Science.gov

Sample records for coelicolor development demonstrates

  1. Plasticity of Streptomyces coelicolor Membrane Composition Under Different Growth Conditions and During Development

    PubMed Central

    Sandoval-Calderón, Mario; Nguyen, Don D.; Kapono, Clifford A.; Herron, Paul; Dorrestein, Pieter C.; Sohlenkamp, Christian

    2015-01-01

    Streptomyces coelicolor is a model actinomycete that is well known for the diversity of its secondary metabolism and its complex life cycle. As a soil inhabitant, it is exposed to heterogeneous and frequently changing environmental circumstances. In the present work, we studied the effect of diverse growth conditions and phosphate depletion on its lipid profile and the relationship between membrane lipid composition and development in S. coelicolor. The lipid profile from cultures grown on solid media, which is closer to the natural habitat of this microorganism, does not resemble the previously reported lipid composition from liquid grown cultures of S. coelicolor. Wide variations were also observed across different media, growth phases, and developmental stages indicating active membrane remodeling. Ornithine lipids (OL) are phosphorus-free polar lipids that were accumulated mainly during sporulation stages, but were also major components of the membrane under phosphorus limitation. In contrast, phosphatidylethanolamine, which had been reported as one of the major polar lipids in the genus Streptomyces, is almost absent under these conditions. We identified one of the genes responsible for the synthesis of OL (SCO0921) and found that its inactivation causes the absence of OL, precocious morphological development and actinorhodin production. Our observations indicate a remarkable plasticity of the membrane composition in this bacterial species, reveal a higher metabolic complexity than expected, and suggest a relationship between cytoplasmic membrane components and the differentiation programs in S. coelicolor. PMID:26733994

  2. Pleiotropic effects of cAMP on germination, antibiotic biosynthesis and morphological development in Streptomyces coelicolor.

    PubMed

    Süsstrunk, U; Pidoux, J; Taubert, S; Ullmann, A; Thompson, C J

    1998-10-01

    In wild-type Streptomyces coelicolor MT1110 cultures, cyclic adenosine 3',5' monophosphate (cAMP) was synthesized throughout the developmental programme with peaks of accumulation both during germination and later when aerial mycelium and actinorhodin were being produced. Construction and characterization of an adenylate cyclase disruption mutant (BZ1) demonstrated that cAMP facilitated these developmental processes. Although pulse-labelling experiments showed that a similar germination process was initiated in BZ1 and MT1110, germ-tube emergence was severely delayed in BZ1 and never occurred in more than 85% of the spores. Studies of growth and development on solid glucose minimal medium (SMMS, buffered or unbuffered) showed that MT1110 and BZ1 produced acid during the first rapid growth phase, which generated substrate mycelium. Thereafter, on unbuffered SMMS, only MT1110 resumed growth and produced aerial mycelium by switching to an alternative metabolism that neutralized its medium, probably by reincorporating and metabolizing extracellular acids. BZ1 was not able to neutralize its medium or produce aerial mycelium on unbuffered SMMS; these defects were suppressed by high concentrations (>1 mM) of cAMP during early growth or on buffered medium. Other developmental mutants (bldA, bldB, bldC, bldD, bldG) also irreversibly acidified this medium. However, these bald mutants were not suppressed by exogenous cAMP or neutralizing buffer. BZ1 also differentiated when it was cultured in close proximity to MT1110, a property observed in cross-feeding experiments between bald mutants and commonly thought to reflect diffusion of a discrete positively acting signalling molecule. In this case, MT1110 generated a more neutral pH environment that allowed BZ1 to reinitiate growth and form aerial mycelium. The fact that actinorhodin synthesis could be induced by concentrations of cAMP (< 20 microM) found in the medium of MT1110 cultures, suggested that it may serve as a

  3. Transcriptomic Analysis of Liquid Non-Sporulating Streptomyces coelicolor Cultures Demonstrates the Existence of a Complex Differentiation Comparable to That Occurring in Solid Sporulating Cultures

    PubMed Central

    Yagüe, Paula; Rodríguez-García, Antonio; López-García, María Teresa; Rioseras, Beatriz; Martín, Juan Francisco; Sánchez, Jesús; Manteca, Angel

    2014-01-01

    Streptomyces species produce many clinically relevant secondary metabolites and exhibit a complex development that includes hyphal differentiation and sporulation in solid cultures. Industrial fermentations are usually performed in liquid cultures, conditions in which Streptomyces strains generally do not sporulate, and it was traditionally assumed that no differentiation took place. The aim of this work was to compare the transcriptomes of S. coelicolor growing in liquid and solid cultures, deepening the knowledge of Streptomyces differentiation. Microarrays demonstrated that gene expression in liquid and solid cultures were comparable and data indicated that physiological differentiation was similar for both conditions. Eighty-six percent of all transcripts showed similar abundances in liquid and solid cultures, such as those involved in the biosynthesis of actinorhodin (actVA, actII-4) and undecylprodigiosin (redF); activation of secondary metabolism (absR1, ndsA); genes regulating hydrophobic cover formation (aerial mycelium) (bldB, bldC, bldM, bldN, sapA, chpC, chpD, chpE, chpH, ramA, ramC, ramS); and even some genes regulating early stages of sporulation (wblA, whiG, whiH, whiJ). The two most important differences between transcriptomes from liquid and solid cultures were: first, genes related to secondary metabolite biosynthesis (CDA, CPK, coelichelin, desferrioxamine clusters) were highly up-regulated in liquid but not in solid cultures; and second, genes involved in the final stages of hydrophobic cover/spore maturation (chpF, rdlA, whiE, sfr) were up-regulated in solid but not in liquid cultures. New information was also provided for several non-characterized genes differentially expressed in liquid and solid cultures which might be regulating, at least in part, the metabolic and developmental differences observed between liquid and solid cultures. PMID:24466012

  4. Transcriptomic analysis of liquid non-sporulating Streptomyces coelicolor cultures demonstrates the existence of a complex differentiation comparable to that occurring in solid sporulating cultures.

    PubMed

    Yagüe, Paula; Rodríguez-García, Antonio; López-García, María Teresa; Rioseras, Beatriz; Martín, Juan Francisco; Sánchez, Jesús; Manteca, Angel

    2014-01-01

    Streptomyces species produce many clinically relevant secondary metabolites and exhibit a complex development that includes hyphal differentiation and sporulation in solid cultures. Industrial fermentations are usually performed in liquid cultures, conditions in which Streptomyces strains generally do not sporulate, and it was traditionally assumed that no differentiation took place. The aim of this work was to compare the transcriptomes of S. coelicolor growing in liquid and solid cultures, deepening the knowledge of Streptomyces differentiation. Microarrays demonstrated that gene expression in liquid and solid cultures were comparable and data indicated that physiological differentiation was similar for both conditions. Eighty-six percent of all transcripts showed similar abundances in liquid and solid cultures, such as those involved in the biosynthesis of actinorhodin (actVA, actII-4) and undecylprodigiosin (redF); activation of secondary metabolism (absR1, ndsA); genes regulating hydrophobic cover formation (aerial mycelium) (bldB, bldC, bldM, bldN, sapA, chpC, chpD, chpE, chpH, ramA, ramC, ramS); and even some genes regulating early stages of sporulation (wblA, whiG, whiH, whiJ). The two most important differences between transcriptomes from liquid and solid cultures were: first, genes related to secondary metabolite biosynthesis (CDA, CPK, coelichelin, desferrioxamine clusters) were highly up-regulated in liquid but not in solid cultures; and second, genes involved in the final stages of hydrophobic cover/spore maturation (chpF, rdlA, whiE, sfr) were up-regulated in solid but not in liquid cultures. New information was also provided for several non-characterized genes differentially expressed in liquid and solid cultures which might be regulating, at least in part, the metabolic and developmental differences observed between liquid and solid cultures.

  5. Exploration Technology Development & Demonstration

    NASA Video Gallery

    Chris Moore delivers a presentation from the Exploration Technology Development & Demonstration (ETDD) study team on May 25, 2010, at the NASA Exploration Enterprise Workshop held in Galveston, TX....

  6. Genes essential for morphological development and antibiotic production in Streptomyces coelicolor are targets of BldD during vegetative growth.

    PubMed

    den Hengst, Chris D; Tran, Ngat T; Bibb, Maureen J; Chandra, Govind; Leskiw, Brenda K; Buttner, Mark J

    2010-10-01

    BldD is a transcriptional regulator essential for morphological development and antibiotic production in Streptomyces coelicolor. Here we identify the BldD regulon by means of chromatin immunoprecipitation-microarray analysis (ChIP-chip). The BldD regulon encompasses ~167 transcriptional units, of which more than 20 are known to play important roles in development (e.g. bldA, bldC, bldH/adpA, bldM, bldN, ssgA, ssgB, ftsZ, whiB, whiG, smeA-ssfA) and/or secondary metabolism (e.g. nsdA, cvn9, bldA, bldC, leuA). Strikingly, 42 BldD target genes (~25% of the regulon) encode regulatory proteins, stressing the central, pleiotropic role of BldD. Almost all BldD binding sites identified by ChIP-chip are present in the promoters of the target genes. An exception is the tRNA gene bldA, where BldD binds within the region encoding the primary transcript, immediately downstream of the position corresponding to the processed, mature 3 end of the tRNA. Through gene overexpression, we identified a novel BldD target gene (cdgA) that influences differentiation and antibiotic production. cdgA encodes a GGDEF domain protein, implicating c-di-GMP in the regulation of Streptomyces development. Sequence analysis of the upstream regions of the complete regulon identified a 15 bp inverted repeat that functions as a high-affinity binding site for BldD, as was shown by electrophoretic mobility shift assays and DNase I footprinting analysis. High-scoring copies of the BldD binding site were found at relevant positions in the genomes of other bacteria containing a BldD homologue, suggesting the role of BldD is conserved in sporulating actinomycetes.

  7. Interactions between Streptomyces coelicolor and Bacillus subtilis: Role of Surfactants in Raising Aerial Structures

    PubMed Central

    Straight, Paul D.; Willey, Joanne M.; Kolter, Roberto

    2006-01-01

    Using mixed-species cultures, we have undertaken a study of interactions between two common spore-forming soil bacteria, Bacillus subtilis and Streptomyces coelicolor. Our experiments demonstrate that the development of aerial hyphae and spores by S. coelicolor is inhibited by surfactin, a lipopeptide surfactant produced by B. subtilis. Current models of aerial development by sporulating bacteria and fungi postulate a role for surfactants in reducing surface tension at air-liquid interfaces, thereby removing the major barrier to aerial growth. S. coelicolor produces SapB, an amphipathic peptide that is surface active and required for aerial growth on certain media. Loss of aerial hyphae in developmental mutants can be rescued by addition of purified SapB. While a surfactant from a fungus can substitute for SapB in a mutant that lacks aerial hyphae, not all surfactants have this effect. We show that surfactin is required for formation of aerial structures on the surface of B. subtilis colonies. However, in contrast to this positive role, our experiments reveal that surfactin acts antagonistically by arresting S. coelicolor aerial development and causing altered expression of developmental genes. Our observations support the idea that surfactants function specifically for a given organism regardless of their shared ability to reduce surface tension. Production of surfactants with antagonistic activity could provide a powerful competitive advantage during surface colonization and in competition for resources. PMID:16788200

  8. Identification of glucose kinase-dependent and -independent pathways for carbon control of primary metabolism, development and antibiotic production in Streptomyces coelicolor by quantitative proteomics.

    PubMed

    Gubbens, Jacob; Janus, Marleen; Florea, Bogdan I; Overkleeft, Herman S; van Wezel, Gilles P

    2012-12-01

    Members of the soil-dwelling prokaryotic genus Streptomyces are indispensable for the recycling of complex polysaccharides, and produce a wide range of natural products. Nutrient availability is a major determinant for the switch to development and antibiotic production in streptomycetes. Carbon catabolite repression (CCR), a main signalling pathway underlying this phenomenon, was so far considered fully dependent on the glycolytic enzyme glucose kinase (Glk). Here we provide evidence of a novel Glk-independent pathway in Streptomyces coelicolor, using advanced proteomics that allowed the comparison of the expression of some 2000 proteins, including virtually all enzymes for central metabolism. While CCR and inducer exclusion of enzymes for primary and secondary metabolism and precursor supply for natural products is mostly mediated via Glk, enzymes for the urea cycle, as well as for biosynthesis of the γ-butyrolactone Scb1 and the responsive cryptic polyketide Cpk are subject to Glk-independent CCR. Deletion of glkA led to strong downregulation of biosynthetic proteins for prodigionins and calcium-dependent antibiotic (CDA) in mannitol-grown cultures. Repression of bldB, bldN, and its target bldM may explain the poor development of S. coelicolor on solid-grown cultures containing glucose. A new model for carbon catabolite repression in streptomycetes is presented.

  9. ssgA Is Essential for Sporulation of Streptomyces coelicolor A3(2) and Affects Hyphal Development by Stimulating Septum Formation

    PubMed Central

    van Wezel, Gilles P.; van der Meulen, Jannes; Kawamoto, Shinichi; Luiten, Ruud G. M.; Koerten, Henk K.; Kraal, Barend

    2000-01-01

    The role of ssgA in cell division and development of streptomycetes was analyzed. An ssgA null mutant of Streptomyces coelicolor produced aerial hyphae but failed to sporulate, and ssgA can therefore be regarded as a novel whi gene. In addition to the morphological changes, antibiotic production was also disturbed, with strongly reduced actinorhodin production. These defects could be complemented by plasmid-borne ssgA. In the wild-type strain, transcription of ssgA was induced by nutritional shift-down and was shown to be linked to that of the upstream-located gene ssgR, which belongs to the family of iclR-type transcriptional regulator genes. Analysis of mycelium harvested from liquid-grown cultures by transmission electron microscopy showed that septum formation had strongly increased in ssgA-overexpressing strains in comparison to wild-type S. coelicolor and that spore-like compartments were produced at high frequency. Furthermore, the hyphae were significantly wider and contained irregular and often extremely thick septa. These data underline the important role for ssgA in Streptomyces cell division. PMID:11004161

  10. Functional Analysis of the N-Acetylglucosamine Metabolic Genes of Streptomyces coelicolor and Role in Control of Development and Antibiotic Production

    PubMed Central

    Świątek, Magdalena A.; Tenconi, Elodie; Rigali, Sébastien

    2012-01-01

    N-Acetylglucosamine, the monomer of chitin, is a favored carbon and nitrogen source for streptomycetes. Its intracellular catabolism requires the combined actions of the N-acetylglucosamine-6-phosphate (GlcNAc-6P) deacetylase NagA and the glucosamine-6-phosphate (GlcN-6P) deaminase/isomerase NagB. GlcNAc acts as a signaling molecule in the DasR-mediated nutrient sensing system, activating development and antibiotic production under poor growth conditions (famine) and blocking these processes under rich conditions (feast). In order to understand how a single nutrient can deliver opposite information according to the nutritional context, we carried out a mutational analysis of the nag metabolic genes nagA, nagB, and nagK. Here we show that the nag genes are part of the DasR regulon in Streptomyces coelicolor, which explains their transcriptional induction by GlcNAc. Most likely as the result of the intracellular accumulation of GlcN-6P, nagB deletion mutants fail to grow in the presence of GlcNAc. This toxicity can be alleviated by the additional deletion of nagA. We recently showed that in S. coelicolor, GlcNAc is internalized as GlcNAc-6P via the phosphoenolpyruvate-dependent sugar phosphotransferase system (PTS). Considering the relevance of GlcNAc for the control of antibiotic production, improved insight into GlcNAc metabolism in Streptomyces may provide new leads toward biotechnological applications. PMID:22194457

  11. Mycelium differentiation and development of Streptomyces coelicolor in lab-scale bioreactors: programmed cell death, differentiation, and lysis are closely linked to undecylprodigiosin and actinorhodin production.

    PubMed

    Rioseras, Beatriz; López-García, María Teresa; Yagüe, Paula; Sánchez, Jesús; Manteca, Angel

    2014-01-01

    Streptomycetes are mycelium-forming bacteria that produce two thirds of clinically relevant secondary metabolites. Secondary metabolite production is activated at specific developmental stages of Streptomyces life cycle. Despite this, Streptomyces differentiation in industrial bioreactors tends to be underestimated and the most important parameters managed are only indirectly related to differentiation: modifications to the culture media, optimization of productive strains by random or directed mutagenesis, analysis of biophysical parameters, etc. In this work the relationship between differentiation and antibiotic production in lab-scale bioreactors was defined. Streptomyces coelicolor was used as a model strain. Morphological differentiation was comparable to that occurring during pre-sporulation stages in solid cultures: an initial compartmentalized mycelium suffers a programmed cell death, and remaining viable segments then differentiate to a second multinucleated antibiotic-producing mycelium. Differentiation was demonstrated to be one of the keys to interpreting biophysical fermentation parameters and to rationalizing the optimization of secondary metabolite production in bioreactors.

  12. Mycelium differentiation and development of Streptomyces coelicolor in lab-scale bioreactors: Programmed cell death, differentiation, and lysis are closely linked to undecylprodigiosin and actinorhodin production

    PubMed Central

    Rioseras, Beatriz; López-García, María Teresa; Yagüe, Paula; Sánchez, Jesús; Manteca, Ángel

    2013-01-01

    Streptomycetes are mycelium-forming bacteria that produce two thirds of clinically relevant secondary metabolites. Secondary metabolite production is activated at specific developmental stages of Streptomyces life cycle. Despite this, Streptomyces differentiation in industrial bioreactors tends to be underestimated and the most important parameters managed are only indirectly related to differentiation: modifications to the culture media, optimization of productive strains by random or directed mutagenesis, analysis of biophysical parameters, etc. In this work the relationship between differentiation and antibiotic production in lab-scale bioreactors was defined. Streptomyces coelicolor was used as a model strain. Morphological differentiation was comparable to that occurring during pre-sporulation stages in solid cultures: an initial compartmentalized mycelium suffers a programmed cell death, and remaining viable segments then differentiate to a second multinucleated antibiotic-producing mycelium. Differentiation was demonstrated to be one of the keys to interpreting biophysical fermentation parameters and to rationalizing the optimization of secondary metabolite production in bioreactors. PMID:24240146

  13. A Laterally Acquired Galactose Oxidase-Like Gene Is Required for Aerial Development during Osmotic Stress in Streptomyces coelicolor

    PubMed Central

    Liman, Recep; Facey, Paul D.; van Keulen, Geertje; Dyson, Paul J.; Del Sol, Ricardo

    2013-01-01

    Phylogenetic reconstruction revealed that most Actinobacterial orthologs of S. coelicolor SCO2837, encoding a metal-dependent galactose oxidase-like protein, are found within Streptomyces and were probably acquired by horizontal gene transfer from fungi. Disruption of SCO2837 (glxA) caused a conditional bld phenotype that could not be reversed by extracellular complementation. Studies aimed at characterising the regulation of expression of glxA showed that it is not a target for other bld genes. We provide evidence that glxA is required for osmotic adaptation, although independently from the known osmotic stress response element SigB. glxA has been predicted to be part of an operon with the transcription unit comprising the upstream cslA gene and glxA. However, both phenotypic and expression studies indicate that it is also expressed from an independent promoter region internal to cslA. GlxA displays an in situ localisation pattern similar to that one observed for CslA at hyphal tips, but localisation of the former is independent of the latter. The functional role of GlxA in relation to CslA is discussed. PMID:23326581

  14. Phage-mediated cloning of bldA, a region involved in Streptomyces coelicolor morphological development, and its analysis by genetic complementation.

    PubMed Central

    Piret, J M; Chater, K F

    1985-01-01

    Streptomyces coelicolor bald (bld) mutants form colonies of vegetative substrate mycelium, but do not develop aerial hyphae or spore chains. The bldA strains form none of the four antibiotics known to be produced by the parent strain. With a vector derived from the temperate bacteriophage phi C31, a 5.6-kilobase fragment of wildtype DNA was cloned which restored sporulation to five independent bldA mutants when lysogenized with the recombinant phage. The cloned gene(s) was dominant over the mutant alleles. Phage integration by recombination of the cloned bldA+ DNA with the bldA region of each mutant produced mainly sporulating colonies, presumably heterozygous bldA+/bldA partial diploids for the insert DNA. However, a minority of these primary transductants were bald and were apparently homozygous bldA/bldA mutant partial diploids, formed by some homogenetization process. The phages released from the bald lysogens carried bldA mutations and were used to show that bldA+ sequences had been cloned and that fine mapping of the region could be performed. Images PMID:2993254

  15. The SmpB-tmRNA tagging system plays important roles in Streptomyces coelicolor growth and development.

    PubMed

    Yang, Chunzhong; Glover, John R

    2009-01-01

    The ssrA gene encodes tmRNA that, together with a specialized tmRNA-binding protein, SmpB, forms part of a ribonucleoprotein complex, provides a template for the resumption of translation elongation, subsequent termination and recycling of stalled ribosomes. In addition, the mRNA-like domain of tmRNA encodes a peptide that tags polypeptides derived from stalled ribosomes for degradation. Streptomyces are unique bacteria that undergo a developmental cycle culminating at sporulation that is at least partly controlled at the level of translation elongation by the abundance of a rare tRNA that decodes UUA codons found in a relatively small number of open reading frames prompting us to examine the role of tmRNA in S. coelicolor. Using a temperature sensitive replicon, we found that the ssrA gene could be disrupted only in cells with an extra-copy wild type gene but not in wild type cells or cells with an extra-copy mutant tmRNA (tmRNA(DD)) encoding a degradation-resistant tag. A cosmid-based gene replacement method that does not include a high temperature step enabled us to disrupt both the ssrA and smpB genes separately and at the same time suggesting that the tmRNA tagging system may be required for cell survival under high temperature. Indeed, mutant cells show growth and sporulation defects at high temperature and under optimal culture conditions. Interestingly, even though these defects can be completely restored by wild type genes, the DeltassrA strain was only partially corrected by tmRNA(DD). In addition, wildtype tmRNA can restore the hygromycin-resistance to DeltassrA cells while tmRNA(DD) failed to do so suggesting that degradation of aberrant peptides is important for antibiotic resistance. Overall, these results suggest that the tmRNA tagging system plays important roles during Streptomyces growth and sporulation under both normal and stress conditions.

  16. The SmpB-tmRNA Tagging System Plays Important Roles in Streptomyces coelicolor Growth and Development

    PubMed Central

    Yang, Chunzhong; Glover, John R.

    2009-01-01

    The ssrA gene encodes tmRNA that, together with a specialized tmRNA-binding protein, SmpB, forms part of a ribonucleoprotein complex, provides a template for the resumption of translation elongation, subsequent termination and recycling of stalled ribosomes. In addition, the mRNA-like domain of tmRNA encodes a peptide that tags polypeptides derived from stalled ribosomes for degradation. Streptomyces are unique bacteria that undergo a developmental cycle culminating at sporulation that is at least partly controlled at the level of translation elongation by the abundance of a rare tRNA that decodes UUA codons found in a relatively small number of open reading frames prompting us to examine the role of tmRNA in S. coelicolor. Using a temperature sensitive replicon, we found that the ssrA gene could be disrupted only in cells with an extra-copy wild type gene but not in wild type cells or cells with an extra-copy mutant tmRNA (tmRNADD) encoding a degradation-resistant tag. A cosmid-based gene replacement method that does not include a high temperature step enabled us to disrupt both the ssrA and smpB genes separately and at the same time suggesting that the tmRNA tagging system may be required for cell survival under high temperature. Indeed, mutant cells show growth and sporulation defects at high temperature and under optimal culture conditions. Interestingly, even though these defects can be completely restored by wild type genes, the ΔssrA strain was only partially corrected by tmRNADD. In addition, wildtype tmRNA can restore the hygromycin-resistance to ΔssrA cells while tmRNADD failed to do so suggesting that degradation of aberrant peptides is important for antibiotic resistance. Overall, these results suggest that the tmRNA tagging system plays important roles during Streptomyces growth and sporulation under both normal and stress conditions. PMID:19212432

  17. Regulation of a nickel-cobalt efflux system and nickel homeostasis in a soil actinobacterium Streptomyces coelicolor.

    PubMed

    Kim, Hae Mi; Ahn, Bo-Eun; Lee, Ju-Hyung; Roe, Jung-Hye

    2015-04-01

    In Streptomyces coelicolor, a soil actinobacterium capable of morphological differentiation and complex secondary metabolism, nickel deficiency is sensed by Nur, a Ni-specific Fur family regulator that controls nickel uptake systems (NikABCDE and NikMNOQ) and both Fe-containing and Ni-containing superoxide dismutases (SodF and SodN). On the other hand, the nickel efflux system and its regulator have not been elucidated. In this study, we demonstrate that an ArsR/SmtB family metalloregulator NmtR, a close homologue of NmtR from Mycobacterium tuberculosis, controls a putative efflux pump of P1-type ATPase (NmtA) in S. coelicolor. NmtR binds to the nmtA promoter region to repress its transcription, and is dissociated in the presence of Ni(ii) and Co(ii). Disruption of the nmtA gene makes cells more sensitive to nickel and cobalt, consistent with its predicted role in encoding a Ni-Co-efflux pump. Growth of S. coelicolor in complex YEME medium is only marginally inhibited by up to 0.5 mM Ni(ii), with significant growth retardation at 1 mM. Nur-regulated sodF and nikA genes are repressed at less than 0.1 μM added NiSO4 whereas NmtR-regulated nmtA transcription is induced at 0.5 mM or more Ni(ii). This reveals the extreme sensitivity of S. coelicolor to nickel deficiency as well as tolerance to surplus nickel. How this organism and possibly other actinomycetes have evolved to develop such a highly Ni-tolerant physiology and how the highly sensitive regulator Nur and the obtuse regulator NmtR achieve their characteristic Ni-sensitivity are interesting questions to solve in the future.

  18. Pulse Wave Well Development Demonstration

    SciTech Connect

    Burdick, S.

    2001-02-23

    Conventional methods of well development at the Savannah River Site generate significant volumes of investigative derived waste (IDW) which must be treated and disposed of at a regulated Treatment, Storage, or Disposal (TSD) facility. Pulse Wave technology is a commercial method of well development utilizing bursts of high pressure gas to create strong pressure waves through the well screen zone, extending out into the formation surrounding the well. The patented process is intended to reduce well development time and the amount of IDW generated as well as to micro-fracture the formation to improve well capacity.

  19. Developing dendrites demonstrate unexpected specificity.

    PubMed

    Chalupa, Leo M

    2006-11-22

    Our knowledge of how developing dendrites attain their mature state is still rudimentary. In this issue of Neuron, Mumm et al. rely on time-lapsed analysis of ingrowing dendrites of retinal ganglion cells in transgenic zebrafish to show that this process is much more specific than has been suspected.

  20. New Knowledge from Old: In silico discovery of novel protein domains in Streptomyces coelicolor

    PubMed Central

    Yeats, Corin; Bentley, Stephen; Bateman, Alex

    2003-01-01

    Background Streptomyces coelicolor has long been considered a remarkable bacterium with a complex life-cycle, ubiquitous environmental distribution, linear chromosomes and plasmids, and a huge range of pharmaceutically useful secondary metabolites. Completion of the genome sequence demonstrated that this diversity carried through to the genetic level, with over 7000 genes identified. We sought to expand our understanding of this organism at the molecular level through identification and annotation of novel protein domains. Protein domains are the evolutionary conserved units from which proteins are formed. Results Two automated methods were employed to rapidly generate an optimised set of targets, which were subsequently analysed manually. A final set of 37 domains or structural repeats, represented 204 times in the genome, was developed. Using these families enabled us to correlate items of information from many different resources. Several immediately enhance our understanding both of S. coelicolor and also general bacterial molecular mechanisms, including cell wall biosynthesis regulation and streptomycete telomere maintenance. Discussion Delineation of protein domain families enables detailed analysis of protein function, as well as identification of likely regions or residues of particular interest. Hence this kind of prior approach can increase the rate of discovery in the laboratory. Furthermore we demonstrate that using this type of in silico method it is possible to fairly rapidly generate new biological information from previously uncorrelated data. PMID:12625841

  1. Altered desferrioxamine-mediated iron utilization is a common trait of bald mutants of Streptomyces coelicolor.

    PubMed

    Lambert, Stéphany; Traxler, Matthew F; Craig, Matthias; Maciejewska, Marta; Ongena, Marc; van Wezel, Gilles P; Kolter, Roberto; Rigali, Sébastien

    2014-08-01

    Streptomyces coelicolor is an important model organism for developmental studies of filamentous GC-rich actinobacteria. The genetic characterization of mutants of S. coelicolor blocked at the vegetative mycelium stage, the so-called bald (bld) mutants that are unable to erect spore-forming aerial hyphae, has opened the way to discovering the molecular basis of development in actinomycetes. Desferrioxamine (DFO) production and import of ferrioxamines (FO; iron-complexed DFO) are key to triggering morphogenesis of S. coelicolor and we show here that growth of S. coelicolor on the reference medium for Streptomyces developmental studies is fully dependent on DFO biosynthesis. UPLC-ESI-MS analysis revealed that all bld mutants tested are affected in DFO biosynthesis, with bldA, bldJ, and ptsH mutants severely impaired in DFO production, while bldF, bldK, crr and ptsI mutants overproduce DFO. Morphogenesis of bldK and bldJ mutants was recovered by supplying exogenous iron. Transcript analysis showed that the bldJ mutant is impaired in expression of genes involved in the uptake of FO, whereas transcription of genes involved in both DFO biosynthesis and FO uptake is increased in bldK mutants. Our study allows proposing altered DFO production and/or FO uptake as a novel phenotypic marker of many S. coelicolor bld mutants, and strengthens the role of siderophores and iron acquisition in morphological development of actinomycetes.

  2. Codes & standards research, development & demonstration Roadmap

    SciTech Connect

    None, None

    2008-07-22

    This Roadmap is a guide to the Research, Development & Demonstration activities that will provide data required for SDOs to develop performance-based codes and standards for a commercial hydrogen fueled transportation sector in the U.S.

  3. Developing Noticing of Reasoning through Demonstration Lessons

    ERIC Educational Resources Information Center

    Bragg, Leicha A.; Vale, Colleen

    2014-01-01

    Observation of fellow educators conducting demonstration lessons is one avenue for teachers to develop sensitivity to noticing students' reasoning. We examined teachers' noticing of children's learning behaviours in one demonstration lesson of the "Mathematical Reasoning Professional Learning Research Program" (MRPLRP). The observations…

  4. Demonstration of a System Development Environment

    PubMed Central

    Blum, B. I.

    1985-01-01

    This demonstration illustrates how modern development environments can be used to improve the process of designing and implementing information systems. Following a brief introduction to the topic of application generation, automatic programming, and software environments, one product — TEDIUM* — will be demonstrated.

  5. Development and Demonstration of Ultrafiltration Simulants

    SciTech Connect

    Russell, Renee L.; Billing, Justin M.; Peterson, Reid A.; Rinehart, Donald E.; Smith, Harry D.

    2009-02-24

    According to Bechtel National, Inc. (BNI) Test Specification 24590-PTF-TSP-RT-06-006, Rev 0, Simulant Development to Support the Development and Demonstration of Leaching and Ultrafiltration Pretreatment Processes,” simulants for boehmite, gibbsite, and filtration are to be developed that can be used in subsequent bench and integrated testing of the leaching/filtration processes for the waste treatment plant (WTP). These simulants will then be used to demonstrate the leaching process and to help refine processing conditions which may impact safety basis considerations (Smith 2006). This report documents the results of the filtration simulant development.

  6. Cell immobilization of Streptomyces coelicolor : effect on differentiation and actinorhodin production.

    PubMed

    López-García, María Teresa; Rioseras, Beatriz; Yagüe, Paula; Álvarez, José Ramón; Manteca, Ángel

    2014-06-01

    Streptomycetes are mycelium-forming bacteria that produce two thirds of the clinically relevant secondary metabolites. Despite the fact that secondary metabolite production is activated at specific developmental stages of the Streptomyces spp. life cycle, different streptomycetes show different behaviors, and fermentation conditions need to be optimized for each specific strain and secondary metabolite. Cell-encapsulation constitutes an interesting alternative to classical fermentations, which was demonstrated to be useful in Streptomyces, but development under these conditions remained unexplored. In this work, the influence of cell-encapsulation in hyphae differentiation and actinorhodin production was explored in the model Streptomyces coelicolor strain. Encapsulation led to a delay in growth and to a reduction of mycelium density and cell death. The high proportion of viable hyphae duplicated extracellular actinorhodin production in the encapsulated cultures with respect to the non-encapsulated ones.

  7. Human Support Technology Research, Development and Demonstration

    NASA Technical Reports Server (NTRS)

    Joshi, Jitendra; Trinh, Eugene

    2004-01-01

    The Human Support Technology research, development, and demonstration program address es the following areas at TRL: Advanced Power and Propulsion. Cryogenic fluid management. Closed-loop life support and Habitability. Extravehicular activity systems. Scientific data collection and analysis. and Planetary in-situ resource utilization.

  8. Changes in patterns of ADP-ribosylated proteins during differentiation of Streptomyces coelicolor A3(2) and its development mutants.

    PubMed Central

    Shima, J; Penyige, A; Ochi, K

    1996-01-01

    Mutants resistant to 3-aminobenzamide, a known inhibitor of ADP-ribosyltransferase, were obtained from Streptomyces coelicolor A3(2). One (strain 27) was analyzed in detail. Mutant 27 had a reduced ADP-ribosyl-transferase activity, exhibited substantial changes from the wild type in ADP-ribosylated protein profile during cell aging, and was defective in producing aerial mycelium and antibiotics. A 92-kDa ADP-ribosylated protein disappeared at the onset of differentiation in the parent strain but was present in mutant 27. Four ADP-ribosylated proteins (39, 41, 43, and 46 kDa) appeared at the onset of differentiation in the parent strain but were missing in mutant 27. Failure to ADP-ribosylate these four proteins was detected when the parent strain was grown in the presence of subinhibitory amounts of 3-aminobenzamide. Genetic analysis showed that the mutation, named brgA, conferring resistance to 3-aminobenzamide, cosegregated with the altered phenotypes (i.e., defects in ADP-ribosylation and aerial mycelium formation) and was mapped to a new locus near uraA. The brgA mutants were nonconditionally deficient in producing aerial mycelium and antibiotics, as determined by using various media, and had a morphological and physiological phenotype quite different from that of a bldG mutant carrying a mutation which was previously mapped near uraA. Among the known bld mutants, bldA, bldD, and bldG mutants exhibited a ADP-ribosylated protein profile similar to that of the wild type, while like mutant 27, bldB, bldC, and bldH mutants failed to ADP-ribosylate certain proteins. PMID:8682781

  9. Changes in patterns of ADP-ribosylated proteins during differentiation of Streptomyces coelicolor A3(2) and its development mutants.

    PubMed

    Shima, J; Penyige, A; Ochi, K

    1996-07-01

    Mutants resistant to 3-aminobenzamide, a known inhibitor of ADP-ribosyltransferase, were obtained from Streptomyces coelicolor A3(2). One (strain 27) was analyzed in detail. Mutant 27 had a reduced ADP-ribosyl-transferase activity, exhibited substantial changes from the wild type in ADP-ribosylated protein profile during cell aging, and was defective in producing aerial mycelium and antibiotics. A 92-kDa ADP-ribosylated protein disappeared at the onset of differentiation in the parent strain but was present in mutant 27. Four ADP-ribosylated proteins (39, 41, 43, and 46 kDa) appeared at the onset of differentiation in the parent strain but were missing in mutant 27. Failure to ADP-ribosylate these four proteins was detected when the parent strain was grown in the presence of subinhibitory amounts of 3-aminobenzamide. Genetic analysis showed that the mutation, named brgA, conferring resistance to 3-aminobenzamide, cosegregated with the altered phenotypes (i.e., defects in ADP-ribosylation and aerial mycelium formation) and was mapped to a new locus near uraA. The brgA mutants were nonconditionally deficient in producing aerial mycelium and antibiotics, as determined by using various media, and had a morphological and physiological phenotype quite different from that of a bldG mutant carrying a mutation which was previously mapped near uraA. Among the known bld mutants, bldA, bldD, and bldG mutants exhibited a ADP-ribosylated protein profile similar to that of the wild type, while like mutant 27, bldB, bldC, and bldH mutants failed to ADP-ribosylate certain proteins.

  10. Vibration isolation technology development to demonstration

    NASA Technical Reports Server (NTRS)

    Grodsinsky, Carlos

    1992-01-01

    The main thrust of these studies has resulted in an active inertial feedforward/feedback isolation system. This prototype magnetic suspension system has been demonstrated in a laboratory setting in six degrees-of-freedom and has been preliminarily characterized in its isolation performance with favorable results. This isolation system consists of a closed loop digital control system referencing a platform around six relative and six inertial sensors. These sensors control the isolated mass through nine attractive electromagnetic actuators with a system capability of +/- three-tenths of an inch travel in three dimensions. The development of a prototype system from design to fabrication leads directly into the demonstration phase of the project which will attempt a low gravity environmental demonstration of engineering hardware for the isolation of a scientific payload. The demonstration phase of the project will use an aircraft low gravity maneuver to establish a research testbed for the study of isolation hardware and control strategies in an off-loaded environment. In developing this demonstration capability the Lewis Learjet aircraft has been characterized through its parabolic flight maneuvers and a trunnioned experimental volume has been designed for the test of both active and passive isolation packages. This vibration isolation testbed is operational and has two data acquisition systems available for both autonomous and interactive operation, with a combined input capability of 32 channels.

  11. Integrated powerhead demonstration full flow cycle development

    NASA Astrophysics Data System (ADS)

    Jones, J. Mathew; Nichols, James T.; Sack, William F.; Boyce, William D.; Hayes, William A.

    1998-01-01

    The Integrated Powerhead Demonstration (IPD) is a 1,112,000 N (250,000 lbf) thrust (at sea level) LOX/LH2 demonstration of a full flow cycle in an integrated system configuration. Aerojet and Rocketdyne are on contract to the Air Force Research Laboratory to design, develop, and deliver the required components, and to provide test support to accomplish the demonstration. Rocketdyne is on contract to provide a fuel and oxygen turbopump, a gas-gas injector, and system engineering and integration. Aerojet is on contract to provide a fuel and oxygen preburner, a main combustion chamber, and a nozzle. The IPD components are being designed with Military Spaceplane (MSP) performance and operability requirements in mind. These requirements include: lifetime >=200 missions, mean time between overhauls >=100 cycles, and a capability to throttle from 20% to 100% of full power. These requirements bring new challenges both in designing and testing the components. This paper will provide some insight into these issues. Lessons learned from operating and supporting the space shuttle main engine (SSME) have been reviewed and incorporated where applicable. The IPD program will demonstrate phase I goals of the Integrated High Payoff Rocket Propulsion Technology (IHPRPT) program while demonstrating key propulsion technologies that will be available for MSP concepts. The demonstration will take place on Test Stand 2A at the Air Force Research Laboratory at Edwards AFB. The component tests will begin in 1999 and the integrated system tests will be completed in 2002.

  12. Servicer system demonstration plan and capability development

    NASA Technical Reports Server (NTRS)

    1987-01-01

    An orbital maneuvering vehicle (OMV) front end kit is defined which is capable of performing in-situ fluid resupply and modular maintenance of free flying spacecraft based on the integrated orbital servicing system (IOSS) concept. The compatibility of the IOSS to perform gas and fluid umbilical connect and disconnect functions utilizing connect systems currently available or in development is addressed. A series of tasks involving on-orbit servicing and the engineering test unit (ETU) of the on-orbit service were studied. The objective is the advancement of orbital servicing by expanding the Spacecraft Servicing Demonstration Plan (SSDP) to include detail demonstration planning using the Multimission Modular Spacecraft (MMS) and upgrading the ETU control.

  13. Fuel consolidation demonstration: Consolidation concept development

    SciTech Connect

    Not Available

    1990-02-01

    EPRI, Northeast utilities Service Company (NUSCO), DOE, Baltimore Gas Electric Company, and Combustion Engineering, Inc. (C-E) are engaged in a program to develop a system for consolidating spent fuel, in which the consolidated fuel will be licensable by NRC for storage in the spent-fuel storage pool. Fuel consolidation offers a means of substantially increasing the capacity of spent-fuel storage pools. Consolidation equipment design, development, construction, and testing are being performed by C-E in Windsor, Connecticut. Seismic and structural evaluation of the capability of the Millstone Unit 2 spent-fuel pool and building to accommodate the increased fuel capacity is being conducted by NUSCO. NUSCO plans to obtain a license to store consolidated fuel in the Millstone-2 spent-fuel storage pool. NUSCO also plans to perform a hot demonstration of the integrated consolidation system with spent fuel at Millstone-2. This report describes the consolidation system design that forms the basis for the detailed design of the equipment comprising the system, including information on the fabrication and testing of the equipment. Appendix B describes an evaluation of the ability of the system under development to consolidate LWR spent-fuel assemblies other than the 14 {times} 14 fuel of C-E design stored at Millstone-2. A comparison was made of fuel-assembly designs on the basis of information available in open literature. It was concluded that with appropriate dimensional modifications the spent-fuel consolidation system equipment design is applicable to almost all PWR fuel-assembly configurations. 8 refs., 20 figs.

  14. Multiyear Research, Development and Demonstration Plan

    SciTech Connect

    EERE

    2012-03-16

    The Fuel Cell Technologies Office Multi-Year Research, Development, and Demonstration (MYRD&D) Plan describes the goals, objectives, technical targets, tasks, and schedules for all activities within the Fuel Cell Technologies (FCT) Office, which is part of the U.S. Department of Energy's Office of Energy Efficiency and Renewable Energy. The MYRD&D Plan is a living document, which is revised periodically to reflect progress in the technologies, revisions to developmental timelines and targets, updates based on external reviews, and changes in the scope of the FCT Office. The document was first published in 2003, and revised in 2005. The document was significantly revised in 2012 to reflect scientific advancements and the changing technological landscape. Any revisions made to the MYRD&D Plan are conducted through a rigorous Change Control process as documented in the Systems Integration section of this report. The hydrogen and fuel cell activities within DOE continue to receive extensive review by stakeholders in the hydrogen and fuel cell community, including panels of the National Research Council and the National Academy of Engineering.

  15. Servicers system demonstration plan and capability development

    NASA Technical Reports Server (NTRS)

    Bulboaca, M. A.; Cuseo, J. A.; Derocher, W. L., Jr.; Maples, R. W.; Reynolds, P. C.; Sterrett, R. A.

    1985-01-01

    A plan for the demonstration of the exchange of Multi-Mission Modular Spacecraft (MMS) modules using the servicer mechanism Engineering Test Unit (ETU) was prepared and executed. The plan included: establishment of requirements, conceptual design, selection of MMS spacecraft mockup configuration, selection of MMS module mockup configuration, evaluation of adequacy of ETU load capability, and selection of a stowage rack arrangement. The MMS module exchange demonstration mockup equipment was designed, fabricated, checked out, shipped, installed, and demonstrated.

  16. The Absence of Pupylation (Prokaryotic Ubiquitin-Like Protein Modification) Affects Morphological and Physiological Differentiation in Streptomyces coelicolor

    PubMed Central

    Seghezzi, Nicolas; Duchateau, Magalie; Gominet, Myriam; Kofroňová, Olga; Benada, Oldřich; Mazodier, Philippe

    2015-01-01

    ABSTRACT Protein turnover is essential in all living organisms for the maintenance of normal cell physiology. In eukaryotes, most cellular protein turnover involves the ubiquitin-proteasome pathway, in which proteins tagged with ubiquitin are targeted to the proteasome for degradation. In contrast, most bacteria lack a proteasome but harbor proteases for protein turnover. However, some actinobacteria, such as mycobacteria, possess a proteasome in addition to these proteases. A prokaryotic ubiquitination-like tagging process in mycobacteria was described and was named pupylation: proteins are tagged with Pup (prokaryotic ubiquitin-like protein) and directed to the proteasome for degradation. We report pupylation in another actinobacterium, Streptomyces coelicolor. Both the morphology and life cycle of Streptomyces species are complex (formation of a substrate and aerial mycelium followed by sporulation), and these bacteria are prolific producers of secondary metabolites with important medicinal and agricultural applications. The genes encoding the pupylation system in S. coelicolor are expressed at various stages of development. We demonstrated that pupylation targets numerous proteins and identified 20 of them. Furthermore, we established that abolition of pupylation has substantial effects on morphological and metabolic differentiation and on resistance to oxidative stress. In contrast, in most cases, a proteasome-deficient mutant showed only modest perturbations under the same conditions. Thus, the phenotype of the pup mutant does not appear to be due solely to defective proteasomal degradation. Presumably, pupylation has roles in addition to directing proteins to the proteasome. IMPORTANCE Streptomyces spp. are filamentous and sporulating actinobacteria, remarkable for their morphological and metabolic differentiation. They produce numerous bioactive compounds, including antifungal, antibiotic, and antitumor compounds. There is therefore considerable interest in

  17. SCO5745, a Bifunctional RNase J Ortholog, Affects Antibiotic Production in Streptomyces coelicolor

    PubMed Central

    Bralley, Patricia; Aseem, Madiha

    2014-01-01

    The bacterial RNases J are considered bifunctional RNases possessing both endo- and exonucleolytic activities. We have isolated an RNase J ortholog from Streptomyces coelicolor encoded by the gene sco5745. We overexpressed a decahistidine-tagged version of SCO5745 and purified the overexpressed protein by immobilized metal ion affinity chromatography. We demonstrated the presence of both 5′-to-3′ exonucleolytic and endonucleolytic activities on the Bacillus subtilis thrS transcript. Exonucleoytic activity predominated with 5′ monophosphorylated thrS, while endonucleolytic activity predominated with 5′ triphosphorylated thrS. While sco5745 is the only RNase J allele in S. coelicolor, the gene is not essential. Its disruption resulted in delayed production of the antibiotic actinorhodin, overproduction of undecylprodigiosin, and diminished production of the calcium-dependent antibiotic, in comparison with the parental strain. PMID:24415725

  18. Game Changing Development Demonstration Video 2014

    NASA Video Gallery

    The Game Changing Development Program is a part of NASA’s Space Technology Mission Directorate. The Program advances space technologies that may lead to entirely new approaches for the Agency’s fut...

  19. Geothermal Energy Research Development and Demonstration Program

    SciTech Connect

    Not Available

    1980-06-01

    The Federal program's goal, strategy, plans, and achievements are summarized. In addition, geothermal development by state and local governments and, where available, by the private sector is described. (MHR)

  20. Demonstration and Validation Assets: User Manual Development

    SciTech Connect

    2008-06-30

    This report documents the development of a database-supported user manual for DEMVAL assets in the NSTI area of operations and focuses on providing comprehensive user information on DEMVAL assets serving businesses with national security technology applications in southern New Mexico. The DEMVAL asset program is being developed as part of the NSPP, funded by both Department of Energy (DOE) and NNSA. This report describes the development of a comprehensive user manual system for delivering indexed DEMVAL asset information to be used in marketing and visibility materials and to NSTI clients, prospective clients, stakeholders, and any person or organization seeking it. The data about area DEMVAL asset providers are organized in an SQL database with updateable application structure that optimizes ease of access and customizes search ability for the user.

  1. Nursing Professional Development Organizational Value Demonstration Project.

    PubMed

    Harper, Mary G; Aucoin, Julia; Warren, Joan I

    2016-01-01

    A common question nursing professional development (NPD) practitioners ask is, "How many NPD practitioners should my organization have?" This study examined correlations among facility size and structure, NPD practitioner characteristics and time in service, and organizational outcomes. Organizations with a higher rate of NPD full-time equivalents per bed had higher patient satisfaction with nurses' communication and provision of discharge instruction on their HCAHPS (Hospital Consumer Assessment of Healthcare Provider and Systems) scores.

  2. Production of specialized metabolites by Streptomyces coelicolor A3(2).

    PubMed

    van Keulen, Geertje; Dyson, Paul J

    2014-01-01

    The actinomycetes are well-known bioactive natural product producers, comprising the Streptomycetes, the richest drug-prolific family in all kingdoms, producing therapeutic compounds for the areas of infection, cancer, circulation, and immunity. Completion and annotation of many actinomycete genomes has highlighted further how proficient these bacteria are in specialized metabolism, which have been largely underexploited in traditional screening programs. The genome sequence of the model strain Streptomyces coelicolor A3(2), and subsequent development of genomics-driven approaches to understand its large specialized metabolome, has been key in unlocking the high potential of specialized metabolites for natural product genomics-based drug discovery. This review discusses systematically the biochemistry and genetics of each of the specialized metabolites of S. coelicolor and describes metabolite transport processes for excretion and complex regulatory patterns controlling biosynthesis.

  3. Integrated Metabolomics Approach Facilitates Discovery of an Unpredicted Natural Product Suite from Streptomyces coelicolor M145

    PubMed Central

    Sidebottom, Ashley M.; Johnson, Andrew R.; Karty, Jonathan A.; Trader, Darci J.; Carlson, Erin E.

    2013-01-01

    Natural products exhibit a broad range of biological properties and have been a crucial source of therapeutic agents and novel scaffolds. Although bacterial secondary metabolomes are widely explored, they remain incompletely cataloged by current isolation and characterization strategies. To identify metabolites residing in unexplored chemical space, we have developed an integrated discovery approach that combines bacterial growth perturbation, accurate mass spectrometry, comparative mass spectra data analysis, and fragmentation spectra clustering for the identification of low-abundant, novel compounds from complex biological matrices. In this investigation, we analyzed the secreted metabolome of the extensively studied Actinomycete, Streptomyces coelicolor M145, and discovered a low-abundant suite of 15 tri-hydroxamate, amphiphilic siderophores. Compounds in this class have primarily been observed in marine microorganisms making their detection in the soil-dwelling S. coelicolor M145 significant. At least ten of these ferrioxamine-based molecules are not known to be produced by any organism and none have previously been detected from S. coelicolor M145. In addition, we confirmed the production of ferrioxamine D1, a relatively hydrophilic family member that has not been shown to be biosynthesized by this organism. The identified molecules are part of only a small list of secondary metabolites that have been discovered since sequencing of S. coelicolor M145 revealed that it possessed numerous putative secondary metabolite-producing gene clusters with no known metabolites. Thus, the identified siderophores represent the unexplored metabolic potential of both well-studied and new organisms that could be uncovered with our sensitive and robust approach. PMID:23777274

  4. Repression of Antibiotic Production and Sporulation in Streptomyces coelicolor by Overexpression of a TetR Family Transcriptional Regulator ▿ †

    PubMed Central

    Xu, Delin; Seghezzi, Nicolas; Esnault, Catherine; Virolle, Marie-Joelle

    2010-01-01

    The overexpression of a regulatory gene of the TetR family (SCO3201) originating either from Streptomyces lividans or from Streptomyces coelicolor was shown to strongly repress antibiotic production (calcium-dependent antibiotic [CDA], undecylprodigiosin [RED], and actinorhodin [ACT]) of S. coelicolor and of the ppk mutant strain of S. lividans. Curiously, the overexpression of this gene also had a strong inhibitory effect on the sporulation process of S. coelicolor but not on that of S. lividans. SCO3201 was shown to negatively regulate its own transcription, and its DNA binding motif was found to overlap its −35 promoter sequence. The interruption of this gene in S. lividans or S. coelicolor did not lead to any obvious phenotypes, indicating that when overexpressed SCO3201 likely controls the expression of target genes of other TetR regulators involved in the regulation of the metabolic and morphological differentiation process in S. coelicolor. The direct and functional interaction of SCO3201 with the promoter region of scbA, a gene under the positive control of the TetR-like regulator, ScbR, was indeed demonstrated by in vitro as well as in vivo approaches. PMID:20935121

  5. 10 CFR 1021.212 - Research, development, demonstration, and testing.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 10 Energy 4 2010-01-01 2010-01-01 false Research, development, demonstration, and testing. 1021... ACT IMPLEMENTING PROCEDURES DOE Decisionmaking § 1021.212 Research, development, demonstration, and..., development, demonstration, and testing for new technologies (40 CFR 1502.4(c)(3)). Adoption of such...

  6. Multiple biosynthetic and uptake systems mediate siderophore-dependent iron acquisition in Streptomyces coelicolor A3(2) and Streptomyces ambofaciens ATCC 23877.

    PubMed

    Barona-Gómez, Francisco; Lautru, Sylvie; Francou, Francois-Xavier; Leblond, Pierre; Pernodet, Jean-Luc; Challis, Gregory L

    2006-11-01

    Siderophore-mediated iron acquisition has been well studied in many bacterial pathogens because it contributes to virulence. In contrast, siderophore-mediated iron acquisition by saprophytic bacteria has received relatively little attention. The independent identification of the des and cch gene clusters that direct production of the tris-hydroxamate ferric iron-chelators desferrioxamine E and coelichelin, respectively, which could potentially act as siderophores in the saprophyte Streptomyces coelicolor A3(2), has recently been reported. Here it is shown that the des cluster also directs production of desferrioxamine B in S. coelicolor and that very similar des and cch clusters direct production of desferrioxamines E and B, and coelichelin, respectively, in Streptomyces ambofaciens ATCC 23877. Sequence analyses of the des and cch clusters suggest that components of ferric-siderophore uptake systems are also encoded within each cluster. The construction and analysis of a series of mutants of S. coelicolor lacking just biosynthetic genes or both the biosynthetic and siderophore uptake genes from the des and cch clusters demonstrated that coelichelin and desferrioxamines E and B all function as siderophores in this organism and that at least one of these metabolites is required for growth under defined conditions even in the presence of significant quantities of ferric iron. These experiments also demonstrated that a third siderophore uptake system must be present in S. coelicolor, in addition to the two encoded within the cch and des clusters, which show selectivity for coelichelin and desferrioxamine E, respectively. The ability of the S. coelicolor mutants to utilize a range of exogenous xenosiderophores for iron acquisition was also examined, showing that the third siderophore-iron transport system has broad specificity for tris-hydroxamate-containing siderophores. Together, these results define a complex system of multiple biosynthetic and uptake pathways for

  7. 40 CFR 258.4 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... demonstration permits. 258.4 Section 258.4 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... State may issue a research, development, and demonstration permit for a new MSWLF unit, existing MSWLF...). (b) The Director of an approved State may issue a research, development, and demonstration permit...

  8. 40 CFR 258.4 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... demonstration permits. 258.4 Section 258.4 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... State may issue a research, development, and demonstration permit for a new MSWLF unit, existing MSWLF...). (b) The Director of an approved State may issue a research, development, and demonstration permit...

  9. 40 CFR 258.4 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... demonstration permits. 258.4 Section 258.4 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... State may issue a research, development, and demonstration permit for a new MSWLF unit, existing MSWLF...). (b) The Director of an approved State may issue a research, development, and demonstration permit...

  10. Spatio-temporal expression of the pathway-specific regulatory gene redD in S. coelicolor.

    PubMed

    Zhou, Li-hua; Li, Yu-qin; Li, Yong-quan; Wu, Dan

    2005-06-01

    Confocal laser scanning microscopy was used to observe the spatio-temporal expression of the pathway-specific gene redD during S. coelicolor cell cultivation. The corresponding mutant S. coelicolor lyqRY1522 carrying redD::eyfp in the chromosome was constructed. The temporal expression results of the fusion protein during submerged cultivation demonstrated that expression of redD began in the transition phase, continuing through the exponential growth phase to the stationary phase, and reached maximum in the stationary phase. On the other hand, redD was expressed only in substrate mycelia during solid-state culture, while aerial mycelia remained essentially non-fluorescent throughout culture. Results demonstrated that the expression pattern of redD coincides with that of the biosynthesis of the antibiotics during culture, revealing a direct correlation between the spatio-temporal distribution of regulatory gene expression and second metabolism.

  11. Development of a Defence Distributed Computing Environment (DCE) Database Demonstrator,

    DTIC Science & Technology

    1995-11-01

    This report discusses the development of a Defence Distributed Computing Environment (DCE) database demonstrator program. The Demonstrator program...showcases the interoperability, portability, survivability and security features of Open Software Foundation’s Distributed Computing Environment.

  12. Alternative-fueled truck demonstration natural gas program: Caterpillar G3406LE development and demonstration

    SciTech Connect

    1995-06-01

    In 1990, the California Energy Commission, the South Coast Air Quality Management District, and the Southern California Gas Company joined together to sponsor the development and demonstration of compressed natural gas engines for Class 8 heavy-duty line-haul trucking applications. This program became part of an overall Alternative-Fueled Truck Demonstration Program, with the goal of advancing the technological development of alternative-fueled engines. The demonstration showed natural gas to be a technically viable fuel for Class 8 truck engines.

  13. 40 CFR 270.65 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 26 2010-07-01 2010-07-01 false Research, development, and... Special Forms of Permits § 270.65 Research, development, and demonstration permits. (a) The Administrator may issue a research, development, and demonstration permit for any hazardous waste treatment...

  14. 40 CFR 270.65 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 28 2012-07-01 2012-07-01 false Research, development, and... Special Forms of Permits § 270.65 Research, development, and demonstration permits. (a) The Administrator may issue a research, development, and demonstration permit for any hazardous waste treatment...

  15. Development of a laboratory demonstration model active cleaning device

    NASA Technical Reports Server (NTRS)

    Shannon, R. L.; Gillette, R. B.

    1975-01-01

    A laboratory demonstration model of a device for removing contaminant films from optical surfaces in space was developed. The development of a plasma tube, which would produce the desired cleaning effects under high vacuum conditions, represented the major problem in the program. This plasma tube development is discussed, and the resulting laboratory demonstration-model device is described.

  16. 17 CFR 256.188 - Research, development, or demonstration expenditures.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... demonstration expenditures. 256.188 Section 256.188 Commodity and Securities Exchanges SECURITIES AND EXCHANGE... demonstration expenditures. (a) The service company shall utilize this account, where appropriate, for the cost of all expenditures for research, development or demonstration undertaken by or sponsored through...

  17. 10 CFR 1021.212 - Research, development, demonstration, and testing.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 10 Energy 4 2013-01-01 2013-01-01 false Research, development, demonstration, and testing. 1021... ACT IMPLEMENTING PROCEDURES DOE Decisionmaking § 1021.212 Research, development, demonstration, and testing. (a) This section applies to the adoption and application of programs that involve...

  18. 10 CFR 1021.212 - Research, development, demonstration, and testing.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 10 Energy 4 2014-01-01 2014-01-01 false Research, development, demonstration, and testing. 1021... testing. (a) This section applies to the adoption and application of programs that involve research, development, demonstration, and testing for new technologies (40 CFR 1502.4(c)(3)). Adoption of such...

  19. The Development and Demonstration of The Metric Assessment Tool

    DTIC Science & Technology

    1993-09-01

    APIT/GCM/LAS/93S-3 A04 Ai 7.5 4 . " . THE DEVELOPMENT AND DEMONSTRATION OF THE METRIC ASSESSMENT TOOL THESIS Cynthia A. Campbell, GS-13, USAF and...Av•JI:a)iity Ccdes Avaii "•tc, or Dist 6pi-claI * iiV AFIT/GCM/LAS/93S-3 THE DEVELOPMENT AND DEMONSTRATION OF THE METRIC ASSESSMENT TOOL THESIS...thesis was to develop and demonstrate a Metric Assessment Tool for the purpose of evaluating and improving process measurements. We narrowed our study

  20. Codes and standards research, development and demonstration roadmap

    SciTech Connect

    None, None

    2008-07-22

    C&S RD&D Roadmap - 2008: This Roadmap is a guide to the Research, Development & Demonstration activities that will provide data required for Standards Development Organizations (SDOs) to develop performance-based codes and standards for a commercial hydrogen fueled transportation sector in the U.S.

  1. 10 CFR 1021.212 - Research, development, demonstration, and testing.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ..., development, demonstration, and testing for new technologies (40 CFR 1502.4(c)(3)). Adoption of such programs... (if otherwise required by this part) as soon as environmental effects can be meaningfully...

  2. 10 CFR 1021.212 - Research, development, demonstration, and testing.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ..., development, demonstration, and testing for new technologies (40 CFR 1502.4(c)(3)). Adoption of such programs... (if otherwise required by this part) as soon as environmental effects can be meaningfully...

  3. Experiments with Corn To Demonstrate Plant Growth and Development.

    ERIC Educational Resources Information Center

    Haldeman, Janice H.; Gray, Margarit S.

    2000-01-01

    Explores using corn seeds to demonstrate plant growth and development. This experiment allows students to formulate hypotheses, observe and record information, and practice mathematics. Presents background information, materials, procedures, and observations. (SAH)

  4. Novel Two-Component Systems Implied in Antibiotic Production in Streptomyces coelicolor

    PubMed Central

    Yepes, Ana; Rico, Sergio; Rodríguez-García, Antonio; Santamaría, Ramón I.; Díaz, Margarita

    2011-01-01

    The abundance of two-component systems (TCSs) in Streptomyces coelicolor A3(2) genome indicates their importance in the physiology of this soil bacteria. Currently, several TCSs have been related to antibiotic regulation, and the purpose in this study was the characterization of five TCSs, selected by sequence homology with the well-known absA1A2 system, that could also be associated with this important process. Null mutants of the five TCSs were obtained and two mutants (ΔSCO1744/1745 and ΔSCO4596/4597/4598) showed significant differences in both antibiotic production and morphological differentiation, and have been renamed as abr (antibiotic regulator). No detectable changes in antibiotic production were found in the mutants in the systems that include the ORFs SCO3638/3639, SCO3640/3641 and SCO2165/2166 in any of the culture conditions assayed. The system SCO1744/1745 (AbrA1/A2) was involved in negative regulation of antibiotic production, and acted also as a negative regulator of the morphological differentiation. By contrast, the system SCO4596/4597/4598 (AbrC1/C2/C3), composed of two histidine kinases and one response regulator, had positive effects on both morphological development and antibiotic production. Microarray analyses of the ΔabrC1/C2/C3 and wild-type transcriptomes revealed downregulation of actII-ORF4 and cdaR genes, the actinorhodin and calcium-dependent antibiotic pathway-specific regulators respectively. These results demonstrated the involvement of these new two-component systems in antibiotic production and morphological differentiation by different approaches. One is a pleiotropic negative regulator: abrA1/A2. The other one is a positive regulator composed of three elements, two histidine kinases and one response regulator: abrC1/C2/C3. PMID:21625497

  5. Synthesis and uptake of the compatible solutes ectoine and 5-hydroxyectoine by Streptomyces coelicolor A3(2) in response to salt and heat stresses.

    PubMed

    Bursy, Jan; Kuhlmann, Anne U; Pittelkow, Marco; Hartmann, Holger; Jebbar, Mohamed; Pierik, Antonio J; Bremer, Erhard

    2008-12-01

    Streptomyces coelicolor A3(2) synthesizes ectoine and 5-hydroxyectoine upon the imposition of either salt (0.5 M NaCl) or heat stress (39 degrees C). The cells produced the highest cellular levels of these compatible solutes when both stress conditions were simultaneously imposed. Protection against either severe salt (1.2 M NaCl) or heat stress (39 degrees C) or a combination of both environmental cues could be accomplished by adding low concentrations (1 mM) of either ectoine or 5-hydroxyectoine to S. coelicolor A3(2) cultures. The best salt and heat stress protection was observed when a mixture of ectoine and 5-hydroxyectoine (0.5 mM each) was provided to the growth medium. Transport assays with radiolabeled ectoine demonstrated that uptake was triggered by either salt or heat stress. The most effective transport and accumulation of [(14)C]ectoine by S. coelicolor A3(2) were achieved when both environmental cues were simultaneously applied. Our results demonstrate that the accumulation of the compatible solutes ectoine and 5-hydroxyectoine allows S. coelicolor A3(2) to fend off the detrimental effects of both high salinity and high temperature on cell physiology. We also characterized the enzyme (EctD) required for the synthesis of 5-hydroxyectoine from ectoine, a hydroxylase of the superfamily of the non-heme-containing iron(II)- and 2-oxoglutarate-dependent dioxygenases (EC 1.14.11). The gene cluster (ectABCD) encoding the enzymes for ectoine and 5-hydroxyectoine biosynthesis can be found in the genome of S. coelicolor A3(2), Streptomyces avermitilis, Streptomyces griseus, Streptomyces scabiei, and Streptomyces chrysomallus, suggesting that these compatible solutes play an important role as stress protectants in the genus Streptomyces.

  6. Effects of simulated microgravity and spaceflight on morphological differentiation and secondary metabolism of Streptomyces coelicolor A3(2).

    PubMed

    Huang, Bing; Liu, Ning; Rong, Xiaoying; Ruan, Jisheng; Huang, Ying

    2015-05-01

    As well-known antibiotic-producing and filamentous bacteria, streptomycetes can be an ideal model to study the effects of microgravity on microbial development and antibiotic production. In this study, the model organism Streptomyces coelicolor A3(2) was exposed to simulated microgravity (SMG) on a rotating clinostat and microgravity (μg) on the Shenzhou-8 spacecraft. The strain exhibited some similar responses under both conditions. Compared with the controls, its life cycle in agar medium was shortened relatively, and the sporulation process was accelerated with higher accumulation of the gray spore pigment; the liquid cultures yielded more cell biomass, coupled with thicker, more fragmented, and well-dispersed hyphae of the μg spaceflight samples. Global transcriptional analysis verified that most of the differentially expressed genes involved in morphological differentiation of S. coelicolor were upregulated during days 4-6 under SMG conditions, notably the whi genes (whiD, sigF, and whiE). Production of actinorhodin (ACT) in agar cultures decreased under both conditions while undecylprodigiosin (RED) was produced earlier, which were consistent with the transcriptional levels of act and red gene clusters. Meanwhile, expression of the gene clusters for calcium-dependent antibiotic (CDA), methylenomycin (MMY), and a cryptic polyketide (CPK) was unchanged, downregulated, and upregulated, respectively, the latter of which might contribute to the enhanced activity of S. coelicolor against Bacillus subtilis under microgravity. Our study provides new insights into the morphological and secondary metabolic responses of streptomycetes to microgravity.

  7. Developmentally regulated cleavage of tRNAs in the bacterium Streptomyces coelicolor

    PubMed Central

    Haiser, Henry J.; Karginov, Fedor V.; Hannon, Gregory J.; Elliot, Marie A.

    2008-01-01

    The ability to sense and respond to environmental and physiological signals is critical for the survival of the soil-dwelling Gram-positive bacterium Streptomyces coelicolor. Nutrient deprivation triggers the onset of a complex morphological differentiation process that involves the raising of aerial hyphae and formation of spore chains, and coincides with the production of a diverse array of clinically relevant antibiotics and other secondary metabolites. These processes are tightly regulated; however, the genes and signals involved have not been fully elucidated. Here, we report a novel tRNA cleavage event that follows the same temporal regulation as morphological and physiological differentiation, and is growth medium dependent. All tRNAs appear to be susceptible to cleavage; however, there appears to be a bias towards increased cleavage of those tRNAs that specify highly utilized codons. In contrast to what has been observed in eukaryotes, accumulation of tRNA halves in S. coelicolor is not significantly affected by amino acid starvation, and is also not affected by induction of the stringent response or inhibition of ribosome function. Mutants defective in aerial development and antibiotic production exhibit altered tRNA cleavage profiles relative to wild-type strains. PMID:18084030

  8. Cytochrome P450 107U1 is required for sporulation and antibiotic production in Streptomyces coelicolor

    PubMed Central

    Tian, Zhenghua; Cheng, Qian; Yoshimoto, Francis K.; Lei, Li; Lamb, David C.; Guengerich, F. Peter

    2013-01-01

    The filamentous bacterium Streptomyces coelicolor has a complex life cycle involving the formation of hair-like aerial mycelia on the colony surface, which differentiate into chains of spores. Genes required for the initiation of aerial mycelium formation have been termed ‘bld’ (bald), describing the smooth, undifferentiated colonies of mutant strains. We report the identification of a new bld gene designated as sco3099 and biochemical analysis of its encoded enzyme, cytochrome P450 (P450, or CYP) 107U1. Deletion of sco3099 resulted in a mutant defective in aerial hyphae sporulation and sensitive to heat shock, indicating that P450 107U1 plays a key role in growth and development of S. coelicolor. This is the first P450 reported to participate in a sporulation process in Streptomycetes. The substrate and catalytic properties of P450 107U1 were further investigated in mass spectrometry-based metabolomic studies. Glycocholic acid (from the medium) was identified as a substrate of P450 107U1 and was oxidized to glyco-7-oxo-deoxycholic acid. Although this reaction is apparently not relevant to the observed sporulation deficiency, it suggests that P450 107U1 might exert its physiological function by oxidizing other steroid-like molecules. PMID:23357279

  9. Aging aircraft NDI Development and Demonstration Center (AANC): An overview

    NASA Astrophysics Data System (ADS)

    Walter, Patrick L.

    1992-07-01

    A major center with emphasis on validation of nondestructive inspection (NDI) techniques for aging aircraft, the Aging Aircraft NDI Development and Demonstration Center (AANC), has been funded by the FAA at Sandia National Laboratories. The Center has been assigned specific tasks in developing techniques for the nondestructive inspection of static engine parts, assessing inspection reliability (POD experiments), developing testbeds for NDI validation, maintaining a FAA library of characterized aircraft structural test specimens, and leasing a hangar to house a high flight cycle transport aircraft for use as a full scale test bed.

  10. Aging Aircraft NDI Development and Demonstration Center (AANC): An overview

    SciTech Connect

    Walter, P.L.

    1991-01-01

    A major center with emphasis on validation of nondestructive inspection techniques for aging aircraft, the Aging Aircraft NDI Development and Demonstration Center (AANC), has been funded by the FAA at Sandia National Laboratories. The Center has been assigned specific tasks in developing techniques for the nondestructive inspection of static engine parts, assessing inspection reliability (POD experiments), developing test beds for nondestructive inspection validation, maintaining a FAA library of characterized aircraft structural test specimens, and leasing a hangar to house a high flight cycle transport aircraft for use as a full scale test bed. 3 refs.

  11. Satellite Communications for Aeronautics Applications: Technology Development and Demonstration

    NASA Technical Reports Server (NTRS)

    Kerczewski, Robert J.; Hoder, Douglas J.; Zakrajsek, Robert J.

    2001-01-01

    The National Aeronautics and Space Administration (NASA) is performing research and development to improve the safety and increase the capacity of the National Airspace System (NAS). Improved communications, especially to and from the aircraft flight deck, has been identified as an essential enabling technology for future improvements to the air traffic management system and aviation safety. NASA's Glenn Research Center is engaged in research and development of satellite communications technologies for aeronautical applications. A mobile aero terminal has been developed for use with Ku band commercial communications satellites. This experimental terminal will be used in mobile ground and air-based tests and demonstrations during 2000-2004. This paper will describe the basic operational parameters of the Ku Band aero terminal, the communications architecture it is intended to demonstrate, and the key technology issues being addressed in the tests and demonstrations. The design of the Ku Band aero terminal and associated ground testbed, planned tests and demonstrations, and results to date will be presented.

  12. Technology Development and Demonstration Concepts for the Space Elevator

    NASA Technical Reports Server (NTRS)

    Smitherman, David V., Jr.

    2004-01-01

    During the 1990s several discoveries and advances in the development of carbon nano-tube (CNT) materials indicated that material strengths many times greater than common high-strength composite materials might be possible. Progress in the development of this material led to renewed interest in the space elevator concept for construction of a tether structure from the surface of the Earth through a geostationary orbit (GEO) and thus creating a new approach to Earth-to-orbit transportation infrastructures. To investigate this possibility the author, in 1999, managed for NASA a space elevator work:hop at the Marshall Space Flight Center to explore the potential feasibility of space elevators in the 21 century, and to identify the critical technologies and demonstration missions needed to make development of space elevators feasible. Since that time, a NASA Institute for Advanced Concepts (NIAC) funded study of the Space Elevator proposed a concept for a simpler first space elevator system using more near-term technologies. This paper will review some of the latest ideas for space elevator development, the critical technologies required, and some of the ideas proposed for demonstrating the feasibility for full-scale development of an Earth to GEO space elevator. Critical technologies include CNT composite materials, wireless power transmission, orbital object avoidance, and large-scale tether deployment and control systems. Numerous paths for technology demonstrations have been proposed utilizing ground experiments, air structures. LEO missions, the space shuttle, the international Space Station, GEO demonstration missions, demonstrations at the lunar L1 or L2 points, and other locations. In conclusion, this paper finds that the most critical technologies for an Earth to GEO space elevator include CNT composite materials development and object avoidance technologies; that lack of successful development of these technologies need not preclude continued development of

  13. [Optimized sample preparation for metabolome studies on Streptomyces coelicolor].

    PubMed

    Li, Yihong; Li, Shanshan; Ai, Guomin; Wang, Weishan; Zhang, Buchang; Yang, Keqian

    2014-04-01

    Streptomycetes produce many antibiotics and are important model microorgansims for scientific research and antibiotic production. Metabolomics is an emerging technological platform to analyze low molecular weight metabolites in a given organism qualitatively and quantitatively. Compared to other Omics platform, metabolomics has greater advantage in monitoring metabolic flux distribution and thus identifying key metabolites related to target metabolic pathway. The present work aims at establishing a rapid, accurate sample preparation protocol for metabolomics analysis in streptomycetes. In the present work, several sample preparation steps, including cell quenching time, cell separation method, conditions for metabolite extraction and metabolite derivatization were optimized. Then, the metabolic profiles of Streptomyces coelicolor during different growth stages were analyzed by GC-MS. The optimal sample preparation conditions were as follows: time of low-temperature quenching 4 min, cell separation by fast filtration, time of freeze-thaw 45 s/3 min and the conditions of metabolite derivatization at 40 degrees C for 90 min. By using this optimized protocol, 103 metabolites were finally identified from a sample of S. coelicolor, which distribute in central metabolic pathways (glycolysis, pentose phosphate pathway and citrate cycle), amino acid, fatty acid, nucleotide metabolic pathways, etc. By comparing the temporal profiles of these metabolites, the amino acid and fatty acid metabolic pathways were found to stay at a high level during stationary phase, therefore, these pathways may play an important role during the transition between the primary and secondary metabolism. An optimized protocol of sample preparation was established and applied for metabolomics analysis of S. coelicolor, 103 metabolites were identified. The temporal profiles of metabolites reveal amino acid and fatty acid metabolic pathways may play an important role in the transition from primary to

  14. INTERNATIONAL ENVIRONMENTAL TECHNOLOGY IDENTIFICATION, DEVELOPMENT, DEMONSTRATION, DEPLOYMENT AND EXCHANGE

    SciTech Connect

    Roy C. Herndon

    2001-02-28

    Cooperative Agreement (DE-FC21-95EW55101) between the U.S. Department of Energy (DOE) and the Florida State University's Institute for International Cooperative Environmental Research (IICER) was designed to facilitate a number of joint programmatic goals of both the DOE and the IICER related to international technology identification, development, demonstration and deployment using a variety of mechanisms to accomplish these goals. These mechanisms included: laboratory and field research; technology demonstrations; international training and technical exchanges; data collection, synthesis and evaluation; the conduct of conferences, symposia and high-level meetings; and other appropriate and effective approaches. The DOE utilized the expertise and facilities of the IICER at Florida State University to accomplish its goals related to this cooperative agreement. The IICER has unique and demonstrated capabilities that have been utilized to conduct the tasks for this cooperative agreement. The IICER conducted activities related to technology identification, development, evaluation, demonstration and deployment through its joint centers which link the capabilities at Florida State University with collaborating academic and leading research institutions in the major countries of Central and Eastern Europe (e.g., Czech Republic, Hungary, Poland) and Russia. The activities and accomplishments for this five-year cooperative agreement are summarized in this Final Technical Report.

  15. ArgR of Streptomyces coelicolor Is a Versatile Regulator

    PubMed Central

    Pérez-Redondo, Rosario; Rodríguez-García, Antonio; Botas, Alma; Santamarta, Irene; Martín, Juan F.; Liras, Paloma

    2012-01-01

    ArgR is the regulator of arginine biosynthesis genes in Streptomyces species. Transcriptomic comparison by microarrays has been made between Streptomyces coelicolor M145 and its mutant S. coelicolor ΔargR under control, unsupplemented conditions, and in the presence of arginine. Expression of 459 genes was different in transcriptomic assays, but only 27 genes were affected by arginine supplementation. Arginine and pyrimidine biosynthesis genes were derepressed by the lack of ArgR, while no strong effect on expression resulted on arginine supplementation. Several nitrogen metabolism genes expression as glnK, glnA and glnII, were downregulated in S. coelicolor ΔargR. In addition, downregulation of genes for the yellow type I polyketide CPK antibiotic and for the antibiotic regulatory genes afsS and scbR was observed. The transcriptomic data were validated by either reverse transcription-PCR, expression of the gene-promoter coupled to the luciferase gene, proteomic or by electrophoresis mobility shift assay (EMSA) using pure Strep-tagged ArgR. Two ARG-boxes in the arginine operon genes suggest that these genes are more tightly controlled. Other genes, including genes encoding regulatory proteins, possess a DNA sequence formed by a single ARG-box which responds to ArgR, as validated by EMSA. PMID:22403700

  16. Texas LPG fuel cell development and demonstration project

    SciTech Connect

    None, None

    2004-07-26

    The State Energy Conservation Office has executed its first Fuel Cell Project which was awarded under a Department of Energy competitive grant process. The Texas LPG Fuel Processor Development and Fuel Cell Demonstration Program is a broad-based public/private partnership led by the Texas State Energy Conservation Office (SECO). Partners include the Alternative Fuels Research and Education Division (AFRED) of the Railroad Commission of Texas; Plug Power, Inc., Latham, NY, UOP/HyRadix, Des Plaines, IL; Southwest Research Institute (SwRI), San Antonio, TX; the Texas Natural Resource Conservation Commission (TNRCC), and the Texas Department of Transportation (TxDOT). The team proposes to mount a development and demonstration program to field-test and evaluate markets for HyRadix's LPG fuel processor system integrated into Plug Power's residential-scale GenSys(TM) 5C (5 kW) PEM fuel cell system in a variety of building types and conditions of service. The program's primary goal is to develop, test, and install a prototype propane-fueled residential fuel cell power system supplied by Plug Power and HyRadix in Texas. The propane industry is currently funding development of an optimized propane fuel processor by project partner UOP/HyRadix through its national checkoff program, the Propane Education and Research Council (PERC). Following integration and independent verification of performance by Southwest Research Institute, Plug Power and HyRadix will produce a production-ready prototype unit for use in a field demonstration. The demonstration unit produced during this task will be delivered and installed at the Texas Department of Transportation's TransGuide headquarters in San Antonio, Texas. Simultaneously, the team will undertake a market study aimed at identifying and quantifying early-entry customers, technical and regulatory requirements, and other challenges and opportunities that need to be addressed in planning commercialization of the units. For further

  17. Clean coal technologies: Research, development, and demonstration program plan

    SciTech Connect

    Not Available

    1993-12-01

    The US Department of Energy, Office of Fossil Energy, has structured an integrated program for research, development, and demonstration of clean coal technologies that will enable the nation to use its plentiful domestic coal resources while meeting environmental quality requirements. The program provides the basis for making coal a low-cost, environmentally sound energy choice for electric power generation and fuels production. These programs are briefly described.

  18. Demonstration of Decision Support Tools for Sustainable Development

    SciTech Connect

    Shropshire, David Earl; Jacobson, Jacob Jordan; Berrett, Sharon; Cobb, D. A.; Worhach, P.

    2000-11-01

    The Demonstration of Decision Support Tools for Sustainable Development project integrated the Bechtel/Nexant Industrial Materials Exchange Planner and the Idaho National Engineering and Environmental Laboratory System Dynamic models, demonstrating their capabilities on alternative fuel applications in the Greater Yellowstone-Teton Park system. The combined model, called the Dynamic Industrial Material Exchange, was used on selected test cases in the Greater Yellow Teton Parks region to evaluate economic, environmental, and social implications of alternative fuel applications, and identifying primary and secondary industries. The test cases included looking at compressed natural gas applications in Teton National Park and Jackson, Wyoming, and studying ethanol use in Yellowstone National Park and gateway cities in Montana. With further development, the system could be used to assist decision-makers (local government, planners, vehicle purchasers, and fuel suppliers) in selecting alternative fuels, vehicles, and developing AF infrastructures. The system could become a regional AF market assessment tool that could help decision-makers understand the behavior of the AF market and conditions in which the market would grow. Based on this high level market assessment, investors and decision-makers would become more knowledgeable of the AF market opportunity before developing detailed plans and preparing financial analysis.

  19. Development and demonstration of autonomous behaviors for urban environment exploration

    NASA Astrophysics Data System (ADS)

    Ahuja, Gaurav; Fellars, Donald; Kogut, Gregory; Pacis Rius, Estrellina; Schoolov, Misha; Xydes, Alexander

    2012-06-01

    Under the Urban Environment Exploration project, the Space and Naval Warfare Systems Center Pacic (SSC- PAC) is maturing technologies and sensor payloads that enable man-portable robots to operate autonomously within the challenging conditions of urban environments. Previously, SSC-PAC has demonstrated robotic capabilities to navigate and localize without GPS and map the ground oors of various building sizes.1 SSC-PAC has since extended those capabilities to localize and map multiple multi-story buildings within a specied area. To facilitate these capabilities, SSC-PAC developed technologies that enable the robot to detect stairs/stairwells, maintain localization across multiple environments (e.g. in a 3D world, on stairs, with/without GPS), visualize data in 3D, plan paths between any two points within the specied area, and avoid 3D obstacles. These technologies have been developed as independent behaviors under the Autonomous Capabilities Suite, a behavior architecture, and demonstrated at a MOUT site at Camp Pendleton. This paper describes the perceptions and behaviors used to produce these capabilities, as well as an example demonstration scenario.

  20. The development and demonstration of hybrid programmable attitude control electronics

    NASA Technical Reports Server (NTRS)

    Smith, L. S.; Kopf, E. H., Jr.

    1973-01-01

    In the course of extended life attitude control system (ELACS) research sponsored by NASA a hybrid programable attitude control electronics (HYPACE) concept was developed and demonstrated. The wide variety of future planetary missions demanded a new control approach to accommodate the automatic fault tolerance and long the life requirements of such missions. HYPACE provides an adaptable, analog/digital design approach that permits preflight and in-flight accommodation of mission changes, component performance variations, and spacecraft changes, through programing. This enabled broad multimission flexibility of application in a cost effective manner. Previously, flight control computers have not been not flown on planetary missions because of weight and power problems. These problems were resolved in the design of HYPACE. The HYPACE design, which was demonstrated in breadboard form on a single-axis gas-bearing spacecraft simulation, uses a single control channel to perform the attitude control functions sequentially, thus significantly reducing the number of component parts over hard-wired designs.

  1. Phase 1 Development Testing of the Advanced Manufacturing Demonstrator Engine

    NASA Technical Reports Server (NTRS)

    Case, Nicholas L.; Eddleman, David E.; Calvert, Marty R.; Bullard, David B.; Martin, Michael A.; Wall, Thomas R.

    2016-01-01

    The Additive Manufacturing Development Breadboard Engine (BBE) is a pressure-fed liquid oxygen/pump-fed liquid hydrogen (LOX/LH2) expander cycle engine that was built and operated by NASA at Marshall Space Flight Center's East Test Area. The breadboard engine was conceived as a technology demonstrator for the additive manufacturing technologies for an advanced upper stage prototype engine. The components tested on the breadboard engine included an ablative chamber, injector, main fuel valve, turbine bypass valve, a main oxidizer valve, a mixer and the fuel turbopump. All parts minus the ablative chamber were additively manufactured. The BBE was successfully hot fire tested seven times. Data collected from the test series will be used for follow on demonstration tests with a liquid oxygen turbopump and a regeneratively cooled chamber and nozzle.

  2. Environmental signals triggering methylenomycin production by Streptomyces coelicolor A3(2).

    PubMed Central

    Hayes, A; Hobbs, G; Smith, C P; Oliver, S G; Butler, P R

    1997-01-01

    Methylenomycin production by Streptomyces coelicolor A3(2) may be triggered by either of two environmental signals: alanine growth-rate-limiting conditions and/or an acidic pH shock. The production of this SCP1-encoded antibiotic was studied by using batch and chemostat cultures. Batch cultures indicated a role for both nutritional status and culture pH in its regulation. Steady-state methylenomycin production and transcription of an mmy gene under alanine but not glucose growth-rate-limiting conditions was demonstrated in chemostat culture. Transient mmy expression and methylenomycin production occurred following an acidic pH shock. This stimulation of methylenomycin production occurred independently of the nutritional status of the growth environment. Antibiotic production was partially suppressed under alanine compared with glucose growth-rate-limiting conditions following the acidic pH shock. A low specific growth rate was a prerequisite for both steady-state and transient production of methylenomycin. PMID:9287007

  3. Development and demonstration of a telerobotic excavation system

    NASA Technical Reports Server (NTRS)

    Burks, Barry L.; Thompson, David H.; Killough, Stephen M.; Dinkins, Marion A.

    1994-01-01

    Oak Ridge National Laboratory is developing remote excavation technologies for the Department of Energy's Office (DOE) of Technology Development, Robotics Technology Development Program, and also for the Department of Defense (DOD) Project Manager for Ammunition Logistics. This work is being done to meet the need for remote excavation and removal of radioactive and contaminated buried waste at several DOE sites and unexploded ordnance at DOD sites. System requirements are based on the need to uncover and remove waste from burial sites in a way that does not cause unnecessary personnel exposure or additional environmental contamination. Goals for the current project are to demonstrate dexterous control of a backhoe with force feedback and to implement robotic operations that will improve productivity. The Telerobotic Small Emplacement Excavator is a prototype system that incorporates the needed robotic and telerobotic capabilities on a commercially available platform. The ability to add remote dexterous teleoperation and robotic operating modes is intended to be adaptable to other commercially available excavator systems.

  4. The Development of a Demonstration Passive System Reliability Assessment

    SciTech Connect

    Bucknor, Matthew; Grabaskas, David; Brunett, Acacia

    2014-06-22

    In this paper, the details of the development of a demonstration problem to assess the reliability of a passive safety system are presented. An advanced small modular reactor (advSMR) design, which is a pool-type sodium fast reactor (SFR) coupled with a passive reactor cavity cooling system (RCCS) is described. The RELAP5-3D models of the advSMR and RCCS that will be used to simulate a long-term station blackout (SBO) accident scenario are presented. Proposed benchmarking techniques for both the reactor and the RCCS are discussed, which includes utilization of experimental results from the Natural convection Shutdown heat removal Test Facility (NSTF) at the Argonne National Laboratory. Details of how mechanistic methods, specifically the Reliability Method for Passive Systems (RMPS) approach, will be utilized to determine passive system reliability are presented. The results of this mechanistic analysis will ultimately be compared to results from dynamic methods in future work. This work is part of an ongoing project at Argonne to demonstrate methodologies for assessing passive system reliability.

  5. Antibiotic overproduction in Streptomyces coelicolor A3 2 mediated by phosphofructokinase deletion.

    PubMed

    Borodina, Irina; Siebring, Jeroen; Zhang, Jie; Smith, Colin P; van Keulen, Geertje; Dijkhuizen, Lubbert; Nielsen, Jens

    2008-09-12

    Streptomycetes are exploited for production of a wide range of secondary metabolites, and there is much interest in enhancing the level of production of these metabolites. Secondary metabolites are synthesized in dedicated biosynthetic routes, but precursors and co-factors are derived from the primary metabolism. High level production of antibiotics in streptomycetes therefore requires engineering of the primary metabolism. Here we demonstrate this by targeting a key enzyme in glycolysis, phosphofructokinase, leading to improved antibiotic production in Streptomyces coelicolor A3(2). Deletion of pfkA2 (SCO5426), one of three annotated pfkA homologues in S. coelicolor A3(2), resulted in a higher production of the pigmented antibiotics actinorhodin and undecylprodigiosin. The pfkA2 deletion strain had an increased carbon flux through the pentose phosphate pathway, as measured by (13)C metabolic flux analysis, establishing the ATP-dependent PfkA2 as a key player in determining the carbon flux distribution. The increased pentose phosphate pathway flux appeared largely because of accumulation of glucose 6-phosphate and fructose 6-phosphate, as experimentally observed in the mutant strain. Through genome-scale metabolic model simulations, we predicted that decreased phosphofructokinase activity leads to an increase in pentose phosphate pathway flux and in flux to pigmented antibiotics and pyruvate. Integrated analysis of gene expression data using a genome-scale metabolic model further revealed transcriptional changes in genes encoding redox co-factor-dependent enzymes as well as those encoding pentose phosphate pathway enzymes and enzymes involved in storage carbohydrate biosynthesis.

  6. Structure and Function of the RedJ Protein, a Thioesterase from the Prodiginine Biosynthetic Pathway in Streptomyces coelicolor

    SciTech Connect

    Whicher, Jonathan R.; Florova, Galina; Sydor, Paulina K.; Singh, Renu; Alhamadsheh, Mamoun; Challis, Gregory L.; Reynolds, Kevin A.; Smith, Janet L.

    2011-08-17

    Prodiginines are a class of red-pigmented natural products with immunosuppressant, anticancer, and antimalarial activities. Recent studies on prodiginine biosynthesis in Streptomyces coelicolor have elucidated the function of many enzymes within the pathway. However, the function of RedJ, which was predicted to be an editing thioesterase based on sequence similarity, is unknown. We report here the genetic, biochemical, and structural characterization of the redJ gene product. Deletion of redJ in S. coelicolor leads to a 75% decrease in prodiginine production, demonstrating its importance for prodiginine biosynthesis. RedJ exhibits thioesterase activity with selectivity for substrates having long acyl chains and lacking a {beta}-carboxyl substituent. The thioesterase has 1000-fold greater catalytic efficiency with substrates linked to an acyl carrier protein (ACP) than with the corresponding CoA thioester substrates. Also, RedJ strongly discriminates against the streptomycete ACP of fatty acid biosynthesis in preference to RedQ, an ACP of the prodiginine pathway. The 2.12 {angstrom} resolution crystal structure of RedJ provides insights into the molecular basis for the observed substrate selectivity. A hydrophobic pocket in the active site chamber is positioned to bind long acyl chains, as suggested by a long-chain ligand from the crystallization solution bound in this pocket. The accessibility of the active site is controlled by the position of a highly flexible entrance flap. These data combined with previous studies of prodiginine biosynthesis in S. coelicolor support a novel role for RedJ in facilitating transfer of a dodecanoyl chain from one acyl carrier protein to another en route to the key biosynthetic intermediate 2-undecylpyrrole.

  7. Structure and Function of the RedJ Protein, a Thioesterase from the Prodiginine Biosynthetic Pathway in Streptomyces coelicolor*

    PubMed Central

    Whicher, Jonathan R.; Florova, Galina; Sydor, Paulina K.; Singh, Renu; Alhamadsheh, Mamoun; Challis, Gregory L.; Reynolds, Kevin A.; Smith, Janet L.

    2011-01-01

    Prodiginines are a class of red-pigmented natural products with immunosuppressant, anticancer, and antimalarial activities. Recent studies on prodiginine biosynthesis in Streptomyces coelicolor have elucidated the function of many enzymes within the pathway. However, the function of RedJ, which was predicted to be an editing thioesterase based on sequence similarity, is unknown. We report here the genetic, biochemical, and structural characterization of the redJ gene product. Deletion of redJ in S. coelicolor leads to a 75% decrease in prodiginine production, demonstrating its importance for prodiginine biosynthesis. RedJ exhibits thioesterase activity with selectivity for substrates having long acyl chains and lacking a β-carboxyl substituent. The thioesterase has 1000-fold greater catalytic efficiency with substrates linked to an acyl carrier protein (ACP) than with the corresponding CoA thioester substrates. Also, RedJ strongly discriminates against the streptomycete ACP of fatty acid biosynthesis in preference to RedQ, an ACP of the prodiginine pathway. The 2.12 Å resolution crystal structure of RedJ provides insights into the molecular basis for the observed substrate selectivity. A hydrophobic pocket in the active site chamber is positioned to bind long acyl chains, as suggested by a long-chain ligand from the crystallization solution bound in this pocket. The accessibility of the active site is controlled by the position of a highly flexible entrance flap. These data combined with previous studies of prodiginine biosynthesis in S. coelicolor support a novel role for RedJ in facilitating transfer of a dodecanoyl chain from one acyl carrier protein to another en route to the key biosynthetic intermediate 2-undecylpyrrole. PMID:21543318

  8. Development Status of the Fission Power System Technology Demonstration Unit

    NASA Technical Reports Server (NTRS)

    Briggs, Maxwell H.; Gibson, Marc A.; Geng, Steven M.; Pearson, Jon Boise; Godfoy, Thomas

    2012-01-01

    This paper summarizes the progress that has been made in the development of the Fission Power System Technology Demonstration Unit (TDU). The reactor simulator core and Annular Linear Induction Pump have been fabricated and assembled into a test loop at the NASA Marshall Space Flight Center. A 12 kWe Power Conversion Unit (PCU) is being developed consisting of two 6 kWe free-piston Stirling engines. The two 6 kWe engines have been fabricated by Sunpower Inc. and are currently being tested separately prior to integration into the PCU. The Facility Cooling System (FCS) used to reject convertor waste heat has been assembled and tested at the NASA Glenn Research Center (GRC). The structural elements, including a Buildup Assembly Platform (BAP) and Upper Truss Structure (UTS) have been fabricated, and will be used to test cold-end components in thermal vacuum prior to TDU testing. Once all components have been fully tested at the subsystem level, they will be assembled into an end-to-end system and tested in thermal vacuum at GRC.

  9. Development Status of the Fission Power System Technology Demonstration Unit

    NASA Technical Reports Server (NTRS)

    Briggs, Maxwell H.; Gibson, Marc A.; Geng, Steven M; Pearson, Jon Boise; Godfroy, Thomas

    2012-01-01

    This paper summarizes the progress that has been made in the development of the Fission Power System Technology Demonstration Unit (TDU). The reactor simulator core and Annular Linear Induction Pump have been fabricated and assembled into a test loop at the NASA Marshall Space Flight Center. A 12 kWe Power Conversion Unit (PCU) is being developed consisting of two 6 kWe free-piston Stirling engines. The two 6 kWe engines have been fabricated by Sunpower Inc. and are currently being tested separately prior to integration into the PCU. The Facility Cooling System (FCS) used to reject convertor waste heat has been assembled and tested at the NASA Glenn Research Center (GRC). The structural elements, including a Buildup Assembly Platform (BAP) and Upper Truss Structure (UTS) have been fabricated, and will be used to test cold-end components in thermal vacuum prior to TDU testing. Once all components have been fully tested at the subsystem level, they will be assembled into an end-to-end system and tested in thermal vacuum at NASA GRC.

  10. Genetic and Proteomic Analyses of Pupylation in Streptomyces coelicolor

    PubMed Central

    Compton, Corey L.; Fernandopulle, Michael S.; Nagari, Rohith T.

    2015-01-01

    ABSTRACT Pupylation is a posttranslational modification peculiar to actinobacteria wherein proteins are covalently modified with a small protein called the prokaryotic ubiquitin-like protein (Pup). Like ubiquitination in eukaryotes, this phenomenon has been associated with proteasome-mediated protein degradation in mycobacteria. Here, we report studies of pupylation in a streptomycete that is phylogentically related to mycobacteria. We constructed mutants of Streptomyces coelicolor lacking PafA (Pup ligase), the proteasome, and the Pup-proteasome system. We found that these mutants share a high susceptibility to oxidative stress compared to that of the wild-type strain. Remarkably, we found that the pafA null mutant has a sporulation defect not seen in strains lacking the Pup-proteasome system. In proteomics experiments facilitated by an affinity-tagged variant of Pup, we identified 110 pupylated proteins in S. coelicolor strains having and lacking genes encoding the 20S proteasome. Our findings shed new light on this unusual posttranslational modification and its role in Streptomyces physiology. IMPORTANCE The presence of 20S proteasomes reminiscent of those in eukaryotes and a functional equivalent of ubiquitin, known as the prokaryotic ubiquitin-like protein (Pup), in actinobacteria have motivated reevaluations of protein homeostasis in prokaryotes. Though the Pup-proteasome system has been studied extensively in mycobacteria, it is much less understood in streptomycetes, members of a large genus of actinobacteria known for highly choreographed life cycles in which phases of morphological differentiation, sporulation, and secondary metabolism are often regulated by protein metabolism. Here, we define constituents of the pupylome in Streptomyces coelicolor for the first time and present new evidence that links pupylation and the oxidative stress response in this bacterium. Surprisingly, we found that the Pup ligase has a Pup-independent role in sporulation. PMID

  11. Secure, Mobile, Wireless Network Technology Designed, Developed, and Demonstrated

    NASA Technical Reports Server (NTRS)

    Ivancic, William D.; Paulsen, Phillip E.

    2004-01-01

    The inability to seamlessly disseminate data securely over a high-integrity, wireless broadband network has been identified as a primary technical barrier to providing an order-of-magnitude increase in aviation capacity and safety. Secure, autonomous communications to and from aircraft will enable advanced, automated, data-intensive air traffic management concepts, increase National Air Space (NAS) capacity, and potentially reduce the overall cost of air travel operations. For the first time ever, secure, mobile, network technology was designed, developed, and demonstrated with state-ofthe- art protocols and applications by a diverse, cooperative Government-industry team led by the NASA Glenn Research Center. This revolutionary technology solution will make fundamentally new airplane system capabilities possible by enabling secure, seamless network connections from platforms in motion (e.g., cars, ships, aircraft, and satellites) to existing terrestrial systems without the need for manual reconfiguration. Called Mobile Router, the new technology autonomously connects and configures networks as they traverse from one operating theater to another. The Mobile Router demonstration aboard the Neah Bay, a U.S. Coast Guard vessel stationed in Cleveland, Ohio, accomplished secure, seamless interoperability of mobile network systems across multiple domains without manual system reconfiguration. The Neah Bay was chosen because of its low cost and communications mission similarity to low-Earth-orbiting satellite platforms. This technology was successfully advanced from technology readiness level (TRL) 2 (concept and/or application formation) to TRL 6 (system model or prototype demonstration in a relevant environment). The secure, seamless interoperability offered by the Mobile Router and encryption device will enable several new, vehicle-specific and systemwide technologies to perform such things as remote, autonomous aircraft performance monitoring and early detection and

  12. Replisome Localization in Vegetative and Aerial Hyphae of Streptomyces coelicolor

    PubMed Central

    Ruban-Ośmiałowska, Beata; Jakimowicz, Dagmara; Smulczyk-Krawczyszyn, Aleksandra; Chater, Keith F.; Zakrzewska-Czerwińska, Jolanta

    2006-01-01

    Using a functional fusion of DnaN to enhanced green fluorescent protein, we examined the subcellular localization of the replisome machinery in the vegetative mycelium and aerial mycelium of the multinucleoid organism Streptomyces coelicolor. Chromosome replication took place in many compartments of both types of hypha, with the apical compartments of the aerial mycelium exhibiting the highest replication activity. Within a single compartment, the number of “current” ongoing DNA replications was lower than the expected chromosome number, and the appearance of fluorescent foci was often heterogeneous, indicating that this process is asynchronous within compartments and that only selected chromosomes undergo replication. PMID:17015671

  13. OsdR of Streptomyces coelicolor and the Dormancy Regulator DevR of Mycobacterium tuberculosis Control Overlapping Regulons

    PubMed Central

    Urem, Mia; van Rossum, Teunke; Bucca, Giselda; Moolenaar, Geri F.; Laing, Emma; Świątek-Połatyńska, Magda A.; Willemse, Joost; Tenconi, Elodie; Rigali, Sébastien; Goosen, Nora; Smith, Colin P.

    2016-01-01

    ABSTRACT Two-component regulatory systems allow bacteria to respond adequately to changes in their environment. In response to a given stimulus, a sensory kinase activates its cognate response regulator via reversible phosphorylation. The response regulator DevR activates a state of dormancy under hypoxia in Mycobacterium tuberculosis, allowing this pathogen to escape the host defense system. Here, we show that OsdR (SCO0204) of the soil bacterium Streptomyces coelicolor is a functional orthologue of DevR. OsdR, when activated by the sensory kinase OsdK (SCO0203), binds upstream of the DevR-controlled dormancy genes devR, hspX, and Rv3134c of M. tuberculosis. In silico analysis of the S. coelicolor genome combined with in vitro DNA binding studies identified many binding sites in the genomic region around osdR itself and upstream of stress-related genes. This binding correlated well with transcriptomic responses, with deregulation of developmental genes and genes related to stress and hypoxia in the osdR mutant. A peak in osdR transcription in the wild-type strain at the onset of aerial growth correlated with major changes in global gene expression. Taken together, our data reveal the existence of a dormancy-related regulon in streptomycetes which plays an important role in the transcriptional control of stress- and development-related genes. IMPORTANCE Dormancy is a state of growth cessation that allows bacteria to escape the host defense system and antibiotic challenge. Understanding the mechanisms that control dormancy is of key importance for the treatment of latent infections, such as those from Mycobacterium tuberculosis. In mycobacteria, dormancy is controlled by the response regulator DevR, which responds to conditions of hypoxia. Here, we show that OsdR of Streptomyces coelicolor recognizes the same regulatory element and controls a regulon that consists of genes involved in the control of stress and development. Only the core regulon in the direct

  14. Development and Demonstration of Active Noise Control Concepts

    NASA Technical Reports Server (NTRS)

    Kraft, R.; Hu, Z.; Sommerfeldt, S.; Walker, B.; Hersh, A.; Luo, H.; Spencer, M.; Hallman, D.; Mitchell, C.; Sutliff, D.

    2000-01-01

    This report details design methods for and feasibility of an Active Noise Control (ANC) system using flush-wall-mounted sensors and actuators to reduce turbofan engine rotor-stator interaction noise. ANC concepts capable of suppressing discrete-tone spinning modes containing several cut-on radial mode were identified, developed analytically, and evaluated. Separate ANC systems that suppressed at least three radial modes in a cylindrical inlet duct and three radial modes in an exhaust annulus were developed. These designs resulted in inlet duct and exhaust duct tests that were performed at NASA on the 4-ft ANC Fan in the NASA Glenn AAPL facility. Effective suppression of 2-BPF spinning mode m = 2 tone noise was achieved over a range of fan speeds 1800 to 2450 rpm, where up to 4 radials were present. In the inlet duct, up to 12 dB reduction was obtained for 3 radial modes, and up to 4 dB was obtained with 4 radial modes. In the exhaust duct, up to 15 dB PWL reduction was obtained with either two or three radial modes present. Thus, the ability to suppress multiple radial modes for tones in both the inlet and exhaust ducts has been successfully demonstrated. Implications of ANC system design requirements on installation and system integration issues for ANC systems capable of suppressing higher order radial mode content when applied to a 767 using twin CF6 engines were evaluated analytically. The analytical results indicated an ANC system must be part of an integrated design to be effective.

  15. Early Development of Demonstratives in Pre-Qin Chinese

    ERIC Educational Resources Information Center

    Deng, Lin

    2011-01-01

    This dissertation offers a new dynamic account of the evolution of the demonstrative system in pre-Qin Chinese based on a comprehensive linguistic analysis of the phonological, morphological, syntactic, semantic, and pragmatic aspects of demonstratives attested in two corpora of excavated texts, i.e. the oracle-bone inscriptions dated to the late…

  16. New loci required for Streptomyces coelicolor morphological and physiological differentiation.

    PubMed Central

    Champness, W C

    1988-01-01

    Streptomyces coelicolor colonies differentiate both morphologically, producing aerial spore chains, and physiologically, producing antibiotics as secondary metabolites. Single mutations, which block both aspects of differentiation, define bld (bald colony) genes. To identify new bld genes, mutagenized colonies were screened for blocks in the earliest stage of sporulation, the formation of aerial mycelia, and blocks in antibiotic synthesis. The mutations in 12 mutants were mapped; in each strain, the pleiotropic phenotype was due to a single mutation. Seven of the strains contained mutations in known bld loci, bldA and bldB. Three strains contained mutations in a new locus, bldG, and two contained mutations in another new locus, bldH. Like the previously defined bldA mutants, the bldG and bldH mutants were developmentally blocked on glucose. On a variety of carbon sources whose utilization was subject to glucose repression, the developmental blocks were partially relieved for bldG (and bldA) mutants and fully relieved for bldH mutants. These results are compatible with an hypothesis which suggests that there are two alternative controls on S. coelicolor differentiation, one of which is glucose repressible. PMID:3343216

  17. The dynamic architecture of the metabolic switch in Streptomyces coelicolor

    PubMed Central

    2010-01-01

    Background During the lifetime of a fermenter culture, the soil bacterium S. coelicolor undergoes a major metabolic switch from exponential growth to antibiotic production. We have studied gene expression patterns during this switch, using a specifically designed Affymetrix genechip and a high-resolution time-series of fermenter-grown samples. Results Surprisingly, we find that the metabolic switch actually consists of multiple finely orchestrated switching events. Strongly coherent clusters of genes show drastic changes in gene expression already many hours before the classically defined transition phase where the switch from primary to secondary metabolism was expected. The main switch in gene expression takes only 2 hours, and changes in antibiotic biosynthesis genes are delayed relative to the metabolic rearrangements. Furthermore, global variation in morphogenesis genes indicates an involvement of cell differentiation pathways in the decision phase leading up to the commitment to antibiotic biosynthesis. Conclusions Our study provides the first detailed insights into the complex sequence of early regulatory events during and preceding the major metabolic switch in S. coelicolor, which will form the starting point for future attempts at engineering antibiotic production in a biotechnological setting. PMID:20053288

  18. Development and Demonstration of Autonomous Behaviors for Urban Environment Exploration

    DTIC Science & Technology

    2012-04-01

    portable robots to operate autonomously within the challenging conditions of urban environments. Previously, SSC-PAC has demonstrated robotic capa... robot to detect stairs/stairwells, maintain localization across multiple environments (e.g. in a 3D world, on stairs, with/without GPS), visualize data...perceptions and behaviors used to produce these capabilities, as well as an example demonstration scenario. Keywords: robotics , urban environment

  19. The Phosphotransferase System of Streptomyces coelicolor Is Biased for N-Acetylglucosamine Metabolism

    PubMed Central

    Nothaft, Harald; Dresel, Dagmar; Willimek, Andreas; Mahr, Kerstin; Niederweis, Michael; Titgemeyer, Fritz

    2003-01-01

    Mutation of the crr-ptsI gene locus revealed that Streptomyces coelicolor uses the phosphotransferase system (PTS) for N-acetylglucosamine uptake. crr, ptsI, and ptsH, which encode the three general PTS phosphotransferases, are induced by N-acetylglucosamine but not by other PTS substrates. Thus, the S. coelicolor PTS is biased for N-acetylglucosamine utilization, a novel feature that distinguishes this PTS from others. PMID:14617669

  20. Development of image analysis demonstration packages with MENU: educational aspects.

    PubMed

    Hasman, A; Tilanus, C C; Staats, R A

    1984-01-01

    In this paper a tool, MENU, is presented, with which demonstration packages can be easily constructed. The teacher designs the set-up of the package by editing a demonstration specification file, containing both commands to MENU to display frames to the end-user or to execute tasks and the text of the frames. The text contains explanations for the end-user together with the options he can choose. MENU takes care that the corresponding actions are executed. Two image analysis packages, one about CT and one about gated cardiac bloodpool scintigraphy, are presented as examples of the use of MENU. It is concluded, that with MENU (existing) programs can be modeled into packages very easily and efficiently. MENU proves to be a tool that is worthwhile for educational purposes.

  1. New Sporulation Loci in Streptomyces coelicolor A3(2)

    PubMed Central

    Ryding, N. Jamie; Bibb, Maureen J.; Molle, Virginie; Findlay, Kim C.; Chater, Keith F.; Buttner, Mark J.

    1999-01-01

    Sporulation mutants of Streptomyces coelicolor appear white because they are defective in the synthesis of the grey polyketide spore pigment, and such white (whi) mutants had been used to define eight sporulation loci, whiA, whiB, whiD, whiE, whiG, whiH, whiI, and whiJ (K. F. Chater, J. Gen. Microbiol. 72:9–28, 1972; N. J. Ryding, Ph.D. thesis, University of East Anglia, 1995). In an attempt to identify new whi loci, we mutagenized S. coelicolor M145 spores with nitrosoguanidine and identified 770 mutants with colonies ranging from white to medium grey. After excluding unstable strains, we examined the isolates by phase-contrast microscopy and chose 115 whi mutants with clear morphological phenotypes for further study. To exclude mutants representing cloned whi genes, self-transmissible SCP2*-derived plasmids carrying whiA, whiB, whiG, whiH, or whiJ (but not whiD, whiE, or whiI) were introduced into each mutant by conjugation, and strains in which the wild-type phenotype was restored either partially or completely by any of these plasmids were excluded from further analysis. In an attempt to complement some of the remaining 31 whi mutants, an SCP2* library of wild-type S. coelicolor chromosomal DNA was introduced into 19 of the mutants by conjugation. Clones restoring the wild-type phenotype to 12 of the 19 strains were isolated and found to represent five distinct loci, designated whiK, whiL, whiM, whiN, and whiO. Each of the five loci was located on the ordered cosmid library: whiL, whiM, whiN, and whiO occupied positions distinct from previously cloned whi genes; whiK was located on the same cosmid overlap as whiD, but the two loci were shown by complementation to be distinct. The phenotypes resulting from mutations at each of these new loci are described. PMID:10464216

  2. Redox flow cell development and demonstration project, calendar year 1977

    NASA Technical Reports Server (NTRS)

    1979-01-01

    Research and development on the redox flow cell conducted from January 1, 1977, to December 31, 1977, are described in this report. The major focus of the effort during 1977 was the key technology issues that directly influence the fundamental feasibility of the overall redox concept. These issues were the development of a suitable ion exchange membrane for the system, the screening and study of candidate redox couples to achieve optimum cell performance, and the carrying out of systems analysis and modeling to develop system performance goals and cost estimates.

  3. DEMONSTRATION BULLETIN: FLAME REACTOR - HORSEHEAD RESOURCE DEVELOPMENT COMPANY, INC.

    EPA Science Inventory

    The Horsehead Resource Development Company, Inc. (HRD) Flame Reactor is a patented and proven high temperature thermal process designed to safely treat industrial residues and wastes containing metals. During processing, the waste material is introduced into the hottest portio...

  4. Redox flow cell development and demonstration project, calendar year 1976

    NASA Technical Reports Server (NTRS)

    1977-01-01

    The major focus of the effort was the key technology issues that directly influence the fundamental feasibility of the overall redox concept. These issues were the development of a suitable semipermeable separator membrane for the system, the screening and study of candidate redox couples to achieve optimum cell performance, and the carrying out of systems analysis and modeling to develop system performance goals and cost estimates.

  5. Flexible structure control laboratory development and technology demonstration

    NASA Technical Reports Server (NTRS)

    Vivian, H. C.; Blaire, P. E.; Eldred, D. B.; Fleischer, G. E.; Ih, C.-H. C.; Nerheim, N. M.; Scheid, R. E.; Wen, J. T.

    1987-01-01

    An experimental structure is described which was constructed to demonstrate and validate recent emerging technologies in the active control and identification of large flexible space structures. The configuration consists of a large, 20 foot diameter antenna-like flexible structure in the horizontal plane with a gimballed central hub, a flexible feed-boom assembly hanging from the hub, and 12 flexible ribs radiating outward. Fourteen electrodynamic force actuators mounted to the hub and to the individual ribs provide the means to excite the structure and exert control forces. Thirty permanently mounted sensors, including optical encoders and analog induction devices provide measurements of structural response at widely distributed points. An experimental remote optical sensor provides sixteen additional sensing channels. A computer samples the sensors, computes the control updates and sends commands to the actuators in real time, while simultaneously displaying selected outputs on a graphics terminal and saving them in memory. Several control experiments were conducted thus far and are documented. These include implementation of distributed parameter system control, model reference adaptive control, and static shape control. These experiments have demonstrated the successful implementation of state-of-the-art control approaches using actual hardware.

  6. Development and demonstration of flueric sounding rocket motor ignition

    NASA Technical Reports Server (NTRS)

    Marchese, V. P.

    1974-01-01

    An analytical and experimental program is described which established a flueric rocket motor ignition system concept incorporating a pneumatic match with a simple hand pump as the only energy source. An evaluation was made of this concept to determine the margins of the operating range and capabilities of every component of the system. This evaluation included a determination of power supply requirements, ignitor geometry and alinement, ignitor/propellant interfacing and materials and the effects of ambient temperatures and pressure. It was demonstrated that an operator using a simple hand pump for 30 seconds could ignite BKNO3 at a standoff distance of 100 m (330 ft) with the only connection to the ignitor being a piece of plastic pneumatic tubing.

  7. NEW APPROACHES: Serendipity in the development of demonstration experiments

    NASA Astrophysics Data System (ADS)

    Raghuram, P. T.; Kesava Rao, K.

    1996-09-01

    Some experiments drawn from granular mechanics, fluid mechanics and heat and mass transfer are described. These experiments demonstrate the following aspects: (i) the deformation of a dense assembly of particles such as sand leads to an increase in the porosity, (ii) the stress field in a static bed of granular material is non-hydrostatic, (iii) the motion of an hour-glass in a sealed cylinder filled with water is affected by wall friction, (iv) when a syringe containing salt water is lowered into a cylinder filled with fresh water, oscillatory flow is observed due to a convective instability, (v) the `drinking duck' (a toy) works better when water is replaced by a more volatile liquid such as methanol. It is also shown that the accidental use of variations from the published procedures leads to interesting results.

  8. 40 CFR 258.4 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...) SOLID WASTES CRITERIA FOR MUNICIPAL SOLID WASTE LANDFILLS General § 258.4 Research, development, and... include such terms and conditions at least as protective as the criteria for municipal solid waste... and quantities of municipal solid waste and non-hazardous wastes which the State Director...

  9. Research, Development, Demonstration and Deployment Issues in the Power Sector

    EPA Science Inventory

    For Frank Princiotta’s book, Global Climate Change—The Technology Challenge In this chapter we explore the challenges in developing and deploying technology for mitigation of CO2 emissions associated with power generation. Past successes with controlling other pollutants (notab...

  10. 40 CFR 258.4 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ...) SOLID WASTES CRITERIA FOR MUNICIPAL SOLID WASTE LANDFILLS General § 258.4 Research, development, and... include such terms and conditions at least as protective as the criteria for municipal solid waste... and quantities of municipal solid waste and non-hazardous wastes which the State Director...

  11. Development of a Thermoacoustic Stirling Engine Technology Demonstrator

    NASA Astrophysics Data System (ADS)

    Reissner, Alexander; Gerger, Joachim; Hummel, Stefan; Reißig, Jannis; Pawelke, Roland

    2014-08-01

    Waste heat is a primary source of energy loss in many aerospace and terrestrial applications. FOTEC, an Austrian Research Company located in Wiener Neustadt, is presently developing a micro power converter, promising high efficiencies even for small- scale applications. The converter is based on an innovative thermoacoustic stirling engine concept without any moving parts. Such a maintenance-free engine system would be particularly suitable for advanced space power systems (radioisotope, waste heat) or even within the scope of terrestrial energy harvesting. This paper will summarizes the status of our ongoing efforts on this micro power converter technology.

  12. Automotive Fuel Processor Development and Demonstration with Fuel Cell Systems

    SciTech Connect

    Nuvera Fuel Cells

    2005-04-15

    The potential for fuel cell systems to improve energy efficiency and reduce emissions over conventional power systems has generated significant interest in fuel cell technologies. While fuel cells are being investigated for use in many applications such as stationary power generation and small portable devices, transportation applications present some unique challenges for fuel cell technology. Due to their lower operating temperature and non-brittle materials, most transportation work is focusing on fuel cells using proton exchange membrane (PEM) technology. Since PEM fuel cells are fueled by hydrogen, major obstacles to their widespread use are the lack of an available hydrogen fueling infrastructure and hydrogen's relatively low energy storage density, which leads to a much lower driving range than conventional vehicles. One potential solution to the hydrogen infrastructure and storage density issues is to convert a conventional fuel such as gasoline into hydrogen onboard the vehicle using a fuel processor. Figure 2 shows that gasoline stores roughly 7 times more energy per volume than pressurized hydrogen gas at 700 bar and 4 times more than liquid hydrogen. If integrated properly, the fuel processor/fuel cell system would also be more efficient than traditional engines and would give a fuel economy benefit while hydrogen storage and distribution issues are being investigated. Widespread implementation of fuel processor/fuel cell systems requires improvements in several aspects of the technology, including size, startup time, transient response time, and cost. In addition, the ability to operate on a number of hydrocarbon fuels that are available through the existing infrastructure is a key enabler for commercializing these systems. In this program, Nuvera Fuel Cells collaborated with the Department of Energy (DOE) to develop efficient, low-emission, multi-fuel processors for transportation applications. Nuvera's focus was on (1) developing fuel processor

  13. Development and Flight Demonstration of Space Debris Monitor (SDM)

    NASA Astrophysics Data System (ADS)

    Kitazawa, Yukihito; Hanada, Toshiya; Matsumoto, Haruhisa; Kobayashi, Masanori; Sakurai, Akira; Yasaka, Tetsuo; Funakoshi, Kunihiro; Hasegawa, Sunao; Akahoshi, Yasuhiro; Kimoto, Yugo; Okudaira, Osamu; Kamiya, Koki; Nakamura, Maki

    2016-07-01

    The space debris monitor (SDM) is a large-area impact sensor for in situ measurements of micro-meteoroids and space debris of the sub-millimeter to millimeter size in the near-Earth space environment. These meteoroid and debris particles are very small to be detected by ground-based observations (radars and optical telescopes) but are sufficiently large to cause serious damage to spacecraft equipment in the low Earth orbit region. The nominal detection area of the SDM is 0.1 m^2 (0.35 m × 0.3 m), but its dimensions can be easily modified to accommodate different SDM constraints. The SDM is made from a flexible printed circuit, which is produced from a thin film of a nonconductive material (such as polyimide) on which thin conductive stripes are formed in parallel. The stripe width is approximately 50 μm, and the spatial separation is approximately 100 μm, as shown in Figure 1. When a micro-debris particle with an effective diameter near to or larger than the spatial separation of the stripes (here approximately 100 μm) collides with the sensor film at a velocity sufficient to penetrate it, one or more of the stripes are cut and become nonconductive. Debris impacts can thus be detected by monitoring the electrical conductivity (resistivity) of the stripes. This sensor system can measure the size of the incident micro-debris particles by detecting the number of severed stripes. The measurement concept is registered as a patent in many countries. The first SDM was launched with HTV-5 on August 19, 2015 and represented the world's first micro-debris measurement demonstration experiment to be conducted on the ISS using the concept of conductive (resistive) strip lines for real-time debris detection.

  14. Development and Demonstration of an Ada Test Generation System

    NASA Technical Reports Server (NTRS)

    1996-01-01

    important role in identifying testing requirements which are infeasible. This is especially true for data flow testing and modified condition/decision coverage. Our system uses in an essential way symbolic analysis and theorem proving technology, and we believe this work represents one of the few successful uses of a theorem prover working in a completely automatic fashion to solve a problem of practical interest. We believe this work anticipates an important trend away from purely syntactic-based methods for program analysis to semantic methods based on symbolic processing and inference technology. Other results demonstrating the practical use of automatic inference is being reported in hardware verification, although there are significant differences between the hardware work and ours. However, what is common and important is that general purpose theorem provers are being integrated with more special-purpose decision procedures to solve problems in analysis and verification. We are pursuina commercial opportunities for this work, and will use and extend the work in other projects we are engaged in. Ultimately we would like to rework the system to analyze C, C++, or Java as a key step toward commercialization.

  15. The development and testing of pulsed detonation engine ground demonstrators

    NASA Astrophysics Data System (ADS)

    Panicker, Philip Koshy

    2008-10-01

    The successful implementation of a PDE running on fuel and air mixtures will require fast-acting fuel-air injection and mixing techniques, detonation initiation techniques such as DDT enhancing devices or a pre-detonator, an effective ignition system that can sustain repeated firing at high rates and a fast and capable, closed-loop control system. The control system requires high-speed transducers for real-time monitoring of the PDE and the detection of the detonation wave speed. It is widely accepted that the detonation properties predicted by C-J detonation relations are fairly accurate in comparison to experimental values. The post-detonation flow properties can also be expressed as a function of wave speed or Mach number. Therefore, the PDE control system can use C-J relations to predict the post-detonation flow properties based on measured initial conditions and compare the values with those obtained from using the wave speed. The controller can then vary the initial conditions within the combustor for the subsequent cycle, by modulating the frequency and duty cycle of the valves, to obtain optimum air and fuel flow rates, as well as modulate the energy and timing of the ignition to achieve the required detonation properties. Five different PDE ground demonstrators were designed, built and tested to study a number of the required sub-systems. This work presents a review of all the systems that were tested, along with suggestions for their improvement. The PDE setups, ranged from a compact PDE with a 19 mm (3/4 in.) i.d., to two 25 mm (1 in.) i.d. setups, to a 101 mm (4 in.) i.d. dual-stage PDE setup with a pre-detonator. Propane-oxygen mixtures were used in the smaller PDEs. In the dual-stage PDE, propane-oxygen was used in the pre-detonator, while propane-air mixtures were used in the main combustor. Both rotary valves and solenoid valve injectors were studied. The rotary valves setups were tested at 10 Hz, while the solenoid valves were tested at up to 30 Hz

  16. The Coordinated Positive Regulation of Topoisomerase Genes Maintains Topological Homeostasis in Streptomyces coelicolor

    PubMed Central

    Gongerowska, Martyna; Gutkowski, Paweł; Zakrzewska-Czerwińska, Jolanta; Jakimowicz, Dagmara

    2016-01-01

    ABSTRACT Maintaining an optimal level of chromosomal supercoiling is critical for the progression of DNA replication and transcription. Moreover, changes in global supercoiling affect the expression of a large number of genes and play a fundamental role in adapting to stress. Topoisomerase I (TopA) and gyrase are key players in the regulation of bacterial chromosomal topology through their respective abilities to relax and compact DNA. Soil bacteria such as Streptomyces species, which grow as branched, multigenomic hyphae, are subject to environmental stresses that are associated with changes in chromosomal topology. The topological fluctuations modulate the transcriptional activity of a large number of genes and in Streptomyces are related to the production of antibiotics. To better understand the regulation of topological homeostasis in Streptomyces coelicolor, we investigated the interplay between the activities of the topoisomerase-encoding genes topA and gyrBA. We show that the expression of both genes is supercoiling sensitive. Remarkably, increased chromosomal supercoiling induces the topA promoter but only slightly influences gyrBA transcription, while DNA relaxation affects the topA promoter only marginally but strongly activates the gyrBA operon. Moreover, we showed that exposure to elevated temperatures induces rapid relaxation, which results in changes in the levels of both topoisomerases. We therefore propose a unique mechanism of S. coelicolor chromosomal topology maintenance based on the supercoiling-dependent stimulation, rather than repression, of the transcription of both topoisomerase genes. These findings provide important insight into the maintenance of topological homeostasis in an industrially important antibiotic producer. IMPORTANCE We describe the unique regulation of genes encoding two topoisomerases, topoisomerase I (TopA) and gyrase, in a model Streptomyces species. Our studies demonstrate the coordination of topoisomerase gene

  17. Large-Scale Transposition Mutagenesis of Streptomyces coelicolor Identifies Hundreds of Genes Influencing Antibiotic Biosynthesis.

    PubMed

    Xu, Zhong; Wang, Yemin; Chater, Keith F; Ou, Hong-Yu; Xu, H Howard; Deng, Zixin; Tao, Meifeng

    2017-03-15

    Gram-positive Streptomyces bacteria produce thousands of bioactive secondary metabolites, including antibiotics. To systematically investigate genes affecting secondary metabolism, we developed a hyperactive transposase-based Tn5 transposition system and employed it to mutagenize the model species Streptomyces coelicolor, leading to the identification of 51,443 transposition insertions. These insertions were distributed randomly along the chromosome except for some preferred regions associated with relatively low GC content in the chromosomal core. The base composition of the insertion site and its flanking sequences compiled from the 51,443 insertions implied a 19-bp expanded target site surrounding the insertion site, with a slight nucleic acid base preference in some positions, suggesting a relative randomness of Tn5 transposition targeting in the high-GC Streptomyces genome. From the mutagenesis library, 724 mutants involving 365 genes had altered levels of production of the tripyrrole antibiotic undecylprodigiosin (RED), including 17 genes in the RED biosynthetic gene cluster. Genetic complementation revealed that most of the insertions (more than two-thirds) were responsible for the changed antibiotic production. Genes associated with branched-chain amino acid biosynthesis, DNA metabolism, and protein modification affected RED production, and genes involved in signaling, stress, and transcriptional regulation were overrepresented. Some insertions caused dramatic changes in RED production, identifying future targets for strain improvement.IMPORTANCE High-GC Gram-positive streptomycetes and related actinomycetes have provided more than 100 clinical drugs used as antibiotics, immunosuppressants, and antitumor drugs. Their genomes harbor biosynthetic genes for many more unknown compounds with potential as future drugs. Here we developed a useful genome-wide mutagenesis tool based on the transposon Tn5 for the study of secondary metabolism and its regulation

  18. Impact of Malic Enzymes on Antibiotic and Triacylglycerol Production in Streptomyces coelicolor

    PubMed Central

    Navone, Laura; Casati, Paula; Gramajo, Hugo

    2012-01-01

    In this paper, we have characterized two malic enzymes (ME), SCO2951 and SCO5261, from Streptomyces coelicolor and analyzed their role in antibiotic and triacylglycerol (TAG) production. Biochemical studies have demonstrated that Sco2951 and Sco5261 genes encode NAD+- and NADP+-dependent malic enzymes, respectively. Single or double mutants in the ME-encoding genes show no effect on growth rate compared to the parental M145 strain. However, the single Sco2951 and the double Sco2951 Sco5261 mutants display a strong reduction in the production of the polyketide antibiotic actinorhodin; additionally, the Sco2951 Sco5261 mutant shows a decrease in stored TAGs during exponential growth. The lower production of actinorhodin in the double mutant occurs as a consequence of a decrease in the expression of actII-ORF4, the transcriptional activator of the actinorhodin gene cluster. On the other hand, the reduced TAG accumulation is not due to reduced transcript levels of fatty acid biosynthetic genes nor to changes in the amount of the precursor acetyl coenzyme A (acetyl-CoA). This mutant accumulates intermediates of the tricarboxylic acid (TCA) cycle that could alter the regulation of the actinorhodin biosynthetic pathway, suggesting that MEs are important anaplerotic enzymes that redirect C4 intermediates from the TCA cycle to maintain secondary metabolism and TAG production in Streptomyces. PMID:22544242

  19. Evidence for the negative regulation of phytase gene expression in Streptomyces lividans and Streptomyces coelicolor.

    PubMed

    Boukhris, Ines; Dulermo, Thierry; Chouayekh, Hichem; Virolle, Marie-Joëlle

    2016-01-01

    Sco7697, a gene encoding a phytase, enzyme able to degrade phytate (myo-inositol 1,2,3,4,5,6-hexakis phosphate), the most abundant phosphorus storing compound in plants is present in the genome of S. coelicolor, a soil born bacteria with a saprophytic lifestyle. The expression of this gene was previously shown to be induced in conditions of Pi limitation by the response regulator PhoP binding to an operator sequence, the PHO box, located upstream of the -35 promoter sequence. A close examination of the promoter region of sco7697 revealed the presence of another putative operator site, a Direct Repeat (DR), located downstream of the -10 promoter sequence. In order to determine whether this DR played a role in regulation of sco7697 expression, different variants of the phytase gene promoter region were transcriptionally fused to the ß-glucuronidase reporter gene (GUS). As expected, deletion of the PHO box led to abolition of sco7697 induction in conditions of Pi limitation. Interestingly, alteration of the DR correlated with a dramatic increase of GUS expression but only when PhoP was present. These results demonstrated that this DR is the site of strong negative regulation by an unknown repressor. The latter would impede the necessary activation of phytase expression by PhoP.

  20. Phosphoprotein affinity purification identifies proteins involved in S-adenosyl-L-methionine-induced enhancement of antibiotic production in Streptomyces coelicolor.

    PubMed

    Meng, Lingzhu; Yang, Seung Hwan; Palaniyandi, Sasikumar Arunachalam; Lee, Sung-Kwon; Lee, In-Ae; Kim, Tae-Jong; Suh, Joo-Won

    2011-01-01

    Streptomycetes are the major natural source of clinical antibiotics. The enhanced secondary metabolite production of many streptomycetes by S-adenosylmethionine (SAM) in previous studies suggested the existence of a common SAM regulatory effect. We screened nine proteins using the phosphoprotein purification column from Streptomyces coelicolor. Among them, genes (SCO5477, SCO5113, SCO4647, SCO4885 and SCO1793) for five proteins were disrupted by insertion mutation. The undecylprodigiosin and actinorhodin productions were changed in all mutations. The SAM-induced enhancement of actinorhodin production was abolished by all mutations except SCO4885 mutation, which reduced the production of actinorhodin and undecylprodigiosin with SAM treatment. This study demonstrates that phosphoprotein affinity purification can be used as a screening method to identify the proteins involved SAM signaling.

  1. The dynamic transcriptional and translational landscape of the model antibiotic producer Streptomyces coelicolor A3(2)

    PubMed Central

    Jeong, Yujin; Kim, Ji-Nu; Kim, Min Woo; Bucca, Giselda; Cho, Suhyung; Yoon, Yeo Joon; Kim, Byung-Gee; Roe, Jung-Hye; Kim, Sun Chang; Smith, Colin P.; Cho, Byung-Kwan

    2016-01-01

    Individual Streptomyces species have the genetic potential to produce a diverse array of natural products of commercial, medical and veterinary interest. However, these products are often not detectable under laboratory culture conditions. To harness their full biosynthetic potential, it is important to develop a detailed understanding of the regulatory networks that orchestrate their metabolism. Here we integrate nucleotide resolution genome-scale measurements of the transcriptome and translatome of Streptomyces coelicolor, the model antibiotic-producing actinomycete. Our systematic study determines 3,570 transcription start sites and identifies 230 small RNAs and a considerable proportion (∼21%) of leaderless mRNAs; this enables deduction of genome-wide promoter architecture. Ribosome profiling reveals that the translation efficiency of secondary metabolic genes is negatively correlated with transcription and that several key antibiotic regulatory genes are translationally induced at transition growth phase. These findings might facilitate the design of new approaches to antibiotic discovery and development. PMID:27251447

  2. Comparative genomic hybridizations reveal absence of large Streptomyces coelicolor genomic islands in Streptomyces lividans

    PubMed Central

    Jayapal, Karthik P; Lian, Wei; Glod, Frank; Sherman, David H; Hu, Wei-Shou

    2007-01-01

    Background The genomes of Streptomyces coelicolor and Streptomyces lividans bear a considerable degree of synteny. While S. coelicolor is the model streptomycete for studying antibiotic synthesis and differentiation, S. lividans is almost exclusively considered as the preferred host, among actinomycetes, for cloning and expression of exogenous DNA. We used whole genome microarrays as a comparative genomics tool for identifying the subtle differences between these two chromosomes. Results We identified five large S. coelicolor genomic islands (larger than 25 kb) and 18 smaller islets absent in S. lividans chromosome. Many of these regions show anomalous GC bias and codon usage patterns. Six of them are in close vicinity of tRNA genes while nine are flanked with near perfect repeat sequences indicating that these are probable recent evolutionary acquisitions into S. coelicolor. Embedded within these segments are at least four DNA methylases and two probable methyl-sensing restriction endonucleases. Comparison with S. coelicolor transcriptome and proteome data revealed that some of the missing genes are active during the course of growth and differentiation in S. coelicolor. In particular, a pair of methylmalonyl CoA mutase (mcm) genes involved in polyketide precursor biosynthesis, an acyl-CoA dehydrogenase implicated in timing of actinorhodin synthesis and bldB, a developmentally significant regulator whose mutation causes complete abrogation of antibiotic synthesis belong to this category. Conclusion Our findings provide tangible hints for elucidating the genetic basis of important phenotypic differences between these two streptomycetes. Importantly, absence of certain genes in S. lividans identified here could potentially explain the relative ease of DNA transformations and the conditional lack of actinorhodin synthesis in S. lividans. PMID:17623098

  3. Role of Acid Metabolism in Streptomyces coelicolor Morphological Differentiation and Antibiotic Biosynthesis

    PubMed Central

    Viollier, Patrick H.; Minas, Wolfgang; Dale, Glenn E.; Folcher, Marc; Thompson, Charles J.

    2001-01-01

    Studies of citrate synthase (CitA) were carried out to investigate its role in morphological development and biosynthesis of antibiotics in Streptomyces coelicolor. Purification of CitA, the major vegetative enzyme activity, allowed characterization of its kinetic properties. The apparent Km values of CitA for acetyl coenzyme A (acetyl-CoA) (32 μM) and oxaloacetate (17 μM) were similar to those of citrate synthases from other gram-positive bacteria and eukaryotes. CitA was not strongly inhibited by various allosteric feedback inhibitors (NAD+, NADH, ATP, ADP, isocitrate, or α-ketoglutarate). The corresponding gene (citA) was cloned and sequenced, allowing construction of a citA mutant (BZ2). BZ2 was a glutamate auxotroph, indicating that citA encoded the major citrate synthase allowing flow of acetyl-CoA into the tricarboxylic acid (TCA) cycle. Interruption of aerobic TCA cycle-based metabolism resulted in acidification of the medium and defects in morphological differentiation and antibiotic biosynthesis. These developmental defects of the citA mutant were in part due to a glucose-dependent medium acidification that was also exhibited by some other bald mutants. Unlike other acidogenic bald strains, citA and bldJ mutants were able to produce aerial mycelia and pigments when the medium was buffered sufficiently to maintain neutrality. Extracellular complementation studies suggested that citA defines a new stage of the Streptomyces developmental cascade. PMID:11325948

  4. Role of acid metabolism in Streptomyces coelicolor morphological differentiation and antibiotic biosynthesis.

    PubMed

    Viollier, P H; Minas, W; Dale, G E; Folcher, M; Thompson, C J

    2001-05-01

    Studies of citrate synthase (CitA) were carried out to investigate its role in morphological development and biosynthesis of antibiotics in Streptomyces coelicolor. Purification of CitA, the major vegetative enzyme activity, allowed characterization of its kinetic properties. The apparent K(m) values of CitA for acetyl coenzyme A (acetyl-CoA) (32 microM) and oxaloacetate (17 microM) were similar to those of citrate synthases from other gram-positive bacteria and eukaryotes. CitA was not strongly inhibited by various allosteric feedback inhibitors (NAD(+), NADH, ATP, ADP, isocitrate, or alpha-ketoglutarate). The corresponding gene (citA) was cloned and sequenced, allowing construction of a citA mutant (BZ2). BZ2 was a glutamate auxotroph, indicating that citA encoded the major citrate synthase allowing flow of acetyl-CoA into the tricarboxylic acid (TCA) cycle. Interruption of aerobic TCA cycle-based metabolism resulted in acidification of the medium and defects in morphological differentiation and antibiotic biosynthesis. These developmental defects of the citA mutant were in part due to a glucose-dependent medium acidification that was also exhibited by some other bald mutants. Unlike other acidogenic bald strains, citA and bldJ mutants were able to produce aerial mycelia and pigments when the medium was buffered sufficiently to maintain neutrality. Extracellular complementation studies suggested that citA defines a new stage of the Streptomyces developmental cascade.

  5. A technical platform for generating reproducible expression data from Streptomyces coelicolor batch cultivations.

    PubMed

    Battke, F; Herbig, A; Wentzel, A; Jakobsen, O M; Bonin, M; Hodgson, D A; Wohlleben, W; Ellingsen, T E; Nieselt, K

    2011-01-01

    Streptomyces coelicolor, the model species of the genus Streptomyces, presents a complex life cycle of successive morphological and biochemical changes involving the formation of substrate and aerial mycelium, sporulation and the production of antibiotics. The switch from primary to secondary metabolism can be triggered by nutrient starvation and is of particular interest as some of the secondary metabolites produced by related Streptomycetes are commercially relevant. To understand these events on a molecular basis, a reliable technical platform encompassing reproducible fermentation as well as generation of coherent transcriptomic data is required. Here, we investigate the technical basis of a previous study as reported by Nieselt et al. (BMC Genomics 11:10, 2010) in more detail, based on the same samples and focusing on the validation of the custom-designed microarray as well as on the reproducibility of the data generated from biological replicates. We show that the protocols developed result in highly coherent transcriptomic measurements. Furthermore, we use the data to predict chromosomal gene clusters, extending previously known clusters as well as predicting interesting new clusters with consistent functional annotations.

  6. Regulation of a novel gene cluster involved in secondary metabolite production in Streptomyces coelicolor.

    PubMed

    Hindra; Pak, Patricia; Elliot, Marie A

    2010-10-01

    Antibiotic biosynthesis in the streptomycetes is a complex and highly regulated process. Here, we provide evidence for the contribution of a novel genetic locus to antibiotic production in Streptomyces coelicolor. The overexpression of a gene cluster comprising four protein-encoding genes (abeABCD) and an antisense RNA-encoding gene (α-abeA) stimulated the production of the blue-pigmented metabolite actinorhodin on solid medium. Actinorhodin production also was enhanced by the overexpression of an adjacent gene (abeR) encoding a predicted Streptomyces antibiotic regulatory protein (SARP), while the deletion of this gene impaired actinorhodin production. We found the abe genes to be differentially regulated and controlled at multiple levels. Upstream of abeA was a promoter that directed the transcription of abeABCD at a low but constitutive level. The expression of abeBCD was, however, significantly upregulated at a time that coincided with the initiation of aerial development and the onset of secondary metabolism; this expression was activated by the binding of AbeR to four heptameric repeats upstream of a promoter within abeA. Expressed divergently to the abeBCD promoter was α-abeA, whose expression mirrored that of abeBCD but did not require activation by AbeR. Instead, α-abeA transcript levels were subject to negative control by the double-strand-specific RNase, RNase III.

  7. Angucyclines as signals modulate the behaviors of Streptomyces coelicolor.

    PubMed

    Wang, Weishan; Ji, Junjie; Li, Xiao; Wang, Juan; Li, Shanshan; Pan, Guohui; Fan, Keqiang; Yang, Keqian

    2014-04-15

    The angucycline antibiotic jadomycin B (JdB) produced by Streptomyces venezuelae has been found here to induce complex survival responses in Streptomyces coelicolor at subinhibitory concentration. The receptor for JdB was identified as a "pseudo" gamma-butyrolactone receptor, ScbR2, which was shown to bind two previously unidentified target promoters, those of redD (redDp) and adpA (adpAp), thus directly regulating undecylprodigiosin (Red) production and morphological differentiation, respectively. Because AdpA also directly regulates the expression of redD, ScbR2, AdpA, and RedD together form a feed-forward loop controlling both differentiation and Red production phenotypes. Different signal strengths (i.e., JdB concentrations) were shown to induce the two different phenotypes by modulating the relative transcription levels of adpA vs. redD. The induction of morphological differentiation and endogenous antibiotic production by exogenous antibiotic exemplifies an important survival strategy more sophisticated than the induction of antibiotic resistance.

  8. Microbial Transformation of Antibiotics: Phosphorylation of Clindamycin by Streptomyces coelicolor Müller1

    PubMed Central

    Coats, John H.; Argoudelis, Alexander D.

    1971-01-01

    Addition of clindamycin to whole-cell cultures of Streptomyces coelicolor Müller resulted in the loss of in vitro activity against organisms sensitive to clindamycin. Incubation of such culture filtrates with alkaline phosphatase generated a biologically active material identified as clindamycin. Fermentation broths containing inactivated clindamycin yielded clindamycin 3-phosphate, the structure of which was established by physical-chemical and enzymatic studies. Clindamycin was phosphorylated by lysates and partially purified enzyme preparations from S. coelicolor Müller. These reactions require a ribonucleoside triphosphate and Mg2+. The product of the cell-free reactions was identified as clindamycin 3-phosphate. PMID:5166238

  9. Regulation of the dnaK operon of Streptomyces coelicolor A3(2) is governed by HspR, an autoregulatory repressor protein.

    PubMed Central

    Bucca, G; Hindle, Z; Smith, C P

    1997-01-01

    The dnaK operon of Streptomyces coelicolor contains four genes (5'-dnaK-grpE-dnaJ-hspR). The fourth gene encodes a novel heat shock protein, HspR, which appears so far to be unique to the high-G+C actinomycete group of bacteria. HspR binds with high specificity to three inverted repeat sequences in the promoter region of the S. coelicolor dnaK operon, strongly suggesting a direct role for HspR in heat shock gene regulation. Here we present genetic and biochemical evidence that HspR is the repressor of the dnaK operon. Disruption of hspR leads to high-level constitutive transcription of the dnaK operon. Parallel transcriptional analyses of groESL1 and groEL2 expression demonstrated that heat shock regulation of the groE genes was essentially unaffected in an hspR null mutant, although the basal (uninduced) level of groEL2 transcription was slightly elevated compared with the wild type. The results of HspR titration experiments, where the dnaK operon promoter region was cloned at ca. 50 copies per chromosome, were consistent with the prediction that HspR functions as a negative autoregulator. His-tagged HspR, overproduced and purified from Escherichia coli, was shown to repress transcription from the dnaK operon promoter in vitro, providing additional evidence for the proposal that HspR directly regulates transcription of the dnaK operon. These studies indicate that there are at least two transcriptional mechanisms for controlling heat shock genes in S. coelicolor--one controlling the dnaK operon and another controlling the groE genes. PMID:9324243

  10. Differential proteomic analysis of an engineered Streptomyces coelicolor strain reveals metabolic pathways supporting growth on n-hexadecane.

    PubMed

    Gallo, Giuseppe; Lo Piccolo, Luca; Renzone, Giovanni; La Rosa, Ruggero; Scaloni, Andrea; Quatrini, Paola; Puglia, Anna Maria

    2012-06-01

    The alkB gene, encoding an alkane monooxygenase in the actinomycete Gordonia sp. SoCg, was expressed in the non-alkane-degrading actinomycete Streptomyces coelicolor M145. The resulting engineered strain, M145-AH, can grow on n-hexadecane as sole carbon source. To unravel proteins associated with growth on n-alkanes, proteome of M145-AH after 6, 24, and 48 h of incubation in the Bushnell-Haas (BH) mineral medium containing n-hexadecane as sole carbon source (H condition) and in BH without any carbon source (0 condition) were compared using 2D-differential gel electrophoresis. Proteome analysis revealed significant changes only at 48 h, showing 48 differentially abundant proteins identified by mass spectrometry procedures. To asses if these proteins were specifically related to n-hexadecane metabolism, their expression was investigated, comparing H proteome with that of M145-AH incubated in BH with glucose as sole carbon source (G condition). Thus, protein expression profiles at 6, 24, and 48 h under H, 0, and G conditions were combined, revealing that M145-AH regulates in a temporally- and carbon source-dependent manner the expression of proteins involved in regulatory events, central carbon metabolism, respiration, β-oxidation, membrane transport, and amino acid and protein metabolism. Interestingly, 21 % of them, mostly involved in membrane transport and protein metabolism, showed a n-hexadecane-dependent regulation with regulatory proteins such as CRP likely to have a key role in M145-AH n-hexadecane growth. These results, expanding the knowledge on n-alkane utilization in Gram-positive bacteria, reveal genes to be targeted to develop an efficient S. coelicolor M145-AH-based bioremediation system.

  11. Crystal Structure of the Streptomyces coelicolor Sortase E1 Transpeptidase Provides Insight into the Binding Mode of the Novel Class E Sorting Signal

    PubMed Central

    Kattke, Michele D.; Chan, Albert H.; Duong, Andrew; Sexton, Danielle L.; Sawaya, Michael R.; Cascio, Duilio; Elliot, Marie A.; Clubb, Robert T.

    2016-01-01

    Many species of Gram-positive bacteria use sortase transpeptidases to covalently affix proteins to their cell wall or to assemble pili. Sortase-displayed proteins perform critical and diverse functions for cell survival, including cell adhesion, nutrient acquisition, and morphological development, among others. Based on their amino acid sequences, there are at least six types of sortases (class A to F enzymes); however, class E enzymes have not been extensively studied. Class E sortases are used by soil and freshwater-dwelling Actinobacteria to display proteins that contain a non-canonical LAXTG sorting signal, which differs from 90% of known sorting signals by substitution of alanine for proline. Here we report the first crystal structure of a class E sortase, the 1.93 Å resolution structure of the SrtE1 enzyme from Streptomyces coelicolor. The active site is bound to a tripeptide, providing insight into the mechanism of substrate binding. SrtE1 possesses β3/β4 and β6/β7 active site loops that contact the LAXTG substrate and are structurally distinct from other classes. We propose that SrtE1 and other class E sortases employ a conserved tyrosine residue within their β3/β4 loop to recognize the amide nitrogen of alanine at position P3 of the sorting signal through a hydrogen bond, as seen here. Incapability of hydrogen-bonding with canonical proline-containing sorting signals likely contributes to class E substrate specificity. Furthermore, we demonstrate that surface anchoring of proteins involved in aerial hyphae formation requires an N-terminal segment in SrtE1 that is presumably positioned within the cytoplasm. Combined, our results reveal unique features within class E enzymes that enable them to recognize distinct sorting signals, and could facilitate the development of substrate-based inhibitors of this important enzyme family. PMID:27936128

  12. Development and demonstration of manufacturing processes for fabricating graphite/LARC 160 polyimide structural elements

    NASA Technical Reports Server (NTRS)

    Frost, R. K.; Jones, J. S.; Dynes, P. J.; Wykes, D. H.

    1981-01-01

    The development and demonstration of manufacturing technologies for the structural application of Celion graphite/LARC-160 polyimide composite material is discussed. Process development and fabrication of demonstration components are discussed. Process development included establishing quality assurance of the basic composite material and processing, nondestructive inspection of fabricated components, developing processes for specific structural forms, and qualification of processes through mechanical testing. Demonstration components were fabricated. The demonstration components consisted of flat laminates, skin/stringer panels, honeycomb panels, chopped fiber compression moldings, and a technology demonstrator segment (TDS) representative of the space shuttle aft body flap.

  13. WhiD and WhiB, Homologous Proteins Required for Different Stages of Sporulation in Streptomyces coelicolor A3(2)

    PubMed Central

    Molle, Virginie; Palframan, Wendy J.; Findlay, Kim C.; Buttner, Mark J.

    2000-01-01

    The whiD locus, which is required for the differentiation of Streptomyces coelicolor aerial hyphae into mature spore chains, was localized by map-based cloning to the overlap between cosmids 6G4 and D63 of the minimal ordered library of Redenbach et al. (M. Redenbach et al., Mol. Microbiol. 21:77–96, 1996). Subcloning and sequencing showed that whiD encodes a homologue of WhiB, a protein required for the initiation of sporulation septation in S. coelicolor. WhiD and WhiB belong to a growing family of small (76- to 112-residue) proteins of unknown biochemical function in which four cysteines are absolutely conserved; all known members of this family are found in the actinomycetes. A constructed whiD null mutant showed reduced levels of sporulation, and those spores that did form were heat sensitive, lysed extensively, and were highly irregular in size, arising at least in part from irregularity in septum placement. The whiD null mutant showed extreme variation in spore cell wall deposition; most spores had uniformly thin (20- to 30-nm) walls, but spore chains were frequently observed in which there was irregular but very pronounced (up to 170 nm) cell wall thickening at the junctions between spores. whiD null mutant spores were frequently partitioned into irregular smaller units through the deposition of additional septa, which were often laid down in several different planes, very close to the spore poles. These “minicompartments” appeared to be devoid of chromosomal DNA. Two whiD promoters, whiDp1 and whiDp2, were identified, and their activities were analyzed during development of wild-type S. coelicolor on solid medium. Both promoters were developmentally regulated; whiDp1 and whiDp2 transcripts were detected transiently, approximately at the time when sporulation septa were observed in the aerial hyphae. PMID:10671449

  14. Adenosine deaminase from Streptomyces coelicolor: recombinant expression, purification and characterization.

    PubMed

    Pornbanlualap, Somchai; Chalopagorn, Pornchanok

    2011-08-01

    The sequencing of the genome of Streptomyces coelicolor A3(2) identified seven putative adenine/adenosine deaminases and adenosine deaminase-like proteins, none of which have been biochemically characterized. This report describes recombinant expression, purification and characterization of SCO4901 which had been annotated in data bases as a putative adenosine deaminase. The purified putative adenosine deaminase gives a subunit Mr=48,400 on denaturing gel electrophoresis and an oligomer molecular weight of approximately 182,000 by comparative gel filtration. These values are consistent with the active enzyme being composed of four subunits with identical molecular weights. The turnover rate of adenosine is 11.5 s⁻¹ at 30 °C. Since adenine is deaminated ∼10³ slower by the enzyme when compared to that of adenosine, these data strongly show that the purified enzyme is an adenosine deaminase (ADA) and not an adenine deaminase (ADE). Other adenine nucleosides/nucleotides, including 9-β-D-arabinofuranosyl-adenine (ara-A), 5'-AMP, 5'-ADP and 5'-ATP, are not substrates for the enzyme. Coformycin and 2'-deoxycoformycin are potent competitive inhibitors of the enzyme with inhibition constants of 0.25 and 3.4 nM, respectively. Amino acid sequence alignment of ScADA with ADAs from other organisms reveals that eight of the nine highly conserved catalytic site residues in other ADAs are also conserved in ScADA. The only non-conserved residue is Asn317, which replaces Asp296 in the murine enzyme. Based on these data, it is suggested here that ADA and ADE proteins are divergently related enzymes that have evolved from a common α/β barrel scaffold to catalyze the deamination of different substrates, using a similar catalytic mechanism.

  15. Gene-Enzyme Relations of Tryptophan Mutants in STREPTOMYCES COELICOLOR A3(2)

    PubMed Central

    Smithers, Charles M.; Engel, Paulinus P.

    1974-01-01

    Mutations in twenty-eight tryptophan mutants of S. coelicolor A3(2) were mapped relative to the nearest flanking markers. Mutants lacking single enzymatic activities for phosphoribosyltransferase, phosphoribosylanthranilate isomerase, indodeglycerol phosphate synthase, tryptophan synthase A and tryptophan synthase B were identified. PMID:4452474

  16. Exploitation of the Streptomyces coelicolor A3(2) genome sequence for discovery of new natural products and biosynthetic pathways.

    PubMed

    Challis, Gregory L

    2014-02-01

    Streptomyces, and related genera of Actinobacteria, are renowned for their ability to produce antibiotics and other bioactive natural products with a wide range of applications in medicine and agriculture. Streptomyces coelicolor A3(2) is a model organism that has been used for more than five decades to study the genetic and biochemical basis for the production of bioactive metabolites. In 2002, the complete genome sequence of S. coelicolor was published. This greatly accelerated progress in understanding the biosynthesis of metabolites known or suspected to be produced by S. coelicolor and revealed that streptomycetes have far greater potential to produce bioactive natural products than suggested by classical bioassay-guided isolation studies. In this article, efforts to exploit the S. coelicolor genome sequence for the discovery of novel natural products and biosynthetic pathways are summarized.

  17. Structural and functional basis of transcriptional regulation by TetR family protein CprB from S. coelicolor A3(2)

    PubMed Central

    Bhukya, Hussain; Bhujbalrao, Ruchika; Bitra, Aruna; Anand, Ruchi

    2014-01-01

    Antibiotic production and resistance pathways in Streptomyces are dictated by the interplay of transcriptional regulatory proteins that trigger downstream responses via binding to small diffusible molecules. To decipher the mode of DNA binding and the associated allosteric mechanism in the sub-class of transcription factors that are induced by γ-butyrolactones, we present the crystal structure of CprB in complex with the consensus DNA element to a resolution of 3.25 Å. Binding of the DNA results in the restructuring of the dimeric interface of CprB, inducing a pendulum-like motion of the helix-turn-helix motif that inserts into the major groove. The crystal structure revealed that, CprB is bound to DNA as a dimer of dimers with the mode of binding being analogous to the broad spectrum multidrug transporter protein QacR from the antibiotic resistant strain Staphylococcus aureus. It was demonstrated that the CprB displays a cooperative mode of DNA binding, following a clamp and click model. Experiments performed on a subset of DNA sequences from Streptomyces coelicolor A3(2) suggest that CprB is most likely a pleiotropic regulator. Apart from serving as an autoregulator, it is potentially a part of a network of proteins that modulates the γ-butyrolactone synthesis and antibiotic regulation pathways in S. coelicolor A3(2). PMID:25092919

  18. Structural and functional basis of transcriptional regulation by TetR family protein CprB from S. coelicolor A3(2).

    PubMed

    Bhukya, Hussain; Bhujbalrao, Ruchika; Bitra, Aruna; Anand, Ruchi

    2014-09-01

    Antibiotic production and resistance pathways in Streptomyces are dictated by the interplay of transcriptional regulatory proteins that trigger downstream responses via binding to small diffusible molecules. To decipher the mode of DNA binding and the associated allosteric mechanism in the sub-class of transcription factors that are induced by γ-butyrolactones, we present the crystal structure of CprB in complex with the consensus DNA element to a resolution of 3.25 Å. Binding of the DNA results in the restructuring of the dimeric interface of CprB, inducing a pendulum-like motion of the helix-turn-helix motif that inserts into the major groove. The crystal structure revealed that, CprB is bound to DNA as a dimer of dimers with the mode of binding being analogous to the broad spectrum multidrug transporter protein QacR from the antibiotic resistant strain Staphylococcus aureus. It was demonstrated that the CprB displays a cooperative mode of DNA binding, following a clamp and click model. Experiments performed on a subset of DNA sequences from Streptomyces coelicolor A3(2) suggest that CprB is most likely a pleiotropic regulator. Apart from serving as an autoregulator, it is potentially a part of a network of proteins that modulates the γ-butyrolactone synthesis and antibiotic regulation pathways in S. coelicolor A3(2).

  19. Genome-wide analysis of in vivo binding of the master regulator DasR in Streptomyces coelicolor identifies novel non-canonical targets.

    PubMed

    Świątek-Połatyńska, Magdalena A; Bucca, Giselda; Laing, Emma; Gubbens, Jacob; Titgemeyer, Fritz; Smith, Colin P; Rigali, Sébastien; van Wezel, Gilles P

    2015-01-01

    Streptomycetes produce a wealth of natural products, including over half of all known antibiotics. It was previously demonstrated that N-acetylglucosamine and secondary metabolism are closely entwined in streptomycetes. Here we show that DNA recognition by the N-acetylglucosamine-responsive regulator DasR is growth-phase dependent, and that DasR can bind to sites in the S. coelicolor genome that have no obvious resemblance to previously identified DasR-responsive elements. Thus, the regulon of DasR extends well beyond what was previously predicted and includes a large number of genes with functions far removed from N-acetylglucosamine metabolism, such as genes for small RNAs and DNA transposases. Conversely, the DasR regulon during vegetative growth largely correlates to the presence of canonical DasR-responsive elements. The changes in DasR binding in vivo following N-acetylglucosamine induction were studied in detail and a possible molecular mechanism by which the influence of DasR is extended is discussed. Discussion of DasR binding was further informed by a parallel transcriptome analysis of the respective cultures. Evidence is provided that DasR binds directly to the promoters of all genes encoding pathway-specific regulators of antibiotic production in S. coelicolor, thereby providing an exquisitely simple link between nutritional control and secondary metabolism.

  20. Deletion of the signalling molecule synthase ScbA has pleiotropic effects on secondary metabolite biosynthesis, morphological differentiation and primary metabolism in Streptomyces coelicolor A3(2).

    PubMed

    D'Alia, Davide; Eggle, Daniela; Nieselt, Kay; Hu, Wei-Shou; Breitling, Rainer; Takano, Eriko

    2011-03-01

    Streptomycetes have high biotechnological relevance as producers of diverse metabolites widely used in medical and agricultural applications. The biosynthesis of these metabolites is controlled by signalling molecules, γ-butyrolactones, that act as bacterial hormones. In Streptomyces coelicolor, a group of signalling molecules called SCBs (S. coelicolorbutanolides) regulates production of the pigmented antibiotics coelicolor polyketide (CPK), actinorhodin and undecylprodigiosin. The γ-butyrolactone synthase ScbA is responsible for the biosynthesis of SCBs. Here we show the results of a genome-wide transcriptome analysis of a scbA deletion mutant prior to and during the transition to antibiotic production. We report a strong perturbation in the expression of three pigmented antibiotic clusters in the mutant throughout the growth curve, thus providing a molecular explanation for the antibiotic phenotype observed previously. Our study also revealed, for the first time, that the secondary metabolite cluster responsible for synthesis of the siderophore desferrioxamine is under the control of SCB signalling. Moreover, expression of the genes encoding enzymes for primary metabolism pathways, which supply antibiotic precursors and genes for morphological differentiation, was found shifted earlier in time in the mutant. In conclusion, our time series analysis demonstrates new details of the regulatory effects of the γ-butyrolactone system in Streptomyces.

  1. RNA Polymerase Sigma Factor That Blocks Morphological Differentiation by Streptomyces coelicolor

    PubMed Central

    Gehring, Amy M.; Yoo, Narie J.; Losick, Richard

    2001-01-01

    The filamentous bacterium Streptomyces coelicolor undergoes a complicated process of morphological differentiation that begins with the formation of an aerial mycelium and culminates in sporulation. Genes required for the initiation of aerial mycelium formation have been termed bld (bald), describing the smooth, undifferentiated colonies of mutant strains. By using an insertional mutagenesis protocol that relies on in vitro transposition, we have isolated a bld mutant harboring an insertion in a previously uncharacterized gene, SCE59.12c, renamed here rsuA. The insertion mutant exhibited no measurable growth defect but failed to produce an aerial mycelium and showed a significant delay in the production of the polyketide antibiotic actinorhodin. The rsuA gene encodes an apparent anti-sigma factor and is located immediately downstream of SCE59.13c, renamed here sigU, whose product is inferred to be a member of the extracytoplasmic function subfamily of RNA polymerase sigma factors. The absence of rsuA in a strain that contained sigU caused a block in development, and the overexpression of sigU in an otherwise wild-type strain caused a delay in aerial mycelium formation. However, a strain in which both rsuA and sigU had been deleted was able to undergo morphological differentiation normally. We conclude that the rsuA-encoded anti-sigma factor is responsible for antagonizing the function of the sigma factor encoded by sigU. We also conclude that the sigU-encoded sigma factor is not normally required for development but that its uncontrolled activity obstructs morphological differentiation at an early stage. PMID:11566999

  2. A central regulator of morphological differentiation in the multicellular bacterium Streptomyces coelicolor.

    PubMed

    Nguyen, Kien T; Willey, Joanne M; Nguyen, Liem D; Nguyen, Lieu T; Viollier, Patrick H; Thompson, Charles J

    2002-12-01

    In the multicellular bacterium Streptomyces coelicolor, functions of developmental (bald) genes are required for the biosynthesis of SapB, a hydrophobic peptidic morphogen that facilitates aerial hyphae formation. Here, we show that aerial hyphal growth and SapB biosynthesis could be activated independently from the normal developmental cascade by providing unprogrammed expression of functionally interactive genes within the ram cluster. ramC, ramS and ramR were essential for normal growth of aerial hyphae, and ramR, a response regulator gene, was a key activator of development. The ramR gene restored growth of aerial hyphae and SapB formation in all bald strains tested (albeit only weakly in the bldC mutant), many of which are characterized by physiological defects. Disruption of the ramR gene abolished SapB biosynthesis and severely delayed growth of aerial hyphae. Transcription of ramR was developmentally controlled, and RamR function in vivo depended on its putative phosphorylation site (D53). We identified and mapped RamR targets immediately upstream of the region encoding ramC and ramS, a putative operon. Overexpression of ramR in the wild-type strain increased SapB levels and caused a distinctive wrinkled surface topology. Based on these results, we propose that phenotypes of bald mutations reflect an early stage in the Streptomyces developmental programme similar to the spo0 mutations in the unicellular bacterium Bacillus subtilis, and that RamR has analogies to Spo0A, the Bacillus response regulator that integrates physiological signals before triggering endospore formation.

  3. Resuscitation-promoting factors are cell wall-lytic enzymes with important roles in the germination and growth of Streptomyces coelicolor.

    PubMed

    Sexton, Danielle L; St-Onge, Renée J; Haiser, Henry J; Yousef, Mary R; Brady, Lauren; Gao, Chan; Leonard, Jacqueline; Elliot, Marie A

    2015-03-01

    Dormancy is a common strategy adopted by bacterial cells as a means of surviving adverse environmental conditions. For Streptomyces bacteria, this involves developing chains of dormant exospores that extend away from the colony surface. Both spore formation and subsequent spore germination are tightly controlled processes, and while significant progress has been made in understanding the underlying regulatory and enzymatic bases for these, there are still significant gaps in our understanding. One class of proteins with a potential role in spore-associated processes are the so-called resuscitation-promoting factors, or Rpfs, which in other actinobacteria are needed to restore active growth to dormant cell populations. The model species Streptomyces coelicolor encodes five Rpf proteins (RpfA to RfpE), and here we show that these proteins have overlapping functions during growth. Collectively, the S. coelicolor Rpfs promote spore germination and are critical for growth under nutrient-limiting conditions. Previous studies have revealed structural similarities between the Rpf domain and lysozyme, and our in vitro biochemical assays revealed various levels of peptidoglycan cleavage capabilities for each of these five Streptomyces enzymes. Peptidoglycan remodeling by enzymes such as these must be stringently governed so as to retain the structural integrity of the cell wall. Our results suggest that one of the Rpfs, RpfB, is subject to a unique mode of enzymatic autoregulation, mediated by a domain of previously unknown function (DUF348) located within the N terminus of the protein; removal of this domain led to significantly enhanced peptidoglycan cleavage.

  4. The master regulator PhoP coordinates phosphate and nitrogen metabolism, respiration, cell differentiation and antibiotic biosynthesis: comparison in Streptomyces coelicolor and Streptomyces avermitilis.

    PubMed

    Martín, Juan F; Rodríguez-García, Antonio; Liras, Paloma

    2017-03-15

    Phosphate limitation is important for production of antibiotics and other secondary metabolites in Streptomyces. Phosphate control is mediated by the two-component system PhoR-PhoP. Following phosphate depletion, PhoP stimulates expression of genes involved in scavenging, transport and mobilization of phosphate, and represses the utilization of nitrogen sources. PhoP reduces expression of genes for aerobic respiration and activates nitrate respiration genes. PhoP activates genes for teichuronic acid formation and reduces expression of genes for phosphate-rich teichoic acid biosynthesis. In Streptomyces coelicolor, PhoP repressed several differentiation and pleiotropic regulatory genes, which affects development and indirectly antibiotic biosynthesis. A new bioinformatics analysis of the putative PhoP-binding sequences in Streptomyces avermitilis was made. Many sequences in S. avermitilis genome showed high weight values and were classified according to the available genetic information. These genes encode phosphate scavenging proteins, phosphate transporters and nitrogen metabolism genes. Among of the genes highlighted in the new studies was aveR, located in the avermectin gene cluster, encoding a LAL-type regulator, and afsS, which is regulated by PhoP and AfsR. The sequence logo for S. avermitilis PHO boxes is similar to that of S. coelicolor, with differences in the weight value for specific nucleotides in the sequence.The Journal of Antibiotics advance online publication, 15 March 2017; doi:10.1038/ja.2017.19.

  5. Precision Information Environment (PIE) for International Safeguards: Pre-Demonstration Development Use Cases

    SciTech Connect

    Gastelum, Zoe N.; Henry, Michael J.

    2013-11-13

    In FY2013, the PIE International Safeguards team demonstrated our development progress to U.S. Department of Energy (DOE) staff from the Office of Nonproliferation and International Security (NA-24, our client) and the Office of Defense Nuclear Nonproliferation Research and Development (NA-22). Following the demonstration, the team was asked by our client to complete additional development prior to a planned demonstration at the International Atomic Energy Agency (IAEA), scheduled tentatively for January or spring of 2014. The team discussed four potential areas for development (in priority order), and will develop them as time and funding permit prior to an IAEA demonstration. The four capability areas are: 1. Addition of equipment manuals to PIE-accessible files 2. Optical character recognition (OCR) of photographed text 3. Barcode reader with information look-up from a database 4. Add Facilities to Data Model 5. Geospatial capabilities with information integration Each area will be described below in a use case.

  6. A Plan for the Development and Demonstration of Optical Communications for Deep Space

    NASA Technical Reports Server (NTRS)

    Lesh, J. R.; Deutsch, L. J.; Weber, W. J.

    1990-01-01

    In this article, an overall plan for the development and demonstration of optical communications for deep-space applications is presented. The current state of the technology for optical communications is presented. Then, the development and demonstration plan is presented in two parts: the overall major systems activities, followed by the generic technology developments that will enable them. The plan covers the path from laboratory subsystems demonstrations out to a full-scale flight experiment system for the proposed Mars Communications Relay Orbiter mission.

  7. Seeing Is Believing: Creating a Catalyst for Teacher Change through a Demonstration Classroom Professional Development Initiative

    ERIC Educational Resources Information Center

    Grierson, Arlene L.; Gallagher, Tiffany L.

    2009-01-01

    This qualitative case study explored the experiences of eight observing teachers, a demonstration teacher, and a program department consultant, who over a nine-month period participated in a demonstration classroom professional development initiative. Analyses of participants' experiences documented how the initiative created a catalyst for…

  8. Tn4563 transposition in Streptomyces coelicolor and its application to isolation of new morphological mutants.

    PubMed Central

    Schauer, A T; Nelson, A D; Daniel, J B

    1991-01-01

    The Tn3-like transposon Tn4556 (and its derivatives Tn4560 and Tn4563) has been used for insertion mapping of genetic loci cloned on plasmids, but it has been difficult to obtain chromosomal insertions, largely because of the lack of a strong selection against transposon donor molecules. In this communication, we report two efficient selection techniques for transposition and their use in the isolation of chromosomal insertion mutations. A number of independent Streptomyces coelicolor morphological mutants (bld and whi) were obtained. Two of the bld mutations were mapped to locations on the chromosome by SCP1-mediated conjugation; at least one mutation, bld-5m1, appears to define a novel locus involved in control of S. coelicolor morphogenesis and antibiotic production. Images PMID:1650343

  9. Post-translational Serine/Threonine Phosphorylation and Lysine Acetylation: A Novel Regulatory Aspect of the Global Nitrogen Response Regulator GlnR in S. coelicolor M145

    PubMed Central

    Amin, Rafat; Franz-Wachtel, Mirita; Tiffert, Yvonne; Heberer, Martin; Meky, Mohamed; Ahmed, Yousra; Matthews, Arne; Krysenko, Sergii; Jakobi, Marco; Hinder, Markus; Moore, Jane; Okoniewski, Nicole; Maček, Boris; Wohlleben, Wolfgang; Bera, Agnieszka

    2016-01-01

    Soil-dwelling Streptomyces bacteria such as S.coelicolor have to constantly adapt to the nitrogen (N) availability in their habitat. Thus, strict transcriptional and post-translational control of the N-assimilation is fundamental for survival of this species. GlnR is a global response regulator that controls transcription of the genes related to the N-assimilation in S. coelicolor and other members of the Actinomycetales. GlnR represents an atypical orphan response regulator that is not activated by the phosphorylation of the conserved aspartate residue (Asp 50). We have applied transcriptional analysis, LC-MS/MS analysis and electrophoretic mobility shift assays (EMSAs) to understand the regulation of GlnR in S. coelicolor M145. The expression of glnR and GlnR-target genes was revisited under four different N-defined conditions and a complex N-rich condition. Although, the expression of selected GlnR-target genes was strongly responsive to changing N-concentrations, the glnR expression itself was independent of the N-availability. Using LC-MS/MSanalysis we demonstrated that GlnR was post-translationally modified. The post-translational modifications of GlnR comprise phosphorylation of the serine/threonine residues and acetylation of lysine residues. In the complex N-rich medium GlnR was phosphorylated on six serine/threonine residues and acetylated on one lysine residue. Under defined N-excess conditions only two phosphorylated residues were detected whereas under defined N-limiting conditions no phosphorylation was observed. GlnR phosphorylation is thus clearly correlated with N-rich conditions. Furthermore, GlnR was acetylated on four lysine residues independently of the N-concentration in the defined media and on only one lysine residue in the complex N-rich medium. Using EMSAs we demonstrated that phosphorylation inhibited the binding of GlnR to its targets genes, whereas acetylation had little influence on the formation of GlnR-DNA complex. This study clearly

  10. Purification of an Extracellular Signaling Molecule Involved in Production of Aerial Mycelium by Streptomyces coelicolor

    PubMed Central

    Nodwell, Justin R.; Losick, Richard

    1998-01-01

    We have extensively purified a factor from conditioned medium that restores aerial mycelium formation to a mutant of Streptomyces coelicolor that is defective in morphological differentiation. Response to this factor is shown to depend on the presence of the BldK oligopeptide import system. We suggest that this substance acts at the first step in a putative cascade of developmental regulatory signals. PMID:9495776

  11. Uptake of chitosan-derived D-glucosamine oligosaccharides in Streptomyces coelicolor A3(2).

    PubMed

    Viens, Pascal; Dubeau, Marie-Pierre; Kimura, Akane; Desaki, Yoshitake; Shinya, Tomonori; Shibuya, Naoto; Saito, Akihiro; Brzezinski, Ryszard

    2015-05-01

    The csnR gene, localized at the beginning of an operon, csnR-K, which organization is conserved through many actinomycete genomes, was previously shown to repress the transcription of the chitosanase gene csnA in Streptomyces lividans. However, knowledge on the function of the whole csnR-K operon in the metabolism of chitosan (an N-deacetylated derivative of chitin) remained limited. Mutants of S. coelicolor A3(2) harboring partial or total deletions of the csnR-K operon were analyzed for their capacity to uptake glucosamine oligosaccharides (GlcN)n. The csnR-K operon was autoregulated by CsnR repressor and its transcription was inducible by GlcN oligosaccharides. The operon controlled the uptake of GlcN oligosaccharides in S. coelicolor A3(2), with a minor contribution to the consumption of monomeric GlcN but not chitin-related N-acetylated derivatives. The deletion of the whole operon abolished the uptake of GlcN oligosaccharides. The CsnEFG transporter encoded by this operon is the front door for the assimilation of chitosan-derived hydrolysis products in S. coelicolor A3(2). The ATP-binding component MsiK was essential for CsnEFG transport function. Also, deletion of msiK abolished the induction of csnA transcription by GlcN oligosaccharides.

  12. Genome-wide identification and characterization of reference genes with different transcript abundances for Streptomyces coelicolor

    PubMed Central

    Li, Shanshan; Wang, Weishan; Li, Xiao; Fan, Keqiang; Yang, Keqian

    2015-01-01

    The lack of reliable reference genes (RGs) in the genus Streptomyces hampers effort to obtain the precise data of transcript levels. To address this issue, we aimed to identify reliable RGs in the model organism Streptomyces coelicolor. A pool of potential RGs containing 1,471 genes was first identified by determining the intersection of genes with stable transcript levels from four time-series transcriptome microarray datasets of S. coelicolor M145 cultivated in different conditions. Then, following a strict rational selection scheme including homology analysis, disturbance analysis, function analysis and transcript abundance analysis, 13 candidates were selected from the 1,471 genes. Based on real-time quantitative reverse transcription PCR assays, SCO0710, SCO6185, SCO1544, SCO3183 and SCO4758 were identified as the top five genes with the most stable transcript levels among the 13 candidates. Further analyses showed these five genes also maintained stable transcript levels in different S. coelicolor strains, as well as in Streptomyces avermitilis MA-4680 and Streptomyces clavuligerus NRRL 3585, suggesting they could fulfill the requirements of accurate data normalization in streptomycetes. Moreover, the systematic strategy employed in this work could be used for reference in other microorganism to select reliable RGs. PMID:26527303

  13. Research, Development and Demonstration Center in Education of Handicapped Children. Annual Report (1972-1973).

    ERIC Educational Resources Information Center

    Minnesota Univ., Minneapolis. Research, Development, and Demonstration Center in Education of Handicapped Children.

    The University of Minnesota Research, Development, and Demonstration Center in Education of Handicapped Children focuses on intervention strategies and materials to develop and improve communication skills of handicapped children from birth to 9 years of age. Center objectives include identification from longitudinal data of school children with…

  14. Economic Development Projects and Jobs: Lessons from the Targeted Jobs Demonstration Program.

    ERIC Educational Resources Information Center

    Van Horn, Carl; And Others

    This guide, based on approaches for targeting jobs and business opportunities that were developed during the Targeted Jobs Demonstration Program (TJDP), contains strategies and techniques for ensuring that some of the benefits of economic development investments are directed to low-income individuals and small and minority businesses. Addressed in…

  15. Freihoelser Forst Local Training Area Demonstration Project: Prescription development and installation

    SciTech Connect

    Hinchman, R.R.; Zellmer, S.D.; Severinghaus, W.D.; Brent, J.J.

    1989-04-01

    The Freiholser Forst Local Training Area (LTA) Rehabilitation Demonstration Project is part of the Integrated Training Area Management program being developed by the US Army Corps of Engineers` Construction Engineering Research Laboratory for the Seventh Army Training Command of the US Army in Europe. The rehabilitation demonstration project was begun in 1987 to develop and demonstrate rapid, cost-effective methods to stabilize the LTA`s barren, eroding maneuver areas and make training conditions more realistic. The sandy, infertile, and acidic soils at the LTA are considered the major factor limiting rehabilitation efforts there. The project involves the evaluation of three procedures to revegetate the soils, each incorporating identical methods for preparing the seedbed and a single seed mixture consisting of adapted, native species but using different soil amendments. All three treatments have satisfactorily reestablished vegetation and controlled erosion on the demonstration plots at the LTA, but their costs have varied widely.

  16. Freihoelser Forst Local Training Area Demonstration Project: Prescription development and installation

    SciTech Connect

    Hinchman, R.R.; Zellmer, S.D. . Energy Systems Div.); Severinghaus, W.D. ); Brent, J.J. )

    1989-04-01

    The Freiholser Forst Local Training Area (LTA) Rehabilitation Demonstration Project is part of the Integrated Training Area Management program being developed by the US Army Corps of Engineers' Construction Engineering Research Laboratory for the Seventh Army Training Command of the US Army in Europe. The rehabilitation demonstration project was begun in 1987 to develop and demonstrate rapid, cost-effective methods to stabilize the LTA's barren, eroding maneuver areas and make training conditions more realistic. The sandy, infertile, and acidic soils at the LTA are considered the major factor limiting rehabilitation efforts there. The project involves the evaluation of three procedures to revegetate the soils, each incorporating identical methods for preparing the seedbed and a single seed mixture consisting of adapted, native species but using different soil amendments. All three treatments have satisfactorily reestablished vegetation and controlled erosion on the demonstration plots at the LTA, but their costs have varied widely.

  17. Developmental Control of Stress Stimulons in Streptomyces coelicolor Revealed by Statistical Analyses of Global Gene Expression Patterns

    PubMed Central

    Vohradsky, J.; Li, X.-M.; Dale, G.; Folcher, M.; Nguyen, L.; Viollier, P. H.; Thompson, C. J.

    2000-01-01

    Stress-induced regulatory networks coordinated with a procaryotic developmental program were revealed by two-dimensional gel analyses of global gene expression. Four developmental stages were identified by their distinctive protein synthesis patterns using principal component analysis. Statistical analyses focused on five stress stimulons (induced by heat, cold, salt, ethanol, or antibiotic shock) and their synthesis during development. Unlike other bacteria, for which various stresses induce expression of similar sets of protein spots, in Streptomyces coelicolor heat, salt, and ethanol stimulons were composed of independent sets of proteins. This suggested independent control by different physiological stress signals and their corresponding regulatory systems. These stress proteins were also under developmental control. Cluster analysis of stress protein synthesis profiles identified 10 different developmental patterns or “synexpression groups.” Proteins induced by cold, heat, or salt shock were enriched in three developmental synexpression groups. In addition, certain proteins belonging to the heat and salt shock stimulons were coregulated during development. Thus, stress regulatory systems controlling these stimulons were implicated as integral parts of the developmental program. This correlation suggested that thermal shock and salt shock stress response regulatory systems either allow the cell to adapt to stresses associated with development or directly control the developmental program. PMID:10940043

  18. Development and Demonstration of a 25 Watt Thermophotovoltaic Power Source for a Hybrid Power System

    NASA Technical Reports Server (NTRS)

    Doyle, Edward; Shukla, Kailash; Metcalfe, Christopher

    2001-01-01

    The development of a propane-fueled, 25 W thermophotovoltaic (TPV) power source for use in a hybrid power system is described. The TPV power source uses a platinum emitting surface with an anti-reflective coating to radiate to gallium antimonide photocells, which converts the radiation to electric power. The development program started with the design and fabrication of an engineering prototype system. This was used as a component development vehicle to develop the technologies for the various components. A 25 W demonstration prototype was then designed and fabricated using the most advanced component approaches. The designs and test results from this development program are discussed.

  19. A novel insertion mutation in Streptomyces coelicolor ribosomal S12 protein results in paromomycin resistance and antibiotic overproduction.

    PubMed

    Wang, Guojun; Inaoka, Takashi; Okamoto, Susumu; Ochi, Kozo

    2009-03-01

    We identified a novel paromomycin resistance-associated mutation in rpsL, caused by the insertion of a glycine residue at position 92, in Streptomyces coelicolor ribosomal protein S12. This insertion mutation (GI92) resulted in a 20-fold increase in the paromomycin resistance level. In combination with another S12 mutation, K88E, the GI92 mutation markedly enhanced the production of the blue-colored polyketide antibiotic actinorhodin and the red-colored antibiotic undecylprodigiosin. The gene replacement experiments demonstrated that the K88E-GI92 double mutation in the rpsL gene was responsible for the marked enhancement of antibiotic production observed. Ribosomes with the K88E-GI92 double mutation were characterized by error restrictiveness (i.e., hyperaccuracy). Using a cell-free translation system, we found that mutant ribosomes harboring the K88E-GI92 double mutation but not ribosomes harboring the GI92 mutation alone displayed sixfold greater translation activity relative to that of the wild-type ribosomes at late growth phase. This resulted in the overproduction of actinorhodin, caused by the transcriptional activation of the pathway-specific regulatory gene actII-orf4, possibly due to the increased translation of transcripts encoding activators of actII-orf4. The mutant with the K88E-GI92 double mutation accumulated a high level of ribosome recycling factor at late stationary phase, underlying the high level of protein synthesis activity observed.

  20. Induction of antimicrobial activities in heterologous streptomycetes using alleles of the Streptomyces coelicolor gene absA1.

    PubMed

    McKenzie, Nancy L; Thaker, Maulik; Koteva, Kalinka; Hughes, Donald W; Wright, Gerard D; Nodwell, Justin R

    2010-04-01

    The bacterial genus Streptomyces is endowed with a remarkable secondary metabolism that generates an enormous number of bioactive small molecules. Many of these genetically encoded small molecules are used as antibiotics, anticancer agents and as other clinically relevant therapeutics. The rise of resistant pathogens has led to calls for renewed efforts to identify antimicrobial activities, including expanded screening of streptomycetes. Indeed, it is known that most strains encode >20 secondary metabolites and that many, perhaps most of these, have not been considered for their possible therapeutic use. One roadblock is that many strains do not express their secondary metabolic gene clusters efficiently under laboratory conditions. As one approach to this problem, we have used alleles of a pleiotropic regulator of secondary metabolism from Streptomyces coelicolor to activate secondary biosynthetic gene clusters in heterologous streptomycetes. In one case, we demonstrate the activation of pulvomycin production in S. flavopersicus, a metabolite not previously attributed to this species. We find that the absA1-engineered strains produced sufficient material for purification and characterization. As a result, we identified new, broad-spectrum antimicrobial activities for pulvomycin, including a potent antimicrobial activity against highly antibiotic-resistant Gram-negative and Gram-positive pathogens.

  1. X-37 Flight Demonstrator: A Building Block in NASA's Future Access to Space; X-37 Flight Demonstrator: Orbital Vehicle Technology Development Approach

    NASA Technical Reports Server (NTRS)

    Jacobson, David

    2004-01-01

    Project management issues and subsystems development for the X-37 flight demonstrator are covered in this viewgraph presentation. Subsystems profiled include: thermal protection systems, hot structures, and lithium-ion batteries.

  2. DEVELOPMENT AND DEMONSTRATION OF AN ULTRA LOW NOx COMBUSTOR FOR GAS TURBINES

    SciTech Connect

    NEIL K. MCDOUGALD

    2005-04-30

    Alzeta Corporation has developed surface-stabilized fuel injectors for use with lean premixed combustors which provide extended turndown and ultra-low NOX emission performance. These injectors use a patented technique to form interacting radiant and blue-flame zones immediately above a selectively-perforated porous metal surface. This allows stable operation at low reaction temperatures. This technology is being commercialized under the product name nanoSTAR. Initial tests demonstrated low NOX emissions but, were limited by flashback failure of the injectors. The weld seams required to form cylindrical injectors from flat sheet material were identified as the cause of the failures. The approach for this project was to first develop new fabrication methods to produce injectors without weld seams, verify similar emissions performance to the original flat sheet material and then develop products for microturbines and small gas turbines along parallel development paths. A 37 month project was completed to develop and test a surface stabilized combustion system for gas turbine applications. New fabrication techniques developed removed a technological barrier to the success of the product by elimination of conductive weld seams from the injector surface. The injectors demonstrated ultra low emissions in rig tests conducted under gas turbine operating conditions. The ability for injectors to share a common combustion chamber allowing for deployment in annular combustion liner was also demonstrated. Some further development is required to resolve integration issues related to specific engine constraints, but the nanoSTAR technology has clearly demonstrated its low emissions potential. The overall project conclusions can be summarized: (1) A wet-laid casting method successfully eliminated weld seams from the injector surface without degrading performance. (2) Gas turbine cycle analysis identified several injector designs and control schemes to start and load engines using

  3. DEVELOPMENT AND DEMONSTRATION OF POLYMER MICROENCAPSULATION OF MIXED WASTE USING KINETIC MIXER PROCESSING

    SciTech Connect

    LAGERAAEN,P.R.; KALB,P.D.; MILIAN,L.W.; ADAMS,J.W.

    1997-11-01

    Thermokinetic mixing was investigated as an alternative processing method for polyethylene microencapsulation, a technology well demonstrated for treatment of hazardous, low-level radioactive and low-level mixed wastes. Polyethylene encapsulation by extrusion has been previously shown to be applicable to a wide range of waste types but often pretreatment of the wastes is necessary due to process limitations regarding the maximum waste moisture content and particle size distribution. Development testing was conducted with kinetic mixing in order to demonstrate technology viability and show improved process applicability in these areas. Testing to establish process capabilities and relevant operating parameters was performed with waste surrogates including an aqueous evaporator concentrate and soil. Using a pilot-scale kinetic mixer which was installed and modified for this program, the maximum waste moisture content and particle size was determined. Following process development with surrogate wastes, the technology was successfully demonstrated at BNL using actual mixed waste.

  4. St. Louis' Vocabulary Development Project: A Report on the 1968-69 Demonstration.

    ERIC Educational Resources Information Center

    Tanis, David; And Others

    The Vocabulary Development Project (VDP) was a pilot study designed to demonstrate that significant improvement in language skills is possible with a sustained, massive program of vocabulary exercises. The subjects were inner-city fourth, fifth, and sixth graders. As this project was not a full-blown experimental study, a control…

  5. Development of the Optical Communications Telescope Laboratory: A Laser Communications Relay Demonstration Ground Station

    NASA Technical Reports Server (NTRS)

    Wilson, K. E.; Antsos, D.; Roberts, L. C. Jr.,; Piazzolla, S.; Clare, L. P.; Croonquist, A. P.

    2012-01-01

    The Laser Communications Relay Demonstration (LCRD) project will demonstrate high bandwidth space to ground bi-directional optical communications links between a geosynchronous satellite and two LCRD optical ground stations located in the southwestern United States. The project plans to operate for two years with a possible extension to five. Objectives of the demonstration include the development of operational strategies to prototype optical link and relay services for the next generation tracking and data relay satellites. Key technologies to be demonstrated include adaptive optics to correct for clear air turbulence-induced wave front aberrations on the downlink, and advanced networking concepts for assured and automated data delivery. Expanded link availability will be demonstrated by supporting operations at small sun-Earth-probe angles. Planned optical modulation formats support future concepts of near-Earth satellite user services to a maximum of 1.244 Gb/s differential phase shift keying modulation and pulse position modulations formats for deep space links at data rates up to 311 Mb/s. Atmospheric monitoring instruments that will characterize the optical channel during the link include a sun photometer to measure atmospheric transmittance, a solar scintillometer, and a cloud camera to measure the line of sight cloud cover. This paper describes the planned development of the JPL optical ground station.

  6. A sporulation-specific, sigF-dependent protein, SspA, affects septum positioning in Streptomyces coelicolor

    PubMed Central

    Tzanis, Angelos; Dalton, Kate A; Hesketh, Andrew; den Hengst, Chris D; Buttner, Mark J; Thibessard, Annabelle; Kelemen, Gabriella H

    2014-01-01

    The RNA polymerase sigma factor SigF controls late development during sporulation in the filamentous bacterium Streptomyces coelicolor. The only known SigF-dependent gene identified so far, SCO5321, is found in the biosynthetic cluster encoding spore pigment synthesis. Here we identify the first direct target for SigF, the gene sspA, encoding a sporulation-specific protein. Bioinformatic analysis suggests that SspA is a secreted lipoprotein with two PepSY signature domains. The sspA deletion mutant exhibits irregular sporulation septation and altered spore shape, suggesting that SspA plays a role in septum formation and spore maturation. The fluorescent translational fusion protein SspA–mCherry localized first to septum sites, then subsequently around the surface of the spores. Both SspA protein and sspA transcription are absent from the sigF null mutant. Moreover, in vitro transcription assay confirmed that RNA polymerase holoenzyme containing SigF is sufficient for initiation of transcription from a single sspA promoter. In addition, in vivo and in vitro experiments showed that sspA is a direct target of BldD, which functions to repress sporulation genes, including whiG, ftsZ and ssgB, during vegetative growth, co-ordinating their expression during sporulation septation. PMID:24261854

  7. TRL Assessment of Solar Sail Technology Development Following the 20-Meter System Ground Demonstrator Hardware Testing

    NASA Technical Reports Server (NTRS)

    Young, Roy M.; Adams, Charles L.

    2010-01-01

    The NASA In-Space Propulsion Technology (ISPT) Projects Office sponsored two separate, independent solar sail system design and development demonstration activities during 2002-2005. ATK Space Systems of Goleta, CA was the prime contractor for one development team and L' Garde, Inc. of Tustin, CA was the prime contractor for the other development team. The goal of these activities was to advance the technology readiness level (TRL) of solar sail propulsion from 3 towards 6 by the year 2006. Component and subsystem fabrication and testing were completed successfully, including the ground deployment of 10-meter and 20-meter demonstration hardware systems under vacuum conditions. The deployment and structural testing of the 20-meter solar sail systems was conducted in the 30 meter diameter Space Power Facility thermal-vacuum chamber at NASA Glenn Plum Brook in April though August, 2005. This paper will present the results of the TRL assessment following the solar sail technology development activities associated with the design, development, analysis and testing of the 20-meter system ground demonstrators.

  8. Aging aircraft NDI Development and Demonstration Center (AANC): An overview. [nondestructive inspection

    NASA Technical Reports Server (NTRS)

    Walter, Patrick L.

    1992-01-01

    A major center with emphasis on validation of nondestructive inspection (NDI) techniques for aging aircraft, the Aging Aircraft NDI Development and Demonstration Center (AANC), has been funded by the FAA at Sandia National Laboratories. The Center has been assigned specific tasks in developing techniques for the nondestructive inspection of static engine parts, assessing inspection reliability (POD experiments), developing testbeds for NDI validation, maintaining a FAA library of characterized aircraft structural test specimens, and leasing a hangar to house a high flight cycle transport aircraft for use as a full scale test bed.

  9. In Vivo Analysis of HPr Reveals a Fructose-Specific Phosphotransferase System That Confers High-Affinity Uptake in Streptomyces coelicolor

    PubMed Central

    Nothaft, Harald; Parche, Stephan; Kamionka, Annette; Titgemeyer, Fritz

    2003-01-01

    HPr, the histidine-containing phosphocarrier protein of the bacterial phosphotransferase system (PTS), serves multiple functions in carbohydrate uptake and carbon source regulation in low-G+C-content gram-positive bacteria and in gram-negative bacteria. To assess the role of HPr in the high-G+C-content gram-positive organism Streptomyces coelicolor, the encoding gene, ptsH, was deleted. The ptsH mutant BAP1 was impaired in fructose utilization, while growth on other carbon sources was not affected. Uptake assays revealed that BAP1 could not transport appreciable amounts of fructose, while the wild type showed inducible high-affinity fructose transport with an apparent Km of 2 μM. Complementation and reconstitution experiments demonstrated that HPr is indispensable for a fructose-specific PTS activity. Investigation of the putative fruKA gene locus led to identification of the fructose-specific enzyme II permease encoded by the fruA gene. Synthesis of HPr was not specifically enhanced in fructose-grown cells and occurred also in the presence of non-PTS carbon sources. Transcriptional analysis of ptsH revealed two promoters that are carbon source regulated. In contrast to what happens in other bacteria, glucose repression of glycerol kinase was still operative in a ptsH background, which suggests that HPr is not involved in general carbon regulation. However, fructose repression of glycerol kinase was lost in BAP1, indicating that the fructose-PTS is required for transduction of the signal. This study provides the first molecular genetic evidence of a physiological role of the PTS in S. coelicolor. PMID:12533468

  10. Developing waste disposal options in the underground storage tank - integrated demonstration program

    SciTech Connect

    Cruse, J.M.; Gilchrist, R.L.

    1993-06-01

    The principal objective of the Underground Storage Tank - Integrated Demonstration (UST-ID Program is the demonstration and continued development of technologies suitable for the remediation of USTs. The most promising new technologies from industry, universities, national laboratories, and other government agencies are selected for demonstration, testing, and evaluation. The objective is the eventual transfer of new technologies as part of a system to full-scale remediation at US Department of Energy (DOE) sites and alternately into the private sector. Technologies under development in the UST-ID Program are targeted toward use in remediation actions at the following five DOE participant sites: Hanford, Fernald, Idaho, Oak Ridge, and Savannah River. Combined, these participant sites have more than 300 USTs containing more than 381,800 m{sup 3} (100 Mgal) of high-level and low-level radioactive liquid waste. This paper focuses on the Low-Level Waste Disposal area of the UST-ID, summarizing the two currently funded technology development projects: the Nitrate to Ammonia and Ceramic (NAC) Process and Polyethylene Encapsulation. Both technologies are considered options to the-current baseline disposal approaches being developed at the participant sites. For the Hanford Site, this baseline is a grout waste form that is nearing implementation for disposal of low-level liquid tank wastes.

  11. Mixed Waste Focus Area alternative oxidation technologies development and demonstration program

    SciTech Connect

    Borduin, L.C.; Fewell, T.; Gombert, D.; Priebe, S.

    1998-07-01

    The Mixed Waste Focus Area (MWFA) is currently supporting the development and demonstration of several alternative oxidation technology (AOT) processes for treatment of combustible mixed low-level wastes. The impetus for this support derives from regulatory and political hurdles frequently encountered by traditional thermal techniques, primarily incinerators. AOTs have been defined as technologies that destroy organic material without using open-flame reactions. Whether thermal or nonthermal, the processes have the potential advantages of relatively low-volume gaseous emissions, generation of few or no dioxin/furan compounds, and operation at low enough temperatures that metals (except mercury) and most radionuclides are not volatilized. Technology development and demonstration are needed to confirm and realize the potential of AOTs and to compare them on an equal basis with their fully demonstrated thermal counterparts. AOTs include both thermal and nonthermal processes that oxidize organic wastes but operate under significantly different physical and chemical conditions than incinerators. Nonthermal processes currently being studied include Delphi DETOX and acid digestion at the Savannah River Site, and direct chemical oxidation at Lawrence Livermore National Laboratory. All three technologies are at advanced stages of development or are entering the demonstration phase. Nonflame thermal processes include catalytic chemical oxidation, which is being developed and deployed at Lawrence Berkeley National Laboratory, and team reforming, a commercial process being supported by Department of Energy. Related technologies include two low-flow, secondary oxidation processes (Phoenix and Thermatrix units) that have been tested at MSE, Inc., in Butte, Montana. Although testing is complete on some AOT technologies, most require additional support to complete some or all of the identified development objectives. Brief descriptions, status, and planned paths forward for each

  12. Design, Development,and Testing of Umbillical System Mechanisms for the X-33 Advanced Technology Demonstrator

    NASA Technical Reports Server (NTRS)

    Littlefield, Alan C.; Melton, Gregory S.

    1999-01-01

    The X-33 Advanced Technology Demonstrator is an un-piloted, vertical take-off, horizontal landing spacecraft. The purpose of the X-33 program is to demonstrate technologies that will dramatically lower the cost of access to space. The rocket-powered X-33 will reach an altitude of up to 100 km and speeds between Mach 13 and 15. Fifteen flight tests are planned, beginning in 2000. Some of the key technologies demonstrated will be the linear aerospike engine, improved thermal protection systems, composite fuel tanks and reduced operational timelines. The X-33 vehicle umbilical connections provide monitoring, power, cooling, purge, and fueling capability during horizontal processing and vertical launch operations. Two "rise-ofF' umbilicals for the X-33 have been developed, tested, and installed. The X-33 umbilical systems mechanisms incorporate several unique design features to simplify horizontal operations and provide reliable disconnect during launch.

  13. The VanRS Homologous Two-Component System VnlRSAb of the Glycopeptide Producer Amycolatopsis balhimycina Activates Transcription of the vanHAXSc Genes in Streptomyces coelicolor, but not in A. balhimycina

    PubMed Central

    Kilian, Regina; Frasch, Hans-Joerg; Kulik, Andreas; Wohlleben, Wolfgang

    2016-01-01

    In enterococci and in Streptomyces coelicolor, a glycopeptide nonproducer, the glycopeptide resistance genes vanHAX are colocalized with vanRS. The two-component system (TCS) VanRS activates vanHAX transcription upon sensing the presence of glycopeptides. Amycolatopsis balhimycina, the producer of the vancomycin-like glycopeptide balhimycin, also possesses vanHAXAb genes. The genes for the VanRS-like TCS VnlRSAb, together with the carboxypeptidase gene vanYAb, are part of the balhimycin biosynthetic gene cluster, which is located 2 Mb separate from the vanHAXAb. The deletion of vnlRSAb did not affect glycopeptide resistance or balhimycin production. In the A. balhimycina vnlRAb deletion mutant, the vanHAXAb genes were expressed at the same level as in the wild type, and peptidoglycan (PG) analyses proved the synthesis of resistant PG precursors. Whereas vanHAXAb expression in A. balhimycina does not depend on VnlRAb, a VnlRAb-depending regulation of vanYAb was demonstrated by reverse transcriptase polymerase chain reaction (RT-PCR) and RNA-seq analyses. Although VnlRAb does not regulate the vanHAXAb genes in A. balhimycina, its heterologous expression in the glycopeptide-sensitive S. coelicolor ΔvanRSSc deletion mutant restored glycopeptide resistance. VnlRAb activates the vanHAXSc genes even in the absence of VanS. In addition, expression of vnlRAb increases actinorhodin production and influences morphological differentiation in S. coelicolor. PMID:27420548

  14. An ST-VNS approach for fusion technology development and demonstration

    SciTech Connect

    Cheng, E.T.; Peng, Yueng Kay Martin; Sze, D. K.; Berk, S. E.

    1998-01-01

    A fusion technology development and demonstration concept is conceived using a plasma based volumetric neutron source capable of providing a range of fusion power and wall loading levels during testing, developing, and demonstration stages of operation. The high performance anticipated for the spherical torus (ST) plasma permits large physics margins for the initial neutron wall loading of 0.5 MW/m(2), while still allowing a long-term power reactor relevant physics performance goal of 5 MW/m(2) with reduced margins. An ST-VNS takes advantage of low magnetic field and compact plasma to implement full remote replacement of the center leg of the toroidal field coil, the divertor, the first wall, the shield, and the test blanket modules. At the initial stage of operation, only modest neutron wall loading (0.5 MW/m(2)) would be attempted. Success in achieving reliable operations at 2 MW/m(2) in wall loading at a later stage would fulfill the goals of the VNS; success at 5 MW/m(2) would permit component testing for the demonstration power plants. This approach of ST-VNS provides a realistic opportunity for the ultimate demonstration of safe, reliable, and environmentally attractive fusion systems.

  15. Structural and functional characterization of the alanine racemase from Streptomyces coelicolor A3(2).

    PubMed

    Tassoni, Raffaella; van der Aart, Lizah T; Ubbink, Marcellus; van Wezel, Gilles P; Pannu, Navraj S

    2017-01-29

    The conversion of l-alanine (L-Ala) into d-alanine (D-Ala) in bacteria is performed by pyridoxal phosphate-dependent enzymes called alanine racemases. D-Ala is an essential component of the bacterial peptidoglycan and hence required for survival. The Gram-positive bacterium Streptomyces coelicolor has at least one alanine racemase encoded by alr. Here, we describe an alr deletion mutant of S. coelicolor which depends on D-Ala for growth and shows increased sensitivity to the antibiotic d-cycloserine (DCS). The crystal structure of the alanine racemase (Alr) was solved with and without the inhibitors DCS or propionate, at 1.64 Å and 1.51 Å resolution, respectively. The crystal structures revealed that Alr is a homodimer with residues from both monomers contributing to the active site. The dimeric state of the enzyme in solution was confirmed by gel filtration chromatography, with and without L-Ala or d-cycloserine. The activity of the enzyme was 66 ± 3 U mg(-1) for the racemization of L- to D-Ala, and 104 ± 7 U mg(-1) for the opposite direction. Comparison of Alr from S. coelicolor with orthologous enzymes from other bacteria, including the closely related d-cycloserine-resistant Alr from S. lavendulae, strongly suggests that structural features such as the hinge angle or the surface area between the monomers do not contribute to d-cycloserine resistance, and the molecular basis for resistance therefore remains elusive.

  16. The Iodine Satellite (iSAT) Hall Thruster Demonstration Mission Concept and Development

    NASA Technical Reports Server (NTRS)

    Dankanich, John W.; Polzin, Kurt A.; Calvert, Derek; Kamhawi, Hani

    2014-01-01

    The use of iodine propellant for Hall thrusters has been studied and proposed by multiple organizations due to the potential mission benefits over xenon. In 2013, NASA Marshall Space Flight Center competitively selected a project for the maturation of an iodine flight operational feed system through the Technology Investment Program. Multiple partnerships and collaborations have allowed the team to expand the scope to include additional mission concept development and risk reduction to support a flight system demonstration, the iodine Satellite (iSAT). The iSAT project was initiated and is progressing towards a technology demonstration mission preliminary design review. The current status of the mission concept development and risk reduction efforts in support of this project is presented.

  17. Development and demonstration of the Westinghouse hardware-compressive true time delay (TTD) fiber optic system

    NASA Astrophysics Data System (ADS)

    Goutzoulis, Anastasios P.; Davies, D. Kenneth; Zomp, John M.; Hrycak, Peter; Johnson, Andy

    1994-06-01

    We describe the development and antenna range demonstration of the Westinghouse fiber optic wavelength-multiplexed true time delay steering system for use with broadband phased array antennas. The prototype system is based on a unique hardware compressive architecture and can drive 16 antenna elements over the 0.32 to 2.1 GHz band with 6-bit angular resolution over a +/- 45 degree(s) scan angle.

  18. Crew station research and development facility training for the light helicopter demonstration/validation program

    NASA Technical Reports Server (NTRS)

    Matsumoto, Joy Hamerman; Rogers, Steven; Mccauley, Michael; Salinas, AL

    1992-01-01

    The U.S. Army Crew Station Research and Development Branch (CSRDB) of the Aircraft Simulation Division (AVSCOM) was tasked by the Light Helicopter Program Manager (LH-PM) to provide training to Army personnel in advanced aircraft simulation technology. The purpose of this training was to prepare different groups of pilots to support and evaluate two contractor simulation efforts during the Demonstration/Validation (DEM/VAL) phase of the LH program. The personnel in the CSRDB developed mission oriented training programs to accomplish the objectives, conduct the programs, and provide guidance to army personnel and support personnel throughout the DEM/VAL phase.

  19. The Development and Demonstration of a 360m/10 kA HTS DC Power Cable

    NASA Astrophysics Data System (ADS)

    Xiao, Liye

    With the quick development of renewable energy, it is expected that the electric power from renewable energy would be the dominant one for the future power grid. Due to the specialty of the renewable energy, the HVDC power transmission would be very useful for the transmission of electric power from renewable energy. DC power cable made of High Tc Superconductor (HTS) would be a possible alternative for the construction of HVDC power transmission system. In this chapter, we report the development and demonstration of a 360 m/10 kA HTS DC power cable and the test results.

  20. Vibration isolation technology: An executive summary of systems development and demonstration

    NASA Technical Reports Server (NTRS)

    Grodsinsky, Carlos M.; Logsdon, Kirk A.; Lubomski, Joseph F.

    1993-01-01

    A program was organized to develop the enabling technologies needed for the use of Space Station Freedom as a viable microgravity experimental platform. One of these development programs was the Vibration Isolation Technology (VIT). This technology development program grew because of increased awareness that the acceleration disturbances present on the Space Transportation System (STS) orbiter can and are detrimental to many microgravity experiments proposed for STS, and in the future, Space Station Freedom (SSF). Overall technological organization are covered of the VIT program. Emphasis is given to the results from development and demonstration of enabling technologies to achieve the acceleration requirements perceived as those most likely needed for a variety of microgravity science experiments. In so doing, a brief summary of general theoretical approaches to controlling the acceleration environment of an isolated space based payload and the design and/or performance of two prototype six degree of freedom active magnetic isolation systems is presented.

  1. 1000kW on-site PAFC power plant development and demonstration

    SciTech Connect

    Satomi, Tomohide; Koike, Shunichi; Ishikawa, Ryou

    1996-12-31

    Phosphoric Acid Fuel Cell Technology Research Association (PAFC-TRA) and New Energy and Industrial Technology Development Organization (NEDO) have been conducting a joint project on development of a 5000kW urban energy center type PAFC power plant (pressurized) and a 1000kW on-site PAFC power plant (non-pressurized). The objective of the technical development of 1000kW on-site PAFC power plant is to realize a medium size power plant with an overall efficiency of over 70% and an electrical efficiency of over 36%, that could be installed in a large building as a cogeneration system. The components and system integration development work and the plant design were performed in 1991 and 1992. Manufacturing of the plant and installation at the test site were completed in 1994. PAC test was carried out in 1994, and generation test was started in January 1995. Demonstration test is scheduled for 1995 and 1996.

  2. Microgravity vibration isolation technology: Development to demonstration. Ph.D. Thesis - Case Western Reserve Univ.

    NASA Technical Reports Server (NTRS)

    Grodsinsky, Carlos M.

    1993-01-01

    The low gravity environment provided by space flight has afforded the science community a unique area for the study of fundamental and technological sciences. However, the dynamic environment observed on space shuttle flights and predicted for Space Station Freedom has complicated the analysis of prior 'microgravity' experiments and prompted concern for the viability of proposed space experiments requiring long term, low gravity environments. Thus, isolation systems capable of providing significant improvements to this random environment have been developed. This dissertation deals with the design constraints imposed by acceleration sensitive, microgravity experiment payloads in the unique environment of space. A theoretical background for the inertial feedback and feedforward isolation of a payload was developed giving the basis for two experimental active inertial isolation systems developed for the demonstration of these advanced active isolation techniques. A prototype six degree of freedom digital active isolation system was designed and developed for the ground based testing of an actively isolated payload in three horizontal degrees of freedom. A second functionally equivalent system was built for the multi-dimensional testing of an active inertial isolation system in a reduced gravity environment during low gravity aircraft trajectories. These multi-input multi-output control systems are discussed in detail with estimates on acceleration noise floor performance as well as the actual performance acceleration data. The attenuation performance is also given for both systems demonstrating the advantages between inertial and non-inertial control of a payload for both the ground base environment and the low gravity aircraft acceleration environment. A future goal for this area of research is to validate the technical approaches developed to the 0.01 Hz regime by demonstrating a functional active inertial feedforward/feedback isolation system during orbital flight

  3. TRL Assessment of Solar Sail Technology Development Following the 20-Meter System Ground Demonstrator Hardware Testing

    NASA Technical Reports Server (NTRS)

    Montgomery, Edward E.; Young, Roy M.; Adams, Charles L.

    2007-01-01

    The NASA In-Space Propulsion Technology (ISPT) Projects Office has been sponsoring 2 separate, independent system design and development hardware demonstration activities during 2002-2005. ATK Space Systems of Goleta, CA was the prime contractor for one development team and L'Garde, Inc. of Tustin, CA was the prime contractor for the other development team. The goal of these activities was to advance the technology readiness level (TRL) of solar sail propulsion from 3 towards 6 by the year 2006. Component and subsystem fabrication and testing were completed successfully, including the ground deployment of 10-meter and 20-meter ground demonstration hardware systems under vacuum conditions. The deployment and structural testing of the 20-meter solar sail systems was conducted in the 30 meter diameter Space Power Facility thermal-vacuum chamber at NASA Glenn Plum Brook in April though August, 2005. This paper will present the results of the TRL assessment following the solar sail technology development activities associated with the design, development, analysis and testing of the 20-meter system ground demonstrators. Descriptions of the system designs for both the ATK and L'Garde systems will be presented. Changes, additions and evolution of the system designs will be highlighted. A description of the modeling and analyses activities performed by both teams, as well as testing conducted to raise the TRL of solar sail technology will be presented. A summary of the results of model correlation activities will be presented. Finally, technology gaps identified during the assessment and gap closure plans will be presented, along with "lessons learned", subsequent planning activities and validation flight opportunities for solar sail propulsion technology.

  4. FY94 Office of Technology Development Mixed Waste Operations Robotics Demonstration

    SciTech Connect

    Kriikku, E.M.

    1994-08-30

    The Department of Energy (DOE) Office of Technology Development (OTD) develops technologies to help solve waste management and environmental problems at DOE sites. The OTD includes the Robotics Technology Development Program (RTDP) and the Mixed Waste Integrated Program (MWIP). Together these programs will provide technologies for DOE mixed waste cleanup projects. Mixed waste contains both radioactive and hazardous constituents. DOE sites currently store over 240,000 cubic meters of low level mixed waste and cleanup activities will generate several hundred thousand more cubic meters. Federal and state regulations require that this waste must be processed before final disposal. The OTD RTDP Mixed Waste Operations (MWO) team held several robotic demonstrations at the Savannah River Site (SRS) during November of 1993. Over 330 representatives from DOE, Government Contractors, industry, and universities attended. The MWO team includes: Fernald Environmental Management Project (FEMP), Idaho National Engineering Laboratory (INEL), Lawrence Livermore National Laboratory (LLNL), Oak Ridge National Engineering Laboratory (ORNL), Sandia National Laboratory (SNL), and Savannah River Technology Center (SRTC). SRTC is the lead site for MWO and provides the technical coordinator. The primary demonstration objective was to show that robotic technologies can make DOE waste facilities run better, faster, more cost effective, and safer. To meet the primary objective, the demonstrations successfully showed the following remote waste drum processing activities: non-destructive drum examination, drum transportation, drum opening, removing waste from a drum, characterize and sort waste items, scarify metal waste, and inspect stored drums. To further meet the primary objective, the demonstrations successfully showed the following remote waste box processing activities: swing free crane control, workcell modeling, and torch standoff control.

  5. Affordable Development and Demonstration of a Small NTR Engine and Stage: How Small is Big Enough?

    NASA Technical Reports Server (NTRS)

    Borowski, S. K.; Sefcik, R. J.; Fittje, J. E.; McCurdy, D. R.; Qualls, A. L.; Schnitzler, B. G.; Werner, J.; Weitzberg, A.; Joyner, C. R.

    2015-01-01

    In FY11, NASA formulated a plan for Nuclear Thermal Propulsion (NTP) development that included Foundational Technology Development followed by system-level Technology Demonstrations The ongoing NTP project, funded by NASAs Advanced Exploration Systems (AES) program, is focused on Foundational Technology Development and includes 5 key task activities:(1) Fuel element fabrication and non-nuclear validation testing of heritage fuel options;(2) Engine conceptual design;(3) Mission analysis and engine requirements definition;(4) Identification of affordable options for ground testing; and(5) Formulation of an affordable and sustainable NTP development program Performance parameters for Point of Departure designs for a small criticality-limited and full size 25 klbf-class engine were developed during FYs 13-14 using heritage fuel element designs for both RoverNERVA Graphite Composite (GC) and Ceramic Metal (Cermet) fuel forms To focus the fuel development effort and maximize use of its resources, the AES program decided, in FY14, that a leader-follower down selection between GC and cermet fuel was required An Independent Review Panel (IRP) was convened by NASA and tasked with reviewing the available fuel data and making a recommendation to NASA. In February 2015, the IRP recommended and the AES program endorsed GC as the leader fuel In FY14, a preliminary development schedule DDTE plan was produced by GRC, DOE industry for the AES program. Assumptions, considerations and key task activities are presented here Two small (7.5 and 16.5 klbf) engine sizes were considered for ground and flight technology demonstration within a 10-year timeframe; their ability to support future human exploration missions was also examined and a recommendation on a preferred size is provided.

  6. Development and Demonstration of Sustainable Surface Infrastructure for Moon/Mars Exploration

    NASA Technical Reports Server (NTRS)

    Sanders, Gerald B.; Larson, William E.; Picard, Martin

    2011-01-01

    For long-term human exploration of the Moon and Mars to be practical, affordable, and sustainable, future missions must be able to identify and utilize resources at the site of exploration. The ability to characterize, extract, processes, and separate products from local material, known as In-Situ Resource Utilization (ISRU), can provide significant reductions in launch mass, logistics, and development costs while reducing risk through increased mission flexibility and protection as well as increased mission capabilities in the areas of power and transportation. Making mission critical consumables like propellants, fuel cell reagents and life support gases, as well as in-situ crew/hardware protection and energy storage capabilities can significantly enhance robotic and human science and exploration missions, however other mission systems need to be designed to interface with and utilize these in-situ developed products and services from the start or the benefits will be minimized or eliminated. This requires a level of surface and transportation system development coordination not typically utilized during early technology and system development activities. An approach being utilized by the US National Aeronautics and Space Administration and the Canadian Space Agency has been to utilize joint analogue field demonstrations to focus technology development activities to demonstrate and integrate new and potentially game changing. mission critical capabilities that would enable an affordable and sustainable surface infrastructure for lunar and Mars robotic and human exploration. Two analogue field tests performed in November 2008 and February 2010 demonstrated first generation capabilities for lunar resource prospecting, exploration site preparation, and oxygen extraction from regolith while initiating integration with mobility, science, fuel cell power, and propulsion disciplines. A third analogue field test currently planned for June 2012 will continue and expand

  7. Technical support to the Solvent Refined Coal (SRC) demonstration projects: assessment of current research and development

    SciTech Connect

    Edwards, M.S.; Rodgers, B.R.; Brown, C.H.; Carlson, P.K.; Gambill, W.R.; Gilliam, T.M.; Holmes, J.M.; Krishnan, R.P.; Parsly, L.F.

    1980-12-01

    A program to demonstrate Solvent Refined Coal (SRC) technology has been initiated by the US Department of Energy (DOE) in partnership with two industrial groups. Project management responsibility has been assigned to the Oak Ridge Operations Office (ORO) of DOE. ORO requested that the Oak Ridge National Laboratory assess current research and development (R and D) activities and develop recommendations for those activities that might contribute to successful completion of the SRC demonstration plant projects. The objectives of this final report are to discuss in detail the problem areas in SRC; to discuss the current and planned R and D investigations relevant to the problems identified; and to suggest appropriate R and D activities in support of designs for the SRC demonstration plants. Four types of R and D activities are suggested: continuation of present and planned activities; coordination of activities and results, present and proposed; extension/redirection of activities not involving major equipment purchase or modifications; and new activities. Important examples of the first type of activity include continuation of fired heater, slurry rheology, and slurry mixing studies at Ft. Lewis. Among the second type of activity, coordination of data acquisition and interpretation is recommended in the areas of heat transfer, vapor/liquid equilibria, and physical properties. Principal examples of recommendations for extension/redirection include screening studies at laboratory scale on the use of carbonaceous precoat (e.g., anthracite) infiltration, and 15- to 30-day continuous tests of the Texaco gasifier at the Texaco Montebello facility (using SRC residues).

  8. Development and Evaluation of the Habitat Demonstration Unit Medical Operations Workstation and Opportunities for Future Research

    NASA Technical Reports Server (NTRS)

    Howard, Robert L., Jr.

    2012-01-01

    As NASA develops missions to leave Earth orbit and explore distant destinations (Mars, Moon, Asteroids) it is necessary to rethink human spaceflight paradigms in the life sciences. Standards developed for low earth orbit human spaceflight may not be fully applicable and in-space research may be required to develop new standards. Preventative and emergency medical care may require new capabilities never before used in space. Due to spacecraft volume limitations, this work area may also be shared with various animal and plant life science research. This paper explores the prototype Medical Operations Workstation within the NASA Habitat Demonstration Unit and discusses some of the lessons learned from field analogue missions involving the workstation. Keywords: Exploration, medical, health, crew, injury emergency, biology, animal, plant, science, preventative, emergency.

  9. Development and demonstration of a flutter-suppression system using active controls. [wind tunnel tests

    NASA Technical Reports Server (NTRS)

    Sandford, M. C.; Abel, I.; Gray, D. L.

    1975-01-01

    The application of active control technology to suppress flutter was demonstrated successfully in the transonic dynamics tunnel with a delta-wing model. The model was a simplified version of a proposed supersonic transport wing design. An active flutter suppression method based on an aerodynamic energy criterion was verified by using three different control laws. The first two control laws utilized both leading-edge and trailing-edge active control surfaces, whereas the third control law required only a single trailing-edge active control surface. At a Mach number of 0.9 the experimental results demonstrated increases in the flutter dynamic pressure from 12.5 percent to 30 percent with active controls. Analytical methods were developed to predict both open-loop and closed-loop stability, and the results agreed reasonably well with the experimental results.

  10. Water Use Optimization Toolset Project: Development and Demonstration Phase Draft Report

    SciTech Connect

    Gasper, John R.; Veselka, Thomas D.; Mahalik, Matthew R.; Hayse, John W.; Saha, Samrat; Wigmosta, Mark S.; Voisin, Nathalie; Rakowski, Cynthia; Coleman, Andre; Lowry, Thomas S.

    2014-05-19

    This report summarizes the results of the development and demonstration phase of the Water Use Optimization Toolset (WUOT) project. It identifies the objective and goals that guided the project, as well as demonstrating potential benefits that could be obtained by applying the WUOT in different geo-hydrologic systems across the United States. A major challenge facing conventional hydropower plants is to operate more efficiently while dealing with an increasingly uncertain water-constrained environment and complex electricity markets. The goal of this 3-year WUOT project, which is funded by the U.S. Department of Energy (DOE), is to improve water management, resulting in more energy, revenues, and grid services from available water, and to enhance environmental benefits from improved hydropower operations and planning while maintaining institutional water delivery requirements. The long-term goal is for the WUOT to be used by environmental analysts and deployed by hydropower schedulers and operators to assist in market, dispatch, and operational decisions.

  11. NASA Systems Autonomy Demonstration Project - Development of Space Station automation technology

    NASA Technical Reports Server (NTRS)

    Bull, John S.; Brown, Richard; Friedland, Peter; Wong, Carla M.; Bates, William

    1987-01-01

    A 1984 Congressional expansion of the 1958 National Aeronautics and Space Act mandated that NASA conduct programs, as part of the Space Station program, which will yield the U.S. material benefits, particularly in the areas of advanced automation and robotics systems. Demonstration programs are scheduled for automated systems such as the thermal control, expert system coordination of Station subsystems, and automation of multiple subsystems. The programs focus the R&D efforts and provide a gateway for transfer of technology to industry. The NASA Office of Aeronautics and Space Technology is responsible for directing, funding and evaluating the Systems Autonomy Demonstration Project, which will include simulated interactions between novice personnel and astronauts and several automated, expert subsystems to explore the effectiveness of the man-machine interface being developed. Features and progress on the TEXSYS prototype thermal control system expert system are outlined.

  12. Development of LM10-MIRA LOX/LNG expander cycle demonstrator engine

    NASA Astrophysics Data System (ADS)

    Rudnykh, Mikhail; Carapellese, Stefano; Liuzzi, Daniele; Arione, Luigi; Caggiano, Giuseppe; Bellomi, Paolo; D'Aversa, Emanuela; Pellegrini, Rocco; Lobov, S. D.; Gurtovoy, A. A.; Rachuk, V. S.

    2016-09-01

    This article contains results of joint works by Konstruktorskoe Buro Khimavtomatiki (KBKhA, Russia) and AVIO Company (Italy) on creation of the LM10-MIRA liquid-propellant rocket demonstrator engine for the third stage of the upgraded "Vega" launcher.Scientific and research activities conducted by KBKhA and AVIO in 2007-2014 in the frame of the LYRA Program, funded by the Italian Space Agency, with ELV as Prime contractor, and under dedicated ASI-Roscosmos inter-agencies agreement, were aimed at development and testing of a 7.5 t thrust expander cycle demonstrator engine propelled by oxygen and liquid natural gas (further referred to as LNG).

  13. Lipoprotein N-acyl transferase (Lnt1) is dispensable for protein O-mannosylation by Streptomyces coelicolor.

    PubMed

    Córdova-Dávalos, Laura Elena; Espitia, Clara; González-Cerón, Gabriela; Arreguín-Espinosa, Roberto; Soberón-Chávez, Gloria; Servín-González, Luis

    2014-01-01

    A protein glycosylation system related to that for protein mannosylation in yeast is present in many actinomycetes. This system involves polyprenyl phosphate mannose synthase (Ppm), protein mannosyl transferase (Pmt), and lipoprotein N-acyl transferase (Lnt). In this study, we obtained a series of mutants in the ppm (sco1423), lnt1 (sco1014), and pmt (sco3154) genes of Streptomyces coelicolor, which encode Ppm, Lnt1, and Pmt, to analyze their requirement for glycosylation of the heterologously expressed Apa glycoprotein of Mycobacterium tuberculosis. The results show that both Ppm and Pmt were required for Apa glycosylation, but that Lnt1 was dispensable for both Apa and the bacteriophage φC31 receptor glycosylation. A bacterial two-hybrid assay revealed that contrary to M. tuberculosis, Lnt1 of S. coelicolor does not interact with Ppm. The D2 catalytic domain of M. tuberculosisPpm was sufficient for complementation of an S. coelicolor double mutant lacking Lnt1 and Ppm, both for Apa glycosylation and for glycosylation of φC31 receptor. On the other hand, M. tuberculosisPmt was not active in S. coelicolor, even when correctly localized to the cytoplasmic membrane, showing fundamental differences in the requirements for Pmt activity in these two species.

  14. Microgrid Design, Development and Demonstration - Final Report for Phase I and Phase II

    SciTech Connect

    Bose, Sumit; Krok, Michael

    2011-02-08

    This document constitutes GE’s final report for the Microgrid Design, Development and Demonstration program for DOE’s Office of Electricity Delivery and Energy Reliability, Award DE-FC02-05CH11349. It contains the final report for Phase I in Appendix I, and the results the work performed in Phase II. The program goal was to develop and demonstrate a Microgrid Energy Management (MEM) framework for a broad set of Microgrid applications that provides unified controls, protection, and energy management. This project contributed to the achievement of the U.S. Department of Energy’s Renewable and Distributed Systems Integration Program goals by developing a fully automated power delivery microgrid network that: - Reduces carbon emissions and emissions of other air pollutants through increased use of optimally dispatched renewable energy, - Increases asset use through integration of distributed systems, - Enhances reliability, security, and resiliency from microgrid applications in critical infrastructure protection, constrained areas of the electric grid, etc. - Improves system efficiency with on-site, distributed generation and improved economic efficiency through demand-side management.

  15. Northwest Hazardous Waste Research, Development, and Demonstration Center: Program Plan. [Contains glossary

    SciTech Connect

    Not Available

    1988-02-01

    The Northwest Hazardous Waste Research, Development, and Demonstration Center was created as part of an ongoing federal effort to provide technologies and methods that protect human health and welfare and environment from hazardous wastes. The Center was established by the Superfund Amendments and Reauthorization Act (SARA) to develop and adapt innovative technologies and methods for assessing the impacts of and remediating inactive hazardous and radioactive mixed-waste sites. The Superfund legislation authorized $10 million for Pacific Northwest Laboratory to establish and operate the Center over a 5-year period. Under this legislation, Congress authorized $10 million each to support research, development, and demonstration (RD and D) on hazardous and radioactive mixed-waste problems in Idaho, Montana, Oregon, and Washington, including the Hanford Site. In 1987, the Center initiated its RD and D activities and prepared this Program Plan that presents the framework within which the Center will carry out its mission. Section 1.0 describes the Center, its mission, objectives, organization, and relationship to other programs. Section 2.0 describes the Center's RD and D strategy and contains the RD and D objectives, priorities, and process to be used to select specific projects. Section 3.0 contains the Center's FY 1988 operating plan and describes the specific RD and D projects to be carried out and their budgets and schedules. 9 refs., 18 figs., 5 tabs.

  16. The development and testing of the Lens Antenna Deployment Demonstration (LADD) test article

    NASA Technical Reports Server (NTRS)

    Pugh, Mark L.; Denton, Robert J., Jr.; Strange, Timothy J.

    1993-01-01

    The USAF Rome Laboratory and NASA Marshall Space Flight Center, through contract to Grumman Corporation, have developed a space-qualifiable test article for the Strategic Defense Initiative Organization to demonstrate the critical structural and mechanical elements of single-axis roll-out membrane deployment for Space Based Radar (SBR) applications. The Lens Antenna Deployment Demonstration (LADD) test article, originally designed as a shuttle-attached flight experiment, is a large precision space structure which is representative of operational designs for space-fed lens antennas. Although the flight experiment was cancelled due to funding constraints and major revisions in the Strategic Defense System (SDS) architecture, development of this test article was completed in June 1989. To take full advantage of the existence of this unique structure, a series of ground tests are proposed which include static, dynamic, and thermal measurements in a simulated space environment. An equally important objective of these tests is the verification of the analytical tools used to design and develop large precision space structures.

  17. Energy Savings Potential and Research, Development, & Demonstration Opportunities for Commercial Building Appliances

    SciTech Connect

    Zogg, Robert; Goetzler, William; Ahlfeldt, Christopher; Hiraiwa, Hirokazu; Sathe, Amul; Sutherland, Timothy

    2009-12-01

    This study characterizes and assesses the appliances used in commercial buildings. The primary objectives of this study were to document the energy consumed by commercial appliances and identify research, development and demonstration (RD&D) opportunities for efficiency improvements, excluding product categories such as HVAC, building lighting, refrigeration equipment, and distributed generation systems. The study included equipment descriptions, characteristics of the equipment’s market, national energy consumption, estimates of technical potential for energy-saving technologies, and recommendations for U.S. Department of Energy programs that can promote energy savings in commercial appliances.

  18. Development and Comparison of Mechanical Structures for FNAL 15 T Nb$_3$Sn Dipole Demonstrator

    SciTech Connect

    Novitski, I.; Zlobin, A. V.

    2016-11-08

    Main design challenges for 15 T accelerator magnets are large Lorentz forces at this field level. The large Lorentz forces generate high stresses in the coil and mechanical structure and, thus, need stress control to maintain them at the acceptable level for brittle Nb3Sn coils and other elements of magnet mechanical structure. To provide these conditions and achieve the design field in the FNAL 15 T dipole demonstrator, several mechanical structures have been developed and analysed. The possibilities and limitations of these designs are discussed in this paper

  19. Disruption of Macrodomain Protein SCO6735 Increases Antibiotic Production in Streptomyces coelicolor*

    PubMed Central

    Lalić, Jasna; Posavec Marjanović, Melanija; Palazzo, Luca; Perina, Dragutin; Sabljić, Igor; Žaja, Roko; Colby, Thomas; Pleše, Bruna; Halasz, Mirna; Jankevicius, Gytis; Bucca, Giselda; Ahel, Marijan; Matić, Ivan; Ćetković, Helena; Luić, Marija; Mikoč, Andreja; Ahel, Ivan

    2016-01-01

    ADP-ribosylation is a post-translational modification that can alter the physical and chemical properties of target proteins and that controls many important cellular processes. Macrodomains are evolutionarily conserved structural domains that bind ADP-ribose derivatives and are found in proteins with diverse cellular functions. Some proteins from the macrodomain family can hydrolyze ADP-ribosylated substrates and therefore reverse this post-translational modification. Bacteria and Streptomyces, in particular, are known to utilize protein ADP-ribosylation, yet very little is known about their enzymes that synthesize and remove this modification. We have determined the crystal structure and characterized, both biochemically and functionally, the macrodomain protein SCO6735 from Streptomyces coelicolor. This protein is a member of an uncharacterized subfamily of macrodomain proteins. Its crystal structure revealed a highly conserved macrodomain fold. We showed that SCO6735 possesses the ability to hydrolyze PARP-dependent protein ADP-ribosylation. Furthermore, we showed that expression of this protein is induced upon DNA damage and that deletion of this protein in S. coelicolor increases antibiotic production. Our results provide the first insights into the molecular basis of its action and impact on Streptomyces metabolism. PMID:27634042

  20. The Streptomyces coelicolor Developmental Transcription Factor σBldN Is Synthesized as a Proprotein

    PubMed Central

    Bibb, Maureen J.; Buttner, Mark J.

    2003-01-01

    bldN is one of a set of genes required for the formation of specialized, spore-bearing aerial hyphae during differentiation in the mycelial bacterium Streptomyces coelicolor. Previous analysis (M. J. Bibb et al., J. Bacteriol. 182:4606-4616, 2000) showed that bldN encodes a member of the extracytoplasmic function subfamily of RNA polymerase σ factors and that translation from the most strongly predicted start codon (GTG1) would give rise to a σ factor having an unusual N-terminal extension of ca. 86 residues. Here, by using a combination of site-directed mutagenesis and immunoblot analysis, we provide evidence that all bldN translation arises from initiation at GTG1 and that the primary translation product is a proprotein (pro-σBldN) that is proteolytically processed to a mature species (σBldN) by removal of most of the unusual N-terminal extension. A time course taken during differentiation of the wild type on solid medium showed early production of pro-σBldN and the subsequent appearance of mature σBldN, which was concomitant with aerial mycelium formation and the disappearance of pro-σBldN. Two genes encoding members of a family of metalloproteases that are involved in the regulated proteolytic processing of transcription factors in other organisms were identified in the S. coelicolor genome, but their disruption did not affect differentiation or pro-σBldN processing. PMID:12644505

  1. Engineering Streptomyces coelicolor for heterologous expression of secondary metabolite gene clusters

    PubMed Central

    Gomez‐Escribano, Juan Pablo; Bibb, Mervyn J.

    2011-01-01

    Summary We have constructed derivatives of Streptomyces coelicolor M145 as hosts for the heterologous expression of secondary metabolite gene clusters. To remove potentially competitive sinks of carbon and nitrogen, and to provide a host devoid of antibiotic activity, we deleted four endogenous secondary metabolite gene clusters from S. coelicolor M145 – those for actinorhodin, prodiginine, CPK and CDA biosynthesis. We then introduced point mutations into rpoB and rpsL to pleiotropically increase the level of secondary metabolite production. Introduction of the native actinorhodin gene cluster and of gene clusters for the heterologous production of chloramphenicol and congocidine revealed dramatic increases in antibiotic production compared with the parental strain. In addition to lacking antibacterial activity, the engineered strains possess relatively simple extracellular metabolite profiles. When combined with liquid chromatography and mass spectrometry, we believe that these genetically engineered strains will markedly facilitate the discovery of new compounds by heterologous expression of cloned gene clusters, particularly the numerous cryptic secondary metabolic gene clusters that are prevalent within actinomycete genome sequences. PMID:21342466

  2. Global features of gene expression on the proteome and transcriptome levels in S. coelicolor during germination.

    PubMed

    Strakova, Eva; Bobek, Jan; Zikova, Alice; Vohradsky, Jiri

    2013-01-01

    Streptomycetes have been studied mostly as producers of secondary metabolites, while the transition from dormant spores to an exponentially growing culture has largely been ignored. Here, we focus on a comparative analysis of fluorescently and radioactively labeled proteome and microarray acquired transcriptome expressed during the germination of Streptomyces coelicolor. The time-dynamics is considered, starting from dormant spores through 5.5 hours of growth with 13 time points. Time series of the gene expressions were analyzed using correlation, principal components analysis and an analysis of coding genes utilization. Principal component analysis was used to identify principal kinetic trends in gene expression and the corresponding genes driving S. coelicolor germination. In contrast with the correlation analysis, global trends in the gene/protein expression reflected by the first principal components showed that the prominent patterns in both the protein and the mRNA domains are surprisingly well correlated. Analysis of the number of expressed genes identified functional groups activated during different time intervals of the germination.

  3. Carbon Catabolite Regulation of Secondary Metabolite Formation and Morphological Differentiation in Streptomyces coelicolor.

    PubMed

    Romero-Rodríguez, A; Ruiz-Villafán, B; Tierrafría, V H; Rodríguez-Sanoja, R; Sánchez, S

    2016-11-01

    In the genus Streptomyces, carbon utilization is of significant importance for the expression of genes involved in morphological differentiation and antibiotic production. However, there is little information about the mechanism involved in these effects. In the present work, it was found that glucose exerted a suppressive effect on the Streptomyces coelicolor actinorhodin (Act) and undecylprodigiosin (Red) production, as well as in its morphological differentiation. Accordingly, using a high-density microarray approach in S. coelicolor grown under glucose repression, at early growth stages, a negative effect was exerted on the transcription of genes involved in Act and Red production, when compared with non-repressive conditions. Seven genes of Act and at least ten genes of Red production were down-regulated by glucose. Stronger repression was observed on the initial steps of antibiotics formation. On the contrary, the coelimycin P1 cluster was up-regulated by glucose. Regarding differentiation, no sporulation was observed in the presence of glucose and expression of a set of genes of the bld cascade was repressed as well as chaplins and rodlins genes. Finally, a series of transcriptional regulators involved in both processes were up- or down-regulated by glucose. This is the first global transcriptomic approach performed to understand the molecular basis of the glucose effect on the synthesis of secondary metabolism and differentiation in the genus Streptomyces. The results of this study are opening new avenues for further exploration.

  4. Engineering development and demonstration of DETOX{sup SM} wet oxidation for mixed waste treatment

    SciTech Connect

    Dhooge, P.M.; Goldblatt, S.D.; Moslander, J.E.; Robertson, D.T.; Rogers, T.W.; Zigmond, J.A.

    1997-12-01

    DETOX{sup SM}, a catalyzed chemical oxidation process, is under development for treatment of hazardous and mixed wastes at Department of Energy sites. To support this effort, developmental engineering studies have been formed for aspects of the process to help ensure safe and effective operation. Subscale agitation studies have been preformed to identify a suitable mixing head and speed for the primary reaction vessel agitator. Mechanisms for feeding solid waste materials to the primary reaction vessel have been investigated. Filtration to remove solid field process residue, and the use of various filtration aids, has been studied. Extended compatibility studies on the materials of construction have been performed. Due to a change to Rocky Flats Environmental Technology Site (RFETS) for the mixed waste portion of the demonstration, types of wastes suitable and appropriate for treatment at RFETS had to be chosen. A Prototype unit has been fabricated and will be demonstrated on hazardous and mixed wastes at Savannah River Site (SRS) and RFETS during 1997 and 1998. The unit is in shakedown testing at present. Data validation and an engineering evaluation will be performed during the demonstration.

  5. Development of the first demonstration CFB boiler for gas and steam cogeneration

    SciTech Connect

    Fang, M; Luo, Z.; Li, X.; Wang, Q.; Shi, Z.; Ni, M.; Cen, K.

    1997-12-31

    To solve the shortage of gas and steam supply in the small towns of the country, a new gas steam cogeneration system has been developed. On the basis of the fundamental research on the system, a demonstration gas steam cogeneration system has been designed. As the phase 1 of the project, a 75t/h demonstration CFB boiler for gas steam cogeneration has been erected and operated at Yangzhong Thermal Power Plant of China. This paper introduces the first 75t/h demonstration CFB boiler for gas steam cogeneration. Due to the need of gas steam cogeneration process, the boiler has the features of high temperature cyclone separation, high solid recycle ratio, staged combustion and an external heat exchanger adjusting bed temperature and heat load. The operation results show that the boiler has wide fuel adaptability and the heating value of the coal changes from 14MJ/Kg to 25MJ/Kg. The heat load changes from 85t/h to 28t/h while steam parameter is maintained at the normal conditions. The combustion efficiency of the boiler attain 98%. The boiler design and operation experiences may be a guide to the design and operation of larger CFB units in the future.

  6. Development of a 15 T Nb3Sn accelerator dipole demonstrator at Fermilab

    DOE PAGES

    Novitski, I.; Andreev, N.; Barzi, E.; ...

    2016-06-01

    Here, a 100 TeV scale Hadron Collider (HC) with a nominal operation field of at least 15 T is being considered for the post-LHC era, which requires using the Nb3Sn technology. Practical demonstration of this field level in an accelerator-quality magnet and substantial reduction of the magnet costs are the key conditions for realization of such a machine. FNAL has started the development of a 15 T Nb3Sn dipole demonstrator for a 100 TeV scale HC. The magnet design is based on 4-layer shell type coils, graded between the inner and outer layers to maximize the performance and reduce themore » cost. The experience gained during the Nb3Sn magnet R&D is applied to different aspects of the magnet design. This paper describes the magnetic and structural designs and parameters of the 15 T Nb3Sn dipole and the steps towards the demonstration model fabrication.« less

  7. Office of Technology Development`s Research, Development, Demonstration, Testing and Evaluation Mid-Year Program Review. Volume 1

    SciTech Connect

    Not Available

    1994-08-01

    This document presents brief summaries of waste management, remedial action, decommissioning/decontamination, and waste processing programs and issues currently being developed at Department of Energy Facilities.

  8. A Summary fo Solar Sail Technology Developments and Proposed Demonstration Missions

    NASA Technical Reports Server (NTRS)

    Garner, Charles; Diedrich, Benjamin; Leipold, Manfred

    1999-01-01

    NASA's drive to reduce mission costs and accept the risk of incorporating innovative, high payoff technologies into it's missions while simultaneously undertaking ever more difficult missions has sparked a greatly renewed interest in solar sails. From virtually no technology or flight mission studies activity three years ago solar sails are now included in NOAA, NASA, DOD, DLR, ESA and ESTEC technology development programs and technology roadmaps. NASA programs include activities at Langley Research Center, Jet Propulsion Laboratory, Marshall Space Flight Center, Goddard Space Flight Center, and the NASA Institute for Advanced Concepts; NOAA has received funding for a proposed solar sail mission; DLR is designing and fabricating a 20-m laboratory model sail, there are four demonstration missions under study at industry, NASA, DOD and Europe, two new text books on solar sailing were recently published and one new test book is planned. This paper summarizes these on-going developments in solar sails.

  9. Waste management technology development and demonstration programs at Brookhaven National Laboratory

    NASA Technical Reports Server (NTRS)

    Kalb, Paul D.; Colombo, Peter

    1991-01-01

    Two thermoplastic processes for improved treatment of radioactive, hazardous, and mixed wastes were developed from bench scale through technology demonstration: polyethylene encapsulation and modified sulfur cement encapsulation. The steps required to bring technologies from the research and development stage through full scale implementation are described. Both systems result in durable waste forms that meet current Nuclear Regulatory Commission and Environmental Protection Agency regulatory criteria and provide significant improvements over conventional solidification systems such as hydraulic cement. For example, the polyethylene process can encapsulate up to 70 wt pct. nitrate salt, compared with a maximum of about 20 wt pct. for the best hydraulic cement formulation. Modified sulfur cement waste forms containing as much as 43 wt pct. incinerator fly ash were formulated, whereas the maximum quantity of this waste in hydraulic cement is 16 wt pct.

  10. Initial Development and Pilot Study Design of Interactive Lecture Demonstrations for ASTRO 101

    NASA Astrophysics Data System (ADS)

    Schwortz, Andria C.; French, D. A; Gutierrez, Joseph V; Sanchez, Richard L; Slater, Timothy F.; Tatge, Coty

    2014-06-01

    Interactive lecture demonstrations (ILDs) have repeatedly shown to be effective tools for improving student achievement in the context of learning physics. As a first step toward systematic development of interactive lecture demonstrations in ASTRO 101, the introductory astronomy survey course, a systematic review of education research, describing educational computer simulations (ECSs) reveals that initial development requires a targeted study of how ASTRO 101 students respond to ECSs in the non-science majoring undergraduate lecture setting. In this project we have adopted the process by which ILDs were designed, pilot-tested, and successfully implemented in the context of physics teaching (Sokoloff & Thornton, 1997; Sokoloff & Thornton, 2004). We have designed the initial pilot-test set of ASTRO 101 ILD instructional materials relying heavily on ECSs. Both an instructor’s manual and a preliminary classroom-ready student workbook have been developed, and we are implementing a pilot study to explore their effectiveness in communicating scientific content, and the extent to which they might enhance students’ knowledge of and perception about astronomy and science in general. The study design uses a pre-/post-test quasi-experimental study design measuring students’ normalized gain scores, calculated as per Hake (1998) and Prather (2009), using a slightly modified version of S. Slater’s (2011) Test Of Astronomy STandards TOAST combined with other instruments. The results of this initial study will guide the iterative development of ASTRO 101 ILDs that are intended to both be effective at enhancing student achievement and easy for instructors to successfully implement.

  11. Identification of a Gene Negatively Affecting Antibiotic Production and Morphological Differentiation in Streptomyces coelicolor A3(2)▿

    PubMed Central

    Li, Wencheng; Ying, Xin; Guo, Yuzheng; Yu, Zhen; Zhou, Xiufen; Deng, Zixin; Kieser, Helen; Chater, Keith F.; Tao, Meifeng

    2006-01-01

    SC7A1 is a cosmid with an insert of chromosomal DNA from Streptomyces coelicolor A3(2). Its insertion into the chromosome of S. coelicolor strains caused a duplication of a segment of ca. 40 kb and delayed actinorhodin antibiotic production and sporulation, implying that SC7A1 carried a gene negatively affecting these processes. The subcloning of SC7A1 insert DNA resulted in the identification of the open reading frame SCO5582 as nsdA, a gene negatively affecting Streptomyces differentiation. The disruption of chromosomal nsdA caused the overproduction of spores and of three of four known S. coelicolor antibiotics of quite different chemical types. In at least one case (that of actinorhodin), this was correlated with premature expression of a pathway-specific regulatory gene (actII-orf4), implying that nsdA in the wild-type strain indirectly repressed the expression of the actinorhodin biosynthesis cluster. nsdA expression was up-regulated upon aerial mycelium initiation and was strongest in the aerial mycelium. NsdA has DUF921, a Streptomyces protein domain of unknown function and a conserved SXR site. A site-directed mutation (S458A) in this site in NsdA abolished its function. Blast searching showed that NsdA homologues are present in some Streptomyces genomes. Outside of streptomycetes, NsdA-like proteins have been found in several actinomycetes. The disruption of the nsdA-like gene SCO4114 had no obvious phenotypic effects on S. coelicolor. The nsdA orthologue SAV2652 in S. avermitilis could complement the S. coelicolor nsdA-null mutant phenotype. PMID:17041057

  12. Two-Phase Flow Technology Developed and Demonstrated for the Vision for Exploration

    NASA Technical Reports Server (NTRS)

    Sankovic, John M.; McQuillen, John B.; Lekan, Jack F.

    2005-01-01

    NASA s vision for exploration will once again expand the bounds of human presence in the universe with planned missions to the Moon and Mars. To attain the numerous goals of this vision, NASA will need to develop technologies in several areas, including advanced power-generation and thermal-control systems for spacecraft and life support. The development of these systems will have to be demonstrated prior to implementation to ensure safe and reliable operation in reduced-gravity environments. The Two-Phase Flow Facility (T(PHI) FFy) Project will provide the path to these enabling technologies for critical multiphase fluid products. The safety and reliability of future systems will be enhanced by addressing focused microgravity fluid physics issues associated with flow boiling, condensation, phase separation, and system stability, all of which are essential to exploration technology. The project--a multiyear effort initiated in 2004--will include concept development, normal-gravity testing (laboratories), reduced gravity aircraft flight campaigns (NASA s KC-135 and C-9 aircraft), space-flight experimentation (International Space Station), and model development. This project will be implemented by a team from the NASA Glenn Research Center, QSS Group, Inc., ZIN Technologies, Inc., and the Extramural Strategic Research Team composed of experts from academia.

  13. Nevada Test Site-Directed Research, Development, and Demonstration. FY2005 report

    SciTech Connect

    Lewis, Will

    2006-09-01

    The Nevada Test Site-Directed Research, Development, and Demonstration (SDRD) program completed a very successful year of research and development activities in FY 2005. Fifty new projects were selected for funding this year, and five FY 2004 projects were brought to conclusion. The total funds expended by the SDRD program were $5.4 million, for an average per project cost of just under $100,000. Two external audits of SDRD accounting practices were conducted in FY 2005. Both audits found the program's accounting practices consistent with the requirements of DOE Order 413.2A, and one included the observation that the NTS contractor ''did an exceptional job in planning and executing year-start activities.'' Highlights for the year included: the filing of 18 invention disclosures for intellectual property generated by FY 2005 projects; programmatic adoption of 17 FY 2004 SDRD-developed technologies; participation in the tri-lab Laboratory Directed Research and Development (LDRD) and SDRD program review that was broadly attended by NTS, NNSA, LDRD, and U.S. Department of Homeland Security representatives; peer reviews of all FY 2005 projects; and the successful completion of 55 R&D projects, as presented in this report.

  14. Research, Development and Demonstration of Micro-CHP System for Residential Applications

    SciTech Connect

    Karl Mayer

    2010-03-31

    ECR International and its joint venture company, Climate Energy, are at the forefront of the effort to deliver residential-scale combined heat and power (Micro-CHP) products to the USA market. Part of this substantial program is focused on the development of a new class of steam expanders that offers the potential for significantly lower costs for small-scale power generation technology. The heart of this technology is the scroll expander, a machine that has revolutionized the HVAC refrigerant compressor industry in the last 15 years. The liquid injected cogeneration (LIC) technology is at the core of the efforts described in this report, and remains an excellent option for low cost Micro-CHP systems. ECR has demonstrated in several prototype appliances that the concept for LIC can be made into a practical product. The continuing challenge is to identify economical scroll machine designs that will meet the performance and endurance requirements needed for a long life appliance application. This report describes the numerous advances made in this endeavor by ECR International. Several important advances are described in this report. Section 4 describes a marketing and economics study that integrates the technical performance of the LIC system with real-world climatic data and economic analysis to assess the practical impact that different factors have on the economic application of Micro-CHP in residential applications. Advances in the development of a working scroll steam expander are discussed in Section 5. A rigorous analytical assessment of the performance of scroll expanders, including the difficult to characterize impact of pocket to pocket flank leakage, is presented in Section 5.1. This is followed with an FEA study of the thermal and pressure induced deflections that would result from the normal operation of an advanced scroll expander. Section 6 describes the different scroll expanders and test fixtures developed during this effort. Another key technical

  15. Development of a UAV-based Global Ozone Lidar Demonstrator (GOLD)

    NASA Astrophysics Data System (ADS)

    Browell, E. V.; Deyoung, R. J.; Hair, J. W.; Ismail, S.; McGee, T.; Hardesty, R. M.; Brewer, W. A.; McDermid, I. S.

    2006-12-01

    Global ozone measurements are needed across the troposphere with high vertical resolution to enable comprehensive studies of continental and intercontinental atmospheric chemistry and dynamics, which are affected by diverse natural and human-induced processes. The development of a unattended aerial vehicle (UAV) based Global Ozone Lidar Demonstrator (GOLD) is an important step in enabling a space-based ozone and aerosol lidar and for conducting unique UAV-based large-scale atmospheric investigations. The GOLD system will incorporate the most advanced technology developed under the NASA Laser Risk Reduction Program (LRRP) and the Small Business Innovative Research (SBIR) program to produce a compact, autonomously operating ozone and aerosol Differential Absorption Lidar (DIAL) system for a UAV platform. This system will leverage advanced Nd:YAG and optical parametric oscillator (OPO) laser technologies being developed by ITT Industries under the LRRP and the autonomously operating ozone DIAL system being developed by Science and Engineering Services Inc. (SESI) under an SBIR Phase-3 contract. Laser components from ITT will be integrated into the SESI DIAL system, and the resulting GOLD system will be flight tested on a NASA UAV. The development of the GOLD system was initiated as part of the NASA Instrument Incubator Program in December 2005, and great progress has been made towards completing major GOLD subsystems. ITT has begun construction of the high-power Nd:YAG pump laser and the ultraviolet OPO for generating the ozone DIAL wavelengths of 290 and 300 nm and the aerosol visible wavelength at 532 nm. SESI is completing the Phase-3 SBIR contract for the delivery and demonstration of the ozone DIAL receiver and data system, and NOAA is completing detector evaluations for use in the GOLD system. Welch Mechanical is examining system designs for integrating GOLD into the external pod that will be hung under the new IKANA (Predator-B) UAV that NASA Dryden is

  16. Industrial advanced turbine systems: Development and demonstration. Quarterly report, January 1--March 31, 1998

    SciTech Connect

    1998-08-01

    The US Department of Energy (DOE) has initiated a program for advanced turbine systems (ATS) that will serve industrial power generation markets. The objective of the cooperative agreements granted under the program is to join the DOE with industry in research and development that will lead to commercial offerings in the private sector. The ATS will provide ultra-high efficiency, environmental superiority, and cost competitiveness. The ATS will foster (1) early market penetration that enhances the global competitiveness of US industry, (2) public health benefits resulting from reduced exhaust gas emissions of target pollutants, (3) reduced cost of power used in the energy-intensive industrial marketplace, and (4) the retention and expansion of the skilled US technology base required for the design, development and maintenance of state-of-the-art advanced turbine products. The Industrial ATS Development and Demonstration program is a multi-phased effort. Solar Turbines Incorporated (Solar) has participated in Phases 1 and 2 of the program. On September 14, 1995 Solar was awarded a Cooperative Agreement for Phases 3 and 4 of the program. Phase 3 of the work is separated into two subphases: Phase 3A entails Component Design and Development; Phase 3B will involve Integrated Subsystem Testing. Phase 4 will cover Host Site Testing. As of the end of the reporting period work on the program is 29.1% complete (24.7% last quarter). Work on the Mercury 50 development and ATS technology development portions of the program (WBS 10000 et seq) is 48.9% complete (41.6% last quarter). Estimates of percent complete are based upon milestones completed. In order to maintain objectivity in assessing schedule progress, Solar uses a 0/100 percent complete assumption for milestones rather than subjectively estimating progress toward completion of milestones. Cost and schedule variance information is provided in Section 4.0 Program Management.

  17. Molecular evidence for the coordination of nitrogen and carbon metabolisms, revealed by a study on the transcriptional regulation of the agl3EFG operon that encodes a putative carbohydrate transporter in Streptomyces coelicolor.

    PubMed

    Cen, Xu-Feng; Wang, Jing-Zhi; Zhao, Guo-Ping; Wang, Ying; Wang, Jin

    2016-03-18

    In the agl3EFGXYZ operon (SCO7167-SCO7162, abbreviated as agl3 operon) of Streptomyces coelicolor M145, agl3EFG genes encode a putative ABC-type carbohydrate transporter. The transcription of this operon has been proved to be repressed by Agl3R (SCO7168), a neighboring GntR-family regulator, and this repression can be released by growth on poor carbon sources. Here in this study, we prove that the transcription of agl3 operon is also directly repressed by GlnR, a central regulator governing the nitrogen metabolism in S. coelicolor. The electrophoretic mobility shift assay (EMSA) employing the agl3 promoter and mixtures of purified recombinant GlnR and Agl3R indicates that GlnR and Agl3R bind to different DNA sequences within the promoter region of agl3 operon, which is further confirmed by the DNase I footprinting assay. As Agl3R and GlnR have been demonstrated to sense the extracellular carbon and nitrogen supplies, respectively, it is hypothesized that the transcription of agl3 operon is stringently governed by the availabilities of extracellular carbon and nitrogen sources. Consistent with the hypothesis, the agl3 operon is further found to be derepressed only under the condition of poor carbon and rich nitrogen supplies, when both regulators are inactivated. It is believed that activation of the expression of agl3 operon may facilitate the absorption of extracellular carbohydrates to balance the ratio of intracellular carbon to nitrogen.

  18. Development/Demonstration of an Advanced Oxy-Fuel Front-End System

    SciTech Connect

    Mighton, Steven, J.

    2007-08-06

    Owens Corning and other glass manufacturers have used oxy-fuel combustion technology successfully in furnaces to reduce emissions, increase throughput, reduce fuel consumption and, depending on the costs of oxygen and fuel, reduce energy costs. The front end of a fiberglass furnace is the refractory channel system that delivers glass from the melter to the forming process. After the melter, it is the second largest user of energy in a fiberglass plant. A consortium of glass companies and suppliers, led by Owens Corning, was formed to develop and demonstrate oxy/fuel combustion technology for the front end of a fiberglass melter, to demonstrate the viability of this energy saving technology to the U.S. glass industry, as a D.O.E. sponsored project. The project goals were to reduce natural gas consumption and CO2 green house gas emissions by 65 to 70% and create net cost savings after the purchase of oxygen to achieve a project payback of less than 2 years. Project results in Jackson, TN included achieving a 56% reduction in gas consumption and CO2 emissions. A subsequent installation in Guelph ON, not impacted by unrelated operational changes in Jackson, achieved a 64% reduction. Using the more accurate 64% reduction in the payback calculation yielded a 2.2 year payback in Jackson. The installation of the demonstration combustion system saves 77,000 DT/yr of natural gas or 77 trillion Btu/yr and eliminates 4500 tons/yr of CO2 emissions. This combustion system is one of several energy and green house gas reduction technologies being adopted by Owens Corning to achieve aggressive goals relating to the company’s global facility environmental footprint.

  19. Laboratory development and field demonstration of self-sealing/self-healing landfill liner.

    PubMed

    Shi, Caijun; Booth, Rob

    2005-01-01

    The self-sealing/self-healing (SS/SH) barrier concept is based on the principle that two or more parent materials placed in vertical or horizontal layers will react at their interfaces to form insoluble reaction products. These products constitute a seamless impermeable seal, which is resistant to the transmission of leachate and contaminants. A SS/SH liner formulation was developed in the laboratory and demonstrated at the Sudokwon landfill site in South Korea. Laboratory testing results indicated that a seal with a hydraulic conductivity less than 10(-9) m/s formed after two to four weeks of curing at room temperature, and the seal healed itself after it was fractured. The use of the soil from the Sudokwon landfill site instead of sand as the matrix of the parent materials in the SS/SH liner retarded the sealing and healing of the seal, but did not show an obvious effect on the overall sealing and healing capacity of the seal at early stages. The construction and installation of the field demonstration SS/SH liner were carried out in the same way as for a soil cement liner. The quality of the liner was ensured by the enforcement of quality analysis/quality control procedures during installation. A single sealed ring infiltration test was performed on the field demonstration liner 36 days after the installation was completed. The measurement of water infiltration rate indicated that the liner healed after it was fractured. However, the long-term sealing and healing capacity needs to be further investigated.

  20. Industrial Advanced Turbine Systems: Development and Demonstration. Annual report, September 14, 1995--September 30, 1996

    SciTech Connect

    1998-12-31

    The U.S. Department of Energy (DOE) has initiated a program for advanced turbine systems (ATS) that will serve industrial power generation markets. The objective of the cooperative agreements granted under the program is to join the DOE with industry in research and development that will lead to commercial offerings in the private sector. The ATS will provide ultra-high efficiency, environmental superiority, and cost competitiveness. The ATS will foster (1) early market penetration that enhances the global competitiveness of U.S. industry, (2) public health benefits resulting from reduced exhaust gas emissions of target pollutants, (3) reduced cost of power used in the energy-intensive industrial marketplace and (4) the retention and expansion of the skilled U.S. technology base required for the design, development and maintenance of state-of-the-art advanced turbine products. The Industrial ATS Development and Demonstration program is a multi-phased effort. Solar Turbines Incorporated (Solar) has participated in Phases 1 and 2 of the program. On September 14, 1995 Solar was awarded a Cooperative Agreement for Phases 3 and 4 of the program (DE-FC21-95MC31173) by the DOE`s Office of Energy Efficiency and Renewable Energy (EE). Technical administration of the Cooperative Agreement will be provided from EE`s Chicago Operations Office. Contract administration of the Cooperative Agreement will be provided from DOE`s Office of Fossil Energy, Morgantown Energy Technology Center (METC).

  1. Development and pilot demonstration program of a waste minimization plan at Argonne National Laboratory

    SciTech Connect

    Peters, R.W.; Wentz, C.A.; Thuot, J.R.

    1991-01-01

    In response to US Department of Energy directives, Argonne National Laboratory (ANL) has developed a waste minimization plan aimed at reducing the amount of wastes at this national research and development laboratory. Activities at ANL are primarily research- oriented and as such affect the amount and type of source reduction that can be achieved at this facility. The objective of ANL's waste minimization program is to cost-effectively reduce all types of wastes, including hazardous, mixed, radioactive, and nonhazardous wastes. The ANL Waste Minimization Plan uses a waste minimization audit as a systematic procedure to determine opportunities to reduce or eliminate waste. To facilitate these audits, a computerized bar-coding procedure is being implemented at ANL to track hazardous wastes from where they are generated to their ultimate disposal. This paper describes the development of the ANL Waste Minimization Plan and a pilot demonstration of the how the ANL Plan audited the hazardous waste generated within a selected divisions of ANL. It includes quantitative data on the generation and disposal of hazardous waste at ANL and describes potential ways to minimize hazardous wastes. 2 refs., 5 figs., 8 tabs.

  2. Experimental Development and Demonstration of Ultrasonic Measurement Diagnostics for Sodium Fast Reactor Thermal-hydraulics

    SciTech Connect

    Tokuhiro, Akira; Jones, Byron

    2013-09-13

    This research project will address some of the principal technology issues related to sodium-cooled fast reactors (SFR), primarily the development and demonstration of ultrasonic measurement diagnostics linked to effective thermal convective sensing under normatl and off-normal conditions. Sodium is well-suited as a heat transfer medium for the SFR. However, because it is chemically reactive and optically opaque, it presents engineering accessibility constraints relative to operations and maintenance (O&M) and in-service inspection (ISI) technologies that are currently used for light water reactors. Thus, there are limited sensing options for conducting thermohydraulic measurements under normal conditions and off-normal events (maintenance, unanticipated events). Acoustic methods, primarily ultrasonics, are a key measurement technology with applications in non-destructive testing, component imaging, thermometry, and velocimetry. THis project would have yielded a better quantitative and qualitative understanding of the thermohydraulic condition of solium under varied flow conditions. THe scope of work will evaluate and demonstrate ultrasonic technologies and define instrumentation options for the SFR.

  3. 3-d finite element model development for biomechanics: a software demonstration

    SciTech Connect

    Hollerbach, K.; Hollister, A.M.; Ashby, E.

    1997-03-01

    Finite element analysis is becoming an increasingly important part of biomechanics and orthopedic research, as computational resources become more powerful, and data handling algorithms become more sophisticated. Until recently, tools with sufficient power did not exist or were not accessible to adequately model complicated, three-dimensional, nonlinear biomechanical systems. In the past, finite element analyses in biomechanics have often been limited to two-dimensional approaches, linear analyses, or simulations of single tissue types. Today, we have the resources to model fully three-dimensional, nonlinear, multi-tissue, and even multi-joint systems. The authors will present the process of developing these kinds of finite element models, using human hand and knee examples, and will demonstrate their software tools.

  4. Research, development, and demonstration of nickel-iron batteries for electric vehicle propulsion. Annual report, 1980

    SciTech Connect

    Not Available

    1981-03-01

    The objective of the Eagle-Picher nickel-iron battery program is to develop a nickel-iron battery for use in the propulsion of electric and electric-hybrid vehicles. To date, the program has concentrated on the characterization, fabrication and testing of the required electrodes, the fabrication and testing of full-scale cells, and finally, the fabrication and testing of full-scale (270 AH) six (6) volt modules. Electrodes of the final configuration have now exceeded 1880 cycles and are showing minimal capacity decline. Full-scale cells have presently exceeded 600 cycles and are tracking the individual electrode tests almost identically. Six volt module tests have exceeded 500 cycles, with a specific energy of 48 Wh/kg. Results to date indicate the nickel-iron battery is beginning to demonstrate the performance required for electric vehicle propulsion.

  5. Hybrid Neural-Network: Genetic Algorithm Technique for Aircraft Engine Performance Diagnostics Developed and Demonstrated

    NASA Technical Reports Server (NTRS)

    Kobayashi, Takahisa; Simon, Donald L.

    2002-01-01

    As part of the NASA Aviation Safety Program, a unique model-based diagnostics method that employs neural networks and genetic algorithms for aircraft engine performance diagnostics has been developed and demonstrated at the NASA Glenn Research Center against a nonlinear gas turbine engine model. Neural networks are applied to estimate the internal health condition of the engine, and genetic algorithms are used for sensor fault detection, isolation, and quantification. This hybrid architecture combines the excellent nonlinear estimation capabilities of neural networks with the capability to rank the likelihood of various faults given a specific sensor suite signature. The method requires a significantly smaller data training set than a neural network approach alone does, and it performs the combined engine health monitoring objectives of performance diagnostics and sensor fault detection and isolation in the presence of nominal and degraded engine health conditions.

  6. Affordable Development and Demonstration of a Small NTR Engine and Stage: How Small is Big Enough?

    NASA Technical Reports Server (NTRS)

    Borowski, Stanley K.; Sefcik, Robert J.; Fittje, James E.; McCurdy, David R.; Qualls, Arthur L.; Schnitzler, Bruce G.; Werner, James E.; Weitzberg (Abraham); Joyner, Claude R.

    2015-01-01

    The Nuclear Thermal Rocket (NTR) derives its energy from fission of uranium-235 atoms contained within fuel elements that comprise the engine's reactor core. It generates high thrust and has a specific impulse potential of approximately 900 seconds - a 100% increase over today's best chemical rockets. The Nuclear Thermal Propulsion (NTP) project, funded by NASA's AES program, includes five key task activities: (1) Recapture, demonstration, and validation of heritage graphite composite (GC) fuel (selected as the "Lead Fuel" option); (2) Engine Conceptual Design; (3) Operating Requirements Definition; (4) Identification of Affordable Options for Ground Testing; and (5) Formulation of an Affordable Development Strategy. During FY'14, a preliminary DDT&E plan and schedule for NTP development was outlined by GRC, DOE and industry that involved significant system-level demonstration projects that included GTD tests at the NNSS, followed by a FTD mission. To reduce cost for the GTD tests and FTD mission, small NTR engines, in either the 7.5 or 16.5 klbf thrust class, were considered. Both engine options used GC fuel and a "common" fuel element (FE) design. The small approximately 7.5 klbf "criticality-limited" engine produces approximately 157 megawatts of thermal power (MWt) and its core is configured with parallel rows of hexagonal-shaped FEs and tie tubes (TTs) with a FE to TT ratio of approximately 1:1. The larger approximately 16.5 klbf Small Nuclear Rocket Engine (SNRE), developed by LANL at the end of the Rover program, produces approximately 367 MWt and has a FE to TT ratio of approximately 2:1. Although both engines use a common 35 inch (approximately 89 cm) long FE, the SNRE's larger diameter core contains approximately 300 more FEs needed to produce an additional 210 MWt of power. To reduce the cost of the FTD mission, a simple "1-burn" lunar flyby mission was considered to reduce the LH2 propellant loading, the stage size and complexity. Use of existing and

  7. A comparison of key aspects of gene regulation in Streptomyces coelicolor and Escherichia coli using nucleotide-resolution transcription maps produced in parallel by global and differential RNA sequencing

    PubMed Central

    Romero, David A; Hasan, Ayad H; Lin, Yu-fei; Kime, Louise; Ruiz-Larrabeiti, Olatz; Urem, Mia; Bucca, Giselda; Mamanova, Lira; Laing, Emma E; van Wezel, Gilles P; Smith, Colin P; Kaberdin, Vladimir R; McDowall, Kenneth J

    2014-01-01

    Streptomyces coelicolor is a model for studying bacteria renowned as the foremost source of natural products used clinically. Post-genomic studies have revealed complex patterns of gene expression and links to growth, morphological development and individual genes. However, the underlying regulation remains largely obscure, but undoubtedly involves steps after transcription initiation. Here we identify sites involved in RNA processing and degradation as well as transcription within a nucleotide-resolution map of the transcriptional landscape. This was achieved by combining RNA-sequencing approaches suited to the analysis of GC-rich organisms. Escherichia coli was analysed in parallel to validate the methodology and allow comparison. Previously, sites of RNA processing and degradation had not been mapped on a transcriptome-wide scale for E. coli. Through examples, we show the value of our approach and data sets. This includes the identification of new layers of transcriptional complexity associated with several key regulators of secondary metabolism and morphological development in S. coelicolor and the identification of host-encoded leaderless mRNA and rRNA processing associated with the generation of specialized ribosomes in E. coli. New regulatory small RNAs were identified for both organisms. Overall the results illustrate the diversity in mechanisms used by different bacterial groups to facilitate and regulate gene expression. PMID:25266672

  8. Industrial advanced turbine systems: Development and demonstration. Annual report, October 1, 1996--September 30, 1997

    SciTech Connect

    1997-12-31

    The US DOE has initiated a program for advanced turbine systems (ATS) that will serve industrial power generation markets. The ATS will provide ultra-high efficiency, environmental superiority, and cost competitiveness. The ATS will foster (1) early market penetration that enhances the global competitiveness of US industry, (2) public health benefits resulting from reduced exhaust gas emissions of target pollutants, (3) reduced cost of power used in the energy-intensive industrial marketplace and (4) the retention and expansion of the skilled US technology base required for the design, development and maintenance of state-of-the-art advanced turbine products. The Industrial ATS Development and Demonstration program is a multi-phased effort. Solar Turbines Incorporated (Solar) has participated in Phases 1 and 2 of the program. On September 14, 1995 Solar was awarded a Cooperative Agreement for Phases 3 and 4 of the program. Phase 3 of the work is separated into two subphases: Phase 3A entails Component Design and Development Phase 3B will involve Integrated Subsystem Testing. Phase 4 will cover Host Site Testing. Forecasts call for completion of the program within budget as originally estimated. Scheduled completion is forecasted to be approximately 3 years late to original plan. This delay has been intentionally planned in order to better match program tasks to the anticipated availability of DOE funds. To ensure the timely realization of DOE/Solar program goals, the development schedule for the smaller system (Mercury 50) and enabling technologies has been maintained, and commissioning of the field test unit is scheduled for May of 2000. As of the end of the reporting period work on the program is 22.80% complete based upon milestones completed. This measurement is considered quite conservative as numerous drawings on the Mercury 50 are near release. Variance information is provided in Section 4.0-Program Management.

  9. A1 demonstrates restricted tissue distribution during embryonic development and functions to protect against cell death.

    PubMed Central

    Carrió, R.; López-Hoyos, M.; Jimeno, J.; Benedict, M. A.; Merino, R.; Benito, A.; Fernández-Luna, J. L.; Núñez, G.; García-Porrero, J. A.; Merino, J.

    1996-01-01

    Members of the bcl-2 gene family are essential regulators of cell survival in a wide range of biological processes. A1, a member of the family, is known to be expressed in certain adult tissues. However, the precise tissue distribution and function of A1 remains poorly understood. We show here that A1 is expressed in multiple tissues during murine embryonic development. In the embryo, A1 was detected first at embryonic day 11.5 in liver, brain, and limbs. At day 13.5 of gestation, A1 expression was observed in the central nervous system, liver, perichondrium, and digital zones of developing limbs in a pattern different from that of bcl-X. In the central nervous system of 15.5-day embryos, A1 was expressed at high levels in the ventricular zone and cortical plate of brain cortex. Significantly, the interdigital zones of limbs and the intermediate region of the developing brain cortex, two sites associated with extensive cell death, were devoid of A1 and bcl-X. The expression of A1 was retained in many adult tissues. To assess the ability of A1 to modulate cell death, stable transfectants expressing different amounts of A1 protein were generated in K562 cells. Expression of A1 was associated with retardation of apoptotic cell death induced by actinomycin D and cycloheximide as well as by okadaic acid. Confocal microscopy showed that the A1 protein was localized to the cytoplasm in a pattern similar to that of Bcl-2. These results demonstrate that the expression of A1 is wider than previously reported in adult tissues. Furthermore, its distribution in multiple tissues of the embryo suggests that A1 plays a role in the regulation of physiological cell death during embryonic development. Images Figure 1 Figure 2 Figure 3 Figure 5 PMID:8952545

  10. Study and Development of a Sub-Orbital Re-Entry Demonstrator

    NASA Astrophysics Data System (ADS)

    Savino, R.

    The Italian and European Space Agencies are supporting a research programme, developed in Campania region by a cluster of industries, research institutes and universities, on a low-cost re-entry capsule, able to return payloads from the ISS to Earth and/or to perform short-duration scientific missions in Low Earth Orbit (LEO). The ballistic capsule is characterized by a deployable, disposable "umbrella-like" heat shield that allows relatively small dimensions at launch and a sufficient exposed surface area in re-entry conditions, reducing the ballistic coefficient and leading to acceptable heat fluxes, mechanical loads and final descent velocity. ESA is supporting a preliminary study to develop a flight demonstrator of the capsule to be embarked as a secondary payload onboard a sub-orbital sounding rocket. The deployable thermal protection system concept may be applied to future science and robotic exploration mission requiring planetary entry and, possibly also to missions in the framework of Human Space flight, requiring planetary entry or re-entry. The technology offers also an interesting potential for aerobraking, aerocapture and for de-orbiting. This paper summarizes the results of these activities, which are being more and more refined as the work proceeds, including the definition and analysis of the mission scenario, the aerodynamic, aerothermodynamic, mechanical and structural analyses and the technical definition of avionics, instrumentation and main subsystems.

  11. Interspecies modulation of bacterial development through iron competition and siderophore piracy.

    PubMed

    Traxler, Matthew F; Seyedsayamdost, Mohammad R; Clardy, Jon; Kolter, Roberto

    2012-11-01

    While soil-dwelling actinomycetes are renowned for secreting natural products, little is known about the roles of these molecules in mediating actinomycete interactions. In a previous co-culture screen, we found that one actinomycete, Amycolatopsis sp. AA4, inhibited aerial hyphae formation in adjacent colonies of Streptomyces coelicolor. A siderophore, amychelin, mediated this developmental arrest. Here we present genetic evidence that confirms the role of the amc locus in the production of amychelin and in the inhibition of S. coelicolor development. We further characterize the Amycolatopsis sp. AA4 - S. coelicolor interaction by examining expression of developmental and iron acquisition genes over time in co-culture. Manipulation of iron availability and/or growth near Amycolatopsis sp. AA4 led to alterations in expression of the critical developmental gene bldN, and other key downstream genes in the S. coelicolor transcriptional cascade. In Amycolatopsis sp. AA4, siderophore genes were downregulated when grown near S. coelicolor, leading us to find that deferrioxamine E, produced by S. coelicolor, could be readily utilized by Amycolatopsis sp. AA4. Collectively these results suggest that competition for iron via siderophore piracy and species-specific siderophores can alter patterns of gene expression and morphological differentiation during actinomycete interactions.

  12. Industrial advanced turbine systems: Development and demonstration. Quarterly report, July 1--September 30, 1997

    SciTech Connect

    1997-12-31

    The US DOE has initiated a program for advanced turbine systems (ATS) that will serve industrial power generation markets. The ATS will foster (1) early market penetration that enhances the global competitiveness of US industry, (2) public health benefits resulting from reduced exhaust gas emissions of target pollutants, (3) reduced cost of power used in the energy-intensive industrial marketplace and (4) the retention and expansion of the skilled US technology base required for the design, development and maintenance of state-of-the-art advanced turbine products. The Industrial ATS Development and Demonstration program is a multi-phased effort. Solar Turbines Incorporated (Solar) has participated in Phases 1 and 2 of the program. On September 14, 1995 Solar was awarded a Cooperative Agreement for Phases 3 and 4 of the program. Phase 3 of the work is separated into two subphases: Phase 3A entails Component Design and Development; Phase 3B will involve Integrated Subsystem Testing. Phase 4 will cover Host Site Testing. Forecasts call for completion of the program within budget as originally estimated. Scheduled completion is forecasted to be approximately 3 years late to original plan. Significant efforts were spent this quarter to reforecast and control expenditures due to Solar`s and DOE`s current funding and resource constraints. Selective reductions and delays in program activities were identified and implemented. Although these actions will increase technical risk and the attainment of stretch goals, it is not anticipated that the schedule for initial test units or the attainment of basic program performance requirements will be impacted. As of the end of the reporting period work on the program is 22.80% complete based upon milestones completed. This measurement is considered quite conservative as numerous drawings on the Mercury 50 are near release. Variance information is provided in Section 4.0-Program Management.

  13. ROSE: development and demonstration of a "Mobile Response Observatory" prototype for subsea environmental monitoring.

    NASA Astrophysics Data System (ADS)

    Marvaldi, J.; Legrand, J.; Masset, J. F.

    2009-04-01

    ROSE project was aimed at developing an underwater monitoring system deployable in situations of crisis such as wreck pollution or risky natural sea bottom events. The system is based on subsea stations integrated in an acoustic network and communicating with shore through a radio link.. The project was supported by the French Research Ministry. Based on detailed functional and system specifications, a prototype system composed of two measuring stations and a relay buoy was built and tested during 2.5 months in coastal area. Subsea stations developed and built by Ifremer are anchored some meters above sea bottom and make it possible to position sensors at a convenient height e.g. in accordance with pollutant emissions. Stations are structured in several areas dedicated to the various equipment pieces : electronics and energy containers, sensor area, floatation volumes, free area aimed at specific uses. The buoy is derived from designs already experienced by Ifremer. The bi-directional communication system was developed and implemented by Sercel UAD. The acoustic network is of MATS200 Net. Radio communication between the operating station onshore and the relay buoy consists of a radio link of cellular phone, VHF radio or satellite type depending on deployment site . Data registered by the stations are periodically transmitted onshore and stored in a data server. For the prototype demonstration, the radio link was of GPRS phone type. NKE has developed the messengers, which are positively buoyant beacons enabling the retrieval of data in addition to or in absence of the acoustic-radio communication system. Up to 8 messengers can be stored on a station. Messengers successively receive summaries of registered data and can be released either at predefined time steps or on a command. Once at sea surface they transmit the data via Argos system. Station design enables easy changing of sensor set. In case of pollution event the set is composed of pollutant and environmental

  14. Development of the Decontamination Approach for the West Valley Demonstration Project Decontamination Project Plan

    SciTech Connect

    Milner, T. N.; Watters, W. T.

    2002-02-25

    This paper details the development of a decontamination approach for the West Valley Demonstration Project (WVDP), Decontamination Project Plan (Plan). The WVDP is operated by West Valley Nuclear Services Company (WVNSCO), a subsidiary of Westinghouse Government and Environmental Services, and its parent companies Washington Group International and British Nuclear Fuels Limited (BNFL). The WVDP is a waste management effort being conducted by the United States Department of Energy (DOE) at the site of the only commercial nuclear fuel reprocessing facility to have operated in the United States. This facility is part of the Western New York Nuclear Service Center (WNYNSC), which is owned by the New York State Energy Research and Development Authority (NYSERDA). As authorized by Congress in 1980 through the West Valley Demonstration Project Act (WVDP Act, Public Law 96-368), the DOE's primary mission at the WVDP is to solidify high-level liquid nuclear waste safely; transport the high-level waste (HLW) to a federal repository; and decontaminate and decommission the facilities and hardware used to solidify the HLW and conduct the WVDP. This includes a provision for the disposal of low-level waste (LLW) and transuranic waste (TRU) produced during processing of the HLW. Continuation of the effort to reduce the hazard and risk associated with historic operations to the extent needed to ensure the health and safety of the public and the environment will see a change in focus from stabilization of liquid HLW to stabilization of former plutonium and uranium extraction (PUREX) reprocessing plant facilities. This will be achieved through the activities of in-cell component removal and packaging, and preparation for long-term disposal of the long- lived radionuclides. These radionuclides are associated with the former PUREX facility operations, including, and upstream from, facilities utilized in the primary separation and first plutonium/uranium split cycles. The closure

  15. Proteasome involvement in a complex cascade mediating SigT degradation during differentiation of Streptomyces coelicolor.

    PubMed

    Mao, Xu-Ming; Ren, Ni-Ni; Sun, Ning; Wang, Feng; Zhou, Ri-Cheng; Tang, Yi; Li, Yong-Quan

    2014-02-14

    In Streptomyces coelicolor, the ECF sigma factor SigT negatively regulates cell differentiation, and is degraded by ClpP protease in a dual positive feedback manner. Here we further report that the proteasome is required for degradation of SigT, but not for degradation of its anti-sigma factor RstA, and RstA can protect SigT from degradation independent of the proteasome. Meanwhile, deletion of the proteasome showed reduced production of secondary metabolites, and the fermentation medium from wild type could promote SigT degradation. Furthermore, overexpression of redD or actII-orf4 in the proteasome-deficiency mutant resulted in SigT degradation and over-production of both undecylprodigiosin and actinorhodin. Therefore the proteasome is required for SigT degradation by affecting the production of secondary metabolites during cell differentiation.

  16. Genetic analysis of absB, a Streptomyces coelicolor locus involved in global antibiotic regulation.

    PubMed

    Adamidis, T; Champness, W

    1992-07-01

    The filamentous soil bacterium Streptomyces coelicolor is known to produce four antibiotics which are genetically and structurally distinct. An extensive search for antibiotic regulatory mutants led to the discovery of absB mutants, which are antibiotic deficient but sporulation proficient. Genetic analysis of the absB mutants has resulted in definition of the absB locus at 5 o'clock on the genetic map. Multiple cloned copies of the actII-ORF4 gene, an activator of synthesis of the antibiotic actinorhodin, restore actinorhodin biosynthetic capability to the absB mutants. These results are interpreted to mean that the failure of absB mutants to produce antibiotics results from decreased expression of the antibiotic genes. The absB gene is proposed to be involved in global regulation of antibiotic synthesis.

  17. Real Options Valuation of U.S. Federal Renewable Energy Research,Development, Demonstration, and Deployment

    SciTech Connect

    Siddiqui, Afzal S.; Marnay, Chris; Wiser, Ryan H.

    2005-03-01

    Benefits analysis of US Federal government funded research, development, demonstration, and deployment (RD3) programs for renewable energy (RE) technology improvement typically employs a deterministic forecast of the cost and performance of renewable and nonrenewable fuels. The benefits estimate for a program derives from the difference between two forecasts, with and without the RD3 in place. The deficiencies of the current approach are threefold: (1) it does not consider uncertainty in the cost of non-renewable energy (NRE), and the option or insurance value of deploying RE if and when NRE costs rise; (2) it does not consider the ability of the RD3 manager to adjust the RD3 effort to suit the evolving state of the world, and the option value of this flexibility; and (3) it does not consider the underlying technical risk associated with RD3, and the impact of that risk on the programs optimal level of RD3 effort. In this paper, a rudimentary approach to determining the option value of publicly funded RE RD3 is developed. The approach seeks to tackle the first deficiency noted above by providing an estimate of the options benefit of an RE RD3 program in a future with uncertain NRE costs.While limited by severe assumptions, a computable lattice of options values reveals the economic intuition underlying the decision-making process. An illustrative example indicates how options expose both the insurance and timing values inherent in a simplified RE RD3 program that coarsely approximates the aggregation of current Federal RE RD3.This paper also discusses the severe limitations of this initial approach, and identifies needed model improvements before the approach can adequately respond to the RE RD3 analysis challenge.

  18. Development of analytical cell support for vitrification at the West Valley Demonstration Project. Topical report

    SciTech Connect

    Barber, F.H.; Borek, T.T.; Christopher, J.Z.

    1997-12-01

    Analytical and Process Chemistry (A&PC) support is essential to the high-level waste vitrification campaign at the West Valley Demonstration Project (WVDP). A&PC characterizes the waste, providing information necessary to formulate the recipe for the target radioactive glass product. High-level waste (HLW) samples are prepared and analyzed in the analytical cells (ACs) and Sample Storage Cell (SSC) on the third floor of the main plant. The high levels of radioactivity in the samples require handling them in the shielded cells with remote manipulators. The analytical hot cells and third floor laboratories were refurbished to ensure optimal uninterrupted operation during the vitrification campaign. New and modified instrumentation, tools, sample preparation and analysis techniques, and equipment and training were required for A&PC to support vitrification. Analytical Cell Mockup Units (ACMUs) were designed to facilitate method development, scientist and technician training, and planning for analytical process flow. The ACMUs were fabricated and installed to simulate the analytical cell environment and dimensions. New techniques, equipment, and tools could be evaluated m in the ACMUs without the consequences of generating or handling radioactive waste. Tools were fabricated, handling and disposal of wastes was addressed, and spatial arrangements for equipment were refined. As a result of the work at the ACMUs the remote preparation and analysis methods and the equipment and tools were ready for installation into the ACs and SSC m in July 1995. Before use m in the hot cells, all remote methods had been validated and four to eight technicians were trained on each. Fine tuning of the procedures has been ongoing at the ACs based on input from A&PC technicians. Working at the ACs presents greater challenges than had development at the ACMUs. The ACMU work and further refinements m in the ACs have resulted m in a reduction m in analysis turnaround time (TAT).

  19. Development and Demonstration of Insulated Pressure Vessels for Vehicular Hydrogen Storage

    SciTech Connect

    Berry, G D; Aceves, S M

    2004-02-26

    This paper describes the development of an alternative technology for vehicular storage of hydrogen. Insulated pressure vessels are cryogenic-capable pressure vessels that can accept cryogenic liquid fuel, cryogenic compressed gas or compressed gas at ambient temperature. Insulated pressure vessels offer advantages over alternative hydrogen storage technologies. Insulated pressure vessels are more compact and less expensive than compressed hydrogen vessels. They have lower evaporative losses and lower energy requirement for fuel liquefaction than liquid hydrogen tanks, and they are lighter than hydrides. The work described in this paper is directed at verifying that insulated pressure vessels can be used safely for vehicular hydrogen storage. The paper describes multiple tests and analyses that have been conducted to evaluate the safety of insulated pressure vessels. Insulated pressure vessels have been subjected to multiple DOT, ISO and SAE certification tests, and the vessels have always been successful in meeting the passing criteria for the different tests. A draft procedure for insulated pressure vessel certification has been generated to assist in a future commercialization of this technology. Ongoing work includes the demonstration of this technology in a vehicle.

  20. Demonstration Model Development of the Force-Balanced Coil for SMES

    NASA Astrophysics Data System (ADS)

    Nomura, Shinichi; Kasuya, Koji; Tanaka, Norihiro; Tsuboi, Kenji; Tsutsui, Hiroaki; Shimada, Ryuichi; Ninomiya, Akira; Ishigohka, Takashi

    In large-scale SMES, the superconducting coils require special considerations for induced electromagnetic forces to limit allowable tensile stress. Force-balanced coil (FBC) is a helically wound hybrid coil of toroidal field coils and a solenoid. The FBC can significantly reduce the required mass of the structure for induced electromagnetic forces. In order to demonstrate the feasibility of the FBC concept for SMES, the authors have developed a superconducting model coil. The outer diameter of the model FBC is 0.53m. The hand-made winding, using NbTi/Cu composite strands with a diameter of 1.17mm, was finished with 10584 poloidal turns after four months. The helical windings of the model FBC were neither impregnated with epoxy resin nor reinforced with stainless steel wires. Three test runs were conducted with liquid helium cooling at intervals of several months. The number of quench tests was 81 in total. The first quench current was 293A, which was 53% of the critical coil current. The training phenomena could be observed even after the coil was warmed up to room temperature. After successive quenches the quench current was improved to 476A, corresponding to 86% of the critical coil current, and it was successfully excited up to 6.1T.

  1. VISDTA: A video imaging system for detection, tracking, and assessment: Prototype development and concept demonstration

    SciTech Connect

    Pritchard, D.A.

    1987-05-01

    It has been demonstrated that thermal imagers are an effective surveillance and assessment tool for security applications because: (1) they work day or night due to their sensitivity to thermal signatures; (2) penetrability through fog, rain, dust, etc., is better than human eyes; (3) short or long range operation is possible with various optics; and (4) they are strictly passive devices providing visible imagery which is readily interpreted by the operator with little training. Unfortunately, most thermal imagers also require the setup of a tripod, connection of batteries, cables, display, etc. When this is accomplished, the operator must manually move the camera back and forth searching for signs of aggressor activity. VISDTA is designed to provide automatic panning, and in a sense, ''watch'' the imagery in place of the operator. The idea behind the development of VISDTA is to provide a small, portable, rugged system to automatically scan areas and detect targets by computer processing of images. It would use a thermal imager and possibly an intensified day/night TV camera, a pan/ tilt mount, and a computer for system control. If mounted on a dedicated vehicle or on a tower, VISDTA will perform video motion detection functions on incoming video imagery, and automatically scan predefined patterns in search of abnormal conditions which may indicate attempted intrusions into the field-of-regard. In that respect, VISDTA is capable of improving the ability of security forces to maintain security of a given area of interest by augmenting present techniques and reducing operator fatigue.

  2. Development of simulated tank wastes for the US Department of Energy's Underground Storage Tank Integrated Demonstration

    SciTech Connect

    Elmore, M.R.; Colton, N.G.; Jones, E.O.

    1992-08-01

    The purpose of the Underground Storage Tank Integrated Demonstration (USTID) is to identify and evaluate technologies that may be used to characterize, retrieve, treat, and dispose of hazardous and radioactive wastes contained in tanks on US Department of Energy sites. Simulated wastes are an essential component of the evaluation process because they provide controlled samples for technology assessment, and minimize costs and risks involved when working with radioactive wastes. Pacific Northwest Laboratory has developed a recipe to simulate Hanford single-shell tank, (SST) waste. The recipe is derived from existing process recipes, and elemental concentrations are based on characterization data from 18 SSTs. In this procedure, salt cake and metal oxide/hydroxide sludge are prepared individually, and mixed together at varying ratios depending on the specific tank, waste to be simulated or the test being conducted. Elemental and physical properties of the stimulant are comparable with analyzed tank samples, and chemical speciation in the simulant is being improved as speciation data for actual wastes become available. The nonradioactive chemical waste simulant described here is useful for testing technologies on a small scale.

  3. GLASS FORMULATION DEVELOPMENT TO SUPPORT MELTER TESTING TO DEMONSTRATE ENHANCED HIGH LEVEL WASTE THROUGHPUT

    SciTech Connect

    Marra, J; David Peeler, D; Tommy Edwards, T; Kevin Fox, K; Amanda Youchak, A; James Gillam, J

    2007-08-17

    The U.S. Department of Energy (DOE) is currently processing high-level waste (HLW) through a Joule-heated melter (JHM) at the Savannah River Site (SRS) and plans to vitrify HLW and Low activity waste (LAW) at the Hanford Site. Over the past few years at the DWPF, work has concentrated on increasing waste throughput. These efforts are continuing with an emphasis on high alumina content feeds. High alumina feeds have presented specific challenges for the JHM technology regarding the ability to increase waste loading yet still maintain product quality and adequate throughput. Alternatively, vitrification technology innovations are also being investigated as a means to increase waste throughput. The Cold Crucible Induction Melter (CCIM) technology affords the opportunity for higher vitrification process temperatures as compared to the current reference JHM technology. Higher process temperatures may allow for higher waste loading and higher melt rate. Glass formulation testing to support melter demonstration testing was recently completed. This testing was specifically aimed at high alumina concentration wastes. Glass composition property models were utilized as a guide for formulation development. Both CCIM and JHM testing will be conducted so glass formulation testing was targeted at both technologies with a goal to significantly increase waste loading without compromising product quality.

  4. Accelerator production of tritium plant design and supporting engineering development and demonstration work

    SciTech Connect

    Lisowski, P.W.

    1997-11-01

    Tritium is an isotope of hydrogen with a half life of 12.3 years. Because it is essential for US thermonuclear weapons to function, tritium must be periodically replenished. Since K reactor at Savannah River Site stopped operating in 1988, tritium has been recycled from dismantled nuclear weapons. This process is possible only as long as many weapons are being retired. Maintaining the stockpile at the level called for in the present Strategic Arms Reduction Treaty (START-I) will require the Department of Energy to have an operational tritium production capability in the 2005--2007 time frame. To make the required amount of tritium using an accelerator based system (APT), neutrons will be produced through high energy proton reactions with tungsten and lead. Those neutrons will be moderated and captured in {sup 3}He to make tritium. The APT plant design will use a 1,700 MeV linear accelerator operated at 100 mA. In preparation for engineering design, starting in October 1997 and subsequent construction, a program of engineering development and demonstration is underway. That work includes assembly and testing of the first 20 MeV of the low energy plant linac at 100 mA, high-energy linac accelerating structure prototyping, radiofrequency power system improvements, neutronic efficiency measurements, and materials qualifications.

  5. Development of a Ground Operations Demonstration Unit for Liquid Hydrogen at Kennedy Space Center

    NASA Astrophysics Data System (ADS)

    Notardonato, W. U.

    NASA operations for handling cryogens in ground support equipment have not changed substantially in 50 years, despite major technology advances in the field of cryogenics. NASA loses approximately 50% of the hydrogen purchased because of a continuous heat leak into ground and flight vessels, transient chill down of warm cryogenic equipment, liquid bleeds, and vent losses. NASA Kennedy Space Center (KSC) needs to develop energy-efficient cryogenic ground systems to minimize propellant losses, simplify operations, and reduce cost associated with hydrogen usage. The GODU LH2 project will design, assemble, and test a prototype storage and distribution system for liquid hydrogen that represents an advanced end-to-end cryogenic propellant system for a ground launch complex. The project has multiple objectives and will culminate with an operational demonstration of the loading of a simulated flight tank with densified propellants. The system will be unique because it uses an integrated refrigeration and storage system (IRAS) to control the state of the fluid. The integrated refrigerator is the critical feature enabling the testing of the following three functions: zero-loss storage and transfer, propellant densification/conditioning, and on-site liquefaction. This paper will discuss the test objectives, the design of the system, and the current status of the installation.

  6. Development and Demonstration of Adanced Tooling Alloys for Molds and Dies

    SciTech Connect

    Kevin M. McHugh; Enrique J. Lavernia

    2006-01-01

    This report summarizes research results in the project Development and Demonstration of Advanced Tooling Alloys for Molds and Dies. Molds, dies and related tooling are used to manufacture most of the plastic and metal products we use every day. Conventional fabrication of molds and dies involves a multiplicity of machining, benching and heat treatment unit operations. This approach is very expensive and time consuming. Rapid Solidifcation Process (RSP) Tooling is a spray-forming technology tailored for producing molds and dies. The appraoch combines rapid solidifcation processing and net-shape materials processing in a single step. An atomized spray of a tool-forming alloy, typically a tool steel, is deposited onto an easy-to-form tool pattern to replicate the pattern's shape and surface features. By so doing, the approach eliminates many machining operations in conventional mold making, significantly reducing cost, lead time and energy. Moreover, rapid solidification creates unique microstructural features by suppressing carbide precipitation and growth, and creating metastable phases. This can result in unique material properties following heat treatment. Spray-formed and aged tool steel dies have exhibited extended life compared to conventional dies in many forming operations such as forging, extrusion and die casting. RSP Tooling technolocy was commercialized with the formation of RSP Tooling, LLC in Solon, Oh.

  7. The regulatory role of Streptomyces coelicolor TamR in central metabolism.

    PubMed

    Huang, Hao; Sivapragasam, Smitha; Grove, Anne

    2015-03-01

    Trans-aconitate methyltransferase regulator (TamR) is a member of the ligand-responsive multiple antibiotic resistance regulator (MarR) family of transcription factors. In Streptomyces coelicolor, TamR regulates transcription of tamR (encoding TamR), tam (encoding trans-aconitate methyltransferase) and sacA (encoding aconitase); up-regulation of these genes promotes metabolic flux through the citric acid cycle. DNA binding by TamR is attenuated and transcriptional derepression is achieved on binding of ligands such as citrate and trans-aconitate to TamR. In the present study, we show that three additional genes are regulated by S. coelicolor TamR. Genes encoding malate synthase (aceB1; SCO6243), malate dehydrogenase (mdh; SCO4827) and isocitrate dehydrogenase (idh; SCO7000) are up-regulated in vivo when citrate and trans-aconitate accumulate, and TamR binds the corresponding gene promoters in vitro, a DNA binding that is attenuated by cognate ligands. Mutations to the TamR binding site attenuate DNA binding in vitro and result in constitutive promoter activity in vivo. The predicted TamR binding sites are highly conserved in the promoters of these genes in Streptomyces species that encode divergent tam-tamR gene pairs, suggesting evolutionary conservation. Like aconitase and trans-aconitate methyltransferase, malate dehydrogenase, isocitrate dehydrogenase and malate synthase are closely related to the citric acid cycle, either catalysing individual reaction steps or, in the case of malate synthase, participating in the glyoxylate cycle to produce malate that enters the citric acid cycle to replenish the intermediate pool. Taken together, our data suggest that TamR plays an important and conserved role in promoting metabolic flux through the citric acid cycle.

  8. A Novel Two-Component System Involved in the Transition to Secondary Metabolism in Streptomyces coelicolor

    PubMed Central

    Rozas, Daniel; Gullón, Sonia; Mellado, Rafael P.

    2012-01-01

    Background Bacterial two-component signal transduction regulatory systems are the major set of signalling proteins frequently mediating responses to changes in the environment. They typically consist of a sensor, a membrane-associated histidine kinase and a cytoplasmic response regulator. The membrane-associated sensor detects the environmental signal or stress, whereas the cytoplasmic regulatory protein controls the cellular response usually by gene transcription modulation. Methodology/PrincipalFindings The Streptomyces coelicolor two genes operon SCO5784-SCO5785 encodes a two-component system, where SCO5784 encodes a histidine-kinase sensor and SCO5785 encodes a response regulator protein. When the expression level of the regulator gene decreases, the antibiotic synthesis and sporulation is delayed temporarily in addition to some ribosomal genes became up regulated, whereas the propagation of the regulatory gene in high copy number results in the earlier synthesis of antibiotics and sporulation, as well as the down regulation of some ribosomal genes and, moreover, in the overproduction of several extracellular proteins. Therefore, this two-component system in S. coelicolor seems to influence various processes characterised by the transition from primary to secondary metabolism, as determined by proteomic and transcriptomic analyses. Conclusions/Significance Propagation of SCO5785 in multicopy enhances the production of antibiotics as well as secretory proteins. In particular, the increase in the expression level of secretory protein encoding genes, either as an artefactual or real effect of the regulator, could be of potential usefulness when using Streptomyces strains as hosts for homologous or heterologous extracellular protein production. PMID:22347508

  9. U.S. Army Environment Center Environmental Quality Technology Development, Demonstration and Transfer Activities

    DTIC Science & Technology

    1994-04-01

    destruction mechanism . Future Plans/Milestones: Hydromill pilot demonstration April 1994 Hydromill pilot report September 1994 Initial demonstration of...soil throughput, amendment mixtures, and two types of composting processes: aerated static pile and mechanically agitated in-vessel composting, were...encouraging. Bench-scale results indicated the need for a pilot-scale demonstration using soil slurry-sequencing batch reactors (SS- SBR ) to determine the

  10. FY 1994 program summary: Office of Technology Development, Office of Research and Development, Office of Demonstration, Testing, and Evaluation

    SciTech Connect

    Not Available

    1994-10-01

    The US Department of Energy (DOE) Office of Environmental Management, formerly the Office of Environmental Restoration and Waste Management (EM), was established in November 1989 as the first step toward correcting contamination problems resulting from nearly 50 years of nuclear weapons production and fuel processing activities. EM consolidates several DOE organizations previously responsible for the handling, treatment, and disposition of radioactive and hazardous waste. Within EM, the Office of Technology Development (OTD/EM-50) is responsible for developing technologies to meet DOE`s goal for environmental restoration. OTD manages an aggressive national program of applied research, development, demonstration, testing, and evaluation (RDDT and E) for environmental cleanup, waste management, and related technologies. The program is designed to resolve major technical issues, to rapidly advanced beyond current technologies for environmental restoration and waste management operations, and to expedite compliance with applicable environmental laws and regulations. This report summarizes Fiscal Year 1994 (FY94) programmatic information, accomplishments, and planned activities relevant to the individual activities within OTD`s RDDT and E.

  11. Developing a Validity Argument through Abductive Reasoning with an Empirical Demonstration of the Latent Class Analysis

    ERIC Educational Resources Information Center

    Wu, Amery D.; Stone, Jake E.; Liu, Yan

    2016-01-01

    This article proposes and demonstrates a methodology for test score validation through abductive reasoning. It describes how abductive reasoning can be utilized in support of the claims made about test score validity. This methodology is demonstrated with a real data example of the Canadian English Language Proficiency Index Program…

  12. Organizing and Conducting Demonstration Projects in Vocational Education. Research and Development Series No. 117.

    ERIC Educational Resources Information Center

    Bina, James V.; Hull, William L.

    This resource book, designed as a planning tool for demonstration project directors, discusses the following six key elements which have application at all three stages of a demonstration project: (1) Objectives, (2) resources, (3) endorsement, (4) communications, (5) evaluation, and (6) outreach. Each of these elements is discussed within the…

  13. Proving the Usefulness of Demonstrations: Using M&M's to Develop Attribute Control Charts

    ERIC Educational Resources Information Center

    Fish, Lynn A.; Braunscheidel, Michael J.

    2012-01-01

    Experiential-based mini-demonstrations are useful to facilitate student learning on a wide variety of topics. The purpose of this teaching brief is two-fold: (1) it outlines a useful mini-demonstration to teach attribute control charting when the sample size is unknown, and (2) adds additional proof that experiential methods positively impact upon…

  14. Content Evaluation and Development of Videotapes Demonstrating Regional Anesthesia Motor Skills

    ERIC Educational Resources Information Center

    Warwick, Pamela M.; Ravin, Mark B.

    1975-01-01

    A study is reported which evaluated the content of three instructional videotapes designed to impart information and to demonstrate regional (spinal, epidural, and caudal) anesthesia motor skills. Pretest-posttest results demonstrated that the tapes successfully met predetermined criteria. Advantages of the method for medical student instruction…

  15. Two genes involved in the phase-variable phi C31 resistance mechanism of Streptomyces coelicolor A3(2).

    PubMed Central

    Bedford, D J; Laity, C; Buttner, M J

    1995-01-01

    The phage growth limitation (Pgl) system of Streptomyces coelicolor confers resistance to phi C31 and its homoimmune phages. The positions of the pgl genes within a 16-kb clone of S. coelicolor DNA were defined by subcloning, insertional inactivation, and deletion mapping. Nucleotide sequencing and functional analysis identified two genes, pglY and pglZ, required for the Pgl+ (phage-resistant) phenotype. pglY and pglZ, which may be translationally coupled, are predicted to encode proteins with M(r)S of 141,000 and 104,000, respectively. Neither protein shows significant similarity to other known proteins, but PglY has a putative ATP/GTP binding motif. The pglY and pglZ genes are cotranscribed from a single promoter which appears to be constitutive and is not induced by phage infection. PMID:7642495

  16. PIAA Coronagraph Development at NASA Ames: High Contrast Laboratory Demonstration at 2 l/D

    NASA Astrophysics Data System (ADS)

    Belikov, Ruslan; Pluzhnik, E.; Witteborn, F. C.; Lynch, D. H.; Greene, T. P.; Zell, P. T.; Balasubramanian, K.; Guyon, O.

    2011-01-01

    Coronagraph technology is advancing and promises to directly image and spectrally characterize extrasolar Earth-like planets in the foreseeable future (such as the 2020 decade) with a telescope as small as 1.5m. A small Explorer-sized telescope can also be launched in the 2010 decade capable of seeing debris disks as small as 10s of zodis and potentially a few large planets. The Phase Induced Amplitude Apodization (PIAA) coronagraph makes such aggressive performance possible. We report on the latest results from a testbed at NASA Ames that is focused on developing and testing the PIAA coronagraph. This laboratory facility was built in 2008 and is designed to be flexible, operated in an actively thermally stabilized air environment, and to complement collaborative efforts at NASA JPL's High Contrast Imaging Testbed. For our wavefront control we are using small Micro-Electro-Mechanical-System deformable mirrors (MEMS DMs), which promise to reduce the size of the beam and overall instrument, a consideration that becomes very important for small telescopes. We describe our lab efforts and results, which include: the operation of our new active thermal control system; the demonstration of 5.4x10-8 (at time of this writing) average raw contrast in a dark zone from 2.0 - 5.2 λ/D in monochromatic light with a refractive PIAA system; preliminary results with an innovative low-cost set of reflective PIAA from JPL; preliminary results with a set of next-generation reflective PIAA built by Tinsley and designed to have the best theoretical broadband performance so far; and finally, an innovative design for a chromatically compensated focal plane occulter that promises to enhance broadband performance by matching the wavelength-dependent inner working angle of coronagraphs such as PIAA.

  17. THE FINE STRUCTURE OF STREPTOMYCES VIOLACEORUBER (S. COELICOLOR)

    PubMed Central

    Glauert, Audrey M.; Hopwood, David A.

    1961-01-01

    A study of thin sections of hyphae of Streptomyces violaceoruber in the electron microscope showed that the structure of the walls and the mode of formation of cross-walls are similar to those of Gram-positive bacteria. A beaded structure was seen in some regions of the wall, and the significance of this observation is discussed in relation to previous studies of the fine structure of bacterial cell walls. Elements of the intracytoplasmic membrane system appear to be involved in the process of cross-wall formation. The walls of the hyphae of the aerial mycelium divide into two layers before the spores are formed, and only the inner component of the wall grows inwards to form the cross-walls and so delimit the spores. The outer component remains intact for a time and acts as a sheath around the developing spores. Finally the sheath breaks and the spores are liberated. This process is contrasted with the formation of endospores in eubacteria. When the spores germinate, the walls of the germ tubes are continuous with those of the spores. PMID:13705984

  18. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1980-01-01

    Presented is a Corridor Demonstration which can be set up in readily accessible areas such as hallways or lobbies. Equipment is listed for a display of three cells (solar cells, fuel cells, and storage cells) which develop electrical energy. (CS)

  19. In Search of the E. coli Compounds that Change the Antibiotic Production Pattern of Streptomyces coelicolor During Inter-species Interaction.

    PubMed

    Mavituna, Ferda; Luti, Khalid Jaber Kadhum; Gu, Lixing

    2016-08-01

    The aim of this work was to investigate the interaction between E.coli and Streptomyces coelicolor A3 (2) for the increased production of undecylprodigiosin and identify the E. coli actives mediating this inter-species interaction. The antibiotics of interest were the red-pigmented undecylprodigiosin and blue-pigmented actinorhodin. Pure cultures of S. coelicolor in a defined medium produced higher concentrations of actinorhodin compared to those of undecylprodigiosin. The latter however, is more important due to its immunosuppressive and antitumor properties. As a strategy to increase undecylprodigiosin production, we added separately, live cells and heat-killed cells of E. coli C600, and the cell-free supernatant of E. coli culture to S. coelicolor cultures in shake flasks. The interaction with live cells of E. coli altered the antibiotic production pattern and undecylprodigiosin production was enhanced by 3.5-fold compared to the pure cultures of S. coelicolor and actinorhodin decreased by 15-fold. The heat-killed cells of E. coli however, had no effect on antibiotic production. In all cases, growth and glucose consumption of S. coelicolor remained almost the same as those observed in the pure culture indicating that the changes in antibiotic production were not due to nutritional stress. Results with cell-free supernatant of E. coli culture indicated that the interaction between S. coelicolor and E. coli was mediated via diffusible molecule(s). Using a set of extraction procedures and agar-well diffusion bioassays, we isolated and preliminarily identified a class of compounds. For the preliminary verification, we added the compound which was the common chemical structural moiety in this class of compounds to the pure S. coelicolor cultures. We observed similar effects on antibiotic production as with the live E. coli cells and their supernatant indicating that this class of compounds secreted by E. coli indeed could act as actives during interspecies

  20. Streptomyces coelicolor encodes a urate-responsive transcriptional regulator with homology to PecS from plant pathogens.

    PubMed

    Huang, Hao; Mackel, Brian J; Grove, Anne

    2013-11-01

    Many transcriptional regulators control gene activity by responding to specific ligands. Members of the multiple-antibiotic resistance regulator (MarR) family of transcriptional regulators feature prominently in this regard, and they frequently function as repressors in the absence of their cognate ligands. Plant pathogens such as Dickeya dadantii encode a MarR homolog named PecS that controls expression of a gene encoding the efflux pump PecM in addition to other virulence genes. We report here that the soil bacterium Streptomyces coelicolor also encodes a PecS homolog (SCO2647) that regulates a pecM gene (SCO2646). S. coelicolor PecS, which exists as a homodimer, binds the intergenic region between pecS and pecM genes with high affinity. Several potential PecS binding sites were found in this intergenic region. The binding of PecS to its target DNA can be efficiently attenuated by the ligand urate, which also quenches the intrinsic fluorescence of PecS, indicating a direct interaction between urate and PecS. In vivo measurement of gene expression showed that activity of pecS and pecM genes is significantly elevated after exposure of S. coelicolor cultures to urate. These results indicate that S. coelicolor PecS responds to the ligand urate by attenuated DNA binding in vitro and upregulation of gene activity in vivo. Since production of urate is associated with generation of reactive oxygen species by xanthine dehydrogenase, we propose that PecS functions under conditions of oxidative stress.

  1. Colonial Differentiation in Streptomyces coelicolor Depends on Translation of a Specific Codon within the adpA Gene

    PubMed Central

    Nguyen, Kien T.; Tenor, Jennifer; Stettler, Hansruedi; Nguyen, Lieu T.; Nguyen, Liem D.; Thompson, Charles J.

    2003-01-01

    We identified adpA as an araC-like regulatory gene needed for colonial morphogenesis in Streptomyces coelicolor and showed that its activity depended on a unique TTA triplet corresponding to the leucyl-tRNA gene (bldA). These findings partially explained the dependence of aerial mycelium formation on a rare tRNA that is postulated to have developmental control functions. PMID:14645292

  2. RNA-Seq and RNA Immunoprecipitation Analyses of the Transcriptome of Streptomyces coelicolor Identify Substrates for RNase III

    PubMed Central

    Gatewood, Marcha L.; Bralley, Patricia; Weil, M. Ryan

    2012-01-01

    RNase III is a key enzyme in the pathways of RNA degradation and processing in bacteria and has been suggested as a global regulator of antibiotic production in Streptomyces coelicolor. Using RNA-Seq, we have examined the transcriptomes of S. coelicolor M145 and an RNase III (rnc)-null mutant of that strain. RNA preparations with reduced levels of structural RNAs were prepared by subtractive hybridization prior to RNA-Seq analysis. We initially identified 7,800 transcripts of known and putative protein-coding genes in M145 and the null mutant, JSE1880, along with transcripts of 21 rRNA genes and 65 tRNA genes. Approximately 3,100 of the protein-coding transcripts were categorized as low-abundance transcripts. For further analysis, we selected those transcripts of known and putative protein-coding genes whose levels changed by ≥2-fold between the two S. coelicolor strains and organized those transcripts into 16 functional categories. We refined our analysis by performing RNA immunoprecipitation of the mRNA preparation from JSE1880 using a mutant RNase III protein that binds to transcripts but does not cleave them. This analysis identified ca. 800 transcripts that were enriched in the RNA immunoprecipitates, including 28 transcripts whose levels also changed by ≥2-fold in the RNA-Seq analysis. We compare our results with those obtained by microarray analysis of the S. coelicolor transcriptome and with studies describing the characterization of small noncoding RNAs. We have also used the RNA immunoprecipitation results to identify new substrates for RNase III cleavage. PMID:22389483

  3. Super Boiler: Packed Media/Transport Membrane Boiler Development and Demonstration

    SciTech Connect

    Liss, William E; Cygan, David F

    2013-04-17

    Gas Technology Institute (GTI) and Cleaver-Brooks developed a new gas-fired steam generation system the Super Boiler for increased energy efficiency, reduced equipment size, and reduced emissions. The system consists of a firetube boiler with a unique staged furnace design, a two-stage burner system with engineered internal recirculation and inter-stage cooling integral to the boiler, unique convective pass design with extended internal surfaces for enhanced heat transfer, and a novel integrated heat recovery system to extract maximum energy from the flue gas. With these combined innovations, the Super Boiler technical goals were set at 94% HHV fuel efficiency, operation on natural gas with <5 ppmv NOx (referenced to 3%O2), and 50% smaller than conventional boilers of similar steam output. To demonstrate these technical goals, the project culminated in the industrial demonstration of this new high-efficiency technology on a 300 HP boiler at Clement Pappas, a juice bottler located in Ontario, California. The Super Boiler combustion system is based on two stage combustion which combines air staging, internal flue gas recirculation, inter-stage cooling, and unique fuel-air mixing technology to achieve low emissions rather than external flue gas recirculation which is most commonly used today. The two-stage combustion provides lower emissions because of the integrated design of the boiler and combustion system which permit precise control of peak flame temperatures in both primary and secondary stages of combustion. To reduce equipment size, the Super Boiler's dual furnace design increases radiant heat transfer to the furnace walls, allowing shorter overall furnace length, and also employs convective tubes with extended surfaces that increase heat transfer by up to 18-fold compared to conventional bare tubes. In this way, a two-pass boiler can achieve the same efficiency as a traditional three or four-pass firetube boiler design. The Super Boiler is consequently up to

  4. Global negative regulation of Streptomyces coelicolor antibiotic synthesis mediated by an absA-encoded putative signal transduction system.

    PubMed Central

    Brian, P; Riggle, P J; Santos, R A; Champness, W C

    1996-01-01

    Streptomycete antibiotic synthesis is coupled to morphological differentiation such that antibiotics are produced as a colony sporulates. Streptomyces coelicolor produces several structurally and genetically distinct antibiotics. The S. coelicolor absA locus was defined by four UV-induced mutations that globally blocked antibiotic biosynthesis without blocking morphological differentiation. We show that the absA locus encodes a putative eubacterial two-component sensor kinase-response regulator system. All four mutations lie within a single open reading frame, designated absA1, which is predicted to encode a sensor histidine kinase. A second gene downstream of absA1, absA2, is predicted to encode the cognate response regulator. In marked contrast to the antibiotic-deficient phenotype of the previously described absA mutants, the phenotype caused by disruption mutations in the absA locus is precocious hyperproduction of the antibiotics actinorhodin and undecylprodigiosin. Precocious hyperproduction of these antibiotics is correlated with premature expression of XylE activity in a transcriptional fusion to an actinorhodin biosynthetic gene. We propose that the absA locus encodes a signal transduction mechanism that negatively regulates synthesis of the multiple antibiotics produced by S. coelicolor. PMID:8655502

  5. Medium engineering for enhanced production of undecylprodigiosin antibiotic in Streptomyces coelicolor using oil palm biomass hydrolysate as a carbon source.

    PubMed

    Bhatia, Shashi Kant; Lee, Bo-Rahm; Sathiyanarayanan, Ganesan; Song, Hun-Seok; Kim, Junyoung; Jeon, Jong-Min; Kim, Jung-Ho; Park, Sung-Hee; Yu, Ju-Hyun; Park, Kyungmoon; Yang, Yung-Hun

    2016-10-01

    In this study, a biosugar obtained from empty fruit bunch (EFB) of oil palm by hot water treatment and subsequent enzymatic saccharification was used for undecylprodigiosin production, using Streptomyces coelicolor. Furfural is a major inhibitor present in EFB hydrolysate (EFBH), having a minimum inhibitory concentration (MIC) of 1.9mM, and it reduces utilization of glucose (27%), xylose (59%), inhibits mycelium formation, and affects antibiotic production. Interestingly, furfural was found to be a good activator of undecylprodigiosin production in S. coelicolor, which enhanced undecylprodigiosin production by up to 52%. Optimization by mixture analysis resulted in a synthetic medium containing glucose:furfural:ACN:DMSO (1%, 2mM, 0.2% and 0.3%, respectively). Finally, S. coelicolor was cultured in a fermenter in minimal medium with EFBH as a carbon source and addition of the components described above. This yielded 4.2μg/mgdcw undecylprodigiosin, which was 3.2-fold higher compared to that in un-optimized medium.

  6. DEVELOPMENT AND DEMONSTRATION OF NOVEL LOW-NOx BURNERS IN THE STEEL INDUSTRY

    SciTech Connect

    Cygan, David

    2006-12-28

    Gas Technology Institute (GTI), together with Hamworthy Peabody Combustion Incorporated (formerly Peabody Engineering Corporation), the University of Utah, and Far West Electrochemical have developed and demonstrated an innovative combustion system suitable for natural gas and coke-oven gas firing within the steel industry. The combustion system is a simple, low-cost, energy-efficient burner that can reduce NOx by more than 75%. The U.S. steel industry needs to address NOx control at its steelmaking facilities. A significant part of NOx emissions comes from gas-fired boilers. In steel plants, byproduct gases – blast furnace gas (BFG) and coke-oven gas (COG) – are widely used together with natural gas to fire furnaces and boilers. In steel plants, natural gas can be fired together with BFG and COG, but, typically, the addition of natural gas raises NOx emissions, which can already be high because of residual fuel-bound nitrogen in COG. The Project Team has applied its expertise in low-NOx burners to lower NOx levels for these applications by combining advanced burner geometry and combustion staging with control strategies tailored to mixtures of natural gas and byproduct fuel gases. These methods reduce all varieties of NOx – thermal NOx produced by high flame temperatures, prompt NOx produced by complex chain reactions involving radical hydrocarbon species and NOx from fuel-bound nitrogen compounds such as ammonia found in COG. The Project Team has expanded GTI’s highly successful low-NOx forced internal recirculation (FIR) burner, previously developed for natural gas-fired boilers, into facilities that utilize BFG and COG. For natural gas firing, these burners have been shown to reduce NOx emissions from typical uncontrolled levels of 80-100 vppm to single-digit levels (9 vppm). This is done without the energy efficiency penalties incurred by alternative NOx control methods, such as external flue gas recirculation (FGR), water injection, and selective non

  7. Novel High-Voltage, High-Power Piezoelectric Transformer Developed and Demonstrated for Space Communications Applications

    NASA Technical Reports Server (NTRS)

    Carazo, Alfredo V.; Wintucky, Edwin G.

    2004-01-01

    Improvements in individual piezoelectric transformer (PT) performance and the combination of these PTs in a unique modular topology under a Phase I contract with the NASA Glenn Research Center have enabled for the first time the simultaneous achievement of both high voltage and high power at much higher levels than previously obtained with any PT. Feasibility was demonstrated by a prototype transformer (called a Tap-Soner), which is shown in the preceding photograph as part of a direct-current to direct-current (dc-dc) converter having two outputs rated at 1.5 kV/5 W and 4.5 kV/20 W. The power density of 3.5 W/cm3 is significantly lower than for magnetic transformers with the same voltage and power output. This development, which is being done under a Small Business Innovation Research (SBIR) contract by Face Electronics, LC (Norfolk, VA), is based on improvements in the materials and design of Face's basic patented Transoner-T3 PT, shown in the left in the following figure. The T3 PT is most simply described as a resonant multilayer transducer where electrical energy at the input section is efficiently mechanically coupled to the output section, which then vibrates in a fundamental longitudinal mode to generate a high gain in voltage. The piezoelectric material used is a modified lead-zirconium-titanate-based ceramic. One of the significant improvements in PT design was the incorporation of a symmetrical double input layer, shown on the right in the following figure, which eliminated the lossy bending vibration modes characteristic of a single input layer. The performance of the improved PT was optimized to 1.5 kV/5 W. The next step was devising a way to combine the individual PTs in a modular circuit topology needed to achieve the desired high voltage and power output. Since the optimum performance of the individual PT occurs at resonance, the most efficient operation of the modular transformer was achieved by using a separate drive circuit for each PT. The

  8. Evaluation of waste treatment technologies by LLWDDD (Low-Level Waste Disposal Development and Demonstration) Programs

    SciTech Connect

    Kennerly, J.M.; Williams, L.C.; Dole, L.R.; Genung, R.K.

    1987-01-01

    Waste treatments are divided into four categories: (1) volume reduction; (2) conditioning to improve waste form performance; (3) segregation to achieve waste reduction; and (4) separation to remove radioactive (or hazardous) constituents. Two waste treatment demonstrations are described. In the first, volume reduction by mechanical means was achieved during the supercompaction of 300 55-gal drums of solid waste at ORNL. In the second demonstration, conditioning of waste through immobilization and packaging to improve the performance of the waste form is being evaluated. The final section of this paper describes potential scenarios for the management of uranium-contaminated wastes at the Y-12 Plant in Oak Ridge and emphasizes where demonstrations of treatment technology will be needed to implement the scenarios. Separation and thermal treatment are identified as the principal means for treating these wastes. 15 figs.

  9. Development of Supersonic Vehicle for Demonstration of a Precooled Turbojet Engine

    NASA Astrophysics Data System (ADS)

    Sawai, Shujiro; Fujita, Kazuhisa; Kobayashi, Hiroaki; Sakai, Shin'ichiro; Bando, Nobutaka; Kadooka, Shouhei; Tsuboi, Nobuyuki; Miyaji, Koji; Uchiyama, Taku; Hashimoto, Tatsuaki

    JAXA is developing Mach 5 hypersonic turbojet engine technology that can be applied in a future hypersonic transport. Now, Jet Engine Technology Research Center of JAXA conducts the experimental study using a 1 / 10 scale-model engine. In parallel to engine development activities, a new supersonic flight-testing vehicle for the hypersonic turbojet engine is under development since 2004. In this paper, the system configuration of the flight-testing vehicle is outlined and development status is reported.

  10. Development and Demonstration of the Open Automated Demand Response Standard for the Residential Sector

    SciTech Connect

    Herter, Karen; Rasin, Josh; Perry, Tim

    2009-11-30

    The goal of this study was to demonstrate a demand response system that can signal nearly every customer in all sectors through the integration of two widely available and non- proprietary communications technologies--Open Automated Demand Response (OpenADR) over lnternet protocol and Utility Messaging Channel (UMC) over FM radio. The outcomes of this project were as follows: (1) a software bridge to allow translation of pricing signals from OpenADR to UMC; and (2) a portable demonstration unit with an lnternet-connected notebook computer, a portfolio of DR-enabling technologies, and a model home. The demonstration unit provides visitors the opportunity to send electricity-pricing information over the lnternet (through OpenADR and UMC) and then watch as the model appliances and lighting respond to the signals. The integration of OpenADR and UMC completed and demonstrated in this study enables utilities to send hourly or sub-hourly electricity pricing information simultaneously to the residential, commercial and industrial sectors.

  11. Scenarios for Benefits Analysis of Energy Research, Development,Demonstration and Deployment

    SciTech Connect

    Gumerman, Etan; Marnay, Chris

    2005-09-07

    For at least the last decade, evaluation of the benefits of research, development, demonstration, and deployment (RD3) by the U.S. Department of Energy has been conducted using deterministic forecasts that unrealistically presume we can precisely foresee our future 10, 25,or even 50 years hence. This effort tries, in a modest way, to begin a process of recognition that the reality of our energy future is rather one rife with uncertainty. The National Energy Modeling System (NEMS) is used by the Department of Energy's Office of Energy Efficiency and Renewable Energy (EE) and Fossil Energy (FE) for their RD3 benefits evaluation. In order to begin scoping out the uncertainty in these deterministic forecasts, EE and FE designed two futures that differ significantly from the basic NEMS forecast. A High Fuel Price Scenario and a Carbon Cap Scenario were envisioned to forecast alternative futures and the associated benefits. Ernest Orlando Lawrence Berkeley National Laboratory (LBNL) implemented these scenarios into its version of NEMS,NEMS-LBNL, in late 2004, and the Energy Information Agency created six scenarios for FE in early 2005. The creation and implementation of the EE-FE scenarios are explained in this report. Both a Carbon Cap Scenario and a High Fuel Price Scenarios were implemented into the NEMS-LBNL. EIA subsequently modeled similar scenarios using NEMS. While the EIA and LBNL implementations were in some ways rather different, their forecasts do not significantly diverge. Compared to the Reference Scenario, the High Fuel Price Scenario reduces energy consumption by 4 percent in 2025, while in the EIA fuel price scenario (known as Scenario 4) reduction from its corresponding reference scenario (known as Scenario 0) in 2025 is marginal. Nonetheless, the 4 percent demand reduction does not lead to other cascading effects that would significantly differentiate the two scenarios. The LBNL and EIA carbon scenarios were mostly identical. The only major difference

  12. Engineering Model Propellant Feed System Development for an Iodine Hall Thruster Demonstration Mission

    NASA Technical Reports Server (NTRS)

    Polzin, Kurt A.

    2016-01-01

    CUBESATS are relatively new spacecraft platforms that are typically deployed from a launch vehicle as a secondary payload, providing low-cost access to space for a wide range of end-users. These satellites are comprised of building blocks having dimensions of 10x10x10 cu cm and a mass of 1.33 kg (a 1-U size). While providing low-cost access to space, a major operational limitation is the lack of a propulsion system that can fit within a CubeSat and is capable of executing high (Delta)v maneuvers. This makes it difficult to use CubeSats on missions requiring certain types of maneuvers (i.e. formation flying, spacecraft rendezvous). Recently, work has been performed investigating the use of iodine as a propellant for Hall-effect thrusters (HETs) 2 that could subsequently be used to provide a high specific impulse path to CubeSat propulsion. 3, 4 Iodine stores as a dense solid at very low pressures, making it acceptable as a propellant on a secondary payload. It has exceptionally high ?Isp (density times specific impulse), making it an enabling technology for small satellite near-term applications and providing the potential for systems-level advantages over mid-term high power electric propulsion options. Iodine flow can also be thermally regulated, subliming at relatively low temperature (< 100 C) to yield I2 vapor at or below 50 torr. At low power, the measured performance of an iodine-fed HET is very similar to that of a state-of-the-art xenon-fed thruster. Just as importantly, the current-voltage discharge characteristics of low power iodine-fed and xenon-fed thrusters are remarkably similar, potentially reducing development and qualifications costs by making it possible to use an already-qualified xenon-HET PPU in an iodine-fed system. Finally, a cold surface can be installed in a vacuum test chamber on which expended iodine propellant can deposit. In addition, the temperature doesn't have to be extremely cold to maintain a low vapor pressure in the vacuum

  13. Development and Evaluation of Sensor Concepts for Ageless Aerospace Vehicles: Report 4 - Phase 1 Implementation of the Concept Demonstrator

    NASA Technical Reports Server (NTRS)

    Abbott, David; Batten, Adam; Carpenter, David; Dunlop, John; Edwards, Graeme; Farmer, Tony; Gaffney, Bruce; Hedley, Mark; Hoschke, Nigel; Isaacs, Peter; Johnson, Mark; Lewis, Chris; Murdoch, Alex; Poulton, Geoff; Price, Don; Prokopenko, Mikhail; Rees, David; Scott, Andrew; Seneviratne, Sarath; Valencia, Philip; Wang, Peter; Whitnall, Denis

    2008-01-01

    This report describes the first phase of the implementation of the Concept Demonstrator. The Concept Demonstrator system is a powerful and flexible experimental test-bed platform for developing sensors, communications systems, and multi-agent based algorithms for an intelligent vehicle health monitoring system for deployment in aerospace vehicles. The Concept Demonstrator contains sensors and processing hardware distributed throughout the structure, and uses multi-agent algorithms to characterize impacts and determine an appropriate response to these impacts.

  14. Biochemical studies of the multicopper oxidase (small laccase) from Streptomyces coelicolor using bioactive phytochemicals and site-directed mutagenesis

    PubMed Central

    Sherif, Mohammed; Waung, Debbie; Korbeci, Bihter; Mavisakalyan, Valentina; Flick, Robert; Brown, Greg; Abou-Zaid, Mamdouh; Yakunin, Alexander F; Master, Emma R

    2013-01-01

    Summary Multicopper oxidases can act on a broad spectrum of phenolic and non-phenolic compounds. These enzymes include laccases, which are widely distributed in plants and fungi, and were more recently identified in bacteria. Here, we present the results of biochemical and mutational studies of small laccase (SLAC), a multicopper oxidase from Streptomyces coelicolor (SCO6712). In addition to typical laccase substrates, SLAC was tested using phenolic compounds that exhibit antioxidant activity. SLAC showed oxidase activity against 12 of 23 substrates tested, including caffeic acid, ferulic acid, resveratrol, quercetin, morin, kaempferol and myricetin. The kinetic parameters of SLAC were determined for 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid), 2,6-dimethoxyphenol, quercetin, morin and myricetin, and maximum reaction rates were observed with myricetin, where kcat and Km values at 60°C were 8.1 (± 0.8) s−1 and 0.9 (± 0.3) mM respectively. SLAC had a broad pH optimum for activity (between pH 4 and 8) and temperature optimum at 60–70°C. It demonstrated remarkable thermostability with a half-life of over 10 h at 80°C and over 7 h at 90°C. Site-directed mutagenesis revealed 17 amino acid residues important for SLAC activity including the 10 His residues involved in copper coordination. Most notably, the Y229A and Y230A mutant proteins showed over 10-fold increase in activity compared with the wild-type SLAC, which was correlated to higher copper incorporation, while kinetic analyses with S929A predicts localization of this residue near the meta-position of aromatic substrates. Funding Information Funding for this research was provided by the Government of Ontario for the project ‘FFABnet: Functionalized Fibre and Biochemicals’ (ORF-RE-05-005), and the Natural Sciences and Engineering Research Council of Canada. PMID:23815400

  15. Research development and demonstration of a fuel cell/battery powered bus system. Annual report, January 1--December 31, 1994

    SciTech Connect

    Wimmer, R.

    1995-01-01

    This report describes the progress in the Georgetown University research, development and demonstration project of a fuel cell/battery powered bus system. The topics addressed in the report include demonstrations, vehicle design and application analysis, technology transfer activities, coordination and monitoring of system design and integration contractor, fuel cell bus test program, current problems, work planned, and manpower, cost and schedule reports.

  16. Development of Criteria to Measure the Extent of Implementation and the Effectiveness of Demonstration in Vocational Education. Case Studies.

    ERIC Educational Resources Information Center

    Development Associates, Inc., Arlington, VA.

    This document presents case studies of eight demonstration projects selected to assess the practicality and feasibility of a monograph for improving and measuring the impact of vocational education demonstration projects. An introduction provides background of the project that developed that monograph and information on the case studies, including…

  17. A small scale accelerator driven subcritical assembly development and demonstration experiment at LAMPF

    SciTech Connect

    Wender, S. A.; Venneri, F.; Bowman, C. D.; Arthur, E. D.; Heighway, E.; Beard, C. A.; Bracht, R. R.; Buksa, J. J.; Chavez, W.; DeVolder, B. G.; Park, J. J.; Parker, R. B.; Pillai, C.; Pitcher, E.; Potter, R. C.; Reid, R. S.; Russell, G. J.; Trujillo, D. A.; Weinacht, D. J.; Wilson, W. B.

    1995-09-15

    A small scale experiment is described that will demonstrate many of the aspects of accelerator-driven transmutation technology. This experiment uses the high-power proton beam from the Los Alamos Meson Physics Facility accelerator and will be located in the Area-A experimental hall. Beam currents of up to 1 mA will be used to produce neutrons with a molten lead target. The target is surrounded by a molten salt and graphite moderator blanket. Fissionable material can be added to the molten salt to demonstrate plutonium burning or transmutation of commercial spent fuel or energy production from thorium. The experiment will be operated at power levels up to 5 MWt.

  18. The development of the human exploration demonstration project (HEDP), a planetary systems testbed

    NASA Technical Reports Server (NTRS)

    Chevers, Edward S.; Korsmeyer, David J.

    1993-01-01

    The Human Exploration Demonstration Project (HEDP) is an ongoing task at the National Aeronautics and Space Administration's Ames Research Center to address the advanced technology requirements necessary to implement an integrated working and living environment for a planetary surface habitat. The integrated environment will consist of life support systems, physiological monitoring of project crew, a virtual environment workstation, and centralized data acquisition and habitat systems health monitoring. There will be several robotic systems on a simulated planetary landscape external to the habitat environment to provide representative work loads for the crew. This paper describes the status of the HEDP after one year, the major facilities composing the HEDP, the project's role as an Ames Research Center testbed, and the types of demonstration scenarios that will be run to showcase the technologies.

  19. XV-15 Tilt Rotor fly-by-wire collective control demonstrator development specifications

    NASA Technical Reports Server (NTRS)

    Meuleners, R. J.

    1981-01-01

    A fly by wire system in the collective control system for XV-15 Tilt Rotor Research Aircraft was evaluated. The collective control system was selected because it requires a system tracking accuracy between right and left rotors of approximately 0.1%. The performance characteristics of the collectors axel provide typical axis control response data. The demonstrator is bread boarded as a dual system instead of the triplex system.

  20. Development and Demonstration of a Biomass Boiler for Food Processing Applications

    SciTech Connect

    2009-02-01

    Burns & McDonnell Engineering Company, in collaboration with Frito-Lay, Inc., Oak Ridge National Laboratory, CPL Systems, Inc., Alpha Boilers, and Kansas State University will demonstrate use of a biomass boiler in the food processing industry. The 60,000 lb/hr innovative biomass boiler system utilizing a combination of wood waste and tire-derived fuel (TDF) waste will offset all natural gas consumption at Frito-Lay's Topeka, Kansas, processing facility.

  1. ςBldN, an Extracytoplasmic Function RNA Polymerase Sigma Factor Required for Aerial Mycelium Formation in Streptomyces coelicolor A3(2)

    PubMed Central

    Bibb, Maureen J.; Molle, Virginie; Buttner, Mark J.

    2000-01-01

    Sporulation mutants of Streptomyces coelicolor appear white because they are defective in the synthesis of the gray polyketide spore pigment, and such white (whi) mutants have been used to define 13 sporulation loci. whiN, one of five new whi loci identified in a recent screen of NTG (N-methyl-N′-nitro-N-nitrosoguanidine)-induced whi strains (N. J. Ryding et al., J. Bacteriol. 181:5419–5425, 1999), was defined by two mutants, R112 and R650. R650 produced frequent spores that were longer than those of the wild type. In contrast, R112 produced long, straight, undifferentiated hyphae, although rare spore chains were observed, sometimes showing highly irregular septum placement. Subcloning and sequencing showed that whiN encodes a member of the extracytoplasmic function subfamily of RNA polymerase sigma factors and that the sigma factor has an unusual N-terminal extension of approximately 86 residues that is not present in other sigma factors. A constructed whiN null mutant failed to form aerial mycelium (the “bald” phenotype) and, as a consequence, whiN was renamed bldN. This observation was not totally unexpected because, on some media, the R112 point mutant produced substantially less aerial mycelium than its parent, M145. The bldN null mutant did not fit simply into the extracellular signaling cascade proposed for S. coelicolor bld mutants. Expression of bldN was analyzed during colony development in wild-type and aerial mycelium-deficient bld strains. bldN was transcribed from a single promoter, bldNp. bldN transcription was developmentally regulated, commencing approximately at the time of aerial mycelium formation, and depended on bldG and bldH, but not on bldA, bldB, bldC, bldF, bldK, or bldJ or on bldN itself. Transcription from the p1 promoter of the response-regulator gene bldM depended on bldN in vivo, and the bldMp1 promoter was shown to be a direct biochemical target for ςBldN holoenzyme in vitro. PMID:10913095

  2. sigma(BldN), an extracytoplasmic function RNA polymerase sigma factor required for aerial mycelium formation in Streptomyces coelicolor A3(2).

    PubMed

    Bibb, M J; Molle, V; Buttner, M J

    2000-08-01

    Sporulation mutants of Streptomyces coelicolor appear white because they are defective in the synthesis of the gray polyketide spore pigment, and such white (whi) mutants have been used to define 13 sporulation loci. whiN, one of five new whi loci identified in a recent screen of NTG (N-methyl-N'-nitro-N-nitrosoguanidine)-induced whi strains (N. J. Ryding et al., J. Bacteriol. 181:5419-5425, 1999), was defined by two mutants, R112 and R650. R650 produced frequent spores that were longer than those of the wild type. In contrast, R112 produced long, straight, undifferentiated hyphae, although rare spore chains were observed, sometimes showing highly irregular septum placement. Subcloning and sequencing showed that whiN encodes a member of the extracytoplasmic function subfamily of RNA polymerase sigma factors and that the sigma factor has an unusual N-terminal extension of approximately 86 residues that is not present in other sigma factors. A constructed whiN null mutant failed to form aerial mycelium (the "bald" phenotype) and, as a consequence, whiN was renamed bldN. This observation was not totally unexpected because, on some media, the R112 point mutant produced substantially less aerial mycelium than its parent, M145. The bldN null mutant did not fit simply into the extracellular signaling cascade proposed for S. coelicolor bld mutants. Expression of bldN was analyzed during colony development in wild-type and aerial mycelium-deficient bld strains. bldN was transcribed from a single promoter, bldNp. bldN transcription was developmentally regulated, commencing approximately at the time of aerial mycelium formation, and depended on bldG and bldH, but not on bldA, bldB, bldC, bldF, bldK, or bldJ or on bldN itself. Transcription from the p1 promoter of the response-regulator gene bldM depended on bldN in vivo, and the bldMp1 promoter was shown to be a direct biochemical target for sigma(BldN) holoenzyme in vitro.

  3. A Complex Insertion Sequence Cluster at a Point of Interaction between the Linear Plasmid SCP1 and the Linear Chromosome of Streptomyces coelicolor A3(2)

    PubMed Central

    Yamasaki, Masayuki; Miyashita, Kiyotaka; Cullum, John; Kinashi, Haruyasu

    2000-01-01

    The giant linear plasmid SCP1 can integrate into the central region of the linear chromosome of Streptomyces coelicolor A3(2). Nucleotide sequence analysis around the target site for SCP1 integration in strain M145 identified a total of five copies of four insertion sequences (ISs) in a 6.5-kb DNA stretch. Three of the four (IS468, IS469, and IS470) are new IS elements, and the other is IS466. All of these elements contain one open reading frame which encodes a transposase-like protein. Two copies of IS468 (IS468A and -B) are tandemly aligned at the left end of the cluster. Following these, IS469 and IS466 are located in a tail-to-tail orientation with 69.3% identity to each other. IS470 is located at the right end of the cluster. The activities of IS466 and IS468 were demonstrated by transposition experiments and sequence comparison of several copies, respectively. PMID:10809688

  4. The phage growth limitation system in Streptomyces coelicolor A(3)2 is a toxin/antitoxin system, comprising enzymes with DNA methyltransferase, protein kinase and ATPase activity.

    PubMed

    Hoskisson, Paul A; Sumby, Paul; Smith, Margaret C M

    2015-03-01

    The phage growth limitation system of Streptomyces coelicolor A3(2) is an unusual bacteriophage defence mechanism. Progeny ϕC31 phage from an initial infection are thought to be modified such that subsequent infections are attenuated in a Pgl(+) host but normal in a Pgl(-) strain. Earlier work identified four genes required for phage resistance by Pgl. Here we demonstrate that Pgl is an elaborate and novel phage restriction system that, in part, comprises a toxin/antitoxin system where PglX, a DNA methyltransferase is toxic in the absence of a functional PglZ. In addition, the ATPase activity of PglY and a protein kinase activity in PglW are shown to be essential for phage resistance by Pgl. We conclude that on infection of a Pgl(+) cell by bacteriophage ϕC31, PglW transduces a signal, probably via phosphorylation, to other Pgl proteins resulting in the activation of the DNA methyltransferase, PglX and this leads to phage restriction.

  5. Molecular characterization of SCO0765 as a cellotriose releasing endo-β-1,4-cellulase from Streptomyces coelicolor A(3).

    PubMed

    Hong, Joo-Bin; Dhakshnamoorthy, Vijayalakshmi; Lee, Chang-Ro

    2016-09-01

    The sco0765 gene was annotated as a glycosyl hydrolase family 5 endoglucanase from the genomic sequence of Streptomyces coelicolor A3(2) and consisted of 2,241 bp encoding a polypeptide of 747 amino acids (molecular weight of 80.5 kDa) with a 29-amino acid signal peptide for secretion. The SCO0765 recombinant protein was heterogeneously over-expressed in Streptomyces lividans TK24 under the control of a strong ermE* promoter. The purified SCO0765 protein showed the expected molecular weight of the mature form (718 aa, 77.6 kDa) on sodium dodecyl sulfate-polyacryl amide gel electrophoresis. SCO0765 showed high activity toward β-glucan and carboxymethyl cellulose (CMC) and negligible activity to Avicel, xylan, and xyloglucan. The SCO0765 cellulase had a maximum activity at pH 6.0 and 40°C toward CMC and at pH 9.0 and 50-60°C toward β-glucan. Thin layer chromatography of the hydrolyzed products of CMC and β-glucan by SCO0765 gave cellotriose as the major product and cellotetraose, cellopentaose, and longer oligosaccharides as the minor products. These results clearly demonstrate that SCO0765 is an endo-β-1,4-cellulase, hydrolyzing the β-1,4 glycosidic bond of cellulose into cellotriose.

  6. Molecular and Functional Analyses of the Gene (eshA) Encoding the 52-Kilodalton Protein of Streptomyces coelicolor A3(2) Required for Antibiotic Production

    PubMed Central

    Kawamoto, Shinichi; Watanabe, Masakatsu; Saito, Natsumi; Hesketh, Andrew; Vachalova, Katerina; Matsubara, Keiko; Ochi, Kozo

    2001-01-01

    Analysis of proteins recovered in the S100 precipitate fraction of Streptomyces griseus after ultracentrifugation led to the identification of a 52-kDa protein which is produced during the late growth phase. The gene (eshA) which codes for this protein was cloned from S. griseus, and then its homologue was cloned from Streptomyces coelicolor A3(2). The protein was deduced to be 471 amino acids in length. The protein EshA is characterized by a central region that shows homology to the eukaryotic-type cyclic nucleotide-binding domains. Significant homology was also found to MMPI in Mycobacterium leprae, a major antigenic protein to humans. The eshA gene mapped near the chromosome end and was not essential for viability, as demonstrated by gene disruption experiments, but its disruption resulted in the abolishment of an antibiotic (actinorhodin but not undecylprodigiosin) production. Aerial mycelium was produced as abundantly as by the parent strain. Expression analysis of the EshA protein by Western blotting revealed that EshA is present only in late-growth-phase cells. The eshA gene was transcribed just preceding intracellular accumulation of the EshA protein, as determined by S1 nuclease protection, indicating that EshA expression is regulated at the transcription level. The expression of EshA was unaffected by introduction of the relA mutation, which blocks ppGpp synthesis. PMID:11567001

  7. The Streptomyces coelicolor lipoate-protein ligase is a circularly permuted version of the Escherichia coli enzyme composed of discrete interacting domains.

    PubMed

    Cao, Xinyun; Cronan, John E

    2015-03-13

    Lipoate-protein ligases are used to scavenge lipoic acid from the environment and attach the coenzyme to its cognate proteins, which are generally the E2 components of the 2-oxoacid dehydrogenases. The enzymes use ATP to activate lipoate to its adenylate, lipoyl-AMP, which remains tightly bound in the active site. This mixed anhydride is attacked by the ϵ-amino group of a specific lysine present on a highly conserved acceptor protein domain, resulting in the amide-linked coenzyme. The Streptomyces coelicolor genome encodes only a single putative lipoate ligase. However, this protein had only low sequence identity (<25%) to the lipoate ligases of demonstrated activity and appears to be a circularly permuted version of the known lipoate ligase proteins in that the canonical C-terminal domain seems to have been transposed to the N terminus. We tested the activity of this protein both by in vivo complementation of an Escherichia coli ligase-deficient strain and by in vitro assays. Moreover, when the domains were rearranged into a protein that mimicked the arrangement found in the canonical lipoate ligases, the enzyme retained complementation activity. Finally, when the two domains were separated into two proteins, both domain-containing proteins were required for complementation and catalysis of the overall ligase reaction in vitro. However, only the large domain-containing protein was required for transfer of lipoate from the lipoyl-AMP intermediate to the acceptor proteins, whereas both domain-containing proteins were required to form lipoyl-AMP.

  8. Affordable Development and Demonstration of a Small NTR engine and Stage: A Preliminary NASA, DOE, and Industry Assessment

    NASA Technical Reports Server (NTRS)

    Borowski, S. K.; Sefcik, R. J.; Fittje, J. E.; McCurdy, D. R.; Qualls, A. L.; Schnitzler, B. G; Werner, J.; Weitzberg, A.; Joyner, C. R.

    2015-01-01

    In FY'11, Nuclear Thermal Propulsion (NTP) was identified as a key propulsion option under the Advanced In-Space Propulsion (AISP) component of NASA's Exploration Technology Development and Demonstration (ETDD) program A strategy was outlined by GRC and NASA HQ that included 2 key elements -"Foundational Technology Development" followed by specific "Technology Demonstration" projects. The "Technology Demonstration "element proposed ground technology demonstration (GTD) testing in the early 2020's, followed by a flight technology demonstration (FTD) mission by approx. 2025. In order to reduce development costs, the demonstration projects would focus on developing a small, low thrust (approx. 7.5 -16.5 klb(f)) engine that utilizes a "common" fuel element design scalable to the higher thrust (approx. 25 klb(f)) engines used in NASA's Mars DRA 5.0 study(NASA-SP-2009-566). Besides reducing development costs and allowing utilization of existing, flight proven engine hard-ware (e.g., hydrogen pumps and nozzles), small, lower thrust ground and flight demonstration engines can validate the technology and offer improved capability -increased payloads and decreased transit times -valued for robotic science missions identified in NASA's Decadal Study.

  9. Books and Becoming Good: Demonstrating Aristotle's Theory of Moral Development in the Act of Reading

    ERIC Educational Resources Information Center

    Cain, Amanda

    2005-01-01

    In the Nicomachean ethics, Aristotle sets down a scattered and fractional account of the development of moral virtue within young people. Philosopher Martha Nussbaum defends Aristotle's neglect of a systematic account of moral development and argues that more complex expressions of character-building, such as learning to expose oneself to proper…

  10. Research, development and demonstration of nickel-zinc batteries for electric vehicle propulsion. Annual report, 1979

    SciTech Connect

    Not Available

    1980-06-01

    Activities in a program to develop a Ni/Zn battery for electric vehicle propulsion are reported. Aspects discussed include battery design and development, nickel cathode study, and basic electrochemistry. A number of engineering drawings are supplied. 61 figures, 11 tables. (RWR)

  11. DEMONSTRATION BULLETIN: COLD TOP EX-SITU VITRIFICATION PROCESS - GEOTECH DEVELOPMENT CORPORATION

    EPA Science Inventory

    The Cold Top Vitrification process, developed by Geotech Development Corporation, is an ex-situ, submerged-electrode, resistance-melting technology. The technology is designed to transform heavy metal contaminated soil into a glassy, amorphous, non-leachable mass composed of inte...

  12. Research, development and demonstration of nickel-zinc batteries for electric vehicle propulsion. Annual report, 1979

    SciTech Connect

    Not Available

    1980-06-01

    Progress achieved under ANL Contract No. 31-109-38-4248 from 16 August 1978 to 16 August 1979 is reported. The first segment of the overall program, component development, consists of four basic tasks proceeding in parallel: nickel electrode development, zinc electrode development, separator development, and sealed cell development. Each of these tasks is reported herein on a self-contained basis. System engineering is the second major subdivision of the effort. It includes the design and testing of all cells, the investigation of charge control devices and techniques, and the complete analysis of all cells for failure modes. It also encompasses the accelerated testing of 20-Ah cells. To date, large numbers of these cells (incorporating separator variations, active material additives and internal design variations) have been subjected to this type of testing. 48 figures, 47 tables. (RWR)

  13. Development experience with a simple expert system demonstrator for pilot emergency procedures

    NASA Technical Reports Server (NTRS)

    Vannorman, M.; Mackall, D. A.

    1986-01-01

    Expert system techniques, a major application area of artificial intelligence (AI), are examined in the development of pilot associate to handle aircraft emergency procedures. The term pilot associate is used to describe research involving expert systems that can assist the pilot in the cockpit. The development of expert systems for the electrical system and flight control system emergency procedures are discussed. A simple, high-level expert system provides the means to choose which knowledge domain is needed. The expert systems were developed on a low-cost, FORTH-based package, using a personal computer.

  14. Development of the Global Ozone Lidar Demonstrator (GOLD) Instrument for Deployment on the NASA Global Hawk

    NASA Technical Reports Server (NTRS)

    Hair, Jonathan W.; Browell, Edward V.; McGee, Thomas; Butler, Carolyn; Fenn, Marta; Os,ao (. Sued); Notari, Anthony; Collins, James; Cleckner, Craig; Hostetler, Chris

    2010-01-01

    A compact ozone (O3) and aerosol lidar system is being developed for conducting global atmospheric investigations from the NASA Global Hawk Uninhabited Aerial Vehicle (UAV) and for enabling the development and test of a space-based O3 and aerosol lidar. GOLD incorporates advanced technologies and designs to produce a compact, autonomously operating O3 and aerosol Differential Absorption Lidar (DIAL) system for a UAV platform. The GOLD system leverages advanced Nd:YAG and optical parametric oscillator laser technologies and receiver optics, detectors, and electronics. Significant progress has been made toward the development of the GOLD system, and this paper describes the objectives of this program, basic design of the GOLD system, and results from initial ground-based atmospheric tests.

  15. Fluoride Salt-Cooled High-Temperature Reactor Technology Development and Demonstration Roadmap

    SciTech Connect

    Holcomb, David Eugene; Flanagan, George F; Mays, Gary T; Pointer, William David; Robb, Kevin R; Yoder Jr, Graydon L

    2013-11-01

    Fluoride salt-cooled High-temperature Reactors (FHRs) are an emerging reactor class with potentially advantageous performance characteristics, and fully passive safety. This roadmap describes the principal remaining FHR technology challenges and the development path needed to address the challenges. This roadmap also provides an integrated overview of the current status of the broad set of technologies necessary to design, evaluate, license, construct, operate, and maintain FHRs. First-generation FHRs will not require any technology breakthroughs, but do require significant concept development, system integration, and technology maturation. FHRs are currently entering early phase engineering development. As such, this roadmap is not as technically detailed or specific as would be the case for a more mature reactor class. The higher cost of fuel and coolant, the lack of an approved licensing framework, the lack of qualified, salt-compatible structural materials, and the potential for tritium release into the environment are the most obvious issues that remain to be resolved.

  16. External Events Analysis for LWRS/RISMC Project: Methodology Development and Early Demonstration

    SciTech Connect

    Parisi, Carlo; Prescott, Steven Ralph; Yorg, Richard Alan; Coleman, Justin Leigh; Szilard, Ronaldo Henriques

    2016-02-01

    The ultimate scope of Industrial Application #2 (IA) of the LWRS/RISMC project is a realistic simulation of natural external hazards that impose threat to a NPP. This scope requires the development of a methodology and of a qualified set of tools able to perform advanced risk- informed safety analysis. In particular the methodology should be able to combine results from seismic, flooding and thermal-hydraulic (TH) deterministic calculations with dynamic PRA. This summary presents the key points of methodology being developed and the very first sample application of it to a simple problem (spent fuel pool).

  17. Multi-Lab EV Smart Grid Integration Requirements Study. Providing Guidance on Technology Development and Demonstration

    SciTech Connect

    Markel, T.; Meintz, A.; Hardy, K.; Chen, B.; Bohn, T.; Smart, J.; Scoffield, D.; Hovsapian, R.; Saxena, S.; MacDonald, J.; Kiliccote, S.; Kahl, K.; Pratt, R.

    2015-05-28

    The report begins with a discussion of the current state of the energy and transportation systems, followed by a summary of some VGI scenarios and opportunities. The current efforts to create foundational interface standards are detailed, and the requirements for enabling PEVs as a grid resource are presented. Existing technology demonstrations that include vehicle to grid functions are summarized. The report also includes a data-based discussion on the magnitude and variability of PEVs as a grid resource, followed by an overview of existing simulation tools that vi This report is available at no cost from the National Renewable Energy Laboratory (NREL) at www.nrel.gov/publications. can be used to explore the expansion of VGI to larger grid functions that might offer system and customer value. The document concludes with a summary of the requirements and potential action items that would support greater adoption of VGI.

  18. Development, testing, and demonstration of an optimal fine coal cleaning circuit

    SciTech Connect

    Mishra, M.; Placha, M.; Bethell, P.

    1995-11-01

    The overall objective of this project is to improve the efficiency of fine coal cleaning. The project will be completed in two phases: bench-scale testing and demonstration of four advanced flotation cells and; in-plant proof-of-concept (POC) pilot plant testing of two flotation cells individually and in two-stage combinations. The goal is to ascertain if a two-stage circuit can result in reduced capital and operating costs while achieving improved separation efficiency. The plant selected for this project, Cyprus Emerald Coal Preparation plant, cleans 1200 tph of raw coal. The plant produces approximately 4 million tonnes of clean coal per year at an average as received energy content of 30.2 MJ/Kg (13,000 Btu/lb).

  19. DEVELOPMENT OF UREA-SCR FOR HEAVY-DUTY TRUCKS DEMONSTRATION UPDATE

    SciTech Connect

    Miller, William

    2000-08-20

    This study included engine cell and vehicle tests. The engine cell tests are aimed at determining NOX reduction using the US transient and OICA emissions test cycles. These cycles will be included in future US HD emissions standards. The vehicle tests will show urea-SCR system performance during real-world operation. These tests will prove that the technology can be successfully implemented and demonstrated over-the-road. The program objectives are to: (a) apply urea-SCR to a US HD diesel engine; (b) determine engine cell emissions reduction during US-transient and OICA cycles; (c) apply urea-SCR to a US HD diesel truck; and (d) determine NOX reduction and urea consumption during over-the-road operation.

  20. Development of a demonstration power plant by ocean thermal energy conversion (OTEC)

    SciTech Connect

    Ito, F.; Takazawa, K.; Terayama, T.

    1984-01-01

    At the opening ceremony, the system was praised by leading figures invited from the Oceanic non-oil-producing countries. The power generation test of the OTEC demonstration plant was completed with many new records attained. As engineers who have participated in this project, the authors believe that they have gained confidence in their ability to construct a first-stage commercial OTEC plant of the built-on-land type, though admitting that there still remain some points to be improved. Subjects requiring further study are improvements of material and installation methods enabling the use of water intake piping with larger diameters, further improvement of heat transfer performance at the seawater side (tube inside) of the heat transfer tubes, etc. Since the commercialization of an OTEC system depends mainly on the economical level of the system, cost reduction in the manufacture of equipment and construction is also required.

  1. DEMONSTRATION BULLETIN: AOSTRA-SOILTECH ANAEROBIC THERMAL PROCESSOR: WIDE BEACH DEVELOPMENT SITE

    EPA Science Inventory

    The anaerobic thermal processor (ATP) was developed by UMATAC Industrial Processes under the sponsorship of the Alberta Oil Sands Technology and Research Authority (AOSTRA) and is licensed by SoilTech ATP Systems, Inc., a U.S. corporation. The ATP technology involves a physi...

  2. Nb3Sn RRP® strand and Rutherford cable development for a 15 T dipole demonstrator

    DOE PAGES

    Barzi, E.; Andreev, N.; Li, P.; ...

    2016-03-16

    Keystoned Rutherford cables made of 28 strands and with a stainless steel core were developed and manufactured using 1 mm Nb3Sn composite wires produced by Oxford Superconducting Technology with 127 and 169 restacks using the Restacked-Rod-Process®. Furthermore, the performance and properties of these cables were studied to evaluate possible candidates for 15 T accelerator magnets.

  3. Demonstrating the Benefits of Staff Development: Assessment When Beginning a SIP Process.

    ERIC Educational Resources Information Center

    Todnem, Guy R.; Warner, Michael P.

    1995-01-01

    Assessment can assist key steps in the development of effective school improvement planning (SIP) programs. The article refers to an SIP Action Guide, highlighting four occasions during use of the Action Guide when assessment processes are key (assessment within action plans and assessment to identify strengths, concerns, and possible causes). (SM)

  4. DEVELOPMENT AND DEMONSTRATION OF A BIDIRECTIONAL ADVECTIVE FLUX METER FOR SEDIMENT-WATER INTERFACE

    EPA Science Inventory

    A bidirectional advective flux meter for measuring water transport across the sediment-water interface has been successfully developed and field tested. The flow sensor employs a heat-pulse technique combined with a flow collection funnel for the flow measurement. Because the dir...

  5. Research, development, and demonstration of lead-acid batteries for electric vehicle propulsion. Annual report, 1980

    SciTech Connect

    Not Available

    1981-03-01

    The progress and status of Eltra's Electric Vehicle Battery Program during FY-80 are presented under five divisional headings: Research on Components and Processes; Development of Cells and Modules for Electric Vehicle Propulsion; Sub-Systems; Pilot Line Production of Electric Vehicle Battery Prototypes; and Program Management.

  6. Demonstration Project on Developing Alternative Qualifications and Credentials for Paraprofessionals. Phase II. July 1970-December 1973. Final Report.

    ERIC Educational Resources Information Center

    Lesh, Seymour; And Others

    This demonstration aims at eliminating major obstacles blocking career advancement of human service paraprofessionals. It seeks to develop new options for upgrading workers in four occupations: addiction services, child development, occupational therapy, and teaching. The policies and practices of educational institutions, employers, unions,…

  7. Runway exit designs for capacity improvement demonstrations. Phase 2: Computer model development

    NASA Technical Reports Server (NTRS)

    Trani, A. A.; Hobeika, A. G.; Kim, B. J.; Nunna, V.; Zhong, C.

    1992-01-01

    The development is described of a computer simulation/optimization model to: (1) estimate the optimal locations of existing and proposed runway turnoffs; and (2) estimate the geometric design requirements associated with newly developed high speed turnoffs. The model described, named REDIM 2.0, represents a stand alone application to be used by airport planners, designers, and researchers alike to estimate optimal turnoff locations. The main procedures are described in detail which are implemented in the software package and possible applications are illustrated when using 6 major runway scenarios. The main output of the computer program is the estimation of the weighted average runway occupancy time for a user defined aircraft population. Also, the location and geometric characteristics of each turnoff are provided to the user.

  8. Development and Demonstration of an Innovative Thermal Energy Storage System for Baseload Power Generation

    SciTech Connect

    Goswami, D. Yogi

    2012-09-04

    The objective of this project is to research and develop a thermal energy storage system (operating range 300°C - 450°C) based on encapsulated phase change materials (PCM) that can meet the utility-scale base-load concentrated solar power plant requirements at much lower system costs compared to the existing thermal energy storage (TES) concepts. The major focus of this program is to develop suitable encapsulation methods for existing low-cost phase change materials that would provide a cost effective and reliable solution for thermal energy storage to be integrated in solar thermal power plants. This project proposes a TES system concept that will allow for an increase of the capacity factor of the present CSP technologies to 75% or greater and reduce the cost to less than $20/kWht.

  9. Research, Development and Demonstration of Micro-CHP Systems for Residential Applications - Phase I

    SciTech Connect

    Robert A. Zogg

    2011-03-14

    The objective of the Micro-CHP Phase I effort was to develop a conceptual design for a Micro-CHP system including: Defining market potential; Assessing proposed technology; Developing a proof-of-principle design; and Developing a commercialization strategy. TIAX LLC assembled a team to develop a Micro-CHP system that will provide electricity and heating. TIAX, the contractor and major cost-share provider, provided proven expertise in project management, prime-mover design and development, appliance development and commercialization, analysis of residential energy loads, technology assessment, and market analysis. Kohler Company, the manufacturing partner, is a highly regarded manufacturer of standby power systems and other residential products. Kohler provides a compellingly strong brand, along with the capabilities in product development, design, manufacture, distribution, sales, support, service, and marketing that only a manufacturer of Kohler's status can provide. GAMA, an association of appliance and equipment manufacturers, provided a critical understanding of appliance commercialization issues, including regulatory requirements, large-scale market acceptance issues, and commercialization strategies. The Propane Education & Research Council, a cost-share partner, provided cost share and aided in ensuring the fuel flexibility of the conceptual design. Micro-CHP systems being commercialized in Europe and Japan are generally designed to follow the household thermal load, and generate electricity opportunistically. In many cases, any excess electricity can be sold back to the grid (net metering). These products, however, are unlikely to meet the demands of the U.S. market. First, these products generally cannot provide emergency power when grid power is lost--a critical feature to market success in the U.S. Even those that can may have insufficient electric generation capacities to meet emergency needs for many U.S. homes. Second, the extent to which net metering

  10. X-37 Flight Demonstrator Project: Capabilities for Future Space Transportation System Development

    NASA Technical Reports Server (NTRS)

    Dumbacher, Daniel L.

    2004-01-01

    The X-37 Approach and Landing Vehicle (ALTV) is an automated (unmanned) spacecraft designed to reduce technical risk in the descent and landing phases of flight. ALTV mission requirements and Orbital Vehicle (OV) technology research and development (R&D) goals are formulated to validate and mature high-payoff ground and flight technologies such as Thermal Protection Systems (TPS). It has been more than three decades since the Space Shuttle was designed and built. Real-world hardware experience gained through the multitude of X-37 Project activities has expanded both Government and industry knowledge of the challenges involved in developing new generations of spacecraft that can fulfill the Vision for Space Exploration.

  11. Design, Development, and Demonstration of a Prognostic and Diagnostics Health Monitoring System for the CROWS Platform

    DTIC Science & Technology

    2010-06-01

    assembly, and for giving invaluable feedback on design approaches and options. 1 1. Introduction As the U.S. Army continues towards a...needs to be replaced by a more robust and complete commercial driver, which is very likely to be part of a selected RTOS. Efl -dosfs This directory...effectively gives the user real-time feedback on the state of the desired PDSM boards. Further development should look more carefully at how information is

  12. Cummins Engine Company B5.9 Propane Engine Development, Certification, and Demonstration Project

    SciTech Connect

    The ADEPT Group, Inc.

    1998-12-18

    The objective of this project was to successfuly develop and certify an LPG-dedicated medium-duty original equipment manufacturer (OEM) engine that could be put into production. The engine was launched into production in 1994, and more than 800 B5.9G engines are now in service in the United States and abroad. This engine is now offered by more than 30 bus and truck OEMs.

  13. National Aero-Space Plane: A Technology Development and Demonstration Program to Build the X-30.

    DTIC Science & Technology

    1988-04-01

    8217 s How Is the NASP Program’s Management Strategy 25 Designed to Reduce Technological, Programmatic, Management Structure and Financial Risks ? and...key technological developments, integration, and risks , (4) potential military, space, and commercial mission applications, (5) pro- gram management ...Page 21 GAO NSIAD-, 122 National Ae-Space Plane .t. Chapter 2 What Is the NASP Program’s Management Structure and Its Strategy to Reduce Risks ? The

  14. Research, development and demonstration of lead-acid batteries for electric vehicle propulsion. Annual report, 1979

    SciTech Connect

    Not Available

    1980-06-01

    This report describes work performed from October 1, 1978 to September 30, 1979. The approach for development of both the Improved State-of-the-Art (ISOA) and Advanced lead-acid batteries is three pronged. This approach concentrates on simultaneous optimization of battery design, materials, and manufacturing processing. The 1979 fiscal year saw the achievement of significant progress in the program. Some of the major accomplishments of the year are outlined. 33 figures, 13 tables. (RWR)

  15. Power-by-Wire Development and Demonstration for Subsonic Civil Transport

    NASA Technical Reports Server (NTRS)

    1996-01-01

    During the last decade, three significant studies by the Lockheed Martin Corporation, the NASA Lewis Research Center, and McDonnell Douglas Corporation have clearly shown operational, weight, and cost advantages for commercial subsonic transport aircraft that use all-electric or more-electric technologies in the secondary electric power systems. Even though these studies were completed on different aircraft, used different criteria, and applied a variety of technologies, all three have shown large benefits to the aircraft industry and to the nation's competitive position. The Power-by-Wire (PBW) program is part of the highly reliable Fly-By-Light/Power-By-Wire (FBL/PBW) Technology Program, whose goal is to develop the technology base for confident application of integrated FBL/PBW systems for transport aircraft. This program is part of the NASA aeronautics strategic thrust in subsonic aircraft/national airspace (Thrust 1) to "develop selected high-leverage technologies and explore new means to ensure the competitiveness of U.S. subsonic aircraft and to enhance the safety and productivity of the national aviation system" (The Aeronautics Strategic Plan). Specifically, this program is an initiative under Thrust 1, Key Objective 2, to "develop, in cooperation with U.S. industry, selected high-payoff technologies that can enable significant improvements in aircraft efficiency and cost."

  16. Development and demonstration of direct carbonate fuel cell systems at Energy Research Corporation

    SciTech Connect

    Leo, A.J.; Kush, A.K.; Farooque, M.

    1996-12-31

    Energy Research Corporation (ERC) has been pursuing the development of the direct carbonate fuel cell (DFC) for commercialization near the end of this decade. The DFC produces power directly from hydrocarbon fuels electrochemically, without the need for external reforming or intermediate mechanical conversion steps. As a result, the DFC has the potential to achieve very high efficiency with very low levels of environmental emissions. Modular DFC power plants, which can be shop-fabricated and sited near the user, are ideally suited for distributed generation, industrial, cogeneration, and defense applications. ERC has selected a 2.85 MW power plant unit for initial market entry. Significant advances have been made at ERC in the areas of cell and stack technology and system optimization. Development activities have progressed to the point where 130 kW stacks have been tested in ERC`s subscale power plant, and subscale stacks have been tested in utility and industrial sites around the world. In addition, the world`s first multi-megawatt scale DFC power plant was recently started. Two ERC subsidiaries have been formed to advance the commercialization effort: the Fuel Cell Manufacturing Corporation (FCMC) and the Fuel Cell Engineering Corporation (FCE). FCMC manufacturers carbonate stacks and multi-stack modules, currently from its manufacturing facility in Torrington, CT. FCE is responsible for power plant design, integration of all subsystems, sales/marketing, and client services. This paper describes the results of ERC`s ongoing development and commercialization efforts.

  17. Development, Demonstration, and Analysis of an Integrated Iodine Hall Thruster Feed System

    NASA Technical Reports Server (NTRS)

    Polzin, Kurt A.; Peeples, Steven R.; Burt, Adam O.; Martin, Adam K.; Martinez, Armando; Seixal, Joao F.; Mauro, Stephanie

    2016-01-01

    The design of an in-space iodine-vapor-fed Hall effect thruster propellant management system is described. The solid-iodine propellant tank has unique issues associated with the microgravity environment, requiring a solution where the iodine is maintained in intimate thermal contact with the heated tank walls. The flow control valves required alterations from earlier iterations to survive for extended periods of time in the corrosive iodine-vapor environment. Materials have been selected for the entire feed system that can chemically resist the iodine vapor, with the design now featuring Hastelloy or Inconel for almost all the wetted components. An integrated iodine feed system/Hall thruster demonstration unit was fabricated and tested, with all control being handled by an onboard electronics card specifically designed to operate the feed system. Structural analysis shows that the feed system can survive launch loads after the implementation of some minor reinforcement. Flow modeling, while still requiring significant additional validation, is presented to show its potential in capturing the behavior of components in this low-flow, low-pressure system.

  18. Demonstration of space optical transmitter development for multiple high-frequency bands

    NASA Astrophysics Data System (ADS)

    Nguyen, Hung; Simons, Rainee; Wintucky, Edwin; Freeman, Jon

    2013-05-01

    As the demand for multiple radio frequency carrier bands continues to grow in space communication systems, the design of a cost-effective compact optical transmitter that is capable of transmitting selective multiple RF bands is of great interest, particularly for NASA Space Communications Network Programs. This paper presents experimental results that demonstrate the feasibility of a concept based on an optical wavelength division multiplexing (WDM) technique that enables multiple microwave bands with different modulation formats and bandwidths to be combined and transmitted all in one unit, resulting in many benefits to space communication systems including reduced size, weight and complexity with corresponding savings in cost. Experimental results will be presented including the individual received RF signal power spectra for the L, C, X, Ku, Ka, and Q frequency bands, and measurements of the phase noise associated with each RF frequency. Also to be presented is a swept RF frequency power spectrum showing simultaneous multiple RF frequency bands transmission. The RF frequency bands in this experiment are among those most commonly used in NASA space environment communications.

  19. Demonstration of Space Optical Transmitter Development for Multiple High Frequency Bands

    NASA Technical Reports Server (NTRS)

    Nguyen, Hung; Simons, Rainee; Wintucky, Edwin; Freeman, Jon

    2013-01-01

    As the demand for multiple radio frequency carrier bands continues to grow in space communication systems, the design of a cost-effective compact optical transmitter that is capable of transmitting selective multiple RF bands is of great interest, particularly for NASA Space Communications Network Programs. This paper presents experimental results that demonstrate the feasibility of a concept based on an optical wavelength division multiplexing (WDM) technique that enables multiple microwave bands with different modulation formats and bandwidths to be combined and transmitted all in one unit, resulting in many benefits to space communication systems including reduced size, weight and complexity with corresponding savings in cost. Experimental results will be presented including the individual received RF signal power spectra for the L, C, X, Ku, Ka, and Q frequency bands, and measurements of the phase noise associated with each RF frequency. Also to be presented is a swept RF frequency power spectrum showing simultaneous multiple RF frequency bands transmission. The RF frequency bands in this experiment are among those most commonly used in NASA space environment communications.

  20. Development of derived investigation levels for use in internal dosimetry at the West Valley Demonstration Project

    SciTech Connect

    Johnson, P.

    1991-12-31

    The objective was to determine if the routine intemal dosimetry program at the West Valley Demonstration Project is capable of meeting the performance objective of 1 mSv annual effective dose equivalent due to internal contamination. With the use of the computer code REMedy the annual effective dose equivalent is calculated. Some of the radionuclides of concern result in an annual effective dose equivalent that exceeds the performance objective. Although the results exceed the performance objective, in all but two cases they do not exceed the US DOE regulatory limits. In these instances the Th-232 and Am-241 were determined to exceed the committed dose equivalent limit to their limiting tissue. In order to document the potential missed dose for regulatory compliance, Sr-90 is used as an indicator for Th-232. For Am-241 an investigation as to whether or not the minimum detectable amount can be lowered is performed. The derived investigation levels as a result of this project are 4.9E3 Bq/lung count for Co-60, 2.2E4 Bq/lung count for Cs-137, 1.9 Bq/1 for Sr-90 and for radionuclides other than Sr-90 any value greater than or equal to three standard deviations above their net count is considered to require further investigation.

  1. Enhanced heterologous expression of two Streptomyces griseolus cytochrome P450s and Streptomyces coelicolor ferredoxin reductase as potentially efficient hydroxylation catalysts.

    PubMed

    Hussain, Haitham A; Ward, John M

    2003-01-01

    The herbicide-inducible, soluble cytochrome P450s CYP105A1 and CYP105B1 and their adjacent ferredoxins, Fd1 and Fd2, of Streptomyces griseolus were expressed in Escherichia coli to high levels. Conditions for high-level expression of active enzyme able to catalyze hydroxylation have been developed. Analysis of the expression levels of the P450 proteins in several different E. coli expression hosts identified E. coli BL21 Star(DE3)pLysS as the optimal host cell to express CYP105B1 as judged by CO difference spectra. Examination of the codons used in the CYP1051A1 sequence indicated that it contains a number of codons corresponding to rare E. coli tRNA species. The level of its expression was improved in the modified forms of E. coli BL21(DE3), which contain extra copies of rare codon E. coli tRNA genes. The activity of correctly folded cytochrome P450s was further enhanced by cloning a ferredoxin reductase from Streptomyces coelicolor downstream of CYP105A1 and CYP105B1 and their adjacent ferredoxins. Expression of CYP105A1 and CYP105B1 was also achieved in Streptomyces lividans 1326 by cloning the P450 genes and their ferredoxins into the expression vector pBW160. S. lividans 1326 cells containing CYP105A1 or CYP105B1 were able efficiently to dealkylate 7-ethoxycoumarin.

  2. A novel function of Streptomyces integration host factor (sIHF) in the control of antibiotic production and sporulation in Streptomyces coelicolor.

    PubMed

    Yang, Yung-Hun; Song, Eunjung; Willemse, Joost; Park, Sung-Hee; Kim, Woo-Seong; Kim, Eun-jung; Lee, Bo-Rahm; Kim, Ji-Nu; van Wezel, Gilles P; Kim, Byung-Gee

    2012-03-01

    Bacterial integration host factors (IHFs) play important roles in site-specific recombination, DNA replication, transcription, genome organization and bacterial pathogenesis. In Streptomyces coelicolor, there are three putative IHFs: SCO1480, SCO2950 and SCO5556. SCO1480 or Streptomyces IHF (sIHF) was previously identified as a transcription factor that binds to the promoter region of redD, the pathway-specific regulatory gene for the undecylprodigiosin biosynthetic gene cluster. Here we show that production of the pigmented antibiotics actinorhodin and undecylprodigiosin is strongly enhanced in sihf null mutants, while sporulation was strongly inhibited, with an on average 25% increase in spore size. Furthermore, the sihf mutant spores showed strongly reduced viability, with high sensitivity to heat and live/dead staining revealing a high proportion of empty spores, while enhanced expression of sIHF increased viability. This suggests a major role for sIHF in controlling viability, perhaps via the control of DNA replication and/or segregation. Proteomic analysis of the sihf null mutant identified several differentially expressed transcriptional regulators, indicating that sIHF may have an extensive response regulon. These data surprisingly reveal that a basic architectural element conserved in many actinobacteria such as mycobacteria, corynebacteria, streptomycetes and rhodococci may act as a global regulator of secondary metabolism and cell development.

  3. Engineering of N-acetylglucosamine metabolism for improved antibiotic production in Streptomyces coelicolor A3(2) and an unsuspected role of NagA in glucosamine metabolism

    PubMed Central

    Świątek, Magdalena A.; Urem, Mia; Tenconi, Elodie; Rigali, Sebastien; van Wezel, Gilles P.

    2012-01-01

    N-acetylglucosamine (GlcNAc), the monomer of chitin and constituent of bacterial peptidoglycan, is a preferred carbon and nitrogen source for streptomycetes. Recent studies have revealed new functions of GlcNAc in nutrient signaling of bacteria. Exposure to GlcNAc activates development and antibiotic production of Streptomyces coelicolor under poor growth conditions (famine) and blocks these processes under rich conditions (feast). Glucosamine-6-phosphate (GlcN-6P) is a key molecule in this signaling pathway and acts as an allosteric effector of a pleiotropic transcriptional repressor DasR, the regulon of which includes the GlcNAc metabolic enzymes N-actetylglucosamine-6-phosphate (GlcNAc-6P) deacetylase (NagA) and GlcN-6P deaminase (NagB). Intracellular accumulation of GlcNAc-6P and GlcN-6P enhanced production of the pigmented antibiotic actinorhodin. When the nagB mutant was challenged with GlcNAc or GlcN, spontaneous second-site mutations that relieved the toxicity of the accumulated sugar phosphates were obtained. Surprisingly, deletion of nagA also relieved toxicity of GlcN, indicating novel linkage between the GlcN and GlcNAc utilization pathways. The strongly enhanced antibiotic production observed for many suppressor mutants shows the potential of the modulation of GlcNAc and GlcN metabolism as a metabolic engineering tool toward the improvement of antibiotic productivity or even the discovery of novel compounds. PMID:22892576

  4. AllR Controls the Expression of Streptomyces coelicolor Allantoin Pathway Genes

    PubMed Central

    Navone, Laura; Macagno, Juan Pablo; Licona-Cassani, Cuauhtémoc; Marcellin, Esteban; Nielsen, Lars K.; Gramajo, Hugo

    2015-01-01

    Streptomyces species are native inhabitants of soil, a natural environment where nutrients can be scarce and competition fierce. They have evolved ways to metabolize unusual nutrients, such as purines and its derivatives, which are highly abundant in soil. Catabolism of these uncommon carbon and nitrogen sources needs to be tightly regulated in response to nutrient availability and environmental stimulus. Recently, the allantoin degradation pathway was characterized in Streptomyces coelicolor. However, there are questions that remained unanswered, particularly regarding pathway regulation. Here, using a combination of proteomics and genetic approaches, we identified the negative regulator of the allantoin pathway, AllR. In vitro studies confirmed that AllR binds to the promoter regions of allantoin catabolic genes and determined the AllR DNA binding motif. In addition, effector studies showed that allantoic acid, and glyoxylate, to a lesser extent, inhibit the binding of AllR to the DNA. Inactivation of AllR repressor leads to the constitutive expression of the AllR regulated genes and intriguingly impairs actinorhodin and undecylprodigiosin production. Genetics and proteomics analysis revealed that among all genes from the allantoin pathway that are upregulated in the allR mutant, the hyi gene encoding a hydroxypyruvate isomerase (Hyi) is responsible of the impairment of antibiotic production. PMID:26187964

  5. Complex intra-operonic dynamics mediated by a small RNA in Streptomyces coelicolor.

    PubMed

    Hindra; Moody, Matthew J; Jones, Stephanie E; Elliot, Marie A

    2014-01-01

    Streptomyces are predominantly soil-dwelling bacteria that are best known for their multicellular life cycle and their prodigious metabolic capabilities. They are also renowned for their regulatory capacity and flexibility, with each species encoding >60 sigma factors, a multitude of transcription factors, and an increasing number of small regulatory RNAs. Here, we describe our characterization of a conserved small RNA (sRNA), scr4677. In the model species Streptomyces coelicolor, this sRNA is located in the intergenic region separating SCO4677 (an anti-sigma factor-encoding gene) and SCO4676 (a putative regulatory protein-encoding gene), close to the SCO4676 translation start site in an antisense orientation. There appears to be considerable genetic interplay between these different gene products, with wild type expression of scr4677 requiring function of the anti-sigma factor SCO4677, and scr4677 in turn influencing the abundance of SCO4676-associated transcripts. The scr4677-mediated effects were independent of RNase III (a double stranded RNA-specific nuclease), with RNase III having an unexpectedly positive influence on the level of SCO4676-associated transcripts. We have shown that both SCO4676 and SCO4677 affect the production of the blue-pigmented antibiotic actinorhodin under specific growth conditions, and that this activity appears to be independent of scr4677.

  6. A Eukaryote-like Cardiolipin Synthase Is Present in Streptomyces coelicolor and in Most Actinobacteria*

    PubMed Central

    Sandoval-Calderón, Mario; Geiger, Otto; Guan, Ziqiang; Barona-Gómez, Francisco; Sohlenkamp, Christian

    2009-01-01

    Cardiolipin (CL) is an anionic membrane lipid present in bacteria, plants, and animals, but absent from archaea. It is generally thought that bacteria use an enzyme belonging to the phospholipase D superfamily as cardiolipin synthase (Cls) catalyzing a reversible phosphatidyl group transfer from one phosphatidylglycerol (PG) molecule to another PG to form CL and glycerol. In contrast, in eukaryotes a Cls of the CDP-alcohol phosphatidyltransferase superfamily uses cytidine diphosphate-diacylglycerol (CDP-DAG) as the donor of the phosphatidyl group, which is transferred to a molecule of PG to form CL. Searching the genome of the actinomycete Streptomyces coelicolor A3(2) we identified a gene coding for a putative Cls of the CDP-alcohol phosphatidyltransferase superfamily (Sco1389). Here we show that expression of Sco1389 in a CL-deficient Rhizobium etli mutant restores CL formation. In an in vitro assay Sco1389 condenses CDP-DAG with PG to form CL and therefore catalyzes the same reaction as eukaryotic cardiolipin synthases. This is the first time that a CDP-alcohol phosphatidyltransferase from bacteria is shown to be responsible for CL formation. The broad occurrence of putative orthologues of Sco1389 among the actinobacteria suggests that CL synthesis involving a eukaryotic type Cls is common in actinobacteria. PMID:19439403

  7. Geosmin biosynthesis. Streptomyces coelicolor germacradienol/germacrene D synthase converts farnesyl diphosphate to geosmin.

    PubMed

    Jiang, Jiaoyang; He, Xiaofei; Cane, David E

    2006-06-28

    Geosmin is responsible for the characteristic odor of moist soil. Incubation of recombinant germacradienol synthase, encoded by the SCO6073 (SC9B1.20) gene of the Gram-positive soil bacterium Streptomyces coelicolor, with farnesyl diphosphate (2, FPP) in the presence of Mg2+ gave a mixture of (4S,7R)-germacra-1(10)E,5E-diene-11-ol (3) (74%), (-)-(7S)-germacrene D (4) (10%), geosmin (1) (13%), and a hydrocarbon, tentatively assigned the structure of octalin 5 (3%). Individual incubations of recombinant germacradienol synthase with [1,1-2H2]FPP (2a), (1R)-[1-2H]-FPP (2b), and (1S)-[1-2H]-FPP (2c), as well as with FPP (2) in D2O, and GC-MS analysis of the resulting deuterated products supported a mechanism of geosmin formation involving proton-initiated cyclization and retro-Prins fragmentation of the initially formed germacradienol to give intermediate 5, followed by protonation of 5, 1,2-hydride shift, and capture of water.

  8. Biosynthesis of the earthy odorant geosmin by a bifunctional Streptomyces coelicolor enzyme.

    PubMed

    Jiang, Jiaoyang; He, Xiaofei; Cane, David E

    2007-11-01

    Geosmin is responsible for the characteristic odor of moist soil, as well as off-flavors in drinking water and foodstuffs. Geosmin is generated from farnesyl diphosphate (FPP, 2) by an enzyme that is encoded by the SCO6073 gene in the soil organism Streptomyces coelicolor A32 (ref. 3). We have now shown that the recombinant N-terminal half of this protein catalyzes the Mg2+-dependent cyclization of FPP to germacradienol and germacrene D, while the highly homologous C-terminal domain, previously thought to be catalytically silent, catalyzes the Mg2+-dependent conversion of germacradienol to geosmin. Site-directed mutagenesis confirmed that the N- and C-terminal domains each harbor a distinct, independently functioning active site. A mutation in the N-terminal domain of germacradienol-geosmin synthase of a catalytically essential serine to alanine results in the conversion of FPP to a mixture of sesquiterpenes that includes an aberrant product identified as isolepidozene, which was previously suggested to be an enzyme-bound intermediate in the cyclization of FPP to germacradienol.

  9. Development and Demonstration of Smart grid inverters for High-Penetration PV Applications

    SciTech Connect

    Roose, Leon; Liang, Nathan; Noma, Edwin; Tran, Thai; Matsuura, Marc

    2014-10-31

    The objective of this Smart Grid Inverter (SGI) project is to implement, on operating utility distribution feeders with “very high” penetration of rooftop PV, enhanced capability smart inverters to achieve improved operational performance, control and visibility. This is accomplished by creating, deploying, and evaluating new smart inverters using integrated inverter management control software (IMCS) and standards-based communications systems. Detailed distribution modeling is also employed to aid in development of inverter control algorithms/settings. The project will test various inverter control strategies in two project deployment locations – Maui, Hawai’i and Maryland/Washington D.C.

  10. DEVELOPMENT, TESTING, AND DEMONSTRATION OF AN OPTIMAL FINE COAL CLEANING CIRCUIT

    SciTech Connect

    Steven R. Hadley; R. Mike Mishra; Michael Placha

    1999-01-27

    The objective of this project was to improve the efficiency of the fine coal froth flotation circuit in commercial coal preparation plants. The plant selected for this project, Cyprus Emerald Coal Preparation Plant, cleans 1200-1400 tph of Pittsburgh seam raw coal and uses conventional flotation cells to clean the minus 100-mesh size fraction. The amount of coal in this size fraction is approximately 80 tph with an average ash content of 35%. The project was carried out in two phases. In Phase I, four advanced flotation cells, i.e., a Jameson cell, an Outokumpu HG tank cell, an open column, and a packed column cell, were subjected to bench-scale testing and demonstration. In Phase II, two of these flotation cells, the Jameson cell and the packed column, were subjected to in-plant, proof-of-concept (POC) pilot plant testing both individually and in two-stage combination in order to ascertain whether a two-stage circuit results in lower levelized production costs. The bench-scale results indicated that the Jameson cell and packed column cell would be amenable to the single- and two-stage flotation approach. POC tests using these cells determined that single-stage coal matter recovery (CMR) of 85% was possible with a product ash content of 5.5-7%. Two-stage operation resulted in a coal recovery of 90% with a clean coal ash content of 6-7.5%. This compares favorably with the plant flotation circuit recovery of 80% at a clean coal ash of 11%.

  11. Research, development, and demonstration of nickel-iron batteries for electric vehicle propulsion. Annual report, 1979

    SciTech Connect

    Not Available

    1980-06-01

    The program has progressed to the stage of evaluating full-sized (220 Ah) cells, multicell modules, and 22 kWh batteries. Nickel electrodes that display stable capacities of up to 24 Ah/plate (at C/3 drain rate) at design thickness (2.5 mm) in tests at 200/sup +/ test cycles. Iron electrodes of the composite-type are also delivering 24 Ah/plate (at C/3) at target thickness (1.0 mm). Iron plates are displaying capacity stability for 300/sup +/ test cycles in continuing 3 plate cell tests. Best finished cells are delivering 57 to 63 Wh/kg at C/3, based on cell weights of the finished cells, and in the actual designed cell volume. 6-cell module (6-1) performance has demonstrated 239 Ah, 1735 Wh, 53 WH/kg at the C/3 drain rate. This module is now being evaluated at the National Battery Test Laboratory. The 2 x 4 battery has been constructed, tested, and delivered for engineering test and evaluation. The battery delivered 22.5 kWh, as required (199 Ah discharge at 113 V-bar) at the C/3 drain rate. The battery has performed satisfactorily under dynamometer and constant current drain tests. Some cell problems, related to construction, necessitated changing 3 modules, but the battery is now ready for further testing. Reduction in nickel plate swelling (and concurrent stack electrolyte starvation), to improve cycling, is one area of major effort to reach the final battery objectives. Pasted nickel electrodes are showing promise in initial full-size cell tests and will continue to be evaluated in finished cells, along with other technology advancements. 30 figures, 14 tables.

  12. A Cloud-Based Global Flood Disaster Community Cyber-Infrastructure: Development and Demonstration

    NASA Technical Reports Server (NTRS)

    Wan, Zhanming; Hong, Yang; Khan, Sadiq; Gourley, Jonathan; Flamig, Zachary; Kirschbaum, Dalia; Tang, Guoqiang

    2014-01-01

    Flood disasters have significant impacts on the development of communities globally. This study describes a public cloud-based flood cyber-infrastructure (CyberFlood) that collects, organizes, visualizes, and manages several global flood databases for authorities and the public in real-time, providing location-based eventful visualization as well as statistical analysis and graphing capabilities. In order to expand and update the existing flood inventory, a crowdsourcing data collection methodology is employed for the public with smartphones or Internet to report new flood events, which is also intended to engage citizen-scientists so that they may become motivated and educated about the latest developments in satellite remote sensing and hydrologic modeling technologies. Our shared vision is to better serve the global water community with comprehensive flood information, aided by the state-of-the- art cloud computing and crowdsourcing technology. The CyberFlood presents an opportunity to eventually modernize the existing paradigm used to collect, manage, analyze, and visualize water-related disasters.

  13. Combined Heat and Power Systems Technology Development and Demonstration 370 kW High Efficiency Microturbine

    SciTech Connect

    none,

    2015-10-14

    The C370 Program was awarded in October 2010 with the ambitious goal of designing and testing the most electrically efficient recuperated microturbine engine at a rated power of less than 500 kW. The aggressive targets for electrical efficiency, emission regulatory compliance, and the estimated price point make the system state-of-the-art for microturbine engine systems. These goals will be met by designing a two stage microturbine engine identified as the low pressure spool and high pressure spool that are based on derivative hardware of Capstone’s current commercially available engines. The development and testing of the engine occurred in two phases. Phase I focused on developing a higher power and more efficient engine, that would become the low pressure spool which is based on Capstone’s C200 (200kW) engine architecture. Phase II integrated the low pressure spool created in Phase I with the high pressure spool, which is based on Capstone’s C65 (65 kW) commercially available engine. Integration of the engines, based on preliminary research, would allow the dual spool engine to provide electrical power in excess of 370 kW, with electrical efficiency approaching 42%. If both of these targets were met coupled with the overall CHP target of 85% total combined heating and electrical efficiency California Air Resources Board (CARB) level emissions, and a price target of $600 per kW, the system would represent a step change in the currently available commercial generation technology. Phase I of the C370 program required the development of the C370 low pressure spool. The goal was to increase the C200 engine power by a minimum of 25% — 250 kW — and efficiency from 32% to 37%. These increases in the C200 engine output were imperative to meet the power requirements of the engine when both spools were integrated. An additional benefit of designing and testing the C370 low pressure spool was the possibility of developing a stand-alone product for possible

  14. Development and demonstration of a personal monitoring system for exposure to hydrogen fluoride. Final report

    SciTech Connect

    Young, M.S.; Monat, J.P.

    1993-09-01

    A good, functional Hydrogen Fluoride Gasbadge dosimeter has been developed for sampling of airborne HF vapor. The device is small (7.7 cm {times} 5.4 cm {times} 1.9 cm) and can easily and conveniently be worn on one`s lapel. It consists of polyethylene and polypropylene parts and a triethanolamine-impregnated polyproylene collection element. It is completely self contained, requiring no pumps, impingers, or sampling tubes. Subsequent to sampling, the collection element is analyzed quickly and easily with a fluoride selective-ion electrode. Laboratory tests were conducted to determine precision, linearity, interference effects, influences of temperature and humidity, and collection element stability over time. Results of the tests indicate that the Abcor Gasbadge HF dosimeter is an excellent passive HF monitor for work spaces, and that results obtained with it are accurate within {plus_minus}25%. These results have been corroborated in a field study.

  15. Development and Demonstration of a Low Cost Hybrid Drive Train for Medium and Heavy Duty Vehicles

    SciTech Connect

    Strangas, Elias; Schock, Harold; Zhu, Guoming; Moran, Kevin; Ruckle, Trevor; Foster, Shanelle; Cintron-Rivera, Jorge; Tariq, Abdul; Nino-Baron, Carlos

    2011-04-30

    The DOE sponsored effort is part of a larger effort to quantify the efficiency of hybrid powertrain systems through testing and modeling. The focus of the DOE sponsored activity was the design, development and testing of hardware to evaluate the efficiency of the electrical motors relevant to medium duty vehicles. Medium duty hybrid powertrain motors and generators were designed, fabricated, setup and tested. The motors were a permanent magnet configuration, constructed at Electric Apparatus Corporation in Howell, Michigan. The purpose of this was to identify the potential gains in terms of fuel cost savings that could be realized by implementation of such a configuration. As the electric motors constructed were prototype designs, the scope of the project did not include calculation of the costs of mass production of the subject electrical motors or generator.

  16. Implementation of a professional portfolio: a tool to demonstrate professional development for advanced practice.

    PubMed

    Chamblee, Tracy B; Dale, Juanita Conkin; Drews, Barbie; Spahis, Joanna; Hardin, Teri

    2015-01-01

    The literature has a gap related to professional development for APRNs. In the United States, many health care organizations use clinical advancement programs for registered nurses, but APRNs are not often included in these programs. If APRNs are included, advancement opportunities are very limited. At CMC, implementation of a professional portfolio resulted in increased satisfaction among APPs regarding their ability to showcase professional growth and expertise, as well as the uniqueness of their advanced practice. Use of the professional portfolio led to improved recognition by APS and organizational leaders of APP performance excellence during the annual performance evaluation, as well as improved recognition among APP colleagues in terms of nominations for honors and awards.

  17. Research, development, and demonstration of lead-acid batteries for electric vehicle propulsion. Annual report, 1979

    SciTech Connect

    Not Available

    1980-06-01

    The initial phase of work comprises three factorial experiments to evaluate a variety of component combinations. Goals to be met by these batteries include the following: capacity at 3 h discharge, 20 to 30 kWh; specific energy, 40 Wh/kg; specific power, 1000 W/kg for 15 s; cycle life, 800 cycles to 80% depth; price, $50/kWh. The status of the factorial experiments is reviewed. The second phase of work, design of an advanced battery, has the following goals: 30 to 40 kWh; 60 Wh/kg; 150 W/kg for 15 s; 1000 cycles to 80% depth; $40/kWh. It is not yet possible to say whether these goals can be met. Numerous approaches are under study to increase the utilization of battery chemicals. A battery design with no live electrical connection above the battery is being developed. 52 figures, 52 tables. (RWR)

  18. Development and Demonstration of a Magnesium-Intensive Vehicle Front-End Substructure

    SciTech Connect

    Logan, Stephen D.; Forsmark, Joy H.; Osborne, Richard

    2016-07-01

    This project is the final phase (designated Phase III) of an extensive, nine-year effort with the objectives of developing a knowledge base and enabling technologies for the design, fabrication and performance evaluation of magnesium-intensive automotive front-end substructures intended to partially or completely replace all-steel comparators, providing a weight savings approaching 50% of the baseline. Benefits of extensive vehicle weight reduction in terms of fuel economy increase, extended vehicle range, vehicle performance and commensurate reductions in greenhouse gas emissions are well known. An exemplary vehicle substructure considered by the project is illustrated in Figure 1, along with the exterior vehicle appearance. This unibody front-end “substructure” is one physical objective of the ultimate design and engineering aspects established at the outset of the larger collective effort.

  19. Development study of precooled-cycle hypersonic turbojet engine for flight demonstration

    NASA Astrophysics Data System (ADS)

    Sato, Tetsuya; Taguchi, Hideyuki; Kobayashi, Hiroaki; Kojima, Takayuki; Okai, Keiichi; Fujita, Kazuhisa; Masaki, Daisaku; Hongo, Motoyuki; Ohta, Toyohiko

    2007-06-01

    This paper describes a development study of a precooled-cycle hypersonic turbojet engine for the first stage of TSTO space plane and hypersonic airplane. With reflecting the key technologies accumulated from ATREX (expander cycle ATR engine) ground tests, the next flyable subscale engine "S-engine" is now developed. S-engine has 23cm×23cm of rectangular cross-section, 2.2 m of the overall length and about 100 kg of the weight employing a variable-geometry rectangular inlet and nozzle. It produces 1.2 kN of thrust at SLS, which corresponds to {1}/{4} of the ATREX engine. Design of the hypersonic components such as the inlet, precooler and nozzle has been finished and their aerodynamic performances were verified by wind tunnel tests and CFD analyses. A prototype model of the diagonal-flow compressor whose pressure ratio is 6 was manufactured. Its rotating tests under the very-low pressure conditions are now in progress. The reverse-flow annular combustion chamber was successfully tested. The first flight test of the S-engine is to be conducted in 2008 by the balloon-based operation vehicle (BOV) which is about 5 m in length, 0.55 m in diameter and 500 kg in weight. The vehicle is dropped from an altitude of 40 km by a high altitude balloon. After 40-s free-fall, the vehicle pulls up and S-engine operates for 30 s at about Mach 2. High altitude tests of the engine components corresponding to the BOV's flight condition have been conducted.

  20. Evaluation of the professional process portfolio: an innovative tool to help develop and demonstrate leadership competency.

    PubMed

    Hastings, Elisabeth S; Chacko, Mariam R; Acosta, Amy B; Hergenroeder, Albert C; Wiemann, Constance M

    2015-02-01

    The professional process portfolio (PPP) was adopted by the Maternal Child and Health Bureau (MCHB) as an 'innovation' in best practice for all Leadership in Education and Adolescent Health (LEAH) Training Programs; however it had not been formally evaluated. Thus the objective was to evaluate the utility of the PPP for graduates of the LEAH training program in terms of (1) how alumni have used, adapted, and applied it since completing fellowship, (2) what fellows learned or gained through completing it, and (3) how it can be improved for continued use in training programs. Graduates from six disciplines were asked via telephone or email to participate in a survey regarding their experience with the PPP. Descriptive statistics were generated for demographic characteristics and closed-choice questions. Responses to open-ended questions were analyzed by a team of faculty using framework analysis. Sixty-one graduates completed surveys. The majority (85%) found the PPP useful and utilized it post-graduation for multiple purposes in professional development: interviewing, training, and referencing previous work. Graduates recommended that the PPP be improved by making it electronic, discipline-specific, and providing earlier and more frequent instruction from faculty on expectations of creating it. Four themes emerged from the qualitative data analysis: accomplishment, experiential learning, skills and accountability, and a best practice of learning. The PPP was an effective personal learning tool for the majority of graduates and enhanced graduates' experiences. We highlight the ways that the PPP may facilitate the development of learning experiences associated with MCH leadership competence.

  1. Propulsion System Development for the Iodine Satellite (iSAT) Demonstration Mission

    NASA Technical Reports Server (NTRS)

    Polzin, Kurt A.; Peeples, Stephen R.; Seixal, Joao F.; Mauro, Stephanie L.; Lewis, Brandon L.; Jerman, Gregory A.; Calvert, Derek H.; Dankanich, John; Kamhawi, Hani; Hickman, Tyler A.; Szabo, James; Pote, Bruce; Lee, Lauren

    2015-01-01

    The development and testing of a 200-W iodine-fed Hall thruster propulsion system that will be flown on a 12-U CubeSat is described. The switch in propellant from more traditional xenon gas to solid iodine yields the advantage of high density, low pressure propellant storage but introduces new requirements that must be addressed in the design and operation of the propulsion system. The thruster materials have been modified from a previously-flown xenon Hall thruster to make it compatible with iodine vapor. The cathode incorporated into this design additionally requires little or no heating to initiate the discharge, reducing the power needed to start the thruster. The feed system produces iodine vapor in the propellant reservoir through sublimation and then controls the flow to the anode and cathode of the thruster using a pair of proportional flow control valves. The propellant feeding process is controlled by the power processing unit, with feedback control on the anode flow rate provided through a measure of the thruster discharge current. Thermal modeling indicates that it may be difficult to sufficiently heat the iodine if it loses contact with the propellant reservoir walls, serving to motivate future testing of that scenario to verify the modeling result and develop potential mitigation strategies. Preliminary, short-duration materials testing has thus-far indicated that several materials may be acceptable for prolonged contact with iodine vapor, motivating longer-duration testing. A propellant loading procedure is presented that aims to minimize the contaminants in the feed system and propellant reservoir. Finally, an 80-hour duration test being performed to gain experience operating the thruster over long durations and multiple restarts is discussed.

  2. Research development and demonstration of a fuel cell/battery powered bus system

    NASA Astrophysics Data System (ADS)

    Romano, S.; Wimmer, R.

    1993-02-01

    During 1992, the design of the three test bed buses evolved. Georgetown participated in all the working meetings and the preliminary design review of the Test Bed Bus (TBB). Throughout the Year, Georgetown provided input on system design and integration issues, transit industry requirements, and bus performance. GU also attended technical seminars, presenting papers at two. The computer simulation 'HYBRID', developed by Georgetown, was used to assess performance of different bus designs and system control algorithms. GU also modified the simulation to be more flexible and allow easier evaluation of designs. Georgetown had Exhibitgroup design and construct, to our specifications, a fuel cell bus display with a 1/10 scale model of the TBB for use at trade shows and exhibits. Energy Partners of West Palm Beach, Florida completed their subcontract for the engineering design study of a hydrogen fueled, fuel-cell-powered multi-passenger vehicle. Georgetown performed a study to determine the size of the US bus fleet and expected new bus deliveries over the next 10 years. Trojan Battery Company of Santa Fe Springs, California conducted a study to determine the specific design characteristics required of a lead acid battery to meet the special requirements posed by hybrid vehicle operation. Finally, GU assisted the prime contractor with the design of the system controller and its control algorithms.

  3. Development and demonstration of an on-board mission planner for helicopters

    NASA Technical Reports Server (NTRS)

    Deutsch, Owen L.; Desai, Mukund

    1988-01-01

    Mission management tasks can be distributed within a planning hierarchy, where each level of the hierarchy addresses a scope of action, and associated time scale or planning horizon, and requirements for plan generation response time. The current work is focused on the far-field planning subproblem, with a scope and planning horizon encompassing the entire mission and with a response time required to be about two minutes. The far-feld planning problem is posed as a constrained optimization problem and algorithms and structural organizations are proposed for the solution. Algorithms are implemented in a developmental environment, and performance is assessed with respect to optimality and feasibility for the intended application and in comparison with alternative algorithms. This is done for the three major components of far-field planning: goal planning, waypoint path planning, and timeline management. It appears feasible to meet performance requirements on a 10 Mips flyable processor (dedicated to far-field planning) using a heuristically-guided simulated annealing technique for the goal planner, a modified A* search for the waypoint path planner, and a speed scheduling technique developed for this project.

  4. Runway Exit Designs for Capacity Improvement Demonstrations. Phase 1: Algorithm Development

    NASA Technical Reports Server (NTRS)

    Trani, A. A.; Hobeika, A. G.; Sherali, H.; Kim, B. J.; Sadam, C. K.

    1990-01-01

    A description and results are presented of a study to locate and design rapid runway exits under realistic airport conditions. The study developed a PC-based computer simulation-optimization program called REDIM (runway exit design interactive model) to help future airport designers and planners to locate optimal exits under various airport conditions. The model addresses three sets of problems typically arising during runway exit design evaluations. These are the evaluations of existing runway configurations, addition of new rapid runway turnoffs, and the design of new runway facilities. The model is highly interactive and allows a quick estimation of the expected value of runway occupancy time. Aircraft populations and airport environmental conditions are among the multiple inputs to the model to execute a viable runway location and geometric design solution. The results presented suggest that possible reductions on runway occupancy time (ROT) can be achieved with the use of optimally tailored rapid runway designs for a given aircraft population. Reductions of up to 9 to 6 seconds are possible with the implementation of 30 m/sec variable geometry exits.

  5. Pilot and Repeat Trials as Development Tools Associated with Demonstration of Bioequivalence.

    PubMed

    Fuglsang, Anders

    2015-05-01

    The purpose of this work is to use simulated trials to study how pilot trials can be implemented in relation to bioequivalence testing, and how the use of the information obtained at the pilot stage can influence the overall chance of showing bioequivalence (power) or the chance of approving a truly bioinequivalent product (type I error). The work also covers the use of repeat pivotal trials since the difference between a pilot trial followed by a pivotal trial and a pivotal trial followed by a repeat trial is mainly a question of whether a conclusion of bioequivalence can be allowed after the first trial. Repeating a pivotal trial after a failed trial involves dual or serial testing of the bioequivalence null hypothesis, and the paper illustrates how this may inflate the type I error up to almost 10%. Hence, it is questioned if such practice is in the interest of patients. Tables for power, type I error, and sample sizes are provided for a total of six different decision trees which allow the developer to use either the observed geometric mean ratio (GMR) from the first or trial or to assume that the GMR is 0.95. In cases when the true GMR can be controlled so as not to deviate more from unity than 0.95, sequential design methods ad modum Potvin may be superior to pilot trials. The tables provide a quantitative basis for choosing between sequential designs and pivotal trials preceded by pilot trials.

  6. Variable Cycle Engine Control System Definition Study. Turbine Engine Technology Demonstrator Component Development Program, Project 668A. Controls Development Program

    DTIC Science & Technology

    1976-03-01

    JULY 1973 to MARCH 1976 Approved for public rtlease; distribution unlimited DDC R 191r Alit V0tCE, AKt *IRO-10ROU14,40N LAHIORATf0lty .71 L y ~j Alit...engine parameters to "scheduled" optimal poramelric relationships successfully attained minimum sfc and maximum thrust at all the selected test points in... test plan woa formulated for the JTD control system. Table of Contents Page 1.0 Introduction 1-1 2.0 Summary 2- 1 3.0 Joint Technology Demonstrator

  7. Program mid-year summaries research, development, demonstration, testing and evaluation: Office of Technology Development, FY 1993

    SciTech Connect

    Not Available

    1993-10-01

    This mid-year review provides a summary of activities within the Office of Technology Development with individual presentations being made to DOE HQ and field management staff. The presentations are by EM-541, 542, 551, and 552 organizations.

  8. Development and demonstration of an enhanced spreadsheet-based well log analysis software. Final report, May 1998

    SciTech Connect

    Watney, W.L.; Doveton, J.H.; Guy, W.J.

    1998-10-01

    The Advanced Class Work Program is a field-based research and demonstration program for demonstration of advanced or improved technologies identified in the Department of Energy`s Class Field Demonstration Projects. The objective of the Advanced Class Work program is to conduct field demonstrations of technologies for which a small, incremental amount of work will produce or improve a transferable, useful technology for oil recovery. The goal of the program is to enhance the products of near-term Class projects and maximize the applicability and effectiveness of project results. PfEFFER (Petrofacies Evaluation of Formations For Engineering Reservoirs) is a well log analysis computer package. The software was tested and successfully applied in Schaben Field, a DOE Class 2 Field Demonstration Project to assist in improving reservoir characterization and assessing reservoir performance. PfEFFER v.1 was released in January, 1996 as a commercial spreadsheet-based well-log analysis program developed and distributed through the Kansas Geological Survey. The objectives of this project were: Task 1 -- Enhance the PfEFFER software package; Task 2 -- Develop major new modules to significantly augment PfEFFER capabilities; Task 3 -- Conduct field demonstration of software application using the necessary reservoir data acquired from oil operators and construct a database; and Task 4 -- Perform technology transfer activities that include workshops, reports, presentations, or other methods to communicate results to interested parties.

  9. Isolation and expression of the catA gene encoding the major vegetative catalase in Streptomyces coelicolor Müller.

    PubMed Central

    Cho, Y H; Roe, J H

    1997-01-01

    We isolated the catA gene for the major vegetative catalase from Streptomyces coelicolor Müller. It encodes a polypeptide of 488 residues (55,440 Da) that is highly homologous to typical monofunctional catalases. We investigated catA expression by analyzing both catA mRNA and catalase activity. catA expression was increased by H2O2 treatment but did not increase during stationary phase. A putative catalase (CatB) cross-reactive with anti-CatA antibody appeared during stationary phase and in the aerial mycelium. PMID:9190825

  10. GLASS FORMULATION DEVELOPMENT AND TESTING FOR COLD CRUCIBLE INDUCTION MELTER (CCIM) ADVANCED REMEDIATION TECHNOLOGIES DEMONSTRATION PROJECT - 9208

    SciTech Connect

    Marra, J; Amanda Billings, A; David Peeler, D; Michael Stone, M; Tommy Edwards, T

    2008-08-27

    Over the past few years, Cold Crucible Induction Melter (CCIM) demonstrations have been completed using SRS sludge batches 2, 3 and 4 (SB2, SB3 and SB4) simulant compositions. These campaigns demonstrated the ability of the CCIM to effectively produce quality glasses at high waste loadings. The current Advanced Remediation Technology (ART) Phase II-A Project is aimed at demonstrating the CCIM technology under representative DWPF flowsheet conditions and to demonstrate extended operations of the melter. A glass composition development effort was completed to identify and recommend a frit composition and sludge batch 4 (SB4) simulant waste loading target for subsequent ART-Phase II-A CCIM demonstration testing. Based on the results of the glass formulation testing, it was recommended that the Frit 503-R6 composition (B{sub 2}O{sub 3} = 14 wt %; Li{sub 2}O = 9 wt %; Na{sub 2}O = 3 wt %; and SiO{sub 2} = 74 wt %) be utilized for the demonstration. Furthermore, a waste loading of 46 wt % was recommended. The recommended frit and waste loading would produce a glass with acceptable durability with a liquidus temperature adequately below the 1250 C nominal CCIM operating temperature. This frit composition and waste loading was found to result in a glass that met CCIM processing requirements for viscosity, electrical conductivity and thermal conductivity. The recommended frit and waste loading level should also provide a buffer for sludge product compositional variation to support the Phase II-A CCIM demonstration.

  11. Biological restoration of major transportation facilities domestic demonstration and application project (DDAP): technology development at Sandia National Laboratories.

    SciTech Connect

    Ramsey, James L., Jr.; Melton, Brad; Finley, Patrick; Brockman, John; Peyton, Chad E.; Tucker, Mark David; Einfeld, Wayne; Griffith, Richard O.; Brown, Gary Stephen; Lucero, Daniel A.; Betty, Rita G.; McKenna, Sean Andrew; Knowlton, Robert G.; Ho, Pauline

    2006-06-01

    The Bio-Restoration of Major Transportation Facilities Domestic Demonstration and Application Program (DDAP) is a designed to accelerate the restoration of transportation nodes following an attack with a biological warfare agent. This report documents the technology development work done at SNL for this DDAP, which include development of the BROOM tool, an investigation of surface sample collection efficiency, and a flow cytometry study of chlorine dioxide effects on Bacillus anthracis spore viability.

  12. Development and demonstration of manufacturing processes for fabricating graphite/LARC-160 polyimide structural elements, part 4, paragraph B

    NASA Technical Reports Server (NTRS)

    1980-01-01

    A quality assurance program was developed which included specifications for celion/LARC-160 polyimide materials and quality control of materials and processes. The effects of monomers and/or polymer variables and prepeg variables on the processibility of celion/LARC prepeg were included. Processes for fabricating laminates, honeycomb core panels, and chopped fiber moldings were developed. Specimens and conduct tests were fabricated to qualify the processes for fabrication of demonstration components.

  13. Development and field demonstration of an eight-element receive wavelength-multiplexed true-time-delay steering system.

    PubMed

    Goutzoulis, A P; Zomp, J M

    1997-10-10

    We describe the design and development of an eight-element hardware-compressive receive true-time-delay steering system that employs wavelength-division multiplexing. The laboratory system performance and results from the system demonstration at the antenna range are discussed.

  14. 76 FR 35214 - Notice of Determination of Adequacy of Ohio's Research, Development and Demonstration (RD&D...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-16

    ...) Permit Provisions for Municipal Solid Waste Landfills AGENCY: Environmental Protection Agency (EPA..., development and demonstration (RD&D) permits to be issued to certain municipal solid waste landfills by... issued a final rule amending the municipal solid waste landfill criteria in 40 CFR Part 258 to allow...

  15. Light and Plants. A Series of Experiments Demonstrating Light Effects on Seed Germination, Plant Growth, and Plant Development.

    ERIC Educational Resources Information Center

    Downs, R. J.; And Others

    A brief summary of the effects of light on plant germination, growth and development, including photoperiodism and pigment formation, introduces 18 experiments and demonstrations which illustrate aspects of these effects. Detailed procedures for each exercise are given, the expected results outlined, and possible sources of difficulty discussed.…

  16. Development and Demonstration of Mobile, Small Footprint Exploration and Development Well System for Arctic Unconventional Gas Resources (ARCGAS)

    SciTech Connect

    Paul Glavinovich

    2002-11-01

    Traditionally, oil and gas field technology development in Alaska has focused on the high-cost, high-productivity oil and gas fields of the North Slope and Cook Inlet, with little or no attention given to Alaska's numerous shallow, unconventional gas reservoirs (carbonaceous shales, coalbeds, tight gas sands). This is because the high costs associated with utilizing the existing conventional oil and gas infrastructure, combined with the typical remoteness and environmental sensitivity of many of Alaska's unconventional gas plays, renders the cost of exploring for and producing unconventional gas resources prohibitive. To address these operational challenges and promote the development of Alaska's large unconventional gas resource base, new low-cost methods of obtaining critical reservoir parameters prior to drilling and completing more costly production wells are required. Encouragingly, low-cost coring, logging, and in-situ testing technologies have already been developed by the hard rock mining industry in Alaska and worldwide, where an extensive service industry employs highly portable diamond-drilling rigs. From 1998 to 2000, Teck Cominco Alaska employed some of these technologies at their Red Dog Mine site in an effort to quantify a large unconventional gas resource in the vicinity of the mine. However, some of the methods employed were not fully developed and required additional refinement in order to be used in a cost effective manner for rural arctic exploration. In an effort to offset the high cost of developing a new, low-cost exploration methods, the US Department of Energy, National Petroleum Technology Office (DOE-NPTO), partnered with the Nana Regional Corporation and Teck Cominco on a technology development program beginning in 2001. Under this DOE-NPTO project, a team comprised of the NANA Regional Corporation (NANA), Teck Cominco Alaska and Advanced Resources International, Inc. (ARI) have been able to adapt drilling technology developed for the

  17. Development and demonstration of a Stirling/Rankine heat activated heat pump. Phase 3B: Engine technology development testing

    NASA Astrophysics Data System (ADS)

    1984-11-01

    The results of the Phase 3B Stirling/Rankine Heat Activated Heat Pump product development program are given. Results of the Phase 2 program indicated deficiencies in the performance of the free-piston Stirling engine and mismatching of the dynamic characteristics of the engine and the compressor. These deficiencies were further investigated during in-depth diagnostic testing of the engine/compressor unit in the Phase 3B and indicated appropriate engine/compressor matching criteria.

  18. Promoter determining the timing and spatial localization of transcription of a cloned Streptomyces coelicolor gene encoding a spore-associated polypeptide.

    PubMed Central

    Guijarro, J; Santamaria, R; Schauer, A; Losick, R

    1988-01-01

    Streptomyces coelicolor is a filamentous, gram-positive bacterium that exhibits a complex cycle of morphological differentiation involving the formation of an aerial mycelium of multinucleoid hyphae which undergo septation to form long chains of spores. We report the identification of two proteins of 13 and 3 kilodaltons, designated SapA and SapB, respectively, that are produced during formation of the aerial mycelium and are found in assocation with purified, mature spores. We cloned the structural gene (sapA) for one of these spore-associated proteins. Nucleotide sequence analysis suggests that the 13-kilodalton polypeptide is derived from a larger pre- or preproprotein containing a leader sequence of 37 amino acids. Nuclease protection-hybridization analysis and experiments using the Vibrio harveyi, luciferase-encoding luxAB operon as a gene tag demonstrated that expression of sapA is controlled from a promoter contained within a region of less than 110 base pairs in length, whose transcription start site is located approximately 50 base pairs upstream from the initiation codon for the sapA open reading frame. Transcription of sapA was induced at the time of appearance of the aerial mycelium, and the level of sapA transcripts was significantly reduced in certain mutants blocked in aerial mycelium (bld) and or spore (whi) formation. As further evidence of the association of sapA transcription with morphological differentiation, experiments in which we monitored sapA transcription topographically by use of a sapA-luxAB operon fusion demonstrated a close spatial correlation between colony regions undergoing aerial mycelium formation and zones of sapA-promoted light emission. Images PMID:2450872

  19. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1979-01-01

    Presents two demonstrations which are intended for chemistry college students. These demonstrations are: (1) enhancement of concentration quenching by micelles; and (2) the thermite lecture demonstration. (HM)

  20. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1984-01-01

    Procedures for two demonstrations are presented. The first is a demonstration of chemiluminescence. The second is a demonstration using a secondary battery constructed from common household articles. (JN)

  1. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1978-01-01

    Presents the following chemistry lecture demonstrations and experiments: (1) a versatile kinetic demonstration; (2) the Bakelite Demonstration; (3) applying Beer's law; and (4) entropy calculations. (HM)

  2. Development and demonstration of manufacturing processes for fabricating graphite/PMR-15 polyimide structural elements. [space shuttle aft body flap

    NASA Technical Reports Server (NTRS)

    Sheppard, C. H.; Hoggatt, J. T.; Symonds, W. A.

    1979-01-01

    The processing requirements for graphite/PMR-15 polyimide composites developed to demonstrate the structural integrity of polyimide composite structural elements at temperatures up to 589K (600 F) are described. Major tasks included: quality assurance development; materials and process development; specification verification; flat panel fabrication; stiffened panel fabrication; honeycomb panel fabrication; chopped fiber moldings; and demonstration component fabrication. Materials, processing, and quality assurance documents were prepared from experimentally derived data. Structural elements consisting of flat panels, corrugated stiffeners, I-beams, hat stiffeners, honeycomb panels, and chopped fiber moldings were made and tested. Property data from 219K (-65 F) to 589K (600 F) were obtained. All elements were made in a production environment. The size of each element was sufficient to insure production capability and structural component applicability. Problems associated with adhesive bonding, laminate and structural element analysis, material variability, and test methods were addressed.

  3. Energy Savings Potential and Research, Development, & Demonstration Opportunities for Residential Building Heating, Ventilation, and Air Conditioning Systems

    SciTech Connect

    Goetzler, William; Zogg, Robert; Young, Jim; Schmidt, Justin

    2012-10-01

    This report is an assessment of 135 different heating, ventilation, and air-conditioning (HVAC) technologies for U.S. residential buildings to identify and provide analysis on 19 priority technology options in various stages of development. The analyses include an estimation of technical energy-savings potential, descriptions of technical maturity, descriptions of non-energy benefits, descriptions of current barriers for market adoption, and descriptions of the technology's applicability to different building or HVAC equipment types. From these technology descriptions, are suggestions for potential research, development and demonstration (RD&D) initiatives that would support further development of the priority technology options.

  4. Energy Savings Potential and Research, Development, & Demonstration Opportunities for Commercial Building Heating, Ventilation, and Air Conditioning Systems

    SciTech Connect

    none,

    2011-09-01

    This report covers an assessment of 182 different heating, ventilation, and air-conditioning (HVAC) technologies for U.S. commercial buildings to identify and provide analysis on 17 priority technology options in various stages of development. The analyses include an estimation of technical energy-savings potential, description of technical maturity, description of non-energy benefits, description of current barriers for market adoption, and description of the technology’s applicability to different building or HVAC equipment types. From these technology descriptions, are suggestions for potential research, development and demonstration (RD&D) initiatives that would support further development of the priority technology options.

  5. Regulation of Transfer Functions by the imp Locus of the Streptomyces coelicolor Plasmidogenic Element SLP1

    PubMed Central

    Hagege, Juliette M.; Brasch, Michael A.; Cohen, Stanley N.

    1999-01-01

    SLP1int is a 17.2-kb genetic element that normally is integrated site specifically into the chromosome of Streptomyces coelicolor A3(2). The imp operon within SLP1int represses replication of both chromosomally integrated and extrachromosomal SLP1. During mating with S. lividans, SLP1int can excise, delete part of imp, and form a family of autonomously replicating conjugative plasmids. Earlier work has shown that impA and impC gene products act in concert to control plasmid maintenance and regulate their own transcription. Here we report that these imp genes act also on a second promoter, Popimp (promoter opposite imp), located adjacent to, and initiating transcription divergent from, imp to regulate loci involved in the intramycelial transfer of SLP1 plasmids. spdB1 and spdB2, two overlapping genes immediately 3′ to Popimp and directly regulated by imp, are shown by Tn5 mutagenesis to control transfer-associated growth inhibition (i.e., pocking). Additional genes resembling transfer genes of other Streptomyces spp. plasmids and required for SLP1 transfer and/or postconjugal intramycelial spread are located more distally to Popimp. Expression of impA and impC in an otherwise competent recipient strain prevented SLP1-mediated gene transfer of chromosomal and plasmid genes but not plasmid-independent chromosome-mobilizing activity, suggesting that information transduced to recipients after the formation of mating pairs affects imp activity. Taken together with earlier evidence that the imp operon regulates SLP1 DNA replication, the results reported here implicate imp in the overall regulation of functions related to the extrachromosomal state of SLP1. PMID:10498709

  6. Structural analysis of the catalytic mechanism and stereoselectivity in Streptomyces coelicolor alditol oxidase.

    PubMed

    Forneris, Federico; Heuts, Dominic P H M; Delvecchio, Manuela; Rovida, Stefano; Fraaije, Marco W; Mattevi, Andrea

    2008-01-22

    Alditol oxidase (AldO) from Streptomyces coelicolor A3(2) is a soluble monomeric flavin-dependent oxidase that performs selective oxidation of the terminal primary hydroxyl group of several alditols. Here, we report the crystal structure of the recombinant enzyme in its native state and in complex with both six-carbon (mannitol and sorbitol) and five-carbon substrates (xylitol). AldO shares the same folding topology of the members of the vanillyl-alcohol oxidase family of flavoenzymes and exhibits a covalently linked FAD which is located at the bottom of a funnel-shaped pocket that forms the active site. The high resolution of the three-dimensional structures highlights a well-defined hydrogen-bonding network that tightly constrains the substrate in the productive conformation for catalysis. Substrate binding occurs through a lock-and-key mechanism and does not induce conformational changes with respect to the ligand-free protein. A network of charged residues is proposed to favor catalysis through stabilization of the deprotonated form of the substrate. A His side chain acts as back door that "pushes" the substrate-reactive carbon atom toward the N5-C4a locus of the flavin. Analysis of the three-dimensional structure reveals possible pathways for diffusion of molecular oxygen and a small cavity on the re side of the flavin that may host oxygen during FAD reoxidation. These features combined with the tight shape of the catalytic site provide insights into the mechanism of AldO-mediated regioselective oxidation reactions and its substrate specificity.

  7. Extracellular complementation and the identification of additional genes involved in aerial mycelium formation in Streptomyces coelicolor.

    PubMed

    Nodwell, J R; Yang, M; Kuo, D; Losick, R

    1999-02-01

    Morphogenesis in the bacterium Streptomyces coelicolor involves the formation of a lawn of hair-like aerial hyphae on the colony surface that stands up in the air and differentiates into chains of spores. bld mutants are defective in the formation of this aerial mycelium and grow as smooth, hairless colonies. When certain pairs of bld mutants are grown close to one another on rich sporulation medium, they exhibit extracellular complementation such that one mutant restores aerial mycelium formation to the other. The extracellular complementation relationships of most of the previously isolated bld mutants placed them in a hierarchy of extracellular complementation groups. We have screened for further bld mutants with precautions intended to maximize the discovery of additional genes. Most of the 50 newly isolated mutant strains occupy one of three of the previously described positions in the hierarchy, behaving like bldK, bldC, or bldD mutants. We show that the mutations in some of the strains that behave like bldK are bldK alleles but that others fall in a cluster at a position on the chromosome distinct from that of any known bld gene. We name this locus bldL. By introducing cloned genes into the strains that exhibit bldC or bldD-like extracellular complementation phenotypes, we show that most of these strains are likely to contain mutations in genes other than bldC or bldD. These results indicate that the genetic control of aerial mycelium formation is more complex than previously recognized and support the idea that a high proportion of bld genes are directly or indirectly involved in the production of substances that are exchanged between cells during morphological differentiation.

  8. Extracellular complementation and the identification of additional genes involved in aerial mycelium formation in Streptomyces coelicolor.

    PubMed Central

    Nodwell, J R; Yang, M; Kuo, D; Losick, R

    1999-01-01

    Morphogenesis in the bacterium Streptomyces coelicolor involves the formation of a lawn of hair-like aerial hyphae on the colony surface that stands up in the air and differentiates into chains of spores. bld mutants are defective in the formation of this aerial mycelium and grow as smooth, hairless colonies. When certain pairs of bld mutants are grown close to one another on rich sporulation medium, they exhibit extracellular complementation such that one mutant restores aerial mycelium formation to the other. The extracellular complementation relationships of most of the previously isolated bld mutants placed them in a hierarchy of extracellular complementation groups. We have screened for further bld mutants with precautions intended to maximize the discovery of additional genes. Most of the 50 newly isolated mutant strains occupy one of three of the previously described positions in the hierarchy, behaving like bldK, bldC, or bldD mutants. We show that the mutations in some of the strains that behave like bldK are bldK alleles but that others fall in a cluster at a position on the chromosome distinct from that of any known bld gene. We name this locus bldL. By introducing cloned genes into the strains that exhibit bldC or bldD-like extracellular complementation phenotypes, we show that most of these strains are likely to contain mutations in genes other than bldC or bldD. These results indicate that the genetic control of aerial mycelium formation is more complex than previously recognized and support the idea that a high proportion of bld genes are directly or indirectly involved in the production of substances that are exchanged between cells during morphological differentiation. PMID:9927452

  9. Genome-wide transcriptome analysis reveals that a pleiotropic antibiotic regulator, AfsS, modulates nutritional stress response in Streptomyces coelicolor A3(2)

    PubMed Central

    Lian, Wei; Jayapal, Karthik P; Charaniya, Salim; Mehra, Sarika; Glod, Frank; Kyung, Yun-Seung; Sherman, David H; Hu, Wei-Shou

    2008-01-01

    Background A small "sigma-like" protein, AfsS, pleiotropically regulates antibiotic biosynthesis in Streptomyces coelicolor. Overexpression of afsS in S. coelicolor and certain related species causes antibiotic stimulatory effects in the host organism. Although recent studies have uncovered some of the upstream events activating this gene, the mechanisms through which this signal is relayed downstream leading to the eventual induction of antibiotic pathways remain unclear. Results In this study, we employed whole-genome DNA microarrays and quantitative PCRs to examine the transcriptome of an afsS disruption mutant that is completely deficient in the production of actinorhodin, a major S. coelicolor antibiotic. The production of undecylprodigiosin, another prominent antibiotic, was, however, perturbed only marginally in the mutant. Principal component analysis of temporal gene expression profiles identified two major gene classes each exhibiting a distinct coordinate differential expression pattern. Surprisingly, nearly 70% of the >117 differentially expressed genes were conspicuously associated with nutrient starvation response, particularly those of phosphate, nitrogen and sulfate. Furthermore, expression profiles of some transcriptional regulators including at least two sigma factors were perturbed in the mutant. In almost every case, the effect of afsS disruption was not observed until the onset of stationary phase. Conclusion Our data suggests a comprehensive role for S. coelicolor AfsS as a master regulator of both antibiotic synthesis and nutritional stress response, reminiscent of alternative sigma factors found in several bacteria. PMID:18230178

  10. Sterol 14alpha-demethylase activity in Streptomyces coelicolor A3(2) is associated with an unusual member of the CYP51 gene family.

    PubMed Central

    Lamb, David C; Fowler, Kay; Kieser, Tobias; Manning, Nigel; Podust, Larissa M; Waterman, Michael R; Kelly, Diane E; Kelly, Steven L

    2002-01-01

    The annotation of the genome sequence of Streptomyces coelicolor A3(2) revealed a cytochrome P450 (CYP) resembling various sterol 14alpha-demethylases (CYP51). The putative CYP open reading frame (SC7E4.20) was cloned with a tetrahistidine tag appended to the C-terminus and expressed in Escherichia coli. Protein purified to electrophoretic homogeneity was observed to bind the 14-methylated sterols lanosterol and 24-methylene-24,25-dihydrolanosterol (24-MDL). Reconstitution experiments with E. coli reductase partners confirmed activity in 14alpha-demethylation for 24-MDL, but not lanosterol. An S. coelicolor A3(2) mutant containing a transposon insertion in the CYP51 gene, which will abolish synthesis of the functional haemoprotein, was isolated as a viable strain, the first time a CYP51 has been identified as non-essential. The role of this CYP in bacteria is intriguing. No sterol product was detected in non-saponifiable cell extracts of the parent S. coelicolor A3(2) strain or of the mutant. S. coelicolor A3(2) CYP51 contains very few of the conserved CYP51 residues and, even though it can catalyse 14alpha-demethylation, it probably has another function in Streptomyces. We propose that it is a member of a new CYP51 subfamily. PMID:12023899

  11. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1988-01-01

    Details three demonstrations for use in chemistry classrooms. Includes: "A Demonstration of Corrosion by Differential Aeration"; "A Simple Demonstration of the Activation Energy Concept"; and "A Boiling Demonstration at Room Temperature." Each description includes equipment, materials, and methods. (CW)

  12. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1988-01-01

    Describes two chemistry demonstrations including a demonstration of chemical inhibition and "The Rayleigh Fountain" which demonstrates the polarity of the water molecule. Provides instructions and explanations for each demonstration. (CW)

  13. Overview of the Development of the Solar Electric Propulsion Technology Demonstration Mission 12.5-kW Hall Thruster

    NASA Technical Reports Server (NTRS)

    Kamhawi, Hani; Huang, Wensheng; Haag, Thomas; Yim, John; Chang, Li; Clayman, Lauren; Herman, Daniel; Shastry, Rohit; Thomas, Robert; Verhey, Timothy; Griffith, Christopher; Myers, James; Williams, George; Mikellides, Ioannis; Hofer, Richard; Polk, James; Goebel, Dan

    2014-01-01

    NASA is developing mission concepts for a solar electric propulsion technology demonstration mission. A number of mission concepts are being evaluated including ambitious missions to near Earth objects. The demonstration of a high-power solar electric propulsion capability is one of the objectives of the candidate missions under consideration. In support of NASA's exploration goals, a number of projects are developing extensible technologies to support NASA's near and long term mission needs. Specifically, the Space Technology Mission Directorate Solar Electric Propulsion Technology Demonstration Mission project is funding the development of a 12.5-kilowatt magnetically shielded Hall thruster system to support future NASA missions. This paper presents the design attributes of the thruster that was collaboratively developed by the NASA Glenn Research Center and the Jet Propulsion Laboratory. The paper provides an overview of the magnetic, plasma, thermal, and structural modeling activities that were carried out in support of the thruster design. The paper also summarizes the results of the functional tests that have been carried out to date. The planned thruster performance, plasma diagnostics (internal and in the plume), thermal, wear, and mechanical tests are outlined.

  14. Overview of the Development of the Solar Electric Propulsion Technology Demonstration Mission 12.5-kW Hall Thruster

    NASA Technical Reports Server (NTRS)

    Kamhawi, Hani; Huang, Wensheng; Haag, Thomas; Yim, John; Chang, Li; Clayman, Lauren; Herman, Daniel; Shastry, Rohit; Thomas, Robert; Verhey, Timothy; Griffith, Christopher; Myers, James; Williams, George; Mikellides, Ioannis; Hofer, Richard; Polk, James; Goebel, Dan

    2014-01-01

    NASA is developing mission concepts for a solar electric propulsion technology demonstration mission. A number of mission concepts are being evaluated including ambitious missions to near Earth objects. The demonstration of a high-power solar electric propulsion capability is one of the objectives of the candidate missions under consideration. In support of NASAs exploration goals, a number of projects are developing extensible technologies to support NASAs near and long term mission needs. Specifically, the Space Technology Mission Directorate Solar Electric Propulsion Technology Demonstration Mission project is funding the development of a 12.5-kW magnetically shielded Hall thruster system to support future NASA missions. This paper presents the design attributes of the thruster that was collaboratively developed by the NASA Glenn Research Center and the Jet Propulsion Laboratory. The paper provides an overview of the magnetic, plasma, thermal, and structural modeling activities that were carried out in support of the thruster design. The paper also summarizes the results of the functional tests that have been carried out to date. The planned thruster performance, plasma diagnostics (internal and in the plume), thermal, wear, and mechanical tests are outlined.

  15. Development and Evaluation of Sensor Concepts for Ageless Aerospace Vehicles: Report 3 - Design of the Concept Demonstrator

    NASA Technical Reports Server (NTRS)

    Abbott, David; Ables, Jon; Batten, Adam; Carpenter, David; Collings, Tony; Doyle, Briony; Dunlop, John; Edwards, Graeme; Farmer, Tony; Gaffney, Bruce; Hedley, Mark; Isaacs, Peter; Johnson, Mark; Joshi, Bhautik; Lewis, Chris; Poilton, Geoff; Price, Don; Prokopenko, Mikhail; Reda, Torsten; Rees, David; Scott, Andrew; Seneviratne, Sarath; Valencia, Philip; Wang, Peter; Whitnall, Denis

    2008-01-01

    This report provides an outline of the essential features of a Structural Health Monitoring Concept Demonstrator (CD) that will be constructed during the next eight months. It is emphasized that the design cannot be considered to be complete, and that design work will continue in parallel with construction and testing. A major advantage of the modular design is that small modules of the system can be developed, tested and modified before a commitment is made to full system development. The CD is expected to develop and evolve for a number of years after its initial construction. This first stage will, of necessity, be relatively simple and have limited capabilities. Later developments will improve all aspects of the functionality of the system, including sensing, processing, communications, intelligence and response. The report indicates the directions this later development will take.

  16. Framework Programmable Platform for the Advanced Software Development Workstation (FPP/ASDW). Demonstration framework document. Volume 2: Framework process description

    NASA Technical Reports Server (NTRS)

    Mayer, Richard J.; Blinn, Thomas M.; Dewitte, Paula S.; Crump, John W.; Ackley, Keith A.

    1992-01-01

    In the second volume of the Demonstration Framework Document, the graphical representation of the demonstration framework is given. This second document was created to facilitate the reading and comprehension of the demonstration framework. It is designed to be viewed in parallel with Section 4.2 of the first volume to help give a picture of the relationships between the UOB's (Unit of Behavior) of the model. The model is quite large and the design team felt that this form of presentation would make it easier for the reader to get a feel for the processes described in this document. The IDEF3 (Process Description Capture Method) diagrams of the processes of an Information System Development are presented. Volume 1 describes the processes and the agents involved with each process, while this volume graphically shows the precedence relationships among the processes.

  17. Deletion of the hypothetical protein SCO2127 of Streptomyces coelicolor allowed identification of a new regulator of actinorhodin production.

    PubMed

    H, Tierrafría Víctor; Cuauhtemoc, Licona-Cassani; Nidia, Maldonado-Carmona; Alba, Romero-Rodríguez; Sara, Centeno-Leija; Esteban, Marcellin; Romina, Rodríguez-Sanoja; Ruiz-Villafán, Beatriz; K, Nielsen Lars; Sergio, Sánchez

    2016-11-01

    Although the specific function of SCO2127 remains elusive, it has been assumed that this hypothetical protein plays an important role in carbon catabolite regulation and therefore in antibiotic biosynthesis in Streptomyces coelicolor. To shed light on the functional relationship of SCO2127 to the biosynthesis of actinorhodin, a detailed analysis of the proteins differentially produced between the strain M145 and the Δsco2127 mutant of S. coelicolor was performed. The delayed morphological differentiation and impaired production of actinorhodin showed by the deletion strain were accompanied by increased abundance of gluconeogenic enzymes, as well as downregulation of both glycolysis and acetyl-CoA carboxylase. Repression of mycothiol biosynthetic enzymes was further observed in the absence of SCO2127, in addition to upregulation of hydroxyectoine biosynthetic enzymes and SCO0204, which controls nitrite formation. The data generated in this study reveal that the response regulator SCO0204 greatly contributes to prevent the formation of actinorhodin in the ∆sco2127 mutant, likely through the activation of some proteins associated with oxidative stress that include the nitrite producer SCO0216.

  18. Cloning and analysis of a gene cluster from Streptomyces coelicolor that causes accelerated aerial mycelium formation in Streptomyces lividans.

    PubMed Central

    Ma, H; Kendall, K

    1994-01-01

    We describe the cloning and analysis of two overlapping DNA fragments from Streptomyces coelicolor that cause aerial mycelium to appear more rapidly than usual when introduced into Streptomyces lividans on a low-copy-number plasmid vector. Colonies of S. lividans TK64 harboring either clone produce visible aerial mycelia after only 48 h of growth, rather than the usual 72 to 96 h. From deletion and sequence analysis, this rapid aerial mycelium (Ram) phenotype appears to be due to a cluster of three genes that we have designated ramA, ramB, and ramR. Both ramA and ramB potentially encode 65-kDa proteins with homology to ATP-dependent membrane-translocating proteins. A chromosomal ramB disruption mutant of S. lividans was found to be severely defective in aerial mycelium formation. ramR could encode a 21-kDa protein with significant homology to the UhpA subset of bacterial two-component response regulator proteins. The overall organization and potential proteins encoded by the cloned DNA suggest that this is the S. coelicolor homolog of the amf gene cluster that has been shown to be important for aerial mycelium formation in Streptomyces griseus. However, despite the fact that the two regions probably have identical functions, there is relatively poor homology between the two gene clusters at the DNA sequence level. Images PMID:8206859

  19. ScbR- and ScbR2-mediated signal transduction networks coordinate complex physiological responses in Streptomyces coelicolor.

    PubMed

    Li, Xiao; Wang, Juan; Li, Shanshan; Ji, Junjie; Wang, Weishan; Yang, Keqian

    2015-10-07

    In model organism Streptomyces coelicolor, γ-butyrolactones (GBLs) and antibiotics were recognized as signalling molecules playing fundamental roles in intra- and interspecies communications. To dissect the GBL and antibiotic signalling networks systematically, the in vivo targets of their respective receptors ScbR and ScbR2 were identified on a genome scale by ChIP-seq. These identified targets encompass many that are known to play important roles in diverse cellular processes (e.g. gap1, pyk2, afsK, nagE2, cdaR, cprA, cprB, absA1, actII-orf4, redZ, atrA, rpsL and sigR), and they formed regulatory cascades, sub-networks and feedforward loops to elaborately control key metabolite processes, including primary and secondary metabolism, morphological differentiation and stress response. Moreover, interplay among ScbR, ScbR2 and other regulators revealed intricate cross talks between signalling pathways triggered by GBLs, antibiotics, nutrient availability and stress. Our work provides a global view on the specific responses that could be triggered by GBL and antibiotic signals in S. coelicolor, among which the main echo was the change of production profile of endogenous antibiotics and antibiotic signals manifested a role to enhance bacterial stress tolerance as well, shedding new light on GBL and antibiotic signalling networks widespread among streptomycetes.

  20. A decision analysis framework to support long-term planning for nuclear fuel cycle technology research, development, demonstration and deployment

    SciTech Connect

    Sowder, A.G.; Machiels, A.J.; Dykes, A.A.; Johnson, D.H.

    2013-07-01

    To address challenges and gaps in nuclear fuel cycle option assessment and to support research, develop and demonstration programs oriented toward commercial deployment, EPRI (Electric Power Research Institute) is seeking to develop and maintain an independent analysis and assessment capability by building a suite of assessment tools based on a platform of software, simplified relationships, and explicit decision-making and evaluation guidelines. As a demonstration of the decision-support framework, EPRI examines a relatively near-term fuel cycle option, i.e., use of reactor-grade mixed-oxide fuel (MOX) in U.S. light water reactors. The results appear as a list of significant concerns (like cooling of spent fuels, criticality risk...) that have to be taken into account for the final decision.

  1. Research, development and demonstration of nickel-iron batteries for electric vehicle propulsion. Annual report for 1979

    SciTech Connect

    Not Available

    1980-06-01

    Research progress in the development of Ni/Fe batteries (electrodes in particular) for the period is described. The negative plate demonstrated a reliable lifetime of almost 1000 cycles; 20 mm positive plates were proved feasible; prototype cells yielded output at about 50 Wh/kg and 100 Wh/liter; program goals of 20% greater than these figures appear feasible. 27 figures, 20 tables. (RWR)

  2. Induction of actinorhodin production by rpsL (encoding ribosomal protein S12) mutations that confer streptomycin resistance in Streptomyces lividans and Streptomyces coelicolor A3(2).

    PubMed Central

    Shima, J; Hesketh, A; Okamoto, S; Kawamoto, S; Ochi, K

    1996-01-01

    A strain of Streptomyces lividans, TK24, was found to produce a pigmented antibiotic, actinorhodin, although S. lividans normally does not produce this antibiotic. Genetic analyses revealed that a streptomycin-resistant mutation str-6 in strain TK24 is responsible for induction of antibiotic synthesis. DNA sequencing showed that str-6 is a point mutation in the rpsL gene encoding ribosomal protein S12, changing Lys-88 to Glu. Gene replacement experiments with the Lys88-->Glu str allele demonstrated unambiguously that the str mutation is alone responsible for the activation of actinorhodin production observed. In contrast, the strA1 mutation, a genetic marker frequently used for crosses, did not restore actinorhodin production and was found to result in an amino acid alteration of Lys-43 to Asn. Induction of actinorhodin production was also detected in strain TK21, which does not harbor the str-6 mutation, when cells were incubated with sufficient streptomycin or tetracycline to reduce the cell's growth rate, and 40 and 3% of streptomycin- or tetracycline-resistant mutants, respectively, derived from strain TK21 produced actinorhodin. Streptomycin-resistant mutations also blocked the inhibitory effects of relA and brgA mutations on antibiotic production, aerial mycelium formation or both. These str mutations changed Lys-88 to Glu or Arg and Arg-86 to His in ribosomal protein S12. The decrease in streptomycin production in relC mutants in Streptomyces griseus could also be abolished completely by introducing streptomycin-resistant mutations, although the impairment in antibiotic production due to bldA (in Streptomyces coelicolor) or afs mutations (in S. griseus) was not eliminated. These results indicate that the onset and extent of secondary metabolism in Streptomyces spp. is significantly controlled by the translational machinery. PMID:8955413

  3. Development and Evaluation of Sensor Concepts for Ageless Aerospace Vehicles: Report 6 - Development and Demonstration of a Self-Organizing Diagnostic System for Structural Health Monitoring

    NASA Technical Reports Server (NTRS)

    Batten, Adam; Edwards, Graeme; Gerasimov, Vadim; Hoschke, Nigel; Isaacs, Peter; Lewis, Chris; Moore, Richard; Oppolzer, Florien; Price, Don; Prokopenko, Mikhail; Scott, Andrew; Wang, Peter

    2010-01-01

    This report describes a significant advance in the capability of the CSIRO/NASA structural health monitoring Concept Demonstrator (CD). The main thrust of the work has been the development of a mobile robotic agent, and the hardware and software modifications and developments required to enable the demonstrator to operate as a single, self-organizing, multi-agent system. This single-robot system is seen as the forerunner of a system in which larger numbers of small robots perform inspection and repair tasks cooperatively, by self-organization. While the goal of demonstrating self-organized damage diagnosis was not fully achieved in the time available, much of the work required for the final element that enables the robot to point the video camera and transmit an image has been completed. A demonstration video of the CD and robotic systems operating will be made and forwarded to NASA.

  4. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1987-01-01

    Presented are three demonstrations for chemical education. The activities include: (1) demonstration of vapor pressure; (2) a multicolored luminol-based chemiluminescence demonstration; and (3) a Charles's Law/Vapor pressure apparatus. (RH)

  5. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1982-01-01

    Three chemistry demonstrations are described: (1) partition coefficients; (2) Rutherford simulation experiment; and (3) demonstration of the powerful oxidizing property of dimanganeseheptoxide. Background information, materials needed, and procedures are provided for each demonstration. (JN)

  6. Reflectance Demonstration.

    ERIC Educational Resources Information Center

    Kowalski, Frank

    1993-01-01

    Presents a demonstration in which a mirror "disappears" upon rotation. The author has used the demonstration with students from fourth grade up through college. Suggestions are given for making the demonstration into a permanent hallway display. (MVL)

  7. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1981-01-01

    Provides procedures for demonstrations: (1) the ferrioxalate actinometer, which demonstrates a photochemical reaction; and (2) the silver mirror, which demonstrates the reduction of a metal salt to the metal and/or the reducing power of sugars. (CS)

  8. Demonstration of Decision Support Tools for Sustainable Development - An Application on Alternative Fuels in the Greater Yellowstone-Teton Region

    SciTech Connect

    Shropshire, D.E.; Cobb, D.A.; Worhach, P.; Jacobson, J.J.; Berrett, S.

    2000-12-30

    The Demonstration of Decision Support Tools for Sustainable Development project integrated the Bechtel/Nexant Industrial Materials Exchange Planner and the Idaho National Engineering and Environmental Laboratory System Dynamic models, demonstrating their capabilities on alternative fuel applications in the Greater Yellowstone-Teton Park system. The combined model, called the Dynamic Industrial Material Exchange, was used on selected test cases in the Greater Yellow Teton Parks region to evaluate economic, environmental, and social implications of alternative fuel applications, and identifying primary and secondary industries. The test cases included looking at compressed natural gas applications in Teton National Park and Jackson, Wyoming, and studying ethanol use in Yellowstone National Park and gateway cities in Montana. With further development, the system could be used to assist decision-makers (local government, planners, vehicle purchasers, and fuel suppliers) in selecting alternative fuels, vehicles, and developing AF infrastructures. The system could become a regional AF market assessment tool that could help decision-makers understand the behavior of the AF market and conditions in which the market would grow. Based on this high level market assessment, investors and decision-makers would become more knowledgeable of the AF market opportunity before developing detailed plans and preparing financial analysis.

  9. Development and Evaluation of Sensor Concepts for Ageless Aerospace Vehicles: Report 5 - Phase 2 Implementation of the Concept Demonstrator

    NASA Technical Reports Server (NTRS)

    Batten, Adam; Dunlop, John; Edwards, Graeme; Farmer, Tony; Gaffney, Bruce; Hedley, Mark; Hoschke, Nigel; Isaacs, Peter; Johnson, Mark; Lewis, Chris; Murdoch, Alex; Poulton, Geoff; Price, Don; Prokopenko, Mikhail; Sharp, Ian; Scott, Andrew; Valencia, Philip; Wang, Peter; Whitnall, Denis

    2009-01-01

    This report describes the second phase of the implementation of the Concept Demonstrator experimental test-bed system containing sensors and processing hardware distributed throughout the structure, which uses multi-agent algorithms to characterize impacts and determine a suitable response to these impacts. This report expands and adds to the report of the first phase implementation. The current status of the system hardware is that all 192 physical cells (32 on each of the 6 hexagonal prism faces) have been constructed, although only four of these presently contain data-acquisition sub-modules to allow them to acquire sensor data. Impact detection.. location and severity have been successfully demonstrated. The software modules for simulating cells and controlling the test-bed are fully operational. although additional functionality will be added over time. The visualization workstation displays additional diagnostic information about the array of cells (both real and simulated) and additional damage information. Local agent algorithms have been developed that demonstrate emergent behavior of the complex multi-agent system, through the formation of impact damage boundaries and impact networks. The system has been shown to operate well for multiple impacts. and to demonstrate robust reconfiguration in the presence of damage to numbers of cells.

  10. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1985-01-01

    Background information (including chemical reactions) and procedures used are provided for (1) three buffer demonstrations and (2) a demonstration of phase transfer catalysis and carbanion formation. (JN)

  11. End-Users' Voice in EHR Selection: Development of a Usability Questionnaire for Demonstrations in Procurement (DPUQ).

    PubMed

    Tyllinen, Mari; Kaipio, Johanna; Lääveri, Tinja; Nieminen, Marko

    2017-01-01

    This paper describes the development of a questionnaire for evaluating usability during EHR system procurement (DPUQ). Established usability questionnaires can be used to gather user feedback after using the systems. However, during procurement, experimenting with real system use is practical only with a limited number of system candidates. There is a need for less resource-demanding usability evaluation in the early stages of procurement in cases with several vendors. DPUQ has been designed for usability evaluation by end-users during special scenario-based vendor demonstrations. The questionnaire includes three sets of questions to be used during and after the vendor demonstration. DPUQ delivers specific usability scores and can be used to compare system candidates in procurement complementing other evaluation methods.

  12. GASIS demonstration

    SciTech Connect

    Vidas, E.H.

    1995-04-01

    A prototype of the GASIS database and retrieval software has been developed and is the subject of this poster session and computer demonstration. The prototype consists of test or preliminary versions of the GASIS Reservoir Data System and Source Directory datasets and the software for query and retrieval. The prototype reservoir database covers the Rocky Mountain region and contains the full GASIS data matrix (all GASIS data elements) that will eventually be included on the CD-ROM. It is populated for development purposes primarily by the information included in the Rocky Mountain Gas Atlas. The software has been developed specifically for GASIS using Foxpro for Windows. The application is an executable file that does not require Foxpro to run. The reservoir database software includes query and retrieval, screen display, report generation, and data export functions. Basic queries by state, basin, or field name will be assisted by scrolling selection lists. A detailed query screen will allow record selection on the basis of any data field, such as depth, cumulative production, or geological age. Logical operators can be applied to any-numeric data element or combination of elements. Screen display includes a {open_quotes}browse{close_quotes} display with one record per row and a detailed single record display. Datasets can be exported in standard formats for manipulation with other software packages. The Source Directory software will allow record retrieval by database type or subject area.

  13. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1988-01-01

    Describes three flame test demonstrations including "Student-Presented Demonstrations on the Colors of Transition Metal Complexes,""A Flame Test Demonstration Device," and "Vivid Flame Tests." Preparation and procedures are discussed. Included in the first demonstration is an evaluation scheme for grading student…

  14. Tested Demonstrations

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1977-01-01

    Three demonstrations are described: paramagnetic properties of Fe(11) and Fe(111), the preparation of polyurethane foam: a lecture demonstration and the electrolysis of water-fuel cell reactions. A small discussion of the concepts demonstrated is included in each demonstration's description. (MR)

  15. Research, development and demonstration of nickel-zinc batteries for electric vehicle propulsion. Annual report, 1979. [70 W/lb

    SciTech Connect

    Not Available

    1980-06-01

    This second annual report under Contract No. 31-109-39-4200 covers the period July 1, 1978 through August 31, 1979. The program demonstrates the feasibility of the nickel-zinc battery for electric vehicle propulsion. The program is divided into seven distinct but highly interactive tasks collectively aimed at the development and commercialization of nickel-zinc technology. These basic technical tasks are separator development, electrode development, product design and analysis, cell/module battery testing, process development, pilot manufacturing, and thermal management. A Quality Assurance Program has also been established. Significant progress has been made in the understanding of separator failure mechanisms, and a generic category of materials has been specified for the 300+ deep discharge (100% DOD) applications. Shape change has been reduced significantly. A methodology has been generated with the resulting hierarchy: cycle life cost, volumetric energy density, peak power at 80% DOD, gravimetric energy density, and sustained power. Generation I design full-sized 400-Ah cells have yielded in excess of 70 W/lb at 80% DOD. Extensive testing of cells, modules, and batteries is done in a minicomputer-based testing facility. The best life attained with electric vehicle-size cell components is 315 cycles at 100% DOD (1.0V cutoff voltage), while four-cell (approx. 6V) module performance has been limited to about 145 deep discharge cycles. The scale-up of processes for production of components and cells has progressed to facilitate component production rates of thousands per month. Progress in the area of thermal management has been significant, with the development of a model that accurately represents heat generation and rejection rates during battery operation. For the balance of the program, cycle life of > 500 has to be demonstrated in modules and full-sized batteries. 40 figures, 19 tables. (RWR)

  16. Development and Demonstration of a New Generation High Efficiency 10kW Stationary Fuel Cell System

    SciTech Connect

    Howell, Thomas Russell

    2013-04-30

    The overall project objective is to develop and demonstrate a polymer electrolyte membrane fuel cell combined heat and power (PEMFC CHP) system that provides the foundation for commercial, mass produced units which achieve over 40% electrical efficiency (fuel to electric conversion) from 50-100% load, greater than 70% overall efficiency (fuel to electric energy + usable waste heat energy conversion), have the potential to achieve 40,000 hours durability on all major process components, and can be produced in high volumes at under $400/kW (revised to $750/kW per 2011 DOE estimates) capital cost.

  17. Diverse control of metabolism and other cellular processes in Streptomyces coelicolor by the PhoP transcription factor: genome-wide identification of in vivo targets

    PubMed Central

    Allenby, Nicholas E. E.; Laing, Emma; Bucca, Giselda; Kierzek, Andrzej M.; Smith, Colin P.

    2012-01-01

    Streptomycetes sense and respond to the stress of phosphate starvation via the two-component PhoR–PhoP signal transduction system. To identify the in vivo targets of PhoP we have undertaken a chromatin-immunoprecipitation-on-microarray analysis of wild-type and phoP mutant cultures and, in parallel, have quantified their transcriptomes. Most (ca. 80%) of the previously in vitro characterized PhoP targets were identified in this study among several hundred other putative novel PhoP targets. In addition to activating genes for phosphate scavenging systems PhoP was shown to target two gene clusters for cell wall/extracellular polymer biosynthesis. Furthermore PhoP was found to repress an unprecedented range of pathways upon entering phosphate limitation including nitrogen assimilation, oxidative phosphorylation, nucleotide biosynthesis and glycogen catabolism. Moreover, PhoP was shown to target many key genes involved in antibiotic production and morphological differentiation, including afsS, atrA, bldA, bldC, bldD, bldK, bldM, cdaR, cdgA, cdgB and scbR-scbA. Intriguingly, in the PhoP-dependent cpk polyketide gene cluster, PhoP accumulates substantially at three specific sites within the giant polyketide synthase-encoding genes. This study suggests that, following phosphate limitation, Streptomyces coelicolor PhoP functions as a ‘master’ regulator, suppressing central metabolism, secondary metabolism and developmental pathways until sufficient phosphate is salvaged to support further growth and, ultimately, morphological development. PMID:22904076

  18. Diverse control of metabolism and other cellular processes in Streptomyces coelicolor by the PhoP transcription factor: genome-wide identification of in vivo targets.

    PubMed

    Allenby, Nicholas E E; Laing, Emma; Bucca, Giselda; Kierzek, Andrzej M; Smith, Colin P

    2012-10-01

    Streptomycetes sense and respond to the stress of phosphate starvation via the two-component PhoR-PhoP signal transduction system. To identify the in vivo targets of PhoP we have undertaken a chromatin-immunoprecipitation-on-microarray analysis of wild-type and phoP mutant cultures and, in parallel, have quantified their transcriptomes. Most (ca. 80%) of the previously in vitro characterized PhoP targets were identified in this study among several hundred other putative novel PhoP targets. In addition to activating genes for phosphate scavenging systems PhoP was shown to target two gene clusters for cell wall/extracellular polymer biosynthesis. Furthermore PhoP was found to repress an unprecedented range of pathways upon entering phosphate limitation including nitrogen assimilation, oxidative phosphorylation, nucleotide biosynthesis and glycogen catabolism. Moreover, PhoP was shown to target many key genes involved in antibiotic production and morphological differentiation, including afsS, atrA, bldA, bldC, bldD, bldK, bldM, cdaR, cdgA, cdgB and scbR-scbA. Intriguingly, in the PhoP-dependent cpk polyketide gene cluster, PhoP accumulates substantially at three specific sites within the giant polyketide synthase-encoding genes. This study suggests that, following phosphate limitation, Streptomyces coelicolor PhoP functions as a 'master' regulator, suppressing central metabolism, secondary metabolism and developmental pathways until sufficient phosphate is salvaged to support further growth and, ultimately, morphological development.

  19. Development of a 15 T Nb3Sn accelerator dipole demonstrator at Fermilab

    SciTech Connect

    Novitski, I.; Andreev, N.; Barzi, E.; Carmichael, J.; Kashikhin, V. V.; Turrioni, D.; Yu, M.; Zlobin, A. V.

    2016-06-01

    Here, a 100 TeV scale Hadron Collider (HC) with a nominal operation field of at least 15 T is being considered for the post-LHC era, which requires using the Nb3Sn technology. Practical demonstration of this field level in an accelerator-quality magnet and substantial reduction of the magnet costs are the key conditions for realization of such a machine. FNAL has started the development of a 15 T Nb3Sn dipole demonstrator for a 100 TeV scale HC. The magnet design is based on 4-layer shell type coils, graded between the inner and outer layers to maximize the performance and reduce the cost. The experience gained during the Nb3Sn magnet R&D is applied to different aspects of the magnet design. This paper describes the magnetic and structural designs and parameters of the 15 T Nb3Sn dipole and the steps towards the demonstration model fabrication.

  20. FY15 Status of Immersion Phased Array Ultrasonic Probe Development and Performance Demonstration Results for Under Sodium Viewing

    SciTech Connect

    Diaz, Aaron A.; Larche, Michael R.; Mathews, Royce; Neill, Kevin J.; Baldwin, David L.; Prowant, Matthew S.; Edwards, Matthew K.; Chamberlin, Clyde E.

    2015-09-01

    This Technical Letter Report (TLR) describes work conducted at the Pacific Northwest National Laboratory (PNNL) during FY 2015 on the under-sodium viewing (USV) PNNL project 58745, Work Package AT-15PN230102. This TLR satisfies PNNL’s M3AT-15PN2301027 milestone, and is focused on summarizing the design, development, and evaluation of a two-dimensional matrix phased-array probe referred to as serial number 3 (SN3). In addition, this TLR also provides the results from a performance demonstration of in-sodium target detection trials at 260°C using a one-dimensional 22-element linear array developed in FY14 and referred to as serial number 2 (SN2).

  1. Development and Demonstration of Grid Integration System for PEVs, ESS, and RE: Cooperative Research and Development Final Report, CRADA Number CRD-13-515

    SciTech Connect

    Markel, Tony

    2016-05-01

    NREL and Ideal Power Converters (IPC) will jointly develop and demonstrate a hybrid power converter system integrating bi-directional electric vehicle charging, photovoltaic generation, and stationary battery storage using IPC's 3-Port Hybrid Converter. The organizations will also jointly investigate synergies in tightly integrating these separate power conversion systems.

  2. FY16 Status of Immersion Phased Array Ultrasonic Probe Development and Performance Demonstration Results for Under Sodium Viewing

    SciTech Connect

    Diaz, Aaron A.; Chamberlin, Clyde E.; Edwards, Matthew K.; Hagge, Tobias J.; Hughes, Michael S.; Larche, Michael R.; Mathews, Royce A.; Neill, Kevin J.; Prowant, Matthew S.

    2016-08-31

    This section of the Joint summary technical letter report (TLR) describes work conducted at the Pacific Northwest National Laboratory (PNNL) during FY 2016 (FY16) on the under-sodium viewing (USV) PNNL project 58745, work package AT-16PN230102. This section of the TLR satisfies PNNL’s M3AT-16PN2301025 milestone and is focused on summarizing the design, development, and evaluation of two different phased-array ultrasonic testing (PA-UT) probe designs—a two-dimensional (2D) matrix phased-array probe, and two one-dimensional (1D) linear array probes, referred to as serial number 4 (SN4) engineering test units (ETUs). The 2D probe is a pulse-echo (PE), 32×2, 64-element matrix phased-array ETU. The 1D probes are 32×1 element linear array ETUs. This TLR also provides the results from a performance demonstration (PD) of in-sodium target detection trials at 260°C using both probe designs. This effort continues the iterative evolution supporting the longer term goal of producing and demonstrating a pre-manufacturing prototype ultrasonic probe that possesses the fundamental performance characteristics necessary to enable the development of a high-temperature sodium-cooled fast reactor (SFR) inspection system for in-sodium detection and imaging.

  3. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1987-01-01

    Provides instructions on conducting four demonstrations for the chemistry classroom. Outlines procedures for demonstrations dealing with coupled oscillations, the evaporation of liquids, thioxanthone sulfone radical anion, and the control of variables and conservation of matter. (TW)

  4. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1978-01-01

    Presents two demonstrations; one on Boyle's Law, to illustrate the gas law and serve as a challenging problem for the students; the other is a modified Color Blind Traffic Light demonstration in which the oscillating reactions were speeded up. (GA)

  5. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1978-01-01

    Two demonstrations are described which are suitable for introductory chemistry classes. The first involves the precipitation of silver, and the second is a demonstration of the relationship between rate constants and equilibrium constants using water and beakers. (BB)

  6. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1990-01-01

    Presented are two demonstrations; "Heat of Solution and Colligative Properties: An Illustration of Enthalpy and Entropy," and "A Vapor Pressure Demonstration." Included are lists of materials and experimental procedures. Apparatus needed are illustrated. (CW)

  7. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1989-01-01

    Presented are two demonstrations including a variation of the iodine clock reaction, and a simple demonstration of refractive index. The materials, procedures, and a discussion of probable results are given for each. (CW)

  8. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1981-01-01

    Presents: (1) a simple demonstration which illustrates the driving force of entropy using the familiar effects of the negative thermal expansion coefficient of rubber; and (2) a demonstration of tetrahedral bonding using soap films. (CS)

  9. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1981-01-01

    Two demonstrations are described: (1) red cabbage and electrolysis of water to bring together acid/base and electrochemical concepts; and (2) a model to demonstrate acid/base conjugate pairs utilizing magnets. (SK)

  10. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1987-01-01

    Describes two classroom chemistry demonstrations which focus on the descriptive chemistry of bromine and iodine. Outlines the chemicals and equipment needed, experimental procedures, and discussion of one demonstration of the oxidation states of bromine and iodine, and another demonstration of the oxidation states of iodine. (TW)

  11. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Sands, Robert; And Others

    1982-01-01

    Procedures for two demonstrations are provided. The solubility of ammonia gas in water is demonstrated by introducing water into a closed can filled with the gas, collapsing the can. The second demonstration relates scale of standard reduction potentials to observed behavior of metals in reactions with hydrogen to produce hydrogen gas. (Author/JN)

  12. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1985-01-01

    List of materials needed, procedures used, and results obtained are provided for two demonstrations. The first is an inexpensive and quick method for demonstrating column chromatography of plant pigments of spinach extract. The second is a demonstration of cathodic protection by impressed current. (JN)

  13. Demonstrating Diffusion

    ERIC Educational Resources Information Center

    Foy, Barry G.

    1977-01-01

    Two demonstrations are described. Materials and instructions for demonstrating movement of molecules into cytoplasm using agar blocks, phenolphthalein, and sodium hydroxide are given. A simple method for demonstrating that the rate of diffusion of a gas is inversely proportional to its molecular weight is also presented. (AJ)

  14. Identification of metE as a Second Target of the sRNA scr5239 in Streptomyces coelicolor

    PubMed Central

    Vockenhuber, Michael-Paul; Heueis, Nona; Suess, Beatrix

    2015-01-01

    While transcriptional regulation of the primary and secondary metabolism of the model organism Streptomyces coelicolor is well studied, little is still known about the role small noncoding RNAs (sRNAs) play in regulating gene expression in this organism. Here, we report the identification of a second target of the sRNA scr5239, an sRNA highly conserved in streptomycetes. The 159 nt long sRNA binds its target, the mRNA of the cobalamin independent methionine synthase metE (SCO0985), at the 5’ end of its open reading frame thereby repressing translation. We show that a high methionine level induces expression of scr5239 itself. This leads, in a negative feedback loop, to the repression of methionine biosynthesis. In contrast to the first reported target of this sRNA, the agarase dagA, this interaction seems to be conserved in a wide number of streptomycetes. PMID:25785836

  15. Alteration of coenzyme specificity of malate dehydrogenase from Streptomyces coelicolor A3(2) by site-directed mutagenesis.

    PubMed

    Ge, Y D; Song, P; Cao, Z Y; Wang, P; Zhu, G P

    2014-07-29

    We describe here for the first time the alteration of coenzyme specificity of malate dehydrogenase (MDH) from Streptomyces coelicolor A3(2) (ScMDH). In the present study, we replaced four amino acid residues in the Rossmann fold (βB-αC) region of NADH-dependent ScMDH by site-directed mutagenesis with those of NADPH-dependent MDH (Glu42Gly, Ile43Ser, Pro45Arg, and Ala46Ser). The coenzyme specificity of the mutant enzyme (ScMDH-T4) was examined. Coenzyme specificity of ScMDH-T4 was shifted 2231.3-fold toward NADPH using kcat/Km(coenzyme) as the measurement of coenzyme specificity. Accordingly, the effect of the replacements on coenzyme specificity is discussed. Our work provides further insight into the coenzyme specificity of ScMDH.

  16. Structure and conformational stability of the enzyme I of Streptomyces coelicolor explored by FTIR and circular dichroism.

    PubMed

    Hurtado-Gómez, Estefanía; Barrera, Francisco N; Neira, José L

    2005-04-01

    The bacterial phosphoenolpyruvate (PEP): sugar phosphotransferase system (PTS), formed by a cascade of several proteins, couples the translocation and phosphorylation of specific sugars across cell membranes. The structure and thermal stability of the first protein (enzyme I, EI) of the PTS in Streptomyces coelicolor is studied by using far-UV circular dichroism (CD) and Fourier transform infrared spectroscopy (FTIR) at pH 7.0. The deconvolution of FTIR spectra indicates that the protein is mainly composed by a 35% of alpha-helical structure and 30% of beta-sheet. The thermal denaturation curves, as followed by both techniques, show only a midpoint at 330 K. This thermal denaturation behaviour is different to that observed in other members of the EI family.

  17. [Ca2+ -dependent modulation of antibiotic resistance in Streptomyces lividans 66 and Streptomyces coelicolor A3(2)].

    PubMed

    Bekker, O B; Elizarov, S M; Alekseeva, M T; Liubimova, I K; Danilenko, V N

    2008-01-01

    The level of resistance to antibiotics of various chemical structure in actinobacteria of the genus Streptomyces is shown to be regulated by Ca2+ ions. The inhibitors of Ca2+/calmodulin and Ca2+/phospholipid-dependent serine/threonine protein kinases (STPK) are found to reduce antibiotic resistance of actinobacteria. The effect of Ca2+ -dependent phosphorylation on the activity of the enzymatic aminoglycoside phosphotransferase system protecting actinobacteria from aminoglycoside antibiotics was studied. It is shown that inhibitors of Ca2+/calmodulin and Ca2+/phospholipid-dependent STPK reduced the Ca2+ -induced kanamycin resistance in Streptomyces lividans cells transformed by a hybrid plasmid which contained the aminoglycoside phosphotransferase VIII (APHVIII) gene. In S. coelicolor A3(2) cells, the protein kinase PK25 responsible for APHVIII phosphorylation in vitro was identified. It is suggested that STPK play a major role in the regulation of antibiotic resistance in actinobacteria.

  18. Identification of metE as a second target of the sRNA scr5239 in Streptomyces coelicolor.

    PubMed

    Vockenhuber, Michael-Paul; Heueis, Nona; Suess, Beatrix

    2015-01-01

    While transcriptional regulation of the primary and secondary metabolism of the model organism Streptomyces coelicolor is well studied, little is still known about the role small noncoding RNAs (sRNAs) play in regulating gene expression in this organism. Here, we report the identification of a second target of the sRNA scr5239, an sRNA highly conserved in streptomycetes. The 159 nt long sRNA binds its target, the mRNA of the cobalamin independent methionine synthase metE (SCO0985), at the 5' end of its open reading frame thereby repressing translation. We show that a high methionine level induces expression of scr5239 itself. This leads, in a negative feedback loop, to the repression of methionine biosynthesis. In contrast to the first reported target of this sRNA, the agarase dagA, this interaction seems to be conserved in a wide number of streptomycetes.

  19. Affordable Development and Demonstration of a Small Nuclear Thermal Rocket (NTR) Engine and Stage: How Small Is Big Enough?

    NASA Technical Reports Server (NTRS)

    Borowski, Stanley K.; Sefcik, Robert J.; Fittje, James E.; McCurdy, David R.; Qualls, Arthur L.; Schnitzler, Bruce G.; Werner, James E.; Weitzberg, Abraham; Joyner, Claude R.

    2016-01-01

    The Nuclear Thermal Rocket (NTR) derives its energy from fission of uranium-235 atoms contained within fuel elements that comprise the engine's reactor core. It generates high thrust and has a specific impulse potential of approximately 900 specific impulse - a 100 percent increase over today's best chemical rockets. The Nuclear Thermal Propulsion (NTP) project, funded by NASA's Advanced Exploration Systems (AES) program, includes five key task activities: (1) Recapture, demonstration, and validation of heritage graphite composite (GC) fuel (selected as the Lead Fuel option); (2) Engine Conceptual Design; (3) Operating Requirements Definition; (4) Identification of Affordable Options for Ground Testing; and (5) Formulation of an Affordable Development Strategy. During fiscal year (FY) 2014, a preliminary Design Development Test and Evaluation (DDT&E) plan and schedule for NTP development was outlined by the NASA Glenn Research Center (GRC), Department of Energy (DOE) and industry that involved significant system-level demonstration projects that included Ground Technology Demonstration (GTD) tests at the Nevada National Security Site (NNSS), followed by a Flight Technology Demonstration (FTD) mission. To reduce cost for the GTD tests and FTD mission, small NTR engines, in either the 7.5 or 16.5 kilopound-force thrust class, were considered. Both engine options used GC fuel and a common fuel element (FE) design. The small approximately 7.5 kilopound-force criticality-limited engine produces approximately157 thermal megawatts and its core is configured with parallel rows of hexagonal-shaped FEs and tie tubes (TTs) with a FE to TT ratio of approximately 1:1. The larger approximately 16.5 kilopound-force Small Nuclear Rocket Engine (SNRE), developed by Los Alamos National Laboratory (LANL) at the end of the Rover program, produces approximately 367 thermal megawatts and has a FE to TT ratio of approximately 2:1. Although both engines use a common 35-inch (approximately

  20. Isolation and Characterization of EstC, a New Cold-Active Esterase from Streptomyces coelicolor A3(2)

    PubMed Central

    Brault, Guillaume; Shareck, François; Hurtubise, Yves; Lépine, François; Doucet, Nicolas

    2012-01-01

    The genome sequence of Streptomyces coelicolor A3(2) contains more than 50 genes coding for putative lipolytic enzymes. Many studies have shown the capacity of this actinomycete to store important reserves of intracellular triacylglycerols in nutrient depletion situations. In the present study, we used genome mining of S. coelicolor to identify genes coding for putative, non-secreted esterases/lipases. Two genes were cloned and successfully overexpressed in E. coli as His-tagged fusion proteins. One of the recombinant enzymes, EstC, showed interesting cold-active esterase activity with a strong potential for the production of valuable esters. The purified enzyme displayed optimal activity at 35°C and was cold-active with retention of 25% relative activity at 10°C. Its optimal pH was 8.5–9 but the enzyme kept more than 75% of its maximal activity between pH 7.5 and 10. EstC also showed remarkable tolerance over a wide range of pH values, retaining almost full residual activity between pH 6–11. The enzyme was active toward short-chain p-nitrophenyl esters (C2–C12), displaying optimal activity with the valerate (C5) ester (kcat/Km = 737±77 s−1 mM−1). The enzyme was also very active toward short chain triglycerides such as triacetin (C2:0) and tributyrin (C4:0), in addition to showing good primary alcohol and organic solvent tolerance, suggesting it could function as an interesting candidate for organic synthesis of short-chain esters such as flavors. PMID:22396747

  1. Structural and phylogenetic analysis of a conserved actinobacteria-specific protein (ASP1; SCO1997) from Streptomyces coelicolor

    PubMed Central

    Gao, Beile; Sugiman-Marangos, Seiji; Junop, Murray S; Gupta, Radhey S

    2009-01-01

    Background The Actinobacteria phylum represents one of the largest and most diverse groups of bacteria, encompassing many important and well-characterized organisms including Streptomyces, Bifidobacterium, Corynebacterium and Mycobacterium. Members of this phylum are remarkably diverse in terms of life cycle, morphology, physiology and ecology. Recent comparative genomic analysis of 19 actinobacterial species determined that only 5 genes of unknown function uniquely define this large phylum [1]. The cellular functions of these actinobacteria-specific proteins (ASP) are not known. Results Here we report the first characterization of one of the 5 actinobacteria-specific proteins, ASP1 (Gene ID: SCO1997) from Streptomyces coelicolor. The X-ray crystal structure of ASP1 was determined at 2.2 Ǻ. The overall structure of ASP1 retains a similar fold to the large NP-1 family of nucleoside phosphorylase enzymes; however, the function is not related. Further comparative analysis revealed two regions expected to be important for protein function: a central, divalent metal ion binding pore, and a highly conserved elbow shaped helical region at the C-terminus. Sequence analyses revealed that ASP1 is paralogous to another actinobacteria-specific protein ASP2 (SCO1662 from S. coelicolor) and that both proteins likely carry out similar function. Conclusion Our structural data in combination with sequence analysis supports the idea that two of the 5 actinobacteria-specific proteins, ASP1 and ASP2, mediate similar function. This function is predicted to be novel since the structures of these proteins do not match any known protein with or without known function. Our results suggest that this function could involve divalent metal ion binding/transport. PMID:19515238

  2. Isolation and characterization of EstC, a new cold-active esterase from Streptomyces coelicolor A3(2).

    PubMed

    Brault, Guillaume; Shareck, François; Hurtubise, Yves; Lépine, François; Doucet, Nicolas

    2012-01-01

    The genome sequence of Streptomyces coelicolor A3(2) contains more than 50 genes coding for putative lipolytic enzymes. Many studies have shown the capacity of this actinomycete to store important reserves of intracellular triacylglycerols in nutrient depletion situations. In the present study, we used genome mining of S. coelicolor to identify genes coding for putative, non-secreted esterases/lipases. Two genes were cloned and successfully overexpressed in E. coli as His-tagged fusion proteins. One of the recombinant enzymes, EstC, showed interesting cold-active esterase activity with a strong potential for the production of valuable esters. The purified enzyme displayed optimal activity at 35°C and was cold-active with retention of 25% relative activity at 10°C. Its optimal pH was 8.5-9 but the enzyme kept more than 75% of its maximal activity between pH 7.5 and 10. EstC also showed remarkable tolerance over a wide range of pH values, retaining almost full residual activity between pH 6-11. The enzyme was active toward short-chain p-nitrophenyl esters (C2-C12), displaying optimal activity with the valerate (C5) ester (k(cat)/K(m) = 737±77 s(-1) mM(-1)). The enzyme was also very active toward short chain triglycerides such as triacetin (C2:0) and tributyrin (C4:0), in addition to showing good primary alcohol and organic solvent tolerance, suggesting it could function as an interesting candidate for organic synthesis of short-chain esters such as flavors.

  3. Affordable Development and Demonstration of a Small NTR Engine and Stage: A Preliminary NASA, DOE, and Industry Assessment

    NASA Technical Reports Server (NTRS)

    Borowski, Stanley K.; Sefcik, Robert J.; Fittje, James E.; McCurdy, David R.; Qualls, Arthur L.; Schnitzler, Bruce G.; Werner, James E.; Weitzberg, Abraham; Joyner, Claude R.

    2015-01-01

    The Nuclear Thermal Rocket (NTR) represents the next evolutionary step in cryogenic liquid rocket engines. Deriving its energy from fission of uranium-235 atoms contained within fuel elements that comprise the engine's reactor core, the NTR can generate high thrust at a specific impulse of approx. 900 seconds or more - twice that of today's best chemical rockets. In FY'11, as part of the AISP project, NASA proposed a Nuclear Thermal Propulsion (NTP) effort that envisioned two key activities - "Foundational Technology Development" followed by system-level "Technology Demonstrations". Five near-term NTP activities identified for Foundational Technology Development became the basis for the NCPS project started in FY'12 and funded by NASA's AES program. During Phase 1 (FY'12-14), the NCPS project was focused on (1) Recapturing fuel processing techniques and fabricating partial length "heritage" fuel elements for the two candidate fuel forms identified by NASA and the DOE - NERVA graphite "composite" and the uranium dioxide (UO2) in tungsten "cermet". The Phase 1 effort also included: (2) Engine Conceptual Design; (3) Mission Analysis and Requirements Definition; (4) Identification of Affordable Options for Ground Testing; and (5) Formulation of an Affordable and Sustainable NTP Development Strategy. During FY'14, a preliminary plan for DDT&E was outlined by GRC, the DOE and industry for NASA HQ that involved significant system-level demonstration projects that included GTD tests at the NNSS, followed by a FTD mission. To reduce development costs, the GTD and FTD tests use a small, low thrust (approx. 7.5 or 16.5 klbf) engine. Both engines use graphite composite fuel and a "common" fuel element design that is scalable to higher thrust (approx. 25 klbf) engines by increasing the number of elements in a larger diameter core that can produce greater thermal power output. To keep the FTD mission cost down, a simple "1-burn" lunar flyby mission was considered along with

  4. Development of RNAi in a Drosophila endoparasitoid wasp and demonstration of its efficiency in impairing venom protein production.

    PubMed

    Colinet, Dominique; Kremmer, Laurent; Lemauf, Séverine; Rebuf, Christian; Gatti, Jean-Luc; Poirié, Marylène

    2014-04-01

    Endoparasitoid wasps are essential regulators of insect pests in ecosystems as well as important biological control auxiliaries. Traits important for parasitism success, such as the injection of venom proteins at oviposition, have thus been mainly studied. However, identification of the key genes involved among the large number of genes identified was still prevented by the lack of functional approaches. Here, we report the development of RNA interference (RNAi) in Leptopilina boulardi, a figitid endoparasitoid that performs its entire development inside the Drosophila host. Having set up conditions for in vitro development of parasitoid late larval stages or pupae, we first targeted the cinnabar gene by microinjecting double-stranded RNA (dsRNA), leading to its silencing and production of red-eyed individuals. We then demonstrated that expression of the gene encoding LbGAP, a virulence factor found in a high amount in L. boulardi venom, could be specifically and almost completely silenced. Finally, a time-course analysis revealed that LbGAP silencing lasted during the entire lifetime of L. boulardi. This is the first report of the efficient silencing of venom protein-encoding genes in parasitoid wasps. Overall, RNAi opens the way for a large-scale functional analysis of parasitoid venom factors as well as other traits involved in parasitism success and more largely in the biology of these ecologically important organisms.

  5. Weaving the Native Web: Using Social Network Analysis to Demonstrate the Value of a Minority Career Development Program

    PubMed Central

    Buchwald, Dedra; Dick, Rhonda Wiegman

    2011-01-01

    Purpose American Indian and Alaska Native scientists are consistently among the most underrepresented minority groups in health research. The authors used social network analysis (SNA) to evaluate the Native Investigator Development Program (NIDP), a career development program for junior Native researchers established as a collaboration between the University of Washington and the University of Colorado Denver. Method The study focused on 29 trainees and mentors who participated in the NIDP. Data were collected on manuscripts and grant proposals produced by participants from 1998 to 2007. Information on authorship of manuscripts and collaborations on grant applications was used to conduct social network analyses with 3 measures of centrality and 1 measure of network reach. Both visual and quantitative analyses were performed. Results Participants in the NIDP collaborated on 106 manuscripts and 83 grant applications. Although 3 highly connected individuals, with critical and central roles in the program, accounted for much of the richness of the network, both current core faculty and “graduates” of the program were heavily involved in collaborations on manuscripts and grants. Conclusions This study’s innovative application of SNA demonstrates that collaborative relationships can be an important outcome of career development programs for minority investigators, and that an analysis of these relationships can provide a more complete assessment of the value of such programs. PMID:21512364

  6. Development of a real-time aeroperformance analysis technique for the X-29A advanced technology demonstrator

    NASA Technical Reports Server (NTRS)

    Ray, R. J.; Hicks, J. W.; Alexander, R. I.

    1988-01-01

    The X-29A advanced technology demonstrator has shown the practicality and advantages of the capability to compute and display, in real time, aeroperformance flight results. This capability includes the calculation of the in-flight measured drag polar, lift curve, and aircraft specific excess power. From these elements many other types of aeroperformance measurements can be computed and analyzed. The technique can be used to give an immediate postmaneuver assessment of data quality and maneuver technique, thus increasing the productivity of a flight program. A key element of this new method was the concurrent development of a real-time in-flight net thrust algorithm, based on the simplified gross thrust method. This net thrust algorithm allows for the direct calculation of total aircraft drag.

  7. Development and Demonstration of a Computational Tool for the Analysis of Particle Vitiation Effects in Hypersonic Propulsion Test Facilities

    NASA Technical Reports Server (NTRS)

    Perkins, Hugh Douglas

    2010-01-01

    In order to improve the understanding of particle vitiation effects in hypersonic propulsion test facilities, a quasi-one dimensional numerical tool was developed to efficiently model reacting particle-gas flows over a wide range of conditions. Features of this code include gas-phase finite-rate kinetics, a global porous-particle combustion model, mass, momentum and energy interactions between phases, and subsonic and supersonic particle drag and heat transfer models. The basic capabilities of this tool were validated against available data or other validated codes. To demonstrate the capabilities of the code a series of computations were performed for a model hypersonic propulsion test facility and scramjet. Parameters studied were simulated flight Mach number, particle size, particle mass fraction and particle material.

  8. Development of a real-time aeroperformance analysis technique for the X-29A advanced technology demonstrator

    NASA Technical Reports Server (NTRS)

    Ray, R. J.; Hicks, J. W.; Alexander, R. I.

    1988-01-01

    The X-29A advanced technology demonstrator has shown the practicality and advantages of the capability to compute and display, in real time, aeroperformance flight results. This capability includes the calculation of the in flight measured drag polar, lift curve, and aircraft specific excess power. From these elements, many other types of aeroperformance measurements can be computed and analyzed. The technique can be used to give an immediate postmaneuver assessment of data quality and maneuver technique, thus increasing the productivity of a flight program. A key element of this new method was the concurrent development of a real-time in flight net thrust algorithm, based on the simplified gross thrust method. This net thrust algorithm allows for the direct calculation of total aircraft drag.

  9. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1983-01-01

    Describes a lecture demonstration of a solid state phase transition using a thermodynamic material which changes state at room temperature. Also describes a demonstration on kinetics using a "Big Bang" (trade mark) calcium carbide cannon. Indicates that the cannon is safe to use. (JN)

  10. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1989-01-01

    Presented are two chemistry demonstrations: (1) an alternative method for the demonstration of the properties of alkali metals, water is added to small amounts of metal; (2) an exploration of the properties of hydrogen, helium, propane, and carbon dioxide using an open trough and candle. (MVL)

  11. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1986-01-01

    Outlines a simple, inexpensive way of demonstrating electroplating using the reaction between nickel ions and copper metal. Explains how to conduct a demonstration of the electrolysis of water by using a colored Na2SO4 solution as the electrolyte so that students can observe the pH changes. (TW)

  12. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1982-01-01

    Three chemistry demonstrations are described: (1) modification of copper catalysis demonstration apparatus; (2) experiments in gas-liquid chromatography with simple gas chromatography at room temperature; and (3) equilibria in silver arsenate-arsenic acid and silver phosphate-phosphoric acid systems. Procedures and materials needed are provided.…

  13. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1980-01-01

    Two demonstrations are described: (1) a variant of preparing purple benzene by phase transfer catalysis with quaternary ammonium salts and potassium permanganate in which crown ethers are used; (2) a corridor or "hallway" demonstration in which unknown molecular models are displayed and prizes awarded to students correctly identifying the…

  14. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1984-01-01

    Background information and procedures are provided for a second part to the dichromate volcano demonstration. The green ash produced during the demonstration is reduced to metal using aluminothermy (Goldschmide process). Also describes suitable light sources and spectroscopes for student observation of emission spectra in lecture halls. (JN)

  15. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1983-01-01

    Free radical chlorination of methane is used in organic chemistry to introduce free radical/chain reactions. In spite of its common occurrence, demonstrations of the reaction are uncommon. Therefore, such a demonstration is provided, including background information, preparation of reactants/reaction vessel, introduction of reactants, irradiation,…

  16. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1983-01-01

    Discusses a supplement to the "water to rose" demonstration in which a pink color is produced. Also discusses blood buffer demonstrations, including hydrolysis of sodium bicarbonate, simulated blood buffer, metabolic acidosis, natural compensation of metabolic acidosis, metabolic alkalosis, acidosis treatment, and alkalosis treatment. Procedures…

  17. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1990-01-01

    Presented are three demonstrations: "The Construction and Use of Commercial Voltaic Cell Displays in Freshman Chemistry"; Dramatizing Isotopes: Deuterated Ice Cubes Sink"; and "A Simple Apparatus to Demonstrate Differing Gas Diffusion Rates (Graham's Law)." Materials, procedures, and safety considerations are discussed. (CW)

  18. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1987-01-01

    Describes two laboratory demonstrations in chemistry. One uses dry ice, freon, and freezer bags to demonstrate volume changes, vapor-liquid equilibrium, a simulation of a rain forest, and vaporization. The other uses the clock reaction technique to illustrate fast reactions and kinetic problems in releasing carbon dioxide during respiration. (TW)

  19. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1980-01-01

    Described is a demonstration utilized to measure the heat of vaporization using the Clausius-Clapeyron equation. Explained is that when measurement is made as part of a demonstration, it raises student's consciousness that chemistry is experimentally based. (Author/DS)

  20. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1985-01-01

    Two demonstrations are described. The first (useful as an introduction to kinetics) shows how the rate of a reaction is fast at first and then gradually decreases to zero when one reactant has been used up. The second is a gas density demonstration using 1,1,2-trichloro-1,2,2-trifluoro ethane. (JN)

  1. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1986-01-01

    Provides three descriptions of demonstrations used in various chemistry courses. Includes the use of a simple demonstration model to illustrate principles of chromatography, techniques for using balloons to teach about the behavior of gases, and the use of small concentrations of synthetic polyelectrolytes to induce the flocculation hydrophobic…

  2. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1983-01-01

    Provides directions for setup and performance of two demonstrations. The first demonstrates the principles of Raoult's Law; using a simple apparatus designed to measure vapor pressure. The second illustrates the energy available from alcohol combustion (includes safety precautions) using an alcohol-fueled missile. (JM)

  3. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1989-01-01

    Provided are two demonstrations for an introductory course in chemistry. The first one emphasizes the observation and the interpretation of facts to form hypotheses during the heating of a beaker of water. The second demonstration shows the liquid phase of carbon dioxide using dry ice and a pressure gauge. (YP)

  4. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1986-01-01

    Discusses the photochromic behavior of mercury(II) bis(dithizonate) in providing a colorful demonstration of the effect that visible light can have on the conformation and bonding of molecules in solution. Provides a description of the demonstration itself, along with the preparation needed to complete it. (TW)

  5. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1979-01-01

    Presents two demonstrations for classroom use related to precipitation of ferrous hydroxide and to variation of vapor pressure with temperature. The former demonstration is simple and useful when discussing solubility of ionic compounds electrode potential of transition elements, and mixed valence compounds. (Author/SA)

  6. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1986-01-01

    Describes two demonstrations designed to help chemistry students visualize certain chemical properties. One experiment uses balloons to illustrate the behavior of gases under varying temperatures and pressures. The other uses a makeshift pea shooter and a commercial model to demonstrate atomic structure and the behavior of high-speed particles.…

  7. Enhanced Bank of Kalman Filters Developed and Demonstrated for In-Flight Aircraft Engine Sensor Fault Diagnostics

    NASA Technical Reports Server (NTRS)

    Kobayashi, Takahisa; Simon, Donald L.

    2005-01-01

    In-flight sensor fault detection and isolation (FDI) is critical to maintaining reliable engine operation during flight. The aircraft engine control system, which computes control commands on the basis of sensor measurements, operates the propulsion systems at the demanded conditions. Any undetected sensor faults, therefore, may cause the control system to drive the engine into an undesirable operating condition. It is critical to detect and isolate failed sensors as soon as possible so that such scenarios can be avoided. A challenging issue in developing reliable sensor FDI systems is to make them robust to changes in engine operating characteristics due to degradation with usage and other faults that can occur during flight. A sensor FDI system that cannot appropriately account for such scenarios may result in false alarms, missed detections, or misclassifications when such faults do occur. To address this issue, an enhanced bank of Kalman filters was developed, and its performance and robustness were demonstrated in a simulation environment. The bank of filters is composed of m + 1 Kalman filters, where m is the number of sensors being used by the control system and, thus, in need of monitoring. Each Kalman filter is designed on the basis of a unique fault hypothesis so that it will be able to maintain its performance if a particular fault scenario, hypothesized by that particular filter, takes place.

  8. Scaled Vitrification System III (SVS III) Process Development and Laboratory Tests at the West Valley Demonstration Project

    SciTech Connect

    V. Jain; S. M. Barnes; B. G. Bindi; R. A. Palmer

    2000-04-30

    At the West Valley Demonstration Project (WVDP),the Vitrification Facility (VF)is designed to convert the high-level radioactive waste (HLW)stored on the site to a stable glass for disposal at a Department of Energy (DOE)-specified federal repository. The Scaled Vitrification System III (SVS-III)verification tests were conducted between February 1995 and August 1995 as a supplemental means to support the vitrification process flowsheet, but at only one seventh the scale.During these tests,the process flowsheet was refined and optimized. The SVS-III test series was conducted with a focus on confirming the applicability of the Redox Forecasting Model, which was based on the Index of Feed Oxidation (IFO)developed during the Functional and Checkout Testing of Systems (FACTS)and SVS-I tests. Additional goals were to investigate the prototypical feed preparation cycle and test the new target glass composition. Included in this report are the basis and current designs of the major components of the Scale Vitrification System and the results of the SVS-III tests.The major subsystems described are the feed preparation and delivery, melter, and off-gas treatment systems. In addition,the correlation between the melter's operation and its various parameters;which included feed rate,cold cap coverage,oxygen reduction (redox)state of the glass,melter power,plenum temperature,and airlift analysis;were developed.

  9. Florida-focused climate change lesson demonstrations from the ASK Florida global and regional climate change professional development workshops

    NASA Astrophysics Data System (ADS)

    Weihs, R. R.

    2013-12-01

    A variety of Florida-focused climate change activities will be featured as part of the ASK Florida global and regional climate change professional development workshops. In a combined effort from Florida State University's Center for Ocean-Atmospheric Prediction Studies (COAPS) and University of South Florida's Coalition for Science Literacy (CSL), and supported by NASA's NICE initiative, the ASK Florida professional development workshops are a series of workshops designed to enhance and support climate change information and related pedagogical skills for middle school science teachers from Title-I schools in Florida. These workshops took place during a two-year period from 2011 to 2013 and consisted of two cohorts in Hillsborough and Volusia counties in Florida. Featured activities include lab-style exercises demonstrating topics such as storm surge and coastal geometry, sea level rise from thermal expansion, and the greenhouse effect. These types of labs are modified so that they allow more independent, inquiry thinking as they require teachers to design their own experiment in order to test a hypothesis. Lecture based activities are used to cover a broad range of topics including hurricanes, climate modeling, and sink holes. The more innovative activities are group activities that utilize roll-playing, technology and resources, and group discussion. For example, 'Climate Gallery Walk' is an activity that features group discussions on each of the climate literacy principles established by the United States Global Change Research Program. By observing discussions between individuals and groups, this activity helps the facilitators gather information on their previous knowledge and identify possible misconceptions that will be addressed within the workshops. Furthermore, 'Fact or Misconception' presents the challenge of identifying whether a given statement is fact or misconception based on the material covered throughout the workshops. It serves as a way to

  10. A glgC gene essential only for the first of two spatially distinct phases of glycogen synthesis in Streptomyces coelicolor A3(2).

    PubMed Central

    Martin, M C; Schneider, D; Bruton, C J; Chater, K F; Hardisson, C

    1997-01-01

    By using a PCR approach based on conserved regions of ADP-glucose pyrophosphorylases, a glgC gene was cloned from Streptomyces coelicolor A3(2). The deduced glgC gene product showed end-to-end relatedness to other bacterial ADP-glucose pyrophosphorylases. The glgC gene is about 1,000 kb from the leftmost chromosome end and is not closely linked to either of the two glgB genes of S. coelicolor, which encode glycogen branching enzymes active in different locations in differentiated colonies. Disruption of glgC eliminated only the first of two temporal peaks of ADP-glucose pyrophosphorylase activity and glycogen accumulation and prevented cytologically observable glycogen accumulation in the substrate mycelium of colonies (phase I), while glycogen deposition in young spore chains (phase II) remained readily detectable. The cloned glgC gene therefore encodes an ADP-glucose pyrophosphorylase essential only for phase I (and it is therefore named glgCI). A second, phase II-specific, glgC gene should also exist in S. coelicolor, though it was not detected by hybridization analysis. PMID:9401038

  11. NsrR from Streptomyces coelicolor Is a Nitric Oxide-sensing [4Fe-4S] Cluster Protein with a Specialized Regulatory Function*

    PubMed Central

    Crack, Jason C.; Munnoch, John; Dodd, Erin L.; Knowles, Felicity; Al Bassam, Mahmoud M.; Kamali, Saeed; Holland, Ashley A.; Cramer, Stephen P.; Hamilton, Chris J.; Johnson, Michael K.; Thomson, Andrew J.; Hutchings, Matthew I.; Le Brun, Nick E.

    2015-01-01

    The Rrf2 family transcription factor NsrR controls expression of genes in a wide range of bacteria in response to nitric oxide (NO). The precise form of the NO-sensing module of NsrR is the subject of controversy because NsrR proteins containing either [2Fe-2S] or [4Fe-4S] clusters have been observed previously. Optical, Mössbauer, resonance Raman spectroscopies and native mass spectrometry demonstrate that Streptomyces coelicolor NsrR (ScNsrR), previously reported to contain a [2Fe-2S] cluster, can be isolated containing a [4Fe-4S] cluster. ChIP-seq experiments indicated that the ScNsrR regulon is small, consisting of only hmpA1, hmpA2, and nsrR itself. The hmpA genes encode NO-detoxifying flavohemoglobins, indicating that ScNsrR has a specialized regulatory function focused on NO detoxification and is not a global regulator like some NsrR orthologues. EMSAs and DNase I footprinting showed that the [4Fe-4S] form of ScNsrR binds specifically and tightly to an 11-bp inverted repeat sequence in the promoter regions of the identified target genes and that DNA binding is abolished following reaction with NO. Resonance Raman data were consistent with cluster coordination by three Cys residues and one oxygen-containing residue, and analysis of ScNsrR variants suggested that highly conserved Glu-85 may be the fourth ligand. Finally, we demonstrate that some low molecular weight thiols, but importantly not physiologically relevant thiols, such as cysteine and an analogue of mycothiol, bind weakly to the [4Fe-4S] cluster, and exposure of this bound form to O2 results in cluster conversion to the [2Fe-2S] form, which does not bind to DNA. These data help to account for the observation of [2Fe-2S] forms of NsrR. PMID:25771538

  12. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1986-01-01

    Background information, list of materials needed, and procedures used are provided for a demonstration involving the transformation of a hydrophobic liquid to a partially hydrophobic semisolid. Safety considerations are noted. (JN)

  13. Tested Demonstrations

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1977-01-01

    Describes a room-temperature method for demonstrating phosphorescence by including samples in a polymer matrix. Also discusses the Old Nassau Reaction, a clock reaction which turns orange then black. (MLH)

  14. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Roffia, Sergio; And Others

    1988-01-01

    Reports two electrochemical demonstrations. Uses a hydrogen-oxygen fuel cell to power a clock. Includes description of methods and materials. Investigates the "potato clock" used with different fruits. Lists emf and current for various fruit and electrode combinations. (ML)

  15. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Rehfeld, D. W.; And Others

    1988-01-01

    Describes two demonstrations (1) a dust explosion using a coffee can, candle, rubber tubing, and cornstarch and (2) forming a silicate-polyvinyl alcohol polymer which can be pressed into plastic sheets or molded. Gives specific instructions. (MVL)

  16. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1987-01-01

    Presents three demonstrations suitable for undergraduate chemistry classes. Focuses on experiments with calcium carbide, the induction by iron of the oxidation of iodide by dichromate, and the classical iodine clock reaction. (ML)

  17. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L.

    1990-01-01

    Included are three demonstrations that include the phase change of ice when under pressure, viscoelasticity and colloid systems, and flame tests for metal ions. The materials, procedures, probable results, and applications to real life situations are included. (KR)

  18. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1979-01-01

    Presents a recipe for the Nylon Rope Trick, which is considered to be one of the most spectacular demonstrations in chemistry. Materials for growing the polymer and some safety precautions are given. (SA)

  19. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1988-01-01

    Describes two demonstrations for use in college chemistry classes. Includes "Spectroscopy in Large Lecture Halls" and "The Endothermic Dissolution of Ammonium Nitrate." Gives materials lists and procedures as well as a discussion of the results. (CW)

  20. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L.

    1982-01-01

    Two demonstrations are described: (1) a sunset effect using a gooseneck lamp and 20 sheets of paper and (2) the preparation and determination of structural features of dimethyl sulfoxide (DMSO) by infrared spectroscopy. (SK)

  1. Development and Demonstration of a Modeling Framework for Assessing the Efficacy of Using Mine Water for Thermoelectric Power Generation

    SciTech Connect

    2010-03-01

    Thermoelectric power plants use large volumes of water for condenser cooling and other plant operations. Traditionally, this water has been withdrawn from the cleanest water available in streams and rivers. However, as demand for electrical power increases it places increasing demands on freshwater resources resulting in conflicts with other off stream water users. In July 2002, NETL and the Governor of Pennsylvania called for the use of water from abandoned mines to replace our reliance on the diminishing and sometimes over allocated surface water resource. In previous studies the National Mine Land Reclamation Center (NMLRC) at West Virginia University has demonstrated that mine water has the potential to reduce the capital cost of acquiring cooling water while at the same time improving the efficiency of the cooling process due to the constant water temperatures associated with deep mine discharges. The objectives of this project were to develop and demonstrate a user-friendly computer based design aid for assessing the costs, technical and regulatory aspects and potential environmental benefits for using mine water for thermoelectric generation. The framework provides a systematic process for evaluating the hydrologic, chemical, engineering and environmental factors to be considered in using mine water as an alternative to traditional freshwater supply. A field investigation and case study was conducted for the proposed 300 MW Beech Hollow Power Plant located in Champion, Pennsylvania. The field study based on previous research conducted by NMLRC identified mine water sources sufficient to reliably supply the 2-3,000gpm water supply requirement of Beech Hollow. A water collection, transportation and treatment system was designed around this facility. Using this case study a computer based design aid applicable to large industrial water users was developed utilizing water collection and handling principals derived in the field investigation and during previous

  2. Treatment plan for aqueous/organic/decontamination wastes under the Oak Ridge Reservation FFCA Development, Demonstration, Testing, and Evaluation Program

    SciTech Connect

    Backus, P.M.; Benson, C.E.; Gilbert, V.P.

    1994-08-01

    The U.S. Department of Energy (DOE) Oak Ridge Operations Office and the U.S. Environmental Protection Agency (EPA)-Region IV have entered into a Federal Facility Compliance Agreement (FFCA) which seeks to facilitate the treatment of low-level mixed wastes currently stored at the Oak Ridge Reservation (ORR) in violation of the Resource, Conservation and Recovery Act Land Disposal Restrictions. The FFCA establishes schedules for DOE to identify treatment for wastes, referred to as Appendix B wastes, that current have no identified or existing capacity for treatment. A development, demonstration, testing, and evaluation (DDT&E) program was established to provide the support necessary to identify treatment methods for mixed was meeting the Appendix B criteria. The Program has assembled project teams to address treatment development needs for major categories of the Appendix B wastes based on the waste characteristics and possible treatment technologies. The Aqueous, Organic, and Decontamination (A/O/D) project team was established to identify pretreatment options for aqueous and organic wastes which will render the waste acceptable for treatment in existing waste treatment facilities and to identify the processes to decontaminate heterogeneous debris waste. In addition, the project must also address the treatment of secondary waste generated by other DDT&E projects. This report details the activities to be performed under the A/O/D Project in support of the identification, selection, and evaluation of treatment processes. The goals of this plan are (1) to determine the major aqueous and organic waste streams requiring treatment, (2) to determine the treatment steps necessary to make the aqueous and organic waste acceptable for treatment in existing treatment facilities on the ORR or off-site, and (3) to determine the processes necessary to decontaminate heterogeneous wastes that are considered debris.

  3. Development of simulated tank wastes for the US Department of Energy`s Underground Storage Tank Integrated Demonstration

    SciTech Connect

    Elmore, M.R.; Colton, N.G.; Jones, E.O.

    1992-08-01

    The purpose of the Underground Storage Tank Integrated Demonstration (USTID) is to identify and evaluate technologies that may be used to characterize, retrieve, treat, and dispose of hazardous and radioactive wastes contained in tanks on US Department of Energy sites. Simulated wastes are an essential component of the evaluation process because they provide controlled samples for technology assessment, and minimize costs and risks involved when working with radioactive wastes. Pacific Northwest Laboratory has developed a recipe to simulate Hanford single-shell tank, (SST) waste. The recipe is derived from existing process recipes, and elemental concentrations are based on characterization data from 18 SSTs. In this procedure, salt cake and metal oxide/hydroxide sludge are prepared individually, and mixed together at varying ratios depending on the specific tank, waste to be simulated or the test being conducted. Elemental and physical properties of the stimulant are comparable with analyzed tank samples, and chemical speciation in the simulant is being improved as speciation data for actual wastes become available. The nonradioactive chemical waste simulant described here is useful for testing technologies on a small scale.

  4. Development of a DC Glow Discharge Exhibit for the Demonstration of Plasma Behavior in a Magnetic Field

    NASA Astrophysics Data System (ADS)

    Bruder, Daniel

    2010-11-01

    The DC Glow Discharge Exhibit is intended to demonstrate the effects a magnetic field produces on a plasma in a vacuum chamber. The display, which will be featured as a part of The Liberty Science Center's ``Energy Quest Exhibition,'' consists of a DC glow discharge tube and information panels to educate the general public on plasma and its relation to fusion energy. Wall posters and an information booklet will offer brief descriptions of fusion-based science and technology, and will portray plasma's role in the development of fusion as a viable source of energy. The display features a horse-shoe magnet on a movable track, allowing viewers to witness the effects of a magnetic field upon a plasma. The plasma is created from air within a vacuum averaging between 100-200 mTorr. Signage within the casing describes the hardware components. The display is pending delivery to The Liberty Science Center, and will replace a similar, older exhibit presently at the museum.

  5. Soils, surficial geology, and geomorphology of the Bear Creek Valley Low-Level Waste Disposal Development and Demonstration Program site

    SciTech Connect

    Lietzke, D.A.; Lee, S.Y.; Lambert, R.E.

    1988-04-01

    An intensive soil survey was conducted on the proposed Low-Level Waste Disposal Development and Demonstration Program site (LLWDDD) in Bear Creek Valley. Soils on the site were related to the underlying residuum and to the surficial colluvium and alluvium. Within any particular geologic formation, soils were subdivided based mostly on the degree of weathering, as reflected by saprolite weathering and morphologic features of the soils. Degree of weathering was related both to slope shape and gradient and to the joint-fracture system. Erosion classes were also used to make further subdivisions of any particular soil. Deep pits were dug in each of the major Conasauga Group formations (Pumpkin Valley, Rogersville, Maryville, and Nolichucky) for soil and saprolite characterization. Because of the widespread presence of alluvium and colluvium, which are potential sources of fill and final cover material, pits and trenches were dug to characterize the properties of these soils and to try to understand the past geomorphic history of the site. The results of the soil survey investigation indicated that the deeply weathered Pumpkin Valley residuum has good potential for the construction of tumuli or other types of belowground or aboveground burial of prepackaged compacted waste. 11 refs., 30 figs., 3 tabs.

  6. Developing and demonstrating low-energy climate control and production techniques for greenhouse-grown citrus and ornamental crops

    SciTech Connect

    Bodnaruk, W.H. Jr.

    1983-04-01

    The aim of this study was to develop and demonstrate low energy climate control and production techniques for greenhouse grown citrus and ornamental crops. Emphasis was placed on design, fuel efficiency and plant response to warm water soil heating systems using solar energy and LP gas. An energy requirement of 28Btus output per hour per square foot of bed space will provide soil temperature of 70/sup 0/F minimum when air temperatures are maintained at 60/sup 0/F. Soil heating to 70/sup 0/ increased rooting and growth of 8 foliage plant varieties by 25 to 45% compared to plants grown under 60/sup 0/F air temperature conditions. Providing soil heating, however, increased fuel consumption in the central Florida test facilities by 30% in the winters of 1980-81 and 1981-82. Solar tie-in to soil heating systems has the potential of reducing fuel usage. Solar heated water provided 4 hours of soil heating following a good collection day. Decreased in-bed pipe spacing and increased storage capacity should increase the solar percentage to 6 hours.

  7. Develop and demonstrate manufacturing processes for fabricating graphite filament reinforced polymide (Gr/PI) composite structural elements

    NASA Technical Reports Server (NTRS)

    Chase, V. A.; Harrison, E. S.

    1985-01-01

    A study was conducted to assess the merits of using graphite/polyimide, NR-150B2 resin, for structural applications on advanced space launch vehicles. The program was divided into two phases: (1) Fabrication Process Development; and (2) Demonstration Components. The first phase of the program involved the selection of a graphite fiber, quality assurance of the NR-150B2 polyimide resin, and the quality assurance of the graphite/polyimide prepreg. In the second phase of the program, a limited number of components were fabricated before the NR-150B2 resin system was removed from the market by the supplier, Du Pont. The advancement of the NR-150B2 polyimide resin binder was found to vary significantly based on previous time and temperature history during the prepregging operation. Strength retention at 316C (600F) was found to be 50% that of room temperature strength. However, the composite would retain its initial strength after 200 hours exposure at 316C (600F). Basic chemistry studies are required for determining NR-150B2 resin binder quality assurance parameters. Graphite fibers are available that can withstand high temperature cure and postcure cycles.

  8. Demonstration Explosion

    NASA Astrophysics Data System (ADS)

    Lee, Charles "Skip"

    1998-05-01

    Last week I did a demonstration that produced a serious explosion. After putting methanol in a big glass carboy and rotating the carboy to build up some methanol vapor, I lit the mouth of the carboy. What normally happens is a "jet engine" effect out of the mouth of the carboy. In my case, the carboy exploded. Two polycarbonate blast shields were shattered and glass was blown as far as 15 feet away. I was not seriously cut and bruised, but had I not been using the two blast shields, I would have been severely injured. At this time, I am not sure what caused the explosion. I have done this demonstration around one hundred times with no problem using the exact same amount of methanol and technique. I think it is important to get the word out that this demonstration may be more dangerous than previously thought. I would also welcome any hypotheses concerning what caused the carboy to explode.

  9. Office of Technology Development FY 1993 program summary: Office of Research and Development, Office of Demonstration, Testing and Evaluation. Revision 1

    SciTech Connect

    Not Available

    1994-02-01

    This report summarizes significant FY93 programmatic information and accomplishments relevant to the individual activities within the Office of Technology Development Program for Research, Development, Demonstration, Testing, and Evaluation (RDDT&E). A brief discussion of the mission of the Office of Environmental Restoration and Waste Management (EM) and the Office of Technology Development is presented. An overview is presented of the major problem areas confronting DOE. These problem areas include: groundwater and soils cleanup; waste retrieval and processing; and pollution prevention. The organizational elements within EM are highlighted. An EM-50 Funding Summary for FY92 and FY93 is also provided. RDDT&E programs are discussed and their key problem areas are summarized. Three salient program-formulating concepts are explained. They are: Integrated Demonstrations, Integrated Programs, and the technology window of opportunity. Detailed information for each of the programs within RDDT&E is presented and includes a fact sheet, a list of technical task plans and an accomplishments and objectives section.

  10. Research, development and demonstration of a fuel cell/battery powered bus system. Phase 1, Final report

    SciTech Connect

    1990-02-28

    Purpose of the Phase I effort was to demonstrate feasibility of the fuel cell/battery system for powering a small bus (under 30 ft or 9 m) on an urban bus route. A brassboard powerplant was specified, designed, fabricated, and tested to demonstrate feasibility in the laboratory. The proof-of-concept bus, with a powerplant scaled up from the brassboard, will be demonstrated under Phase II.

  11. Polydiglycosylphosphate Transferase PdtA (SCO2578) of Streptomyces coelicolor A3(2) Is Crucial for Proper Sporulation and Apical Tip Extension under Stress Conditions

    PubMed Central

    Sigle, Steffen; Steblau, Nadja; Wohlleben, Wolfgang

    2016-01-01

    ABSTRACT Although anionic glycopolymers are crucial components of the Gram-positive cell envelope, the relevance of anionic glycopolymers for vegetative growth and morphological differentiation of Streptomyces coelicolor A3(2) is unknown. Here, we show that the LytR-CpsA-Psr (LCP) protein PdtA (SCO2578), a TagV-like glycopolymer transferase, has a dual function in the S. coelicolor A3(2) life cycle. Despite the presence of 10 additional LCP homologs, PdtA is crucial for proper sporulation. The integrity of the spore envelope was severely affected in a pdtA deletion mutant, resulting in 34% nonviable spores. pdtA deletion caused a significant reduction in the polydiglycosylphosphate content of the spore envelope. Beyond that, apical tip extension and normal branching of vegetative mycelium were severely impaired on high-salt medium. This growth defect coincided with the mislocalization of peptidoglycan synthesis. Thus, PdtA itself or the polydiglycosylphosphate attached to the peptidoglycan by the glycopolymer transferase PdtA also has a crucial function in apical tip extension of vegetative hyphae under stress conditions. IMPORTANCE Anionic glycopolymers are underappreciated components of the Gram-positive cell envelope. They provide rigidity to the cell wall and position extracellular enzymes involved in peptidoglycan remodeling. Although Streptomyces coelicolor A3(2), the model organism for bacterial antibiotic production, is known to produce two distinct cell wall-linked glycopolymers, teichulosonic acid and polydiglycosylphosphate, the role of these glycopolymers in the S. coelicolor A3(2) life cycle has not been addressed so far. This study reveals a crucial function of the anionic glycopolymer polydiglycosylphosphate for the growth and morphological differentiation of S. coelicolor A3(2). Polydiglycosylphosphate is attached to the spore wall by the LytR-CpsA-Psr protein PdtA (SCO2578), a component of the Streptomyces spore wall-synthesizing complex (SSSC), to

  12. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1985-01-01

    Describes two demonstrations that require almost no preparation time, are visually stimulating, and present a variety of material for class discussion (with sample questions provided). The first involves a sodium bicarbonate hydrochloric acid volcano; the second involves a dissolving polystyrene cup. Procedures used and information on…

  13. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1987-01-01

    Describes two demonstrations to illustrate characteristics of substances. Outlines a method to detect the changes in pH levels during the electrolysis of water. Uses water pistols, one filled with methane gas and the other filled with water, to illustrate the differences in these two substances. (TW)

  14. Tested Demonstrations

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1976-01-01

    Describes two demonstrations: one that illustrates the attainment of equilibrium in first-order reactions by changing the volumes of two beakers of water at a specified rate, and another that illustrates the role of indicators in showing pH changes in buffer solutions. (MLH)

  15. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1989-01-01

    Discusses three broad classes of magnetic behavior: diamagnetic, paramagnetic, and ferromagnetic. Presents a simple lecture demonstration using an overhead projector to synthesize triiron tetraoxide and to show its interaction with a magnetic field and comparing it to a paramagnetic material. (MVL)

  16. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1981-01-01

    Provides instructions and a list of materials needed to demonstrate: (1) a model of the quantum mechanical atom; (2) principles involved in metal corrosion and in the prevention of this destructive process by electrochemical means; and (3) a Thermit reaction, modified to make it more dramatic and interesting for students. (SK)

  17. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L.

    1983-01-01

    An apparatus is described in which effects of pressure, volume, and temperature changes on a gas can be observed simultaneously. Includes use of the apparatus in demonstrating Boyle's, Gay-Lussac's, and Charles' Laws, attractive forces, Dalton's Law of Partial pressures, and in illustrating measurable vapor pressures of liquids and some solids.…

  18. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1984-01-01

    Two demonstrations are described. The first shows the effect of polarity on solubility. The second is based on the unexpected formation of a precipitate of barium nitrate when barium carbonate or barium phosphate is treated with dilute nitric acid. List of materials needed and procedures used are included. (JN)

  19. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1985-01-01

    Background information, procedures, and typical results obtained are provided for two demonstrations. The first involves the colorful complexes of copper(II). The second involves reverse-phase separation of Food, Drug, and Cosmetic (FD & C) dyes using a solvent gradient. (JN)

  20. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Cliche, Jean-Marie; And Others

    1988-01-01

    Describes two demonstrations: 1) the effect of polarity on solubility using sodium dichromate, TTE, ligroin, and water to form nonpolar-polar-nonpolar layers with the polar layer being colored; 2) determination of egg whites to be yellow by determining the content of yellow colored riboflavin in the egg white. (MVL)

  1. Development and Demonstration of a Self-Calibrating Pseudolite Array for Task Level Control of a Planetary Rover

    NASA Technical Reports Server (NTRS)

    Rock, Stephen M.; LeMaster, Edward A.

    2001-01-01

    Pseudolites can extend the availability of GPS-type positioning systems to a wide range of applications not possible with satellite-only GPS. One such application is Mars exploration, where the centimeter-level accuracy and high repeatability of CDGPS would make it attractive for rover positioning during autonomous exploration, sample collection, and habitat construction if it were available. Pseudolites distributed on the surface would allow multiple rovers and/or astronauts to share a common navigational reference. This would help enable cooperation for complicated science tasks, reducing the need for instructions from Earth and increasing the likelihood of mission success. Conventional GPS Pseudolite arrays require that the devices be pre-calibrated through a Survey of their locations, typically to sub-centimeter accuracy. This is a problematic task for robots on the surface of another planet. By using the GPS signals that the Pseudolites broadcast, however, it is possible to have the array self-survey its own relative locations, creating a SelfCalibrating Pseudolite Array (SCPA). This requires the use of GPS transceivers instead of standard pseudolites. Surveying can be done either at carrier- or code-phase levels. An overview of SCPA capabilities, system requirements, and self-calibration algorithms is presented in another work. The Aerospace Robotics Laboratory at Statif0id has developed a fully operational prototype SCPA. The array is able to determine the range between any two transceivers with either code- or carrier-phase accuracy, and uses this inter-transceiver ranging to determine the at-ray geometry. This paper presents results from field tests conducted at Stanford University demonstrating the accuracy of inter-transceiver ranging and its viability and utility for array localization, and shows how transceiver motion may be utilized to refine the array estimate by accurately determining carrier-phase integers and line biases. It also summarizes the

  2. DOE FY 2010 Budget Request and Recovery Act Funding for Energy Research, Development, Demonstration, and Deployment: Analysis and Recommendations

    SciTech Connect

    Anadon, Laura Diaz; Gallagher, Kelly Sims; Bunn, Matthew

    2009-06-01

    -term. Energy storage may play a crucial role in the future of the power and transportation systems, which together consume two thirds of primary energy in the United States. A recent National Academy of Science report recommended carrying out detailed scenario assessments of the penetration of unconventional fuels from coal and coal and biomass with CCS. And the research plan provided for nuclear fission does not justify spending as many funds as were requested. The proposed funding for FY 2010 and the resources from ARRA, however, do not guarantee that the United States will finally enjoy the predictable and consistent publicly-funded energy technology innovation effort that it needs. The Obama administration must put in place a comprehensive energy technology innovation strategy that will ensure that an expanded ERD3 effort is both sustainable and efficient. This commission would be charged with, inter alia, developing a strategy that optimizes the integration of the various stages of innovation (research, development, demonstration, early deployment), as well as integrates efforts across technology areas. The database upon which this analysis is based may be downloaded in Excel format at: http://belfercenter.ksg.harvard.edu/publication/19119/ .

  3. ENVIRONMENTAL RESEARCH BRIEF: DEVELOPMENT OF GAS CLEANING TECHNOLOGY: DEMONSTRATION OF ADVANCED ELECTROSTATIC PRECIPITATOR TECHNOLOGY (INDIA ESP TRAINING)

    EPA Science Inventory

    The Brief discusses a demonstration of advanced electrostatic precipitator (ESP) diagnostics and technologies in India. Six Indian ESP specialists were selected by Southern Research Institute and their consultants, with the concurrence of EPA's project officer, to attend a course...

  4. SpaceFibre Demonstrator (Demonstration and Testing)

    NASA Astrophysics Data System (ADS)

    Helfers, T.; Rastetter, P.; Papadas, C.; Parkes, S.

    2014-08-01

    Currently Astrium GmbH and ISD S.A. are planning the development of a demonstrator for SpaceFibre. The SpaceFibre demonstrator will be used to execute functional performance tests and EMC (Electro Magnetic Compatibility) tests. University of Dundee is program prime contractor and provides Astrium with the SpaceFibre IP core. The work si shared between the two partners in the following way: • Astrium: Prime Contractor and Technical Coordination; FPGA Design; EMC Testing• ISD: Development of Demonstrator Board including housing, development of test bed and functional performance testingThe driving requirements for this development are:• SpaceFibre performance, while implementing it into space equivalent components• Design and MAIT of the demonstrator in such a way that representative EMC testing is possible.

  5. A Guide for Creating Community Awareness and Developing Interagency Cooperation. Preschool Program: A Regional Demonstration Program for Preschool Handicapped Children.

    ERIC Educational Resources Information Center

    Eagen, Carol S.; And Others

    One of 10 documents developed for preschool programs for handicapped children, the manual focuses on a transdisciplinary approach to developing community awareness and interagency cooperation. The program provides concrete methods for helping an agency understand a community's character and develop a tailor made awareness program. An introductory…

  6. Demonstration of Nondeclarative Sequence Learning in Mice: Development of an Animal Analog of the Human Serial Reaction Time Task

    ERIC Educational Resources Information Center

    Christie, Michael A.; Hersch, Steven M.

    2004-01-01

    In this paper, we demonstrate nondeclarative sequence learning in mice using an animal analog of the human serial reaction time task (SRT) that uses a within-group comparison of behavior in response to a repeating sequence versus a random sequence. Ten female B6CBA mice performed eleven 96-trial sessions containing 24 repetitions of a 4-trial…

  7. ACTION FOR BOSTON COMMUNITY DEVELOPMENT, INC. AND BRANDEIS UNIVERSITY DEMONSTRATION COLLEGE COMPUS PROGRAM, SUMMER, 1964 BRANDIS UNIVERSITY, WALTHAM, MASSACHUSETTES.

    ERIC Educational Resources Information Center

    SACHAR, ABRAHAM

    A LARGE PERCENTAGE OF BOYS FROM POORER URBAN NEIGHBORHOODS DO NOT SEEK EDUCATION BEYOND HIGH SCHOOL, FOR FINANCIAL REASONS OR LACK OF INTEREST IN OR KNOWLEDGE ABOUT COLLEGE LIFE. THIS PROPOSAL HAS AS ITS PRIMARY OBJECTIVES TO DEMONSTRATE THAT POTENTIALLY ABLE BUT UNDERACHIEVING STUDENTS, WHEN EXPOSED TO A SUMMER PROGRAM THAT RECOGNIZES AND…

  8. Identification of a Novel Lincomycin Resistance Mutation Associated with Activation of Antibiotic Production in Streptomyces coelicolor A3(2).

    PubMed

    Wang, Guojun; Izawa, Masumi; Yang, Xiaoge; Xu, Dongbo; Tanaka, Yukinori; Ochi, Kozo

    2017-02-01

    Comparative genome sequencing analysis of a lincomycin-resistant strain of Streptomyces coelicolor A3(2) and the wild-type strain identified a novel mutation conferring a high level of lincomycin resistance. Surprisingly, the new mutation was an in-frame DNA deletion in the genes SCO4597 and SCO4598, resulting in formation of the hybrid gene linR. SCO4597 and SCO4598 encode two histidine kinases, which together with SCO4596, encoding a response regulator, constitute a unique two-component system. Sequence analysis indicated that these three genes and their arrangement patterns are ubiquitous among all Streptomyces genomes sequenced to date, suggesting these genes play important regulatory roles. Gene replacement showed that this mutation was responsible for the high level of lincomycin resistance, the overproduction of the antibiotic actinorhodin, and the enhanced morphological differentiation of this strain. Moreover, heterologous expression of the hybrid gene linR in Escherichia coli conferred resistance to lincomycin in this organism. Introduction of the hybrid gene linR in various Streptomyces strains by gene engineering technology may widely activate and/or enhance antibiotic production.

  9. The chaplins: a family of hydrophobic cell-surface proteins involved in aerial mycelium formation in Streptomyces coelicolor

    PubMed Central

    Elliot, Marie A.; Karoonuthaisiri, Nitsara; Huang, Jianqiang; Bibb, Maureen J.; Cohen, Stanley N.; Kao, Camilla M.; Buttner, Mark J.

    2003-01-01

    The filamentous bacterium Streptomyces coelicolor differentiates by forming specialized, spore-bearing aerial hyphae that grow into the air. Using microarrays, we identified genes that are down-regulated in a mutant unable to erect aerial hyphae. Through this route, we identified a previously unknown layer of aerial mycelium surface proteins (the “chaplins”). The chaplins share a hydrophobic domain of ∼40 residues (the “chaplin domain”), and all have a secretion signal. The five short chaplins (ChpD,E,F,G,H) have one chaplin domain, whereas the three long chaplins (ChpA,B,C) have two chaplin domains and a C-terminal “sorting signal” that targets them for covalent attachment to the cell wall by sortase enzyme. Expression of the two chaplin genes examined (chpE, chpH) depended on aerial hyphae formation but not sporulation, and egfp fusions showed their expression localized to aerial structures. Mass spectrometry of cell wall extracts confirmed that the short chaplins localized to the cell surface. Deletion of chaplin genes caused severe delays in aerial hyphae formation, a phenotype rescued by exogenous application of chaplin proteins. These observations implicate the chaplins in aerial mycelium formation, and suggest that coating of the envelope by the chaplins is required for aerial hyphae to grow out of the aqueous environment of the substrate mycelium into the air. PMID:12832397

  10. Identification and biochemical characterization of a thermostable malate dehydrogenase from the mesophile Streptomyces coelicolor A3(2).

    PubMed

    Ge, Ya-Dong; Cao, Zheng-Yu; Wang, Zong-Da; Chen, Lu-Lu; Zhu, You-Ming; Zhu, Guo-Ping

    2010-01-01

    We identified and characterized a malate dehydrogenase from Streptomyces coelicolor A3(2) (ScMDH). The molecular mass of ScMDH was 73,353.5 Da with two 36,675.0 Da subunits as analyzed by matrix-assisted laser-desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS). The detailed kinetic parameters of recombinant ScMDH are reported here. Heat inactivation studies showed that ScMDH was more thermostable than most MDHs from other organisms, except for a few extremely thermophile bacteria. Recombinant ScMDH was highly NAD(+)-specific and displayed about 400-fold (k(cat)) and 1,050-fold (k(cat)/K(m)) preferences for oxaloacetate reduction over malate oxidation. Substrate inhibition studies showed that ScMDH activity was inhibited by excess oxaloacetate (K(i)=5.8 mM) and excess L-malate (K(i)=12.8 mM). Moreover, ScMDH activity was not affected by most metal ions, but was strongly inhibited by Fe(2+) and Zn(2+). Taken together, our findings indicate that ScMDH is significantly thermostable and presents a remarkably high catalytic efficiency for malate synthesis.

  11. Cosmid based mutagenesis causes genetic instability in Streptomyces coelicolor, as shown by targeting of the lipoprotein signal peptidase gene

    PubMed Central

    Munnoch, John T.; Widdick, David A.; Chandra, Govind; Sutcliffe, Iain C.; Palmer, Tracy; Hutchings, Matthew I.

    2016-01-01

    Bacterial lipoproteins are extracellular proteins tethered to cell membranes by covalently attached lipids. Deleting the lipoprotein signal peptidase (lsp) gene in Streptomyces coelicolor results in growth and developmental defects that cannot be restored by reintroducing lsp. This led us to hypothesise that lsp is essential and that the lsp mutant we isolated previously had acquired compensatory secondary mutations. Here we report resequencing of the genomes of wild-type M145 and the cis-complemented ∆lsp mutant (BJT1004) to map and identify these secondary mutations but we show that they do not increase the efficiency of disrupting lsp and are not lsp suppressors. We provide evidence that they are induced by introducing the cosmid St4A10∆lsp, as part of ReDirect PCR mutagenesis protocol, which transiently duplicates a number of important cell division genes. Disruption of lsp using a suicide vector (which does not result in gene duplication) still results in growth and developmental delays and we conclude that loss of Lsp function results in developmental defects due to the loss of all lipoproteins from the cell membrane. Significantly, our results also indicate the use of cosmid libraries for the genetic manipulation of bacteria can lead to phenotypes not necessarily linked to the gene(s) of interest. PMID:27404047

  12. Influence of transition metals on Streptomyces coelicolor and S. sioyaensis and generation of chromate-reducing mutants.

    PubMed

    Gren, Tetiana; Ostash, Bohdan; Hrubskyy, Yaroslav; Tistechok, Stepan; Fedorenko, Victor

    2014-03-01

    Bacteria-assisted bioremediation is widely recognized as a low-cost method to minimize the consequences of soil pollution with toxic metals originating from industrial sites. Strains used in bioremediation have to deal with high metal load via biosorption, reduction, bioprecipitation, metal sequestration, and/or chelation. Actinobacteria, and streptomycetes in particular, are considered a perspective group for bioremediation as natural soil inhabitants with extensive secondary metabolism. Nevertheless, there is no reference information on survival of the model streptomycetes in the presence of the most abundant metal pollutants. Also, there are no reports describing the selection approaches towards improvement of bioremediation properties. In this work, the resistance of Streptomyces coelicolor M145 and Streptomyces sioyaensis Lv81 to certain transition metals and their growth under different pH values are described for the first time. Spontaneous chromate-resistant S. sioyaensis Lv81-138 strain was selected in the course of this work. Strain Lv81-138 is the most efficient actinobacterial Cr(VI) reducer reported so far, capable of converting 12 mmol/L of Cr(VI) into Cr(III) in a medium supplemented with 50 mmol/L K2CrO4.

  13. LAL Regulators SCO0877 and SCO7173 as Pleiotropic Modulators of Phosphate Starvation Response and Actinorhodin Biosynthesis in Streptomyces coelicolor

    PubMed Central

    Guerra, Susana M.; Rodríguez-García, Antonio; Santos-Aberturas, Javier; Vicente, Cláudia M.; Payero, Tamara D.; Martín, Juan F.; Aparicio, Jesús F.

    2012-01-01

    LAL regulators (Large ATP-binding regulators of the LuxR family) constitute a poorly studied family of transcriptional regulators. Several regulators of this class have been identified in antibiotic and other secondary metabolite gene clusters from actinomycetes, thus they have been considered pathway-specific regulators. In this study we have obtained two disruption mutants of LAL genes from S. coelicolor (Δ0877 and Δ7173). Both mutants were deficient in the production of the polyketide antibiotic actinorhodin, and antibiotic production was restored upon gene complementation of the mutants. The use of whole-genome DNA microarrays and quantitative PCRs enabled the analysis of the transcriptome of both mutants in comparison with the wild type. Our results indicate that the LAL regulators under study act globally affecting various cellular processes, and amongst them the phosphate starvation response and the biosynthesis of the blue-pigmented antibiotic actinorhodin. Both regulators act as negative modulators of the expression of the two-component phoRP system and as positive regulators of actinorhodin biosynthesis. To our knowledge this is the first characterization of LAL regulators with wide implications in Streptomyces metabolism. PMID:22363654

  14. METHODS DEVELOPMENT AND DEMONSTRATION FOR ASSESSING LEVEL OF NATIVE PESTICIDES, PCBS, PAHS, AND VOCS IN HUMAN BLOOD

    EPA Science Inventory

    Methods have been developed for screening and assessing the level of volatile, semi-volatile and non-volatile organic pollutants in human blood. The specific methodology is developed for measuring the presence of "native" compounds rather than their metabolites. Spe...

  15. Research, development, and demonstration of nickel-zinc batteries for electric vehicle propulsion. Annual report for 1980

    SciTech Connect

    Not Available

    1981-03-01

    Progress in the development of nickel-zinc batteries for electric vehicles is reported. Information is presented on nickel electrode preparation and testing; zinc electrode preparation with additives and test results; separator development and the evaluation of polymer-blend separator films; sealed Ni-Zn cells; and the optimization of electric vehicle-type Ni-Zn cells. (LCL)

  16. Research, development, and demonstration of nickel-zinc batteries for electric vehicle propulsion. Annual report for 1980

    SciTech Connect

    Not Available

    1981-03-01

    Progress in developing nickel-zinc batteries for propelling electric vehicles is reported. Information is included on component design, battery fabrication, and module performance testing. Although full scale hardware performance has fallen short of the contract cycle life goals, significant progress has been made to warrant further development. (LCL)

  17. Research, development, and demonstration of nickel-iron batteries for electric vehicle propulsion. Annual report for 1980

    SciTech Connect

    Not Available

    1981-03-01

    The FY 1980 program continued to involve full-size, prototype cell, module and battery fabrication and evaluation, aimed at advancing the technical capabilities of the nickel-iron battery, while simultaneously reducing its potential cost in materials and process areas. Improved Electroprecipitation Process (EPP) nickel electrodes of design thickness (2.5 mm) are now being prepared that display stable capacities of 23 to 25 Ah for the C/3 drain rate at 200+ test cycles. Iron electrodes of the composite-type are delivering 24 Ah at the target thickness (1.0 mm). Iron electrodes are displaying capacity stability for > 1000 test cycles in continuing 3 plate cell tests. Finished cells have delivered 57 to 61 Wh/kg at C/3, and have demonstrated cyclic stability to 500+ cycles at 80% depth of discharge profiles at Westinghouse. A 6-cell module that demonstrated 239 Ah, 1735 Wh, 48 Wh/kg at the C/3 drain rate has also been evaluated at the National Battery Test Laboratory, ANL. It operated for 327 test cycles, to a level of 161 Ah at the C/3 rate, before being removed from test. Reduction in nickel electrode swelling (and concurrent stack starvation), to improve cycling, continues to be an area of major effort to reach the final battery cycle life objectives. Pasted nickel electrodes continue to show promise for meeting the life objectives while, simultaneously, providing a low manufacturing cost. Refinements have occurred in the areas of cell hardware, module manifolding and cell interconnections. These improvements have been incorporated into the construction and testing of the cells and modules for this program. Temperature tests at 0/sup 0/C were performed on a 6-cell module and showed a decrease in capacity of only 25% in Ah and .29% in Wh as compared to 25/sup 0/C performance. Additional tests are planned to demonstrate performance at -15/sup 0/C and 40/sup 0/C.

  18. Combining chitinase C and N-acetylhexosaminidase from Streptomyces coelicolor A3(2) provides an efficient way to synthesize N-acetylglucosamine from crystalline chitin.

    PubMed

    Nguyen-Thi, Nhung; Doucet, Nicolas

    2016-02-20

    The enzymatic bioconversion of chitin is of considerable interest for the natural production of bioactive compounds such as chitooligosaccharides and N-acetyl-d-glucosamine (GlcNAc). Key enzymes are involved in the natural processing of chitin, hydrolyzing this abundant biopolymer to yield chitooligosaccharides with substantial value to the medicinal and biotechnological fields. In this study, chitinase C (ScChiC) from the soil bacterium and chitin decomposer Streptomyces coelicolor A3(2) was expressed, purified and characterized. We also optimized a Streptomyces lividans system generating ScChiC expression yields nearly 500-fold higher than the previously reported heterologous expression in Escherichia coli. The purified enzyme was found to be stable below 55°C for a broad range of pH values (pH 3.5-9) and exhibited high activity against chitin and chitooligosaccharides to form chitobiose (C2) as main product. Crab shell chitin hydrolysis profiles also revealed that ScChiC catalyzes the bioconversion of chitopolysaccharides through an endo-nonprocessive mode of action. When combining ScChiC with an N-acetylhexosaminidase from S. coelicolor A3(2) (ScHEX) in an assay using crude extracts and crystalline chitin as substrate, GlcNAc was generated as final product with a yield over 90% after 8h incubation. This chitin hydrolysis yield represents one of the most efficient enzyme bioconversion of chitopolysaccharides to GlcNAc characterized to date, making the S. coelicolor ScChiC-ScHEX pair a potentially suitable contender for the viable industrial production of this important bioactive compound.

  19. Identification and Biochemical Characterization of Sco3487 from Streptomyces coelicolor A3(2), an Exo- and Endo-Type β-Agarase-Producing Neoagarobiose

    PubMed Central

    Temuujin, Uyangaa; Chi, Won-Jae; Chang, Yong-Keun

    2012-01-01

    Streptomyces coelicolor can degrade agar, the main cell wall component of red macroalgae, for growth. To constitute a crucial carbon source for bacterial growth, the alternating α-(1,3) and β-(1,4) linkages between the 3,6-anhydro-l-galactoses and d-galactoses of agar must be hydrolyzed by α/β-agarases. In S. coelicolor, DagA was confirmed to be an endo-type β-agarase that degrades agar into neoagarotetraose and neoagarohexaose. Genomic sequencing data of S. coelicolor revealed that Sco3487, annotated as a putative hydrolase, has high similarity to the glycoside hydrolase (GH) GH50 β-agarases. Sco3487 encodes a primary translation product (88.5 kDa) of 798 amino acids, including a 45-amino-acid signal peptide. The sco3487 gene was cloned and expressed under the control of the ermE promoter in Streptomyces lividans TK24. β-Agarase activity was detected in transformant culture broth using the artificial chromogenic substrate p-nitrophenyl-β-d-galactopyranoside. Mature Sco3487 (83.9 kDa) was purified 52-fold with a yield of 66% from the culture broth. The optimum pH and temperature for Sco3487 activity were 7.0 and 40°C, respectively. The Km and Vmax for agarose were 4.87 mg/ml (4 × 10−5 M) and 10.75 U/mg, respectively. Sco3487 did not require metal ions for its activity, but severe inhibition by Mn2+ and Cu2+ was observed. Thin-layer chromatography analysis, matrix-assisted laser desorption ionization–time of flight mass spectrometry, and Fourier transform-nuclear magnetic resonance spectrometry of the Sco3487 hydrolysis products revealed that Sco3487 is both an exo- and endo-type β-agarase that degrades agarose, neoagarotetraose, and neoagarohexaose into neoagarobiose. PMID:22020647

  20. The global role of ppGpp synthesis in morphological differentiation and antibiotic production in Streptomyces coelicolor A3(2)

    PubMed Central

    Hesketh, Andrew; Chen, Wenqiong Joan; Ryding, Jamie; Chang, Sherman; Bibb, Mervyn

    2007-01-01

    Background Regulation of production of the translational apparatus via the stringent factor ppGpp in response to amino acid starvation is conserved in many bacteria. However, in addition to this core function, it is clear that ppGpp also exhibits genus-specific regulatory effects. In this study we used Affymetrix GeneChips to more fully characterize the regulatory influence of ppGpp synthesis on the biology of Streptomyces coelicolor A3(2), with emphasis on the control of antibiotic biosynthesis and morphological differentiation. Results Induction of ppGpp synthesis repressed transcription of the major sigma factor hrdB, genes with functions associated with active growth, and six of the thirteen conservons present in the S. coelicolor genome. Genes induced following ppGpp synthesis included the alternative sigma factor SCO4005, many for production of the antibiotics CDA and actinorhodin, the regulatory genes SCO4198 and SCO4336, and two alternative ribosomal proteins. Induction of the CDA and actinorhodin clusters was accompanied by an increase in transcription of the pathway regulators cdaR and actII-ORF4, respectively. Comparison of transcriptome profiles of a relA null strain, M570, incapable of ppGpp synthesis with its parent M600 suggested the occurrence of metabolic stress in the mutant. The failure of M570 to sporulate was associated with a stalling between production of the surfactant peptide SapB, and of the hydrophobins: it overproduced SapB but failed to express the chaplin and rodlin genes. Conclusion In S. coelicolor, ppGpp synthesis influences the expression of several genomic elements that are particularly characteristic of streptomycete biology, notably antibiotic gene clusters, conservons, and morphogenetic proteins. PMID:17683547