a long bond release period lasting ten years or more. Recently, landowners, mine operators, and regulators have voiced a need for methods to expedite the bond release process. The financial burdens of annual cropping and field maintenance by mine operators and monitoring by regulators are of major concern. Landowners need to have the land returned to their production operations instead of being locked in the bond release process for a decade or more. A soils based formula could relieve these financial burdens and ensure the most efficient process to return the productive soil resource to the landowner. In addition, this method will also identify problem fields immediately after reclamation. Currently, some fields undergo ten years of testing before a problem becomes evident. Once a problem is identified and remediation occurs, another period of testing is required. A soils based productivity index, currently being developed in Illinois, will be presented. This will include the basic concepts and findings from earlier research. The needs, benefits, and limitations will also be discussed.

  2. Moon base reactor system

    NASA Technical Reports Server (NTRS)

    Chavez, H.; Flores, J.; Nguyen, M.; Carsen, K.

    1989-01-01

    The objective of our reactor design is to supply a lunar-based research facility with 20 MW(e). The fundamental layout of this lunar-based system includes the reactor, power conversion devices, and a radiator. The additional aim of this reactor is a longevity of 12 to 15 years. The reactor is a liquid metal fast breeder that has a breeding ratio very close to 1.0. The geometry of the core is cylindrical. The metallic fuel rods are of beryllium oxide enriched with varying degrees of uranium, with a beryllium core reflector. The liquid metal coolant chosen was natural lithium. After the liquid metal coolant leaves the reactor, it goes directly into the power conversion devices. The power conversion devices are Stirling engines. The heated coolant acts as a hot reservoir to the device. It then enters the radiator to be cooled and reenters the Stirling engine acting as a cold reservoir. The engines' operating fluid is helium, a highly conductive gas. These Stirling engines are hermetically sealed. Although natural lithium produces a lower breeding ratio, it does have a larger temperature range than sodium. It is also corrosive to steel. This is why the container material must be carefully chosen. One option is to use an expensive alloy of cerbium and zirconium. The radiator must be made of a highly conductive material whose melting point temperature is not exceeded in the reactor and whose structural strength can withstand meteor showers.

  3. Ground based infrared astronomy

    NASA Technical Reports Server (NTRS)

    Jennings, D. E.

    1988-01-01

    Infrared spectroscopic instrumentation has been developed for ground-based measurements of astrophysical objects in the intermediate infrared. A conventional Michelson interferometer is limited for astronomical applications in the intermediate infrared by quantum noise fluctuations in the radiation form the source and/or background incident on the detector, and the multiplex advantage is no longer available. One feasible approach to recovering the multiplex advantage is post-dispersion. The infrared signal after passing through telescope and interferometer, is dispersed by a low resolution grating spectrometer onto an array of detectors. The feasibility of the post-dispersion system has been demonstrated with observations of astrophysical objects in the 5 and 10 micrometer atmospheric windows from ground-based telescopes. During FY87/88 the post-disperser was used at the Kitt Peak 4-meter telescope and McMath telescope with facility Fourier transform spectrometers. Jupiter, Saturn, Mars, and Venus were observed. On Jupiter, the resolution at 12 micrometer was 0.01/cm, considerably higher than had been acheived previously. The spectrum contains Jovian ethane and acetylene emission. Construction was begun on the large cryogenic grating spectrometer.

  4. International data base.

    PubMed

    Quick, S

    1984-12-01

    The concept of the International Data Base (IDB) grew from recognition of the need for timely, high quality information on the demographic, social and economic characteristics of foreign countries. During the past 2 decades, the Bureau of the Census, US Department of Commerce, has been compiling, analyzing and evaluating international demographic data and, to a lesser extent, socioeconomic data with particular emphasis on developing countries. In 1979, at the request of the Office of Women in Development at the Agency for International Development, a computerized data base of demographic and socioeconomic statistics that could be used to assess the status of women in developing countries was established. The major categories of data now being included in IDB are as follows: population by age and sex; vital rates, infant mortality and life tables; health and nutrition; fertility and child survivorship; migration; provinces and cities; family planning; ethnic, religious and language groups; literacy and education; labor force, employment, income and gross national product; and household size and housing indicators. The Bureau, through its Center for International Reserch (CIR), developed a computerized central depository of demographic, social and economic data for all countries of the world to serve the needs of the public and private sectors. The initial emphasis was on demographic and social data, since these subject areas are those for which a vast amount of data has already been compiled, analyzed and evaluated by CIR staff. However, meetings are being planned with users to determine the additional types of data that are desired, particularly in the economic and health-related areas. IDB includes data for all countries of the world, by urban and rural residence. The time coverage is from 1950 to the present. Data sources include: population and industrial censuses and surveys, administrative records, population registers, statistical publications, research

  5. Thermoset-Based Nanocomposites

    NASA Astrophysics Data System (ADS)

    Bhembe, Pele

    2002-03-01

    The field of polymer-Clay nanocomposites has attracted considerable attention as a method of enhancing polymer properties and extending their utility, by using molecular or nanoscale reinforcements rather than conventional particulate field microcomposites. Layered silicates dispersed as a reinforcing phase in a polymer matrix are one of the most important forms of such inorganic-organic nanocomposites, making them the subject of intense research. While a significant amount of work has been published on thermoplastic based nanocomposites, however, comparatively few studies of thermoset-based systems have been published. Thus, our research is centered on elucidating the structure-property relationships of thermoset-based nanocomposites. We have developed a series of layered silicate/thermoset nanocomposites using several thermoset polymers (epoxies(di and tetrafunctional), cyanate esters and PMR-15 polyimide). Wide angle X-ray diffraction suggests that intercalated morphologies were obtained for the cases studied. The glass transition temperature has been found to vary as the organic modifier and its amount is varied. For difunctional epoxy samples dispersed with Cloisite 30B, a commercially available nanoclay, the Tg increased by twenty degrees upon addition of as little as 2viscoelastic behavior of these materials has also been investigated using dynamic mechanical analysis. A modest increase in the glassy storage modulus was obtained as the amount of nanoclay increased, with a significant increase in the plateau modulus. Additionally, master curves have been generated using time-temperature superposition, allowing further analysis of the effect of the nanoclay on the relaxation behavior. Activation energies calculated from Arrhenius plots increase as the clay contents increase. These effects will be discussed in the presentation. The fracture toughness increased upon addition of nanoclays while the CTE decreased. Interestingly, the onset of decomposition

  6. MS Based Metabonomics

    SciTech Connect

    Want, Elizabeth J.; Metz, Thomas O.

    2010-03-01

    Metabonomics is the latest and least mature of the systems biology triad, which also includes genomics and proteomics, and has its origins in the early orthomolecular medicine work pioneered by Linus Pauling and Arthur Robinson. It was defined by Nicholson and colleagues in 1999 as the quantitative measurement of perturbations in the metabolite complement of an integrated biological system in response to internal or external stimuli, and is often used today to describe many non-global types of metabolite analyses. Applications of metabonomics are extensive and include toxicology, nutrition, pharmaceutical research and development, physiological monitoring and disease diagnosis. For example, blood samples from millions of neonates are tested routinely by mass spectrometry (MS) as a diagnostic tool for inborn errors of metabolism. The metabonome encompasses a wide range of structurally diverse metabolites; therefore, no single analytical platform will be sufficient. Specialized sample preparation and detection techniques are required, and advances in NMR and MS technologies have led to enhanced metabonome coverage, which in turn demands improved data analysis approaches. The role of MS in metabonomics is still evolving as instrumentation and software becomes more sophisticated and as researchers realize the strengths and limitations of current technology. MS offers a wide dynamic range, high sensitivity, and reproducible, quantitative analysis. These attributes are essential for addressing the challenges of metabonomics, as the range of metabolite concentrations easily exceeds nine orders of magnitude in biofluids, and the diversity of molecular species ranges from simple amino and organic acids to lipids and complex carbohydrates. Additional challenges arise in generating a comprehensive metabolite profile, downstream data processing and analysis, and structural characterization of important metabolites. A typical workflow of MS-based metabonomics is shown in Figure

  7. Information-based clustering

    PubMed Central

    Slonim, Noam; Atwal, Gurinder Singh; Tkačik, Gašper; Bialek, William

    2005-01-01

    In an age of increasingly large data sets, investigators in many different disciplines have turned to clustering as a tool for data analysis and exploration. Existing clustering methods, however, typically depend on several nontrivial assumptions about the structure of data. Here, we reformulate the clustering problem from an information theoretic perspective that avoids many of these assumptions. In particular, our formulation obviates the need for defining a cluster “prototype,” does not require an a priori similarity metric, is invariant to changes in the representation of the data, and naturally captures nonlinear relations. We apply this approach to different domains and find that it consistently produces clusters that are more coherent than those extracted by existing algorithms. Finally, our approach provides a way of clustering based on collective notions of similarity rather than the traditional pairwise measures. PMID:16352721

  8. Fiber based optofluidic biosensors

    NASA Astrophysics Data System (ADS)

    Lismont, M.; Vandewalle, N.; Joris, B.; Dreesen, L.

    2014-09-01

    Medicinal diagnosis requires the development of innovative devices allowing the detection of small amounts of biological species. Among the large variety of available biosensors, the ones based on fluorescence phenomenon are really promising. Here, we show a prototype of the basic unit of a multi-sensing biosensor combining optics and microfluidics benefits. This unit makes use of two crossed optical fibers: the first fiber is used to carry small probe molecules droplets and excite fluorescence, while the second one is devoted to target molecules droplets transport and fluorescence detection. Within this scheme, the interaction takes place in each fiber node. The main benefits of this detection setup are the absence of fibers functionalization, the use of microliter volumes of target and probe species, their separation before interaction, and a better detection limit compared to cuvettes setups.

  9. Liaison based assembly design

    SciTech Connect

    Ames, A.; Kholwadwala, D.; Wilson, R.H.

    1996-12-01

    Liaison Based Assembly Design extends the current information infrastructure to support design in terms of kinematic relationships between parts, or liaisons. These liaisons capture information regarding contact, degrees-of-freedom constraints and containment relationships between parts in an assembly. The project involved defining a useful collection of liaison representations, investigating their properties, and providing for maximum use of the data in downstream applications. We tested our ideas by implementing a prototype system involving extensions to Pro/Engineer and the Archimedes assembly planner. With an expanded product model, the design system is more able to capture design intent. When a product update is attempted, increased knowledge availability improves our ability to understand the effect of design changes. Manufacturing and analysis disciplines benefit from having liaison information available, so less time is wasted arguing over incomplete design specifications and our enterprise can be more completely integrated.

  10. Institutionally based videoconferencing.

    PubMed

    Caudill, Robert Lee; Sager, Zachary

    2015-01-01

    The delivery of psychiatric care via video-teleconferencing (VTC) technology is thought to have reached a tipping point. As a medical speciality with relatively few material or technical requirements for service delivery, psychiatry has been one of the earliest to embrace the possibility of providing evaluations and treatment at a distance. Such technical infrastructure as is necessary can often be found in the institutions already in existence. It was natural therefore that institutionally based telepsychiatry would lay the foundation for the development of the field. In this article we review the history and development of institutional VTC in a wide variety of clinically supervised settings such as hospitals, outpatient clinics, and forensic settings. We cite evidence supporting institutionally sponsored use and expand on key takeaways for the development and expansion of videoconferencing in these settings. We also speculate on the future direction and development of psychiatric care provided by these arrangements. PMID:26507786

  11. Base isolation: Fresh insight

    SciTech Connect

    Shustov, V.

    1993-07-15

    The objective of the research is a further development of the engineering concept of seismic isolation. Neglecting the transient stage of seismic loading results in a widespread misjudgement: The force of resistance associated with velocity is mostly conceived as a source of damping vibrations, though it is an active force at the same time, during an earthquake type excitation. For very pliant systems such as base isolated structures with relatively low bearing stiffness and with artificially added heavy damping mechanism, the so called `damping`` force may occur even the main pushing force at an earthquake. Thus, one of the two basic pillars of the common seismic isolation philosophy, namely, the doctrine of usefulness and necessity of a strong damping mechanism, is turning out to be a self-deception, sometimes even jeopardizing the safety of structures and discrediting the very idea of seismic isolation. There is a way out: breaking with damping dependancy.

  12. Covariant mutually unbiased bases

    NASA Astrophysics Data System (ADS)

    Carmeli, Claudio; Schultz, Jussi; Toigo, Alessandro

    2016-06-01

    The connection between maximal sets of mutually unbiased bases (MUBs) in a prime-power dimensional Hilbert space and finite phase-space geometries is well known. In this article, we classify MUBs according to their degree of covariance with respect to the natural symmetries of a finite phase-space, which are the group of its affine symplectic transformations. We prove that there exist maximal sets of MUBs that are covariant with respect to the full group only in odd prime-power dimensional spaces, and in this case, their equivalence class is actually unique. Despite this limitation, we show that in dimension 2r covariance can still be achieved by restricting to proper subgroups of the symplectic group, that constitute the finite analogues of the oscillator group. For these subgroups, we explicitly construct the unitary operators yielding the covariance.

  13. Endoscopic Skull Base Surgery

    PubMed Central

    Senior, Brent A

    2008-01-01

    Endoscopic skull base surgery has undergone rapid advancement in the past decade moving from pituitary surgery to suprasellar lesions and now to a myriad of lesions extending from the cribriform plate to C2 and laterally out to the infratemporal fossa and petrous apex. Evolution of several technological advances as well as advances in understanding of endoscopic anatomy and the development of surgical techniques both in resection and reconstruction have fostered this capability. Management of benign disease via endoscopic methods is largely accepted now but more data is needed before the controversy on the role of endoscopic management of malignant disease is decided. Continued advances in surgical technique, navigation systems, endoscopic imaging technology, and robotics assure continued brisk evolution in this expanding field. PMID:19434274

  14. Surface Plasmon Based Spectrometer

    NASA Astrophysics Data System (ADS)

    Wig, Andrew; Passian, Ali; Boudreaux, Philip; Ferrell, Tom

    2008-03-01

    A spectrometer that uses surface plasmon excitation in thin metal films to separate light into its component wavelengths is described. The use of surface plasmons as a dispersive medium sets this spectrometer apart from prism, grating, and interference based variants and allows for the miniaturization of this device. Theoretical and experimental results are presented for two different operation models. In the first case surface plasmon tunneling in the near field is used to provide transmission spectra of different broad band-pass, glass filters across the visible wavelength range with high stray-light rejection at low resolution as well as absorption spectra of chlorophyll extracted from a spinach leaf. The second model looks at the far field components of surface plasmon scattering.

  15. Race-based therapeutics.

    PubMed

    Yancy, Clyde W

    2008-08-01

    The issue of race in medicine is problematic. Race is not a physiologic grouping, and all persons of a given race do not necessarily share the same clinical phenotype or genetic substrate. Despite clear signals that certain risk factors and diseases vary as a function of race, translating those differences into race-based therapeutics has been awkward and has done little to change the natural history of cardiovascular disease as it affects special populations. Among the varied special populations, the African American population appears to have the most significant and adverse variances for cardiovascular disease as well as worrisome signals that drug responsiveness varies. Recent guideline statements have now acknowledged certain treatment options that are most appropriate for African Americans with cardiovascular disease, especially hypertension and heart failure. As more physiologic markers of disease and drug responsiveness become available, the need for racial designations in medicine may lessen, and therapies can be optimized for all patients without regard to race or ethnicity.

  16. 3. SOUTH SIDE. Edwards Air Force Base, South Base ...

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

    3. SOUTH SIDE. - Edwards Air Force Base, South Base Sled Track, Firing & Control Blockhouse for 10,000-foot Track, South of Sled Track at midpoint of 20,000-foot track, Lancaster, Los Angeles County, CA

  17. DNA-based machines.

    PubMed

    Wang, Fuan; Willner, Bilha; Willner, Itamar

    2014-01-01

    The base sequence in nucleic acids encodes substantial structural and functional information into the biopolymer. This encoded information provides the basis for the tailoring and assembly of DNA machines. A DNA machine is defined as a molecular device that exhibits the following fundamental features. (1) It performs a fuel-driven mechanical process that mimics macroscopic machines. (2) The mechanical process requires an energy input, "fuel." (3) The mechanical operation is accompanied by an energy consumption process that leads to "waste products." (4) The cyclic operation of the DNA devices, involves the use of "fuel" and "anti-fuel" ingredients. A variety of DNA-based machines are described, including the construction of "tweezers," "walkers," "robots," "cranes," "transporters," "springs," "gears," and interlocked cyclic DNA structures acting as reconfigurable catenanes, rotaxanes, and rotors. Different "fuels", such as nucleic acid strands, pH (H⁺/OH⁻), metal ions, and light, are used to trigger the mechanical functions of the DNA devices. The operation of the devices in solution and on surfaces is described, and a variety of optical, electrical, and photoelectrochemical methods to follow the operations of the DNA machines are presented. We further address the possible applications of DNA machines and the future perspectives of molecular DNA devices. These include the application of DNA machines as functional structures for the construction of logic gates and computing, for the programmed organization of metallic nanoparticle structures and the control of plasmonic properties, and for controlling chemical transformations by DNA machines. We further discuss the future applications of DNA machines for intracellular sensing, controlling intracellular metabolic pathways, and the use of the functional nanostructures for drug delivery and medical applications.

  18. Electrical Percolation Based Biosensors

    PubMed Central

    Bruck, Hugh Alan; Yang, Minghui; Kostov, Yordan; Rasooly, Avraham

    2013-01-01

    A new approach to label free biosensing has been developed based on the principle of “electrical percolation”. In electrical percolation, long-range electrical connectivity is formed in randomly oriented and distributed systems of discrete elements. By applying this principle to biological interactions, it is possible to measure biological components both directly and electronically. The main element for electrical percolation biosensor is the biological semiconductor (BSC) which is a multi-layer 3-D carbon nanotube-antibody network. In the BSC, molecular interactions, such as binding of antigens to the antibodies, disrupt the network continuity causing increased resistance of the network. BSCs can be fabricated by immobilizing conducting elements, such as pre-functionalized single-walled carbon nanotubes (SWNTs)-antibody complex, directly onto a substrate, such as a Poly(methyl methacrylate) (PMMA) surface (also known as plexi-glass or Acrylic). BSCs have been demonstrated for direct (label-free) electronic measurements of antibody-antigen binding using SWNTs. If the concentration of the SWNT network is slightly above the electrical percolation threshold, then binding of a specific antigen to the pre-functionalized SWNT dramatically increases the electrical resistance due to changes in the tunneling between the SWNTs. Using anti-Staphylococcal enterotoxin B (SEB) IgG as a “gate” and SEB as an “actuator”, it was demonstrated that the BSC was able to detect SEB at concentrations of 1 ng/ml. Based on this concept, an automated configuration for BSCs is described here that enables real time continuous detection. The new BSC configuration may permit assembly of multiple sensors on the same chip to create “Biological Central Processing Units (CPUs)” with multiple biological elements, capable of processing and sorting out information on multiple analytes simultaneously. PMID:24041756

  19. DNA-based machines.

    PubMed

    Wang, Fuan; Willner, Bilha; Willner, Itamar

    2014-01-01

    The base sequence in nucleic acids encodes substantial structural and functional information into the biopolymer. This encoded information provides the basis for the tailoring and assembly of DNA machines. A DNA machine is defined as a molecular device that exhibits the following fundamental features. (1) It performs a fuel-driven mechanical process that mimics macroscopic machines. (2) The mechanical process requires an energy input, "fuel." (3) The mechanical operation is accompanied by an energy consumption process that leads to "waste products." (4) The cyclic operation of the DNA devices, involves the use of "fuel" and "anti-fuel" ingredients. A variety of DNA-based machines are described, including the construction of "tweezers," "walkers," "robots," "cranes," "transporters," "springs," "gears," and interlocked cyclic DNA structures acting as reconfigurable catenanes, rotaxanes, and rotors. Different "fuels", such as nucleic acid strands, pH (H⁺/OH⁻), metal ions, and light, are used to trigger the mechanical functions of the DNA devices. The operation of the devices in solution and on surfaces is described, and a variety of optical, electrical, and photoelectrochemical methods to follow the operations of the DNA machines are presented. We further address the possible applications of DNA machines and the future perspectives of molecular DNA devices. These include the application of DNA machines as functional structures for the construction of logic gates and computing, for the programmed organization of metallic nanoparticle structures and the control of plasmonic properties, and for controlling chemical transformations by DNA machines. We further discuss the future applications of DNA machines for intracellular sensing, controlling intracellular metabolic pathways, and the use of the functional nanostructures for drug delivery and medical applications. PMID:24647836

  20. Base Research Program

    SciTech Connect

    Everett Sondreal; John Hendrikson

    2009-03-31

    In June 2009, the Energy & Environmental Research Center (EERC) completed 11 years of research under the U.S. Department of Energy (DOE) Base Cooperative Agreement No. DE-FC26-98FT40320 funded through the Office of Fossil Energy (OFE) and administered at the National Energy Technology Laboratory (NETL). A wide range of diverse research activities were performed under annual program plans approved by NETL in seven major task areas: (1) resource characterization and waste management, (2) air quality assessment and control, (3) advanced power systems, (4) advanced fuel forms, (5) value-added coproducts, (6) advanced materials, and (7) strategic studies. This report summarizes results of the 67 research subtasks and an additional 50 strategic studies. Selected highlights in the executive summary illustrate the contribution of the research to the energy industry in areas not adequately addressed by the private sector alone. During the period of performance of the agreement, concerns have mounted over the impact of carbon emissions on climate change, and new programs have been initiated by DOE to ensure that fossil fuel resources along with renewable resources can continue to supply the nation's transportation fuel and electric power. The agreement has addressed DOE goals for reductions in CO{sub 2} emissions through efficiency, capture, and sequestration while expanding the supply and use of domestic energy resources for energy security. It has further contributed to goals for near-zero emissions from highly efficient coal-fired power plants; environmental control capabilities for SO{sub 2}, NO{sub x}, fine respirable particulate (PM{sub 2.5}), and mercury; alternative transportation fuels including liquid synfuels and hydrogen; and synergistic integration of fossil and renewable resources (e.g., wind-, biomass-, and coal-based electrical generation).

  1. Loyalty-based management.

    PubMed

    Reichheld, F F

    1993-01-01

    Despite a flurry of activities aimed at serving customers better, few companies have systematically revamped their operations with customer loyalty in mind. Instead, most have adopted improvement programs ad hoc, and paybacks haven't materialized. Building a highly loyal customer base must be integral to a company's basic business strategy. Loyalty leaders like MBNA credit cards are successful because they have designed their entire business systems around customer loyalty--a self-reinforcing system in which the company delivers superior value consistently and reinvents cash flows to find and keep high-quality customers and employees. The economic benefits of high customer loyalty are measurable. When a company consistently delivers superior value and wins customer loyalty, market share and revenues go up, and the cost of acquiring new customers goes down. The better economics mean the company can pay workers better, which sets off a whole chain of events. Increased pay boosts employee moral and commitment; as employees stay longer, their productivity goes up and training costs fall; employees' overall job satisfaction, combined with their experience, helps them serve customers better; and customers are then more inclined to stay loyal to the company. Finally, as the best customers and employees become part of the loyalty-based system, competitors are left to survive with less desirable customers and less talented employees. To compete on loyalty, a company must understand the relationships between customer retention and the other parts of the business--and be able to quantify the linkages between loyalty and profits. It involves rethinking and aligning four important aspects of the business: customers, product/service offering, employees, and measurement systems.

  2. ASCOT data base management system

    SciTech Connect

    Barbieri, J.; Nyholm, R.; Castro, C.; Hill, K.

    1980-07-01

    The ASCOT data base management system is designed to handle the data produced by both the experimental and theoretical efforts of the DOE Atmospheric Studies in Complex Terrain (ASCOT) project. The data base envisioned is hierarchically structured, sparse, and compact. Information concerning any given data file is stored in a directory file. The data base management system uses a relational data management approach. Presently three management schema are being developed for use with the data base. 5 figures.

  3. Verification of knowledge bases based on containment checking

    SciTech Connect

    Levy. A.Y.; Rousset, M.C.

    1996-12-31

    Building complex knowledge based applications requires encoding large amounts of domain knowledge. After acquiring knowledge from domain experts, much of the effort in building a knowledge base goes into verifying that the knowledge is encoded correctly. We consider the problem of verifying hybrid knowledge bases that contain both Horn rules and a terminology in a description logic. Our approach to the verification problem is based on showing a close relationship to the problem of query containment. Our first contribution, based on this relationship, is presenting a thorough analysis of the decidability and complexity of the verification problem, for knowledge bases containing recursive rules and the interpreted predicates =, {le}, < and {ne}. Second, we show that important new classes of constraints on correct inputs and outputs can be expressed in a hybrid setting, in which a description logic class hierarchy is also considered, and we present the first complete algorithm for verifying such hybrid knowledge bases.

  4. Bio-based backsheet

    NASA Astrophysics Data System (ADS)

    Levy, Stanley B.

    2008-08-01

    A primary goal of Photovoltaics is to generate electricity while reducing reliance on the world's petroleum supply. However, PV backsheets are produced from petro-based chemicals, which, to a certain extent, defeat the purpose of using solar energy. Materials from three sustainable resources were targeted for PV backsheet development: PLA made from corn, a cellulosic made from cotton, and a type of nylon made from castor beans. Some of these films were coated with various materials to lower the WVTR. Modules produced using these backsheets were subjected to rigorous testing, including the damp heat test and the wet Hypot test as outlined in UL 1703. As cast PLA film tends to be very brittle. This problem is solved with additives or biaxial orientation. PLA film is UV stable and highly transparent which would merit it for consideration as a front glazing as well as for a backsheet. However, its moisture resistance is not robust. A cellulosic film made from cotton was considered which has a continuous duty temperature rating of 105°C. This product had to be modified significantly to convert it from a hydrophilic film to a hydrophobic one. Additionally, this material has an RTI value of 90°C. Nylon 11, produced from castor beans, is very interesting because it is bio-sustainable, but not biodegradable. It has improved moisture properties over the more common nylons, and has an RTI value of 105°C.

  5. Lunar base construction requirements

    NASA Technical Reports Server (NTRS)

    Jolly, Steve; Helleckson, Brent

    1990-01-01

    The following viewgraph presentation is a review of the Lunar Base Constructibility Study carried out in the spring and summer of 1990. The objective of the study was to develop a method for evaluating the constructibility of Phase A proposals to build facilities on orbit or on extraterrestrial surfaces. Space construction was broadly defined as all forms of assembly, disassembly, connection, disconnection, deployment, stowage, excavation, emplacement, activation, test, transportation, etc., required to create facilities in orbit and on the surfaces of other celestial bodies. It was discovered that decisions made in the face of stated and unstated assumptions early in the design process (commonly called Phase A) can lock in non-optimal construction methods. Often, in order to construct the design, alterations must be made to the design during much later phases of the project. Such 'fixes' can be very difficult, expensive, or perhaps impossible. Assessing constructibility should thus be a part of the iterative design process, starting with the Phase A studies and continuing through production. This study assumes that there exists a minimum set of key construction requirements (i.e., questions whose answers form the set of discriminators) that must be implied or specified in order to assess the constructibility of the design. This set of construction requirements constitutes a 'constructibility filter' which then becomes part of the iterative design process. Five inherently different, dichotomous design reference missions were used in the extraction of these requirements to assure the depth and breath of the list.

  6. Alphavirus-based vaccines.

    PubMed

    Lundstrom, Kenneth

    2014-06-01

    Alphavirus vectors have demonstrated high levels of transient heterologous gene expression both in vitro and in vivo and, therefore, possess attractive features for vaccine development. The most commonly used delivery vectors are based on three single-stranded encapsulated alphaviruses, namely Semliki Forest virus, Sindbis virus and Venezuelan equine encephalitis virus. Alphavirus vectors have been applied as replication-deficient recombinant viral particles and, more recently, as replication-proficient particles. Moreover, in vitro transcribed RNA, as well as layered DNA vectors have been applied for immunization. A large number of highly immunogenic viral structural proteins expressed from alphavirus vectors have elicited strong neutralizing antibody responses in multispecies animal models. Furthermore, immunization studies have demonstrated robust protection against challenges with lethal doses of virus in rodents and primates. Similarly, vaccination with alphavirus vectors expressing tumor antigens resulted in prophylactic protection against challenges with tumor-inducing cancerous cells. As certain alphaviruses, such as Chikungunya virus, have been associated with epidemics in animals and humans, attention has also been paid to the development of vaccines against alphaviruses themselves. Recent progress in alphavirus vector development and vaccine technology has allowed conducting clinical trials in humans.

  7. Lunar base heat pump

    NASA Technical Reports Server (NTRS)

    Goldman, Jeffrey H.; Tetreault, R.; Fischbach, D.; Walker, D.

    1994-01-01

    A heat pump is a device which elevates the temperature of a heat flow by a means of an energy input. By doing this, the heat pump can cause heat to transfer faster from a warm region to a cool region, or it can cause heat to flow from a cool region to a warmer region. The second case is the one which finds vast commercial applications such as air conditioning, heating, and refrigeration. Aerospace applications of heat pumps include both cases. The NASA Johnson Space Center is currently developing a Life Support Systems Integration Facility (LSSIF, previously SIRF) to provide system-level integration, operational test experience, and performance data that will enable NASA to develop flight-certified hardware for future planetary missions. A high lift heat pump is a significant part of the TCS hardware development associated with the LSSIF. The high lift heat pump program discussed here is being performed in three phases. In Phase 1, the objective is to develop heat pump concepts for a lunar base, a lunar lander, and for a ground development unit for the SIRF. In Phase 2, the design of the SIRF ground test unit is being performed, including identification and evaluation of safety and reliability issues. In Phase 3, the SIRF unit will be manufactured, tested, and delivered to the NASA Johnson Space Center.

  8. Mentalization-Based Treatment

    PubMed Central

    Bateman, Anthony; Fonagy, Peter

    2013-01-01

    The concept of mentalizing has captured the interest and imagination of an astonishing range of people—from psychoanalysts to neuroscientists, from child development researchers to geneticists, from existential philosophers to phenomenologists—all of whom seem to have found it useful. According to the Thompson Reuter maintained Web of Science, the use of the term in titles and abstracts of scientific papers increased from 10 to 2,750 between 1991 and 2011. Clinicians in particular have enthusiastically embraced the idea, and have put it to innovative use in their practices. Mentalization-based treatment (MBT)—making mentalizing a core focus of therapy—was initially developed for the treatment of borderline personality disorder (BPD) in routine clinical services delivered in group and individual modalities. Therapy with mentalizing as a central component is currently being developed for treatment of numerous groups, including people with antisocial personality disorder, substance abuse, eating disorders, and at-risk mothers with infants and children (A. Bateman & Fonagy, 2011). It is also being used with families and adolescents, in schools, and in managing social groups (Asen & Fonagy, 2011; Fonagy et al., 2009; Twemlow, Fonagy, & Sacco, 2005a, 2005b). In this article, we focus on MBT in the treatment of BPD. PMID:26157198

  9. Constraint-based scheduling

    NASA Technical Reports Server (NTRS)

    Zweben, Monte

    1991-01-01

    The GERRY scheduling system developed by NASA Ames with assistance from the Lockheed Space Operations Company, and the Lockheed Artificial Intelligence Center, uses a method called constraint based iterative repair. Using this technique, one encodes both hard rules and preference criteria into data structures called constraints. GERRY repeatedly attempts to improve schedules by seeking repairs for violated constraints. The system provides a general scheduling framework which is being tested on two NASA applications. The larger of the two is the Space Shuttle Ground Processing problem which entails the scheduling of all inspection, repair, and maintenance tasks required to prepare the orbiter for flight. The other application involves power allocations for the NASA Ames wind tunnels. Here the system will be used to schedule wind tunnel tests with the goal of minimizing power costs. In this paper, we describe the GERRY system and its applications to the Space Shuttle problem. We also speculate as to how the system would be used for manufacturing, transportation, and military problems.

  10. Evidence-based management.

    PubMed

    Pfeffer, Jeffrey; Sutton, Robert I

    2006-01-01

    For the most part, managers looking to cure their organizational ills rely on obsolete knowledge they picked up in school, long-standing but never proven traditions, patterns gleaned from experience, methods they happen to be skilled in applying, and information from vendors. They could learn a thing or two from practitioners of evidence-based medicine, a movement that has taken the medical establishment by storm over the past decade. A growing number of physicians are eschewing the usual, flawed resources and are instead identifying, disseminating, and applying research that is soundly conducted and clinically relevant. It's time for managers to do the same. The challenge is, quite simply, to ground decisions in the latest and best knowledge of what actually works. In some ways, that's more difficult to do in business than in medicine. The evidence is weaker in business; almost anyone can (and many people do) claim to be a management expert; and a motley crew of sources--Shakespeare, Billy Graham,Jack Welch, Attila the Hunare used to generate management advice. Still, it makes sense that when managers act on better logic and strong evidence, their companies will beat the competition. Like medicine, management is learned through practice and experience. Yet managers (like doctors) can practice their craft more effectively if they relentlessly seek new knowledge and insight, from both inside and outside their companies, so they can keep updating their assumptions, skills, and knowledge.

  11. Nanowire-based detector

    DOEpatents

    Berggren, Karl K; Hu, Xiaolong; Masciarelli, Daniele

    2014-06-24

    Systems, articles, and methods are provided related to nanowire-based detectors, which can be used for light detection in, for example, single-photon detectors. In one aspect, a variety of detectors are provided, for example one including an electrically superconductive nanowire or nanowires constructed and arranged to interact with photons to produce a detectable signal. In another aspect, fabrication methods are provided, including techniques to precisely reproduce patterns in subsequently formed layers of material using a relatively small number of fabrication steps. By precisely reproducing patterns in multiple material layers, one can form electrically insulating materials and electrically conductive materials in shapes such that incoming photons are redirected toward a nearby electrically superconductive materials (e.g., electrically superconductive nanowire(s)). For example, one or more resonance structures (e.g., comprising an electrically insulating material), which can trap electromagnetic radiation within its boundaries, can be positioned proximate the nanowire(s). The resonance structure can include, at its boundaries, electrically conductive material positioned proximate the electrically superconductive nanowire such that light that would otherwise be transmitted through the sensor is redirected toward the nanowire(s) and detected. In addition, electrically conductive material can be positioned proximate the electrically superconductive nanowire (e.g. at the aperture of the resonant structure), such that light is directed by scattering from this structure into the nanowire.

  12. Microsphere based saliva diagnostics

    NASA Astrophysics Data System (ADS)

    Rissin, David M.; DiCesare, Christopher; Hayman, Ryan B.; Blicharz, Timothy M.; Walt, David R.

    2005-11-01

    Saliva presents a minimally invasive alternative medium to blood for performing diagnostics1. Microsphere sensors for ions, small organic molecules, and proteins are currently being developed and optical microarrays containing thousands of these sensors will be used for simultaneous multi-analyte analysis. The fiber bundle platform in use is 1mm in diameter and contains approximately 50,000 individually addressable 3.1μm fibers, each with an etched well capable of housing a single 3.1μm microsphere sensor. Micron-sized bead-based chemistries are produced in house, followed by deposition onto a fiber-optic bundle platform, allowing for multiplexed analysis. The ultimate goal is to develop a universal diagnostic system using saliva as the diagnostic medium. This platform will permit multiplexed analysis of a sample by integrating microfluidics with the optical arrays loaded with sensors capable of detecting relevant biomarkers associated with a wide range of disease states. Disease states that are currently under investigation include end stage renal disease (ESRD) and Sjoegrens Syndrome (SS).

  13. Holography based super resolution

    NASA Astrophysics Data System (ADS)

    Hussain, Anwar; Mudassar, Asloob A.

    2012-05-01

    This paper describes the simulation of a simple technique of superresolution based on holographic imaging in spectral domain. The input beam assembly containing 25 optical fibers with different orientations and positions is placed to illuminate the object in the 4f optical system. The position and orientation of each fiber is calculated with respect to the central fiber in the array. The positions and orientations of the fibers are related to the shift of object spectrum at aperture plane. During the imaging process each fiber is operated once in the whole procedure to illuminate the input object transparency which gives shift to the object spectrum in the spectral domain. This shift of the spectrum is equal to the integral multiple of the pass band aperture width. During the operation of single fiber (ON-state) all other fibers are in OFF-state at that time. The hologram recorded by each fiber at the CCD plane is stored in computer memory. At the end of illumination process total 25 holograms are recorded by the whole fiber array and by applying some post processing and specific algorithm single super resolved image is obtained. The superresolved image is five times better than the band-limited image. The work is demonstrated using computer simulation only.

  14. Scintillator based beta batteries

    NASA Astrophysics Data System (ADS)

    Rensing, Noa M.; Tiernan, Timothy C.; Shirwadkar, Urmila; O'Dougherty, Patrick; Freed, Sara; Hawrami, Rastgo; Squillante, Michael R.

    2013-05-01

    Some long-term, remote applications do not have access to conventional harvestable energy in the form of solar radiation (or other ambient light), wind, environmental vibration, or wave motion. Radiation Monitoring Devices, Inc. (RMD) is carrying out research to address the most challenging applications that need power for many months or years and which have undependable or no access to environmental energy. Radioisotopes are an attractive candidate for this energy source, as they can offer a very high energy density combined with a long lifetime. Both large scale nuclear power plants and radiothermal generators are based on converting nuclear energy to heat, but do not scale well to small sizes. Furthermore, thermo-mechanical power plants depend on moving parts, and RTG's suffer from low efficiency. To address the need for compact nuclear power devices, RMD is developing a novel beta battery, in which the beta emissions from a radioisotope are converted to visible light in a scintillator and then the visible light is converted to electrical power in a photodiode. By incorporating 90Sr into the scintillator SrI2 and coupling the material to a wavelength-matched solar cell, we will create a scalable, compact power source capable of supplying milliwatts to several watts of power over a period of up to 30 years. We will present the latest results of radiation damage studies and materials processing development efforts, and discuss how these factors interact to set the operating life and energy density of the device.

  15. Constraint-based scheduling

    NASA Technical Reports Server (NTRS)

    Zweben, Monte

    1991-01-01

    The GERRY scheduling system developed by NASA Ames with assistance from the Lockheed Space Operations Company, and the Lockheed Artificial Intelligence Center, uses a method called constraint-based iterative repair. Using this technique, one encodes both hard rules and preference criteria into data structures called constraints. GERRY repeatedly attempts to improve schedules by seeking repairs for violated constraints. The system provides a general scheduling framework which is being tested on two NASA applications. The larger of the two is the Space Shuttle Ground Processing problem which entails the scheduling of all the inspection, repair, and maintenance tasks required to prepare the orbiter for flight. The other application involves power allocation for the NASA Ames wind tunnels. Here the system will be used to schedule wind tunnel tests with the goal of minimizing power costs. In this paper, we describe the GERRY system and its application to the Space Shuttle problem. We also speculate as to how the system would be used for manufacturing, transportation, and military problems.

  16. Graphene based GHz detectors

    NASA Astrophysics Data System (ADS)

    Boyd, Anthony K.; El Fatimy, Abdel; Barbara, Paola; Nath, Anindya; Campbell, Paul M.; Myers-Ward, Rachael; Daniels, Kevin; Gaskill, D. Kurt

    Graphene demonstrates great promise as a detector over a wide spectral range especially in the GHz range. This is because absorption is enhanced due to the Drude contribution. In the GHz range there are viable detection mechanisms for graphene devices. With this in mind, two types of GHz detectors are fabricated on epitaxial graphene using a lift off resist-based clean lithography process to produce low contact resistance. Both device types use asymmetry for detection, consistent with recent thoughts of the photothermoelectric effect (PTE) mechanism. The first is an antenna coupled device. It utilizes two dissimilar contact metals and the work function difference produces the asymmetry. The other device is a field effect transistor constructed with an asymmetric top gate that creates a PN junction and facilitates tuning the photovoltaic response. The response of both device types, tested from 100GHz to 170GHz, are reported. This work was sponsored by the U.S. Office of Naval Research (Award Number N000141310865).

  17. Nanomembrane-based plasmonics

    NASA Astrophysics Data System (ADS)

    Jakšić, Zoran; Vuković, Slobodan M.; Buha, Jelena; Matovic, Jovan

    2011-01-01

    This paper reviews the main properties and applications of nanomembrane-based plasmonic structures, including some results presented here for the first time. Artificial nanomembranes are a novel building block in micro- and nanosystems technologies. They represent quasi-two-dimensional (2D) freestanding structures thinner than 100 nm and with giant aspect ratios that often exceed 1,000,000. They may be fabricated as various quasi-2D metal-dielectric nanocomposites with tailorable properties; they are fully symmetric in an electromagnetic sense and support long-range surface plasmon polaritons. This makes nanomembranes a convenient platform for different plasmonic structures such as subwavelength plasmonic crystals and metamaterials and applications such as plasmon waveguides and ultrasensitive bio/chemical sensors. Among other advantages of nanomembrane plasmonics is the feasibility to fabricate flexible, transferable plasmonic guides applicable to different substrates and dynamically tunable through stretching. There are various approaches to multifunctionalization of nanomembranes for plasmonics, including the use of transparent conductive oxide nanoparticles, but also the incorporation of switchable ion channels. Since the natural counterpart of the artificial nanomembranes are cell membranes, the multifunctionalization of synthetic nanomembranes ensures the introduction of bionic principles into plasmonics, at the same time extending the toolbox of the available nanostructures, materials and functions.

  18. Constraint-based scheduling

    NASA Technical Reports Server (NTRS)

    Zweben, Monte

    1993-01-01

    The GERRY scheduling system developed by NASA Ames with assistance from the Lockheed Space Operations Company, and the Lockheed Artificial Intelligence Center, uses a method called constraint-based iterative repair. Using this technique, one encodes both hard rules and preference criteria into data structures called constraints. GERRY repeatedly attempts to improve schedules by seeking repairs for violated constraints. The system provides a general scheduling framework which is being tested on two NASA applications. The larger of the two is the Space Shuttle Ground Processing problem which entails the scheduling of all the inspection, repair, and maintenance tasks required to prepare the orbiter for flight. The other application involves power allocation for the NASA Ames wind tunnels. Here the system will be used to schedule wind tunnel tests with the goal of minimizing power costs. In this paper, we describe the GERRY system and its application to the Space Shuttle problem. We also speculate as to how the system would be used for manufacturing, transportation, and military problems.

  19. Image Data Bases on Campus.

    ERIC Educational Resources Information Center

    Kaplan, Reid; Mathieson, Gordon

    1989-01-01

    A description of how image database technology was used to develop two prototypes for academic and administrative applications at Yale University, one using a video data base integration and the other using document-scanning data base technology, is presented. Technical underpinnings for the creation of data bases are described. (Author/MLW)

  20. PMIS: Data Base Design Report

    ERIC Educational Resources Information Center

    Fiddleman, Richard; Gorman, Michael M.

    1972-01-01

    PMIS is a computer-based planning and management information system for local school districts. This report centers on the PMIS data bases that contain school system data by reviewing the major phases involved in their creation, explaining the factors that caused the unique orientation of the data bases, reviewing the two tasks that comprise the…

  1. Performance-Based Funding Brief

    ERIC Educational Resources Information Center

    Washington Higher Education Coordinating Board, 2011

    2011-01-01

    A number of states have made progress in implementing performance-based funding (PFB) and accountability. This policy brief summarizes main features of performance-based funding systems in three states: Tennessee, Ohio, and Indiana. The brief also identifies key issues that states considering performance-based funding must address, as well as…

  2. Graphene based gene transfection

    NASA Astrophysics Data System (ADS)

    Feng, Liangzhu; Zhang, Shuai; Liu, Zhuang

    2011-03-01

    Graphene as a star in materials research has been attracting tremendous attentions in the past few years in various fields including biomedicine. In this work, for the first time we successfully use graphene as a non-toxic nano-vehicle for efficient gene transfection. Graphene oxide (GO) is bound with cationic polymers, polyethyleneimine (PEI) with two different molecular weights at 1.2 kDa and 10 kDa, forming GO-PEI-1.2k and GO-PEG-10k complexes, respectively, both of which are stable in physiological solutions. Cellular toxicity tests reveal that our GO-PEI-10k complex exhibits significantly reduced toxicity to the treated cells compared to the bare PEI-10k polymer. The positively charged GO-PEI complexes are able to further bind with plasmid DNA (pDNA) for intracellular transfection of the enhanced green fluorescence protein (EGFP) gene in HeLa cells. While EGFP transfection with PEI-1.2k appears to be ineffective, high EGFP expression is observed using the corresponding GO-PEI-1.2k as the transfection agent. On the other hand, GO-PEI-10k shows similar EGFP transfection efficiency but lower toxicity compared with PEI-10k. Our results suggest graphene to be a novel gene delivery nano-vector with low cytotoxicity and high transfection efficiency, promising for future applications in non-viral based gene therapy.Graphene as a star in materials research has been attracting tremendous attentions in the past few years in various fields including biomedicine. In this work, for the first time we successfully use graphene as a non-toxic nano-vehicle for efficient gene transfection. Graphene oxide (GO) is bound with cationic polymers, polyethyleneimine (PEI) with two different molecular weights at 1.2 kDa and 10 kDa, forming GO-PEI-1.2k and GO-PEG-10k complexes, respectively, both of which are stable in physiological solutions. Cellular toxicity tests reveal that our GO-PEI-10k complex exhibits significantly reduced toxicity to the treated cells compared to the bare PEI

  3. Base Excision Repair and Cancer

    PubMed Central

    Wallace, Susan S.; Murphy, Drew L.; Sweasy, Joann B.

    2012-01-01

    Base excision repair is the system used from bacteria to man to remove the tens of thousands of endogenous DNA damages produced daily in each human cell. Base excision repair is required for normal mammalian development and defects have been associated with neurological disorders and cancer. In this paper we provide an overview of short patch base excision repair in humans and summarize current knowledge of defects in base excision repair in mouse models and functional studies on short patch base excision repair germ line polymorphisms and their relationship to cancer. The biallelic germ line mutations that result in MUTYH-associated colon cancer are also discussed. PMID:22252118

  4. Military Bases and Conservation Markets

    SciTech Connect

    Mark Ankeny

    2007-09-01

    Over time, DoD is likely to be one of the largest buyers and sellers in a water quality trading market. The Department of Defense (DoD) operates military bases that resemble small cities in infrastructure. As units redeploy, bases are likely to find themselves well within their environmental limits at the originating base and potentially bumping against limits such as nitrate and phosphate loading at the destination base. Stricter rules and heavier loadings in growing watersheds also present challenges to local bases and municipalities as regulators clamp down on loadings from existing Waste Water Treatment Plants (WWTPs) to meet water quality standards.

  5. Telemetry-Based Ranging

    NASA Technical Reports Server (NTRS)

    Hamkins, Jon; Vilnrotter, Victor A.; Andrews, Kenneth S.; Shambayati, Shervin

    2011-01-01

    A telemetry-based ranging scheme was developed in which the downlink ranging signal is eliminated, and the range is computed directly from the downlink telemetry signal. This is the first Deep Space Network (DSN) ranging technology that does not require the spacecraft to transmit a separate ranging signal. By contrast, the evolutionary ranging techniques used over the years by NASA missions, including sequential ranging (transmission of a sequence of sinusoids) and PN-ranging (transmission of a pseudo-noise sequence) whether regenerative (spacecraft acquires, then regenerates and retransmits a noise-free ranging signal) or transparent (spacecraft feeds the noisy demodulated uplink ranging signal into the downlink phase modulator) relied on spacecraft power and bandwidth to transmit an explicit ranging signal. The state of the art in ranging is described in an emerging CCSDS (Consultative Committee for Space Data Systems) standard, in which a pseudo-noise (PN) sequence is transmitted from the ground to the spacecraft, acquired onboard, and the PN sequence is coherently retransmitted back to the ground, where a delay measurement is made between the uplink and downlink signals. In this work, the telemetry signal is aligned with the uplink PN code epoch. The ground station computes the delay between the uplink signal transmission and the received downlink telemetry. Such a computation is feasible because symbol synchronizability is already an integral part of the telemetry design. Under existing technology, the telemetry signal cannot be used for ranging because its arrival-time information is not coherent with any Earth reference signal. By introducing this coherence, and performing joint telemetry detection and arrival-time estimation on the ground, a high-rate telemetry signal can provide all the precision necessary for spacecraft ranging.

  6. Ferrocene-Based Nanoelectronics

    SciTech Connect

    Lawrence R. Sita

    2006-02-15

    Ferrocene-based molecular components for nanoelectronics offer a number of distinct advantages relative to all carbon frameworks due to metal-centered molecular states that should be closer in energy to the Fermi levels of the metal electrodes in metal / molecule / metal heterojunctions. Given this, the overall goal of the project was to investigate the conduction physics of a variety of proposed ferrocene diode / transistor designs in order to address the fundamental question; can electron transport within nm-length scale structures be modulated in a controlled fashion? During the funded period, substantial progress towards achieving this goal was made by surmounting a number of scientific and technical obstacles. More specifically, a concise and general synthetic route to several mono- and diferrocene dithiols and monothiols was achieved that now allows for the directed and controlled assembly of a variety of metal / molecule /metal test structures for the single molecule conductance measurements and the fabrication of self-assembled monolayers (SAMs) on Au(111) that are amenable to quantitative electrochemical characterization of electron-transfer rates. Most importantly, by using an electromigrated test structure, reproducible I/V data for one of the ferrocene dithiol molecules have been collected which exhibit surprisingly high conductance. Exceptional agreement of this result with theory serves to substantiate the original hypothesis that metal-centered states within a molecular bridge can indeed serve to establish higher conductance relative to all-organic molecular bridges. Overall, the successful demonstration of the ability of ferrocene-molecular frameworks to serve as exceptional molecular conductors will play an important role in the continued evolution in design of molecular components for nanoelectronic devices, which in turn, will have a positive impact on the science and potential technologies associated with these systems.

  7. Space-based detectors

    NASA Astrophysics Data System (ADS)

    Sesana, A.; Weber, W. J.; Killow, C. J.; Perreur-Lloyd, M.; Robertson, D. I.; Ward, H.; Fitzsimons, E. D.; Bryant, J.; Cruise, A. M.; Dixon, G.; Hoyland, D.; Smith, D.; Bogenstahl, J.; McNamara, P. W.; Gerndt, R.; Flatscher, R.; Hechenblaikner, G.; Hewitson, M.; Gerberding, O.; Barke, S.; Brause, N.; Bykov, I.; Danzmann, K.; Enggaard, A.; Gianolio, A.; Vendt Hansen, T.; Heinzel, G.; Hornstrup, A.; Jennrich, O.; Kullmann, J.; Møller-Pedersen, S.; Rasmussen, T.; Reiche, J.; Sodnik, Z.; Suess, M.; Armano, M.; Sumner, T.; Bender, P. L.; Akutsu, T.; Sathyaprakash, B. S.

    2014-12-01

    The parallel session C5 on Space-Based Detectors gave a broad overview over the planned space missions related to gravitational wave detection. Overviews of the revolutionary science to be expected from LISA was given by Alberto Sesana and Sasha Buchman. The launch of LISA Pathfinder (LPF) is planned for 2015. This mission and its payload "LISA Technology Package" will demonstrate key technologies for LISA. In this context, reference masses in free fall for LISA, and gravitational physics in general, was described by William Weber, laser interferometry at the pico-metre level and the optical bench of LPF was presented by Christian Killow and the performance of the LPF optical metrology system by Paul McNamara. While LPF will not yet be sensitive to gravitational waves, it may nevertheless be used to explore fundamental physics questions, which was discussed by Michele Armano. Some parts of the LISA technology that are not going to be demonstrated by LPF, but under intensive development at the moment, were presented by Oliver Jennrich and Oliver Gerberding. Looking into the future, Japan is studying the design of a mid-frequency detector called DECIGO, which was discussed by Tomotada Akutsu. Using atom interferometry for gravitational wave detection has also been recently proposed, and it was critically reviewed by Peter Bender. In the nearer future, the launch of GRACE Follow-On (for Earth gravity observation) is scheduled for 2017, and it will include a Laser Ranging Interferometer as technology demonstrator. This will be the first inter-spacecraft laser interferometer and has many aspects in common with the LISA long arm, as discussed by Andrew Sutton.

  8. Soy-based renoprotection

    PubMed Central

    McGraw, Nancy J; Krul, Elaine S; Grunz-Borgmann, Elizabeth; Parrish, Alan R

    2016-01-01

    Chronic kidney disease (CKD) is a significant public health problem as risk factors such as advanced age, obesity, hypertension and diabetes rise in the global population. Currently there are no effective pharmacologic treatments for this disease. The role of diet is important for slowing the progression of CKD and managing symptoms in later stages of renal insufficiency. While low protein diets are generally recommended, maintaining adequate levels of intake is critical for health. There is an increasing appreciation that the source of protein may also be important. Soybean protein has been the most extensively studied plant-based protein in subjects with kidney disease and has demonstrated renal protective properties in a number of clinical studies. Soy protein consumption has been shown to slow the decline in estimated glomerular filtration rate and significantly improve proteinuria in diabetic and non-diabetic patients with nephropathy. Soy’s beneficial effects on renal function may also result from its impact on certain physiological risk factors for CKD such as dyslipidemia, hypertension and hyperglycemia. Soy intake is also associated with improvements in antioxidant status and systemic inflammation in early and late stage CKD patients. Studies conducted in animal models have helped to identify the underlying molecular mechanisms that may play a role in the positive effects of soy protein on renal parameters in polycystic kidney disease, metabolically-induced kidney dysfunction and age-associated progressive nephropathy. Despite the established relationship between soy and renoprotection, further studies are needed for a clear understanding of the role of the cellular and molecular target(s) of soy protein in maintaining renal function. PMID:27152261

  9. Lunar Base Heat Pump

    NASA Technical Reports Server (NTRS)

    Walker, D.; Fischbach, D.; Tetreault, R.

    1996-01-01

    The objective of this project was to investigate the feasibility of constructing a heat pump suitable for use as a heat rejection device in applications such as a lunar base. In this situation, direct heat rejection through the use of radiators is not possible at a temperature suitable for lde support systems. Initial analysis of a heat pump of this type called for a temperature lift of approximately 378 deg. K, which is considerably higher than is commonly called for in HVAC and refrigeration applications where heat pumps are most often employed. Also because of the variation of the rejection temperature (from 100 to 381 deg. K), extreme flexibility in the configuration and operation of the heat pump is required. A three-stage compression cycle using a refrigerant such as CFC-11 or HCFC-123 was formulated with operation possible with one, two or three stages of compression. Also, to meet the redundancy requirements, compression was divided up over multiple compressors in each stage. A control scheme was devised that allowed these multiple compressors to be operated as required so that the heat pump could perform with variable heat loads and rejection conditions. A prototype heat pump was designed and constructed to investigate the key elements of the high-lift heat pump concept. Control software was written and implemented in the prototype to allow fully automatic operation. The heat pump was capable of operation over a wide range of rejection temperatures and cooling loads, while maintaining cooling water temperature well within the required specification of 40 deg. C +/- 1.7 deg. C. This performance was verified through testing.

  10. Soy-based renoprotection.

    PubMed

    McGraw, Nancy J; Krul, Elaine S; Grunz-Borgmann, Elizabeth; Parrish, Alan R

    2016-05-01

    Chronic kidney disease (CKD) is a significant public health problem as risk factors such as advanced age, obesity, hypertension and diabetes rise in the global population. Currently there are no effective pharmacologic treatments for this disease. The role of diet is important for slowing the progression of CKD and managing symptoms in later stages of renal insufficiency. While low protein diets are generally recommended, maintaining adequate levels of intake is critical for health. There is an increasing appreciation that the source of protein may also be important. Soybean protein has been the most extensively studied plant-based protein in subjects with kidney disease and has demonstrated renal protective properties in a number of clinical studies. Soy protein consumption has been shown to slow the decline in estimated glomerular filtration rate and significantly improve proteinuria in diabetic and non-diabetic patients with nephropathy. Soy's beneficial effects on renal function may also result from its impact on certain physiological risk factors for CKD such as dyslipidemia, hypertension and hyperglycemia. Soy intake is also associated with improvements in antioxidant status and systemic inflammation in early and late stage CKD patients. Studies conducted in animal models have helped to identify the underlying molecular mechanisms that may play a role in the positive effects of soy protein on renal parameters in polycystic kidney disease, metabolically-induced kidney dysfunction and age-associated progressive nephropathy. Despite the established relationship between soy and renoprotection, further studies are needed for a clear understanding of the role of the cellular and molecular target(s) of soy protein in maintaining renal function.

  11. Carbon Nanotube Based Light Sensor

    NASA Technical Reports Server (NTRS)

    Wincheski, russell A. (Inventor); Smits, Jan M. (Inventor); Jordan, Jeffrey D. (Inventor); Watkins, Anthony Neal (Inventor); Ingram, JoAnne L. (Inventor)

    2006-01-01

    A light sensor substrate comprises a base made from a semi-conductive material and topped with a layer of an electrically non-conductive material. A first electrode and a plurality of carbon nanotube (CNT)-based conductors are positioned on the layer of electrically non-conductive material with the CNT-based conductors being distributed in a spaced apart fashion about a periphery of the first electrode. Each CNT-based conductor is coupled on one end thereof to the first electrode and extends away from the first electrode to terminate at a second free end. A second or gate electrode is positioned on the non-conductive material layer and is spaced apart from the second free end of each CNT-based conductor. Coupled to the first and second electrode is a device for detecting electron transfer along the CNT-based conductors resulting from light impinging on the CNT-based conductors.

  12. Case-based reasoning: The marriage of knowledge base and data base

    NASA Technical Reports Server (NTRS)

    Pulaski, Kirt; Casadaban, Cyprian

    1988-01-01

    The coupling of data and knowledge has a synergistic effect when building an intelligent data base. The goal is to integrate the data and knowledge almost to the point of indistinguishability, permitting them to be used interchangeably. Examples given in this paper suggest that Case-Based Reasoning is a more integrated way to link data and knowledge than pure rule-based reasoning.

  13. Potential Role of Nitrite for Abiotic Fe(II) Oxidation and Cell Encrustation during Nitrate Reduction by Denitrifying Bacteria

    PubMed Central

    Klueglein, Nicole; Zeitvogel, Fabian; Stierhof, York-Dieter; Floetenmeyer, Matthias; Konhauser, Kurt O.; Obst, Martin

    2014-01-01

    Microorganisms have been observed to oxidize Fe(II) at neutral pH under anoxic and microoxic conditions. While most of the mixotrophic nitrate-reducing Fe(II)-oxidizing bacteria become encrusted with Fe(III)-rich minerals, photoautotrophic and microaerophilic Fe(II) oxidizers avoid cell encrustation. The Fe(II) oxidation mechanisms and the reasons for encrustation remain largely unresolved. Here we used cultivation-based methods and electron microscopy to compare two previously described nitrate-reducing Fe(II) oxidizers ( Acidovorax sp. strain BoFeN1 and Pseudogulbenkiania sp. strain 2002) and two heterotrophic nitrate reducers (Paracoccus denitrificans ATCC 19367 and P. denitrificans Pd 1222). All four strains oxidized ∼8 mM Fe(II) within 5 days in the presence of 5 mM acetate and accumulated nitrite (maximum concentrations of 0.8 to 1.0 mM) in the culture media. Iron(III) minerals, mainly goethite, formed and precipitated extracellularly in close proximity to the cell surface. Interestingly, mineral formation was also observed within the periplasm and cytoplasm; intracellular mineralization is expected to be physiologically disadvantageous, yet acetate consumption continued to be observed even at an advanced stage of Fe(II) oxidation. Extracellular polymeric substances (EPS) were detected by lectin staining with fluorescence microscopy, particularly in the presence of Fe(II), suggesting that EPS production is a response to Fe(II) toxicity or a strategy to decrease encrustation. Based on the data presented here, we propose a nitrite-driven, indirect mechanism of cell encrustation whereby nitrite forms during heterotrophic denitrification and abiotically oxidizes Fe(II). This work adds to the known assemblage of Fe(II)-oxidizing bacteria in nature and complicates our ability to delineate microbial Fe(II) oxidation in ancient microbes preserved as fossils in the geological record. PMID:24271182

  14. Base pairing and base mis-pairing in nucleic acids

    NASA Technical Reports Server (NTRS)

    Wang, A. H. J.; Rich, A.

    1986-01-01

    In recent years we have learned that DNA is conformationally active. It can exist in a number of different stable conformations including both right-handed and left-handed forms. Using single crystal X-ray diffraction analysis we are able to discover not only additional conformations of the nucleic acids but also different types of hydrogen bonded base-base interactions. Although Watson-Crick base pairings are the predominant type of interaction in double helical DNA, they are not the only types. Recently, we have been able to examine mismatching of guanine-thymine base pairs in left-handed Z-DNA at atomic resolution (1A). A minimum amount of distortion of the sugar phosphate backbone is found in the G x T pairing in which the bases are held together by two hydrogen bonds in the wobble pairing interaction. Because of the high resolution of the analysis we can visualize water molecules which fill in to accommodate the other hydrogen bonding positions in the bases which are not used in the base-base interactions. Studies on other DNA oligomers have revealed that other types of non-Watson-Crick hydrogen bonding interactions can occur. In the structure of a DNA octamer with the sequence d(GCGTACGC) complexed to an antibiotic triostin A, it was found that the two central AT base pairs are held together by Hoogsteen rather than Watson-Crick base pairs. Similarly, the G x C base pairs at the ends are also Hoogsteen rather than Watson-Crick pairing. Hoogsteen base pairs make a modified helix which is distinct from the Watson-Crick double helix.

  15. XML-Based SHINE Knowledge Base Interchange Language

    NASA Technical Reports Server (NTRS)

    James, Mark; Mackey, Ryan; Tikidjian, Raffi

    2008-01-01

    The SHINE Knowledge Base Interchange Language software has been designed to more efficiently send new knowledge bases to spacecraft that have been embedded with the Spacecraft Health Inference Engine (SHINE) tool. The intention of the behavioral model is to capture most of the information generally associated with a spacecraft functional model, while specifically addressing the needs of execution within SHINE and Livingstone. As such, it has some constructs that are based on one or the other.

  16. Computer-based and web-based radiation safety training

    SciTech Connect

    Owen, C., LLNL

    1998-03-01

    The traditional approach to delivering radiation safety training has been to provide a stand-up lecture of the topic, with the possible aid of video, and to repeat the same material periodically. New approaches to meeting training requirements are needed to address the advent of flexible work hours and telecommuting, and to better accommodate individuals learning at their own pace. Computer- based and web-based radiation safety training can provide this alternative. Computer-based and web- based training is an interactive form of learning that the student controls, resulting in enhanced and focused learning at a time most often chosen by the student.

  17. Game-based versus traditional case-based learning

    PubMed Central

    Telner, Deanna; Bujas-Bobanovic, Maja; Chan, David; Chester, Bob; Marlow, Bernard; Meuser, James; Rothman, Arthur; Harvey, Bart

    2010-01-01

    ABSTRACT OBJECTIVE To evaluate family physicians’ enjoyment of and knowledge gained from game-based learning, compared with traditional case-based learning, in a continuing medical education (CME) event on stroke prevention and management. DESIGN An equivalence trial to determine if game-based learning was as effective as case-based learning in terms of attained knowledge levels. Game questions and small group cases were developed. Participants were randomized to either a game-based or a case-based group and took part in the event. SETTING Ontario provincial family medicine conference. PARTICIPANTS Thirty-two family physicians and 3 senior family medicine residents attending the conference. INTERVENTION Participation in either a game-based or a case-based CME learning group. MAIN OUTCOME MEASURES Scores on 40-item immediate and 3-month posttests of knowledge and a satisfaction survey. RESULTS Results from knowledge testing immediately after the event and 3 months later showed no significant difference in scoring between groups. Participants in the game-based group reported higher levels of satisfaction with the learning experience. CONCLUSION Games provide a novel way of organizing CME events. They might provide more group interaction and discussion, as well as improve recruitment to CME events. They might also provide a forum for interdisciplinary CME. Using games in future CME events appears to be a promising approach to facilitate participant learning. PMID:20841574

  18. Lead-based paint assessment

    SciTech Connect

    Lorie, C.; Cowdery, J.W.

    1994-12-31

    In 1977, The US consumer product safety commission banned the use of lead-based paint (LBP) in all industries, except the maritime industry which still has certain privileged uses. Unfortunately for property and building owners, the ban did not come soon enough. In response to this heightened awareness, several environmental market sectors addressing the issues have emerged. These include: residential; soil; commercial; water; and structures. The first and most important step in addressing the concerns posed by the existence of lead based contamination is to quantify the amount of lead-based product, to determine the location of the lead based product and the extent, if any, of lead based contamination, and to make recommendations for the remediation or abatement of the lead product and resultant contamination. In narrowing the focus of these issues, this paper discusses lead-based paint assessment; preparing and organizing the assessment, the regulatory considerations, assessment methodology, and presentation of results.

  19. Concurrent array-based queue

    SciTech Connect

    Heidelberger, Philip; Steinmacher-Burow, Burkhard

    2015-01-06

    According to one embodiment, a method for implementing an array-based queue in memory of a memory system that includes a controller includes configuring, in the memory, metadata of the array-based queue. The configuring comprises defining, in metadata, an array start location in the memory for the array-based queue, defining, in the metadata, an array size for the array-based queue, defining, in the metadata, a queue top for the array-based queue and defining, in the metadata, a queue bottom for the array-based queue. The method also includes the controller serving a request for an operation on the queue, the request providing the location in the memory of the metadata of the queue.

  20. dBASE IV basics

    SciTech Connect

    O`Connor, P.

    1994-09-01

    This is a user`s manual for dBASE IV. dBASE IV is a popular software application that can be used on your personal computer to help organize and maintain your database files. It is actually a set of tools with which you can create, organize, select and manipulate data in a simple yet effective manner. dBASE IV offers three methods of working with the product: (1) control center: (2) command line; and (3) programming.

  1. Principles of models based engineering

    SciTech Connect

    Dolin, R.M.; Hefele, J.

    1996-11-01

    This report describes a Models Based Engineering (MBE) philosophy and implementation strategy that has been developed at Los Alamos National Laboratory`s Center for Advanced Engineering Technology. A major theme in this discussion is that models based engineering is an information management technology enabling the development of information driven engineering. Unlike other information management technologies, models based engineering encompasses the breadth of engineering information, from design intent through product definition to consumer application.

  2. Evidence-Based Medicine: Rhinoplasty.

    PubMed

    Lee, Matthew K; Most, Sam P

    2015-08-01

    Evidence-based medicine has become increasingly prominent in the climate of modern day healthcare. The practice of evidence-based medicine involves the integration of the best available evidence with clinical experience and expertise to help guide clinical decision-making. The essential tenets of evidence-based medicine can be applied to both functional and aesthetic rhinoplasty. Current outcome measures in functional and aesthetic rhinoplasty, including objective, subjective, and clinician-reported measures, is summarized and the current data is reviewed.

  3. Predictability of sacral base levelness based on iliac crest measurements.

    PubMed

    Dott, G A; Hart, C L; McKay, C

    1994-05-01

    A level sacral base plane is necessary to allow normalization of complex lumbosacral mechanics. Palpatory examinations are often used to evaluate for leg length discrepancy and pelvic obliquity despite improved accuracy and consistency of radiographic techniques. Treatment based on palpatory examinations suppose a direct and consistent relationship between the pelvic bones (innominates) and the sacral base. To evaluate the relationship between iliac crest levelness and sacral base levelness, a radiographic postural survey in the upright, weight-bearing position was performed on 358 men and women thought to have pelvic obliquity. Of these subjects, 293 demonstrated unlevel iliac crest heights or sacral base > or = 3/16 inch (4.76 mm), with iliac crest heights accurately predicting sacral base position 62% of the time. At > or = 3/8 inch (9.53 mm), 68% of the cases were accurately predicted. When the criterion for unlevelness was increased to > or = 1/2 inch (12.70 mm), the predictive accuracy improved to 83%. Radiographic findings in this study demonstrate a significant difference between iliac crest heights and sacral base position. In cases of mild to moderate short leg syndromes, the iliac crest height is an unreliable predictor of the direction or degree of sacral base levelness. PMID:8056627

  4. Flavobacterium daejeonense sp. nov. and Flavobacterium suncheonense sp. nov., isolated from greenhouse soils in Korea.

    PubMed

    Kim, Byung-Yong; Weon, Hang-Yeon; Cousin, Sylvie; Yoo, Seung-Hee; Kwon, Soon-Wo; Go, Seung-Joo; Stackebrandt, Erko

    2006-07-01

    Two yellow-pigmented, Gram-negative, rod-shaped bacterial strains, GH1-10(T) and GH29-5(T), were isolated from greenhouse soils in Korea. 16S rRNA gene sequence analysis indicated that these strains were related to members of the genus Flavobacterium. Strain GH1-10(T) was most closely related to Flavobacterium psychrolimnae and Flavobacterium denitrificans, with sequence similarities of 95.9 and 95.2 %, respectively. Strain GH29-5(T) was most closely related to "Flavobacterium saliodium", F. denitrificans and Flavobacterium frigoris, with sequence similarities of 94.3, 92.5 and 92.5 %, respectively. The major cellular fatty acids of GH1-10(T) were iso-C(15 : 0), summed feature 3 (iso-C(15 : 0) 2-OH and/or C(16 : 1)omega7c) and iso-C(17 : 0) 3-OH, and those of GH29-5(T) were iso-C(15 : 0), iso-C(17 : 0) 3-OH, iso-C(15 : 1) G and iso-C(15 : 0) 3-OH. Both strains contained menaquinone with six isoprene units (MK-6) as the sole quinone. The DNA G+C contents of GH1-10(T) and GH29-5(T) were 35 and 39 mol%, respectively. Based on the phylogenetic and phenotypic data presented, it is concluded that the two bacteria represent two separate novel species of the genus Flavobacterium. The names proposed to accommodate these organisms are Flavobacterium daejeonense sp. nov., with type strain GH1-10(T) (=KACC 11422(T)=DSM 17708(T)), and Flavobacterium suncheonense sp. nov., with type strain GH29-5(T) (=KACC 11423(T)=DSM 17707(T)).

  5. Metatranscriptome Analysis of Aquifer Samples Reveals Unexpected Metabolic Lifestyles Relevant to Active Biogeochemical Cycling

    NASA Astrophysics Data System (ADS)

    Beller, H. R.; Jewell, T. N. M.; Karaoz, U.; Banfield, J. F.; Brodie, E.; Williams, K. H.

    2015-12-01

    Modern molecular ecology techniques are revealing the metabolic potential of uncultivated microorganisms, but there is still much to be learned about the actual biogeochemical roles of microbes that have cultivated relatives. Here, we present metatranscriptomic and metagenomic data from a field study that provides evidence of coupled redox processes that have not been documented in cultivated relatives and, indeed, represent strains with metabolic traits that are novel with respect to closely related isolates. The data come from omics analysis of groundwater samples collected during an experiment in which nitrate (a native electron acceptor) was injected into a perennially suboxic aquifer in Rifle (CO). Transcriptional data indicated that just two groups of chemolithoautotrophic bacteria accounted for a very large portion (~80%) of overall community gene expression: (1) members of the Fe(II)-oxidizing Gallionellaceae family and (2) strains of the S-oxidizing species, Sulfurimonas denitrificans. Metabolic lifestyles for Gallionellaceae strains that were novel compared to cultivated representatives included nitrate-dependent Fe(II) oxidation and S oxidation. Evidence for these metabolisms included highly correlated temporal expression in binned data of nitrate reductase (e.g., narGHI) genes (which have never been reported in Gallionellaceae genomes) and Fe(II) oxidation genes (e.g., mtoA) or S oxidation genes (e.g., dsrE, aprA). Of the two most active strains of S. denitrificans, only one showed strong expression of S oxidation genes, whereas the other was apparently using an unexpected (as-yet unidentified) primary electron donor. Transcriptional data added considerable interpretive value to this study, as (1) metagenomic data would not have highlighted these organisms, which had a disproportionately large role in community metabolism relative to their populations, and (2) co-expression of coupled pathway genes could not be predicted based solely on metagenomic data.

  6. Nitrate-Dependent Degradation of Acetone by Alicycliphilus and Paracoccus Strains and Comparison of Acetone Carboxylase Enzymes ▿

    PubMed Central

    Dullius, Carlos Henrique; Chen, Ching-Yuan; Schink, Bernhard

    2011-01-01

    A novel acetone-degrading, nitrate-reducing bacterium, strain KN Bun08, was isolated from an enrichment culture with butanone and nitrate as the sole sources of carbon and energy. The cells were motile short rods, 0.5 to 1 by 1 to 2 μm in size, which gave Gram-positive staining results in the exponential growth phase and Gram-negative staining results in the stationary-growth phase. Based on 16S rRNA gene sequence analysis, the isolate was assigned to the genus Alicycliphilus. Besides butanone and acetone, the strain used numerous fatty acids as substrates. An ATP-dependent acetone-carboxylating enzyme was enriched from cell extracts of this bacterium and of Alicycliphilus denitrificans K601T by two subsequent DEAE Sepharose column procedures. For comparison, acetone carboxylases were enriched from two additional nitrate-reducing bacterial species, Paracoccus denitrificans and P. pantotrophus. The products of the carboxylase reaction were acetoacetate and AMP rather than ADP. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of cell extracts and of the various enzyme preparations revealed bands corresponding to molecular masses of 85, 78, and 20 kDa, suggesting similarities to the acetone carboxylase enzymes described in detail for the aerobic bacterium Xanthobacter autotrophicus strain Py2 (85.3, 78.3, and 19.6 kDa) and the phototrophic bacterium Rhodobacter capsulatus. Protein bands were excised and compared by mass spectrometry with those of acetone carboxylases of aerobic bacteria. The results document the finding that the nitrate-reducing bacteria studied here use acetone-carboxylating enzymes similar to those of aerobic and phototrophic bacteria. PMID:21841031

  7. CCD Base Line Subtraction Algorithms

    SciTech Connect

    Kotov, I.V.; OConnor, P.; Kotov, A.; Frank, J.; Perevoztchikov, V.; Takacs, P.

    2010-06-28

    High statistics astronomical surveys require photometric accuracy on a few percent level. The accuracy of sensor calibration procedures should match this goal. The first step in calibration procedures is the base line subtraction. The accuracy and robustness of different base line subtraction techniques used for Charge Coupled Device (CCD) sensors are discussed.

  8. Plasma-based accelerator structures

    SciTech Connect

    Schroeder, Carl B.

    1999-12-01

    Plasma-based accelerators have the ability to sustain extremely large accelerating gradients, with possible high-energy physics applications. This dissertation further develops the theory of plasma-based accelerators by addressing three topics: the performance of a hollow plasma channel as an accelerating structure, the generation of ultrashort electron bunches, and the propagation of laser pulses is underdense plasmas.

  9. Competency Based Education: A Bibliography.

    ERIC Educational Resources Information Center

    Cox, Helen, Ed.

    This bibliography is a sampling of the types of publications which have appeared recently on the subject of competency based education. Emphasis is given to current (1970-76) materials and to adult career-related competencies. Titles specifically naming some phase of competency-based education are preferred above titles that emphasize performance,…

  10. Problem-Based Learning Tools

    ERIC Educational Resources Information Center

    Chin, Christine; Chia, Li-Gek

    2008-01-01

    One way of implementing project-based science (PBS) is to use problem-based learning (PBL), in which students formulate their own problems. These problems are often ill-structured, mirroring complex real-life problems where data are often messy and inclusive. In this article, the authors describe how they used PBL in a ninth-grade biology class in…

  11. Problem Based Learning in Science

    ERIC Educational Resources Information Center

    Pepper, Coral

    2009-01-01

    Problem based learning (PBL) is a recognised teaching and learning strategy used to engage students in deep rather than surface learning. It is also viewed as a successful strategy to align university courses with the real life professional work students are expected to undertake on graduation (Biggs, 2003). Problem based learning is practised…

  12. Agent-Based Literacy Theory

    ERIC Educational Resources Information Center

    McEneaney, John E.

    2006-01-01

    The purpose of this theoretical essay is to explore the limits of traditional conceptualizations of reader and text and to propose a more general theory based on the concept of a literacy agent. The proposed theoretical perspective subsumes concepts from traditional theory and aims to account for literacy online. The agent-based literacy theory…

  13. Theory-Based Stakeholder Evaluation

    ERIC Educational Resources Information Center

    Hansen, Morten Balle; Vedung, Evert

    2010-01-01

    This article introduces a new approach to program theory evaluation called theory-based stakeholder evaluation or the TSE model for short. Most theory-based approaches are program theory driven and some are stakeholder oriented as well. Practically, all of the latter fuse the program perceptions of the various stakeholder groups into one unitary…

  14. Reconstruction of Skull Base Defects.

    PubMed

    Klatt-Cromwell, Cristine N; Thorp, Brian D; Del Signore, Anthony G; Ebert, Charles S; Ewend, Matthew G; Zanation, Adam M

    2016-02-01

    "Endoscopic endonasal skull base surgery has dramatically changed and expanded over recent years due to significant advancements in instrumentation, techniques, and anatomic understanding. With these advances, the need for more robust skull base reconstructive techniques was vital. In this article, reconstructive options ranging from acellular grafts to vascular flaps are described, including the strengths, weaknesses, and common uses."

  15. Evidence-Based Language Practice

    ERIC Educational Resources Information Center

    Pollock, Eric J.

    2005-01-01

    The purpose of this paper was to examine evidence-based procedures in medicine and to demonstrate that the same protocols can be used in English language instruction. In the evidence-based methodology, studies are divided into those that address specific language problems. Integrated studies are presented as a systematic overview, meta-analysis,…

  16. Cloud-Based Data Storage

    ERIC Educational Resources Information Center

    Waters, John K.

    2011-01-01

    The vulnerability and inefficiency of backing up data on-site is prompting school districts to switch to more secure, less troublesome cloud-based options. District auditors are pushing for a better way to back up their data than the on-site, tape-based system that had been used for years. About three years ago, Hendrick School District in…

  17. Discipline Based Arts Education. [Videotape].

    ERIC Educational Resources Information Center

    Phillips, Lori

    This 5-part video lesson deals with discipline-based art education in the elementary school. The video features a university professor who is a specialist in methods and the integration of art into the elementary classroom; each part of the video lesson is 30 minutes in length. First defining discipline-based art education as an approach, not a…

  18. Radiological Worker Computer Based Training

    2003-02-06

    Argonne National Laboratory has developed an interactive computer based training (CBT) version of the standardized DOE Radiological Worker training program. This CD-ROM based program utilizes graphics, animation, photographs, sound and video to train users in ten topical areas: radiological fundamentals, biological effects, dose limits, ALARA, personnel monitoring, controls and postings, emergency response, contamination controls, high radiation areas, and lessons learned.

  19. Using dBase III.

    ERIC Educational Resources Information Center

    Evans, Janet; And Others

    1986-01-01

    Four articles on dBASE III include three on library applications: a photocopy invoicing system for interlibrary loan, a vertical file subject headings list program, and a subject index to statistical resources. Another article explains the differences between interpreters and compilers and the advantages of the Clipper compiler for dBASE III. (EM)

  20. Lignin-Based Thermoplastic Materials.

    PubMed

    Wang, Chao; Kelley, Stephen S; Venditti, Richard A

    2016-04-21

    Lignin-based thermoplastic materials have attracted increasing interest as sustainable, cost-effective, and biodegradable alternatives for petroleum-based thermoplastics. As an amorphous thermoplastic material, lignin has a relatively high glass-transition temperature and also undergoes radical-induced self-condensation at high temperatures, which limits its thermal processability. Additionally, lignin-based materials are usually brittle and exhibit poor mechanical properties. To improve the thermoplasticity and mechanical properties of technical lignin, polymers or plasticizers are usually integrated with lignin by blending or chemical modification. This Review attempts to cover the reported approaches towards the development of lignin-based thermoplastic materials on the basis of published information. Approaches reviewed include plasticization, blending with miscible polymers, and chemical modifications by esterification, etherification, polymer grafting, and copolymerization. Those lignin-based thermoplastic materials are expected to show applications as engineering plastics, polymeric foams, thermoplastic elastomers, and carbon-fiber precursors. PMID:27059111

  1. Fundamental base closure environmental principles

    SciTech Connect

    Yim, R.A.

    1994-12-31

    Military base closures present a paradox. The rate, scale and timing of military base closures is historically unique. However, each base itself typically does not present unique problems. Thus, the challenge is to design innovative solutions to base redevelopment and remediation issues, while simultaneously adopting common, streamlined or pre-approved strategies to shared problems. The author presents six environmental principles that are fundamental to base closure. They are: remediation not clean up; remediation will impact reuse; reuse will impact remediation; remediation and reuse must be coordinated; environmental contamination must be evaluated as any other initial physical constraint on development, not as an overlay after plans are created; and remediation will impact development, financing and marketability.

  2. Lignin-Based Thermoplastic Materials.

    PubMed

    Wang, Chao; Kelley, Stephen S; Venditti, Richard A

    2016-04-21

    Lignin-based thermoplastic materials have attracted increasing interest as sustainable, cost-effective, and biodegradable alternatives for petroleum-based thermoplastics. As an amorphous thermoplastic material, lignin has a relatively high glass-transition temperature and also undergoes radical-induced self-condensation at high temperatures, which limits its thermal processability. Additionally, lignin-based materials are usually brittle and exhibit poor mechanical properties. To improve the thermoplasticity and mechanical properties of technical lignin, polymers or plasticizers are usually integrated with lignin by blending or chemical modification. This Review attempts to cover the reported approaches towards the development of lignin-based thermoplastic materials on the basis of published information. Approaches reviewed include plasticization, blending with miscible polymers, and chemical modifications by esterification, etherification, polymer grafting, and copolymerization. Those lignin-based thermoplastic materials are expected to show applications as engineering plastics, polymeric foams, thermoplastic elastomers, and carbon-fiber precursors.

  3. Object-based media and stream-based computing

    NASA Astrophysics Data System (ADS)

    Bove, V. Michael, Jr.

    1998-03-01

    Object-based media refers to the representation of audiovisual information as a collection of objects - the result of scene-analysis algorithms - and a script describing how they are to be rendered for display. Such multimedia presentations can adapt to viewing circumstances as well as to viewer preferences and behavior, and can provide a richer link between content creator and consumer. With faster networks and processors, such ideas become applicable to live interpersonal communications as well, creating a more natural and productive alternative to traditional videoconferencing. In this paper is outlined an example of object-based media algorithms and applications developed by my group, and present new hardware architectures and software methods that we have developed to enable meeting the computational requirements of object- based and other advanced media representations. In particular we describe stream-based processing, which enables automatic run-time parallelization of multidimensional signal processing tasks even given heterogenous computational resources.

  4. Value-based metrics and Internet-based enterprises

    NASA Astrophysics Data System (ADS)

    Gupta, Krishan M.

    2001-10-01

    Within the last few years, a host of value-based metrics like EVA, MVA, TBR, CFORI, and TSR have evolved. This paper attempts to analyze the validity and applicability of EVA and Balanced Scorecard for Internet based organizations. Despite the collapse of the dot-com model, the firms engaged in e- commerce continue to struggle to find new ways to account for customer-base, technology, employees, knowledge, etc, as part of the value of the firm. While some metrics, like the Balance Scorecard are geared towards internal use, others like EVA are for external use. Value-based metrics are used for performing internal audits as well as comparing firms against one another; and can also be effectively utilized by individuals outside the firm looking to determine if the firm is creating value for its stakeholders.

  5. Coronavirus phylogeny based on triplets of nucleic acids bases

    NASA Astrophysics Data System (ADS)

    Liao, Bo; Liu, Yanshu; Li, Renfa; Zhu, Wen

    2006-04-01

    We considered the fully overlapping triplets of nucleotide bases and proposed a 2D graphical representation of protein sequences consisting of 20 amino acids and a stop code. Based on this 2D graphical representation, we outlined a new approach to analyze the phylogenetic relationships of coronaviruses by constructing a covariance matrix. The evolutionary distances are obtained through measuring the differences among the two-dimensional curves.

  6. Execution-Based Model Checking of Interrupt-Based Systems

    NASA Technical Reports Server (NTRS)

    Drusinsky, Doron; Havelund, Klaus

    2003-01-01

    Execution-based model checking (EMC) is a verification technique based on executing a multi-threaded/multiprocess program repeatedly in a systematic manner in order to explore the different interleavings of the program. This is in contrast to traditional model checking, where a model of a system is analyzed Several execution-based model-checking tools exist at this point, such as for example Verisoft and Java PathFinder. The most common formal specification languages used by EMC tools are un- timed, either just assertions, or linear-time temporal logic (LTL). An alternative verification technique is Runtime Execution Monitoring (REM), which is based on monitor- ing the execution of a program, checking that the execution trace conforms to a requirement specification. The Temporal Rover and DBRover are such tools. They provide a very rich specification language, being an extension of LTL with real-time constraints and time-series. We show how execution-based model checking, combined with runtime execution monitoring, can be used for the verification of a large class of safety critical systems commonly known as interrupt-based systems. The proposed approach is novel in that: (i) it supports model checking of a large class of applications not practically verifiable using conventional EMC tools, (ii) it supports verification of LTL assertions extended with real-time and time-series constraints, and (iii) it supports the verification of custom schedulers.

  7. Mineral-resource data bases

    USGS Publications Warehouse

    ,

    1997-01-01

    Data bases are essential for modern scientific research. The new and exciting work being done in the Mineral Resource Program in the U.S. Geological Survey (USGS) usually begins with the question, "Where are the known deposits?" A mineral-resource data base containing this type of information and more can be useful not just to USGS scientists, but to anyone who needs such data. Users of the data bases from outside the USGS include mining and exploration companies, environmental groups, academia, other Federal Agencies, and the general public. At present, the USGS has two large mineral-resource data bases, MRDS (Mineral Resource Data System) and MAS (Minerals Availability System). MRDS was built and is mamtained by the USGS, and MAS was built and maintained by the Bureau of Mines. In 1996, after the Bureau was abolished, MAS was transferred to the USGS. The two data bases were compiled for different purposes and contain very different mformation. For instance, MAS contains information on costs, details of mining methods, and feasibility studies. MRDS has mineralogical and geologic data that are not contained in MAS. Because they are both mineral-resource data bases, however, they contain some information in common, such as location, name(s) of sites, and commodities present. Both data bases are international in scope, and both are quite large. MRDS contains over 110,000 records, while MAS has over 220,000. One reason that MAS has more records is that it contains information on smelters, mill sites, and fossil fuel sites, as well as mineral- resource sites. The USGS is working to combine the information in both data bases. This is a large undertaking that will require some years to complete. In the interim, information from both data bases will still be available

  8. DSP based data acquisition system

    NASA Astrophysics Data System (ADS)

    Prasad, Peeyush; Subrahmanya, C. R.

    A new DSP based data acquisition system based on commercial, off-the-shelf available hardware is presented. The board features friendly high speed input and output interfaces, a powerful DSP and hardware offloading of essing to an FPGA. For flexibility of development, a soft realtime linux based kernel popular among embedded developers (uClinux) has been ported on the board.The development environment is fully open source both for the target as well as for host software. The motivation behind creating the system, the development ess and the current status is described. Some results obtained with this system as used in typical applications are also presented.

  9. Nanocrystal-based Optoelectronic Devices

    NASA Astrophysics Data System (ADS)

    Evans, Kenneth; Herzog, Joseph; Ward, Daniel; Natelson, Douglas

    2012-02-01

    Optoelectronic devices capable of detecting and emitting light on a scale well below its wavelength could have a profound impact on basic and applied experimental research in light-based electronics, on-demand photon generation, and for studying poorly understood quantum phenomena such as blinking and spectral wandering. We present a fabrication procedure for ultrasmall, nanocrystal optoelectronic devices based on self-assembled layers of quantum dots in plasmonically-active gold nanogaps. We provide preliminary experimental results which examine the possibility for surfaced-enhanced fluorescence, subwavelength detection and emission of light as well as plasmon-based optical trapping in these systems.

  10. New carbohydrate-based materials

    SciTech Connect

    Callstrom, M.R.

    1992-07-01

    We have prepared a series of new carbohydrate-based materials based on the use of carbohydrates as a template for the introduction of functionality to polymeric materials with complete regio- and stereochemical control. The synthesis of these new materials by the use of chemical and enzymatic methods allows for the rational design of new materials based on the properties of the monomeric subunit. These materials have potential applications that range from their use in enhanced oil recovery to biodegradable plastics to biological applications including targeted drug delivery and enzyme stabilization.

  11. Chip-based droplet sorting

    DOEpatents

    Beer, Neil Reginald; Lee, Abraham; Hatch, Andrew

    2014-07-01

    A non-contact system for sorting monodisperse water-in-oil emulsion droplets in a microfluidic device based on the droplet's contents and their interaction with an applied electromagnetic field or by identification and sorting.

  12. Point-based manifold harmonics.

    PubMed

    Liu, Yang; Prabhakaran, Balakrishnan; Guo, Xiaohu

    2012-10-01

    This paper proposes an algorithm to build a set of orthogonal Point-Based Manifold Harmonic Bases (PB-MHB) for spectral analysis over point-sampled manifold surfaces. To ensure that PB-MHB are orthogonal to each other, it is necessary to have symmetrizable discrete Laplace-Beltrami Operator (LBO) over the surfaces. Existing converging discrete LBO for point clouds, as proposed by Belkin et al., is not guaranteed to be symmetrizable. We build a new point-wisely discrete LBO over the point-sampled surface that is guaranteed to be symmetrizable, and prove its convergence. By solving the eigen problem related to the new operator, we define a set of orthogonal bases over the point cloud. Experiments show that the new operator is converging better than other symmetrizable discrete Laplacian operators (such as graph Laplacian) defined on point-sampled surfaces, and can provide orthogonal bases for further spectral geometric analysis and processing tasks.

  13. Pitch based foam with particulate

    DOEpatents

    Klett, James W.

    2001-01-01

    A thermally conductive, pitch based foam composite having a particulate content. The particulate alters the mechanical characteristics of the foam without severely degrading the foam thermal conductivity. The composite is formed by mixing the particulate with pitch prior to foaming.

  14. Mechanism of base-catalyzed Schiff base deprotonation in halorhodopsin

    SciTech Connect

    Lanyi, J.K.

    1986-10-21

    It has been shown earlier that the deprotonation of the Schiff base of illuminated halorhodopsin proceeds with a much lower pK/sub a/ than that of the unilluminated pigment and the reversible protonation change is catalyzed by azide and cyanate. The authors have studied the kinetics of the proton-transfer events with flash spectroscopy and compared a variety of anionic bases with different pK/sub a/ with regard to the apparent binding constants and the catalytic activities. The results suggest a general base catalysis mechanism in which the anionic bases bind with apparently low affinity to halorhodopsin, although with some degree of size- and/or shape-dependent specificity. The locus of the catalysis is accessible from the cytoplasmic side of the membrane and is not at site I, where various anions bind and shift the pK/sub a/ of the deprotonation. Neither is it at site II, where a few specific anions (like chloride) bind to the all-trans pigment. It may be concluded that while the all-trans pigment loses its Schiff base proton very rapidly at its pK/sub a/, there is a kinetic barrier to this deprotonation in the 13-cis photointermediate that can be partially overcome by the reversible protonation of an extrinsic anionic base, which shuttles protons between the interior of the protein and the aqueous medium. The need for an extrinsic proton acceptor for efficient deprotonation of the Schiff base of halorhodopsin is one of the main differences between this pigment and the analogous retinal protein, bacteriorhodopsin.

  15. Imaging of skull base tumours.

    PubMed

    Thust, Stefanie Catherine; Yousry, Tarek

    2016-01-01

    The skull base is a highly complex and difficult to access anatomical region, which constitutes a relatively common site for neoplasms. Imaging plays a central role in establishing the differential diagnosis, to determine the anatomic tumour spread and for operative planning. All skull base imaging should be performed using thin-section multiplanar imaging, whereby CT and MRI can be considered complimentary. An interdisciplinary team approach is central to improve the outcome of these challenging tumours.

  16. Osmium-Based Pyrimidine Contrast Tags for Enhanced Nanopore-Based DNA Base Discrimination

    PubMed Central

    Henley, Robert Y.; Vazquez-Pagan, Ana G.; Johnson, Michael; Kanavarioti, Anastassia; Wanunu, Meni

    2015-01-01

    Nanopores are a promising platform in next generation DNA sequencing. In this platform, an individual DNA strand is threaded into nanopore using an electric field, and enzyme-based ratcheting is used to move the strand through the detector. During this process the residual ion current through the pore is measured, which exhibits unique levels for different base combinations inside the pore. While this approach has shown great promise, accuracy is not optimal because the four bases are chemically comparable to one another, leading to small differences in current obstruction. Nucleobase-specific chemical tagging can be a viable approach to enhancing the contrast between different bases in the sequence. Herein we show that covalent modification of one or both of the pyrimidine bases by an osmium bipyridine complex leads to measureable differences in the blockade amplitudes of DNA molecules. We qualitatively determine the degree of osmylation of a DNA strand by passing it through a solid-state nanopore, and are thus able to gauge T and C base content. In addition, we show that osmium bipyridine reacts with dsDNA, leading to substantially different current blockade levels than exhibited for bare dsDNA. This work serves as a proof of principle for nanopore sequencing and mapping via base-specific DNA osmylation. PMID:26657869

  17. Osmium-Based Pyrimidine Contrast Tags for Enhanced Nanopore-Based DNA Base Discrimination.

    PubMed

    Henley, Robert Y; Vazquez-Pagan, Ana G; Johnson, Michael; Kanavarioti, Anastassia; Wanunu, Meni

    2015-01-01

    Nanopores are a promising platform in next generation DNA sequencing. In this platform, an individual DNA strand is threaded into nanopore using an electric field, and enzyme-based ratcheting is used to move the strand through the detector. During this process the residual ion current through the pore is measured, which exhibits unique levels for different base combinations inside the pore. While this approach has shown great promise, accuracy is not optimal because the four bases are chemically comparable to one another, leading to small differences in current obstruction. Nucleobase-specific chemical tagging can be a viable approach to enhancing the contrast between different bases in the sequence. Herein we show that covalent modification of one or both of the pyrimidine bases by an osmium bipyridine complex leads to measureable differences in the blockade amplitudes of DNA molecules. We qualitatively determine the degree of osmylation of a DNA strand by passing it through a solid-state nanopore, and are thus able to gauge T and C base content. In addition, we show that osmium bipyridine reacts with dsDNA, leading to substantially different current blockade levels than exhibited for bare dsDNA. This work serves as a proof of principle for nanopore sequencing and mapping via base-specific DNA osmylation.

  18. Osmium-Based Pyrimidine Contrast Tags for Enhanced Nanopore-Based DNA Base Discrimination.

    PubMed

    Henley, Robert Y; Vazquez-Pagan, Ana G; Johnson, Michael; Kanavarioti, Anastassia; Wanunu, Meni

    2015-01-01

    Nanopores are a promising platform in next generation DNA sequencing. In this platform, an individual DNA strand is threaded into nanopore using an electric field, and enzyme-based ratcheting is used to move the strand through the detector. During this process the residual ion current through the pore is measured, which exhibits unique levels for different base combinations inside the pore. While this approach has shown great promise, accuracy is not optimal because the four bases are chemically comparable to one another, leading to small differences in current obstruction. Nucleobase-specific chemical tagging can be a viable approach to enhancing the contrast between different bases in the sequence. Herein we show that covalent modification of one or both of the pyrimidine bases by an osmium bipyridine complex leads to measureable differences in the blockade amplitudes of DNA molecules. We qualitatively determine the degree of osmylation of a DNA strand by passing it through a solid-state nanopore, and are thus able to gauge T and C base content. In addition, we show that osmium bipyridine reacts with dsDNA, leading to substantially different current blockade levels than exhibited for bare dsDNA. This work serves as a proof of principle for nanopore sequencing and mapping via base-specific DNA osmylation. PMID:26657869

  19. Rule-based simulation models

    NASA Technical Reports Server (NTRS)

    Nieten, Joseph L.; Seraphine, Kathleen M.

    1991-01-01

    Procedural modeling systems, rule based modeling systems, and a method for converting a procedural model to a rule based model are described. Simulation models are used to represent real time engineering systems. A real time system can be represented by a set of equations or functions connected so that they perform in the same manner as the actual system. Most modeling system languages are based on FORTRAN or some other procedural language. Therefore, they must be enhanced with a reaction capability. Rule based systems are reactive by definition. Once the engineering system has been decomposed into a set of calculations using only basic algebraic unary operations, a knowledge network of calculations and functions can be constructed. The knowledge network required by a rule based system can be generated by a knowledge acquisition tool or a source level compiler. The compiler would take an existing model source file, a syntax template, and a symbol table and generate the knowledge network. Thus, existing procedural models can be translated and executed by a rule based system. Neural models can be provide the high capacity data manipulation required by the most complex real time models.

  20. NLS base heating CFD analysis

    NASA Technical Reports Server (NTRS)

    Ascoli, Edward P.; Heiba, Adel H.; Hsu, Yann-Fu; Lagnado, Ronald R.; Lynch, Edward D.

    1993-01-01

    Concerns raised over possible base heating effects on the National Launch System (NLS) 1.5 stage reference vehicle resulted in the use of CFD as a predictive analysis tool. The objective established was to obtain good engineering solutions to describe the base region flowfields at 10,000 ft and 50,000 ft altitudes. The Rockwell USA CFD code was employed with a zero-equation turbulence model and a four species, 1 step chemical kinetics package. Three solutions were generated for the specified altitudes on coarse and fine grids. CFD results show the base region flowfields to be highly three dimensional in character. At the 10,000 ft altitude, plumes contract soon after exiting the nozzles and do not interact with each other. No mechanism was identified for driving hot gas back into the base region and no significant amounts of hydrogen or water were found in the base region. Consequently, surface temperatures were all near the ambient level. At 50,000 ft, the nozzle exhaust plumes begin to interact, particularly those of the two inboard engines which are closer together. A small amount of hot gas is recirculated between the inboard nozzles near the nozzle exit plane. As a result, base region surface temperatures are slightly elevated, but still remain well within the design guideline of 1000 R.

  1. Current NASA lunar base concepts

    NASA Technical Reports Server (NTRS)

    Roberts, Barney B.; Connolly, John F.

    1990-01-01

    The NASA Office of Aeronautics, Exploration, and Technology has completed a Systems Engineering and Integration effort to define a point design for an evolving lunar base that supports substantial science, exploration, and resource production objectives. This study addressed the following: systems level design; element requirements and conceptual design; assessments of precursor and technology needs; and operations concepts. The central base is assumed to be located equatorially on the lunar nearside north of the crater Moltke in Mare Tranquilliatis. The study considers an aggressive case with three main phases. The initial Man-Tended Phase established basic enabling facilities that include a modular habitat that periodically houses a crew of four. During the Experimental Phase, the base becomes permanently manned with the construction of a larger habitat that provides augmented workshop and laboratory volumes and housing for crew. The Operational Phase expands base capabilities to a substantially mature level while reducing reliance on Earth. The analysis classifies base characteristics into several major functional areas: Human Systems; Assembly and Construction; Energy Management; Launch and Landing; Surface Transportation; In-Situ Resources Utilization; User Accommodations; and Telecommunications, Navigation, and Information Management. Results of various NASA-sponsored studies were synthesized to meet requirements. The system level architecture was determined, the physical layout was developed from a set of proximity criteria and related functions, and the evlotuionary path of the base was analyzed. Conclusions include a summary of technology needs, design drivers, high leverage items, and important issues.

  2. Home-based versus centre-based cardiac rehabilitation

    PubMed Central

    Taylor, Rod S; Dalal, Hayes; Jolly, Kate; Moxham, Tiffany; Zawada, Anna

    2014-01-01

    Background The burden of cardiovascular disease world-wide is one of great concern to patients and health care agencies alike. Traditionally centre-based cardiac rehabilitation (CR) programmes are offered to individuals after cardiac events to aid recovery and prevent further cardiac illness. Home-based cardiac rehabilitation programmes have been introduced in an attempt to widen access and participation. Objectives To determine the effectiveness of home-based cardiac rehabilitation programmes compared with supervised centre-based cardiac rehabilitation on mortality and morbidity, health-related quality of life and modifiable cardiac risk factors in patients with coronary heart disease. Search methods We updated the search of a previous review by searching the Cochrane Central Register of Controlled Trials (CENTRAL) in The Cochrane Library (2007, Issue 4), MEDLINE, EMBASE and CINAHL from 2001 to January 2008. We checked reference lists and sought advice from experts. No language restrictions were applied. Selection criteria Randomised controlled trials (RCTs) that compared centre-based cardiac rehabilitation (e.g. hospital, gymnasium, sports centre) with home-based programmes, in adults with myocardial infarction, angina, heart failure or who had undergone revascularisation. Data collection and analysis Studies were selected independently by two reviewers, and data extracted by a single reviewer and checked by a second one. Authors were contacted where possible to obtain missing information. Main results Twelve studies (1,938 participants) met the inclusion criteria. The majority of studies recruited a lower risk patient following an acute myocardial infarction (MI) and revascularisation. There was no difference in outcomes of home- versus centre-based cardiac rehabilitation in mortality risk ratio (RR) was 1.31 (95% confidence interval (C) 0.65 to 2.66), cardiac events, exercise capacity standardised mean difference (SMD) −0.11 (95% CI −0.35 to 0.13), as well

  3. Case-Based Tutoring from a Medical Knowledge Base

    PubMed Central

    Chin, Homer L.

    1988-01-01

    The past decade has seen the emergence of programs that make use of large knowledge bases to assist physicians in diagnosis within the general field of internal medicine. One such program, Internist-I, contains knowledge about over 600 diseases, covering a significant proportion of internal medicine. This paper describes the process of converting a subset of this knowledge base--in the area of cardiovascular diseases--into a probabilistic format, and the use of this resulting knowledge base to teach medical diagnostic knowledge. The system (called KBSimulator--for Knowledge-Based patient Simulator) generates simulated patient cases and uses these cases as a focal point from which to teach medical knowledge. It interacts with the student in a mixed-initiative fashion, presenting patients for the student to diagnose, and allowing the student to obtain further information on his/her own initiative in the context of that patient case. The system scores the student, and uses these scores to form a rudimentary model of the student. This resulting model of the student is then used to direct the generation of subsequent patient cases. This project demonstrates the feasibility of building an intelligent, flexible instructional system that uses a knowledge base constructed primarily for medical diagnosis.

  4. Image denoising filter based on patch-based difference refinement

    NASA Astrophysics Data System (ADS)

    Park, Sang Wook; Kang, Moon Gi

    2012-06-01

    In the denoising literature, research based on the nonlocal means (NLM) filter has been done and there have been many variations and improvements regarding weight function and parameter optimization. Here, a NLM filter with patch-based difference (PBD) refinement is presented. PBD refinement, which is the weighted average of the PBD values, is performed with respect to the difference images of all the locations in a refinement kernel. With refined and denoised PBD values, pattern adaptive smoothing threshold and noise suppressed NLM filter weights are calculated. Owing to the refinement of the PBD values, the patterns are divided into flat regions and texture regions by comparing the sorted values in the PBD domain to the threshold value including the noise standard deviation. Then, two different smoothing thresholds are utilized for each region denoising, respectively, and the NLM filter is applied finally. Experimental results of the proposed scheme are shown in comparison with several state-of-the-arts NLM based denoising methods.

  5. [Laser-based radiometric calibration].

    PubMed

    Li, Zhi-gang; Zheng, Yu-quan

    2014-12-01

    Increasingly higher demands are put forward to spectral radiometric calibration accuracy and the development of new tunable laser based spectral radiometric calibration technology is promoted, along with the development of studies of terrestrial remote sensing, aeronautical and astronautical remote sensing, plasma physics, quantitative spectroscopy, etc. Internationally a number of national metrology scientific research institutes have built tunable laser based spectral radiometric calibration facilities in succession, which are traceable to cryogenic radiometers and have low uncertainties for spectral responsivity calibration and characterization of detectors and remote sensing instruments in the UK, the USA, Germany, etc. Among them, the facility for spectral irradiance and radiance responsivity calibrations using uniform sources (SIRCCUS) at the National Institute of Standards and Technology (NIST) in the USA and the Tunable Lasers in Photometry (TULIP) facility at the Physikalisch-Technische Bundesanstalt (PTB) in Germany have more representatives. Compared with lamp-monochromator systems, laser based spectral radiometric calibrations have many advantages, such as narrow spectral bandwidth, high wavelength accuracy, low calibration uncertainty and so on for radiometric calibration applications. In this paper, the development of laser-based spectral radiometric calibration and structures and performances of laser-based radiometric calibration facilities represented by the National Physical Laboratory (NPL) in the UK, NIST and PTB are presented, technical advantages of laser-based spectral radiometric calibration are analyzed, and applications of this technology are further discussed. Laser-based spectral radiometric calibration facilities can be widely used in important system-level radiometric calibration measurements with high accuracy, including radiance temperature, radiance and irradiance calibrations for space remote sensing instruments, and promote the

  6. [Laser-based radiometric calibration].

    PubMed

    Li, Zhi-gang; Zheng, Yu-quan

    2014-12-01

    Increasingly higher demands are put forward to spectral radiometric calibration accuracy and the development of new tunable laser based spectral radiometric calibration technology is promoted, along with the development of studies of terrestrial remote sensing, aeronautical and astronautical remote sensing, plasma physics, quantitative spectroscopy, etc. Internationally a number of national metrology scientific research institutes have built tunable laser based spectral radiometric calibration facilities in succession, which are traceable to cryogenic radiometers and have low uncertainties for spectral responsivity calibration and characterization of detectors and remote sensing instruments in the UK, the USA, Germany, etc. Among them, the facility for spectral irradiance and radiance responsivity calibrations using uniform sources (SIRCCUS) at the National Institute of Standards and Technology (NIST) in the USA and the Tunable Lasers in Photometry (TULIP) facility at the Physikalisch-Technische Bundesanstalt (PTB) in Germany have more representatives. Compared with lamp-monochromator systems, laser based spectral radiometric calibrations have many advantages, such as narrow spectral bandwidth, high wavelength accuracy, low calibration uncertainty and so on for radiometric calibration applications. In this paper, the development of laser-based spectral radiometric calibration and structures and performances of laser-based radiometric calibration facilities represented by the National Physical Laboratory (NPL) in the UK, NIST and PTB are presented, technical advantages of laser-based spectral radiometric calibration are analyzed, and applications of this technology are further discussed. Laser-based spectral radiometric calibration facilities can be widely used in important system-level radiometric calibration measurements with high accuracy, including radiance temperature, radiance and irradiance calibrations for space remote sensing instruments, and promote the

  7. Antibacterial and antifungal metal based triazole Schiff bases.

    PubMed

    Chohan, Zahid H; Hanif, Muhammad

    2013-10-01

    A new series of four biologically active triazole derived Schiff base ligands (L(1)-L(4)) and their cobalt(II), nickel(II), copper(II) and zinc(II) complexes (1-16) have been synthesized and characterized. The ligands were prepared by the condensation reaction of 3-amino-5-methylthio-1H-1,2,4-triazole with chloro-, bromo- and nitro-substituted 2-hydroxybenzaldehyde in an equimolar ratio. The antibacterial and antifungal bioactivity data showed the metal(II) complexes to be more potent antibacterial and antifungal than the parent Schiff bases against one or more bacterial and fungal species.

  8. Summary of LOGDEX data base

    SciTech Connect

    Hill, T.; Sepehrnoori, K.

    1981-08-01

    A summary of LOGDEX, the digitized well log data base maintained by the Center for Energy Studies at The University of Texas at Austin is presented. These well logs were obtained from various oil companies and then converted from paper well logs to numeric information on magnetic computer tapes for input into the well log data base. This data base serves as a resource for application programs in the study of geopressured geothermal energy resources, for well logging research, and for geological research. Currently the location and scope of well log data that may be found within the LOGDEX data base are limited to wells along the Texas-Louisiana Gulf Coast that are known to have a potential as a geopressured geothermal energy resource. Additionally the location of these wells in that area is highly localized into areas that have been defined by Department of Energy researchers as having a high potential for geopressured geothermal energy. The LOGDEX data base currently contains data from more than 350 wells, representing more than 1600 logs and 16,600,000 curve feet of data. For quick reference to a given log, the summary listing has been indexed into seven divisions: well classification, location by county or parish, curve type, log type, operators, location by state, and well names. These indexes are arranged alphabetically and cross-referenced by page number.

  9. Automated knowledge-base refinement

    NASA Technical Reports Server (NTRS)

    Mooney, Raymond J.

    1994-01-01

    Over the last several years, we have developed several systems for automatically refining incomplete and incorrect knowledge bases. These systems are given an imperfect rule base and a set of training examples and minimally modify the knowledge base to make it consistent with the examples. One of our most recent systems, FORTE, revises first-order Horn-clause knowledge bases. This system can be viewed as automatically debugging Prolog programs based on examples of correct and incorrect I/O pairs. In fact, we have already used the system to debug simple Prolog programs written by students in a programming language course. FORTE has also been used to automatically induce and revise qualitative models of several continuous dynamic devices from qualitative behavior traces. For example, it has been used to induce and revise a qualitative model of a portion of the Reaction Control System (RCS) of the NASA Space Shuttle. By fitting a correct model of this portion of the RCS to simulated qualitative data from a faulty system, FORTE was also able to correctly diagnose simple faults in this system.

  10. Imaging of skull base lesions.

    PubMed

    Kelly, Hillary R; Curtin, Hugh D

    2016-01-01

    Skull base imaging requires a thorough knowledge of the complex anatomy of this region, including the numerous fissures and foramina and the major neurovascular structures that traverse them. Computed tomography (CT) and magnetic resonance imaging (MRI) play complementary roles in imaging of the skull base. MR is the preferred modality for evaluation of the soft tissues, the cranial nerves, and the medullary spaces of bone, while CT is preferred for demonstrating thin cortical bone structure. The anatomic location and origin of a lesion as well as the specific CT and MR findings can often narrow the differential diagnosis to a short list of possibilities. However, the primary role of the imaging specialist in evaluating the skull base is usually to define the extent of the lesion and determine its relationship to vital neurovascular structures. Technologic advances in imaging and radiation therapy, as well as surgical technique, have allowed for more aggressive approaches and improved outcomes, further emphasizing the importance of precise preoperative mapping of skull base lesions via imaging. Tumors arising from and affecting the cranial nerves at the skull base are considered here. PMID:27432686

  11. Imaging of skull base lesions.

    PubMed

    Kelly, Hillary R; Curtin, Hugh D

    2016-01-01

    Skull base imaging requires a thorough knowledge of the complex anatomy of this region, including the numerous fissures and foramina and the major neurovascular structures that traverse them. Computed tomography (CT) and magnetic resonance imaging (MRI) play complementary roles in imaging of the skull base. MR is the preferred modality for evaluation of the soft tissues, the cranial nerves, and the medullary spaces of bone, while CT is preferred for demonstrating thin cortical bone structure. The anatomic location and origin of a lesion as well as the specific CT and MR findings can often narrow the differential diagnosis to a short list of possibilities. However, the primary role of the imaging specialist in evaluating the skull base is usually to define the extent of the lesion and determine its relationship to vital neurovascular structures. Technologic advances in imaging and radiation therapy, as well as surgical technique, have allowed for more aggressive approaches and improved outcomes, further emphasizing the importance of precise preoperative mapping of skull base lesions via imaging. Tumors arising from and affecting the cranial nerves at the skull base are considered here.

  12. Photography-based image generator

    NASA Astrophysics Data System (ADS)

    Dalton, Nicholas M.; Deering, Charles S.

    1989-09-01

    A two-channel Photography Based Image Generator system was developed to drive the Helmet Mounted Laser Projector at the Naval Training System Center at Orlando, Florida. This projector is a two-channel system that displays a wide field-of-view color image with a high-resolution inset to efficiently match the pilot's visual capability. The image generator is a derivative of the LTV-developed visual system installed in the A-7E Weapon System Trainer at NAS Cecil Field. The Photography Based Image Generator is based on patented LTV technology for high resolution, multi-channel, real world visual simulation. Special provisions were developed for driving the NTSC-developed and patented Helmet Mounted Laser Projector. These include a special 1023-line raster format, an electronic image blending technique, spherical lens mapping for dome projection, a special computer interface for head/eye tracking and flight parameters, special software, and a number of data bases. Good gaze angle tracking is critical to the use of the NTSC projector in a flight simulation environment. The Photography Based Image Generator provides superior dynamic response by performing a relatively simple perspective transformation on stored, high-detail photography instead of generating this detail by "brute force" computer image generation methods. With this approach, high detail can be displayed and updated at the television field rate (60 Hz).

  13. An Examination of the Bases of Evidence-Based Interventions.

    ERIC Educational Resources Information Center

    Wampold, Bruce E.

    2002-01-01

    School psychology has proposed a system to aid in the identification of evidence-based interventions (Kratochwill & Stoiber, this issue; Lewis-Snyder, Stoiber, & Kratochwill, this issue; Shernoff, Kratochwill, & Stoiber, this issue). In this commentary, issues related to the politics of exclusion, design and theory, methods, and multiculturalism…

  14. Secure Base Priming Diminishes Conflict-Based Anger and Anxiety

    PubMed Central

    Koren, Tamara; Bartholomew, Kim

    2016-01-01

    This study examines the impact of a visual representation of a secure base (i.e. a secure base prime) on attenuating experimentally produced anger and anxiety. Specifically, we examined the assuaging of negative emotions through exposure to an image of a mother-infant embrace or a heterosexual couple embracing. Subjects seated at a computer terminal rated their affect (Pre Affect) using the Affect Adjective Checklist (AAC) then listened to two sets of intense two person conflicts. After the first conflict exposure they rated affect again (Post 1 AAC). Following the second exposure they saw a blank screen (control condition), pictures of everyday objects (distraction condition) or a photo of two people embracing (Secure Base Prime condition). They then reported emotions using the Post 2 AAC. Compared to either control or distraction subjects, Secure Base Prime (SBP) subjects reported significantly less anger and anxiety. These results were then replicated using an internet sample with control, SBP and two new controls: Smiling Man (to control for expression of positive affect) and Cold Mother (an unsmiling mother with infant). The SBP amelioration of anger and anxiety was replicated with the internet sample. No control groups produced this effect, which was generated only by a combination of positive affect in a physically embracing dyad. The results are discussed in terms of attachment theory and research on spreading activation. PMID:27606897

  15. Secure Base Priming Diminishes Conflict-Based Anger and Anxiety.

    PubMed

    Dutton, Donald G; Lane, René A; Koren, Tamara; Bartholomew, Kim

    2016-01-01

    This study examines the impact of a visual representation of a secure base (i.e. a secure base prime) on attenuating experimentally produced anger and anxiety. Specifically, we examined the assuaging of negative emotions through exposure to an image of a mother-infant embrace or a heterosexual couple embracing. Subjects seated at a computer terminal rated their affect (Pre Affect) using the Affect Adjective Checklist (AAC) then listened to two sets of intense two person conflicts. After the first conflict exposure they rated affect again (Post 1 AAC). Following the second exposure they saw a blank screen (control condition), pictures of everyday objects (distraction condition) or a photo of two people embracing (Secure Base Prime condition). They then reported emotions using the Post 2 AAC. Compared to either control or distraction subjects, Secure Base Prime (SBP) subjects reported significantly less anger and anxiety. These results were then replicated using an internet sample with control, SBP and two new controls: Smiling Man (to control for expression of positive affect) and Cold Mother (an unsmiling mother with infant). The SBP amelioration of anger and anxiety was replicated with the internet sample. No control groups produced this effect, which was generated only by a combination of positive affect in a physically embracing dyad. The results are discussed in terms of attachment theory and research on spreading activation. PMID:27606897

  16. Mo-Si-B-Based Coatings for Ceramic Base Substrates

    NASA Technical Reports Server (NTRS)

    Perepezko, John Harry (Inventor); Sakidja, Ridwan (Inventor); Ritt, Patrick (Inventor)

    2015-01-01

    Alumina-containing coatings based on molybdenum (Mo), silicon (Si), and boron (B) ("MoSiB coatings") that form protective, oxidation-resistant scales on ceramic substrate at high temperatures are provided. The protective scales comprise an aluminoborosilicate glass, and may additionally contain molybdenum. Two-stage deposition methods for forming the coatings are also provided.

  17. Factorization-based texture segmentation

    DOE PAGESBeta

    Yuan, Jiangye; Wang, Deliang; Cheriyadat, Anil M.

    2015-06-17

    This study introduces a factorization-based approach that efficiently segments textured images. We use local spectral histograms as features, and construct an M × N feature matrix using M-dimensional feature vectors in an N-pixel image. Based on the observation that each feature can be approximated by a linear combination of several representative features, we factor the feature matrix into two matrices-one consisting of the representative features and the other containing the weights of representative features at each pixel used for linear combination. The factorization method is based on singular value decomposition and nonnegative matrix factorization. The method uses local spectral histogramsmore » to discriminate region appearances in a computationally efficient way and at the same time accurately localizes region boundaries. Finally, the experiments conducted on public segmentation data sets show the promise of this simple yet powerful approach.« less

  18. Microcomputer logarithmic time base generator

    NASA Astrophysics Data System (ADS)

    Wills, L. J.; Ly, Nhan G.

    1985-11-01

    A new circuit is introduced to generate the logarithmic time base function with good resolution. By using a single-chip microcomputer with EPROM program storage, the circuitry is simplified and can be easily reproduced. The output function covers more than six decades of time and has 590 discrete points per decade with an accuracy of one discrete point per decade or ±0.16%. The design overcomes two well-known problems in using the logarithmic time base. First because the time increments are derived from a real-time register there is a precise reference for zero time, and second a series of time base interval marks are output for correctly calibrating the time axis.

  19. DEA based neonatal lung simulator

    NASA Astrophysics Data System (ADS)

    Schlatter, Samuel; Haemmerle, Enrico; Chang, Robin; O'Brien, Benjamin M.; Gisby, Todd; Anderson, Iain

    2011-04-01

    To reduce the likelihood of ventilator induced lung injury a neonatal lung simulator is developed based on Dielectric Elastomer Actuators (DEAs). DEAs are particularly suited for this application due to their natural like response as well as their self-sensing ability. By actively controlling the DEA, the pressure and volume inside the lung simulator can be controlled giving rise to active compliance control. Additionally the capacitance of the DEA can be used as a measurement of volume eliminating the integration errors that plague flow sensors. Based on simulations conducted with the FEA package ABAQUS and experimental data, the characteristics of the lung simulator were explored. A relationship between volume and capacitance was derived based on the self sensing of a bubble actuator. This was then used to calculate the compliance of the experimental bubble actuator. The current results are promising and show that mimicking a neonatal lung with DEAs may be possible.

  20. Materials properties data base computerization

    NASA Technical Reports Server (NTRS)

    Baur, R. G.; Donthnier, M. L.; Moran, M. C.; Mortman, I.; Pinter, R. S.

    1984-01-01

    Material property data plays a key role in the design of jet engine components. Consistency, accuracy and efficient use of material property data is of prime importance to the engineering community. The system conception, development, implementation, and future plans for computer software that captures the Material Properties Handbook into a scientific data base are described. The engineering community is given access to raw data and property curves, display of multiple curves for material evaluation and selection, direct access by design analysis computer programs, display of the material specification, and a historical repository for the material evolution. The impact of this activity includes significant productivity gains and cost reductions; all users have access to the same information nd provides consistent, rapid response to the needs of the engineering community. Future plans include incorporating the materials properties data base into a network environment to access information from other data bases and download information to engineering work stations.

  1. Coal Reserves Data Base report

    SciTech Connect

    Jones, R.W.; Glass, G.B.

    1991-12-05

    The Coal Reserves Data Base (CRDB) Program is a cooperative data base development program sponsored by the Energy Information Administration (EIA). The objective of the CRDB Program is to involve knowledgeable coal resource authorities from the major coal-bearing regions in EIA's effort to update the Nation's coal reserves data. This report describes one of two prototype studies to update State-level reserve estimates. The CRDB data are intended for use in coal supply analyses and to support analyses of policy and legislative issues. They will be available to both Government and non-Government analysts. The data also will be part of the information used to supply United States energy data for international data bases and for inquiries from private industry and the public. (VC)

  2. Brain plasticity-based therapeutics

    PubMed Central

    Merzenich, Michael M.; Van Vleet, Thomas M.; Nahum, Mor

    2014-01-01

    The primary objective of this review article is to summarize how the neuroscience of brain plasticity, exploiting new findings in fundamental, integrative and cognitive neuroscience, is changing the therapeutic landscape for professional communities addressing brain-based disorders and disease. After considering the neurological bases of training-driven neuroplasticity, we shall describe how this neuroscience-guided perspective distinguishes this new approach from (a) the more-behavioral, traditional clinical strategies of professional therapy practitioners, and (b) an even more widely applied pharmaceutical treatment model for neurological and psychiatric treatment domains. With that background, we shall argue that neuroplasticity-based treatments will be an important part of future best-treatment practices in neurological and psychiatric medicine. PMID:25018719

  3. Cellular-based preemption system

    NASA Technical Reports Server (NTRS)

    Bachelder, Aaron D. (Inventor)

    2011-01-01

    A cellular-based preemption system that uses existing cellular infrastructure to transmit preemption related data to allow safe passage of emergency vehicles through one or more intersections. A cellular unit in an emergency vehicle is used to generate position reports that are transmitted to the one or more intersections during an emergency response. Based on this position data, the one or more intersections calculate an estimated time of arrival (ETA) of the emergency vehicle, and transmit preemption commands to traffic signals at the intersections based on the calculated ETA. Additional techniques may be used for refining the position reports, ETA calculations, and the like. Such techniques include, without limitation, statistical preemption, map-matching, dead-reckoning, augmented navigation, and/or preemption optimization techniques, all of which are described in further detail in the above-referenced patent applications.

  4. Factorization-based texture segmentation

    SciTech Connect

    Yuan, Jiangye; Wang, Deliang; Cheriyadat, Anil M.

    2015-06-17

    This study introduces a factorization-based approach that efficiently segments textured images. We use local spectral histograms as features, and construct an M × N feature matrix using M-dimensional feature vectors in an N-pixel image. Based on the observation that each feature can be approximated by a linear combination of several representative features, we factor the feature matrix into two matrices-one consisting of the representative features and the other containing the weights of representative features at each pixel used for linear combination. The factorization method is based on singular value decomposition and nonnegative matrix factorization. The method uses local spectral histograms to discriminate region appearances in a computationally efficient way and at the same time accurately localizes region boundaries. Finally, the experiments conducted on public segmentation data sets show the promise of this simple yet powerful approach.

  5. Photoelectrochemical based direct conversion systems

    SciTech Connect

    Kocha, S.; Arent, D.; Peterson, M.

    1995-09-01

    The goal of this research is to develop a stable, cost effective, photoelectrochemical based system that will split water upon illumination, producing hydrogen and oxygen directly, using sunlight as the only energy input. This type of direct conversion system combines a photovoltaic material and an electrolyzer into a single monolithic device. We report on our studies of two multifunction multiphoton photoelectrochemical devices, one based on the ternary semiconductor gallium indium phosphide, (GaInP{sub 2}), and the other one based on amorphous silicon carbide. We also report on our studies of the solid state surface treatment of GaInP{sub 2} as well as our continuing effort to develop synthetic techniques for the attachment of transition metal complexes to the surface of semiconductor electrodes. All our surface studies are directed at controlling the interface energetics and forming stable catalytic surfaces.

  6. Developing Constraint-based Recommenders

    NASA Astrophysics Data System (ADS)

    Felfernig, Alexander; Friedrich, Gerhard; Jannach, Dietmar; Zanker, Markus

    Traditional recommendation approaches (content-based filtering [48] and collaborative filtering[40]) are well-suited for the recommendation of quality&taste products such as books, movies, or news. However, especially in the context of products such as cars, computers, appartments, or financial services those approaches are not the best choice (see also Chapter 11). For example, apartments are not bought very frequently which makes it rather infeasible to collect numerous ratings for one specific item (exactly such ratings are required by collaborative recommendation algorithms). Furthermore, users of recommender applications would not be satisfied with recommendations based on years-old item preferences (exactly such preferences would be exploited in this context by content-based filtering algorithms).

  7. Knowledge-based nursing diagnosis

    NASA Astrophysics Data System (ADS)

    Roy, Claudette; Hay, D. Robert

    1991-03-01

    Nursing diagnosis is an integral part of the nursing process and determines the interventions leading to outcomes for which the nurse is accountable. Diagnoses under the time constraints of modern nursing can benefit from a computer assist. A knowledge-based engineering approach was developed to address these problems. A number of problems were addressed during system design to make the system practical extended beyond capture of knowledge. The issues involved in implementing a professional knowledge base in a clinical setting are discussed. System functions, structure, interfaces, health care environment, and terminology and taxonomy are discussed. An integrated system concept from assessment through intervention and evaluation is outlined.

  8. Transistor-based interface circuitry

    DOEpatents

    Taubman, Matthew S.

    2004-02-24

    Among the embodiments of the present invention is an apparatus that includes a transistor, a servo device, and a current source. The servo device is operable to provide a common base mode of operation of the transistor by maintaining an approximately constant voltage level at the transistor base. The current source is operable to provide a bias current to the transistor. A first device provides an input signal to an electrical node positioned between the emitter of the transistor and the current source. A second device receives an output signal from the collector of the transistor.

  9. Transistor-based interface circuitry

    DOEpatents

    Taubman, Matthew S.

    2007-02-13

    Among the embodiments of the present invention is an apparatus that includes a transistor, a servo device, and a current source. The servo device is operable to provide a common base mode of operation of the transistor by maintaining an approximately constant voltage level at the transistor base. The current source is operable to provide a bias current to the transistor. A first device provides an input signal to an electrical node positioned between the emitter of the transistor and the current source. A second device receives an output signal from the collector of the transistor.

  10. Space construction base control system

    NASA Technical Reports Server (NTRS)

    1978-01-01

    Aspects of an attitude control system were studied and developed for a large space base that is structurally flexible and whose mass properties change rather dramatically during its orbital lifetime. Topics of discussion include the following: (1) space base orbital pointing and maneuvering; (2) angular momentum sizing of actuators; (3) momentum desaturation selection and sizing; (4) multilevel control technique applied to configuration one; (5) one-dimensional model simulation; (6) N-body discrete coordinate simulation; (7) structural analysis math model formulation; and (8) discussion of control problems and control methods.

  11. Case-based medical informatics

    PubMed Central

    Pantazi, Stefan V; Arocha, José F; Moehr, Jochen R

    2004-01-01

    Background The "applied" nature distinguishes applied sciences from theoretical sciences. To emphasize this distinction, we begin with a general, meta-level overview of the scientific endeavor. We introduce the knowledge spectrum and four interconnected modalities of knowledge. In addition to the traditional differentiation between implicit and explicit knowledge we outline the concepts of general and individual knowledge. We connect general knowledge with the "frame problem," a fundamental issue of artificial intelligence, and individual knowledge with another important paradigm of artificial intelligence, case-based reasoning, a method of individual knowledge processing that aims at solving new problems based on the solutions to similar past problems. We outline the fundamental differences between Medical Informatics and theoretical sciences and propose that Medical Informatics research should advance individual knowledge processing (case-based reasoning) and that natural language processing research is an important step towards this goal that may have ethical implications for patient-centered health medicine. Discussion We focus on fundamental aspects of decision-making, which connect human expertise with individual knowledge processing. We continue with a knowledge spectrum perspective on biomedical knowledge and conclude that case-based reasoning is the paradigm that can advance towards personalized healthcare and that can enable the education of patients and providers. We center the discussion on formal methods of knowledge representation around the frame problem. We propose a context-dependent view on the notion of "meaning" and advocate the need for case-based reasoning research and natural language processing. In the context of memory based knowledge processing, pattern recognition, comparison and analogy-making, we conclude that while humans seem to naturally support the case-based reasoning paradigm (memory of past experiences of problem-solving and

  12. School-based influenza immunization.

    PubMed

    Hull, Harry F; Frauendienst, Renee S; Gundersen, Margene L; Monsen, Susan M; Fishbein, Daniel B

    2008-08-12

    Annual influenza vaccination of schoolchildren will protect individual vaccines and, with high coverage, may protect entire communities. Because schoolchildren are more difficult to reach than preschoolers, school-based immunization programs may be needed to reach a high percentage of children. We offered free live, attenuated influenza vaccine to all healthy schoolchildren (K-12) in three Minnesota counties. Counties vaccinated from 33% to 58% of students. Overall, 41% of enrolled children were vaccinated. Elementary students were vaccinated at higher rates than older students. Administrative costs averaged $9.78 per dose delivered. School-based immunization programs offer the potential to achieve higher vaccination coverage of schoolchildren at modest cost. PMID:18577411

  13. Laser-based coatings removal

    SciTech Connect

    Freiwald, J.G.; Freiwald, D.A.

    1995-10-01

    Over the years as building and equipment surfaces became contaminated with low levels of uranium or plutonium dust, coats of paint were applied to stabilize the contaminants in place. Most of the earlier paint used was lead-based paint. More recently, various non-lead-based paints, such as two-part epoxy, are used. For D&D (decontamination and decommissioning), it is desirable to remove the paints or other coatings rather than having to tear down and dispose of the entire building. This report describes the use of pulse-repetetion laser systems for the removal of paints and coatings.

  14. Cluster identification based on correlations

    NASA Astrophysics Data System (ADS)

    Schulman, L. S.

    2012-04-01

    The problem addressed is the identification of cooperating agents based on correlations created as a result of the joint action of these and other agents. A systematic method for using correlations beyond second moments is developed. The technique is applied to a didactic example, the identification of alphabet letters based on correlations among the pixels used in an image of the letter. As in this example, agents can belong to more than one cluster. Moreover, the identification scheme does not require that the patterns be known ahead of time.

  15. Equientangled bases in arbitrary dimensions

    SciTech Connect

    Karimipour, V.; Memarzadeh, L.

    2006-01-15

    For the space of two identical systems of arbitrary dimensions, we introduce a continuous family of bases with the following properties: (i) the bases are orthonormal (ii) in each basis, all the states have the same values of entanglement, and (iii) they continuously interpolate between the product basis and the maximally entangled basis. The states thus constructed may find applications in many areas related to the quantum information science including quantum cryptography, optimal Bell tests, and the investigation of the enhancement of channel capacity due to entanglement.

  16. [Bone-based anchorage failure].

    PubMed

    Mer, Grégoria; Brezulier, Damien; Sorel, Olivier

    2016-03-01

    The aim of this article is to list the circumstances likely to give rise to failure of orthodontic temporary bone-supported anchorage and, hence, to attempt to define criteria for correct miniscrew usage. Our study was based on a review of the literature and analyses of clinical cases. Our findings show that, with a sound knowledge of the indications for screw selection and positioning and of the insertion protocols combined with a clear understanding of orthodontic mechanics, bone-based anchorage can henceforth provide orthodontists with an essential tool to enable formerly unachievable dental movements and to stabilize unwanted movements, thus making treatment both more reliable and more effective. PMID:27083224

  17. Reliability-based casing design

    SciTech Connect

    Maes, M.A.; Gulati, K.C.; Johnson, R.C.; McKenna, D.L.; Brand, P.R.; Lewis, D.B.

    1995-06-01

    The present paper describes the development of reliability-based design criteria for oil and/or gas well casing/tubing. The approach is based on the fundamental principles of limit state design. Limit states for tubulars are discussed and specific techniques for the stochastic modeling of loading and resistance variables are described. Zonation methods and calibration techniques are developed which are geared specifically to the characteristic tubular design for both hydrocarbon drilling and production applications. The application of quantitative risk analysis to the development of risk-consistent design criteria is shown to be a major and necessary step forward in achieving more economic tubular design.

  18. Laser-based coatings removal

    SciTech Connect

    Freiwald, J.G.; Freiwald, D.

    1995-12-01

    Over the years as building and equipment surfaces became contaminated with low levels of uranium or plutonium dust, coats of paint were applied to stabilize the contaminants in place. Most of the earlier paint used was lead-based paint. More recently, various non-lead-based paints, such as two-part epoxy, are used. For D & D (decontamination and decommissioning), it is desirable to remove the paints or other coatings rather than having to tear down and dispose of the entire building.

  19. Why internet-based education?

    PubMed Central

    Gernsbacher, Morton Ann

    2015-01-01

    This essay illustrates five ways that Internet-based higher education can capitalize on fundamental principles of learning. Internet-based education can enable better mastery through distributed (shorter, more frequent) practice rather than massed (longer, less frequent) practice; it can optimize performance because it allows students to learn at their peak time of their day; it can deepen memory because it requires cheat-proof assignments and tests; it can promote critical thinking because it necessitates intellectual winnowing and sifting; and it can enhance writing skills by requiring students to write frequently and for a broad audience. PMID:25653625

  20. Knowledge based jet engine diagnostics

    NASA Technical Reports Server (NTRS)

    Jellison, Timothy G.; Dehoff, Ronald L.

    1987-01-01

    A fielded expert system automates equipment fault isolation and recommends corrective maintenance action for Air Force jet engines. The knowledge based diagnostics tool was developed as an expert system interface to the Comprehensive Engine Management System, Increment IV (CEMS IV), the standard Air Force base level maintenance decision support system. XMAM (trademark), the Expert Maintenance Tool, automates procedures for troubleshooting equipment faults, provides a facility for interactive user training, and fits within a diagnostics information feedback loop to improve the troubleshooting and equipment maintenance processes. The application of expert diagnostics to the Air Force A-10A aircraft TF-34 engine equipped with the Turbine Engine Monitoring System (TEMS) is presented.

  1. Towards a lunar base programme

    NASA Technical Reports Server (NTRS)

    Duke, M. B.; Mendell, W. W.; Roberts, B. B.

    1985-01-01

    When the requisite technlogy exists, the U.S. political process will inevitably include lunar surface activities as a major space objective. This article examines a manned lunar base in terms of three distinct functions: the scientific investigation of the moon and its environment; development of the capability to use lunar resources for beneficial purposes throughout the earth-moon system; and conduct of R and D leading to a self-sufficient and self-supporting manned lunar base. Three scenarios are outlined with respect to each possible function.

  2. Base case and perturbation scenarios

    SciTech Connect

    Edmunds, T

    1998-10-01

    This report describes fourteen energy factors that could affect electricity markets in the future (demand, process, source mix, etc.). These fourteen factors are believed to have the most influence on the State's energy environment. A base case, or most probable, characterization is given for each of these fourteen factors over a twenty year time horizon. The base case characterization is derived from quantitative and qualitative information provided by State of California government agencies, where possible. Federal government databases are nsed where needed to supplement the California data. It is envisioned that a initial selection of issue areas will be based upon an evaluation of them under base case conditions. For most of the fourteen factors, the report identities possible perturbations from base case values or assumptions that may be used to construct additional scenarios. Only those perturbations that are plausible and would have a significant effect on energy markets are included in the table. The fourteen factors and potential perturbations of the factors are listed in Table 1.1. These perturbations can be combined to generate internally consist.ent. combinations of perturbations relative to the base case. For example, a low natural gas price perturbation should be combined with a high natural gas demand perturbation. The factor perturbations are based upon alternative quantitative forecasts provided by other institutions (the Department of Energy - Energy Information Administration in some cases), changes in assumptions that drive the quantitative forecasts, or changes in assumptions about the structure of the California energy markets. The perturbations are intended to be used for a qualitative reexamination of issue areas after an initial evaluation under the base case. The perturbation information would be used as a "tiebreaker;" to make decisions regarding those issue areas that were marginally accepted or rejected under the base case. Hf a

  3. Literature-Based Instruction: Reshaping the Curriculum.

    ERIC Educational Resources Information Center

    Raphael, Taffy E., Ed.; Au, Kathryn H., Ed.

    Making a case for the value of literature-based instruction, this book presents an overview of the extensive knowledge base supporting literature-based approaches to literacy instruction. It notes that literature-based instruction goes beyond simply changing the kinds of texts children read--also required in literature-based instruction are an…

  4. Content-based Instruction for African Languages.

    ERIC Educational Resources Information Center

    Moshi, Lioba

    2001-01-01

    Examines content-based instruction for African languages and considers Schleicher's (2000) expatiation of goal-based instruction for African languages. Focuses on the parameters for content-based instruction, the profile of a content-based instructional program, the nature of content-based instruction, the first steps for African languages, and…

  5. Watershed-based survey designs.

    PubMed

    Detenbeck, Naomi E; Cincotta, Dan; Denver, Judith M; Greenlee, Susan K; Olsen, Anthony R; Pitchford, Ann M

    2005-04-01

    Watershed-based sampling design and assessment tools help serve the multiple goals for water quality monitoring required under the Clean Water Act, including assessment of regional conditions to meet Section 305(b), identification of impaired water bodies or watersheds to meet Section 303(d), and development of empirical relationships between causes or sources of impairment and biological responses. Creation of GIS databases for hydrography, hydrologically corrected digital elevation models, and hydrologic derivatives such as watershed boundaries and upstream-downstream topology of subcatchments would provide a consistent seamless nationwide framework for these designs. The elements of a watershed-based sample framework can be represented either as a continuous infinite set defined by points along a linear stream network, or as a discrete set of watershed polygons. Watershed-based designs can be developed with existing probabilistic survey methods, including the use of unequal probability weighting, stratification, and two-stage frames for sampling. Case studies for monitoring of Atlantic Coastal Plain streams, West Virginia wadeable streams, and coastal Oregon streams illustrate three different approaches for selecting sites for watershed-based survey designs. PMID:15861987

  6. Task-Based Writing Instruction

    ERIC Educational Resources Information Center

    Bantis, Alexandros

    2010-01-01

    The purpose of this study was to investigate the impact of task-based writing instruction, a communicative language-teaching method, on second language acquisition and differentiation of instruction for English language learners during the independent work time instructional component of the Open Court Reading program. Through student-teacher…

  7. Problem Based Learning in Metaverse

    ERIC Educational Resources Information Center

    Barry, Dana M.; Kanematsu, Hideyuki; Fukumura, Yoshimi

    2010-01-01

    Problem Based Learning (PBL) is a powerful tool for both science and engineering education in the real world. Therefore, Japanese educators/researchers (with assistance from a US educator) carried out a pilot study to determine the effectiveness of using PBL activities in Metaverse. Their project was carried out by student teams from the US and…

  8. [Forced Oscillations of DNA Bases].

    PubMed

    Yakushevich, L V; Krasnobaeva, L A

    2016-01-01

    This paper presents the results of the studying of forced angular oscillations of the DNA bases with the help of the mathematical model consisting of two coupled nonlinear differential equations that take into account the effects of dissipation and the influence of an external periodic field. The calculation results are illustrated for sequence of gene encoding interferon alpha 17 (IFNA 17).

  9. Network-Based Management Procedures.

    ERIC Educational Resources Information Center

    Buckner, Allen L.

    Network-based management procedures serve as valuable aids in organizational management, achievement of objectives, problem solving, and decisionmaking. Network techniques especially applicable to educational management systems are the program evaluation and review technique (PERT) and the critical path method (CPM). Other network charting…

  10. Project-Based Module Development.

    ERIC Educational Resources Information Center

    Meel, R. M. van

    A project management design for modularizing higher education at open universities was developed and tested. Literature in the fields of project management and development of modular curriculum materials was reviewed and used as a basis for developing a project-based approach to the process of developing modules for self-instruction. According to…

  11. WATERSHED-BASED SURVEY DESIGNS

    EPA Science Inventory

    Water-based sampling design and assessment tools help serve the multiple goals for water quality monitoring required under the Clean Water Act, including assessment of regional conditions to meet Section 305(b), identification if impaired water bodies or watersheds to meet Sectio...

  12. LOGO Based Instruction in Geometry.

    ERIC Educational Resources Information Center

    Yusuf, Mian Muhammad

    The objective of this pretest-posttest Quasi-Experimental Design study was to determine the effects of LOGO Based Instruction (LBI) compared to instruction by teacher lecture and pencil-and-paper activities on: (1) students' understanding of the concepts of point, ray, line, and line segment; (2) students' attitudes toward learning geometry,…

  13. Expert and Knowledge Based Systems.

    ERIC Educational Resources Information Center

    Demaid, Adrian; Edwards, Lyndon

    1987-01-01

    Discusses the nature and current state of knowledge-based systems and expert systems. Describes an expert system from the viewpoints of a computer programmer and an applications expert. Addresses concerns related to materials selection and forecasts future developments in the teaching of materials engineering. (ML)

  14. Data Based Instruction in Reading

    ERIC Educational Resources Information Center

    Ediger, Marlow

    2010-01-01

    Data based instruction has received much attention in educational literature. It relates well to measurement driven teaching and learning. Data may come from several sources including mandated tests, district wide testing, formative and summative evaluations, as well as teacher written tests. Objective information is intended for use in data based…

  15. Cationic surfactants based on ferrocene

    SciTech Connect

    Pankratov, V.A.; Kucherova, N.L.; Abramzon, A.A.

    1988-07-20

    Quaternary ammonium salts based on ferrocene were synthesized and their surface active properties were studied as potential cationic surfactants and for uses including antiknock compounds. The salts were halide and nitrate derivatives of dimethylferrocenylmethylammonium and were prepared by aminomethylation of ferrocene. Chemical reaction yields, melting points, surface tension isotherms, and other characteristics were assessed.

  16. Competency Based Refresher Nurse Curriculum.

    ERIC Educational Resources Information Center

    Lombardo, Mary C.

    This competency-based course is designed to update the skills and knowledge of inactive nurses desiring to return to active practice. Focus of the course is on organizing and managing patient care using the nursing process; performing nursing procedures, including medication administration; and reintegrating oneself into the professional…

  17. Indigenous Community-Based Education.

    ERIC Educational Resources Information Center

    May, Stephen, Ed.

    After a long history as a tool of forced assimilation of indigenous populations, education is now a key arena in which indigenous peoples can reclaim and revalue their languages and cultures and thereby improve the academic success of indigenous students. Community-based education offers a means by which indigenous peoples can regain a measure of…

  18. Computer-Based Career Interventions.

    ERIC Educational Resources Information Center

    Mau, Wei-Cheng

    The possible utilities and limitations of computer-assisted career guidance systems (CACG) have been widely discussed although the effectiveness of CACG has not been systematically considered. This paper investigates the effectiveness of a theory-based CACG program, integrating Sequential Elimination and Expected Utility strategies. Three types of…

  19. Research-Based Teacher Education.

    ERIC Educational Resources Information Center

    Peters, William H.

    There is an emerging research base on teacher effectiveness that should be used in redesigning teacher education programs. However, the complexity of teaching and learning does not permit simple solutions to complex problems. Several studies, projects, and programs with the goal of conducting research for teacher education programs are discussed.…

  20. Computer Based Virtual Field Trips.

    ERIC Educational Resources Information Center

    Clark, Kenneth F.; Hosticka, Alice; Schriver, Martha; Bedell, Jackie

    This paper discusses computer based virtual field trips that use technologies commonly found in public schools in the United States. The discussion focuses on the advantages of both using and creating these field trips for an instructional situation. A virtual field trip to Cumberland Island National Seashore, St. Marys, Georgia is used as a point…

  1. [Forced Oscillations of DNA Bases].

    PubMed

    Yakushevich, L V; Krasnobaeva, L A

    2016-01-01

    This paper presents the results of the studying of forced angular oscillations of the DNA bases with the help of the mathematical model consisting of two coupled nonlinear differential equations that take into account the effects of dissipation and the influence of an external periodic field. The calculation results are illustrated for sequence of gene encoding interferon alpha 17 (IFNA 17). PMID:27192830

  2. Model-Based Safety Analysis

    NASA Technical Reports Server (NTRS)

    Joshi, Anjali; Heimdahl, Mats P. E.; Miller, Steven P.; Whalen, Mike W.

    2006-01-01

    System safety analysis techniques are well established and are used extensively during the design of safety-critical systems. Despite this, most of the techniques are highly subjective and dependent on the skill of the practitioner. Since these analyses are usually based on an informal system model, it is unlikely that they will be complete, consistent, and error free. In fact, the lack of precise models of the system architecture and its failure modes often forces the safety analysts to devote much of their effort to gathering architectural details about the system behavior from several sources and embedding this information in the safety artifacts such as the fault trees. This report describes Model-Based Safety Analysis, an approach in which the system and safety engineers share a common system model created using a model-based development process. By extending the system model with a fault model as well as relevant portions of the physical system to be controlled, automated support can be provided for much of the safety analysis. We believe that by using a common model for both system and safety engineering and automating parts of the safety analysis, we can both reduce the cost and improve the quality of the safety analysis. Here we present our vision of model-based safety analysis and discuss the advantages and challenges in making this approach practical.

  3. Data base management systems activities

    NASA Technical Reports Server (NTRS)

    1983-01-01

    The Data Management System-1100 is designed to operate in conjunction with the UNIVAC 1100 Series Operating System on any 1100 Series computer. DMS-1100 is divided into the following four major software components: (1) Data Definition Languages (DDL); (2) Data Management Routine (DMR); (3) Data Manipulation Languages (DML); and (4) Data Base Utilities (DBU). These software components are described in detail.

  4. Risk-based decisionmaking (Panel)

    SciTech Connect

    Smith, T.H.

    1995-12-31

    By means of a panel discussion and extensive audience interaction, explore the current challenges and progress to date in applying risk considerations to decisionmaking related to low-level waste. This topic is especially timely because of the proposed legislation pertaining to risk-based decisionmaking and because of the increased emphasis placed on radiological performance assessments of low-level waste disposal.

  5. ANN - based distribution system reconfiguration

    SciTech Connect

    Momoh, J.A.; Wang, Yanchun; Rizy, D.T.

    1997-08-01

    This paper describes an Artificial Neural Network (ANN) - based distribution system reconfiguration scheme to reduce system loss. The ANN is trained for different load levels and different network topologies. The proposed scheme has been tested using a 38 - bus distribution system. The results are very promising.

  6. RF MEMS Based Reconfigurable Antennas

    NASA Technical Reports Server (NTRS)

    Simons, Rainee N.

    2004-01-01

    The presentation will first of all address the advantages of RF MEMS circuit in antenna applications and also the need for electronically reconfigurable antennas. Next, discuss some of the recent examples of RF MEMS based reconfigurable microstrip antennas. Finally, conclude the talk with a summary of MEMS antenna performance.

  7. Water-based cleaning fundamentals

    SciTech Connect

    Harding, W.B.

    1991-12-01

    A basic description of water-based alkaline cleaning is presented, The nature of soils is described. The compositions of conventional cleaning compounds are given with descriptions of the functions of the ingredients. The mechanisms by which soil is removed are explained. The degrees of cleanliness required, along with the influence of the material being cleaned, are discussed. Tests for cleanliness are described.

  8. Index to Computer Based Learning.

    ERIC Educational Resources Information Center

    Hoye, Robert E., Ed.; Wang, Anastasia C., Ed.

    The computer-based programs and projects described in this index are listed under 98 different subject matter fields. Descrptions of programs include information on: subject field, program name and number, author, source, the program's curriculum content, prerequisites, level of instruction, type of student for which it is intended, total hours of…

  9. Video-based crowd synthesis.

    PubMed

    Flagg, Matthew; Rehg, James M

    2013-11-01

    As a controllable medium, video-realistic crowds are important for creating the illusion of a populated reality in special effects, games, and architectural visualization. While recent progress in simulation and motion captured-based techniques for crowd synthesis has focused on natural macroscale behavior, this paper addresses the complementary problem of synthesizing crowds with realistic microscale behavior and appearance. Example-based synthesis methods such as video textures are an appealing alternative to conventional model-based methods, but current techniques are unable to represent and satisfy constraints between video sprites and the scene. This paper describes how to synthesize crowds by segmenting pedestrians from input videos of natural crowds and optimally placing them into an output video while satisfying environmental constraints imposed by the scene. We introduce crowd tubes, a representation of video objects designed to compose a crowd of video billboards while avoiding collisions between static and dynamic obstacles. The approach consists of representing crowd tube samples and constraint violations with a conflict graph. The maximal independent set yields a dense constraint-satisfying crowd composition. We present a prototype system for the capture, analysis, synthesis, and control of video-based crowds. Several results demonstrate the system's ability to generate videos of crowds which exhibit a variety of natural behaviors. PMID:24029912

  10. School Centered Evidence Based Accountability

    ERIC Educational Resources Information Center

    Milligan, Charles

    2015-01-01

    Achievement scores drive much of the effort in today's accountability system, however, there is much more that occurs in every school, every day. School Centered Evidence Based Accountability can be used from micro to macro giving School Boards and Administration a process for monitoring the results of the entire school operation effectively and…

  11. Watershed-based survey designs

    USGS Publications Warehouse

    Detenbeck, N.E.; Cincotta, D.; Denver, J.M.; Greenlee, S.K.; Olsen, A.R.; Pitchford, A.M.

    2005-01-01

    Watershed-based sampling design and assessment tools help serve the multiple goals for water quality monitoring required under the Clean Water Act, including assessment of regional conditions to meet Section 305(b), identification of impaired water bodies or watersheds to meet Section 303(d), and development of empirical relationships between causes or sources of impairment and biological responses. Creation of GIS databases for hydrography, hydrologically corrected digital elevation models, and hydrologic derivatives such as watershed boundaries and upstream-downstream topology of subcatchments would provide a consistent seamless nationwide framework for these designs. The elements of a watershed-based sample framework can be represented either as a continuous infinite set defined by points along a linear stream network, or as a discrete set of watershed polygons. Watershed-based designs can be developed with existing probabilistic survey methods, including the use of unequal probability weighting, stratification, and two-stage frames for sampling. Case studies for monitoring of Atlantic Coastal Plain streams, West Virginia wadeable streams, and coastal Oregon streams illustrate three different approaches for selecting sites for watershed-based survey designs. ?? Springer Science + Business Media, Inc. 2005.

  12. Competency-Based Behavioral Interviewing

    ERIC Educational Resources Information Center

    Garrigues, Sarah M.

    2012-01-01

    Directors too often rely primarily on their hunches to guide them in employee selection. But what if their hunches are only 30 percent correct? Potentially, one bad hire could be responsible for a noticeable decrease in enrollment and, as a result, the school's net revenue. In this article, the author discusses the Competency-Based Behavioral…

  13. Faceted Taxonomy-Based Sources

    NASA Astrophysics Data System (ADS)

    Tzitzikas, Yannis

    The objective of this chapter is to explain the underlying mathematical structure of faceted taxonomy-based sources and to provide some common notions and notations that are used in some parts of the book. Subsequently, and on the basis of the introduced formalism, this chapter describes the interaction between a user and an information source that supports dynamic taxonomies and faceted search.

  14. MTA Computer Based Evaluation System.

    ERIC Educational Resources Information Center

    Brenner, Lisa P.; And Others

    The MTA PLATO-based evaluation system, which has been implemented by a consortium of schools of medical technology, is designed to be general-purpose, modular, data-driven, and interactive, and to accommodate other national and local item banks. The system provides a comprehensive interactive item-banking system in conjunction with online student…

  15. Population Education: A Knowledge Base.

    ERIC Educational Resources Information Center

    Jacobson, Willard J.

    To aid junior high and high school educators and curriculum planners as they develop population education programs, the book provides an overview of the population education knowledge base. In addition, it suggests learning activities, discussion questions, and background information which can be integrated into courses dealing with population,…

  16. Colour Mixing Based on Daylight

    ERIC Educational Resources Information Center

    Meyn, Jan-Peter

    2008-01-01

    Colour science is based on the sensation of monochromatic light. In contrast to that, surface colours are caused by reflection of wide sections of the daylight spectrum. Non-spectral colours like magenta and purple appear homologous to colours with spectral hue, if the approach of mixing monochromatic light is abandoned. It is shown that a large…

  17. Ground-based IRCM testing

    NASA Astrophysics Data System (ADS)

    Greer, Derek; Owen, Mark

    2010-04-01

    Recent advances in the ability to perform comprehensive ground based Infrared Countermeasure (IRCM) testing have the capability to fill the Test and Evaluation (T&E) gaps for existing and future weapons system acquisition. IRCM testing has historically been dominated and in a manner limited by expensive live fire testing requirements. While live fire testing is a vital part of IRCM T&E, next generation technological developments now enable closed-loop, ground-based IRCM testing to provide valuable complementary test data at a much lower cost. The high cost and limited assets that have prevented live fire and flight testing from providing a thorough hardware based data set required for previous T&E analysis is no longer an issue. In the past, traditional physics based digital system model (DSM) analysis has been utilized to augment the IRCM data sets to make them statistically significant. While DSM is a useful tool in the development of IRCM systems, the newly developed installed system testing utilizing a hardware-in-the-loop construct provides for an enhanced level of fidelity and assurance that the systems will meet the warfighter's needs. The goal of the newly developed test technologies is to develop a statistical significant data set utilizing hardware-in-the-loop at a significantly lower cost than historical methods.

  18. Graphite-based photovoltaic cells

    DOEpatents

    Lagally, Max; Liu, Feng

    2010-12-28

    The present invention uses lithographically patterned graphite stacks as the basic building elements of an efficient and economical photovoltaic cell. The basic design of the graphite-based photovoltaic cells includes a plurality of spatially separated graphite stacks, each comprising a plurality of vertically stacked, semiconducting graphene sheets (carbon nanoribbons) bridging electrically conductive contacts.

  19. Budgeting Based on Student Needs

    ERIC Educational Resources Information Center

    Willis, Jason; Hill, Matt

    2011-01-01

    School finance reform has become a key component for transforming public schools in the United States. Over the last decade, a growing number of districts have turned to an approach known by different names--student-based budgeting, weighted student funding and fair student funding, among others--in which budgets are allocated to schools in…

  20. Video-based crowd synthesis.

    PubMed

    Flagg, Matthew; Rehg, James M

    2013-11-01

    As a controllable medium, video-realistic crowds are important for creating the illusion of a populated reality in special effects, games, and architectural visualization. While recent progress in simulation and motion captured-based techniques for crowd synthesis has focused on natural macroscale behavior, this paper addresses the complementary problem of synthesizing crowds with realistic microscale behavior and appearance. Example-based synthesis methods such as video textures are an appealing alternative to conventional model-based methods, but current techniques are unable to represent and satisfy constraints between video sprites and the scene. This paper describes how to synthesize crowds by segmenting pedestrians from input videos of natural crowds and optimally placing them into an output video while satisfying environmental constraints imposed by the scene. We introduce crowd tubes, a representation of video objects designed to compose a crowd of video billboards while avoiding collisions between static and dynamic obstacles. The approach consists of representing crowd tube samples and constraint violations with a conflict graph. The maximal independent set yields a dense constraint-satisfying crowd composition. We present a prototype system for the capture, analysis, synthesis, and control of video-based crowds. Several results demonstrate the system's ability to generate videos of crowds which exhibit a variety of natural behaviors.