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Sample records for ancillary pilin subunit

  1. Lactobacillus rhamnosus GG SpaC pilin subunit binds to the carbohydrate moieties of intestinal glycoconjugates.

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    Nishiyama, Keita; Ueno, Shintaro; Sugiyama, Makoto; Yamamoto, Yuji; Mukai, Takao

    2016-06-01

    Lactobacillus rhamnosus GG (LGG) is a well-established probiotic strain. The beneficial properties of this strain are partially dependent on its prolonged residence in the gastrointestinal tract, and are likely influenced by its adhesion to the intestinal mucosa. The pilin SpaC subunit, located within the Spa pili structure, is the most well studied LGG adhesion factor. However, the binding epitopes of SpaC remain largely unknown. The aim of this study was to evaluate the binding properties of SpaC to the carbohydrate moieties of intestinal glycoconjugates using a recombinant SpaC protein. In a competitive enzyme-linked immunosorbent assay, SpaC binding was markedly reduced by addition of purified mucin and the mucin oligosaccharide fraction. Histochemical staining revealed that the binding of SpaC was drastically reduced by periodic acid treatment. Moreover, in the surface plasmon resonance-based Biacore assay, SpaC bound strongly to the carbohydrate moieties containing β-galactoside at the non-reducing terminus of glycolipids. We here provide the first demonstration that SpaC binds to the oligosaccharide chains of mucins, and that the carbohydrate moieties containing β-galactoside at the non-reducing termini of glycoconjugates play a crucial role in this binding. Our results demonstrate the importance of carbohydrates of SpaC for mucus interactions. © 2015 Japanese Society of Animal Science.

  2. New insights about pilus formation in gut-adapted Lactobacillus rhamnosus GG from the crystal structure of the SpaA backbone-pilin subunit

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    Chaurasia, Priyanka; Pratap, Shivendra; von Ossowski, Ingemar; Palva, Airi; Krishnan, Vengadesan

    2016-01-01

    Thus far, all solved structures of pilin-proteins comprising sortase-assembled pili are from pathogenic genera and species. Here, we present the first crystal structure of a pilin subunit (SpaA) from a non-pathogen host (Lactobacillus rhamnosus GG). SpaA consists of two tandem CnaB-type domains, each with an isopeptide bond and E-box motif. Intriguingly, while the isopeptide bond in the N-terminal domain forms between lysine and asparagine, the one in the C-terminal domain atypically involves aspartate. We also solved crystal structures of mutant proteins where residues implicated in forming isopeptide bonds were replaced. Expectedly, the E-box-substituted E139A mutant lacks an isopeptide bond in the N-terminal domain. However, the C-terminal E269A substitution gave two structures; one of both domains with their isopeptide bonds present, and another of only the N-terminal domain, but with an unformed isopeptide bond and significant conformational changes. This latter crystal structure has never been observed for any other Gram-positive pilin. Notably, the C-terminal isopeptide bond still forms in D295N-substituted SpaA, irrespective of E269 being present or absent. Although E-box mutations affect SpaA proteolytic and thermal stability, a cumulative effect perturbing normal pilus polymerization was unobserved. A model showing the polymerized arrangement of SpaA within the SpaCBA pilus is proposed. PMID:27349405

  3. Cloning, expression, purification, crystallization and X-ray crystallographic analysis of CofB, the minor pilin subunit of CFA/III from human enterotoxigenic Escherichia coli.

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    Kawahara, Kazuki; Oki, Hiroya; Fukakusa, Shunsuke; Maruno, Takahiro; Kobayashi, Yuji; Motooka, Daisuke; Taniguchi, Tooru; Honda, Takeshi; Iida, Tetsuya; Nakamura, Shota; Ohkubo, Tadayasu

    2015-06-01

    Colonization factor antigen III (CFA/III) is one of the virulence factors of human enterotoxigenic Escherichia coli (ETEC) that forms the long, thin, proteinaceous fibres of type IV pili through assembly of its major and minor subunits CofA and CofB, respectively. The crystal structure of CofA has recently been reported; however, the lack of structural information for CofB, the largest among the known type IV pilin subunits, hampers a comprehensive understanding of CFA/III pili. In this study, constructs of wild-type CofB with an N-terminal truncation and the corresponding SeMet derivative were cloned, expressed, purified and crystallized. The crystals belonged to the rhombohedral space group R32, with unit-cell parameters a = b = 103.97, c = 364.57 Å for the wild-type construct and a = b = 103.47, c = 362.08 Å for the SeMet-derivatized form. Although the diffraction quality of these crystals was initially very poor, dehydration of the crystals substantially improved the resolution limit from ∼ 4.0 to ∼ 2.0 Å. The initial phase was solved by the single-wavelength anomalous dispersion (SAD) method using a dehydrated SeMet CofB crystal, which resulted in an interpretable electron-density map.

  4. Structure of the CFA/III major pilin subunit CofA from human enterotoxigenic Escherichia coli determined at 0.90 Å resolution by sulfur-SAD phasing.

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    Fukakusa, Shunsuke; Kawahara, Kazuki; Nakamura, Shota; Iwashita, Takaki; Baba, Seiki; Nishimura, Mitsuhiro; Kobayashi, Yuji; Honda, Takeshi; Iida, Tetsuya; Taniguchi, Tooru; Ohkubo, Tadayasu

    2012-10-01

    CofA, a major pilin subunit of colonization factor antigen III (CFA/III), forms pili that mediate small-intestinal colonization by enterotoxigenic Escherichia coli (ETEC). In this study, the crystal structure of an N-terminally truncated version of CofA was determined by single-wavelength anomalous diffraction (SAD) phasing using five sulfurs in the protein. Given the counterbalance between anomalous signal strength and the undesired X-ray absorption of the solvent, diffraction data were collected at 1.5 Å resolution using synchrotron radiation. These data were sufficient to elucidate the sulfur substructure at 1.38 Å resolution. The low solvent content (29%) of the crystal necessitated that density modification be performed with an additional 0.9 Å resolution data set to reduce the phase error caused by the small sulfur anomalous signal. The CofA structure showed the αβ-fold typical of type IVb pilins and showed high structural homology to that of TcpA for toxin-coregulated pili of Vibrio cholerae, including spatial distribution of key residues critical for pilin self-assembly. A pilus-filament model of CofA was built by computational docking and molecular-dynamics simulation using the previously reported filament model of TcpA as a structural template. This model revealed that the CofA filament surface was highly negatively charged and that a 23-residue-long loop between the α1 and α2 helices filled the gap between the pilin subunits. These characteristics could provide a unique binding epitope for the CFA/III pili of ETEC compared with other type IVb pili.

  5. Pilin Processing Follows a Different Temporal Route than That of Archaellins in Methanococcus maripaludis

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    Divya B. Nair

    2015-01-01

    Full Text Available Methanococcus maripaludis has two different surface appendages: type IV-like pili and archaella. Both structures are believed to be assembled using a bacterial type IV pilus mechanism. Each structure is composed of multiple subunits, either pilins or archaellins. Both pilins and archaellins are made initially as preproteins with type IV pilin-like signal peptides, which must be removed by a prepilin peptidase-like enzyme. This enzyme is FlaK for archaellins and EppA for pilins. In addition, both pilins and archaellins are modified with N-linked glycans. The archaellins possess an N-linked tetrasaccharide while the pilins have a pentasaccharide which consists of the archaellin tetrasaccharide but with an additional sugar, an unidentified hexose, attached to the linking sugar. In this report, we show that archaellins can be processed by FlaK in the absence of N-glycosylation and N-glycosylation can occur on archaellins that still retain their signal peptides. In contrast, pilins are not glycosylated unless they have been acted on by EppA to have the signal peptide removed. However, EppA can still remove signal peptides from non-glycosylated pilins. These findings indicate that there is a difference in the order of the posttranslational modifications of pilins and archaellins even though both are type IV pilin-like proteins.

  6. Purification, crystallization and preliminary X-ray diffraction analysis of SpaD, a backbone-pilin subunit encoded by the fimbrial spaFED operon in Lactobacillus rhamnosus GG.

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    Chaurasia, Priyanka; von Ossowski, Ingemar; Palva, Airi; Krishnan, Vengadesan

    2015-01-01

    SpaD is the predicted backbone-pilin subunit of the SpaFED pilus, whose loci are encoded by the fimbrial spaFED operon in Lactobacillus rhamnosus GG, a Gram-positive gut-adapted commensal strain with perceived probiotic benefits. In this study, soluble recombinant SpaD protein was overproduced in Escherichia coli and then purified by Ni2+-chelating affinity and gel-filtration chromatography. After limited proteolysis with α-chymotrypsin, good-quality crystals of SpaD were obtained which diffracted beyond 2.0 Å resolution. These crystals belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a=50.11, b=83.27, c=149.65 Å. For phasing, sodium iodide-derivatized crystals were prepared using the halide quick-soaking method and diffraction data were collected in-house to a resolution of 2.2 Å. An interpretable electron-density map was successfully obtained using single-wavelength anomalous diffraction (SAD).

  7. Crystallization and preliminary X-ray diffraction analysis of PsaA, the adhesive pilin subunit that forms the pH 6 antigen on the surface of Yersinia pestis

    International Nuclear Information System (INIS)

    Bao, Rui; Esser, Lothar; Sadhukhan, Annapurna; Nair, Manoj K. M.; Schifferli, Dieter M.; Xia, Di

    2012-01-01

    The pH 6 antigen Psa displayed on the surface of Yersinia pestis, the bacterium that causes plague in humans, consists of polymers of a single protein subunit termed PsaA. Donor-strand complemented PsaA was purified and crystallized. Yersinia pestis has been responsible for a number of high-mortality epidemics throughout human history. Like all other bacterial infections, the pathogenesis of Y. pestis begins with the attachment of bacteria to the surface of host cells. At least five surface proteins from Y. pestis have been shown to interact with host cells. Psa, the pH 6 antigen, is one of them and is deployed on the surface of bacteria as thin flexible fibrils that are the result of the polymerization of a single PsaA pilin subunit. Here, the crystallization of recombinant donor-strand complemented PsaA by the hanging-drop vapor-diffusion method is reported. X-ray diffraction data sets were collected to 1.9 Å resolution from a native crystal and to 1.5 Å resolution from a bromide-derivatized crystal. These crystals displayed the symmetry of the orthorhombic space group P222 1 , with unit-cell parameters a = 26.3, b = 54.6, c = 102.1 Å. Initial phases were derived from single isomorphous replacement with anomalous scattering experiments, resulting in an electron-density map that showed a single molecule in the crystallographic asymmetric unit. Sequence assignment was aided by residues binding to bromide ions of the heavy-atom derivative

  8. Intron retention in mRNA encoding ancillary subunit of insect voltage-gated sodium channel modulates channel expression, gating regulation and drug sensitivity.

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    Céline M Bourdin

    Full Text Available Insect voltage-gated sodium (Nav channels are formed by a well-known pore-forming α-subunit encoded by para-like gene and ancillary subunits related to TipE from the mutation "temperature-induced-paralysis locus E." The role of these ancillary subunits in the modulation of biophysical and pharmacological properties of Na(+ currents are not enough documented. The unique neuronal ancillary subunit TipE-homologous protein 1 of Drosophila melanogaster (DmTEH1 strongly enhances the expression of insect Nav channels when heterologously expressed in Xenopus oocytes. Here we report the cloning and functional expression of two neuronal DmTEH1-homologs of the cockroach, Periplaneta americana, PaTEH1A and PaTEH1B, encoded by a single bicistronic gene. In PaTEH1B, the second exon encoding the last 11-amino-acid residues of PaTEH1A is shifted to 3'UTR by the retention of a 96-bp intron-containing coding-message, thus generating a new C-terminal end. We investigated the gating and pharmacological properties of the Drosophila Nav channel variant (DmNav1-1 co-expressed with DmTEH1, PaTEH1A, PaTEH1B or a truncated mutant PaTEH1Δ(270-280 in Xenopus oocytes. PaTEH1B caused a 2.2-fold current density decrease, concomitant with an equivalent α-subunit incorporation decrease in the plasma membrane, compared to PaTEH1A and PaTEH1Δ(270-280. PaTEH1B positively shifted the voltage-dependences of activation and slow inactivation of DmNav1-1 channels to more positive potentials compared to PaTEH1A, suggesting that the C-terminal end of both proteins may influence the function of the voltage-sensor and the pore of Nav channel. Interestingly, our findings showed that the sensitivity of DmNav1-1 channels to lidocaine and to the pyrazoline-type insecticide metabolite DCJW depends on associated TEH1-like subunits. In conclusion, our work demonstrates for the first time that density, gating and pharmacological properties of Nav channels expressed in Xenopus oocytes can be

  9. Crystal Structure of the Minor Pilin CofB, the Initiator of CFA/III Pilus Assembly in Enterotoxigenic Escherichia coli.

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    Kolappan, Subramania; Ng, Dixon; Yang, Guixiang; Harn, Tony; Craig, Lisa

    2015-10-23

    Type IV pili are extracellular polymers of the major pilin subunit. These subunits are held together in the pilus filament by hydrophobic interactions among their N-terminal α-helices, which also anchor the pilin subunits in the inner membrane prior to pilus assembly. Type IV pilus assembly involves a conserved group of proteins that span the envelope of Gram-negative bacteria. Among these is a set of minor pilins, so named because they share their hydrophobic N-terminal polymerization/membrane anchor segment with the major pilins but are much less abundant. Minor pilins influence pilus assembly and retraction, but their precise functions are not well defined. The Type IV pilus systems of enterotoxigenic Escherichia coli and Vibrio cholerae are among the simplest of Type IV pilus systems and possess only a single minor pilin. Here we show that the enterotoxigenic E. coli minor pilins CofB and LngB are required for assembly of their respective Type IV pili, CFA/III and Longus. Low levels of the minor pilins are optimal for pilus assembly, and CofB can be detected in the pilus fraction. We solved the 2.0 Å crystal structure of N-terminally truncated CofB, revealing a pilin-like protein with an extended C-terminal region composed of two discrete domains connected by flexible linkers. The C-terminal region is required for CofB to initiate pilus assembly. We propose a model for CofB-initiated pilus assembly with implications for understanding filament growth in more complex Type IV pilus systems as well as the related Type II secretion system. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  10. Homo-trimeric Structure of the Type IVb Minor Pilin CofB Suggests Mechanism of CFA/III Pilus Assembly in Human Enterotoxigenic Escherichia coli.

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    Kawahara, Kazuki; Oki, Hiroya; Fukakusa, Shunsuke; Yoshida, Takuya; Imai, Tomoya; Maruno, Takahiro; Kobayashi, Yuji; Motooka, Daisuke; Iida, Tetsuya; Ohkubo, Tadayasu; Nakamura, Shota

    2016-03-27

    In gram-negative bacteria, the assembly of type IV pilus (T4P) and the evolutionally related pseudopilus of type II secretion system involves specialized structural proteins called pilins and pseudopilins, respectively, and is dynamically regulated to promote bacterial pathogenesis. Previous studies have suggested that a structural "tip"-like hetero-complex formed through the interaction of at least three minor (pseudo) pilins plays an important role in this process, while some members of the pathogenic type IVb subfamily are known to have only one such minor pilin subunit whose function is still unknown. Here, we determined the crystal structure of the type IVb minor pilin CofB of colonization factor antigen/III from human enterotoxigenic Escherichia coli at 1.88-Å resolution. The crystal structure, in conjunction with physicochemical analysis in solution, reveals a symmetrical homo-trimeric arrangement distinct from the hetero-complexes of minor (pseudo) pilins observed in other T4P and type II secretion systems. Each CofB monomer adopts a unique three-domain architecture, in which the C-terminal β-sheet-rich lectin domain can effectively initiate trimer association of its pilin-like N-terminal domain through extensive hydrophobic interactions followed by domain swapping at the central hinge-like domain. Deletion of cofB produces a phenotype with no detectable pili formation on the cell surface, while molecular modeling indicates that the characteristic homo-trimeric structure of CofB is well situated at the pilus tip of colonization factor antigen/III formed by the major pilin CofA, suggesting a role for the minor pilin in the efficient initiation of T4P assembly. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. Structure of the Type IVa Major Pilin from the Electrically Conductive Bacterial Nanowires of Geobacter sulfurreducens

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    Reardon, Patrick N.; Mueller, Karl T.

    2013-10-11

    Several species of bacteria are capable of reducing insoluble metal oxides as well as other extracellular electron acceptors. These bacteria play a critical role in the cycling of minerals in subsurface environments, sediments, and groundwater. In some species of bacteria, such as Geobacter sulfurreducens, the transport of electrons is facilitated by filamentous fibers that are referred to as bacterial nanowires. These nanowires belong to the type IVa family of pilin proteins and are mainly comprised of one subunit protein, PilA. Here, we report the high resolution solution nuclear magnetic resonance (NMR) structure of the PilA protein from G. sulfurreducens determined in detergent micelles. The protein is over 85% α-helical and exhibits similar architecture to the N-terminal regions of other non-conductive type IVa pilins. The detergent micelle interacts with the first 21 amino acids of the protein, indicating that this region likely associates with the bacterial inner membrane prior to fiber formation. A model of the G. sulfurreducens pilus fiber is proposed based on docking of this structure into the fiber model of the type IVa pilin from Neisseria gonorrhoeae. This model provides insight into the organization of aromatic amino acids that are important for electrical conduction.

  12. Expression, purification, crystallization and preliminary crystallographic analysis of SpaA, a major pilin from Corynebacterium diphtheriae

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    Kang, Hae Joo; Paterson, Neil G.; Baker, Edward N.

    2009-01-01

    SpaA, one of the major pilins of C. diphtheriae, has been expressed, purified and crystallized and X-ray diffraction data have been collected to 1.6 Å resolution. Bacterial pili are cell-surface organelles that are critically involved in adhesion to host cells, leading to the colonization of host tissues and the establishment of infections. Whereas the pili of Gram-negative bacteria have been extensively studied, those of Gram-positive bacteria came to light only recently after the discovery and characterization of Corynebacterium diphtheriae pili. These newly discovered pili are formed by the covalent polymerization of pilin subunits catalyzed by sortase enzymes, making them fundamentally different from the noncovalent pilin assemblies of Gram-negative bacteria. Here, the expression, crystallization and preliminary crystallographic analysis of SpaA, which forms the shaft of one of the three types of pili expressed by C. diphtheriae, are reported. SpaA 53–486 crystals diffracted to 1.6 Å resolution and belonged to space group P2 1 2 1 2 1 , with unit-cell parameters a = 34.9, b = 64.1, c = 198.7 Å, α = β = γ = 90°

  13. A Pilin Region Affecting Host Range of the Pseudomonas aeruginosa RNA Phage, PP7

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    Eun Sook Kim

    2018-02-01

    Full Text Available The host range of a phage is determined primarily by phage-receptor interaction. Here, we profiled the host range of an RNA leviphage, PP7 that requires functional type IV pilus (TFP in order to enter into its host bacterium, Pseudomonas aeruginosa. Out of 25 twitching-proficient P. aeruginosa strains, 4 with group I pilin and 7 with group III pilin displayed PP7-resistance. The remaining 14 possessed group II pilin, which included 10 PP7-sensitive and 4 PP7-resistant strains, suggesting that only the strains with TFP consisted of a subset of group II (hence, group IIa pilin were susceptible to PP7. The co-expression of the PAO1 (group IIa pilin rendered all the strains susceptible to PP7, with the exception of the 4 strains with group I pilin. Moreover, the expression of PA14 (group III and PAK (group IIb pilin in the PAO1 pilA mutant restored the twitching motility but not the PP7-suceptibility. Site-directed and random mutation analyses of PAO1 pilin enabled us to identify a pilin mutant (G96S that is fully functional but resistant to PP7 infection. This is due to the lack of any phage-receptor interactions, suggesting the structural properties of the β1-β2 loop in the variable region 2 of the group II pilin might be involved in PP7 infection.

  14. Neisseria meningitidis Type IV Pili Composed of Sequence Invariable Pilins Are Masked by Multisite Glycosylation.

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    Joseph Gault

    2015-09-01

    Full Text Available The ability of pathogens to cause disease depends on their aptitude to escape the immune system. Type IV pili are extracellular filamentous virulence factors composed of pilin monomers and frequently expressed by bacterial pathogens. As such they are major targets for the host immune system. In the human pathogen Neisseria meningitidis, strains expressing class I pilins contain a genetic recombination system that promotes variation of the pilin sequence and is thought to aid immune escape. However, numerous hypervirulent clinical isolates express class II pilins that lack this property. This raises the question of how they evade immunity targeting type IV pili. As glycosylation is a possible source of antigenic variation it was investigated using top-down mass spectrometry to provide the highest molecular precision on the modified proteins. Unlike class I pilins that carry a single glycan, we found that class II pilins display up to 5 glycosylation sites per monomer on the pilus surface. Swapping of pilin class and genetic background shows that the pilin primary structure determines multisite glycosylation while the genetic background determines the nature of the glycans. Absence of glycosylation in class II pilins affects pilus biogenesis or enhances pilus-dependent aggregation in a strain specific fashion highlighting the extensive functional impact of multisite glycosylation. Finally, molecular modeling shows that glycans cover the surface of class II pilins and strongly decrease antibody access to the polypeptide chain. This strongly supports a model where strains expressing class II pilins evade the immune system by changing their sugar structure rather than pilin primary structure. Overall these results show that sequence invariable class II pilins are cloaked in glycans with extensive functional and immunological consequences.

  15. Isopeptide bonds of the major pilin protein BcpA influence pilus structure and bundle formation on the surface of Bacillus cereus

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    Hendrickx, Antoni P.A.; Poor, Catherine B.; Jureller, Justin E.; Budzik, Jonathan M.; He, Chuan; Schneewind, Olaf (UC)

    2012-09-05

    Bacillus cereus strains elaborate pili on their surface using a mechanism of sortase-mediated cross-linking of major and minor pilus components. Here we used a combination of electron microscopy and atomic force microscopy to visualize these structures. Pili occur as single, double or higher order assemblies of filaments formed from monomers of the major pilin, BcpA, capped by the minor pilin, BcpB. Previous studies demonstrated that within assembled pili, four domains of BcpA -- CNA{sub 1}, CNA{sub 2}, XNA and CNA{sub 3} -- each acquire intramolecular lysine-asparagine isopeptide bonds formed via catalytic glutamic acid or aspartic acid residues. Here we showed that mutants unable to form the intramolecular isopeptide bonds in the CNA2 or CNA3 domains retain the ability to form pilus bundles. A mutant lacking the CNA{sub 1} isopeptide bond assembled deformed pilin subunits that failed to associate as bundles. X-ray crystallography revealed that the BcpA variant Asp{sup 312}Ala, lacking an aspartyl catalyst, did not generate the isopeptide bond within the jelly-roll structure of XNA. The Asp{sup 312}Ala mutant was also unable to form bundles and promoted the assembly of deformed pili. Thus, structural integrity of the CNA{sub 1} and XNA domains are determinants for the association of pili into higher order bundle structures and determine native pilus structure.

  16. A slow-forming isopeptide bond in the structure of the major pilin SpaD from Corynebacterium diphtheriae has implications for pilus assembly

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    Kang, Hae Joo; Paterson, Neil G. [University of Auckland, Private Bag 92019, Auckland 1142 (New Zealand); Kim, Chae Un [Cornell University, Ithaca, NY 14853 (United States); Middleditch, Martin [University of Auckland, Private Bag 92019, Auckland 1142 (New Zealand); Chang, Chungyu; Ton-That, Hung [University of Texas–Houston Medical School, Houston, TX 77030 (United States); Baker, Edward N., E-mail: ted.baker@auckland.ac.nz [University of Auckland, Private Bag 92019, Auckland 1142 (New Zealand)

    2014-05-01

    Two crystal structures of the major pilin SpaD from C. diphtheriae have been determined at 1.87 and 2.5 Å resolution. The N-terminal domain is found to contain an isopeptide bond that forms slowly over time in the recombinant protein. Given its structural context, this provides insight into the relationship between internal isopeptide-bond formation and pilus assembly. The Gram-positive organism Corynebacterium diphtheriae, the cause of diphtheria in humans, expresses pili on its surface which it uses for adhesion and colonization of its host. These pili are covalent protein polymers composed of three types of pilin subunit that are assembled by specific sortase enzymes. A structural analysis of the major pilin SpaD, which forms the polymeric backbone of one of the three types of pilus expressed by C. diphtheriae, is reported. Mass-spectral and crystallographic analysis shows that SpaD contains three internal Lys–Asn isopeptide bonds. One of these, shown by mass spectrometry to be located in the N-terminal D1 domain of the protein, only forms slowly, implying an energy barrier to bond formation. Two crystal structures, of the full-length three-domain protein at 2.5 Å resolution and of a two-domain (D2-D3) construct at 1.87 Å resolution, show that each of the three Ig-like domains contains a single Lys–Asn isopeptide-bond cross-link, assumed to give mechanical stability as in other such pili. Additional stabilizing features include a disulfide bond in the D3 domain and a calcium-binding loop in D2. The N-terminal D1 domain is more flexible than the others and, by analogy with other major pilins of this type, the slow formation of its isopeptide bond can be attributed to its location adjacent to the lysine used in sortase-mediated polymerization during pilus assembly.

  17. Identification of an additional minor pilin essential for piliation in the archaeon Methanococcus maripaludis.

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    Nair, Divya B; Chung, Daniel K C; Schneider, James; Uchida, Kaoru; Aizawa, Shin-Ichi; Jarrell, Ken F

    2013-01-01

    Methanococcus maripaludis is an archaeon with two studied surface appendages, archaella and type IV-like pili. Previously, the major structural pilin was identified as MMP1685 and three additional proteins were designated as minor pilins (EpdA, EpdB and EpdC). All of the proteins are likely processed by the pilin-specific prepilin peptidase EppA. Six other genes were identified earlier as likely encoding pilin proteins processed also by EppA. In this study, each of the six genes (mmp0528, mmp0600, mmp0601, mmp0709, mmp0903 and mmp1283) was deleted and the mutants examined by electron microscopy to determine their essentiality for pili formation. While mRNA transcripts of all genes were detected by RT-PCR, only the deletion of mmp1283 led to nonpiliated cells. This strain could be complemented back to a piliated state by supplying a wildtype copy of the mmp1283 gene in trans. This study adds to the complexity of the type IV pili system in M. maripaludis and raises questions about the functions of the remaining five pilin-like genes and whether M. maripaludis under other growth conditions may be able to assemble additional pili-like structures.

  18. Identification of an additional minor pilin essential for piliation in the archaeon Methanococcus maripaludis.

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    Divya B Nair

    Full Text Available Methanococcus maripaludis is an archaeon with two studied surface appendages, archaella and type IV-like pili. Previously, the major structural pilin was identified as MMP1685 and three additional proteins were designated as minor pilins (EpdA, EpdB and EpdC. All of the proteins are likely processed by the pilin-specific prepilin peptidase EppA. Six other genes were identified earlier as likely encoding pilin proteins processed also by EppA. In this study, each of the six genes (mmp0528, mmp0600, mmp0601, mmp0709, mmp0903 and mmp1283 was deleted and the mutants examined by electron microscopy to determine their essentiality for pili formation. While mRNA transcripts of all genes were detected by RT-PCR, only the deletion of mmp1283 led to nonpiliated cells. This strain could be complemented back to a piliated state by supplying a wildtype copy of the mmp1283 gene in trans. This study adds to the complexity of the type IV pili system in M. maripaludis and raises questions about the functions of the remaining five pilin-like genes and whether M. maripaludis under other growth conditions may be able to assemble additional pili-like structures.

  19. Structural basis of typhod: Salmonella typhi type IVb pilin (PilS) and cystic fibrosis transmembrane conductance regulator interaction

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    Balakrishna, A.; Saxena, A; Mok, H; Swaminathan, K

    2009-01-01

    The type IVb pilus of the enteropathogenic bacteria Salmonella typhi is a major adhesion factor during the entry of this pathogen into gastrointestinal epithelial cells. Its target of adhesion is a stretch of 10 residues from the first extracellular domain of cystic fibrosis transmembrane conductance regulator (CFTR). The crystal structure of the N-terminal 25 amino acid deleted S. typhi native PilS protein (PilS), which makes the pilus, was determined at 1.9 A resolution by the multiwavelength anomalous dispersion method. Also, the structure of the complex of PilS and a target CFTR peptide, determined at 1.8 A, confirms that residues 113-117 (NKEER) of CFTR are involved in binding with the pilin protein and gives us insight on the amino acids that are essential for binding. Furthermore, we have also explored the role of a conserved disulfide bridge in pilus formation. The subunit structure and assembly architecture are crucial for understanding pilus functions and designing suitable therapeutics against typhoid.

  20. Structural basis of typhoid: Salmonella typhi type IVb pilin (PiLS) and cystic fibrosis transmembrane conductance regulator interaction

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    Balakrishna, A.M.; Saxena, A.; Mok, H. Y.-K.; Swaminathan, K.

    2009-11-01

    The type IVb pilus of the enteropathogenic bacteria Salmonella typhi is a major adhesion factor during the entry of this pathogen into gastrointestinal epithelial cells. Its target of adhesion is a stretch of 10 residues from the first extracellular domain of cystic fibrosis transmembrane conductance regulator (CFTR). The crystal structure of the N-terminal 25 amino acid deleted S. typhi native PilS protein ({Delta}PilS), which makes the pilus, was determined at 1.9 {angstrom} resolution by the multiwavelength anomalous dispersion method. Also, the structure of the complex of {Delta}PilS and a target CFTR peptide, determined at 1.8 {angstrom}, confirms that residues 113-117 (NKEER) of CFTR are involved in binding with the pilin protein and gives us insight on the amino acids that are essential for binding. Furthermore, we have also explored the role of a conserved disulfide bridge in pilus formation. The subunit structure and assembly architecture are crucial for understanding pilus functions and designing suitable therapeutics against typhoid.

  1. Structural Basis of Typhoid: Salmonella typhi Type IVb pilin (PilS) and Cystic Fibrosis Transmembrane Conductance Regulatory Interaction

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    Balakrishna, A.; Saxena, A; Mok, H; Swaminathan, K

    2009-01-01

    The type IVb pilus of the enteropathogenic bacteria Salmonella typhi is a major adhesion factor during the entry of this pathogen into gastrointestinal epithelial cells. Its target of adhesion is a stretch of 10 residues from the first extracellular domain of cystic fibrosis transmembrane conductance regulator (CFTR). The crystal structure of the N-terminal 25 amino acid deleted S. typhi native PilS protein (PilS), which makes the pilus, was determined at 1.9 A resolution by the multiwavelength anomalous dispersion method. Also, the structure of the complex of PilS and a target CFTR peptide, determined at 1.8 A, confirms that residues 113-117 (NKEER) of CFTR are involved in binding with the pilin protein and gives us insight on the amino acids that are essential for binding. Furthermore, we have also explored the role of a conserved disulfide bridge in pilus formation. The subunit structure and assembly architecture are crucial for understanding pilus functions and designing suitable therapeutics against typhoid.

  2. Epithelial Adhesion Mediated by Pilin SpaC Is Required for Lactobacillus rhamnosus GG-Induced Cellular Responses

    Science.gov (United States)

    Ardita, Courtney S.; Mercante, Jeffrey W.; Kwon, Young Man; Luo, Liping; Crawford, Madelyn E.; Powell, Domonica N.; Jones, Rheinallt M.

    2014-01-01

    Lactobacillus rhamnosus GG is a widely used probiotic, and the strain's salutary effects on the intestine have been extensively documented. We previously reported that strain GG can modulate inflammatory signaling, as well as epithelial migration and proliferation, by activating NADPH oxidase 1-catalyzed generation of reactive oxygen species (ROS). However, how strain GG induces these responses is unknown. Here, we report that strain GG's probiotic benefits are dependent on the bacterial-epithelial interaction mediated by the SpaC pilin subunit. By comparing strain GG to an isogenic mutant that lacks SpaC (strain GGΩspaC), we establish that SpaC is necessary for strain GG to adhere to gut mucosa, that SpaC contributes to strain GG-induced epithelial generation of ROS, and that SpaC plays a role in strain GG's capacity to stimulate extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) signaling in enterocytes. In addition, we show that SpaC is required for strain GG-mediated stimulation of cell proliferation and protection against radiologically inflicted intestinal injury. The identification of a critical surface protein required for strain GG to mediate its probiotic influence advances our understanding of the molecular basis for the symbiotic relationship between some commensal bacteria of the gut lumen and enterocytes. Further insights into this relationship are critical for the development of novel approaches to treat intestinal diseases. PMID:24928883

  3. Epithelial adhesion mediated by pilin SpaC is required for Lactobacillus rhamnosus GG-induced cellular responses.

    Science.gov (United States)

    Ardita, Courtney S; Mercante, Jeffrey W; Kwon, Young Man; Luo, Liping; Crawford, Madelyn E; Powell, Domonica N; Jones, Rheinallt M; Neish, Andrew S

    2014-08-01

    Lactobacillus rhamnosus GG is a widely used probiotic, and the strain's salutary effects on the intestine have been extensively documented. We previously reported that strain GG can modulate inflammatory signaling, as well as epithelial migration and proliferation, by activating NADPH oxidase 1-catalyzed generation of reactive oxygen species (ROS). However, how strain GG induces these responses is unknown. Here, we report that strain GG's probiotic benefits are dependent on the bacterial-epithelial interaction mediated by the SpaC pilin subunit. By comparing strain GG to an isogenic mutant that lacks SpaC (strain GGΩspaC), we establish that SpaC is necessary for strain GG to adhere to gut mucosa, that SpaC contributes to strain GG-induced epithelial generation of ROS, and that SpaC plays a role in strain GG's capacity to stimulate extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) signaling in enterocytes. In addition, we show that SpaC is required for strain GG-mediated stimulation of cell proliferation and protection against radiologically inflicted intestinal injury. The identification of a critical surface protein required for strain GG to mediate its probiotic influence advances our understanding of the molecular basis for the symbiotic relationship between some commensal bacteria of the gut lumen and enterocytes. Further insights into this relationship are critical for the development of novel approaches to treat intestinal diseases. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  4. Ancillary Services from Wind Farms

    DEFF Research Database (Denmark)

    Meeting the EU objectives of sustainable energy supply in the near future involves a dramatic increase of the electricity demand covered by variable renewable sources, among which wind power holds an important role. This important role comes together with ever increasing requirements of wind powe...... plants ability of delivering ancillary services to the power system. The presentation attempts at giving an overview of the present (and future) research on the ability of large (offshore) wind farms to provide power system services....

  5. Oligomerized backbone pilin helps piliated Lactococcus lactis to withstand shear flow.

    Science.gov (United States)

    Castelain, Mickaël; Duviau, Marie-Pierre; Oxaran, Virginie; Schmitz, Philippe; Cocaign-Bousquet, Muriel; Loubière, Pascal; Piard, Jean-Christophe; Mercier-Bonin, Muriel

    2016-09-01

    The present work focuses on the role of pili present at the cell surface of Lactococcus lactis in bacterial adhesion to abiotic (hydrophobic polystyrene) and biotic (mucin-coated polystyrene) surfaces. Native pili-displaying strains and isogenic derivatives in which pilins or sortase C structural genes had been modified were used. Surface physico-chemistry, morphology and shear-flow-induced detachment of lactococcal cells were evaluated. The involvement of pili in L. lactis adhesion was clearly demonstrated, irrespective of the surface characteristics (hydrophobic/hydrophilic, presence or not of specific binding sites). The accessory pilin, PilC, and the backbone pilin, PilB, were revealed to play a major role in adhesion, provided that the PilB was present in its polymerized form. Within the population fraction that remained attached to the surface under increasing shear flow, different association behaviors were observed, showing that pili could serve as anchoring sites thus hampering the effect of shear flow on cell orientation and detachment.

  6. Ancillary service provision from distributed generation

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2004-07-01

    This report summarises the findings of a study undertaken to investigate the potential for establishing ancillary service markets at the distribution level in the UK. Existing arrangements for ancillary service markets globally, the design of these markets, regulatory and legislative changes that may be required, different forms of distributed generation (DG), and prospects of increasing the connection to the distributed network are examined along with commercial frameworks and technical procedures, infrastructure requirements, and the effects on different market participants. The scope for new ancillary services at the distribution level, ancillary services from DG, the prospects for DG, commercial and technical aspects, and impact assessments are reviewed.

  7. Ancillary service provision from distributed generation

    International Nuclear Information System (INIS)

    2004-01-01

    This report summarises the findings of a study undertaken to investigate the potential for establishing ancillary service markets at the distribution level in the UK. Existing arrangements for ancillary service markets globally, the design of these markets, regulatory and legislative changes that may be required, different forms of distributed generation (DG), and prospects of increasing the connection to the distributed network are examined along with commercial frameworks and technical procedures, infrastructure requirements, and the effects on different market participants. The scope for new ancillary services at the distribution level, ancillary services from DG, the prospects for DG, commercial and technical aspects, and impact assessments are reviewed

  8. Ancillary Services Provided from DER

    Energy Technology Data Exchange (ETDEWEB)

    Campbell, J.B.

    2005-12-21

    Distributed energy resources (DER) are quickly making their way to industry primarily as backup generation. They are effective at starting and then producing full-load power within a few seconds. The distribution system is aging and transmission system development has not kept up with the growth in load and generation. The nation's transmission system is stressed with heavy power flows over long distances, and many areas are experiencing problems in providing the power quality needed to satisfy customers. Thus, a new market for DER is beginning to emerge. DER can alleviate the burden on the distribution system by providing ancillary services while providing a cost adjustment for the DER owner. This report describes 10 types of ancillary services that distributed generation (DG) can provide to the distribution system. Of these 10 services the feasibility, control strategy, effectiveness, and cost benefits are all analyzed as in the context of a future utility-power market. In this market, services will be provided at a local level that will benefit the customer, the distribution utility, and the transmission company.

  9. Inversion of Moraxella lacunata type 4 pilin gene sequences by a Neisseria gonorrhoeae site-specific recombinase.

    Science.gov (United States)

    Rozsa, F W; Meyer, T F; Fussenegger, M

    1997-01-01

    A plasmid library of Neisseria gonorrhoeae sequences was screened for the ability to mediate recombinations on a sequence containing the Moraxella lacunata type 4 pilin gene invertible region in Escherichia coli. A plasmid containing the N. gonorrhoeae sequence encoding the putative recombinase (gcr) was identified and sequenced. Plasmids containing gcr were able to mediate site-specific recombinations despite a weak amino acid homology to Piv, the native M. lacunata pilin gene invertase. The gcr gene is present only in pathogenic strains of Neisseria tested; however, in our assays gene knockouts of gcr did not alter the variation of surface features that play a role in the pathogenesis of N. gonorrhoeae. PMID:9079926

  10. Purification, crystallization and preliminary crystallographic analysis of the minor pilin FctB from Streptococcus pyogenes

    International Nuclear Information System (INIS)

    Linke, Christian; Young, Paul G.; Kang, Hae Joo; Proft, Thomas; Baker, Edward N.

    2010-01-01

    The minor pilin FctB from S. pyogenes strain 90/306S was expressed in E. coli, purified and crystallized. The hexagonal FctB crystals diffracted to 2.9 Å resolution. The minor pilin FctB is an integral part of the pilus assembly expressed by Streptococcus pyogenes. Since it is located at the cell wall, it can be hypothesized that it functions as a cell-wall anchor for the streptococcal pilus. In order to elucidate its structure, the genes for FctB from the S. pyogenes strains 90/306S and SF370 were cloned for overexpression in Escherichia coli. FctB from strain 90/306S was crystallized by the sitting-drop vapour-diffusion method using sodium citrate as a precipitant. The hexagonal FctB crystals belonged to space group P6 1 or P6 5 , with unit-cell parameters a = b = 95.15, c = 100.25 Å, and diffracted to 2.9 Å resolution

  11. Exogam with its ancillary detectors

    International Nuclear Information System (INIS)

    Theisen, Ch.

    2004-01-01

    The goal of the meeting was to discuss the status of experiments using Exogam and its ancillary detectors. Presentations include details of experiments already run and those in preparation. This document gathers most of the slides presented at the meeting. The results of 7 experiments are described. There are 9 contributions as it follows: 1) presentation of the Vamos spectrometer p.1-15, 2) the Clara (detector) and Prisma (spectrometer) setup: status and perspectives p.16-33, 3) experiment E344aS: shape coexistence near the N=Z line and collective properties of Kr isotopes investigated by low-energy Coulomb excitation p.34-56, 4) experiment E389: study of effective charges at Sm 100 via the nuclear lifetime measurements in Pd 96 and Cd 98 p.57-68, 5) experiment E375: spectroscopy of trans-fermium nuclei p.69-91, 6) experiment E408S: He 8 on Pb 208 and Bi 209 p.92-102, 7) experiment E462: very neutron rich calcium isotopes at N=32, structure of magic Ca 54 studied by gamma-spectroscopy with deep inelastic reactions p.103-115, 8) experiment E451: searching for T=D pairing and a new coupling scheme in Pd 92 and Ru 88 p.116-141, and 9) experiment E404aS: identification of gamma rays in nuclei around the drip-line nucleus Sm 130 , probing the maximally deformed light rare earth region

  12. SMEX03 Ancillary Soil Characteristics Data, Alabama

    Data.gov (United States)

    National Aeronautics and Space Administration — The SMEX03 Ancillary Soil Characteristics data set contains data for the regional study areas of Alabama, Georgia, and Oklahoma, USA as part of the 2003 Soil...

  13. Pressure ulcer risk in ancillary services patients.

    Science.gov (United States)

    Messer, Monica S

    2010-01-01

    Although pressure ulcer prevention has received considerable attention in the literature since the 1960s, little has been published about the risk of pressure injury to patients undergoing procedures in diagnostic and interventional ancillary units such as radiology, renal dialysis, cardiac, and vascular procedure laboratories. In the only research study found in this population, the incidence of pressure injury in patients undergoing lengthy radiology procedures was 53.8%. Therefore, I reviewed the research literature in order to identify which specific factors are most likely to place persons undergoing ancillary procedures at risk for pressure injury. Sufficient evidence was found to implicate high interface pressures on ancillary support surfaces, shear incurred from patient movement and positioning, advanced age, severe neurologic impairment, anesthesia and sedation, fever, sepsis, and hypotension as risk factors for pressure injury while receiving services from interventional ancillary units.

  14. Exogam with its ancillary detectors

    Energy Technology Data Exchange (ETDEWEB)

    Theisen, Ch. [CEA Saclay, Dept. d' Astrophysique, de Physique des Particules de Physique Nucleaire et de l' Instrumentation Associee, 91- Gif sur Yvette (France)

    2004-07-01

    The goal of the meeting was to discuss the status of experiments using Exogam and its ancillary detectors. Presentations include details of experiments already run and those in preparation. This document gathers most of the slides presented at the meeting. The results of 7 experiments are described. There are 9 contributions as it follows: 1) presentation of the Vamos spectrometer p.1-15, 2) the Clara (detector) and Prisma (spectrometer) setup: status and perspectives p.16-33, 3) experiment E344aS: shape coexistence near the N=Z line and collective properties of Kr isotopes investigated by low-energy Coulomb excitation p.34-56, 4) experiment E389: study of effective charges at Sm{sup 100} via the nuclear lifetime measurements in Pd{sup 96} and Cd{sup 98} p.57-68, 5) experiment E375: spectroscopy of trans-fermium nuclei p.69-91, 6) experiment E408S: He{sup 8} on Pb{sup 208} and Bi{sup 209} p.92-102, 7) experiment E462: very neutron rich calcium isotopes at N=32, structure of magic Ca{sup 54} studied by gamma-spectroscopy with deep inelastic reactions p.103-115, 8) experiment E451: searching for T=D pairing and a new coupling scheme in Pd{sup 92} and Ru{sup 88} p.116-141, and 9) experiment E404aS: identification of gamma rays in nuclei around the drip-line nucleus Sm{sup 130}, probing the maximally deformed light rare earth region.

  15. AX Tank Farm ancillary equipment study

    International Nuclear Information System (INIS)

    SKELLY, W.A.

    1999-01-01

    This report examines the feasibility of remediating ancillary equipment associated with the 241-AX Tank Farm at the Hanford Site. Ancillary equipment includes surface structures and equipment, process waste piping, ventilation components, wells, and pits, boxes, sumps, and tanks used to make waste transfers to/from the AX tanks and adjoining tank farms. Two remedial alternatives are considered: (1) excavation and removal of all ancillary equipment items, and (2) in-situ stabilization by grout filling, the 241-AX Tank Farm is being employed as a strawman in engineering studies evaluating clean and landfill closure options for Hanford single-shell tanks. This is one of several reports being prepared for use by the Hanford Tanks Initiative Project to explore potential closure options and to develop retrieval performance evaluation criteria for tank farms

  16. Power system studies of new ancillary services

    DEFF Research Database (Denmark)

    Hansen, Anca Daniela; Altin, Müfit

    The objective of this report is to illustrate and analyse, by means of simulation test cases, the impact of wind power advanced ancillary services, like inertial response (IR), power oscillation damping (POD) and synchronising power (SP) on the power system. Generic models for wind turbine, wind...... power plant and power system are used in the investigation....

  17. Promoting crystallisation of the Salmonella enteritidis fimbriae 14 pilin SefD using deuterium oxide

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Bing; Garnett, James A.; Lee, Wei-chao; Lin, Jing; Salgado, Paula; Taylor, Jonathan; Xu, Yingqi; Lambert, Sebastian; Cota, Ernesto [Centre for Structural Biology and Division of Molecular Biosciences, Imperial College London, South Kensington, London SW7 2AZ (United Kingdom); Matthews, Steve, E-mail: s.j.matthews@imperial.ac.uk [Centre for Structural Biology and Division of Molecular Biosciences, Imperial College London, South Kensington, London SW7 2AZ (United Kingdom)

    2012-05-04

    Highlights: Black-Right-Pointing-Pointer The benefits of D{sub 2}O in screening for crystallisation was explored. Black-Right-Pointing-Pointer The crystal structures of the SefD pilin in both H{sub 2}O and D{sub 2}O reveal differences. Black-Right-Pointing-Pointer Crystallisation improvements are explained by altered interactions in D{sub 2}O crystals. Black-Right-Pointing-Pointer D{sub 2}O is useful additive in sparse-matrix screening for crystallisation. -- Abstract: The use of heavy water (D{sub 2}O) as a solvent is commonplace in many spectroscopic techniques for the study of biological macromolecules. A significant deuterium isotope effect exists where hydrogen-bonding is important, such as in protein stability, dynamics and assembly. Here we illustrate the use of D{sub 2}O in additive screening for the production of reproducible diffraction-quality crystals for the Salmonella enteritidis fimbriae 14 (SEF14) putative tip adhesin, SefD.

  18. Ancillary services provided by PV power plants

    Directory of Open Access Journals (Sweden)

    Antonio PIERNO

    2016-07-01

    Full Text Available Renewable energy sources are widely utilized in distributed generation systems, and, recently, they are also considered for providing ancillary services. The paper is focused on PV plants, a survey of the most interesting papers published in the literature in the last decade is reported and the main characteristics of the technical proposals, with their advantages and limits, are evidenced. The results are schematically shown in a table that immediately gives the opportunity to be aware of what was already done, representing a reference tool.

  19. Truncation of type IV pilin induces mucoidy in Pseudomonas aeruginosa strain PAO579.

    Science.gov (United States)

    Ryan Withers, T; Heath Damron, F; Yin, Yeshi; Yu, Hongwei D

    2013-06-01

    Pseudomonas aeruginosa is a Gram negative, opportunistic pathogen that uses the overproduction of alginate, a surface polysaccharide, to form biofilms in vivo. Overproduction of alginate, also known as mucoidy, affords the bacterium protection from the host's defenses and facilitates the establishment of chronic lung infections in individuals with cystic fibrosis. Expression of the alginate biosynthetic operon is primarily controlled by the alternative sigma factor AlgU (AlgT/σ(22) ). In a nonmucoid strain, AlgU is sequestered by the transmembrane antisigma factor MucA to the cytoplasmic membrane. AlgU can be released from MucA via regulated intramembrane proteolysis by proteases AlgW and MucP causing the conversion to mucoidy. Pseudomonas aeruginosa strain PAO579, a derivative of the nonmucoid strain PAO1, is mucoid due to an unidentified mutation (muc-23). Using whole genome sequencing, we identified 16 nonsynonymous and 15 synonymous single nucleotide polymorphisms (SNP). We then identified three tandem single point mutations in the pilA gene (PA4525), as the cause of mucoidy in PAO579. These tandem mutations generate a premature stop codon resulting in a truncated version of PilA (PilA(108) ), with a C-terminal motif of phenylalanine-threonine-phenylalanine (FTF). Inactivation of pilA(108) confirmed it was required for mucoidy. Additionally, algW and algU were also required for mucoidy of PAO579. Western blot analysis indicated that MucA was less stable in PAO579 than nonmucoid PAO1 or PAO381. The mucoid phenotype and high PalgU and PalgD promoter activities of PAO579 require pilA(108) , algW, algU, and rpoN encoding the alternative sigma factor σ(54) . We also observed that RpoN regulates expression of algW and pilA in PAO579. Together, these results suggest that truncation in type IV pilin in P. aeruginosa strain PAO579 can induce mucoidy through an AlgW/AlgU-dependent pathway. © 2013 The Authors. Microbiology Open published by John Wiley & Sons Ltd.

  20. Industrial Demand Management Providing Ancillary Services to the Distribution Grid

    DEFF Research Database (Denmark)

    Rahnama, Samira; Green, Torben; Lyhne, Casper

    2017-01-01

    A prominent feature of the future smart grid is the active participation of the consumer side in ancillary service provision. Grid operators procure ancillary services, including regulating power, voltage control, frequency control, and so on, to ensure safe, reliable, and high-quality electricity...

  1. 39 CFR 122.1 - Ancillary special services.

    Science.gov (United States)

    2010-07-01

    ... 39 Postal Service 1 2010-07-01 2010-07-01 false Ancillary special services. 122.1 Section 122.1 Postal Service UNITED STATES POSTAL SERVICE POST OFFICE SERVICES [DOMESTIC MAIL] SERVICE STANDARDS FOR MARKET-DOMINANT SPECIAL SERVICES PRODUCTS § 122.1 Ancillary special services. (a) For the market-dominant...

  2. Integration of Flexible Consumers in the Ancillary Service Markets

    DEFF Research Database (Denmark)

    Biegel, Benjamin; Westenholz, Mikkel; Hansen, Lars Henrik

    2014-01-01

    services. In this work we present a simple method based on the existing ancillary service markets that resolves these issues via increased information and communication technology. The method allows an aggregator to continuously utilize the markets for slower ancillary service to ensure that its portfolio...

  3. 16 CFR 1.62 - Ancillary court orders pending review.

    Science.gov (United States)

    2010-01-01

    ... 16 Commercial Practices 1 2010-01-01 2010-01-01 false Ancillary court orders pending review. 1.62 Section 1.62 Commercial Practices FEDERAL TRADE COMMISSION ORGANIZATION, PROCEDURES AND RULES OF PRACTICE GENERAL PROCEDURES Injunctive and Condemnation Proceedings § 1.62 Ancillary court orders pending review...

  4. Illustration of Modern Wind Turbine Ancillary Services

    Directory of Open Access Journals (Sweden)

    Ioannis D. Margaris

    2010-06-01

    Full Text Available Increasing levels of wind power penetration in modern power systems has set intensively high standards with respect to wind turbine technology during the last years. Security issues have become rather critical and operation of wind farms as conventional power plants is becoming a necessity as wind turbines replace conventional units on the production side. This article includes a review of the basic control issues regarding the capability of the Doubly Fed Induction Generator (DFIG wind turbine configuration to fulfill the basic technical requirements set by the system operators and contribute to power system security. An overview of ancillary services provided by wind turbine technology nowadays is provided, i.e., fault ride-through capability, reactive power supply and frequency-active power control.

  5. Structure of Streptococcus agalactiae tip pilin GBS104: a model for GBS pili assembly and host interactions

    Energy Technology Data Exchange (ETDEWEB)

    Krishnan, Vengadesan [UNESCO Regional Centre for Biotechnology (RCB), Gurgaon 122 016, Haryana (India); Dwivedi, Prabhat [University of Texas Health Science Center, Houston, TX 77030 (United States); Kim, Brandon J. [San Diego State University, 5500 Campanile Drive, San Diego, CA 92182 (United States); Samal, Alexandra; Macon, Kevin [University of Alabama at Birmingham, Birmingham, AL 35294 (United States); Ma, Xin; Mishra, Arunima [University of Texas Health Science Center, Houston, TX 77030 (United States); Doran, Kelly S. [San Diego State University, 5500 Campanile Drive, San Diego, CA 92182 (United States); Ton-That, Hung [University of Texas Health Science Center, Houston, TX 77030 (United States); Narayana, Sthanam V. L., E-mail: narayana@uab.edu [University of Alabama at Birmingham, Birmingham, AL 35294 (United States); UNESCO Regional Centre for Biotechnology (RCB), Gurgaon 122 016, Haryana (India)

    2013-06-01

    The crystal structure of a 75 kDa central fragment of GBS104, a tip pilin from the 2063V/R strain of Streptococcus agalactiae (group B streptococcus; GBS), is reported. The crystal structure of a 75 kDa central fragment of GBS104, a tip pilin from the 2063V/R strain of Streptococcus agalactiae (group B streptococcus; GBS), is reported. In addition, a homology model of the remaining two domains of GBS104 was built and a model of full-length GBS104 was generated by combining the homology model (the N1 and N4 domains) and the crystal structure of the 75 kDa fragment (the N2 and N3 domains). This rod-shaped GBS104 model is constructed of three IgG-like domains (the N1, N2 and N4 domains) and one vWFA-like domain (the N3 domain). The N1 and N2 domains of GBS104 are assembled with distinct and remote segments contributed by the N- and C-termini. The metal-binding site in the N3 domain of GBS104 is in the closed/low-affinity conformation. Interestingly, this domain hosts two long arms that project away from the metal-binding site. Using site-directed mutagenesis, two cysteine residues that lock the N3 domain of GBS104 into the open/high-affinity conformation were introduced. Both wild-type and disulfide-locked recombinant proteins were tested for binding to extracellular matrix proteins such as collagen, fibronectin, fibrinogen and laminin, and an increase in fibronectin binding affinity was identified for the disulfide-locked N3 domain, suggesting that induced conformational changes may play a possible role in receptor binding.

  6. Glycosylation of alpha(2)delta(1) subunit: a sweet talk with Ca(v)1.2 channels

    Czech Academy of Sciences Publication Activity Database

    Lazniewska, Joanna; Weiss, Norbert

    2016-01-01

    Roč. 35, č. 3 (2016), s. 239-242 ISSN 0231-5882 R&D Projects: GA ČR GA15-13556S; GA MŠk 7AMB15FR015 Institutional support: RVO:61388963 Keywords : calcium channel * Ca(v)1.2 channel * ancillary subunit * alpha(2)delta(1) subunit * glycosylation * trafficking Subject RIV: CE - Biochemistry Impact factor: 1.170, year: 2016

  7. The N-Terminal GYPSY Motif Is Required for Pilin-Specific Sortase SrtC1 Functionality in Lactobacillus rhamnosus Strain GG.

    Directory of Open Access Journals (Sweden)

    François P Douillard

    Full Text Available Predominantly identified in pathogenic Gram-positive bacteria, sortase-dependent pili are also found in commensal species, such as the probiotic-marketed strain Lactobacillus rhamnosus strain GG. Pili are typically associated with host colonization, immune signalling and biofilm formation. Comparative analysis of the N-terminal domains of pilin-specific sortases from various piliated Gram-positive bacteria identified a conserved motif, called GYPSY, within the signal sequence. We investigated the function and role of the GYPSY residues by directed mutagenesis in homologous (rod-shaped and heterologous (coccoid-shaped expression systems for pilus formation. Substitutions of some of the GYPSY residues, and more specifically the proline residue, were found to have a direct impact on the degree of piliation of Lb. rhamnosus GG. The present findings uncover a new signalling element involved in the functionality of pilin-specific sortases controlling the pilus biogenesis of Lb. rhamnosus GG and related piliated Gram-positive species.

  8. The N-Terminal GYPSY Motif Is Required for Pilin-Specific Sortase SrtC1 Functionality in Lactobacillus rhamnosus Strain GG.

    Science.gov (United States)

    Douillard, François P; Rasinkangas, Pia; Bhattacharjee, Arnab; Palva, Airi; de Vos, Willem M

    2016-01-01

    Predominantly identified in pathogenic Gram-positive bacteria, sortase-dependent pili are also found in commensal species, such as the probiotic-marketed strain Lactobacillus rhamnosus strain GG. Pili are typically associated with host colonization, immune signalling and biofilm formation. Comparative analysis of the N-terminal domains of pilin-specific sortases from various piliated Gram-positive bacteria identified a conserved motif, called GYPSY, within the signal sequence. We investigated the function and role of the GYPSY residues by directed mutagenesis in homologous (rod-shaped) and heterologous (coccoid-shaped) expression systems for pilus formation. Substitutions of some of the GYPSY residues, and more specifically the proline residue, were found to have a direct impact on the degree of piliation of Lb. rhamnosus GG. The present findings uncover a new signalling element involved in the functionality of pilin-specific sortases controlling the pilus biogenesis of Lb. rhamnosus GG and related piliated Gram-positive species.

  9. Redefining Requirements of Ancillary Services for Technology Agnostic Sources

    DEFF Research Database (Denmark)

    Bondy, Daniel Esteban Morales; MacDonald, Jason; Kara, Emre Can

    2018-01-01

    New sources for ancillary services are needed, yet the requirements for service provision in most countries are explicitly formulated for traditional generators. This leads to waste of the potential for new technologies to deliver ancillary services. In order to harness this potential, we propose...... to parameterize the requirements of ancillary services so that reserves can be built by combining the advantageous properties of different technologies. The proposal is exemplified through a laboratory test where it shown that the system needs can be covered through cheaper and smaller reserves....

  10. Backbone dynamics of a bacterially expressed peptide from the receptor binding domain of Pseudomonas aeruginosa pilin strain PAK from heteronuclear 1H-15N NMR spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Campbell, A. Patricia [University of Washington, Department of Medicinal Chemistry, School of Pharmacy (United States); Spyracopoulos, Leo [Department of Biochemistry (Canada); Irvin, Randall T. [University of Alberta, Department of Medical Microbiology and Immunology (Canada); Sykes, Brian D. [Department of Biochemistry (Canada)

    2000-07-15

    The backbone dynamics of a {sup 15}N-labeled recombinant PAK pilin peptide spanning residues 128-144 in the C-terminal receptor binding domain of Pseudomonas aeruginosa pilin protein strain PAK (Lys{sup 128}-Cys-Thr-Ser-Asp-Gln-Asp-Glu-Gln-Phe-Ile-Pro-Lys-Gly-Cys-Ser-Lys{sup 144}) were probed by measurements of {sup 15}N NMR relaxation. This PAK(128-144) sequence is a target for the design of a synthetic peptide vaccine effective against multiple strains of P. aeruginosa infection. The {sup 15}N longitudinal (T{sub 1}) and transverse (T{sub 2}) relaxation rates and the steady-state heteronuclear {l_brace}{sup 1}H{r_brace}-{sup 15}N NOE were measured at three fields (7.04, 11.74 and 14.1 Tesla), five temperatures (5, 10, 15, 20, and 25 deg. C ) and at pH 4.5 and 7.2. Relaxation data was analyzed using both the 'model-free' formalism [Lipari, G. and Szabo, A. (1982) J. Am. Chem. Soc., 104, 4546-4559 and 4559-4570] and the reduced spectral density mapping approach [Farrow, N.A., Szabo, A., Torchia, D.A. and Kay, L.E. (1995) J. Biomol. NMR, 6, 153-162]. The relaxation data, spectral densities and order parameters suggest that the type I and type II {beta}-turns spanning residues Asp{sup 134}-Glu-Gln-Phe{sup 137} and Pro{sup 139}-Lys-Gly-Cys{sup 142}, respectively, are the most ordered and structured regions of the peptide. The biological implications of these results will be discussed in relation to the role that backbone motions play in PAK pilin peptide immunogenicity, and within the framework of developing a pilin peptide vaccine capable of conferring broad immunity across P. aeruginosa strains.

  11. Ancillary Care: From Theory to Practice in International Clinical Research

    Science.gov (United States)

    Pratt, Bridget; Zion, Deborah; Lwin, Khin Maung; Cheah, Phaik Yeong; Nosten, Francois; Loff, Bebe

    2013-01-01

    How international research might contribute to justice in global health has not been substantively addressed by bioethics. This article describes how the provision of ancillary care can link international clinical research to the reduction of global health disparities. It identifies the ancillary care obligations supported by a theory of global justice, showing that Jennifer Ruger’s health capability paradigm requires the delivery of ancillary care to trial participants for a limited subset of conditions that cause severe morbidity and mortality. Empirical research on the Shoklo Malaria Research Unit’s (SMRU) vivax malaria treatment trial was then undertaken to demonstrate whether and how these obligations might be upheld in a resource-poor setting. Our findings show that fulfilment of the ancillary care obligations is feasible where there is commitment from chief investigators and funders and is strongly facilitated by SMRU’s dual role as a research unit and medical non-governmental organization. PMID:23864908

  12. Options for pricing ancillary services in a deregulated power system

    Science.gov (United States)

    Yamin, Hatim Yahya

    2001-07-01

    GENCOs in restructured systems are compensated for selling energy in the market. In a restructured market, a mechanism is required to entice participants in the market to provide ancillary services and to ensure adequate compensation that would guarantee its economic viability. The ISO controls the dispatch of generation, manages the reliability of the transmission grid, provides open access to the transmission, buys and provides ancillary services as required, coordinates day-ahead, hour-ahead schedules and performs real time balancing of load and generation, settles real time imbalances and ancillary services sales and purchases. The ISO, also, administers congestion management protocols for the transmission grid. Since the ISO does not own any generating units it must ensure that there is enough reserves for maintaining reliability according to FERC regulations, and sufficient unloaded generating capacity for balancing services in a real-time market. The ISO could meet these requirements by creating a competitive market for ancillary services, which are metered and remain unbundled to provide an accurate compensation for each supplier and cost to each consumer, In this study, we give an overview for restructuring and ancillary services in a restructured power marketplace. Also, we discuss the effect of GENCOs' actions in the competitive energy and ancillary service markets. In addition, we propose an auction market design for hedging ancillary service costs in California market. Furthermore, we show how to include the n-1 and voltage contingencies in security constrained unit commitment. Finally, we present two approaches for GENCOs' unit commitment in a restructured power market; one is based on game theory and the other is based on market price forecasting. In each of the two GENCOs' unit commitment approaches, we discuss the GENCOs' optimal bidding strategies in energy and ancillary service markets to maximize the GENCOs' profit.

  13. Survey of U.S. Ancillary Services Markets

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Zhi [Argonne National Lab. (ANL), Argonne, IL (United States); Levin, Todd [Argonne National Lab. (ANL), Argonne, IL (United States); Conzelmann, Guenter [Argonne National Lab. (ANL), Argonne, IL (United States)

    2016-01-01

    In addition to providing energy to end-consumers, power system operators are also responsible for ensuring system reliability. To this end, power markets maintain an array of ancillary services to ensure it is always possible to balance the supply and demand for energy in real-time. A subset of these ancillary services are commonly procured through market-based mechanisms: namely, Regulation, Spinning, and Non-spinning Reserves.

  14. Off-pathway assembly of fimbria subunits is prevented by chaperone CfaA of CFA/I fimbriae from enterotoxigenic E. coli.

    Science.gov (United States)

    Bao, Rui; Liu, Yang; Savarino, Stephen J; Xia, Di

    2016-12-01

    The assembly of the class 5 colonization factor antigen I (CFA/I) fimbriae of enterotoxigenic E. coli was proposed to proceed via the alternate chaperone-usher pathway. Here, we show that in the absence of the chaperone CfaA, CfaB, the major pilin subunit of CFA/I fimbriae, is able to spontaneously refold and polymerize into cyclic trimers. CfaA kinetically traps CfaB to form a metastable complex that can be stabilized by mutations. Crystal structure of the stabilized complex reveals distinctive interactions provided by CfaA to trap CfaB in an assembly competent state through donor-strand complementation (DSC) and cleft-mediated anchorage. Mutagenesis indicated that DSC controls the stability of the chaperone-subunit complex and the cleft-mediated anchorage of the subunit C-terminus additionally assist in subunit refolding. Surprisingly, over-stabilization of the chaperone-subunit complex led to delayed fimbria assembly, whereas destabilizing the complex resulted in no fimbriation. Thus, CfaA acts predominantly as a kinetic trap by stabilizing subunit to avoid its off-pathway self-polymerization that results in energetically favorable trimers and could serve as a driving force for CFA/I pilus assembly, representing an energetic landscape unique to class 5 fimbria assembly. Published 2016. This article is a U.S. Government work and is in the public domain in the USA. Molecular Microbiology published by John Wiley & Sons Ltd.

  15. Signal peptide prediction suggests Mycobacterium tuberculosis curli pilin subunit secretion via the Sec pathway may hinder MTP overexpression in Escherichia coli

    International Nuclear Information System (INIS)

    Naidoo, N.; Pillay, B.; Bubb, M.; Kumar, A.; Chiliza, T.; Pillay, M.

    2017-01-01

    Introduction Mycobacterium tuberculosis curli pili (MTP) are novel, potential TB diagnostic biomarkers as they are important virulence attributes, unique to the M. tuberculosis complex (MTBC). The production of high quality recombinant transmembrane and secretory proteins that can serve as biomarkers may be challenging due to their secretion attributes. For example, the signal peptide of MTP governed by the classical secretion pathway may hinder the purification of the protein in E. coli systems. In this study, the secretion characteristics of MTP were determined and the cloning, expression and purification of MTP was attempted in E.coli. Materials and methods A fragment of MTP unique to MTBC was cloned into pet101 and pGEX-6P-1 vectors. The clones were confirmed by nucleotide sequencing and expression of the protein was attempted at IPTG concentrations ranging from 0.1mM to 1mM and at temperatures between 25 °C to 37 °C. The pGEX-6P-1/mtp clone expressed protein was purified, yielding a MTP-GST fusion protein and a free GST band that were analysed by LC/MS mass spectrometry. Inclusion body preparation attempted from the pet101/mtp clone yielded two bands at 10 kDa and below 10 kDa, both of which were analysed by LC/MS mass spectrometry. Transcription activity of both the clones was interrogated by real time PCR on the cDNA derived from the induced clones at various time points after induction with IPTG. The signal peptide and protein secretion characteristics of the MTP protein were determined by bioinformatics analysis of the amino acid sequence using publically available software. Results The truncated MTP fragments were successfully cloned in both the vectors as confirmed by nucleotide sequencing. Expression of the pGEX-6P-1/mtp clone using 0.5 mM IPTG at 27 °C demonstrated the presence of the expected fragment at approximately 35 kDa. This was confirmed by Western Blotting using anti-GST antibodies. However, purification of MTP in adequate quantities as a pure protein fraction was unsuccessful. Mass spectrometry did not detect any fragments belonging to MTP, but only those of E.coli membrane proteins for the pet101/mtp clone and fragments of the GST tag in the case of the pGEX-6P-1/mtp clone. The bioinformatics secretion analyses of MTP predicted a strong Sec regulated secretion pathway and the absence of non-classical “mycobacterial specific” secretion. Discussion M. tuberculosis membrane and secretory proteins often contain signal peptides. In this study, excluding the signal peptide region and using a GST tag greatly enhanced the expression of the protein in the soluble fraction. However, purification of the MTP peptide remained problematic due to a lower available peptide concentration resulting from the lower molecular weight, in the purified fraction compared to the GST tag. Alternately, the predicted Sec regulated secretion pathway may play a role in inhibition of MTP overexpression in E.coli. Thus, alternatives to E. coli expression systems or more efficient purification strategies are required for the acquisition of high quality M. tuberculosis antigens.

  16. Signal peptide prediction suggests Mycobacterium tuberculosis curli pilin subunit secretion via the Sec pathway may hinder MTP overexpression in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Natasha Naidoo

    2017-07-01

    Full Text Available Introduction Mycobacterium tuberculosis curli pili (MTP are novel, potential TB diagnostic biomarkers as they are important virulence attributes, unique to the M. tuberculosis complex (MTBC. The production of high quality recombinant transmembrane and secretory proteins that can serve as biomarkers may be challenging due to their secretion attributes. For example, the signal peptide of MTP governed by the classical secretion pathway may hinder the purification of the protein in E. coli systems. In this study, the secretion characteristics of MTP were determined and the cloning, expression and purification of MTP was attempted in E.coli.  Materials and methods A fragment of MTP unique to MTBC was cloned into pet101 and pGEX-6P-1 vectors. The clones were confirmed by nucleotide sequencing and expression of the protein was attempted at IPTG concentrations ranging from 0.1mM to 1mM and at temperatures between 25 °C to 37 °C. The pGEX-6P-1/mtp clone expressed protein was purified, yielding a MTP-GST fusion protein and a free GST band that were analysed by LC/MS mass spectrometry. Inclusion body preparation attempted from the pet101/mtp clone yielded two bands at 10 kDa and below 10 kDa, both of which were analysed by LC/MS mass spectrometry. Transcription activity of both the clones was interrogated by real time PCR on the cDNA derived from the induced clones at various time points after induction with IPTG. The signal peptide and protein secretion characteristics of the MTP protein were determined by bioinformatics analysis of the amino acid sequence using publically available software.  Results The truncated MTP fragments were successfully cloned in both the vectors as confirmed by nucleotide sequencing. Expression of the pGEX-6P-1/mtp clone using 0.5 mM IPTG at 27 °C demonstrated the presence of the expected fragment at approximately 35 kDa. This was confirmed by Western Blotting using anti-GST antibodies. However, purification of MTP in adequate quantities as a pure protein fraction was unsuccessful. Mass spectrometry did not detect any fragments belonging to MTP, but only those of E.coli membrane proteins for the pet101/mtp clone and fragments of the GST tag in the case of the pGEX-6P-1/mtp clone. The bioinformatics secretion analyses of MTP predicted a strong Sec regulated secretion pathway and the absence of non-classical “mycobacterial specific” secretion.  Discussion M. tuberculosis membrane and secretory proteins often contain signal peptides. In this study, excluding the signal peptide region and using a GST tag greatly enhanced the expression of the protein in the soluble fraction. However, purification of the MTP peptide remained problematic due to a lower available peptide concentration resulting from the lower molecular weight, in the purified fraction compared to the GST tag. Alternately, the predicted Sec regulated secretion pathway may play a role in inhibition of MTP overexpression in E.coli. Thus, alternatives to E. coli expression systems or more efficient purification strategies are required for the acquisition of high quality M. tuberculosis antigens.

  17. Signal peptide prediction suggests Mycobacterium tuberculosis curli pilin subunit secretion via the Sec pathway may hinder MTP overexpression in Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Naidoo, N.; Pillay, B.; Bubb, M.; Kumar, A.; Chiliza, T.; Pillay, M.

    2017-07-01

    Introduction Mycobacterium tuberculosis curli pili (MTP) are novel, potential TB diagnostic biomarkers as they are important virulence attributes, unique to the M. tuberculosis complex (MTBC). The production of high quality recombinant transmembrane and secretory proteins that can serve as biomarkers may be challenging due to their secretion attributes. For example, the signal peptide of MTP governed by the classical secretion pathway may hinder the purification of the protein in E. coli systems. In this study, the secretion characteristics of MTP were determined and the cloning, expression and purification of MTP was attempted in E.coli. Materials and methods A fragment of MTP unique to MTBC was cloned into pet101 and pGEX-6P-1 vectors. The clones were confirmed by nucleotide sequencing and expression of the protein was attempted at IPTG concentrations ranging from 0.1mM to 1mM and at temperatures between 25 °C to 37 °C. The pGEX-6P-1/mtp clone expressed protein was purified, yielding a MTP-GST fusion protein and a free GST band that were analysed by LC/MS mass spectrometry. Inclusion body preparation attempted from the pet101/mtp clone yielded two bands at 10 kDa and below 10 kDa, both of which were analysed by LC/MS mass spectrometry. Transcription activity of both the clones was interrogated by real time PCR on the cDNA derived from the induced clones at various time points after induction with IPTG. The signal peptide and protein secretion characteristics of the MTP protein were determined by bioinformatics analysis of the amino acid sequence using publically available software. Results The truncated MTP fragments were successfully cloned in both the vectors as confirmed by nucleotide sequencing. Expression of the pGEX-6P-1/mtp clone using 0.5 mM IPTG at 27 °C demonstrated the presence of the expected fragment at approximately 35 kDa. This was confirmed by Western Blotting using anti-GST antibodies. However, purification of MTP in adequate quantities as a pure protein fraction was unsuccessful. Mass spectrometry did not detect any fragments belonging to MTP, but only those of E.coli membrane proteins for the pet101/mtp clone and fragments of the GST tag in the case of the pGEX-6P-1/mtp clone. The bioinformatics secretion analyses of MTP predicted a strong Sec regulated secretion pathway and the absence of non-classical “mycobacterial specific” secretion. Discussion M. tuberculosis membrane and secretory proteins often contain signal peptides. In this study, excluding the signal peptide region and using a GST tag greatly enhanced the expression of the protein in the soluble fraction. However, purification of the MTP peptide remained problematic due to a lower available peptide concentration resulting from the lower molecular weight, in the purified fraction compared to the GST tag. Alternately, the predicted Sec regulated secretion pathway may play a role in inhibition of MTP overexpression in E.coli. Thus, alternatives to E. coli expression systems or more efficient purification strategies are required for the acquisition of high quality M. tuberculosis antigens.

  18. Ancillary effects of selected acid deposition control policies

    Energy Technology Data Exchange (ETDEWEB)

    Moe, R.J.; Lyke, A.J.; Nesse, R.J.

    1986-08-01

    NAPAP is examining a number of potential ways to reduce the precursors (sulfur dioxide and nitrogen oxides) to acid deposition. However, the policies to reduce acid deposition will have other physical, biological and economic effects unrelated to acid deposition. For example, control policies that reduce sulfur dioxide emissions may also increase visibility. The effects of an acid deposition policy that are unrelated to acid deposition are referred to as ''ancillary'' effects. This reserch identifies and characterizes the principle physical and economic ancillary effects associated with acid deposition control and mitigation policies. In this study the ancillary benefits associated with four specific acid deposition policy options were investigated. The four policy options investigated are: (1) flue gas desulfurization, (2) coal blending or switching, (3) reductions in automobile emissions of NO/sub x/, and (4) lake liming. Potential ancillary benefits of each option were identified and characterized. Particular attention was paid to the literature on economic valuation of potential ancillary effects.

  19. Improving recombinant Rubisco biogenesis, plant photosynthesis and growth by coexpressing its ancillary RAF1 chaperone.

    Science.gov (United States)

    Whitney, Spencer M; Birch, Rosemary; Kelso, Celine; Beck, Jennifer L; Kapralov, Maxim V

    2015-03-17

    Enabling improvements to crop yield and resource use by enhancing the catalysis of the photosynthetic CO2-fixing enzyme Rubisco has been a longstanding challenge. Efforts toward realization of this goal have been greatly assisted by advances in understanding the complexities of Rubisco's biogenesis in plastids and the development of tailored chloroplast transformation tools. Here we generate transplastomic tobacco genotypes expressing Arabidopsis Rubisco large subunits (AtL), both on their own (producing tob(AtL) plants) and with a cognate Rubisco accumulation factor 1 (AtRAF1) chaperone (producing tob(AtL-R1) plants) that has undergone parallel functional coevolution with AtL. We show AtRAF1 assembles as a dimer and is produced in tob(AtL-R1) and Arabidopsis leaves at 10-15 nmol AtRAF1 monomers per square meter. Consistent with a postchaperonin large (L)-subunit assembly role, the AtRAF1 facilitated two to threefold improvements in the amount and biogenesis rate of hybrid L8(A)S8(t) Rubisco [comprising AtL and tobacco small (S) subunits] in tob(AtL-R1) leaves compared with tob(AtL), despite >threefold lower steady-state Rubisco mRNA levels in tob(AtL-R1). Accompanying twofold increases in photosynthetic CO2-assimilation rate and plant growth were measured for tob(AtL-R1) lines. These findings highlight the importance of ancillary protein complementarity during Rubisco biogenesis in plastids, the possible constraints this has imposed on Rubisco adaptive evolution, and the likely need for such interaction specificity to be considered when optimizing recombinant Rubisco bioengineering in plants.

  20. Loads Providing Ancillary Services: Review of International Experience

    Energy Technology Data Exchange (ETDEWEB)

    Heffner, Grayson; Goldman, Charles; Kintner-Meyer, Michael

    2007-05-01

    In this study, we examine the arrangements for and experiences of end-use loads providing ancillary services (AS) in five electricity markets: Australia, the United Kingdom (UK), the Nordic market, and the ERCOT and PJM markets in the United States. Our objective in undertaking this review of international experience was to identify specific approaches or market designs that have enabled customer loads to effectively deliver various ancillary services (AS) products. We hope that this report will contribute to the ongoing discussion in the U.S. and elsewhere regarding what institutional and technical developments are needed to ensure that customer loads can meaningfully participate in all wholesale electricity markets.

  1. EV and HP Providing Ancillary Services in the Nordic Region

    DEFF Research Database (Denmark)

    Liu, Zhaoxi; Wu, Qiuwei

    This report covers the analysis of the electric vehicle (EV) and heat pump (HP) providing ancillary services to the power system of the Nordic region including Denmark, Finland, Norway and Sweden. The analysis is to investigate the feasibility of EVs and HPs to serve as demand as frequency reserve...

  2. Impact of advanced wind power ancillary services on power system

    DEFF Research Database (Denmark)

    Hansen, Anca Daniela; Altin, Müfit

    The objective of this report is to illustrate and analyse, by means of simulation test cases, the impact of wind power advanced ancillary services, like inertial response (IR), power oscillation damping (POD) and synchronising power (SP) on the power system. Generic models for wind turbine, wind...... power plant and power system are used in the investigation....

  3. Long-term pavement performance ancillary information management system (AIMS) reference guide.

    Science.gov (United States)

    2012-11-01

    This document provides information on the Long-Term Pavement Performance (LTPP) program ancillary information. : Ancillary information includes data, images, reference materials, resource documents, and other information that : support and extend the...

  4. Review of Standby and Ancillary Services in the Context of Behind-the-Meter Photovoltaics

    Energy Technology Data Exchange (ETDEWEB)

    Gagnon, Pieter J [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Holm, Alison [National Renewable Energy Laboratory (NREL), Golden, CO (United States)

    2018-04-25

    The New Mexico Public Regulatory Commission (NMPRC) requested information on the following topics: Overview of ancillary and standby services. Examples of how ancillary and standby services are defined, used, and analyzed in other jurisdictions. Considerations regarding how ancillary and standby services may be economically valued. Background information on how ancillary and standby services differ from fixed costs to serve owners of behind-the-meter solar photovoltaic (BTM PV) systems.

  5. Exogam with Vamos, Tiara and other ancillary detectors

    International Nuclear Information System (INIS)

    Lewitowicz, M.; Olivier, L.; Savajols, H.; Catford, W.; Raine, B.; Oliveira, F. de; Perru, O.; Podolyak, Z.; Redon, N.; Bouchez, E.; Simenel, C.; Stanoiu, M.; Lemmon, R.; Rubio, B.; Obertelli, A.; Cederwall, B.; Chapman, R.; Korten, W.; Rubio Barroso, B.

    2003-01-01

    The aim of this workshop was to discuss the status of experiments at GANIL facility using Exogam with Vamos, Tiara and other ancillary detectors. This document gathers most of the slides that were presented at the workshop, it concerns 8 presentations: 1) the status of Spiral and Spiral-II, 2) the status of Tiara (Tiara array is used for the study of nuclear transfer reactions with radioactive beams in inverse kinematics), 3) data acquisition for the ancillary detectors, 4) the identification of gamma-ray in nuclei around Sm 130 : probing the maximally deformed light rare earth region, 5) the Coulomb excitation of radioactive krypton beams (Kr 76 , Kr 74 ), 6) the influence of weakly bound neutrons on fusion around the Coulomb barrier, 7) single particle transfer on doubly-magic Ni 56 , and 8) the study of N=16 for neutron rich nuclei with Ne 26 (d,p)Ne 27 transfer reaction

  6. Advanced Pumped Storage Hydropower and Ancillary Services Provision

    Energy Technology Data Exchange (ETDEWEB)

    Muljadi, Eduard; Gevorgian, Vahan; Mohanpurkar, Manish; Luo, Yusheng; Hovsapian, Rob; Koritarov, Vladimir

    2017-07-12

    This paper presents a high-level overview of the capability of advanced pumped storage hydropower to provide ancillary services including frequency regulation and oscillation damping. Type 3 and Type 4 generators are discussed. The examples given are for a small power system that uses a diesel generator as the main generator and a very large system that uses a gas turbine as the main generator.

  7. Screening and Flagging of Solar Irradiation and Ancillary Meteorological Data

    OpenAIRE

    Geuder, Norbert; Wolfertstetter, Fabian; Wilbert, Stefan; Schüler, David; Affolter, Roman; Kraas, Birk; Lüpfert, Eckhard; Espinar, Bella

    2014-01-01

    International audience; Solar irradiance and ancillary meteorological data is frequently measured by automatic weather stations for use within solar resource assessment for solar power plants. High accuracy measurement data are required for comparison and adjustment of satellite data and derivation of the expectable long-term mean value of the solar resource. Thus, utmost diligence must be taken during the measurement process and data evaluation to achieve data quality required for project fi...

  8. 77 FR 25105 - Reporting of Ancillary Airline Passenger Revenues

    Science.gov (United States)

    2012-04-27

    ...This document announces a public meeting on a Notice of Proposed Rulemaking (NPRM) issued on July 15, 2011. The NPRM proposed changes regarding reporting of airline ancillary passenger revenues, computation of mishandled baggage rates, and collection of separate statistics for mishandled wheelchairs and scooters used by passengers with disabilities. During the public meeting, DOT staff will provide a summary of the proposals in the NPRM and seek input on costs and benefits associated with the implementation of the proposals.

  9. Optimal generator bidding strategies for power and ancillary services

    Science.gov (United States)

    Morinec, Allen G.

    As the electric power industry transitions to a deregulated market, power transactions are made upon price rather than cost. Generator companies are interested in maximizing their profits rather than overall system efficiency. A method to equitably compensate generation providers for real power, and ancillary services such as reactive power and spinning reserve, will ensure a competitive market with an adequate number of suppliers. Optimizing the generation product mix during bidding is necessary to maximize a generator company's profits. The objective of this research work is to determine and formulate appropriate optimal bidding strategies for a generation company in both the energy and ancillary services markets. These strategies should incorporate the capability curves of their generators as constraints to define the optimal product mix and price offered in the day-ahead and real time spot markets. In order to achieve such a goal, a two-player model was composed to simulate market auctions for power generation. A dynamic game methodology was developed to identify Nash Equilibria and Mixed-Strategy Nash Equilibria solutions as optimal generation bidding strategies for two-player non-cooperative variable-sum matrix games with incomplete information. These games integrated the generation product mix of real power, reactive power, and spinning reserve with the generators's capability curves as constraints. The research includes simulations of market auctions, where strategies were tested for generators with different unit constraints, costs, types of competitors, strategies, and demand levels. Studies on the capability of large hydrogen cooled synchronous generators were utilized to derive useful equations that define the exact shape of the capability curve from the intersections of the arcs defined by the centers and radial vectors of the rotor, stator, and steady-state stability limits. The available reactive reserve and spinning reserve were calculated given a

  10. Cyclic Di-GMP Binding by an Assembly ATPase (PilB2) and Control of Type IV Pilin Polymerization in the Gram-Positive Pathogen Clostridium perfringens.

    Science.gov (United States)

    Hendrick, William A; Orr, Mona W; Murray, Samantha R; Lee, Vincent T; Melville, Stephen B

    2017-05-15

    The Gram-positive pathogen Clostridium perfringens possesses type IV pili (TFP), which are extracellular fibers that are polymerized from a pool of pilin monomers in the cytoplasmic membrane. Two proteins that are essential for pilus functions are an assembly ATPase (PilB) and an inner membrane core protein (PilC). Two homologues each of PilB and PilC are present in C. perfringens , called PilB1/PilB2 and PilC1/PilC2, respectively, along with four pilin proteins, PilA1 to PilA4. The gene encoding PilA2, which is considered the major pilin based on previous studies, is immediately downstream of the pilB2 and pilC2 genes. Purified PilB2 had ATPase activity, bound zinc, formed hexamers even in the absence of ATP, and bound the second messenger molecule cyclic di-GMP (c-di-GMP). Circular dichroism spectroscopy of purified PilC2 indicated that it retained its predicted degree of alpha-helical secondary structure. Even though no direct interactions between PilB2 and PilC2 could be detected in vivo or in vitro even in the presence of c-di-GMP, high levels of expression of a diguanylate cyclase from C. perfringens (CPE1788) stimulated polymerization of PilA2 in a PilB2- and PilC2-dependent manner. These results suggest that PilB2 activity is controlled by c-di-GMP levels in vivo but that PilB2-PilC2 interactions are either transitory or of low affinity, in contrast to results reported previously from in vivo studies of the PilB1/PilC1 pair in which PilC1 was needed for polar localization of PilB1. This is the first biochemical characterization of a c-di-GMP-dependent assembly ATPase from a Gram-positive bacterium. IMPORTANCE Type IV pili (TFP) are protein fibers involved in important bacterial functions, including motility, adherence to surfaces and host cells, and natural transformation. All clostridia whose genomes have been sequenced show evidence of the presence of TFP. The genetically tractable species Clostridium perfringens was used to study proteins involved in

  11. Simulation of an arbitrary quantum channel with minimal ancillary resource

    Science.gov (United States)

    Shen, Chao; Noh, Kyungjoo; Albert, Victor V.; Devoret, Michel H.; Schoelkopf, Robert J.; Girvin, Steven M.; Jiang, Liang

    2016-05-01

    We discuss an explicit and efficient construction of quantum circuits that can simulate an arbitrary given quantum channel acting on a d-level quantum system, with the minimal quantum ancillary resource--a qubit and its QND readout. The elementary operations required are unitary evolutions and single qubit projective measurement. We further show that this technique opens up exciting new possibilities in the field of quantum control, quantum simulation, quantum error correction, and quantum state discrimination. Our proposal can be implemented on platforms such as a superconducting transmon qubit inside a microwave cavity.

  12. Transactive Control and Coordination of Distributed Assets for Ancillary Services

    Energy Technology Data Exchange (ETDEWEB)

    Subbarao, Krishnappa [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Fuller, Jason C. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Kalsi, Karanjit [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Somani, Abhishek [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Pratt, Robert G. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Widergren, Steven E. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Chassin, David P. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2013-09-18

    The need to diversify energy supplies, the need to mitigate energy-related environmental impact, and the entry of electric vehicles in large numbers present challenges and opportunities to power system professionals. Wind and solar power provide many benefits, and to reap the benefits the resulting increased variability—forecasted as well as unforecasted—should be addressed. A majority of states and the District of Columbia, representing over half of the total load, have passed renewable portfolio standards. California’s plans call for 33% renewable energy by 2020. For grid balancing and for meeting reliability standards, ancillary services are needed. The needs for these services are poised to increase significantly. Demand resources are receiving increasing attention as one means of providing the ancillary services. Control and coordination of a large number (~millions) of distributed smart grid assets requires innovative approaches. One such approach is transactive control and coordination (TC2)—a distributed, hierarchical, agent-based incentive and control system. The TC2 paradigm is to create a market-like control system in which participation is voluntary and the participant sets the price for participation. For transactions that are frequent, automation of bids and responses is necessary. Such an approach has been developed and demonstrated at the Pacific Northwest National Laboratory. The devices, typically thermostatically controlled heating, ventilating, and air conditioning (HVAC) loads, send their bids—the quantity of energy they need and, based on the consumer preferences encoded in a simple user interface, the price they are willing to pay. The typical bid period is 5 minutes. By aggregating all the bids, a demand curve is generated by the aggregating entity, and matched with a supply curve or supply constraint. The aggregator transmits the clearing price to the devices. The winning devices proceed to consume the energy they bid for and won

  13. Dimerization of the type IV pilin from Pseudomonas aeruginosa strain K122-4 results in increased helix stability as measured by time-resolved hydrogen-deuterium exchange

    Directory of Open Access Journals (Sweden)

    Cristina Lento

    2016-01-01

    Full Text Available Truncated pilin monomers from Pseudomonas aeruginosa strain K122-4 (ΔK122 have been shown to enter a monomer-dimer equilibrium in solution prior to oligomerization into protein nanotubes. Here, we examine the structural changes occurring between the monomeric and dimeric states of ΔK122 using time-resolved hydrogen-deuterium exchange mass spectrometry. Based on levels of deuterium uptake, the N-terminal α-helix and the loop connecting the second and third strands of the anti-parallel β-sheet contribute significantly to pilin dimerization. Conversely, the antiparallel β-sheet and αβ loop region exhibit increased flexibility, while the receptor binding domain retains a rigid conformation in the equilibrium state.

  14. LANDMET Ancillary Soil Moisture data L3 V1 (LANDMET_ANC_SM) at GES DISC

    Data.gov (United States)

    National Aeronautics and Space Administration — This ancillary climatology soil porosity and wetlands coverage information were derived from the daily GEWEX fusion of satellite active and passive microwave...

  15. Ancillary services and optimal household energy management with photovoltaic production

    International Nuclear Information System (INIS)

    Clastres, C.; Ha Pham, T.T.; Wurtz, F.; Bacha, S.

    2010-01-01

    This article presents a project designed to increase the monetary value of photovoltaic (PV) solar production for residential applications. To contribute to developing new functionalities for this type of PV system and an efficient control system for optimising its operation, this article explains how the proposed system could contract to provide ancillary services, particularly the supply of active power services. This provision of service by a PV-based system for domestic applications, not currently available, has prompted a market design proposal related to the distribution system. The mathematical model for calculating the system's optimal operation (sources, load and exchanges of power with the grid) results in a linear mix integer optimisation problem in which the objective is to maximise the profits achieved by taking part in the electricity market. Our approach is illustrated in a case study. PV producers could gain by taking part in the markets for balancing power or ancillary services despite the negative impact on profit of several types of uncertainty, notably the intermittent nature of the PV source.

  16. Exogam with Vamos, Tiara and other ancillary detectors

    Energy Technology Data Exchange (ETDEWEB)

    Lewitowicz, M.; Olivier, L.; Savajols, H.; Catford, W.; Raine, B.; Oliveira, F. de; Perru, O.; Podolyak, Z.; Redon, N.; Bouchez, E.; Simenel, C.; Stanoiu, M.; Lemmon, R.; Rubio, B.; Obertelli, A.; Cederwall, B.; Chapman, R.; Korten, W.; Rubio Barroso, B

    2003-07-01

    The aim of this workshop was to discuss the status of experiments at GANIL facility using Exogam with Vamos, Tiara and other ancillary detectors. This document gathers most of the slides that were presented at the workshop, it concerns 8 presentations: 1) the status of Spiral and Spiral-II, 2) the status of Tiara (Tiara array is used for the study of nuclear transfer reactions with radioactive beams in inverse kinematics), 3) data acquisition for the ancillary detectors, 4) the identification of gamma-ray in nuclei around Sm{sup 130}: probing the maximally deformed light rare earth region, 5) the Coulomb excitation of radioactive krypton beams (Kr{sup 76}, Kr{sup 74}), 6) the influence of weakly bound neutrons on fusion around the Coulomb barrier, 7) single particle transfer on doubly-magic Ni{sup 56}, and 8) the study of N=16 for neutron rich nuclei with Ne{sup 26}(d,p)Ne{sup 27} transfer reaction.

  17. Public attitudes toward ancillary information revealed by pharmacogenetic testing under limited information conditions.

    Science.gov (United States)

    Haga, Susanne B; O'Daniel, Julianne M; Tindall, Genevieve M; Lipkus, Isaac R; Agans, Robert

    2011-08-01

    Pharmacogenetic testing can inform drug dosing and selection by aiding in estimating a patient's genetic risk of adverse response and/or failure to respond. Some pharmacogenetic tests may generate ancillary clinical information unrelated to the drug treatment question for which testing is done-an informational "side effect." We aimed to assess public interest and concerns about pharmacogenetic tests and ancillary information. We conducted a random-digit-dial phone survey of a sample of the US public. We achieved an overall response rate of 42% (n = 1139). When the potential for ancillary information was presented, 85% (±2.82%) of respondents expressed interest in pharmacogenetic testing, compared with 82% (±3.02%) before discussion of ancillary information. Most respondents (89% ± 2.27%) indicated that physicians should inform patients that a pharmacogenetic test may reveal ancillary risk information before testing is ordered. Respondents' interest in actually learning of the ancillary risk finding significantly differed based on disease severity, availability of an intervention, and test validity, even after adjusting for age, gender, education, and race. Under the limited information conditions presented in the survey, the potential of ancillary information does not negatively impact public interest in pharmacogenetic testing. Interest in learning ancillary information is well aligned with the public's desire to be informed about potential benefits and risks before testing, promoting patient autonomy.

  18. 40 CFR 267.197 - What are the requirements for ancillary equipment?

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 26 2010-07-01 2010-07-01 false What are the requirements for ancillary equipment? 267.197 Section 267.197 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY... OPERATING UNDER A STANDARDIZED PERMIT Tank Systems § 267.197 What are the requirements for ancillary...

  19. Rejuvenation of the neck with liposuction and ancillary techniques.

    Science.gov (United States)

    Stebbins, William G; Hanke, C William

    2011-01-01

    An aesthetically pleasing neck is an important component of physical appearance and a frequently targeted area for a variety of rejuvenative procedures. In appropriately selected patients, liposuction of the neck using tumescent local anesthesia can effectively render a more youthful appearance to the anterior and lateral neck by removing superficial adipose tissue and redraping the skin. This article will review all aspects of neck liposuction, including neck and lower face anatomy, proper patient selection and evaluation, necessary equipment, as well as all peri-procedural management. An in-depth discussion of administration of tumescent local anesthesia and proper liposuction technique is also included. Lastly, a number of ancillary techniques to further enhance the appearance of the neck including laser lipolysis, fractional ablative CO(2) resurfacing, and treatment of platysmal banding will be briefly discussed. © 2011 Wiley Periodicals, Inc.

  20. 77 FR 40413 - Third-Party Provision of Ancillary Services; Accounting and Financial Reporting for New Electric...

    Science.gov (United States)

    2012-07-09

    ... Ancillary Services; Accounting and Financial Reporting for New Electric Storage Technologies AGENCY: Federal... Ancillary Services; Accounting and Financial Reporting for New Electric Storage Technologies, 135 FERC ] 61... Commission 18 CFR Parts 35, 37, and 101 Third-Party Provision of Ancillary Services; Accounting and Financial...

  1. Exploiting ancillary services from distributed generation - perspectives for the Danish power system

    DEFF Research Database (Denmark)

    Nyeng, Preben; Pedersen, Knud Ole Helgesen; Østergaard, Jacob

    2007-01-01

    The share of the electric power production originating from distributed energy resources has rapidly increased during the recent past. However when it comes to ancillary services necessary to ensure the stability and appropriate operation of the power system, the distributed energy resources take...... a very passive role. This paper outlines suggestions on how to activate the potential of ancillary services from distributed energy resources, thereby exploiting their ability to contribute to power system operation. Furthermore, methods for integrating the ancillary service delivery into a deregulated...... power system are proposed and evaluated....

  2. Commercial Building Loads Providing Ancillary Services in PJM

    Energy Technology Data Exchange (ETDEWEB)

    MacDonald, Jason; Kiliccote, Sila; Boch, Jim; Chen, Jonathan; Nawy, Robert

    2014-06-27

    The adoption of low carbon energy technologies such as variable renewable energy and electric vehicles, coupled with the efficacy of energy efficiency to reduce traditional base load has increased the uncertainty inherent in the net load shape. Handling this variability with slower, traditional resources leads to inefficient system dispatch, and in some cases may compromise reliability. Grid operators are looking to future energy technologies, such as automated demand response (DR), to provide capacity-based reliability services as the need for these services increase. While DR resources are expected to have the flexibility characteristics operators are looking for, demonstrations are necessary to build confidence in their capabilities. Additionally, building owners are uncertain of the monetary value and operational burden of providing these services. To address this, the present study demonstrates the ability of demand response resources providing two ancillary services in the PJM territory, synchronous reserve and regulation, using an OpenADR 2.0b signaling architecture. The loads under control include HVAC and lighting at a big box retail store and variable frequency fan loads. The study examines performance characteristics of the resource: the speed of response, communications latencies in the architecture, and accuracy of response. It also examines the frequency and duration of events and the value in the marketplace which can be used to examine if the opportunity is sufficient to entice building owners to participate.

  3. Survey of U.S. Ancillary Services Markets

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Zhi; Levin, Todd; Conzelmann, Guenter

    2016-06-01

    In addition to providing energy to end-consumers, power system operators are also responsible for ensuring system reliability. To this end, power markets maintain an array of ancillary services to ensure that it is always possible to balance the supply and demand for energy in real-time. A subset of these ancillary services are commonly procured through market-based mechanisms: namely, Regulation, Spinning, and Non-spinning Reserves. Regulation Reserves are maintained to respond to supply/demand imbalances over short time frames, typically on the order of several seconds to one minute. Resources that provide Regulation Reserves adjust their generation or load levels in response to automatic generation control (AGC) signals provided by the system operator. Contingency reserves are maintained to provide additional generation capacity in the event that load increases substantially or supply side resources reduce their output or are taken offline. The reserves are typically segmented into two categories, 1) Spinning or Synchronized Reserves that are provided by generation units that are actively generating and have the ability to increase or decrease their output, 2) Non-spinning or Non-synchronized Reserves that are provided by generation resources that are not actively generating, but are able to start up and provide generation within a specified timeframe. Contingency reserves typically have response times on the order of ten to 30 minutes and can also be provided by demand-side resources that are capable of reducing their load. There are seven distinct power markets in the United States, each operated by a Regional Transmission Operator (RTO) or Independent System Operator (ISO) that operates the transmission system in its territory, operates markets for energy and ancillary services, and maintains system reliability. Each power market offers its own set of ancillary services, and precise definitions, requirements, and market mechanisms differ between markets

  4. The beta subunit of casein kinase II

    DEFF Research Database (Denmark)

    Boldyreff, B; Piontek, K; Schmidt-Spaniol, I

    1991-01-01

    cDNAs encoding the beta subunit of pig and mouse CKII were isolated. The porcine cDNA was expressed as a fusion protein in Escherichia coli and used for the production of anti-CKII-beta subunit specific antibodies.......cDNAs encoding the beta subunit of pig and mouse CKII were isolated. The porcine cDNA was expressed as a fusion protein in Escherichia coli and used for the production of anti-CKII-beta subunit specific antibodies....

  5. EX1504L4 Dive03 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L4: Campaign to Address...

  6. EX1504L3 Dive07 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L3: CAPSTONE Leg III:...

  7. EX1605L3 Dive04 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  8. EX1606 Dive03 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1606: CAPSTONE Wake Island...

  9. EX1504L4 Dive02 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L4: Campaign to Address...

  10. EX1606 Dive01 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1606: CAPSTONE Wake Island...

  11. EX1606 Dive09 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1606: CAPSTONE Wake Island...

  12. EX1605L3 Dive01 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  13. EX1606 Dive14 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1606: CAPSTONE Wake Island...

  14. EX1504L3 Dive05 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L3: CAPSTONE Leg III:...

  15. EX1605L3 Dive20 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  16. EX1504L2 Dive14 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L2: Campaign to Address...

  17. EX1606 Dive10 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1606: CAPSTONE Wake Island...

  18. EX1504L4 Dive12 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L4: Campaign to Address...

  19. EX1504L3 Dive03 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L3: CAPSTONE Leg III:...

  20. EX1605L3 Dive15 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  1. EX1605L3 Dive09 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  2. EX1605L3 Dive03 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  3. EX1605L3 Dive13 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  4. EX1504L2 Dive05 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L2: Campaign to Address...

  5. EX1605L3 Dive02 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  6. EX1606 Dive07 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1606: CAPSTONE Wake Island...

  7. EX1504L4 Dive06 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L4: Campaign to Address...

  8. EX1504L2 Dive18 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L2: Campaign to Address...

  9. EX1606 Dive11 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1606: CAPSTONE Wake Island...

  10. EX1605L3 Dive08 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  11. EX1504L2 Dive16 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L2: Campaign to Address...

  12. EX1605L3 Dive10 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  13. EX1605L3 Dive16 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  14. EX1606 Dive08 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1606: CAPSTONE Wake Island...

  15. EX1605L3 Dive22 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  16. EX1605L3 Dive17 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  17. EX1606 Dive13 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1606: CAPSTONE Wake Island...

  18. EX1504L4 Dive10 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L4: Campaign to Address...

  19. EX1605L3 Dive06 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  20. EX1605L3 Dive12 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  1. EX1504L3 Dive06 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L3: CAPSTONE Leg III:...

  2. EX1606 Dive12 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1606: CAPSTONE Wake Island...

  3. EX1605L3 Dive05 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  4. EX1605L3 Dive21 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  5. EX1606 Dive04 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1606: CAPSTONE Wake Island...

  6. EX1605L3 Dive19 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  7. EX1605L3 Dive14 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  8. EX1504L2 Dive12 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L2: Campaign to Address...

  9. EX1605L3 Dive11 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  10. EX1605L3 Dive07 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1605L3: CAPSTONE CNMI &...

  11. EX1504L2 Dive06 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L2: Campaign to Address...

  12. EX1606 Dive06 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1606: CAPSTONE Wake Island...

  13. EX1606 Dive02 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1606: CAPSTONE Wake Island...

  14. EX1606 Dive05 Ancillary Data Collection including reports, kmls, spreadsheets, and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1606: CAPSTONE Wake Island...

  15. Trace Metal and Ancillary Data in Puget Sound, 1980 - 1986 (NODC Accession 9100153)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — In first of three data reports on the trace metal and ancillary data in Puget Sound and its watershed (Paulson et al., 1991a), all water column, sediment, and...

  16. EX1504L2 Dive04 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L2: Campaign to Address...

  17. EX1504L2 Dive01 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L2: Campaign to Address...

  18. NACP Site: Tower Meteorology, Flux Observations with Uncertainty, and Ancillary Data

    Data.gov (United States)

    National Aeronautics and Space Administration — ABSTRACT: This data set contains meteorological, carbon cycle flux, phenology, and ancillary data measured at 47 eddy covariance flux tower sites across North...

  19. EX1504L3 Dive04 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L3: CAPSTONE Leg III:...

  20. EX1504L2 Dive17 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L2: Campaign to Address...

  1. EX1504L2 Dive02 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L2: Campaign to Address...

  2. EX1504L3 Dive02 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L3: CAPSTONE Leg III:...

  3. EX1504L4 Dive04 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L4: Campaign to Address...

  4. EX1504L2 Dive08 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L2: Campaign to Address...

  5. EX1504L2 Dive10 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L2: Campaign to Address...

  6. EX1504L2 Dive13 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L2: Campaign to Address...

  7. NACP Site: Tower Meteorology, Flux Observations with Uncertainty, and Ancillary Data

    Data.gov (United States)

    National Aeronautics and Space Administration — This data set contains meteorological, carbon cycle flux, phenology, and ancillary data measured at 47 eddy covariance flux tower sites across North America. The...

  8. Ocean Surface Topography Mission (OSTM) /Jason-2: Ancillary Files (NODC Accession 0044982)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This accession contains the data descriptions for the OSTM/Jason-2 Ancillary data files, which is served through the NOAA/NESDIS Comprehensive Large Array-data...

  9. EX1504L4 Dive01 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L4: Campaign to Address...

  10. EX1504L4 Dive05 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L4: Campaign to Address...

  11. EX1504L2 Dive11 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L2: Campaign to Address...

  12. EX1504L4 Dive09 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L4: Campaign to Address...

  13. EX1504L4 Dive11 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L4: Campaign to Address...

  14. EX1504L2 Dive09 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L2: Campaign to Address...

  15. EX1504L4 Dive13 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L4: Campaign to Address...

  16. EX1504L4 Dive08 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L4: Campaign to Address...

  17. EX1504L2 Dive07 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L2: Campaign to Address...

  18. EX1504L4 Dive07 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L4: Campaign to Address...

  19. EX1504L2 Dive15 Ancillary Data Collection including reports, kmls, spreadsheets, images and data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Standard suite of ancillary data files generated through a scripting process following an ROV dive on NOAA Ship Okeanos Explorer during EX1504L2: Campaign to Address...

  20. Performance Requirements Modeling andAssessment for Active Power Ancillary Services

    DEFF Research Database (Denmark)

    Bondy, Daniel Esteban Morales; Thavlov, Anders; Tougaard, Janus Bundsgaard Mosbæk

    2017-01-01

    system operation, a reliable service delivery is required, yet it may not be appropriate to apply conventional performance requirements to new technologies and methods. The service performance requirements and assessment methods therefore need to be generalized and standardized in order to include future...... ancillary service sources. This paper develops a modeling method for ancillary services performance requirements, including performance and verification indices. The use of the modeling method and the indices is exemplified in two case studies....

  1. Ancillary benefits of climate policy in a small open economy: The case of Sweden

    International Nuclear Information System (INIS)

    Krook Riekkola, Anna; Ahlgren, Erik O.; Soederholm, Patrik

    2011-01-01

    It is increasingly recognised that GHG reduction policies can have important ancillary benefits in the form of positive local and regional environmental impacts. The purpose of this paper is to estimate the domestic ancillary pollution benefits of climate policy in Sweden, and investigate how these are affected by different climate policy designs. The latter differ primarily in terms of how the country chooses to meet a specific target and where the necessary emission reductions take place. The analysis relies on simulations within the energy system optimisation model TIMES-Sweden, and focuses on four non-GHG pollutants: Nitrogen Oxides (NO X ), Non Methane Volatile Organic Compounds (NMVOC), inhalable particles (PM 2.5 ), and Sulphur dioxide (SO 2 ). The simulations permit detailed assessments of the respective technology and fuel choices that underlie any net changes in the estimated ancillary effects. The results indicate that the ancillary benefits constitute a far from insignificant share of total system costs, and this share appears to be highest in the scenarios that entail the largest emission reductions domestically. This result reflects the fact that carbon dioxide emission reductions abroad also implies a lost opportunity of achieving substantial domestic welfare gain from the reductions of regional and local environmental pollutants. - Highlights: → We estimate the domestic ancillary pollution benefits of climate policy in Sweden. → These constitute a sizeable share of total system costs. → The ancillary benefits are highest in the policy scenarios that entail the largest emission reductions domestically.

  2. Subunit heterogeneity in the lima bean lectin.

    Science.gov (United States)

    Roberts, D D; Etzler, M E; Goldstein, I J

    1982-08-10

    Three forms of lectin (components I, II, and III) from lima beans (Phaseolus lunatus) have been purified on an affinity support containing the synthetic type A blood group trisaccharide alpha-D-GalNAc-(1 leads to 3)-[alpha-L-Fuc-(1 leads to 2)]-beta-D-Gal-(1 leads to). Conversion of components I and II to component III has been achieved by reduction in 10(-2) M dithiothreitol. Isoelectric focusing of lima bean lectin in the presence of 8 M urea and beta-mercaptoethanol revealed charge heterogeneity of the lectin subunits. Three major subunit classes of apparent pI 7.05, 6.65, and 6.45, designated alpha, beta, and alpha', respectively, were identified; they occur in a relative abundance of 2:5:3. Green lima beans harvested before maturity lacked the alpha' subunit (pI 6.45) which appears to accumulate during seed maturation. The three subunits are glycoproteins of identical size and immunochemical reactivity. Identical NH2-terminal sequences were found for the three subunits. Amino acid analysis and tryptic peptide mapping indicated that the observed charge heterogeneity is probably due to differences in the primary structure of the subunits. Studies of subunit composition of charge isolectins provided evidence of nonrandom subunit assembly. A model is proposed involving pairing of a pI 6.65 subunit with either a pI 7.06 or 6.45 subunit to form dimeric units. Possible roles for subunit heterogeneity and ordered subunit assembly in determining the metal and sugar binding properties of lima bean lectin are discussed.

  3. More than a score: a qualitative study of ancillary benefits of performance measurement.

    Science.gov (United States)

    Powell, Adam A; White, Katie M; Partin, Melissa R; Halek, Krysten; Hysong, Sylvia J; Zarling, Edwin; Kirsh, Susan R; Bloomfield, Hanna E

    2014-08-01

    Prior research has examined clinical effects of performance measurement systems. To the extent that non-clinical effects have been researched, the focus has been on negative unintended consequences. Yet, these same systems may also have ancillary benefits for patients and providers--that is, benefits that extend beyond improvements on clinical measures. The purpose of this study is to identify and describe potential ancillary benefits of performance measures as perceived by primary care staff and facility leaders in a large US healthcare system. In-person individual semistructured interviews were conducted with 59 primary care staff and facility leaders at four Veterans Health Administration facilities. Transcribed interviews were coded and organised into thematic categories. Interviewed staff observed that local performance measurement implementation practices can result in increased patient knowledge and motivation. These effects on patients can lead to improved performance scores and additional ancillary benefits. Performance measurement implementation can also directly result in ancillary benefits for the patients and providers. Patients may experience greater satisfaction with care and psychosocial benefits associated with increased provider-patient communication. Ancillary benefits of performance measurement for providers include increased pride in individual or organisational performance and greater confidence that one's practice is grounded in evidence-based medicine. A comprehensive understanding of the effects of performance measurement systems needs to incorporate ancillary benefits as well as effects on clinical performance scores and negative unintended consequences. Although clinical performance has been the focus of most evaluations of performance measurement to date, both patient care and provider satisfaction may improve more rapidly if all three categories of effects are considered when designing and evaluating performance measurement systems

  4. Role of the Rubisco Small Subunit

    Energy Technology Data Exchange (ETDEWEB)

    Spreitzer, Robert Joseph [Univ. of Nebraska, Lincoln, NE (United States)

    2016-11-05

    Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyzes the rate-limiting step of CO2 fixation in photosynthesis. However, it is a slow enzyme, and O2 competes with CO2 at the active site. Oxygenation initiates the photorespiratory pathway, which also results in the loss of CO2. If carboxylation could be increased or oxygenation decreased, an increase in net CO2 fixation would be realized. Because Rubisco provides the primary means by which carbon enters all life on earth, there is much interest in engineering Rubisco to increase the production of food and renewable energy. Rubisco is located in the chloroplasts of plants, and it is comprised of two subunits. Much is known about the chloroplast-gene-encoded large subunit (rbcL gene), which contains the active site, but much less is known about the role of the nuclear-gene-encoded small subunit in Rubisco function (rbcS gene). Both subunits are coded by multiple genes in plants, which makes genetic engineering difficult. In the eukaryotic, green alga Chlamydomonas reinhardtii, it has been possible to eliminate all the Rubisco genes. These Rubisco-less mutants can be maintained by providing acetate as an alternative carbon source. In this project, focus has been placed on determining whether the small subunit might be a better genetic-engineering target for improving Rubisco. Analysis of a variable-loop structure (βA-βB loop) of the small subunit by genetic selection, directed mutagenesis, and construction of chimeras has shown that the small subunit can influence CO2/O2 specificity. X-ray crystal structures of engineered chimeric-loop enzymes have indicated that additional residues and regions of the small subunit may also contribute to Rubisco function. Structural dynamics of the small-subunit carboxyl terminus was also investigated. Alanine-scanning mutagenesis of the most-conserved small-subunit residues has identified a

  5. Loads Providing Ancillary Services: Review of InternationalExperience-- Technical Appendix: Market Descriptions

    Energy Technology Data Exchange (ETDEWEB)

    Grayson Heffner, Charles Goldman, Kintner-Meyer, M; Kirby, Brendan

    2007-05-01

    In this study, we examine the arrangements for andexperiences of end-use loads providing ancillary services (AS) in fiveelectricity markets: Australia, the United Kingdom (UK), the Nordicmarket, and the ERCOT and PJM markets in the United States. Our objectivein undertaking this review of international experience was to identifyspecific approaches or market designs that have enabled customer loads toeffectively deliver various ancillary services (AS) products. We hopethat this report will contribute to the ongoing discussion in the U.S.and elsewhere regarding what institutional and technical developments areneeded to ensure that customer loads can meaningfully participate in allwholesale electricity markets.

  6. 28 CFR 51.6 - Political subunits.

    Science.gov (United States)

    2010-07-01

    ... 28 Judicial Administration 2 2010-07-01 2010-07-01 false Political subunits. 51.6 Section 51.6 Judicial Administration DEPARTMENT OF JUSTICE (CONTINUED) PROCEDURES FOR THE ADMINISTRATION OF SECTION 5 OF THE VOTING RIGHTS ACT OF 1965, AS AMENDED General Provisions § 51.6 Political subunits. All political...

  7. Binding of CFA/I Pili of Enterotoxigenic Escherichia coli to Asialo-GM1 Is Mediated by the Minor Pilin CfaE.

    Science.gov (United States)

    Madhavan, T P Vipin; Riches, James D; Scanlon, Martin J; Ulett, Glen C; Sakellaris, Harry

    2016-05-01

    CFA/I pili are representatives of a large family of related pili that mediate the adherence of enterotoxigenic Escherichia coli to intestinal epithelial cells. They are assembled via the alternate chaperone-usher pathway and consist of two subunits, CfaB, which makes up the pilus shaft and a single pilus tip-associated subunit, CfaE. The current model of pilus-mediated adherence proposes that CFA/I has two distinct binding activities; the CfaE subunit is responsible for binding to receptors of unknown structure on erythrocyte and intestinal epithelial cell surfaces, while CfaB binds to various glycosphingolipids, including asialo-GM1. In this report, we present two independent lines of evidence that, contrary to the existing model, CfaB does not bind to asialo-GM1 independently of CfaE. Neither purified CfaB subunits nor CfaB assembled into pili bind to asialo-GM1. Instead, we demonstrate that binding activity toward asialo-GM1 resides in CfaE and this is essential for pilus binding to Caco-2 intestinal epithelial cells. We conclude that the binding activities of CFA/I pili for asialo-GM1, erythrocytes, and intestinal cells are inseparable, require the same amino acid residues in CfaE, and therefore depend on the same or very similar binding mechanisms. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  8. 78 FR 46177 - Third-Party Provision of Ancillary Services; Accounting and Financial Reporting for New Electric...

    Science.gov (United States)

    2013-07-30

    ...-Party Provision of Ancillary Services; Accounting and Financial Reporting for New Electric Storage... Financial Reporting for New Electric Storage Technologies, Notice of Proposed Rulemaking, FERC Stats. & Regs... Commission 18 CFR Parts 35 and 101 Third-Party Provision of Ancillary Services; Accounting and Financial...

  9. 29 CFR 2550.408b-6 - Statutory exemption for ancillary services by a bank or similar financial institution.

    Science.gov (United States)

    2010-07-01

    ....408b-6 Statutory exemption for ancillary services by a bank or similar financial institution. (a) In... service is consistent with sound banking and financial practice, as determined by Federal or State... 29 Labor 9 2010-07-01 2010-07-01 false Statutory exemption for ancillary services by a bank or...

  10. Analysis of the Impact of Wind Power Participating in Both Energy and Ancillary Services Markets – The Danish Case

    DEFF Research Database (Denmark)

    Soares, Tiago; Morais, Hugo; Pinson, Pierre

    2014-01-01

    strategies that wind farms employ on ene rgy and ancillary service market. A lready proposed Proportional Wind Reserve Strategy (PWRS) and a Continuo us Wind Reserve Strategy (CWRS) are used to determine the amount of available power for ancillary services. A case study based on real and recent dat...

  11. Mixed integer non-linear programming and Artificial Neural Network based approach to ancillary services dispatch in competitive electricity markets

    International Nuclear Information System (INIS)

    Canizes, Bruno; Soares, João; Faria, Pedro; Vale, Zita

    2013-01-01

    Highlights: • Ancillary services market management. • Ancillary services requirements forecast based on Artificial Neural Network. • Ancillary services clearing mechanisms without complex bids and with complex bids. - Abstract: Ancillary services represent a good business opportunity that must be considered by market players. This paper presents a new methodology for ancillary services market dispatch. The method considers the bids submitted to the market and includes a market clearing mechanism based on deterministic optimization. An Artificial Neural Network is used for day-ahead prediction of Regulation Down, regulation-up, Spin Reserve and Non-Spin Reserve requirements. Two test cases based on California Independent System Operator data concerning dispatch of Regulation Down, Regulation Up, Spin Reserve and Non-Spin Reserve services are included in this paper to illustrate the application of the proposed method: (1) dispatch considering simple bids; (2) dispatch considering complex bids

  12. 17 CFR 240.3b-15 - Definition of ancillary portfolio management securities activities.

    Science.gov (United States)

    2010-04-01

    ... 17 Commodity and Securities Exchanges 3 2010-04-01 2010-04-01 false Definition of ancillary portfolio management securities activities. 240.3b-15 Section 240.3b-15 Commodity and Securities Exchanges... Rules and Regulations Under the Securities Exchange Act of 1934 Definitions § 240.3b-15 Definition of...

  13. Regulatory issues associated with closure of the Hanford AX Tank Farm ancillary equipment

    International Nuclear Information System (INIS)

    Becker, D.L.

    1998-01-01

    Liquid mixed, high-level radioactive waste has been stored in underground single-shell tanks at the US Department of Energy's (DOE's) Hanford Site. After retrieval of the waste from the single-shell tanks, the DOE will proceed with closure of the tank farm. The 241-AX Tank Farm includes four one-million gallon single-shell tanks in addition to sluice lines, transfer lines, ventilation headers, risers, pits, cribs, catch tanks, buildings, well and associated buried piping. This equipment is classified as ancillary equipment. This document addresses the requirements for regulatory close of the ancillary equipment in the Hanford Site 241-AX Tank Farm. The options identified for physical closure of the ancillary equipment include disposal in place, disposal in place after treatment, excavation and disposal on site in an empty single-shell tank, and excavation and disposal outside the AX Tank Farm. The document addresses the background of the Hanford Site and ancillary equipment in the AX Tank Farm, regulations for decontamination and decommissioning of radioactively contaminated equipment, requirements for the cleanup and disposal of radioactive wastes, cleanup and disposal requirements governing hazardous and mixed waste, and regulatory requirements and issues associated with each of the four physical closure options. This investigation was conducted by the Sandia National Laboratories, Albuquerque, New Mexico, during Fiscal Year 1998 for the Hanford Tanks Initiative Project

  14. 30 CFR 250.207 - What ancillary activities may I conduct?

    Science.gov (United States)

    2010-07-01

    ... surveys; or (c) Studies that model potential oil and hazardous substance spills, drilling muds and... 30 Mineral Resources 2 2010-07-01 2010-07-01 false What ancillary activities may I conduct? 250.207 Section 250.207 Mineral Resources MINERALS MANAGEMENT SERVICE, DEPARTMENT OF THE INTERIOR OFFSHORE...

  15. Technical Feasibility of Ancillary Services provided by ReGen Plants

    DEFF Research Database (Denmark)

    Altin, Müfit; Han, Xue; Hansen, Anca Daniela

    This report is the first deliverable in WP1 in the project “Ancillary services from renewable power plants” (RePlan). RePlan is funded as PSO project 2015 no. 12347 by the Danish PSO-programme ForskEL, which is administered by Energinet.DK. RePlan is carried out in collaboration between DTU Wind ...

  16. Optimal Sizing and Allocation of Residential Photovoltaic Panels in a Distribution Network for Ancillary Services Application

    DEFF Research Database (Denmark)

    Kordheili, Reza Ahmadi; Pourmousavi, Ali; Pillai, Jayakrishnan Radhakrishna

    2014-01-01

    requirements, such as voltage and current limits. This paper proposes an optimization method for determining the number of photovoltaic (PV) panels together with their arrangement in the grid in order to maximize ancillary service, without violating grid operation limits. The proposed optimization method...

  17. Ethics of Ancillary Care in Clinical Trials in Low Income Countries: A ...

    African Journals Online (AJOL)

    These limits should be negotiated to ensure that the research remains feasible and economically viable. (Afr J Reprod Health 2014; 18[3]: 135-142). Keywords: Ancillary care; ethics, HIV prevention, undue inducement. La conduite éthique des chercheurs dans le domaine de la prévention du VIH est soumise au contrôle.

  18. Ancillary obligations as an additional obligation of a limited liability company's member

    Directory of Open Access Journals (Sweden)

    Marjanski Vladimir Ž.

    2016-01-01

    Full Text Available Ancillary obligation can be defined as an obligation of a limited liability company's member undertaken through the instrument of incorporation, which can take the form of monetary or non-monetary obligation (usually it is an obligation of a non-monetary character, which has a certain financial value and can be the object of a legally valid obligation. Ancillary obligations, as a contribution to activities of the company, are not regulated in the Law on Companies. The legal nature of this obligation is different from the obligation to make a contribution in money or in kind to a company's assets, to make additional pay-ins or from the landing of funds to the company. Ancillary obligation is an optional and additional obligation of a limited liability company's member and the obligation itself and its contents is defined in the instrument of incorporation. When a limited liability company's member undertakes an ancillary obligation this creates a distinctive relationship between the member and the company, concerning their respective obligations and the legal status of the company, as well as certain consequences derived from those obligations.

  19. Ancillary services: technical specifications, system needs and costs. Deliverable D 2.2

    DEFF Research Database (Denmark)

    Holttinen, Hannele; Cutululis, Nicolaos Antonio; Gubina, Andrej

    2012-01-01

    In this report, different ancillary services are described and a table listing main services is presented. While Chapter 2 is describing the services from (renewable) generators point of view, Chapter 3 is considering future system needs for services with increased wind and solar penetration...

  20. Acetylcholine Receptor: Complex of Homologous Subunits

    Science.gov (United States)

    Raftery, Michael A.; Hunkapiller, Michael W.; Strader, Catherine D.; Hood, Leroy E.

    1980-06-01

    The acetylcholine receptor from the electric ray Torpedo californica is composed of five subunits; two are identical and the other three are structurally related to them. Microsequence analysis of the four polypeptides demonstrates amino acid homology among the subunits. Further sequence analysis of both membrane-bound and Triton-solubilized, chromatographically purified receptor gave the stoichiometry of the four subunits (40,000:50,000:60,000:65,000 daltons) as 2:1:1:1, indicating that this protein is a pentameric complex with a molecular weight of 255,000 daltons. Genealogical analysis suggests that divergence from a common ancestral gene occurred early in the evolution of the receptor. This shared ancestry argues that each of the four subunits plays a functional role in the receptor's physiological action.

  1. Risk capital allocation with autonomous subunits

    DEFF Research Database (Denmark)

    Hougaard, Jens Leth; Smilgins, Aleksandrs

    2016-01-01

    Risk capital allocation problems have been widely discussed in the academic literature. We consider a set of independent subunits collaborating in order to reduce risk: that is, when subunit portfolios are merged a diversification benefit arises and the risk of the group as a whole is smaller tha...... fairness tests related directly to the problem of risk capital allocation and show that the Lorenz set satisfies all three tests in contrast to other well-known coherent methods. Finally, we discuss how to deal with non-uniqueness of the Lorenz set.......Risk capital allocation problems have been widely discussed in the academic literature. We consider a set of independent subunits collaborating in order to reduce risk: that is, when subunit portfolios are merged a diversification benefit arises and the risk of the group as a whole is smaller than...... the sum of the risks of the individual subunits. The question is how to allocate the risk capital of the group among the subunits in a fair way. In this paper we propose to use the Lorenz set as an allocation method. We show that the Lorenz set is operational and coherent. Moreover, we propose three...

  2. Influence of Wind Plant Ancillary Frequency Control on System Small Signal Stability

    DEFF Research Database (Denmark)

    Su, Chi; Chen, Zhe

    2012-01-01

    Since large-scale wind farms are increasingly connected to modern power grids, the transmission system operators put more requirements as part of the grid codes on the integration of wind farms. System frequency control which is normally provided by conventional synchronous generators becomes...... a common requirement to wind farms. This ancillary frequency control provided by wind farms could have some influence on the system small signal stability. This paper implements an ancillary frequency control strategy on a directdrive- full-convertor-based wind farm and studies its influence on the damping...... ratio values of the dominant oscillation modes within the connected power system. All the calculations and simulations are conducted in DIgSILENT PowerFactory 14.0....

  3. Influence of Wind Plant Ancillary Voltage Control on System Small Signal Stability

    DEFF Research Database (Denmark)

    Su, Chi; Chen, Zhe

    2012-01-01

    As a common tendency, large-scale wind farms are increasingly connected to the transmission system of modern power grids. This introduces some new challenges to the connected power systems, and the transmission system operators (TSOs) have to put some new requirements as part of the grid codes...... on the integration of wind farms. One common requirement to wind farms is the function of system voltage control which can be implemented in the grid-side convertor controller of a variable speed wind turbine. This ancillary voltage control provided by wind farms could have some influence on the system small signal...... stability. This paper implements an ancillary voltage control strategy on a direct-drive-full-convertor-based wind farm and studies its influence on the damping ratio values of the dominant oscillation mode within the connected power system. All the calculations and simulations are conducted in DIg...

  4. Market and policy barriers for demand response providing ancillary services in U.S. markets

    Energy Technology Data Exchange (ETDEWEB)

    Cappers, Peter [Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States); MacDonald, Jason [Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States); Goldman, Charles [Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

    2013-03-01

    This study provides an examination of various market and policy barriers to demand response providing ancillary services in both ISO/RTO and non-ISO/RTO regions, especially at the program provider level. It is useful to classify barriers in order to create a holistic understanding and identify parties that could be responsible for their removal. This study develops a typology of barriers focusing on smaller customers that must rely on a program provider (i.e., electric investor owned utility or IOU, ARC) to create an aggregated DR resource in order to bring ancillary services to the balancing authority. The barriers were identified through examinations of regulatory structures, market environments, and product offerings; and discussions with industry stakeholders and regulators. In order to help illustrate the differences in barriers among various wholesale market designs and their constituent retail environments, four regions were chosen to use as case studies: Colorado, Texas, Wisconsin, and New Jersey.

  5. VOLTTRON-Based System for Providing Ancillary Services with Residential Building Loads

    Energy Technology Data Exchange (ETDEWEB)

    Jin, Xin

    2016-07-01

    Ancillary services entail controlled modulation of building equipment to maintain a stable balance of generation and load in the power system. Ancillary services include frequency regulation and contingency reserves, whose acting time ranges from several seconds to several minutes. Many pilot studies have been implemented to use industrial loads to provide ancillary services, and some have explored services from commercial building loads or electric vehicle charging loads. Residential loads, such as space conditioning and water heating, represent a largely untapped resource for providing ancillary services. The residential building sector accounts for a significant fraction of the total electricity use in the United States. Many loads in residential buildings are flexible and could potentially be curtailed or shifted at the request of the grid. However, there are many barriers that prevent residential loads being widely used for ancillary services. One of the major technical barriers is the lack of communication capabilities between end-use devices and the grid. End-use devices need to be able to receive the automatic generation control (AGC) signal from the grid operator and supply certain types of telemetry to verify response. With the advance of consumer electronics, communication-enabled, or 'connected,' residential equipment has emerged to overcome the communication barrier. However, these end-use devices have introduced a new interoperability challenge due to the existence of numerous standards and communication protocols among different end devices. In this paper, we present a VOLTTRON-based system that overcomes these technical challenges and provides ancillary services with residential loads. VOLTTRON is an open-source control and sensing platform for building energy management, facilitating interoperability solutions for end devices. We have developed drivers to communicate and control different types of end devices through standard

  6. Grid-Scale Energy Storage Demonstration of Ancillary Services Using the UltraBattery Technology

    Energy Technology Data Exchange (ETDEWEB)

    Seasholtz, Jeff [East Penn Mfg. Co., Inc., Lyons, PA (United States)

    2015-08-20

    The collaboration described in this document is being done as part of a cooperative research agreement under the Department of Energy’s Smart Grid Demonstration Program. This document represents the Final Technical Performance Report, from July 2012 through April 2015, for the East Penn Manufacturing Smart Grid Program demonstration project. This Smart Grid Demonstration project demonstrates Distributed Energy Storage for Grid Support, in particular the economic and technical viability of a grid-scale, advanced energy storage system using UltraBattery ® technology for frequency regulation ancillary services and demand management services. This project entailed the construction of a dedicated facility on the East Penn campus in Lyon Station, PA that is being used as a working demonstration to provide regulation ancillary services to PJM and demand management services to Metropolitan Edison (Met-Ed).

  7. Ancillary Frequency Control of Direct Drive Full-Scale Converter Based Wind Power Plants

    DEFF Research Database (Denmark)

    Hu, Weihao; Su, Chi; Fang, Jiakun

    2013-01-01

    emulation, primary frequency control and secondary frequency control, are proposed in order to improve the frequency stability of power systems. The modified IEEE 39-bus test system with a large-scale wind power penetration is chosen as the studied power system. Simulation results show that the proposed......This paper presents a simulation model of a wind power plant based on a MW-level variable speed wind turbine with a full-scale back-to-back power converter developed in the simulation tool of DIgSILENT Power Factory. Three different kinds of ancillary frequency control strategies, namely inertia...... control strategies are effective means for providing ancillary frequency control of variable speed wind turbines with full-scale back-to-back power converters....

  8. The State of Cell Blocks and Ancillary Testing: Past, Present, and Future.

    Science.gov (United States)

    Saqi, Anjali

    2016-12-01

    Cell blocks are an integral part of cytology, but their utility is recognized probably more now than ever before, largely owing to the significant role they play in ancillary testing, particularly molecular diagnostics. Modifications to improve the cell block method initially introduced more than a century ago have been made over the years. Though their value is acknowledged and they are widely used across laboratories, cell block preparations are not standardized and results of ancillary testing performed on them are inconsistent. This article reviews the state of cell blocks-summarizes the more common, currently available and used methods and their corresponding advantages and shortcomings, outlines the role of alternative techniques (eg, smears), and proposes methods to optimize results.

  9. The TIGRESS Integrated Plunger ancillary systems for electromagnetic transition rate studies at TRIUMF

    International Nuclear Information System (INIS)

    Voss, P.; Henderson, R.; Andreoiu, C.; Ashley, R.; Austin, R.A.E.; Ball, G.C.; Bender, P.C.; Bey, A.; Cheeseman, A.; Chester, A.; Cross, D.S.; Drake, T.E.; Garnsworthy, A.B.; Hackman, G.; Holland, R.; Ketelhut, S.; Kowalski, P.; Krücken, R.; Laffoley, A.T.; Leach, K.G.

    2014-01-01

    The TIGRESS Integrated Plunger device is a new experimental tool for nuclear structure investigations via gamma-ray spectroscopy with post-accelerated beams from the ISAC-II facility at TRIUMF. Several ancillary detection systems integral to the device's capabilities for charged-particle tagging and light-ion identification following a variety of nuclear reaction mechanisms have been constructed and characterized. In particular, a silicon PIN diode wall, an annular silicon segmented detector, and a CsI(Tl) scintillator wall have together enabled particle-gamma correlations for reaction channel selectivity and precision kinematic reconstruction in recent measurements. We highlight the construction, characteristics, and implementation of the device's ancillary detectors as they enable a rich set of electromagnetic transition rate measurements via Doppler-shift lifetime techniques and low-energy Coulomb excitation

  10. Vertical Keiretsu and International Market Entry: The Case of the Japanese Automobile Ancillary Industry

    OpenAIRE

    Kunal Banerji; Rakesh B Sambharya

    1996-01-01

    This study investigated whether small Japanese automobile ancillary firms followed the major Japanese automakers in setting up production facilities in the United States. Based on Dunning's eclectic theory of foreign direct investment and resource dependence theory, hypotheses were generated to explain the determinants of foreign investment of Japanese suppliers. It was hypothesized that mutual dependencies between the major Japanese automobile firms and their suppliers, firm-specific ownersh...

  11. Application of microgrids in providing ancillary services to the utility grid

    International Nuclear Information System (INIS)

    Majzoobi, Alireza; Khodaei, Amin

    2017-01-01

    A microgrid optimal scheduling model is developed in this paper to demonstrate microgrid's capability in offering ancillary services to the utility grid. The application of localized ancillary services is of significant importance to grid operators as the growing proliferation of distributed renewable energy resources, mainly solar generation, is causing major technical challenges in supply-load balance. The proposed microgrid optimal scheduling model coordinates the microgrid net load with the aggregated consumers/prosumers net load in its connected distribution feeder to capture both inter-hour and intra-hour net load variations. In particular, net load variations for three various time resolutions are considered, including hourly ramping, 10-min based load following, and 1-min based frequency regulation. Numerical simulations on a test distribution feeder with one microgrid and several consumers/prosumers indicate the effectiveness of the proposed model and the viability of the microgrid application in supporting grid operation. - Highlights: • Microgrid optimal scheduling for providing ancillary services to the utility grid. • Local management and mitigation of distribution net load variations. • Offering various support services: ramping, load following, frequency regulation. • Proven effectiveness and accuracy in capturing net load variations.

  12. The Subunit Principle in Scar Face Revision.

    Science.gov (United States)

    Elshahat, Ahmed; Lashin, Riham

    2017-06-01

    Facial scaring is considered one of the most difficult cosmetic problems for any plastic surgeon to solve. The condition is more difficult if the direction of the scar is not parallel to relaxed skin tension lines. Attempts to manage this difficult situation included revisions using geometric designs, Z plasties or W plasties to camouflage the straight line visible scaring. The use of long-lasting resorbable sutures was tried too. Recently, the use of botulinum toxin during revision improved the results. Fractional CO2 lasers, microfat grafts, and platelet-rich plasma were added to the armamentarium. The scar is least visible if placed in the junction between the facial subunits. The aim of this study is to investigate the use of the subunit principle to improve the results of scar revision. Four patients were included in this study. Tissue expansion of the intact part of the subunit allowed shifting the scar to the junction between the affected subunit and the adjacent one. Tissue expansion, delivery of the expanders, and advancement of the flaps were successful in all patients. The fact that this is a 2-stage procedure and sacrifices some of the intact skin from the affected facial subunit, makes this technique reserved to patients with ugly facial scars who are ambitious to improve their appearance.

  13. S3b amino acid substitutions and ancillary subunits alter the affinity of Heteropoda venatoria toxin 2 for Kv4.3.

    Science.gov (United States)

    DeSimone, Christopher V; Lu, YiChun; Bondarenko, Vladimir E; Morales, Michael J

    2009-07-01

    Heteropoda venatoria toxin 2 (HpTx2) is an inhibitor cystine knot (ICK)-gating modifier toxin that selectively inhibits Kv4 channels. To characterize the molecular determinants of interaction, we performed alanine scanning of the Kv4.3 S3b region. HpTx2-Kv4.3 interaction had an apparent K(d) value of 2.3 microM. Two alanine mutants in Kv4.3 increased K(d) values to 6.4 microM for V276A and 25 microM for L275A. Simultaneous mutation of both amino acids to alanine nearly eliminated toxin interaction. Unlike Hanatoxin and other well characterized ICK toxins, HpTx2 binding does not require a charged amino acid for interaction. To determine whether the identity of the S3b binding site amino acids altered HpTx2 specificity, we constructed Kv4.3 [LV275IF]. This mutation decreased the K(d) value to 0.54 microM, suggesting that the hydrophobic character of the putative binding site is the most important property for interaction with HpTx2. One mutant, N280A, caused stronger interaction of HpTx2 with Kv4.3; the K(d) value for Kv4.3 [N280A] was 0.26 microM. To understand Kv4.3-based transient outward currents in native tissues, we tested the affinity of HpTx2 for Kv4.3 coexpressed with KChIP2b. The toxin's K(d) value for Kv4.3 + KChIP2b was 0.95 microM. KChIP2b stabilizes the closed state of Kv4.3, suggesting that the increased toxin affinity is due to increased stabilization of the closed state. These data show that HpTx2 binding to Kv4.3 has aspects in common with other ICK gating modifier toxins but that the interventions that increase toxin affinity suggest flexibility toward channel binding that belies its unusual specificity for Kv4 channels.

  14. Ancillary services analysis of an offshore wind farm cluster-technical integration steps of a simulation tool

    OpenAIRE

    Hennig, Tobias; Löwer, Lothar; Faiella, Luis Mariano; Stock, Sebastian; Jansen, Malte; Hofmann, Lutz; Rohrig, Kurt

    2014-01-01

    In this publication, the authors present methodology and example results for the analysis of ancillary services of an offshore wind farm cluster and its electrical power system. Thereby the operation tool Wind Cluster Management System (WCMS) is used as simulation tool to evaluate certain planning scenarios. Emphasis is made on two topics: 1) the integration of high voltage direct current (HVDC) technology to the WCMS, 2) the ancillary service analysis. As examples, voltage source converter b...

  15. CO{sub 2} mitigation costs and ancillary benefits in the Nordic countries, the UK and Ireland: A survey

    Energy Technology Data Exchange (ETDEWEB)

    Kverndokk, Snorre; Rosendahl, Knut Einar

    2000-07-01

    This paper provides a survey of top-down modelling studies on mitigation costs and ancillary benefits in the Nordic countries, the UK and Ireland. Special emphasis is put on results concerning revenue recycling, double dividend, distributional effects and ancillary benefits. According to the papers surveyed, modest emissions restrictions as those given by the Kyoto Protocol, can be met without substantial costs for the countries studied. (Author)

  16. Is a diagnosis of ancillary restless legs syndrome reproducible over time? Experience with the Wayne Hening telephone diagnostic interview.

    Science.gov (United States)

    Mitterling, Thomas; Frauscher, Birgit; Ehrmann, Laura; Gschliesser, Viola; Brandauer, Elisabeth; Högl, Birgit

    2013-06-01

    The Hening telephone diagnostic interview (HTDI) is a validated structured diagnostic instrument for restless legs syndrome (RLS). A diagnosis of ancillary RLS is defined as RLS with non bothering or only sporadic occurrence of RLS symptoms. The aim of our study was to test the reproducibility of a previously received diagnosis of ancillary RLS with the HTDI in a telephone follow-up examination. Patients with a diagnosis of ancillary RLS underwent a telephone follow-up after an interval of 6 or more months from their entry into the RLS database. The interview included the HTDI, the International RLS Study Group severity rating scale (IRLS) and questions on current RLS medication. Sixty patients with ancillary RLS were eligible for this study, 50 participated. Thirty-six patients were assigned to definite RLS, 11 to probable RLS, one to possible RLS, and two patients were rated as not RLS. Median IRLS score of those with RLS was 10 (0-30). This is the first study to report results of the German version of the HTDI. We demonstrate a high reproducibility of a diagnosis of ancillary RLS over time and that the HTDI is an excellent diagnostic tool for RLS even in ancillary cases. Copyright © 2013 Elsevier B.V. All rights reserved.

  17. Immunochemical aspects of crotoxim and its subunits

    International Nuclear Information System (INIS)

    Nakazone, A.K.

    1979-01-01

    Crotamine and crotoxin with the subunits - phospholipase A and crotapotin - were obtained by purification from Crotalus durissus terrificus venom. Interaction studies of the subunits using crotalic antiserum, indicated that: crotoxin is formed of crotapotin and phospholipase A with the molar ratio of 1 to 1; using crotapotin 125 I the presence of a soluble complex was shown with the same antiserum. Immunological precipitation reactions demonstrated that crotapotin is antigenic: crotapotin and phospholipase A presented similar antigenic determinants; crotoxin antiserum reacted with each one of the submits; when the subunits are mixed to form synthetic crotoxin some antigenic determinants are masked in the process of interaction. Crotamine, interacted with crotapotin 1:1, without hidden antigenic determinants crotapotin antigenic site seems to be formed by, at least, one lysine. Enzimatical activity of phospholipase A apreared to be dependent on some reaction conditions when its arginine residues are blocked. Tyrosines of phospholipase A are more susceptible to labelling with 131 I than crotapotin. Gama irradiation of aqueous solutions of the subunits produced modifications in the ultraviolet spectra. A decrease of the enzymatic activity occured as a function of radiation dosis. Immunological activities of crotapotin and phospholipase A were not altered [pt

  18. Impact of prenatal care provider on the use of ancillary health services during pregnancy

    Science.gov (United States)

    2013-01-01

    Background Recent declines in the provision of prenatal care by family physicians and the integration of midwives into the Canadian health care system have led to a shift in the pattern of prenatal care provision; however it is unknown if this also impacts use of other health services during pregnancy. This study aimed to assess the impact of the type of prenatal care provider on the self-reported use of ancillary services during pregnancy. Methods Data for this study was obtained from the All Our Babies study, a community-based prospective cohort study of women’s experiences during pregnancy and the post-partum period. Chi-square tests and logistic regression were used to assess the association between type of prenatal care provider and use of ancillary health services in pregnancy. Results During pregnancy, 85.8% of women reported accessing ancillary health services. Compared to women who received prenatal care from a family physician, women who saw a midwife were less likely to call a nurse telephone advice line (OR = 0.30, 95% CI: 0.18-0.50) and visit the emergency department (OR = 0.47, 95% CI: 0.24-0.89), but were more likely receive chiropractic care (OR = 4.07, 95% CI: 2.49-6.67). Women who received their prenatal care from an obstetrician were more likely to visit a walk-in clinic (OR = 1.51, 95% CI: 1.11-2.05) than those who were cared for by a family physician. Conclusions Prenatal care is a complex entity and referral pathways between care providers and services are not always clear. This can lead to the provision of fragmented care and create opportunities for errors and loss of information. All types of care providers have a role in addressing the full range of health needs that pregnant women experience. PMID:23497179

  19. Riboprinting of Naegleria spp.: small-subunit versus large-subunit rDNA.

    Science.gov (United States)

    De Jonckheere, J F

    1994-01-01

    The nonpathogenic amoeba Naegleria lovaniensis is closely related to the human pathogen N. fowleri. Both grow at a maximal temperature of 45 degrees C and, therefore, are often found together in the environment. As they are morphologically inseparable at the light-microscope level, refined techniques are necessary to separate the two species. I have used restriction-fragment-length polymorphism analysis of the polymerase chain reaction (PCR)-amplified ribosomal RNA gene, or riboprinting, to distinguish between the different Naegleria spp. Riboprints generated from the small subunit and the large subunit separate N. fowleri from N. lovaniensis. To examine the taxonomic relationships among all Naegleria spp., analysis of the large subunit has to be performed; the small subunit contains a 1.3-kb group I intron, which interferes with tree building based upon restriction sites.

  20. Molecular Structure Laboratory. Fourier Transform Nuclear Magnetic Resonance (FTNMR) Spectrometer and Ancillary Instrumentation at SUNY Geneseo

    Energy Technology Data Exchange (ETDEWEB)

    Geiger, David K [State Univ. of New York (SUNY), Geneseo, NY (United States)

    2015-12-31

    An Agilent 400-MR nuclear magnetic resonance (NMR) spectrometer and ancillary equipment were purchased, which are being used for molecular structure elucidation.  The instrumentation is housed in a pre-existing facility designed specifically for its use. This instrument package is being used to expand the research and educational efforts of the faculty and students at SUNY-Geneseo and is made available to neighboring educational institutions and business concerns.  Funds were also used for training of College personnel, maintenance of the instrumentation, and installation of the equipment.

  1. Enhancing the Role of Electric Vehicles in the Power Grid: Field Validation of Multiple Ancillary Services

    DEFF Research Database (Denmark)

    Knezovic, Katarina; Martinenas, Sergejus; Andersen, Peter Bach

    2016-01-01

    With increased penetration of distributed energy resources and electric vehicles (EVs), different EV integration strategies can be used for mitigating various adverse effects, and supporting the grid. However, the research regarding EV smart charging has mostly remained on simulations, whereas...... essentially means it is applicable to any EV complying with IEC 61851 and SAE J1772 standards. The field test validation is conducted in a real Danish distribution grid with a Nissan Leaf providing three ancillary services through unidirectional AC charging, namely congestion management, local voltage support...

  2. Identification of Conflicts between Transmission and Distribution System Operators when Acquiring Ancillary Services from Electric Vehicles

    DEFF Research Database (Denmark)

    Zecchino, Antonio; Knezovic, Katarina; Marinelli, Mattia

    2017-01-01

    products according to requests coming from both distribution and transmission system operators. The goal of this paper is to provide an identification procedure that is able to detect, identify and catalogue possible conflicts among the involved stakeholders that take place when requesting and/or acquiring...... ancillary services from flexible units. The investigation is carried out considering a 3-area power system which allows to take into account local constraints as well as system-wide needs. As outcome, this paper identifies the conflicts from both a theoretical and a practical point of view, by means...

  3. Involvement of proteasomal subunits zeta and iota in RNA degradation.

    Science.gov (United States)

    Petit, F; Jarrousse, A S; Dahlmann, B; Sobek, A; Hendil, K B; Buri, J; Briand, Y; Schmid, H P

    1997-01-01

    We have identified two distinct subunits of 20 S proteasomes that are associated with RNase activity. Proteasome subunits zeta and iota, eluted from two-dimensional Western blots, hydrolysed tobacco mosaic virus RNA, whereas none of the other subunits degraded this substrate under the same conditions. Additionally, proteasomes were dissociated by 6 M urea, and subunit zeta, containing the highest RNase activity, was isolated by anion-exchange chromatography and gel filtration. Purified subunit zeta migrated as a single spot on two-dimensional PAGE with a molecular mass of approx. 28 kDa. Addition of anti-(subunit zeta) antibodies led to the co-precipitation of this proteasome subunit and nuclease activity. This is the first evidence that proteasomal alpha-type subunits are associated with an enzymic activity, and our results provide further evidence that proteasomes may be involved in cellular RNA metabolism. PMID:9337855

  4. Market and Policy Barriers for Demand Response Providing Ancillary Services in U.S. Markets

    Energy Technology Data Exchange (ETDEWEB)

    Cappers, Peter [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); MacDonald, Jason [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Goldman, Charles [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

    2013-03-01

    In this study, we attempt to provide a comprehensive examination of various market and policy barriers to demand response providing ancillary services in both ISO/RTO and non-ISO/RTO regions, especially at the program provider level. It is useful to classify barriers in order to create a holistic understanding and identify parties that could be responsible for their removal. This study develops a typology of barriers focusing on smaller customers that must rely on a program provider (i.e., electric investor owned utility or IOU, ARC) to create an aggregated DR resource in order to bring ancillary services to the balancing authority.ii The barriers were identified through examinations of regulatory structures, market environments, and product offerings; and discussions with industry stakeholders and regulators. In order to help illustrate the differences in barriers among various wholesale market designs and their constituent retail environments, four regions were chosen to use as case studies: Colorado, Texas, Wisconsin, and New Jersey. We highlight the experience in each area as it relates to the identified barriers.

  5. Access to Emissions Distributions and Related Ancillary Data through the ECCAD database

    Science.gov (United States)

    Darras, Sabine; Granier, Claire; Liousse, Catherine; De Graaf, Erica; Enriquez, Edgar; Boulanger, Damien; Brissebrat, Guillaume

    2017-04-01

    The ECCAD database (Emissions of atmospheric Compounds and Compilation of Ancillary Data) provides a user-friendly access to global and regional surface emissions for a large set of chemical compounds and ancillary data (land use, active fires, burned areas, population,etc). The emissions inventories are time series gridded data at spatial resolution from 1x1 to 0.1x0.1 degrees. ECCAD is the emissions database of the GEIA (Global Emissions InitiAtive) project and a sub-project of the French Atmospheric Data Center AERIS (http://www.aeris-data.fr). ECCAD has currently more than 2200 users originating from more than 80 countries. The project benefits from this large international community of users to expand the number of emission datasets made available. ECCAD provides detailed metadata for each of the datasets and various tools for data visualization, for computing global and regional totals and for interactive spatial and temporal analysis. The data can be downloaded as interoperable NetCDF CF-compliant files, i.e. the data are compatible with many other client interfaces. The presentation will provide information on the datasets available within ECCAD, as well as examples of the analysis work that can be done online through the website: http://eccad.aeris-data.fr.

  6. Safe introduction of ancillary trocars in gynecological surgery: the "yellow island" anatomical landmark.

    Science.gov (United States)

    Vitale, Salvatore Giovanni; Gasbarro, Nicola; Lagana, Antonio Simone; Sapia, Fabrizio; Rapisarda, A M C; Valenti, Gaetano; Trovato, Maria Antonietta; Rossetti, Diego; Chiofalo, Benito; Barrasso, Giuseppina; Tinelli, Andrea; Corrado, Francesco

    2016-01-01

    This technical note aims to suggest a safe introduction of ancillary trocars in gynaecological surgery, in order to prevent inferior epigastric artery damage. We performed a narrative overview, synthesizing the findings of literature retrieved from searches of computerized databases. Among the different techniques, the identification of the "yellow island" as anatomical landmark seems to be a useful aid to avoid complication. This particular landmark is identified taking the lateral third of a line between the anterior superior iliac spine and the umbilicus as reference points, by a subperitoneal accumulation of adipose tissue located in that area. "Yellow island" could be considered a safe place for trocars introduction because epigastric artery never runs there, even in patients with particular anatomical variants. This technique is particularly useful in obese patients, in which "yellow island" appears to be more evident respect to lean ones. Furthermore, the use of "open" trocars insertion may reduce the possibility of epigastric artery lesions respect to "Verres needle technique". The identification of the "yellow island" anatomical landmark could be considered an useful aid for the safe introduction of ancillary trocars in gynaecological surgery. Gynaecology, Laparoscopic surgery, Trocars placement.

  7. BLINK REFLEX IN MULTIPLE SCLEROSIS: AN ANCILLARY TEST FOR DETECTING BRAIN STEM LESIONS

    Directory of Open Access Journals (Sweden)

    M ETEMEDYFAR

    2001-09-01

    Full Text Available Introduction. Electrodiagnostic tests are one of the ancillary procedures that are used for diagnosis of multiple sclerosis (MS. This study investigates the frequency of abnormal blink reflex in patients with MS. Methods. In this cross sectional diagnostic study, 100 patients (26 male and 74 female with definite MS were selected based on clinical and MRI findings. they were referred to Al- zahra hospital (affiliated to iUMSHS during year 2000. Blink reflex (BR waves including R1, R2, R2 were recorded inpatients through the stimulation of supraorbital nerve. Results. The frequency of abnormal BR in MS patients with brain stem involvement was 77.9 percent and in those without brain stem involvement was 36.6 percent (P < 0.001. There was a significant relationship between the duration of MS and the abnormality in BR. Discussion. The frequency of abnormal blink reflex in MS is significantly associated with site of involvement in the brain. The majority of MS patients with brain stem involvement have abnormal BR. It is proposed that in patients with symptoms and signs of MS if there was no accessibility for MRI or if the results of MRI were equivocal, blink reflex test should be performed in addition to other ancillary tests.

  8. Urology Group Compensation and Ancillary Service Models in an Era of Value-based Care.

    Science.gov (United States)

    Shore, Neal D; Jacoby, Dana

    2016-01-01

    Changes involving the health care economic landscape have affected physicians' workflow, productivity, compensation structures, and culture. Ongoing Federal legislation regarding regulatory documentation and imminent payment-changing methodologies have encouraged physician consolidation into larger practices, creating affiliations with hospitals, multidisciplinary medical specialties, and integrated delivery networks. As subspecialization and evolution of care models have accelerated, independent medical groups have broadened ancillary service lines by investing in enterprises that compete with hospital-based (academic and nonacademic) entities, as well as non-physician- owned multispecialty enterprises, for both outpatient and inpatient services. The looming and dramatic shift from volume- to value-based health care compensation will assuredly affect urology group compensation arrangements and productivity formulae. For groups that can implement change rapidly, efficiently, and harmoniously, there will be opportunities to achieve the Triple Aim goals of the Patient Protection and Affordable Care Act, while maintaining a successful medical-financial practice. In summary, implementing new payment algorithms alongside comprehensive care coordination will assist urology groups in addressing the health economic cost and quality challenges that have been historically encountered with fee-for-service systems. Urology group leadership and stakeholders will need to adjust internal processes, methods of care coordination, cultural dependency, and organizational structures in order to create better systems of care and management. In response, ancillary services and patient throughput will need to evolve in order to adequately align quality measurement and reporting systems across provider footprints and patient populations.

  9. Open Automated Demand Response Communications in Demand Response for Wholesale Ancillary Services

    Energy Technology Data Exchange (ETDEWEB)

    Kiliccote, Sila; Piette, Mary Ann; Ghatikar, Girish; Koch, Ed; Hennage, Dan; Hernandez, John; Chiu, Albert; Sezgen, Osman; Goodin, John

    2009-11-06

    The Pacific Gas and Electric Company (PG&E) is conducting a pilot program to investigate the technical feasibility of bidding certain demand response (DR) resources into the California Independent System Operator's (CAISO) day-ahead market for ancillary services nonspinning reserve. Three facilities, a retail store, a local government office building, and a bakery, are recruited into the pilot program. For each facility, hourly demand, and load curtailment potential are forecasted two days ahead and submitted to the CAISO the day before the operation as an available resource. These DR resources are optimized against all other generation resources in the CAISO ancillary service. Each facility is equipped with four-second real time telemetry equipment to ensure resource accountability and visibility to CAISO operators. When CAISO requests DR resources, PG&E's OpenADR (Open Automated DR) communications infrastructure is utilized to deliver DR signals to the facilities energy management and control systems (EMCS). The pre-programmed DR strategies are triggered without a human in the loop. This paper describes the automated system architecture and the flow of information to trigger and monitor the performance of the DR events. We outline the DR strategies at each of the participating facilities. At one site a real time electric measurement feedback loop is implemented to assure the delivery of CAISO dispatched demand reductions. Finally, we present results from each of the facilities and discuss findings.

  10. allelic variation of hmw glutenin subunits of ethiopian bread wheat ...

    African Journals Online (AJOL)

    journal

    reduced subunits of glutenin proteins bands are separated: the high molecular weight (HMW) and low molecular weight (LMW) subunits (Payne et al.,1980; Jackson et al., 1983). The HMW glutenin subunits (GS) of wheat protein are quantitatively minor, but functionally an important group of gluten proteins in the process of ...

  11. DNA sequences, recombinant DNA molecules and processes for producing the A and B subunits of cholera toxin and preparations containing so-obtained subunit or subunits

    Energy Technology Data Exchange (ETDEWEB)

    Harford, N.; De Wilde, M.

    1987-05-19

    A recombinant DNA molecule is described comprising at least a portion coding for subunits A and B of cholera toxin, or a fragment or derivative of the portion wherein the fragment or derivative codes for a polypeptide have an activity which can induce an immune response to subunit A; can induce an immune response to subunit A and cause epithelial cell penetration and the enzymatic effect leading to net loss of fluid into the gut lumen; can bind to the membrane receptor for the B subunit of cholera toxin; can induce an immune response to subunit B; can induce an immune response to subunit B and bind to the membrane receptor; or has a combination of the activities.

  12. An assessment of market and policy barriers for demand response providing ancillary services in U.S. electricity markets

    International Nuclear Information System (INIS)

    Cappers, Peter; MacDonald, Jason; Goldman, Charles; Ma, Ookie

    2013-01-01

    An impact of increased variable renewable generation is the need for balancing authorities to procure more ancillary services. While demand response resources are technically capable of providing these services, current experience across the U.S. illustrates they are relatively minor players in most regions. Accessing demand response resources for ancillary services may require a number of changes to policies and common practices at multiple levels. Regional reliability councils must first define ancillary services such that demand response resources may provide them. Once the opportunity exists, balancing authorities define and promulgate rules that set the infrastructure investments and performance attributes of a resource wishing to provide such services. These rules also dictate expected revenue streams which reveal the cost effectiveness of these resources. The regulatory compact between utility and state regulators, along with other statutes and decisions by state policymakers, may impact the interest of demand response program providers to pursue these resources as ancillary service providers. This paper identifies within these broad categories specific market and policy barriers to demand response providing ancillary services in different wholesale and retail environments, with emphasis on smaller customers who must be aggregated through a program provider to meet minimum size requirements for wholesale transactions. - Highlights: • We identify barriers keeping demand response from providing ancillary services. • Institutional, financial and program provider business model barriers exist. • Product definitions and rules do not always accommodate demand response well. • Expected revenues are uncertain and may not exceed required investments costs. • Regulatory compact and state statutes limit opportunities for program providers

  13. Staff working in ancillary departments at a tertiary care hospital in Bengaluru, Karnataka, India: How healthy are they?

    Science.gov (United States)

    Balasubramanya, Bhavya; Nisha, Catherin; Ramesh, Naveen; Joseph, Bobby

    2016-01-01

    Background: Ancillary health services are those supplemental services other than room, board, and medical/nursing services provided to hospital patients in the course of care. Ancillary department staff forms an integral part in the smooth functioning of a hospital. There is a need to focus on the health of these individuals to ensure their well-being and in turn, productivity at the workplace. Objective: To study the morbidity profile of the staff working at ancillary departments of a tertiary care hospital in Bengaluru, Karnataka, India. Materials and Methods: We conducted our study in a 1,200-bedded tertiary care hospital in Bengaluru, Karnataka, India. Annual medical checkup (AMC) for all the staff working at the ancillary departments has been started in recent years and is provided free of cost and during working hours. A total of 150 employees from ancillary departments underwent AMC in the year 2013. Data were analyzed using Statistical Package for the Social Sciences (SPSS) version 20.0. Spearman's correlation and Chi-square test were used. Results: Of the 150 employees, the majority was male (72%); the mean age was 38 ± 11 years. The most common morbidities were diabetes mellitus (11%), hypertension (10.6%), musculoskeletal disorders (9.3%), surgical problems (8.6%, hemorrhoids, varicose veins), and dental caries (6.6%). On stool microscopy, 12% of the dietary workers showed ova/cyst. There was a significant positive correlation between age and the number of chronic morbidities (P < 0.01). Conclusion: Lifestyle disorders such as diabetes mellitus and hypertension were the major morbidities among the staff in the ancillary departments of the hospital. We ensured regular follow-up, adherence to medication, and lifestyle modifications in terms of diet and exercise. PMID:27390479

  14. Ground State and Excited State Tuning in Ferric Dipyrrin Complexes Promoted by Ancillary Ligand Exchange

    Energy Technology Data Exchange (ETDEWEB)

    Kleinlein, Claudia; Zheng, Shao-Liang; Betley, Theodore A.

    2017-04-24

    Three ferric dipyrromethene complexes featuring different ancillary ligands were synthesized by one electron oxidation of ferrous precursors. Four-coordinate iron complexes of the type (ArL)FeX2 [ArL = 1,9-(2,4,6-Ph3C6H2)2-5-mesityldipyrromethene] with X = Cl or tBuO were prepared and found to be high-spin (S = 5/2), as determined by superconducting quantum interference device magnetometry, electron paramagnetic resonance, and 57Fe Mössbauer spectroscopy. The ancillary ligand substitution was found to affect both ground state and excited properties of the ferric complexes examined. While each ferric complex displays reversible reduction and oxidation events, each alkoxide for chloride substitution results in a nearly 600 mV cathodic shift of the FeIII/II couple. The oxidation event remains largely unaffected by the ancillary ligand substitution and is likely dipyrrin-centered. While the alkoxide substituted ferric species largely retain the color of their ferrous precursors, characteristic of dipyrrin-based ligand-to-ligand charge transfer (LLCT), the dichloride ferric complex loses the prominent dipyrrin chromophore, taking on a deep green color. Time-dependent density functional theory analyses indicate the weaker-field chloride ligands allow substantial configuration mixing of ligand-to-metal charge transfer into the LLCT bands, giving rise to the color changes observed. Furthermore, the higher degree of covalency between the alkoxide ferric centers is manifest in the observed reactivity. Delocalization of spin density onto the tert-butoxide ligand in (ArL)FeCl(OtBu) is evidenced by hydrogen atom abstraction to yield (ArL)FeCl and HOtBu in the presence of substrates containing weak C–H bonds, whereas the chloride (ArL)FeCl2 analogue does not react under these conditions.

  15. Utilization of Ancillary Data Sets for Conceptual SMAP Mission Algorithm Development and Product Generation

    Science.gov (United States)

    O'Neill, P.; Podest, E.

    2011-01-01

    The planned Soil Moisture Active Passive (SMAP) mission is one of the first Earth observation satellites being developed by NASA in response to the National Research Council's Decadal Survey, Earth Science and Applications from Space: National Imperatives for the Next Decade and Beyond [1]. Scheduled to launch late in 2014, the proposed SMAP mission would provide high resolution and frequent revisit global mapping of soil moisture and freeze/thaw state, utilizing enhanced Radio Frequency Interference (RFI) mitigation approaches to collect new measurements of the hydrological condition of the Earth's surface. The SMAP instrument design incorporates an L-band radar (3 km) and an L band radiometer (40 km) sharing a single 6-meter rotating mesh antenna to provide measurements of soil moisture and landscape freeze/thaw state [2]. These observations would (1) improve our understanding of linkages between the Earth's water, energy, and carbon cycles, (2) benefit many application areas including numerical weather and climate prediction, flood and drought monitoring, agricultural productivity, human health, and national security, (3) help to address priority questions on climate change, and (4) potentially provide continuity with brightness temperature and soil moisture measurements from ESA's SMOS (Soil Moisture Ocean Salinity) and NASA's Aquarius missions. In the planned SMAP mission prelaunch time frame, baseline algorithms are being developed for generating (1) soil moisture products both from radiometer measurements on a 36 km grid and from combined radar/radiometer measurements on a 9 km grid, and (2) freeze/thaw products from radar measurements on a 3 km grid. These retrieval algorithms need a variety of global ancillary data, both static and dynamic, to run the retrieval models, constrain the retrievals, and provide flags for indicating retrieval quality. The choice of which ancillary dataset to use for a particular SMAP product would be based on a number of factors

  16. Liposome-Based Adjuvants for Subunit Vaccines: Formulation Strategies for Subunit Antigens and Immunostimulators

    DEFF Research Database (Denmark)

    Schmidt, Signe Tandrup; Foged, Camilla; Korsholm, Karen Smith

    2016-01-01

    for which no effective vaccines exist. The subunit vaccine technology exploits pathogen subunits as antigens, e.g., recombinant proteins or synthetic peptides, allowing for highly specific immune responses against the pathogens. However, such antigens are usually not sufficiently immunogenic to induce......The development of subunit vaccines has become very attractive in recent years due to their superior safety profiles as compared to traditional vaccines based on live attenuated or whole inactivated pathogens, and there is an unmet medical need for improved vaccines and vaccines against pathogens...... been licensed for use in human vaccines, and they mainly stimulate humoral immunity. Thus, there is an unmet demand for the development of safe and efficient adjuvant systems that can also stimulate cell-mediated immunity (CMI). Adjuvants constitute a heterogeneous group of compounds, which can broadly...

  17. Ancillary testing, diagnostic/classification criteria and severity grading in Behçet disease.

    Science.gov (United States)

    Okada, Annabelle A; Stanford, Miles; Tabbara, Khalid

    2012-12-01

    Since there is no pathognomonic clinical sign or laboratory test to distinguish Behçet disease from other uveitic entities, the diagnosis must be made based on characteristic ocular and systemic findings in the absence of evidence of other disease that can explain the findings. Ancillary tests, including ocular and brain imaging studies, are used to assess the severity of intraocular inflammation and systemic manifestations of Behçet disease, to identify latent infections and other medical conditions that might worsen with systemic treatment, and to monitor for adverse effects of drugs used. There are two diagnostic or classification criteria in general use by the uveitis community, one from Japan and one from an international group; both rely on a minimum number and/or combination of clinical findings to identify Behçet disease. Finally, several grading schemes have been proposed to assess severity of ocular disease and response to treatment.

  18. Ancillary Services for the European Grid with High Shares of Wind and Solar Power

    DEFF Research Database (Denmark)

    Van Hulle, Frans; Holttinen, Hannele; Kiviluoma, Juha

    2012-01-01

    services that are especially relevant for wind and solar power. These are mainly related to frequency, voltage control and restoration of the system. In addition, based on existing experience and wind integration studies, the paper analyses impacts that high amounts of wind/solar will have on different......With significantly increasing share of variable renewable power generation like wind and solar PV, the need in the power system for ancillary services supporting the network frequency, voltage, etc. changes. Turning this issue around, market opportunities will emerge for wind and solar PV...... technology to deliver such grid services. In the European power system, adequate market mechanisms need to be developed to ensure that there will be an efficient trading of these services. For that purpose a range of (economic) characteristics of wind (and solar) power as providers of grid services need...

  19. Critical Joints in Large Composite Primary Aircraft Structures. Volume 3: Ancillary Test Results

    Science.gov (United States)

    Bunin, Bruce L.; Sagui, R. L.

    1985-01-01

    A program was conducted to develop the technology for critical structural joints for composite wing structure that meets all the design requirements of a 1990 commercial transport aircraft. The results of a comprehensive ancillary test program are summarized, consisting of single-bolt composite joint specimens tested in a variety of configurations. These tests were conducted to characterize the strength and load deflection properties that are required for multirow joint analysis. The composite material was Toray 300 fiber and Ciba-Geigy 914 resin, in the form of 0.005 and 0.01 inch thick unidirectional tape. Tests were conducted in single and double shear for loaded and unloaded hole configurations under both tensile and compressive loading. Two different layup patterns were examined. All tests were conducted at room temperature. In addition, the results of NASA Standard Toughness Test (NASA RP 1092) are reported, which were conducted for several material systems.

  20. Engineering Evaluation/Cost Analysis for the 100-N Area Ancillary Facilities and Integration Plan

    International Nuclear Information System (INIS)

    Mukherjee, B.

    1997-09-01

    This document presents the results of an engineering evaluation/cost analysis (EE/CA) that was conducted to evaluate alternatives for addressing final disposition of contaminated buildings and structures in the 100-N Area of the Hanford Site. The Hanford Site is located in southeastern Washington State and is owned by the U.S. Government and operated by the U.S. Department of Energy, Richland Operations Office (RL). In November 1989, the 100 Area of the Hanford Site (as well as the 200, 300, and 1100 Areas) was placed on the U.S. Environmental Protection Agency's National Priorities List (NPL) under the Comprehensive Environmental Response, Compensation, and Liability Act of 1980. The 100 Area NPL includes the 100-N Area, which is in various stages of the remediation process. It has been determined by RL that hazardous substances in the 100-N Area ancillary facilities may present a potential threat to human health or the environment, and that a non-time critical removal action at these facilities is warranted. To help determine the most appropriate action, RL, in cooperation with the Washington State Department of Ecology (Ecology) and the EPA, has prepared this EE/CA. The scope of the evaluation includes the inactive contaminated ancillary facilities in the 100-N Area, the facilities residing in the buffer zone, and the Hanford Generating Plant (HGP) and the solid waste management units (SWMUs) inside HGP support facilities. The 105-N Reactor and 109-N Heat Exchange facilities are excluded from this EE/CA evaluation

  1. Development of modified rational buyer auction for procurement of ancillary services utilizing participation matrix

    International Nuclear Information System (INIS)

    Jamalzadeh, R.; Ardehali, M.M.; Rashidinejad, M.

    2008-01-01

    The rational buyer auction is based on a simultaneous auction that is the redesigned auction mechanism for ancillary services by the California independent system operator (CAISO). The incentive for the rational buyer auction is CAISO's intent to adopt a common sense rule of substituting higher-quality lower-cost services for lower-quality higher-cost services, when it results in reduced total procurement cost. For the purposes of designing a desirable auction where the minimum cost for the objective function as well as prevention of price reversal are achieved, either the payment cost in marginal pricing auction must be lowered or price reversal in rational buyer auction must be avoided. The objectives of this study are (a) to develop a newly proposed 'modified rational buyer' auction that does not allow price reversal and (b) to propose and validate a solution procedure that is based on participation matrix and discrete programming. The validation of the proposed solution procedure is accomplished through examination of two case studies available in the literature. Based on the first case study data, it is shown that the newly developed modified rational buyer auction avoids price reversal occurrence, while the total payment of ISO is increased by 5.8%, as compared with rational buyer auction. Also in comparison with marginal pricing auction, the ISO payment is lowered by 38.8%, when the newly developed modified rational buyer auction is utilized. For future work, it is recommended that the application of the modified rational buyer auction to joint dispatch of energy and ancillary services is investigated. (author)

  2. MspA nanopores from subunit dimers.

    Directory of Open Access Journals (Sweden)

    Mikhail Pavlenok

    Full Text Available Mycobacterium smegmatis porin A (MspA forms an octameric channel and represents the founding member of a new family of pore proteins. Control of subunit stoichiometry is important to tailor MspA for nanotechnological applications. In this study, two MspA monomers were connected by linkers ranging from 17 to 62 amino acids in length. The oligomeric pore proteins were purified from M. smegmatis and were shown to form functional channels in lipid bilayer experiments. These results indicated that the peptide linkers did not prohibit correct folding and localization of MspA. However, expression levels were reduced by 10-fold compared to wild-type MspA. MspA is ideal for nanopore sequencing due to its unique pore geometry and its robustness. To assess the usefulness of MspA made from dimeric subunits for DNA sequencing, we linked two M1-MspA monomers, whose constriction zones were modified to enable DNA translocation. Lipid bilayer experiments demonstrated that this construct also formed functional channels. Voltage gating of MspA pores made from M1 monomers and M1-M1 dimers was identical indicating similar structural and dynamic channel properties. Glucose uptake in M. smegmatis cells lacking porins was restored by expressing the dimeric mspA M1 gene indicating correct folding and localization of M1-M1 pores in their native membrane. Single-stranded DNA hairpins produced identical ionic current blockades in pores made from monomers and subunit dimers demonstrating that M1-M1 pores are suitable for DNA sequencing. This study provides the proof of principle that production of single-chain MspA pores in M. smegmatis is feasible and paves the way for generating MspA pores with altered stoichiometries. Subunit dimers enable better control of the chemical and physical properties of the constriction zone of MspA. This approach will be valuable both in understanding transport across the outer membrane in mycobacteria and in tailoring MspA for nanopore

  3. Glycine receptor subunits expression in the developing rat retina.

    Science.gov (United States)

    Sánchez-Chávez, Gustavo; Velázquez-Flores, Miguel Ángel; Ruiz Esparza-Garrido, Ruth; Salceda, Rocío

    2017-09-01

    Glycine receptor (GlyR) consists of two α (1-4) and three β subunits. Considerable evidence indicates that the adult retina expresses the four types of α subunits; however, the proportion of these subunits in adult and immature retina is almost unknown. In this report we have studied mRNA and the protein expression of GlyR subunits in the retina during postnatal rat development by Real-Time qRT-PCR and western blot. mRNA and protein expression indicated a gradual increase of the α1, α3, α4 and β GlyR subunits during postnatal ages tested. The mRNA β subunit showed higher expression levels (∼3 fold) than those observed for the α1 and α3 subunits. Very interestingly, the α2 GlyR subunit had the highest expression in the retina, even in the adult. These results revealed the expression of GlyR at early postnatal ages, supporting its role in retina development. In addition, our results indicated that the adult retina expressed a high proportion of the α2 subunit, suggesting the expression of monomeric and/or heteromeric receptors. A variety of studies are needed to further characterize the role of the specific subunits in both adult and immature retina. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. 76 FR 36400 - Third-Party Provision of Ancillary Services; Accounting and Financial Reporting for New Electric...

    Science.gov (United States)

    2011-06-22

    ...-000 and AD10-13-000] Third-Party Provision of Ancillary Services; Accounting and Financial Reporting... Commission's accounting and financial reporting requirements currently do not contain specific accounting \\12... financial reporting requirements \\41\\ for public utilities \\42\\ are designed to provide information about a...

  5. 47 CFR 25.252 - Special requirements for ancillary terrestrial components operating in the 2000-2020 MHz/2180...

    Science.gov (United States)

    2010-10-01

    ... components operating in the 2000-2020 MHz/2180-2200 MHz bands. 25.252 Section 25.252 Telecommunication... Standards § 25.252 Special requirements for ancillary terrestrial components operating in the 2000-2020 MHz... equivalent. (c) For ATC operations in the 2000-2020 MHz band, the power of any emission outside the licensee...

  6. 75 FR 66110 - Guidelines for Use of Stored Specimens and Access to Ancillary Data and Proposed Cost Schedule...

    Science.gov (United States)

    2010-10-27

    ... scientific papers with notable findings. Procedures for Proposals: All investigators (including NIH and NICHD... Proposal: Proposals can be submitted on an ongoing basis. Scientific Review Dates: Technical Panels for... ancillary data will be evaluated by an ad hoc Technical Panel for scientific merit. The BRADSC will...

  7. 41 CFR 102-72.67 - What work is covered under an ancillary repair and alteration delegation?

    Science.gov (United States)

    2010-07-01

    ... 41 Public Contracts and Property Management 3 2010-07-01 2010-07-01 false What work is covered under an ancillary repair and alteration delegation? 102-72.67 Section 102-72.67 Public Contracts and Property Management Federal Property Management Regulations System (Continued) FEDERAL MANAGEMENT REGULATION REAL PROPERTY 72-DELEGATION OF AUTHORITY...

  8. Subunit Arrangement and Function in NMDA Receptors

    Energy Technology Data Exchange (ETDEWEB)

    Furukawa,H.; Singh, S.; Mancusso, R.; Gouaux, E.

    2005-01-01

    Excitatory neurotransmission mediated by NMDA (N-methyl-D-aspartate) receptors is fundamental to the physiology of the mammalian central nervous system. These receptors are heteromeric ion channels that for activation require binding of glycine and glutamate to the NR1 and NR2 subunits, respectively. NMDA receptor function is characterized by slow channel opening and deactivation, and the resulting influx of cations initiates signal transduction cascades that are crucial to higher functions including learning and memory. Here we report crystal structures of the ligand-binding core of NR2A with glutamate and that of the NR1-NR2A heterodimer with glutamate and glycine. The NR2A-glutamate complex defines the determinants of glutamate and NMDA recognition, and the NR1-NR2A heterodimer suggests a mechanism for ligand-induced ion channel opening. Analysis of the heterodimer interface, together with biochemical and electrophysiological experiments, confirms that the NR1-NR2A heterodimer is the functional unit in tetrameric NMDA receptors and that tyrosine 535 of NR1, located in the subunit interface, modulates the rate of ion channel deactivation.

  9. Sodium channel β subunits: emerging targets in channelopathies.

    Science.gov (United States)

    O'Malley, Heather A; Isom, Lori L

    2015-01-01

    Voltage-gated sodium channels (VGSCs) are responsible for the initiation and propagation of action potentials in excitable cells. VGSCs in mammalian brain are heterotrimeric complexes of α and β subunits. Although β subunits were originally termed auxiliary, we now know that they are multifunctional signaling molecules that play roles in both excitable and nonexcitable cell types and with or without the pore-forming α subunit present. β subunits function in VGSC and potassium channel modulation, cell adhesion, and gene regulation, with particularly important roles in brain development. Mutations in the genes encoding β subunits are linked to a number of diseases, including epilepsy, sudden death syndromes like SUDEP and SIDS, and cardiac arrhythmia. Although VGSC β subunit-specific drugs have not yet been developed, this protein family is an emerging therapeutic target.

  10. Transcriptional regulators of Na, K-ATPase subunits

    OpenAIRE

    Zhiqin eLi; Sigrid A Langhans

    2015-01-01

    The Na,K-ATPase classically serves as an ion pump creating an electrochemical gradient across the plasma membrane that is essential for transepithelial transport, nutrient uptake and membrane potential. In addition, Na,K-ATPase also functions as a receptor, a signal transducer and a cell adhesion molecule. With such diverse roles, it is understandable that the Na,K-ATPase subunits, the catalytic alpha-subunit, the beta-subunit and the FXYD proteins, are controlled extensively during developme...

  11. Preliminary Results of Ancillary Safety Analyses Supporting TREAT LEU Conversion Activities

    Energy Technology Data Exchange (ETDEWEB)

    Brunett, A. J. [Argonne National Lab. (ANL), Argonne, IL (United States); Fei, T. [Argonne National Lab. (ANL), Argonne, IL (United States); Strons, P. S. [Argonne National Lab. (ANL), Argonne, IL (United States); Papadias, D. D. [Argonne National Lab. (ANL), Argonne, IL (United States); Hoffman, E. A. [Argonne National Lab. (ANL), Argonne, IL (United States); Kontogeorgakos, D. C. [Argonne National Lab. (ANL), Argonne, IL (United States); Connaway, H. M. [Argonne National Lab. (ANL), Argonne, IL (United States); Wright, A. E. [Argonne National Lab. (ANL), Argonne, IL (United States)

    2015-10-01

    The Transient Reactor Test Facility (TREAT), located at Idaho National Laboratory (INL), is a test facility designed to evaluate the performance of reactor fuels and materials under transient accident conditions. The facility, an air-cooled, graphite-moderated reactor designed to utilize fuel containing high-enriched uranium (HEU), has been in non-operational standby status since 1994. Currently, in support of the missions of the Department of Energy (DOE) National Nuclear Security Administration (NNSA) Material Management and Minimization (M3) Reactor Conversion Program, a new core design is being developed for TREAT that will utilize low-enriched uranium (LEU). The primary objective of this conversion effort is to design an LEU core that is capable of meeting the performance characteristics of the existing HEU core. Minimal, if any, changes are anticipated for the supporting systems (e.g. reactor trip system, filtration/cooling system, etc.); therefore, the LEU core must also be able to function with the existing supporting systems, and must also satisfy acceptable safety limits. In support of the LEU conversion effort, a range of ancillary safety analyses are required to evaluate the LEU core operation relative to that of the existing facility. These analyses cover neutronics, shielding, and thermal hydraulic topics that have been identified as having the potential to have reduced safety margins due to conversion to LEU fuel, or are required to support the required safety analyses documentation. The majority of these ancillary tasks have been identified in [1] and [2]. The purpose of this report is to document the ancillary safety analyses that have been performed at Argonne National Laboratory during the early stages of the LEU design effort, and to describe ongoing and anticipated analyses. For all analyses presented in this report, methodologies are utilized that are consistent with, or improved from, those used in analyses for the HEU Final Safety Analysis

  12. Approaches to Enable Demand Response by Industrial Loads for Ancillary Services Provision

    Science.gov (United States)

    Zhang, Xiao

    Demand response has gained significant attention in recent years as it demonstrates potentials to enhance the power system's operational flexibility in a cost-effective way. Industrial loads such as aluminum smelters, steel manufacturers, and cement plants demonstrate advantages in supporting power system operation through demand response programs, because of their intensive power consumption, already existing advanced monitoring and control infrastructure, and the strong economic incentive due to the high energy costs. In this thesis, we study approaches to efficiently integrate each of these types of manufacturing processes as demand response resources. The aluminum smelting process is able to change its power consumption both accurately and quickly by controlling the pots' DC voltage, without affecting the production quality. Hence, an aluminum smelter has both the motivation and the ability to participate in demand response. First, we focus on determining the optimal regulation capacity that such a manufacturing plant should provide. Next, we focus on determining its optimal bidding strategy in the day-ahead energy and ancillary services markets. Electric arc furnaces (EAFs) in steel manufacturing consume a large amount of electric energy. However, a steel plant can take advantage of time-based electricity prices by optimally arranging energy-consuming activities to avoid peak hours. We first propose scheduling methods that incorporate the EAFs' flexibilities to reduce the electricity cost. We then propose methods to make the computations more tractable. Finally, we extend the scheduling formulations to enable the provision of spinning reserve. Cement plants are able to quickly adjust their power consumption rate by switching on/off the crushers. However, switching on/off the loading units only achieves discrete power changes, which restricts the load from offering valuable ancillary services such as regulation and load following, as continuous power changes

  13. Using remote sensing and ancillary data to extend airborne electromagnetic resistivity surveys for regional permafrost interpretation

    Science.gov (United States)

    Pastick, N.; Wylie, B. K.; Minsley, B. J.; Jorgenson, T. T.; Ji, L.; Walvoord, M. A.; Smith, B. D.; Abraham, J. D.; Rose, J.

    2011-12-01

    Permafrost has a significant impact on high latitude ecosystems and is spatially heterogeneous. However, only generalized maps of permafrost extent are available. Due to its impacts on subsurface hydrology, lake water levels, vegetation communities, and surface soil deformations, understanding the spatial extents and depth of permafrost are critical. Electrical resistivity increases dramatically as a soil freezes and can be used as a proxy for permafrost presence particularly if the underlying soils and geologic characteristics are understood. An airborne electromagnetic survey (AEM) was conducted over a portion of the Yukon Flats ecoregion in central Alaska with measurements taken in both reconnaissance lines and contiguous block area coverage. The AEM was flown in June 2010 and subsurface resistivity models were derived by inverting the AEM data. Landsat TM at-sensor reflectance, thermal, and spectral index data from late August to early September 2008, Digital Elevation Models (DEM) and derivatives, and other ancillary data were used in a regression tree model to predict near surface electrical resistivity at the 0-1m and the 0-2.6m depth intervals. AEM locations from homogenous landsat 90 m by 90 m windows were randomly separated into a training set for model development (n = 8,848) and an impendent test data set (n = 988) for model accuracy assessment. Model development and independent test accuracies for 0-1 m electric resistivity had training and test R2 values of 0.90 and 0.87, respectively, and for the 0-2.6m electric resistivity training and test R2 values were also 0.90 and 0.87, respectively, which indicated accurate prediction models. Important variables for stratifying the various piecewise regressions were elevation and averaged 2000-2008 ecosystem performance anomalies. Important independent variables used in the multiple regression equations were the Normalized Difference Infrared Index (NDII), NDII7 (NDII using band 7), soil moisture mapped from

  14. The SPICE concept - An approach to providing geometric and other ancillary information needed for interpretation of data returned from space science instruments

    Science.gov (United States)

    Acton, Charles H., Jr.

    1990-01-01

    The Navigation Ancillary Information Facility (NAIF), acting under the direction of NASA's Office of Space Science and Applications, and with substantial participation of the planetary science community, is designing and implementing an ancillary data system - called SPICE - to assist scientists in planning and interpreting scientific observations taken from spaceborne instruments. The principal objective of the implemented SPICE system is that it will hold the essential geometric and related ancillary information needed to recover the full value of science instrument data, and that it will facilitate correlations of individual instrument datasets with data obtained from other instruments on the same or other spacecraft.

  15. Liposome-Based Adjuvants for Subunit Vaccines: Formulation Strategies for Subunit Antigens and Immunostimulators

    Directory of Open Access Journals (Sweden)

    Signe Tandrup Schmidt

    2016-03-01

    Full Text Available The development of subunit vaccines has become very attractive in recent years due to their superior safety profiles as compared to traditional vaccines based on live attenuated or whole inactivated pathogens, and there is an unmet medical need for improved vaccines and vaccines against pathogens for which no effective vaccines exist. The subunit vaccine technology exploits pathogen subunits as antigens, e.g., recombinant proteins or synthetic peptides, allowing for highly specific immune responses against the pathogens. However, such antigens are usually not sufficiently immunogenic to induce protective immunity, and they are often combined with adjuvants to ensure robust immune responses. Adjuvants are capable of enhancing and/or modulating immune responses by exposing antigens to antigen-presenting cells (APCs concomitantly with conferring immune activation signals. Few adjuvant systems have been licensed for use in human vaccines, and they mainly stimulate humoral immunity. Thus, there is an unmet demand for the development of safe and efficient adjuvant systems that can also stimulate cell-mediated immunity (CMI. Adjuvants constitute a heterogeneous group of compounds, which can broadly be classified into delivery systems or immunostimulators. Liposomes are versatile delivery systems for antigens, and they can carefully be customized towards desired immune profiles by combining them with immunostimulators and optimizing their composition, physicochemical properties and antigen-loading mode. Immunostimulators represent highly diverse classes of molecules, e.g., lipids, nucleic acids, proteins and peptides, and they are ligands for pattern-recognition receptors (PRRs, which are differentially expressed on APC subsets. Different formulation strategies might thus be required for incorporation of immunostimulators and antigens, respectively, into liposomes, and the choice of immunostimulator should ideally be based on knowledge regarding the

  16. Disaggregating census data for population mapping using random forests with remotely-sensed and ancillary data.

    Science.gov (United States)

    Stevens, Forrest R; Gaughan, Andrea E; Linard, Catherine; Tatem, Andrew J

    2015-01-01

    High resolution, contemporary data on human population distributions are vital for measuring impacts of population growth, monitoring human-environment interactions and for planning and policy development. Many methods are used to disaggregate census data and predict population densities for finer scale, gridded population data sets. We present a new semi-automated dasymetric modeling approach that incorporates detailed census and ancillary data in a flexible, "Random Forest" estimation technique. We outline the combination of widely available, remotely-sensed and geospatial data that contribute to the modeled dasymetric weights and then use the Random Forest model to generate a gridded prediction of population density at ~100 m spatial resolution. This prediction layer is then used as the weighting surface to perform dasymetric redistribution of the census counts at a country level. As a case study we compare the new algorithm and its products for three countries (Vietnam, Cambodia, and Kenya) with other common gridded population data production methodologies. We discuss the advantages of the new method and increases over the accuracy and flexibility of those previous approaches. Finally, we outline how this algorithm will be extended to provide freely-available gridded population data sets for Africa, Asia and Latin America.

  17. Workshop on environmental assessment. [Regulation of applications of nuclear energy and related ancillary systems

    Energy Technology Data Exchange (ETDEWEB)

    Watson, E.C. (comp.)

    1982-07-01

    Objectives of the workshop were: to review and evaluate the state-of-the-art of environmental impact assessments as applied to the regulation of applications of nuclear energy and related ancillary systems; to identify areas where existing technology allows establishing acceptable methods or standard practices that will meet the requirements of the NRC regulations, standards and guides for both normal operations and off-standard conditions including accident considerations; to illuminate topics where existing models or analytical methods are deficient because of unverified assumptions, a paucity of empirical data, conflicting results reported in the literature or a need for observation of operation systems; to compile, analyze and synthesize a prioritized set of research needs to advance the state-of-the-art to the level which will meet all of the requirements of the Commission's regulations, standards and guides; and to develop bases for maintaining the core of regulatory guidance at the optimum level balancing technical capabilities with practical considerations of cost and value to the regulatory process. The discussion held in small group sessions on aquatic, atmospheric, and terrestrial pathways are presented. The following research needs were identified as common to all three groups: validation of models; characterization of source terms; development of screening techniques; basis for de minimis levels of contamination; and updating of objectives for environmental monitoring programs.

  18. Cardiovascular Fat, Menopause, and Sex Hormones in Women: The SWAN Cardiovascular Fat Ancillary Study.

    Science.gov (United States)

    El Khoudary, Samar R; Shields, Kelly J; Janssen, Imke; Hanley, Carrie; Budoff, Matthew J; Barinas-Mitchell, Emma; Everson-Rose, Susan A; Powell, Lynda H; Matthews, Karen A

    2015-09-01

    Cardiovascular risk increases in women after menopause. Mounting evidence demonstrates a role of cardiovascular fat (CF) in the pathogenesis of coronary heart disease, but no research has examined CF in relation to sex hormones or menopausal status in women. The objective was to determine the relationship between CF depots, menopausal status, and endogenous sex hormones. Cross-sectional and longitudinal study designs were used. The setting included the Study of Women's Health Across the Nation (SWAN) Heart and Cardiovascular Fat Ancillary Study. A total of 456 women (mean age, 50.75 y); 62% premenopausal/early perimenopausal, and 38% late peri-/postmenopausal. Menopausal status, endogenous sex hormones measured simultaneously with CF volumes, and circulating estradiol available 4.80 years (median) before CF measures. Volumes of CF (epicardial adipose tissue [EAT], paracardial adipose tissue [PAT], total heart adipose tissue [TAT = EAT + PAT], and aortic perivascular adipose tissue [PVAT]). In final models, late peri-/postmenopausal women had 9.88% more EAT, 20.72% more PAT, and 11.69% more TAT volumes than pre-/early perimenopausal women (P menopausal status. In final models, lower estradiol concentrations were associated with greater volumes of PAT and TAT (P hormones are associated with CF. Perhaps CF plays a role in the higher risk of coronary heart disease reported in women after menopause.

  19. Planck intermediate results XXIII. Galactic plane emission components derived from Planck with ancillary data

    DEFF Research Database (Denmark)

    Ade, P. A. R.; Aghanim, N.; Arnaud, M.

    2015-01-01

    and to clarify the relationship between the various components. The region of the Galactic plane covered is l = 300 ! 0 ! 60where star-formation is highest and the emission is strong enough to make meaningful component separation. The latitude widths in this longituderange lie between 1 and 2, which correspond......Planck data when combined with ancillary data provide a unique opportunity to separate the diuse emission components of the inner Galaxy.The purpose of the paper is to elucidate the morphology of the various emission components in the strong star-formation region lying inside thesolar radius...... along the Galactic plane using the wide frequency coverage of Planck(28.4–857 GHz) in combination with low-frequency radio data at 0.408–2.3 GHz plus WMAP data at 23–94 GHz, along with far-infrared (FIR) datafrom COBE-DIRBE and IRAS. The free-free component is determined from radio recombination line...

  20. Current perspectives on the use of ancillary materials for the manufacture of cellular therapies.

    Science.gov (United States)

    Solomon, Jennifer; Csontos, Lynn; Clarke, Dominic; Bonyhadi, Mark; Zylberberg, Claudia; McNiece, Ian; Kurtzberg, Joanne; Bell, Rosemarie; Deans, Robert

    2016-01-01

    Continued growth in the cell therapy industry and commercialization of cell therapies that successfully advance through clinical trials has led to increased awareness around the need for specialized and complex materials utilized in their manufacture. Ancillary materials (AMs) are components or reagents used during the manufacture of cell therapy products but are not intended to be part of the final products. Commonly, there are limitations in the availability of clinical-grade reagents used as AMs. Furthermore, AMs may affect the efficacy of the cell product and subsequent safety of the cell therapy for the patient. As such, AMs must be carefully selected and appropriately qualified during the cell therapy development process. However, the ongoing evolution of cell therapy research, limited number of clinical trials and registered cell therapy products results in the current absence of specific regulations governing the composition, compliance, and qualification of AMs often leads to confusion by suppliers and users in this field. Here we provide an overview and interpretation of the existing global framework surrounding AM use and investigate some common misunderstandings within the industry, with the aim of facilitating the appropriate selection and qualification of AMs. The key message we wish to emphasize is that in order to most effectively mitigate risk around cell therapy development and patient safety, users must work with their suppliers and regulators to qualify each AM to assess source, purity, identity, safety, and suitability in a given application. Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  1. Disaggregating census data for population mapping using random forests with remotely-sensed and ancillary data.

    Directory of Open Access Journals (Sweden)

    Forrest R Stevens

    Full Text Available High resolution, contemporary data on human population distributions are vital for measuring impacts of population growth, monitoring human-environment interactions and for planning and policy development. Many methods are used to disaggregate census data and predict population densities for finer scale, gridded population data sets. We present a new semi-automated dasymetric modeling approach that incorporates detailed census and ancillary data in a flexible, "Random Forest" estimation technique. We outline the combination of widely available, remotely-sensed and geospatial data that contribute to the modeled dasymetric weights and then use the Random Forest model to generate a gridded prediction of population density at ~100 m spatial resolution. This prediction layer is then used as the weighting surface to perform dasymetric redistribution of the census counts at a country level. As a case study we compare the new algorithm and its products for three countries (Vietnam, Cambodia, and Kenya with other common gridded population data production methodologies. We discuss the advantages of the new method and increases over the accuracy and flexibility of those previous approaches. Finally, we outline how this algorithm will be extended to provide freely-available gridded population data sets for Africa, Asia and Latin America.

  2. An Experimental Study of Energy Consumption in Buildings Providing Ancillary Services

    Energy Technology Data Exchange (ETDEWEB)

    Lin, Yashen [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Afshari, Sina [University of Michigan; Wolfe, John [University of Michigan; Nazir, Md Salman [University of Michigan; Hiskens, Ian A. [University of Michigan; Johnson, Jeremiah X. [University of Michigan; Mathieu, Johanna L. [University of Michigan; Barnes, Arthur K. [Los Alamos National Laboratory; Geller, Drew A. [Los Alamos National Laboratory; Backhaus, Scott N. [Los Alamos National Laboratory

    2017-10-03

    Heating, ventilation, and air conditioning (HVAC) systems in commercial buildings can provide ancillary services (AS) to the power grid, but by providing AS their energy consumption may increase. This inefficiency is evaluated using round-trip efficiency (RTE), which is defined as the ratio between the decrease and the increase in the HVAC system's energy consumption compared to the baseline consumption as a result of providing AS. This paper evaluates the RTE of a 30,000 m2 commercial building providing AS. We propose two methods to estimate the HVAC system's settling time after an AS event based on temperature and the air flow measurements from the building. Experimental data gathered over a 4-month period are used to calculate the RTE for AS signals of various waveforms, magnitudes, durations, and polarities. The results indicate that the settling time estimation algorithm based on the air flow measurements obtains more accurate results compared to the temperature-based algorithm. Further, we study the impact of the AS signal shape parameters on the RTE and discuss the practical implications of our findings.

  3. Thermal ice loads on dams and ancillary structures: A brief review

    International Nuclear Information System (INIS)

    Gerard, R.

    1989-01-01

    A major consideration in the design of low to medium head dams in cold regions is the thrust exerted by thermal expansion of a solid ice sheet. Such loads are also of concern in the design of gates, intakes and other ancillary structures. Such loads can be greater than 300-400 kilo Newtons per meter, and are of greatest concern when ice is unshielded by snow from temperature fluctuations. Details are presented of calculation of thermal ice loads, and field measurements of thermal ice forces. Past structural failures, field and laboratory investigations, and analyses, all confirm that thermal ice loads on wide structures such as dams, and isolated structures such as bridge piers and water intakes, can be much more significant than is suggested by the loads currently specified in various North American design guidelines for hydraulic structures. While some guidelines for thermal ice loads are excessively conservative, particularly for protected situations such as gates set between piers, in other more common situations they are dangerously low. Three useful approaches that would yield information for improving thermal ice load specification are: hindcast upper bounds on thermal ice loads by assessing the ice regime and load bearing capacity of existing structures; field measurement of thermal ice loads and stresses using modern instrumentation; and measurement and analysis of the formation and movement of lake and reservoir ice covers. 23 refs., 4 figs

  4. Analysis of the Effects of a Flexible Ramping Ancillary Service Product on Power System Operations: Preprint

    Energy Technology Data Exchange (ETDEWEB)

    Krad, Ibrahim; Ibanez, Eduardo; Ela, Erik

    2015-10-19

    The recent increased interest in utilizing variable generation (VG) resources such as wind and solar in power systems has motivated investigations into new operating procedures. Although these resources provide desirable value to a system (e.g., no fuel costs or emissions), interconnecting them provides unique challenges. Their variable, non-controllable nature in particular requires significant attention, because it directly results in increased power system variability and uncertainty. One way to handle this is via new operating reserve schemes. Operating reserves provide upward and downward generation and ramping capacity to counteract uncertainty and variability prior to their realization. For instance, uncertainty and variability in real-time dispatch can be accounted for in the hour-ahead unit commitment. New operating reserve methodologies that specifically account for the increased variability and uncertainty caused by VG are currently being investigated and developed by academia and industry. This paper examines one method inspired by the new operating reserve product being proposed by the California Independent System Operator. The method is based on examining the potential ramping requirements at any given time and enforcing those requirements via a reserve demand curve in the market-clearing optimization as an additional ancillary service product.

  5. Mid-term fire danger index based on satellite imagery and ancillary geographic data

    Science.gov (United States)

    Stefanidou, A.; Dragozi, E.; Tompoulidou, M.; Stepanidou, L.; Grigoriadis, D.; Katagis, T.; Stavrakoudis, D.; Gitas, I.

    2017-09-01

    Fire danger forecast constitutes one of the most important components of integrated fire management since it provides crucial information for efficient pre-fire planning, alertness and timely response to a possible fire event. The aim of this work is to develop an index that has the capability of predicting accurately fire danger on a mid-term basis. The methodology that is currently under development is based on an innovative approach that employs dry fuel spatial connectivity as well as biophysical and topological variables for the reliable prediction of fire danger. More specifically, the estimation of the dry fuel connectivity is based on a previously proposed automated procedure implemented in R software that uses Moderate Resolution Imaging Spectrometer (MODIS) time series data. Dry fuel connectivity estimates are then combined with other ancillary data such as fuel type and proximity to roads in order to result in the generation of the proposed mid-term fire danger index. The innovation of the proposed index—which will be evaluated by comparison to historical fire data—lies in the fact that its calculation is almost solely affected by the availability of satellite data. Finally, it should be noted that the index is developed within the framework of the National Observatory of Forest Fires (NOFFi) project.

  6. Impact of Production from Photovoltaic Power Plants on Increase of Ancillary Services in the Czech Republic

    Directory of Open Access Journals (Sweden)

    Martin Smocek

    2016-01-01

    Full Text Available Renewable energy resources represent a noticeable part of the overall energetic concept development. New integration of renewable energy resources into power grids has a significant impact on the reliability and quality of power supply. The major problem of the photovoltaic and wind power plants is their dependency on weather conditions, since it has a direct effect on their immediate output produced that shows stochastic behaviour. These stochastic outputs result in very adverse impacts on the power grid. Further development of these resources could lead to exceeding of the control and absorption abilities of the power grid. The power grid must be set in balance with respect to the production and consumption of electric power at any time. The operation of photovoltaic power plants impair keeping this balance. That has an adverse impact on the very operation and maintenance of network parameters within the extent required. This survey deals with analysis focused on operation of the photovoltaic power plants with respect to the increase of reserve power in ancillary services in the Czech Republic.

  7. Proposal for a change of contractor to operate the hostels, apartments and ancillary premises

    CERN Document Server

    2001-01-01

    The purpose of this document is to inform the Finance Committee of the final outcome of the negotiations with the firm to which the contract was adjudicated for the operation of the hostels, apartments and ancillary premises run by the CERN Housing Service and to request approval for the contract placed with the second lowest bidder. In September 2000, the Finance Committee agreed to the negotiation of a contract with VIENNA INTERNATIONAL HOTEL MANAGEMENT (AT) - SEREG (CH) (cf. CERN/FC/4311). For the reasons explained in this document, the Finance Committee is now invited to approve a contract with the consortium ISS MULTISERVICE (CH) - ISS GEBAEUDESERVICE (DE), the second lowest bidder, for an initial period of five years, from 1 January 2001, for a total amount of 7 206 000 Swiss francs, not subject to revision until 31 December 2001. The contract includes an option for two one-year extensions beyond the initial five-year period. The consortium has indicated the following distribution by country of the cont...

  8. Cost-Causation-Based Tariffs for Wind Ancillary Service Impacts: Preprint

    Energy Technology Data Exchange (ETDEWEB)

    Kirby, B.; Milligan, M.; Wan, Y.

    2006-06-01

    Conference paper discussing the integration cost of wind. Although specific tariffs for wind generation for ancillary services are uncommon, we anticipate that balancing authorities (control areas) and other entities will move toward such tariffs. Tariffs for regulation and imbalance services should be cost-based, recognize the relevant time scales that correspond with utility operational cycles, and properly allocate those costs to those entities that cause the balancing authority to incur the costs. In this paper, we present methods for separating wind's impact into regulation and load following (imbalance) time scales. We show that approximating these impacts with simpler methods can significantly distort cost causation and even cause confusion between the relevant time scales. We present results from NREL's wind data collection program to illustrate the dangers of linearly scaling wind resource data from small wind plants to approximate the wind resource data from large wind plants. Finally, we provide a framework for developing regulation and imbalance tariffs, we outline methods to begin examining contingency reserve requirements for wind plants, we provide guidance on the important characteristics to consider, and we provide hypothetical cases that the tariff can be tested against to determine whether the results are desired.

  9. Removal Action Workplan for the 105-DR and 105-F Building Interim Safe Storage Projects and Ancillary Buildings

    International Nuclear Information System (INIS)

    Rodovsky, T. J.

    1999-01-01

    This document is the removal action workplan (RAW) for the 105-DR and 105-F Reactor Buildings and ancillary facilities. These buildings and facilities are located in the 100-D/DR and 100-F Areas of the Hanford Site in Benton County, Washington, which is owned and operated by the U.S. Department of Energy (DOE). The 100 Areas (including 100-D/DR and 100-F Areas) of the Hanford Site were placed on the U.S. Environmental Protection Agency's (EPA) National Priorities List under the Comprehensive Environmental Response, Compensation,and Liability Act of 1980 (CERCLA). DOE has determined that hazardous substances in the 105-DR and 105-F Reactor Buildings and four ancillary facilities present a potential threat to human health or the environment DOE has also determined that a non-time critical removal action is warranted at these facilities

  10. International Society of Gynecological Pathologists (ISGyP) Endometrial Cancer Project: Guidelines From the Special Techniques and Ancillary Studies Group.

    Science.gov (United States)

    Cho, Kathleen R; Cooper, Kumarasen; Croce, Sabrina; Djordevic, Bojana; Herrington, Simon; Howitt, Brooke; Hui, Pei; Ip, Philip; Koebel, Martin; Lax, Sigurd; Quade, Bradley J; Shaw, Patricia; Vidal, August; Yemelyanova, Anna; Clarke, Blaise; Hedrick Ellenson, Lora; Longacre, Teri A; Shih, Ie-Ming; McCluggage, W Glenn; Malpica, Anais; Oliva, Esther; Parkash, Vinita; Matias-Guiu, Xavier

    2018-04-11

    The aim of this article is to propose guidelines and recommendations in problematic areas in pathologic reporting of endometrial carcinoma (EC) regarding special techniques and ancillary studies. An organizing committee designed a comprehensive survey with different questions related to pathologic features, diagnosis, and prognosis of EC that was sent to all members of the International Society of Gynecological Pathologists. The special techniques/ancillary studies group received 4 different questions to be addressed. Five members of the group reviewed the literature and came up with recommendations and an accompanying text which were discussed and agreed upon by all members of the group. Twelve different recommendations are made. They address the value of immunohistochemistry, ploidy, and molecular analysis for assessing prognosis in EC, the value of steroid hormone receptor analysis to predict response to hormone therapy, and parameters regarding applying immunohistochemistry and molecular tests for assessing mismatch deficiency in EC.

  11. Short-Term Output Variations in Wind Farms--Implications for Ancillary Services in the United States: Preprint

    Energy Technology Data Exchange (ETDEWEB)

    Cadogan, J. [U.S. Department of Energy (US); Milligan, M. [National Renewable Energy Laboratory (US); Wan, Y. [National Renewable Energy Laboratory (US); Kirby, B. [Oak Ridge National Laboratory (US)

    2001-09-21

    With the advent of competition in the electric power marketplace, this paper reviews changes that affect wind and other renewable energy technologies, and discusses the role of federal and state policies in the recent wind installations in the United States. In particular, it reviews the implications of ancillary service requirements on a wind farm and presents initial operating results of monitoring one Midwest wind farm. Under federal energy policy, each generator must purchase, or otherwise provide for, ancillary services, such as dispatch, regulation, operation reserve, voltage regulation, and scheduling required to move power to load. As a renewable technology that depends on the forces of nature, short-term output variations are inherently greater for a wind farm than for a gas-fired combined cycle or a supercritical coal-fired unit.

  12. INTRINSIC REGULATION OF HEMOGLOBIN EXPRESSION BY VARIABLE SUBUNIT INTERFACE STRENGTHS

    Science.gov (United States)

    Manning, James M.; Popowicz, Anthony M.; Padovan, Julio C.; Chait, Brian T.; Manning, Lois R.

    2012-01-01

    SUMMARY The expression of the six types of human hemoglobin subunits over time is currently considered to be regulated mainly by transcription factors that bind to upstream control regions of the gene (the “extrinsic” component of regulation). Here we describe how subunit pairing and further assembly to tetramers in the liganded state is influenced by the affinity of subunits for one another (the “intrinsic” component of regulation). The adult hemoglobin dimers have the strongest subunit interfaces and the embryonic hemoglobins are the weakest with fetal hemoglobins of intermediate strength, corresponding to the temporal order of their expression. These variable subunit binding strengths and the attenuating effects of acetylation contribute to the differences with which these hemoglobin types form functional O2-binding tetramers consistent with gene switching. PMID:22129306

  13. Engineering Task Plan for the Integrity Assessment Examination of Double Contained Receiver Tanks (DCRT) Catch Tanks and Ancillary facilities

    Energy Technology Data Exchange (ETDEWEB)

    BECKER, D.L.

    2000-05-23

    This Engineering Task Plan (ETP) presents the integrity assessment examination of three DCRTs, seven catch tanks, and two ancillary facilities located in the 200 East and West Areas of the Hanford Site. The integrity assessment examinations, as described in this ETP, will provide the necessary information to enable the independently qualified registered professional engineer (IQRPE) to assess the condition and integrity of these facilities. The plan is consistent with the Double-Shell Tank Waste Transfer Facilities Integrity Assessment Plan.

  14. Engineering Task Plan for the Integrity Assessment Examination of Double-Contained Receiver Tanks (DCRT), Catch Tanks and Ancillary facilities

    International Nuclear Information System (INIS)

    BECKER, D.L.

    2000-01-01

    This Engineering Task Plan (ETP) presents the integrity assessment examination of three DCRTs, seven catch tanks, and two ancillary facilities located in the 200 East and West Areas of the Hanford Site. The integrity assessment examinations, as described in this ETP, will provide the necessary information to enable the independently qualified registered professional engineer (IQRPE) to assess the condition and integrity of these facilities. The plan is consistent with the Double-Shell Tank Waste Transfer Facilities Integrity Assessment Plan

  15. Regulation of BK channels by auxiliary γ subunits

    Directory of Open Access Journals (Sweden)

    Jiyuan eZhang

    2014-10-01

    Full Text Available The large-conductance, calcium- and voltage-activated potassium (BK channel has the largest single-channel conductance among potassium channels and can be activated by both membrane depolarization and increases in intracellular calcium concentration. BK channels consist of pore-forming, voltage- and calcium-sensing α subunits, either alone or in association with regulatory subunits. BK channels are widely expressed in various tissues and cells including both excitable and non-excitable cells and display diverse biophysical and pharmacological characteristics. This diversity can be explained in part by posttranslational modifications and alternative splicing of the α subunit, which is encoded by a single gene, KCNMA1, as well as by tissue-specific β subunit modulation. Recently, a leucine-rich repeat-containing membrane protein, LRRC26, was found to interact with BK channels and cause an unprecedented large negative shift (~-140 mV in the voltage dependence of the BK channel activation. LRRC26 allows BK channels to open even at near-physiological calcium concentration and membrane voltage in non-excitable cells. Three LRRC26-related proteins, LRRC52, LRRC55, and LRRC38, were subsequently identified as BK channel modulators. These LRRC proteins are structurally and functionally distinct from the BK channel β subunits and were designated as γ subunits. The discovery of the γ subunits adds a new dimension to BK channel regulation and improves our understanding of the physiological functions of BK channels in various tissues and cell types. Unlike BK channel β subunits, which have been intensively investigated both mechanistically and physiologically, our understanding of the γ subunits is very limited at this stage. This article reviews the structure, modulatory mechanisms, physiological relevance, and potential therapeutic implications of γ subunits as they are currently understood.

  16. Map of risks for the implementation of radio-frequency identification: application of ancillaries in the University Hospital Jean Verdier.

    Science.gov (United States)

    Bertrand, E; Schlatter, J

    2010-01-01

    Ancillaries are surgical instruments, such as orthopedical instruments set for reconstruction of knee (a mounting arm...) used to implant or extract prosthesis. Their management involves the departments of sterilization and surgery as well as the suppliers. Such a long circuit exposes the instruments to potential risk hazards like a lack of traceability as the suspicion of Creutzfeldt-Jakob. In order to reduce the risk of errors we will propose the implementation of radio-frequency identification (RFID) to trace the ancillaries during each step of the supply chain. The objective of our study is to analyze and to map the risks associated with RFID implementation. A preliminary analysis of risks (APR) is conducted to map out the hazards for the implementation of RFID. The APR identifies 162 scenarios with a maximum risk connected to environment and technology. To reduce the risks identified, 22 courses of action are proposed, such as audits, training, and internal controls. For each action, a procedure has been designed and evaluated. This preliminary analysis of risks allows targeting the potential dangers for the RFID implementation applied to ancillaries and reduces them significantly.

  17. Breaking through the hydrogen cost barrier by using electrolysis loads to access ancillary services and demand response programs

    International Nuclear Information System (INIS)

    Wilson, D.; McGillivray, R.

    2009-01-01

    This presentation described the use of hydrogen electrolysis as a load resource for handling grid instability resulting from the increased penetration of intermittent renewable power. In particular, it focused on Hydrogenics, the leading global supplier of industrial scale electrolysis equipment and fuel cells. The presentation included an overview of the current incentive and market value of ancillary services provided by the company and demand responses in a number of grids around the world. There is a link between the amount of ancillary services required by the grid and the penetration level of renewable energy power such as wind and solar. The ability of hydrogen generation from electrolysis to satisfy all the requirements of ancillary services markets was also demonstrated. The economic analysis of hydrogen generation was discussed with particular reference to the cost of hydrogen fully loading all capital, energy and operating costs. The resulting reduction in the cost of hydrogen was compared to the existing markets for hydrogen, including use of hydrogen as a fuel for municipal bus fleets relative to the existing cost of fossil fuel fleets. Current industrial hydrogen merchant and bulk market prices were also compared

  18. New alternatives for reference evapotranspiration estimation in West Africa using limited weather data and ancillary data supply strategies.

    Science.gov (United States)

    Landeras, Gorka; Bekoe, Emmanuel; Ampofo, Joseph; Logah, Frederick; Diop, Mbaye; Cisse, Madiama; Shiri, Jalal

    2018-05-01

    Accurate estimation of reference evapotranspiration ( ET 0 ) is essential for the computation of crop water requirements, irrigation scheduling, and water resources management. In this context, having a battery of alternative local calibrated ET 0 estimation methods is of great interest for any irrigation advisory service. The development of irrigation advisory services will be a major breakthrough for West African agriculture. In the case of many West African countries, the high number of meteorological inputs required by the Penman-Monteith equation has been indicated as constraining. The present paper investigates for the first time in Ghana, the estimation ability of artificial intelligence-based models (Artificial Neural Networks (ANNs) and Gene Expression Programing (GEPs)), and ancillary/external approaches for modeling reference evapotranspiration ( ET 0 ) using limited weather data. According to the results of this study, GEPs have emerged as a very interesting alternative for ET 0 estimation at all the locations of Ghana which have been evaluated in this study under different scenarios of meteorological data availability. The adoption of ancillary/external approaches has been also successful, moreover in the southern locations. The interesting results obtained in this study using GEPs and some ancillary approaches could be a reference for future studies about ET 0 estimation in West Africa.

  19. MODIS/Aqua 0.5 x 0.5 degree Aeronet cutouts of MODIS L1B Radiances, Geolocation, Cloud Mask, and ancillary data

    Data.gov (United States)

    National Aeronautics and Space Administration — The MYDARNSS data set consists of MODIS Atmosphere and Ancillary Products subsets that are generated over a number of Aerosol Robotic Network (AERONET) sites. These...

  20. MODIS/Terra 0.5 x 0.5 degree FluxNet cutouts of MODIS L1B Radiances, Geolocation, Cloud Mask, and ancillary data

    Data.gov (United States)

    National Aeronautics and Space Administration — The MODFNSS data set consists of MODIS Atmosphere and Ancillary Products subsets that are generated over a number of FLUXNET sites. FLUXNET is a network of...

  1. MODIS/Aqua 0.5 x 0.5 degree FluxNet cutouts of MODIS L1B Radiances, Geolocation, Cloud Mask, and ancillary data

    Data.gov (United States)

    National Aeronautics and Space Administration — The MYDFNSS data set consists of MODIS Atmosphere and Ancillary Products subsets that are generated over a number of FLUXNET sites. FLUXNET is a network of...

  2. LANDMET Ancillary Monthly Mean Thermal Effective Infrared and Microwave Emissivity Data L3 V1 (LANDMET_ANC_TEIME) at GES DISC

    Data.gov (United States)

    National Aeronautics and Space Administration — This is an ancillary product, climatology monthly mean and standard deviation, containing a number of emissivity data. They are broadband thermal IR emissivity from...

  3. Regional land cover characterization using Landsat thematic mapper data and ancillary data sources

    Science.gov (United States)

    Vogelmann, James E.; Sohl, Terry L.; Campbell, P.V.; Shaw, D.M.; ,

    1998-01-01

    As part of the activities of the Multi-Resolution Land Characteristics (MRLC) Interagency Consortium, an intermediate-scale land cover data set is being generated for the conterminous United States. This effort is being conducted on a region-by-region basis using U.S. Standard Federal Regions. To date, land cover data sets have been generated for Federal Regions 3 (Pennsylvania, West Virginia, Virginia, Maryland, and Delaware) and 2 (New York and New Jersey). Classification work is currently under way in Federal Region 4 (the southeastern United States), and land cover mapping activities have been started in Federal Regions 5 (the Great Lakes region) and 1 (New England). It is anticipated that a land cover data set for the conterminous United States will be completed by the end of 1999. A standard land cover classification legend is used, which is analogous to and compatible with other classification schemes. The primary MRLC regional classification scheme contains 23 land cover classes.The primary source of data for the project is the Landsat thematic mapper (TM) sensor. For each region, TM scenes representing both leaf-on and leaf-off conditions are acquired, preprocessed, and georeferenced to MRLC specifications. Mosaicked data are clustered using unsupervised classification, and individual clusters are labeled using aerial photographs. Individual clusters that represent more than one land cover unit are split using spatial modeling with multiple ancillary spatial data layers (most notably, digital elevation model, population, land use and land cover, and wetlands information). This approach yields regional land cover information suitable for a wide array of applications, including landscape metric analyses, land management, land cover change studies, and nutrient and pesticide runoff modeling.

  4. The validation of corrosion inhibitors for ancillary cooling systems at Sizewell B PWR

    International Nuclear Information System (INIS)

    Allan, S.J.; Garnsey, R.; Lawson, W.F.

    1994-01-01

    Sizewell B has several low temperature ancillary cooling circuits for removing low grade heat. One of the principal circuits is the Component Cooling Water System (CCWS) primarily constructed from carbon steel pipework but also containing stainless steel, titanium, copper and cupronickel components. Linked to the CCWS is another carbon steel system namely the Reserve Ultimate Heat Sink (RUHS) a system unique to Sizewell B providing an independent heat removal capability for the CCWS as a safety back-up. Both systems utilize demineralized water with the addition of an inhibitor to prevent corrosion. The vendor recommended inhibitor namely potassium chromate is highly toxic to marine life and initial discussions with the regulatory authorities indicated that it would be extremely difficult to obtain a discharge consent to cover leakage or potential system purges and dumps associated with maintenance operations. Thus an alternative was sought. Sodium nitrite (anhydrous NaN0 2 )/borax (Na 2 B 4 0 7 1OH 2 0) inhibitor solutions have been used for many years in the power industry for preservation of carbon steel plant and several proprietary formulations are based on this mixture. There has been extensive test work in the UK on this inhibitor for secondary side boiler storage on gas reactors and therefore it was considered a prime candidate for use at Sizewell B. Starting in 1985 a programme of work sponsored by Nuclear Electric plc has been completed by NNC Ltd to compare nitrite/borax with other suitable inhibitors and validate its use for Sizewell B. (authors). 5 figs., 1 tab

  5. Investigating the Impact of Climate Change on Hydroelectric Generation and Ancillary Services in California

    Science.gov (United States)

    Forrest, K.; Tarroja, B.; AghaKouchak, A.; Chiang, F.; Samuelsen, S.

    2017-12-01

    Spatial and temporal shifts in hydrological regimes predicted under climate change conditions have implications for the management of reservoirs and hydropower contributions to generation and ancillary services. California relies on large hydropower plants to provide flexible electricity generation, which will be increasingly important for supporting renewable resources. This study examines the impact of climate change on large hydropower generation in California. Four climate models for Representative Concentration Pathways (RCP) 4.5 and RCP 8.5 are utilized to evaluate the impact of the climate change conditions on (1) the magnitude and profile of hydropower generation and (2) the ability of hydropower to provide spinning reserve. Under both RCP scenarios, impacts are regionally dependent, with precipitation projected to increase in northern California and decrease in southern California for the ten-year period investigated (2046-2055). The overall result is a net increase in inflow into large hydropower units as a majority of the hydropower plants studied are located in the northern part of the state. Increased inflow is primarily driven by increased runoff during the winter and does not necessarily result in increased generation, as extreme events yield greater overall spillage, up to 45% of total inflow. Increased winter hydropower generation paired with increased reservoir constraints in summer result in an 11 to 18% decrease in spinning reserve potential across the year. Under high inflow conditions there is a decreased flexibility for choosing generation versus spinning reserve as water needs to be released, regardless. During summer, hydropower units providing spinning reserve experienced decreased inflow and lower reservoir levels compared to the historical baseline, resulting in decreased spinning reserve bidding potential. Decreased bidding, especially during summer periods at peak electricity demand, can result in greater demand for other dispatchable

  6. Camera traps and mark-resight models: The value of ancillary data for evaluating assumptions

    Science.gov (United States)

    Parsons, Arielle W.; Simons, Theodore R.; Pollock, Kenneth H.; Stoskopf, Michael K.; Stocking, Jessica J.; O'Connell, Allan F.

    2015-01-01

    Unbiased estimators of abundance and density are fundamental to the study of animal ecology and critical for making sound management decisions. Capture–recapture models are generally considered the most robust approach for estimating these parameters but rely on a number of assumptions that are often violated but rarely validated. Mark-resight models, a form of capture–recapture, are well suited for use with noninvasive sampling methods and allow for a number of assumptions to be relaxed. We used ancillary data from continuous video and radio telemetry to evaluate the assumptions of mark-resight models for abundance estimation on a barrier island raccoon (Procyon lotor) population using camera traps. Our island study site was geographically closed, allowing us to estimate real survival and in situ recruitment in addition to population size. We found several sources of bias due to heterogeneity of capture probabilities in our study, including camera placement, animal movement, island physiography, and animal behavior. Almost all sources of heterogeneity could be accounted for using the sophisticated mark-resight models developed by McClintock et al. (2009b) and this model generated estimates similar to a spatially explicit mark-resight model previously developed for this population during our study. Spatially explicit capture–recapture models have become an important tool in ecology and confer a number of advantages; however, non-spatial models that account for inherent individual heterogeneity may perform nearly as well, especially where immigration and emigration are limited. Non-spatial models are computationally less demanding, do not make implicit assumptions related to the isotropy of home ranges, and can provide insights with respect to the biological traits of the local population.

  7. Echinococcus granulosus Antigen B Structure: Subunit Composition and Oligomeric States

    Science.gov (United States)

    Monteiro, Karina M.; Cardoso, Mateus B.; Follmer, Cristian; da Silveira, Nádya P.; Vargas, Daiani M.; Kitajima, Elliot W.; Zaha, Arnaldo; Ferreira, Henrique B.

    2012-01-01

    Background Antigen B (AgB) is the major protein secreted by the Echinococcus granulosus metacestode and is involved in key host-parasite interactions during infection. The full comprehension of AgB functions depends on the elucidation of several structural aspects that remain unknown, such as its subunit composition and oligomeric states. Methodology/Principal Findings The subunit composition of E. granulosus AgB oligomers from individual bovine and human cysts was assessed by mass spectrometry associated with electrophoretic analysis. AgB8/1, AgB8/2, AgB8/3 and AgB8/4 subunits were identified in all samples analyzed, and an AgB8/2 variant (AgB8/2v8) was found in one bovine sample. The exponentially modified protein abundance index (emPAI) was used to estimate the relative abundance of the AgB subunits, revealing that AgB8/1 subunit was relatively overrepresented in all samples. The abundance of AgB8/3 subunit varied between bovine and human cysts. The oligomeric states formed by E. granulosus AgB and recombinant subunits available, rAgB8/1, rAgB8/2 and rAgB8/3, were characterized by native PAGE, light scattering and microscopy. Recombinant subunits showed markedly distinct oligomerization behaviors, forming oligomers with a maximum size relation of rAgB8/3>rAgB8/2>rAgB8/1. Moreover, the oligomeric states formed by rAgB8/3 subunit were more similar to those observed for AgB purified from hydatid fluid. Pressure-induced dissociation experiments demonstrated that the molecular assemblies formed by the more aggregative subunits, rAgB8/2 and rAgB8/3, also display higher structural stability. Conclusions/Significance For the first time, AgB subunit composition was analyzed in samples from single hydatid cysts, revealing qualitative and quantitative differences between samples. We showed that AgB oligomers are formed by different subunits, which have distinct abundances and oligomerization properties. Overall, our findings have significantly contributed to increase the

  8. Transcriptional regulators of Na, K-ATPase subunits

    Directory of Open Access Journals (Sweden)

    Zhiqin eLi

    2015-10-01

    Full Text Available The Na,K-ATPase classically serves as an ion pump creating an electrochemical gradient across the plasma membrane that is essential for transepithelial transport, nutrient uptake and membrane potential. In addition, Na,K-ATPase also functions as a receptor, a signal transducer and a cell adhesion molecule. With such diverse roles, it is understandable that the Na,K-ATPase subunits, the catalytic alpha-subunit, the beta-subunit and the FXYD proteins, are controlled extensively during development and to accommodate physiological needs. The spatial and temporal expression of Na,K-ATPase is partially regulated at the transcriptional level. Numerous transcription factors, hormones, growth factors, lipids and extracellular stimuli modulate the transcription of the Na,K-ATPase subunits. Moreover, epigenetic mechanisms also contribute to the regulation of Na,K-ATPase expression. With the ever growing knowledge about diseases associated with the malfunction of Na,K-ATPase, this review aims at summarizing the best-characterized transcription regulators that modulate Na,K-ATPase subunit levels. As abnormal expression of Na,K-ATPase subunits have been observed in many carcinoma, we will also discuss transcription factors that are associated with epithelial-to-mesenchymal transition, a crucial step in the progression of many tumors to malignant disease.

  9. Impact of Market Behavior, Fleet Composition, and Ancillary Services on Revenue Sufficiency

    Energy Technology Data Exchange (ETDEWEB)

    Frew, Bethany [National Renewable Energy Lab. (NREL), Golden, CO (United States); Gallo, Giulia [National Renewable Energy Lab. (NREL), Golden, CO (United States); Brinkman, Gregory [National Renewable Energy Lab. (NREL), Golden, CO (United States); Milligan, Michael [National Renewable Energy Lab. (NREL), Golden, CO (United States); Clark, Kara [National Renewable Energy Lab. (NREL), Golden, CO (United States); Bloom, Aaron [National Renewable Energy Lab. (NREL), Golden, CO (United States)

    2016-06-01

    Revenue insufficiency, or the missing money problem, occurs when the revenues that generators earn from the market are not sufficient to cover both fixed and variable costs to remain in the market and/or justify investments in new capacity, which may be needed for reliability. The near-zero marginal cost of variable renewable generators further exacerbates these revenue challenges. Estimating the extent of the missing money problem in current electricity markets is an important, nontrivial task that requires representing both how the power system operates and how market participants behave. This paper explores the missing money problem using a production cost model that represented a simplified version of the Electric Reliability Council of Texas (ERCOT) energy-only market for the years 2012-2014. We evaluate how various market structures -- including market behavior, ancillary services, and changing fleet compositions -- affect net revenues in this ERCOT-like system. In most production cost modeling exercises, resources are assumed to offer their marginal capabilities at marginal costs. Although this assumption is reasonable for feasibility studies and long-term planning, it does not adequately consider the market behaviors that impact revenue sufficiency. In this work, we simulate a limited set of market participant strategic bidding behaviors by means of different sets of markups; these markups are applied to the true production costs of all gas generators, which are the most prominent generators in ERCOT. Results show that markups can help generators increase their net revenues overall, although net revenues may increase or decrease depending on the technology and the year under study. Results also confirm that conventional, variable-cost-based production cost simulations do not capture prices accurately, and this particular feature calls for proxies for strategic behaviors (e.g., markups) and more accurate representations of how electricity markets work. The

  10. From a bundled energy-capacity pricing model to an energy-capacity-ancillary services pricing model

    International Nuclear Information System (INIS)

    Raineri, Ricardo; Arce, Raul; Rios, Sebastian; Salamanca, Carlos

    2008-01-01

    In this paper, we extend the Chilean power generation pricing mechanism, with capacity and energy payments, to one where ancillary services (AS), as frequency regulation and voltage control, are explicitly recognized. Adequacy and security attributes of the electric system and the public good characteristics of AS are set within the payment structure to distribute the financing of AS among those who benefit from their provision. The contribution to finance the provision of AS is determined assessing the value assigned to the system security by each agent, following what's an efficient pricing mechanism in the presence of public goods

  11. Strategies for carbon dioxide emissions reductions: Residential natural gas efficiency, economic, and ancillary health impacts in Maryland

    International Nuclear Information System (INIS)

    Ruth, Matthias; Blohm, Andrew; Mauer, Joanna; Gabriel, Steven A.; Kesana, Vijay G.; Chen Yihsu; Hobbs, Benjamin F.; Irani, Daraius

    2010-01-01

    As part of its commitments to the Regional Greenhouse Gas Initiative (RGGI), the State of Maryland, USA, auctions emission permits to electric utilities, creating revenue that can be used to benefit consumers and the environment. This paper explores the CO 2 emissions reductions that may be possible by allocating some of that revenue to foster efficiency improvements in the residential sector's use of natural gas. Since these improvements will require changes to the capital stock of houses and end use equipment, efficiency improvements may be accompanied by economic and ancillary health impacts, both of which are quantified in this paper.

  12. Phosphorescent Pt(ii) complexes bearing a monoanionic C^N^N luminophore and tunable ancillary ligands.

    Science.gov (United States)

    Hebenbrock, Marian; Stegemann, Linda; Kösters, Jutta; Doltsinis, Nikos L; Müller, Jens; Strassert, Cristian A

    2017-03-07

    A versatile design strategy is presented towards new monoanionic pincer luminophores, showing that cyclometallating C^N^N ligands can yield phosphorescent Pt(ii) complexes even if a neutral 1,2,3-triazole ring is inserted by click chemistry. The overall charge, intermolecular interactions and excited state properties can be manipulated and controlled by varying the nature of the ancillary ligand, and its effect on the structural and the triplet state characteristics can be thoroughly investigated and correlated by means of theory and spectroscopy.

  13. Pituitary glycoprotein hormone a-subunit secretion by cirrhotic patients

    Directory of Open Access Journals (Sweden)

    Oliveira M.C.

    1999-01-01

    Full Text Available Secretion of the a-subunit of pituitary glycoprotein hormones usually follows the secretion of intact gonadotropins and is increased in gonadal failure and decreased in isolated gonadotropin deficiency. The aim of the present study was to determine the levels of the a-subunit in the serum of patients with cirrhosis of the liver and to compare the results obtained for eugonadal cirrhotic patients with those obtained for cirrhotic patients with hypogonadotropic hypogonadism. Forty-seven of 63 patients with cirrhosis (74.6% presented hypogonadism (which was central in 45 cases and primary in 2, 7 were eugonadal, and 9 women were in normal menopause. The serum a-subunit was measured by the fluorimetric method using monoclonal antibodies. Cross-reactivity with LH, TSH, FSH and hCG was 6.5, 1.2, 4.3 and 1.1%, respectively, with an intra-assay coefficient of variation (CV of less than 5% and an interassay CV of 5%, and sensitivity limit of 4 ng/l. The serum a-subunit concentration ranged from 36 to 6253 ng/l, with a median of 273 ng/l. The median was 251 ng/l for patients with central hypogonadism and 198 ng/l for eugonadal patients. The correlation between the a-subunit and basal LH levels was significant both in the total sample (r = 0.48, P<0.01 and in the cirrhotic patients with central hypogonadism (r = 0.33, P = 0.02. Among men with central hypogonadism there was a negative correlation between a-subunit levels and total testosterone levels (r = 0.54, P<0.01 as well as free testosterone levels (r = -0.53, P<0.01. In conclusion, although the a-subunit levels are correlated with LH levels, at present they cannot be used as markers for hypogonadism in patients with cirrhosis of the liver.

  14. Modes of Operation of the BKCa Channel β2 Subunit

    Science.gov (United States)

    Savalli, Nicoletta; Kondratiev, Andrei; de Quintana, Sarah Buxton; Toro, Ligia; Olcese, Riccardo

    2007-01-01

    The β2 subunit of the large conductance Ca2+- and voltage-activated K+ channel (BKCa) modulates a number of channel functions, such as the apparent Ca2+/voltage sensitivity, pharmacological and kinetic properties of the channel. In addition, the N terminus of the β2 subunit acts as an inactivating particle that produces a relatively fast inactivation of the ionic conductance. Applying voltage clamp fluorometry to fluorescently labeled human BKCa channels (hSlo), we have investigated the mechanisms of operation of the β2 subunit. We found that the leftward shift on the voltage axis of channel activation curves (G(V)) produced by coexpression with β2 subunits is associated with a shift in the same direction of the fluorescence vs. voltage curves (F(V)), which are reporting the voltage dependence of the main voltage-sensing region of hSlo (S4-transmembrane domain). In addition, we investigated the inactivating mechanism of the β2 subunits by comparing its properties with the ones of the typical N-type inactivation process of Shaker channel. While fluorescence recordings from the inactivated Shaker channels revealed the immobilization of the S4 segments in the active conformation, we did not observe a similar feature in BKCa channels coexpressed with the β2 subunit. The experimental observations are consistent with the view that the β2 subunit of BKCa channels facilitates channel activation by changing the voltage sensor equilibrium and that the β2-induced inactivation process does not follow a typical N-type mechanism. PMID:17591990

  15. Modes of operation of the BKCa channel beta2 subunit.

    Science.gov (United States)

    Savalli, Nicoletta; Kondratiev, Andrei; de Quintana, Sarah Buxton; Toro, Ligia; Olcese, Riccardo

    2007-07-01

    The beta(2) subunit of the large conductance Ca(2+)- and voltage-activated K(+) channel (BK(Ca)) modulates a number of channel functions, such as the apparent Ca(2+)/voltage sensitivity, pharmacological and kinetic properties of the channel. In addition, the N terminus of the beta(2) subunit acts as an inactivating particle that produces a relatively fast inactivation of the ionic conductance. Applying voltage clamp fluorometry to fluorescently labeled human BK(Ca) channels (hSlo), we have investigated the mechanisms of operation of the beta(2) subunit. We found that the leftward shift on the voltage axis of channel activation curves (G(V)) produced by coexpression with beta(2) subunits is associated with a shift in the same direction of the fluorescence vs. voltage curves (F(V)), which are reporting the voltage dependence of the main voltage-sensing region of hSlo (S4-transmembrane domain). In addition, we investigated the inactivating mechanism of the beta(2) subunits by comparing its properties with the ones of the typical N-type inactivation process of Shaker channel. While fluorescence recordings from the inactivated Shaker channels revealed the immobilization of the S4 segments in the active conformation, we did not observe a similar feature in BK(Ca) channels coexpressed with the beta(2) subunit. The experimental observations are consistent with the view that the beta(2) subunit of BK(Ca) channels facilitates channel activation by changing the voltage sensor equilibrium and that the beta(2)-induced inactivation process does not follow a typical N-type mechanism.

  16. NOX Activation by Subunit Interaction and Underlying Mechanisms in Disease.

    Science.gov (United States)

    Rastogi, Radhika; Geng, Xiaokun; Li, Fengwu; Ding, Yuchuan

    2016-01-01

    Nicotinamide adenine dinucleotide phosphate (NAPDH) oxidase (NOX) is an enzyme complex with the sole function of producing superoxide anion and reactive oxygen species (ROS) at the expense of NADPH. Vital to the immune system as well as cellular signaling, NOX is also involved in the pathologies of a wide variety of disease states. Particularly, it is an integral player in many neurological diseases, including stroke, TBI, and neurodegenerative diseases. Pathologically, NOX produces an excessive amount of ROS that exceed the body's antioxidant ability to neutralize them, leading to oxidative stress and aberrant signaling. This prevalence makes it an attractive therapeutic target and as such, NOX inhibitors have been studied and developed to counter NOX's deleterious effects. However, recent studies of NOX have created a better understanding of the NOX complex. Comprised of independent cytosolic subunits, p47- phox , p67- phox , p40- phox and Rac , and membrane subunits, gp91- phox and p22- phox , the NOX complex requires a unique activation process through subunit interaction. Of these subunits, p47- phox plays the most important role in activation, binding and translocating the cytosolic subunits to the membrane and anchoring to p22- phox to organize the complex for NOX activation and function. Moreover, these interactions, particularly that between p47- phox and p22- phox , are dependent on phosphorylation initiated by upstream processes involving protein kinase C (PKC). This review will look at these interactions between subunits and with PKC. It will focus on the interaction involving p47- phox with p22- phox , key in bringing the cytosolic subunits to the membrane. Furthermore, the implication of these interactions as a target for NOX inhibitors such as apocynin will be discussed as a potential avenue for further investigation, in order to develop more specific NOX inhibitors based on the inhibition of NOX assembly and activation.

  17. Nicotinic acetylcholine receptor subunits in rhesus monkey retina.

    Science.gov (United States)

    Liu, Ji; McGlinn, Alice M; Fernandes, Alcides; Milam, Ann H; Strang, Christianne E; Andison, Margot E; Lindstrom, Jon M; Keyser, Kent T; Stone, Richard A

    2009-03-01

    The purpose of this study was to detect and establish the cellular localizations of nicotinic acetylcholine receptor (nAChR) subunits in Rhesus monkey retina. Retinas were dissected from the eyes of monkeys killed after unrelated experiments. RNA was extracted and analyzed by RT-PCR, using primers designed against human sequences of alpha3-alpha7, alpha9, and beta2-beta4 nAChR subunits. The RT-PCR products were separated by gel electrophoresis and sequenced. Frozen sections of postmortem fixed monkey eyes were immunolabeled with well-characterized and specific monoclonal antibodies against the alpha3, alpha4, alpha6, alpha7, beta2, or beta4 nAChR subunits and visualized with fluorescence labeling. Products of the predicted size for the alpha3-alpha7, alpha9, and beta2-beta4 nAChR subunits were detected by RT-PCR in Rhesus monkey retina. Homology between transcripts from monkey retina and human nucleotide sequences ranged from 93 to 99%. Immunohistochemical studies demonstrated that neurons in various cell layers of monkey retina expressed alpha3, alpha4, alpha7, or beta2 nAChR subunits and cells with the morphology of microglia were immunoreactive for the alpha6 or beta4 nAChR subunits. nAChR subunits are expressed in the monkey retina and localize to diverse retinal neurons as well as putative microglia. Besides mediating visual processing, retinal nAChRs may influence refractive development and ocular pathologies such as neovascularization.

  18. A bioinformatic and computational study of myosin phosphatase subunit diversity.

    Science.gov (United States)

    Dippold, Rachael P; Fisher, Steven A

    2014-08-01

    Variability in myosin phosphatase (MP) subunits may provide specificity in signaling pathways that regulate muscle tone. We utilized public databases and computational algorithms to investigate the phylogenetic diversity of MP regulatory (PPP1R12A-C) and inhibitory (PPP1R14A-D) subunits. The comparison of exonic coding sequences and expression data confirmed or refuted the existence of isoforms and their tissue-specific expression in different model organisms. The comparison of intronic and exonic sequences identified potential expressional regulatory elements. As examples, smooth muscle MP regulatory subunit (PPP1R12A) is highly conserved through evolution. Its alternative exon E24 is present in fish through mammals with two invariant features: 1) a reading frame shift generating a premature termination codon and 2) a hexanucleotide sequence adjacent to the 3' splice site hypothesized to be a novel suppressor of exon splicing. A characteristic of the striated muscle MP regulatory subunit (PPP1R12B) locus is numerous and phylogenetically variable transcriptional start sites. In fish this locus only codes for the small (M21) subunit, suggesting the primordial function of this gene. Inhibitory subunits show little intragenic variability; their diversity is thought to have arisen by expansion and tissue-specific expression of different gene family members. We demonstrate differences in the regulatory landscape between smooth muscle enriched (PPP1R14A) and more ubiquitously expressed (PPP1R14B) family members and identify deeply conserved intronic sequence and predicted transcriptional cis-regulatory elements. This bioinformatic and computational study has uncovered a number of attributes of MP subunits that supports selection of ideal model organisms and testing of hypotheses regarding their physiological significance and regulated expression. Copyright © 2014 the American Physiological Society.

  19. Photocatalytic Conversion of CO2 to CO using Rhenium Bipyridine Platforms Containing Ancillary Phenyl or BODIPY Moieties

    Science.gov (United States)

    Andrade, Gabriel A.; Pistner, Allen J.; Yap, Glenn P.A.; Lutterman, Daniel A.; Rosenthal, Joel

    2013-01-01

    Harnessing of solar energy to drive the reduction of carbon dioxide to fuels requires the development of efficient catalysts that absorb sunlight. In this work, we detail the synthesis, electrochemistry and photophysical properties of a set of homologous fac-ReI(CO)3 complexes containing either an ancillary phenyl (8) or BODIPY (12) substituent. These studies demonstrate that both the electronic properties of the rhenium center and BODIPY chromophore are maintained for these complexes. Photolysis studies demonstrate that both assemblies 8 and 12 are competent catalysts for the photochemical reduction of CO2 to CO in DMF using triethanolamine (TEOA) as a sacrificial reductant. Both compounds 8 and 12 display TOFs for photocatalytic CO production upon irradiation with light (λex ≥ 400 nm) of ~5 hr−1 with TON values of approximately 20. Although structural and photophysical measurements demonstrate that electronic coupling between the BODIPY and fac-ReI(CO)3 units is limited for complex 12, this work clearly shows that the photoactive BODIPY moiety is tolerated during catalysis and does not interfere with the observed photochemistry. When taken together, these results provide a clear roadmap for the development of advanced rhenium bipyridine complexes bearing ancillary BODIPY groups for the efficient photocatalytic reduction of CO2 using visible light. PMID:24015374

  20. Effects of beta-adrenergic blockers with different ancillary properties on lipid peroxidation in hyperthyroid rat cardiac muscle.

    Science.gov (United States)

    Asayama, K; Dobashi, K; Hayashibe, H; Kato, K

    1989-10-01

    To determine whether beta-blockade protects rat heart against thyroxine (T4)-induced accelelation of lipid peroxidation, in vivo effects of 3 beta-blockers with different ancillary properties on the mitochondrial oxidative enzyme, antioxidant enzymes and lipid peroxide were investigated. The rats were rendered hyperthyroid by adding T4 to their drinking water for 3 weeks and were treated simultaneously with either carteolol (a blocker with partial agonist activity; 30 mg/kg/day), atenolol (50 mg/kg/day) or arotinolol (a blocker with weak alpha-blocking action; 50 mg/kg/day). The T4-induced tachycardia was alleviated completely by either atenolol or arotinolol, but only partially by carteolol. Cytochrome c oxidase activity in the heart muscle was increased by T4 with a parallel increase in manganese (mitochondrial) superoxide dismutase. Atenolol, but neither carteolol nor arotinolol, suppressed this increase. Similarly, the T4-induced acceleration of lipid peroxidation was suppressed by atenolol alone. Glutathione peroxidase was markedly decreased, and both copper zinc (cytosolic) superoxide dismutase and catalase were also decreased or tended to be decreased by T4. The levels of these 3 enzymes were only minimally affected by the beta-blocker treatments. These results suggest that beta-blockade suppresses mitochondrial hypermetabolism and protects heart muscle against oxidative stress in hyperthyroidism, and that the ancillary properties of beta-blockers such as partial agonist activity and alpha-blocking action negate the protection.

  1. Fine needle biopsy with cytology in paediatrics: the importance of a multidisciplinary approach and the role of ancillary techniques.

    Science.gov (United States)

    Barroca, H; Bom-Sucesso, M

    2014-02-01

    Fine needle biopsy (FNB) with cytology has long been regarded as an excellent technique as the first choice for diagnosing adult tumours. Being an inexpensive minimally invasive technique with high accuracy and diagnostic immediacy through rapid on-site evaluation, it is also ideal for implementation in the paediatric setting, particularly in developing countries. Furthermore, it allows complementary and advanced procedures such as flow cytometry, polymerase chain reaction (PCR) or fluorescence in situ hybridization (FISH), among others, which enhances the diagnostic capacity of this technique and gives it a key role in risk stratification and therapeutic decision-making for several tumours. The advantages of FNB are optimized in the setting of a multidisciplinary team where cytologist, clinician and radiologist play leading roles. Paediatric tumours are rare and most ancillary techniques are cost-effective but complex to be implemented in small centres with limited experience in paediatric pathology. Therefore reference centres are essential, in order to establish teams with extensive experience and expertise. Hence, any child with a suspected malignancy should be directly referred to a paediatric oncology unit. Focusing on a practical approach to the assessment of paediatric lymphadenopathies and non-central nervous system solid tumours we review the effectiveness of FNB as applied concurrently with ancillary techniques in a multidisciplinary approach to the diagnosis, prognosis and therapeutic decisions of paediatric tumours and tumour-like lesions. © 2013 John Wiley & Sons Ltd.

  2. On combining multi-normalization and ancillary measures for the optimal score level fusion of fingerprint and voice biometrics

    Science.gov (United States)

    Mohammed Anzar, Sharafudeen Thaha; Sathidevi, Puthumangalathu Savithri

    2014-12-01

    In this paper, we have considered the utility of multi-normalization and ancillary measures, for the optimal score level fusion of fingerprint and voice biometrics. An efficient matching score preprocessing technique based on multi-normalization is employed for improving the performance of the multimodal system, under various noise conditions. Ancillary measures derived from the feature space and the score space are used in addition to the matching score vectors, for weighing the modalities, based on their relative degradation. Reliability (dispersion) and the separability (inter-/intra-class distance and d-prime statistics) measures under various noise conditions are estimated from the individual modalities, during the training/validation stage. The `best integration weights' are then computed by algebraically combining these measures using the weighted sum rule. The computed integration weights are then optimized against the recognition accuracy using techniques such as grid search, genetic algorithm and particle swarm optimization. The experimental results show that, the proposed biometric solution leads to considerable improvement in the recognition performance even under low signal-to-noise ratio (SNR) conditions and reduces the false acceptance rate (FAR) and false rejection rate (FRR), making the system useful for security as well as forensic applications.

  3. Secure provision of reactive power ancillary services in competitive electricity markets

    Science.gov (United States)

    El-Samahy, Ismael

    decoupled and assumed to be fixed during the reactive power dispatch procedures; however, the effect of reactive power on real power is considered in the model by calculating the required reduction in real power output of a generator due to an increase in its reactive power supply. In this case, real power generation is allowed to be rescheduled, within given limits, from the already dispatched levels obtained from the energy market clearing process. The proposed dispatch model achieves the main objective of an ISO in a competitive electricity market, which is to provide the required reactive power support from generators at least cost while ensuring a secure operation of the power system. The proposed reactive power procurement and dispatch models capture both the technical and economic aspects of power system operation in competitive electricity markets; however, from an optimization point of view, these models represent non-convex mixed integer non-linear programming (MINLP) problems due to the presence of binary variables associated with the different regions of reactive power operation in a synchronous generator. Such MINLP optimization problems are difficult to solve, especially for an actual power system. A novel Generator Reactive Power Classification (GRPC) algorithm is proposed in this thesis to address this issue, with the advantage of iteratively solving the optimization models as a series of non-linear programming (NLP) sub-problems. The proposed reactive power procurement and dispatch models are implemented and tested on the CIGRE 32-bus system, with several case studies that represent different practical operating scenarios. The developed models are also compared with other approaches for reactive power provision, and the results demonstrate the robustness and effectiveness of the proposed model. The results clearly reveal the main features of the proposed models for optimal provision of reactive power ancillary service, in order to suit the requirements of

  4. Probing the functional subunits of the tonoplast H+-ATPase

    International Nuclear Information System (INIS)

    Randall, S.K.; Lai, S.; Sze, H.

    1986-01-01

    The tonoplast ATPase of oat roots is composed of at least three polypeptides of 72, 60, and 16 kDa. The 16 kDA polypeptide covalently binds N,N'-dicyclohexylcarbodiimide and is postulated to be a component of the proton channel. Initial studies to identify other subunits indicate that both the 72 and 60 kDa subunits covalently bind 14 C]-7-chloro-4-nitrobenzo-2-oxa-1,3-diazole and [ 14 C]N-ethylamleimide, inhibitors of the tonoplast ATPase. ATP prevents binding of these inhibitors suggesting that both the 72 and 60 kDa subunits are involved in substrate binding. Polyclonal antibody has been made to the 72 kDa subunit. Western blot analysis of tonoplast vesicles reveals single reactive polypeptide (72 kDa). The antibody shows no cross-reactivity towards either the mitochondrial F 1 -ATPase or the plasma membrane ATPase. This antibody specifically inhibits ATP hydrolysis and ATP-dependent H + pumping in native tonoplast vesicles. The authors conclude that the 72 kDa subunit is intimately associated with the catalytic (or ATP-binding) site

  5. Separation and characterization of alpha-chain subunits from tilapia (Tilapia zillii) skin gelatin using ultrafiltration.

    Science.gov (United States)

    Chen, Shulin; Tang, Lanlan; Su, Wenjin; Weng, Wuyin; Osako, Kazufumi; Tanaka, Munehiko

    2015-12-01

    Alpha-chain subunits were separated from tilapia skin gelatin using ultrafiltration, and the physicochemical properties of obtained subunits were investigated. As a result, α1-subunit and α2-subunit could be successfully separated by 100 kDa MWCO regenerated cellulose membranes and 150 kDa MWCO polyethersulfone membranes, respectively. Glycine was the most dominant amino acid in both α1-subunit and α2-subunit. However, the tyrosine content was higher in α2-subunit than in α1-subunit, resulting in strong absorption near 280 nm observed in the UV absorption spectrum. Based on the DSC analysis, it was found that the glass transition temperatures of gelatin, α1-subunit and α2-subunit were 136.48 °C, 126.77 °C and 119.43 °C, respectively. Moreover, the reduced viscosity and denaturation temperature of α1-subunit were higher than those of α2-subunit, and the reduced viscosity reached the highest when α-subunits were mixed with α1/α2 ratio of approximately 2, suggesting that α1-subunit plays a more important role in the thermostability of gelatin than α2-subunit. Copyright © 2015 Elsevier Ltd. All rights reserved.

  6. Dynamic properties of motor proteins with two subunits

    International Nuclear Information System (INIS)

    Kolomeisky, Anatoly B; III, Hubert Phillips

    2005-01-01

    The dynamics of motor protein molecules consisting of two subunits is investigated using simple discrete stochastic models. Exact steady-state analytical expressions are obtained for velocities and dispersions for any number of intermediate states and conformations between the corresponding binding states of proteins. These models enable us to provide a detailed description and comparison of two different mechanisms of the motion of motor proteins along the linear tracks: the hand-over-hand mechanism, when the motion of subunits alternate; and the inchworm mechanism, when one subunit is always trailing another one. It is shown that the proteins in the hand-over-hand mechanism move faster and fluctuate more than the molecules in the inchworm mechanism. The effect of external forces on dynamic properties of motor proteins is also discussed. Finally, a quantitative method, based on experimental observations for single motor proteins, is proposed for distinguishing between two mechanisms of motion

  7. Novel subunit structure observed for noncooperative hemoglobin from Urechis caupo.

    Science.gov (United States)

    Kolatkar, P R; Meador, W E; Stanfield, R L; Hackert, M L

    1988-03-05

    Tetrameric hemoglobin from the "fat innkeeper" worm Urechis caupo possesses a novel subunit arrangement having an "inside out" quaternary structure in that the G/H helices are located on the outer surface of the tetramer. A 5-A resolution crystal structure reveals that although the individual subunits are beta-like, having a distinct D helix and the general myoglobin fold, the subunit contacts are very different from those previously observed for hemoglobins. Furthermore, the hemoglobin from U. caupo is also quite different from the unusual hemoglobin tetramer from clam which also has its G/H helices on the outer surface but with the hemes in close proximity through E-F helical contacts (Royer, W. E., Jr., Love, W. E., and Fenderson, F. F. (1985) Nature 316, 277-280).

  8. CSNAP Is a Stoichiometric Subunit of the COP9 Signalosome

    Directory of Open Access Journals (Sweden)

    Shelly Rozen

    2015-10-01

    Full Text Available The highly conserved COP9 signalosome (CSN complex is a key regulator of all cullin-RING-ubiquitin ligases (CRLs, the largest family of E3 ubiquitin ligases. Until now, it was accepted that the CSN is composed of eight canonical components. Here, we report the discovery of an additional integral and stoichiometric subunit that had thus far evaded detection, and we named it CSNAP (CSN acidic protein. We show that CSNAP binds CSN3, CSN5, and CSN6, and its incorporation into the CSN complex is mediated through the C-terminal region involving conserved aromatic residues. Moreover, depletion of this small protein leads to reduced proliferation and a flattened and enlarged morphology. Finally, on the basis of sequence and structural properties shared by both CSNAP and DSS1, a component of the related 19S lid proteasome complex, we propose that CSNAP, the ninth CSN subunit, is the missing paralogous subunit of DSS1.

  9. Cholera Toxin B: One Subunit with Many Pharmaceutical Applications

    Directory of Open Access Journals (Sweden)

    Keegan J. Baldauf

    2015-03-01

    Full Text Available Cholera, a waterborne acute diarrheal disease caused by Vibrio cholerae, remains prevalent in underdeveloped countries and is a serious health threat to those living in unsanitary conditions. The major virulence factor is cholera toxin (CT, which consists of two subunits: the A subunit (CTA and the B subunit (CTB. CTB is a 55 kD homopentameric, non-toxic protein binding to the GM1 ganglioside on mammalian cells with high affinity. Currently, recombinantly produced CTB is used as a component of an internationally licensed oral cholera vaccine, as the protein induces potent humoral immunity that can neutralize CT in the gut. Additionally, recent studies have revealed that CTB administration leads to the induction of anti-inflammatory mechanisms in vivo. This review will cover the potential of CTB as an immunomodulatory and anti-inflammatory agent. We will also summarize various recombinant expression systems available for recombinant CTB bioproduction.

  10. Dengue vaccine: an update on recombinant subunit strategies.

    Science.gov (United States)

    Martin, J; Hermida, L

    2016-03-01

    Dengue is an increasing public health problem worldwide, with the four serotypes of the virus infecting over 390 million people annually. There is no specific treatment or antiviral drug for dengue, and prevention is largely limited to controlling the mosquito vectors or disrupting the human-vector contact. Despite the considerable progress made in recent years, an effective vaccine against the virus is not yet available. The development of a dengue vaccine has been hampered by many unique challenges, including the need to ensure the absence of vaccine-induced enhanced severity of disease. Recombinant protein subunit vaccines offer a safer alternative to other vaccine approaches. Several subunit vaccine candidates are presently under development, based on different structural and non-structural proteins of the virus. Novel adjuvants or immunopotentiating strategies are also being tested to improve their immunogenicity. This review summarizes the current status and development trends of subunit dengue vaccines.

  11. Isolation and properties of a succinylated subunit of human thyroglobulin.

    Science.gov (United States)

    Smith, D J; Shulman, S

    1978-03-01

    A subunit of succinylated (40:1 molar ratio, succinic anhydride:lysine residues) human thyroglobulin (Tg) was prepared by gel filtration on 6% and 4% agarose. This subunit (S-8) had a sdegrees20, w of 7.6S, Ddegrees20, w of 2.96 x 10(7) and an equilibrium molecular weight of 165,000 after correction for bound succinic anhydride (SA). The S-8 component was suggested to be a quarter unit of intact Tg. The S-8 and intact Tg had nearly identical amino acid compositions with differences only in glutamic acid, glycine and proline. The iodine content of both components was similar. The succinylated S-8 subunit, as well as material which sedimented with a value of 2S or less, retained some but not all heteroantigenic and autoantigenic sites in immunodiffusion and inhibition of passive hemagglutination.

  12. Partial agonists and subunit selectivity at NMDA receptors

    DEFF Research Database (Denmark)

    Risgaard, Rune; Hansen, Kasper Bø; Clausen, Rasmus Prætorius

    2010-01-01

    -methyl-D-aspartic acid (NMDA) receptor class. Development of these ligands seems to be a difficult task because of the conserved region in the binding site of the NMDA receptor subunits. A few scaffolds have been developed showing potential to differentiate between the NMDA receptors.......Subunit-selective ligands for glutamate receptors remains an area of interest as glutamate is the major excitatory neurotransmitter in the brain and involved in a number of diseased states in the central nervous system (CNS). Few subtype-selective ligands are known, especially among the N...

  13. Engineering evaluation/cost analysis for the 105-DR and 105-F Reactor facilities and ancillary facilities

    International Nuclear Information System (INIS)

    Coenenberg, E.T.

    1998-01-01

    This document presents the results of an engineering evaluation/cost analysis (EE/CA) that was conducted to evaluate alternatives to address final disposition of the 105-DR and 105-F Reactor Buildings (subsequently referred to as facilities), including the fuel storage basins (FSB) and below-grade portions of the reactors, excluding the reactor blocks. The reactor blocks will remain in a safe storage mode for up to 75 years as identified in the Record the Decision (ROD) (58 FR 48509) for the Environmental Impact Statement (EIS), Decommissioning of Eight Surplus Production Reactors at the Hanford Site, Richland, Washington (DOE 1992a). This EE/CA also addresses final disposition of four ancillary facilities: 116-D and 116-DR Exhaust Air Stacks, 117-DR Exhaust Filter Building, and 119-DR Exhaust Air Sample Building. The 105-DR and 105-F facilities are located in the 100-D and 100-F Areas of the Hanford Site. In November 1989, the 100 Area of the Hanford Site was placed on the U.S. Environmental Protection Agency's (EPA) National Priorities List (NPL) under the Comprehensive Environmental Response, Compensation, and Liability Act of 1980 (CERCLA). The 100 Area NPL includes the 100-D Area (which includes the 100-DR site) and the 100-F Area, which are in various stages of the remediation process. It has been determined by DOE that hazardous substances in the 105-DR, 105-F, and the four ancillary facilities may present a potential threat to human health or the environment, and that a non-time critical removal action at these facilities is warranted. To help determine the most appropriate action, DOE, in cooperation with the Washington State Department of Ecology (Ecology) and the EPA, has prepared this EE/CA. The scope of the evaluation includes the 105-DR and 105-F facilities and the four ancillary facilities. The 116-DR and 117-DR facilities are located within the boundaries of the 105-DR Large Sodium Fire Facility Treatment, Storage, and Disposal (TSD) unit, which is

  14. SU-F-I-72: Evaluation of the Ancillary Lead Shielding for Optimizing Radiation Protection in the Interventional Radiology Department

    Energy Technology Data Exchange (ETDEWEB)

    Tonkopi, E; Lightfoot, C [Dalhousie University, Queen Elizabeth II Health Sciences Ctr, Halifax, NS (Canada); LeBlanc, E [Queen Elizabeth II Health Sciences Ctr, Halifax, NS (Canada)

    2016-06-15

    Purpose: The rising complexity of interventional fluoroscopic procedures has resulted in an increase of occupational radiation exposures in the interventional radiology (IR) department. This study assessed the impact of ancillary shielding on optimizing radiation protection for the IR staff. Methods: Scattered radiation measurements were performed in two IR suites equipped with Axiom Artis systems (Siemens Healthcare, Erlangen, Germany) installed in 2006 and 2010. Both rooms had suspended ceiling-mounted lead-acrylic shields of 75×60 cm (Mavig, Munich, Germany) with lead equivalency of 0.5 mm, and under-table drapes of 70×116 cm and 65×70 cm in the newer and the older room respectively. The larger skirt can be wrapped around the table’s corner and in addition the newer suite had two upper shields of 25×55 cm and 25×35 cm. The patient was simulated by 30 cm of acrylic, air kerma rate (AKR) was measured with the 180cc ionization chamber (AccuPro Radcal Corporation, Monrovia, CA, USA) at different positions. The ancillary shields, x-ray tube, image detector, and table height were adjusted by the IR radiologist to simulate various clinical setups. The same exposure parameters were used for all acquisitions. AKR measurements were made at different positions relative to the operator. Results: The AKR measurements demonstrated 91–99% x-ray attenuation by the drapes in both suites. The smaller size of the under-table skirt and absence of the side-drapes in the older room resulted in a 20–50 fold increase of scattered radiation to the operator. The mobile suspended lead-acrylic shield reduced AKR by 90–94% measured at 150–170 cm height. The recommendations were made to replace the smaller under-table skirt and to use the ceiling-mounted shields for all IR procedures. Conclusion: The ancillary shielding may significantly affect radiation exposure to the IR staff. The use of suspended ceiling-mounted shields is especially important for reduction of

  15. Finisher hog production in the Southeastern United States: Ancillary measurements derived from the National Air Emissions Monitoring Study (NAEMS)

    Science.gov (United States)

    Robarge, W. P.; Lee, S.; Walker, J. T.

    2010-12-01

    Measurements of emissions of gases and fine particulate matter from swine animal feeding operations (AFOs) in the southeastern US have typically been confined to relatively short periods (days to several weeks) and have generally focused on waste lagoons. Access to swine animal housing units and other ancillary information has been limited. The National Air Emissions Monitoring Study (NAEMS) provided a unique opportunity to characterize emissions from swine housing units for an extended period of time (~ 2 years), and allowed access to ancillary measurements regarding nutrient flows (feed amounts and composition), manure dynamics, animal inventories, water usage and farm management. Presented here is a summary of the observations made for a NAEMS finisher site (NC3B) selected as being representative of swine production in the southeastern US. Finisher hogs are raised in rotations (~ 140 days) with a target market weight of 123 kg/hog. Among the population during a rotation (700-800 hogs/barn) the actual growth rate varies with a series of “grade-outs” of market-weight hogs starting ~ 110 days from initial load-in. Derivation of the standing live-weight in the barns during a rotation therefore requires use of a growth model and summation over several different “populations” of hogs within a single barn. Up to 5 different feed formulations are fed during a rotation with %N content ranging from (3.4 to 2.2% N; total feed consumed 181,000 kg/barn). Across 4 complete rotations, N consumed was ~50 g N per hog/day. Of this amount, we estimate ~ 60% is excreted as fecal matter and urine. The TAN (NH3 + NH4+) content of the shallow pits is consistently higher (1880 ±390 mg TAN/L) than that found in the anaerobic lagoon (800 ±70 mg TAN/L), except immediately after recharge following pit-pull (pH of the two liquids was similar). The presence of a recalcitrant layer of sludge in the shallow pits (liquid height = 20 cm; sludge depth = 5-10 cm; TAN = 2500 mg N/L; total

  16. C25, an essential RNA polymerase III subunit related to the RNA polymerase II subunit RPB7.

    OpenAIRE

    Sadhale, P P; Woychik, N A

    1994-01-01

    We identified a partially sequenced Saccharomyces cerevisiae gene which encodes a protein related to the S. cerevisiae RNA polymerase II subunit, RPB7. Several lines of evidence suggest that this related gene, YKL1, encodes the RNA polymerase III subunit C25. C25, like RPB7, is present in submolar ratios, easily dissociates from the enzyme, is essential for cell growth and viability, but is not required in certain transcription assays in vitro. YKL1 has ABF-1 and PAC upstream sequences often ...

  17. Subunit compensation and plasticity of synaptic GABAA receptors induced by ethanol in α4 subunit knockout mice

    Directory of Open Access Journals (Sweden)

    Asha eSuryanarayanan

    2011-09-01

    Full Text Available There is considerable evidence that ethanol (EtOH potentiates γ-aminobutyric acid type A receptor (GABAAR action, but only GABAARs containing δ subunits appear sensitive to low mM EtOH. The α4 and δ subunits co-assemble into GABAARs which are relatively highly expressed at extrasynaptic locations in the dentate gyrus where they mediate tonic inhibition. We previously demonstrated reversible- and time-dependent changes in GABAAR function and subunit composition in rats after single-dose EtOH intoxication. We concluded that early tolerance to EtOH occurs by over-activation and subsequent internalization of EtOH-sensitive extrasynaptic α4βδ-GABAARs. Based on this hypothesis, any highly EtOH-sensitive GABAARs should be subject to internalization following exposure to suitably high EtOH doses. To test this, we studied the GABAARs in mice with a global deletion of the α4 subunit (KO. The dentate granule cells (DGCs of these mice exhibited greatly reduced tonic currents and greatly reduced potentiation by acutely applied EtOH, whereas synaptic currents showed heightened sensitivity to low EtOH concentrations. The hippocampus of naive KO mice showed reduced δ subunit protein levels, but increased α2, and γ2 levels compared to wild-type (WT controls, suggesting at least partial compensation by these subunits in synaptic, highly EtOH-sensitive GABAARs of KO mice. In WT mice, cross-linking and Western blot analysis at 1 h after an EtOH challenge (3.5 g/kg, i.p. revealed increased intracellular fraction of the α1, α4 and δ, but not α2, α5 or γ2 subunits. By contrast, we observed significant internalization of α1, α2, δ, and γ2 subunits after a similar EtOH challenge in KO mice. Synaptic currents from naïve KO mice were more sensitive to potentiation by zolpidem (0.3 μM, requiring α1/α2, inactive at α4/5 GABAARs than those from naïve WT mice. At 1 h after EtOH, synaptic currents of WT mice were unchanged, whereas those of KO mice

  18. Methodology and forecast products for the optimal offering of ancillary services from wind in a market environment

    DEFF Research Database (Denmark)

    Bacher, Peder; Madsen, Henrik; Pinson, Pierre

    of wind power for example in the planning of ancillary power services, where the level of available wind power with a high degree of certainty is important to know. The presented extreme value models are applied to negative forecast residuals from state-of-the-art wind power forecast software...... of extreme weather induced phenomena, for example extreme water levels in a river, wind levels or at sea for design of dykes (de Haan and de Ronde, 1998). In insurance and finance the extreme value modelling is widespread (Embrechts et al., 1997). Extreme value statistics for energy and power applications...... is also widely used, for example for planning in wind power operation (Horvat et al., 2013) and peak wind prediction (Cook, 1982) and (Friederichs and Thorarinsdottir, 2012). Several books provide comprehensive introductions to extreme value theory, for example Coles (2001) and Beirlant et al. (2006...

  19. Climate policy and ancillary benefits. A survey and integration into the modelling of international negotiations on climate change

    International Nuclear Information System (INIS)

    Pittel, Karen; Ruebbelke, Dirk T.G.

    2008-01-01

    Currently informal and formal international negotiations on climate change take place in an intensive way since the Kyoto Protocol expires already in 2012. A post-Kyoto regulation to combat global warming is not yet stipulated. Due to rapidly increasing greenhouse-gas emission levels, industrialized countries urge major polluters from the developing world like China and India to participate in a future agreement. Whether these developing countries will do so, depends on the prevailing incentives to participate in international climate protection efforts. This paper identifies ancillary benefits of climate policy to provide important incentives to attend a new international protocol and to positively affect the likelihood of accomplishing a post-Kyoto agreement which includes commitments of developing countries. (author)

  20. Risk-constrained dynamic self-scheduling of a pumped-storage plant in the energy and ancillary service markets

    International Nuclear Information System (INIS)

    Kazempour, S. Jalal; Moghaddam, M. Parsa; Haghifam, M.R.; Yousefi, G.R.

    2009-01-01

    This work addresses a new framework for self-scheduling of an individual price-taker pumped-storage plant in a day-ahead (DA) market. The goal is achieving the best trade-off between the expected profit and the risks when the plant participates in DA energy, spinning reserve and regulation markets. In this paper, a set of uncertainties including price forecasting errors and also the uncertainty of power delivery requests in the ancillary service markets are contemplated. Considering these uncertainties, a new approach is proposed which is called dynamic self-scheduling (DSS). This risk-constrained dynamic self-scheduling problem is therefore formulated and solved as a mixed integer programming (MIP) problem. Numerical results for a case study are discussed. (author)

  1. α-4 subunit of nicotinic acetylcholine receptor polymorphisms exhibit ...

    African Journals Online (AJOL)

    Background: Smoking behavior is influenced by both genetic and environmental factors. Nicotine is the major addictive substance in cigarettes. Nicotinic acetylcholine receptors (nAChRs) are thought to play an important role in nicotine addiction of smokers. One of the genes, α-4 subunit of nicotinic acetylcholine receptor ...

  2. Thermostable Subunit Vaccines for Pulmonary Delivery: How Close Are We?

    DEFF Research Database (Denmark)

    Foged, Camilla

    2016-01-01

    , such as influenza, tuberculosis, and Ebola, for which no good universal vaccines exist. At least two pharmaceutical improvements are expected to help filling this gap: i) The development of thermostable vaccine dosage forms, and ii) the full exploitation of the adjuvant technology for subunit vaccines to potentiate...

  3. Allelic variation of HMW glutenin subunits of Ethiopian bread wheat ...

    African Journals Online (AJOL)

    There were highly significant differences between genotypes and banding patterns for the SDS-sedimentation test, mixograph development time, alveograph strength and loaf volume; but not for protein content. The frequency of subunits 5+10 among genotypes was 73%. The accumulation of high scoring alleles in our ...

  4. Editing modifies the GABA(A) receptor subunit alpha3

    DEFF Research Database (Denmark)

    Ohlson, Johan; Pedersen, Jakob Skou; Haussler, David

    2007-01-01

    to find selectively edited sites and combined it with bioinformatic techniques that find stem-loop structures suitable for editing. We present here the first verified editing candidate detected by this screening procedure. We show that Gabra-3, which codes for the alpha3 subunit of the GABA(A) receptor...

  5. Development of a Subunit Vaccine for Contagious Bovine ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Their work has set the stage for commercial development of a sub-unit vaccine. Field testing and production This scale-up project will ... The research team will conduct a large field trial involving 1,500 commercial cattle to assess the vaccine's safety and efficacy. They will define the regulatory pathway for the vaccine to be ...

  6. weight glutenin subunits and waxy alleles on dough-mix

    Indian Academy of Sciences (India)

    weight glutenin subunits and waxy alleles on dough-mixing properties in common wheat. Zhiying Deng, Shuna Hu, Feifei Zheng, Junnan Chen, Xinye Zhang, Jiansheng Chen, Cailing Sun,. Yongxiang Zhang, Shouyi Wang and Jichun Tian. J. Genet. 92, 69–79. Table 1. The data of the mixing properties of the RIL population ...

  7. Bacterial cellulose biosynthesis: diversity of operons, subunits, products and functions

    Science.gov (United States)

    Römling, Ute; Galperin, Michael Y.

    2015-01-01

    Summary Recent studies of bacterial cellulose biosynthesis, including structural characterization of a functional cellulose synthase complex, provided the first mechanistic insight into this fascinating process. In most studied bacteria, just two subunits, BcsA and BcsB, are necessary and sufficient for the formation of the polysaccharide chain in vitro. Other subunits – which differ among various taxa – affect the enzymatic activity and product yield in vivo by modulating expression of biosynthesis apparatus, export of the nascent β-D-glucan polymer to the cell surface, and the organization of cellulose fibers into a higher-order structure. These auxiliary subunits play key roles in determining the quantity and structure of the resulting biofilm, which is particularly important for interactions of bacteria with higher organisms that lead to rhizosphere colonization and modulate virulence of cellulose-producing bacterial pathogens inside and outside of host cells. Here we review the organization of four principal types of cellulose synthase operons found in various bacterial genomes, identify additional bcs genes that encode likely components of the cellulose biosynthesis and secretion machinery, and propose a unified nomenclature for these genes and subunits. We also discuss the role of cellulose as a key component of biofilms formed by a variety of free-living and pathogenic bacteria and, for the latter, in the choice between acute infection and persistence in the host. PMID:26077867

  8. Evaluation of subunit vaccines against feline immunodeficiency virus infection

    NARCIS (Netherlands)

    Horzinek, M.C.; Verschoor, E.J.; Willemse, M.J.; Stam, J.G.; Vliet, A.L.W. van; Pouwels, H.; Chalmers, S.K.; Sondermeijer, P.J.; Hesselink, W.; Ronde, A. de

    1996-01-01

    Subunit vaccines prepared against feline immunodeficiency virus (FIV) infection were evaluated in two trials. First, cats were immunized with bacterial expression products of an envelope fragment that contained the V3 neutralization domain of the FIV surface protein fused to either galactokinase

  9. Characterization of low-molecular-weight glutenin subunit genes of ...

    Indian Academy of Sciences (India)

    2015-09-16

    Sep 16, 2015 ... species. Materials and methods. Plant materials. A total of 13 accessions from five species of Sitopsis were used for gene cloning (table 1 in electronic supplementary. Keywords. wheat; Aegilops; low-molecular-weight glutenin subunits; gene cloning; B genome; Triticum aestivum. Journal of Genetics, Vol.

  10. Purification of subunits from human chorionic gonadotropin and radioimmunoassay

    International Nuclear Information System (INIS)

    Okumura, Hajime; Sudo, Tadamitsu; Fujisawa, Munetoshi; Namba, Shuichi; Matsushima, Sanae

    1976-01-01

    A crude hCG with an activity of about 3,000 IU per mg was purified to 10,000-15,000 IU per mg of dry weight using Amberlite CG-50 chromatography combined with DEAE-Sephadex A-50 and Sephadex G-75. The alpha and beta subunits of hCG were prepared by urea-treatment of the hormone and were isolated by DEAE-Sephadex A-50 chromatography. Further purification of the subunits was achieved by gel filtration on a Sephadex G-57 column. For radioimmunoassay, hCG was iodinated by the DMSO-chloramine T method. Iodination of hCG with non-radioactive iodine revealed that the addition of DMSO to the iodination mixture seemed to reduce the iodination damage to the antigenic activity of the hormone. The hCG iodinated with non-radioactive iodine (accomplished by the DMSO-chloramine T method) was 1.5 times more immunoreactive in the hCG radioimmunoassay then the hCG iodinated by the usual chloramine T method. The radioimmunoassay of the hCG-beta subunit developed in our laboratory was satisfactory with respect to specificity. The hLH, hFSH, hTSH and hCG-alpha subunits tested cross-reacted very poorly in our assay system. Desialylated-hCG and subunits, in which the biologic potency was almost zero, also exhibited decreased immunoreactivity, about 30% of the native hormone with grossly unimpaired parallelism in their respective homologous radioimmunoassays. The concentrations of hCG and the subunits were determined on human sera from pregnant patients. The hCG levels reached to a peak at the first trimester of the pregnancy, but the hCG-beta subunits varied poorly in their concentrations throughout the periods of pregnancy. The hCG-alpha levels, on the other hand, showed two distinct peaks, in the early period and at the term of pregnancy. (J.P.N.)

  11. Different effort constructs and effort-reward imbalance: effects on employee well-being in ancillary health care workers.

    Science.gov (United States)

    van Vegchel, N; de Jonge, J; Meijer, T; Hamers, J P

    2001-04-01

    The present study investigates the relationship between Effort-Reward Imbalance (ERI) and employee well-being, using three different concepts of efforts (i.e. psychological demands, physical demands and emotional demands). The ERI model had been used as a theoretical framework, indicating that work stress is related to high efforts (i.e. job demands) and low occupational rewards (e.g. money, esteem and security/career opportunities). The ERI model also predicts that, in overcommitted workers, effects of ERI on employee well-being are stronger compared with their less committed counterparts. A cross-sectional survey among 167 ancillary health care workers of two nursing homes was conducted. Multiple univariate logistic regression analyses were used to test the relationship between ERI and employee well-being. Results of the logistic regression analyses showed that employees with both high (psychological, physical and emotional) efforts and low rewards had higher risks of psychosomatic health complaints, physical health symptoms and job dissatisfaction (odds ratios (ORs) ranged from 5.09 to 18.55). Moreover, employees who reported both high efforts and high rewards had elevated risks of physical symptoms and exhaustion (ORs ranged from 6.17 to 9.39). No support was found for the hypothesis on the moderating effect of overcommitment. Results show some support for the ERI model; ancillary health care workers with high effort/low reward imbalance had elevated risks of poor employee well-being. In addition, results show that the combination of high efforts and high rewards is important for employee well-being. Finally, some practical implications are discussed to combat work stress in health care work.

  12. Development of an Asset Value Map for Disaster Risk Assessment in China by Spatial Disaggregation Using Ancillary Remote Sensing Data.

    Science.gov (United States)

    Wu, Jidong; Li, Ying; Li, Ning; Shi, Peijun

    2018-01-01

    The extent of economic losses due to a natural hazard and disaster depends largely on the spatial distribution of asset values in relation to the hazard intensity distribution within the affected area. Given that statistical data on asset value are collected by administrative units in China, generating spatially explicit asset exposure maps remains a key challenge for rapid postdisaster economic loss assessment. The goal of this study is to introduce a top-down (or downscaling) approach to disaggregate administrative-unit level asset value to grid-cell level. To do so, finding the highly correlated "surrogate" indicators is the key. A combination of three data sets-nighttime light grid, LandScan population grid, and road density grid, is used as ancillary asset density distribution information for spatializing the asset value. As a result, a high spatial resolution asset value map of China for 2015 is generated. The spatial data set contains aggregated economic value at risk at 30 arc-second spatial resolution. Accuracy of the spatial disaggregation reflects redistribution errors introduced by the disaggregation process as well as errors from the original ancillary data sets. The overall accuracy of the results proves to be promising. The example of using the developed disaggregated asset value map in exposure assessment of watersheds demonstrates that the data set offers immense analytical flexibility for overlay analysis according to the hazard extent. This product will help current efforts to analyze spatial characteristics of exposure and to uncover the contributions of both physical and social drivers of natural hazard and disaster across space and time. © 2017 Society for Risk Analysis.

  13. The role of liquid-based cytology and ancillary techniques in pleural and pericardic effusions: an institutional experience.

    Science.gov (United States)

    Rossi, Esther Diana; Bizzarro, Tommaso; Schmitt, Fernando; Longatto-Filho, Adhemar

    2015-04-01

    Fine-needle aspiration cytology (FNAC) of serous membrane effusions may fulfil a challenging role in the diagnostic analysis of both primary and metastatic disease. From this perspective, liquid-based cytology (LBC) represents a feasible and reliable method for empowering the performance of ancillary techniques (ie, immunocytochemistry and molecular testing) with high diagnostic accuracy. In total, 3171 LBC pleural and pericardic effusions were appraised between January 2000 and December 2013. They were classified as negative for malignancy (NM), suspicious for malignancy (SM), or positive for malignancy (PM). The cytologic diagnoses included 2721 NM effusions (2505 pleural and 216 pericardic), 104 SM effusions (93 pleural and 11 pericardic), and 346 PM effusions (321 pleural and 25 pericardic). The malignant pleural series included 76 unknown malignancies (36 SM and 40 PM effusions), 174 metastatic lesions (85 SM and 89 PM effusions), 14 lymphomas (3 SM and 11 PM effusions), 16 mesotheliomas (5 SM and 11 SM effusions), and 3 myelomas (all SM effusions). The malignant pericardic category included 20 unknown malignancies (5 SM and 15 PM effusions), 15 metastatic lesions (1 SM and 14 PM effusions), and 1 lymphoma (1 PM effusion). There were 411 conclusive immunocytochemical analyses and 47 molecular analyses, and the authors documented 88% sensitivity, 100% specificity, 98% diagnostic accuracy, 98% negative predictive value, and 100% positive predictive value for FNAC. FNAC represents a primary diagnostic tool for effusions and a reliable approach with which to determine the correct follow-up. Furthermore, LBC is useful for ancillary techniques, such as immunocytochemistry and molecular analysis, with feasible diagnostic and predictive utility. © 2015 American Cancer Society.

  14. The Role of Liquid Based Cytology and Ancillary Techniques in the Peritoneal Washing Analysis: Our Institutional Experience

    Science.gov (United States)

    Rossi, Esther; Bizzarro, Tommaso; Martini, Maurizio; Longatto-Filho, Adhemar; Schmitt, Fernando; Fagotti, Anna; Scambia, Giovanni; Zannoni, Gian Franco

    2017-01-01

    Background The cytological analysis of peritoneal effusions serves as a diagnostic and prognostic aid for either primary or metastatic diseases. Among the different cytological preparations, liquid based cytology (LBC) represents a feasible and reliable method ensuring also the application of ancillary techniques (i.e immunocytochemistry-ICC and molecular testing). Methods We recorded 10348 LBC peritoneal effusions between January 2000 and December 2014. They were classified as non-diagnostic (ND), negative for malignancy-NM, atypical-suspicious for malignancy-SM and positive for malignancy-PM. Results The cytological diagnosis included 218 ND, 9.035 NM, 213 SM and 882 PM. A total of 8048 (7228 NM, 115SM, 705 PM) cases with histological follow-up were included. Our NM included 21 malignant and 7207 benign histological diagnoses. Our 820 SMs+PMs were diagnosed as 107 unknown malignancies (30SM and 77PM), 691 metastatic lesions (81SM and 610PM), 9 lymphomas (2SM and 7PM), 9 mesotheliomas (1SM and 8SM), 4 sarcomas (1SM and 3PM). Primary gynecological cancers contributed with 64% of the cases. We documented 97.4% sensitivity, 99.9% specificity, 98% diagnostic accuracy, 99.7% negative predictive value (NPV) and 99.7% positive predictive value (PPV). Furthermore, the morphological diagnoses were supported by either 173 conclusive ICC results or 50 molecular analyses. Specifically the molecular testing was performed for the EGFR and KRAS mutational analysis based on the previous or contemporary diagnoses of Non Small Cell Lung Cancer (NSCLC) and colon carcinomas. We identified 10 EGFR in NSCCL and 7 KRAS mutations on LBC stored material. Conclusions Peritoneal cytology is an adjunctive tool in the surgical management of tumors mostly gynecological cancers. LBC maximizes the application of ancillary techniques such as ICC and molecular analysis with feasible diagnostic and predictive yields also in controversial cases. PMID:28099523

  15. Novel phosphorescent cationic iridium(iii) complexes with o-carboranylation on the ancillary N^N ligand.

    Science.gov (United States)

    Li, Xiang; Yin, Yongheng; Yan, Hong; Lu, Changsheng; Zhao, Qiang

    2017-08-14

    A novel series of heteroleptic iridium complexes with 2-phenyl-pyridine as a main ligand and carborane-functionalized 2,2'-bipyridine as an ancillary ligand were synthesized, and characterized as [Ir(ppy) 2 (By)]PF 6 (where ppy is 2-phenyl-pyridine, By is 5-(2-R-Cb)-2,2'-bipyridine, R = H (2a), CH 3 (2b), Ph (2c), iPr (2d) and iBu (2e), or By is 4-(2-R-Cb)-2,2'-bipyridine while R = H (3a), CH 3 (3b), Ph (3c), iPr (3d) and iBu (3e), Cb = o-carboran-1-yl). The R groups and the substitution sites of carborane on the pyridine ring have caused differences in the emission properties of these complexes. In addition, the quantum efficiency of [Ir(ppy) 2 (By)]PF 6 complexes has been tuned as well through the introduction of various 2-R-substituted o-carboranes into the ancillary ligand 2,2'-bipyridine, no matter in the solid state (from 0.12 to 0.25) or in solution (from 0.04 to 0.25). The emission color was tuned from yellow to red by the o-carboranyl unit because of its inductive effect. Density functional theory (DFT) and time dependent DFT (TD-DFT) calculations have been applied to investigate excited-state electronic structures of the newly synthesized complexes, which are consistent with the observed red-shift emissions.

  16. Improvements in the in-beam γ-ray spectroscopy provided by an ancillary detector coupled to a Ge γ-spectrometer: the DIAMANT-EUROGAM II example

    Science.gov (United States)

    Scheurer, J. N.; Aiche, M.; Aleonard, M. M.; Barreau, G.; Bourgine, F.; Boivin, D.; Cabaussel, D.; Chemin, J. F.; Doan, T. P.; Goudour, J. P.; Harston, M.; Brondi, A.; La Rana, G.; Moro, R.; Vardaci, E.; Curien, D.

    1997-02-01

    For the first time the 4π γ-ray spectrometer EUROGAM II has been coupled to a 4π light charged particle detector array, DIAMANT, during a test experiment on the reaction 32S + 58Ni at 120 MeV beam energy. A very large improvement in the peak-to-background ratio of the γ-spectra has been found when EUROGAM II is triggered by DIAMANT to select an exit channel. A simple algebra has been developed which provides theoretical estimates in good agreement with these experimental results. It is demonstrated that, depending on both the γ-spectrometer and ancillary detector performances, much better peak-to-background can be obtained by such a coupling. For the same peak-to-background ratio, the use of an ancillary detector allows for a lower γ-ray coincidence level and therefore improves the statistics. Ways to select the most appropriate ancillary detector are given. The ability of the ancillary detector to provide a total Doppler shift correction is crucial for the improvement of the peak-to-background ratio.

  17. Prediction of spatial soil property information from ancillary sensor data using ordinary linear regression: Model derivations, residual assumptions and model validation tests

    Science.gov (United States)

    Geospatial measurements of ancillary sensor data, such as bulk soil electrical conductivity or remotely sensed imagery data, are commonly used to characterize spatial variation in soil or crop properties. Geostatistical techniques like kriging with external drift or regression kriging are often use...

  18. Effect of high and low molecular weight glutenin subunits, and subunits of gliadin on physicochemical parameters of different wheat genotypes

    Directory of Open Access Journals (Sweden)

    Mariana Souza Costa

    2013-02-01

    Full Text Available Identification of functional properties of wheat flour by specific tests allows genotypes with appropriate characteristics to be selected for specific industrial uses. The objective of wheat breeding programs is to improve the quality of germplasm bank in order to be able to develop wheat with suitable gluten strength and extensibility for bread making. The aim of this study was to evaluate 16 wheat genotypes by correlating both glutenin subunits of high and low molecular weight and gliadin subunits with the physicochemical characteristics of the grain. Protein content, sedimentation volume, sedimentation index, and falling number values were analyzed after the grains were milled. Hectoliter weight and mass of 1000 seeds were also determined. The glutenin and gliadin subunits were separated using polyacrylamide gel in the presence of sodium dodecyl sulfate. The data were evaluated using variance analysis, Pearson's correlation, principal component analysis, and cluster analysis. The IPR 85, IPR Catuara TM, T 091015, and T 091069 genotypes stood out from the others, which indicate their possibly superior grain quality with higher sedimentation volume, higher sedimentation index, and higher mass of 1000 seeds; these genotypes possessed the subunits 1 (Glu-A1, 5 + 10 (Glu-D1, c (Glu-A3, and b (Glu-B3, with exception of T 091069 genotype that possessed the g allele instead of b in the Glu-B3.

  19. Single-dose monomeric HA subunit vaccine generates full protection from influenza challenge

    CSIR Research Space (South Africa)

    Mallajosyula, JK

    2014-03-01

    Full Text Available Recombinant subunit vaccines are an efficient strategy to meet the demands of a possible influenza pandemic, because of rapid and scalable production. However, vaccines made from recombinant hemagglutinin (HA) subunit protein are often of low...

  20. Subunit composition and chromophore content of R-phycoerythrin from Porphyra haitanensis

    Science.gov (United States)

    Gao, Hong-Feng; Ji, Ming-Hou; Cao, Wen-Da

    1996-03-01

    R-phycoerythrin from Porphyra haitanensis exists in two aggregation states with different molecular weights. A more highly aggregated form, RPE I, was chromatographed on Bio-Rex 70 column with urea solution (pH 3.0) as eluent, and the molecular weights of the 3 subunits (α, β, γ) obtained were determined on SDS-PAGE at 18000, 19200 and 30000, respectively. α subunit carried two phycoerythrobilin (PEB); β subunit, three PEB and one phycourobilin (PUB); γ subunit, one PEB and three PUB chromophores. The molar ratio of α, β, and γ subunits of RPE I was 6: 6: 1, and their subunit composition was confired to be (αβ)6γ on account of the molecular weight of RPE I, 232000. A lower aggregated form, RPE II, contained α and β subunits similar to those of RPE I, but its subunit composition was the (αβ) monomer of RPE.

  1. Differential Contribution of Subunit Interfaces to α9α10 Nicotinic Acetylcholine Receptor Function

    Science.gov (United States)

    Boffi, Juan Carlos; Marcovich, Irina; Gill-Thind, JasKiran K.; Corradi, Jeremías; Collins, Toby; Lipovsek, María Marcela; Moglie, Marcelo; Plazas, Paola V.; Craig, Patricio O.; Millar, Neil S.; Bouzat, Cecilia

    2017-01-01

    Nicotinic acetylcholine receptors can be assembled from either homomeric or heteromeric pentameric subunit combinations. At the interface of the extracellular domains of adjacent subunits lies the acetylcholine binding site, composed of a principal component provided by one subunit and a complementary component of the adjacent subunit. Compared with neuronal nicotinic acetylcholine cholinergic receptors (nAChRs) assembled from α and β subunits, the α9α10 receptor is an atypical member of the family. It is a heteromeric receptor composed only of α subunits. Whereas mammalian α9 subunits can form functional homomeric α9 receptors, α10 subunits do not generate functional channels when expressed heterologously. Hence, it has been proposed that α10 might serve as a structural subunit, much like a β subunit of heteromeric nAChRs, providing only complementary components to the agonist binding site. Here, we have made use of site-directed mutagenesis to examine the contribution of subunit interface domains to α9α10 receptors by a combination of electrophysiological and radioligand binding studies. Characterization of receptors containing Y190T mutations revealed unexpectedly that both α9 and α10 subunits equally contribute to the principal components of the α9α10 nAChR. In addition, we have shown that the introduction of a W55T mutation impairs receptor binding and function in the rat α9 subunit but not in the α10 subunit, indicating that the contribution of α9 and α10 subunits to complementary components of the ligand-binding site is nonequivalent. We conclude that this asymmetry, which is supported by molecular docking studies, results from adaptive amino acid changes acquired only during the evolution of mammalian α10 subunits. PMID:28069778

  2. Advantages of a synthetic peptide immunogen over a protein immunogen in the development of an anti-pilus vaccine for Pseudomonas aeruginosa.

    Science.gov (United States)

    Kao, Daniel J; Hodges, Robert S

    2009-07-01

    The type IV pilus is an important adhesin in the establishment of infection by Pseudomonas aeruginosa. We have previously reported on a synthetic peptide vaccine targeting the receptor-binding domain of the main structural subunit of the pilus, PilA. The receptor-binding domain is a 14-residue disulfide loop at the C-terminal end of the pilin protein. The objective of this study was to compare the immunogenicity of a peptide-conjugate to a protein subunit immunogen to determine which was superior for use in an anti-pilus vaccine. BALB/c mice were immunized with the native PAK strain pilin protein and a synthetic peptide of the receptor-binding domain conjugated to keyhole limpet haemocyanin. A novel pilin protein with a scrambled receptor-binding domain was used to characterize receptor-binding domain-specific antibodies. The titres against the native pilin of the animals immunized with the synthetic peptide-conjugate were higher than the titres of animals immunized with the pilin protein. In addition, the affinities of anti-peptide sera for the intact pilin receptor-binding domain were significantly higher than affinities of anti-pilin protein sera. These results have significant implications for vaccine design and show that there are significant advantages in using a synthetic peptide-conjugate over a subunit pilin protein for an anti-pilus vaccine.

  3. Differential expression of G protein alpha and ß subunit genes during development of Phytophthora infestans

    NARCIS (Netherlands)

    Laxalt, A.M.; Latijnhouwers, M.; Hulten, van M.; Govers, F.

    2002-01-01

    A G protein subunit gene (pigpa1) and a G protein subunit gene (pigpb1) were isolated from the oomycete Phytophthora infestans, the causal agent of potato late blight. Heterotrimeric G proteins are evolutionary conserved GTP-binding proteins that are composed of ,, and subunits and participate in

  4. Testing experimental subunit furunculosis vaccines for rainbow trout

    DEFF Research Database (Denmark)

    Marana, Moonika H.; Chettri, Jiwan Kumar; Skov, Jakob

    2016-01-01

    Aeromonas salmonicida subsp. salmonicida (AS) is the etiological agent of typical furunculosis in salmonid fish. The disease causes bacterial septicemia and is a major fish health problem in salmonid aquaculture worldwide, inducing high morbidity and mortality. In this study we vaccinated rainbow...... trout with subunit vaccines containing protein antigens that were selected based on an in silico antigen discovery approach. Thus, the proteome of AS strain A449 was analyzed by an antigen discovery platform and its proteins consequently ranked by their predicted ability to evoke protective immune...... response against AS. Fourteen proteins were prepared in 3 different experimental subunit vaccine combinations and used to vaccinate rainbow trout by intraperitoneal (i.p.) injection. We tested the proteins for their ability to elicit antibody production and protection. Thus, fish were exposed to virulent...

  5. Architecture of the large subunit of the mammalian mitochondrial ribosome.

    Science.gov (United States)

    Greber, Basil J; Boehringer, Daniel; Leitner, Alexander; Bieri, Philipp; Voigts-Hoffmann, Felix; Erzberger, Jan P; Leibundgut, Marc; Aebersold, Ruedi; Ban, Nenad

    2014-01-23

    Mitochondrial ribosomes synthesize a number of highly hydrophobic proteins encoded on the genome of mitochondria, the organelles in eukaryotic cells that are responsible for energy conversion by oxidative phosphorylation. The ribosomes in mammalian mitochondria have undergone massive structural changes throughout their evolution, including ribosomal RNA shortening and acquisition of mitochondria-specific ribosomal proteins. Here we present the three-dimensional structure of the 39S large subunit of the porcine mitochondrial ribosome determined by cryo-electron microscopy at 4.9 Å resolution. The structure, combined with data from chemical crosslinking and mass spectrometry experiments, reveals the unique features of the 39S subunit at near-atomic resolution and provides detailed insight into the architecture of the polypeptide exit site. This region of the mitochondrial ribosome has been considerably remodelled compared to its bacterial counterpart, providing a specialized platform for the synthesis and membrane insertion of the highly hydrophobic protein components of the respiratory chain.

  6. Protein kinase A regulatory subunit distribution in medulloblastoma

    International Nuclear Information System (INIS)

    Mucignat-Caretta, Carla; Denaro, Luca; Redaelli, Marco; D'Avella, Domenico; Caretta, Antonio

    2010-01-01

    Previous studies showed a differential distribution of the four regulatory subunits of cAMP-dependent protein kinases inside the brain, that changed in rodent gliomas: therefore, the distribution of these proteins inside the brain can give information on the functional state of the cells. Our goal was to examine human brain tumors to provide evidence for a differential distribution of protein kinase A in different tumors. The distribution of detergent insoluble regulatory (R1 and R2) and catalytic subunits of cAMP dependent kinases was examined in pediatric brain tumors by immunohistochemistry and fluorescent cAMP analogues binding. R2 is organized in large single dots in medulloblastomas, while it has a different appearance in other tumors. Fluorescent cAMP labelling was observed only in medulloblastoma. A different distribution of cAMP dependent protein kinases has been observed in medulloblastoma

  7. ASIC subunit ratio and differential surface trafficking in the brain.

    Science.gov (United States)

    Wu, Junjun; Xu, Yuanyuan; Jiang, Yu-Qing; Xu, Jiangping; Hu, Youjia; Zha, Xiang-ming

    2016-01-08

    Acid-sensing ion channels (ASICs) are key mediators of acidosis-induced responses in neurons. However, little is known about the relative abundance of different ASIC subunits in the brain. Such data are fundamental for interpreting the relative contribution of ASIC1a homomers and 1a/2 heteromers to acid signaling, and essential for designing therapeutic interventions to target these channels. We used a simple biochemical approach and semi-quantitatively determined the molar ratio of ASIC1a and 2 subunits in mouse brain. Further, we investigated differential surface trafficking of ASIC1a, ASIC2a, and ASIC2b. ASIC1a subunits outnumber the sum of ASIC2a and ASIC2b. There is a region-specific variation in ASIC2a and 2b expression, with cerebellum and striatum expressing predominantly 2b and 2a, respectively. Further, we performed surface biotinylation and found that surface ASIC1a and ASIC2a ratio correlates with their total expression. In contrast, ASIC2b exhibits little surface presence in the brain. This result is consistent with increased co-localization of ASIC2b with an ER marker in 3T3 cells. Our data are the first semi-quantitative determination of relative subunit ratio of various ASICs in the brain. The differential surface trafficking of ASICs suggests that the main functional ASICs in the brain are ASIC1a homomers and 1a/2a heteromers. This finding provides important insights into the relative contribution of various ASIC complexes to acid signaling in neurons.

  8. Energetics of subunit assembly and ligand binding in human hemoglobin.

    OpenAIRE

    Ackers, G K

    1980-01-01

    An extensive and self-consistent set of thermodynamic properties has recently been established for the coupled processes of subunit assembly and ligand binding (oxygen and protons) in human hemoglobin. The resulting thermodynamic values permit a consideration of the possible sources of energetic terms accounting for stability of the tetrameric quaternary structures at different stages of ligation, and of the possible sources of cooperative energy. The analysis indicates that: (a) The change i...

  9. A fully automatic tool to perform accurate flood mapping by merging remote sensing imagery and ancillary data

    Science.gov (United States)

    D'Addabbo, Annarita; Refice, Alberto; Lovergine, Francesco; Pasquariello, Guido

    2016-04-01

    Flooding is one of the most frequent and expansive natural hazard. High-resolution flood mapping is an essential step in the monitoring and prevention of inundation hazard, both to gain insight into the processes involved in the generation of flooding events, and from the practical point of view of the precise assessment of inundated areas. Remote sensing data are recognized to be useful in this respect, thanks to the high resolution and regular revisit schedules of state-of-the-art satellites, moreover offering a synoptic overview of the extent of flooding. In particular, Synthetic Aperture Radar (SAR) data present several favorable characteristics for flood mapping, such as their relative insensitivity to the meteorological conditions during acquisitions, as well as the possibility of acquiring independently of solar illumination, thanks to the active nature of the radar sensors [1]. However, flood scenarios are typical examples of complex situations in which different factors have to be considered to provide accurate and robust interpretation of the situation on the ground: the presence of many land cover types, each one with a particular signature in presence of flood, requires modelling the behavior of different objects in the scene in order to associate them to flood or no flood conditions [2]. Generally, the fusion of multi-temporal, multi-sensor, multi-resolution and/or multi-platform Earth observation image data, together with other ancillary information, seems to have a key role in the pursuit of a consistent interpretation of complex scenes. In the case of flooding, distance from the river, terrain elevation, hydrologic information or some combination thereof can add useful information to remote sensing data. Suitable methods, able to manage and merge different kind of data, are so particularly needed. In this work, a fully automatic tool, based on Bayesian Networks (BNs) [3] and able to perform data fusion, is presented. It supplies flood maps

  10. Radioimmunoassay of TSH subunits in thyroid diseases and endocrine opthalmopahty

    International Nuclear Information System (INIS)

    Eder, W.

    1982-01-01

    Highly sensitive radioimmunoassays of hTSH sub-units were developed. The hormone preparations were labelled with 125-iodine according to a modified chloramine -T method, and purified by chromatography using biogel P6 and P60. Rabbit antisera were used as antibodies. Separation of the antibody-bound and of the free antigens was carried out via the double antibody method. The antiserum required for this purpose was obtained from a goat. The sensitivity of the assay was influenced by changing the protein content of the buffer, the incubation volume, the tracer amounts, the incubation time and the incubation temperature. For hTSH-α, the lowest detectable limit was found to be 50 pg/ml, for hTSH-#betta# 20 pg/ml. Thus, the sub-units could be determined for 98% of the patients under review. The #betta#-TSH radioimmunoassay is largely specific, TSH cross-reacts to a degree of 5%. The computerized evoluation was carried out by means of Spline approximation using the Siemens 4004 computer. Precision and accurateness are in compliance with generally accpted criteria. The serum levels of α and #betta# sub-units showed no discordancy with regard to TSH. In all groups of patients examined, the levels of the hormone-specific #betta#-chain were found to be exclusively dependent upon the actual thyroid activity. (orig.) [de

  11. The influence of different cyclometalated ligand substituents and ancillary ligand on the phosphorescent properties of iridium(III) complexes

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Qing; Li, Yuanyuan; Wang, Xin; Wang, Li, E-mail: chemwangl@henu.edu.cn; Zhang, Jinglai, E-mail: zhangjinglai@henu.edu.cn

    2016-07-01

    Four iridium(III) complexes, (dfpmpy){sub 2}Ir(pic), (1), (dfpmpy){sub 2}Ir(EO{sub 2}-pic) (2), (dfpmpy){sub 2}Ir(pic-N-O) (3), and (dfpmpy){sub 2}Ir(EO{sub 2}-pic-N-O) (4) (dfpmpy = 2-(2,4-difluorophenyl)-4-methylpyridine, pic = picolinic acid, EO{sub 2}-pic = 4-(2-ethoxyethoxy) picolinic acid, pic-N-O = picolinic acid N-oxide, and EO{sub 2}-pic-N-O = 4-(2-ethoxyethoxy) picolinic acid N-oxide) are investigated by means of the density functional theory/time-dependent density functional theory (DFT/TD-DFT) to explore the influence of the ancillary ligand on the electronic structures, phosphorescent properties, and organic light-emitting diode (OLED) performance. Employing pic-N-O and EO{sub 2}-pic-N-O as the ancillary ligands would decrease the vertical energy and result in the red-shifted wavelength. Then, other four iridium(III) complexes (2a-2d) (See Scheme 1) are designed by introduction of the phenyl and −CHO substituents on the pyridine ring and phenyl ring of complex 2, respectively. As compared with complex 2, theoretical results show that newly designed complexes 2a-2c might be potential candidates for blue-emitting phosphors with better/comparable quantum yield and Δλ. Moreover, the performance of complexes 2a and 2c, i.e., introducing phenyl on the para-position of pyridine ring and phenyl ring in dfpmpy ligand, are better than that of 2b. - Highlights: • The structure-property relationship of Ir(III) complexes are investigated. • The effect of different substituents/positions on properties is explored. • Do the emissions follow the Kasha or non-Kasha scenario? • Newly possible blue-emitting Ir(III) complexes are theoretically designed.

  12. Trends in OSHA Compliance Monitoring Data 1979-2011: Statistical Modeling of Ancillary Information across 77 Chemicals.

    Science.gov (United States)

    Sarazin, Philippe; Burstyn, Igor; Kincl, Laurel; Lavoué, Jérôme

    2016-05-01

    The Integrated Management Information System (IMIS) is the largest multi-industry source of exposure measurements available in North America. However, many have suspected that the criteria through which worksites are selected for inspection are related to exposure levels. We investigated associations between exposure levels and ancillary variables in IMIS in order to understand the predictors of high exposure within an enforcement context. We analyzed the association between nine variables (reason for inspection, establishment size, total amount of penalty, Occupational Safety and Health Administration (OSHA) plan, OSHA region, union status, inspection scope, year, and industry) and exposure levels in IMIS using multimodel inference for 77 agents. For each agent, we used two different types of models: (i) logistic models were used for the odds ratio (OR) of exposure being above the threshold limit value (TLV) and (ii) linear models were used for exposure concentrations restricted to detected results to estimate percent increase in exposure level, i.e. relative index of exposure (RIE). Meta-analytic methods were used to combine results for each variable across agents. A total of 511,047 exposure measurements were modeled for logistic models and 299,791 for linear models. Higher exposures were measured during follow-up inspections than planned inspections [meta-OR = 1.61, 95% confidence interval (CI): 1.44-1.81; meta-RIE = 1.06, 95% CI: 1.03-1.09]. Lower exposures were observed for measurements collected under state OSHA plans compared to measurements collected under federal OSHA (meta-OR = 0.82, 95% CI: 0.73-0.92; meta-RIE = 0.86, 95% CI: 0.81-0.91). A 'high' total historical amount of penalty relative to none was associated with higher exposures (meta-OR = 1.54, 95% CI: 1.40-1.71; meta-RIE = 1.18, 95% CI: 1.13-1.23). The relationships observed between exposure levels and ancillary variables across a vast majority of agents suggest that certain elements of OSHA

  13. On the multiple roles of the voltage gated sodium channel β1 subunit in genetic diseases

    Directory of Open Access Journals (Sweden)

    Debora eBaroni

    2015-05-01

    Full Text Available Voltage-gated sodium channels are intrinsic plasma membrane proteins that initiate the action potential in electrically excitable cells. They are composed of a pore-forming α-subunit and associated β-subunits. The β1-subunit was the first accessory subunit to be cloned. It can be important for controlling cell excitability and modulating multiple aspects of sodium channel physiology. Mutations of β1 are implicated in a wide variety of inherited pathologies, including epilepsy and cardiac conduction diseases. This review summarizes β1-subunit related channelopathies pointing out the current knowledge concerning their genetic background and their underlying molecular mechanisms.

  14. Subunits of the Schizosaccharomyces pombe RNA polymerase II: enzyme purification and structure of the subunit 3 gene.

    OpenAIRE

    Azuma, Y; Yamagishi, M; Ishihama, A

    1993-01-01

    To improve our understanding of the structure and function of eukaryotic RNA polymerase II, we purified the enzyme from the fission yeast Schizosaccharomyces pombe. The highly purified RNA polymerase II contained more than eleven polypeptides. The sizes of the largest the second-, and the third-largest polypeptides as measured by SDS-polyacrylamide gel electrophoresis were about 210, 150, and 40 kilodaltons (kDa), respectively, and are similar to those of RPB1, 2, and 3 subunits of Saccharomy...

  15. Subunit topography of RNA polymerase from Escherichia coli. A cross-linking study with bifunctional reagents.

    Science.gov (United States)

    Hillel, Z; Wu, C W

    1977-07-26

    The quaternary structures of Escherichia coli DNA-dependent RNA polymerase holenzyme (alpha 2 beta beta' sigma) and core enzyme (alpha 2 beta beta') have been investigated by chemical cross-linking with a cleavable bifunctional reagent, methyl 4-mercaptobutyrimidate, and noncleavable reagents, dimethyl suberimidate and N,N'-(1,4-phenylene)bismaleimide. A model of the subunit organization deduced from cross-linked subunit neighbors identified by dodecyl sulfate-polyacrylamide gel electrophoresis indicates that the large beta and beta' subunits constitute the backbone of both core and holoenzyme, while sigma and two alpha subunits interact with this structure along the contact domain of beta and beta' subunits. In holoenzyme, sigma subunit is in the vicinity of at least one alpha subunit. The two alpha subunits are close to each other in holoenzyme, core enzyme, and the isolated alpha 2 beta complex. Cross-linking of the "premature" core and holoenzyme intermediates in the in vitro reconstitution of active enzyme from isolated subunits suggests that these species are composed of subunit complexes of molecular weight lower than that of native core and holoenzyme, respectively. The structural information obtained for RNA polymerase and its subcomplexes has important implications for the enzyme-promoter recognition as well as the mechanism of subunit assembly of the enzyme.

  16. Prevalence and Correlates of Perceived Ethnic Discrimination in the Hispanic Community Health Study/Study of Latinos Sociocultural Ancillary Study.

    Science.gov (United States)

    Arellano-Morales, Leticia; Roesch, Scott C; Gallo, Linda C; Emory, Kristen T; Molina, Kristine M; Gonzalez, Patricia; Penedo, Frank J; Navas-Nacher, Elena L; Teng, Yanping; Deng, Yu; Isasi, Carmen R; Schneiderman, Neil; Brondolo, Elizabeth

    2015-08-01

    Empirical studies examining perceived ethnic discrimination in Latinos of diverse background groups are limited. This study examined prevalence and correlates of discrimination in a diverse sample of U.S. Latinos (N=5,291) from the multi-site Hispanic Community Health Study/Study of Latinos (HCHS/SOL) and HCHS/SOL Sociocultural Ancillary Study. The sample permitted an examination of differences across seven groups (Central American, Cuban, Dominican, Mexican, Puerto Rican, South American, and Other/Multiple Background). Most participants (79.5%) reported lifetime discrimination exposure and prevalence rates ranged from 64.9% to 98% across groups. Structural Equation Models (SEM) indicated that after adjusting for sociodemographic covariates most group differences in reports of discrimination were eliminated. However, Cubans reported the lowest levels of discrimination, overall among all groups. Furthermore, regional effects were more important than group effects. Participants from Chicago reported the highest levels of discrimination in comparison to other regions. Group differences among Latinos appear to be primarily a function of sociodemographic differences in education, income, and acculturation. In addition, differences in exposure to discrimination may be tied to variables associated with both immigration patterns and integration to U.S. culture. Results highlight the importance of considering historical context and the intersection of discrimination and immigration when evaluating the mental health of Latinos.

  17. Assistance in chemistry and chemical processes related to primary, secondary and ancillary systems of nuclear power plants

    International Nuclear Information System (INIS)

    Chocron, Mauricio A.; Becquart, Elena T.; Iglesias, Alberto M.; La Gamma, Ana M.; Villegas, Marina

    2003-01-01

    Argentina is currently running two nuclear power plants: Atucha I (CNA I) and Embalse (CNE) operated by Nucleoelectrica Argentina (NASA) whereas the National Atomic Energy Commission (CNEA), among other activities, is responsible for research and development in the nuclear field, operates research reactors and carries out projects related to them. In particular, the Reactor Chemistry Section personnel (currently part of the Chemistry Dept.) has been working on the field of reactor water chemistry for more than 25 years, on research and support to the NPPs chemistry department. Though the most relevant tasks have been connected to primary and secondary circuits chemistry, ancillary systems show along the time unexpected problems or feasible improvements originated in the undergoing operating time as well as in phenomena not foreseen by the constructors. In the present paper are presented the tasks performed in relation to the following systems of Embalse NPP: 1) Heavy water upgrade column preliminary water treatment; 2) Liquid waste system preliminary water treatment; and 3) Primary heat transport system coolant crud composition. (author)

  18. Psychosocial Factors in the Relationship between Socioeconomic Status and Cardiometabolic Risk: the HCHS/SOL Sociocultural Ancillary Study.

    Science.gov (United States)

    McCurley, Jessica L; Penedo, Frank; Roesch, Scott C; Isasi, Carmen R; Carnethon, Mercedes; Sotres-Alvarez, Daniela; Schneiderman, Neil; Gonzalez, Patricia; Chirinos, Diana A; Camacho, Alvaro; Teng, Yanping; Gallo, Linda C

    2017-08-01

    U.S. Hispanics/Latinos display a high prevalence of metabolic syndrome (MetSyn), a group of co-occurring cardiometabolic risk factors (abdominal obesity, impaired fasting glucose, dyslipidemia, elevated blood pressure) associated with higher cardiovascular disease and mortality risk. Low socioeconomic status (SES) is associated with higher risk for MetSyn in Hispanics/Latinos, and psychosocial factors may play a role in this relationship. This cross-sectional study examined psychosocial factors in the association of SES and MetSyn components in 4,996 Hispanic/Latino adults from the Hispanic Community Health Study/Study of Latinos (HCHS/SOL) Sociocultural Ancillary Study. MetSyn components were measured at the baseline examination. Participants completed interviews to determine psychosocial risks (e.g., depression) and resources (e.g., social support) within 9 months of baseline (psychosocial indicators, and three factors for MetSyn [blood pressure, lipids, metabolic factors]. SEMs showed that lower SES was related to MetSyn factors indirectly through higher psychosocial risk/lower resources (Y-Bχ 2 (df = 420) = 4412.90, p risk (glucose/waist circumference) via psychosocial risk/resource variables (Mackinnon's 95% asymmetric CI = -0.13 to -0.02). SES is related to metabolic variables indirectly through psychosocial factors in U.S. Hispanics/Latinos of diverse ancestries.

  19. DAFNE: A Matlab toolbox for Bayesian multi-source remote sensing and ancillary data fusion, with application to flood mapping

    Science.gov (United States)

    D'Addabbo, Annarita; Refice, Alberto; Lovergine, Francesco P.; Pasquariello, Guido

    2018-03-01

    High-resolution, remotely sensed images of the Earth surface have been proven to be of help in producing detailed flood maps, thanks to their synoptic overview of the flooded area and frequent revisits. However, flood scenarios can be complex situations, requiring the integration of different data in order to provide accurate and robust flood information. Several processing approaches have been recently proposed to efficiently combine and integrate heterogeneous information sources. In this paper, we introduce DAFNE, a Matlab®-based, open source toolbox, conceived to produce flood maps from remotely sensed and other ancillary information, through a data fusion approach. DAFNE is based on Bayesian Networks, and is composed of several independent modules, each one performing a different task. Multi-temporal and multi-sensor data can be easily handled, with the possibility of following the evolution of an event through multi-temporal output flood maps. Each DAFNE module can be easily modified or upgraded to meet different user needs. The DAFNE suite is presented together with an example of its application.

  20. 30+ New & Known SB2s in the SDSS-III/APOGEE M Dwarf Ancillary Science Project Sample

    Science.gov (United States)

    Skinner, Jacob; Covey, Kevin; Bender, Chad; De Lee, Nathan Michael; Chojnowski, Drew; Troup, Nicholas; Badenes, Carles; Mahadevan, Suvrath; Terrien, Ryan

    2018-01-01

    Close stellar binaries can drive dynamical interactions that affect the structure and evolution of planetary systems. Binary surveys indicate that the multiplicity fraction and typical orbital separation decrease with primary mass, but correlations with higher order architectural parameters such as the system's mass ratio are less well constrained. We seek to identify and characterize double-lined spectroscopic binaries (SB2s) among the 1350 M dwarf ancillary science targets with APOGEE spectra in the SDSS-III Data Release 13. We quantitatively measure the degree of asymmetry in the APOGEE pipeline cross-correlation functions (CCFs), and use those metrics to identify a sample of 44 high-likelihood candidate SB2s. Extracting radial velocities (RVs) for both binary components from the CCF, we then measure mass ratios for 31 SB2s; we also use Bayesian techniques to fit orbits for 4 systems with 8 or more distinct APOGEE observations. The (incomplete) mass ratio distribution of this sample rises quickly towards unity. Two-sided Kolmogorov-Smirnov (K-S) tests find probabilities of 13.8% and 14.2% that the M dwarf mass ratio distribution is consistent with those measured by Pourbaix et al. (2004) and Fernandez et al. (2017), respectively. The samples analyzed by Pourbaix et al. and Fernandez et al. are dominated by higher-mass solar type stars; this suggests that the mass ratio distribution of close binaries is not strongly dependent on primary mass.

  1. Engineering evaluation cost analysis for the 100-B/C area ancillary facilities at the 108-F Building

    International Nuclear Information System (INIS)

    1996-10-01

    In 1995, the US Department of Energy (DOE), Richland Operations Office (RL) conducted a removal site evaluation of selected facilities in the 100 Area of the Hanford Site in accordance with CERCLA and 40 Code of Federal Regulations (CFR) 300.410. The scope of the evaluation included the aboveground portions of the 108-F Biology Laboratory in the 100-F Area and all inactive ancillary buildings and structures in the 100-B/C Area, excluding the reactor building and the river outfall. Based on the evaluation, RL determined that hazardous substances in the 108-F Biology Laboratory and five of the 100-B/C Area facilities may present a potential threat to human health or the environment, and that a non-time critical removal action at these facilities is warranted. This determination was documented in an engineering evaluation/cost analysis (EE/CA) approval memorandum. The EE/CA approval memorandum is the basis on which to proceed with the performance of an EE/CA to determine the appropriate removal action. This report presents the results of the EE/CA for removal alternatives for final disposition of these six facilities. The EE/CA was conducted pursuant to the requirements of CERCLA and 40 CFR 300.415 and is intended to aid RL and the EPA in selecting a preferred removal action

  2. Chronic stress, inflammation, and glucose regulation in U.S. Hispanics from the HCHS/SOL Sociocultural Ancillary Study.

    Science.gov (United States)

    McCurley, Jessica L; Mills, Paul J; Roesch, Scott C; Carnethon, Mercedes; Giacinto, Rebeca E; Isasi, Carmen R; Teng, Yanping; Sotres-Alvarez, Daniela; Llabre, Maria M; Penedo, Frank J; Schneiderman, Neil; Gallo, Linda C

    2015-08-01

    Diabetes prevalence is rising rapidly, and diabetes disproportionately affects Hispanics and other underserved groups. Chronic stress may contribute to diabetes risk, but few studies have examined this relationship in U.S. Hispanics. We examined associations of chronic stress with fasting glucose, glucose tolerance, and glycosylated hemoglobin (HbA1c) in Hispanics without diabetes, and also assessed indirect effects of stress through inflammation (CRP). Participants were 3,923 men and women, aged 18-74, without diabetes, from the four U.S. field centers (Bronx, NY; Chicago, IL; Miami, FL; San Diego, CA) of the Hispanic Community Health Study/Study of Latinos (HCHS/SOL) Sociocultural Ancillary study. Participants completed a measure of chronic life stress and a physical exam with oral glucose tolerance test. In a multivariate regression analysis with adjustment for demographic and health covariates, higher chronic stress was related to higher fasting glucose (standardized regression coefficient: β = .09, p stress through inflammation. Findings suggest that higher chronic stress is associated with poorer glucose regulation in Hispanics, prior to the onset of a clinical diabetes diagnosis. © 2015 Society for Psychophysiological Research.

  3. Hostility and quality of life among Hispanics/Latinos in the HCHS/SOL Sociocultural Ancillary Study

    Science.gov (United States)

    Moncrieft, Ashley E.; Llabre, Maria M.; Gallo, Linda C.; Cai, Jianwen; Gonzalez, Franklyn; Gonzalez, Patricia; Ostrovsky, Natania W.; Schneiderman, Neil; Penedo, Frank J.

    2016-01-01

    Objective The purpose of this study was to determine if hostility is associated with physical and mental health-related quality of life (QoL) in U.S. Hispanics/Latinos after accounting for depression and anxiety. Methods Analyses included 5,313 adults (62% women, 18–75 years) who completed the ancillary sociocultural assessment of the Hispanic Community Health Study/Study of Latinos. Participants completed the Center for Epidemiological Studies Depression Scale, Spielberger Trait Anxiety Scale, Spielberger Trait Anger Scale, Cook-Medley Hostility cynicism subscale, and Short Form Health Survey. In a structural regression model, associations of hostility with mental and physical QoL were examined. Results In a model adjusting for age, sex, disease burden, income, education and years in the U.S., hostility was related to worse mental QoL, and was marginally associated with worse physical QoL. However, when adjusting for the influence of depression and anxiety, greater hostility was associated with better mental QoL, and was not associated with physical QoL. Conclusions Results indicate observed associations between hostility and QoL are confounded by symptoms of anxiety and depression, and suggest hostility is independently associated with better mental QoL in this population. Findings also highlight the importance of differentiating shared and unique associations of specific emotions with health outcomes. PMID:27456582

  4. Amino acid transporter B(0)AT1 (slc6a19) and ancillary protein: impact on function.

    Science.gov (United States)

    Margheritis, Eleonora; Imperiali, Francesca Guia; Cinquetti, Raffaella; Vollero, Alessandra; Terova, Genciana; Rimoldi, Simona; Girardello, Rossana; Bossi, Elena

    2016-08-01

    Amino acids play an important role in the metabolism of all organisms. Their epithelial re-absorption is due to specific transport proteins, such as B(0)AT1, a Na(+)-coupled neutral amino acid symporter belonging to the solute carrier 6 family. Here, a recently cloned fish orthologue, from the intestine of Salmo salar, was electrophysiologically characterized with the two-electrode voltage clamp technique, in Xenopus laevis oocytes heterologously expressing the transporter. Substrate specificity, apparent affinities and the ionic dependence of the transport mechanism were determined in the presence of specific collectrin. Results demonstrated that like the human, but differently from sea bass (Dicentrarchus labrax) orthologue, salmon B(0)AT1 needs to be associated with partner proteins to be correctly expressed at the oocyte plasma membrane. Cloning of sea bass collectrin and comparison of membrane expression and functionality of the B(0)AT1 orthologue transporters allowed a deeper investigation on the role of their interactions. The parameters acquired by electrophysiological and immunolocalization experiments in the mammalian and fish transporters contributed to highlight the dynamic of relations and impacts on transport function of the ancillary proteins. The comparative characterization of the physiological parameters of amino acid transporters with auxiliary proteins can help the comprehension of the regulatory mechanism of essential nutrient absorption.

  5. Proposal for the award of an industrial services contract for operation of the hostels, apartments and ancillary premises

    CERN Document Server

    2000-01-01

    This document concerns the award of an Industrial Services contract for operation of the hostels, apartments and ancillary premises run by the CERN Housing Service. Following a market survey carried out among 32 firms in ten Member States, a call for tenders (IT-2404/AS/Rev.) was sent on 5 April 2000 to one firm and three consortia in four Member States. By the closing date, CERN had received tenders from the firm and the three consortia. The Finance Committee is invited to agree to the negotiation of a contract with the consortium VIENNA INTERNATIONAL HOTEL MANAGEMENT (AT) - SEREG (CH), the lowest bidder, for an initial period of five years, to enter into force on 1 January 2001, for a total amount of 6 871 850 Swiss francs, not subject to revision until 31 December 2001. The contract will include an option for two one-year extensions beyond the initial five-year period. The consortium has indicated the following distribution by country of the contract value covered by this adjudication proposal: CH-60%, AT-...

  6. Effect of exercise training on clot strength in patients with peripheral artery disease and intermittent claudication: An ancillary study.

    Science.gov (United States)

    Mauer, Karin; Exaire, J Emilio; Stoner, Julie A; Saucedo, Jorge F; Montgomery, Polly S; Gardner, Andrew W

    2015-01-01

    Patients with peripheral artery disease have walking impairment, greater thrombotic risk, and are often treated with exercise training. We sought to determine the effect of a 3-month-long exercise program on clot strength among patients with peripheral artery disease and intermittent claudication. Twenty-three symptomatic peripheral artery disease patients were randomly assigned to a walking exercise program or to an attention control group who performed light resistance exercise. We investigated the effect of exercise training on clot strength and time to clot formation was assessed by thromboelastography. After 3 months of exercise, clot strength (maximal amplitude) and time to clot formation (R) did not change significantly from baseline, even after improvements in claudication onset time (p < 0.01) and peak walking time (p < 0.05). Furthermore, changes in clot formation parameters were not significantly different between groups. Among the 10 individuals demonstrating a reduction in clot strength (reduced maximal amplitude), one was a smoker (10%) compared to 9 of 13 non-responders (69%) whose maximal amplitude was unchanged or increased (p = 0.0097). In this ancillary study, a 12-week walking program improved ambulatory function in peripheral artery disease patients with claudication, but does not modify clot strength or time to clot formation. Larger studies are needed to confirm these hypothesis generating findings and to determine whether a different amount or type of exercise may induce a change in clotting in this patient population.

  7. Effect of exercise training on clot strength in patients with peripheral artery disease and intermittent claudication: An ancillary study

    Directory of Open Access Journals (Sweden)

    Karin Mauer

    2015-03-01

    Full Text Available Objectives: Patients with peripheral artery disease have walking impairment, greater thrombotic risk, and are often treated with exercise training. We sought to determine the effect of a 3-month-long exercise program on clot strength among patients with peripheral artery disease and intermittent claudication. Methods: Twenty-three symptomatic peripheral artery disease patients were randomly assigned to a walking exercise program or to an attention control group who performed light resistance exercise. We investigated the effect of exercise training on clot strength and time to clot formation was assessed by thromboelastography. Results: After 3 months of exercise, clot strength (maximal amplitude and time to clot formation (R did not change significantly from baseline, even after improvements in claudication onset time (p < 0.01 and peak walking time (p < 0.05. Furthermore, changes in clot formation parameters were not significantly different between groups. Among the 10 individuals demonstrating a reduction in clot strength (reduced maximal amplitude, one was a smoker (10% compared to 9 of 13 non-responders (69% whose maximal amplitude was unchanged or increased (p = 0.0097. Conclusion: In this ancillary study, a 12-week walking program improved ambulatory function in peripheral artery disease patients with claudication, but does not modify clot strength or time to clot formation. Larger studies are needed to confirm these hypothesis generating findings and to determine whether a different amount or type of exercise may induce a change in clotting in this patient population.

  8. Modelling Aṣṭādhyāyī: An Approach Based on the Methodology of Ancillary Disciplines (Vedāṅga)

    Science.gov (United States)

    Mishra, Anand

    This article proposes a general model based on the common methodological approach of the ancillary disciplines (Vedāṅga) associated with the Vedas taking examples from Śikṣā, Chandas, Vyākaraṇa and Prātiśā khya texts. It develops and elaborates this model further to represent the contents and processes of Aṣṭādhyāyī. Certain key features are added to my earlier modelling of Pāṇinian system of Sanskrit grammar. This includes broader coverage of the Pāṇinian meta-language, mechanism for automatic application of rules and positioning the grammatical system within the procedural complexes of ancillary disciplines.

  9. Synthesis and characterization of blue phosphorescent cyclometalated Ir(III) complexes containing 2-(imidazol-2-yl)pyridine as ancillary ligand

    Energy Technology Data Exchange (ETDEWEB)

    Xu Maoliang, E-mail: maoliangxu@yahoo.com.c [Xi' an Modern Chemistry Research Institute, Xi' an 710065 (China); Zhou Rui [Xi' an Modern Chemistry Research Institute, Xi' an 710065 (China); Che Guangbo [Department of Chemistry, Jilin Normal University, Siping 136000 (China); Wang Geyang; Wang Zijun; Xiao Qi [Xi' an Modern Chemistry Research Institute, Xi' an 710065 (China)

    2011-05-15

    Four new blue phosphorescent iridium complexes containing 2-(fluoro substituted phenyl)pyridine as the cyclometalated ligands and 2-(imidazol-2-yl)pyridine as an ancillary ligand have been synthesized and characterized. The complexes have the general structure (CcircumflexN){sub 2}Ir(pym), where CcircumflexN are cyclometalating ligands (e.g., 2-(2,4-difluorophenyl)pyridine (24f{sub 2}ppyH), 2-(3,4-difluorophenyl)pyridine (34f{sub 2}ppyH), 2-(3,5-difluorophenyl)pyridine (35f{sub 2}ppyH), and 2-(3,4,5-trifluorophenyl)pyridine (345f{sub 3}ppyH)), pym is 2-(imidazol-2-yl)pyridine (pymH). The absorption, emission, and cyclic voltammetry of the complexes were systematically investigated. The (46f{sub 2}ppy){sub 2}Ir(pym) has been characterized using X-ray crystallography and the electronic ground state calculated using B3LYP density functional theory. The highest occupied molecular orbital (HOMO) consists of a mixture of pym ligand (83.7%) and Ir orbitals (12.1%), while the lowest occupied molecular orbital (LUMO) is mostly on f{sub 2}ppy ligands (93.4%). By replacing the ancillary ligand pic with pym can finely tune emission of the iridium complexes, showing blue luminescence at a wavelength of 467-491 nm at room temperature in CH{sub 2}Cl{sub 2}. - Research highlights: {yields} Four new blue phosphorescent iridium complexes containing 2-(fluoro substituted phenyl)pyridine as the cyclometalated ligands and 2-(imidazol-2-yl)pyridine as an ancillary ligand have been synthesized and characterized. {yields} The absorption, emission, and cyclic voltammetry of the complexes were systematically investigated. {yields} By changing the ancillary ligand can finely tune the emission of the iridium complexes, which is a way to design and synthesize new phosphorescent iridium materials.

  10. Human mediator subunit MED15 promotes transcriptional activation.

    Science.gov (United States)

    Nakatsubo, Takuya; Nishitani, Saori; Kikuchi, Yuko; Iida, Satoshi; Yamada, Kana; Tanaka, Aki; Ohkuma, Yoshiaki

    2014-10-01

    In eukaryotes, the Mediator complex is an essential transcriptional cofactor of RNA polymerase II (Pol II). In humans, it contains up to 30 subunits and consists of four modules: head, middle, tail, and CDK/Cyclin. One of the subunits, MED15, is located in the tail module, and was initially identified as Gal11 in budding yeast, where it plays an essential role in the transcriptional regulation of galactose metabolism with the potent transcriptional activator Gal4. For this reason, we investigated the function of the human MED15 subunit (hMED15) in transcriptional activation. First, we measured the effect of hMED15 knockdown on cell growth in HeLa cells. The growth rate was greatly reduced. By immunostaining, we observed the colocalization of hMED15 with the general transcription factors TFIIE and TFIIH in the nucleus. We measured the effects of siRNA-mediated knockdown of hMED15 on transcriptional activation using two different transcriptional activators, VP16 and SREBP1a. Treatment with siRNAs reduced transcriptional activation, and this reduction could be rescued by overexpression of HA/Flag-tagged, wild-type hMED15. To investigate hMED15 localization, we treated human MCF-7 cells with the MDM2 inhibitor Nutlin-3, thus inducing p21 transcription. We found that hMED15 localized to both the p53 binding site and the p21 promoter region, along with TFIIE and TFIIH. These results indicate that hMED15 promotes transcriptional activation.

  11. Proposal for the award of a contract for the civil engineering work for the construction of the helium compressor buildings and the extension of the rectifier buildings and ancillary structures at LEP access points 4 and 8

    CERN Document Server

    1991-01-01

    Proposal for the award of a contract for the civil engineering work for the construction of the helium compressor buildings and the extension of the rectifier buildings and ancillary structures at LEP access points 4 and 8

  12. Crystal structure of the human Polϵ B-subunit in complex with the C-terminal domain of the catalytic subunit.

    Science.gov (United States)

    Baranovskiy, Andrey G; Gu, Jianyou; Babayeva, Nigar D; Kurinov, Igor; Pavlov, Youri I; Tahirov, Tahir H

    2017-09-22

    The eukaryotic B-family DNA polymerases include four members: Polα, Polδ, Polϵ, and Polζ, which share common architectural features, such as the exonuclease/polymerase and C-terminal domains (CTDs) of catalytic subunits bound to indispensable B-subunits, which serve as scaffolds that mediate interactions with other components of the replication machinery. Crystal structures for the B-subunits of Polα and Polδ/Polζ have been reported: the former within the primosome and separately with CTD and the latter with the N-terminal domain of the C-subunit. Here we present the crystal structure of the human Polϵ B-subunit (p59) in complex with CTD of the catalytic subunit (p261 C ). The structure revealed a well defined electron density for p261 C and the phosphodiesterase and oligonucleotide/oligosaccharide-binding domains of p59. However, electron density was missing for the p59 N-terminal domain and for the linker connecting it to the phosphodiesterase domain. Similar to Polα, p261 C of Polϵ contains a three-helix bundle in the middle and zinc-binding modules on each side. Intersubunit interactions involving 11 hydrogen bonds and numerous hydrophobic contacts account for stable complex formation with a buried surface area of 3094 Å 2 Comparative structural analysis of p59-p261 C with the corresponding Polα complex revealed significant differences between the B-subunits and CTDs, as well as their interaction interfaces. The B-subunit of Polδ/Polζ also substantially differs from B-subunits of either Polα or Polϵ. This work provides a structural basis to explain biochemical and genetic data on the importance of B-subunit integrity in replisome function in vivo . © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  13. Structure of protein kinase CK2: dimerization of the human beta-subunit

    DEFF Research Database (Denmark)

    Boldyreff, B; Mietens, U; Issinger, O G

    1996-01-01

    Protein kinase CK2 has been shown to be elevated in all so far investigated solid tumors and its catalytic subunit has been shown to serve as an oncogene product. CK2 is a heterotetrameric serine-threonine kinase composed of two catalytic (alpha and/or alpha') and two regulatory beta-subunits. Us......Protein kinase CK2 has been shown to be elevated in all so far investigated solid tumors and its catalytic subunit has been shown to serve as an oncogene product. CK2 is a heterotetrameric serine-threonine kinase composed of two catalytic (alpha and/or alpha') and two regulatory beta......-subunits. Using the two-hybrid system we could show that the alpha- or alpha'-subunits of CK2 can interact with the beta-subunits of CK2, but not with other alpha- or alpha'-subunits. By comparison, the beta-subunit of CK2 can interact with another beta-subunit. Important amino acids for successful dimerization...... of the beta-subunit were localized between amino acid residues 156 and 165. Furthermore, we identified residues between amino acid 170 and 180 which antagonize the dimerization....

  14. N-linked glycans are required on epithelial Na+ channel subunits for maturation and surface expression.

    Science.gov (United States)

    Kashlan, Ossama B; Kinlough, Carol L; Myerburg, Michael M; Shi, Shujie; Chen, Jingxin; Blobner, Brandon M; Buck, Teresa M; Brodsky, Jeffrey L; Hughey, Rebecca P; Kleyman, Thomas R

    2018-03-01

    Epithelial Na + channel (ENaC) subunits undergo N-linked glycosylation in the endoplasmic reticulum where they assemble into an αβγ complex. Six, 13, and 5 consensus sites (Asn-X-Ser/Thr) for N-glycosylation reside in the extracellular domains of the mouse α-, β-, and γ-subunits, respectively. Because the importance of ENaC N-linked glycans has not been fully addressed, we examined the effect of preventing N-glycosylation of specific subunits on channel function, expression, maturation, and folding. Heterologous expression in Xenopus oocytes or Fischer rat thyroid cells with αβγ-ENaC lacking N-linked glycans on a single subunit reduced ENaC activity as well as the inhibitory response to extracellular Na + . The lack of N-linked glycans on the β-subunit also precluded channel activation by trypsin. However, channel activation by shear stress was N-linked glycan independent, regardless of which subunit was modified. We also discovered that the lack of N-linked glycans on any one subunit reduced the total and surface levels of cognate subunits. The lack of N-linked glycans on the β-subunit had the largest effect on total levels, with the lack of N-linked glycans on the γ- and α-subunits having intermediate and modest effects, respectively. Finally, channels with wild-type β-subunits were more sensitive to limited trypsin proteolysis than channels lacking N-linked glycans on the β-subunit. Our results indicate that N-linked glycans on each subunit are required for proper folding, maturation, surface expression, and function of the channel.

  15. Stable incorporation versus dynamic exchange of β subunits in a native Ca2+ channel complex.

    Science.gov (United States)

    Campiglio, Marta; Di Biase, Valentina; Tuluc, Petronel; Flucher, Bernhard E

    2013-05-01

    Voltage-gated Ca(2+) channels are multi-subunit membrane proteins that transduce depolarization into cellular functions such as excitation-contraction coupling in muscle or neurotransmitter release in neurons. The auxiliary β subunits function in membrane targeting of the channel and modulation of its gating properties. However, whether β subunits can reversibly interact with, and thus differentially modulate, channels in the membrane is still unresolved. In the present study we applied fluorescence recovery after photobleaching (FRAP) of GFP-tagged α1 and β subunits expressed in dysgenic myotubes to study the relative dynamics of these Ca(2+) channel subunits for the first time in a native functional signaling complex. Identical fluorescence recovery rates of both subunits indicate stable interactions, distinct recovery rates indicate dynamic interactions. Whereas the skeletal muscle β1a isoform formed stable complexes with CaV1.1 and CaV1.2, the non-skeletal muscle β2a and β4b isoforms dynamically interacted with both α1 subunits. Neither replacing the I-II loop of CaV1.1 with that of CaV2.1, nor deletions in the proximal I-II loop, known to change the orientation of β relative to the α1 subunit, altered the specific dynamic properties of the β subunits. In contrast, a single residue substitution in the α interaction pocket of β1aM293A increased the FRAP rate threefold. Taken together, these findings indicate that in skeletal muscle triads the homologous β1a subunit forms a stable complex, whereas the heterologous β2a and β4b subunits form dynamic complexes with the Ca(2+) channel. The distinct binding properties are not determined by differences in the I-II loop sequences of the α1 subunits, but are intrinsic properties of the β subunit isoforms.

  16. Routine use of ancillary investigations in staging diffuse large B-cell lymphoma improves the International Prognostic Index (IPI).

    Science.gov (United States)

    Talaulikar, Dipti; Shadbolt, Bruce; Dahlstrom, Jane E; McDonald, Anne

    2009-11-22

    The International Prognostic Index (IPI) is used to determine prognosis in diffuse large B-cell lymphoma (DLBCL). One of the determinants of IPI is the stage of disease with bone marrow involvement being classified as stage IV. For the IPI, involvement on bone marrow is traditionally defined on the basis of histology with ancillary investigations used only in difficult cases to aid histological diagnosis. This study aimed to determine the effect of the routine use of flow cytometry, immunohistochemistry and molecular studies in bone marrow staging upon the IPI. Bone marrow trephines of 156 histologically proven DLBCL cases at initial diagnosis were assessed on routine histology, and immunohistochemistry using two T-cell markers (CD45RO and CD3), two B-cell markers (CD20 and CD79a) and kappa and lambda light chains. Raw flow cytometry data on all samples were reanalysed and reinterpreted blindly. DNA extracted from archived paraffin-embedded trephine biopsy samples was used for immunoglobulin heavy chain and light chain gene rearrangement analysis. Using immunophenotyping (flow cytometry and immunohistochemistry), 30 (19.2%) cases were upstaged to stage IV. A further 8 (5.1%) cases were upstaged using molecular studies. A change in IPI was noted in 18 cases (11.5%) on immunophenotyping alone, and 22 (14.1%) cases on immunophenotyping and molecular testing. Comparison of two revised IPI models, 1) using immunophenotyping alone, and 2) using immunophenotyping with molecular studies, was performed with baseline IPI using a Cox regression model. It showed that the revised IPI model using immunophenotyping provides the best differentiation between the IPI categories. Improved bone marrow staging using flow cytometry and immunohistochemistry improves the predictive value of the IPI in patients with DLBCL and should be performed routinely in all cases.

  17. Stochastic scheduling of aggregators of plug-in electric vehicles for participation in energy and ancillary service markets

    International Nuclear Information System (INIS)

    Alipour, Manijeh; Mohammadi-Ivatloo, Behnam; Moradi-Dalvand, Mohammad; Zare, Kazem

    2017-01-01

    Plug-in electric vehicles are expected to play a major role in the transportation system as the environmental problems and energy crisis are being more and more urgent recently. Implementing a large number of vehicles with proper control could bring an opportunity of large storage and flexibility for power systems. The plug-in electric vehicle aggregator is responsible for providing power and controlling the charging pattern of the plug-in electric vehicles under its contracted area. This paper deals with the problem of optimal scheduling problem of plug-in electric vehicle aggregators in electricity market considering the uncertainties of market prices, availability of vehicles and status of being called by the ISO in the reserve market. The impact of the market price and reserve market uncertainties on the electric vehicle scheduling problem is characterized through a stochastic programming framework. The objective of the aggregator is to maximize its profit by charging the plug-in electric vehicles on the low price time intervals as well as participating in ancillary service markets. The operational constraints of plug-in electric vehicles and constraints of vehicle to grid are modeled in the proposed framework. An illustrative example is provided to confirm the performance of the proposed model. - Highlights: • Optimal scheduling of vehicle aggregators in electricity market has been addressed. • The operational constraints of plug-in vehicle to grid are considered. • The uncertainties of calling status in reserve market and market prices are modeled. • Vehicles' driving patterns and availability uncertainty are modeled. • The effect of risk measure weight in the vehicle to grid model has been studied.

  18. Stress and sleep: Results from the Hispanic Community Health Study/Study of Latinos Sociocultural Ancillary Study

    Directory of Open Access Journals (Sweden)

    Carmela Alcántara

    2017-12-01

    Full Text Available Hispanics/Latinos face specific sociocultural stressors associated with their marginalized status in the United States. While stress is known to cause poor sleep, the differential effects of the specific stressors faced by Hispanics/Latinos have not been evaluated. Using cross-sectional data from the Hispanic Community Health Study/Study of Latinos Sociocultural Ancillary Study, we conducted weighted generalized linear models to evaluate the associations of acculturation stress, ethnic discrimination, and chronic moderate/severe stress with self-reported sleep outcomes (insomnia symptoms, daytime sleepiness, sleep duration in individual and aggregate models adjusted for site, socio-demographics, behavioral, and medical conditions. Participants included 5313 Hispanic/Latino adults; 43.5% ≥ age 45, 54.8% female, and 22.0% US-born. Chronic moderate/severe stress, ethnic discrimination, and acculturation stress were each positively associated with sleep. In the adjusted aggregate model, only chronic moderate/severe stress was associated with insomnia symptoms (exp(b = 1.07 for each additional stressor, 95% CI = 1.05, 1.09. Both acculturation stress (exp(b = 1.05 for each additional SD, 95% CI = 1.02, 1.10 and ethnic discrimination (exp(b = 1.05 for each additional SD, 95% CI = 1.01, 1.08 were associated with daytime sleepiness. Each SD increase in ethnic discrimination related to a 16% and 13% increased prevalence of short ( 9 h (RRR = 1.13, 95% CI = 1.00, 1.27, respectively. These associations were consistent across sex. Acculturation stress and ethnic discrimination are associated with poor sleep in Hispanics/Latinos. Future research should explore whether behavioral sleep interventions minimize the impact of sociocultural stressors on sleep.

  19. Land Cover Classification in an Ecuadorian Mountain Geosystem Using a Random Forest Classifier, Spectral Vegetation Indices, and Ancillary Geographic Data

    Directory of Open Access Journals (Sweden)

    Johanna E. Ayala-Izurieta

    2017-05-01

    Full Text Available We presented a methodology to accurately classify mountainous regions in the tropics. These landscapes are complex in terms of their geology, ecosystems, climate and land use. Obtaining accurate maps to assess land cover change is essential. The objectives of this study were to (1 map vegetation using the Random Forest Classifier (RFC, spectral vegetation index (SVI, and ancillar geographic data (2 identify important variables that help differentiate vegetation cover, and (3 assess the accuracy of the vegetation cover classification in hard-to-reach Ecuadorian mountain region. We used Landsat 7 ETM+ satellite images of the entire scene, a RFC algorithm, and stratified random sampling. The altitude and the two band enhanced vegetation index (EVI2 provide more information on vegetation cover than the traditional and often use normalized difference vegetation index (NDVI in other settings. We classified the vegetation cover of mountainous areas within the 1016 km2 area of study, at 30 m spatial resolution, using RFC that yielded a land cover map with an overall accuracy of 95%. The user´s accuracy and the half-width of the confidence interval for 95% of the basic map units, forest (FOR, páramo (PAR, crop (CRO and pasture (PAS were 95.85% ± 2.86%, 97.64% ± 1.24%, 91.53% ± 3.35% and 82.82% ± 7.74%, respectively. The overall disagreement was 4.47%, which results from adding 0.43% of quantity disagreement and 4.04% of allocation disagreement. The methodological framework presented in this paper and the combined use of SVIs, ancillary geographic data, and the RFC allowed the accurate mapping of hard-to-reach mountain landscapes as well as uncovering the underlying factors that help differentiate vegetation cover in the Ecuadorian mountain geosystem.

  20. Controllable Grid Interface for Testing Ancillary Service Controls and Fault Performance of Utility-Scale Wind Power Generation: Preprint

    Energy Technology Data Exchange (ETDEWEB)

    Gevorgian, Vahan; Koralewicz, Przemyslaw; Wallen, Robb; Muljadi, Eduard

    2017-02-01

    The rapid expansion of wind power has led many transmission system operators to demand modern wind power plants to comply with strict interconnection requirements. Such requirements involve various aspects of wind power plant operation, including fault ride-through and power quality performance as well as the provision of ancillary services to enhance grid reliability. During recent years, the National Renewable Energy Laboratory (NREL) of the U.S. Department of Energy has developed a new, groundbreaking testing apparatus and methodology to test and demonstrate many existing and future advanced controls for wind generation (and other renewable generation technologies) on the multimegawatt scale and medium-voltage levels. This paper describes the capabilities and control features of NREL's 7-MVA power electronic grid simulator (also called a controllable grid interface, or CGI) that enables testing many active and reactive power control features of modern wind turbine generators -- including inertial response, primary and secondary frequency responses, and voltage regulation -- under a controlled, medium-voltage grid environment. In particular, this paper focuses on the specifics of testing the balanced and unbalanced fault ride-through characteristics of wind turbine generators under simulated strong and weak medium-voltage grid conditions. In addition, this paper provides insights on the power hardware-in-the-loop feature implemented in the CGI to emulate (in real time) the conditions that might exist in various types of electric power systems under normal operations and/or contingency scenarios. Using actual test examples and simulation results, this paper describes the value of CGI as an ultimate modeling validation tool for all types of 'grid-friendly' controls by wind generation.

  1. Measurement and charactereistics of hCG β-subunit

    International Nuclear Information System (INIS)

    Isojima, Shinzo; Koyama, Koji; Naga, Osamu.

    1975-01-01

    The authors studied an hCG-radioimmunoassay method in which the anti-hCG β-subunit antibody- 125 I-hCG β-subunit system was used, and they determined that in this method the antibody reacted much more specifically against hCG than in the usual radioimmunoassay method in which the anti-hCG antibody- 125 I-hCG has been used. In addition, they explained that a specific method for hCG-radioimmunoassay can be established in which the reaction system does not depend on the amount of physiological LH secretion. The significance of the clinical application of this method was considered and it was proved that the method was necessary, especially for studies of the hCG secretion pattern in patients with villous tumors having a low level of LH. Furthermore, the phenomenon of the cellular effect, namely a temporary increase of the hCG value which was thought to be caused by the administration of anticancerous agents, was proved to be due to hCG hypersecretion, not merely to LH variation. (Ichikawa, K.)

  2. Thermostable cross-protective subunit vaccine against Brucella species.

    Science.gov (United States)

    Cherwonogrodzky, John W; Barabé, Nicole D; Grigat, Michelle L; Lee, William E; Poirier, Robert T; Jager, Scott J; Berger, Bradley J

    2014-12-01

    A subunit vaccine candidate was produced from Brucella suis 145 (biovar 4; expressing both the A antigen of Brucella abortus and the M antigen of Brucella melitensis). The preparation consisted mostly of polysaccharide (PS; >90% [wt/wt]; both cell-associated PS and exo-PS were combined) and a small amount of protein (1 to 3%) with no apparent nucleic acids. Vaccinated mice were protected (these had a statistically significant reduction in bacterial colonization compared to that of unvaccinated controls) when challenged with representative strains of three Brucella species most pathogenic for humans, i.e., B. abortus, B. melitensis, and B. suis. As little as 1 ng of the vaccine, without added adjuvant, protected mice against B. suis 145 infection (5 × 10(5) CFU), and a single injection of 1 μg of this subunit vaccine protected mice from B. suis 145 challenge for at least 14 months. A single immunization induced a serum IgG response to Brucella antigens that remained elevated for up to 9 weeks. The use of heat (i.e., boiling-water bath, autoclaving) in the vaccine preparation showed that it was thermostable. This method also ensured safety and security. The vaccine produced was immunogenic and highly protective against multiple strains of Brucella and represents a promising candidate for further evaluation. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  3. The TFIIH subunit Tfb3 regulates cullin neddylation

    Science.gov (United States)

    Rabut, Gwenaël; Le Dez, Gaëlle; Verma, Rati; Makhnevych, Taras; Knebel, Axel; Kurz, Thimo; Boone, Charles; Deshaies, Raymond J.; Peter, Matthias

    2011-01-01

    Summary Cullin proteins are scaffolds for the assembly of multi-subunit ubiquitin ligases, which ubiquitylate a large number of proteins involved in widely-varying cellular functions. Multiple mechanisms cooperate to regulate cullin activity, including neddylation of their C-terminal domain. Interestingly, we found that the yeast Cul4-type cullin Rtt101 is not only neddylated but also ubiquitylated, and both modifications promote Rtt101 function in vivo. Surprisingly, proper modification of Rtt101 neither correlated with catalytic activity of the RING-domain of Hrt1 nor did it require the Nedd8 ligase Dcn1. Instead, ubiquitylation of Rtt101 was dependent on the ubiquitin-conjugating enzyme Ubc4, while efficient neddylation involves the RING-domain protein Tfb3, a subunit of the transcription factor TFIIH. Tfb3 also controls Cul3 neddylation and activity in vivo, and physically interacts with Ubc4 and the Nedd8-conjugating enzyme Ubc12 as well as the Hrt1/Rtt101 complex. Together, these results suggest that the conserved RING-domain protein Tfb3 controls activation of a subset of cullins. PMID:21816351

  4. Binding of ATP by pertussis toxin and isolated toxin subunits

    International Nuclear Information System (INIS)

    Hausman, S.Z.; Manclark, C.R.; Burns, D.L.

    1990-01-01

    The binding of ATP to pertussis toxin and its components, the A subunit and B oligomer, was investigated. Whereas, radiolabeled ATP bound to the B oligomer and pertussis toxin, no binding to the A subunit was observed. The binding of [ 3 H]ATP to pertussis toxin and the B oligomer was inhibited by nucleotides. The relative effectiveness of the nucleotides was shown to be ATP > GTP > CTP > TTP for pertussis toxin and ATP > GTP > TTP > CTP for the B oligomer. Phosphate ions inhibited the binding of [ 3 H]ATP to pertussis toxin in a competitive manner; however, the presence of phosphate ions was essential for binding of ATP to the B oligomer. The toxin substrate, NAD, did not affect the binding of [ 3 H]ATP to pertussis toxin, although the glycoprotein fetuin significantly decreased binding. These results suggest that the binding site for ATP is located on the B oligomer and is distinct from the enzymatically active site but may be located near the eukaryotic receptor binding site

  5. Binding of ATP by pertussis toxin and isolated toxin subunits

    Energy Technology Data Exchange (ETDEWEB)

    Hausman, S.Z.; Manclark, C.R.; Burns, D.L. (Center for Biologics Evaluation and Research, Bethesda, MD (USA))

    1990-07-03

    The binding of ATP to pertussis toxin and its components, the A subunit and B oligomer, was investigated. Whereas, radiolabeled ATP bound to the B oligomer and pertussis toxin, no binding to the A subunit was observed. The binding of ({sup 3}H)ATP to pertussis toxin and the B oligomer was inhibited by nucleotides. The relative effectiveness of the nucleotides was shown to be ATP > GTP > CTP > TTP for pertussis toxin and ATP > GTP > TTP > CTP for the B oligomer. Phosphate ions inhibited the binding of ({sup 3}H)ATP to pertussis toxin in a competitive manner; however, the presence of phosphate ions was essential for binding of ATP to the B oligomer. The toxin substrate, NAD, did not affect the binding of ({sup 3}H)ATP to pertussis toxin, although the glycoprotein fetuin significantly decreased binding. These results suggest that the binding site for ATP is located on the B oligomer and is distinct from the enzymatically active site but may be located near the eukaryotic receptor binding site.

  6. Kinetic mechanism of luciferase subunit folding and assembly.

    Science.gov (United States)

    Clark, A C; Raso, S W; Sinclair, J F; Ziegler, M M; Chaffotte, A F; Baldwin, T O

    1997-02-18

    The kinetic mechanism in vitro of the folding and assembly of the heterodimeric flavin monooxygenase bacterial luciferase has been defined by a unique set of rate constants which describe both the productive refolding pathway and competing off-pathway reactions in 50 mM phosphate, pH 7.0 at 18 degrees C. The individual alpha and beta subunits fold independently to form heterodimerization-competent species, alpha i and beta i. The alpha i beta i species can interact to form an inactive heterodimeric intermediate, [alpha beta ]i, which isomerizes to form the active alpha beta structure; the structure of the enzyme has been determined to 1.5 A resolution [Fisher, A. J., Thompson, T. B., Thoden, J. B., Baldwin, T. O., & Rayment, I. (1996) J. Biol. Chem. 271, 21956-21968]. In the absence of alpha i, beta i can form a kinetically trapped homodimer, beta 2, with a second-order rate constant of about 180 M-1 s-1 [Sinclair, J. F., Ziegler, M. M., & Baldwin, T. O. (1994) Nat. Struct. Biol. 1, 320-326]; the structure of beta 2 has recently been reported [Thoden. J. B., Holden, H. M., Fisher, A. J., Sinclair. J. F., Wesenberg, G., Baldwin, T.O., & Rayment, I. (1997) Protein Sci. 6, 13-23]. The beta i species, or some other form that precedes beta i on the refolding pathway, can also undergo a first-order conversion into a form (designated beta x) that cannot associate with alpha i to form the native enzyme. The rate constant for this process, assigned here, accounts well for the previously observed dependence of final yield on concentration of refolding species [Ziegler, M.M., Goldberg, M.E., Chaffotte, A. F., & Baldwin, T. O. (1993) J. Biol. Chem. 268, 10760-10765]. In simulations of the refolding reaction, all processes associated with the refolding of the individual subunits were combined into single first-order rate constants for each subunit which were consistent with the rate constants determined from stopped-flow circular dichroism studies. The first-order rate constant

  7. Efficient expression of functional (α6β22β3 AChRs in Xenopus oocytes from free subunits using slightly modified α6 subunits.

    Directory of Open Access Journals (Sweden)

    Carson Kai-Kwong Ley

    Full Text Available Human (α6β2(α4β2β3 nicotinic acetylcholine receptors (AChRs are essential for addiction to nicotine and a target for drug development for smoking cessation. Expressing this complex AChR is difficult, but has been achieved using subunit concatamers. In order to determine what limits expression of α6* AChRs and to efficiently express α6* AChRs using free subunits, we investigated expression of the simpler (α6β22β3 AChR. The concatameric form of this AChR assembles well, but is transported to the cell surface inefficiently. Various chimeras of α6 with the closely related α3 subunit increased expression efficiency with free subunits and produced pharmacologically equivalent functional AChRs. A chimera in which the large cytoplasmic domain of α6 was replaced with that of α3 increased assembly with β2 subunits and transport of AChRs to the oocyte surface. Another chimera replacing the unique methionine 211 of α6 with leucine found at this position in transmembrane domain 1 of α3 and other α subunits increased assembly of mature subunits containing β3 subunits within oocytes. Combining both α3 sequences in an α6 chimera increased expression of functional (α6β22β3 AChRs to 12-fold more than with concatamers. This is pragmatically useful, and provides insights on features of α6 subunit structure that limit its expression in transfected cells.

  8. The modulator protein dissociates the catalytic subunit of hepatic protein phosphatase G from glycogen.

    OpenAIRE

    Bollen, M; Stalmans, W

    1988-01-01

    1. The phosphorylase phosphatase and glycogen-synthase phosphatase activities associated with the glycogen particles from rat liver were progressively inhibited by incubation with modulator protein. However, the phosphorylase phosphatase activity of the catalytic subunit was entirely recovered after destruction of the modulator and the regulatory subunit(s) by trypsin. 2. Inhibition of protein phosphatase G by modulator was associated with a translocation of the phosphorylase phosphatase acti...

  9. Compensatory expression of human -Acetylglucosaminyl-1-phosphotransferase subunits in mucolipidosis type III gamma

    OpenAIRE

    Pohl , Sandra; Tiede , Stephan; Castrichini , Monica; Cantz , Michael; Gieselmann , Volkmar; Braulke , Thomas

    2009-01-01

    Abstract The N-Acetylglucosaminyl-1-phosphotransferase plays a key role in the generation of mannose 6-phosphate (M6P) recognition markers essential for efficient transport of lysosomal hydrolases to lysosomes. The phosphotransferase is composed of six subunits (?2, ?2, ?2). The ?- and ?-subunits are catalytically active and encoded by a single gene, GNPTAB, whereas the ?-subunit encoded by GNPTG is proposed to recognize conformational structures common to lysosomal enzymes. Defects in GN...

  10. Cloning and sequencing of the casein kinase 2 alpha subunit from Zea mays

    DEFF Research Database (Denmark)

    Dobrowolska, G; Boldyreff, B; Issinger, O G

    1991-01-01

    The nucleotide sequence of the cDNA coding for the alpha subunit of casein kinase 2 of Zea mays has been determined. The cDNA clone contains an open reading frame of 996 nucleotides encoding a polypeptide comprising 332 amino acids. The primary amino acid sequence exhibits 75% identity to the alpha...... subunit and 71% identity to the alpha' subunit of human casein kinase 2....

  11. Beta-Subunit of Human Chorionic Gonadotropin in Malignant Lymphoma : An Immunohistochemical Study

    OpenAIRE

    Senba, Masachika; Watanabe, Masami

    1991-01-01

    We present a rare case of a 77-year-old Japanese man with malignant lymphoma associated with production of beta-subunit of human chorionic gonadotropin in the cytoplasms of lymphoma cells in the lymph nodes. By immunoperoxidase staining, numerous tumor cells were reacted with beta-subunit of human chorionic gonadotropin. To the best of our knowledge, production of beta-subunit of human chorionic gonadotropin in the cytoplasm of lymphoma cells has not been reported. This patient evidences that...

  12. Characterisation by nuclear magnetic resonance of the β catalytic subunit of the chloroplastic coupling factor

    International Nuclear Information System (INIS)

    Andre, Francois

    1986-09-01

    This academic work addressed the use of nuclear magnetic resonance (NMR) for the structural and dynamic study of the catalytic sub-unit of the extrinsic section of a membrane complex, the chloroplastic H+-ATPase. This work included the development of a protocol of preparation and quantitative purification of β subunits isolated from the CF1 for the elaboration of a concentrated sample for NMR, and then the study of the β subunit by using proton NMR

  13. The NH2-terminal php domain of the alpha subunit of the Escherichia coli replicase binds the epsilon proofreading subunit.

    Science.gov (United States)

    Wieczorek, Anna; McHenry, Charles S

    2006-05-05

    The alpha subunit of the replicase of all bacteria contains a php domain, initially identified by its similarity to histidinol phosphatase but of otherwise unknown function (Aravind, L., and Koonin, E. V. (1998) Nucleic Acids Res. 26, 3746-3752). Deletion of 60 residues from the NH2 terminus of the alpha php domain destroys epsilon binding. The minimal 255-residue php domain, estimated by sequence alignment with homolog YcdX, is insufficient for epsilon binding. However, a 320-residue segment including sequences that immediately precede the polymerase domain binds epsilon with the same affinity as the 1160-residue full-length alpha subunit. A subset of mutations of a conserved acidic residue (Asp43 in Escherichia coli alpha) present in the php domain of all bacterial replicases resulted in defects in epsilon binding. Using sequence alignments, we show that the prototypical gram+ Pol C, which contains the polymerase and proofreading activities within the same polypeptide chain, has an epsilon-like sequence inserted in a surface loop near the center of the homologous YcdX protein. These findings suggest that the php domain serves as a platform to enable coordination of proofreading and polymerase activities during chromosomal replication.

  14. Translation activity of chimeric ribosomes composed of Escherichia coli and Bacillus subtilis or Geobacillus stearothermophilus subunits

    Directory of Open Access Journals (Sweden)

    Sayaka Tsuji

    2017-07-01

    Full Text Available Ribosome composition, consisting of rRNA and ribosomal proteins, is highly conserved among a broad range of organisms. However, biochemical studies focusing on ribosomal subunit exchangeability between organisms remain limited. In this study, we show that chimeric ribosomes, composed of Escherichia coli and Bacillus subtilis or E. coli and Geobacillus stearothermophilus subunits, are active for β-galactosidase translation in a highly purified E. coli translation system. Activities of the chimeric ribosomes showed only a modest decrease when using E. coli 30 S subunits, indicating functional conservation of the 50 S subunit between these bacterial species.

  15. Specific radioimmunoassay of HCG and its α and β subunits: methods and results

    International Nuclear Information System (INIS)

    Reuter, A.M.; Schoonbrood, J.; Franchimont, P.

    1976-01-01

    To create antisera that are specific for the radioimmunoassay of HCG and its subunits, the antisera are neutralized by incubation with LH or HCG. For each RIA system the inhibition curves of HCG and its subunits LH, FSH, TSH and STH are obtained. The 125 I labelled hormones HCG, α and β subunits and LH were chromatographed over a Sephadex G 100 column. Serum of menopausal and pregnant women were chromatographed in the same way and the fractions subjected to RIA. HCG and its subunits were determined by RIA in the sera of patients with different kinds of cancer

  16. Thermal aggregation behaviour of soy protein: characteristics of different polypeptides and sub-units.

    Science.gov (United States)

    He, Xiu-Ting; Yuan, De-Bao; Wang, Jin-Mei; Yang, Xiao-Quan

    2016-03-15

    Due to the differences in structure and composition of glycinin and β-conglycinin, they exhibit different characteristics during heat treatment. In present study, the thermal aggregation behaviour of glycinin, β-conglycinin and their isolated sub-units was investigated at pH 7.0. Acidic polypeptides, basic polypeptides, αα' and β sub-units of soy protein were denatured during the isolation process. The degree of aggregation of protein fractions after heat treatment was in the order: denatured basic polypeptides > native glycinin > denatured β sub-unit > native β-conglycinin > denatured acidic polypeptides > denatured αα' sub-units. Glycinin, β-conglycinin, acidic polypeptides and αα'/β sub-units exhibited different changing trends of surface hydrophobicity with increasing temperature. The αα' sub-units showed higher ability to suppress thermal aggregation of basic polypeptides than β sub-units during heat treatment. The β sub-units were shown to form soluble aggregates with glycinin after heating. The interaction mechanism of αα' and β sub-units heated with basic polypeptides was proposed. For the β sub-units-basic polypeptides mixed system, more hydrophobic chains were binding together and buried inside during heat treatment, which resulted in lower surface hydrophobicity. The αα' sub-units-basic polypeptides mixed system was considered to be a stable system with higher surface hydrophobicity after being heated. © 2015 Society of Chemical Industry.

  17. Chaperonin Structure - The Large Multi-Subunit Protein Complex

    Directory of Open Access Journals (Sweden)

    Irena Roterman

    2009-03-01

    Full Text Available The multi sub-unit protein structure representing the chaperonins group is analyzed with respect to its hydrophobicity distribution. The proteins of this group assist protein folding supported by ATP. The specific axial symmetry GroEL structure (two rings of seven units stacked back to back - 524 aa each and the GroES (single ring of seven units - 97 aa each polypeptide chains are analyzed using the hydrophobicity distribution expressed as excess/deficiency all over the molecule to search for structure-to-function relationships. The empirically observed distribution of hydrophobic residues is confronted with the theoretical one representing the idealized hydrophobic core with hydrophilic residues exposure on the surface. The observed discrepancy between these two distributions seems to be aim-oriented, determining the structure-to-function relation. The hydrophobic force field structure generated by the chaperonin capsule is presented. Its possible influence on substrate folding is suggested.

  18. HCG-subunits in choriocarcinoma and in pregnancy

    International Nuclear Information System (INIS)

    Talas, M.; Fingerova, H.; Kudela, M.

    1979-01-01

    Pure HCG-α and HCG-β and anti HCG-β serum were used in the study of HCG-subunits. The following RIA systems were studied: HCG* - anti HCG, HCG* - anti HCG-β, HCG-β* - anti HCG-β, HCG-β* - anti HCG and HCG-α* - anti HCG. HCG-α, HCG-β and the 2nd International Standard of HCG were used as standard preparations. The sensitivity and parallelism of individual inhibition curves were compared. Samples obtained from women with choriocarcinoma and in pregnancy were estimated in RIA systems studied. It was found out that the values of HCG in women treated for choriocarcinoma as obtained in the system HCG* - anti HCG-β were higher than those obtained in the system HCG* - anti HCG. (author)

  19. Determination of hCG-alpha subunit in threatened pregnancy

    International Nuclear Information System (INIS)

    Talas, M.; Pohanka, J.; Fingerova, H.; Janouskova, M.; Krikal, Z.; Prasilova, J.; Zupkova, H.

    1987-01-01

    Radioimmunoassay of the hCG-alpha subunit was made using an antibody anti hCG-alpha serum, highly purified hCG-alpha for 125 I-labelling and the standard hCG-alpha. Sera of healthy pregnant women sampled throughout the whole pregnancies were used to determine x-bar±S.D. of hCG-alpha for 14-day intervals. Included in the study were groups of women with high risk of premature labor, late toxemia of pregnancy, twins and fetal hypotrophy. It was shown that increased hCG-alpha is found in pregnant women in whom signs of late toxemia of pregnancy are combined with high risk of premature labor, or with twin pregnancies, while in those with fetal hypotrophy hCG-alpha is within normal limits. (author). 3 figs., 7 refs

  20. Glycine Receptor α2 Subunit Activation Promotes Cortical Interneuron Migration

    Directory of Open Access Journals (Sweden)

    Ariel Avila

    2013-08-01

    Full Text Available Glycine receptors (GlyRs are detected in the developing CNS before synaptogenesis, but their function remains elusive. This study demonstrates that functional GlyRs are expressed by embryonic cortical interneurons in vivo. Furthermore, genetic disruption of these receptors leads to interneuron migration defects. We discovered that extrasynaptic activation of GlyRs containing the α2 subunit in cortical interneurons by endogenous glycine activates voltage-gated calcium channels and promotes calcium influx, which further modulates actomyosin contractility to fine-tune nuclear translocation during migration. Taken together, our data highlight the molecular events triggered by GlyR α2 activation that control cortical tangential migration during embryogenesis.

  1. The subunit structure of the extracellular hemoglobin of Biomphalaria glabrata

    International Nuclear Information System (INIS)

    Arndt, Marcio H.L.; Naves, Cristiani F.; Xavier, Luciana P.; Santoro, Marcelo M.

    1997-01-01

    Full text. The hemoglobin of Biomphalaria glabrata was purified to homogeneity by a two step purification protocol using a gel filtration column (Superose 6 HR/Pharmacia ) followed by an anion exchange chromatography (MONO-Q Sepharose/Pharmacia). The dissociation products were analysed by a 5 - 15 % Polyacrylamide gel electrophoresis containing Sodium Dodecyl Sulfate (SDS-PAGE) giving a band of 270 K Daltons and a band of 180 K Daltons after reduction with β-mercaptoethanol. The same profile was obtained in a 3.5 % Agarose gel electrophoresis containing SDS (SDS-AGE) showing additional bands of higher molecular weight. These bands were proposed to be monomers, dimers and trimers and, after reduction in a Bidimensional SDS-AGE, the proposed monomers and dimers were decomposed in two and four bands that were interpreted as 1 - 4 chains. The hemoglobin was digested by four different proteases ( Thrombin, Trypsin, Chymotrypsin and Subtilisin ) showing several equivalent fragments with molecular weights multiples of its minimum molecular weight ( 17.7 K Daltons). The circular dichroism spectrum of the protein showed a characteristic high α-helix content. We proposed that this hemoglobin is a pentamer of approx. 360 K Daltons subunits each formed by two 180 K Daltons chains linked in pairs by disulfide bridges and each of these chains comprises ten Heme binding domains. These data were compared to other Planorbidae extracellular hemoglobins. Up to now, the quaternary structure of this hemoglobin (shape and disposition of the subunits) is unknown. It is intended to elucidate its structure by Small Angle X-Ray Scattering in Brazilian National Laboratory of Synchrotron Light (LNLS). (author)

  2. Differential regulation of thyrotropin subunit apoprotein and carbohydrate biosynthesis by thyroid hormone

    Energy Technology Data Exchange (ETDEWEB)

    Taylor, T.; Weintraub, B.D.

    1985-04-01

    The regulation of TSH apoprotein and carbohydrate biosynthesis by thyroid hormone was studied by incubating pituitaries from normal and hypothyroid (3 weeks post-thyroidectomy) rats in medium containing (/sup 14/C)alanine and (/sup 3/H) glucosamine. After 6 h, samples were sequentially treated with anti-TSH beta to precipitate TSH and free TSH beta, anti-LH beta to clear the sample of LH and free LH beta, then anti-LH alpha to precipitate free alpha-subunit. Total proteins were acid precipitated. All precipitates were subjected to electrophoresis on sodium dodecyl sulfate-polyacrylamide gels, which were then sliced and assayed by scintillation spectrometry. In hypothyroid pituitaries plus medium, (/sup 14/C)alanine incorporation in combined and free beta-subunits was 26 times normal and considerably greater than the 3.4-fold increase seen in total protein; combined and free alpha-subunits showed no specific increase in apoprotein synthesis. (/sup 3/H)Glucosamine incorporation in combined alpha- and beta-subunits in hypothyroid samples was 13 and 21 times normal, respectively, and was greater than the 1.9-fold increase in total protein; free alpha-subunit showed no specific increase in carbohydrate synthesis. The glucosamine to alanine ratio, reflecting relative glycosylation of newly synthesized molecules, was increased in hypothyroidism for combined alpha-subunits, but not for combined beta-subunits, free alpha-subunits, or total proteins. In summary, short term hypothyroidism selectively stimulated TSH beta apoprotein synthesis and carbohydrate synthesis of combined alpha- and beta-subunits. Hypothyroidism also increased the relative glycosylation of combined alpha-subunit. Thus, thyroid hormone deficiency appears to alter the rate-limiting step in TSH assembly (i.e. beta-subunit synthesis) as well as the carbohydrate structure of TSH, which may play important roles in its biological function.

  3. Copolymer semiconductors comprising thiazolothiazole or benzobisthiazole, or benzobisoxazole electron acceptor subunits, and electron donor subunits, and their uses in transistors and solar cells

    Science.gov (United States)

    Jenekhe, Samson A; Subramaniyan, Selvam; Ahmed, Eilaf; Xin, Hao; Kim, Felix Sunjoo

    2014-10-28

    The inventions disclosed, described, and/or claimed herein relate to copolymers comprising copolymers comprising electron accepting A subunits that comprise thiazolothiazole, benzobisthiazole, or benzobisoxazoles rings, and electron donating subunits that comprise certain heterocyclic groups. The copolymers are useful for manufacturing organic electronic devices, including transistors and solar cells. The invention also relates to certain synthetic precursors of the copolymers. Methods for making the copolymers and the derivative electronic devices are also described.

  4. Roles of the β subunit hinge domain in ATP synthase F1 sector: Hydrophobic network formed by introduced βPhe174 inhibits subunit rotation

    International Nuclear Information System (INIS)

    Nakanishi-Matsui, Mayumi; Kashiwagi, Sachiko; Kojima, Masaki; Nonaka, Takamasa; Futai, Masamitsu

    2010-01-01

    The ATP synthase β subunit hinge domain (βPhe148 ∼ βGly186, P-loop/α-helixB/loop/β-sheet4, Escherichia coli residue numbering) dramatically changes in conformation upon nucleotide binding. We previously reported that F 1 with the βSer174 to Phe mutation in the domain lowered the γ subunit rotation speed, and thus decreased the ATPase activity [M. Nakanishi-Matsui, S. Kashiwagi, T. Ubukata, A. Iwamoto-Kihara, Y. Wada, M. Futai, Rotational catalysis of Escherichia coli ATP synthase F 1 sector. Stochastic fluctuation and a key domain of the β subunit, J. Biol. Chem. 282 (2007) 20698-20704.]. Homology modeling indicates that the amino acid replacement induces a hydrophobic network, in which the βMet159, βIle163, and βAla167 residues of the β subunit are involved together with the mutant βPhe174. The network is expected to stabilize the conformation of β DP (nucleotide-bound form of the β subunit), resulting in increased activation energy for transition to β E (empty β subunit). The modeling further predicts that replacement of βMet159 with Ala or Ile weakens the hydrophobic network. As expected, these two mutations experimentally suppressed the ATPase activities as well as subunit rotation of βS174F. Furthermore, the rotation rate decreased with the increase of the strength in the hydrophobic network. These results indicate that the smooth conformational change of the β subunit hinge domain is pertinent for the rotational catalysis.

  5. Influence of ancillary ligands on preferential geometry and biomimetic catalytic activity in manganese(III)-catecholate systems: A combined experimental and theoretical study.

    Science.gov (United States)

    Jana, Narayan Ch; Brandão, Paula; Bauzá, Antonio; Frontera, Antonio; Panja, Anangamohan

    2017-11-01

    The present report describes the synthesis and structural characterizations of six new manganese(III) complexes with redox-active tetrachlorocatecholate ligand in the presence of different ancillary ligands (pyridines and imidazole). X-ray crystal structure analysis reveals that the geometry of manganese(III) centres in 1 and 2 is essentially square pyramidal, while it is discrete octahedron in compounds 3-6. These preferential structural diversities in these systems have been critically analysed by theoretical calculations. Remarkably, the characterization of both π⋯π stacking interactions and MnMn bonds in the supramolecular dimeric aggregates in the solid state in 1 and 2 by means of the Bader's theory of "atoms in molecules" (AIM) is quite interesting as that nicely corroborates the experimental fact. All the complexes are active toward the phenoxazinone synthase like activity and the detailed kinetic analysis was performed to get better insight into their catalytic efficiency. Electrochemical property of these complexes as well as different donor property of the ancillary ligands clearly establish that the ease of reduction of the metal centre i.e., the catalytic ability is favoured when the metal centre is bonded to the electron deficient pyridyl systems. EPR spectroscopy and theoretical study are further helpful to get insight into origin of the catalytic activity in these compounds. The present report overall highlights that tuning of the geometry and catalytic activity of manganese(III) complexes with tetrachlorocatecholate ligand can be attained by the introduction of different substitutions in ancillary pyridine ligands. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Smart microgrid hierarchical frequency control ancillary service provision based on virtual inertia concept: An integrated demand response and droop controlled distributed generation framework

    International Nuclear Information System (INIS)

    Rezaei, Navid; Kalantar, Mohsen

    2015-01-01

    Highlights: • Detailed formulation of the microgrid static and dynamic securities based on droop control and virtual inertia concepts. • Constructing a novel objective function using frequency excursion and rate of change of frequency profiles. • Ensuring the microgrid security subject to the microgrid economic and environmental policies. • Coordinated management of demand response and droop controlled distributed generation resources. • Precise scheduling of day-ahead hierarchical frequency control ancillary services using a scenario based stochastic programming. - Abstract: Low inertia stack, high penetration levels of renewable energy source and great ratio of power deviations in a small power delivery system put microgrid frequency at risk of instability. On the basis of the close coupling between the microgrid frequency and system security requirements, procurement of adequate ancillary services from cost-effective and environmental friendly resources is a great challenge requests an efficient energy management system. Motivated by this need, this paper presents a novel energy management system that is aimed to coordinately manage the demand response and distributed generation resources. The proposed approach is carried out by constructing a hierarchical frequency control structure in which the frequency dependent control functions of the microgrid components are modeled comprehensively. On the basis of the derived modeling, both the static and dynamic frequency securities of an islanded microgrid are provided in primary and secondary control levels. Besides, to cope with the low inertia stack of islanded microgrids, novel virtual inertia concept is devised based on the precise modeling of droop controlled distributed generation resources. The proposed approach is applied to typical test microgrid. Energy and hierarchical reserve resource are scheduled precisely using a scenario-based stochastic programming methodology. Moreover, analyzing the

  7. The Use of Ancillary Stains in the Diagnosis of Barrett Esophagus and Barrett Esophagus-associated Dysplasia: Recommendations From the Rodger C. Haggitt Gastrointestinal Pathology Society.

    Science.gov (United States)

    Srivastava, Amitabh; Appelman, Henry; Goldsmith, Jeffrey D; Davison, Jon M; Hart, John; Krasinskas, Alyssa M

    2017-05-01

    Barrett esophagus (BE) is a known risk factor for the development of esophageal adenocarcinoma. Pathologists play a critical role in confirming the diagnosis of BE and BE-associated dysplasia. As these diagnoses are not always straightforward on routine hematoxylin and eosin-stained slides, numerous ancillary stains have been used in an attempt to help pathologists confirm the diagnosis. On the basis of an in-depth review of the literature, the Rodger C. Haggitt Gastrointestinal Pathology Society provides recommendations regarding the use of ancillary stains in the diagnosis of BE and BE-associated dysplasia. Because goblet cells are almost always identifiable on routine hematoxylin and eosin-stained sections, there is insufficient evidence to justify reflexive use of Alcian blue (at pH 2.5) and/or periodic-acid Schiff stains on all esophageal biopsies to diagnose BE. In addition, the use of mucin glycoprotein immunostains and markers of intestinal phenotype (CDX2, Das-1, villin, Hep Par 1, and SOX9) are not indicated to aid in the diagnosis of BE at this time. A diagnosis of dysplasia in BE remains a morphologic diagnosis, and hence, ancillary stains are not recommended for diagnosing dysplasia. Although p53 is a promising marker for identifying high-risk BE patients, it is not recommended for routine use at present; additional studies are needed to address questions regarding case selection, interpretation, integration with morphologic diagnosis, and impact on clinical outcome. We hope that this review and our recommendations will provide helpful information to pathologists, gastroenterologists, and others involved in the evaluation of patients with BE and BE-associated dysplasia.

  8. Phosphorescent cationic iridium(iii) complexes bearing a nonconjugated six-membered chelating ancillary ligand: a strategy for tuning the emission towards the blue.

    Science.gov (United States)

    Hierlinger, Claus; Cordes, David B; Slawin, Alexandra M Z; Jacquemin, Denis; Guerchais, Véronique; Zysman-Colman, Eli

    2018-03-15

    This study concerns an assessment of the impact of the interruption of the electronic crosstalk between the pyridine rings in the ancillary ligand on the photoluminescence properties of the corresponding iridium(iii) complexes. Two new cationic Ir(iii) complexes, [Ir(dFmesppy) 2 (pmdp)]PF 6 , 1, and [Ir(mesppy) 2 (pmdp)]PF 6 , 2, [where dFmesppy is 2-(2,4-difluorophenyl)-4-mesitylpyridinato, mesppy is 4-mesityl-2-phenylpyridinato and pmdp is 2,2'-(phenylmethine)dipyridine, L1] possessing sterically congested cyclometalating ligands combined with the nonconjugated diimine ancillary N^N ligand are reported and their solution-state and thin film photophysical properties analyzed by both experimental and theoretical methods. The crystal structure of 1 confirms the formation of a six-membered chelate ring by the N^N ligand and illustrates the pseudo-axial configuration of the phenyl substituents. Upon photoexcitation in acetonitrile, both complexes exhibit a ligand-centered emission profile in the blue-green region of the visible spectrum. A significant blue-shift is observed in solution at room temperature compared to the analogous reference Ir(iii) complexes (R1 and R2) bearing 4,4'-di-tert-butyl-2,2'-bipyridine (dtBubpy) as the N^N ligand. The computational investigation demonstrates that the HOMO is mainly centered on the metal and on both cyclometalating aryl rings of the C^N ligands, whereas the LUMO is principally localized on the pyridyl rings of the C^N ligands. The photoluminescence quantum yield is reduced compared to the reference complexes, a probable consequence of the greater flexibility of the ancillary ligand.

  9. Intravital imaging reveals new ancillary mechanisms co-opted by cancer cells to drive tumor progression [version 1; referees: 2 approved

    Directory of Open Access Journals (Sweden)

    Claire Vennin

    2016-05-01

    Full Text Available Intravital imaging is providing new insights into the dynamics of tumor progression in native tissues and has started to reveal the layers of complexity found in cancer. Recent advances in intravital imaging have allowed us to look deeper into cancer behavior and to dissect the interactions between tumor cells and the ancillary host niche that promote cancer development. In this review, we provide an insight into the latest advances in cancer biology achieved by intravital imaging, focusing on recently discovered mechanisms by which tumor cells manipulate normal tissue to facilitate disease progression.

  10. Peer-to-Peer Consultations: Ancillary Services Peer Exchange with India: Experience from South Africa, Europe & the United States (Fact Sheet)

    Energy Technology Data Exchange (ETDEWEB)

    2014-05-01

    In support of national and subnational decision makers, the 21st Century Power Partnership regularly works with country partners to organize peer-to-peer consultations on critical issues. In March 2014, 21CPP collaborated with the Regulatory Assistance Project - India to host two peer-to-peer exchanges among experts from India, South Africa, Europe, and the United States to discuss the provision of ancillary services, particularly in the context of added variability and uncertainty from renewable energy. This factsheet provides a high level summary of the peer-to-peer consultation.

  11. [Nature of the heterogeneity of the 30S ribosomal subunits in vitro. II. Two types of inactivation of the 30S subunits of Escherichia coli ribosomes].

    Science.gov (United States)

    Peshin, N N; Kirillov, S V

    1979-01-01

    The influence of concentration of monovalent cations on the binding constant of Phe-tRNAPhe to 30S.poly(U) complex was studied. Two types of inactivation of the 30S subunits by ammonium ions at the low magnesium concentration (1 mM) were found. The first type of inactivation was observed at high concentrations of NH4+ ions (from 0.5 to 1.5 M) and due to the dissociation of ribosomal proteins from 30S subunits. This inactivation only decreased the binding constant of Phe-tRNAPhe to 30S.poly(U) complex up to 50 times but all 30S subunits were equally achieved in Phe-tRNAPhe binding. This type of inactivation was reversible, addition of S-proteins restored the association constant to the original value. At low concentration of NH4+ ions (below 100 mM) about half of the 30S subunits is irreversibly inactivated (the binding constant of Phe-tRNAPhe decreased below detectable level) probably as a result of conformational changes in ribosomal RNA. Both types of inactivation of the 30S subunits can take place during the preparation of isolated subunits of ribosomes.

  12. Differential antibiotic sensitivity determined by the large ribosomal subunit in thermophilic archaea.

    OpenAIRE

    Ruggero, D; Londei, P

    1996-01-01

    Hybrid ribosomes obtained by mixing the ribosomal subunits of the extremely thermophilic archaea Sulfolobus solfataricus and Desulfurococcus mobilis were tested for their sensitivity to selected antibiotics. It is shown that structural differences in the large ribosomal subunits determine qualitatively and quantitatively the patterns of response to alpha-sarcin and paromomycin in these species.

  13. A-Raf kinase is a new interacting partner of protein kinase CK2 beta subunit

    DEFF Research Database (Denmark)

    Boldyreff, B; Issinger, O G

    1997-01-01

    In a search for protein kinase CK2 beta subunit binding proteins using the two-hybrid system, more than 1000 positive clones were isolated. Beside clones for the alpha' and beta subunit of CK2, there were clones coding for a so far unknown protein, whose partial cDNA sequence was already deposited...

  14. Isoforms of human cytochrome-c oxidase. Subunit composition and steady-state kinetic properties

    NARCIS (Netherlands)

    van Kuilenburg, A. B.; Dekker, H. L.; van den Bogert, C.; Nieboer, P.; van Gelder, B. F.; Muijsers, A. O.

    1991-01-01

    The subunit pattern and the steady-state kinetics of cytochrome-c oxidase from human heart, muscle, kidney and liver were investigated. Polyacrylamide gel electrophoresis of immunopurified cytochrome-c oxidase preparations suggest that isoforms of subunit VIa exist, which show differences in

  15. Regulated appearance of NMDA receptor subunits and channel functions during in vitro neuronal differentiation

    NARCIS (Netherlands)

    Jelitai, Márta; Schlett, Katalin; Varju, Patrícia; Eisel, Ulrich; Madarász, Emília

    The schedule of NMDA receptor subunit expression and the appearance of functional NMDA-gated ion channels were investigated during the retinoic acid (RA) induced neuronal differentiation of NE-4C, a p53-deficient mouse neuroectodermal progenitor cell line. NR2A. NR2B, and NR2D subunit transcripts

  16. Human heart cytochrome c oxidase subunit VIII. Purification and determination of the complete amino acid sequence

    NARCIS (Netherlands)

    van Kuilenburg, A. B.; Muijsers, A. O.; Demol, H.; Dekker, H. L.; van Beeumen, J. J.

    1988-01-01

    Subunit VIII was purified from a preparation of the human heart cytochrome c oxidase and its complete amino acid sequence was determined. The sequence proved to be much more related to that of the bovine liver oxidase subunit VIII than to that found in bovine heart. Our finding of a 'liver-type'

  17. Lack of NMDA receptor subunit exchange alters Purkinje cell dendritic morphology in cerebellar slice cultures

    NARCIS (Netherlands)

    Metzger, F; Pieri, [No Value; Eisel, ULM; Pieri, Isabelle

    2005-01-01

    Early postnatal developmental changes in N-methyl-D-aspartate (NMDA) receptor (NR) subunits regulate cerebellar granule cell maturation and potentially Purkinje cell development. We therefore investigated Purkinje cell morphology in slice cultures from mice with genetic subunit exchange from NR2C to

  18. Specific Inhibition of Herpes Simplex Virus DNA Polymerase by Helical Peptides Corresponding to the Subunit Interface

    Science.gov (United States)

    Digard, Paul; Williams, Kevin P.; Hensley, Preston; Brooks, Ian S.; Dahl, Charles E.; Coen, Donald M.

    1995-02-01

    The herpes simplex virus DNA polymerase consists of two subunits-a catalytic subunit and an accessory subunit, UL42, that increases processivity. Mutations affecting the extreme C terminus of the catalytic subunit specifically disrupt subunit interactions and ablate virus replication, suggesting that new antiviral drugs could be rationally designed to interfere with polymerase heterodimerization. To aid design, we performed circular dichroism (CD) spectroscopy and analytical ultracentrifugation studies, which revealed that a 36-residue peptide corresponding to the C terminus of the catalytic subunit folds into a monomeric structure with partial α-helical character. CD studies of shorter peptides were consistent with a model where two separate regions of α-helix interact to form a hairpin-like structure. The 36-residue peptide and a shorter peptide corresponding to the C-terminal 18 residues blocked UL42-dependent long-chain DNA synthesis at concentrations that had no effect on synthesis by the catalytic subunit alone or by calf thymus DNA polymerase δ and its processivity factor. These peptides, therefore, represent a class of specific inhibitors of herpes simplex virus DNA polymerase that act by blocking accessory-subunit-dependent synthesis. These peptides or their structures may form the basis for the synthesis of clinically effective drugs.

  19. Expression of the scaffolding subunit A of protein phosphatase 2A during rat testicular development

    NARCIS (Netherlands)

    van den Ham, R.; van Dissel-Emiliani, F. M. F.; van Pelt, A. M. M.

    2003-01-01

    Previously, we found that the poly(A)+ RNA of the scaffolding subunit A (alpha isoform) of protein phosphatase 2A (PP2A-Aalpha) was clearly expressed by fetal gonocytes but weakly expressed by adult single (As), paired (Apr), and aligned (Aal) A spermatogonia. The scaffolding subunit A of PP2A

  20. Regulation of KV channel voltage-dependent activation by transmembrane β subunits

    Directory of Open Access Journals (Sweden)

    Xiaohui eSun

    2012-04-01

    Full Text Available Voltage-activated K+ (KV channels are important for shaping action potentials and maintaining resting membrane potential in excitable cells. KV channels contain a central pore-gate domain (PGD surrounded by four voltage-sensing domains (VSD. The VSDs will change conformation in response to alterations of the membrane potential thereby inducing the opening of the PGD. Many KV channels are heteromeric protein complexes containing auxiliary β subunits. These β subunits modulate channel expression and activity to increase functional diversity and render tissue specific phenotypes. This review focuses on the KV β subunits that contain transmembrane (TM segments including the KCNE family and the β subunits of large conductance, Ca2+- and voltage-activated K+ (BK channels. These TM β subunits affect the voltage-dependent activation of KV α subunits. Experimental and computational studies have described the structural location of these β subunits in the channel complexes and the biophysical effects on VSD activation, PGD opening and VSD-PGD coupling. These results reveal some common characteristics and mechanistic insights into KV channel modulation by TM β subunits.

  1. Visualization of subunit interactions and ternary complexes of protein phosphatase 2A in mammalian cells.

    Directory of Open Access Journals (Sweden)

    Shu-Ting Mo

    Full Text Available Protein phosphatase 2A (PP2A is a ubiquitous phospho-serine/threonine phosphatase that controls many diverse cellular functions. The predominant form of PP2A is a heterotrimeric holoenzyme consisting of a scaffolding A subunit, a variable regulatory B subunit, and a catalytic C subunit. The C subunit also associates with other interacting partners, such as α4, to form non-canonical PP2A complexes. We report visualization of PP2A complexes in mammalian cells. Bimolecular fluorescence complementation (BiFC analysis of PP2A subunit interactions demonstrates that the B subunit plays a key role in directing the subcellular localization of PP2A, and confirms that the A subunit functions as a scaffold in recruiting the B and C subunits to form a heterotrimeric holoenzyme. BiFC analysis also reveals that α4 promotes formation of the AC core dimer. Furthermore, we demonstrate visualization of specific ABC holoenzymes in cells by combining BiFC and fluorescence resonance energy transfer (BiFC-FRET. Our studies not only provide direct imaging data to support previous biochemical observations on PP2A complexes, but also offer a promising approach for studying the spatiotemporal distribution of individual PP2A complexes in cells.

  2. The A and B variants of the alpha 3 integrin subunit: tissue distribution and functional characterization

    NARCIS (Netherlands)

    de Melker, A. A.; Sterk, L. M.; Delwel, G. O.; Fles, D. L.; Daams, H.; Weening, J. J.; Sonnenberg, A.

    1997-01-01

    The alpha subunits of the laminin-binding integrins alpha 3 beta 1, alpha 6 beta 1, and alpha 7 beta 1 have homologous sequences and are similar in structure. Two cytoplasmic variants, A and B, have been identified for each of these alpha subunits, although the alpha 3B splice variant has been

  3. A-Raf kinase is a new interacting partner of protein kinase CK2 beta subunit

    DEFF Research Database (Denmark)

    Boldyreff, B; Issinger, O G

    1997-01-01

    In a search for protein kinase CK2 beta subunit binding proteins using the two-hybrid system, more than 1000 positive clones were isolated. Beside clones for the alpha' and beta subunit of CK2, there were clones coding for a so far unknown protein, whose partial cDNA sequence was already deposite...

  4. Purification of the alpha and beta subunits of phosphorylase kinase for structural studies

    International Nuclear Information System (INIS)

    Sotiroudis, T.G.; Heilmeyer, L.M.G. Jr.; Crabb, J.W.

    1987-01-01

    Structural analysis of the alpha (Mr, 132,000) and beta (Mr, 113,000) subunits of phosphorylase kinase may provide clues to their yet unknown functions however purification remains problematic. Preparative RP-HPLC procedures yield inconveniently large, dilute solutions and concentration steps are required prior to subunit modification and fragmentation. Concentration of the β subunit usually results in significant losses due to insolubility. Using preparative SDS-polyacrylamide gel electrophoresis, they have purified the α, 7 , and β subunits from rabbit muscle phosphorylase kinase in a soluble and concentrated form suitable for structural studies. Phosphorylase kinase labelled with fluorescein isothiocyanate in the α and α' subunits and fully 14 C-S-carboxymethylated was fractionated on a 5% acrylamide Laemmli slab gel. The subunit bands were visualized by fluorescence and by SDS precipitation then excised and electroeluted in the presence of SDS using an ELUTRAP device. From 4.5 mg of enzyme applied to a 4.5 mm thick gel about 70% of the α subunit and about 90% of the β subunit were typically recovered in less than 1 ml with overnight elution

  5. Facilitation by the beta2a subunit of pore openings in cardiac Ca2+ channels.

    Science.gov (United States)

    Costantin, J; Noceti, F; Qin, N; Wei, X; Birnbaumer, L; Stefani, E

    1998-02-15

    1. Single channel recordings were performed on the cardiac calcium channel (alpha1C) in order to study the effect of coexpression of the accessory beta2a subunit. On-cell patch clamp recordings were performed after expression of these channels in Xenopus oocytes. 2. The alpha1C subunit, when expressed alone, had similar single channel properties to native cardiac channels. Slow transitions between low and high open probability (Po) gating modes were found as well as fast gating transitions between the open and closed states. 3. Coexpression of the beta2a subunit caused changes in the fast gating during high Po mode. In this mode, open time distributions reveal at least three open states and the beta2a subunit favours the occupancy of the longest, 10-15 ms open state. No effect of the beta2a subunit was found when the channel was gating in the low Po mode. 4. Slow gating transitions were also affected by the beta2a subunit. The high Po mode was maintained for the duration of the depolarizing pulse in the presence of the beta2a subunit; while the alpha1C channel when expressed alone, frequently switched into and out of the high Po mode during the course of a sweep. 5. The beta2a subunit also affected mode switching that occurred between sweeps. Runs analysis revealed that the alpha1C subunit has a tendency toward non-random mode switching. The beta2a subunit increased this tendency. A chi2 analysis of contingency tables indicated that the beta2a subunit caused the alpha1C channel to gain 'intrinsic memory', meaning that the mode of a given sweep can be non-independent of the mode of the previous sweep. 6. We conclude that the beta2a subunit causes changes to the alpha1C channel in both its fast and slow gating behaviour. The beta2a subunit alters fast gating by facilitating movement of the channel into an existing open state. Additionally, the beta2a subunit decreases the slow switching between low and high Po modes.

  6. Pattern of secretion of immunoreactive inhibin/activin subunits by avian granulosa cells.

    Science.gov (United States)

    Johnson, P A; Brooks, C F; Davis, A J

    2005-05-01

    The messenger RNA expression for the inhibin/activin subunits in the granulosa layer of avian follicles of different developmental stages has previously been reported. In the present study, we examined the pattern of secretion of these protein subunits from cultured granulosa cells (GC) of avian follicles of defined maturity. Laying hens were euthanized and the F1, F2, F3, F4, small yellow follicles (SYF; 6-10 mm) and large white follicles (LWF; 3-5 mm) were removed. GC were isolated from the follicles, plated by size at a density of 6.25 x 10(5)cells per well (3 wells per follicle size) and cultured for 48 h in medium 199 with 5% FBS, antibiotics, and 1.0 microg/ml bovine insulin. After 48 h, the cultures were terminated and the media were saved (n = 6 replications). Proteins were precipitated from media, reconstituted for electrophoresis (SDS-PAGE), and analyzed by Western blot. Progesterone was also measured in the medium. For detection of the inhibin alpha-subunit, a rabbit antibody against the chicken inhibin alpha-subunit (1-26 aa) was used. The betaA-subunit was detected with rabbit anti-betaA-subunit (81-113 aa) and the betaB-subunit was detected with rabbit anti-betaB-subunit (80-112 aa). Under reduced conditions, GC from the larger follicle sizes (F1-F4) secreted the most (p subunit compared to smaller follicle sizes. Under non-reduced conditions, a band at approximately 32 kDa was detected by both the alpha-subunit antibody and by the betaA-subunit antibody in media from GC of the F1-F4 follicles, suggesting secretion of intact inhibin A. Immunoreactive alpha-subunit and betaB-subunit were detected under reduced conditions in media from the GC of the SYF, suggesting that this follicle population may secrete intact inhibin B. In addition, under non-reduced conditions, cells from the SYF secreted the greatest amount of intact inhibin B (p subunits. Progesterone concentration in the media from the F1 follicle was greatest and was decreased in media from

  7. Isolation and characterization of recombinant human casein kinase II subunits alpha and beta from bacteria

    DEFF Research Database (Denmark)

    Grankowski, N; Boldyreff, B; Issinger, O G

    1991-01-01

    cDNA encoding the casein kinase II (CKII) subunits alpha and beta of human origin were expressed in Escherichia coli using expression vector pT7-7. Significant expression was obtained with E. coli BL21(DE3). The CKII subunits accounted for approximately 30% of the bacterial protein; however, most...... of the expressed proteins were produced in an insoluble form. The recombinant CKII alpha subunit was purified by DEAE-cellulose chromatography, followed by phosphocellulose and heparin-agarose chromatography. The recombinant CKII beta subunit was extracted from the insoluble pellet and purified in a single step...... at a 1:1 ratio of both subunits. Using a synthetic peptide (RRRDDDSDDD) as a substrate, the maximum protein kinase stimulation observed was fourfold under the conditions used. The Km of the reconstituted enzyme for the synthetic peptide (80 microM) was comparable to the mammalian enzyme (40-60 micro...

  8. Fluorescence resonance energy transfer analysis of subunit assembly of the ASIC channel.

    Science.gov (United States)

    Gao, Ying; Liu, Shuang-Shuang; Qiu, Shuang; Cheng, Wei; Zheng, Jie; Luo, Jian-Hong

    2007-07-20

    Acid-sensing ion channels (ASICs) are believed to be homo- or heteromeric complexes, which have been verified by classical methods such as co-immunoprecipitation or electrophysiological assays. However, the exact subunit combinations of ASICs in living cells have not been established yet. Here, we apply assays based on fluorescence resonance energy transfer (FRET) between GFP color mutants CFP and YFP to investigate ASIC assembly directly in living cells. Homomerization as well as heteromerization of different combinations of ASIC subunits were found. In addition, our results suggest the formation of heteromeric 1a/2a channels of stoichiometry consisting of at least two 1a subunits and two 2a subunits. Similar stoichiometry was observed from heteromeric 1a/2b and 2a/2b channels. Our results imply that these heteromeric ASIC channels contain at least four subunits.

  9. Persistence of the mitochondrial permeability transition in the absence of subunit c of human ATP synthase.

    Science.gov (United States)

    He, Jiuya; Ford, Holly C; Carroll, Joe; Ding, Shujing; Fearnley, Ian M; Walker, John E

    2017-03-28

    The permeability transition in human mitochondria refers to the opening of a nonspecific channel, known as the permeability transition pore (PTP), in the inner membrane. Opening can be triggered by calcium ions, leading to swelling of the organelle, disruption of the inner membrane, and ATP synthesis, followed by cell death. Recent proposals suggest that the pore is associated with the ATP synthase complex and specifically with the ring of c-subunits that constitute the membrane domain of the enzyme's rotor. The c-subunit is produced from three nuclear genes, ATP5G1 , ATP5G2 , and ATP5G3 , encoding identical copies of the mature protein with different mitochondrial-targeting sequences that are removed during their import into the organelle. To investigate the involvement of the c-subunit in the PTP, we generated a clonal cell, HAP1-A12, from near-haploid human cells, in which ATP5G1 , ATP5G2 , and ATP5G3 were disrupted. The HAP1-A12 cells are incapable of producing the c-subunit, but they preserve the characteristic properties of the PTP. Therefore, the c-subunit does not provide the PTP. The mitochondria in HAP1-A12 cells assemble a vestigial ATP synthase, with intact F 1 -catalytic and peripheral stalk domains and the supernumerary subunits e, f, and g, but lacking membrane subunits ATP6 and ATP8. The same vestigial complex plus associated c-subunits was characterized from human 143B ρ 0 cells, which cannot make the subunits ATP6 and ATP8, but retain the PTP. Therefore, none of the membrane subunits of the ATP synthase that are involved directly in transmembrane proton translocation is involved in forming the PTP.

  10. Submitochondrial distributions and stabilities of subunits 4, 5, and 6 of yeast cytochrome oxidase in assembly defective mutants.

    Science.gov (United States)

    Glerum, D M; Tzagoloff, A

    1997-08-04

    The concentration and submitochondrial distribution of the subunit polypeptides of cytochrome oxidase have been studied in wild type yeast and in different mutants impaired in assembly of this respiratory complex. All the subunit polypeptides of the enzyme are associated with mitochondrial membranes of wild type cells, except for a small fraction of subunits 4 and 6 that is recovered in the soluble protein fraction of mitochondria. Cytochrome oxidase mutants consistently display a severe reduction in the steady-state concentration of subunit 1 due to its increased turnover. As a consequence, most of subunit 4, which normally is associated with subunit 1, is found in the soluble fraction. A similar shift from membrane-bound to soluble subunit 6 is seen in mutants blocked in expression of subunit 5a. In contrast, null mutations in COX6 coding for subunit 6 promote loss of subunit 5a. The absence of subunit 5a in the cox6 mutant is the result of proteolytic degradation rather than regulation of its expression by subunit 6. The possible role of the ATP-dependent proteases Rca1p and Afg3p in proteolysis of subunits 1 and 5a has been assessed in strains with combined mutations in COX6, RCA1, and/or AFG3. Immunochemical assays indicate that another protease(s) must be responsible for most of the proteolytic loss of these proteins.

  11. Vehicle Ancillary Load Reduction Project Close-Out Report: An Overview of the Task and a Compilation of the Research Results

    Energy Technology Data Exchange (ETDEWEB)

    Rugh, J.; Farrington, R.

    2008-01-01

    The amount of fuel used for climate control in U.S. vehicles reduces the fuel economy of more than 200 million light-duty conventional vehicles and thus affects U.S. energy security. Researchers at the DOE National Renewable Energy Laboratory estimated that the United States consumes about 7 billion gallons of fuel per year for air-conditioning (A/C) light-duty vehicles. Using a variety of tools, NREL researchers developed innovative techniques and technologies to reduce the amount of fuel needed for these vehicles' ancillary loads. For example, they found that the A/C cooling capacity of 5.7 kW in a Cadillac STS could be reduced by 30% while maintaining a cooldown performance of 30 minutes. A simulation showed that reducing the A/C load by 30% decreased A/C fuel consumption by 26%. Other simulations supported the great potential for improving fuel economy by using new technologies and techniques developed to reduce ancillary loads.

  12. Mesenteric vascular occlusion: Comparison of ancillary CT findings between arterial and venous occlusions and independent CT findings suggesting life-threatening events

    Energy Technology Data Exchange (ETDEWEB)

    Wong, Yon Cheong; Wu, Cheng Hsien; Wang, Li Jen; Chen, Huan Wu; Lin, Being Chuan; Huang, Chen Chih [Chang Gung Memorial Hospital, Chang Gung University, Taoyuan (China)

    2013-01-15

    To compare the ancillary CT findings between superior mesenteric artery thromboembolism (SMAT) and superior mesenteric vein thrombosis (SMVT), and to determine the independent CT findings of life-threatening mesenteric occlusion. Our study was approved by the institution review board. We included 43 patients (21 SMAT and 22 SMVT between 1999 and 2008) of their median age of 60.0 years, and retrospectively analyzed their CT scans. Medical records were reviewed for demographics, management, surgical pathology diagnosis, and outcome. We compared CT findings between SMAT and SMVT groups. Multivariate analysis was conducted to determine the independent CT findings of life-threatening mesenteric occlusion. Of 43 patients, 24 had life-threatening mesenteric occlusion. Death related to mesenteric occlusion was 32.6%. A thick bowel wall (p < 0.001), mesenteric edema (p < 0.001), and ascites (p = 0.009) were more frequently associated with SMVT, whereas diminished bowel enhancement (p = 0.003) and paralytic ileus (p = 0.039) were more frequent in SMAT. Diminished bowel enhancement (OR = 20; p = 0.007) and paralytic ileus (OR = 16; p = 0.033) were independent findings suggesting life-threatening mesenteric occlusion. The ancillary CT findings occur with different frequencies in SMAT and SMVT. However, the independent findings indicating life-threatening mesenteric occlusion are diminished bowel wall enhancement and paralytic ileus.

  13. Mesenteric vascular occlusion: Comparison of ancillary CT findings between arterial and venous occlusions and independent CT findings suggesting life-threatening events

    International Nuclear Information System (INIS)

    Wong, Yon Cheong; Wu, Cheng Hsien; Wang, Li Jen; Chen, Huan Wu; Lin, Being Chuan; Huang, Chen Chih

    2013-01-01

    To compare the ancillary CT findings between superior mesenteric artery thromboembolism (SMAT) and superior mesenteric vein thrombosis (SMVT), and to determine the independent CT findings of life-threatening mesenteric occlusion. Our study was approved by the institution review board. We included 43 patients (21 SMAT and 22 SMVT between 1999 and 2008) of their median age of 60.0 years, and retrospectively analyzed their CT scans. Medical records were reviewed for demographics, management, surgical pathology diagnosis, and outcome. We compared CT findings between SMAT and SMVT groups. Multivariate analysis was conducted to determine the independent CT findings of life-threatening mesenteric occlusion. Of 43 patients, 24 had life-threatening mesenteric occlusion. Death related to mesenteric occlusion was 32.6%. A thick bowel wall (p < 0.001), mesenteric edema (p < 0.001), and ascites (p = 0.009) were more frequently associated with SMVT, whereas diminished bowel enhancement (p = 0.003) and paralytic ileus (p = 0.039) were more frequent in SMAT. Diminished bowel enhancement (OR = 20; p = 0.007) and paralytic ileus (OR = 16; p = 0.033) were independent findings suggesting life-threatening mesenteric occlusion. The ancillary CT findings occur with different frequencies in SMAT and SMVT. However, the independent findings indicating life-threatening mesenteric occlusion are diminished bowel wall enhancement and paralytic ileus.

  14. The hybrid four-CBS-domain KINβγ subunit functions as the canonical γ subunit of the plant energy sensor SnRK1.

    Science.gov (United States)

    Ramon, Matthew; Ruelens, Philip; Li, Yi; Sheen, Jen; Geuten, Koen; Rolland, Filip

    2013-07-01

    The AMPK/SNF1/SnRK1 protein kinases are a family of ancient and highly conserved eukaryotic energy sensors that function as heterotrimeric complexes. These typically comprise catalytic α subunits and regulatory β and γ subunits, the latter function as the energy-sensing modules of animal AMPK through adenosine nucleotide binding. The ability to monitor accurately and adapt to changing environmental conditions and energy supply is essential for optimal plant growth and survival, but mechanistic insight in the plant SnRK1 function is still limited. In addition to a family of γ-like proteins, plants also encode a hybrid βγ protein that combines the Four-Cystathionine β-synthase (CBS)-domain (FCD) structure in γ subunits with a glycogen-binding domain (GBD), typically found in β subunits. We used integrated functional analyses by ectopic SnRK1 complex reconstitution, yeast mutant complementation, in-depth phylogenetic reconstruction, and a seedling starvation assay to show that only the hybrid KINβγ protein that recruited the GBD around the emergence of the green chloroplast-containing plants, acts as the canonical γ subunit required for heterotrimeric complex formation. Mutagenesis and truncation analysis further show that complex interaction in plant cells and γ subunit function in yeast depend on both a highly conserved FCD and a pre-CBS domain, but not the GBD. In addition to novel insight into canonical AMPK/SNF/SnRK1 γ subunit function, regulation and evolution, we provide a new classification of plant FCD genes as a convenient and reliable tool to predict regulatory partners for the SnRK1 energy sensor and novel FCD gene functions. © 2013 The Authors The Plant Journal © 2013 John Wiley & Sons Ltd.

  15. The Regulation of NF-κB Subunits by Phosphorylation

    Directory of Open Access Journals (Sweden)

    Frank Christian

    2016-03-01

    Full Text Available The NF-κB transcription factor is the master regulator of the inflammatory response and is essential for the homeostasis of the immune system. NF-κB regulates the transcription of genes that control inflammation, immune cell development, cell cycle, proliferation, and cell death. The fundamental role that NF-κB plays in key physiological processes makes it an important factor in determining health and disease. The importance of NF-κB in tissue homeostasis and immunity has frustrated therapeutic approaches aimed at inhibiting NF-κB activation. However, significant research efforts have revealed the crucial contribution of NF-κB phosphorylation to controlling NF-κB directed transactivation. Importantly, NF-κB phosphorylation controls transcription in a gene-specific manner, offering new opportunities to selectively target NF-κB for therapeutic benefit. This review will focus on the phosphorylation of the NF-κB subunits and the impact on NF-κB function.

  16. DNA binding properties of the small cascade subunit Csa5.

    Directory of Open Access Journals (Sweden)

    Michael Daume

    Full Text Available CRISPR-Cas systems provide immunity against viral attacks in archaeal and bacterial cells. Type I systems employ a Cas protein complex termed Cascade, which utilizes small CRISPR RNAs to detect and degrade the exogenic DNA. A small sequence motif, the PAM, marks the foreign substrates. Previously, a recombinant type I-A Cascade complex from the archaeon Thermoproteus tenax was shown to target and degrade DNA in vitro, dependent on a native PAM sequence. Here, we present the biochemical analysis of the small subunit, Csa5, of this Cascade complex. T. tenax Csa5 preferentially bound ssDNA and mutants that showed decreased ssDNA-binding and reduced Cascade-mediated DNA cleavage were identified. Csa5 oligomerization prevented DNA binding. Specific recognition of the PAM sequence was not observed. Phylogenetic analyses identified Csa5 as a universal member of type I-A systems and revealed three distinct groups. A potential role of Csa5 in R-loop stabilization is discussed.

  17. NMDA receptor structures reveal subunit arrangement and pore architecture.

    Science.gov (United States)

    Lee, Chia-Hsueh; Lü, Wei; Michel, Jennifer Carlisle; Goehring, April; Du, Juan; Song, Xianqiang; Gouaux, Eric

    2014-07-10

    N-methyl-d-aspartate (NMDA) receptors are Hebbian-like coincidence detectors, requiring binding of glycine and glutamate in combination with the relief of voltage-dependent magnesium block to open an ion conductive pore across the membrane bilayer. Despite the importance of the NMDA receptor in the development and function of the brain, a molecular structure of an intact receptor has remained elusive. Here we present X-ray crystal structures of the Xenopus laevis GluN1-GluN2B NMDA receptor with the allosteric inhibitor, Ro25-6981, partial agonists and the ion channel blocker, MK-801. Receptor subunits are arranged in a 1-2-1-2 fashion, demonstrating extensive interactions between the amino-terminal and ligand-binding domains. The transmembrane domains harbour a closed-blocked ion channel, a pyramidal central vestibule lined by residues implicated in binding ion channel blockers and magnesium, and a ∼twofold symmetric arrangement of ion channel pore loops. These structures provide new insights into the architecture, allosteric coupling and ion channel function of NMDA receptors.

  18. NMDA receptor structures reveal subunit arrangement and pore architecture

    Science.gov (United States)

    Lee, Chia-Hsueh; Lü, Wei; Michel, Jennifer Carlisle; Goehring, April; Du, Juan; Song, Xianqiang; Gouaux, Eric

    2014-01-01

    Summary N-methyl-d-aspartate (NMDA) receptors are Hebbian-like coincidence detectors, requiring binding of glycine and glutamate in combination with the relief of voltage-dependent magnesium block to open an ion conductive pore across the membrane bilayer. Despite the importance of the NMDA receptor in the development and function of the brain, a molecular structure of an intact receptor has remained elusive. Here we present x-ray crystal structures of the GluN1/GluN2B NMDA receptor with the allosteric inhibitor, Ro25-6981, partial agonists and the ion channel blocker, MK-801. Receptor subunits are arranged in a 1-2-1-2 fashion, demonstrating extensive interactions between the amino terminal and ligand binding domains. The transmembrane domains harbor a closed-blocked ion channel, a pyramidal central vestibule lined by residues implicated in binding ion channel blockers and magnesium, and a ~2-fold symmetric arrangement of ion channel pore loops. These structures provide new insights into the architecture, allosteric coupling and ion channel function of NMDA receptors. PMID:25008524

  19. Binding of cholera toxin B subunit to intestinal epithelial cells.

    Science.gov (United States)

    Navolotskaya, Elena V; Sadovnikov, Vladimir B; Lipkin, Valery M; Zav'yalov, Vladimir P

    2018-03-01

    We have prepared 125 I-labeled cholera toxin B subunit ( 125 I-labeled CT-B, a specific activity of 98Ci/mmol) and found that it binds to rat IEC-6 and human Caco-2 intestinal epithelial cells with high affinity (K d 3.6 and 3.7nM, respectively). The binding of labeled protein was completely inhibited by unlabeled thymosin-α 1 (TM-α 1 ), interferon-α 2 (IFN-α 2 ), and the synthetic peptide LKEKK that corresponds to residues 16-20 in TM-α 1 and 131-135 in IFN-α 2 , but was not inhibited by the synthetic peptide KKEKL with inverted amino acid sequence (K i >10μM). Thus, TM-α 1 , IFN-α 2 , and the peptide: LKEKK bind with high affinity and specificity to the cholera toxin receptor on IEC-6 and Caco-2 cells. It was found that CT-B and the peptide: LKEKK at concentrations of 10-1000nM increased in a dose-dependent manner the nitric oxide production and the soluble guanylate cyclase activity in IEC-6 and Caco-2 cells. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Vaccine profile of herpes zoster (HZ/su) subunit vaccine.

    Science.gov (United States)

    Cunningham, Anthony L; Heineman, Thomas

    2017-07-01

    Herpes zoster (HZ) causes an often severe and painful rash in older people and may be complicated by prolonged pain (postherpetic neuralgia; PHN) and by dissemination in immune-compromised patients. HZ results from reactivation of latent varicella-zoster virus (VZV) infection, often associated with age-related or other causes of decreased T cell immunity. A live attenuated vaccine boosts this immunity and provides partial protection against HZ, but this decreases with age and declines over 8 years. Areas covered: A new HZ subunit (HZ/su) vaccine combines a key surface VZV glycoprotein (E) with a T cell-boosting adjuvant system (AS01 B ) and is administered by two intramuscular injections two months apart. Expert commentary: HZ/su showed excellent efficacy of ~90% in immunocompetent adults ≥50 and ≥70 years of age, respectively, in the ZOE-50 and ZOE-70 phase III controlled trials. Efficacy was unaffected by advancing age and persisted for >3 years. Approximately 9.5% of subjects had severe, but transient (1-2 days) injection site pain, swelling or redness. Compliance with both vaccine doses was high (95%). The vaccine will have a major impact on HZ management. Phase I-II trials showed safety and immunogenicity in severely immunocompromised patients. Phase III trial results are expected soon.

  1. Design of a hyperstable 60-subunit protein icosahedron

    Science.gov (United States)

    Hsia, Yang; Bale, Jacob B.; Gonen, Shane; Shi, Dan; Sheffler, William; Fong, Kimberly K.; Nattermann, Una; Xu, Chunfu; Huang, Po-Ssu; Ravichandran, Rashmi; Yi, Sue; Davis, Trisha N.; Gonen, Tamir; King, Neil P.; Baker, David

    2016-07-01

    The icosahedron is the largest of the Platonic solids, and icosahedral protein structures are widely used in biological systems for packaging and transport. There has been considerable interest in repurposing such structures for applications ranging from targeted delivery to multivalent immunogen presentation. The ability to design proteins that self-assemble into precisely specified, highly ordered icosahedral structures would open the door to a new generation of protein containers with properties custom-tailored to specific applications. Here we describe the computational design of a 25-nanometre icosahedral nanocage that self-assembles from trimeric protein building blocks. The designed protein was produced in Escherichia coli, and found by electron microscopy to assemble into a homogenous population of icosahedral particles nearly identical to the design model. The particles are stable in 6.7 molar guanidine hydrochloride at up to 80 degrees Celsius, and undergo extremely abrupt, but reversible, disassembly between 2 molar and 2.25 molar guanidinium thiocyanate. The icosahedron is robust to genetic fusions: one or two copies of green fluorescent protein (GFP) can be fused to each of the 60 subunits to create highly fluorescent ‘standard candles’ for use in light microscopy, and a designed protein pentamer can be placed in the centre of each of the 20 pentameric faces to modulate the size of the entrance/exit channels of the cage. Such robust and customizable nanocages should have considerable utility in targeted drug delivery, vaccine design and synthetic biology.

  2. Fc receptor gamma subunit polymorphisms and systemic lupus erythematosus

    International Nuclear Information System (INIS)

    Al-Ansari, Aliya; Ollier, W.E.; Gonzalez-Gay, Miguel A.; Gul, Ahmet; Inanac, Murat; Ordi, Jose; Teh, Lee-Suan; Hajeer, Ali H.

    2004-01-01

    To investigate the possible association between Fc receptor gamma polymorphisms and systemic lupus erythematosus (SLE). We have investigated the full FcR gamma gene for polymorphisms using polymerase chain reaction (PCR)-single strand confirmational polymorphisms and DNA sequencing .The polymorphisms identified were genotype using PCR-restriction fragment length polymorphism. Systemic lupus erythematosus cases and controls were available from 3 ethnic groups: Turkish, Spanish and Caucasian. The study was conducted in the year 2001 at the Arthritis Research Campaign, Epidemiology Unit, Manchester University Medical School, Manchester, United Kingdom. Five single nucleotide polymorphisms were identified, 2 in the promoter, one in intron 4 and, 2 in the 3'UTR. Four of the 5 single nucleotide polymorphisms (SNPs) were relatively common and investigated in the 3 populations. Allele and genotype frequencies of all 4 investigated SNPs were not statistically different cases and controls. fc receptor gamma gene does not appear to contribute to SLE susceptibility. The identified polymorphisms may be useful in investigating other diseases where receptors containing the FcR gamma subunit contribute to the pathology. (author)

  3. Self-Subunit Swapping Occurs in Another Gene Type of Cobalt Nitrile Hydratase

    Science.gov (United States)

    Xia, Yuanyuan; Cui, Youtian; Kobayashi, Michihiko; Zhou, Zhemin

    2012-01-01

    Self-subunit swapping is one of the post-translational maturation of the cobalt-containing nitrile hydratase (Co-NHase) family of enzymes. All of these NHases possess a gene organization of , which allows the activator protein to easily form a mediatory complex with the α-subunit of the NHase after translation. Here, we discovered that the incorporation of cobalt into another type of Co-NHase, with a gene organization of , was also dependent on self-subunit swapping. We successfully isolated a recombinant NHase activator protein (P14K) of Pseudomonas putida NRRL-18668 by adding a Strep-tag N-terminal to the P14K gene. P14K was found to form a complex [α(StrepP14K)2] with the α-subunit of the NHase. The incorporation of cobalt into the NHase of P. putida was confirmed to be dependent on the α-subunit substitution between the cobalt-containing α(StrepP14K)2 and the cobalt-free NHase. Cobalt was inserted into cobalt-free α(StrepP14K)2 but not into cobalt-free NHase, suggesting that P14K functions not only as a self-subunit swapping chaperone but also as a metallochaperone. In addition, NHase from P. putida was also expressed by a mutant gene that was designed with a order. Our findings expand the general features of self-subunit swapping maturation. PMID:23226397

  4. Separation of ribosomal subunits of Escherichia coli by Sepharose chromatography using reverse salt gradient.

    Science.gov (United States)

    Kirillov, S V; Makhno, V I; Peshin, N N; Semenkov, Y P

    1978-11-01

    A mixture of 30 S and 50 S subunits quantitatively absorbs on a column of Sepharose--4B from the buffer: 0.02 M Tris--HCl, pH 7.5, containing 1.5 M (NH4)2SO4. During elution by reverse gradient of ammonium sulphate (1.5--0.05 M) the subunits are eluted at different salt concentrations. Complete separation of subunits is attained in the absence of Mg2+ ions. The 30 S subunits prepared from 70 S ribosomes according to this procedure are fully active in the codon--dependent binding of a specific aminoacyl--tRNA. After their reassociation with 50 S subunits isolated by zonal centrifugation, the resulting 70 S ribosomes are active in polypeptide synthesis at the same degree as control 70 S ribosomes in which both types of subunits were prepared by zonal centrifugation. The initial 70 S ribosomes for the chromatographic separation into subunits can be obtained by their pelleting from a crude extract with subsequent washing with concentrated solutions of NH4Cl in the ultracentrifuge, or by salt fractionation of the crude extract according to a slightly modified procedure of Kurland.

  5. Effect of chronic ethanol consumption on the subunit structure of the mitochondrial ATP synthase

    Energy Technology Data Exchange (ETDEWEB)

    Coleman, W.B.; Spach, P.I.; Cunningham, C.C. (Wake Forest Univ., Winston-Salem, NC (United States))

    1991-03-11

    The relative concentrations of several subunits of the mitochondrial F{sub 0}F{sub 1} ATP synthase were determined in hepatic mitochondria and submitochrondrial particles (SMP) isolated from ethanol-fed and control rats. Animals were maintained on an ethanol-containing liquid diet for 31 days. The polypeptides were analyzed by densitometry measurements of SDS-polyacrylamide gel electrophoresis patterns. Subunit 8 was decreased 31% in intact mitochondria, whereas subunit 6 could not be measured. In contrast, there were no significant ethanol-related depressions in subunits {alpha}, {beta} and OSCP of the F{sub 0}F{sub 1} or the adenine nucleotide carrier (AdNC) in intact mitochondria. In ethanol SMP subunits 6 and 8 were decreased 41 and 34%, respectively; subunits {alpha}, {beta}, {gamma} and OSCP were also present in lowered amounts due to the loose attachment of F{sub 1} to F{sub 0}. The remainder of the F{sub 1} was observed in the soluble fraction resulting from preparation of SMP. The AdNC was present in normal amounts in ethanol SMP. These results demonstrate that ethanol consumption causes a decrease in the content of mitochondrial synthesized subunits 6 and 8 whereas no effect is exerted on the concentrations of nuclear gene products of the ATP synthase complex. Likewise, the adenine nucleotide transporter, also a nuclear gene product, is unaffected by ethanol consumption.

  6. Subunit conformational variation within individual GroEL oligomers resolved by Cryo-EM.

    Science.gov (United States)

    Roh, Soung-Hun; Hryc, Corey F; Jeong, Hyun-Hwan; Fei, Xue; Jakana, Joanita; Lorimer, George H; Chiu, Wah

    2017-08-01

    Single-particle electron cryo-microscopy (cryo-EM) is an emerging tool for resolving structures of conformationally heterogeneous particles; however, each structure is derived from an average of many particles with presumed identical conformations. We used a 3.5-Å cryo-EM reconstruction with imposed D7 symmetry to further analyze structural heterogeneity among chemically identical subunits in each GroEL oligomer. Focused classification of the 14 subunits in each oligomer revealed three dominant classes of subunit conformations. Each class resembled a distinct GroEL crystal structure in the Protein Data Bank. The conformational differences stem from the orientations of the apical domain. We mapped each conformation class to its subunit locations within each GroEL oligomer in our dataset. The spatial distributions of each conformation class differed among oligomers, and most oligomers contained 10-12 subunits of the three dominant conformation classes. Adjacent subunits were found to more likely assume the same conformation class, suggesting correlation among subunits in the oligomer. This study demonstrates the utility of cryo-EM in revealing structure dynamics within a single protein oligomer.

  7. Voltage-Gated Sodium Channel β1/β1B Subunits Regulate Cardiac Physiology and Pathophysiology

    Directory of Open Access Journals (Sweden)

    Nnamdi Edokobi

    2018-04-01

    Full Text Available Cardiac myocyte contraction is initiated by a set of intricately orchestrated electrical impulses, collectively known as action potentials (APs. Voltage-gated sodium channels (NaVs are responsible for the upstroke and propagation of APs in excitable cells, including cardiomyocytes. NaVs consist of a single, pore-forming α subunit and two different β subunits. The β subunits are multifunctional cell adhesion molecules and channel modulators that have cell type and subcellular domain specific functional effects. Variants in SCN1B, the gene encoding the Nav-β1 and -β1B subunits, are linked to atrial and ventricular arrhythmias, e.g., Brugada syndrome, as well as to the early infantile epileptic encephalopathy Dravet syndrome, all of which put patients at risk for sudden death. Evidence over the past two decades has demonstrated that Nav-β1/β1B subunits play critical roles in cardiac myocyte physiology, in which they regulate tetrodotoxin-resistant and -sensitive sodium currents, potassium currents, and calcium handling, and that Nav-β1/β1B subunit dysfunction generates substrates for arrhythmias. This review will highlight the role of Nav-β1/β1B subunits in cardiac physiology and pathophysiology.

  8. Topographic Studies of Torpedo Acetylcholine Receptor Subunits as a Transmembrane Complex

    Science.gov (United States)

    Strader, Catherine D.; Raftery, Michael A.

    1980-10-01

    The exposure of the four subunits of the acetylcholine receptor from Torpedo californica on both the extracellular and cytoplasmic faces of the postsynaptic membranes of the electroplaque cells has been investigated. Sealed membrane vesicles containing no protein components other than the receptor were isolated and were shown to have 95% of their synaptic surfaces facing the medium. The susceptibility of the four receptor subunits in these preparations to hydrolysis by trypsin both from the external and from the internal medium was used to investigate the exposure of the subunits on the synaptic and cytoplasmic surfaces of the membrane. It was shown by sodium dodecyl sulfate gel electrophoresis of the tryptic products that all four subunits are exposed on the extracellular surface to a similar degree. All four subunits are also exposed on the internal surface of the membrane, but the apparent degree of exposure varies with the subunit size, the larger subunits being more exposed. The results are discussed in terms of a possible topographic model of the receptor as a transmembrane protein complex.

  9. Expression of gill vacuolar-type H+-ATPase B subunit, and Na+, K+-ATPase alpha- and beta- subunit messenger RNAs in smolting Salmo salar

    DEFF Research Database (Denmark)

    Seidelin, Michel; Madsen, Steffen; Cutler, Christopher P

    2001-01-01

    Changes in gill vacuolar-type H+-ATPase B subunit, and Na+,K+-ATPase alpha and beta subunit mRNA expression were examined during the course of smoltification in Salmo salar. We cloned and sequenced cDNA fragments of S. salar gill i) vacuolar-type H+-ATPase (V-H+-ATPase) B subunit, ii) Na+,K+-ATPase....... The peak smelt stage was, however, characterized by simultaneously elevated gill Na+,K+-ATPase expression and low V-H+-ATPase expression, and possibly ensures the complete transformation of the gill into a hypo-osmoregulatory organ and hence the development of optimal SW-tolerance of the smolt....... alpha (1) subunit, and iii) Na+,K+-ATPase beta (1) subunit, and used these as Northern blotting probes. During smoltification, the salmon showed a typical increase in gill Na+,K+-ATPase activity and improved hypo-osmoregulatory ability as judged by their ability to regulate plasma [Cl-] in a 24-hr...

  10. An alternating GluN1-2-1-2 subunit arrangement in mature NMDA receptors.

    Directory of Open Access Journals (Sweden)

    Morgane Riou

    Full Text Available NMDA receptors (NMDARs form glutamate-gated ion channels that play a critical role in CNS physiology and pathology. Together with AMPA and kainate receptors, NMDARs are known to operate as tetrameric complexes with four membrane-embedded subunits associating to form a single central ion-conducting pore. While AMPA and some kainate receptors can function as homomers, NMDARs are obligatory heteromers composed of homologous but distinct subunits, most usually of the GluN1 and GluN2 types. A fundamental structural feature of NMDARs, that of the subunit arrangement around the ion pore, is still controversial. Thus, in a typical NMDAR associating two GluN1 and two GluN2 subunits, there is evidence for both alternating 1/2/1/2 and non-alternating 1/1/2/2 arrangements. Here, using a combination of electrophysiological and cross-linking experiments, we provide evidence that functional GluN1/GluN2A receptors adopt the 1/2/1/2 arrangement in which like subunits are diagonal to one another. Moreover, based on the recent crystal structure of an AMPA receptor, we show that in the agonist-binding and pore regions, the GluN1 subunits occupy a "proximal" position, closer to the central axis of the channel pore than that of GluN2 subunits. Finally, results obtained with reducing agents that differ in their membrane permeability indicate that immature (intracellular and functional (plasma-membrane inserted pools of NMDARs can adopt different subunit arrangements, thus stressing the importance of discriminating between the two receptor pools in assembly studies. Elucidating the quaternary arrangement of NMDARs helps to define the interface between the subunits and to understand the mechanism and pharmacology of these key signaling receptors.

  11. Regulated appearance of NMDA receptor subunits and channel functions during in vitro neuronal differentiation.

    Science.gov (United States)

    Jelitai, Márta; Schlett, Katalin; Varju, Patrícia; Eisel, Ulrich; Madarász, Emília

    2002-04-01

    The schedule of NMDA receptor subunit expression and the appearance of functional NMDA-gated ion channels were investigated during the retinoic acid (RA) induced neuronal differentiation of NE-4C, a p53-deficient mouse neuroectodermal progenitor cell line. NR2A, NR2B, and NR2D subunit transcripts were present in both nondifferentiated and neuronally differentiated cultures, while NR2C subunits were expressed only transiently, during the early period of neural differentiation. Several splice variants of NR1 were detected in noninduced progenitors and in RA-induced cells, except the N1 exon containing transcripts that appeared after the fourth day of induction, when neuronal processes were already formed. NR1 and NR2A subunit proteins were detected both in nondifferentiated progenitor cells and in neurons, while the mature form of NR2B subunit protein appeared only at the time of neuronal process elongation. Despite the early presence of NR1 and NR2A subunits, NMDA-evoked responses could be detected in NE-4C neurons only after the sixth day of induction, coinciding in time with the expression of the mature NR2B subunit. The formation of functional NMDA receptors also coincided with the appearance of synapsin I and synaptophysin. The lag period between the production of the subunits and the onset of channel function suggests that subunits capable of channel formation cannot form functional NMDA receptors until a certain stage of neuronal commitment. Thus, the in vitro neurogenesis by NE-4C cells provides a suitable tool to investigate some inherent regulatory processes involved in the initial maturation of NMDA receptor complexes. Copyright 2002 Wiley Periodicals, Inc.

  12. Three human alcohol dehydrogenase subunits: cDNA structure and molecular and evolutionary divergence

    International Nuclear Information System (INIS)

    Ikuta, T.; Szeto, S.; Yoshida, A.

    1986-01-01

    Class I human alcohol dehydrogenase (ADH; alcohol:NAD + oxidoreductase, EC 1.1.1.1) consists of several homo- and heterodimers of α, β, and γ subunits that are governed by the ADH1, ADH2, and ADH3 loci. The authors previously cloned a full length of cDNA for the β subunit, and the complete sequence of 374 amino acid residues was established. cDNAs for the α and γ subunits were cloned and characterized. A human liver cDNA library, constructed in phage λgt11, was screened by using a synthetic oligonucleotide probe that was matched to the γ but not to the β sequence. Clone pUCADHγ21 and clone pUCADHα15L differed from β cDNA with respect to restriction sites and hybridization with the nucleotide probe. Clone pUCADHγ21 contained an insertion of 1.5 kilobase pairs (kbp) and encodes 374 amino acid residues compatible with the reported amino acid sequence of the γ subunit. Clone pUCADHα15L contained an insertion of 2.4 kbp and included nucleotide sequences that encode 374 amino acid residues for another subunit, the γ subunit. In addition, this clone contained the sequences that encode the COOH-terminal part of the β subunit at its extended 5' region. The amino acid sequences and coding regions of the cDNAs of the three subunits are very similar. A high degree of resemblance is observed also in their 3' noncoding regions. However, distinctive differences exist in the vicinity of the Zn-binding cysteine residue at position 46. Based on the cDNA sequences and the deduced amino acid sequences of the three subunits, their structural and evolutionary relationships are discussed

  13. Expression of BK Ca channels and the modulatory beta-subunits in the rat and porcine trigeminal ganglion

    DEFF Research Database (Denmark)

    Johansson, Helle Wulf; Hay-Schmidt, Anders; Poulsen, Asser Nyander

    2009-01-01

    (Ca) channel protein was visualized by western blotting and histochemistry. The presence of the modulatory beta1-beta 4 subunit mRNAs was investigated using RT-PCR. beta1-, beta2- and beta 4-subunit mRNAs were expressed in rat TG whereas beta2- and beta 4-subunits were detected in porcine TG. Western blotting...

  14. Expression of Gill Vacuolar-Type H^+-ATPase B Subunit, and Na^+, K^+-ATPase α_1 and β_1 Subunit Messenger RNAs in Smolting Salmo salar(Physiology)

    OpenAIRE

    Michel, Seidelin; Steffen S., Madsen; Christopher P., Cutler; Gordon, Cramb; Institute of Biology, University of Southern Denmark-Main Campus : Odense University:(Present address)Department of Life Sciences and Chemistry, Roskilde University; Institute of Biology, University of Southern Denmark-Main Campus : Odense University; School of Biomedical Sciences, Bute Medical Buildings, University of St. Andrews; School of Biomedical Sciences, Bute Medical Buildings, University of St. Andrews

    2001-01-01

    Changes in gill vacuolar-type HT-ATPase B subunit, and Na^+,K^+-ATPase α and β subunit mRNA expression were examined during the course of smoltification in Salmo salar. We cloned and sequenced cDNA fragments of S. salar gill i) vacuolar-type H^+-ATPase (V-H^+-ATPase) B subunit, ii) Na^+,K^+-ATPase α_1 subunit, and iii) Na^+,K^+-ATPase β_1 subunit, and used these as Northern blotting probes. During smoltification, the salmon showed a typical increase in gill Na^+,K^+-ATPase activity and improv...

  15. Modulation of nucleotide binding to the catalytic sites of thermophilic F(1)-ATPase by the epsilon subunit: implication for the role of the epsilon subunit in ATP synthesis.

    Science.gov (United States)

    Yasuno, Taichi; Muneyuki, Eiro; Yoshida, Masasuke; Kato-Yamada, Yasuyuki

    2009-12-11

    Effect of epsilon subunit on the nucleotide binding to the catalytic sites of F(1)-ATPase from the thermophilic Bacillus PS3 (TF(1)) has been tested by using alpha(3)beta(3)gamma and alpha(3)beta(3)gammaepsilon complexes of TF(1) containing betaTyr341 to Trp substitution. The nucleotide binding was assessed with fluorescence quenching of the introduced Trp. The presence of the epsilon subunit weakened ADP binding to each catalytic site, especially to the highest affinity site. This effect was also observed when GDP or IDP was used. The ratio of the affinity of the lowest to the highest nucleotide binding sites had changed two orders of magnitude by the epsilon subunit. The differences may relate to the energy required for the binding change in the ATP synthesis reaction and contribute to the efficient ATP synthesis.

  16. Molecular cloning of the α subunit of human and guinea pig leukocyte adhesion glycoprotein Mo1: Chromosomal localization and homology to the α subunits of integrins

    International Nuclear Information System (INIS)

    Arnaout, M.A.; Remold-O'Donnell, E.; Pierce, M.W.; Harris, P.; Tenen, D.G.

    1988-01-01

    The cell surface-glycoprotein Mo1 is a member of the family of leukocyte cell adhesion molecules (Leu-CAMs) that includes lymphocyte function-associated antigen 1 (LFA-1) and p150,95. Each Leu-CAM is a heterodimer with a distinct α subunit noncovalently associated with a common β subunit. The authors describe the isolation and analysis of two partial cDNA clones encoding the α subunit of the Leu-CAM Mo1 in humans and guinea pigs. A monoclonal antibody directed against an epitope in the carboxyl-terminal portion of the guinea pig α chain was used for immunoscreening a λgt11 expression library. The sequence of a 378-base-pair insert from one immunoreactive clone revealed a single continuous open reading frame encoding 126 amino acids including a 26-amino acid tryptic peptide isolated from the purified guinea pig α subunit. A cDNA clone of identical size was isolated from a human monocyte/lymphocyte cDNA library by using the guinea pig clone as a probe. The human clone also encoded a 126-amino acid peptide including the sequence of an additional tryptic peptide present in purified human Mo1α chain. Southern analysis of DNA from hamster-human hybrids localized the human Mo1α chain to chromosome 16, which has been shown to contain the gene for the α chain of lymphocyte function-associated antigen 1. These data suggest that the α subunits of Leu-CAMs evolved by gene duplication from a common ancestral gene and strengthen the hypothesis that the α subunits of these heterodimeric cell adhesion molecules on myeloid and lymphoid cells, platelets, and fibroblasts are evolutionary related

  17. Use of an α3-β4 nicotinic acetylcholine receptor subunit concatamer to characterize ganglionic receptor subtypes with specific subunit composition reveals species-specific pharmacologic properties

    Science.gov (United States)

    Stokes, Clare; Papke, Roger L.

    2012-01-01

    Drug development for nicotinic acetylcholine receptors (nAChR) is challenged by subtype diversity arising from variations in subunit composition. On-target activity for neuronal heteromeric receptors is typically associated with CNS receptors that contain α4 and other subunits, while off-target activity could be associated with ganglionic-type receptors containing α3β4 binding sites and other subunits, including β4, β2, α5, or α3 as a structural subunit in the pentamer. Additional interest in α3 β4 α5-containing receptors arises from genome-wide association studies linking these genes, and a single nucleotide polymorphism (SNP) in α5 in particular, to lung cancer and heavy smoking. While α3 and β4 readily form receptors in expression system such as the Xenopus oocyte, since α5 is not required for function, simple co-expression approaches may under-represent α5-containing receptors. We used a concatamer of human α3 and β4 subunits to form ligand-binding domains, and show that we can force the insertions of alternative structural subunits into the functional pentamers. These α3β4 variants differ in sensitivity to ACh, nicotine, varenicline, and cytisine. Our data indicated lower efficacy for varenicline and cytisine than expected for β4-containing receptors, based on previous studies of rodent receptors. We confirm that these therapeutically important α4 receptor partial agonists may present different autonomic-based side-effect profiles in humans than will be seen in rodent models, with varenicline being more potent for human than rat receptors and cytisine less potent. Our initial characterizations failed to find functional effects of the α5 SNP. However, our data validate this approach for further investigations. PMID:22580377

  18. Genetic Analysis of the Mode of Interplay between an ATPase Subunit and Membrane Subunits of the Lipoprotein-Releasing ATP-Binding Cassette Transporter LolCDE†

    OpenAIRE

    Ito, Yasuko; Matsuzawa, Hitomi; Matsuyama, Shin-ichi; Narita, Shin-ichiro; Tokuda, Hajime

    2006-01-01

    The LolCDE complex, an ATP-binding cassette (ABC) transporter, releases lipoproteins from the inner membrane, thereby initiating lipoprotein sorting to the outer membrane of Escherichia coli. The LolCDE complex is composed of two copies of an ATPase subunit, LolD, and one copy each of integral membrane subunits LolC and LolE. LolD hydrolyzes ATP on the cytoplasmic side of the inner membrane, while LolC and/or LolE recognize and release lipoproteins anchored to the periplasmic leaflet of the i...

  19. Characterization of the alpha and beta subunits of casein kinase 2 by far-UV CD spectroscopy

    DEFF Research Database (Denmark)

    Issinger, O G; Brockel, C; Boldyreff, B

    1992-01-01

    , comparison of the far-UV CD spectrum of reconstituted CK-2 with the spectra of the subunits indicates that conformational changes occur in the backbone region upon association. Such changes may explain the increased enzyme activity of the holoenzyme relative to that of the alpha subunit itself. In contrast......, no changes in the far-UV CD spectrum of the alpha subunit are observed in the presence of casein or the synthetic decapeptide substrate RRRDDDSDDD. Furthermore, the alpha-helical structure of the alpha subunit (but not the beta subunit) can be increased in the presence of stoichiometric amounts of heparin...

  20. Distinct Subunit Domains Govern Synaptic Stability and Specificity of the Kainate Receptor

    Directory of Open Access Journals (Sweden)

    Christoph Straub

    2016-07-01

    Full Text Available Synaptic communication between neurons requires the precise localization of neurotransmitter receptors to the correct synapse type. Kainate-type glutamate receptors restrict synaptic localization that is determined by the afferent presynaptic connection. The mechanisms that govern this input-specific synaptic localization remain unclear. Here, we examine how subunit composition and specific subunit domains contribute to synaptic localization of kainate receptors. The cytoplasmic domain of the GluK2 low-affinity subunit stabilizes kainate receptors at synapses. In contrast, the extracellular domain of the GluK4/5 high-affinity subunit synergistically controls the synaptic specificity of kainate receptors through interaction with C1q-like proteins. Thus, the input-specific synaptic localization of the native kainate receptor complex involves two mechanisms that underlie specificity and stabilization of the receptor at synapses.

  1. Alpha subunit and human chorionic gonadotropin in normal pregnancy and gestational trophoblastic disease.

    Science.gov (United States)

    Elegbe, R A; Pattillo, R A; Hussa, R O; Hoffmann, R G; Damole, I O; Finlayson, W E

    1984-03-01

    The reported incidence of gestational trophoblastic disease is an order of magnitude higher in Nigeria than in the United States. Sera from a total of 283 pregnant black patients, 138 United States and 148 Nigerian pregnant patients, were analyzed for their serum levels of alpha subunit and human chorionic gonadotropin (hCG). The patterns of hCG secretion were similar in the two populations during normal pregnancy. However, the level of alpha subunit was persistently higher in Nigerian women than in comparable pregnant United States patients. A statistically significantly higher alpha subunit level in the Nigerian patients was found only in the ten- to 13-week gestational period (P less than .005). The higher level of alpha subunit in pregnancy in Nigerian women may signal a population of trophoblastic cells which may be at higher risk for malignancy development in the Nigerian woman.

  2. Subunit interactions in tyrosinase from frog epidermis in immobilized enzyme systems.

    Science.gov (United States)

    Iborra, J L; Ferragut, J A; Lozano, J A

    1981-01-01

    1. Frog epidermis tyrosinase was coupled to Sepharose activated with low concentrations of CNBr. The tetrameric form of the enzyme was linked to the matrix via its subunits. Dissociation of the bound active enzyme with guanidinium chloride yielded an active immobilized dimeric derivative. 2. Immobilized dimeric derivative was able to interact with soluble subunits formed transiently during renaturation. An 85% recovery of the native dopa oxidase specific activity was achieved after hybridization. 3. Fluorescence spectra of different immobilized derivatives suggested that tryptophan residues were involved in the interactions between tyrosinase subunits. 4. It is suggested that the activation of the subunits of tyrosinase involves a conformational change towards a more unfolded state, which favours a reassociation to the dimeric active state. Images Fig. 2. PMID:6798971

  3. Sulfation of the human chorionic gonadotropin α subunit in placental explants

    International Nuclear Information System (INIS)

    Bielinska, M.; Birken, S.; Boime, I.

    1986-01-01

    The asparagine-linked oligosaccharides of bovine lutropin, terminate with sulfate while the oligosaccharides of human chorionic gonadotropin contain terminal sialic acid. Using a cell-free system to study sulfation of gonadotropins, the authors unexpectedly observed that asialo hCG or its subunits could serve as substrates for in vitro sulfation. To investigate as possible physiological role of sulfate in the expression of hCG, first trimester placental explants were incubated with [/sup 35/S]O/sub 4/ in Krebs-Ringer-bicarbonate solution. The media were immunoprecipitated with antisera against α or β subunits of hCG and the proteins were subjected to SDS-polyacrylamide gel electrophoresis. Both free and dimer α subunits contained sulfate. No detectable sulfation of β subunit occurred under our labelling

  4. 78 FR 8102 - Kootenai National Forest; Buckhorn Planning Subunit; Lincoln County, Montana; Environmental...

    Science.gov (United States)

    2013-02-05

    ... Forest Service Kootenai National Forest; Buckhorn Planning Subunit; Lincoln County, Montana; Environmental Impact Statement AGENCY: Forest Service, USDA. ACTION: Notice of intent to prepare an environmental impact statement. SUMMARY: The Forest Service will prepare an Environmental Impact Statement (EIS...

  5. Multiple roles of Rev3, the catalytic subunit of polzeta in maintaining genome stability in vertebrates

    NARCIS (Netherlands)

    E. Sonoda (Eiichiro); S. Takeda (Shiunichi); T. Okada (Takashi); G.Y. Zhao (Guang); S. Tateishi (Satoshi); K. Araki (Kasumi); M. Yamaizumi (Masaru); T. Yagi (Takashi); N.S. Verkaik (Nicole); D.C. van Gent (Dik); M. Takata (Minoru)

    2003-01-01

    textabstractTranslesion DNA synthesis (TLS) and homologous DNA recombination (HR) are two major postreplicational repair (PRR) pathways. The REV3 gene of Saccharomyces cerevisiae encodes the catalytic subunit of DNA polymerase zeta, which is involved in mutagenic TLS. To

  6. Pharmacological consequences of the coexpression of BK channel α and auxiliary β subunits

    Science.gov (United States)

    Torres, Yolima P.; Granados, Sara T.; Latorre, Ramón

    2014-01-01

    Coded by a single gene (Slo1, KCM) and activated by depolarizing potentials and by a rise in intracellular Ca2+ concentration, the large conductance voltage- and Ca2+-activated K+ channel (BK) is unique among the superfamily of K+ channels. BK channels are tetramers characterized by a pore-forming α subunit containing seven transmembrane segments (instead of the six found in voltage-dependent K+ channels) and a large C terminus composed of two regulators of K+ conductance domains (RCK domains), where the Ca2+-binding sites reside. BK channels can be associated with accessory β subunits and, although different BK modulatory mechanisms have been described, greater interest has recently been placed on the role that the β subunits may play in the modulation of BK channel gating due to its physiological importance. Four β subunits have currently been identified (i.e., β1, β2, β3, and β4) and despite the fact that they all share the same topology, it has been shown that every β subunit has a specific tissue distribution and that they modify channel kinetics as well as their pharmacological properties and the apparent Ca2+ sensitivity of the α subunit in different ways. Additionally, different studies have shown that natural, endogenous, and synthetic compounds can modulate BK channels through β subunits. Considering the importance of these channels in different pathological conditions, such as hypertension and neurological disorders, this review focuses on the mechanisms by which these compounds modulate the biophysical properties of BK channels through the regulation of β subunits, as well as their potential therapeutic uses for diseases such as those mentioned above. PMID:25346693

  7. Crystal Structure of the Oxazolidinone Antibiotic Linezolid Bound to the 50S Ribosomal Subunit

    Energy Technology Data Exchange (ETDEWEB)

    Ippolito,J.; Kanyo, Z.; Wang, D.; Franceschi, F.; Moore, P.; Steitz, T.; Duffy, E.

    2008-01-01

    The oxazolidinone antibacterials target the 50S subunit of prokaryotic ribosomes. To gain insight into their mechanism of action, the crystal structure of the canonical oxazolidinone, linezolid, has been determined bound to the Haloarcula marismortui 50S subunit. Linezolid binds the 50S A-site, near the catalytic center, which suggests that inhibition involves competition with incoming A-site substrates. These results provide a structural basis for the discovery of improved oxazolidinones active against emerging drug-resistant clinical strains.

  8. Neutron Scattering and the 30 S Ribosomal Subunit of E. Coli

    Science.gov (United States)

    Moore, P. B.; Engelman, D. M.; Langer, J. A.; Ramakrishnan, V. R.; Schindler, D. G.; Schoenborn, B. P.; Sillers, I. Y.; Yabuki, S.

    1982-06-01

    This paper reviews the progress made in the study of the internal organization of the 30 S ribosomal subunit of E. coli by neutron scattering since 1975. A map of that particle showing the position of 14 of the subunit's 21 proteins is presented, and the methods currently used for collecting and analyzing such data are discussed. Also discussed is the possibility of extending the interpretation of neutron mapping data beyond the limits practical today.

  9. Small Subunits of Serine Palmitoyltransferase (ssSPTs) and Their Physiological Roles

    Science.gov (United States)

    2014-02-12

    found predominantly in fungal and plant species and the other in mammals, insects and nematodes. Interestingly a sphingolipid biosynthetic pathway...subunits belong to two distinct clades. Within each clade, the mammalian, metazoan, nematode, fungal and plant subunits show tight clustering. Probably...30688- 94 7. Berkey R, Bendigeri D, Xiao S. 2012. Sphingolipids and plant defense/ disease : the "death" connection and beyond. Frontiers in plant

  10. Over-expressions of AMPK subunits in ovarian carcinomas with significant clinical implications

    International Nuclear Information System (INIS)

    Li, Cuilan; Liu, Vincent WS; Chiu, Pui M; Chan, David W; Ngan, Hextan YS

    2012-01-01

    AMP-activated protein kinase (AMPK) has recently been considered as a potential target for cancer therapy. However, the expression status of various subunits of the heterotrimeric AMPK in human cancers is rarely reported. We decided to determine their expressions in ovarian carcinomas and their relationships with the disease. Expressions and locations of the AMPK-α1, -α2, -β1, -β2, -γ1 and -γ2 were detected by quantitative PCR (Q-PCR) and immunohistochemical staining (IHC). Their expression levels in ovarian tumors were compared with normal controls and also correlated with clinicopathological parameters. Except AMPK-α1, expressions of the other five AMPK subunits are significantly higher in ovarian carcinomas as determined by Q-PCR. Although IHC detection of AMPK-γ1 and -γ2 were not successful, over-expressions of AMPK-α2, -β1, and -β2 were further confirmed by IHC. Over-expressions of various AMPK subunits occurred independently and were mainly detected in the cytoplasm. Interestingly, AMPK-α2 and -β1 were also detected in the nucleus and cell membrane, respectively. Clinical correlation analyses indicate that expressions of different AMPK subunits are associated with different subtypes of carcinoma. High expression of AMPK-α2 is significantly associated with endometrioid carcinomas. On the other hand, high expressions of AMPK-β and -γ subunits are associated with mucinous and serous carcinomas, respectively. Furthermore, high expressions of AMPK-β1 and -γ2 are also associated with early and late stages of disease, respectively. Finally, patients with high expression of AMPK-α2 had better prognosis. Aberrant expressions of AMPK subunits may play important roles in ovarian carcinogenesis. Each AMPK subunit may have its own function other than just a component of the AMPK molecule. Correlations with clinical parameters suggest that expressions of AMPK subunits have different clinical implications in ovarian cancer development

  11. Nicotinic acetylcholine receptor: subunit structure, functional binding sites, and ion transport properties

    International Nuclear Information System (INIS)

    Raftery, M.A.; Dunn, S.M.J.; Conti-Tronconi, B.M.; Middlemas, D.S.; Crawford, R.D.

    1983-01-01

    The structure of the nicotinic acetylcholine receptor has been highly conserved during animal evolution, and in all the species and tissues studied so far, including mammals, it is a pseudosymmetric, pentameric complex of related subunits with very similar physical properties. All subunits of these nicotinic receptors were derived from a common ancestral gene, probably by way of gene duplications occurring very early in animal evolution. 45 refs., 8 figs., 2 tabs

  12. The subunit structure of methylmalonyl-CoA mutase from Propionibacterium shermanii.

    OpenAIRE

    Francalanci, F; Davis, N K; Fuller, J Q; Murfitt, D; Leadlay, P F

    1986-01-01

    5'-Deoxyadenosylcobalamin-dependent methylmalonyl-CoA mutase was purified to homogeneity from Propionibacterium shermanii by a simplified procedure. The native enzyme has an apparent Mr of 165,000, similar to the enzyme from other sources but larger than previously reported. It consists of two non-identical subunits, of Mr 79,000 and 67,000 respectively. The smaller subunit is apparently not a proteolytic fragment of the larger one. The final preparation usually contained some inactive mutase...

  13. More β subunits of F1 catalyze during oxidative phosphorylation than during ATP hydrolysis

    International Nuclear Information System (INIS)

    Wang, J.H.; Wu, J.C.; Cesana, J.

    1987-01-01

    Bovine heart mitochondrial F 1 -ATPase was labeled specifically with 7-chloro-4-nitro-2,3,1-[ 14 C]benzodiazole (NBD-F 1 on Tyr-311 in either the β' subunit or the β'' subunits to form O-β'-NBD-F 1 or O-β''-NBD-F 1 . The labeled F 1 was used to combine with F 1 -deficient submitochondrial particles (ASU) to form O-β'-NBD-F 1 -ASU and O-β''-NBD-F 1 -ASU respectively. It was found that O-β'-NBD-F 1 -ASU retains a higher percentage of catalytic activity for oxidative phosphorylation than for ATP hydrolysis; whereas O-β''-NED-F 1 -ASU retains a higher percentage of catalytic activity for ATP hydrolysis than for oxidative phosphorylation. Preincubation of O-β'-NBD-F 1 -ASu in the assay medium for oxidative phosphorylation did not significantly increase its catalytic activity for ATP hydrolysis. These observations show that only the β' subunit catalyzes ATP hydrolysis directly, whereas all three β-subunits catalyze oxidative phosphorylation and that during oxidative phosphorylation the β' and β'' subunits do not switch places with each other. A possible explanation of those results is that interactions between the subunits could provide only the site on β' subunit with the unique facility for rapid ATP hydrolysis, whereas during oxidative phosphorylation the reverse reaction could be effected by protein conformation changes driven by proton flux in all three β subunits so that phosphorylation could occur at all three sites

  14. Structural domains of the human GABAA receptor β3 subunit involved in the actions of pentobarbital

    Science.gov (United States)

    Serafini, Ruggero; Bracamontes, John; Steinbach, Joe Henry

    2000-01-01

    This study was conducted to search for the residues of the β3 subunit which affect pentobarbital action on the γ-aminobutyric acid type A (GABAA) receptor. Three chimeras were constructed by joining the GABAA receptor β3 subunit to the ρ1 subunit. For each chimera, the N-terminal sequence was derived from the β3 subunit and the C-terminal sequence from the ρ1 subunit, with junctions located between the membrane-spanning regions M2 and M3, in the middle of M2, or in M1, respectively.In receptors obtained by the coexpression of α1 with the chimeric subunits, in contrast with those obtained by the coexpression of α1 and β3, pentobarbital exhibited lower potentiation of GABA-evoked responses, and in the direct gating of Cl− currents, an increase in the EC50 together with a marked decrease in the relative maximal efficacy compared with that of GABA.Estimates of the channel opening probability through variance analysis and single-channel recordings of one chimeric subunit showed that the reduced relative efficacy for gating largely resulted from an increase in gating by GABA, with little change in efficacy of pentobarbital.A fit of the time course of the response by the predictions of a class of reaction schemes is consistent with the conclusion that the change in the concentration dependence of activation by pentobarbital is due to a change in pentobarbital affinity for the receptor. Therefore, the data suggest that residues of the β3 subunit involved in pentobarbital binding to GABAA receptors are located downstream from the middle of the M2 region. PMID:10790149

  15. CHRISGAS Project. WP13: Ancillary and Novel Processes. Final Report: Separation of Hydrogen with Membranes Combined with Water Gas Shift Reaction

    Energy Technology Data Exchange (ETDEWEB)

    Sanchez-Hervas, J. M.; Marono, M.; Barreiro, M. M.

    2011-05-13

    Oxygen pressurized gasification of biomass out stands as a very promising approach to obtain energy or hydrogen from renewable sources. The technical feasibility of this technology has been investigated under the scope of the VI FP CHRISGAS project, which started in September 2004 and had a duration of five and a half years. The Division of Combustion and Gasification of CIEMAT participated in this project in Work Package 13: Ancillary and novel processes, studying innovative gas separation and gas upgrading systems. Such systems include novel or available high temperature water gas shift catalysts and commercially available membranes not yet tried in this type of atmosphere. This report describes the activities carried out during the project regarding the performance of high temperature water gas shift catalysts for upgrading of synthesis gas obtained from biomass gasification, the separation of H2 with selective membranes and the combination of both processes in one by means of a catalytic membrane reactor. (Author) 20 refs.

  16. CHRISGAS Project. WP13: Ancillary and Novel Processes. Final Report: Separation of Hydrogen with Membranes Combined with Water Gas Shift Reaction

    International Nuclear Information System (INIS)

    Sanchez-Hervas, J. M.; Marono, M.; Barreiro, M. M.

    2011-01-01

    Oxygen pressurized gasification of biomass out stands as a very promising approach to obtain energy or hydrogen from renewable sources. The technical feasibility of this technology has been investigated under the scope of the VI FP CHRISGAS project, which started in September 2004 and had a duration of five and a half years. The Division of Combustion and Gasification of CIEMAT participated in this project in Work Package 13: Ancillary and novel processes, studying innovative gas separation and gas upgrading systems. Such systems include novel or available high temperature water gas shift catalysts and commercially available membranes not yet tried in this type of atmosphere. This report describes the activities carried out during the project regarding the performance of high temperature water gas shift catalysts for upgrading of synthesis gas obtained from biomass gasification, the separation of H2 with selective membranes and the combination of both processes in one by means of a catalytic membrane reactor. (Author) 20 refs.

  17. Cell block samples from endobronchial ultrasound transbronchial needle aspiration provide sufficient material for ancillary testing in lung cancer-a quaternary referral centre experience.

    Science.gov (United States)

    Hopkins, Emily; Moffat, David; Parkinson, Ian; Robinson, Peter; Jersmann, Hubertus; Dougherty, Brendan; Birader, Mohammed I; Francis, Kate; Nguyen, Phan

    2016-09-01

    Rapid on site examination (ROSE) is encouraged at endobronchial ultrasound transbronchial needles aspiration (EBUS-TBNA) to improve diagnostic yield. Due to new therapeutic options in lung cancer, it is not sufficient to merely distinguish between non-small cell lung carcinoma (NSCLC) and small cell lung carcinoma (SCLC). Immunohistochemistry (IHC) distinction is now standard practice, as well as additional molecular testing where clinically indicated. We investigated the diagnostic yield of on-site smears vs . cell block and the provision of cellular material for ancillary testing at our centre. A retrospective audit of all EBUS-TBNA procedures performed until July 2012 was undertaken. Diagnostic yield on smears versus cell block was recorded. Cell blocks were reviewed by an experienced pathologist to determine diagnostic accuracy and whether IHC and molecular testing were possible. In total, 234 procedures were recorded with 101 (43.2%) malignant cases, 107 (45.7%) benign cases and an initial 26/234 (11.1%) insufficient for diagnosis of which 11/234 (4.7%) were false negatives for malignancy after further follow up. The average number of passes was 4.5. For malignancies, smear diagnosis was possible in 95% (96/101) of cases and cell block diagnosis in 93.5% (87/93) of cases. There was sufficient material for IHC in 97.7% (85/87) of malignant cases. In 79.3% (69/87) of NSCLCs molecular testing for epidermal growth factor receptor (EGFR) mutation analysis was theoretically possible on samples obtained. Cell blocks are not inferior to smears for diagnostic accuracy and provide sufficient samples for histology. However, ROSE assists the physician on how best to manage samples for ancillary testing.

  18. Fluorescence In Situ Hybridization for MDM2 Amplification as a Routine Ancillary Diagnostic Tool for Suspected Well-Differentiated and Dedifferentiated Liposarcomas: Experience at a Tertiary Center

    Directory of Open Access Journals (Sweden)

    Khin Thway

    2015-01-01

    Full Text Available Background. The assessment of MDM2 gene amplification by fluorescence in situ hybridization (FISH has become a routine ancillary tool for diagnosing atypical lipomatous tumor (ALT/well-differentiated liposarcoma and dedifferentiated liposarcoma (WDL/DDL in specialist sarcoma units. We describe our experience of its utility at our tertiary institute. Methods. All routine histology samples in which MDM2 amplification was assessed with FISH over a 2-year period were included, and FISH results were correlated with clinical and histologic findings. Results. 365 samples from 347 patients had FISH for MDM2 gene amplification. 170 were positive (i.e., showed MDM2 gene amplification, 192 were negative, and 3 were technically unsatisfactory. There were 122 histologically benign cases showing a histology:FISH concordance rate of 92.6%, 142 WDL/DDL (concordance 96.5%, and 34 cases histologically equivocal for WDL (concordance 50%. Of 64 spindle cell/pleomorphic neoplasms (in which DDL was a differential diagnosis, 21.9% showed MDM2 amplification. Of the cases with discrepant histology and FISH, all but 3 had diagnoses amended following FISH results. For discrepancies of benign histology but positive FISH, lesions were on average larger, more frequently in “classical” (intra-abdominal or inguinal sites for WDL/DDL and more frequently core biopsies. Discrepancies of malignant histology but negative FISH were smaller, less frequently in “classical” sites but again more frequently core biopsies. Conclusions. FISH has a high correlation rate with histology for cases with firm histologic diagnoses of lipoma or WDL/DDL. It is a useful ancillary diagnostic tool in histologically equivocal cases, particularly in WDL lacking significant histologic atypia or DDL without corresponding WDL component, especially in larger tumors, those from intra-abdominal or inguinal sites or core biopsies. There is a significant group of well-differentiated adipocytic neoplasms

  19. Theoretical Insight into the Spectral Characteristics of Fe(II-Based Complexes for Dye-Sensitized Solar Cells—Part I: Polypyridyl Ancillary Ligands

    Directory of Open Access Journals (Sweden)

    Xiaoqing Lu

    2011-01-01

    Full Text Available The design of light-absorbent dyes with cheaper, safer, and more sustainable materials is one of the key issues for the future development of dye-sensitized solar cells (DSSCs. We report herein a theoretical investigation on a series of polypyridyl Fe(II-based complexes of FeL2(SCN2, [FeL3]2+, [FeL′(SCN3]-, [FeL′2]2+, and FeL′′(SCN2 (L = 2,2′-bipyridyl-4,4′-dicarboxylic acid, L′ = 2,2′,2″-terpyridyl-4,4′,4″-tricarboxylic acid, L″ = 4,4‴-dimethyl-2,2′ : 6′,2″ :6″,2‴-quaterpyridyl-4′,4″-biscarboxylic acid by density functional theory (DFT and time-dependent DFT (TD-DFT. Molecular geometries, electronic structures, and optical absorption spectra are predicted in both the gas phase and methyl cyanide (MeCN solution. Our results show that polypyridyl Fe(II-based complexes display multitransition characters of Fe → polypyridine metal-to-ligand charge transfer and ligand-to-ligand charge transfer in the range of 350–800 nm. Structural optimizations by choosing different polypyridyl ancillary ligands lead to alterations of the molecular orbital energies, oscillator strength, and spectral response range. Compared with Ru(II sensitizers, Fe(II-based complexes show similar characteristics and improving trend of optical absorption spectra along with the introduction of different polypyridyl ancillary ligands.

  20. Theoretical study on the influence of ancillary ligand on the energy and optical properties of heteroleptic phosphorescent Ir(III) complexes

    Energy Technology Data Exchange (ETDEWEB)

    Yang Baozhu [Faculty of Chemistry and Chemical Engineering, Changzhou University, Changzhou 213164 (China); State Key Laboratory of Theoretical and Computational Chemistry, Institute of Theoretical Chemistry, Jilin University, Changchun 130023 (China); Zhang Min [Changchun Institute of Applied Chemistry, Changchun 130022 (China); State Key Laboratory of Theoretical and Computational Chemistry, Institute of Theoretical Chemistry, Jilin University, Changchun 130023 (China); Zhang Hongxing, E-mail: zhanghx@mail.jlu.edu.c [State Key Laboratory of Theoretical and Computational Chemistry, Institute of Theoretical Chemistry, Jilin University, Changchun 130023 (China); Sun Jiazhong [State Key Laboratory of Theoretical and Computational Chemistry, Institute of Theoretical Chemistry, Jilin University, Changchun 130023 (China)

    2011-06-15

    The geometries, energies, and electronic properties of a series of phosphorescent Ir(III) complexes including FIrpic, FIrmpic, FIrpca, and FIrprza have been characterized within density functional theory DFT calculations, which can reproduce and rationalize experimental results. The properties of excited states of the Ir(III) complexes were characterized by the configuration interaction with single-excitation (CIS) method. The ground- and excited-state geometries were optimized at the B3LYP/LANL2DZ and CIS/LANL2DZ levels, respectively. The absorption and phosphorescence wavelengths were computed based on the optimized ground- and excited-state geometries, respectively, by the time-dependent density functional theory (TD-DFT) methods. All the energies have been calculated by B3LYP method. The predictions revealed that the nature of the ancillary ligands can influence the distributions of frontier molecular orbitals and their energies, resulting in impact on the transition character and change in the emission color. In addition, the charge of transport quality has been estimated approximately by the predicted reorganization energy ({lambda}). Our result also indicates that the substituent groups and different auxiliary ligand not only change the character of transition but also affect the rate and balance of charge transfer. - Highlights: {yields} The geometries, energies, and electronic properties of a series of phosphorescent Ir(III) complexes including FIrpic, FIrmpic, FIrpca, and FIrprza have been characterized with DFT calculations. {yields} The predictions revealed that the nature of the ancillary ligands can influence the distributions of frontier molecular orbitals and their energies, resulting in impact on the transition character and change in the emission color. {yields} Our result also indicates that the substitute groups and different auxiliary ligand not only change the character of transition but also affect the rate and balance of charge transfer.

  1. Modulatory mechanisms and multiple functions of somatodendritic A-type K+ channel auxiliary subunits

    Directory of Open Access Journals (Sweden)

    Henry Hungtao Jerng

    2014-03-01

    Full Text Available Auxiliary subunits are non-conducting, modulatory components of the multi-protein ion channel complexes that underlie normal neuronal signaling. They interact with the pore-forming α-subunits to modulate surface distribution, ion conductance, and channel gating properties. For the somatodendritic subthreshold A-type potassium (ISA channel based on Kv4 α-subunits, two types of auxiliary subunits have been extensively studied: Kv channel-interacting proteins (KChIPs and dipeptidyl peptidase-like proteins (DPLPs. KChIPs are cytoplasmic calcium-binding proteins that interact with intracellular portions of the Kv4 subunits, whereas DPLPs are type II transmembrane proteins that associate with the Kv4 channel core. Both KChIPs and DPLPs genes contain multiple start sites that are used by various neuronal populations to drive the differential expression of functionally distinct N-terminal variants. In turn, these N-terminal variants generate tremendous functional diversity across the nervous system. Here, we focus our review on (1 the molecular mechanism underlying the unique properties of different N-terminal variants, (2 the shaping of native ISA properties by the concerted actions of KChIPs and DPLP variants, and (3 the surprising ways that KChIPs and DPLPs coordinate the activity of multiple channels to fine-tune neuronal excitability. Unlocking the unique contributions of different auxiliary subunit N-terminal variants may provide an important opportunity to develop novel targeted therapeutics to treat numerous neurological disorders.

  2. Functional regulation of BK potassium channels by γ1 auxiliary subunits.

    Science.gov (United States)

    Gonzalez-Perez, Vivian; Xia, Xiao-Ming; Lingle, Christopher J

    2014-04-01

    Many K(+) channels are oligomeric complexes with intrinsic structural symmetry arising from the homo-tetrameric core of their pore-forming subunits. Allosteric regulation of tetramerically symmetric proteins, whether by intrinsic sensing domains or associated auxiliary subunits, often mirrors the fourfold structural symmetry. Here, through patch-clamp recordings of channel population ensembles and also single channels, we examine regulation of the Ca(2+)- and voltage-activated large conductance Ca(2+)-activated K(+) (BK) channel by associated γ1-subunits. Through expression of differing ratios of γ1:α-subunits, the results reveal an all-or-none functional regulation of BK channels by γ-subunits: channels either exhibit a full gating shift or no shift at all. Furthermore, the γ1-induced shift exhibits a state-dependent labile behavior that recapitulates the fully shifted or unshifted behavior. The γ1-induced shift contrasts markedly to the incremental shifts in BK gating produced by 1-4 β-subunits and adds a new layer of complexity to the mechanisms by which BK channel functional diversity is generated.

  3. Regulatory role of voltage-gated sodium channel β subunits in sensory neurons

    Directory of Open Access Journals (Sweden)

    Mohamed eChahine

    2011-11-01

    Full Text Available Voltage-gated Na+ channels are transmembrane-bound proteins incorporating aqueous conduction pores that are highly selective for Na+. The opening of these channels results in the rapid influx of Na+ ions that depolarize the cell and drive the rapid upstroke of nerve and muscle action potentials. While the concept of a Na+-selective ion channel had been formulated in the 1940s, it was not until the 1980s that the biochemical properties of the 260-kDa and 36-kDa auxiliary β subunits (β1, β2 were first described. Subsequent cloning and heterologous expression studies revealed that the  subunit forms the core of the channel and is responsible for both voltage-dependent gating and ionic selectivity. To date, ten isoforms of the Na+ channel α subunit have been identified that vary in their primary structures, tissue distribution, biophysical properties, and sensitivity to neurotoxins. Four β subunits (β1-β4 and two splice variants (β1A, β1B have been identified that modulate the subcellular distribution, cell surface expression, and functional properties of the α subunits. The purpose of this review is to provide a broad overview of β subunit expression and function in peripheral sensory neurons and examine their contributions to neuropathic pain.

  4. Mechanisms of the scaffold subunit in facilitating protein phosphatase 2A methylation.

    Directory of Open Access Journals (Sweden)

    Vitali Stanevich

    Full Text Available The function of the biologically essential protein phosphatase 2A (PP2A relies on formation of diverse heterotrimeric holoenzymes, which involves stable association between PP2A scaffold (A and catalytic (C or PP2Ac subunits and binding of variable regulatory subunits. Holoenzyme assembly is highly regulated by carboxyl methylation of PP2Ac-tail; methylation of PP2Ac and association of the A and C subunits are coupled to activation of PP2Ac. Here we showed that PP2A-specific methyltransferase, LCMT-1, exhibits a higher activity toward the core enzyme (A-C heterodimer than free PP2Ac, and the A-subunit facilitates PP2A methylation via three distinct mechanisms: 1 stabilization of a proper protein fold and an active conformation of PP2Ac; 2 limiting the space of PP2Ac-tail movement for enhanced entry into the LCMT-1 active site; and 3 weak electrostatic interactions between LCMT-1 and the N-terminal HEAT repeats of the A-subunit. Our results revealed a new function and novel mechanisms of the A-subunit in PP2A methylation, and coherent control of PP2A activity, methylation, and holoenzyme assembly.

  5. A molecular breadboard: Removal and replacement of subunits in a hepatitis B virus capsid.

    Science.gov (United States)

    Lee, Lye Siang; Brunk, Nicholas; Haywood, Daniel G; Keifer, David; Pierson, Elizabeth; Kondylis, Panagiotis; Wang, Joseph Che-Yen; Jacobson, Stephen C; Jarrold, Martin F; Zlotnick, Adam

    2017-11-01

    Hepatitis B virus (HBV) core protein is a model system for studying assembly and disassembly of icosahedral structures. Controlling disassembly will allow re-engineering the 120 subunit HBV capsid, making it a molecular breadboard. We examined removal of subunits from partially crosslinked capsids to form stable incomplete particles. To characterize incomplete capsids, we used two single molecule techniques, resistive-pulse sensing and charge detection mass spectrometry. We expected to find a binomial distribution of capsid fragments. Instead, we found a preponderance of 3 MDa complexes (90 subunits) and no fragments smaller than 3 MDa. We also found 90-mers in the disassembly of uncrosslinked HBV capsids. 90-mers seem to be a common pause point in disassembly reactions. Partly explaining this result, graph theory simulations have showed a threshold for capsid stability between 80 and 90 subunits. To test a molecular breadboard concept, we showed that missing subunits could be refilled resulting in chimeric, 120 subunit particles. This result may be a means of assembling unique capsids with functional decorations. © 2017 The Protein Society.

  6. The 2.3 {angstrom} crystal structure of cholera toxin B subunit pentamer: Choleragenoid

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Rong-Guang; Westbrook, M.L. [Argonne National Lab., IL (United States); Maulik, P.R.; Reed, R.A.; Shipley, G. [Boston Univ., MA (United States). School of Medicine; Westbrook, E.M. [Argonne National Lab., IL (United States)]|[Northwestern Univ., Evanston, IL (United States); Scott, D.L.; Otwinowski, Z. [Yale Univ., New Haven, CT (United States)

    1996-02-01

    Cholera toxin, a heterohexameric AB{sub 5} enterotoxin released by Vibrio cholera, induces a profuse secretory diarrhea in susceptible hosts. Choleragenoid, the B subunit pentamer of cholera toxin, directs the enzymatic A subunit to its target by binding to GM{sub 1} gangliosides exposed on the luminal surface of intestinal epithelial cells. We have solved the crystal structure of choleragenoid at 2.3 {Angstrom} resolution by combining single isomorphous replacement with non-crystallographic symmetry averaging. The structure of the B subunits, and their pentameric arrangement, closely resembles that reported for the intact holotoxin (choleragen), the heat-labile enterotoxin from E. coli, and for a choleragenoid-GM{sub 1} pentasaccharide complex. In the absence of the A subunit the central cavity of the B pentamer is a highly solvated channel. The binding of the A subunit or the receptor pentasaccharide to choleragenoid has only a modest effect on the local stereochemistry and does not perceptibly alter the subunit interface.

  7. LACK, a RACK1 ortholog, facilitates cytochrome c oxidase subunit expression to promote Leishmania major fitness.

    Science.gov (United States)

    Cardenas, Daviel; Carter, Pamela M; Nation, Catherine S; Pizarro, Juan C; Guidry, Jessie; Aiyar, Ashok; Kelly, Ben L

    2015-04-01

    Leishmania are kinetoplastid parasites that cause the sandfly-transmitted disease leishmaniasis. To maintain fitness throughout their infectious life cycle, Leishmania must undergo rapid metabolic adaptations to the dramatically distinct environments encountered during transition between sandfly and vertebrate hosts. We performed proteomic and immunoblot analyses of attenuated L. major strains deficient for LACK, the Leishmania ortholog of the mammalian receptor for activated c kinase (RACK1), that is important for parasite thermotolerance and virulence. This approach identified cytochrome c oxidase (LmCOX) subunit IV as a LACK-dependent fitness protein. Consistent with decreased levels of LmCOX subunit IV at mammalian temperature, and in amastigotes, LmCOX activity and mitochondrial function were also impaired in LACK-deficient L. major under these conditions. Importantly, overexpression of LmCOX subunit IV in LACK-deficient L. major restored thermotolerance and macrophage infectivity. Interestingly, overexpression of LmCOX subunit IV enhanced LmCOX subunit VI expression at mammalian temperature. Collectively, our data suggest LACK promotes Leishmania adaptation to the mammalian host environment by sustaining LmCOX subunit IV expression and hence energy metabolism in response to stress stimuli such as heat. These findings extend the repertoire of RACK1 protein utility to include a role in mitochondrial function. © 2015 John Wiley & Sons Ltd.

  8. High resolution solution structure of the 1.3S subunit of transcarboxylase from Propionibacterium shermanii.

    Science.gov (United States)

    Reddy, D V; Shenoy, B C; Carey, P R; Sönnichsen, F D

    2000-03-14

    Transcarboxylase (TC) from Propionibacterium shermanii, a biotin-dependent enzyme, catalyzes the transfer of a carboxyl group from methylmalonyl-CoA to pyruvate to form propionyl-CoA and oxalacetate. Within the multi-subunit enzyme complex, the 1.3S subunit functions as the carboxyl group carrier and also binds the other two subunits to assist in the overall assembly of the enzyme. The 1.3S subunit is a 123 amino acid polypeptide (12.6 kDa) to which biotin is covalently attached at Lys 89. The three-dimensional solution structure of the full-length holo-1.3S subunit of TC has been solved by multidimensional heteronuclear NMR spectroscopy. The C-terminal half of the protein (51-123) is folded into a compact all-beta-domain comprising of two four-stranded antiparallel beta-sheets connected by short loops and turns. The fold exhibits a high 2-fold internal symmetry and is similar to that of the biotin carboxyl carrier protein (BCCP) of acetyl-CoA carboxylase, but lacks an extension that has been termed "protruding thumb" in BCCP. The first 50 residues, which have been shown to be involved in intersubunit interactions in the intact enzyme, appear to be disordered in the isolated 1.3S subunit. The molecular surface of the folded domain has two distinct surfaces: one side is highly charged, while the other comprises mainly hydrophobic, highly conserved residues.

  9. The elusive third subunit IIa of the bacterial B-type oxidases: the enzyme from the hyperthermophile Aquifex aeolicus.

    Directory of Open Access Journals (Sweden)

    Laurence Prunetti

    Full Text Available The reduction of molecular oxygen to water is catalyzed by complicated membrane-bound metallo-enzymes containing variable numbers of subunits, called cytochrome c oxidases or quinol oxidases. We previously described the cytochrome c oxidase II from the hyperthermophilic bacterium Aquifex aeolicus as a ba(3-type two-subunit (subunits I and II enzyme and showed that it is included in a supercomplex involved in the sulfide-oxygen respiration pathway. It belongs to the B-family of the heme-copper oxidases, enzymes that are far less studied than the ones from family A. Here, we describe the presence in this enzyme of an additional transmembrane helix "subunit IIa", which is composed of 41 amino acid residues with a measured molecular mass of 5105 Da. Moreover, we show that subunit II, as expected, is in fact longer than the originally annotated protein (from the genome and contains a transmembrane domain. Using Aquifex aeolicus genomic sequence analyses, N-terminal sequencing, peptide mass fingerprinting and mass spectrometry analysis on entire subunits, we conclude that the B-type enzyme from this bacterium is a three-subunit complex. It is composed of subunit I (encoded by coxA(2 of 59000 Da, subunit II (encoded by coxB(2 of 16700 Da and subunit IIa which contain 12, 1 and 1 transmembrane helices respectively. A structural model indicates that the structural organization of the complex strongly resembles that of the ba(3 cytochrome c oxidase from the bacterium Thermus thermophilus, the IIa helical subunit being structurally the lacking N-terminal transmembrane helix of subunit II present in the A-type oxidases. Analysis of the genomic context of genes encoding oxidases indicates that this third subunit is present in many of the bacterial oxidases from B-family, enzymes that have been described as two-subunit complexes.

  10. Preclinical and clinical development of a dengue recombinant subunit vaccine.

    Science.gov (United States)

    Manoff, Susan B; George, Sarah L; Bett, Andrew J; Yelmene, Michele L; Dhanasekaran, Govindarajan; Eggemeyer, Linda; Sausser, Michele L; Dubey, Sheri A; Casimiro, Danilo R; Clements, David E; Martyak, Timothy; Pai, Vidya; Parks, D Elliot; Coller, Beth-Ann G

    2015-12-10

    This review focuses on a dengue virus (DENV) vaccine candidate based on a recombinant subunit approach which targets the DENV envelope glycoprotein (E). Truncated versions of E consisting of the N-terminal portion of E (DEN-80E) have been expressed recombinantly in the Drosophila S2 expression system and shown to have native-like conformation. Preclinical studies demonstrate that formulations containing tetravalent DEN-80E adjuvanted with ISCOMATRIX™ adjuvant induce high titer virus neutralizing antibodies and IFN-γ producing T cells in flavivirus-naïve non-human primates. The preclinical data further suggest that administration of such formulations on a 0, 1, 6 month schedule may result in higher maximum virus neutralizing antibody titers and better durability of those titers compared to administration on a 0, 1, 2 month schedule. In addition, the virus neutralizing antibody titers induced by adjuvanted tetravalent DEN-80E compare favorably to the titers induced by a tetravalent live virus comparator. Furthermore, DEN-80E was demonstrated to be able to boost virus neutralizing antibody titers in macaques that have had a prior DENV exposure. A monovalent version of the vaccine candidate, DEN1-80E, was formulated with Alhydrogel™ and studied in a proof-of-principle Phase I clinical trial by Hawaii Biotech, Inc. (NCT00936429). The clinical trial results demonstrate that both the 10 μg and 50 μg formulations of DEN1-80E with 1.25 mg of elemental aluminum were immunogenic when administered in a 3-injection series (0, 1, 2 months) to healthy, flavivirus-naïve adults. The vaccine formulations induced DENV-1 neutralizing antibodies in the majority of subjects, although the titers in most subjects were modest and waned over time. Both the 10 μg DEN1-80E and the 50 μg DEN1-80E formulations with Alhydrogel™ were generally well tolerated. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Subunits VIIa,b,c of human cytochrome c oxidase. Identification of both 'heart-type' and 'liver-type' isoforms of subunit VIIa in human heart

    NARCIS (Netherlands)

    van Kuilenburg, A. B.; van Beeumen, J. J.; van der Meer, N. M.; Muijsers, A. O.

    1992-01-01

    The N-terminal amino acid sequences and the electrophoretic mobilities of the subunits VIIa, VIIb and VIIc of cytochrome c oxidase purified from human heart were investigated and compared with those from human skeletal muscle and from bovine heart. In purified human heart cytochrome c oxidase, both

  12. The α' subunit of β-conglycinin and various glycinin subunits of soy are not required to modulate hepatic lipid metabolism in rats.

    Science.gov (United States)

    Chatterjee, Cynthia; Liu, Jiajie; Wood, Carla; Gagnon, Christine; Cober, Elroy R; Frégeau-Reid, Judith A; Gleddie, Stephen; Xiao, Chao-Wu

    2017-03-21

    This study examined the effect of soy proteins with depletion of different subunits of the two major storage proteins, β-conglycinin and glycinin, on hepatic lipids and proteins involved in lipid metabolism in rats, since the bioactive component of soy responsible for lipid-lowering is unclear. Weanling Sprague Dawley rats were fed diets containing either 20% casein protein in the absence (casein) or presence (casein + ISF) of isoflavones or 20% alcohol-washed soy protein isolate (SPI) or 20% soy protein concentrates derived from a conventional (Haro) or 2 soybean lines lacking the α' subunit of β-conglycinin and the A1-3 (1TF) or A1-5 (1a) subunits of glycinin. After 8 weeks, the rats were necropsied and liver proteins and lipids were extracted and analysed. The results showed that soy protein diets reduced lipid droplet accumulation and content in the liver compared to casein diets. The soy protein diets also decreased the level of hepatic mature SREBP-1 and FAS in males, with significant decreases in diets 1TF and 1a compared to the casein diets. The effect of the soy protein diets on female hepatic mature SREBP-1, FAS, and HMGCR was confounded since casein + ISF decreased these levels compared to casein alone perhaps muting the decrease by soy protein. A reduction in both phosphorylated and total STAT3 in female livers by ISF may account for the gender difference in mechanism in the regulation and protein expression of the lipid modulators. Overall, soy protein deficient in the α' subunit of β-conglycinin and A1-5 subunits of glycinin maintain similar hypolipidemic function compared to the conventional soy protein. The exact bioactive component(s) warrant identification.

  13. Yeast inner-subunit PA-NZ-1 labeling strategy for accurate subunit identification in a macromolecular complex through cryo-EM analysis.

    Science.gov (United States)

    Wang, Huping; Han, Wenyu; Takagi, Junichi; Cong, Yao

    2018-04-03

    Cryo-electron microscopy (cryo-EM) has been established as one of the central tools in the structural study of macromolecular complexes. Although intermediate- or low-resolution structural information through negative staining (NS) or cryo-EM analysis remains highly valuable, we lack general and efficient ways to achieve unambiguous subunit identification in these applications. Here, we took advantage of the extremely high affinity between a dodecapeptide "PA" tag and the NZ-1 antibody Fab fragment to develop an efficient "yeast inner-subunit PA-NZ-1 labeling" (YISPANL) strategy that when combined with cryo-EM could precisely identify subunits in macromolecular complexes. Using this strategy combined with cryo-EM 3D reconstruction, we were able to visualize the characteristic NZ-1 Fab density attached to the PA tag inserted into a surface-exposed loop in the middle of the sequence of CCT6 subunit present in the S. cerevisiae group II chaperonin TRiC/CCT. This procedure facilitated the unambiguous localization of CCT6 in the TRiC complex. The PA tag was designed to contain only 12 amino acids and a tight turn configuration; when inserted into a loop, it usually has a high chance of maintaining the epitope structure and low likelihood of perturbing the native structure and function of the target protein compared to other tagging systems. We also found the association between PA and NZ-1 can sustain the cryo freezing conditions, resulting in very high occupancy of the Fab in the final cryo-EM images. Our study demonstrated the robustness of this strategy combined with cryo-EM in efficient and accurate subunit identification in challenging multi-component complexes. Copyright © 2018. Published by Elsevier Ltd.

  14. Dithiothreitol activation of the insulin receptor/kinase does not involve subunit dissociation of the native α2β2 insulin receptor subunit complex

    International Nuclear Information System (INIS)

    Sweet, L.J.; Wilden, P.A.; Pessin, J.E.

    1986-01-01

    The subunit composition of the dithiothreitol- (DTT) activated insulin receptor/kinase was examined by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis and gel filtration chromatography under denaturing or nondenaturing conditions. Pretreatment of 32 P-labeled insulin receptors with 50 mM DTT followed by gel filtration chromatography in 0.1% SDS demonstrated the dissociation of the α 2 β 2 insulin receptor complex (M/sub r/ 400,000) into the monomeric 95,000 β subunit. In contrast, pretreatment of the insulin receptors with 1-50 mM DTT followed by gel filtration chromatography in 0.1% Triton X-100 resulted in no apparent alteration in mobility compared to the untreated insulin receptors. Resolution of this complex by nonreducing SDS-polyacrylamide gel electrophoresis and autoradiography demonstrated the existence of the α 2 β 2 heterotetrameric complex with essentially no αβ heterodimeric or free monomeric β subunit species present. This suggests that the insulin receptor can reoxidize into the M/sub r/ 400,000 complex after the removal of DTT by gel filtration chromatography. To prevent reoxidation, the insulin receptors were pretreated with 50 mM DTT. Under the conditions the insulin receptors migrated as the M/sub r/ 400,000 α 2 β 2 complex. These results demonstrate that treatment of the insulin receptors with high concentrations of DTT, followed by removal of DTT by gel filtration, results in reoxidation of the reduced α 2 β 2 insulin receptor complex. Further, these results document that although the DTT stimulation of the insulin receptor/kinase does involve reduction of the insulin receptor subunits, it does not result in dissociation of the native α 2 β 2 insulin receptor subunit complex

  15. Involvement of the catalytic subunit of protein kinase A and of HA95 in pre-mRNA splicing

    International Nuclear Information System (INIS)

    Kvissel, Anne-Katrine; Orstavik, Sigurd; Eikvar, Sissel; Brede, Gaute; Jahnsen, Tore; Collas, Philippe; Akusjaervi, Goeran; Skalhegg, Bjorn Steen

    2007-01-01

    Protein kinase A (PKA) is a holoenzyme consisting of two catalytic (C) subunits bound to a regulatory (R) subunit dimer. Stimulation by cAMP dissociates the holoenzyme and causes translocation to the nucleus of a fraction of the C subunit. Apart from transcription regulation, little is known about the function of the C subunit in the nucleus. In the present report, we show that both Cα and Cβ are localized to spots in the mammalian nucleus. Double immunofluorescence analysis of splicing factor SC35 with the C subunit indicated that these spots are splicing factor compartments (SFCs). Using the E1A in vivo splicing assay, we found that catalytically active C subunits regulate alternative splicing and phosphorylate several members of the SR-protein family of splicing factors in vitro. Furthermore, nuclear C subunits co-localize with the C subunit-binding protein homologous to AKAP95, HA95. HA95 also regulates E1A alternative splicing in vivo, apparently through its N-terminal domain. Localization of the C subunit to SFCs and the E1A splicing pattern were unaffected by cAMP stimulation. Our findings demonstrate that the nuclear PKA C subunit co-locates with HA95 in SFCs and regulates pre-mRNA splicing, possibly through a cAMP-independent mechanism

  16. Fast and Slow Inhibition in the Visual Thalamus Is Influenced by Allocating GABAA Receptors with Different γ Subunits

    Directory of Open Access Journals (Sweden)

    Zhiwen Ye

    2017-04-01

    Full Text Available Cell-type specific differences in the kinetics of inhibitory postsynaptic conductance changes (IPSCs are believed to impact upon network dynamics throughout the brain. Much attention has focused on how GABAA receptor (GABAAR α and β subunit diversity will influence IPSC kinetics, but less is known about the influence of the γ subunit. We have examined whether GABAAR γ subunit heterogeneity influences IPSC properties in the thalamus. The γ2 subunit gene was deleted from GABAARs selectively in the dorsal lateral geniculate nucleus (dLGN. The removal of the γ2 subunit from the dLGN reduced the overall spontaneous IPSC (sIPSC frequency across all relay cells and produced an absence of IPSCs in a subset of relay neurons. The remaining slower IPSCs were both insensitive to diazepam and zinc indicating the absence of the γ2 subunit. Because these slower IPSCs were potentiated by methyl-6,7-dimethoxy-4-ethyl-β-carboline-3-carboxylate (DMCM, we propose these IPSCs involve γ1 subunit-containing GABAAR activation. Therefore, γ subunit heterogeneity appears to influence the kinetics of GABAAR-mediated synaptic transmission in the visual thalamus in a cell-selective manner. We suggest that activation of γ1 subunit-containing GABAARs give rise to slower IPSCs in general, while faster IPSCs tend to be mediated by γ2 subunit-containing GABAARs.

  17. Quantification of Transthyretin Kinetic Stability in Human Plasma Using Subunit Exchange

    Science.gov (United States)

    2015-01-01

    The transthyretin (TTR) amyloidoses are a group of degenerative diseases caused by TTR aggregation, requiring rate-limiting tetramer dissociation. Kinetic stabilization of TTR, by preferential binding of a drug to the native tetramer over the dissociative transition state, dramatically slows the progression of familial amyloid polyneuropathy. An established method for quantifying the kinetic stability of recombinant TTR tetramers in buffer is subunit exchange, in which tagged TTR homotetramers are added to untagged homotetramers at equal concentrations to measure the rate at which the subunits exchange. Herein, we report a subunit exchange method for quantifying the kinetic stability of endogenous TTR in human plasma. The subunit exchange reaction is initiated by the addition of a substoichiometric quantity of FLAG-tagged TTR homotetramers to endogenous TTR in plasma. Aliquots of the subunit exchange reaction, taken as a function of time, are then added to an excess of a fluorogenic small molecule, which immediately arrests further subunit exchange. After binding, the small molecule reacts with the TTR tetramers, rendering them fluorescent and detectable in human plasma after subsequent ion exchange chromatography. The ability to report on the extent of TTR kinetic stabilization resulting from treatment with oral tafamidis is important, especially for selection of the appropriate dose for patients carrying rare mutations. This method could also serve as a surrogate biomarker for the prediction of the clinical outcome. Subunit exchange was used to quantify the stabilization of WT TTR from senile systemic amyloidosis patients currently being treated with tafamidis (20 mg orally, once daily). TTR kinetic stability correlated with the tafamidis plasma concentration. PMID:24661308

  18. Immunodominant role of CCHA subunit of Concholepas hemocyanin is associated with unique biochemical properties.

    Science.gov (United States)

    Becker, María Inés; Fuentes, Alejandra; Del Campo, Miguel; Manubens, Augusto; Nova, Esteban; Oliva, Harold; Faunes, Fernando; Valenzuela, María Antonieta; Campos-Vallette, Marcelo; Aliaga, Alvaro; Ferreira, Jorge; De Ioannes, Alfredo E; De Ioannes, Pablo; Moltedo, Bruno

    2009-03-01

    Hemocyanin, the oxygen transporter metallo-glycoprotein from mollusks, shows strong relationship between its notable structural features and intrinsic immunomodulatory effects. Here we investigated the individual contribution of CCHA and CCHB subunits from Concholepas hemocyanin (CCH) to in vivo humoral immune response and their pre-clinical evaluation as immunotherapeutic agent in a mice bladder cancer model, in relation to their biochemical properties. To this end, subunits were purified and well characterized. Homogeneous subunits were obtained by anionic exchange chromatography, and its purity assessed by electrophoretic and immunochemical methods. While each CCH subunit contains eight functional units showing partial cross reaction, the vibrational spectral analysis showed several spectral differences, suggesting structural differences between them. In addition, we demonstrated differences in the carbohydrate content: CCHA had a 3.6% w/w sugar with both N- and O-linked moieties. In turn, CCHB had a 2.5% w/w sugar with N-linked, while O-linked moieties were nearly absent. Considering these differences, it was not possible to predict a priori whether the immunogenic and immunotherapeutic properties of subunits might be similar. Surprisingly, both subunits by itself induced a humoral response, and showed an antitumor effect in the bladder carcinoma cell line MBT-2. However, when immunologic parameters were analyzed, CCHA showed better efficiency than CCHB. No allergic reactions or any toxic effects were observed in mice treated with CCHA, sustaining its potential therapeutic use. Our study supports that CCHA subunit accounts for the most important features involved in the immunogenicity of CCH, such as better hydrophilicity and higher content of carbohydrates.

  19. Permeability transition in human mitochondria persists in the absence of peripheral stalk subunits of ATP synthase.

    Science.gov (United States)

    He, Jiuya; Carroll, Joe; Ding, Shujing; Fearnley, Ian M; Walker, John E

    2017-08-22

    The opening of a nonspecific channel, known as the permeability transition pore (PTP), in the inner membranes of mitochondria can be triggered by calcium ions, leading to swelling of the organelle, disruption of the inner membrane and ATP synthesis, and cell death. Pore opening can be inhibited by cyclosporin A mediated via cyclophilin D. It has been proposed that the pore is associated with the dimeric ATP synthase and the oligomycin sensitivity conferral protein (OSCP), a component of the enzyme's peripheral stalk, provides the site at which cyclophilin D interacts. Subunit b contributes a central α-helical structure to the peripheral stalk, extending from near the top of the enzyme's catalytic domain and crossing the membrane domain of the enzyme via two α-helices. We investigated the possible involvement of the subunit b and the OSCP in the PTP by generating clonal cells, HAP1-Δb and HAP1-ΔOSCP, lacking the membrane domain of subunit b or the OSCP, respectively, in which the corresponding genes, ATP5F1 and ATP5O , had been disrupted. Both cell lines preserve the characteristic properties of the PTP; therefore, the membrane domain of subunit b does not contribute to the PTP, and the OSCP does not provide the site of interaction with cyclophilin D. The membrane subunits ATP6, ATP8, and subunit c have been eliminated previously from possible participation in the PTP; thus, the only subunits of ATP synthase that could participate in pore formation are e, f, g, diabetes-associated protein in insulin-sensitive tissues (DAPIT), and the 6.8-kDa proteolipid.

  20. The biosynthesis and processing of high molecular weight precursors of soybean glycinin subunits.

    Science.gov (United States)

    Barton, K A; Thompson, J F; Madison, J T; Rosenthal, R; Jarvis, N P; Beachy, R N

    1982-06-10

    The predominant storage protein of soybean seed, glycinin, is composed of two heterogeneous classes of related subunits, the acidics (Mr approximately 38,000) and the basics (Mr approximately 22,000). Immunoreaction of polypeptides translated in vitro from isolated seed mRNA using antibodies prepared against either purified acidic or basic subunit groups precipitated precursor polypeptides of Mr = 60,000 to Mr = 63,000. High pressure liquid chromatography fingerprinting of trypsin-generated fragments from in vitro synthesized precursors showed fragments specific to both acidic and basic subunits. No mature acidic or basic subunits were detected in vitro translation reactions by either immunoprecipitation or high pressure liquid chromatography fingerprinting. Pulse-labeling of cotyledons growing in culture with [3H]glycine showed rapid accumulation of label in glycinin precursors of Mr = 59,000 to Mr = 62,000. Although in vivo synthesized precursors had slightly greater electrophoretic mobility than in vitro synthesized precursors, little label initially appeared in mature glycinin subunits. After several hours of continued cotyledon growth in absence of label, precursors were processed and label accumulated in both acidic and basic subunit groups. Recombinant plasmids were prepared by reverse transcription of soybean seed mRNA, and clones which encode glycinin precursors were identified by heteroduplex-hybridization of translatable messages. Northern blot analysis of seed mRNA shows the mRNA-encoding glycinin precursors to migrate at Mr = 0.71 X 10(6) on agarose gels, corresponding to approximately 2050 nucleotides. This is sufficiently large to encode a polypeptide consisting of both a glycinin acidic and basic subunit.

  1. Molecular mechanism of DNA recognition by the alpha subunit of the Oxytricha telomere binding protein.

    Science.gov (United States)

    Laporte, L; Benevides, J M; Thomas, G J

    1999-01-12

    Interactions between telomeric DNA and the alpha subunit of the heterodimeric telomere binding protein of Oxytricha nova have been probed by Raman spectroscopy, CD spectroscopy, and nondenaturing gel electrophoresis. Telomeric sequences investigated include the Oxytricha 3' overhang, d(T4G4)2, and the related sequence dT6(T4G4)2, which incorporates a 5'-thymidylate leader. Corresponding nontelomeric isomers, d(TG)8 and dT6(TG)8, have also been investigated. Both d(T4G4)2 and dT6(T4G4)2 form stable hairpins that contain Hoogsteen G.G base pairs [Laporte, L., and Thomas, G. J., Jr. (1998) J. Mol. Biol. 281, 261-270]. The alpha subunit binds specifically and stoichiometrically to the dT6(T4G4)2 hairpin and alters its secondary structure by inducing conformational changes in the 5'-thymidylate leader without extensive disruption of G.G base pairing. Conversely, binding of the alpha subunit to d(T4G4)2 eliminates G.G pairing and unfolds the hairpin. DNA unfolding is accompanied by conformational changes affecting both the backbone and dG residues, as evidenced by Raman and CD spectra. Interestingly, the alpha subunit also forms complexes with the nontelomeric isomers, d(TG)8 and dT6(TG)8, evidenced by altered electrophoretic mobility in nondenaturing gels; however, Raman and CD spectra of complexes of the alpha subunit with nontelomeric DNA suggest no significant changes in backbone or deoxynucleoside conformations. Similarly, the alpha subunit binds to but does not appreciably alter the secondary structure of duplex DNA. The present results show that while the alpha subunit has the capacity to bind to Watson-Crick and different non-Watson-Crick motifs, DNA refolding is specific to the Oxytricha telomeric hairpin and the retention of G.G pairing is specific to the telomeric sequence incorporating the 5' leading sequence. A model is proposed for alpha subunit binding to telomeric DNA, and the physiological role of the alpha subunit in telomere organization is discussed.

  2. The calcium channel β2 (CACNB2 subunit repertoire in teleosts

    Directory of Open Access Journals (Sweden)

    Mueller Rachel

    2008-04-01

    Full Text Available Abstract Background Cardiomyocyte contraction is initiated by influx of extracellular calcium through voltage-gated calcium channels. These oligomeric channels utilize auxiliary β subunits to chaperone the pore-forming α subunit to the plasma membrane, and to modulate channel electrophysiology 1. Several β subunit family members are detected by RT-PCR in the embryonic heart. Null mutations in mouse β2, but not in the other three β family members, are embryonic lethal at E10.5 due to defects in cardiac contractility 2. However, a drawback of the mouse model is that embryonic heart rhythm is difficult to study in live embryos due to their intra-uterine development. Moreover, phenotypes may be obscured by secondary effects of hypoxia. As a first step towards developing a model for contributions of β subunits to the onset of embryonic heart rhythm, we characterized the structure and expression of β2 subunits in zebrafish and other teleosts. Results Cloning of two zebrafish β2 subunit genes (β2.1 and β2.2 indicated they are membrane-associated guanylate kinase (MAGUK-family genes. Zebrafish β2 genes show high conservation with mammals within the SH3 and guanylate kinase domains that comprise the "core" of MAGUK proteins, but β2.2 is much more divergent in sequence than β2.1. Alternative splicing occurs at the N-terminus and within the internal HOOK domain. In both β2 genes, alternative short ATG-containing first exons are separated by some of the largest introns in the genome, suggesting that individual transcript variants could be subject to independent cis-regulatory control. In the Tetraodon nigrovidis and Fugu rubripes genomes, we identified single β2 subunit gene loci. Comparative analysis of the teleost and human β2 loci indicates that the short 5' exon sequences are highly conserved. A subset of 5' exons appear to be unique to teleost genomes, while others are shared with mammals. Alternative splicing is temporally and

  3. Functional Analysis of a Wheat AGPase Plastidial Small Subunit with a Truncated Transit Peptide

    Directory of Open Access Journals (Sweden)

    Yang Yang

    2017-03-01

    Full Text Available ADP-glucose pyrophosphorylase (AGPase, the key enzyme in starch synthesis, consists of two small subunits and two large subunits with cytosolic and plastidial isoforms. In our previous study, a cDNA sequence encoding the plastidial small subunit (TaAGPS1b of AGPase in grains of bread wheat (Triticum aestivum L. was isolated and the protein subunit encoded by this gene was characterized as a truncated transit peptide (about 50% shorter than those of other plant AGPS1bs. In the present study, TaAGPS1b was fused with green fluorescent protein (GFP in rice protoplast cells, and confocal fluorescence microscopy observations revealed that like other AGPS1b containing the normal transit peptide, TaAGPS1b-GFP was localized in chloroplasts. TaAGPS1b was further overexpressed in a Chinese bread wheat cultivar, and the transgenic wheat lines exhibited a significant increase in endosperm AGPase activities, starch contents, and grain weights. These suggested that TaAGPS1b subunit was targeted into plastids by its truncated transit peptide and it could play an important role in starch synthesis in bread wheat grains.

  4. Effect of adjuvants on responses to skin immunization by microneedles coated with influenza subunit vaccine.

    Directory of Open Access Journals (Sweden)

    William C Weldon

    Full Text Available Recent studies have demonstrated the effectiveness of vaccine delivery to the skin by vaccine-coated microneedles; however there is little information on the effects of adjuvants using this approach for vaccination. Here we investigate the use of TLR ligands as adjuvants with skin-based delivery of influenza subunit vaccine. BALB/c mice received 1 µg of monovalent H1N1 subunit vaccine alone or with 1 µg of imiquimod or poly(I:C individually or in combination via coated microneedle patches inserted into the skin. Poly(I:C adjuvanted subunit influenza vaccine induced similar antigen-specific immune responses compared to vaccine alone when delivered to the skin by microneedles. However, imiquimod-adjuvanted vaccine elicited higher levels of serum IgG2a antibodies and increased hemagglutination inhibition titers compared to vaccine alone, suggesting enhanced induction of functional antibodies. In addition, imiquimod-adjuvanted vaccine induced a robust IFN-γ cellular response. These responses correlated with improved protection compared to influenza subunit vaccine alone, as well as reduced viral replication and production of pro-inflammatory cytokines in the lungs. The finding that microneedle delivery of imiquimod with influenza subunit vaccine induces improved immune responses compared to vaccine alone supports the use of TLR7 ligands as adjuvants for skin-based influenza vaccines.

  5. Evaluation of peptide designing strategy against subunit reassociation in mucin 1: A steered molecular dynamics approach.

    Directory of Open Access Journals (Sweden)

    J Lesitha Jeeva Kumari

    Full Text Available Subunit reassociation in mucin 1, a breast cancer tumor marker, is reported as one of the critical factors for its cytoplasmic activation. Inhibition of its heterodimeric association would therefore result in loss of its function and alter disease progression. The present study aimed at evaluating peptide inhibitor designing strategies that may serve as antagonist against this receptor-ligand alliance. Several peptides and their derivatives were designed based on native residues, subunit interface, hydrogen bonding and secondary structure. Docking studies with the peptides were carried on the receptor subunit and their binding affinities were evaluated using steered molecular dynamics simulation and umbrella sampling. Our results showed that among all the different classes of peptides evaluated, the receptor based peptide showed the highest binding affinity. This result was concurrent with the experimental observation that the receptor-ligand alliance in mucin 1 is highly specific. Our results also show that peptide ligand against this subunit association is only stabilized through native residue inter-protein interaction irrespective of the peptide structure, peptide length and number of hydrogen bonds. Consistency in binding affinity, pull force and free energy barrier was observed with only the receptor derived peptides which resulted in favorable interprotein interactions at the interface. Several observations were made and discussed which will eventually lead to designing efficient peptide inhibitors against mucin 1 heterodimeric subunit reassociation.

  6. Radiation inactivation of glutamate dehydrogenase hexamer: lack of energy transfer between subunits

    International Nuclear Information System (INIS)

    Kempner, E.S.; Miller, J.H.

    1983-01-01

    The effects of ionizing radiation on glutamate dehydrogenase and on fluorescein isothiocyanate--tagged glutamate dehydrogenase were analyzed by target theory. Enzymatic activity, fluorescence, and the survival of the 56,000-dalton monomer subunit were determined on frozen samples irradiated at -135 degrees C and on lyophilized samples irradiated at either -135 degrees C or +30 degrees C. The effects of temperature were the same for all three parameters. Enzymatic activity was lost after small doses of high-energy electrons, whereas fluorescence and monomer subunits survived much larger doses of radiation. Target analysis revealed that the functional unit size for enzymatic activity was the hexamer, confirming both the earlier radiation study and conventional biochemical analyses. Target sizes obtained from fluorescence and subunit structure measurements were close to that of the monomer. These results indicate that the primary ionization caused by electron bombardment results in damage to a single polypeptide strand and that there is no massive transfer of radiation energy to other units in the hexamer. The large target size observed for enzymatic activity appears to be a structural requirement for the simultaneous presence of six intact subunits rather than the result of the spread of energy from the initial site to adjacent chains with consequent damage to other subunits

  7. Structural analysis of the α subunit of Na(+)/K(+) ATPase genes in invertebrates.

    Science.gov (United States)

    Thabet, Rahma; Rouault, J-D; Ayadi, Habib; Leignel, Vincent

    2016-01-01

    The Na(+)/K(+) ATPase is a ubiquitous pump coordinating the transport of Na(+) and K(+) across the membrane of cells and its role is fundamental to cellular functions. It is heteromer in eukaryotes including two or three subunits (α, β and γ which is specific to the vertebrates). The catalytic functions of the enzyme have been attributed to the α subunit. Several complete α protein sequences are available, but only few gene structures were characterized. We identified the genomic sequences coding the α-subunit of the Na(+)/K(+) ATPase, from the whole-genome shotgun contigs (WGS), NCBI Genomes (chromosome), Genomic Survey Sequences (GSS) and High Throughput Genomic Sequences (HTGS) databases across distinct phyla. One copy of the α subunit gene was found in Annelida, Arthropoda, Cnidaria, Echinodermata, Hemichordata, Mollusca, Placozoa, Porifera, Platyhelminthes, Urochordata, but the nematodes seem to possess 2 to 4 copies. The number of introns varied from 0 (Platyhelminthes) to 26 (Porifera); and their localization and length are also highly variable. Molecular phylogenies (Maximum Likelihood and Maximum Parsimony methods) showed some clusters constituted by (Chordata/(Echinodermata/Hemichordata)) or (Plathelminthes/(Annelida/Mollusca)) and a basal position for Porifera. These structural analyses increase our knowledge about the evolutionary events of the α subunit genes in the invertebrates. Copyright © 2016 Elsevier Inc. All rights reserved.

  8. Prokaryotic expression of the chicken xanthine oxidase (XOD) subunit and its localization in liver and kidney.

    Science.gov (United States)

    Lin, Huayuan; Chen, Yanjun; Huang, Qiqi; Guo, Xiaoquan; Liu, Ping; Liu, Weilian; Zhang, Caiying; Cao, Huabin; Hu, Guoliang

    2016-06-01

    Xanthine oxidase (XOD) is the members of the molybdenum hydroxylase flavoprotein family and it plays a vital role in the body's purine catabolism. In this study, we cloned the XOD 37kDa subunit protein by using RT-PCR and pMD-18-T clone vector based on the total RNA extracted from chicken liver. The cloning XOD subunit protein gene was ligated into the pET-32a to construct the recombinant plasmid pET-XOD. After the pET-XOD expression vector was transformed into host cells Rosetta (DE3), the recombinant XOD subunit proteins (54.8kDa) were successfully induced by isopropy1 β-d-thiogalactoside (IPTG). Rabbit antiserums were produced by using the purification of the recombinant XOD subunit protein as antigen. The titer of the antiserum was more than 1:102,400 determined by using ELISA. The result of Western blot demonstrated that the antiserum could specifically recognize the chicken liver XOD. Immunohistochemistry and immunofluorescence showed that the XOD mainly presented in the cytoplasm of chicken hepatocytes and proximal tubular epithelial cells. Our results indicated that the XOD subunit protein polyclonal antibody prepared by this method could be used for the further researches of the biological function of the XOD in the chicken. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Kinetic pathway of 40S ribosomal subunit recruitment to hepatitis C virus internal ribosome entry site.

    Science.gov (United States)

    Fuchs, Gabriele; Petrov, Alexey N; Marceau, Caleb D; Popov, Lauren M; Chen, Jin; O'Leary, Seán E; Wang, Richard; Carette, Jan E; Sarnow, Peter; Puglisi, Joseph D

    2015-01-13

    Translation initiation can occur by multiple pathways. To delineate these pathways by single-molecule methods, fluorescently labeled ribosomal subunits are required. Here, we labeled human 40S ribosomal subunits with a fluorescent SNAP-tag at ribosomal protein eS25 (RPS25). The resulting ribosomal subunits could be specifically labeled in living cells and in vitro. Using single-molecule Förster resonance energy transfer (FRET) between RPS25 and domain II of the hepatitis C virus (HCV) internal ribosome entry site (IRES), we measured the rates of 40S subunit arrival to the HCV IRES. Our data support a single-step model of HCV IRES recruitment to 40S subunits, irreversible on the initiation time scale. We furthermore demonstrated that after binding, the 40S:HCV IRES complex is conformationally dynamic, undergoing slow large-scale rearrangements. Addition of translation extracts suppresses these fluctuations, funneling the complex into a single conformation on the 80S assembly pathway. These findings show that 40S:HCV IRES complex formation is accompanied by dynamic conformational rearrangements that may be modulated by initiation factors.

  10. Structural insights into methyltransferase KsgA function in 30S ribosomal subunit biogenesis.

    Science.gov (United States)

    Boehringer, Daniel; O'Farrell, Heather C; Rife, Jason P; Ban, Nenad

    2012-03-23

    The assembly of the ribosomal subunits is facilitated by ribosome biogenesis factors. The universally conserved methyltransferase KsgA modifies two adjacent adenosine residues in the 3'-terminal helix 45 of the 16 S ribosomal RNA (rRNA). KsgA recognizes its substrate adenosine residues only in the context of a near mature 30S subunit and is required for the efficient processing of the rRNA termini during ribosome biogenesis. Here, we present the cryo-EM structure of KsgA bound to a nonmethylated 30S ribosomal subunit. The structure reveals that KsgA binds to the 30S platform with the catalytic N-terminal domain interacting with substrate adenosine residues in helix 45 and the C-terminal domain making extensive contacts to helix 27 and helix 24. KsgA excludes the penultimate rRNA helix 44 from adopting its position in the mature 30S subunit, blocking the formation of the decoding site and subunit joining. We suggest that the activation of methyltransferase activity and subsequent dissociation of KsgA control conformational changes in helix 44 required for final rRNA processing and translation initiation.

  11. Structural Insights into Methyltransferase KsgA Function in 30S Ribosomal Subunit Biogenesis*

    Science.gov (United States)

    Boehringer, Daniel; O'Farrell, Heather C.; Rife, Jason P.; Ban, Nenad

    2012-01-01

    The assembly of the ribosomal subunits is facilitated by ribosome biogenesis factors. The universally conserved methyltransferase KsgA modifies two adjacent adenosine residues in the 3′-terminal helix 45 of the 16 S ribosomal RNA (rRNA). KsgA recognizes its substrate adenosine residues only in the context of a near mature 30S subunit and is required for the efficient processing of the rRNA termini during ribosome biogenesis. Here, we present the cryo-EM structure of KsgA bound to a nonmethylated 30S ribosomal subunit. The structure reveals that KsgA binds to the 30S platform with the catalytic N-terminal domain interacting with substrate adenosine residues in helix 45 and the C-terminal domain making extensive contacts to helix 27 and helix 24. KsgA excludes the penultimate rRNA helix 44 from adopting its position in the mature 30S subunit, blocking the formation of the decoding site and subunit joining. We suggest that the activation of methyltransferase activity and subsequent dissociation of KsgA control conformational changes in helix 44 required for final rRNA processing and translation initiation. PMID:22308031

  12. The complete structure of the large subunit of the mammalian mitochondrial ribosome.

    Science.gov (United States)

    Greber, Basil J; Boehringer, Daniel; Leibundgut, Marc; Bieri, Philipp; Leitner, Alexander; Schmitz, Nikolaus; Aebersold, Ruedi; Ban, Nenad

    2014-11-13

    Mitochondrial ribosomes (mitoribosomes) are extensively modified ribosomes of bacterial descent specialized for the synthesis and insertion of membrane proteins that are critical for energy conversion and ATP production inside mitochondria. Mammalian mitoribosomes, which comprise 39S and 28S subunits, have diverged markedly from the bacterial ribosomes from which they are derived, rendering them unique compared to bacterial, eukaryotic cytosolic and fungal mitochondrial ribosomes. We have previously determined at 4.9 Å resolution the architecture of the porcine (Sus scrofa) 39S subunit, which is highly homologous to the human mitoribosomal large subunit. Here we present the complete atomic structure of the porcine 39S large mitoribosomal subunit determined in the context of a stalled translating mitoribosome at 3.4 Å resolution by cryo-electron microscopy and chemical crosslinking/mass spectrometry. The structure reveals the locations and the detailed folds of 50 mitoribosomal proteins, shows the highly conserved mitoribosomal peptidyl transferase active site in complex with its substrate transfer RNAs, and defines the path of the nascent chain in mammalian mitoribosomes along their idiosyncratic exit tunnel. Furthermore, we present evidence that a mitochondrial tRNA has become an integral component of the central protuberance of the 39S subunit where it architecturally substitutes for the absence of the 5S ribosomal RNA, a ubiquitous component of all cytoplasmic ribosomes.

  13. In Search of Small Molecule Inhibitors Targeting the Flexible CK2 Subunit Interface

    Directory of Open Access Journals (Sweden)

    Benoît Bestgen

    2017-02-01

    Full Text Available Protein kinase CK2 is a tetrameric holoenzyme composed of two catalytic (α and/or α’ subunits and two regulatory (β subunits. Crystallographic data paired with fluorescence imaging techniques have suggested that the formation of the CK2 holoenzyme complex within cells is a dynamic process. Although the monomeric CK2α subunit is endowed with a constitutive catalytic activity, many of the plethora of CK2 substrates are exclusively phosphorylated by the CK2 holoenzyme. This means that the spatial and high affinity interaction between CK2α and CK2β subunits is critically important and that its disruption may provide a powerful and selective way to block the phosphorylation of substrates requiring the presence of CK2β. In search of compounds inhibiting this critical protein–protein interaction, we previously designed an active cyclic peptide (Pc derived from the CK2β carboxy-terminal domain that can efficiently antagonize the CK2 subunit interaction. To understand the functional significance of this interaction, we generated cell-permeable versions of Pc, exploring its molecular mechanisms of action and the perturbations of the signaling pathways that it induces in intact cells. The identification of small molecules inhibitors of this critical interaction may represent the first-choice approach to manipulate CK2 in an unconventional way.

  14. Distribution of protein and RNA in the 30S ribosomal subunit

    International Nuclear Information System (INIS)

    Ramakrishnan, V.

    1986-01-01

    In Escherichia coli, the small ribosomal subunit has a sedimentation coefficient of 30S, and consists of a 16S RNA molecule of 1541 nucleotides complexed with 21 proteins. Over the last few years, a controversy has emerged regarding the spatial distribution of RNA and protein in the 30S subunit. Contrast variation with neutron scattering was used to suggest that the RNA was located in a central core of the subunit and the proteins mainly in the periphery, with virtually no separation between the centers of mass of protein and RNA. However, these findings are incompatible with the results of efforts to locate individual ribosomal proteins by immune electron microscopy and triangulation with interprotein distance measurements. The conflict between these two views is resolved in this report of small-angle neutron scattering measurements on 30S subunits with and without protein S1, and on subunits reconstituted from deuterated 16S RNA and unlabeled proteins. The results show that (i) the proteins and RNA are intermingled, with neither component dominating at the core or the periphery, and (ii) the spatial distribution of protein and RNA is asymmetrical, with a separation between their centers of mass of about 25 angstroms

  15. APC16 is a conserved subunit of the anaphase-promoting complex/cyclosome.

    Science.gov (United States)

    Kops, Geert J P L; van der Voet, Monique; van der Voet, Moniek; Manak, Michael S; van Osch, Maria H J; Naini, Said M; Brear, Andrea; McLeod, Ian X; Hentschel, Dirk M; Yates, John R; van den Heuvel, Sander; Shah, Jagesh V

    2010-05-15

    Error-free chromosome segregation depends on timely activation of the multi-subunit E3 ubiquitin ligase APC/C. Activation of the APC/C initiates chromosome segregation and mitotic exit by targeting critical cell-cycle regulators for destruction. The APC/C is the principle target of the mitotic checkpoint, which prevents segregation while chromosomes are unattached to spindle microtubules. We now report the identification and characterization of APC16, a conserved subunit of the APC/C. APC16 was found in association with tandem-affinity-purified mitotic checkpoint complex protein complexes. APC16 is a bona fide subunit of human APC/C: it is present in APC/C complexes throughout the cell cycle, the phenotype of APC16-depleted cells copies depletion of other APC/C subunits, and APC16 is important for APC/C activity towards mitotic substrates. APC16 sequence homologues can be identified in metazoans, but not fungi, by four conserved primary sequence stretches. We provide evidence that the C. elegans gene K10D2.4 and the D. rerio gene zgc:110659 are functional equivalents of human APC16. Our findings show that APC/C is composed of previously undescribed subunits, and raise the question of why metazoan APC/C is molecularly different from unicellular APC/C.

  16. Goniometer-based femtosecond X-ray diffraction of mutant 30S ribosomal subunit crystals

    Directory of Open Access Journals (Sweden)

    E. Han Dao

    2015-07-01

    Full Text Available In this work, we collected radiation-damage-free data from a set of cryo-cooled crystals for a novel 30S ribosomal subunit mutant using goniometer-based femtosecond crystallography. Crystal quality assessment for these samples was conducted at the X-ray Pump Probe end-station of the Linac Coherent Light Source (LCLS using recently introduced goniometer-based instrumentation. These 30S subunit crystals were genetically engineered to omit a 26-residue protein, Thx, which is present in the wild-type Thermus thermophilus 30S ribosomal subunit. We are primarily interested in elucidating the contribution of this ribosomal protein to the overall 30S subunit structure. To assess the viability of this study, femtosecond X-ray diffraction patterns from these crystals were recorded at the LCLS during a protein crystal screening beam time. During our data collection, we successfully observed diffraction from these difficult-to-grow 30S ribosomal subunit crystals. Most of our crystals were found to diffract to low resolution, while one crystal diffracted to 3.2 Å resolution. These data suggest the feasibility of pursuing high-resolution data collection as well as the need to improve sample preparation and handling in order to collect a complete radiation-damage-free data set using an X-ray Free Electron Laser.

  17. Goniometer-based femtosecond X-ray diffraction of mutant 30S ribosomal subunit crystals.

    Science.gov (United States)

    Dao, E Han; Sierra, Raymond G; Laksmono, Hartawan; Lemke, Henrik T; Alonso-Mori, Roberto; Coey, Aaron; Larsen, Kevin; Baxter, Elizabeth L; Cohen, Aina E; Soltis, S Michael; DeMirci, Hasan

    2015-07-01

    In this work, we collected radiation-damage-free data from a set of cryo-cooled crystals for a novel 30S ribosomal subunit mutant using goniometer-based femtosecond crystallography. Crystal quality assessment for these samples was conducted at the X-ray Pump Probe end-station of the Linac Coherent Light Source (LCLS) using recently introduced goniometer-based instrumentation. These 30S subunit crystals were genetically engineered to omit a 26-residue protein, Thx, which is present in the wild-type Thermus thermophilus 30S ribosomal subunit. We are primarily interested in elucidating the contribution of this ribosomal protein to the overall 30S subunit structure. To assess the viability of this study, femtosecond X-ray diffraction patterns from these crystals were recorded at the LCLS during a protein crystal screening beam time. During our data collection, we successfully observed diffraction from these difficult-to-grow 30S ribosomal subunit crystals. Most of our crystals were found to diffract to low resolution, while one crystal diffracted to 3.2 Å resolution. These data suggest the feasibility of pursuing high-resolution data collection as well as the need to improve sample preparation and handling in order to collect a complete radiation-damage-free data set using an X-ray Free Electron Laser.

  18. Molecular investigations of BK(Ca) channels and the modulatory beta-subunits in porcine basilar and middle cerebral arteries

    DEFF Research Database (Denmark)

    Johansson, Helle Wulf; Hay-Schmidt, Anders; Poulsen, Asser Nyander

    2009-01-01

    arteries using reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time PCR. Western blotting was used to detect immunoreactivity for the porcine BK(Ca) channel alpha-subunit and beta-subunit proteins. The BK(Ca) channel alpha-subunit RNA and protein distribution patterns were......Large conductance calcium-activated potassium (BK(Ca)) channels are fundamental in the regulation of cerebral vascular basal tone. We investigated the expression of the mRNA transcripts for the BK(Ca) channel and its modulatory beta-subunits (beta1-beta4) in porcine basilar and middle cerebral...... visualized using in situ hybridization and immunofluorescence studies, respectively. The study verified that the BK(Ca) channel alpha-subunit is located to smooth muscle cells of porcine basilar and middle cerebral arteries. The mRNA transcript for beta1-, beta2- and beta4-subunit were shown by RT...

  19. Cloning and expression of the human N-methyl-D-aspartate receptor subunit NR3A

    DEFF Research Database (Denmark)

    Eriksson, Maria; Nilsson, Anna; Froelich-Fabre, Susanne

    2002-01-01

    Native N-methyl-D-aspartate (NMDA) receptors are heteromeric assemblies of four or five subunits. The NMDA receptor subunits, NR1, NR2A, NR2B, NR2C, and NR2D have been cloned in several species, including man. The NR3A subunit, which in rodents is predominantly expressed during early development......, seems to function by reducing the NMDA receptor response. The human homologue to the rat NR3A, however, had not been cloned. In order to study the functions of the human NR3A (hNR3A), we have cloned and sequenced the hNR3A. It was found to share 88% of the DNA sequence with the rat gene, corresponding...

  20. Unique Roles of the Non-identical MCM Subunits in DNA Replication Licensing.

    Science.gov (United States)

    Zhai, Yuanliang; Li, Ningning; Jiang, Hanlun; Huang, Xuhui; Gao, Ning; Tye, Bik Kwoon

    2017-07-20

    A family of six homologous subunits, Mcm2, -3, -4, -5, -6, and -7, each with its own unique features, forms the catalytic core of the eukaryotic replicative helicase. The necessity of six similar but non-identical subunits has been a mystery since its initial discovery. Recent cryo-EM structures of the Mcm2-7 (MCM) double hexamer, its precursors, and the origin recognition complex (ORC)-Cdc6-Cdt1-Mcm2-7 (OCCM) intermediate showed that each of these subunits plays a distinct role in orchestrating the assembly of the pre-replication complex (pre-RC) by ORC-Cdc6 and Cdt1. Copyright © 2017 Elsevier Inc. All rights reserved.