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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. 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....

  3. 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

  4. Generalized transduction in Streptomyces coelicolor.

    PubMed

    Burke, J; Schneider, D; Westpheling, J

    2001-05-22

    We report the isolation of generalized transducing phages for Streptomyces species able to transduce chromosomal markers or plasmids between derivatives of Streptomyces coelicolor, the principal genetic model system for this important bacterial genus. We describe four apparently distinct phages (DAH2, DAH4, DAH5, and DAH6) that are capable of transducing multiple chromosomal markers at frequencies ranging from 10(-5) to 10(-9) per plaque-forming unit. The phages contain DNA ranging in size from 93 to 121 kb and mediate linked transfer of genetic loci at neighboring chromosomal sites sufficiently close to be packaged within the same phage particle. The key to our ability to demonstrate transduction by these phages was the establishment of conditions expected to severely reduce superinfection killing during the selection of transductants. The host range of these phages, as measured by the ability to form plaques, extends to species as distantly related as Streptomyces avermitilis and Streptomyces verticillus, which are among the most commercially important species of this genus. Transduction of plasmid DNA between S. coelicolor and S. verticillus was observed at frequencies of approximately 10(-4) transductants per colony-forming unit.

  5. Tryptophan promotes morphological and physiological differentiation in Streptomyces coelicolor.

    PubMed

    Palazzotto, Emilia; Renzone, Giovanni; Fontana, Pietro; Botta, Luigi; Scaloni, Andrea; Puglia, Anna Maria; Gallo, Giuseppe

    2015-12-01

    The molecular mechanisms regulating tryptophan biosynthesis in actinomycetes are poorly understood; similarly, the possible roles of tryptophan in the differentiation program of microorganism life-cycle are still underexplored. To unveil the possible regulatory effect of this amino acid on gene expression, an integrated study based on quantitative teverse transcription-PCR (qRT-PCR) and proteomic approaches was performed on the actinomycete model Streptomyces coelicolor. Comparative analyses on the microorganism growth in a minimal medium with or without tryptophan supplementation showed that biosynthetic trp gene expression in S. coelicolor is not subjected to a negative regulation by the presence of the end product. Conversely, tryptophan specifically induces the transcription of trp genes present in the biosynthetic gene cluster of the calcium-dependent antibiotic (CDA), a lipopeptide containing D- and L-tryptophan residues. In addition, tryptophan stimulates the transcription of the CDA gene cluster regulator cdaR and, coherently, CDA production. Surprisingly, tryptophan also promotes the production of actinorhodin, another antibiotic that does not contain this amino acid in its structure. Combined 2D-DIGE and nano liquid chromatography electrospray linear ion trap tandem mass spectrometry (LC-ESI-LIT-MS/MS) analyses revealed that tryptophan exerts a growth-stage-dependent global effect on S. coelicolor proteome, stimulating anabolic pathways and promoting the accumulation of key factors associated with morphological and physiological differentiation at the late growth stages. Phenotypic observations by scanning electron microscopy and spore production assays demonstrated an increased sporulation in the presence of tryptophan. Transcriptional analysis of catabolic genes kynA and kynB suggested that the actinomycete also uses tryptophan as a carbon and nitrogen source. In conclusion, this study originally provides the molecular basis underlying the stimulatory

  6. Transcriptomics-based strain optimization tool for designing secondary metabolite overproducing strains of Streptomyces coelicolor.

    PubMed

    Kim, Minsuk; Yi, Jeong Sang; Lakshmanan, Meiyappan; Lee, Dong-Yup; Kim, Byung-Gee

    2016-03-01

    In silico model-driven analysis using genome-scale model of metabolism (GEM) has been recognized as a promising method for microbial strain improvement. However, most of the current GEM-based strain design algorithms based on flux balance analysis (FBA) heavily rely on the steady-state and optimality assumptions without considering any regulatory information. Thus, their practical usage is quite limited, especially in its application to secondary metabolites overproduction. In this study, we developed a transcriptomics-based strain optimization tool (tSOT) in order to overcome such limitations by integrating transcriptomic data into GEM. Initially, we evaluated existing algorithms for integrating transcriptomic data into GEM using Streptomyces coelicolor dataset, and identified iMAT algorithm as the only and the best algorithm for characterizing the secondary metabolism of S. coelicolor. Subsequently, we developed tSOT platform where iMAT is adopted to predict the reaction states, and successfully demonstrated its applicability to secondary metabolites overproduction by designing actinorhodin (ACT), a polyketide antibiotic, overproducing strain of S. coelicolor. Mutants overexpressing tSOT targets such as ribulose 5-phosphate 3-epimerase and NADP-dependent malic enzyme showed 2 and 1.8-fold increase in ACT production, thereby validating the tSOT prediction. It is expected that tSOT can be used for solving other metabolic engineering problems which could not be addressed by current strain design algorithms, especially for the secondary metabolite overproductions.

  7. Cross-regulation among disparate antibiotic biosynthetic pathways of Streptomyces coelicolor.

    PubMed

    Huang, Jianqiang; Shi, Jing; Molle, Virginie; Sohlberg, Björn; Weaver, David; Bibb, Maureen J; Karoonuthaisiri, Nitsara; Lih, Chih-Jian; Kao, Camilla M; Buttner, Mark J; Cohen, Stanley N

    2005-12-01

    A complex programme of regulation governs gene expression during development of the morphologically and biochemically complex eubacterial genus Streptomyces. Earlier work has suggested a model in which 'higher level' pleiotropic regulators activate 'pathway-specific' regulators located within chromosomal gene clusters encoding biosynthesis of individual antibiotics. We used mutational analysis and adventitious overexpression of key Streptomyces coelicolor regulators to investigate functional interactions among them. We report here that cluster-situated regulators (CSRs) thought to be pathway-specific can also control other antibiotic biosynthetic gene clusters, and thus have pleiotropic actions. Surprisingly, we also find that CSRs exhibit growth-phase-dependent control over afsR2/afsS, a 'higher level' pleiotropic regulatory locus not located within any of the chromosomal gene clusters it targets, and further demonstrate that cross-regulation by CSRs is modulated globally and differentially during the S. coelicolor growth cycle by the RNaseIII homologue AbsB. Our results, which reveal a network of functional interactions among regulators that govern production of antibiotics and other secondary metabolites in S. coelicolor, suggest that revision of the currently prevalent view of higher-level versus pathway-specific regulation of secondary metabolism in Streptomyces species is warranted.

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

    PubMed

    Piret, J M; Chater, K F

    1985-09-01

    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.

  9. Recombinational cloning of the antibiotic biosynthetic gene clusters in linear plasmid SCP1 of Streptomyces coelicolor A3(2).

    PubMed

    Zhang, Ran; Xia, Haiyang; Xu, Qingyu; Dang, Fujun; Qin, Zhongjun

    2013-08-01

    The model organism Streptomyces coelicolor A3(2) harbors a 356-kb linear plasmid, SCP1. We report here development of a recombinational cloning method for deleting large segment from one telomere of SCP1 followed by replacing with the telomere of pSLA2 and sequentially inserting with the overlapping cosmids in vivo. The procedure depends on homologous recombination coupled with cleavage at telomere termini by telomere terminal protein. Using this procedure, we cloned the 81-kb avermectin and the 76-kb spinosad biosynthetic gene clusters into SCP1. Heterologous expression of avermectin production in S. coelicolor was detected. These results demonstrate the utility of SCP1 for cloning large DNA segments such as antibiotic biosynthetic gene clusters.

  10. 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.

  11. Manipulating and understanding antibiotic production in Streptomyces coelicolor A3(2) with decoy oligonucleotides.

    PubMed

    McArthur, Michael; Bibb, Mervyn J

    2008-01-22

    We have adapted and extended the decoy oligonucleotide technique for use in prokaryotes. To identify cis-acting regulatory elements within a promoter, we developed a DNase I/T7 exonuclease footprinting technique and applied it to actII-orf4 from Streptomyces coelicolor A3(2), which encodes the pathway-specific activator for production of the antibiotic actinorhodin. Our in vivo mapping data allowed us to create decoy oligonucleotides incorporating the identified regulatory elements and to test whether their introduction into S. coelicolor affected antibiotic production. We mapped the promoter region when in a transcriptionally inactive state before the onset of actinorhodin production with the aim of designing decoy oligonucleotides capable of interfering with potential repressor binding and so stimulate actinorhodin production. Mapping identified five candidates for decoy oligonucleotides, and these were tested in a plate-based assay to rapidly validate their activity. A transfection protocol was developed for liquid cultures that enabled efficient uptake of decoys, and quantitative real-time PCR demonstrated decoy persistence for >70 h. Measurement of the effects on growth, expression of actII-orf4, and antibiotic production demonstrated that one of the decoys, in concordance with the plate assay, was more efficacious than the others in increasing actinorhodin production. Two of the identified regulatory elements occurred upstream of gene SCO5812, deletion of which reduced actinorhodin production, confirming that experimental analysis of regulatory motifs can provide new insights into factors influencing antibiotic production in streptomycetes.

  12. 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

  13. Synthetic Promoter Library for Modulation of Actinorhodin Production in Streptomyces coelicolor A3(2)

    PubMed Central

    Sohoni, Sujata Vijay; Fazio, Alessandro; Workman, Christopher T.; Mijakovic, Ivan; Lantz, Anna Eliasson

    2014-01-01

    The objective of this study was the application of the synthetic promoter library (SPL) technology for modulation of actinorhodin production in Streptomyces coelicolor A3(2). The SPL technology was used to optimize the expression of a pathway specific positive transcriptional regulator ActII orf4, which activates the transcription of the S. coelicolor actinorhodin biosynthetic gene cluster. The native actII orf4 promoter was replaced with synthetic promoters, generating a S. coelicolor library with a broad range of expression levels of actII orf4. The resulting library was screened based on the yield of actinorhodin. Selected strains were further physiologically characterized. One of the strains from the library, ScoSPL20, showed considerably higher yield of actinorhodin and final actinorhodin titer, compared to S. coelicolor wild type and S. coelicolor with actII orf4 expressed from a strong constitutive promoter. ScoSPL20 demonstrated exceptional productivity despite having a comparatively weak expression from the promoter. Interestingly, the ScoSPL20 promoter was activated at a much earlier stage of growth compared to the wild type, demonstrating the advantage of fine-tuning and temporal tuning of gene expression in metabolic engineering. Transcriptome studies were performed in exponential and actinorhodin-producing phase of growth to compare gene expression between ScoSPL20 and the wild type. To our knowledge, this is the first successful application of the SPL technology for secondary metabolite production in filamentous bacteria. PMID:24963940

  14. 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.

  15. Genome-wide inference of regulatory networks in Streptomyces coelicolor

    PubMed Central

    2010-01-01

    Background The onset of antibiotics production in Streptomyces species is co-ordinated with differentiation events. An understanding of the genetic circuits that regulate these coupled biological phenomena is essential to discover and engineer the pharmacologically important natural products made by these species. The availability of genomic tools and access to a large warehouse of transcriptome data for the model organism, Streptomyces coelicolor, provides incentive to decipher the intricacies of the regulatory cascades and develop biologically meaningful hypotheses. Results In this study, more than 500 samples of genome-wide temporal transcriptome data, comprising wild-type and more than 25 regulatory gene mutants of Streptomyces coelicolor probed across multiple stress and medium conditions, were investigated. Information based on transcript and functional similarity was used to update a previously-predicted whole-genome operon map and further applied to predict transcriptional networks constituting modules enriched in diverse functions such as secondary metabolism, and sigma factor. The predicted network displays a scale-free architecture with a small-world property observed in many biological networks. The networks were further investigated to identify functionally-relevant modules that exhibit functional coherence and a consensus motif in the promoter elements indicative of DNA-binding elements. Conclusions Despite the enormous experimental as well as computational challenges, a systems approach for integrating diverse genome-scale datasets to elucidate complex regulatory networks is beginning to emerge. We present an integrated analysis of transcriptome data and genomic features to refine a whole-genome operon map and to construct regulatory networks at the cistron level in Streptomyces coelicolor. The functionally-relevant modules identified in this study pose as potential targets for further studies and verification. PMID:20955611

  16. Streptomyces coelicolor SCO4226 is a nickel binding protein.

    PubMed

    Lu, Mo; Jiang, Yong-Liang; Wang, Shu; Jin, Hua; Zhang, Rong-Guang; Virolle, Marie-Joelle; Chen, Yuxing; Zhou, Cong-Zhao

    2014-01-01

    The open reading frame SCO4226 of Streptomyces coelicolor A3(2) encodes an 82-residue hypothetical protein. Biochemical assays revealed that each SCO4226 dimer binds four nickel ions. To decipher the molecular function, we solved the crystal structures of SCO4226 in both apo- and nickel-bound (Ni-SCO4226) forms at 1.30 and 2.04 Å resolution, respectively. Each subunit of SCO4226 dimer adopts a canonical ferredoxin-like fold with five β-strands flanked by two α-helices. In the structure of Ni-SCO4226, four nickel ions are coordinated at the surface of the dimer. Further biochemical assays suggested that the binding of Ni2+ triggers the self-aggregation of SCO4226 in vitro. In addition, RT-qPCR assays demonstrated that the expression of SCO4226 gene in S. coelicolor is specifically up-regulated by the addition of Ni2+, but not other divalent ions such as Cu2+, Mn2+ or Co2+. All these results suggested that SCO4226 acts as a nickel binding protein, probably required for nickel sequestration and/or detoxification.

  17. 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.

  18. 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.

  19. The Wisconsin Rural Development Demonstration Project.

    ERIC Educational Resources Information Center

    McCoy, Palmer E.; Jones, Eleanor

    Nineteen Wisconsin counties were the site of the four-year Rural Development Demonstration Project, one of five such projects in the nation. Among the project's objectives were to accelerate sound economic, social, cultural and environmental development and to provide alternatives for living in non-metropolitan areas while providing greater…

  20. TOF-SIMS investigation of Streptomyces coelicolor, a mycelial bacterium

    NASA Astrophysics Data System (ADS)

    Vaidyanathan, Seetharaman; Fletcher, John S.; Lockyer, Nicholas P.; Vickerman, John C.

    2008-12-01

    Streptomyces coelicolor is a mycelial microorganism that produces several secondary metabolites, including antibiotics. The physiology of the organism has largely been investigated in liquid cultures due to ease of monitoring different physiological parameters and more homogeneous culture conditions. However, solid cultures reflect the natural physiology of the microorganism better, given that in its natural state it grows in the soil. Imaging mass spectrometry with TOF-SIMS and C 60+ primary ion beams offers a potential route to studying chemical changes at the molecular level, both intracellular and extracellular that can help in understanding the natural physiology of the microorganism. Here, we report the application of the technique for studying the lateral distribution of the chemical species detected in a population, grown in both liquid and solid cultures. The capability of the technique for studying biological systems with minimal system intervention is demonstrated.

  1. Developer/Demonstrator Directory. Second Edition.

    ERIC Educational Resources Information Center

    Far West Lab. for Educational Research and Development, San Francisco, CA.

    Many National Diffusion Network facilitators have expressed the desire for an expanded source book on Developer/Demonstrator projects that they can use in conjunction with "Educational Programs That Work" and the "Materials Inventory" in planning awareness and training activities and negotiating adoptions. This directory is a first step toward…

  2. 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.

  3. 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

  4. 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.

  5. The Level of AdpA Directly Affects Expression of Developmental Genes in Streptomyces coelicolor ▿ †

    PubMed Central

    Wolański, Marcin; Donczew, Rafał; Kois-Ostrowska, Agnieszka; Masiewicz, Paweł; Jakimowicz, Dagmara; Zakrzewska-Czerwińska, Jolanta

    2011-01-01

    AdpA is a key regulator of morphological differentiation in Streptomyces. In contrast to Streptomyces griseus, relatively little is known about AdpA protein functions in Streptomyces coelicolor. Here, we report for the first time the translation accumulation profile of the S. coelicolor adpA (adpASc) gene; the level of S. coelicolor AdpA (AdpASc) increased, reaching a maximum in the early stage of aerial mycelium formation (after 36 h), and remained relatively stable for the next several hours (48 to 60 h), and then the signal intensity decreased considerably. AdpASc specifically binds the adpASc promoter region in vitro and in vivo, suggesting that its expression is autoregulated; surprisingly, in contrast to S. griseus, the protein presumably acts as a transcriptional activator. We also demonstrate a direct influence of AdpASc on the expression of several genes whose products play key roles in the differentiation of S. coelicolor: STI, a protease inhibitor; RamR, an atypical response regulator that itself activates expression of the genes for a small modified peptide that is required for aerial growth; and ClpP1, an ATP-dependent protease. The diverse influence of AdpASc protein on the expression of the analyzed genes presumably results mainly from different affinities of AdpASc protein to individual promoters. PMID:21926228

  6. 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

  7. 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.

  8. 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.

  9. 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.

  10. Cardiolipin synthase is required for Streptomyces coelicolor morphogenesis

    PubMed Central

    Jyothikumar, Vinod; Klanbut, Khanungkan; Tiong, John; Roxburgh, James S.; Hunter, Iain S.; Smith, Terry K.; Herron, Paul R.

    2013-01-01

    Summary The fluid mosaic model has recently been amended to account for the existence of membrane domains enriched in certain phospholipids. In rod-shaped bacteria, the anionic phospholipid cardiolipin is enriched at the cell poles but its role in the morphogenesis of the filamentous bacterium Streptomyces coelicolor is unknown. It was impossible to delete clsA (cardiolipin synthase; SCO1389) unless complemented by a second copy of clsA elsewhere in the chromosome. When placed under the control of an inducible promoter, clsA expression, phospholipid profile and morphogenesis became inducer dependent. TLC analysis of phospholipid showed altered profiles upon depletion of clsA expression. Analysis of cardiolipin by mass spectrometry showed two distinct cardiolipin envelopes that reflected differences in acyl chain length; the level of the larger cardiolipin envelope was reduced in concert with clsA expression. ClsA-EGFP did not localize to specific locations, but cardiolipin itself showed enrichment at hyphal tips, branch points and anucleate regions. Quantitative analysis of hyphal dimensions showed that the mycelial architecture and the erection of aerial hyphae were affected by the expression of clsA. Overexpression of clsA resulted in weakened hyphal tips, misshaped aerial hyphae and anucleate spores and demonstrates that cardiolipin synthesis is a requirement for morphogenesis in Streptomyces. PMID:22409773

  11. 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.

  12. 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)

  13. 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...

  14. 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.

  15. 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. PMID:27648899

  16. 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.

  17. Identification and Characterization of FabA from the Type II Fatty Acid Synthase of Streptomyces coelicolor.

    PubMed

    Singh, Renu; Reynolds, Kevin A

    2016-01-22

    FabA is proposed to catalyze the dehydration step of chain elongation in fatty acid and undecylprodiginine biosynthesis in Streptomyces coelicolor. Analysis of the S. coelicolor genome has revealed a fabA gene (SCO4636-SCO4637, encoding a heterodimer 3-hydroxyacyl-ACP dehydratase). Herein, we report the identification and characterization of the corresponding gene products. Kinetic analysis has demonstrated that FabA is capable of utilizing various chain lengths of straight- and branched-chain 3-hydroxyacyl-NAC substrates. Additionally, FabA does not discriminate between acyl carrier proteins (ACPs) from primary and secondary metabolism. These data provide the first experimental evidence that FabA has 3-hydroxyacyl-ACP dehydratase activity and processes intermediates for both biosynthetic pathways.

  18. Tryptophan catabolism via kynurenine production in Streptomyces coelicolor: identification of three genes coding for the enzymes of tryptophan to anthranilate pathway.

    PubMed

    Zummo, F P; Marineo, S; Pace, A; Civiletti, F; Giardina, A; Puglia, A M

    2012-05-01

    Most enzymes involved in tryptophan catabolism via kynurenine formation are highly conserved in Prokaryotes and Eukaryotes. In humans, alterations of this pathway have been related to different pathologies mainly involving the central nervous system. In Bacteria, tryptophan and some of its derivates are important antibiotic precursors. Tryptophan degradation via kynurenine formation involves two different pathways: the eukaryotic kynurenine pathway, also recently found in some bacteria, and the tryptophan-to-anthranilate pathway, which is widespread in microorganisms. The latter produces anthranilate using three enzymes also involved in the kynurenine pathway: tryptophan 2,3-dioxygenase (TDO), kynureninase (KYN), and kynurenine formamidase (KFA). In Streptomyces coelicolor, where it had not been demonstrated which genes code for these enzymes, tryptophan seems to be important for the calcium- dependent antibiotic (CDA) production. In this study, we describe three adjacent genes of S. coelicolor (SCO3644, SCO3645, and SCO3646), demonstrating their involvement in the tryptophan-to-anthranilate pathway: SCO3644 codes for a KFA, SCO3645 for a KYN and SCO3646 for a TDO. Therefore, these genes can be considered as homologous respectively to kynB, kynU, and kynA of other microorganisms and belong to a constitutive catabolic pathway in S. coelicolor, which expression increases during the stationary phase of a culture grown in the presence of tryptophan. Moreover, the S. coelicolor ΔkynU strain, in which SCO3645 gene is deleted, produces higher amounts of CDA compared to the wild-type strain. Overall, these results describe a pathway, which is used by S. coelicolor to catabolize tryptophan and that could be inactivated to increase antibiotic production.

  19. 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.

  20. 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

  1. A versatile PCR-based tandem epitope tagging system for Streptomyces coelicolor genome.

    PubMed

    Kim, Ji-Nu; Yi, Jeong Sang; Lee, Bo-Rahm; Kim, Eun-Jung; Kim, Min Woo; Song, Yoseb; Cho, Byung-Kwan; Kim, Byung-Gee

    2012-07-20

    Epitope tagging approaches have been widely used for the analysis of functions, interactions and subcellular distributions of proteins. However, incorporating epitope sequence into protein loci in Streptomyces is time-consuming procedure due to the absence of the versatile tagging methods. Here, we developed a versatile PCR-based tandem epitope tagging tool for the Streptomyces genome engineering. We constructed a series of template plasmids that carry repeated sequence of c-myc epitope, Flp recombinase target (FRT) sites, and apramycin resistance marker to insert epitope tags into any desired spot of the chromosomal loci. A DNA module which includes the tandem epitope-encoding sequence and a selectable marker was amplified by PCR with primers that carry homologous extensions to the last portion and downstream region of the targeted gene. We fused the epitope tags at the 3' region of global transcription factors of Streptomyces coelicolor to test the validity of this system. The proper insertion of the epitope tag was confirmed by PCR and western blot analysis. The recombinants showed the identical phenotype to the wild-type that proved the conservation of in vivo function of the tagged proteins. Finally, the direct binding targets were successfully detected by chromatin immunoprecipitation with the increase in the signal-to-noise ratio. The epitope tagging system describes here would provide wide applications to study the protein functions in S. coelicolor. PMID:22704935

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

    PubMed

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

    2013-02-15

    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.

  3. 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

  4. 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.

  5. Allantoin catabolism influences the production of antibiotics in Streptomyces coelicolor.

    PubMed

    Navone, Laura; Casati, Paula; Licona-Cassani, Cuauhtémoc; Marcellin, Esteban; Nielsen, Lars K; Rodriguez, Eduardo; Gramajo, Hugo

    2014-01-01

    Purines are a primary source of carbon and nitrogen in soil; however, their metabolism is poorly understood in Streptomyces. Using a combination of proteomics, metabolomics, and metabolic engineering, we characterized the allantoin pathway in Streptomyces coelicolor. When cells grew in glucose minimal medium with allantoin as the sole nitrogen source, quantitative proteomics identified 38 enzymes upregulated and 28 downregulated. This allowed identifying six new functional enzymes involved in allantoin metabolism in S. coelicolor. From those, using a combination of biochemical and genetic engineering tools, it was found that allantoinase (EC 3.5.2.5) and allantoicase (EC 3.5.3.4) are essential for allantoin metabolism in S. coelicolor. Metabolomics showed that under these growth conditions, there is a significant intracellular accumulation of urea and amino acids, which eventually results in urea and ammonium release into the culture medium. Antibiotic production of a urease mutant strain showed that the catabolism of allantoin, and the subsequent release of ammonium, inhibits antibiotic production. These observations link the antibiotic production impairment with an imbalance in nitrogen metabolism and provide the first evidence of an interaction between purine metabolism and antibiotic biosynthesis.

  6. 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

  7. ArgR of Streptomyces coelicolor is a versatile regulator.

    PubMed

    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.

  8. 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...

  9. 10 CFR 1021.212 - Research, development, demonstration, and testing.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ..., development, demonstration, and testing for new technologies (40 CFR 1502.4(c)(3)). Adoption of such programs... development or restrict later alternatives, as discussed at 40 CFR 1502.4(c)(3). (c) For subsequent phases...

  10. 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.

  11. 17 CFR 256.188 - Research, development, or demonstration expenditures.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 17 Commodity and Securities Exchanges 3 2010-04-01 2010-04-01 false Research, development, or... COMPANIES, PUBLIC UTILITY HOLDING COMPANY ACT OF 1935 4. Deferred Debits § 256.188 Research, development, or... of all expenditures for research, development or demonstration undertaken by or sponsored through...

  12. 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)

  13. 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

  14. Development Status of the CECE Cryogenic Deep Throttling Demonstrator Engine

    NASA Technical Reports Server (NTRS)

    2008-01-01

    As one of the first technology development programs awarded by NASA under the U.S. Space Exploration Policy (USSEP), the Pratt & Whitney Rocketdyne (PWR) Deep Throttling, Common Extensible Cryogenic Engine (CECE) program was selected by NASA in November 2004 to begin technology development and demonstration toward a deep throttling, cryogenic engine supporting ongoing trade studies for NASA's Lunar Lander descent stage. The CECE program leverages the maturity and previous investment of a flight-proven hydrogen/oxygen expander cycle engine, the PWR RLI0, to develop and demonstrate an unprecedented combination of reliability, safety, durability, throttlability, and restart capabilities in a high-energy, cryogenic engine. The testbed selected for the deep throttling demonstration phases of this program was a minimally modified RL10 engine, allowing for maximum current production engine commonality and extensibility with minimum program cost. Two series of demonstrator engine tests, the first in April-May 2006 and the second in March-April 2007, have demonstrated in excess of 10:1 throttling of the hydrogen/oxygen expander cycle engine. Both test series have explored a combustion instability ("chug") environment at low throttled power levels. These tests have provided an early demonstration of an enabling cryogenic propulsion concept with invaluable system-level technology data acquisition toward design and development risk mitigation for future CECE Demonstrator engine tests.

  15. 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.

  16. Structure and function of the RedJ protein, a thioesterase from the prodiginine biosynthetic pathway in Streptomyces coelicolor.

    PubMed

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

    2011-06-24

    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.

  17. 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

  18. 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 evaluated... development or restrict later alternatives, as discussed at 40 CFR 1502.4(c)(3). (c) For subsequent phases...

  19. 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 evaluated... development or restrict later alternatives, as discussed at 40 CFR 1502.4(c)(3). (c) For subsequent phases...

  20. 10 CFR 1021.212 - Research, development, demonstration, and testing.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ..., 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 evaluated... development or restrict later alternatives, as discussed at 40 CFR 1502.4(c)(3). (c) For subsequent phases...

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

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 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 programs... development or restrict later alternatives, as discussed at 40 CFR 1502.4(c)(3). (c) For subsequent phases...

  2. Bioprocess intensification of antibiotic production by Streptomyces coelicolor A3(2) in micro-porous culture.

    PubMed

    Ndlovu, T M; Ward, A C; Glassey, J; Eskildsen, J; Akay, G

    2015-04-01

    A novel functionalized micro-porous matrix was developed with well-controlled physicochemical proprieties such as pore size and surface chemistry. The matrix was used as a solid support in the growth of "Streptomyces coelicolor" A3(2) to enhance the production of antibiotics. The results shown support a higher production of prodigiosin and actinorhodin with overall production increase of 2-5 and 6-17, respectively, compared to conventional submerged liquid culture, offering a potential improvement in volumetric productivity. Scanning Electron Microscopy was used to evaluate pore size as well as bacterial adhesion, penetration, proliferation and migration within the micro-porous matrix.

  3. Optimized submerged batch fermentation strategy for systems scale studies of metabolic switching in Streptomyces coelicolor A3(2)

    PubMed Central

    2012-01-01

    Background Systems biology approaches to study metabolic switching in Streptomyces coelicolor A3(2) depend on cultivation conditions ensuring high reproducibility and distinct phases of culture growth and secondary metabolite production. In addition, biomass concentrations must be sufficiently high to allow for extensive time-series sampling before occurrence of a given nutrient depletion for transition triggering. The present study describes for the first time the development of a dedicated optimized submerged batch fermentation strategy as the basis for highly time-resolved systems biology studies of metabolic switching in S. coelicolor A3(2). Results By a step-wise approach, cultivation conditions and two fully defined cultivation media were developed and evaluated using strain M145 of S. coelicolor A3(2), providing a high degree of cultivation reproducibility and enabling reliable studies of the effect of phosphate depletion and L-glutamate depletion on the metabolic transition to antibiotic production phase. Interestingly, both of the two carbon sources provided, D-glucose and L-glutamate, were found to be necessary in order to maintain high growth rates and prevent secondary metabolite production before nutrient depletion. Comparative analysis of batch cultivations with (i) both L-glutamate and D-glucose in excess, (ii) L-glutamate depletion and D-glucose in excess, (iii) L-glutamate as the sole source of carbon and (iv) D-glucose as the sole source of carbon, reveal a complex interplay of the two carbon sources in the bacterium's central carbon metabolism. Conclusions The present study presents for the first time a dedicated cultivation strategy fulfilling the requirements for systems biology studies of metabolic switching in S. coelicolor A3(2). Key results from labelling and cultivation experiments on either or both of the two carbon sources provided indicate that in the presence of D-glucose, L-glutamate was the preferred carbon source, while D

  4. 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.

  5. 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.

  6. Deciphering the Regulon of Streptomyces coelicolor AbrC3, a Positive Response Regulator of Antibiotic Production

    PubMed Central

    Rico, Sergio; Santamaría, Ramón I.; Yepes, Ana; Rodríguez, Héctor; Laing, Emma; Bucca, Giselda; Smith, Colin P.

    2014-01-01

    The atypical two-component system (TCS) AbrC1/C2/C3 (encoded by SCO4598, SCO4597, and SCO4596), comprising two histidine kinases (HKs) and a response regulator (RR), is crucial for antibiotic production in Streptomyces coelicolor and for morphological differentiation under certain nutritional conditions. In this study, we demonstrate that deletion of the RR-encoding gene, abrC3 (SCO4596), results in a dramatic decrease in actinorhodin (ACT) and undecylprodiginine (RED) production and delays morphological development. In contrast, the overexpression of abrC3 in the parent strain leads to a 33% increase in ACT production in liquid medium. Transcriptomic analysis and chromatin immunoprecipitation with microarray technology (ChIP-chip) analysis of the ΔabrC3 mutant and the parent strain revealed that AbrC3 directly controls ACT production by binding to the actII-ORF4 promoter region; this was independently verified by in vitro DNA-binding assays. This binding is dependent on the sequence 5′-GAASGSGRMS-3′. In contrast, the regulation of RED production is not due to direct binding of AbrC3 to either the redZ or redD promoter region. This study also revealed other members of the AbrC3 regulon: AbrC3 is a positive autoregulator which also binds to the promoter regions of SCO0736, bdtA (SCO3328), absR1 (SCO6992), and SCO6809. The direct targets share the 10-base consensus binding sequence and may be responsible for some of the phenotypes of the ΔabrC3 mutant. The identification of the AbrC3 regulon as part of the complex regulatory network governing antibiotic production widens our knowledge regarding TCS involvement in control of antibiotic synthesis and may contribute to the rational design of new hyperproducer host strains through genetic manipulation of such systems. PMID:24509929

  7. 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

  8. 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

  9. Genome sequence of a new Streptomyces coelicolor generalized transducing bacteriophage, ΦCAM.

    PubMed

    Monson, Rita; Salmond, George P C

    2012-12-01

    Streptomyces coelicolor is a model system for the study of Streptomyces, a genus of bacteria responsible for the production of many clinically important antibiotics. Here we report the genome sequence of ΦCAM, a new S. coelicolor generalized transducing bacteriophage, isolated from a soil sample originating from Lincolnshire, United Kingdom. Many open reading frames within ΦCAM shared high levels of similarity to a prophage from Salinispora tropica and a putative prophage in Streptomyces sp. strain C.

  10. Divalent transition-metal-ion stress induces prodigiosin biosynthesis in Streptomyces coelicolor M145: formation of coeligiosins.

    PubMed

    Morgenstern, Anne; Paetz, Christian; Behrend, Anne; Spiteller, Dieter

    2015-04-13

    The bacterium Streptomyces coelicolor M145 reacts to transition-metal-ion stress with myriad growth responses, leading to different phenotypes. In particular, in the presence of Co(2+) ions (0.7 mM) S. coelicolor consistently produced a red phenotype. This phenotype, when compared to the wild type, differed strongly in its production of volatile compounds as well as high molecular weight secondary metabolites. LC-MS analysis revealed that in the red phenotype the production of the prodigiosins, undecylprodigiosin and streptorubin B, was strongly induced and, in addition, several intense signals appeared in the LC-MS chromatogram. Using LC-MS/MS and NMR spectroscopy, two new prodigiosin derivatives were identified, that is, coeligiosin A and B, which contained an additional undecylpyrrolyl side chain attached to the central carbon of the tripyrrole ring system of undecylprodigiosin or streptorubin B. This example demonstrates that environmental factors such as heavy metal ion stress can not only induce the production of otherwise not observed metabolites from so called sleeping genes but alter the products from well-studied biosynthetic pathways.

  11. 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.

  12. Structural Insight into How Streptomyces coelicolor Maltosyl Transferase GlgE Binds α-Maltose 1-Phosphate and Forms a Maltosyl-enzyme Intermediate

    PubMed Central

    2014-01-01

    GlgE (EC 2.4.99.16) is an α-maltose 1-phosphate:(1→4)-α-d-glucan 4-α-d-maltosyltransferase of the CAZy glycoside hydrolase 13_3 family. It is the defining enzyme of a bacterial α-glucan biosynthetic pathway and is a genetically validated anti-tuberculosis target. It catalyzes the α-retaining transfer of maltosyl units from α-maltose 1-phosphate to maltooligosaccharides and is predicted to use a double-displacement mechanism. Evidence of this mechanism was obtained using a combination of site-directed mutagenesis of Streptomyces coelicolor GlgE isoform I, substrate analogues, protein crystallography, and mass spectrometry. The X-ray structures of α-maltose 1-phosphate bound to a D394A mutein and a β-2-deoxy-2-fluoromaltosyl-enzyme intermediate with a E423A mutein were determined. There are few examples of CAZy glycoside hydrolase family 13 members that have had their glycosyl-enzyme intermediate structures determined, and none before now have been obtained with a 2-deoxy-2-fluoro substrate analogue. The covalent modification of Asp394 was confirmed using mass spectrometry. A similar modification of wild-type GlgE proteins from S. coelicolor and Mycobacterium tuberculosis was also observed. Small-angle X-ray scattering of the M. tuberculosis enzyme revealed a homodimeric assembly similar to that of the S. coelicolor enzyme but with slightly differently oriented monomers. The deeper understanding of the structure–function relationships of S. coelicolor GlgE will aid the development of inhibitors of the M. tuberculosis enzyme. PMID:24689960

  13. 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

  14. 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.

  15. The development and demonstration of the metric assessment tool

    NASA Astrophysics Data System (ADS)

    Campbell, Cynthia A.; Gutterman, Gregory M.

    1993-09-01

    This study reflects the development and demonstration of the metric assessment tool. The purpose of the tool was to provide individuals the means to assess metrics and make improvements to the process measurement. The tool was developed using two critical attributes: customer satisfaction and process improvement. Once the tool was developed, a metric assessment process was designed to demonstrate the tool. Two metrics were selected from the Aeronautical Systems Center (ASC) and individuals with a working knowledge of the metric and process were selected for the demonstrations. Using a group support system at Armstrong Laboratory, Wright Patterson AFB, the group was asked to identify behaviors which might be motivated from the metric. Once the behaviors were identified, the group evaluated the behaviors against the critical attributes. From this assessment, behaviors were placed on the metric assessment tool. This tool clearly identified deficient behaviors and how they might distort the process measurement. From this information, the group was asked to generate improvement actions which would serve to eliminate or control deficient behaviors. With the elimination or control of deficient behaviors, the process measurement is improved and the organizational objective is better served.

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. 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

  1. 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.

  2. 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.

  3. 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

  4. Reactive Leadership: Divining, Developing, and Demonstrating Community Ontologies

    NASA Astrophysics Data System (ADS)

    Graybeal, J.

    2008-12-01

    The Marine Metadata Interoperability Project (known as MMI, on the web at http://marinemetadata.org) was formed to provide leadership in metadata practices to the marine science community. In 2004 this meant finding and writing about resources and best practices, which until then were all but invisible. In 2008 the scope is far wider, encompassing comprehensive guidance, collaborative community environments, and introduction and demonstration of advanced technologies to an increasingly interested scientific domain. MMI's technical leadership, based on experiences gained in the hydrologic community, emphasized the role ontologies could play in marine science. An early MMI workshop successfully incorporated a large number of community vocabularies, tools to harmonize them in a common ontological format, and the mapping of terms from vocabularies expressed in that format. That 2005 workshop demonstrated the connections to be made among different community vocabularies, and was well regarded by participants, but did not lead to widespread adoption of the tools, technologies, or even the vocabularies. Ontology development efforts for marine sensors and platforms showed intermittent progress, but again were not adopted or pushed toward completion. It is now 2008, and the marine community is increasingly attentive to a wide range of interoperability issues. A large part of the community has at least heard of "semantic interoperability", and many understand its critical role in finding and working with data. Demand for specific solutions, and for workable approaches, is becoming more vocal in the marine community. Yet there is still no encompassing model in place for achieving semantic interoperability, only simple operational registries have been set up for oceanographic community vocabularies, and only a few isolated applications demonstrate how semantic barriers can be overcome. Why has progress been so slow? Are good answers on the horizon? And if we build it, will the

  5. 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.

  6. Phosphorylation of ribosomal proteins influences subunit association and translation of poly (U) in Streptomyces coelicolor.

    PubMed

    Mikulík, Karel; Bobek, Jan; Ziková, Alice; Smětáková, Magdalena; Bezoušková, Silvie

    2011-03-01

    The occurrence of phosphorylated proteins in ribosomes of Streptomyces coelicolor was investigated. Little is known about which biological functions these posttranslational modifications might fulfil. A protein kinase associated with ribosomes phosphorylated six ribosomal proteins of the small subunit (S3, S4, S12, S13, S14 and S18) and seven ribosomal proteins of the large subunit (L2, L3, L7/L12, L16, L17, L23 and L27). The ribosomal proteins were phosphorylated mainly on the Ser/Thr residues. Phosphorylation of the ribosomal proteins influences ribosomal subunits association. Ribosomes with phosphorylated proteins were used to examine poly (U) translation activity. Phosphorylation induced about 50% decrease in polyphenylalanine synthesis. After preincubation of ribosomes with alkaline phosphatase the activity of ribosomes was greatly restored. Small differences were observed between phosphorylated and unphosphorylated ribosomes in the kinetic parameters of the binding of Phe-tRNA to the A-site of poly (U) programmed ribosomes, suggesting that the initial binding of Phe-tRNA is not significantly affected by phosphorylation. On contrary, the rate of peptidyl transferase was about two-fold lower than that in unphosphorylated ribosomes. The data presented demonstrate that phosphorylation of ribosomal proteins affects critical steps of protein synthesis.

  7. 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

  8. 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.

  9. Secretory production of an FAD cofactor-containing cytosolic enzyme (sorbitol-xylitol oxidase from Streptomyces coelicolor) using the twin-arginine translocation (Tat) pathway of Corynebacterium glutamicum.

    PubMed

    Scheele, Sandra; Oertel, Dan; Bongaerts, Johannes; Evers, Stefan; Hellmuth, Hendrik; Maurer, Karl-Heinz; Bott, Michael; Freudl, Roland

    2013-03-01

    Carbohydrate oxidases are biotechnologically interesting enzymes that require a tightly or covalently bound cofactor for activity. Using the industrial workhorse Corynebacterium glutamicum as the expression host, successful secretion of a normally cytosolic FAD cofactor-containing sorbitol-xylitol oxidase from Streptomyces coelicolor was achieved by using the twin-arginine translocation (Tat) protein export machinery for protein translocation across the cytoplasmic membrane. Our results demonstrate for the first time that, also for cofactor-containing proteins, a secretory production strategy is a feasible and promising alternative to conventional intracellular expression strategies.

  10. Secretory production of an FAD cofactor-containing cytosolic enzyme (sorbitol-xylitol oxidase from Streptomyces coelicolor) using the twin-arginine translocation (Tat) pathway of Corynebacterium glutamicum.

    PubMed

    Scheele, Sandra; Oertel, Dan; Bongaerts, Johannes; Evers, Stefan; Hellmuth, Hendrik; Maurer, Karl-Heinz; Bott, Michael; Freudl, Roland

    2013-03-01

    Carbohydrate oxidases are biotechnologically interesting enzymes that require a tightly or covalently bound cofactor for activity. Using the industrial workhorse Corynebacterium glutamicum as the expression host, successful secretion of a normally cytosolic FAD cofactor-containing sorbitol-xylitol oxidase from Streptomyces coelicolor was achieved by using the twin-arginine translocation (Tat) protein export machinery for protein translocation across the cytoplasmic membrane. Our results demonstrate for the first time that, also for cofactor-containing proteins, a secretory production strategy is a feasible and promising alternative to conventional intracellular expression strategies. PMID:23163932

  11. Regulation of myo-inositol catabolism by a GntR-type repressor SCO6974 in Streptomyces coelicolor.

    PubMed

    Yu, Lingjun; Li, Shuxian; Gao, Wenyan; Pan, Yuanyuan; Tan, Huarong; Liu, Gang

    2015-04-01

    Myo-inositol is important for Streptomyces growth and morphological differentiation. Genomic sequence analysis revealed a myo-inositol catabolic gene cluster in Streptomyces coelicolor. Disruption of the corresponding genes in this cluster abolished the bacterial growth on myo-inositol as a single carbon source. The transcriptions of these genes were remarkably enhanced by addition of myo-inositol in minimal medium. A putative regulatory gene SCO6974, encoding a GntR family protein, is situated in the cluster. Disruption of SCO6974 significantly enhanced the transcription of myo-inositol catabolic genes. SCO6974 was shown to interact with the promoter regions of myo-inositol catabolic genes using electrophoretic mobility shift assays. DNase I footprinting assays demonstrated that SCO6974 recognized a conserved palindromic sequence (A/T)TGT(A/C)N(G/T)(G/T)ACA(A/T). Base substitution of the conserved sequence completely abolished the binding of SCO6974 to the targets demonstrating that SCO6974 directly represses the transcriptions of myo-inositol catabolic genes. Furthermore, the disruption of SCO6974 was correlated with a reduced sporulation of S. coelicolor in mannitol soya flour medium and with the overproduction of actinorhodin and calcium-dependent antibiotic. The addition of myo-inositol suppressed the sporulation deficiency of the mutant, indicating that the effect could be related to a shortage in myo-inositol due to its enhanced catabolism in this strain. This enhanced myo-inositol catabolism likely yields dihydroxyacetone phosphate and acetyl-CoA that are indirect or direct precursors of the overproduced antibiotics.

  12. 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…

  13. Developing and Presenting Auditory Demonstrations: Two Sound Editor Programs

    ERIC Educational Resources Information Center

    Firment, Michael J.

    2006-01-01

    Experiencing examples of auditory phenomena can clarify textbook and lecture explanations. The addition of visual displays to auditory demonstrations can make them more understandable. Two sound editor programs, Audacity[R] and Adobe Audition Pro 2.0[R], provide excellent capabilities for the display and authoring of auditory demonstrations.…

  14. A jumping gene in streptomyces coelicolor A3(2).

    PubMed

    Sermonti, G; Lanfaloni, L; Micheli, M R

    1980-02-01

    The difficulty in mapping the gene for chloramphenicol resistance (cmlR) in Streptomyces coelicolor A3(2) stock strains is possibly due to its location on different sites of the chromosome in various mixed subclones. Fresh isolates from CmlR strains show single unequivocal locations of cmlR. The same holds for CmlR strains derived as revertants from CmlS variants. The two best established sites for cmlR are one between cysA and metA, the other at right of argA, possibly in the right empty arc of the map (Fig. 2). The cmlR gene was assumed to be on a transposon (SCTn1), together with a gene for arginine-succinate synthase (argG), a gene for chromosome transfer (tra) and a gene for aereal mycelium formation (amy). In a CmlR revertant, the cmlR gene appears disjoined from argG (Fig. 5), thus showing the ability of SCTnl to be split and partially transposed. The possible wide occurrence of transposons in the genus Streptomyces is discussed. PMID:6246399

  15. 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

  16. 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

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

    PubMed

    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

  18. 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

  19. Adventure Central: Applying the "Demonstration Plot" Concept to Youth Development

    ERIC Educational Resources Information Center

    Cochran, Graham; Arnett, Nate; Ferrari, Theresa M.

    2006-01-01

    Historically, land-grant university research stations focused on demonstrating successful farming methods. The land-grant mission and the principles at its foundation have broad applicability, and we believe the land-grant principles can be successfully applied in urban settings to a university's work with youth and families. In this article we…

  20. 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. PMID:21511036

  1. Pulsed Plasma Thruster Technology Development and Flight Demonstration Program

    NASA Technical Reports Server (NTRS)

    Peterson, Todd T.; Curran, Frank M.

    1998-01-01

    Because of anticipated near- and far-term mission needs for innovative new electric propulsion technologies, the NASA On-Board Propulsion program is sponsoring NASA Lewis Research Centers' Pulsed Plasma Thruster (PPT) technology development program to rapidly advance PPT technology. This is a joint effort of Lewis' On-Board Propulsion Branch and Space Flight Project Branch. Building on state-of-the-art technology developed and flown in the 1970's and 1980's for Department of Defense missions, newly developed PPT program designs are realizing significant gains in system mass reduction, thrust to mass ratio, total impulse to mass ratio, and variability of impulse bit.

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

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ...) SOLID WASTES CRITERIA FOR MUNICIPAL SOLID WASTE LANDFILLS General § 258.4 Research, development, and... landfills to assure protection of human health and the environment. Such permits shall: (1) Provide for...

  5. 40 CFR 258.4 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ...) SOLID WASTES CRITERIA FOR MUNICIPAL SOLID WASTE LANDFILLS General § 258.4 Research, development, and... landfills to assure protection of human health and the environment. Such permits shall: (1) Provide for...

  6. 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... landfills to assure protection of human health and the environment. Such permits shall: (1) Provide for...

  7. 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.

  8. 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.

  9. Proteomics-driven identification of SCO4677-dependent proteins in Streptomyces lividans and Streptomyces coelicolor.

    PubMed

    Choi, Si-Sun; Kim, Seon-Hye; Kim, Eung-Soo

    2010-03-01

    AfsR2 is a global regulatory protein which stimulates antibiotic biosynthesis in both Streptomyces lividans and S. coelicolor. Previously, various afsR2-dependent genes including a putative abaA-like regulatory gene, SCO4677, were identified through comparative DNA microarray analysis. To further identify the putative SCO4677-dependent proteins, the comparative proteomics-driven approach was applied to the SCO4677-overexpressing strains of S. lividans and S. coelicolor along with the wild-type strains. The 2D gel-electrophoresis gave approximately 277 protein spots for S. lividans and 207 protein spots for S. coelicolor, showing different protein expression patterns between the SCO4677-overexpressing strains and the wild-type strains. Further MALDI-TOF analysis revealed that only 18 proteins exhibited similar expression patterns in both S. lividans and S. coelicolor, suggesting that the SCO4677 could encode an abaA-like regulator which controls a few cross-species proteins as well as many species-specific proteins in Streptomyces species.

  10. Biosynthesis of the antibiotic actinorhodin. Analysis of blocked mutants of Streptomyces coelicolor.

    PubMed

    Cole, S P; Rudd, B A; Hopwood, D A; Chang, C J; Floss, H G

    1987-03-01

    From two types of class V act mutants of Streptomyces coelicolor two monomeric precursors of actinorhodin have been isolated and their structures determined. One is the known antibiotic kalafungin and the other a new compound. Their relationship to actinorhodin biosynthesis is discussed.

  11. 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.

  12. 40 CFR 258.4 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...) 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...

  13. 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...

  14. 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...

  15. RNA-Seq Analysis Reveals a Six-Gene SoxR Regulon in Streptomyces coelicolor

    PubMed Central

    Naseer, Nawar; Shapiro, Joshua A.; Chander, Monica

    2014-01-01

    The redox-regulated transcription factor SoxR is conserved in diverse bacteria, but emerging studies suggest that this protein plays distinct physiological roles in different bacteria. SoxR regulates a global oxidative stress response (involving >100 genes) against exogenous redox-cycling drugs in Escherichia coli and related enterics. In the antibiotic producers Streptomyces coelicolor and Pseudomonas aeruginosa, however, SoxR regulates a smaller number of genes that encode membrane transporters and proteins with homology to antibiotic-tailoring enzymes. In both S. coelicolor and P. aeruginosa, SoxR-regulated genes are expressed in stationary phase during the production of endogenously-produced redox-active antibiotics. These observations suggest that SoxR evolved to sense endogenous secondary metabolites and activate machinery to process and transport them in antibiotic-producing bacteria. Previous bioinformatics analysis that searched the genome for SoxR-binding sites in putative promoters defined a five-gene SoxR regulon in S. coelicolor including an ABC transporter, two oxidoreductases, a monooxygenase and an epimerase/dehydratase. Since this in silico screen may have missed potential SoxR-targets, we conducted a whole genome transcriptome comparison of wild type S. coelicolor and a soxR-deficient mutant in stationary phase using RNA-Seq. Our analysis revealed a sixth SoxR-regulated gene in S. coelicolor that encodes a putative quinone oxidoreductase. Knowledge of the full complement of genes regulated by SoxR will facilitate studies to elucidate the function of this regulatory molecule in antibiotic producers. PMID:25162599

  16. 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.

  17. 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

  18. Binary Cycle Geothermal Demonstration Power Plant New Developments

    SciTech Connect

    Lacy, Robert G.; Jacobson, William O.

    1980-12-01

    San Diego Gas and Electric Company (SDG and E) has been associated with geothermal exploration and development in the Imperial Valley since 1971. SDG and E currently has interests in the four geothermal reservoirs shown. Major SDG and E activities have included drilling and flow testing geothermal exploration wells, feasibility and process flow studies, small-scale field testing of power processes and equipment, and pilot plant scale test facility design, construction and operation. Supporting activities have included geothermal leasing, acquisition of land and water rights, pursual of a major new transmission line to carry Imperial Valley geothermal and other sources of power to San Diego, and support of Magma Electric's 10 MW East Mesa Geothermal Power Plant.

  19. 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

  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 Central

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

    2011-01-01

    Summary 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. PMID:21342469

  1. 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

  2. The role of absC, a novel regulatory gene for secondary metabolism, in zinc-dependent antibiotic production in Streptomyces coelicolor A3(2).

    PubMed

    Hesketh, Andy; Kock, Holger; Mootien, Saraspadee; Bibb, Mervyn

    2009-12-01

    The availability of zinc was shown to have a marked influence on the biosynthesis of actinorhodin in Streptomyces coelicolor A3(2). Production of actinorhodin and undecylprodigiosin was abolished when a novel pleiotropic regulatory gene, absC, was deleted, but only when zinc concentrations were low. AbsC was shown to control expression of the gene cluster encoding production of coelibactin, an uncharacterized non-ribosomally synthesized peptide with predicted siderophore-like activity, and the observed defect in antibiotic production was found to result from elevated expression of this gene cluster. Promoter regions in the coelibactin cluster contain predicted binding motifs for the zinc-responsive regulator Zur, and dual regulation of coelibactin expression by zur and absC was demonstrated using strains engineered to contain deletions in either or both of these genes. An AbsC binding site was identified in a divergent promoter region within the coelibactin biosynthetic gene cluster, adjacent to a putative Zur binding site. Repression of the coelibactin gene cluster by both AbsC and Zur appears to be required to maintain appropriate intracellular levels of zinc in S. coelicolor.

  3. Carbon-Flux Distribution within Streptomyces coelicolor Metabolism: A Comparison between the Actinorhodin-Producing Strain M145 and Its Non-Producing Derivative M1146

    PubMed Central

    Coze, Fabien; Gilard, Françoise; Tcherkez, Guillaume; Virolle, Marie-Joëlle; Guyonvarch, Armel

    2013-01-01

    Metabolic Flux Analysis is now viewed as essential to elucidate the metabolic pattern of cells and to design appropriate genetic engineering strategies to improve strain performance and production processes. Here, we investigated carbon flux distribution in two Streptomyces coelicolor A3 (2) strains: the wild type M145 and its derivative mutant M1146, in which gene clusters encoding the four main antibiotic biosynthetic pathways were deleted. Metabolic Flux Analysis and 13C-labeling allowed us to reconstruct a flux map under steady-state conditions for both strains. The mutant strain M1146 showed a higher growth rate, a higher flux through the pentose phosphate pathway and a higher flux through the anaplerotic phosphoenolpyruvate carboxylase. In that strain, glucose uptake and the flux through the Krebs cycle were lower than in M145. The enhanced flux through the pentose phosphate pathway in M1146 is thought to generate NADPH enough to face higher needs for biomass biosynthesis and other processes. In both strains, the production of NADPH was higher than NADPH needs, suggesting a key role for nicotinamide nucleotide transhydrogenase for redox homeostasis. ATP production is also likely to exceed metabolic ATP needs, indicating that ATP consumption for maintenance is substantial.Our results further suggest a possible competition between actinorhodin and triacylglycerol biosynthetic pathways for their common precursor, acetyl-CoA. These findings may be instrumental in developing new strategies exploiting S. coelicolor as a platform for the production of bio-based products of industrial interest. PMID:24376790

  4. Prioritizing orphan proteins for further study using phylogenomics and gene expression profiles in Streptomyces coelicolor

    PubMed Central

    2011-01-01

    Background Streptomyces coelicolor, a model organism of antibiotic producing bacteria, has one of the largest genomes of the bacterial kingdom, including 7825 predicted protein coding genes. A large number of these genes, nearly 34%, are functionally orphan (hypothetical proteins with unknown function). However, in gene expression time course data, many of these functionally orphan genes show interesting expression patterns. Results In this paper, we analyzed all functionally orphan genes of Streptomyces coelicolor and identified a list of "high priority" orphans by combining gene expression analysis and additional phylogenetic information (i.e. the level of evolutionary conservation of each protein). Conclusions The prioritized orphan genes are promising candidates to be examined experimentally in the lab for further characterization of their function. PMID:21899768

  5. Function and Evolution of Two Forms of SecDF Homologs in Streptomyces coelicolor

    PubMed Central

    Zhou, Zhan; Li, Yudong; Sun, Ning; Sun, Zhihao; Lv, Longxian; Wang, Yufeng; Shen, Libing; Li, Yong-Quan

    2014-01-01

    The general secretion (Sec) pathway plays a prominent role in bacterial protein export, and the accessory component SecDF has been shown to improve transportation efficiency. Inspection of Streptomyces coelicolor genome reveals the unexpected presence of two different forms of secDF homologous genes: one in fused form (secDF) and the other in separated form (secD and secF). However, the functional role of two SecDF homologs in S. coelicolor has not yet been determined. Transcriptional analysis of secDF homologs reveals that these genes are constitutively expressed. However, the transcript levels of secD and secF are much higher than that of secDF in S. coelicolor. Deletion of secDF or/and secD/secF in S. coelicolor did result in reduced secretion efficiency of Xylanase A and Amylase C, suggesting that they may have redundant functions for Sec-dependent translocation pathway. Moreover, our results also indicate that SecD/SecF plays a more prominent role than SecDF in protein translocation. Evolutionary analysis suggests that the fused and separated SecDF homologs in Streptomyces may have disparate evolutionary ancestries. SecD/SecF may be originated from vertical transmission of existing components from ancestor of Streptomyces species. However, SecDF may be derived from bacterial ancestors through horizontal gene transfer. Alternately, it is also plausible that SecDF may have arisen through additional gene duplication and fusion events. The acquisition of a second copy may confer a selective benefit to Streptomyces by enhancing protein transport capacity. Taken together, our results provide new insights into the potential biological function and evolutionary aspects of the prokaryotic SecDF complex. PMID:25140821

  6. 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

  7. 18 CFR 367.1880 - Account 188, Research, development, or demonstration expenditures.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 188, Research, development, or demonstration expenditures. (a) This account must be charged with the cost of all expenditures coming within the meaning of research, development and demonstration (RD&D) of..., development, and demonstration expenditures that are nonrecurring and that would distort the annual...

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

    PubMed

    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

  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. 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.

  11. Direct bio-electrocatalysis of O2 reduction by Streptomyces coelicolor laccase orientated at promoter-modified graphite electrodes.

    PubMed

    Lörcher, Samuel; Lopes, Paula; Kartashov, Andrey; Ferapontova, Elena E

    2013-07-22

    Bacterial laccase from Streptomyces coelicolor (SLAC) has been immobilised and orientated at promoter (pyrene and neocuproine)-modified electrodes productively both for direct electron transfer (ET) between the electrode and the T1 Cu site of SLAC and direct (unmediated) bio-electrocatalysis of dioxygen reduction. Its T1 Cu potential ranges between 471 and 318 mV versus the normal hydrogen electrode, at pH 5.5 and 8, respectively; this value is dependent both on the solution pH and electrode modification. In the presence of O2, Cu of the T2/T3 trinuclear centre is distinguished electrochemically at 748-623 mV. Depending on the promoter nature, different orientations of SLAC at pyrene- and neocuproine-modified electrodes can be followed from the kinetic analysis of the ET rates. Bio-electrocatalytic reduction of oxygen starts from the T1 Cu potentials of SLAC, and is most efficient at the promoter-modified electrodes, thereby demonstrating good performance both in neutral and basic media and in solutions with a high NaCl content, such as sea water. The obtained results allow consideration of a broader bioenergetic application of laccases as biocathodes operating directly in such environmental media as sea water and physiological fluids.

  12. A novel Streptomyces gene, samR, with different effects on differentiation of Streptomyces ansochromogenes and Streptomyces coelicolor.

    PubMed

    Tan, Huarong; Tian, Yuqing; Yang, Haihua; Liu, Gang; Nie, Liping

    2002-03-01

    A 1.4-kb DNA fragment from Streptomyces ansochromogenes accelerated mycelium formation of S. ansochromogenes when present on a multicopy plasmid. The DNA fragment contains one complete open reading frame, designated samR, encoding a protein with 213 amino acids that contains a likely DNA-binding helix-turn-helix motif close to its N-terminus. The deduced SamR protein resembles the product of the hppR gene, which is involved in the regulation of catabolism of 3-(3-hydroxyphenyl) propionate in Rhodococcus globerulus. A samR disruption mutant was constructed that presented a bald phenotype and failed to form aerial hyphae and spores. We suggest that samR plays an important role in the emergence of aerial hyphae from substrate mycelium. An almost identical gene of Streptomyces coelicolor was also subjected to gene disruption. Surprisingly, the mutant was able to develop an aerial mycelium, but it remained white and deficient in sporulation instead of forming gray spores. PMID:11907684

  13. Intracellular Metabolite Pool Changes in Response to Nutrient Depletion Induced Metabolic Switching in Streptomyces coelicolor.

    PubMed

    Wentzel, Alexander; Sletta, Havard; Ellingsen, Trond E; Bruheim, Per

    2012-02-17

    A metabolite profiling study of the antibiotic producing bacterium Streptomyces coelicolor A3(2) has been performed. The aim of this study was to monitor intracellular metabolite pool changes occurring as strains of S. coelicolor react to nutrient depletion with metabolic re-modeling, so-called metabolic switching, and transition from growth to secondary metabolite production phase. Two different culture media were applied, providing depletion of the key nutrients phosphate and L-glutamate, respectively, as the triggers for metabolic switching. Targeted GC-MS and LC-MS methods were employed to quantify important primary metabolite groups like amino acids, organic acids, sugar phosphates and other phosphorylated metabolites, and nucleotides in time-course samples withdrawn from fully-controlled batch fermentations. A general decline, starting already in the early growth phase, was observed for nucleotide pools and phosphorylated metabolite pools for both the phosphate and glutamate limited cultures. The change in amino acid and organic acid pools were more scattered, especially in the phosphate limited situation while a general decrease in amino acid and non-amino organic acid pools was observed in the L-glutamate limited situation. A phoP deletion mutant showed basically the same metabolite pool changes as the wild-type strain M145 when cultivated on phosphate limited medium. This implies that the inactivation of the phoP gene has only little effect on the detected metabolite levels in the cell. The energy charge was found to be relatively constant during growth, transition and secondary metabolite production phase. The results of this study and the employed targeted metabolite profiling methodology are directly relevant for the evaluation of precursor metabolite and energy supply for both natural and heterologous production of secondary metabolites in S. coelicolor.

  14. Intracellular Metabolite Pool Changes in Response to Nutrient Depletion Induced Metabolic Switching in Streptomyces coelicolor

    PubMed Central

    Wentzel, Alexander; Sletta, Havard; Consortium, Stream; Ellingsen, Trond E.; Bruheim, Per

    2012-01-01

    A metabolite profiling study of the antibiotic producing bacterium Streptomyces coelicolor A3(2) has been performed. The aim of this study was to monitor intracellular metabolite pool changes occurring as strains of S. coelicolor react to nutrient depletion with metabolic re-modeling, so-called metabolic switching, and transition from growth to secondary metabolite production phase. Two different culture media were applied, providing depletion of the key nutrients phosphate and L-glutamate, respectively, as the triggers for metabolic switching. Targeted GC-MS and LC-MS methods were employed to quantify important primary metabolite groups like amino acids, organic acids, sugar phosphates and other phosphorylated metabolites, and nucleotides in time-course samples withdrawn from fully-controlled batch fermentations. A general decline, starting already in the early growth phase, was observed for nucleotide pools and phosphorylated metabolite pools for both the phosphate and glutamate limited cultures. The change in amino acid and organic acid pools were more scattered, especially in the phosphate limited situation while a general decrease in amino acid and non-amino organic acid pools was observed in the L-glutamate limited situation. A phoP deletion mutant showed basically the same metabolite pool changes as the wild-type strain M145 when cultivated on phosphate limited medium. This implies that the inactivation of the phoP gene has only little effect on the detected metabolite levels in the cell. The energy charge was found to be relatively constant during growth, transition and secondary metabolite production phase. The results of this study and the employed targeted metabolite profiling methodology are directly relevant for the evaluation of precursor metabolite and energy supply for both natural and heterologous production of secondary metabolites in S. coelicolor. PMID:24957373

  15. Genetics of the phage growth limitation (Pgl) system of Streptomyces coelicolor A3(2).

    PubMed

    Sumby, Paul; Smith, Margaret C M

    2002-04-01

    The phage growth limitation (Pgl) system, encoded by Streptomyces coelicolor A3(2), confers protection against the temperate bacteriophage phiC31 and its homoimmune relatives. The Pgl phenotype is characterized by the ability of Pgl+ hosts to support a phage burst on initial infection but subsequent cycles are severely attenuated. Previously, two adjacent genes pglY and pglZ were shown to be required for Pgl. It had been shown by Southern blotting that Streptomyces lividans, a close relative of S. coelicolor and naturally Pgl-, does not contain homologues of pglYZ and that introduction of pglYZ into S. lividans is not sufficient to confer a Pgl+ phenotype. Moreover, the mechanism of the Pgl+<--> Pgl- phase variation associated with this phenotype is also not understood. Here we describe two novel genes, pglW and pglX, that were shown to be part of this system by complementation of Pgl- mutants and by insertional mutagenesis. pglW encodes a 169 kDa protein that includes putative motifs for both serine/threonine protein kinase activity and DNA binding. pglX encodes a 136 kDa protein with putative adenine-specific DNA methyltransferase activity. pglW and pglX have overlapping stop-start codons suggesting transcriptional and translational coupling. S1 mapping of transcripts initiating up-stream of pglW indicated that, like pglYZ, pglWX is expressed in uninfected cultures. A homologue of pglX with 76% amino acid identity was identified in S. coelicolor, and insertional mutagenesis indicated that this gene was not required for the Pgl+ phenotype. Southern blots indicated that S. lividans does not contain homologues of pglW or pglX. A plasmid encoding pglWXYZ was able to confer the Pgl+ phenotype to S. lividans implying that these four genes constitute the whole system.

  16. AfsR as an integrator of signals that are sensed by multiple serine/threonine kinases in Streptomyces coelicolor A3(2).

    PubMed

    Horinouchi, Sueharu

    2003-08-01

    The genome sequence of Streptomyces coelicolor A3(2) has revealed the presence of about 40 protein serine/threonine or tyrosine kinases. AfsK, which is able to phosphorylate AfsR, a transcriptional activator with ATPase activity, represents the first instance in which a bacterial Hanks-type protein kinase phosphorylates a specific protein and exerts biologically important functions. The AfsK-AfsR system in S. coelicolor A3(2) globally controls secondary metabolism. The signal transduction pathway so far demonstrated or suggested is as follows: AfsK loosely attached to the membrane autophosphorylates threonine and serine residues, perhaps on sensing some external stimulus, and enhances its kinase activity. The kinase activity is modulated by KbpA, an AfsK-binding protein, by means of protein-protein interactions. The activated AfsK phosphorylates threonine and serine residues of AfsR in the cytoplasm, by which the DNA-binding activity of AfsR is greatly enhanced. In addition to AfsK, other kinases-including PkaG and AfsL-also phosphorylate AfsR, suggesting that AfsR serves as an integrator of multiple signals sensed by these kinases. The phosphorylated AfsR binds the promoter of afsS, which encodes a protein of 63 amino acids, and forms a closed complex with RNA polymerase. The closed complex is then converted to a transcriptionally active open complex by the energy available from ATP hydrolysis by AfsR. AfsS induced in this way activates transcription of pathway-specific transcriptional activators, such as actII-ORF4 for actinorhodin production and redD for undecylprodigiosin, in an as yet unknown manner.

  17. 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

  18. Identification of new developmentally regulated genes involved in Streptomyces coelicolor sporulation

    PubMed Central

    2013-01-01

    Background The sporulation of aerial hyphae of Streptomyces coelicolor is a complex developmental process. Only a limited number of the genes involved in this intriguing morphological differentiation programme are known, including some key regulatory genes. The aim of this study was to expand our knowledge of the gene repertoire involved in S. coelicolor sporulation. Results We report a DNA microarray-based investigation of developmentally controlled gene expression in S. coelicolor. By comparing global transcription patterns of the wild-type parent and two mutants lacking key regulators of aerial hyphal sporulation, we found a total of 114 genes that had significantly different expression in at least one of the two mutants compared to the wild-type during sporulation. A whiA mutant showed the largest effects on gene expression, while only a few genes were specifically affected by whiH mutation. Seven new sporulation loci were investigated in more detail with respect to expression patterns and mutant phenotypes. These included SCO7449-7451 that affect spore pigment biogenesis; SCO1773-1774 that encode an L-alanine dehydrogenase and a regulator-like protein and are required for maturation of spores; SCO3857 that encodes a protein highly similar to a nosiheptide resistance regulator and affects spore maturation; and four additional loci (SCO4421, SCO4157, SCO0934, SCO1195) that show developmental regulation but no overt mutant phenotype. Furthermore, we describe a new promoter-probe vector that takes advantage of the red fluorescent protein mCherry as a reporter of cell type-specific promoter activity. Conclusion Aerial hyphal sporulation in S. coelicolor is a technically challenging process for global transcriptomic investigations since it occurs only as a small fraction of the colony biomass and is not highly synchronized. Here we show that by comparing a wild-type to mutants lacking regulators that are specifically affecting processes in aerial hypha, it is

  19. 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

  20. 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.

  1. 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.

  2. Identification of the antibiotic determined by the SCP1 plasmid of Streptomyces coelicolor A3(2).

    PubMed

    Wright, L F; Hopwood, D A

    1976-07-01

    The antibiotic whose biosynthesis is determined by the SCP1 plasmid of Streptomyces coelicolor A3(2) has been characterized as the recently described methylenomycin A (2-methylene-cyclopentan-3-one-4,5-epoxy-4,5-dimethyl-1-carboxylic acid).

  3. Physiological role of germicidins in spore germination and hyphal elongation in Streptomyces coelicolor A3(2).

    PubMed

    Aoki, Yuu; Matsumoto, Daisuke; Kawaide, Hiroshi; Natsume, Masahiro

    2011-09-01

    Four germicidin homologs were isolated from a liquid culture of Streptomyces coelicolor A3(2). These were identified as germicidins A, B and C, and surugapyrone A (germicidin D). Absolute stereochemistry of the chiral center in germicidins A and C is determined to be S. All germicidins inhibited germination of S. coelicolor A3(2) spores above 1 μg ml(-1). S. coelicolor A3(2) spores collected from a single petri dish (9 cm i.d.) contained 5.4 μg of germicidin A (-2.7 × 10(-14) g per spore), which accounts for 2.3% of the spore extract, and contents of germicidins B, C and D were 0.2-0.8 μg. The activity of the spore extract corresponded well with the sum of the activity of each germicidin, which was estimated from the content and dose-response curve, which indicates that germicidins functions as self-germination inhibitors in S. coelicolor A3(2). Inhibitory action of germicidin A on spore germination was reversible and germicidin A inhibited not only spore germination but also hyphal elongation.

  4. 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.

  5. A plan for the development and demonstration of optical communications for deep space

    NASA Technical Reports Server (NTRS)

    Lesh, James R.; Deutsch, Leslie J.; Weber, William J.

    1991-01-01

    A development history, current status, and prospective development account are given for deep-space optical communications, giving attention to overall systems-related activities and to the generic technology developments required for their implementation. An account is given of the progression envisioned from laboratory subsystems demonstrations to the full-scale flight experiment for the Mars Communications Relay Orbiter missions.

  6. Ergothioneine protects Streptomyces coelicolor A3(2) from oxidative stresses.

    PubMed

    Nakajima, Shunsuke; Satoh, Yasuharu; Yanashima, Kentaro; Matsui, Tomomi; Dairi, Tohru

    2015-09-01

    Thiol compounds with low-molecular weight, such as glutathione, mycothiol (MSH), bacillithiol, and ergothioneine (ERG), are known to protect microorganisms from oxidative stresses. Mycobacteria and actinobacteria utilize both MSH and ERG. The biological functions of MSH in mycobacteria have been extensively studied by genetic and biochemical studies, which have suggested it has critical roles for detoxification in cells. In contrast, the biological functions of ERG remain ambiguous because its biosynthetic genes were only recently identified in Mycobacterium avium. In this study, we constructed mutants of Streptomyces coelicolor A3(2), in which either the MSH or ERG biosynthetic gene was disrupted, and examined their phenotypes. A mshC (SCO1663)-disruptant completely lost MSH productivity. In contrast, a disruptant of the egtA gene (SCO0910) encoding γ-glutamyl-cysteine synthetase unexpectedly retained reduced productivity of ERG, probably because of the use of l-cysteine instead of γ-glutamyl-cysteine. Both disruptants showed delayed growth at the late logarithmic phase and were more susceptible to hydrogen peroxide and cumene hydroperoxide than the parental strain. Interestingly, the ERG-disruptant, which still kept reduced ERG productivity, was more susceptible. Furthermore, the ERG-disruptant accumulated 5-fold more MSH than the parental strain. In contrast, the amount of ERG was almost the same between the MSH-disruptant and the parental strain. Taken together, our results suggest that ERG is more important than MSH in S. coelicolor A3(2). PMID:25683449

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

    PubMed

    Gomez-Escribano, Juan Pablo; Bibb, Mervyn J

    2011-03-01

    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.

  8. The Streptomyces coelicolor developmental transcription factor sigmaBldN is synthesized as a proprotein.

    PubMed

    Bibb, Maureen J; Buttner, Mark J

    2003-04-01

    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 sigma factors and that translation from the most strongly predicted start codon (GTG(1)) would give rise to a sigma 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 GTG(1) and that the primary translation product is a proprotein (pro-sigma(BldN)) that is proteolytically processed to a mature species (sigma(BldN)) 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-sigma(BldN) and the subsequent appearance of mature sigma(BldN), which was concomitant with aerial mycelium formation and the disappearance of pro-sigma(BldN). 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-sigma(BldN) processing.

  9. 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…

  10. Evaluating Child and Family Demonstration Initiatives: Lessons from the Comprehensive Child Development Program.

    ERIC Educational Resources Information Center

    Gilliam, Walter S.; Ripple, Carol H.; Zigler, Edward F.; Leiter, Valerie

    2000-01-01

    Argues that shortcomings in program implementation and evaluation contributed to failure to demonstrate effectiveness of the Comprehensive Child Development Program (CCDP). Articulates lessons from the CCDP, including addressing roles of demonstration projects and their evaluations, problems in evaluation early in program implementation, the…

  11. 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.

  12. 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.

  13. The two kinases, AbrC1 and AbrC2, of the atypical two-component system AbrC are needed to regulate antibiotic production and differentiation in Streptomyces coelicolor

    PubMed Central

    Rodríguez, Héctor; Rico, Sergio; Yepes, Ana; Franco-Echevarría, Elsa; Antoraz, Sergio; Santamaría, Ramón I.; Díaz, Margarita

    2015-01-01

    Two-component systems (TCSs) are the most important sensing mechanisms in bacteria. In Streptomyces, TCSs-mediated responses to environmental stimuli are involved in the regulation of antibiotic production. This study examines the individual role of two histidine kinases (HKs), AbrC1 and AbrC2, which form part of an atypical TCS in Streptomyces coelicolor. qRT-PCR analysis of the expression of both kinases demonstrated that both are expressed at similar levels in NB and NMMP media. Single deletion of abrC1 elicited a significant increase in antibiotic production, while deletion of abrC2 did not have any clear effect. The origin of this phenotype, probably related to the differential phosphorylation ability of the two kinases, was also explored indirectly, analyzing the toxic phenotypes associated with high levels of phosphorylated RR. The higher the AbrC3 regulator phosphorylation rate, the greater the cell toxicity. For the first time, the present work shows in Streptomyces the combined involvement of two different HKs in the response of a regulator to environmental signals. Regarding the possible applications of this research, the fact that an abrC1 deletion mutant overproduces three of the S. coelicolor antibiotics makes this strain an excellent candidate as a host for the heterologous production of secondary metabolites. PMID:26029189

  14. Integrated gasification combined-cycle research development and demonstration activities in the U.S.

    SciTech Connect

    Ness, H.M.

    1994-12-31

    The United States Department of Energy (DOE) has selected seven integrated gasification combined-cycle (IGCC) advanced power systems for demonstration in the Clean Coal Technology (CCT) Program. DOE`s Office of Fossil Energy, Morgantown Energy Technology Center, is managing a research development and demonstration (RD&D)program that supports the CCT program, and addresses long-term improvements in support of IGCC technology. This overview briefly describes the CCT projects and the supporting RD&D activities.

  15. 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.

  16. 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

  17. 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.

  18. Phase variation in the phage growth limitation system of Streptomyces coelicolor A3(2).

    PubMed

    Sumby, Paul; Smith, Margaret C M

    2003-08-01

    The phase-variable phage growth limitation (Pgl) system of Streptomyces coelicolor A3(2) is an unusual bacteriophage resistance mechanism that confers protection against the temperate phage phiC31 and homoimmune relatives. Pgl is subject to phase variation, and data presented here show that this is at least partially due to expansion and contraction of a polyguanine tract present within the putative adenine-specific DNA methyltransferase gene, pglX. Furthermore, the pglX paralogue SC6G9.02, here renamed pglS, was shown to be able to interfere with the Pgl phenotype, suggesting that PglS could provide an alternative activity to that conferred by PglX.

  19. 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

  20. 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.

  1. Managing Risk on a Technology Development Project/Advanced Mirror System Demonstrator

    NASA Technical Reports Server (NTRS)

    Byberg, Alicia; Russell, J. Kevin; Stahl, Phil (Technical Monitor)

    2002-01-01

    The risk management study applied to the Advanced Mirror System Demonstrator (AMSD), a precursor mirror technology development for the Next Generation Space Telescope (NGST) is documented. The AMSD will be developed as a segment of a lightweight primary mirror system that can be produced at a low cost and with a short manufacturing schedule. The technology gained from the program will support the risk mitigation strategy for the NGST, as well as other government agency space mirror programs.

  2. Genome-scale analysis of Streptomyces coelicolor A3(2) metabolism

    PubMed Central

    Borodina, Irina; Krabben, Preben; Nielsen, Jens

    2005-01-01

    Streptomyces are filamentous soil bacteria that produce more than half of the known microbial antibiotics. We present the first genome-scale metabolic model of a representative of this group—Streptomyces coelicolor A3(2). The metabolism reconstruction was based on annotated genes, physiological and biochemical information. The stoichiometric model includes 819 biochemical conversions and 152 transport reactions, accounting for a total of 971 reactions. Of the reactions in the network, 700 are unique, while the rest are iso-reactions. The network comprises 500 metabolites. A total of 711 open reading frames (ORFs) were included in the model, which corresponds to 13% of the ORFs with assigned function in the S. coelicolor A3(2) genome. In a comparative analysis with the Streptomyces avermitilis genome, we showed that the metabolic genes are highly conserved between these species and therefore the model is suitable for use with other Streptomycetes. Flux balance analysis was applied for studies of the reconstructed metabolic network and to assess its metabolic capabilities for growth and polyketides production. The model predictions of wild-type and mutants' growth on different carbon and nitrogen sources agreed with the experimental data in most cases. We estimated the impact of each reaction knockout on the growth of the in silico strain on 62 carbon sources and two nitrogen sources, thereby identifying the “core” of the essential reactions. We also illustrated how reconstruction of a metabolic network at the genome level can be used to fill gaps in genome annotation. PMID:15930493

  3. 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.

  4. Evaluation of the Stallings Classroom Management Staff Development Demonstration Project in Putnam County, West Virginia.

    ERIC Educational Resources Information Center

    Meehan, Merrill L.

    An evaluation is presented of the Stallings Classroom Management Staff Development Demonstration Project, designed to increase student achievement in basic skills through the use of research-based, systematic change in teachers' classroom management and organizational techniques. One objective of the evaluation was to utilize techniques and to…

  5. Cost Analysis of Air Force On-the-Job Training: Development and Demonstration of a Methodology.

    ERIC Educational Resources Information Center

    Eisele, Charles R.; And Others

    A research project was developed to construct and demonstrate a methodology for estimating the costs of conducting on-the-job training (OJT) in the Air Force. The project focused on the formal upgrade training to the three, five, and seven skill levels. Project efforts involved five major tasks: literature review, cost factor identification, cost…

  6. 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.

  7. Development of highly reliable advanced grid structure (HRAGS) demonstrator using FBG sensors

    NASA Astrophysics Data System (ADS)

    Takeya, H.; Sekine, K.; Kume, M.; Ozaki, T.; Takeda, N.; Tajima, N.

    2008-03-01

    There is a growing demand in recent years for lightweight structures in aircraft systems from the viewpoints of energy and cost savings. The authors have continued development of the Highly Reliable Advanced Grid Structure (HRAGS) for aircraft structure. HRAGS is provided with health monitoring functions that make use of Fiber Bragg Grating (FBG) sensors in advanced grid structures. To apply HRAGS technology to aircraft structures, a full-scale demonstrator visualizing the actual aircraft structure needs to be built and evaluated so that the effectiveness of the technology can be validated. So the authors selected the wing tip as the candidate structural member and proceeded to design and build a demonstrator. A box-structure was adopted as the structure for the wing-tip demonstrator, and HRAGS panels were used as the skin panels on the upper and lower surfaces of the structure. The demonstrator was designed using about 600 FBG sensors using a panel size of 1 x 2 m. By using the demonstrator, damage detection functions of HRAGS system were verified analytically. The results of the design and evaluation of the demonstrator are reported here.

  8. 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.

  9. Waste-heat research, development, demonstration and commercialization plan: Rankine-cycle bottoming systems. Executive summary

    SciTech Connect

    Not Available

    1980-01-01

    Organic and binary Rankine cycle (ORC) technology as potentially broad applications in recovering and converting waste heat to the useful energy form of electricity. ORC systems are particularly suited for recovering medium-grade exhaust heat (200 to 1000/sup 0/F), a form of waste energy released primarily in the generation of electricity and in industrial processes. Therefore, a Waste Heat Research, Development, Demonstration and Commercialization Plan (RDD and C Plan) has been formulated. The objective of the plan is to achieve significant market penetration of ORC technology by 1985. To accomplish this commercialization objective, the plan is structured around three key strategic elements: demonstration of technically and economically attractive ORC systems that meet the specific needs of commercial waste heat markets; stimulation of the demand for ORC products in these markets; and promotion of the development of a competitive industry to serve ORC markets efficiently. The development of this plan and ERDA's role in its implementation are discussed.

  10. 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.

  11. Characterization of FabG and FabI of the Streptomyces coelicolor dissociated fatty acid synthase.

    PubMed

    Singh, Renu; Reynolds, Kevin A

    2015-03-01

    Streptomyces coelicolor produces fatty acids for both primary metabolism and for biosynthesis of the secondary metabolite undecylprodiginine. The first and last reductive steps during the chain elongation cycle of fatty acid biosynthesis are catalyzed by FabG and FabI. The S. coelicolor genome sequence has one fabI gene (SCO1814) and three likely fabG genes (SCO1815, SCO1345, and SCO1846). We report the expression, purification, and characterization of the corresponding gene products. Kinetic analyses revealed that all three FabGs and FabI are capable of utilizing both straight and branched-chain β-ketoacyl-NAC and enoyl-NAC substrates, respectively. Furthermore, only SCO1345 differentiates between ACPs from both biosynthetic pathways. The data presented provide the first experimental evidence that SCO1815, SCO1346, and SCO1814 have the catalytic capability to process intermediates in both fatty acid and undecylprodiginine biosynthesis. PMID:25662938

  12. In vivo DNase I sensitivity of the Streptomyces coelicolor chromosome correlates with gene expression: implications for bacterial chromosome structure.

    PubMed

    McArthur, Michael; Bibb, Mervyn

    2006-01-01

    For a bacterium, Streptomyces coelicolor A3(2) contains a relatively large genome (8.7 Mb) with a complex and adaptive pattern of gene regulation. We discovered a correlation between the physical structure of the S.coelicolor genome and the transcriptional activity of the genes therein. Twelve genes were surveyed throughout 72 h of growth for both in vivo sensitivity to DNase I digestion and levels of transcription. DNase I-sensitivity correlated positively with transcript levels, implying that it was predictive of gene expression, and indicating increased accessibility of transcribed DNA. The genome was fractionated based on the sensitivity to DNase I digestion, with the low molecular weight (frequently cut) fraction highly enriched for actively transcribed sequences when compared to the infrequently cut fraction, which was representative of the entire genome. This approach will allow comparison of nucleoid proteins, and any modifications thereof, associated with transcriptionally active and inactive regions of the bacterial genome.

  13. Development of a 15 T $Nb_3Sn$ Accelerator Dipole Demonstrator at Fermilab

    SciTech Connect

    Novitski, I.; Andreev, N.; Barzi, E.; Carmichael, J.; Kashikhin, V. V.; Turrion, D.; Yu, M.; Zlobin, A. V.

    2015-01-01

    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 $Nb_3Sn$ 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.

  14. Development of a 15 T $Nb_3Sn$ Accelerator Dipole Demonstrator at Fermilab

    SciTech Connect

    Novitski, I.; Andreev, N.; Barzi, E.; Carmichael, J.; Kashikhin, V. V.; Turrion, D.; Yu, M.; Zlobin, A. V.

    2016-01-01

    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 $Nb_3Sn$ 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.

  15. 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

  16. 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

  17. 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

  18. 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.

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

    PubMed Central

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

    2012-01-01

    Summary 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 down-stream genes in the S. coelicolor transcriptional cascade. In Amycolatopsis sp. AA4, siderophore genes were down-regulated 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. PMID:22931126

  20. The role of IGCC in USDOE clean coal research, development and demonstration

    SciTech Connect

    Schmidt, D.K.; Rath, L.K.

    1993-09-01

    For many years, the US Department of Energy (DOE) has been funding research, development, and demonstration (RD&D) projects to develop advanced power generation technologies. The goal of this RD&D is private sector commercialization of technologies that will provide reasonably priced electricity and still meet stringent environmental standards. Integrated Gasification Combined-Cycle (IGCC) systems are emerging as one of the more attractive candidate technologies to meet this goal. The Morgantown Energy Technology Center (METC) has been assigned the responsibility for implementing IGCC projects in DOE`s Clean Coal RD&D program. Projects related to IGCC are briefly described.

  1. 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.

  2. 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.

  3. 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

  4. Development and demonstration of a lignite-pelletizing process. Phase II report

    SciTech Connect

    Not Available

    1980-01-01

    The current work began with scale-up of laboratory equipment to commercial size equipment. For this purpose, BCI used an existing pilot plant that had been assembled to pelletize and indurate taconite ore. BCI determined therewith that lignite pellets can be produced continuously on a pilot scale using the basic process developed in the laboratory. The resulting pellets were found to be similar to the laboratory pellets at equivalent binder compositions. Tests of product made during a 5-ton test run are reported. A 50-ton demonstration test run was made with the pilot plant. Pellet production was sustained for a two-week period. The lignite pelletizing process has, therefore, been developed to the point of demonstration in a 50-ton test. BCI has completed and cost estimated a conceptually designed 4000 TPD facility. BCI believes it has demonstrated a technically feasible process to agglomerate lignite by using an asphalt emulsion binder. Product quality is promising. Capital and operating costs appear acceptable to justify continuing support and development. The next step should focus on three areas: production development, process refinement, and cost reduction. For further development, BCI recommends consideration of a 5 to 10 ton/h pilot plant or a 20 to 40 ton/h module of a full sized plant, the lower first cost of the former being offset by the ability to incorporate the latter into a future production unit. Other specific recommendations are made for future study that could lead to process and cost improvements: Binder Formulation, disc Sizing, Drier Bed Depth, and Mixing Approach. Pellet use other than power plant fuel is considered.

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

    NASA Technical Reports Server (NTRS)

    Young, Roy M.; Montgomery, Edward E.; Montgomery, Sandy; 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.

  6. 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.

  7. 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.

  8. 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

  9. 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).

  10. 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.

  11. 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).

  12. 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.

  13. 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.

  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. 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.

  16. Development and demonstration of treatment technologies for the processing of US Department of Energy mixed waste

    SciTech Connect

    Berry, J.B.; Bloom, G.A.; Kuchynka, D.J.

    1994-06-01

    Mixed waste is defined as waste contaminated with chemically hazardous (governed by the Resource Conservation and Recovery Act) and radioactive species [governed by US Department of Energy (DOE) orders]. The Mixed Waste Integrated Program (MWIP) is responding to the need for DOE mixed waste treatment technologies that meet these dual regulatory requirements. MWIP is developing emerging and innovative treatment technologies to determine process feasibility. Technology demonstrations will be used to determine whether processes are superior to existing technologies in reducing risk, minimizing life-cycle cost, and improving process performance. The Program also provides a forum for stakeholder and customer involvement in the technology development process. MWIP is composed of six technical areas that support a mixed-waste treatment system: (1) systems analysis, (2) materials handling, (3) chemical/physical separation, (4) waste destruction and stabilization, (5) off-gas treatment, and (6) final waste form stabilization. The status of the technical initiatives and the current research, development, and demonstration in each of these areas is described in this paper.

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. 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. PMID:27572512

  2. 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.

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

    DOE PAGESBeta

    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 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

  4. Development of a 15 T $Nb_3Sn$ Accelerator Dipole Demonstrator at Fermilab

    DOE PAGESBeta

    Novitski, I.; Andreev, N.; Barzi, E.; Carmichael, J.; Kashikhin, V. V.; Turrion, D.; Yu, M.; Zlobin, A. V.

    2016-01-01

    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 $Nb_3Sn$ 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. Themore » 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

  5. Development of the Lens Antenna Deployment Demonstration (LADD) shuttle-attached flight experiment

    NASA Technical Reports Server (NTRS)

    Hill, H.; Johnston, D.; Frauenberger, H.

    1986-01-01

    The primary objective of the LADD Program is to develop a technology demonstration test article that can be used for both ground and flight tests to demonstrate the structural and mechanical feasibility and reliability of the single-axis roll-out space based radar (SBR) approach. As designed, the LADD will essentially be a generic strucutural experiment which incorporates all critical technology elements of the operational satellite and is applicable to a number of future antenna systems. However, to fully determine its design integrity for meeting the lens flatness and constant geometry requirements in a zero g environment under extreme thermal conditions, the LADD must be space flight tested. By accurately surveying the structure under varying conditions the membrane tolerance-holding capabilities of the structure will be demonstrated. The flight test will provide data to verify analytical tools used to predict thermal and structural behavior. Most important, the experiment will provide an initial indication of structural damping in a zero g vacuum environment. The recently completed Solar Array Flight Experiment (SAFE) showed orbital damping greater than that experienced during ground testing. From the experience and the information obtained from LADD it is hoped that designs can be confidently extrapolated to operational satellites with apertures in the 20 m by 60 m size range.

  6. 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.

  7. Development and demonstration of treatment technologies for the processing of US Department of Energy Mixed Waste

    SciTech Connect

    Bloom, G.A.; Berry, J.B.

    1994-01-01

    Mixed waste is defined as ``waste contaminated with chemically hazardous and radioactive species.`` The Mixed Waste Integrated Program (MWIP) was established in response to the need for a unified, DOE complexwide solution to issues of mixed waste treatment that meets regulatory requirements. MWIP is developing treatment technologies that reduce risk, minimize life-cycle cost, and improve process performance as compared to existing technologies. Treatment for waste streams for which no current technology exists, and suitable waste forms for disposal, will be provided to improve operations of the DOE Office of Waste Management. MWIP is composed of six technical areas within a mixed-waste treatment system: (1) systems analysis, (2) materials handling, (3) chemical/physical separation, (4) waste destruction and stabilization, (5) off-gas treatment, and (6) final waste form stabilization. The status of the technical initiatives and the current research, development, and demonstration in each of these areas are described in this paper

  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. Pit disassembly and conversion demonstration environmental assessment and research and development activities

    SciTech Connect

    1998-08-01

    A significant portion of the surplus plutonium is in the form of pits, a nuclear weapons component. Pits are composed of plutonium which is sealed in a metallic shell. These pits would need to be safely disassembled and permanently converted to an unclassified form that would be suitable for long-term disposition and international inspection. To determine the feasibility of an integrated pit disassembly and conversion system, a Pit Disassembly and Conversion Demonstration is proposed to take place at the Los Alamos National Laboratory (LANL). This demonstration would be done in existing buildings and facilities, and would involve the disassembly of up to 250 pits and conversion of the recovered plutonium to plutonium metal ingots and plutonium dioxide. This demonstration also includes the conversion of up to 80 kilograms of clean plutonium metal to plutonium dioxide because, as part of the disposition process, some surplus plutonium metal may be converted to plutonium dioxide in the same facility as the surplus pits. The equipment to be used for the proposed demonstration addressed in this EA would use some parts of the Advanced Recovery and Integrated Extraction System (ARIES) capability, other existing equipment/capacities, plus new equipment that was developed at other sites. In addition, small-scale R and D activities are currently underway as part of the overall surplus plutonium disposition program. These R and D activities are related to pit disassembly and conversion, MOX fuel fabrication, and immobilization (in glass and ceramic forms). They are described in Section 7.0. On May 16, 1997, the Office of Fissile Materials Disposition (MD) notified potentially affected states and tribes that this EA would be prepared in accordance with NEPA. This EA has been prepared to provide sufficient information for DOE to determine whether a Finding of No Significant Impact (FONSI) is warranted or whether an EIS must be prepared.

  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. 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.

  12. 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.

  13. 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

  14. 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.

  15. Molecular cloning and functional characterisation of VanX, a D-alanyl-D-alanine dipeptidase from Streptomyces coelicolor A3(2).

    PubMed

    Tan, Ai Lin; Loke, Paxton; Sim, Tiow-Suan

    2002-01-01

    The vanX gene which encodes a D-alanyl-D-alanine dipeptidase is critical for vancomycin resistance in enterococci. A putative vanX gene from Streptomyces coelicolor A3(2), which is not known for vancomycin production, was identified by homology-based analysis and cloned by polymerase chain reaction. The S. coelicolor vanX gene was heterologously expressed in Escherichia coli BL21(DE3) and enzymatic assays of soluble protein fractions of VanX revealed a 93-fold increase in dipeptidase activity as compared to the nonrecombinant control, thus confirming its functionality. Interestingly, S. coelicolor was also found to be of intermediate resistance to vancomycin although it does not produce vancomycin, thus suggesting the role of VanX in defence or immunity. As such, the prevalence of vanX genes in the environment may be more common than previously thought.

  16. Expression and displaying of β-glucosidase from Streptomyces coelicolor A3 in Escherichia coli.

    PubMed

    Gu, Ming-Zhu; Wang, Jing-Chao; Liu, Wei-Bing; Zhou, Ying; Ye, Bang-Ce

    2013-08-01

    Two genes encoding β-glucosidase from Streptomyces coelicolor A3(2) were cloned and expressed in Escherichia coli BL21 (DE3). Two recombinant enzymes (SC1059 and SC7558) were purified and characterized. The molecular mass of the purified SC1059 and SC7558 as determined by SDS-PAGE agrees with the calculated values (51.0 and 52.2 kDa, respectively). Optimal temperature and pH for the two enzymes were both at 35 °C and 6.0. SC7558 exhibited to be much more active than SC1059 under optimal conditions, and it was recombined with ice nucleation protein which could anchor on the surface of the cell. The optimal temperature and pH of the recombinant cells were 55 °C and 8.0, respectively. The resultant cells were to be used as material for immobilized β-glucosidase, which is convenient to catalyze substrates in various complicated conditions. PMID:23722947

  17. Conditional control of gene expression by synthetic riboswitches in Streptomyces coelicolor.

    PubMed

    Rudolph, Martin M; Vockenhuber, Michael-Paul; Suess, Beatrix

    2015-01-01

    Here we provide a step-by-step protocol for the application of synthetic theophylline-dependent riboswitches for conditional gene expression in Streptomyces coelicolor. Application of the method requires a sequence of only ~85 nt to be inserted between the transcriptional start site and the start codon of a gene of interest. No auxiliary factors are needed. All tested riboswitch variants worked well in concert with the promoters galP2, ermEp1, and SF14. Moreover, they allowed theophylline-dependent expression not only of the heterologous β-glucuronidase reporter gene but also of dagA, an endogenous agarase gene. The right combination of the tested promoters with the riboswitch variants allows for the adjustment of the desired dynamic range of regulation in a highly specific and dose-dependent manner and underlines the orthogonality of riboswitch regulation. We anticipate that any additional natural or synthetic promoter can be combined with the presented riboswitches. Moreover, this system should easily be transferable to other Streptomyces species, and most likely to any other genetically manipulable bacteria. PMID:25605391

  18. 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

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

    PubMed

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

    2015-10-01

    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

  20. 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.

  1. Industrial advanced turbine systems: Development and demonstration. Quarterly report, October 1--December 31, 1997

    SciTech Connect

    1998-06-01

    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 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 24.7% complete (22.8% last quarter). Work on the Mercury 50 development and ATS technology development portions of the program (WBS 10000 et seq) is 41.6% complete. Although a great amount of work occurred in the quarter, a significant amount of this work entailed the revision and rerelease of several Mercury 50 drawings. Estimates of percent compete 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 variation information is provided in Section 4.0 Program Management.

  2. 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.

  3. 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).

  4. Development and demonstration of a pilot-scale debris washing system

    SciTech Connect

    Taylor, M.L.; Barkley, N.P.

    1991-01-01

    Metallic, masonry, and other solid debris that may be contaminated with hazardous chemicals litter numerous hazardous waste sites in the United States. Polychlorinated biphenyls (PCB's), pesticides, lead or other metals are some of the contaminants of concern. In some cases cleanup standards have been established (e.g., 10 micrograms PCB's/100 sq cm for surfaces to which humans may be frequently exposed). Decontaminated debris could be either returned to the site as 'clean' fill, or, in the case of the metallic debris, sold to a metal smelter. The project involves the development and demonstration of a technology specifically for performing on-site decontamination of debris. Both bench-scale and pilot-scale versions of a debris washing system (DWS) have been designed, constructed and demonstrated. The DWS entails the application of an aqueous solution during a high-pressure spray cycle, followed by turbulent wash and rinse cycles. The aqueous cleaning solution is recovered and reconditioned for reuse concurrently with the debris-cleaning process, which minimizes the quantity of process water required to clean the debris.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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

  10. 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.

  11. 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

  12. Data-gathering method for use in modeling energy research, development, and demonstration programs

    SciTech Connect

    Meyer, M.A.; Booker, J.M.; Cullingford, H.S.; Peaslee, A.T. Jr.

    1981-01-01

    The development and testing of a data-gathering method for use in a computer program designed to model energy research, development, and demonstration programs for decisionmakers are described. The data-gathering method consists of face-to-face interviews with the scientists working on the projects that will be modeled by the computer program. The basic information gained from an interview includes time estimates for reaching certain project goals and the probability of achieving those goals within the times estimated. The interview method is based on decision analysis techniques. The Magnetic Fusion Energy program of the US Department of Energy was selected as the test case for this project. The data gathering method was used at five fusion projects to determine whether it could meet its design criteria. Extensive statistical analysis was performed to learn how much the expert's answers agreed, what factors were likely to enter into their estimates, and how their estimates corresponded to the officially scheduled dates and to the dates that the project goals were actually achieved. The interview method was considered to have met its design criteria and to be a valid tool for planning.

  13. 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.

  14. 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

  15. Heterologous expression of natural product biosynthetic gene clusters in Streptomyces coelicolor: from genome mining to manipulation of biosynthetic pathways.

    PubMed

    Gomez-Escribano, Juan Pablo; Bibb, Mervyn J

    2014-02-01

    Heterologous gene expression is one of the main strategies used to access the full biosynthetic potential of actinomycetes, as well as to study the metabolic pathways of natural product biosynthesis and to create unnatural pathways. Streptomyces coelicolor A3(2) is the most studied member of the actinomycetes, bacteria renowned for their prolific capacity to synthesize a wide range of biologically active specialized metabolites. We review here the use of strains of this species for the heterologous production of structurally diverse actinomycete natural products.

  16. 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

  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. 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.

  20. Development and demonstration flight of a fuel cell system for high-altitude balloons

    NASA Astrophysics Data System (ADS)

    Uno, Masatoshi; Shimada, Takanobu; Ariyama, Yusuke; Fukuzawa, Naoya; Noguchi, Daisuke; Ogawa, Keita; Naito, Hitoshi; Sone, Yoshitsugu; Saito, Yoshitaka

    Proton exchange membrane fuel cell offers higher energy density than the existing battery technologies for high-energy applications, and it is a promising power source for various industries including aerospace vehicles. We have been developing and testing a non-external humidified fuel cell system for high-altitude balloons, which require simple, light, and easy-to-operate power systems. This system consists of three major subsystems-a fuel cell stack, a reactant supply subsystem, and an electrical control subsystem. Ground performance testing in a vacuum chamber simulating the high-altitude vacuum condition was performed before the flight. Then, a demonstration flight of the fuel cell system was launched using a large balloon for verifying its performance under practical high-altitude conditions. Cell voltage variations synchronized with oxygen pressure spikes were observed that were probably caused by condensed product water plugging the flow passages of the back pressure regulator. Flight results indicated that the fuel cell system operated better when water was expelled as vapor, rather than in the liquid form. In addition, a back pressure regulator should be installed to avoid the accumulation of water droplets for realizing a stable performance.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. Comparative genomics of transport proteins in developmental bacteria: Myxococcus xanthus and Streptomyces coelicolor

    PubMed Central

    2013-01-01

    Background Two of the largest fully sequenced prokaryotic genomes are those of the actinobacterium, Streptomyces coelicolor (Sco), and the δ-proteobacterium, Myxococcus xanthus (Mxa), both differentiating, sporulating, antibiotic producing, soil microbes. Although the genomes of Sco and Mxa are the same size (~9 Mbp), Sco has 10% more genes that are on average 10% smaller than those in Mxa. Results Surprisingly, Sco has 93% more identifiable transport proteins than Mxa. This is because Sco has amplified several specific types of its transport protein genes, while Mxa has done so to a much lesser extent. Amplification is substrate- and family-specific. For example, Sco but not Mxa has amplified its voltage-gated ion channels but not its aquaporins and mechano-sensitive channels. Sco but not Mxa has also amplified drug efflux pumps of the DHA2 Family of the Major Facilitator Superfamily (MFS) (49 versus 6), amino acid transporters of the APC Family (17 versus 2), ABC-type sugar transport proteins (85 versus 6), and organic anion transporters of several families. Sco has not amplified most other types of transporters. Mxa has selectively amplified one family of macrolid exporters relative to Sco (16 versus 1), consistent with the observation that Mxa makes more macrolids than does Sco. Conclusions Except for electron transport carriers, there is a poor correlation between the types of transporters found in these two organisms, suggesting that their solutions to differentiative and metabolic needs evolved independently. A number of unexpected and surprising observations are presented, and predictions are made regarding the physiological functions of recognizable transporters as well as the existence of yet to be discovered transport systems in these two important model organisms and their relatives. The results provide insight into the evolutionary processes by which two dissimilar prokaryotes evolved complexity, particularly through selective chromosomal gene

  8. A system for the targeted amplification of bacterial gene clusters multiplies antibiotic yield in Streptomyces coelicolor.

    PubMed

    Murakami, Takeshi; Burian, Jan; Yanai, Koji; Bibb, Mervyn J; Thompson, Charles J

    2011-09-20

    Gene clusters found in bacterial species classified as Streptomyces encode the majority of known antibiotics as well as many pharmaceutically active compounds. A site-specific recombination system similar to those that mediate plasmid conjugation was engineered to catalyze tandem amplification of one of these gene clusters in a heterologous Streptomyces species. Three genetic elements were known to be required for DNA amplification in S. kanamyceticus: the oriT-like recombination sites RsA and RsB, and ZouA, a site-specific relaxase similar to TraA proteins that catalyze plasmid transfer. We inserted RsA and RsB sequences into the S. coelicolor genome flanking a cluster of 22 genes (act) responsible for biosynthesis of the polyketide antibiotic actinorhodin. Recombination between RsA and RsB generated zouA-dependent DNA amplification resulting in 4-12 tandem copies of the act gene cluster averaging nine repeats per genome. This resulted in a 20-fold increase in actinorhodin production compared with the parental strain. To determine whether the recombination event required taxon-specific genetic effectors or generalized bacterial recombination (recA), it was also analyzed in the heterologous host Escherichia coli. zouA was expressed under the control of an inducible promoter in wild-type and recA mutant strains. A plasmid was constructed with recombination sites RsA and RsB bordering a drug resistance marker. Induction of zouA expression generated hybrid RsB/RsA sites, evidence of site-specific recombination that occurred independently of recA. ZouA-mediated DNA amplification promises to be a valuable tool for increasing the activities of commercially important biosynthetic, degradative, and photosynthetic pathways in a wide variety of organisms.

  9. A system for the targeted amplification of bacterial gene clusters multiplies antibiotic yield in Streptomyces coelicolor

    PubMed Central

    Murakami, Takeshi; Burian, Jan; Yanai, Koji; Bibb, Mervyn J.; Thompson, Charles J.

    2011-01-01

    Gene clusters found in bacterial species classified as Streptomyces encode the majority of known antibiotics as well as many pharmaceutically active compounds. A site-specific recombination system similar to those that mediate plasmid conjugation was engineered to catalyze tandem amplification of one of these gene clusters in a heterologous Streptomyces species. Three genetic elements were known to be required for DNA amplification in S. kanamyceticus: the oriT-like recombination sites RsA and RsB, and ZouA, a site-specific relaxase similar to TraA proteins that catalyze plasmid transfer. We inserted RsA and RsB sequences into the S. coelicolor genome flanking a cluster of 22 genes (act) responsible for biosynthesis of the polyketide antibiotic actinorhodin. Recombination between RsA and RsB generated zouA-dependent DNA amplification resulting in 4–12 tandem copies of the act gene cluster averaging nine repeats per genome. This resulted in a 20-fold increase in actinorhodin production compared with the parental strain. To determine whether the recombination event required taxon-specific genetic effectors or generalized bacterial recombination (recA), it was also analyzed in the heterologous host Escherichia coli. zouA was expressed under the control of an inducible promoter in wild-type and recA mutant strains. A plasmid was constructed with recombination sites RsA and RsB bordering a drug resistance marker. Induction of zouA expression generated hybrid RsB/RsA sites, evidence of site-specific recombination that occurred independently of recA. ZouA-mediated DNA amplification promises to be a valuable tool for increasing the activities of commercially important biosynthetic, degradative, and photosynthetic pathways in a wide variety of organisms. PMID:21903924

  10. 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…

  11. 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…

  12. 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…

  13. Evaluation of Streptomyces coelicolor A3(2) as a heterologous expression host for the cyanobacterial protein kinase C activator lyngbyatoxin A.

    PubMed

    Jones, Adam C; Ottilie, Sabine; Eustáquio, Alessandra S; Edwards, Daniel J; Gerwick, Lena; Moore, Bradley S; Gerwick, William H

    2012-04-01

    Filamentous marine cyanobacteria are extremely rich sources of bioactive natural products and often employ highly unusual biosynthetic enzymes in their assembly. However, the current lack of techniques for stable DNA transfer into these filamentous organisms, combined with the absence of heterologous expression strategies for nonribosomal cyanobacterial gene clusters, prohibit the creation of mutant strains or the heterologous production of these cyanobacterial compounds in other bacteria. In this study, we evaluated the capability of a derivative of the model actinomycete Streptomyces coelicolor A3(2) to express enzymes involved in the biosynthesis of the protein kinase C activator lyngbyatoxin A from a Hawaiian strain of Moorea producta (previously classified as Lyngbya majuscula). Despite large differences in GC content between these two bacteria and the presence of rare TTA/UUA leucine codons in lyngbyatoxin ORFs we were able to achieve expression of the cytochrome P450 monooxygenase LtxB and reverse prenyltransferase LtxC in S. coelicolor M512 and confirmed the in vitro functionality of S. coelicolor overexpressed LtxC. Attempts to express the entire lyngbyatoxin A gene cluster in S. coelicolor M512 were not successful because of transcript termination observed for the ltxA gene, which encodes a large nonribosomal peptide synthetase. However, these attempts did show a detectable level of cyanobacterial promoter recognition in Streptomyces. Successful expression of lyngbyatoxin A proteins in Streptomyces provides a new platform for biochemical investigation of natural product enzymes from Moorea strains.

  14. Two X-38 Ship Demonstrators in Development at NASA Johnson Space Flight Center

    NASA Technical Reports Server (NTRS)

    1999-01-01

    This photo shows two X-38 Crew Return Vehicle technology demonstrators under development at NASA's Johnson Space Flight Center, Houston, Texas. The X-38 Crew Return Vehicle (CRV) research project is designed to develop the technology for a prototype emergency crew return vehicle, or lifeboat, for the International Space Station. The project is also intended to develop a crew return vehicle design that could be modified for other uses, such as a joint U.S. and international human spacecraft that could be launched on the French Ariane-5 Booster. The X-38 project is using available technology and off-the-shelf equipment to significantly decrease development costs. Original estimates to develop a capsule-type crew return vehicle were estimated at more than $2 billion. X-38 project officials have estimated that development costs for the X-38 concept will be approximately one quarter of the original estimate. Off-the-shelf technology is not necessarily 'old' technology. Many of the technologies being used in the X-38 project have never before been applied to a human-flight spacecraft. For example, the X-38 flight computer is commercial equipment currently used in aircraft and the flight software operating system is a commercial system already in use in many aerospace applications. The video equipment for the X-38 is existing equipment, some of which has already flown on the space shuttle for previous NASA experiments. The X-38's primary navigational equipment, the Inertial Navigation System/Global Positioning System, is a unit already in use on Navy fighters. The X-38 electromechanical actuators come from previous joint NASA, U.S. Air Force, and U.S. Navy research and development projects. Finally, an existing special coating developed by NASA will be used on the X-38 thermal tiles to make them more durable than those used on the space shuttles. The X-38 itself was an unpiloted lifting body designed at 80 percent of the size of a projected emergency crew return vehicle

  15. Development of Nb3Sn 11 T single aperture demonstrator dipole for LHC upgrades

    SciTech Connect

    Zlobin, A.V.; Apollinari, G.; Andreev, N.; Barzi, E.; Kashikhin, V.V.; Nobrega, f.; Novitski, I.; Auchmann, B.; Karppinen, M.; Rossi, L.; /CERN

    2011-03-01

    The LHC collimation upgrade foresees additional collimators installed in dispersion suppressor regions. To obtain the necessary space for the collimators, a solution based on the substitution of LHC main dipoles for stronger dipoles is being considered. CERN and FNAL have started a joint program to demonstrate the feasibility of Nb{sub 3}Sn technology for this purpose. The goal of the first phase is the design and construction of a 2-m long single-aperture demonstrator magnet with a nominal field of 11 T at 11.85 kA with 20% margin. This paper describes the magnetic and mechanical design of the demonstrator magnet and summarizes its design parameters.

  16. A Regulatory Gene SCO2140 is Involved in Antibiotic Production and Morphological Differentiation of Streptomyces coelicolor A3(2).

    PubMed

    Yu, Lingjun; Pan, Yuanyuan; Liu, Gang

    2016-08-01

    Streptomyces coelicolor is the soil-dwelling bacterium with a complex life cycle and a strong ability to produce plenty of secondary metabolites which are strictly regulated by a variety of regulators. Amino acid alignment shows that the deduced protein of SCO2140 belongs to the family of Leucine-responsive regulatory proteins (Lrps). Disruption of SCO2140 significantly decreased the yields of actinorhodin and calcium-dependent antibiotics, and the complemented strain restored the antibiotic productions to the wild-type level. In contrast, overexpression of SCO2140 increased the actinorhodin production. In agreement with it, the transcriptions of actII-ORF4 and cdaR remarkably reduced in the SCO2140 disruption mutant. The aerial mycelium formation of the SCO2140 disruption mutant was clearly delayed in R2YE medium due to the decrease of ramS expression while its complemented strain could restore the normal formation of aerial mycelia. These results indicated that SCO2140 was involved in antibiotic biosynthesis and morphological differentiation of Streptomyces coelicolor A3(2). PMID:27113590

  17. Phosphorylation of AfsR by multiple serine/threonine kinases in Streptomyces coelicolor A3(2).

    PubMed

    Sawai, Reiko; Suzuki, Ayano; Takano, Yuji; Lee, Ping-Chin; Horinouchi, Sueharu

    2004-06-01

    AfsK, a protein serine/threonine kinase, autophosphorylates on serine and threonine residues and phosphorylates serine and threonine residues of AfsR, a transcriptional activator for afsS involved in secondary metabolism in Streptomyces coelicolor A3(2). pkaG encoding a 592-amino-acid protein and SCD10.09 (named afsL) encoding a 580-amino-acid protein, both of which encode an AfsK-like protein, were transcribed throughout growth. PkaG with a histidine-tag and the kinase catalytic domain of PkaG, produced in Escherichia coli, autophosphorylated dominantly on threonine and slightly on serine residues. In addition, these proteins phosphorylated AfsR on threonine and serine residues. The catalytic domain of AfsL also autophosphorylated and phosphorylated AfsR, on threonine and serine residues in both cases. AfsR was thus found to be phosphorylated by multiple kinases. Disruption of the chromosomal pkaG gene resulted in slightly reduced production of the pigmented antibiotic actinorhodin. These findings, together with the presence of about 40 AfsK homologues and at least five AfsR homologues in S. coelicolor A3(2), suggest that the regulatory networks via eukaryotic-type protein phosphorylation are more diverse and versatile than we have expected.

  18. ScbR- and ScbR2-mediated signal transduction networks coordinate complex physiological responses in Streptomyces coelicolor

    PubMed Central

    Li, Xiao; Wang, Juan; Li, Shanshan; Ji, Junjie; Wang, Weishan; Yang, Keqian

    2015-01-01

    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. PMID:26442964

  19. Crystallization and preliminary X-ray crystallographic analyses of Nur, a nickel-responsive transcription regulator from Streptomyces coelicolor

    SciTech Connect

    An, Young Jun; Ahn, Bo-Eun; Roe, Jung-Hye; Cha, Sun-Shin

    2008-02-01

    A nickel uptake regulator (Nur) from Streptomyces coelicolor has been crystallized. A 2.4 Å native data set and a 3.0 Å Ni-MAD data set were collected using synchrotron radiation. Nickel ions serve in the correct folding and function of microbial enzymes implicated in metabolic processes. Although nickel ions are indispensable for the survival of cells, the intracellular level of nickel ions needs to be properly maintained as excessive levels of nickel ions are toxic. Nur, a nickel-uptake regulator belonging to the Fur family, is a nickel-responsive transcription factor that controls nickel homeostasis and antioxidative response in Streptomyces coelicolor. Nur was purified and crystallized at 295 K. A 2.4 Å native data set and a 3.0 Å Ni-MAD data set were collected using synchrotron radiation. The Nur crystals belong to space group P3{sub 1}, with unit-cell parameters a = b = 78.17, c = 50.39 Å. Assuming the presence of two molecules in the asymmetric unit, the solvent content is estimated to be about 54.7%.

  20. Crystallization and preliminary X-ray analysis of AbsC, a novel regulator of antibiotic production in Streptomyces coelicolor

    SciTech Connect

    Stevenson, Clare E. M.; Kock, Holger; Mootien, Saraspadee; Davies, Sîan C.; Bibb, Mervyn J.; Lawson, David M.

    2007-03-01

    A novel regulator of antibiotic production in S. coelicolor, AbsC, has been crystallized in space group P2{sub 1}2{sub 1}2{sub 1}. X-ray data to 2.25 Å resolution were collected on station PX 14.1 at Daresbury. Crystals of recombinant AbsC (subunit MW = 18 313 Da; 158 amino acids), a novel regulator of antibiotic production from Streptomyces coelicolor, were grown by vapour diffusion. The protein crystallizes in space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 43.53, b = 121.30, c = 143.75 Å. Native data to a resolution of 2.25 Å were recorded at station PX 14.1 (Daresbury) from a single crystal. Preliminary analysis of these data suggests that the asymmetric unit contains four copies of the AbsC monomer, giving an estimated solvent content of 47.0%. AbsC belongs to the MarR family of proteins that mediate ligand-responsive transcriptional control.

  1. 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

  2. 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)

  3. 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

  4. 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

  5. Molecular Characterization of Xylobiose- and Xylopentaose-Producing β-1,4-Endoxylanase SCO5931 from Streptomyces coelicolor A3(2).

    PubMed

    Enkhbaatar, Bolormaa; Lee, Chang-Ro; Hong, Young-Soo; Hong, Soon-Kwang

    2016-09-01

    Streptomyces coelicolor A3(2) sco5931 gene was predicted to encode a putative xylanase A, a 477 amino acid protein belonging to glycoside hydrolase family 10. The entire sco5931 coding region was cloned and overexpressed in Streptomyces lividans TK24. Mature SCO5931 protein comprising 436 amino acids (47 kDa) was purified by single-step gel filtration chromatography from culture broth after ammonium sulfate precipitation, with 25.8-fold purification and yield of 30.6 %. The purified protein displayed a pronounced activity toward beechwood xylan as a substrate, but no activity was detected toward carboxymethylcellulose, Avicel, galactan, barley β-glucan, and xyloglucan, demonstrating that SCO5931 is a substrate-specific xylanase. Optimal xylanase activity was observed at 60 °C and pH 6.0. The addition of metal ions or EDTA did not affect the xylanase activity, while 4 mM MnCl2 severely inhibited the enzyme, reducing its activity by 87 %. Kinetic parameters of SCO5931 toward beechwood xylan were determined (K m  = 0.24 mg/mL, V max  = 6.86 μM/min). Thin layer chromatography and mass spectrometry analyses of the beechwood xylan SCO5931 hydrolysis products were conducted. Product masses corresponded to sodium adducts of xylobiose (m/z 305.24) and xylopentaose (m/z 701.59), indicating that SCO5931 specifically cleaves the β-1,4 linkage of xylan to yield xylobiose and xylopentaose. PMID:27146990

  6. afsR is a pleiotropic but conditionally required regulatory gene for antibiotic production in Streptomyces coelicolor A3(2).

    PubMed

    Floriano, B; Bibb, M

    1996-07-01

    The N-terminal region of AfsR, a putative pleiotropic regulatory protein for antibiotic production in Streptomyces coelicolor A3(2), is homologous to RedD and Actil-ORF4, pathway-specific regulatory proteins required for the production of the antibiotics undecylprodigiosin (Red) and actinorhodin (Act), respectively. The recent identification of afsS, which lies immediately 3' of afsR and which stimulates antibiotic production when cloned at high copy number, questioned whether afsR was a pleiotropic regulatory gene. In this study we demonstrate that multiple copies of afsR can stimulate both Act and Red production and that, despite its homology, it cannot substitute for the pathway-specific regulatory genes. Moreover, an in-frame deletion that removed most of the afsR coding sequence resulted in loss of Act and Red production, and a marked reduction in the synthesis of the calcium-dependent antibiotic (CDA), but only under some (non-permissive) nutritional conditions. Although additional copies of afsR resulted in elevated levels of the actII-ORF4 and redD transcripts, transcription of the pathway-specific regulatory genes under non-permissive conditions was unaffected by deletion of afsR. While afsR may operate independently of the pathway-specific regulatory proteins to influence antibiotic production, the activity of ActII-ORF4 and of RedD under non-permissive conditions could depend on interaction with, or modification by, AfsR.

  7. 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.

  8. 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.

  9. 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

  10. 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

  11. 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.

  12. 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…

  13. 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.

  14. 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.

  15. 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.A.; Beard, C.A.; Bracht, R.R.; Buksa, J.J.; Chavez, W.; DeVolder, B.G.

    1994-10-01

    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 MW{sub t}.

  16. 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.

  17. 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.

  18. Graphite Isotope Ratio Method Development Report: Irradiation Test Demonstration of Uranium as a Low Fluence Indicator

    SciTech Connect

    Reid, B.D.; Gerlach, D.C.; Love, E.F.; McNeece, J.P.; Livingston, J.V.; Greenwood, L.R.; Petersen, S.L.; Morgan, W.C.

    1999-10-20

    This report describes an irradiation test designed to investigate the suitability of uranium as a graphite isotope ratio method (GIRM) low fluence indicator. GIRM is a demonstrated concept that gives a graphite-moderated reactor's lifetime production based on measuring changes in the isotopic ratio of elements known to exist in trace quantities within reactor-grade graphite. Appendix I of this report provides a tutorial on the GIRM concept.

  19. 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)

  20. 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…

  1. 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...

  2. Developing and Demonstrating Knowledge: Ability and Non-Ability Determinants of Learning and Performance

    ERIC Educational Resources Information Center

    Beier, Margaret E.; Campbell, Madeline; Crook, Amy E.

    2010-01-01

    Ability and non-ability traits were examined as predictors of learning, operationalized as the development of knowledge structure accuracy, and exam performance in a semester-long course. As predicted by investment theories of intellectual development, both cognitive ability and non-ability traits were important determinants of learning and exam…

  3. 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)

  4. 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.

  5. Development and demonstration of a reverse-osmosis energy-recovery device. Final report

    SciTech Connect

    Andeen, G.B.; Eid, J.C.

    1982-06-01

    An energy-recovery device for a seawater reverse-osmosis (RO) system was designed, fabricated, and tested. The device, comprised of a valving system, uses waste streams from an RO system to drive a pump which, in turn, sends additional feed flows to the RO elements. Test data demonstrated that efficiencies exceeding 95% can be expected, and, hence, energy consumption in a seawater RO desalination system can be decreased by 50%. Further, conversion may be decreased from 30% in order that membrane life is extended, and the size of the main pump and prime mover can be reduced by 50%. The device was subjected to almost 1000 hours (175,000 cycles) of testing, simulating its operation in a 4800 gallon/day seawater system, producing an outlet pressure of over 850 lbs/sq-in. for an inlet pressure of 750 lbs/sq-in. The device demonstrated an ability to self-start and to be controlled. An analysis of value and costs suggested that the device would be available to systems as small as 10,000 gallons/day. The results of this work confirm that significant improvements in the energy and economic performance of seawater desalination systems can be achieved.

  6. 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

  7. 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.

  8. Development of a learning module using a virtual environment to demonstrate EMG and telerobotic control principles.

    PubMed

    Patterson, P E

    2002-01-01

    A prototype system was developed for use as a teaching tool, allowing students to link EMG monitoring, data smoothing, robotic control, and neural network training within a rapid prototyping virtual environment (VE). The VE software allowed for the rapid development of scenarios and, when linked with EMG data input to a neural network, allowed the user to control an artificial world containing a virtual arm. Student teams then attempted to control the arm in the VE while performing the tasks by use of a neural network system they had specifically developed and trained using their own EMG signals. The results from their system were then translated into a form that enabled the control of a real robot. Students enjoyed the challenge and uniqueness of the module, and were enthusiastic about extending the concept to other areas of interest.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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).

  15. 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,…

  16. 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.

  17. RESEARCH, DEVELOPMENT, AND DEMONSTRATION IN ADULT TRAINING AND RETRAINING. FINAL REPORT.

    ERIC Educational Resources Information Center

    KAUFMAN, JACOB J.

    A COOPERATIVE COMMUNITY ACTION PROGRAM WAS INITIATED AND DEVELOPED IN THE MON-YOUGH REGION OF ALLEGHENY COUNTY, PENNSYLVANIA. TO CORRECT SOCIAL AND ECONOMIC NEEDS THROUGH COMPREHENSIVE ADULT TRAINING, RETRAINING, AND EMPLOYMENT PROJECTS. THE MON-YOUGH COMMUNITY ACTION COMMITTEE WAS FORMED THROUGH THE COOPERATIVE EFFORTS OF THE PENNSYLVANIA STATE…

  18. The Development and Demonstration of Multiple Regression Models for Operant Conditioning Questions.

    ERIC Educational Resources Information Center

    Fanning, Fred; Newman, Isadore

    Based on the assumption that inferential statistics can make the operant conditioner more sensitive to possible significant relationships, regressions models were developed to test the statistical significance between slopes and Y intercepts of the experimental and control group subjects. These results were then compared to the traditional operant…

  19. Implementation of the Stallings Classroom Management Staff Development Demonstration Project in Putnam County, West Virginia.

    ERIC Educational Resources Information Center

    Sullivan, Debra K.; And Others

    The primary objective of the Stallings Classroom Management Staff Development Model is to help teachers change so that they can manage their classrooms more effectively. In this model, classroom observations and specific recommendations for teaching behaviors are made for each teacher, and a series of teacher training sessions helps them to change…

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

    DOE PAGESBeta

    Barzi, E.; Andreev, N.; Li, P.; Lombardo, V.; Turrioni, D.; Zlobin, A. V.

    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.

  1. Partnerships for Improving Literacy in Urban Schools: Advanced Reading Development Demonstration Project

    ERIC Educational Resources Information Center

    Reading Teacher, 2008

    2008-01-01

    How many times have researchers and professional developers approached you or your school with "the answer" to improving literacy achievement? How did it work out? If changes occurred during the engagement with these outside partners, what remained after the project ended? Did such efforts lead to lasting change and improvement in student…

  2. 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...

  3. 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...

  4. Metabolic and evolutionary insights into the closely-related species Streptomyces coelicolor and Streptomyces lividans deduced from high-resolution comparative genomic hybridization

    PubMed Central

    2010-01-01

    Background Whilst being closely related to the model actinomycete Streptomyces coelicolor A3(2), S. lividans 66 differs from it in several significant and phenotypically observable ways, including antibiotic production. Previous comparative gene hybridization studies investigating such differences have used low-density (one probe per gene) PCR-based spotted arrays. Here we use new experimentally optimised 104,000 × 60-mer probe arrays to characterize in detail the genomic differences between wild-type S. lividans 66, a derivative industrial strain, TK24, and S. coelicolor M145. Results The high coverage and specificity (detection of three nucleotide differences) of the new microarrays used has highlighted the macroscopic genomic differences between two S. lividans strains and S. coelicolor. In a series of case studies we have validated the microarray and have identified subtle changes in genomic structure which occur in the Asp-activating adenylation domains of CDA non-ribosomal peptide synthetase genes which provides evidence of gene shuffling between these domains. We also identify single nucleotide sequence inter-species differences which exist in the actinorhodin biosynthetic gene cluster. As the glyoxylate bypass is non-functional in both S. lividans strains due to the absence of the gene encoding isocitrate lyase it is likely that the ethylmalonyl-CoA pathway functions as the alternative mechanism for the assimilation of C2 compounds. Conclusions This study provides evidence for widespread genetic recombination, rather than it being focussed at 'hotspots', suggesting that the previously proposed 'archipelago model' of genomic differences between S. coelicolor and S. lividans is unduly simplistic. The two S. lividans strains investigated differ considerably in genetic complement, with TK24 lacking 175 more genes than its wild-type parent when compared to S. coelicolor. Additionally, we confirm the presence of bldB in S. lividans and deduce that S. lividans 66

  5. 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.

  6. Crystallization and preliminary X-ray analysis of AbsC, a novel regulator of antibiotic production in Streptomyces coelicolor

    PubMed Central

    Stevenson, Clare E. M.; Kock, Holger; Mootien, Saraspadee; Davies, Sîan C.; Bibb, Mervyn J.; Lawson, David M.

    2007-01-01

    Crystals of recombinant AbsC (subunit MW = 18 313 Da; 158 amino acids), a novel regulator of antibiotic production from Streptomyces coelicolor, were grown by vapour diffusion. The protein crystallizes in space group P212121, with unit-cell parameters a = 43.53, b = 121.30, c = 143.75 Å. Native data to a resolution of 2.25 Å were recorded at station PX 14.1 (Daresbury) from a single crystal. Preliminary analysis of these data suggests that the asymmetric unit contains four copies of the AbsC monomer, giving an estimated solvent content of 47.0%. AbsC belongs to the MarR family of proteins that mediate ligand-responsive transcriptional control. PMID:17329821

  7. 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

  8. 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.

  9. 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.

  10. Nb3Sn cable development for the 11 T DS dipole demonstration model

    NASA Astrophysics Data System (ADS)

    Barzi, Emanuela; Lombardo, Vito; Nobrega, Alfred; Turrioni, Daniele; Yamada, Ryuji; Zlobin, Alexander; Karppinen, Mikko

    2012-06-01

    Fermilab (FNAL) and CERN have started the development of 11 T 11-m long Nb3Sn dipoles to replace a number of LHC NbTi dipoles and free space for cold collimators in the LHC DS areas. An important step in the design of these magnets is the development of the high aspecept ratio Nb3Sn cable to achieve the nominal field of 11 T at the nominal LHC operation current of 11.8 kA at 1.9 with 20% margin. Keystoned Cu wires and Nb3Sn RRP strand both mechanical stability and minimal damage to the restack design to be used in the magnet short models. Each cable was characterized electrically for transport properties degradation at high field and for low feild stability, and metallographically for internal damage.

  11. 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.

  12. 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

  13. 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)

  14. 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.

  15. Demonstration of anaerobic biogas digesters in developing countries. Part III. The Philippines

    SciTech Connect

    Simpson, M.H.; Morales, E.C.

    1980-03-01

    The main theme of this series of articles is that ours is now a world-wide society, short on meeting needs for energy yet long on waste from our industrial, agricultural and human consumption processes. This is a study report about developments in the Philippines where waste management has been recognized and considered as an important practical source of energy. This is revealed by several reports of the number of biogas plants in operation in this country. According to the July 31, 1977 survey made by the Philippines Bureau of Animal Industries, 200 biogas plants were then installed and in operation of which 46 were government-owned and 154 privately-owned. More have been installed since then. This report presents some of the operating observations and developments from the joint engineering analyses project of the Philippines Bureau of Animal Industry, Man and the Biosphere Inter-Agency Committee on Ecological Studies, Bureau of Fisheries and Aquatic Resources and the National Institute of Science and Technology. The project's main objective was to show that establishing a biogas plant involves not only the production of a methane gas mixture but the integration of its other products as part of a system (i.e., using effluent water from the biogas digester for production of algae chlorell sp. for livestock and poultry feed, production of fish and fertilizing-irrigating of pasture and vegetable plots.). Housing development sewer systems with added biogas generators are also discussed.

  16. 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."

  17. Global Gene Expression Profiling of Individual Human Oocytes and Embryos Demonstrates Heterogeneity in Early Development

    PubMed Central

    Zeef, Leo; Kimber, Susan J.; Brison, Daniel R.

    2013-01-01

    Early development in humans is characterised by low and variable embryonic viability, reflected in low fecundity and high rates of miscarriage, relative to other mammals. Data from assisted reproduction programmes provides additional evidence that this is largely mediated at the level of embryonic competence and is highly heterogeneous among embryos. Understanding the basis of this heterogeneity has important implications in a number of areas including: the regulation of early human development, disorders of pregnancy, assisted reproduction programmes, the long term health of children which may be programmed in early development, and the molecular basis of pluripotency in human stem cell populations. We have therefore investigated global gene expression profiles using polyAPCR amplification and microarray technology applied to individual human oocytes and 4-cell and blastocyst stage embryos. In order to explore the basis of any variability in detail, each developmental stage is replicated in triplicate. Our data show that although transcript profiles are highly stage-specific, within each stage they are relatively variable. We describe expression of a number of gene families and pathways including apoptosis, cell cycle and amino acid metabolism, which are variably expressed and may be reflective of embryonic developmental competence. Overall, our data suggest that heterogeneity in human embryo developmental competence is reflected in global transcript profiles, and that the vast majority of existing human embryo gene expression data based on pooled oocytes and embryos need to be reinterpreted. PMID:23717564

  18. 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.

  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. Gluconic acid-producing Pseudomonas sp. prevent γ-actinorhodin biosynthesis by Streptomyces coelicolor A3(2).

    PubMed

    Galet, Justine; Deveau, Aurélie; Hôtel, Laurence; Leblond, Pierre; Frey-Klett, Pascale; Aigle, Bertrand

    2014-09-01

    Streptomyces are ubiquitous soil bacteria well known for their ability to produce a wide range of secondary metabolites including antibiotics. In their natural environments, they co-exist and interact with complex microbial communities and their natural products are assumed to play a major role in mediating these interactions. Reciprocally, their secondary metabolism can be influenced by the surrounding microbial communities. Little is known about these complex interactions and the underlying molecular mechanisms. During pairwise co-culture experiments, a fluorescent Pseudomonas, Pseudomonas fluorescens BBc6R8, was shown to prevent the production of the diffusible blue pigment antibiotic γ-actinorhodin by Streptomyces coelicolor A3(2) M145 without altering the biosynthesis of the intracellular actinorhodin. A mutant of the BBc6R8 strain defective in the production of gluconic acid from glucose and consequently unable to acidify the culture medium did not show any effect on the γ-actinorhodin biosynthesis in contrast to the wild-type strain and the mutant complemented with the wild-type allele. In addition, when glucose was substituted by mannitol in the culture medium, P. fluorescens BBc6R8 was unable to acidify the medium and to prevent the biosynthesis of the antibiotic. All together, the results show that P. fluorescens BBc6R8 impairs the biosynthesis of the lactone form of actinorhodin in S. coelicolor by acidifying the medium through the production of gluconic acid. Other fluorescent Pseudomonas and the opportunistic pathogen Pseudomonas aeruginosa PAO1 also prevented the γ-actinorhodin production in a similar way. We propose some hypotheses on the ecological significance of such interaction.

  1. 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.

  2. 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.

  3. 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.

  4. 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

  5. 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)

  6. 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.

  7. 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.

  8. 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.

  9. 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. PMID:25223548

  10. Renewable energy for America's cities: Advanced Community Energy Systems Proposed Research, Development and Demonstration Program

    SciTech Connect

    Gleason, T.C.J.

    1993-01-01

    The first purpose of this paper is to describe ACES technologies and their potential impact on the environment, the US energy supply system and economy. The second purpose is to recommend an R,D D program to the US Department of Energy which has as its goal the rapid development of the most promising of the new technologies. ACES supply thermal energy to groups of buildings, communities and cities in the form of hot or chilled water for building space heating, domestic hot water or air conditioning. The energy is supplied via a network of insulated, underground pipes linking central sources of supply with buildings. ACES, by definition, employ very high energy efficiency conversion technologies such as cogeneration, heat pumps, and heat activated chillers. These systems also use renewable energy sources such as solar energy, winter cold, wind, and surface and subsurface warm and cold waters. ACES compose a new generation of community-scale building heating and air conditioning supply technologies. These new systems can effect a rapid and economical conversion of existing cities to energy supply by very efficient energy conversion systems and renewable energy systems. ACES technologies are the most promising near term means by which cities can make the transition from our present damaging dependence on fossil fuel supply systems to an economically and environmentally sustainable reliance on very high efficiency and renewable energy supply systems. When fully developed to serve an urban area, ACES will constitute a new utility system which can attain a level of energy efficiency, economy and reliance on renewable energy sources not possible with currently available energy supply systems.

  11. 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.

  12. 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.

  13. Roadmap for Research, Development, and Demonstration of Instrumentation, Controls, and Human-Machine Interface Technologies

    SciTech Connect

    Miller, Don W.; Arndt, Steven A.; Bond, Leonard J.; Dudenhoeffer, Donald D.; Hallbert, Bruce P.; Holcomb, David E.; Wood, Richard T.; Naser, Joseph A.; O'Hara, John M.; Quinn, Edward L.

    2008-06-01

    Instrumentation, controls, and human-machine interfaces are essential enabling technologies that strongly influence nuclear power plant performance and operational costs. The nuclear power industry is currently engaged in a transition from traditional analog-based instrumentation, controls, and human-machine interface (ICHMI) systems to implementations employing digital technologies. This transition has primarily occurred in an ad hoc fashion through individual system upgrades at existing plants and has been constrained by a number of concerns. Although international implementation of evolutionary nuclear power plants and the progression toward new plants in the United States have spurred design of more fully digital plant-wide ICHMI systems, the experience base in the nuclear power application domain is limited. Additionally, design and development programs by the U.S. Department of Energy (DOE) for advanced reactor concepts, such as the Generation IV Program and Next Generation Nuclear Plant (NGNP), introduce different plant conditions and unique plant configurations that increase the need for enhanced ICHMI capabilities to fully achieve programmatic goals related to economic competitiveness, safety and reliability, sustainability, and proliferation resistance and physical protection. As a result, there are challenges that need to be addressed to enable the nuclear power industry to effectively and efficiently complete the transition to safe and comprehensive use of digital technology.

  14. 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.

  15. 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.

  16. Project to develop and demonstrate methods to eliminate frozen-coal-handling problems. Final report

    SciTech Connect

    Not Available

    1982-11-01

    This paper represents an evaluation of methods of removing frozen coal from railroad hopper cars. The work was based on a careful study of previous methods and examination of innovative methods proposed. It was found that chemical freeze conditioning and side release additives helped for temperatures above 5/sup 0/F but were of marginal benefit for lower temperatures. Further, the effectiveness varied with the particular material and the amount and method of application, and was not well correlated with the price. An innovative method United Coal liked and continued development at their own expense involved an auger/air cannon system in which the auger penetrated the frozen crust to within 2 to 3 feet of the bottom hopper doors and then an air blast from a tank at 140 to 350 psi freed the coal. This was particularly effective when used in connection with the existing overhead shaker facility. Other methods were also evaluated. Some were too expensive in energy, damaged the cars, or were ineffective (or in some cases dangerous). (LTN)

  17. NsdB, a TPR-like-domain-containing protein negatively affecting production of antibiotics in Streptomyces coelicolor A3 (2).

    PubMed

    Zhang, Li; Li, Wen-Cheng; Zhao, Chun-Hua; Chater, Keith F; Tao, Mei-Feng

    2007-10-01

    Tetratricopeptide repeat (TPR) domains usually mediate protein-protein interactions. NsdA, one of the 70 proteins containing TPR-like domains in Streptomyces coelicolor A3 (2), was previously found to negatively control sporulation and antibiotic production. Here we show that elimination of SCO7252, which encodes another of these proteins, also caused overproduction of two antibiotics, actinorhodin and CDA, but did not affect morphological differentiation. Disruption of SCO1593, encoding another of the family, had no obvious phenotypic effects. In surface-grown cultures, expression of SCO7252, which was named nsdB, was initiated at about 30 h, like that of nsdA. Analysis in silico of the 70 predicted TPR-like-containing proteins of S. coelicolor showed that 32 of them contained only TPR-like domains, and 25 of the remainder contained additional DNA-binding domains, implying that they might control gene expression directly.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. 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.…

  3. Demonstrating Student Success: A Practical Guide to Outcomes-Based Assessment of Learning and Development in Student Affairs

    ERIC Educational Resources Information Center

    Bresciani, Marilee J.; Gardner, Megan Moore; Hickmott, Jessica

    2010-01-01

    This practical guide to outcomes-based assessment in student affairs is designed to help readers meet the growing demand for accountability and for demonstrating student learning. The authors offer a framework for implementing the assessment of student learning and development and pragmatic advice on the strategies most appropriate for the…

  4. 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

    ... adverse comments are received, the U.S. EPA will review those comments and publish another FR document... research, development and demonstration permits (69 FR 13242). This rule allows for variances from... outlined in 40 CFR Part 239.12. Ohio's MSWLF permit program was approved on June 13, 1994 (59 FR 112)....

  5. 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

  6. Forkhead‐associated proteins genetically linked to the serine/threonine kinase PknB regulate carbon flux towards antibiotic biosynthesis in Streptomyces coelicolor

    PubMed Central

    Jones, Greg; Del Sol, Ricardo; Dudley, Ed; Dyson, Paul

    2011-01-01

    Summary To date, the function of only two of the 34 predicted serine/threonine protein kinases (STPKs) of Streptomyces coelicolor has been described. Here we report functional analysis of pknB and two linked genes, fhaAB, encoding forkhead‐associated (FHA) domain proteins that are part of a highly conserved gene locus in actinobacteria. In contrast to the homologous gene of Mycobacterium tuberculosis, pknB in S. coelicolor is not essential and has no apparent role in defining cell shape. Phosphorylation of recombinant forms of both the full‐length protein and N‐terminal kinase domain suggest that PknB‐mediated signalling in S. coelicolor may be modulated by another factor(s). FhaAB are candidate interacting partners of PknB and loss of their function resulted in deregulation of central carbon metabolism, with carbon flux diverted to synthesis of the antibiotic actinorhodin. The substrate hyphae of the fhaAB mutant also exhibited an unusual cording morphology. The results indicate that inactivation of FHA ‘brake’ proteins can potentially amplify the function of STPKs and, in this case, provide a means to overproduce antibiotics. PMID:21342471

  7. 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

  8. Developer/Demonstrator Directory.

    ERIC Educational Resources Information Center

    Far West Lab. for Educational Research and Development, San Francisco, CA.

    The National Diffusion Network (NDN) was established to bring together representatives of federal, state, and local agencies to promote widespread adoption of new and innovative programs within and across state boundaries. The goal in publishing this directory is to help local educators solve problems through program improvements gleaned from…

  9. Air Force highly integrated photonics program: development and demonstration of an optically transparent fiber optic network for avionics applications

    NASA Astrophysics Data System (ADS)

    Whaley, Gregory J.; Karnopp, Roger J.

    2010-04-01

    The goal of the Air Force Highly Integrated Photonics (HIP) program is to develop and demonstrate single photonic chip components which support a single mode fiber network architecture for use on mobile military platforms. We propose an optically transparent, broadcast and select fiber optic network as the next generation interconnect on avionics platforms. In support of this network, we have developed three principal, single-chip photonic components: a tunable laser transmitter, a 32x32 port star coupler, and a 32 port multi-channel receiver which are all compatible with demanding avionics environmental and size requirements. The performance of the developed components will be presented as well as the results of a demonstration system which integrates the components into a functional network representative of the form factor used in advanced avionics computing and signal processing applications.

  10. 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.

  11. 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.

  12. The U.S. Department of Energy`s integrated gasification combined cycle research, development and demonstration program

    SciTech Connect

    Brdar, R.D.; Cicero, D.C.

    1996-07-01

    Historically, coal has played a major role as a fuel source for power generation both domestically and abroad. Despite increasingly stringent environmental constraints and affordable natural gas, coal will remain one of the primary fuels for producing electricity. This is due to its abundance throughout the world, low price, ease of transport an export, decreasing capital cost for coal-based systems, and the need to maintain fuel diversity. Recognizing the role coal will continue to play, the US Department of Energy (DOE) is working in partnership with industry to develop ways to use this abundant fuel resource in a manner that is more economical, more efficient and environmentally superior to conventional means to burn coal. The most promising of these technologies is integrated gasification combined cycle (IGCC) systems. Although IGCC systems offer many advantages, there are still several hurdles that must be overcome before the technology achieves widespread commercial acceptance. The major hurdles to commercialization include reducing capital and operating costs, reducing technical risk, demonstrating environmental and technical performance at commercial scale, and demonstrating system reliability and operability. Overcoming these hurdles, as well as continued progress in improving system efficiency, are the goals of the DOE IGCC research, development and demonstrate (RD and D) program. This paper provides an overview of this integrated RD and D program and describes fundamental areas of technology development, key research projects and their related demonstration scale activities.

  13. 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)

  14. 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)

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. 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)

  20. 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)

  1. 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.

  2. 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)

  3. EXCEDE technology development IV: demonstration of polychromatic contrast in vacuum at 1.2 λ/D

    NASA Astrophysics Data System (ADS)

    Sirbu, Dan; Thomas, Sandrine J.; Belikov, Ruslan; Lozi, Julien; Bendek, Eduardo; Pluzhnik, Eugene; Lynch, Dana H.; Hix, Troy; Zell, Peter; Schneider, Glenn; Guyon, Olivier

    2015-09-01

    This paper is the fourth in the series on the technology development for the EXCEDE (EXoplanetary Circumstellar Environments and Disk Explorer) mission concept, which in 2011 was selected by NASA's Explorer program for technology development (Category III). EXCEDE is a 0.7m space telescope concept operating with a Phase Induced Amplitude Apodization (PIAA) Coronagraph and a Deformable Mirror (DM) to create a "dark-hole" or a region of high-contrast starlight suppression at the focal plane to allow direct imaging of exoplanets. This will allow fundamental science in the form of direct detection and spatial resolution of low surface brightness circumstellar debris disks, and the direct imaging of giant planets with angular separations as close in as the habitable zone of the host star. Thus, EXCEDE can function as both a scientific and technological precursor for any mission capable of imaging exo-Earths. Previously, we have reported experimental results on the first milestone, the demonstration of EXCEDE contrast in monochromatic light in air and more recently in vacuum. In this paper, we report on the procedure and the experimental results obtained for our second milestone demonstration of the EXCEDE starlight suppression system carried in a vacuum chamber at the Lockheed Martin Advanced Technology Center. This includes high contrast performance demonstrations at 1.2 λD, which includes a lab demonstration of 1x10-5 median contrast between 1.2 and 2.0 λD simultaneously with 3x10-7 median contrast between 2 and 11 λD in 10% bandwidth polychromatic light centered at 650 nm for a single-sided dark zone. The results are stable and repeatable as demonstrated by three measurement runs with DM settings set from scratch and maintained on the best 90% out of the 1000 collected frames per run. We compare reduced experimental data with simulation results from modeling experimental limits.

  4. 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.

  5. 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.

  6. 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.

  7. 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 demonstrations. (CW)

  8. Development and Demonstration of a Method to Evaluate Bio-Sampling Strategies Using Building Simulation and Sample Planning Software

    PubMed Central

    Dols, W. Stuart; Persily, Andrew K.; Morrow, Jayne B.; Matzke, Brett D.; Sego, Landon H.; Nuffer, Lisa L.; Pulsipher, Brent A.

    2010-01-01

    In an effort to validate and demonstrate response and recovery sampling approaches and technologies, the U.S. Department of Homeland Security (DHS), along with several other agencies, have simulated a biothreat agent release within a facility at Idaho National Laboratory (INL) on two separate occasions in the fall of 2007 and the fall of 2008. Because these events constitute only two realizations of many possible scenarios, increased understanding of sampling strategies can be obtained by virtually examining a wide variety of release and dispersion scenarios using computer simulations. This research effort demonstrates the use of two software tools, CONTAM, developed by the National Institute of Standards and Technology (NIST), and Visual Sample Plan (VSP), developed by Pacific Northwest National Laboratory (PNNL). The CONTAM modeling software was used to virtually contaminate a model of the INL test building under various release and dissemination scenarios as well as a range of building design and operation parameters. The results of these CONTAM simulations were then used to investigate the relevance and performance of various sampling strategies using VSP. One of the fundamental outcomes of this project was the demonstration of how CONTAM and VSP can be used together to effectively develop sampling plans to support the various stages of response to an airborne chemical, biological, radiological, or nuclear event. Following such an event (or prior to an event), incident details and the conceptual site model could be used to create an ensemble of CONTAM simulations which model contaminant dispersion within a building. These predictions could then be used to identify priority area zones within the building and then sampling designs and strategies could be developed based on those zones. PMID:27134782

  9. 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)

  10. 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)

  11. 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)

  12. 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)

  13. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1988-01-01

    This article details two demonstrations involving color changes. Included are "Manganese Color Reactions" and "Flame Colors Demonstration." Include a list of materials needed, procedures, cautions, and results. (CW)

  14. 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)

  15. An Underground Storage Tank Integrated Demonstration report. Volume 1, Waste Characterization Data and Technology Development Needs Assessment

    SciTech Connect

    Quadrel, M.J.; Hunter, V.L.; Young, J.K.; Lini, D.C.; Goldberg, C.

    1993-04-01

    The Waste Characterization Data and Technology Development Needs Assessment provides direct support to the Underground Storage Tank Integrated Demonstration (UST-ID). Key users of the study`s products may also include individuals and programs within the US Department of Energy (DOE) Office of Technology Development (EM-50), the Office of Waste Operations (EM-30), and the Office of Environmental Restoration (EM-40). The goal of this work is to provide the UST-ID with a procedure for allocating funds across competing characterization technologies in a timely and defensible manner. It resulted in three primary products: 1. It organizes and summarizes information on underground storage tank characterization data needs. 2. It describes current technology development activity related to each need and flags areas where technology development may be beneficial. 3. It presents a decision process, with supporting software, for evaluating, prioritizing, and integrating possible technology development funding packages. The data presented in this document can be readily updated as the needs of the Waste Operations and Environmental Restoration programs mature and as new and promising technology development options emerge.

  16. Molecular characterization of Streptomyces coelicolor A(3) SCO6548 as a cellulose 1,4-β-cellobiosidase.

    PubMed

    Lim, Ju-Hyeon; Lee, Chang-Ro; Dhakshnamoorthy, Vijayalakshmi; Park, Jae Seon; Hong, Soon-Kwang

    2016-02-01

    Genomic sequencing analysis and previous studies have shown that there are eight genes in Streptomyces coelicolor A3(2) encoding putative cellulases. One of these genes, sco6548, was cloned into the Streptomyces/Escherichia coli shuttle vector pUWL201PW. The recombinant protein was successfully overexpressed in S. lividans TK24 under the control of the strong ermE promoter. Sco6548 was 1740 bp in length, and encoded a 579-amino acid-, 60.8-kDa protein with strong hydrolyzing activity toward Avicel and filter paper, yielding cellobiose as the final product. SCO6548 showed optimal activity at 50°C and pH 5. The Km values of SCO6548 toward Avicel and filter paper were 15.38 and 16.1 mg/mL, respectively. The Vmax values toward Avicel and filter paper were 0.432 and 0.084 μM/min, respectively. EDTA did not affect cellulase activity; however, several divalent cations, including Co(2+), Cu(2+), Ni(2+) and Mn(2+) (at 10 mM) had severe inhibitory effects on enzyme activity. Our analysis showed that SCO6548 is a cellulose 1,4-β-cellobiosidase that hydrolyzes cellulose into cellobiose. PMID:26712534

  17. pH shock induces overexpression of regulatory and biosynthetic genes for actinorhodin productionin Streptomyces coelicolor A3(2).

    PubMed

    Kim, Yoon Jung; Song, Jae Yang; Moon, Myung Hee; Smith, Colin P; Hong, Soon-Kwang; Chang, Yong Keun

    2007-10-01

    Actinorhodin production is markedly enhanced when an acidic pH shock is applied to a surface-grown culture of Streptomyces coelicolor A3(2). For an in-depth study of this phenomenon, transcriptional analyses using DNA microarrays and reverse transcription polymerase chain reaction and proteomic analysis were performed. Investigated were expression levels of the regulators and enzymes responsible for signal transduction and actinorhodin biosynthesis and enzymes involved in some major metabolic pathways. Regulators PkaG, AfsR, AfsS and/or another unidentified regulator and ActII-ORF4, in sequence, were observed to be activated by pH shock. In addition, a number of genes associated with actinorhodin production and secretion and the major central metabolic pathways investigated were observed to be upregulated with pH shock. Fatty acid degradation was particularly promoted by pH shock, while fatty acid biosynthesis was suppressed; it is envisaged that this enriches the precursor pool (acetyl-CoA) and building blocks for actinorhodin biosynthesis. Furthermore, glucose 6-phosphate dehydrogenases, initiating the pentose phosphate pathway, were highly activated by pH shock, enriching the reduced nicotinamide adenine dinucleotide phosphate (NADPH) pool for biosynthesis in general. It is deduced that these metabolic changes caused by pH shock have positively contributed to the stimulation of actinorhodin biosynthesis in a concerted manner.

  18. 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

  19. Structure of geranyl diphosphate C-methyltransferase from Streptomyces coelicolor and implications for the mechanism of isoprenoid modification†

    PubMed Central

    Köksal, Mustafa; Chou, Wayne K. W.; Cane, David E.; Christianson, David W.

    2012-01-01

    Geranyl diphosphate C-methyltransferase (GPPMT) from Streptomyces coelicolor A3(2) is the first methyltransferase discovered that modifies an acyclic isoprenoid diphosphate, geranyl diphosphate (GPP), to yield a non-canonical acyclic allylic diphosphate product, 2-methylgeranyl diphosphate, which serves as the substrate for a subsequent cyclization reaction catalyzed by a terpenoid cyclase, methylisoborneol synthase. Here, we report the crystal structures of GPPMT in complex with GPP or the substrate analogue geranyl-S-thiolodiphosphate (GSPP) along with S-adenosyl-l-homocysteine in the cofactor binding site, resulting from in situ demethylation of S-adenosyl-l-methionine, at 2.05 Å and 1.82 Å resolution, respectively. These structures suggest that both GPP and GSPP can undergo catalytic methylation in crystalline GPPMT, followed by dissociation of the isoprenoid product. S-adenosyl-l-homocysteine remains bound in the active site, however, and does not exchange with a fresh molecule of cofactor S-adenosyl-l-methionine. These structures provide important clues regarding the molecular mechanism of the reaction, especially with regard to the face of the 2,3 double bond of GPP that is methylated as well as the stabilization of the resulting carbocation intermediate through cation-π interactions. PMID:22455498

  20. 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.

  1. 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

  2. Comparative genomics of Streptomyces avermitilis, Streptomyces cattleya, Streptomyces maritimus and Kitasatospora aureofaciens using a Streptomyces coelicolor microarray system

    PubMed Central

    Hsiao, Nai-hua

    2007-01-01

    DNA/DNA microarray hybridization was used to compare the genome content of Streptomyces avermitilis, Streptomyces cattleya, Streptomyces maritimus and Kitasatospora aureofaciens with that of Streptomyces coelicolor A3(2). The array data showed an about 93% agreement with the genome sequence data available for S. avermitilis and also showed a number of trends in the genome structure for Streptomyces and closely related Kitasatospora. A core central region was well conserved, which might be predicted from previous research and this was linked to a low degree of gene conservation in the terminal regions of the linear chromosome across all four species. Between these regions there are two areas of intermediate gene conservation by microarray analysis where gene synteny is still detectable in S. avermitilis. Nonetheless, a range of conserved genes could be identified within the terminal regions. Variation in the genes involved in differentiation, transcription, DNA replication, etc. provides interesting insights into which genes in these categories are generally conserved and which are not. The results also provide target priorities for possible gene knockouts in a group of bacteria with a very large numbers of genes with unknown functions compared to most bacterial species. Electronic Supplementary Material The online version of this article (doi:10.1007/s10482-007-9175-1) contains supplementary material, which is available to authorized users. PMID:17588127

  3. 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)

  4. 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.

  5. 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.

  6. 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)

  7. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1988-01-01

    Describes two demonstrations for college level chemistry courses including: "Electrochemical Cells Using Sodium Silicate" and "A Simple, Vivid Demonstration of Selective Precipitation." Lists materials, preparation, procedures, and precautions. (CW)

  8. 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)

  9. 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)

  10. 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)

  11. 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)

  12. 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)

  13. 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)

  14. 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)

  15. 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

  16. Characterization of DNA Binding Sites of RokB, a ROK-Family Regulator from Streptomyces coelicolor Reveals the RokB Regulon

    PubMed Central

    Bekiesch, Paulina; Forchhammer, Karl; Apel, Alexander Kristian

    2016-01-01

    ROK-family proteins have been described to act either as sugar kinases or as transcriptional regulators. Few ROK-family regulators have been characterized so far and most of them are involved in carbon catabolite repression. RokB (Sco6115) has originally been identified in a DNA-affinity capturing approach as a possible regulator of the heterologously expressed novobiocin biosynthetic gene cluster in Streptomyces coelicolor M512. Interestingly, both, the rokB deletion mutants as well as its overexpressing mutants showed significantly reduced novobiocin production in the host strain S.coelicolor M512. We identified the DNA-binding site for RokB in the promoter region of the novobiocin biosynthetic genes novH-novW. It overlaps with the novH start codon which may explain the reduction of novobiocin production caused by overexpression of rokB. Bioinformatic screening coupled with surface plasmon resonance based interaction studies resulted in the discovery of five RokB binding sites within the genome of S. coelicolor. Using the genomic binding sites, a consensus motif for RokB was calculated, which differs slightly from previously determined binding motifs for ROK-family regulators. The annotations of the possible members of the so defined RokB regulon gave hints that RokB might be involved in amino acid metabolism and transport. This hypothesis was supported by feeding experiments with casamino acids and L-tyrosine, which could also explain the reduced novobiocin production in the deletion mutants. PMID:27145180

  17. Ligand-bound Structures and Site-directed Mutagenesis Identify the Acceptor and Secondary Binding Sites of Streptomyces coelicolor Maltosyltransferase GlgE*

    PubMed Central

    Syson, Karl; Stevenson, Clare E. M.; Miah, Farzana; Barclay, J. Elaine; Tang, Minhong; Gorelik, Andrii; Rashid, Abdul M.; Lawson, David M.; Bornemann, Stephen

    2016-01-01

    GlgE is a maltosyltransferase involved in α-glucan biosynthesis in bacteria that has been genetically validated as a target for tuberculosis therapies. Crystals of the Mycobacterium tuberculosis enzyme diffract at low resolution so most structural studies have been with the very similar Streptomyces coelicolor GlgE isoform 1. Although the donor binding site for α-maltose 1-phosphate had been previously structurally defined, the acceptor site had not. Using mutagenesis, kinetics, and protein crystallography of the S. coelicolor enzyme, we have now identified the +1 to +6 subsites of the acceptor/product, which overlap with the known cyclodextrin binding site. The sugar residues in the acceptor subsites +1 to +5 are oriented such that they disfavor the binding of malto-oligosaccharides that bear branches at their 6-positions, consistent with the known acceptor chain specificity of GlgE. A secondary binding site remote from the catalytic center was identified that is distinct from one reported for the M. tuberculosis enzyme. This new site is capable of binding a branched α-glucan and is most likely involved in guiding acceptors toward the donor site because its disruption kinetically compromises the ability of GlgE to extend polymeric substrates. However, disruption of this site, which is conserved in the Streptomyces venezuelae GlgE enzyme, did not affect the growth of S. venezuelae or the structure of the polymeric product. The acceptor subsites +1 to +4 in the S. coelicolor enzyme are well conserved in the M. tuberculosis enzyme so their identification could help inform the design of inhibitors with therapeutic potential. PMID:27531751

  18. Glycosylation of a Streptomyces coelicolor A3(2) cell envelope protein is required for infection by bacteriophage phi C31.

    PubMed

    Cowlishaw, D A; Smith, M C

    2001-08-01

    Mutants of Streptomyces coelicolor A3(2) J1929 (Delta pglY) were isolated that were resistant to the Streptomyces temperate phage phi C31. These strains could be transfected with phi C31 DNA, but could not act as infective centres after exposure to phage. Thus, it was concluded that infection was blocked at the adsorption/DNA injection step. The mutants fell into three classes. Class I mutants were complemented by a gene, SCE87.05, isolated from the cosmid library of S. coelicolor A3(2). The product of SCE87.05 had good overall homology to a Mycobacterium tuberculosis hypothetical protein and regions with similarity to dolichol phosphate-D-mannose:protein O-D-mannosyltransferases. Concanavalin A (ConA) inhibited phi C31 infection of S. coelicolor J1929, and this could be partially reversed by the addition of the sugar, alpha-D-methyl-pyranoside. Moreover, glycosylated proteins from J1929, but not from the class I mutant DT1017, were detected using ConA as a probe in Western blots. Class I and II mutants were sensitive to phi C31hc, a previously isolated phage exhibiting an extended host range phenotype, conferred by h. A phage with the same phenotype, phi DT4002, was isolated independently, and a missense mutation was found in a putative tail gene. It is proposed that the phi C31 receptor is a cell wall glycoprotein, and that the phi C31h mutation compensates for the lack of glycosylation of the receptor.

  19. 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

  20. Iron-regulatory proteins DmdR1 and DmdR2 of Streptomyces coelicolor form two different DNA-protein complexes with iron boxes.

    PubMed Central

    Flores, Francisco J; Martín, Juan F

    2004-01-01

    In high G+C Gram-positive bacteria, the control of expression of genes involved in iron metabolism is exerted by a DmdR [divalent (bivalent) metal-dependent regulatory protein] in the presence of Fe2+ or other bivalent ions. The dmdR1 and dmdR2 genes of Streptomyces coelicolor were overexpressed in Escherichia coli and the DmdR1 and DmdR2 proteins were purified to homogeneity. Electrophoretic mobility-shift assays showed that both DmdR1 and DmdR2 bind to the 19-nt tox and desA iron boxes forming two different complexes in each case. Increasing the concentrations of DmdR1 or DmdR2 protein shifted these complexes from their low-molecular-mass form to the high-molecular-mass complexes. Formation of the DNA-protein complexes was prevented by the bivalent metal chelating agent 2,2'-dipyridyl and by antibodies specific against the DmdR proteins. Cross-linking with glutaraldehyde of pure DmdR1 or DmdR2 proteins showed that DmdR1 forms dimers, whereas DmdR2 is capable of forming dimers and probably tetramers. Ten different iron boxes were found in a search for iron boxes in the genome of S. coelicolor. Most of them correspond to putative genes involved in siderophore biosynthesis. Since the nucleotide sequence of these ten boxes is identical (or slightly different) with the synthetic DNA fragment containing the desA box used in the present study, it is proposed that DmdR1 and DmdR2 bind to the iron boxes upstream of at least ten different genes in S. coelicolor. PMID:14960152

  1. 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)

  2. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1986-01-01

    Describes a demonstration involving the controlled combustion of a mixture of metals with black and smokeless powder in a small Erlenmeyer flask. Also describes demonstrations using a device that precludes breathing of hazardous vapors during class demonstrations; the device is easy to transport and use in rooms without sinks. (JN)

  3. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1986-01-01

    Describes two demonstrations used in laboratory chemistry courses. Discusses a "pH-activated" display used to chemically and visually supplement lecture demonstrations. Outlines another demonstration designed to show that copper(II) chloride is made of two ions, blue and yellow, which are combined to produce green. (TW)

  4. 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)

  5. 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)

  6. 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)

  7. 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

  8. Development, Demonstration, and Device Physics of Fet-Accessed One-Transistor Gallium Arsenide Dynamic Memory Technologies

    NASA Astrophysics Data System (ADS)

    Neudeck, Philip Gerold

    The introduction of digital GaAs into modern high -speed computing systems has led to an increasing demand for high-density memory in these GaAs technologies. To date, most of the memory development efforts in GaAs have been directed toward four- and six-transistor static RAM's, which consume substantial chip area and dissipate much static power resulting in limited single-chip GaAs storage capacities. As it has successfully done in silicon, a one-transistor dynamic RAM approach could alleviate these problems making higher density GaAs memories possible. This dissertation discusses theoretical and experimental work that presents the possibility for a high-speed, low-power, one-transistor dynamic RAM technology in GaAs. The two elements of the DRAM cell, namely the charge storage capacitor and the access field-effect transistor have been studied in detail. Isolated diode junction charge storage capacitors have demonstrated 30 minutes of storage time at room temperature with charge densities comparable to those obtained in planar silicon DRAM capacitors. GaAs JFET and MESFET technologies have been studied, and with careful device design and choice of proper operating voltages experimental results show that both can function as acceptable access transistors. One-transistor MESFET- and JFET-accessed DRAM cells have been fabricated and operated at room temperature and above with a standby power dissipation that is only a small fraction of the power dissipated by the best commercial GaAs static RAM cells. A 2 x 2 bit demonstration array was built and successfully operated at room temperature to demonstrate the addressable read/write capability of this new technology.

  9. Development of a risk management tool for prioritizing chemical hazard-food pairs and demonstration for selected mycotoxins.

    PubMed

    Stornetta, Alessia; Engeli, Barbara E; Zarn, Jürg A; Gremaud, Gérard; Sturla, Shana J

    2015-07-01

    We developed a simple tool for ranking chemical hazard-food pairs to assist policy makers and risk managers selecting the hazard-food pairs that deserve more attention and need to be monitored during food safety inspections. The tool is based on the derivation of a "Priority Index" (PI) that results from the ratio of the potency of the hazard and the consumer exposure. The potency corresponds to a toxicity reference value of the hazard, whereas the exposure results from the combination of the concentration of the hazard in the food, and the food consumption. Tool's assumptions and limitations are demonstrated and discussed by ranking a dataset of 13 mycotoxins in 26 food items routinely analyzed in Switzerland. The presented ranking of mycotoxin-food pairs has to be considered as relative due to scarce exposure data availability, and uncertainties in toxicity reference values. However, this representative example allows demonstrating the simplicity and the ability of the PI tool to prioritize chemical hazard-food pairs.

  10. 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.

  11. 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.

  12. Development and demonstration of Microcomputer Intelligence for Technical Training (MITT). Final technical paper, July 1987-January 1988

    SciTech Connect

    Johnson, W.B.; Hunt, R.M.; Duncan, P.C.; Norton, J.E.

    1988-08-01

    The Microcomputer Intelligence for Technical Training research described here is founded upon a proven intelligent diagnostic training simulation to construct enhanced models of the student, instructor, and expert in a training environment. An off-the-shelf microcomputer was used to deliver an operational intelligent tutoring system (ITS) with 180 days of project start. The demonstration ITS was developed in cooperation with Air Force and NASA personnel at the Lyndon B. Johnson Space Center in Houston, Texas. The space-shuttle fuel cell was the technical domain for this first application. Initial trial use by astronauts, flight controllers, and technical-training personnel indicates that the approach is technically feasible and has instructional value for space training applications.

  13. 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.

  14. Tachykinin-Immunoreactive Neurons in Developing Feline Neostriatum: Somatodendritic Morphogenesis Demonstrated by Combined Immunohistochemistry/Golgi Impregnation-Gold Toning

    PubMed Central

    Fisher, Robin; Boylan, Marea

    2011-01-01

    This investigation was designed to survey and characterize the development of a key link between chemically mediated neurotransmission and neuronal cytoarchitecture in mammalian basal ganglia. Peroxidase immunohistochemical and Golgi impregnation/gold toning methods were combined to doubly label the tachykinin neuromodulator signature and somatodendritic structure of neostriatal neurons in late fetal, postnatal and adult cats. The results supported 3 conclusions of considerable significance. (1) Colocalization of immunohistochemical and Golgi impregnation/gold toning labels is a feasible, rational and productive means to identify and determine the somatodendritic morphogenesis of tachykinin neurons. (2) The application of this method to developing feline neostriatum demonstrates directly that the principal tachykinin cells are medium-sized spiny neurons, which undergo progressive growth and elaboration of cell bodies, dendritic arbors and dendritic spines during the late fetal and postnatal periods. (3) There is a strong but incomplete concordance between tachykinin and medium-sized spiny neuronal phenotypes, because a minor variant of medium-sized spiny neurons and rare subgroups of medium- and large-sized sparse spiny neurons also show the tachykinin neuromodulator signature. Taken together, these results suggest that neostriatal neurons show an early commitment to heterogeneous tachykinin phenotypes, although the full and final expression of their somatodendritic characteristics coincides with synaptogenesis. PMID:21346326

  15. 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.

  16. 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

  17. Oxygen-Dependent Control of Respiratory Nitrate Reduction in Mycelium of Streptomyces coelicolor A3(2)

    PubMed Central

    Fischer, Marco; Falke, Dörte; Pawlik, Tony

    2014-01-01

    Several members of the obligately aerobic genus Streptomyces are able to reduce nitrate, catalyzed by Nar-type respiratory nitrate reductases. A unique feature of Streptomyces coelicolor A3(2) compared with other streptomycetes is that it synthesizes three nonredundant Nar enzymes. In this study, we show that Nar2 is the main Nar enzyme active in mycelium and could characterize the conditions governing its synthesis. Nar2 was present at low levels in aerobically cultivated mycelium, but synthesis was induced when cultures were grown under oxygen limitation. Growth in the presence of high oxygen concentrations prevented the induction of Nar2 synthesis. Equally, an abrupt shift from aerobiosis to anaerobiosis did not result in the immediate induction of Nar2 synthesis. This suggests that the synthesis of Nar2 is induced during a hypoxic downshift, probably to allow maintenance of a proton gradient during the transition to anaerobiosis. Although no Nar2 could be detected in freshly harvested mature spores, synthesis of the enzyme could be induced after long-term (several days) incubation of these resting spores under anaerobic conditions. Induction of Nar2 synthesis in spores was linked to transcriptional control. Nar2 activity in whole mycelium was strictly dependent on the presence of a putative nitrate transporter, NarK2. The oxygen-dependent inhibition of nitrate reduction by Nar2 was mediated by NarK2-dependent nitrate:nitrite antiport. This antiport mechanism likely prevents the accumulation of toxic nitrite in the cytoplasm. A deletion of the narK2 gene had no effect on Nar1-dependent nitrate reduction in resting spores. Together, our results indicate redox-dependent transcriptional and posttranslational control of nitrate reduction by Nar2. PMID:25225271

  18. Convergent Transcription in the Butyrolactone Regulon in Streptomyces coelicolor Confers a Bistable Genetic Switch for Antibiotic Biosynthesis

    PubMed Central

    Chatterjee, Anushree; Drews, Laurie; Mehra, Sarika; Takano, Eriko; Kaznessis, Yiannis N.; Hu, Wei-Shou

    2011-01-01

    cis-encoded antisense RNAs (cis asRNA) have been reported to participate in gene expression regulation in both eukaryotic and prokaryotic organisms. Its presence in Streptomyces coelicolor has also been reported recently; however, its role has yet to be fully investigated. Using mathematical modeling we explore the role of cis asRNA produced as a result of convergent transcription in scbA-scbR genetic switch. scbA and scbR gene pair, encoding repressor–amplifier proteins respectively, mediates the synthesis of a signaling molecule, the γ-butyrolactone SCB1 and controls the onset of antibiotic production. Our model considers that transcriptional interference caused by convergent transcription of two opposing RNA polymerases results in fatal collision and transcriptional termination, which suppresses transcription efficiency. Additionally, convergent transcription causes sense and antisense interactions between complementary sequences from opposing strands, rendering the full length transcript inaccessible for translation. We evaluated the role of transcriptional interference and the antisense effect conferred by convergent transcription on the behavior of scbA-scbR system. Stability analysis showed that while transcriptional interference affects the system, it is asRNA that confers scbA-scbR system the characteristics of a bistable switch in response to the signaling molecule SCB1. With its critical role of regulating the onset of antibiotic synthesis the bistable behavior offers this two gene system the needed robustness to be a genetic switch. The convergent two gene system with potential of transcriptional interference is a frequent feature in various genomes. The possibility of asRNA regulation in other such gene-pairs is yet to be examined. PMID:21765930

  19. Structure of Epi-Isozizaene Synthase from Streptomyces coelicolor A3(2), a Platform for New Terpenoid Cyclization Templates

    SciTech Connect

    Aaron, Julie A.; Lin, Xin; Cane, David E.; Christianson, David W.

    2010-06-21

    The X-ray crystal structure of recombinant epi-isozizaene synthase (EIZS), a sesquiterpene cyclase from Streptomyces coelicolor A3(2), has been determined at 1.60 {angstrom} resolution. Specifically, the structure of wild-type EIZS is that of its closed conformation in complex with three Mg{sup 2+} ions, inorganic pyrophosphate (PP{sub i}), and the benzyltriethylammonium cation (BTAC). Additionally, the structure of D99N EIZS has been determined in an open, ligand-free conformation at 1.90 {angstrom} resolution. Comparison of these two structures provides the first view of conformational changes required for substrate binding and catalysis in a bacterial terpenoid cyclase. Moreover, the binding interactions of BTAC may mimic those of a carbocation intermediate in catalysis. Accordingly, the aromatic rings of F95, F96, and F198 appear to be well-oriented to stabilize carbocation intermediates in the cyclization cascade through cation-{pi} interactions. Mutagenesis of aromatic residues in the enzyme active site results in the production of alternative sesquiterpene product arrays due to altered modes of stabilization of carbocation intermediates as well as altered templates for the cyclization of farnesyl diphosphate. Accordingly, the 1.64 {angstrom} resolution crystal structure of F198A EIZS in a complex with three Mg{sup 2+} ions, PP{sub i}, and BTAC reveals an alternative binding orientation of BTAC; alternative binding orientations of a carbocation intermediate could lead to the formation of alternative products. Finally, the crystal structure of wild-type EIZS in a complex with four Hg{sup 2+} ions has been determined at 1.90 {angstrom} resolution, showing that metal binding triggers a significant conformational change of helix G to cap the active site.

  20. 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,…

  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.

    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)

  3. 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)

  4. 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)

  5. 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.…

  6. 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)

  7. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1990-01-01

    Described are demonstrations designed to reveal the important "nonsolvent" properties of water through its interaction with a toy called "Magic Sand" and other synthetic silica derivatives, especially those bonded with organic moities. The procedures for seven demonstrations along with a discussion of the effects are presented. (CW)

  8. 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…

  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.

    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)

  11. 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)

  12. 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…

  13. Complete Demonstration.

    ERIC Educational Resources Information Center

    Yelon, Stephen; Maddocks, Peg

    1986-01-01

    Describes four-step approach to educational demonstration: tell learners they will have to perform; what they should notice; describe each step before doing it; and require memorization of steps. Examples illustrate use of this process to demonstrate a general mental strategy, and industrial design, supervisory, fine motor, and specific…

  14. 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)

  15. 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)

  16. 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.…

  17. 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)

  18. Characterization of a New GlnR Binding Box in the Promoter of amtB in Streptomyces coelicolor Inferred a PhoP/GlnR Competitive Binding Mechanism for Transcriptional Regulation of amtB

    PubMed Central

    Wang, Ying; Cen, Xu-Feng

    2012-01-01

    The transcription of amtB in Streptomyces coelicolor has been proposed to be counter-regulated by GlnR (a global regulator for nitrogen metabolism) and PhoP (a global regulator for phosphate metabolism). However, the GlnR-protected region, which was deduced to be two 22-bp GlnR binding boxes (gTnAc-n6-GaAAc-n6-GtnAC-n6-GAAAc-n6, abbreviated as a1-b1 and a2-b2), was separated from the PhoP-protected region in the promoter of amtB, leaving the mechanism for this regulation undefined. In this study, another 22-bp GlnR binding box, which consisted of a3-site-n6-b3-site (a3-b3) overlapping with the PhoP-binding sequences, was identified in the promoter region of amtB by a DNase I footprinting assay. An electrophoretic mobility shift assay (EMSA) using purified recombinant GlnR and the synthetic amtB promoter fragments with the three GlnR binding boxes individually mutated demonstrated that every box was involved in GlnR binding in vitro. Further in vivo assays using the egfp reporter gene fused to various kinds of mutated promoter regions of amtB demonstrated that all of the three GlnR binding boxes were required for GlnR-mediated activation of amtB transcription under the nitrogen-limited condition. The results of EMSA using the amtB promoter with mixtures of recombinant His-tagged GlnR and Trx-His-S-tagged PhoP inferred that PhoP might compete against GlnR from binding at the a3-b3 site, attributable to the PhoP/GlnR counter-regulatory function subjected to further experimental proof. PMID:22821977

  19. 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

  20. Direct cloning and heterologous expression of the salinomycin biosynthetic gene cluster from Streptomyces albus DSM41398 in Streptomyces coelicolor A3(2).

    PubMed

    Yin, Jia; Hoffmann, Michael; Bian, Xiaoying; Tu, Qiang; Yan, Fu; Xia, Liqiu; Ding, Xuezhi; Stewart, A Francis; Müller, Rolf; Fu, Jun; Zhang, Youming

    2015-01-01

    Linear plus linear homologous recombination-mediated recombineering (LLHR) is ideal for obtaining natural product biosynthetic gene clusters from pre-digested bacterial genomic DNA in one or two steps of recombineering. The natural product salinomycin has a potent and selective activity against cancer stem cells and is therefore a potential anti-cancer drug. Herein, we separately isolated three fragments of the salinomycin gene cluster (salO-orf18) from Streptomyces albus (S. albus) DSM41398 using LLHR and assembled them into intact gene cluster (106 kb) by Red/ET and expressed it in the heterologous host Streptomyces coelicolor (S. coelicolor) A3(2). We are the first to report a large genomic region from a Gram-positive strain has been cloned using LLHR. The successful reconstitution and heterologous expression of the salinomycin gene cluster offer an attractive system for studying the function of the individual genes and identifying novel and potential analogues of complex natural products in the recipient strain. PMID:26459865

  1. SCO4008, a putative TetR transcriptional repressor from Streptomyces coelicolor A3(2), regulates transcription of sco4007 by multidrug recognition.

    PubMed

    Hayashi, Takeshi; Tanaka, Yoshikazu; Sakai, Naoki; Okada, Ui; Yao, Min; Watanabe, Nobuhisa; Tamura, Tomohiro; Tanaka, Isao

    2013-09-23

    SCO4008 from Streptomyces coelicolor A3(2) is a member of the TetR family. However, its precise function is not yet clear. In this study, the crystal structure of SCO4008 was determined at a resolution of 2.3Å, and its DNA-binding properties were analyzed. Crystal structure analysis showed that SCO4008 forms an Ω-shaped homodimer in which the monomer is composed of an N-terminal DNA-binding domain containing a helix-turn-helix and a C-terminal dimerization and regulatory domain possessing a ligand-binding cavity. The genomic systematic evolution of ligands by exponential enrichment and electrophoretic mobility shift assay revealed that four SCO4008 dimers bind to the two operator regions located between sco4008 and sco4007, a secondary transporter belonging to the major facilitator superfamily. Ligand screening analysis showed that SCO4008 recognizes a wide range of structurally dissimilar cationic and hydrophobic compounds. These results suggested that SCO4008 is a transcriptional repressor of sco4007 responsible for the multidrug resistance system in S. coelicolor A3(2).

  2. Epigenetic Activation of Antibacterial Property of an Endophytic Streptomyces coelicolor Strain AZRA 37 and Identification of the Induced Protein Using MALDI TOF MS/MS.

    PubMed

    Kumar, Jitendra; Sharma, Vijay K; Singh, Dheeraj K; Mishra, Ashish; Gond, Surendra K; Verma, Satish K; Kumar, Anuj; Kharwar, Ravindra Nath

    2016-01-01

    The endophytic Streptomyces coelicolor strain AZRA 37 was isolated from the surface sterilized root of Azadirachta indica A. Juss., commonly known as neem plant in India. Since only a few reports are available regarding epigenetic modulations of microbial entities, S. coelicolor was treated with different concentrations of 5-azacytidine for this purpose and evaluated for its antibacterial potential against five human pathogenic bacteria (Aeromonas hydrophila IMS/GN11, Enterococcus faecalis IMS/GN7, Salmonella typhi MTCC 3216, Shigella flexneri ATCC 12022 and Staphylococcus aureus ATCC 25923). The crude extract obtained from cultures treated with 25 μM concentration of 5-azacytidine, was found effective against all five pathogenic bacteria tested while the untreated control was only active against 3 pathogenic bacteria. HPLC analysis of crude compounds from treated cultures showed a greater number of compounds than that of the control. Extraction of whole cell protein and its SDS PAGE analysis showed an additional major protein band in 25 μM 5-azacytidine treated culture and MALDI TOF MS/MS analysis revealed that this protein belongs to the porin family. PMID:26844762

  3. Direct cloning and heterologous expression of the salinomycin biosynthetic gene cluster from Streptomyces albus DSM41398 in Streptomyces coelicolor A3(2)

    PubMed Central

    Yin, Jia; Hoffmann, Michael; Bian, Xiaoying; Tu, Qiang; Yan, Fu; Xia, Liqiu; Ding, Xuezhi; Francis Stewart, A.; Müller, Rolf; Fu, Jun; Zhang, Youming

    2015-01-01

    Linear plus linear homologous recombination-mediated recombineering (LLHR) is ideal for obtaining natural product biosynthetic gene clusters from pre-digested bacterial genomic DNA in one or two steps of recombineering. The natural product salinomycin has a potent and selective activity against cancer stem cells and is therefore a potential anti-cancer drug. Herein, we separately isolated three fragments of the salinomycin gene cluster (salO-orf18) from Streptomyces albus (S. albus) DSM41398 using LLHR and assembled them into intact gene cluster (106 kb) by Red/ET and expressed it in the heterologous host Streptomyces coelicolor (S. coelicolor) A3(2). We are the first to report a large genomic region from a Gram-positive strain has been cloned using LLHR. The successful reconstitution and heterologous expression of the salinomycin gene cluster offer an attractive system for studying the function of the individual genes and identifying novel and potential analogues of complex natural products in the recipient strain. PMID:26459865

  4. Metabolic Switches and Adaptations Deduced from the Proteomes of Streptomyces coelicolor Wild Type and phoP Mutant Grown in Batch Culture*

    PubMed Central

    Thomas, Louise; Hodgson, David A.; Wentzel, Alexander; Nieselt, Kay; Ellingsen, Trond E.; Moore, Jonathan; Morrissey, Edward R.; Legaie, Roxane; Wohlleben, Wolfgang; Rodríguez-García, Antonio; Martín, Juan F.; Burroughs, Nigel J.; Wellington, Elizabeth M. H.; Smith, Margaret C. M.

    2012-01-01

    Bacteria in the genus Streptomyces are soil-dwelling oligotrophs and important producers of secondary metabolites. Previously, we showed that global messenger RNA expression was subject to a series of metabolic and regulatory switches during the lifetime of a fermentor batch culture of Streptomyces coelicolor M145. Here we analyze the proteome from eight time points from the same fermentor culture and, because phosphate availability is an important regulator of secondary metabolite production, compare this to the proteome of a similar time course from an S. coelicolor mutant, INB201 (ΔphoP), defective in the control of phosphate utilization. The proteomes provide a detailed view of enzymes involved in central carbon and nitrogen metabolism. Trends in protein expression over the time courses were deduced from a protein abundance index, which also revealed the importance of stress pathway proteins in both cultures. As expected, the ΔphoP mutant was deficient in expression of PhoP-dependent genes, and several putatively compensatory metabolic and regulatory pathways for phosphate scavenging were detected. Notably there is a succession of switches that coordinately induce the production of enzymes for five different secondary metabolite biosynthesis pathways over the course of the batch cultures. PMID:22147733

  5. S-adenosyl-L-methionine activates actinorhodin biosynthesis by increasing autophosphorylation of the Ser/Thr protein kinase AfsK in Streptomyces coelicolor A3(2).

    PubMed

    Jin, Ying-Yu; Cheng, Jinhua; Yang, Seung Hwan; Meng, Lingzhu; Palaniyandi, Sasikumar Arunachalam; Zhao, Xin-Qing; Suh, Joo-Won

    2011-01-01

    S-Adenosyl-L-methionine (SAM) is one of the major methyl donors in all living organisms. The exogenous treatment with SAM leads to increased actinorhodin production in Streptomyces coelicolor A3(2). In this study, mutants from different stages of the AfsK-AfsR signal transduction cascade were used to test the possible target of SAM. SAM had no significant effect on actinorhodin production in afsK, afsR, afsS, or actII-open reading frame 4 (ORF4) mutant. This confirms that afsK plays a critical role in delivering the signal generated by exogenous SAM. The afsK-pHJL-KN mutant did not respond to SAM, suggesting the involvement of the C-terminal of AfsK in binding with SAM. SAM increased the in vitro autophosphorylation of kinase AfsK in a dose-dependent manner, and also abolished the effect of decreased actinorhodin production by a Ser/Thr kinase inhibitor, K252a. In sum, our results suggest that SAM activates actinorhodin biosynthesis in S. coelicolor M130 by increasing the phosphorylation of protein kinase AfsK.

  6. Epigenetic Activation of Antibacterial Property of an Endophytic Streptomyces coelicolor Strain AZRA 37 and Identification of the Induced Protein Using MALDI TOF MS/MS

    PubMed Central

    Kumar, Jitendra; Sharma, Vijay K.; Singh, Dheeraj K.; Mishra, Ashish; Gond, Surendra K.; Verma, Satish K.; Kumar, Anuj; Kharwar, Ravindra Nath

    2016-01-01

    The endophytic Streptomyces coelicolor strain AZRA 37 was isolated from the surface sterilized root of Azadirachta indica A. Juss., commonly known as neem plant in India. Since only a few reports are available regarding epigenetic modulations of microbial entities, S. coelicolor was treated with different concentrations of 5-azacytidine for this purpose and evaluated for its antibacterial potential against five human pathogenic bacteria (Aeromonas hydrophila IMS/GN11, Enterococcus faecalis IMS/GN7, Salmonella typhi MTCC 3216, Shigella flexneri ATCC 12022 and Staphylococcus aureus ATCC 25923). The crude extract obtained from cultures treated with 25 μM concentration of 5-azacytidine, was found effective against all five pathogenic bacteria tested while the untreated control was only active against 3 pathogenic bacteria. HPLC analysis of crude compounds from treated cultures showed a greater number of compounds than that of the control. Extraction of whole cell protein and its SDS PAGE analysis showed an additional major protein band in 25 μM 5-azacytidine treated culture and MALDI TOF MS/MS analysis revealed that this protein belongs to the porin family. PMID:26844762

  7. Histological Analyses Demonstrate the Temporary Contribution of Yolk Sac, Liver, and Bone Marrow to Hematopoiesis during Chicken Development

    PubMed Central

    Guedes, Priscila Tavares; de Oliveira, Barbara Cristina Euzébio Pereira Dias; Manso, Pedro Paulo de Abreu; Caputo, Luzia Fátima Gonçalves; Pelajo-Machado, Marcelo

    2014-01-01

    The use of avian animal models has contributed to the understanding of many aspects of the ontogeny of the hematopoietic system in vertebrates. However, specific events that occur in the model itself are still unclear. There is a lack of consensus, among previous studies, about which is the intermediate site responsible for expansion and differentiation of hematopoietic cells, and the liver's contribution to the development of this system. Here we aimed to evaluate the presence of hematopoiesis in the yolk sac and liver in chickens, from the stages of intra-aortic clusters in the aorta-genital ridges-mesonephros (AGM) region until hatching, and how it relates to the establishment of the bone marrow. Gallus gallus domesticus L. embryos and their respective yolk sacs at embryonic day 3 (E3) and up to E21 were collected and processed according to standard histological techniques for paraffin embedding. The slides were stained with hematoxylin-eosin, Lennert's Giemsa, and Sirius Red at pH 10.2, and investigated by light microscopy. This study demonstrated that the yolk sac was a unique hematopoietic site between E4 and E12. Hematopoiesis occurred in the yolk sac and bone marrow between E13 and E20. The liver showed granulocytic differentiation in the connective tissue of portal spaces at E15 and onwards. The yolk sac showed expansion of erythrocytic and granulocytic lineages from E6 to E19, and E7 to E20, respectively. The results suggest that the yolk sac is the major intermediate erythropoietic and granulopoietic site where expansion and differentiation occur during chicken development. The hepatic hematopoiesis is restricted to the portal spaces and represented by the granulocytic lineage. PMID:24621665

  8. 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.

  9. 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)

  10. 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)

  11. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1987-01-01

    Describes two demonstrations suitable for chemistry instruction. One involves fractal structures obtained by electrodeposition of silver at an air-water interface and the other deals with molecular weights and music. (TW)

  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

    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)

  14. 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)

  15. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1979-01-01

    Two demonstrations are presented: a verification of the discontinuity of matter based on the law of definite proportions, and a series of consecutive chemical reactions featuring reversible equilibria. (BB)

  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.

    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)

  18. 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)

  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. 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.

  1. Immortalized Mouse Inner Ear Cell Lines Demonstrate a Role for Chemokines in Promoting the Growth of Developing Statoacoustic Ganglion Neurons

    PubMed Central

    Daruwalla, Zeeba; Roth, Therese M.; Attia, Naweah P.; Lukacs, Nicholas W.; Richards, Ayo-Lynn; White, Ian O.; Allen, Susan J.; Barald, Kate F.

    2005-01-01

    The target-derived factors necessary for promoting initial outgrowth from the statoacoustic ganglion (SAG) to the inner ear have not been fully characterized. In the present study, conditioned medium from embryonic Immortomouse inner ear cell lines that maintain many characteristics of developing inner ear sensory epithelia were screened for neurite-promoting activity. Conditioned medium found to be positive for promoting SAG neurite outgrowth and neuronal survival was then tested for the presence of chemokines, molecules that have not previously been investigated for promoting SAG outgrowth. One candidate molecule, monocyte chemotactic protein 1 (MCP-1), was detected in the conditioned medium and subsequently localized to mouse hair cells by immunocytochemistry. In vitro studies demonstrated that function-blocking MCP-1 antibodies decreased the amount of SAG neurite outgrowth induced by the conditioned medium and that subsequent addition of MCP-1 protein was able to promote outgrowth when added to the antibody-treated conditioned medium. The use of the Immortomouse cell lines proved valuable in identifying this candidate cofactor that promotes outgrowth of early-stage SAG nerve fibers and is expressed in embryonic hair cells. PMID:16240240

  2. 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.

  3. 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.

  4. 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.

  5. In Situ Uranium Stabilization through Polyphosphate Remediation: Development and Demonstration at the Hanford Site 300 Area, Washington State

    SciTech Connect

    Wellman, Dawn M.; Pierce, Eric M.; Vermeul, Vincent R.; Mattigod, Shas V.; Richards, Emily L.; Williams, Mark D.; Fruchter, Jonathan S.; Icenhower, Jonathan P.

    2008-06-27

    A site specific treatability test was conducted to optimize polyphosphate remediation technology for implementation through a field-scale technology demonstration to accelerate monitored natural attenuation of the uranium plume within the Hanford 300 Area aquifer. A focused application of polyphosphate was conducted in a source or “hot spot” area to reduce the inventory of available uranium that contributes to the groundwater plume through direct precipitation of uranyl-phosphate solids and secondary containment via precipitation of apatite acting as a long-term sorbent for uranium. The general treatability testing approach consisted of initial site characterization and setup, a polyphosphate injection test, and post-treatment performance assessment. Fundamental science studies were conducted with site specific sediment and groundwater to develop an effective remediation scheme for deployment of polyphosphate technology. In addition to remediating a portion of the plume, the data from this test provides valuable information for designing a full-scale remediation of uranium in the aquifer at the 300 Area of the Hanford Site. It will also provide a detailed understanding of the fundamental underpinnings necessary to evaluate the efficacy and potential utilization of polyphosphate technology at other sites with varying geochemical and hydrodynamic conditions.

  6. 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.

  7. 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.

  8. 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

  9. Tested Demonstrations.

    ERIC Educational Resources Information Center

    Gilbert, George L., Ed.

    1987-01-01

    Describes two demonstrations for use in chemistry instruction. The first illustrates the preparation of a less common oxide of iron, showing why this oxide is rare. The second is an explosion reaction of hydrogen and oxygen that is recommended for use as an attention-getting device. (TW)

  10. 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)

  11. 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)

  12. 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)

  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.

    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…

  16. 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.…

  17. 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)

  18. 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/ .

  19. 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...

  20. cmdABCDEF, a cluster of genes encoding membrane proteins for differentiation and antibiotic production in Streptomyces coelicolor A3(2)

    PubMed Central

    2009-01-01

    Background Streptomyces coelicolor is the most studied Streptomyces species and an excellent model for studying differentiation and antibiotic production. To date, many genes have been identified to be required for its differentiation (e.g. bld genes for aerial growth and whi genes for sporulation) and antibiotics production (including actII-orf4, redD, cdaR as pathway-specific regulatory genes and afsR, absA1/A2 as pleiotropic regulatory genes). Results A gene cluster containing six genes (SCO4126-4131) was proved to be co-transcribed in S. coelicolor. Deletions of cmdABCDEF (SCO4126-4131) displayed defective sporulation including formation of aberrant branches, and abnormalities in chromosome segregation and spore septation. Disruption mutants of apparently orthologous genes of S. lividans and S. avermitilis also showed defective sporulation, implying that the role of these genes is similar among Streptomyces. Transcription of cmdB, and therefore presumably of the whole operon, was regulated developmentally. Five of the encoded proteins (CmdA, C, D, E, F) were predicted membrane proteins. The other, CmdB, a predicted ATP/GTP-binding protein with an ABC-transporter-ATPase domain shown here to be essential for its function, was also located on the cell membrane. These results indicate that CmdABCDEF proteins mainly affect Streptomyces differentiation at an early stage of aerial hyphae formation, and suggest that these proteins may form a complex on cell membrane for proper segregation of chromosomes. In addition, deletions of cmdABCDEF also revealed over-production of blue-pigmented actinorhodin (Act) via activation of transcription of the pathway-specific regulatory gene actII-orf4 of actinorhodin biosynthesis. Conclusion In this study, six co-transcribed genes cmdABCDEF were identified by their effects on differentiation and antibiotic production in Streptomyces coelicolor A3(2). These six membrane-located proteins are possibly assembled into a complex to

  1. Tomographic data developed using the ABEM RAMAC borehole radar system at the Mixed Waste Landfill Integrated Demonstration

    SciTech Connect

    MacLeod, G.A.; Barker, D.L.; Molnar, S.

    1994-02-18

    The ABEM RAMAC borehole radar system was run as part of the Mixed Waste Landfill Integrated Demonstration for Sandia National Laboratories at Kirtland AFB. Tomograms were created between three test boreholes-UCAP No. 1, UCAP No. 2, and UCAP No. 3. These tomograms clearly delineate areas of amplitude attenuation and residual time of arrival or slowness differences. Plots for slowness were made using both the maximum and minimum of the first arrival pulse. The data demonstrates that the ABEM RAMAC 60-MHz pulse sampling radar system can be used to collect usable data in a highly conductive environment.

  2. A National Plan for Energy Research, Development & Demonstration: Creating Energy Choices for the Future, 1976. Volume 1: The Plan.

    ERIC Educational Resources Information Center

    Energy Research and Development Administration, Washington, DC.

    This report presents a plan developed by the Energy Research and Development Administration (ERDA) concerning future energy sources. The plan summarizes ERDA's current views on the energy technologies the U.S. will need to achieve long-term energy independence. The role of the private sector in the development and commercialization of new energy…

  3. 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…

  4. 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…

  5. Project to develop and demonstrate an inexpensive automatic drop mechanism for window insulation systems. Final progress report

    SciTech Connect

    Not Available

    1985-01-01

    The EPR concept is technically practical, but lacks something which is crucial to its marketplace success. That ''something'' would appear to be the capability of fully automatic operation, although logic says that semi-automatic operation obtains the benefits of window insulation closure. That fully-automatic operation produces a vastly different response in the marketplace is indicated by the fact that over 35 demonstrators at $30 each have been ordered in little more than one month since the Automatic Shade Roller System has been offered for sale; whereas, with nearly 1000 inquiries responded to, only a dozen EPR's were sold at $12.50. The role of automatic rollers in window insulation systems has not been thoroughly demonstrated. They can be used to operate exterior sunscreen shades, between-glazing shades, and combinations of sunscreen and insulating shades. What would be appropriate at this time would be installation of a combination of automatic shading/insulating systems on the windows of a public building or home to demonstrate the energy-savings benefits of such installations. As the sole US manufacturer of automatic shade rollers, Solar Roller Corporation, an associate with Solar Power West, is the ideal organization to make such installations.

  6. Structural and genomic DNA analysis of the putative TetR transcriptional repressor SCO7518 from Streptomyces coelicolor A3(2).

    PubMed

    Hayashi, Takeshi; Tanaka, Yoshikazu; Sakai, Naoki; Okada, Ui; Yao, Min; Watanabe, Nobuhisa; Tamura, Tomohiro; Tanaka, Isao

    2014-11-28

    SCO7518 is a protein of unknown function from Streptomyces coelicolor A3(2) that has been classified into the TetR transcriptional regulator family. In this study, a crystal structure of SCO7518 was determined at 2.29Å resolution. The structure is a homodimer of protomers that comprise an N-terminal DNA-binding domain and a C-terminal dimerization and regulatory domain, and possess a putative ligand-binding cavity. Genomic systematic evolution of ligands by exponential enrichment and electrophoretic mobility shift assays revealed that SCO7518 specifically binds to an operator sequence located upstream of the sco7519 gene, which encodes a maltose O-acetyltransferase. These results suggest that SCO7518 is a transcriptional repressor of sco7519 expression.

  7. 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)

  8. 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)

  9. 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...

  10. TrpM, a Small Protein Modulating Tryptophan Biosynthesis and Morpho-Physiological Differentiation in Streptomyces coelicolor A3(2)

    PubMed Central

    Palazzotto, Emilia; Gallo, Giuseppe; Renzone, Giovanni; Giardina, Anna; Sutera, Alberto; Silva, Joohee; Vocat, Celinè; Botta, Luigi; Scaloni, Andrea; Puglia, Anna Maria

    2016-01-01

    In the model actinomycete Streptomyces coelicolor A3(2), small open reading frames encoding proteins with unknown functions were identified in several amino acid biosynthetic gene operons, such as SCO2038 (trpX) in the tryptophan trpCXBA locus. In this study, the role of the corresponding protein in tryptophan biosynthesis was investigated by combining phenotypic and molecular analyses. The 2038KO mutant strain was characterized by delayed growth, smaller aerial hyphae and reduced production of spores and actinorhodin antibiotic, with respect to the WT strain. The capability of this mutant to grow on minimal medium was rescued by tryptophan and tryptophan precursor (serine and/or indole) supplementation on minimal medium and by gene complementation, revealing the essential role of this protein, here named TrpM, as modulator of tryptophan biosynthesis. His-tag pull-down and bacterial adenylate cyclase-based two hybrid assays revealed TrpM interaction with a putative leucyl-aminopeptidase (PepA), highly conserved component among various Streptomyces spp. In silico analyses showed that PepA is involved in the metabolism of serine, glycine and cysteine through a network including GlyA, CysK and CysM enzymes. Proteomic experiments suggested a TrpM-dependent regulation of metabolic pathways and cellular processes that includes enzymes such as GlyA, which is required for the biosynthesis of tryptophan precursors and key proteins participating in the morpho-physiological differentiation program. Altogether, these findings reveal that TrpM controls tryptophan biosynthesis at the level of direct precursor availability and, therefore, it is able to exert a crucial effect on the morpho-physiological differentiation program in S. coelicolor A3(2). PMID:27669158

  11. The bldD Gene of Streptomyces coelicolor A3(2): a Regulatory Gene Involved in Morphogenesis and Antibiotic Production

    PubMed Central

    Elliot, Marie; Damji, Farzana; Passantino, Rosa; Chater, Keith; Leskiw, Brenda

    1998-01-01

    The bld mutants of Streptomyces coelicolor A3(2) are blocked at the earliest stage of sporulation, the formation of aerial hyphae, and are pleiotropically defective in antibiotic production. Using a phage library of wild-type S. coelicolor DNA, we isolated a recombinant phage which restored both sporulation and antibiotic production to strains carrying the single known bldD mutation. Nucleotide sequence analysis of a 1.3-kb complementing subclone identified an open reading frame, designated bldD, encoding a translation product of 167 amino acid residues. Nucleotide sequence analysis of the bldD-containing fragment amplified from the chromosome of a bldD mutant strain revealed a point mutation changing a tyrosine residue at amino acid position 62 to a cysteine. Although a comparison of the BldD sequence to known proteins in the databases failed to show any strong similarities, analysis of the BldD sequence for secondary structural elements did reveal a putative helix-turn-helix, DNA recognition element near the C terminus of the protein. A comparison of bldD transcript levels in the bldD+ and bldD mutant strains using both Northern blot analysis and S1 nuclease protection studies showed vast overexpression of bldD transcripts in the mutant, suggesting that BldD negatively regulates its own synthesis. High-resolution S1 nuclease mapping identified the transcription start point as a G residue 63 nucleotides upstream from the bldD start codon and 7 nucleotides downstream from −10 and −35 sequences resembling E. coli-like streptomycete promoters. PMID:9515925

  12. 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.

  13. The development and demonstration of hybrid programmable attitude control electronics. [with adaptable analog/digital design approach

    NASA Technical Reports Server (NTRS)

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

    1974-01-01

    HYPACE provides an adaptable, analog/digital design approach that permits preflight and in-flight accommodation of mission changes, component performance variations, spacecraft changes, etc., through programing. This enabled broad multimission flexibility of application in a cost-effective manner. 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. The success of this effort resulted in the concept being selected for the Mariner/Jupiter/Saturn 1977 spacecraft application.

  14. Development and demonstration of manufacturing processes for fabricating graphite/Larc-160 polyimide structural elements, part 4, paragraph B

    NASA Technical Reports Server (NTRS)

    1981-01-01

    Progress in the development of processes for production of Celion/LARC-160 graphite-polyimide materials, quality control, and the fabrication of Space Shuttle composite structure components is reported. Liquid chromatographic analyses of three repeatibility batches were performed and are compared to previous Hexcel standard production and to variables study LARC-160 intermediate resins. Development of processes for chopped fiber molding are described and flexural strength, elastic modulus, and other physical and mechanical properties of the molding are presented.

  15. Strategy for developing environmental sampling and analysis programs at coal-gasification pilot and demonstration plants. Topical report

    SciTech Connect

    Skinner, F.D.

    1989-02-01

    This report provides assistance in the development of sampling and analysis programs to obtain the data needed to support environmental permitting and environmental control-system design and evaluation for coal-to-substitute natural gas plants. The approach involved in the development of such a program involves: (1) determining the emission and discharge streams from the plant; (2) deciding which of the species that are likely to be present in these streams will be controlled under major Federal environmental regulations or need to be measured to evaluate the performance of potential environmental control processes; and (3) developing a test plant for obtaining the required data. A step-by-step example is given illustrating how to use the information presented in the report.

  16. AIST-NREL Concentrator Photovoltaic (CPV) Demonstration. Cooperative Research and Development Final Report, CRADA Number CRD-10-402

    SciTech Connect

    Kurtz, Sarah

    2015-05-11

    The purpose of the project is to demonstrate and quantitatively compare performance of CPV systems installed in Japan and in the United States. The deployment conditions (e.g. spectrum and temperature) are site dependent and the optimal design of the system may vary with location. The CPV systems will use multi-junction concentrator cells for the conversion of sunlight into electricity. The optimal design of the cell may depend on the location at which a CPV system is installed. Thus, the systems in Japan and in the U.S. will all use a combination of concentrator cells obtained from three different vendors. This CRADA pertains only to the equipment that will be installed in the U.S. This effort is a collaborative project between AIST and NREL.

  17. Development of DANDYs, new 3,5-diaryl-7-azaindoles demonstrating potent DYRK1A kinase inhibitory activity.

    PubMed

    Gourdain, Stéphanie; Dairou, Julien; Denhez, Clément; Bui, Linh Chi; Rodrigues-Lima, Fernando; Janel, Nathalie; Delabar, Jean M; Cariou, Kevin; Dodd, Robert H

    2013-12-12

    A series of 3,5-diaryl-1H-pyrrolo[2,3-b]pyridines were synthesized and evaluated for inhibition of DYRKIA kinase in vitro. Derivatives having hydroxy groups on the aryl moieties (2c, 2j-l) demonstrated high inhibitory potencies with Kis in the low nanomolar range. Their methoxy analogues were up to 100 times less active. Docking studies at the ATP binding site suggested that these compounds bind tightly to this site via a network of multiple H-bonds with the peptide backbone. None of the active compounds were cytotoxic to KB cells at 10(-6) M. Kinase profiling revealed that compound 2j showed 2-fold selectivity for DYRK1A with respect to DYRK2 and DYRK3. PMID:24188002

  18. Develop and Demonstrate the Cellulose to Ethanol Process: Executive Summary of the Final Technical Report, 17 September 1980 - 17 March 1982

    SciTech Connect

    Emert, George H.; Becker, Dana K.; Bevernitz, Kurt J.; Gracheck, Stephen J.; Kienholz, Eldon W.; Rivers, Dougals B.; Zoldak, Bernadette R.; Woodford, Lindley C.

    1982-01-01

    The Biomass Research Center at the University of Arkansas was contracted by the Solar Energy Research Institute to 'Develop and Demonstrate the Cellulose to Ethanol Process.' The purpose of the contract was to accelerate site selection, site specific engineering, and research and development leading to the determination of the feasibility of economically operating a cellulose to ethanol commercial scale plant.

  19. Development and Demonstration of Material Properties Database and Software for the Simulation of Flow Properties in Cementitious Materials

    SciTech Connect

    Smith, F.; Flach, G.

    2015-03-30

    This report describes work performed by the Savannah River National Laboratory (SRNL) in fiscal year 2014 to develop a new Cementitious Barriers Project (CBP) software module designated as FLOExcel. FLOExcel incorporates a uniform database to capture material characterization data and a GoldSim model to define flow properties for both intact and fractured cementitious materials and estimate Darcy velocity based on specified hydraulic head gradient and matric tension. The software module includes hydraulic parameters for intact cementitious and granular materials in the database and a standalone GoldSim framework to manipulate the data. The database will be updated with new data as it comes available. The software module will later be integrated into the next release of the CBP Toolbox, Version 3.0. This report documents the development efforts for this software module. The FY14 activities described in this report focused on the following two items that form the FLOExcel package; 1) Development of a uniform database to capture CBP data for cementitious materials. In particular, the inclusion and use of hydraulic properties of the materials are emphasized; and 2) Development of algorithms and a GoldSim User Interface to calculate hydraulic flow properties of degraded and fractured cementitious materials. Hydraulic properties are required in a simulation of flow through cementitious materials such as Saltstone, waste tank fill grout, and concrete barriers. At SRNL these simulations have been performed using the PORFLOW code as part of Performance Assessments for salt waste disposal and waste tank closure.

  20. Develop and Demonstrate the Use of Integrated Instructional Delivery Systems for Application in Technical/Vocational Education. Final Report.

    ERIC Educational Resources Information Center

    San Antonio State Hospital, TX. Office of Education Services.

    This report presents a discussion of important issues surrounding the use of interactive instructional technology by vocational educators. It considers interactive instructional television system components with special attention to system interface considerations, software considerations, and design and development of interactive instructional…