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Sample records for ancillary pilin subunit

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  4. Fundamental role for the KCNE4 ancillary subunit in Kv7.4 regulation of arterial tone

    DEFF Research Database (Denmark)

    Jepps, Thomas A; Carr, Georgina; Lundegaard, Pia R

    2015-01-01

    tone by Kv7 channels. In HEK cells expressing Kv7.4, co-expression of KCNE4 increased the membrane expression of Kv7.4 and significantly altered Kv7.4 current properties. Quantitative PCR analysis of different rat arteries found that the KCNE4 isoform predominated and proximity ligation experiments...... leads to reduced Kv7.4 membrane abundance, a depolarized membrane potential and an augmented response to vasoconstrictors. KCNE4 is a key regulator of the function and expression of Kv7.4 in vascular smooth muscle. ABSTRACT: The KCNE ancillary subunits (KCNE1-5) significantly alter the expression...... to reduced Kv7.4 membrane abundance, a depolarized membrane potential and an augmented response to vasoconstrictors. The present study is the first to demonstrate an integral role of KCNE4 in regulating the function and expression of Kv7.4 in vascular smooth muscle....

  5. 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

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

    Directory of Open Access Journals (Sweden)

    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

  7. The type IV pilin of Burkholderia mallei is highly immunogenic but fails to protect against lethal aerosol challenge in a murine model.

    Science.gov (United States)

    Fernandes, Paula J; Guo, Qin; Waag, David M; Donnenberg, Michael S

    2007-06-01

    Burkholderia mallei is the cause of glanders and a proven biological weapon. We identified and purified the type IV pilin protein of this organism to study its potential as a subunit vaccine. We found that purified pilin was highly immunogenic. Furthermore, mice infected via sublethal aerosol challenge developed significant increases in titers of antibody against the pilin, suggesting that it is expressed in vivo. Nevertheless, we found no evidence that high-titer antipilin antisera provided passive protection against a sublethal or lethal aerosol challenge and no evidence of protection afforded by active immunization with purified pilin. These results contrast with the utility of type IV pilin subunit vaccines against other infectious diseases and highlight the need for further efforts to identify protective responses against this pathogen.

  8. 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

    2015-01-01

    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. PMID:26324721

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

    Science.gov (United States)

    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. MOLECULAR DOCKING AND DYNAMICS STUDIES ON THE PROTEIN-PROTEIN INTERACTIONS OF ELECTRICALLY ACTIVE PILIN NANOWIRES OF GEOBACTER SULFURREDUCENS.

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    D. Jeya Sundara Sharmila1 *

    2017-06-01

    Full Text Available Molecular interactions are key aspects in biological recognitions applicable in nano/micro systems. Bacterial nanowires are pilus filament based structures that can conduct electrons. The transport of electron is proposed to be facilitated by filamentous fibers made up of polymeric assemblies of proteins called pilin. Geobacter sulfurreducens is capable of delivering electrons through extracellular electron transport (EET by employing conductive nanowires, which are pilin proteins composed of type IV subunit PilA. Protein-protein interactions play an important role in the stabilization of the pilin nanowire assembly complex and it contains transmembrane (TM domain. In current study, protein-protein docking and multiple molecular dynamic (MD simulations were performed to understand the binding mode of pilin nanowires. The MD result explains the conformational behavior and folding of pilin nanowires in water environment in different time scale duration 20, 5, 5, 10 and 20ns (total of 60ns. Direct hydrogen bonds and water mediated hydrogen bonds that play a crucial role during the simulation were investigated. The conformational state, folding, end-toend distance profile and hydrogen bonding behavior had indicated that the Geobacter sulfurreducens pilin nanowires have electrical conductivity properties.

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

    International Nuclear Information System (INIS)

    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°

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

  14. Type IV pilins regulate their own expression via direct intramembrane interactions with the sensor kinase PilS.

    Science.gov (United States)

    Kilmury, Sara L N; Burrows, Lori L

    2016-05-24

    Type IV pili are important virulence factors for many pathogens, including Pseudomonas aeruginosa Transcription of the major pilin gene-pilA-is controlled by the PilS-PilR two-component system in response to unknown signals. The absence of a periplasmic sensing domain suggested that PilS may sense an intramembrane signal, possibly PilA. We suggest that direct interactions between PilA and PilS in the inner membrane reduce pilA transcription when PilA levels are high. Overexpression in trans of PilA proteins with diverse and/or truncated C termini decreased native pilA transcription, suggesting that the highly conserved N terminus of PilA was the regulatory signal. Point mutations in PilA or PilS that disrupted their interaction prevented autoregulation of pilA transcription. A subset of PilA point mutants retained the ability to interact with PilS but could no longer decrease pilA transcription, suggesting that interaction between the pilin and sensor kinase is necessary but not sufficient for pilA autoregulation. Furthermore, PilS's phosphatase motif was required for the autoregulation of pilA transcription, suggesting that under conditions where PilA is abundant, the PilA-PilS interaction promotes PilR dephosphorylation and thus down-regulation of further pilA transcription. These data reveal a clever bacterial inventory control strategy in which the major subunit of an important P. aeruginosa virulence factor controls its own expression.

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

  18. Implementations of PI-line based FBP and BPF algorithms on GPGPU

    Energy Technology Data Exchange (ETDEWEB)

    Shen, Le [Tsinghua Univ., Beijing (China). Dept. of Engineering Physics; Xing, Yuxiang [Tsinghua Univ., Beijing (China). Dept. of Engineering Physics; Ministry of Education, Beijing (China). Key Lab. of Particle and Radiation Imaging

    2011-07-01

    Exact reconstruction is under the spotlight in cone beam CT. Katsevich put forward the first exact inversion formula for helical cone beam CT, which belongs to FBP type. Also, Pan Xiaochuan's group proposed another PI-line based exact reconstruction algorithm of BPF type. These two exact reconstruction algorithms and their derivative forms have been widely studied. In this paper, we present a different way of selecting PI-line segments appropriate for both Katsevich's FBP and Pan Xiaochuan's BPF algorithms. As 3D reconstruction contributes to massive computations and takes long time, people have made efforts to speed up the algorithms with the help of multi-core CPUs and GPGPU (General Purpose Graphics Processing Unit). In this paper, we also presents implementations for these two algorithms on GPGPU using an innovative way of selecting PI-line segments. Acceleration techniques and implementations are addressed in detail. The methods are tested on the Shepp-Logan phantom. Compared with our CPU's implementations, the accelerated algorithms on GPGPU are tens to hundreds times faster. (orig.)

  19. 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.

  20. 18 CFR 35.40 - Ancillary services.

    Science.gov (United States)

    2010-04-01

    ... 18 Conservation of Power and Water Resources 1 2010-04-01 2010-04-01 false Ancillary services. 35.40 Section 35.40 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION... Sales of Electric Energy, Capacity and Ancillary Services at Market-Based Rates § 35.40 Ancillary...

  1. In vitro reconstitution of sortase-catalyzed pilus polymerization reveals structural elements involved in pilin cross-linking.

    Science.gov (United States)

    Chang, Chungyu; Amer, Brendan R; Osipiuk, Jerzy; McConnell, Scott A; Huang, I-Hsiu; Hsieh, Van; Fu, Janine; Nguyen, Hong H; Muroski, John; Flores, Erika; Ogorzalek Loo, Rachel R; Loo, Joseph A; Putkey, John A; Joachimiak, Andrzej; Das, Asis; Clubb, Robert T; Ton-That, Hung

    2018-06-12

    Covalently cross-linked pilus polymers displayed on the cell surface of Gram-positive bacteria are assembled by class C sortase enzymes. These pilus-specific transpeptidases located on the bacterial membrane catalyze a two-step protein ligation reaction, first cleaving the LPXTG motif of one pilin protomer to form an acyl-enzyme intermediate and then joining the terminal Thr to the nucleophilic Lys residue residing within the pilin motif of another pilin protomer. To date, the determinants of class C enzymes that uniquely enable them to construct pili remain unknown. Here, informed by high-resolution crystal structures of corynebacterial pilus-specific sortase (SrtA) and utilizing a structural variant of the enzyme (SrtA 2M ), whose catalytic pocket has been unmasked by activating mutations, we successfully reconstituted in vitro polymerization of the cognate major pilin (SpaA). Mass spectrometry, electron microscopy, and biochemical experiments authenticated that SrtA 2M synthesizes pilus fibers with correct Lys-Thr isopeptide bonds linking individual pilins via a thioacyl intermediate. Structural modeling of the SpaA-SrtA-SpaA polymerization intermediate depicts SrtA 2M sandwiched between the N- and C-terminal domains of SpaA harboring the reactive pilin and LPXTG motifs, respectively. Remarkably, the model uncovered a conserved TP(Y/L)XIN(S/T)H signature sequence following the catalytic Cys, in which the alanine substitutions abrogated cross-linking activity but not cleavage of LPXTG. These insights and our evidence that SrtA 2M can terminate pilus polymerization by joining the terminal pilin SpaB to SpaA and catalyze ligation of isolated SpaA domains in vitro provide a facile and versatile platform for protein engineering and bio-conjugation that has major implications for biotechnology.

  2. 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.

  3. 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.

  4. 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

  5. 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

  6. Structural conservation, variability, and immunogenicity of the T6 backbone pilin of serotype M6 Streptococcus pyogenes.

    Science.gov (United States)

    Young, Paul G; Moreland, Nicole J; Loh, Jacelyn M; Bell, Anita; Atatoa Carr, Polly; Proft, Thomas; Baker, Edward N

    2014-07-01

    Group A streptococcus (GAS; Streptococcus pyogenes) is a Gram-positive human pathogen that causes a broad range of diseases ranging from acute pharyngitis to the poststreptococcal sequelae of acute rheumatic fever. GAS pili are highly diverse, long protein polymers that extend from the cell surface. They have multiple roles in infection and are promising candidates for vaccine development. This study describes the structure of the T6 backbone pilin (BP; Lancefield T-antigen) from the important M6 serotype. The structure reveals a modular arrangement of three tandem immunoglobulin-like domains, two with internal isopeptide bonds. The T6 pilin lysine, essential for polymerization, is located in a novel VAKS motif that is structurally homologous to the canonical YPKN pilin lysine in other three- and four-domain Gram-positive pilins. The T6 structure also highlights a conserved pilin core whose surface is decorated with highly variable loops and extensions. Comparison to other Gram-positive BPs shows that many of the largest variable extensions are found in conserved locations. Studies with sera from patients diagnosed with GAS-associated acute rheumatic fever showed that each of the three T6 domains, and the largest of the variable extensions (V8), are targeted by IgG during infection in vivo. Although the GAS BP show large variations in size and sequence, the modular nature of the pilus proteins revealed by the T6 structure may aid the future design of a pilus-based vaccine. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  7. Mucosal adhesion properties of the probiotic Lactobacillus rhamnosus GG SpaCBA and SpaFED pilin subunits

    NARCIS (Netherlands)

    Ossowski, von I.; Reunanen, J.; Satokari, R.M.; Vesterlund, S.; Kankainen, M.; Huhtinen, H.; Tynkkynen, S.; Salminen, S.; Vos, de W.M.; Palva, A.

    2010-01-01

    Lactobacillus rhamnosus GG is a well-established Gram-positive probiotic strain, whose health-benefiting properties are in part dependent upon a prolonged residency in the gastrointestinal tract and likely dictated by an adherence to the intestinal mucosa. Previously, we identified two pilus gene

  8. 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

  9. Type IV pilins regulate their own expression via direct intramembrane interactions with the sensor kinase PilS

    OpenAIRE

    Kilmury, Sara L. N.; Burrows, Lori L.

    2016-01-01

    Although two-component systems are a ubiquitous means of rapid bacterial adaptation to changing environments, identification of the specific signals detected by sensor kinases can be challenging. Also, little is known about the diverse, poorly characterized family of sensor kinases that detect intramembrane signals. We show that the major type IV pilin, PilA, is an inhibitory intramembrane ligand for the PilS sensor kinase that controls pilA expression and we characterize the mechanism of sig...

  10. 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

  11. Reconsidering the measurement of ancillary service performance.

    Science.gov (United States)

    Griffin, D T; Rauscher, J A

    1987-08-01

    Prospective payment reimbursement systems have forced hospitals to review their costs more carefully. The result of the increased emphasis on costs is that many hospitals use costs, rather than margin, to judge the performance of ancillary services. However, arbitrary selection of performance measures for ancillary services can result in managerial decisions contrary to hospital objectives. Managerial accounting systems provide models which assist in the development of performance measures for ancillary services. Selection of appropriate performance measures provides managers with the incentive to pursue goals congruent with those of the hospital overall. This article reviews the design and implementation of managerial accounting systems, and considers the impact of prospective payment systems and proposed changes in capital reimbursement on this process.

  12. 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.

  13. 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....

  14. Creating competitive markets for ancillary services

    Energy Technology Data Exchange (ETDEWEB)

    Hirst, E.; Kirby, B.

    1997-10-01

    This report describes the structure of, and results from, a spreadsheet model. The model simulates markets for seven services: losses, regulation, spinning reserve, supplemental reserve,load following, energy imbalance, and voltage support. For completeness, the model also calculates costs for system control, although this service will continue to be provided solely the the system operator under cost-based prices. This computer model demonstrated the likely complexity of markets for energy and ancillary services. This complexity arises because these markets are highly interdependent. Because these markets are interactive, the costs and therefore the prices of these services will vary considerably as functions of system load and the current spot price of energy. The price of ancillary services in aggregate is highly correlated with the price of energy. A base-case utility was developed for use with the model and included in the report.

  15. 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.

  16. 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.

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

    International Nuclear Information System (INIS)

    Krishnan, Vengadesan; Dwivedi, Prabhat; Kim, Brandon J.; Samal, Alexandra; Macon, Kevin; Ma, Xin; Mishra, Arunima; Doran, Kelly S.; Ton-That, Hung; Narayana, Sthanam V. L.

    2013-01-01

    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

  18. 42 CFR 414.40 - Coding and ancillary policies.

    Science.gov (United States)

    2010-10-01

    ... 42 Public Health 3 2010-10-01 2010-10-01 false Coding and ancillary policies. 414.40 Section 414.40 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES... Practitioners § 414.40 Coding and ancillary policies. (a) General rule. CMS establishes uniform national...

  19. 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...

  20. 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...

  1. 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...

  2. 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.

  3. 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. PMID:27070897

  4. 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

  5. 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....

  6. 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.

  7. 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.

  8. 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.

  9. New developments in segment ancillary logic for FASTBUS

    International Nuclear Information System (INIS)

    Walz, H.V.; Bertolucci, B.

    1983-01-01

    Segment Ancillary Logic hardware for FASTBUS systems provides logical functions required in common by all devices attached to a segment. It controls the execution of arbitration cycles, and geographical address cycles, and generates the system handshake responses for broadcast operations. The mandatory requirements for Segment Ancillary Logic in the FASTBUS specifications are reviewed. A detailed implementation based on ECL logic is described, and the hardware to be used on an ECL cable segment for an experimental FASTBUS system at SLAC is shown

  10. An analysis of ongoing trends in airline ancillary revenues

    OpenAIRE

    Warnock-Smith, David; O'Connell, John F.; Maleki, Mahnaz

    2017-01-01

    This paper examines the performance of the two core classifications of airline ancillary revenues, which are unbundled products and commission based income. It also investigates the willingness of passengers to pay (WTP) for these services together with what type of ancillary items are acceptable at a particular price point. The study found that passengers value a narrow range of perceived ‘necessity’ products and services such as food and drink, checked baggage and seat assignment as opposed...

  11. 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.

  12. Against the odds? De novo structure determination of a pilin with two cysteine residues by sulfur SAD.

    Science.gov (United States)

    Gorgel, Manuela; Bøggild, Andreas; Ulstrup, Jakob Jensen; Weiss, Manfred S; Müller, Uwe; Nissen, Poul; Boesen, Thomas

    2015-05-01

    Exploiting the anomalous signal of the intrinsic S atoms to phase a protein structure is advantageous, as ideally only a single well diffracting native crystal is required. However, sulfur is a weak anomalous scatterer at the typical wavelengths used for X-ray diffraction experiments, and therefore sulfur SAD data sets need to be recorded with a high multiplicity. In this study, the structure of a small pilin protein was determined by sulfur SAD despite several obstacles such as a low anomalous signal (a theoretical Bijvoet ratio of 0.9% at a wavelength of 1.8 Å), radiation damage-induced reduction of the cysteines and a multiplicity of only 5.5. The anomalous signal was improved by merging three data sets from different volumes of a single crystal, yielding a multiplicity of 17.5, and a sodium ion was added to the substructure of anomalous scatterers. In general, all data sets were balanced around the threshold values for a successful phasing strategy. In addition, a collection of statistics on structures from the PDB that were solved by sulfur SAD are presented and compared with the data. Looking at the quality indicator R(anom)/R(p.i.m.), an inconsistency in the documentation of the anomalous R factor is noted and reported.

  13. The evolving design of RTO ancillary service markets

    International Nuclear Information System (INIS)

    Isemonger, Alan G.

    2009-01-01

    Although the markets for ancillary services at the North American Independent System Operators are often structured in quite different ways there is an emerging set of core design elements that represent a rough consensus as to what the optimal design configuration for ancillary services should be, albeit with some regional variation. This paper looks back at how the design of ancillary services markets has recently evolved to put this development in context. Thereafter it examines the methods by which ancillary services are procured by highlighting the procurement practices at a number of different Independent System Operators, principally those in California, New York, New England, Texas and the PJM Interconnection, in an attempt to tease out the remaining reasons why the ancillary service markets are still so different. This is important as there are many innovations that are not rooted in regional differences but reflect genuine technical advances and economic efficiency gains and can be replicated across other ISOs to produce more efficient designs, greater reliability and lower costs. (author)

  14. 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.

  15. 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.

  16. 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....

  17. Reduced risks and improved economic operation of ancillary services

    NARCIS (Netherlands)

    Bosch, van den P.P.J.; Jokic, A.; Hermans, R.M.; Frunt, J.; Kling, W.L.; Nobel, Frank; Boonekamp, P.; Boer, de W.W.

    2010-01-01

    Present arrangements and regulation for ancillary services for power balance in power systems cannot cope with future developments in power systems as the participants do not receive proper incentives for required behaviour. This paper analyzes the consequences of current arrangements for system

  18. 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...

  19. 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.

  20. 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

  1. 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...... delivery. Consumers' involvement requires new entities and infrastructure. A so-called aggregator has been introduced as a new player to manage the services that are offered by the consumption units. This paper describes an industrial scale experimental setup for evaluating a particular type of aggregator....... The aggregator aims to provide a distribution grid service from industrial thermal loads through a direct control policy. Our specific case studies are a supermarket refrigeration system and an HVac chiller in conjunction with an ice storage, which are virtually connected to the aggregator. Practical results...

  2. An examination of ongoing trends in airline ancillary revenues

    OpenAIRE

    Warnock-Smith, David; O'Connell, John Frankie; Maleki, Mahnaz

    2015-01-01

    The airline industry seems permanently embedded in producing thin margins and continuously combatting downward pressure on yields. To perpetuate the problem, the industry remains eclipsed with high cost structures and low barriers to entry. However, a new sizzling concept continues to counterbalance these effects in the form of ancillary revenues. Globally, these revenues have increased by 121% from 2010 to 2014 – and the trend is set to continue as carriers are quickly implementing structura...

  3. 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.

  4. 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

  5. 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

  6. The Role of Chaperone-subunit Usher Domain Interactions in the Mechanism of Bacterial Pilus Biogenesis Revealed by ESI-MS*

    Science.gov (United States)

    Morrissey, Bethny; Leney, Aneika C.; Toste Rêgo, Ana; Phan, Gilles; Allen, William J.; Verger, Denis; Waksman, Gabriel; Ashcroft, Alison E.; Radford, Sheena E.

    2012-01-01

    The PapC usher is a β-barrel outer membrane protein essential for assembly and secretion of P pili that are required for adhesion of pathogenic E. coli, which cause the development of pyelonephritis. Multiple protein subunits form the P pilus, the highly specific assembly of which is coordinated by the usher. Despite a wealth of structural knowledge, how the usher catalyzes subunit polymerization and orchestrates a correct and functional order of subunit assembly remain unclear. Here, the ability of the soluble N-terminal (UsherN), C-terminal (UsherC2), and Plug (UsherP) domains of the usher to bind different chaperone-subunit (PapDPapX) complexes is investigated using noncovalent electrospray ionization mass spectrometry. The results reveal that each usher domain is able to bind all six PapDPapX complexes, consistent with an active role of all three usher domains in pilus biogenesis. Using collision induced dissociation, combined with competition binding experiments and dissection of the adhesin subunit, PapG, into separate pilin and adhesin domains, the results reveal why PapG has a uniquely high affinity for the usher, which is consistent with this subunit always being displayed at the pilus tip. In addition, we show how the different soluble usher domains cooperate to coordinate and control efficient pilus assembly at the usher platform. As well as providing new information about the protein-protein interactions that determine pilus biogenesis, the results highlight the power of noncovalent MS to interrogate biological mechanisms, especially in complex mixtures of species. PMID:22371487

  7. The role of chaperone-subunit usher domain interactions in the mechanism of bacterial pilus biogenesis revealed by ESI-MS.

    Science.gov (United States)

    Morrissey, Bethny; Leney, Aneika C; Toste Rêgo, Ana; Phan, Gilles; Allen, William J; Verger, Denis; Waksman, Gabriel; Ashcroft, Alison E; Radford, Sheena E

    2012-07-01

    The PapC usher is a β-barrel outer membrane protein essential for assembly and secretion of P pili that are required for adhesion of pathogenic E. coli, which cause the development of pyelonephritis. Multiple protein subunits form the P pilus, the highly specific assembly of which is coordinated by the usher. Despite a wealth of structural knowledge, how the usher catalyzes subunit polymerization and orchestrates a correct and functional order of subunit assembly remain unclear. Here, the ability of the soluble N-terminal (UsherN), C-terminal (UsherC2), and Plug (UsherP) domains of the usher to bind different chaperone-subunit (PapDPapX) complexes is investigated using noncovalent electrospray ionization mass spectrometry. The results reveal that each usher domain is able to bind all six PapDPapX complexes, consistent with an active role of all three usher domains in pilus biogenesis. Using collision induced dissociation, combined with competition binding experiments and dissection of the adhesin subunit, PapG, into separate pilin and adhesin domains, the results reveal why PapG has a uniquely high affinity for the usher, which is consistent with this subunit always being displayed at the pilus tip. In addition, we show how the different soluble usher domains cooperate to coordinate and control efficient pilus assembly at the usher platform. As well as providing new information about the protein-protein interactions that determine pilus biogenesis, the results highlight the power of noncovalent MS to interrogate biological mechanisms, especially in complex mixtures of species.

  8. Highly conserved small subunit residues influence rubisco large subunit catalysis.

    Science.gov (United States)

    Genkov, Todor; Spreitzer, Robert J

    2009-10-30

    The chloroplast enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyzes the rate-limiting step of photosynthetic CO(2) fixation. With a deeper understanding of its structure-function relationships and competitive inhibition by O(2), it may be possible to engineer an increase in agricultural productivity and renewable energy. The chloroplast-encoded large subunits form the active site, but the nuclear-encoded small subunits can also influence catalytic efficiency and CO(2)/O(2) specificity. To further define the role of the small subunit in Rubisco function, the 10 most conserved residues in all small subunits were substituted with alanine by transformation of a Chlamydomonas reinhardtii mutant that lacks the small subunit gene family. All the mutant strains were able to grow photosynthetically, indicating that none of the residues is essential for function. Three of the substitutions have little or no effect (S16A, P19A, and E92A), one primarily affects holoenzyme stability (L18A), and the remainder affect catalysis with or without some level of associated structural instability (Y32A, E43A, W73A, L78A, P79A, and F81A). Y32A and E43A cause decreases in CO(2)/O(2) specificity. Based on the x-ray crystal structure of Chlamydomonas Rubisco, all but one (Glu-92) of the conserved residues are in contact with large subunits and cluster near the amino- or carboxyl-terminal ends of large subunit alpha-helix 8, which is a structural element of the alpha/beta-barrel active site. Small subunit residues Glu-43 and Trp-73 identify a possible structural connection between active site alpha-helix 8 and the highly variable small subunit loop between beta-strands A and B, which can also influence Rubisco CO(2)/O(2) specificity.

  9. Ancillary procedure for early diagnosis of brain damage in children

    International Nuclear Information System (INIS)

    Sumi, Masatoshi; Sha, Tenei; Ryo, Fukko; Kagawa, Kotaro.

    1979-01-01

    CT scan of the head was performed on 14 patients with cerebral palsy, 16 with central coordination disorders, and 16 controls, and findings showing cerebral atrophy and enlargement of the cerebral ventricle were obtained in cases both of cerebral palsy and of central coordination disorders. To objectify these findings, 10 items were selected and evaluated according to 4 grades (0 - 3) and were compared. As a result, it was concluded that CT scan is an excellent ancillary procedure for early diagnosis of brain damages. (Tsunoda, M.)

  10. 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

  11. 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.

  12. Energy and ancillary service dispatch through dynamic optimal power flow

    International Nuclear Information System (INIS)

    Costa, A.L.; Costa, A. Simoes

    2007-01-01

    This paper presents an approach based on dynamic optimal power flow (DOPF) to clear both energy and spinning reserve day-ahead markets. A competitive environment is assumed, where agents can offer active power for both demand supply and ancillary services. The DOPF jointly determines the optimal solutions for both energy dispatch and reserve allocation. A non-linear representation for the electrical network is employed, which is able to take transmission losses and power flow limits into account. An attractive feature of the proposed approach is that the final optimal solution will automatically meet physical constraints such as generating limits and ramp rate restrictions. In addition, the proposed framework allows the definition of multiple zones in the network for each time interval, in order to ensure a more adequate distribution of reserves throughout the power system. (author)

  13. Capabilities and costs for ancillary services provision by wind power plants. Deliverable D 3.1

    DEFF Research Database (Denmark)

    Faiella, Mariano; Hennig, Tobias; Cutululis, Nicolaos Antonio

    This report is the deliverable of the third work package of the REserviceS project and describes the technical options and related costs for the provision of ancillary services specifically from wind energy technologies. It is focused on the set of ancillary services defined in the previous work...

  14. Use of Ancillary Tests When Determining Brain Death in Pediatric Patients in the United States.

    Science.gov (United States)

    Lewis, Ariane; Adams, Nellie; Chopra, Arun; Kirschen, Matthew P

    2017-10-01

    Although pediatric brain death guidelines stipulate when ancillary testing should be used during brain death determination, little is known about the way these recommendations are implemented in clinical practice. We conducted a survey of pediatric intensivists and neurologists in the United States on the use of ancillary testing. Although most respondents noted they only performed an ancillary test if the clinical examination and apnea test could not be completed, 20% of 195 respondents performed an ancillary test for other reasons, including (1) to convince a family that objected to the brain death determination that a patient is truly dead (n = 21), (2) personal preference (n = 14), and (3) institutional requirement (n = 5). Our findings suggest that pediatricians use ancillary tests for a variety of reasons during brain death determination. Medical societies and governmental regulatory bodies must reinforce the need for homogeneity in practice.

  15. 75 FR 14342 - Market-Based Rates for Wholesale Sales of Electric Energy, Capacity and Ancillary Services by...

    Science.gov (United States)

    2010-03-25

    ...; Order No. 697-D] Market-Based Rates for Wholesale Sales of Electric Energy, Capacity and Ancillary... affiliates.\\3\\ \\1\\ Market-Based Rates for Wholesale Sales of Electric Energy, Capacity and Ancillary Services...\\ Market-Based Rates for Wholesale Sales of Electric Energy, Capacity and Ancillary Services by Public...

  16. 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...

  17. 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

  18. Estimating Net Primary Productivity Using Satellite and Ancillary Data

    Science.gov (United States)

    Choudhury, Bhaskar J.

    2002-01-01

    The net primary productivity (C) or the annual rate of carbon accumulation per unit ground area by terrestrial plant communities is the difference of gross photosynthesis (A(sub g)) and respiration (R) per unit ground area. Available field observations show that R is a large and variable fraction of A(sub g), although it is generally recognized that there are considerable difficulties in determining these fluxes, and thus pose challenge in assessing the accuracy. Further uncertainties arise in extrapolating field measurements (which are acquired over a hectare or so area) to regional scale. Here, an approach is presented for determining these fluxes using satellite and ancillary data to be representative of regional scale and allow assessment of interannual variation. A, has been expressed as the product of radiation use efficiency for gross photosynthesis by an unstressed canopy and intercepted photosynthetically active radiation, which is then adjusted for stresses due to soil water shortage and temperature away from optimum. R has been calculated as the sum of growth and maintenance components (respectively, R(sub g) and R(sub m)).The R(sub m) has been determined from nitrogen content of plant tissue per unit ground area, while R(sub g) has been obtained as a fraction of the difference of A(sub g) and R(sub m). Results for five consecutive years (1986-1990) are presented for the Amazon-Tocontins, Mississippi, and Ob River basins.

  19. 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.

  20. 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...

  1. Characterization of fimbrial subunits from Bordetella species

    NARCIS (Netherlands)

    Mooi, F.R.; Heide, H.G.J. van der; Avest, A.R. ter; Welinder, K.G.; Livey, I.; Zeijst, B.A.M. van der; Gaastra, W.

    Using antisera raised against serotype 2 and 3 fimbrial subunits from Bordetella pertussis, serologically related polypeptides were detected in Bordetella bronchiseptica, Bordetella parapertussis and Bordetella avium strains. The two B. pertussis fimbrial subunits, and three of the serologically

  2. 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...

  3. 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...

  4. 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 &...

  5. Technical and economic aspects of ancillary services markets in the electric power industry: an international comparison

    International Nuclear Information System (INIS)

    Raineri, R.; Rios, S.; Schiele, D.

    2006-01-01

    We present a comparative analysis of technical and economic aspects of ancillary services on the markets of England and Wales, Nordic Countries, California, Argentina, Australia and Spain, comparing the services of voltage control, frequency regulation and system restoration. All the analyzed markets rely on the existence of an administrator of ancillary services, function that leads to the figure of the system operator. Among the services analyzed, the mandatory nature of voltage control and primary frequency regulation stands out, being both the ancillary services with the higher market price and the shortest period of time requirements. In general, the recognized costs of the services correspond to investments operation, maintenance, and opportunity costs. In the provision of these ancillary services, there are no clear preferences for a particular resource allocation mechanism, where mandatory provision, auctions, competitive offers and different time length bilateral contracts are combined

  6. 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...

  7. 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...

  8. 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...

  9. 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...

  10. 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:...

  11. 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 &...

  12. 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...

  13. 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...

  14. 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...

  15. 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...

  16. 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 &...

  17. 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:...

  18. 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 &...

  19. 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...

  20. 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...

  1. 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:...

  2. 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...

  3. 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...

  4. 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...

  5. 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...

  6. 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...

  7. 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...

  8. 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...

  9. 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...

  10. 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:...

  11. 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...

  12. 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...

  13. 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 &...

  14. EX1504L2 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 EX1504L2: Campaign to Address...

  15. 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...

  16. 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...

  17. 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...

  18. 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...

  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. 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...

  1. 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...

  2. 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...

  3. 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...

  4. 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 &...

  5. 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 &...

  6. 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...

  7. 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...

  8. 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...

  9. 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 &...

  10. 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...

  11. 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:...

  12. The Nordic approach to market-based provision of ancillary services

    International Nuclear Information System (INIS)

    Kristiansen, Tarjei

    2007-01-01

    This paper presents an overview of the markets for ancillary services, which are regulated and traded differently within and across borders in the Nordic countries. We describe the services provided and their characteristics in terms of definition, participation, contracting approach and duration, selection of offers, and dispatch criteria. Further, we assess the costs, specify the payments, and discuss cost allocation, and we conclude with a policy discussion of ancillary services in the Nordic countries. (author)

  13. 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.

  14. Estimating Next Primary Productivity using Satellite and Ancillary Data

    Science.gov (United States)

    Choudhury, B. J.

    The net primary productivity (C) or annual rate of carbon accumulation per unit ground area by terrestrial plant communities is the difference of the rate of gross photosynthesis (Ag) and autotrophic respiration (R) per unit ground area. Although available observations show that R is a large and variable fraction of Ag, viz., 0.3 to 0.7, it is generally recognized that much uncertainties exist in this fraction due to difficulties associated with the needed measurements. Additional uncertainties arise when these measurements are extrapolated to regional or global land surface using empirical equations, for example, using regression equations relating C to mean annual precipitation and air temperature. Here, a process- based approach has been taken to calculate Ag and R using satellite and ancillary data. Ag has been expressed as a product of radiation use efficiency, magnitude of intercepted photosynthetically active radiation (PAR), and normalized by stresses due to soil water shortage and air temperature away from the optimum range. A biophysical model has been used to determine the radiation use efficiency from the maximum rate of carbon assimilation by a leaf, foliage temperature, and the fraction of diffuse PAR incident on a canopy. All meteorological data (PAR, air temperature, precipitation, etc.) needed for the calculation are derived from satellite observations, while a land use, land cover data (based on satellite and ground measurements) have been used to assess the maximum rate of carbon assimilation by a leaf of varied cover type based on field measurements. R has been calculated as the sum of maintenance and growth components. The maintenance respiration of foliage and live fine roots at a standard temperature of different land cover has been determined from their nitrogen content using field and satellite measurements, while that of living fraction of woody stem (viz., sapwood) from the seasonal maximum leaf area index as determined from satellite

  15. Subunit Stoichiometry of Human Muscle Chloride Channels

    OpenAIRE

    Fahlke, Christoph; Knittle, Timothy; Gurnett, Christina A.; Campbell, Kevin P.; George, Alfred L.

    1997-01-01

    Voltage-gated Cl? channels belonging to the ClC family appear to function as homomultimers, but the number of subunits needed to form a functional channel is controversial. To determine subunit stoichiometry, we constructed dimeric human skeletal muscle Cl? channels in which one subunit was tagged by a mutation (D136G) that causes profound changes in voltage-dependent gating. Sucrose-density gradient centrifugation experiments indicate that both monomeric and dimeric hClC-1 channels in their ...

  16. Colombian ancillary services and international connections: Current weaknesses and policy challenges

    International Nuclear Information System (INIS)

    Carvajal, S.X.; Serrano, J.; Arango, S.

    2013-01-01

    Ancillary services are required to maintain the unity, stability, and quality of power systems. In Colombia these services are required to operate the national power system and the international connections with the neighboring countries. The system is influenced by factors ranging from system's topology to social and political aspects, such as the large number of terrorist attacks. In light of these particularities, we consider Colombia as a learning lab for ancillary services in the region. Colombia's power system relies on three ancillary services for its operation, namely frequency regulation, voltage control, and blackstart service. From 2010 Special Protection Systems were also added. In this paper we first analyze the technical aspects, operational restrictions, financial management, and the most relevant regulatory conditions of these ancillary services of the SIN. We also take into consideration the main regulatory characteristics and statistical data related to energy exchanges that have taken place between Colombia and Venezuela and Ecuador. Thereafter, we depict the main weaknesses and policy challenges that Colombia must address in order to increase the effectiveness and coverage of ancillary services in both the SIN and in the international interconnections. Finally, we propose new market oriented regulations to encourage investments and new tools for international connections. - Highlights: ► Blackout of 2007 forces Colombian regulator to improve security of power system. ► Four ancillary services are currently used for secure operation of Colombian system. ► Special Protection Systems service added in 2010. ► Frequency control is the only remunerated service in Colombia. ► Legislation should exist for remuneration of voltage control and black start services.

  17. 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

  18. 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.

  19. Coordinated Control Scheme for Ancillary Services from Offshore Wind Power Plants to AC and DC Grids

    DEFF Research Database (Denmark)

    Sakamuri, Jayachandra N.; Altin, Müfit; Hansen, Anca Daniela

    2016-01-01

    This paper proposes a new approach of providing ancillary services to AC and DC grids from offshore wind power plants (OWPPs), connected through multi-terminal HVDC network. A coordinated control scheme where OWPP’s AC grid frequency modulated according to DC grid voltage variations is used...... to detect and provide the ancillary service requirements of both AC and DC grids, is proposed in this paper. In particular, control strategies for onshore frequency control, fault ridethrough support in the onshore grid, and DC grid voltage control are considered. The proposed control scheme involves only...

  20. 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.

  1. 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....

  2. 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...

  3. 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...

  4. 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...

  5. 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...

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

    African Journals Online (AJOL)

    AJRH Managing Editor

    income countries where HIV incidence is high, but the benefits of research are ... provision of ancillary care – medical care provided to clinical trial participants during a ... to or use of quality delivery services is an issue of ... with drastically reduced costs, questions arise over .... The 'partial entrustment' model differentiates.

  7. 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

    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...

  8. Evaluation of three ancillary treatments in the management of equine grass sickness.

    Science.gov (United States)

    Fintl, C; McGorum, B C

    2002-09-28

    Brotizolam, acetylcysteine and aloe vera gel were evaluated as ancillary treatments for 29 cases of equine grass sickness. None of the treatments had any significant beneficial effect on the survival of the horses. However, 11 of 13 horses with mild chronic grass sickness survived solely with intensive nursing care.

  9. Management of Ovarian Cysts by Laparoscopic Extracorporeal Approach Using Single Ancillary Trocar

    Directory of Open Access Journals (Sweden)

    Metin Capar

    2009-12-01

    Conclusion: This technique does not require the use of two or more ancillary trocars or widening of the trocar incision. The duration of surgery can be shortened considerably and complete excision of the cystic capsule can be performed. Homeostasis was achieved using 3-0 polyglactin sutures. No electrocoagulation was required.

  10. Island operation capability in the Colombian electrical market: a promising ancillary service of distributed energy resources

    DEFF Research Database (Denmark)

    Marín-Jiménez, Juan D.; Carvajal-Quintero, Sandra; Guerrero, Josep M.

    2018-01-01

    is presented. Subsequently, an existing electrical distribution system with small hydropower plants is considered as a study case in order to identify the technical requirements set out for both, the distribution system and the generation source. Finally, taking into account the current Law and regulations......, a proposal is outlined for the implementation of the island operation capability ancillary service in the Colombian electrical system....

  11. 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...

  12. 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.

  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. 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

  15. 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.

  16. 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.

  17. 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

  18. Provision of enhanced ancillary services from wind power plants - Examples and challenges

    DEFF Research Database (Denmark)

    Hansen, Anca Daniela; Altin, Müfit; Iov, Florin

    2016-01-01

    Emphasis in this article is on the power system impact of wind power plants capability to provide enhanced ancillary services, i.e. temporary frequency response (TFR) and power oscillation damping (POD). The main objective of the article is to analyze and justify the challenges in the use of TFR...... and POD from wind power plants (WPPs). The study is conducted with an aggregated wind power plant model which is integrated into a generic power system model, specifically designed to assess the targeted ancillary services in a relatively simple, but still relevant environment. Various case studies...... with different wind power penetration levels are considered. The study shows that WPPs can provide additional control features such as TFR and POD to enhance the stability of power systems with large share of wind power. Nevertheless, the results illustrate that the power system stability can be potentially...

  19. 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.

  20. 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

  1. 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

    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....... 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...

  2. Simulated annealing to handle energy and ancillary services joint management considering electric vehicles

    DEFF Research Database (Denmark)

    Sousa, Tiago M; Soares, Tiago; Morais, Hugo

    2016-01-01

    The massive use of distributed generation and electric vehicles will lead to a more complex management of the power system, requiring new approaches to be used in the optimal resource scheduling field. Electric vehicles with vehicle-to-grid capability can be useful for the aggregator players...... in the mitigation of renewable sources intermittency and in the ancillary services procurement. In this paper, an energy and ancillary services joint management model is proposed. A simulated annealing approach is used to solve the joint management for the following day, considering the minimization...... of the aggregator total operation costs. The case study considers a distribution network with 33-bus, 66 distributed generation and 2000 electric vehicles. The proposed simulated annealing is matched with a deterministic approach allowing an effective and efficient comparison. The simulated annealing presents...

  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. 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...

  5. Exploring the ancillary benefits of the Kyoto Protocol for air pollution in Europe

    International Nuclear Information System (INIS)

    Vuuren, D.P. van; Cofala, J.; Eerens, H.E.; Oostenrijk, R.; Heyes, C.; Klimont, Z.; Elzen, M.G.J. den; Amann, M.

    2006-01-01

    An integrated approach to climate change and regional air pollution can harvest considerable ancillary benefits in terms of environmental impacts and costs. This is because both problems are caused to a large extent by the same activity (fossil fuel combustion). Substantial ancillary benefits were found for regional air pollution (SO 2 , NO x , VOC and particulate matter) of implementing the Kyoto Protocol (intended to control greenhouse gas emissions) in Europe. For instance, while three different scenarios on Kyoto implementation were found to reduce European CO 2 emissions by 4-7%, they also reduced European emissions of SO 2 by 5-14% compared with a no Kyoto policies case. The magnitude of ancillary benefits depends on how flexible mechanisms and surplus emission allowances are used in meeting the Kyoto targets. The total cost savings for implementing current policies for regional air pollution of the Kyoto Protocol are of an order of 2.5-7 billion Euro. In all cases, this is in the order of half the costs of the climate policy (4-12 billion Euro). Using flexible mechanisms reduces emissions of air pollutants for Europe as a whole even further than domestic implementation (e.g. 10-14% versus 5% for SO 2 emissions), but the reductions are shifted from Western Europe to Central and Eastern Europe and Russia. The use of surplus emission allowances to achieve the Kyoto targets decreases the ancillary benefits, in particular for the latter group of countries (e.g. unprotected area against acidification increases from 1.3 to 1.7 million ha)

  6. Prevalence of influenza vaccination among nurses and ancillary workers in Italy: systematic review and meta analysis.

    Science.gov (United States)

    La Torre, Giuseppe; Mannocci, Alice; Ursillo, Paolo; Bontempi, Claudio; Firenze, Alberto; Panico, Maria Grazia; Sferrazza, Antonella; Ronga, Chiara; D'Anna, Adele; Amodio, Emanuele; Romano, Nino; Boccia, Antonio

    2011-07-01

    Italian Ministry of Health, recommends vaccination for seasonal influenza to all healthcare workers (HCW), particularly to nurses who have an important interaction with patients. The aim of this study is to conduct a systematic review in order to estimate the pooled prevalence of influenza vaccinations among nurses and ancillary workers in Italy and analyse the enhancing and hindering factors. The review was performed using 15 articles, six containing the prevalence of vaccination for nurses and ancillary workers, while the others qualitative analysis. In all the selected articles the score calculation has been carried out by using a protocol for observational studies. The nurses and ancillary workers pooled proportion of influenza vaccination was respectively 13.47% (95%CI 9.58-17.90%) and 12.52% (95%CI 9.97-15.31%). The Italian mean of influenza vaccination prevalence appear low if compared to other European countries, ranging from 15% to 29% in Countries such as UK, Germany, France. This situation of weakness should be seen as an opportunity to improve the vaccination rate for seasonal influenza significantly This should be done by intervening on the category which affirms caring less. In fact, this category has a priority to receive vaccination, due to their numbers and closer contact to patients. Research was conducted using medical database Scopus, PubMed, the search engine Google Scholar and ISI web of knowledge, and was concluded February 1st 2011.

  7. The impact of communication barriers on diagnostic confidence and ancillary testing in the emergency department.

    Science.gov (United States)

    Garra, Gregory; Albino, Hiram; Chapman, Heather; Singer, Adam J; Thode, Henry C

    2010-06-01

    Communication barriers (CBs) compromise the diagnostic power of the medical interview and may result in increased reliance on diagnostic tests or incorrect test ordering. The prevalence and degree to which these barriers affect diagnosis, testing, and treatment are unknown. To quantify and characterize CBs encountered in the Emergency Department (ED), and assess the effect of CBs on initial diagnosis and perceived reliance on ancillary testing. This was a prospective survey completed by emergency physicians after initial adult patient encounters. CB severity, diagnostic confidence, and reliance on ancillary testing were quantified on a 100-mm Visual Analog Scale (VAS) from least (0) to most (100). Data were collected on 417 ED patient encounters. CBs were reported in 46%; with a mean severity of 50 mm on a 100-mm VAS with endpoints of "perfect communication and "no communication." Language was the most commonly reported form of CB (28%). More than one CB was identified in 6%. The 100-mm VAS rating of diagnostic confidence was lower in patients with perceived CBs (64 mm) vs. those without CBs (80 mm), p Communication barriers in our ED setting were common, and resulted in lower diagnostic confidence and increased perception that ancillary tests are needed to narrow the diagnosis. Copyright 2010 Elsevier Inc. All rights reserved.

  8. 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.

  9. 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.

  10. 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.

  11. 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

    The impact of a high penetration of wind power generation in power systems motivates need for an assessment of its inte raction with electricity markets. With the continuous evolution of wind turbines technology, wind farms have today the ability to provide certain ancillary services...... with appropriate levels of security and reliability . The participation of wind f arms i n ancillary service market s ought to rely on market designs and offering strategies that satisfy power system needs, as well as their operating characteristics . Here we evaluate the system impact of different offering...... 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...

  12. 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)

  13. Ancillary Services 4.0: A Top-To-Bottom Control-Based Approach for Solving Ancillary Services Problems in Smart Grids

    DEFF Research Database (Denmark)

    De Zotti, Giulia; Pourmousavi, S. Ali; Madsen, Henrik

    2018-01-01

    of AS are not able to benefit from the flexible energy resources. They also cannot cope with the new level of stochasticity, non-linearity, and dynamics of generation and flexibility. To overcome such issues and exploit the potential of flexibility resources, a new strategy is required. In this paper......Power systems are experiencing a large amount of renewable generation with highly stochastic and partly unpredictable characteristics. This change in energy production implies significant consequences related to the provision of ancillary services (AS). Current markets dedicated to the provision......, by capitalizing on flexibility resources' potential, AS 4.0 approach is proposed, which offers a comprehensive solution for the AS provision in the smart grid era....

  14. 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

  15. 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

  16. Resource and revenue potential of California residential load participation in ancillary services

    International Nuclear Information System (INIS)

    Mathieu, Johanna L.; Dyson, Mark E.H.; Callaway, Duncan S.

    2015-01-01

    Increasing penetrations of intermittent renewable energy resources will require additional power system services. California recently adopted an energy storage mandate to support its renewable portfolio standard, which requires 33% of delivered energy from renewables by 2020. The objective of this paper is to estimate the amount of energy storage that could be provided by residential thermostatically controlled loads, such as refrigerators and air conditioners, and the amount of revenue that could be earned by loads participating in ancillary services markets. We model load aggregations as virtual energy storage, and use simple dynamical system models and publicly available data to generate our resource and revenue estimates. We find that the resource potential is large: 10–40 GW/8–12 GWh, which is significantly more than that required by the mandate. We also find that regulation and spinning/non-spinning reserve revenues vary significantly depending upon type of load and, for heat pumps and air conditioners, climate zone. For example, mean regulation revenues for refrigerators are $11/year, for electric water heaters are $24/year, for air conditioners are $0-32/year, and for heat pumps are $22–56/year. Both consumer choices, such as appliance settings, and policy, such as the design of ancillary service compensation and appliance standards, could increase revenue potentials. - Highlights: • California's energy storage mandate requires 1.325 GW of energy storage by 2020. • Residential loads such as refrigerators have thermal energy storage. • California's residential loads could provide 10-40 GW/8-12 GWh of storage. • Loads participating in ancillary services markets could earn up to $56/load/year. • Consumer choices and policy mechanisms could increase revenue potentials

  17. Evaluation of Contribution for Voltage Control Ancillary Services Based on Social Surplus

    Science.gov (United States)

    Ueki, Yuji; Hara, Ryoichi; Kita, Hiroyuki; Hasegawa, Jun

    Reactive power supply plays an important role in active power supply with adequate system voltages. Various pricing mechanism for reactive power supply have been developed and some of them are adopted in some power systems, however they are in a trial stage. The authors also focus on development of a pricing method for reactive power ancillary services. This problem involves two technical issues: rational estimation of the cost associated with reactive power supply and fair and transparent allocation of the estimated cost among the market participants. This paper proposes methods for evaluating the contribution of generators and demands.

  18. Opportunities for ice storage to provide ancillary services to power grids incorporating wind turbine generation

    Science.gov (United States)

    Finley, Christopher

    Power generation using wind turbines increases the electrical system balancing, regulation and ramp rate requirements due to the minute to minute variability in wind speed and the difficulty in accurately forecasting wind speeds. The addition of thermal energy storage, such as ice storage, to a building's space cooling equipment increases the operational flexibility of the equipment by allowing the owner to choose when the chiller is run. The ability of the building owner to increase the power demand from the chiller (e.g. make ice) or to decrease the power demand (e.g. melt ice) to provide electrical system ancillary services was evaluated.

  19. 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.

  20. 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

    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...... 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...

  1. 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

  2. Crisis in our hospital kitchens: ancillary staffing levels during an outbreak of food poisoning in a long stay hospital.

    Science.gov (United States)

    Pollock, A M; Whitty, P M

    1990-02-10

    An investigation into an outbreak of food poisoning caused by Clostridium perfringens showed evidence of poor food handling by catering staff. The reasons behind this were explored by interviewing catering staff, analysing shifts and rotas, and looking at staff vacancies. Morale was low because of staff shortages resulting from a long term recruitment problem. In consequence staff were working double shifts and often for weeks on end without a day off. The reasons for the recruitment problem included the difficulty of recruiting semiskilled labour from a middle class area, low wages, lack of management support, and the poor image of the hospital as a place of work. Similar factors affect the recruitment and retention of ancillary staff nationally. The NHS has a poor record as an employer of ancillary staff, paying lower wages than other organisations for equivalent posts. Competitive tendering has further worsened the position of ancillary staff, with the result that good quality of care and service has often not been achieved. The NHS Review, with its emphasis on quality of care, makes no mention of ancillary staff. Yet high standards of ancillary provision are essential if further outbreaks of food poisoning in hospitals are to be prevented.

  3. 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.

  4. MR imaging diagnosis of posterior cruciate ligament injury: importance of ancillary findings

    International Nuclear Information System (INIS)

    Hwang, Kang Ik; Lee, Jong Hwa; Kim, Young Sun; Lee, Jung Hwoi; Ki, Tae Sung; Park, Jong Oag

    1997-01-01

    To evaluate the importance of two ancillary findings of anterior tibial plateau bruise/fracture and popliteus muscle strain on MR diagnosis of posterior cruicate ligament injury. We retrospectively evaluated 48 patients with confirmed posterior cruciate ligament tear. We studied the incidence of anterior tibial plateau injury and popliteus muscle strain, and the specificity of popliteus muscle strain with or without bony injury. A complete tear of the posterior cruciate ligament was noted in 37 cases, a partial tear in 11. Anterior tibial plateau lesion was found in 21 of 48 cases (44%); This total was made up of 17/37 PCL complete tears (46%) and 4/11 partial tears (36%). The difference in the incidence of complete and partial tears is not statistically significant. Popliteus muscle injury was found in 20 of 48 cases (42%), the total consisted of 19/37 PCL complete tears (51%) and 1/11 partial tears (10%). The incidence of 42% is relatively high, approximating that of bony injury. The difference in the incidence of complete and partial tears is statistically significant (p < 0.006). Specificity for posterior cruciate ligament tear is 69%(20/29), and when concomitant with anterior tibial plateau injury is 94%(16/17). As in the case of anterior cruciate ligament injury, these documented ancillary findings of anterior tibial plateau and popliteus muscle injuries are very helpful when MR diagnosis of posterior cruciate ligament injury itself and differentiation of partial and complete rupture are doubtful

  5. 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.

  6. 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.

  7. An integrated pan-European ancillary services market for frequency control

    International Nuclear Information System (INIS)

    Scherer, Marc; Zima, Marek; Andersson, Göran

    2013-01-01

    Real-time balancing of mismatches between consumption and production is one of the key elements for the secure operation of power systems. This takes place within the framework of ancillary services managed by respective national transmission system operating companies. The objective of this paper is to investigate the option of one integrated pan-European ancillary services market. Our contribution is twofold: quantifying the potential benefit of such an option and outlining a possible approach to such an option highlighting its positive properties as well as risks and challenges. In several recently published considerations of pan-European electricity supply models this topic has not been adequately addressed, but we believe it is one of the crucial subjects in shaping the future electricity supply in Europe. -- Author-Highlights: •We investigated the option of a one-area market for frequency control in Continental Europe. •For the current situation a centralised approach may lead to a significant reduction of control reserves. •In the long run it is a possibility to avoid a tremendous increase of needed control reserves. •The implementation of an integrated market is not to be underestimated

  8. Pipe stress analysis on HCCR-TBS ancillary systems in conceptual design

    Energy Technology Data Exchange (ETDEWEB)

    Ahn, Mu-Young, E-mail: myahn74@nfri.re.kr [National Fusion Research Institute, Daejeon (Korea, Republic of); Cho, Seungyon [National Fusion Research Institute, Daejeon (Korea, Republic of); Lee, Eo Hwak [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of); Park, Yi-Hyun; Lee, Youngmin [National Fusion Research Institute, Daejeon (Korea, Republic of)

    2016-11-01

    Highlights: • Pipe stress is performed on Korean HCCR-TBS for the load combinations including seismic events. • The resultant stress meets the requirement of the design code & standard except one position where modification is needed. • The results gives useful information for the design evolution in the next desgin phase. - Abstract: Korean Helium Cooled Ceramic Reflector (HCCR) Test Blanket System (TBS) will be tested in ITER to demonstrate feasibility of the breeding blanket concept. The HCCR-TBS comprises Test Blanket Module (TBM) with associated shield, and ancillary systems located in various positions of ITER building. Currently, conceptual design for the HCCR-TBS is in progress. This paper presents pipe stress analysis results for the HCCR-TBS ancillary systems. The pipe stress analysis was performed in accordance with ASME B31.3 for major pipes of the Helium Cooling System (HCS) and the Coolant Purification System (CPS), which are operated in high pressure and temperature. The pipe stress for various load cases and load combinations were calculated. Operational pressure and temperature during plasma operation are applied as pressure load and thermal load, respectively. In addition seismic events were combined to investigate the code compliance for sustained load case and occasional load case. It was confirmed that the resultant stress meets the requirements of ASME B31.3 except one position in which it needs modification. These results give useful information for the next design phase, for example, nozzle loads for the component selection, the support design parameters, etc.

  9. Optimal bidding strategy for microgrids in joint energy and ancillary service markets considering flexible ramping products

    International Nuclear Information System (INIS)

    Wang, Jianxiao; Zhong, Haiwang; Tang, Wenyuan; Rajagopal, Ram; Xia, Qing; Kang, Chongqing; Wang, Yi

    2017-01-01

    Highlights: •Flexible ramping products are modelled in the framework of a microgrid. •Microgrids’ optimal bidding model is proposed in energy and ancillary service markets. •A hybrid stochastic and robust optimization approach is adopted. •The effectiveness of the proposed bidding model is verified based on real-world data. -- Abstract: Due to the volatile nature of wind and photovoltaic power, wind farms and solar stations are generally thought of as the consumers of ramping services. However, a microgrid (MG) is able to strategically integrate various distributed energy resources (DERs) to provide both energy and ancillary services (ASs) for the bulk power system. To evaluate the ramping capabilities of an MG in the joint energy and AS markets, an optimal bidding strategy is developed in this paper considering flexible ramping products (FRPs). By aggregating and coordinating various DERs, including wind turbines (WTs), photovoltaic systems (PVs), micro-turbines (MTs) and energy storage systems (ESSs), the MG is able to optimally allocate the capacities for energy, spinning reserve and ramping. Taking advantage of the synergy among DERs, the MG can maximize its revenues from different markets. Moreover, the flexibility of the MG for the bulk power system can be fully explored. To address the uncertainties introduced by renewable generation and market prices, a hybrid stochastic/robust optimization (RO) approach is adopted. Case studies based on a real-world MG with various DERs demonstrate the market behavior of the MG using the proposed bidding model.

  10. Pipe stress analysis on HCCR-TBS ancillary systems in conceptual design

    International Nuclear Information System (INIS)

    Ahn, Mu-Young; Cho, Seungyon; Lee, Eo Hwak; Park, Yi-Hyun; Lee, Youngmin

    2016-01-01

    Highlights: • Pipe stress is performed on Korean HCCR-TBS for the load combinations including seismic events. • The resultant stress meets the requirement of the design code & standard except one position where modification is needed. • The results gives useful information for the design evolution in the next desgin phase. - Abstract: Korean Helium Cooled Ceramic Reflector (HCCR) Test Blanket System (TBS) will be tested in ITER to demonstrate feasibility of the breeding blanket concept. The HCCR-TBS comprises Test Blanket Module (TBM) with associated shield, and ancillary systems located in various positions of ITER building. Currently, conceptual design for the HCCR-TBS is in progress. This paper presents pipe stress analysis results for the HCCR-TBS ancillary systems. The pipe stress analysis was performed in accordance with ASME B31.3 for major pipes of the Helium Cooling System (HCS) and the Coolant Purification System (CPS), which are operated in high pressure and temperature. The pipe stress for various load cases and load combinations were calculated. Operational pressure and temperature during plasma operation are applied as pressure load and thermal load, respectively. In addition seismic events were combined to investigate the code compliance for sustained load case and occasional load case. It was confirmed that the resultant stress meets the requirements of ASME B31.3 except one position in which it needs modification. These results give useful information for the next design phase, for example, nozzle loads for the component selection, the support design parameters, etc.

  11. A hybrid PSO technique for procuring VAR ancillary service in the deregulated electricity markets

    Energy Technology Data Exchange (ETDEWEB)

    El-Araby, E.E. [Department of Electrical Engineering, Suez Canal University (Egypt); Yorino, Naoto [Department of Artificial Complex Systems Engineering, Hiroshima University (Japan)

    2010-07-15

    This paper develops a new market-based technique for acquiring VAR ancillary service in the electricity market. The main objective of the developed market is to enable transmission operator ''TO'' to procure VAR service in a long term contract from the critical VAR providers that satisfy minimum VAR service payment while maintaining system security. Reactive power control problem for voltage stability is introduced into the VAR market problem in an explicit manner for normal and emergency states. An integration of particle swarm optimization ''PSO'' is presented with successive linear programming ''SLP'' for dealing with the VAR ancillary service problem. The problem is formulated as a large-scale nonlinear constrained optimization problem with a non-differentiable objective function representing VAR payment and operational costs. This type of problem is hard to be treated straightforwardly by the classical optimization methods. Therefore, we propose here a two-layer hybrid PSO/SLP approach, which is suited for carrying out the difficulties associated with non-differentiable and discontinuous objective functions. The proposed method has been examined on the standard IEEE 57 bus-system and compared with GA/SLP method to demonstrate its capability. (author)

  12. Muscular subunits transplantation for facial reanimation

    Directory of Open Access Journals (Sweden)

    Hazan André Salo Buslik

    2006-01-01

    Full Text Available PURPOSE: To present an alternative technique for reconstruction of musculocutaneous damages in the face transferring innervated subsegments(subunits of the latissimus dorsi flap for replacement of various facial mimetic muscles. METHODS: One clinical case of trauma with skin and mimetic muscles damage is described as an example of the technique. The treatment was performed with microsurgical transfer of latissimus dorsi muscle subunits. Each subunit present shape and dimensions of the respective mimetic muscles replaced. The origin, insertions and force vectors for the mimicmuscle lost were considered. Each subsegment has its own arterial and venous supply with a motor nerve component for the muscular unit. RESULTS: Pre and one year postoperative photos registration of static and dynamic mimic aspects, as well as digital electromyography digital data of the patients were compared. The transplanted muscular units presented myoeletric activity, fulfilling both the functional and cosmetic aspect. CONCLUSION: This technique seems to be a promising way to deal with the complex musculocutaneous losses of the face as well as facial palsy.

  13. Influvac, a trivalent inactivated subunit influenza vaccine.

    Science.gov (United States)

    Zuccotti, Gian Vincenzo; Fabiano, Valentina

    2011-01-01

    Influenza represents a major sanitary and socio-economic burden and vaccination is universally considered the most effective strategy for preventing the disease and its complications. Traditional influenza vaccines have been on the market since the late 1940s, with million of doses administered annually worldwide, and demonstrated a substantial efficacy and safety. The trivalent inactivated subunit vaccine has been available for more than 25 years and has been studied in healthy children, adults and the elderly and in people affected by underlying chronic medical conditions. We describe vaccine technology focusing on subunit vaccine production procedures and mode of action and provide updated information on efficacy and safety available data. A review of efficacy and safety data in healthy subjects and in high risk populations from major sponsor- and investigator-driven studies. The vaccine showed a good immunogenicity and a favorable safety profile in all target groups. In the panorama of actually available influenza vaccines, trivalent inactivated subunit vaccine represents a well-established tool for preventing flu and the associated complications.

  14. 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

  15. Soybean glycinin subunits: Characterization of physicochemical and adhesion properties.

    Science.gov (United States)

    Mo, Xiaoqun; Zhong, Zhikai; Wang, Donghai; Sun, Xiuzhi

    2006-10-04

    Soybean proteins have shown great potential for applications as renewable and environmentally friendly adhesives. The objective of this work was to study physicochemical and adhesion properties of soy glycinin subunits. Soybean glycinin was extracted from soybean flour and then fractionated into acidic and basic subunits with an estimated purity of 90 and 85%, respectively. Amino acid composition of glycinin subunits was determined. The high hydrophobic amino acid content is a major contributor to the solubility behavior and water resistance of the basic subunits. Acidic subunits and glycinin had similar solubility profiles, showing more than 80% solubility at pH 2.0-4.0 or 6.5-12.0, whereas basic subunits had considerably lower solubility with the minimum at pH 4.5-8.0. Thermal analysis using a differential scanning calorimeter suggested that basic subunits form new oligomeric structures with higher thermal stability than glycinin but no highly ordered structures present in isolated acidic subunits. The wet strength of basic subunits was 160% more than that of acidic subunits prepared at their respective isoelectric points (pI) and cured at 130 degrees C. Both pH and the curing temperature significantly affected adhesive performance. High-adhesion water resistance was usually observed for adhesives from protein prepared at their pI values and cured at elevated temperatures. Basic subunits are responsible for the water resistance of glycinin and are a good starting material for the development of water-resistant adhesives.

  16. 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

  17. Disassembly of JT-60 tokamak device and ancillary facilities for JT-60 tokamak

    International Nuclear Information System (INIS)

    Okano, Fuminori; Ichige, Hisashi; Miyo, Yasuhiko; Kaminaga, Atsushi; Sasajima, Tadayuki; Nishiyama, Tomokazu; Yagyu, Jun-ichi; Ishige, Youichi; Suzuki, Hiroaki; Komuro, Kenichi; Sakasai, Akira; Ikeda, Yoshitaka

    2014-03-01

    The disassembly of JT-60 tokamak device and its peripheral equipments, where the total weight was about 5400 tons, started in 2009 and accomplished in October 2012. This disassembly was required process for JT-60SA project, which is the Satellite Tokamak project under Japan-EU international corroboration to modify the JT-60 to the superconducting tokamak. This work was the first experience of disassembling a large radioactive fusion device based on Radiation Hazard Prevention Act in Japan. The cutting was one of the main problems in this disassembly, such as to cut the welded parts together with toroidal field coils, and to cut the vacuum vessel into two. After solving these problems, the disassembly completed without disaster and accident. This report presents the outline of the JT-60 disassembly, especially tokamak device and ancillary facilities for tokamak device. (author)

  18. 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.

  19. Estimating commercial property prices: an application of cokriging with housing prices as ancillary information

    Science.gov (United States)

    Montero-Lorenzo, José-María; Larraz-Iribas, Beatriz; Páez, Antonio

    2009-12-01

    A vast majority of the recent literature on spatial hedonic analysis has been concerned with residential property values, with only very few examples of studies focused on commercial property prices. The dearth of studies can be attributed to some of the challenges faced in the analysis of commercial properties, in particular the scarcity of information compared to residential transactions. In order to address this issue, in this paper we propose the use of cokriging and housing prices as ancillary information to estimate commercial property prices. Cokriging takes into account the spatial autocorrelation structure of property prices, and the use of more abundant information on housing prices helps to improve the accuracy of property value estimates. A case study of Toledo in Spain, a city for which commercial activity stemming from tourism is one of the key elements of the economy in the city, demonstrates that substantial accuracy and precision gains can be obtained from the use of cokriging.

  20. 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.

  1. Ancillary services and the integration of substantial quantities of wind power

    DEFF Research Database (Denmark)

    Østergaard, Poul Alberg

    2006-01-01

    . Ballerup: Elkraft System; 2004]. At the same time, Denmark has other forms of distributed generation, e.g., extensive cogeneration of heat and power (CHP) plants for district heating or for covering industrial heat demands. This results in a high fuel-efficiency but also in a technically complex energy......Denmark has the World’s highest penetration of grid connected wind power in electricity generation with a share of 15.0% of total domestic demand in 2002 [Danish Energy Authority. Rapport fra arbejdsgruppen om kraftvarme- og VE-elektricitet. Bilagsrapport. Copenhagen: Danish Energy Authority; 2001...... as the potential impact of heat pumps used for district heating and installed for integration purposes. The analyses are made with particular focus on grid stability and delivery of ancillary services (required to control voltage and frequency) and demonstrate that it is possible to accommodate 50% or more wind...

  2. 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

    to be better understood. This relates both to the technical capabilities of the plants for delivering specific services and to the quantification of the needs. The paper presents the approach of the European IEE project REserviceS, aiming at establishing reference guidance for the ongoing developments......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...

  3. System design specification for the 1/4-scale tank and ancillary equipment

    International Nuclear Information System (INIS)

    Bamberger, J.A.; Bates, J.M.; Waters, E.D.; Heimberger, D.T.

    1993-09-01

    The Fluid Dynamic Test Facility (FDTF) is located in the 336 Building at the 300 Area of the Hanford Site. The FDTF will contain tanks that model the average internal diameter and height of a 3875 m 3 (1-million-gal) double-shell tank at both 1/12- and 1/4-scale, as well as ancillary equipment required to store, mix, and transport waste simulants. Experiments to be conducted in this facility will include investigations of sludge mobilization, slurry uniformity, aerosol generation, sludge washing, and instrumentation development to support start-up of the Hanford Waste Vitrification Project. This facility will also be used to model concepts and mitigating strategies to be used in the resolution of tank safety issues and the retrieval of waste from watch-list tanks

  4. Status of the segment interconnect, cable segment ancillary logic, and the cable segment hybrid driver projects

    International Nuclear Information System (INIS)

    Swoboda, C.; Barsotti, E.; Chappa, S.; Downing, R.; Goeransson, G.; Lensy, D.; Moore, G.; Rotolo, C.; Urish, J.

    1985-01-01

    The FASTBUS Segment Interconnect (SI) provides a communication path between two otherwise independent, asynchronous bus segments. In particular, the Segment Interconnect links a backplane crate segment to a cable segment. All standard FASTBUS address and data transactions can be passed through the SI or any number of SIs and segments in a path. Thus systems of arbitrary connection complexity can be formed, allowing simultaneous independent processing, yet still permitting devices associated with one segment to be accessed from others. The model S1 Segment Interconnect and the Cable Segment Ancillary Logic covered in this report comply with all the mandatory features stated in the FASTBUS specification document DOE/ER-0189. A block diagram of the SI is shown

  5. 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

  6. 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

  7. 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

    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 and ...

  8. 47 CFR 25.149 - Application requirements for ancillary terrestrial components in the mobile-satellite service...

    Science.gov (United States)

    2010-10-01

    ... 47 Telecommunication 2 2010-10-01 2010-10-01 false Application requirements for ancillary terrestrial components in the mobile-satellite service networks operating in the 1.5./1.6 GHz, 1.6/2.4 GHz and 2 GHz mobile-satellite service. 25.149 Section 25.149 Telecommunication FEDERAL COMMUNICATIONS...

  9. Profile of Hashimoto's Thyroiditis in Sri Lankans: Is There an Increased Risk of Ancillary Pathologies in Hashimoto's Thyroiditis?

    OpenAIRE

    Siriweera, Eranga Himalee; Ratnatunga, Neelakanthi Vajira Illangakoon

    2010-01-01

    Hashimoto's thyroiditis has been reported to be associated with many neoplastic and nonneoplastic thyroid pathologies. This retrospective study aims to determine the demographic profile of Hashimoto's thyroiditis in Sri Lankans, document ancillary pathologies in Hashimoto's thyroiditis, and determine whether there is an increased risk of occurrence of malignancies, benign neoplasms, and nonneoplastic benign lesions in Hashimoto's thyroiditis by comparing with thyroids showing multinodular goi...

  10. 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...

  11. Human aldolase B subunit-specific radioimmunoassay

    International Nuclear Information System (INIS)

    Asaka, M.; Alpert, E.

    1983-01-01

    A radioimmunoassay was developed for the direct quantification of aldolase B in human serum and tissues. The method is a double-antibody radioimmunoassay technique using radioiodinated aldolase B homopolymer as ligand, chicken antibodies to aldolase B and rabbit antibodies to chicken IgG. This radioimmunoassay was shown to be specific for the aldolase B subunit, with no cross-reactivity with either human aldolase A subunit or homopolymeric human aldolase C (C 4 ). The lowest measurable amount by this method was 2 ng/ml. Aldolase B is predominantly found in normal liver tissue, with relatively-high aldolase B levels also observed in kidney. Aldolase B levels in the serum obtained from 11 normal subjects ranged from 23 to 38 ng/ml, with a mean of 28.5 +- 9.2 (S.D.) ng/ml. Almost all of patients with hepatitis had serum aldolase B levels greater than 30 ng/ml. In cancer patients, serum aldolase B was slightly elevated in patients with metastatic liver cancer and primary lever cell carcinoma, whereas no elevation of serum aldolase B was shown in patients without liver metastasis. (Auth.)

  12. Subunit stoichiometry of the chloroplast photosystem I complex

    International Nuclear Information System (INIS)

    Bruce, B.D.; Malkin, R.

    1988-01-01

    A native photosystem I (PS I) complex and a PS I core complex depleted of antenna subunits has been isolated from the uniformly 14 C-labeled aquatic higher plant, Lemna. These complexes have been analyzed for their subunit stoichiometry by quantitative sodium dodecyl sulfate-polyacrylamide gel electrophoresis methods. The results for both preparations indicate that one copy of each high molecular mass subunit is present per PS I complex and that a single copy of most low molecular mass subunits is also present. These results suggest that iron-sulfur center X, an early PS I electron acceptor proposed to bind to the high molecular mass subunits, contains a single [4Fe-4S] cluster which is bound to a dimeric structure of high molecular mass subunits, each providing 2 cysteine residues to coordinate this cluster

  13. 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...

  14. 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

  15. 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

  16. Morphology combined with ancillary techniques: An algorithm approach for thyroid nodules.

    Science.gov (United States)

    Rossi, E D; Martini, M; Capodimonti, S; Cenci, T; Bilotta, M; Pierconti, F; Pontecorvi, A; Lombardi, C P; Fadda, G; Larocca, L M

    2018-04-23

    Several authors have underlined the limits of morphological analysis mostly in the diagnosis of follicular neoplasms (FN). The application of ancillary techniques, including immunocytochemistry (ICC) and molecular testing, contributes to a better definition of the risk of malignancy (ROM) and management of FN. According to literature, the application of models, including the evaluation of ICC, somatic mutations (ie, BRAF V 600E ), micro RNA analysis is proposed for FNs. This study discusses the validation of a diagnostic algorithm in FN with a special focus on the role of morphology then followed by ancillary techniques. From June 2014 to January 2016, we enrolled 37 FNs with histological follow-up. In the same reference period, 20 benign nodules and 20 positive for malignancy were selected as control. ICC, BRAF V 600E mutation and miR-375 were carried out on LBC. The 37 FNs included 14 atypia of undetermined significance/follicular lesion of undetermined significance and 23 FN. Specifically, atypia of undetermined significance/follicular lesion of undetermined significance resulted in three goitres, 10 follicular adenomas and one NIFTP whereas FN/suspicious for FN by seven follicular adenomas and 16 malignancies (nine non-invasive follicular thyroid neoplasms with papillary-like nuclear features, two invasive follicular variant of papillary thyroid carcinoma [PTC] and five PTC). The 20 positive for malignancy samples included two invasive follicular variant of PTC, 16 PTCs and two medullary carcinomas. The morphological features of BRAF V 600E mutation (nuclear features of PTC and moderate/abundant eosinophilic cytoplasms) were associated with 100% ROM. In the wild type cases, ROM was 83.3% in presence of a concordant positive ICC panel whilst significantly lower (10.5%) in a negative concordant ICC. High expression values of MirR-375 provided 100% ROM. The adoption of an algorithm might represent the best choice for the correct diagnosis of FNs. The morphological

  17. 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

    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......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...... of immunostimulators and antigens, respectively, into liposomes, and the choice of immunostimulator should ideally be based on knowledge regarding the specific PRR expression profile of the target APCs. Here, we review state-of-the-art formulation approaches employed for the inclusion of immunostimulators and subunit...

  18. 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.

  19. Assessing the economic potential of electric vehicles to provide ancillary services. The case of Germany

    Energy Technology Data Exchange (ETDEWEB)

    Schuller, Alexander; Rieger, Fabian [Karlsruhe Institute of Technology, Karlsruhe (Germany)

    2013-09-15

    The Vehicle-to-Grid (V2G) concept is a promising possibility for the integration of electric vehicles (EVs) into the power grid. This article presents an economic evaluation of EVs participating in the ancillary service market (primary, secondary and tertiary regulation) for the case of Germany, based on a price data set from 2011 and 2012. We examine the economic potential of nine general options to participate in the regulation market based on real-life EV specifications, connection powers and regulation energy prices. Results show that in the most profitable case a maximum average yearly profit of 730.31 Euro per vehicle is possible for negative regulation with payment direction TSO to provider in the secondary regulation market. Furthermore a sensitivity analysis is performed for all of the analyzed participation scenarios in order to identify crucial parameters for a possible V2G implementation. Major parameters for the successful implementation of V2G are the provided power per vehicle, the time an EV is available to the grid and the variable energy storage costs it incurs.

  20. 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.

  1. 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.

  2. Electric industry restructuring, ancillary services, and the potential impact on wind

    Energy Technology Data Exchange (ETDEWEB)

    Kirby, B.; Hirst, E. [Oak Ridge National Lab., TN (United States); Parsons, B.; Porter, K. [National Renewable Energy Lab., Golden, CO (United States)] [and others

    1997-12-31

    The new competitive electric power environment raises increased challenges for wind power. The DOE and EPRI wind programs have dealt extensively with the traditional vertically integrated utility planning and operating environment in which the host utility owns the generation (or purchases the power) and provides dispatch and transmission services. Under this traditional environment, 1794 MW of wind power, principally in California, have been successfully integrated into the U.S. electric power system. Another 4200 MW are installed elsewhere in the world. As issues have arisen, such as intermittency and voltage regulation, they have been successfully addressed with accepted power system procedures and practices. For an intermittent, non-dispatchable resource such as wind, new regulatory rules affecting power transmission services, raise questions about which ancillary services wind plants will be able to sell, which they will be required to purchase, and what the economic impacts will be on individual wind projects. This paper begins to look at issues of concern to wind in a restructured electric industry. The paper first briefly looks at the range of unbundled services and comments on their unique significance to wind. To illustrate the concerns that arise with restructuring, the paper then takes a more detailed look at a single service: regulation. Finally, the paper takes a brief look at technologies and strategies that could improve the competitive position of wind.

  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. A quality assurance program for ancillary high technology devices on a dual-energy accelerator

    International Nuclear Information System (INIS)

    Klein, Eric E.; Low, Daniel A.; Maag, Derek; Purdy, James A.

    1996-01-01

    Our facility has added high-technology ancillary devices to our dual-energy linear accelerator. After commissioning and acceptance testing of dual asymmetric jaws, dynamic wedge, portal imaging, and multileaf collimation (MLC), quality assurance programs were instituted. The programs were designed to be both periodic and patient specific when required. In addition, when dosimetric aspects were affected by these technologies, additional quality assurance checks were added. Positional accuracy checks (light and radiation) are done for both asymmetric jaws and MLC. Each patient MLC field is checked against the original simulation or digitally reconstructed radiographs. Off-axis factors and output checks are performed for asymmetric fields. Dynamic wedge transmission factors and profiles are checked periodically, and a patient diode check is performed for every new dynamic wedge portal. On-line imaging checks encompass safety checks along with periodic measurement of contrast and spatial resolution. The most important quality assurance activity is the annual review of proper operation and procedures for each device. Our programs have been successful in avoiding patient-related errors or device malfunctions. The programs are a team effort involving physicists, maintenance engineers, and therapists

  5. Prosomeric map of the lamprey forebrain based on calretinin immunocytochemistry, Nissl stain, and ancillary markers.

    Science.gov (United States)

    Pombal, M A; Puelles, L

    1999-11-22

    The structural organization of the lamprey extratelencephalic forebrain is re-examined from the perspective of the prosomeric segmental paradigm. The question asked was whether the prosomeric forebrain model used for gnathostomes is of material advantage for interpreting subdivisions in the lamprey forebrain. To this aim, the main longitudinal and transverse landmarks recognized by the prosomeric model in other vertebrates were identified in Nissl-stained lamprey material. Lines of cytoarchitectural discontinuity and contours of migrated neuronal groups were mapped in a two-dimensional sagittal representation and were also classified according to their radial position. Immunocytochemical mapping of calretinin expression in adjacent sections served to define particular structural units better, in particular, the dorsal thalamus. These data were complemented by numerous other chemoarchitectonic observations obtained with ancillary markers, which identified additional specific formations, subdivisions, or boundaries. Emphasis was placed on studying whether such chemically defined neuronal groups showed boundaries aligned with the postulated inter- or intraprosomeric boundaries. The course of diverse axonal tracts was studied also with regard to their prosomeric topography. This analysis showed that the full prosomeric model applies straightforwardly to the lamprey forebrain. This finding implies that a common segmental and longitudinal organization of the neural tube may be primitive for all vertebrates. Interesting novel aspects appear in the interpretation of the lamprey pretectum, the dorsal and ventral thalami, and the hypothalamus. The topologic continuity of the prosomeric forebrain regions with evaginated or non-evaginated portions of the telencephalon was also examined. Copyright 1999 Wiley-Liss, Inc.

  6. 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.

  7. 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.

  8. 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

  9. 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.

  10. Myristoylated α subunits of guanine nucleotide-binding regulatory proteins

    International Nuclear Information System (INIS)

    Buss, J.E.; Mumby, S.M.; Casey, P.J.; Gilman, A.G.; Sefton, B.M.

    1987-01-01

    Antisera directed against specific subunits of guanine nucleotide-binding regulatory proteins (G proteins) were used to immunoprecipitate these polypeptides from metabolically labeled cells. This technique detects, in extracts of a human astrocytoma cell line, the α subunits of G/sub s/ (stimulatory) (α 45 and α 52 ), a 41-kDa subunit of G/sub i/ (inhibitory) (α 41 ), a 40-kDa protein (α 40 ), and the 36-kDa β subunit. No protein that comigrated with the α subunit of G 0 (unknown function) (α 39 ) was detected. In cells grown in the presence of [ 3 H]myristic acid, α 41 and α 40 contained 3 H label, while the β subunit did not. Chemical analysis of lipids attached covalently to purified α 41 and α 39 from bovine brain also revealed myristic acid. Similar analysis of brain G protein β and γ subunits and of G/sub t/ (Transducin) subunits (α, β, and γ) failed to reveal fatty acids. The fatty acid associated with α 41 , α 40 , and α 39 was stable to treatment with base, suggesting that the lipid is linked to the polypeptide via an amide bond. These GTP binding proteins are thus identified as members of a select group of proteins that contains myristic acid covalently attached to the peptide backbone. Myristate may play an important role in stabilizing interactions of G proteins with phospholipid or with membrane-bound proteins

  11. 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. ... The sub-unit vaccine will be cost-effective, easy to produce, and safe. How it will make a ... IDRC invites applications for the IDRC Doctoral Research Awards.

  12. 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.

  13. 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.

  14. 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

  15. Studies on the subunits of human glycoprotein hormones in relation to reproduction

    International Nuclear Information System (INIS)

    Hagen, C.

    1977-01-01

    In this review summarising present knowledge of the biological and immunological activity of the subunits of human glycoprotein hormones, the specificity of the α-subunit and β-subunit radioimmunoassays are discussed. The crossreaction studies performed with the α-subunit radioimmunoassays are aummarised in one table while those with the β-subunit radioimmunoassays are presented in a second table. (JIW)

  16. 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

  17. Removal Action Work Plan for 105-DR and 105-F Building Interim Safe Storage Projects and Ancillary Buildings

    International Nuclear Information System (INIS)

    Morton, M.R.

    2000-01-01

    This document contains the removal action work plan 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, which is owned and operated by the US Department of Energy (DOE), in Benton County, Washington. The 100 Areas (including the 100-D/DR and 100-F Areas) of the Hanford Site were placed on the US Environmental Protection Agency's (EPA's) National Priorities List under the ''Comprehensive Environmental Response, Compensation, and Liability Act of 1980'' (CERCLA). The 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. The DOE has also determined that a non-time critical removal action is warranted at these facilities. Alternatives for conducting a non-time critical removal action were evaluated in the ''Engineering Evaluation/Cost Analysis for the 105-DR and 105-F Reactor Facilities and Ancillary Facilities'' (DOE-RL 1998a). The engineering evaluation/cost analysis (EE/CA) resulted in the recommendation to decontaminate and demolish the contaminated reactor buildings (except for the reactor blocks) and the ancillary facilities and to construct a safe storage enclosure (SSE) over the reactor blocks. The recommendation was approved in an action memorandum (Ecology et al. 1998) signed by the Washington State Department of Ecology (Ecology), EPA, and DOE. The DOE is the agency responsible for implementing the removal actions in the 105-D/DR and 105-F Areas. Ecology is the lead regulatory agency for facilities in the 100-D/DR Area, and EPA is the lead regulatory agency for facilities in the 100-F Area. The term ''lead regulator agency'' hereinafter, refers to these authorities. This removal action work plan supports implementation of the non-time critical removal action

  18. 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

  19. 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

  20. Allocation of reactive power support, active loss balancing and demand interruption ancillary services in MicroGrids

    Energy Technology Data Exchange (ETDEWEB)

    Gomes, Mario Helder [Departamento de Engenharia Electrotecnica, Instituto Politecnico de Tomar, Quinta do Contador, Estrada da Serra, 2300 Tomar (Portugal); Saraiva, Joao Tome [INESC Porto and Faculdade de Engenharia da Universidade do Porto, Campus da FEUP, Rua Dr. Roberto Frias, 4200 - 465 Porto (Portugal)

    2010-10-15

    MicroGrids represent a new paradigm for the operation of distribution systems and there are several advantages as well as challenges regarding their development. One of the advantages is related with the participation of MicroGrid agents in electricity markets and in the provision of ancillary services. This paper describes two optimization models to allocate three ancillary services among MicroGrid agents - reactive power/voltage control, active loss balancing and demand interruption. These models assume that MicroGrid agents participate in the day-ahead market sending their bids to the MicroGrid Central Controller, MGCC, that acts as an interface with the Market Operator. Once the Market Operator returns the economic dispatch of the MicroGrid agents, the MGCC checks its technical feasibility (namely voltage magnitude and branch flow limits) and activates an adjustment market to change the initial schedule and to allocate these three ancillary services. One of the models has crisp nature considering that voltage and branch flow limits are rigid while the second one admits that voltage and branch flow limits are modeled in a soft way using Fuzzy Set concepts. Finally, the paper illustrates the application of these models with a Case Study using a 55 node MV/LV network. (author)

  1. 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.

  2. Ancillary contributions of heterologous biotin protein ligase and carbonic anhydrase for CO2 incorporation into 3-hydroxypropionate by metabolically engineered Pyrococcus furiosus.

    Science.gov (United States)

    Lian, Hong; Zeldes, Benjamin M; Lipscomb, Gina L; Hawkins, Aaron B; Han, Yejun; Loder, Andrew J; Nishiyama, Declan; Adams, Michael W W; Kelly, Robert M

    2016-12-01

    Acetyl-Coenzyme A carboxylase (ACC), malonyl-CoA reductase (MCR), and malonic semialdehyde reductase (MRS) convert HCO 3 - and acetyl-CoA into 3-hydroxypropionate (3HP) in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation cycle resident in the extremely thermoacidophilic archaeon Metallosphaera sedula. These three enzymes, when introduced into the hyperthermophilic archaeon Pyrococcus furiosus, enable production of 3HP from maltose and CO 2 . Sub-optimal function of ACC was hypothesized to be limiting for production of 3HP, so accessory enzymes carbonic anhydrase (CA) and biotin protein ligase (BPL) from M. sedula were produced recombinantly in Escherichia coli to assess their function. P. furiosus lacks a native, functional CA, while the M. sedula CA (Msed_0390) has a specific activity comparable to other microbial versions of this enzyme. M. sedula BPL (Msed_2010) was shown to biotinylate the β-subunit (biotin carboxyl carrier protein) of the ACC in vitro. Since the native BPLs in E. coli and P. furiosus may not adequately biotinylate the M. sedula ACC, the carboxylase was produced in P. furiosus by co-expression with the M. sedula BPL. The baseline production strain, containing only the ACC, MCR, and MSR, grown in a CO 2 -sparged bioreactor reached titers of approximately 40 mg/L 3HP. Strains in which either the CA or BPL accessory enzyme from M. sedula was added to the pathway resulted in improved titers, 120 or 370 mg/L, respectively. The addition of both M. sedula CA and BPL, however, yielded intermediate titers of 3HP (240 mg/L), indicating that the effects of CA and BPL on the engineered 3HP pathway were not additive, possible reasons for which are discussed. While further efforts to improve 3HP production by regulating gene dosage, improving carbon flux and optimizing bioreactor operation are needed, these results illustrate the ancillary benefits of accessory enzymes for incorporating CO 2 into 3HP production in metabolically engineered P

  3. 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

  4. 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

  5. Histopathological diagnostic discrepancies in soft tissue tumours referred to a specialist centre: reassessment in the era of ancillary molecular diagnosis.

    Science.gov (United States)

    Thway, Khin; Wang, Jayson; Mubako, Taka; Fisher, Cyril

    2014-01-01

    Introduction. Soft tissue tumour pathology is a highly specialised area of surgical pathology, but soft tissue neoplasms can occur at virtually all sites and are therefore encountered by a wide population of surgical pathologists. Potential sarcomas require referral to specialist centres for review by pathologists who see a large number of soft tissue lesions and where appropriate ancillary investigations can be performed. We have previously assessed the types of diagnostic discrepancies between referring and final diagnosis for soft tissue lesions referred to our tertiary centre. We now reaudit this 6 years later, assessing changes in discrepancy patterns, particularly in relation to the now widespread use of ancillary molecular diagnostic techniques which were not prevalent in our original study. Materials and Methods. We compared the sarcoma unit's histopathology reports with referring reports on 348 specimens from 286 patients with suspected or proven soft tissue tumours in a one-year period. Results. Diagnostic agreement was seen in 250 cases (71.8%), with 57 (16.4%) major and 41 (11.8%) minor discrepancies. There were 23 cases of benign/malignant discrepancies (23.5% of all discrepancies). 50 ancillary molecular tests were performed, 33 for aiding diagnosis and 17 mutational analyses for gastrointestinal stromal tumour to guide therapy. Findings from ancillary techniques contributed to 3 major and 4 minor discrepancies. While the results were broadly similar to those of the previous study, there was an increase in frequency of major discrepancies. Conclusion. Six years following our previous study and notably now in an era of widespread ancillary molecular diagnosis, the overall discrepancy rate between referral and tertiary centre diagnosis remains similar, but there is an increase in frequency of major discrepancies likely to alter patient management. A possible reason for the increase in major discrepancies is the increasing lack of exposure to soft tissue

  6. The light subunit of system bo,+ is fully functional in the absence of the heavy subunit

    OpenAIRE

    Reig, Núria; Chillarón, Josep; Bartoccioni, Paola; Fernández, Esperanza; Bendahan, Annie; Zorzano, Antonio; Kanner, Baruch; Palacín, Manuel; Bertran, Joan

    2002-01-01

    The heteromeric amino acid transporters are composed of a type II glycoprotein and a non-glycosylated polytopic membrane protein. System bo,+ exchanges dibasic for neutral amino acids. It is composed of rBAT and bo,+AT, the latter being the polytopic membrane subunit. Mutations in either of them cause malfunction of the system, leading to cystinuria. bo,+AT-reconstituted systems from HeLa or MDCK cells catalysed transport of arginine that was totally dependent on the presence of one of the bo...

  7. Genetic analysis of the cytoplasmic dynein subunit families.

    Science.gov (United States)

    Pfister, K Kevin; Shah, Paresh R; Hummerich, Holger; Russ, Andreas; Cotton, James; Annuar, Azlina Ahmad; King, Stephen M; Fisher, Elizabeth M C

    2006-01-01

    Cytoplasmic dyneins, the principal microtubule minus-end-directed motor proteins of the cell, are involved in many essential cellular processes. The major form of this enzyme is a complex of at least six protein subunits, and in mammals all but one of the subunits are encoded by at least two genes. Here we review current knowledge concerning the subunits, their interactions, and their functional roles as derived from biochemical and genetic analyses. We also carried out extensive database searches to look for new genes and to clarify anomalies in the databases. Our analysis documents evolutionary relationships among the dynein subunits of mammals and other model organisms, and sheds new light on the role of this diverse group of proteins, highlighting the existence of two cytoplasmic dynein complexes with distinct cellular roles.

  8. Genetic analysis of the cytoplasmic dynein subunit families.

    Directory of Open Access Journals (Sweden)

    K Kevin Pfister

    2006-01-01

    Full Text Available Cytoplasmic dyneins, the principal microtubule minus-end-directed motor proteins of the cell, are involved in many essential cellular processes. The major form of this enzyme is a complex of at least six protein subunits, and in mammals all but one of the subunits are encoded by at least two genes. Here we review current knowledge concerning the subunits, their interactions, and their functional roles as derived from biochemical and genetic analyses. We also carried out extensive database searches to look for new genes and to clarify anomalies in the databases. Our analysis documents evolutionary relationships among the dynein subunits of mammals and other model organisms, and sheds new light on the role of this diverse group of proteins, highlighting the existence of two cytoplasmic dynein complexes with distinct cellular roles.

  9. The TadV Protein of Actinobacillus actinomycetemcomitans Is a Novel Aspartic Acid Prepilin Peptidase Required for Maturation of the Flp1 Pilin and TadE and TadF Pseudopilins†

    OpenAIRE

    Tomich, Mladen; Fine, Daniel H.; Figurski, David H.

    2006-01-01

    The tad locus of Actinobacillus actinomycetemcomitans encodes genes for the biogenesis of Flp pili, which allow the bacterium to adhere tenaciously to surfaces and form strong biofilms. Although tad (tight adherence) loci are widespread among bacterial and archaeal species, very little is known about the functions of the individual components of the Tad secretion apparatus. Here we characterize the mechanism by which the pre-Flp1 prepilin is processed to the mature pilus subunit. We demonstra...

  10. 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

  11. 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.

  12. 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

  13. 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.

  14. 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.

  15. 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

  16. Ancillary outcome measures for assessment of individuals with cervical spondylotic myelopathy.

    Science.gov (United States)

    Kalsi-Ryan, Sukhvinder; Singh, Anoushka; Massicotte, Eric M; Arnold, Paul M; Brodke, Darrel S; Norvell, Daniel C; Hermsmeyer, Jeffrey T; Fehlings, Michael G

    2013-10-15

    , Neck Disability Index, and 30-Meter Walk Test are most appropriate for the assessment of CSM. However, 6 additional outcome measures (QuickDASH, Berg Balance Scale, Graded Redefined Assessment of Strength Sensibility and Prehension, Grip Dynamometer, and GAITRite Analysis) were identified, which provide complementary assessments for CSM. SUMMARY STATEMENTS: There does not exist a single or composite of outcome instruments that measures myelopathy impairment, function/disability, and participation that have also demonstrated reliability, validity, and responsiveness in a CSM population. More work in the development and psychometric evaluation of new or existing measures is necessary to identify the ideal composite of measures to be used in the clinical and research settings. The mJOA, Nurick grade, NDI, MDI, and 30MWT should be adopted in any clinical practice that treats CSM both for screening and clinical follow-up. We propose that clinicians and researchers consider using the ancillary measures identified, such as the QuickDASH, Berg Balance Scale, GRASSP version 1.0, Grip Strength, and GAITRite Analysis. It is highly recommended that baseline and follow-up measurements should be performed in patients with CSM.

  17. 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.

  18. Ancillary care in public health intervention research in low-resource settings: researchers' practices and decision-making.

    Science.gov (United States)

    Taylor, Holly A; Merritt, Maria W; Mullany, Luke C

    2011-09-01

    Little is known about researchers' practices regarding the provision of ancillary care (AC) in public health intervention studies they have conducted and the factors that influence their decisions about whether to provide ancillary care in low-resource settings. We conducted 52 in-person in-depth interviews with public health researchers. Data analysis was iterative and led to the identification of themes and patterns among themes. We found that researchers who conduct their research in the community setting are more likely to identify and plan for the AC needs of potential research subjects before a study begins, whereas those affiliated with a permanent facility are more likely to deliver AC to research subjects on an ad hoc basis. Our findings suggest that on the whole, at least for public health intervention research in low-resource settings, researchers conducting research in the community setting confront more complex ethical and operational challenges in their decision-making about AC than do researchers conducting facility-based studies.

  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. A framework for cost-based pricing of transmission and ancillary services in competitive electric power markets

    International Nuclear Information System (INIS)

    Zobian, A.; Ilic, M.D.

    1995-01-01

    In this paper the authors propose a framework for accurate cost determination and pricing of transmission and ancillary services in competitive electric power markets. The proposed framework is based on their anticipation of the evolving environment and industry structure. They envision the future as a competitive energy market with a centralized control entity that coordinates system activities, prices transmission and ancillary services and controls various system resources. This control entity has control over a certain (pre-defined) geographical area. It is proposed that the system operation and control be kept as they are currently done in control centers, no major change in these functions is required for the proposed pricing strategy. The pricing strategy is divided into two main classes based on time scale separation and firmness, short and long term, firm and interruptible contracts. The approach is based on superposition of different transaction on the network, and a three-part tariff design. The charges are directly related to the impact of each transaction on the system

  1. The first transmembrane domain (TM1) of β2-subunit binds to the transmembrane domain S1 of α-subunit in BK potassium channels

    Science.gov (United States)

    Morera, Francisco J.; Alioua, Abderrahmane; Kundu, Pallob; Salazar, Marcelo; Gonzalez, Carlos; Martinez, Agustin D.; Stefani, Enrico; Toro, Ligia; Latorre, Ramon

    2012-01-01

    The BK channel is one of the most broadly expressed ion channels in mammals. In many tissues, the BK channel pore-forming α-subunit is associated to an auxiliary β-subunit that modulates the voltage- and Ca2+-dependent activation of the channel. Structural components present in β-subunits that are important for the physical association with the α-subunit are yet unknown. Here, we show through co-immunoprecipitation that the intracellular C-terminus, the second transmembrane domain (TM2) and the extracellular loop of the β2-subunit are dispensable for association with the α-subunit pointing transmembrane domain 1 (TM1) as responsible for the interaction. Indeed, the TOXCAT assay for transmembrane protein–protein interactions demonstrated for the first time that TM1 of the β2-subunit physically binds to the transmembrane S1 domain of the α-subunit. PMID:22710124

  2. 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

  3. Role of regulatory subunits and protein kinase inhibitor (PKI) in determining nuclear localization and activity of the catalytic subunit of protein kinase A.

    Science.gov (United States)

    Wiley, J C; Wailes, L A; Idzerda, R L; McKnight, G S

    1999-03-05

    Regulation of protein kinase A by subcellular localization may be critical to target catalytic subunits to specific substrates. We employed epitope-tagged catalytic subunit to correlate subcellular localization and gene-inducing activity in the presence of regulatory subunit or protein kinase inhibitor (PKI). Transiently expressed catalytic subunit distributed throughout the cell and induced gene expression. Co-expression of regulatory subunit or PKI blocked gene induction and prevented nuclear accumulation. A mutant PKI lacking the nuclear export signal blocked gene induction but not nuclear accumulation, demonstrating that nuclear export is not essential to inhibit gene induction. When the catalytic subunit was targeted to the nucleus with a nuclear localization signal, it was not sequestered in the cytoplasm by regulatory subunit, although its activity was completely inhibited. PKI redistributed the nuclear catalytic subunit to the cytoplasm and blocked gene induction, demonstrating that the nuclear export signal of PKI can override a strong nuclear localization signal. With increasing PKI, the export process appeared to saturate, resulting in the return of catalytic subunit to the nucleus. These results demonstrate that both the regulatory subunit and PKI are able to completely inhibit the gene-inducing activity of the catalytic subunit even when the catalytic subunit is forced to concentrate in the nuclear compartment.

  4. Pricing Energy and Ancillary Services in a Day-Ahead Market for a Price-Taker Hydro Generating Company Using a Risk-Constrained Approach

    Directory of Open Access Journals (Sweden)

    Perica Ilak

    2014-04-01

    Full Text Available This paper analyzes a price-taker hydro generating company which participates simultaneously in day-ahead energy and ancillary services markets. An approach for deriving marginal cost curves for energy and ancillary services is proposed, taking into consideration price uncertainty and opportunity cost of water, which can later be used to determine hourly bid curves. The proposed approach combines an hourly conditional value-at-risk, probability of occurrence of automatic generation control states and an opportunity cost of water to determine energy and ancillary services marginal cost curves. The proposed approach is in a linear constraint form and is easy to implement in optimization problems. A stochastic model of the hydro-economic river basin is presented, based on the actual Vinodol hydropower system in Croatia, with a complex three-dimensional relationship between the power produced, the discharged water, and the head of associated reservoir.

  5. 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).

  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. 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.

  8. 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.

  9. 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...... strong immune responses. This review highlights the status and recent advances in formulation and pulmonary delivery of thermostable human subunit vaccines. Such vaccines are very appealing from compliance, distribution and immunological point of view: Being non-invasive, inhalable vaccines are self...... immunity. Here, I review state of the art and perspectives in formulation design and processing methods for powder-based subunit vaccines intended for pulmonary administration, and present dry powder inhaler technologies suitable for translating these vaccines into clinical trials....

  10. 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

  11. 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

  12. 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

  13. Catalytic Subunit 1 of Protein Phosphatase 2A Is a Subunit of the STRIPAK Complex and Governs Fungal Sexual Development.

    Science.gov (United States)

    Beier, Anna; Teichert, Ines; Krisp, Christoph; Wolters, Dirk A; Kück, Ulrich

    2016-06-21

    The generation of complex three-dimensional structures is a key developmental step for most eukaryotic organisms. The details of the molecular machinery controlling this step remain to be determined. An excellent model system to study this general process is the generation of three-dimensional fruiting bodies in filamentous fungi like Sordaria macrospora Fruiting body development is controlled by subunits of the highly conserved striatin-interacting phosphatase and kinase (STRIPAK) complex, which has been described in organisms ranging from yeasts to humans. The highly conserved heterotrimeric protein phosphatase PP2A is a subunit of STRIPAK. Here, catalytic subunit 1 of PP2A was functionally characterized. The Δpp2Ac1 strain is sterile, unable to undergo hyphal fusion, and devoid of ascogonial septation. Further, PP2Ac1, together with STRIPAK subunit PRO22, governs vegetative and stress-related growth. We revealed in vitro catalytic activity of wild-type PP2Ac1, and our in vivo analysis showed that inactive PP2Ac1 blocks the complementation of the sterile deletion strain. Tandem affinity purification, followed by mass spectrometry and yeast two-hybrid analysis, verified that PP2Ac1 is a subunit of STRIPAK. Further, these data indicate links between the STRIPAK complex and other developmental signaling pathways, implying the presence of a large interconnected signaling network that controls eukaryotic developmental processes. The insights gained in our study can be transferred to higher eukaryotes and will be important for understanding eukaryotic cellular development in general. The striatin-interacting phosphatase and kinase (STRIPAK) complex is highly conserved from yeasts to humans and is an important regulator of numerous eukaryotic developmental processes, such as cellular signaling and cell development. Although functional insights into the STRIPAK complex are accumulating, the detailed molecular mechanisms of single subunits are only partially understood

  14. 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)

  15. The business of medicine: how to overcome financial obstacles and secure financing for your private practice and ancillary services business.

    Science.gov (United States)

    Nayor, David

    2012-01-01

    Doctors across the country who operate private medical practices are facing increasing financial obstacles, namely shrinking income as a result of rising costs and lower reimbursements. In addition, as hospitals have become overburdened many physicians have opened up private surgical centers; magnetic resonance imaging and computed tomography and positron emission tomography scanning facilities; pathology labs; colonoscopy/endoscopy suites; lithotripsy centers; and other medical businesses typically performed within the hospital. Moreover, many doctors seek loans to purchase existing practices or for their capital contribution in medical partnerships. The past decade has thus seen a significant increase in the number of doctors taking out small business loans. Indeed, banks view the healthcare industry as a large growth market. This article includes practical information, advice, and resources to help doctors to secure bank financing for their practices, ancillary services business, real estate, and equipment on the best possible market terms.

  16. 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)

  17. 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.

  18. 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

  19. 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.

  20. 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

  1. Partial agonists and subunit selectivity at NMDA receptors

    DEFF Research Database (Denmark)

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

    2010-01-01

    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...

  2. Therapeutic potential of Mediator complex subunits in metabolic diseases.

    Science.gov (United States)

    Ranjan, Amol; Ansari, Suraiya A

    2018-01-01

    The multisubunit Mediator is an evolutionary conserved transcriptional coregulatory complex in eukaryotes. It is needed for the transcriptional regulation of gene expression in general as well as in a gene specific manner. Mediator complex subunits interact with different transcription factors as well as components of RNA Pol II transcription initiation complex and in doing so act as a bridge between gene specific transcription factors and general Pol II transcription machinery. Specific interaction of various Mediator subunits with nuclear receptors (NRs) and other transcription factors involved in metabolism has been reported in different studies. Evidences indicate that ligand-activated NRs recruit Mediator complex for RNA Pol II-dependent gene transcription. These NRs have been explored as therapeutic targets in different metabolic diseases; however, they show side-effects as targets due to their overlapping involvement in different signaling pathways. Here we discuss the interaction of various Mediator subunits with transcription factors involved in metabolism and whether specific interaction of these transcription factors with Mediator subunits could be potentially utilized as therapeutic strategy in a variety of metabolic diseases. Copyright © 2017 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

  3. 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 ...

  4. Structural interaction of novel dendrimer and subunits with water

    African Journals Online (AJOL)

    Preferred Customer

    interaction study with solvents are essential [4-6] and several subunits are used for .... slowed down the viscous flow with higher excess limiting viscosities of the 2,4,6- ..... Practical Organic Chemistry, 5th ed.; Wiley: New York; 1989; p 300. 14.

  5. 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

  6. Moessbauer spectroscopic studies of hemoglobin and its isolated subunits

    International Nuclear Information System (INIS)

    Hoy, G.R.; Cook, D.C.; Berger, R.L.; Friedman, F.K.

    1986-01-01

    Samples of 90% enriched 57Fe hemoglobin and its isolated subunits have been prepared. Moessbauer spectroscopic measurements have been made on three such samples. Sample one contained contributions of oxyhemoglobin, deoxyhemoglobin, and carbonmonoxyhemoglobin. This sample was studied from a temperature of 90 K down to 230 mK. Measurements were also made at 4.2 K using a small applied magnetic field of 1.0 T. In general, the measured quadrupole splittings and isomer shifts for each component agreed with previous measurements on single component samples in the literature, and thus demonstrated that chemically enriched hemoglobin has not been altered. The second and third samples were isolated alpha and beta subunits, respectively. We have found measurable Moessbauer spectral differences between the HbO 2 sites in the alpha subunit sample and the beta subunit sample. The measured Moessbauer spectral areas indicate that the iron ion has the largest mean-square displacement at the deoxy Hb sites as compared to that at the oxy- and carbonmonoxy Hb sites. The mean-square displacement at the HbO 2 sites is the smallest

  7. 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...

  8. 47 CFR 25.254 - Special requirements for ancillary terrestrial components operating in the 1610-1626.5 MHz/2483.5...

    Science.gov (United States)

    2010-10-01

    ... 47 Telecommunication 2 2010-10-01 2010-10-01 false Special requirements for ancillary terrestrial components operating in the 1610-1626.5 MHz/2483.5-2500 MHz bands. 25.254 Section 25.254 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) COMMON CARRIER SERVICES SATELLITE COMMUNICATIONS Technical...

  9. 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.

  10. Catalytic Subunit 1 of Protein Phosphatase 2A Is a Subunit of the STRIPAK Complex and Governs Fungal Sexual Development

    Directory of Open Access Journals (Sweden)

    Anna Beier

    2016-06-01

    Full Text Available The generation of complex three-dimensional structures is a key developmental step for most eukaryotic organisms. The details of the molecular machinery controlling this step remain to be determined. An excellent model system to study this general process is the generation of three-dimensional fruiting bodies in filamentous fungi like Sordaria macrospora. Fruiting body development is controlled by subunits of the highly conserved striatin-interacting phosphatase and kinase (STRIPAK complex, which has been described in organisms ranging from yeasts to humans. The highly conserved heterotrimeric protein phosphatase PP2A is a subunit of STRIPAK. Here, catalytic subunit 1 of PP2A was functionally characterized. The Δpp2Ac1 strain is sterile, unable to undergo hyphal fusion, and devoid of ascogonial septation. Further, PP2Ac1, together with STRIPAK subunit PRO22, governs vegetative and stress-related growth. We revealed in vitro catalytic activity of wild-type PP2Ac1, and our in vivo analysis showed that inactive PP2Ac1 blocks the complementation of the sterile deletion strain. Tandem affinity purification, followed by mass spectrometry and yeast two-hybrid analysis, verified that PP2Ac1 is a subunit of STRIPAK. Further, these data indicate links between the STRIPAK complex and other developmental signaling pathways, implying the presence of a large interconnected signaling network that controls eukaryotic developmental processes. The insights gained in our study can be transferred to higher eukaryotes and will be important for understanding eukaryotic cellular development in general.

  11. Electrophysiology and Beyond: Multiple roles of Na+ channel β subunits in development and disease

    Science.gov (United States)

    Patino, Gustavo A.; Isom, Lori L.

    2010-01-01

    Voltage-gated Na+ channel (VGSC) β subunits are not “auxiliary.” These multifunctional molecules not only modulate Na+ current (INa), but also function as cell adhesion molecules (CAMs) – playing roles in aggregation, migration, invasion, neurite outgrowth, and axonal fasciculation. β subunits are integral members of VGSC signaling complexes at nodes of Ranvier, axon initial segments, and cardiac intercalated disks, regulating action potential propagation through critical intermolecular and cell-cell communication events. At least in vitro, many β subunit cell adhesive functions occur both in the presence and absence of pore-forming VGSC α subunits, and in vivo β subunits are expressed in excitable as well as non-excitable cells, thus β subunits may play important functional roles on their own, in the absence of α subunits. VGSC β1 subunits are essential for life and appear to be especially important during brain development. Mutations in β subunit genes result in a variety of human neurological and cardiovascular diseases. Moreover, some cancer cells exhibit alterations in β subunit expression during metastasis. In short, these proteins, originally thought of as merely accessory to α subunits, are critical players in their own right in human health and disease. Here we discuss the role of VGSC β subunits in the nervous system. PMID:20600605

  12. 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

  13. 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.

  14. 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

  15. 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...

  16. Mapping of the Mouse Actin Capping Protein Beta Subunit Gene

    Directory of Open Access Journals (Sweden)

    Cooper John A

    2000-07-01

    Full Text Available Abstract Background Capping protein (CP, a heterodimer of α and β subunits, is found in all eukaryotes. CP binds to the barbed ends of actin filaments in vitro and controls actin assembly and cell motility in vivo. Vertebrates have three isoforms of CPβ produced by alternatively splicing from one gene; lower organisms have one gene and one isoform. Results We isolated genomic clones corresponding to the β subunit of mouse CP and identified its chromosomal location by interspecies backcross mapping. Conclusions The CPβ gene (Cappb1 mapped to Chromosome 4 between Cdc42 and D4Mit312. Three mouse mutations, snubnose, curly tail, and cribriform degeneration, map in the vicinity of the β gene.

  17. 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.

  18. 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

  19. 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.

  20. Global Proteome Analysis Identifies Active Immunoproteasome Subunits in Human Platelets*

    Science.gov (United States)

    Klockenbusch, Cordula; Walsh, Geraldine M.; Brown, Lyda M.; Hoffman, Michael D.; Ignatchenko, Vladimir; Kislinger, Thomas; Kast, Juergen

    2014-01-01

    The discovery of new functions for platelets, particularly in inflammation and immunity, has expanded the role of these anucleate cell fragments beyond their primary hemostatic function. Here, four in-depth human platelet proteomic data sets were generated to explore potential new functions for platelets based on their protein content and this led to the identification of 2559 high confidence proteins. During a more detailed analysis, consistently high expression of the proteasome was discovered, and the composition and function of this complex, whose role in platelets has not been thoroughly investigated, was examined. Data set mining resulted in identification of nearly all members of the 26S proteasome in one or more data sets, except the β5 subunit. However, β5i, a component of the immunoproteasome, was identified. Biochemical analyses confirmed the presence of all catalytically active subunits of the standard 20S proteasome and immunoproteasome in human platelets, including β5, which was predominantly found in its precursor form. It was demonstrated that these components were assembled into the proteasome complex and that standard proteasome as well as immunoproteasome subunits were constitutively active in platelets. These findings suggest potential new roles for platelets in the immune system. For example, the immunoproteasome may be involved in major histocompatibility complex I (MHC I) peptide generation, as the MHC I machinery was also identified in our data sets. PMID:25146974

  1. Global proteome analysis identifies active immunoproteasome subunits in human platelets.

    Science.gov (United States)

    Klockenbusch, Cordula; Walsh, Geraldine M; Brown, Lyda M; Hoffman, Michael D; Ignatchenko, Vladimir; Kislinger, Thomas; Kast, Juergen

    2014-12-01

    The discovery of new functions for platelets, particularly in inflammation and immunity, has expanded the role of these anucleate cell fragments beyond their primary hemostatic function. Here, four in-depth human platelet proteomic data sets were generated to explore potential new functions for platelets based on their protein content and this led to the identification of 2559 high confidence proteins. During a more detailed analysis, consistently high expression of the proteasome was discovered, and the composition and function of this complex, whose role in platelets has not been thoroughly investigated, was examined. Data set mining resulted in identification of nearly all members of the 26S proteasome in one or more data sets, except the β5 subunit. However, β5i, a component of the immunoproteasome, was identified. Biochemical analyses confirmed the presence of all catalytically active subunits of the standard 20S proteasome and immunoproteasome in human platelets, including β5, which was predominantly found in its precursor form. It was demonstrated that these components were assembled into the proteasome complex and that standard proteasome as well as immunoproteasome subunits were constitutively active in platelets. These findings suggest potential new roles for platelets in the immune system. For example, the immunoproteasome may be involved in major histocompatibility complex I (MHC I) peptide generation, as the MHC I machinery was also identified in our data sets. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  2. 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

  3. The cytochrome oxidase subunit I and subunit III genes in Oenothera mitochondria are transcribed from identical promoter sequences

    Science.gov (United States)

    Hiesel, Rudolf; Schobel, Werner; Schuster, Wolfgang; Brennicke, Axel

    1987-01-01

    Two loci encoding subunit III of the cytochrome oxidase (COX) in Oenothera mitochondria have been identified from a cDNA library of mitochondrial transcripts. A 657-bp sequence block upstream from the open reading frame is also present in the two copies of the COX subunit I gene and is presumably involved in homologous sequence rearrangement. The proximal points of sequence rearrangements are located 3 bp upstream from the COX I and 1139 bp upstream from the COX III initiation codons. The 5'-termini of both COX I and COX III mRNAs have been mapped in this common sequence confining the promoter region for the Oenothera mitochondrial COX I and COX III genes to the homologous sequence block. ImagesFig. 5. PMID:15981332

  4. 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)

  5. 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

  6. 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 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.

  7. 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.

  8. 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-...

  9. 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.

  10. A Three-Part Electricity Price Mechanism for Photovoltaic-Battery Energy Storage Power Plants Considering the Power Quality and Ancillary Service

    Directory of Open Access Journals (Sweden)

    Yajing Gao

    2017-08-01

    Full Text Available To solve the problem of solar abandoning, which is accompanied by the rapid development of photovoltaic (PV power generation, a demonstration of a photovoltaic-battery energy storage system (PV-BESS power plant has been constructed in Qinghai province in China. However, it is difficult for the PV-BESS power plant to survive and develop with the current electricity price mechanism and subsidy policy. In this paper, a three-part electricity price mechanism is proposed based on a deep analysis of the construction and operation costs and economic income. The on-grid electricity price is divided into three parts: the capacity price, graded electricity price, and ancillary service price. First, to ensure that the investment of the PV-BESS power plant would achieve the industry benchmark income, the capacity price and benchmark electricity price are calculated using the discounted cash flow method. Then, the graded electricity price is calculated according to the grade of the quality of grid-connected power. Finally, the ancillary service price is calculated based on the graded electricity price and ancillary service compensation. The case studies verify the validity of the three-part electricity price mechanism. The verification shows that the three-part electricity price mechanism can help PV-BESS power plants to obtain good economic returns, which can promote the development of PV-BESS power plants.

  11. Molecular cloning of the human casein kinase II α subunit

    International Nuclear Information System (INIS)

    Meisner, H.; Heller-Harrison, R.; Buxton, J.; Czech, M.P.

    1989-01-01

    A human cDNA encoding the α subunit of casein kinase II and a partial cDNA encoding the rat homologue were isolated by using a Drosophila casein kinase II cDNA probe. The 2.2-kb human cDNA contains a 1.2-kb open reading frame, 150 nucleotides of 5' leader, and 850 nucleotides of 3' noncoding region. Except for the first 7 deduced amino acids that are missing in the rat cDNA, the 328 amino acids beginning with the amino terminus are identical between human and rat. The Drosophila enzyme sequence is 90% identical with the human casein kinase II sequence, and there is only a single amino acid difference between the published partial bovine sequence and the human sequence. In addition, the C-terminus of the human cDNA has an extra 53 amino acids not present in Drosophila. Northern analysis of rat and human RNA showed predominant bands of 5.5, 3.1, and 1.8 kb. In rat tissues, brain and spleen had the highest levels of casein kinase II α subunit specific RNA, while skeletal muscle showed the lowest. Southern analysis of human cultured cell and tissue genomic DNA using the full-length cDNA probe revealed two bands with restriction enzymes that have no recognition sites within the cDNA and three to six bands with enzymes having single internal sites. These results are consistent with the possibility that two genes encode the α subunits

  12. Covalent dimerization of ribulose bisphosphate carboxylase subunits by UV radiation

    Energy Technology Data Exchange (ETDEWEB)

    Ferreira, R.M.B. [Universidade Tecnica, Lisbon (Portugal). Inst. Superior de Agronomia]|[Universidade Nova de Lisboa, Oeiras (Portugal). Instituto de Tecnologia Quimica e Biologica; Franco, E.; Teixeira, A.R.N. [Universidade Tecnica, Lisbon (Portugal). Inst. Superior de Agronomia

    1996-08-15

    The effect of UV radiation (UV-A, UV-B and UV-C) on ribulose bisphosphate carboxylase from a variety of plant species was examined. The exposition of plant leaves or the pure enzyme to UV radiation produced a UV-dependent accumulation of a 65 kDa polypeptide (P65). Different approaches were utilized to elucidate the origin and structure of P65: electrophoretic and fluorographic analyses of {sup 35}S-labelled ribulose biphosphate carboxylase exposed to UV radiation and immunological experiments using antibodies specific for P65, for the large and small subunits of ribulose biphosphate carboxylase and for high-molecular-mass aggregates of the enzyme. These studies revealed that P65 is a dimer, formed by the covalent, non-disulphide linkage of one small subunit with one large subunit of ribulose biphosphate carboxylase. For short periods of time (<1 h), the amount of P65 formed increased with the duration of the exposure to the UV radiation and with the energy of the radiation applied. Prolonged exposure to UV radiation (1-6 h) resulted in the formation of high-molecular-mass aggregates of ribulose biphosphate carboxylase. Formation of P65 was shown to depend on the native state of the protein, was stimulated by inhibitors of enzyme activity, and was inhibited by activators of enzyme activity. A UV-independent accumulation of P65 was also achieved by the in vitro incubation of plant crude extracts. However, the UV-dependent and the UV-independent formation of P65 seemed to occur by distinct molecular mechanisms. The UV-dependent accumulation of P65 was immunologically detected in all species examined, including Lemna minor, Arum italicum, Brassica oleracea, Triticum aestivum, Zea mays, Pisum sativum and Phaseolus vulgaris, suggesting that it may constitute a universal response to UV radiation, common to all photosynthetic tissues. (Author).

  13. Covalent dimerization of ribulose bisphosphate carboxylase subunits by UV radiation

    International Nuclear Information System (INIS)

    Ferreira, R.M.B.; Universidade Nova de Lisboa, Oeiras; Franco, E.; Teixeira, A.R.N.

    1996-01-01

    The effect of UV radiation (UV-A, UV-B and UV-C) on ribulose bisphosphate carboxylase from a variety of plant species was examined. The exposition of plant leaves or the pure enzyme to UV radiation produced a UV-dependent accumulation of a 65 kDa polypeptide (P65). Different approaches were utilized to elucidate the origin and structure of P65: electrophoretic and fluorographic analyses of 35 S-labelled ribulose biphosphate carboxylase exposed to UV radiation and immunological experiments using antibodies specific for P65, for the large and small subunits of ribulose biphosphate carboxylase and for high-molecular-mass aggregates of the enzyme. These studies revealed that P65 is a dimer, formed by the covalent, non-disulphide linkage of one small subunit with one large subunit of ribulose biphosphate carboxylase. For short periods of time (<1 h), the amount of P65 formed increased with the duration of the exposure to the UV radiation and with the energy of the radiation applied. Prolonged exposure to UV radiation (1-6 h) resulted in the formation of high-molecular-mass aggregates of ribulose biphosphate carboxylase. Formation of P65 was shown to depend on the native state of the protein, was stimulated by inhibitors of enzyme activity, and was inhibited by activators of enzyme activity. A UV-independent accumulation of P65 was also achieved by the in vitro incubation of plant crude extracts. However, the UV-dependent and the UV-independent formation of P65 seemed to occur by distinct molecular mechanisms. The UV-dependent accumulation of P65 was immunologically detected in all species examined, including Lemna minor, Arum italicum, Brassica oleracea, Triticum aestivum, Zea mays, Pisum sativum and Phaseolus vulgaris, suggesting that it may constitute a universal response to UV radiation, common to all photosynthetic tissues. (Author)

  14. Flexible Connectors between Capsomer Subunits that Regulate Capsid Assembly.

    Science.gov (United States)

    Hasek, Mary L; Maurer, Joshua B; Hendrix, Roger W; Duda, Robert L

    2017-08-04

    Viruses build icosahedral capsids of specific size and shape by regulating the spatial arrangement of the hexameric and pentameric protein capsomers in the growing shell during assembly. In the T=7 capsids of Escherichia coli bacteriophage HK97 and other phages, 60 capsomers are hexons, while the rest are pentons that are correctly positioned during assembly. Assembly of the HK97 capsid to the correct size and shape has been shown to depend on specific ionic contacts between capsomers. We now describe additional ionic interactions within capsomers that also regulate assembly. Each is between the long hairpin, the "E-loop," that extends from one subunit to the adjacent subunit within the same capsomer. Glutamate E153 on the E-loop and arginine R210 on the adjacent subunit's backbone alpha-helix form salt bridges in hexamers and pentamers. Mutations that disrupt these salt bridges were lethal for virus production, because the mutant proteins assembled into tubes or sheets instead of capsids. X-ray structures show that the E153-R210 links are flexible and maintained during maturation despite radical changes in capsomer shape. The E153-R210 links appear to form early in assembly to enable capsomers to make programmed changes in their shape during assembly. The links also prevent flattening of capsomers and premature maturation. Mutant phenotypes and modeling support an assembly model in which flexible E153-R210 links mediate capsomer shape changes that control where pentons are placed to create normal-sized capsids. The E-loop may be conserved in other systems in order to play similar roles in regulating assembly. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Sex Hormone Receptor Expression in the Human Vocal Fold Subunits.

    Science.gov (United States)

    Kirgezen, Tolga; Sunter, Ahmet Volkan; Yigit, Ozgur; Huq, Gulben Erdem

    2017-07-01

    The study aimed to evaluate the existence of sex hormone receptors in the subunits of vocal fold. This is a cadaver study. The androgen, estrogen, and progesterone receptors were examined in the epithelium (EP), superficial layer of the lamina propria (SLP), vocal ligament (VL), and macula flava (MF) of the vocal folds from 42 human cadavers (21 male, 21 female) by immunohistochemical methods. Their staining ratios were scored and statistically compared. The androgen receptor score was significantly higher for the MF than for the EP and SLP (P vocal fold, mostly in the MF and VLs. Copyright © 2017 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

  16. 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.

  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. 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.

  20. A charged residue at the subunit interface of PCNA promotes trimer formation by destabilizing alternate subunit interactions

    International Nuclear Information System (INIS)

    Freudenthal, Bret D.; Gakhar, Lokesh; Ramaswamy, S.; Washington, M. Todd

    2009-01-01

    Eukaryotic proliferating cell nuclear antigen (PCNA), an essential accessory factor in DNA replication and repair, is a ring-shaped homotrimer. A novel nontrimeric structure of E113G-mutant PCNA protein is reported, which shows that this protein forms alternate subunit interactions. It is concluded that the charged side chain of Glu113 promotes normal trimer formation by destabilizing these alternate subunit interactions. Eukaryotic proliferating cell nuclear antigen (PCNA) is an essential replication accessory factor that interacts with a variety of proteins involved in DNA replication and repair. Each monomer of PCNA has an N-terminal domain A and a C-terminal domain B. In the structure of the wild-type PCNA protein, domain A of one monomer interacts with domain B of a neighboring monomer to form a ring-shaped trimer. Glu113 is a conserved residue at the subunit interface in domain A. Two distinct X-ray crystal structures have been determined of a mutant form of PCNA with a substitution at this position (E113G) that has previously been studied because of its effect on translesion synthesis. The first structure was the expected ring-shaped trimer. The second structure was an unanticipated nontrimeric form of the protein. In this nontrimeric form, domain A of one PCNA monomer interacts with domain A of a neighboring monomer, while domain B of this monomer interacts with domain B of a different neighboring monomer. The B–B interface is stabilized by an antiparallel β-sheet and appears to be structurally similar to the A–B interface observed in the trimeric form of PCNA. The A–A interface, in contrast, is primarily stabilized by hydrophobic interactions. Because the E113G substitution is located on this hydrophobic surface, the A–A interface should be less favorable in the case of the wild-type protein. This suggests that the side chain of Glu113 promotes trimer formation by destabilizing these possible alternate subunit interactions

  1. Determinants of RNA polymerase alpha subunit for interaction with beta, beta', and sigma subunits: hydroxyl-radical protein footprinting.

    OpenAIRE

    Heyduk, T; Heyduk, E; Severinov, K; Tang, H; Ebright, R H

    1996-01-01

    Escherichia coli RNA polymerase (RNAP) alpha subunit serves as the initiator for RNAP assembly, which proceeds according to the pathway 2 alpha-->alpha 2-->alpha 2 beta-->alpha 2 beta beta'-->alpha 2 beta beta' sigma. In this work, we have used hydroxyl-radical protein footprinting to define determinants of alpha for interaction with beta, beta', and sigma. Our results indicate that amino acids 30-75 of alpha are protected from hydroxyl-radical-mediated proteolysis upon interaction with beta ...

  2. Heterotrimeric G protein subunits are located on rat liver endosomes

    Directory of Open Access Journals (Sweden)

    Van Dyke Rebecca W

    2004-01-01

    Full Text Available Abstract Background Rat liver endosomes contain activated insulin receptors and downstream signal transduction molecules. We undertook these studies to determine whether endosomes also contain heterotrimeric G proteins that may be involved in signal transduction from G protein-coupled receptors. Results By Western blotting Gsα, Giα1,2, Giα3 and Gβ were enriched in both canalicular (CM and basolateral (BLM membranes but also readily detectable on three types of purified rat liver endosomes in the order recycling receptor compartment (RRC > compartment for uncoupling of receptor and ligand (CURL > multivesicular bodies (MVB >> purified secondary lysosomes. Western blotting with antibodies to Na, K-ATPase and to other proteins associated with plasma membranes and intracellular organelles indicated this was not due to contamination of endosome preparations by CM or BLM. Adenylate cyclase (AC was also identified on purified CM, BLM, RRC, CURL and MVB. Percoll gradient fractionation of liver postnuclear supernatants demonstrated co-occurrence of endosomes and heterotrimeric G protein subunits in fractions with little plasma membrane markers. By confocal microscopy, punctate staining for Gsα, Giα3 and Gβ corresponded to punctate areas of endocytosed Texas red-dextran in hepatocytes from control and cholera toxin-treated livers. Conclusion We conclude that heterotrimeric G protein subunits as well as AC likely traffic into hepatocytes on endosome membranes, possibly generating downstream signals spatially separate from signalling generated at the plasma membrane, analogous to the role(s of internalized insulin receptors.

  3. Fungal mediator tail subunits contain classical transcriptional activation domains.

    Science.gov (United States)

    Liu, Zhongle; Myers, Lawrence C

    2015-04-01

    Classical activation domains within DNA-bound eukaryotic transcription factors make weak interactions with coactivator complexes, such as Mediator, to stimulate transcription. How these interactions stimulate transcription, however, is unknown. The activation of reporter genes by artificial fusion of Mediator subunits to DNA binding domains that bind to their promoters has been cited as evidence that the primary role of activators is simply to recruit Mediator. We have identified potent classical transcriptional activation domains in the C termini of several tail module subunits of Saccharomyces cerevisiae, Candida albicans, and Candida dubliniensis Mediator, while their N-terminal domains are necessary and sufficient for their incorporation into Mediator but do not possess the ability to activate transcription when fused to a DNA binding domain. This suggests that Mediator fusion proteins actually are functioning in a manner similar to that of a classical DNA-bound activator rather than just recruiting Mediator. Our finding that deletion of the activation domains of S. cerevisiae Med2 and Med3, as well as C. dubliniensis Tlo1 (a Med2 ortholog), impairs the induction of certain genes shows these domains function at native promoters. Activation domains within coactivators are likely an important feature of these complexes and one that may have been uniquely leveraged by a common fungal pathogen. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  4. 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.

  5. 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

  6. 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.

  7. Role of the beta subunit of casein kinase-2 on the stability and specificity of the recombinant reconstituted holoenzyme

    DEFF Research Database (Denmark)

    Meggio, F; Boldyreff, B; Marin, O

    1992-01-01

    Recombinant human alpha subunit from casein kinase-2 (CK-2) was subjected, either alone or in combination with recombinant human beta subunit, to high temperature, tryptic digestion and urea treatment. In all three cases, it was shown that the presence of the beta subunit could drastically reduce...... the autophosphorylation site. It is suggested that the acidic domain of the beta subunit, encompassing residues 55-71, plays a role in the interactions between the beta and alpha subunits....

  8. 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.

  9. 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

  10. Antibodies to the α-subunit of insulin receptor from eggs of immunized hens

    International Nuclear Information System (INIS)

    Song, C.; Yu, J.; Bai, D.H.; Hester, P.Y.; Kim, K.

    1985-01-01

    Simple methods for the generation, purification, and assay of antibodies to the α-subunit of insulin receptor from eggs of immunized hen have been described. Chicken antibodies against the α-subunit inhibit insulin binding to the receptor and stimulate glucose oxidation as well as autophosphorylation of the β-subunit. Thus the properties of chicken antibodies are very similar to those of antibodies found in human autoimmune diseases and different from rabbit antibodies obtained against the same antigen

  11. 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....

  12. 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...

  13. Subunits of the Snf1 kinase heterotrimer show interdependence for association and activity.

    Science.gov (United States)

    Elbing, Karin; Rubenstein, Eric M; McCartney, Rhonda R; Schmidt, Martin C

    2006-09-08

    The Snf1 kinase and its mammalian orthologue, the AMP-activated protein kinase (AMPK), function as heterotrimers composed of a catalytic alpha-subunit and two non-catalytic subunits, beta and gamma. The beta-subunit is thought to hold the complex together and control subcellular localization whereas the gamma-subunit plays a regulatory role by binding to and blocking the function of an auto-inhibitory domain (AID) present in the alpha-subunit. In addition, catalytic activity requires phosphorylation by a distinct upstream kinase. In yeast, any one of three Snf1-activating kinases, Sak1, Tos3, or Elm1, can fulfill this role. We have previously shown that Sak1 is the only Snf1-activating kinase that forms a stable complex with Snf1. Here we show that the formation of the Sak1.Snf1 complex requires the beta- and gamma-subunits in vivo. However, formation of the Sak1.Snf1 complex is not necessary for glucose-regulated phosphorylation of the Snf1 activation loop. Snf1 kinase purified from cells lacking the beta-subunits do not contain any gamma-subunit, indicating that the Snf1 kinase does not form a stable alphagamma dimer in vivo. In vitro kinase assays using purified full-length and truncated Snf1 proteins demonstrate that the kinase domain, which lacks the AID, is significantly more active than the full-length Snf1 protein. Addition of purified beta- and gamma-subunits could stimulate the kinase activity of the full-length alpha-subunit but only when all three subunits were present, suggesting an interdependence of all three subunits for assembly of a functional complex.

  14. The testis-specific Cα2 subunit of PKA is kinetically indistinguishable from the common Cα1 subunit of PKA

    Directory of Open Access Journals (Sweden)

    Herberg Friedrich W

    2011-08-01

    Full Text Available Abstract Background The two variants of the α-form of the catalytic (C subunit of protein kinase A (PKA, designated Cα1 and Cα2, are encoded by the PRKACA gene. Whereas Cα1 is ubiquitous, Cα2 expression is restricted to the sperm cell. Cα1 and Cα2 are encoded with different N-terminal domains. In Cα1 but not Cα2 the N-terminal end introduces three sites for posttranslational modifications which include myristylation at Gly1, Asp-specific deamidation at Asn2 and autophosphorylation at Ser10. Previous reports have implicated specific biological features correlating with these modifications on Cα1. Since Cα2 is not modified in the same way as Cα1 we tested if they have distinct biochemical activities that may be reflected in different biological properties. Results We show that Cα2 interacts with the two major forms of the regulatory subunit (R of PKA, RI and RII, to form cAMP-sensitive PKAI and PKAII holoenzymes both in vitro and in vivo as is also the case with Cα1. Moreover, using Surface Plasmon Resonance (SPR, we show that the interaction patterns of the physiological inhibitors RI, RII and PKI were comparable for Cα2 and Cα1. This is also the case for their potency to inhibit catalytic activities of Cα2 and Cα1. Conclusion We conclude that the regulatory complexes formed with either Cα1 or Cα2, respectively, are indistinguishable.

  15. 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.

  16. Distinct forms of the β subunit of GTP-binding regulatory proteins identified by molecular cloning

    International Nuclear Information System (INIS)

    Fong, H.K.W.; Amatruda, T.T. III; Birren, B.W.; Simon, M.I.

    1987-01-01

    Two distinct β subunits of guanine nucleotide-binding regulatory proteins have been identified by cDNA cloning and are referred to as β 1 and β 1 subunits. The bovine transducin β subunit (β 1 ) has been cloned previously. The author now isolated and analyzed cDNA clones that encode the β 2 subunit from bovine adrenal, bovine brain, and a human myeloid leukemia cell line, HL-60. The 340-residue M/sub r/ 37,329 Β 2 protein is 90% identical with β 1 in predicted amino acid sequence, and it is also organized as a series of repetitive homologous segments. The major mRNA that encodes the bovine β 2 subunit is 1.7 kilobases in length. It is expressed at lower levels than β 1 subunit mRNA in all tissues examined. The β 1 and β 2 messages are expressed in cloned human cell lines. Hybridization of cDNA probes to bovine DNA showed that β 1 and β 2 are encoded by separate genes. The amino acid sequences for the bovine and human β 2 subunit are identical, as are the amino acid sequences for the bovine and human β 1 subunit. This evolutionary conservation suggests that the two β subunits have different roles in the signal transduction process

  17. 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.

  18. 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

  19. Immunochemical analysis of Micrococcus lysodeikticus (luteus) F1-ATPase and its subunits.

    Science.gov (United States)

    Urban, C; Salton, M R

    1983-08-31

    The F1-ATPase from Micrococcus lysodeikticus has been purified to 95% protein homogeneity in this laboratory and as all other bacterial F1S, possesses five distinct subunits with molecular weights ranging from 60 000 to 10 000 (Huberman, M. and Salton, M.R.J. (1979) Biochim. Biophys. Acta 547, 230-240). In this communication, we demonstrate the immunochemical reactivities of antibodies to native and SDS-dissociated subunits with the native and dissociated F1-ATPase and show that: (1) the antibodies generated to the native or SDS-dissociated subunits react with the native molecule; (2) all of the subunits comprising the F1 are antigenically unique as determined by crossed immunoelectrophoresis and the Ouchterlony double-diffusion techniques; (3) antibodies to the SDS-denatured individual delta- and epsilon-subunits can be used to destabilize the interaction of these specific subunits with the rest of the native F1; and (4) all subunit antibodies as well as anti-native F1 were found to inhibit ATPase activity to varying degrees, the strongest inhibition being seen with antibodies to the total F1 and anti-alpha- and anti-beta-subunit antibodies. The interaction of specific subunit antibodies may provide a new and novel way to study further and characterize the catalytic portions of F1-ATPases and in general may offer an additional method for the examination of multimeric proteins.

  20. Effect of HMM Glutenin Subunits on Wheat Quality Attributes

    Directory of Open Access Journals (Sweden)

    Daniela Horvat

    2009-01-01

    Full Text Available Glutenin is a group of polymeric gluten proteins. Glutenin molecules consist of glutenin subunits linked together with disulphide bonds and having higher (HMM-GS and lower (LMM-GS molecular mass. The main objective of this study is the evaluation of the influence of HMM-GS on flour processing properties. Seven bread wheat genotypes with contrasting quality attributes and different HMM-GS composition were analyzed during three years. The composition and quantity of HMM-GS were determined by SDS-PAGE and RP-HPLC, respectively. The quality diversity among genotypes was estimated by the analysis of wheat grain, and flour and bread quality parameters. The presence of HMM glutenin subunits 1 and 2* at Glu-A1 and the subunits 5+10 at Glu-D1 loci, as well as a higher proportion of total HMM-GS, had a positive effect on wheat quality. Cluster analysis of the three groups of data (genotype and HMM-GS, flour and bread quality, and dough rheology yielded the same hierarchical structure for the first top three levels, and similarity of the corresponding dendrograms was proved by the principal eigenvalues of the corresponding Euclidian distance matrices. The obtained similarity in classification based on essentially different types of measurements reflects strong natural association between genetic data, product quality and physical properties. Principal component analysis (PCA was applied to effectively reduce large data set into lower dimensions of latent variables amenable for the analysis. PCA analysis of the total set of data (15 variables revealed a very strong interrelationship between the variables. The first three PCA components accounted for 96 % of the total variance, which was significant to the level of 0.05 and was considered as the level of experimental error. These data imply that the quality of wheat cultivars can be contributed to HMM-GS data and should be taken into account in breeding programs assisted by computer models with the aim to

  1. 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

  2. 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.

  3. 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.

  4. 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)

  5. Differential regulation of thyrotropin subunit apoprotein and carbohydrate biosynthesis by thyroid hormone

    International Nuclear Information System (INIS)

    Taylor, T.; Weintraub, B.D.

    1985-01-01

    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 [ 14 C]alanine and [ 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, [ 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. [ 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

  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. Potential of vehicle-to-grid ancillary services considering the uncertainties in plug-in electric vehicle availability and service/localization limitations in distribution grids

    International Nuclear Information System (INIS)

    Sarabi, Siyamak; Davigny, Arnaud; Courtecuisse, Vincent; Riffonneau, Yann; Robyns, Benoît

    2016-01-01

    Highlights: • The availability uncertainty of PEVs are modelled using Gaussian mixture model. • Interdependency of stochastic variables are modelled using copula function. • V2G bidding capacity is calculated using Free Pattern search optimization method. • Localization limitation is considered for V2G service potential assessment. • Competitive services for fleet of V2G-enabled PEVs are identified using fuzzy sets. - Abstract: The aim of the paper is to propose an approach for statistical assessment of the potential of plug-in electric vehicles (PEV) for vehicle-to-grid (V2G) ancillary services, where it focuses on PEVs doing daily home-work commuting. In this approach, the possible ancillary services (A/S) for each PEV fleet in terms of its available V2G power (AVP) and flexible intervals are identified. The flexible interval is calculated using a powerful stochastic global optimization technique so-called “Free Pattern Search” (FPS). A probabilistic method is also proposed to quantify the impacts of PEV’s availability uncertainty using the Gaussian mixture model (GMM), and interdependency of stochastic variables on AVP of each fleet thanks to a multivariate modeling with Copula function. Each fleet is analyzed based on its aggregated PEV numbers at different level of distribution grid, in order to satisfy the ancillary services localization limitation. A case study using the proposed approach evaluates the real potential in Niort, a city in west of France. In fact, by using the proposed approach an aggregator can analyze the V2G potential of PEVs under its contract.

  8. Compilation, quality control, analysis, and summary of discrete suspended-sediment and ancillary data in the United States, 1901-2010

    Science.gov (United States)

    Lee, Casey J.; Glysson, G. Douglas

    2013-01-01

    Human-induced and natural changes to the transport of sediment and sediment-associated constituents can degrade aquatic ecosystems and limit human uses of streams and rivers. The lack of a dedicated, easily accessible, quality-controlled database of sediment and ancillary data has made it difficult to identify sediment-related water-quality impairments and has limited understanding of how human actions affect suspended-sediment concentrations and transport. The purpose of this report is to describe the creation of a quality-controlled U.S. Geological Survey suspended-sediment database, provide guidance for its use, and summarize characteristics of suspended-sediment data through 2010. The database is provided as an online application at http://cida.usgs.gov/sediment to allow users to view, filter, and retrieve available suspended-sediment and ancillary data. A data recovery, filtration, and quality-control process was performed to expand the availability, representativeness, and utility of existing suspended-sediment data collected by the U.S. Geological Survey in the United States before January 1, 2011. Information on streamflow condition, sediment grain size, and upstream landscape condition were matched to sediment data and sediment-sampling sites to place data in context with factors that may influence sediment transport. Suspended-sediment and selected ancillary data are presented from across the United States with respect to time, streamflow, and landscape condition. Examples of potential uses of this database for identifying sediment-related impairments, assessing trends, and designing new data collection activities are provided. This report and database can support local and national-level decision making, project planning, and data mining activities related to the transport of suspended-sediment and sediment-associated constituents.

  9. 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.

  10. 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.

  11. 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.

  12. 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

  13. Differential expression of BK channel isoforms and beta-subunits in rat neuro-vascular tissues

    DEFF Research Database (Denmark)

    Poulsen, Asser Nyander; Wulf, Helle; Hay-Schmidt, Anders

    2009-01-01

    We investigated the expression of splice variants and beta-subunits of the BK channel (big conductance Ca(2+)-activated K(+) channel, Slo1, MaxiK, K(Ca)1.1) in rat cerebral blood vessels, meninges, trigeminal ganglion among other tissues. An alpha-subunit splice variant X1(+24) was found expresse...

  14. 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.

  15. 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...

  16. 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.

  17. 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...

  18. 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

  19. 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.

  20. 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.

  1. Mining Protein Evolution for Insights into Mechanisms of Voltage-Dependent Sodium Channel Auxiliary Subunits.

    Science.gov (United States)

    Molinarolo, Steven; Granata, Daniele; Carnevale, Vincenzo; Ahern, Christopher A

    2018-02-21

    Voltage-gated sodium channel (VGSC) beta (β) subunits have been called the "overachieving" auxiliary ion channel subunit. Indeed, these subunits regulate the trafficking of the sodium channel complex at the plasma membrane and simultaneously tune the voltage-dependent properties of the pore-forming alpha-subunit. It is now known that VGSC β-subunits are capable of similar modulation of multiple isoforms of related voltage-gated potassium channels, suggesting that their abilities extend into the broader voltage-gated channels. The gene family for these single transmembrane immunoglobulin beta-fold proteins extends well beyond the traditional VGSC β1-β4 subunit designation, with deep roots into the cell adhesion protein family and myelin-related proteins - where inherited mutations result in a myriad of electrical signaling disorders. Yet, very little is known about how VGSC β-subunits support protein trafficking pathways, the basis for their modulation of voltage-dependent gating, and, ultimately, their role in shaping neuronal excitability. An evolutionary approach can be useful in yielding new clues to such functions as it provides an unbiased assessment of protein residues, folds, and functions. An approach is described here which indicates the greater emergence of the modern β-subunits roughly 400 million years ago in the early neurons of Bilateria and bony fish, and the unexpected presence of distant homologues in bacteriophages. Recent structural breakthroughs containing α and β eukaryotic sodium channels containing subunits suggest a novel role for a highly conserved polar contact that occurs within the transmembrane segments. Overall, a mixture of approaches will ultimately advance our understanding of the mechanism for β-subunit interactions with voltage-sensor containing ion channels and membrane proteins.

  2. Topographic antigenic determinants recognized by monoclonal antibodies on human choriogonadotropin beta-subunit

    International Nuclear Information System (INIS)

    Bidart, J.M.; Troalen, F.; Salesse, R.; Bousfield, G.R.; Bohuon, C.J.; Bellet, D.H.

    1987-01-01

    We describe a first attempt to study the antibody-combining sites recognized by monoclonal antibodies raised against the beta-subunit of human choriogonadotropin (hCG). Two groups of antibodies were first defined by their ability to recognize only the free beta-subunit or the free and combined subunit. Antibodies FBT-11 and FBT-11-L bind only to hCG beta-subunit but not to hCG, whereas antibodies FBT-10 and D1E8 bind to both the beta-subunit and the hormone. In both cases, the antigenic determinants were localized to the core of the protein (residues 1-112), indicating the weak immunogenicity of the specific carboxyl-terminal extension of hCG-beta. Nine synthetic peptides spanning different regions of hCG-beta and lutropin-beta were assessed for their capacity to inhibit antibody binding. A synthetic peptide inclusive of the NH2-terminal region (residues 1-7) of the hCG beta-subunit was found to inhibit binding to the radiolabeled subunit of a monoclonal antibody specific for free hCG-beta (FBT-11). Further delineation of the antigenic site recognized by this antibody provided evidence for the involvement of fragment 82-92. Moreover, monoclonal antibody FBT-11 inhibited the recombination of hCG-beta to hCG-alpha, indicating that its antigenic determinant might be located nearby or in the hCG-beta portion interacting with the alpha-subunit. Binding of monoclonal antibody FBT-10, corresponding to the second antigenic determinant, was weakly inhibited by fragment 82-105 and did not impair the recombination of the hCG beta-subunit to the hCG alpha-subunit. Its combining site appeared to be located in a region of the intact native choriogonadotropin present at the surface of the hormone-receptor complex

  3. The nightmare of indeterminate follicular proliferations: when liquid-based cytology and ancillary techniques are not a moon landing but a realistic plan.

    Science.gov (United States)

    Rossi, Esther Diana; Fadda, Guido; Schmitt, Fernando

    2014-01-01

    Thyroid nodules are a common finding in the general population, including both nonneoplastic and neoplastic entities. Fine-needle aspiration cytology (FNAC) is the first tool for evaluating thyroid nodules. In spite of its high diagnostic accuracy, 25% of nodules result in the category of follicular neoplasms (FN), with varying risk of malignancy and different management strategies. The use of ancillary techniques is reshaping the practice of FNAC. These tools can significantly empower the morphological diagnosis and prognosis of thyroid nodules, allowing a more accurate prediction of the nature of the lesion. Several studies have underlined the role of single or multiple testing for the category of FN as strong indicators of cancer. Every cytological preparation can be used for the application of ancillary techniques but the introduction of liquid-based cytology (LBC) might facilitate the application. Our experience involving an immunocytochemical panel made up of HBME-1 and galectin-3 pointed to an 81% overall diagnostic accuracy in discriminating between low and high risk of malignancy in FN. The application of these techniques on LBC represents an adjunct to the morphological evaluation of FN. They represent a critical and challenging, but also a feasible, tool in the preoperative diagnoses, allowing specific prognostic and predictive details regardless of the cytological preparation. © 2014 S. Karger AG, Basel.

  4. 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.

  5. Compliance with technical codes becomes obligatory for receipt of feed-in tariff and ancillary services bonus for wind power plants in Germany

    Energy Technology Data Exchange (ETDEWEB)

    Boemer, Jens. C. [Ecofys Germany GmbH, Berlin (Germany). Power Systems and Markets Group; Delft Univ. of Technology (Netherlands). Electrical Power Systems Group; Burges, Karsten [Ecofys Germany GmbH, Berlin (Germany). Power Systems and Markets Group; Kumm, Thomas [VDE, Network Technology and Operation Forum (FNN), Berlin (Germany); Poeller, Markus [DIgSILENT GmbH, Gomaringen (Germany)

    2009-07-01

    The German government aims at a share of electricity generation based on renewable sources (RES-E) of more than 30% by 2020. A major part of this share will be reached by the use of wind power plants (WPP). Therefore, when amending the Renewable Energy Sources Act (EEG) in June 2008, the German government acknowledged the importance of enhanced technical requirements for WPP and announced the development of a respective technical ordinance. For all WPP that go into operation after June 2010, full grid code-compliance becomes a precondition for privileged network access, receipt of feed-in tariff, and extra payments (''ancillary services bonus''). The basis for the technical requirements in the EEG has been laid down by Medium-Voltage (MV)-Directive 2008 and the TransmissionCode 2007. However, the work of the authors - consulting the German government during the development of the technical ordinance - showed that the TransmissionCode 2007 needed careful review and some clarifying specifications were proposed. Eventually, the so called ''Ancillary Services Ordinance'' for wind power plants went into power in July 2009. The interrelation between all three grid codes as well as their specifications and implications for renewable energy sources generators, with special regard to wind power plants, are presented in this paper. (orig.)

  6. 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.

  7. Subunit association as the stabilizing determinant for archaeal methionine adenosyltransferases.

    Science.gov (United States)

    Garrido, Francisco; Alfonso, Carlos; Taylor, John C; Markham, George D; Pajares, María A

    2009-07-01

    Archaea contain a class of methionine adenosyltransferases (MATs) that exhibit substantially higher stability than their mesophilic counterparts. Their sequences are highly divergent, but preserve the essential active site motifs of the family. We have investigated the origin of this increased stability using chemical denaturation experiments on Methanococcus jannaschii MAT (Mj-MAT) and mutants containing single tryptophans in place of tyrosine residues. The results from fluorescence, circular dichroism, hydrodynamic, and enzyme activity measurements showed that the higher stability of Mj-MAT derives largely from a tighter association of its subunits in the dimer. Local fluorescence changes, interpreted using secondary structure predictions, further identify the least stable structural elements as the C-terminal ends of beta-strands E2 and E6, and the N-terminus of E3. Dimer dissociation however requires a wider perturbation of the molecule. Additional analysis was initially hindered by the lack of crystal structures for archaeal MATs, a limitation that we overcame by construction of a 3D-homology model of Mj-MAT. This model predicts preservation of the chain topology and three-domain organization typical of this family, locates the least stable structural elements at the flat contact surface between monomers, and shows that alterations in all three domains are required for dimer dissociation.

  8. 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.

  9. 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.

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

    DEFF Research Database (Denmark)

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

    2007-01-01

    Adenosine to inosine (A-to-I) pre-mRNA editing by the ADAR enzyme family has the potential to increase the variety of the proteome. This editing by adenosine deamination is essential in mammals for a functional brain. To detect novel substrates for A-to-I editing we have used an experimental method...... 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......, is a substrate for editing by both ADAR1 and ADAR2. Editing of the Gabra-3 mRNA recodes an isoleucine to a methionine. The extent of editing is low at birth but increases with age, reaching close to 100% in the adult brain. We therefore propose that editing of the Gabra-3 mRNA is important for normal brain...

  11. 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.

  12. 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.

  13. 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)

  14. Crystal structure of the P pilus rod subunit PapA.

    Directory of Open Access Journals (Sweden)

    Denis Verger

    2007-05-01

    Full Text Available P pili are important adhesive fibres involved in kidney infection by uropathogenic Escherichia coli strains. P pili are assembled by the conserved chaperone-usher pathway, which involves the PapD chaperone and the PapC usher. During pilus assembly, subunits are incorporated into the growing fiber via the donor-strand exchange (DSE mechanism, whereby the chaperone's G1 beta-strand that complements the incomplete immunoglobulin-fold of each subunit is displaced by the N-terminal extension (Nte of an incoming subunit. P pili comprise a helical rod, a tip fibrillum, and an adhesin at the distal end. PapA is the rod subunit and is assembled into a superhelical right-handed structure. Here, we have solved the structure of a ternary complex of PapD bound to PapA through donor-strand complementation, itself bound to another PapA subunit through DSE. This structure provides insight into the structural basis of the DSE reaction involving this important pilus subunit. Using gel filtration chromatography and electron microscopy on a number of PapA Nte mutants, we establish that PapA differs in its mode of assembly compared with other Pap subunits, involving a much larger Nte that encompasses not only the DSE region of the Nte but also the region N-terminal to it.

  15. Structural characterization of recombinant crustacyanin subunits from the lobster Homarus americanus

    International Nuclear Information System (INIS)

    Ferrari, Michele; Folli, Claudia; Pincolini, Elisa; McClintock, Timothy S.; Rössle, Manfred; Berni, Rodolfo; Cianci, Michele

    2012-01-01

    The two recombinant apo subunits H1 and H2 from H. americanus have been structurally characterized. Reconstitution studies with astaxanthin reproduced the bathochromic shift of 85–95 nm typical of the natural crustacyanin subunits. Crustacean crustacyanin proteins are linked to the production and modification of carapace colour, with direct implications for fitness and survival. Here, the structural and functional properties of the two recombinant crustacyanin subunits H 1 and H 2 from the American lobster Homarus americanus are reported. The two subunits are structurally highly similar to the corresponding natural apo crustacyanin CRTC and CRTA subunits from the European lobster H. gammarus. Reconstitution studies of the recombinant crustacyanin proteins H 1 and H 2 with astaxanthin reproduced the bathochromic shift of 85–95 nm typical of the natural crustacyanin subunits from H. gammarus in complex with astaxanthin. Moreover, correlations between the presence of crustacyanin genes in crustacean species and the resulting carapace colours with the spectral properties of the subunits in complex with astaxanthin confirmed this genotype–phenotype linkage

  16. Self-subunit swapping occurs in another gene type of cobalt nitrile hydratase.

    Directory of Open Access Journals (Sweden)

    Yi Liu

    Full Text Available 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.

  17. 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.

  18. 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

  19. 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.

  20. Effect of glutenin subunits on the baking quality of Brazilian wheat genotypes

    OpenAIRE

    Costa, Mariana Souza; Scholz, Maria Brígida dos Santos; Miranda, Martha Zavariz; Franco, Célia Maria Landi

    2017-01-01

    ABSTRACT This study aimed to evaluate the effect of the high and low molecular weight glutenin subunits on the grain traits of sixteen Brazilian wheat genotypes. Grain hardness index, milling traits, physicochemical and rheological properties of the flour, and specific volume and firmness of the bread were evaluated. Physicochemical properties of the flour were not influenced by glutenin subunits. Genotypes with subunits at the Glu-B1 (17+18 or 7+8), Glu-D1 (5+10), and Glu-A3 (b) were associa...

  1. Inhibition of herpesvirus and influenza virus replication by blocking polymerase subunit interactions.

    Science.gov (United States)

    Palù, Giorgio; Loregian, Arianna

    2013-09-01

    Protein-protein interactions (PPIs) play a key role in many biological processes, including virus replication in the host cell. Since most of the PPIs are functionally essential, a possible strategy to inhibit virus replication is based on the disruption of viral protein complexes by peptides or small molecules that interfere with subunit interactions. In particular, an attractive target for antiviral drugs is the binding between the subunits of essential viral enzymes. This review describes the development of new antiviral compounds that inhibit herpesvirus and influenza virus replication by blocking interactions between subunit proteins of their polymerase complexes. Copyright © 2013 Elsevier B.V. All rights reserved.

  2. Protein Kinase A Regulatory Subunits in Human Adipose Tissue

    Science.gov (United States)

    Mantovani, Giovanna; Bondioni, Sara; Alberti, Luisella; Gilardini, Luisa; Invitti, Cecilia; Corbetta, Sabrina; Zappa, Marco A.; Ferrero, Stefano; Lania, Andrea G.; Bosari, Silvano; Beck-Peccoz, Paolo; Spada, Anna

    2009-01-01

    OBJECTIVE—In human adipocytes, the cAMP-dependent pathway mediates signals originating from β-adrenergic activation, thus playing a key role in the regulation of important metabolic processes, i.e., lipolysis and thermogenesis. Cyclic AMP effects are mainly mediated by protein kinase A (PKA), whose R2B regulatory isoform is the most expressed in mouse adipose tissue, where it protects against diet-induced obesity and fatty liver development. The aim of the study was to investigate possible differences in R2B expression, PKA activity, and lipolysis in adipose tissues from obese and nonobese subjects. RESEARCH DESIGN AND METHODS—The expression of the different PKA regulatory subunits was evaluated by immunohistochemistry, Western blot, and real-time PCR in subcutaneous and visceral adipose tissue samples from 20 nonobese and 67 obese patients. PKA activity and glycerol release were evaluated in total protein extract and adipocytes isolated from fresh tissue samples, respectively. RESULTS—Expression techniques showed that R2B was the most abundant regulatory protein, both at mRNA and protein level. Interestingly, R2B mRNA levels were significantly lower in both subcutaneous and visceral adipose tissues from obese than nonobese patients and negatively correlated with BMI, waist circumference, insulin levels, and homeostasis model assessment of insulin resistance. Moreover, both basal and stimulated PKA activity and glycerol release were significantly lower in visceral adipose tissue from obese patients then nonobese subjects. CONCLUSIONS—Our results first indicate that, in human adipose tissue, there are important BMI-related differences in R2B expression and PKA activation, which might be included among the multiple determinants involved in the different lipolytic response to β-adrenergic activation in obesity. PMID:19095761

  3. P. berghei telomerase subunit TERT is essential for parasite survival.

    Directory of Open Access Journals (Sweden)

    Agnieszka A Religa

    Full Text Available Telomeres define the ends of chromosomes protecting eukaryotic cells from chromosome instability and eventual cell death. The complex regulation of telomeres involves various proteins including telomerase, which is a specialized ribonucleoprotein responsible for telomere maintenance. Telomeres of chromosomes of malaria parasites are kept at a constant length during blood stage proliferation. The 7-bp telomere repeat sequence is universal across different Plasmodium species (GGGTTT/CA, though the average telomere length varies. The catalytic subunit of telomerase, telomerase reverse transcriptase (TERT, is present in all sequenced Plasmodium species and is approximately three times larger than other eukaryotic TERTs. The Plasmodium RNA component of TERT has recently been identified in silico. A strategy to delete the gene encoding TERT via double cross-over (DXO homologous recombination was undertaken to study the telomerase function in P. berghei. Expression of both TERT and the RNA component (TR in P. berghei blood stages was analysed by Western blotting and Northern analysis. Average telomere length was measured in several Plasmodium species using Telomere Restriction Fragment (TRF analysis. TERT and TR were detected in blood stages and an average telomere length of ∼ 950 bp established. Deletion of the tert gene was performed using standard transfection methodologies and we show the presence of tert- mutants in the transfected parasite populations. Cloning of tert- mutants has been attempted multiple times without success. Thorough analysis of the transfected parasite populations and the parasite obtained from extensive parasite cloning from these populations provide evidence for a so called delayed death phenotype as observed in different organisms lacking TERT. The findings indicate that TERT is essential for P. berghei cell survival. The study extends our current knowledge on telomere biology in malaria parasites and validates further

  4. 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

  5. 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...

  6. 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

    Although Chou-Fasman calculations of the secondary structure of recombinant casein kinase 2 subunits alpha and beta suggest they have a similar overall conformation, circular dichroism (CD) studies show that substantial differences in the conformation of the two subunits exist. In addition......, 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...

  7. Stereocontrolled Synthesis of the C(1)-C(11) Subunit of the Iejimalides

    DEFF Research Database (Denmark)

    Mendlik, Matthew T.; Cottard, Muriel; Rein, Tobias

    1997-01-01

    An enantioselective synthesis of the C(1)-C(11) subunit of the iejimalides has been accomplished through a combination of an asymmetric Homer-Wadsworth-Emmons condensation and a chiral pool approach. (C) 1997 Elsevier Science Ltd....

  8. Identification of a conserved archaeal RNA polymerase subunit contacted by the basal transcription factor TFB.

    Science.gov (United States)

    Magill, C P; Jackson, S P; Bell, S D

    2001-12-14

    Archaea possess two general transcription factors that are required to recruit RNA polymerase (RNAP) to promoters in vitro. These are TBP, the TATA-box-binding protein and TFB, the archaeal homologue of TFIIB. Thus, the archaeal and eucaryal transcription machineries are fundamentally related. In both RNAP II and archaeal transcription systems, direct contacts between TFB/TFIIB and the RNAP have been demonstrated to mediate recruitment of the polymerase to the promoter. However the subunit(s) directly contacted by these factors has not been identified. Using systematic yeast two-hybrid and biochemical analyses we have identified an interaction between the N-terminal domain of TFB and an evolutionarily conserved subunit of the RNA polymerase, RpoK. Intriguingly, homologues of RpoK are found in all three nuclear RNA polymerases (Rpb6) and also in the bacterial RNA polymerase (omega-subunit).

  9. Subunit architecture and functional modular rearrangements of the transcriptional mediator complex.

    Science.gov (United States)

    Tsai, Kuang-Lei; Tomomori-Sato, Chieri; Sato, Shigeo; Conaway, Ronald C; Conaway, Joan W; Asturias, Francisco J

    2014-06-05

    The multisubunit Mediator, comprising ∼30 distinct proteins, plays an essential role in gene expression regulation by acting as a bridge between DNA-binding transcription factors and the RNA polymerase II (RNAPII) transcription machinery. Efforts to uncover the Mediator mechanism have been hindered by a poor understanding of its structure, subunit organization, and conformational rearrangements. By overcoming biochemical and image analysis hurdles, we obtained accurate EM structures of yeast and human Mediators. Subunit localization experiments, docking of partial X-ray structures, and biochemical analyses resulted in comprehensive mapping of yeast Mediator subunits and a complete reinterpretation of our previous Mediator organization model. Large-scale Mediator rearrangements depend on changes at the interfaces between previously described Mediator modules, which appear to be facilitated by factors conducive to transcription initiation. Conservation across eukaryotes of Mediator structure, subunit organization, and RNA polymerase II interaction suggest conservation of fundamental aspects of the Mediator mechanism. Copyright © 2014 Elsevier Inc. All rights reserved.

  10. 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.

  11. Two subunits of human ORC are dispensable for DNA replication and proliferation.

    Science.gov (United States)

    Shibata, Etsuko; Kiran, Manjari; Shibata, Yoshiyuki; Singh, Samarendra; Kiran, Shashi; Dutta, Anindya

    2016-12-01

    The six-subunit Origin Recognition Complex (ORC) is believed to be an essential eukaryotic ATPase that binds to origins of replication as a ring-shaped heterohexamer to load MCM2-7 and initiate DNA replication. We have discovered that human cell lines in culture proliferate with intact chromosomal origins of replication after disruption of both alleles of ORC2 or of the ATPase subunit, ORC1 . The ORC1 or ORC2 -depleted cells replicate with decreased chromatin loading of MCM2-7 and become critically dependent on another ATPase, CDC6, for survival and DNA replication. Thus, either the ORC ring lacking a subunit, even its ATPase subunit, can load enough MCM2-7 in partnership with CDC6 to initiate DNA replication, or cells have an ORC-independent, CDC6-dependent mechanism to load MCM2-7 on origins of replication.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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

  17. Pharmacological consequences of the coexpression of BK channel α and auxiliary β subunits

    Directory of Open Access Journals (Sweden)

    Yolima P. Torres

    2014-10-01

    Full Text Available 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 Ca+2-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 & 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 Ca+2 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.

  18. 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 Ca(2+) concentration, the large conductance voltage- and Ca(2+)-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 Ca(2+)-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 Ca(2+) 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.

  19. 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

  20. Increased Expression of Laminin Subunit Alpha 1 Chain by dCas9-VP160

    OpenAIRE

    Perrin, Arnaud; Rousseau, Jo?l; Tremblay, Jacques P.

    2016-01-01

    Laminin-111 protein complex links the extracellular matrix to integrin α7β1 in sarcolemma, thus replacing in dystrophic muscles links normally insured by the dystrophin complex. Laminin-111 injection in mdx mouse stabilized sarcolemma, restored serum creatine kinase to wild-type levels, and protected muscles from exercised-induced damages. These results suggested that increased laminin-111 is a potential therapy for DMD. Laminin subunit beta 1 and laminin subunit gamma 1 are expressed in adul...

  1. Neutron scattering and the 30 S ribosomal subunit of E. coli

    International Nuclear Information System (INIS)

    Moore, P.B.; Engelman, D.M.; Langer, J.A.; Ramakrishnan, V.R.; Schindler, D.G.; Schoenborn, B.P.; Sillers, I.Y.; Yabuki, S.

    1982-01-01

    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. 30 references, 5 figures

  2. Distribution of AMPA-type glutamate receptor subunits in the chick visual system

    Directory of Open Access Journals (Sweden)

    Pires R.S.

    1997-01-01

    Full Text Available Several glutamate receptor (GluR subunits have been characterized during the past few years. In the present study, subunit-specific antisera were used to determine the distribution of the AMPA-type glutamate receptor subunits GluR1-4 in retinorecipient areas of the chick brain. Six white leghorn chicks (Gallus gallus, 7-15 days old, unknown sex were deeply anesthetized and perfused with 4% buffered paraformaldehyde and brain sections were stained using immunoperoxidase techniques. The AMPA-type glutamate receptor subunits GluR1, GluR2/3 and GluR4 were present in several retinorecipient areas, with varying degrees of colocalization. For example, perikarya in layers 2, 3, and 5 of the optic tectum contained GluR1, whereas GluR2/3 subunits appeared mainly in neurons of layer 13. The GluR4 subunit was only detected in a few cells of the tectal layer 13. GluR1 and GluR2/3 were observed in neurons of the nucleus geniculatus lateralis ventralis, whereas GluR4 was only present in its neuropil. Somata in the accessory optic nucleus appeared to contain GluR2/3 and GluR4, whereas GluR1 was the dominant subunit in the neuropil of this nucleus. These results suggest that different subpopulations of visual neurons might express different combinations of AMPA-type GluR subunits, which in turn might generate different synaptic responses to glutamate derived from retinal ganglion cell axons

  3. Crystal structure of Agaricus bisporus mushroom tyrosinase: identity of the tetramer subunits and interaction with tropolone.

    Science.gov (United States)

    Ismaya, Wangsa T; Rozeboom, Henriëtte J; Weijn, Amrah; Mes, Jurriaan J; Fusetti, Fabrizia; Wichers, Harry J; Dijkstra, Bauke W

    2011-06-21

    Tyrosinase catalyzes the conversion of phenolic compounds into their quinone derivatives, which are precursors for the formation of melanin, a ubiquitous pigment in living organisms. Because of its importance for browning reactions in the food industry, the tyrosinase from the mushroom Agaricus bisporus has been investigated in depth. In previous studies the tyrosinase enzyme complex was shown to be a H(2)L(2) tetramer, but no clues were obtained of the identities of the subunits, their mode of association, and the 3D structure of the complex. Here we unravel this tetramer at the molecular level. Its 2.3 Å resolution crystal structure is the first structure of the full fungal tyrosinase complex. The complex comprises two H subunits of ∼392 residues and two L subunits of ∼150 residues. The H subunit originates from the ppo3 gene and has a fold similar to other tyrosinases, but it is ∼100 residues larger. The L subunit appeared to be the product of orf239342 and has a lectin-like fold. The H subunit contains a binuclear copper-binding site in the deoxy-state, in which three histidine residues coordinate each copper ion. The side chains of these histidines have their orientation fixed by hydrogen bonds or, in the case of His85, by a thioether bridge with the side chain of Cys83. The specific tyrosinase inhibitor tropolone forms a pre-Michaelis complex with the enzyme. It binds near the binuclear copper site without directly coordinating the copper ions. The function of the ORF239342 subunits is not known. Carbohydrate binding sites identified in other lectins are not conserved in ORF239342, and the subunits are over 25 Å away from the active site, making a role in activity unlikely. The structures explain how calcium ions stabilize the tetrameric state of the enzyme.

  4. 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

  5. Proteasome (Prosome Subunit Variations during the Differentiation of Myeloid U937 Cells

    Directory of Open Access Journals (Sweden)

    Laurent Henry

    1997-01-01

    Full Text Available 20S proteasomes (prosomes/multicatalytic proteinase are protein particles built of 28 subunits in variable composition. We studied the changes in proteasome subunit composition during the differentiation of U937 cells induced by phorbol‐myristate‐acetate or retinoic acid plus 1,25‐dihydroxy‐cholecalciferol by western blot, flow cytometry and immuno‐fluorescence. p25K (C3, p27K (IOTA and p30/33K (C2 subunits were detected in both the nucleus and cytoplasm of undifferentiated cells. Flow cytometry demonstrated a biphasic decrease in proteasome subunits detection during differentiation induced by RA+VD. PMA caused an early transient decrease in these subunits followed by a return to their control level, except for p30/33K, which remained low. Immuno‐fluorescence also showed differences in the cytolocalization of the subunits, with a particular decrease in antigen labeling in the nucleus of RA+VD‐induced cells, and a scattering in the cytoplasm and a reorganization in the nucleus of PMA‐induced cells. Small amounts of proteasomal proteins were seen on the outer membrane of non‐induced cells; these membrane proteins disappeared when treated with RA+VD, whereas some increased on PMA‐induced cells. The differential changes in the distribution and type of proteasomes in RA+VD and PMA‐induced cells indicate that, possibly, 20S proteasomes may play a role in relation to the mechanisms of differentiation and the inducer used.

  6. Characterization and application of a radioimmunoassay for reduced, carboxymethylated human luteinizing hormone α-subunit

    International Nuclear Information System (INIS)

    Keutmann, H.T.; Beitins, I.Z.; Johnson, L.; McArthur, J.W.

    1978-01-01

    We have established a double antibody RIA using a rabbit antiserum prepared against reduced, carboxymethylated (RCXM) human LH α-subunit, with RCXM-α as tracer and standard. This antiserum did not cross-react with any native gonadotropins or subunit, and reacted only weakly with RCXM-α. A tryptic digest of RCXM α-subunit was completely reactive, while chymotryptic digestion abolished all immunoreactivity. By testing with separate tryptic fragments, the recognition site could be localized to a segment close to the amino-terminus of the peptide chain. When applied to measurement of serum and urine, an immunoreactive species, parallel to RCXM α-subunit by serial dilution, was found in concentrations of 1-2 ng/ml in serum and 3-4 ng/ml in urine. Similar levels of the immunoreactive component were found in conditions of elevated gonadotropins (e.g. pregnancy) as well as gonadotropin deficiency (panhypopituitarism and Kallmann's syndrome). After stimulation with LHRH, no rise was noted at times up to 6 h despite the fact that both LH and LH-α were elevated. The data indicate that the sequence-specific antiserum may be detecting an immunoreactive form of α-subunit of LH whose kinetics of appearance and disappearance differs from those of the native subunit

  7. 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.

  8. Effect of glutenin subunits on the baking quality of Brazilian wheat genotypes

    Directory of Open Access Journals (Sweden)

    Mariana Souza Costa

    Full Text Available ABSTRACT This study aimed to evaluate the effect of the high and low molecular weight glutenin subunits on the grain traits of sixteen Brazilian wheat genotypes. Grain hardness index, milling traits, physicochemical and rheological properties of the flour, and specific volume and firmness of the bread were evaluated. Physicochemical properties of the flour were not influenced by glutenin subunits. Genotypes with subunits at the Glu-B1 (17+18 or 7+8, Glu-D1 (5+10, and Glu-A3 (b were associated with strong flours and bread with high specific volume and low firmness. The subunits at the Glu-A1 and Glu-B3 had no effect on the rheological properties of the dough and bread quality, while the subunit 2+12 at Glu-D1 negatively affected the resistance to extension, and specific volume and firmness of the bread. Specific volume and firmness of the bread were influenced by the rheological properties of the dough, while the flour protein content was not important to define wheat quality. The identification of glutenin subunits at different loci along with the rheological tests of the flour are fundamental in estimating the potential use of different materials developed in wheat breeding.

  9. 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.

  10. Pemanfaatan Limbah Kulit Jagung untuk Produk Modular dengan Teknik Pilin

    Directory of Open Access Journals (Sweden)

    Artarita Ginting

    2016-04-01

    Full Text Available ABSTRAKPengembangan industri kreatif tanpa limbah merupakan persyaratan penting bagi keseimbangan yang baik antara peningkatan usaha industri, daya dukung alam dan kesejahteraan manusia yang tinggal di lingkungan tersebut. Sangat sedikit usaha para pelaku industri untuk mengoptimalkan manfaat sebuah hasil alam hingga tidak menghasilkan limbah sama sekali. Kulit jagung merupakan salah satu limbah rumah tangga dan industri kecil yang jumlahnya berlimpah namun kurang optimal dalam pemanfaatannya. Produksi dan konsumsi jagung merupakan bagian dari satu sistem kehidupan yang utuh sehingga patut dipertimbangkan strategi pelaksanaannya agar daya dukung lingkungan tetap kuat. Penelitian eksperimental bahan kulit jagung ini bertujuan memanfaatkan limbah kulit jagung sebagai bahan alternatif produk kerajinan secara optimal tanpa menghasilkan limbah kembali. Dalam penelitian yang menggunakan metode penelitian eksperimen bahan posttest-only, diperoleh kesimpulan bahwa pengawetan dengan rendaman CH3COOH selama 24 jam dan proses penjemuran selama 3 jam pada jam 9 pagi hingga jam 12 siang menghasilkan serat kulit jagung yang memiliki kekuatan tarik paling tinggi dan warna yang cerah. Sedangkan teknik pemilinan membantu untuk menambah kekuatan tarik melalui kepadatan dari hasil pilinan kulit jagung. Hasil penelitian eksperimen bahan digunakan untuk membuat spesifikasi performa produk bagi konsep perancangan desain rak anyam modular yang diwujudkan dengan teknik sambung pasak yang praktis dalam penggunaannya. Kata kunci: tanpa limbah, pilinan kulit jagung, kaleng bekas, modular ABSTRACT  The development of zero waste creative industry is a vital prerequisite for a healthy balance between industrial development, nature support capacity and community welfare within the area. There are only a few number of industries that consider to optimalized their raw materials towards zero waste goal. Corn husk is one of the industrial and residential waste that is under utilize although available in abundance. Production and consumption is a part of the whole living system therefore both should be parts of the industrial consideration in asssuring nature support capability. This posttest-only experimental research of corn husk is intended to optimally utilize corn husk as a raw material alternative in craft industry towards zero waste production. The highest tensile force and the brightest color is attained by soaking corn husk in CH3COOH chemical liquid and drying them for 3 hours from 9.00 AM to 12.00PM. The twisting technic has enhanced its tensile capability through its twisting density. The result of the material experimental research is used to create a product performance specification for designing a modular woven rack, produced by using a practical rod pegs connection. Keywords: zero waste, twisted corn husk, used tin can, modular

  11. Discovering the bacterial circular proteins : bacteriocins, cyanobactins, and pilins

    NARCIS (Netherlands)

    Montalban-Lopez, Manuel; Sanchez-Hidalgo, Marina; Cebrian, Ruben; Maqueda, Mercedes

    2012-01-01

    Over recent years, several examples of natural ribosomally synthesized circular proteins and peptides from diverse organisms have been described. They are a group of proteins for which the precursors must be post-translationally modified to join the N and C termini with a peptide bond. This feature

  12. Reactivation of the chloroplast CF1-ATPase beta subunit by trace amounts of the CF1 alpha subunit suggests a chaperonin-like activity for CF1 alpha.

    Science.gov (United States)

    Avni, A; Avital, S; Gromet-Elhanan, Z

    1991-04-25

    Incubation of tobacco and lettuce thylakoids with 2 M LiCl in the presence of MgATP removes the beta subunit from their CF1-ATPase (CF1 beta) together with varying amounts of the CF1 alpha subunit (CF1 alpha). These 2 M LiCl extracts, as with the one obtained from spinach thylakoids (Avital, S., and Gromet-Elhanan, Z. (1991) J. Biol. Chem. 266, 7067-7072), could form active hybrid ATPases when reconstituted into inactive beta-less Rhodospirillum rubrum chromatophores. Pure CF1 beta fractions that have been isolated from these extracts could not form such active hybrids by themselves, but could do so when supplemented with trace amounts (less than 5%) of CF1 alpha. A mitochondrial F1-ATPase alpha subunit was recently reported to be a heat-shock protein, having two amino acid sequences that show a highly conserved identity with sequences found in molecular chaperones (Luis, A. M., Alconada, A., and Cuezva, J. M. (1990) J. Biol. Chem. 265, 7713-7716). These sequences are also conserved in CF1 alpha isolated from various plants, but not in F1 beta subunits. The above described reactivation of CF1 beta by trace amounts of CF1 alpha could thus be due to a chaperonin-like function of CF1 alpha, which involves the correct, active folding of isolated pure CF1 beta.

  13. 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

  14. 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.

  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. 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.

  17. 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.

  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. 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

  20. 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.

  1. 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.

  2. 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

  3. 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.

  4. 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.

  5. Assessing subunit dependency of the Plasmodium proteasome using small molecule inhibitors and active site probes.

    Science.gov (United States)

    Li, Hao; van der Linden, Wouter A; Verdoes, Martijn; Florea, Bogdan I; McAllister, Fiona E; Govindaswamy, Kavitha; Elias, Joshua E; Bhanot, Purnima; Overkleeft, Herman S; Bogyo, Matthew

    2014-08-15

    The ubiquitin-proteasome system (UPS) is a potential pathway for therapeutic intervention for pathogens such as Plasmodium, the causative agent of malaria. However, due to the essential nature of this proteolytic pathway, proteasome inhibitors must avoid inhibition of the host enzyme complex to prevent toxic side effects. The Plasmodium proteasome is poorly characterized, making rational design of inhibitors that induce selective parasite killing difficult. In this study, we developed a chemical probe that labels all catalytic sites of the Plasmodium proteasome. Using this probe, we identified several subunit selective small molecule inhibitors of the parasite enzyme complex. Treatment with an inhibitor that is specific for the β5 subunit during blood stage schizogony led to a dramatic decrease in parasite replication while short-term inhibition of the β2 subunit did not affect viability. Interestingly, coinhibition of both the β2 and β5 catalytic subunits resulted in enhanced parasite killing at all stages of the blood stage life cycle and reduced parasite levels in vivo to barely detectable levels. Parasite killing was achieved with overall low host toxicity, something that has not been possible with existing proteasome inhibitors. Our results highlight differences in the subunit dependency of the parasite and human proteasome, thus providing a strategy for development of potent antimalarial drugs with overall low host toxicity.

  6. G-protein α-subunit expression, myristoylation, and membrane association in COS cells

    International Nuclear Information System (INIS)

    Mumby, S.M.; Gilman, A.G.; Heukeroth, R.O.; Gordon, J.I.

    1990-01-01

    Myristolyation of seven different α subunits of guanine nucleotide-binding regulatory proteins (G proteins) was examined by expressing these proteins in monkey kidney COS cells. Metabolic labeling studies of cells transfected with cytomegalovirus-based expression vectors indicated that [ 3 H]myristate was incorporated into α i1 , α i2 , α i3 , α 0 , and α 1 , and α z but not α s subunits. The role of myristoylation in the association of α subunits with membranes was analyzed by site-directed mutagenesis and by substitution of myristate with a less hydrophobic analog, 10-(propoxy)decanoate (11-oxamyristate). Myristoylation of α 0 was blocked when an alanine residue was substituted for its amino-terminal glycine, as was association of the protein with membranes. Substitution of the myristoyl group with 11-oxamyristate affected the cellular distribution of a subset of acylated α subunits. The results are consistent with a model wherein the hydrophobic interaction of myristate with the bilayer permits continued association of the protein with the plasma membrane when G-protein α subunits dissociated from βγ

  7. Highly diverged novel subunit composition of apicomplexan F-type ATP synthase identified from Toxoplasma gondii

    KAUST Repository

    Salunke, Rahul

    2018-05-14

    The mitochondrial F-type ATP synthase, a multi-subunit nanomotor, is critical for maintaining cellular ATP levels. In Toxoplasma gondii and other apicomplexan parasites, many subunit components, necessary for proper assembly and functioning of this enzyme, appear to be missing. Here, we report the identification of 20 novel subunits of T. gondii F-type ATP synthase from mass spectrometry analysis of partially purified monomer (~600 kDa) and dimer (>1 MDa) forms of the enzyme. Despite extreme sequence diversification, key FO subunits, a, b and d, can be identified from conserved structural features. Orthologs for these proteins are restricted to apicomplexan, chromerid and dinoflagellate species. Interestingly, their absence in ciliates indicates a major diversion, with respect to subunit composition of this enzyme, within the alveolate clade. Discovery of these highly diversified novel components of the apicomplexan F-type ATP synthase complex will facilitate the development of novel anti-parasitic agents. Structural and functional characterization of this unusual enzyme complex will advance our fundamental understanding of energy metabolism in apicomplexan species.

  8. Highly diverged novel subunit composition of apicomplexan F-type ATP synthase identified from Toxoplasma gondii

    KAUST Repository

    Salunke, Rahul; Mourier, Tobias; Banerjee, Manidipa; Pain, Arnab; Shanmugam, Dhanasekaran

    2018-01-01

    The mitochondrial F-type ATP synthase, a multi-subunit nanomotor, is critical for maintaining cellular ATP levels. In Toxoplasma gondii and other apicomplexan parasites, many subunit components, necessary for proper assembly and functioning of this enzyme, appear to be missing. Here, we report the identification of 20 novel subunits of T. gondii F-type ATP synthase from mass spectrometry analysis of partially purified monomer (~600 kDa) and dimer (>1 MDa) forms of the enzyme. Despite extreme sequence diversification, key FO subunits, a, b and d, can be identified from conserved structural features. Orthologs for these proteins are restricted to apicomplexan, chromerid and dinoflagellate species. Interestingly, their absence in ciliates indicates a major diversion, with respect to subunit composition of this enzyme, within the alveolate clade. Discovery of these highly diversified novel components of the apicomplexan F-type ATP synthase complex will facilitate the development of novel anti-parasitic agents. Structural and functional characterization of this unusual enzyme complex will advance our fundamental understanding of energy metabolism in apicomplexan species.

  9. 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.

  10. 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.

  11. 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.

  12. 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

  13. 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...

  14. Isolation and characterization of a monoclonal anti CK-2 alpha subunit antibody of the IgG1 subclass

    DEFF Research Database (Denmark)

    Schmidt-Spaniol, I; Boldyreff, B; Issinger, O G

    1992-01-01

    A monoclonal antibody was produced against the recombinant human alpha subunit of CK-2. The antibody was of the IgG1 subclass and it was isolated from serum-free cell culture media and purified by affinity chromatography on Protein G Sepharose. The antibody can be used to detect specifically the CK......-2 alpha subunit in immunoblots from tissue extracts. An ELISA detection test was also established which also allows the identification of the CK-2 alpha subunit....

  15. MPC1-like Is a Placental Mammal-specific Mitochondrial Pyruvate Carrier Subunit Expressed in Postmeiotic Male Germ Cells

    OpenAIRE

    Vanderperre, Benoît; Cermakova, Kristina; Escoffier Breancon, Jessica; Kaba, Mayis; Bender, Tom; Nef, Serge; Martinou, Jean-Claude

    2016-01-01

    Selective transport of pyruvate across the inner mitochondrial membrane by the mitochondrial pyruvate carrier (MPC) is a fundamental step that couples cytosolic and mitochondrial metabolism. The recent molecular identification of the MPC complex has revealed two interacting subunits, MPC1 and MPC2. Although in yeast, an additional subunit, MPC3, can functionally replace MPC2, no alternative MPC subunits have been described in higher eukaryotes. Here, we report for the first time the existence...

  16. 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...

  17. Suppression of 19S proteasome subunits marks emergence of an altered cell state in diverse cancers.

    Science.gov (United States)

    Tsvetkov, Peter; Sokol, Ethan; Jin, Dexter; Brune, Zarina; Thiru, Prathapan; Ghandi, Mahmoud; Garraway, Levi A; Gupta, Piyush B; Santagata, Sandro; Whitesell, Luke; Lindquist, Susan

    2017-01-10

    The use of proteasome inhibitors to target cancer's dependence on altered protein homeostasis has been greatly limited by intrinsic and acquired resistance. Analyzing data from thousands of cancer lines and tumors, we find that those with suppressed expression of one or more 19S proteasome subunits show intrinsic proteasome inhibitor resistance. Moreover, such proteasome subunit suppression is associated with poor outcome in myeloma patients, where proteasome inhibitors are a mainstay of treatment. Beyond conferring resistance to proteasome inhibitors, proteasome subunit suppression also serves as a sentinel of a more global remodeling of the transcriptome. This remodeling produces a distinct gene signature and new vulnerabilities to the proapoptotic drug, ABT-263. This frequent, naturally arising imbalance in 19S regulatory complex composition is achieved through a variety of mechanisms, including DNA methylation, and marks the emergence of a heritably altered and therapeutically relevant state in diverse cancers.

  18. Cloning and Expression of Luteinizing Hormone Subunits in Chinese Hamster Ovary Cell Line

    Directory of Open Access Journals (Sweden)

    Zeinab Soleimanifar

    2016-10-01

    Full Text Available Background: Luteinizing hormone (LH was secreted by the stimulating cells of the testes and ovaries in the anterior pituitary gland. The application of this hormone is in the treatment of men and women with infertility and amenorrhea respectively.Materials and Methods: In the present study the alpha and beta subunits of human LH gene were cloned into the pEGFP-N1 expression vector and produced the recombinant LH hormone in Chinese hamster ovary (CHO eukaryotic system.Results: Alpha and beta subunits of LH hormone were cloned between NheI and BamHI cut sites of pEGFP_N1 expression plasmid and confirmed by PCR.  Hormone expression was evaluated in CHO cell line by Western blotting using the specific antibody.Conclusion: Alpha and beta subunits of LH hormone were expressed in CHO cell line perfectly.

  19. Progress in the development of subunit vaccines for gastrointestinal nematodes of ruminants.

    Science.gov (United States)

    Matthews, J B; Geldhof, P; Tzelos, T; Claerebout, E

    2016-12-01

    The global increase in anthelmintic resistant nematodes of ruminants, together with consumer concerns about chemicals in food, necessitates the development of alternative methods of control for these pathogens. Subunit recombinant vaccines are ideally placed to fill this gap. Indeed, they are probably the only valid option for the long-term control of ruminant parasitic nematodes given the increasing ubiquity of multidrug resistance in a range of worm species across the world. The development of a subunit multicellular parasite vaccine to the point of practical application would be a groundbreaking step in the control of these important endemic infections of livestock. This review summarizes the current status of subunit vaccine development for a number of important gastrointestinal nematodes of cattle and sheep, with a focus on the limitations and problems encountered thus far, and suggestions as to how these hurdles might be overcome. © 2016 John Wiley & Sons Ltd.

  20. Subunit Organisation of In Vitro Reconstituted HOPS and CORVET Multisubunit Membrane Tethering Complexes

    Science.gov (United States)

    Guo, Zhong; Johnston, Wayne; Kovtun, Oleksiy; Mureev, Sergey; Bröcker, Cornelia; Ungermann, Christian; Alexandrov, Kirill

    2013-01-01

    Biochemical and structural analysis of macromolecular protein assemblies remains challenging due to technical difficulties in recombinant expression, engineering and reconstitution of multisubunit complexes. Here we use a recently developed cell-free protein expression system based on the protozoan Leishmania tarentolae to produce in vitro all six subunits of the 600 kDa HOPS and CORVET membrane tethering complexes. We demonstrate that both subcomplexes and the entire HOPS complex can be reconstituted in vitro resulting in a comprehensive subunit interaction map. To our knowledge this is the largest eukaryotic protein complex in vitro reconstituted to date. Using the truncation and interaction analysis, we demonstrate that the complex is assembled through short hydrophobic sequences located in the C-terminus of the individual Vps subunits. Based on this data we propose a model of the HOPS and CORVET complex assembly that reconciles the available biochemical and structural data. PMID:24312556

  1. Gene expression patterns of oxidative phosphorylation complex I subunits are organized in clusters.

    Directory of Open Access Journals (Sweden)

    Yael Garbian

    Full Text Available After the radiation of eukaryotes, the NUO operon, controlling the transcription of the NADH dehydrogenase complex of the oxidative phosphorylation system (OXPHOS complex I, was broken down and genes encoding this protein complex were dispersed across the nuclear genome. Seven genes, however, were retained in the genome of the mitochondrion, the ancient symbiote of eukaryotes. This division, in combination with the three-fold increase in subunit number from bacteria (N = approximately 14 to man (N = 45, renders the transcription regulation of OXPHOS complex I a challenge. Recently bioinformatics analysis of the promoter regions of all OXPHOS genes in mammals supported patterns of co-regulation, suggesting that natural selection favored a mechanism facilitating the transcriptional regulatory control of genes encoding subunits of these large protein complexes. Here, using real time PCR of mitochondrial (mtDNA- and nuclear DNA (nDNA-encoded transcripts in a panel of 13 different human tissues, we show that the expression pattern of OXPHOS complex I genes is regulated in several clusters. Firstly, all mtDNA-encoded complex I subunits (N = 7 share a similar expression pattern, distinct from all tested nDNA-encoded subunits (N = 10. Secondly, two sub-clusters of nDNA-encoded transcripts with significantly different expression patterns were observed. Thirdly, the expression patterns of two nDNA-encoded genes, NDUFA4 and NDUFA5, notably diverged from the rest of the nDNA-encoded subunits, suggesting a certain degree of tissue specificity. Finally, the expression pattern of the mtDNA-encoded ND4L gene diverged from the rest of the tested mtDNA-encoded transcripts that are regulated by the same promoter, consistent with post-transcriptional regulation. These findings suggest, for the first time, that the regulation of complex I subunits expression in humans is complex rather than reflecting global co-regulation.

  2. Differential expression of AMPA-type glutamate receptor subunits during development of the chick optic tectum

    Directory of Open Access Journals (Sweden)

    Batista S.S.

    2002-01-01

    Full Text Available Glutamate receptors have been often associated with developmental processes. We used immunohistochemical techniques to evaluate the expression of the AMPA-type glutamate receptor (GluR subunits in the chick optic tectum (TeO. Chick embryos from the 5th through the 20th embryonic day (E5-E20 and one-day-old (P1 chicks were used. The three types of immunoreactivity evaluated (GluR1, GluR2/3, and GluR4 had different temporal and spatial expression patterns in the several layers of the TeO. The GluR1 subunit first appeared as moderate staining on E7 and then increased on E9. The mature GluR1 pattern included intense staining only in layer 5 of the TeO. The GluR2/3 subunits presented low expression on E5, which became intense on E7. The staining for GluR2/3 changed to very intense on E14 in tectal layer 13. Staining of layer 13 neurons is the most prominent feature of GluR immunoreactivity in the adult TeO. The GluR4 subunit generally presented the lowest expression starting on E7, which was similar to the adult pattern. Some instances of transient expression of GluR subunits were observed in specific cell populations from E9 through E20. These results demonstrate a differential expression of the GluR subunits in the embryonic TeO, adding information about their possible functions in the developmental processes of the visual system.

  3. Expression Profile of the Integrin Receptor Subunits in the Guinea Pig Sclera.

    Science.gov (United States)

    Wang, Kevin K; Metlapally, Ravikanth; Wildsoet, Christine F

    2017-06-01

    The ocular dimensional changes in myopia reflect increased scleral remodeling, and in high myopia, loss of scleral integrity leads to biomechanical weakening and continued scleral creep. As integrins, a type of cell surface receptors, have been linked to scleral remodeling, they represent potential targets for myopia therapies. As a first step, this study aimed to characterize the integrin subunits at the messenger RNA level in the sclera of the guinea pig, a more recently added but increasingly used animal model for myopia research. Primers for α and β integrin subunits were designed using NCBI/UCSC Genome Browser and Primer3 software tools. Total RNA was extracted from normal scleral tissue and isolated cultured scleral fibroblasts, as well as liver and lung, as reference tissues, all from guinea pig. cDNA was produced by reverse transcription, PCR was used to amplify products of predetermined sizes, and products were sequenced using standard methods. Guinea pig scleral tissue expressed all known integrin alpha subunits except αD and αE. The latter integrin subunits were also not expressed by cultured guinea pig scleral fibroblasts; however, their expression was confirmed in guinea pig liver. In addition, isolated cultured fibroblasts did not express integrin subunits αL, αM, and αX. This difference between results for cultured cells and intact sclera presumably reflects the presence in the latter of additional cell types. Both guinea pig scleral tissue and isolated scleral fibroblasts expressed all known integrin beta subunits. All results were verified through sequencing. The possible contributions of integrins to scleral remodeling make them plausible targets for myopia prevention. Data from this study will help guide future ex vivo and in vitro studies directed at understanding the relationship between scleral integrins and ocular growth regulation in the guinea pig model for myopia.

  4. Monoclonal antibodies to molluskan hemocyanin from Concholepas concholepas demonstrate common and specific epitopes among subunits.

    Science.gov (United States)

    Oliva, Harold; Moltedo, Bruno; De Ioannes, Pablo; Faunes, Fernando; De Ioannes, Alfredo E; Becker, María Inés

    2002-10-01

    We studied the reactivity of mouse monoclonal antibodies (MAbs) against the hemocyanin from the Chilean marine gastropod Concholepas concholepas (CCH). This protein has been successfully used as a carrier to produce antibodies to haptens and peptides. All MAbs (13) belonging to IgG subclass exhibit dissociation constants (K(d)) from 1 x 10(-7) M to 1 x 10(-9) M. MAbs were characterized by enzyme-linked immunosorbant assay (ELISA) using CCH treated with different procedures, including dissociation into CCH-A and CCH-B subunits, Western blot, enzymatic digestion, chemical deglycosylation, and thermal denaturation. MAbs were classified into three categories, according to subunit specificity by ELISA. The epitope distribution shows that CCH subunits display common epitopes (group I, 5 MAbs, 1H5, 2A8, 3A5, 3B3, and 3E3), as well as specific epitopes for CCH-A subunits (group II, 3 MAbs, 1B8, 4D8, and 8E5) and for CCH-B subunits (group III, 5 MAbs, 1A4, 1E4, 2H10, 3B7, and 7B4). The results can be summarized as follows: (1). six antibodies react with thermal denatured CCH, suggesting that they recognize linear epitopes, whereas seven recognize conformational epitopes; (2). oxidation of carbohydrate moieties does not affect the binding of the MAbs; (3). enzymatic digestion of CCH decreases the reactivity of all antibodies irrespective of the protease used (elastase or trypsin); (4). bringing together the above data, in addition to epitopic complementarity analysis, we identified 12 different epitopes on the CCH molecule recognized by these MAbs. The anti-CCH MAbs presented here can be useful tools to understand the subunit organization of the CCH and its complex structure, which can explain its immunogenic and immunostimulating properties in mammals.

  5. Three-dimensional crystals of ribosomes and their subunits from eu- and archaebacteria.

    Science.gov (United States)

    Glotz, C; Müssig, J; Gewitz, H S; Makowski, I; Arad, T; Yonath, A; Wittmann, H G

    1987-11-01

    Ordered three-dimensional crystals of 70S ribosomes as well as of 30S and 50S ribosomal subunits from various bacteria (E. coli, Bacillus stearothermophilus, Thermus thermophilus and Halobacterium marismortui) have been grown by vapour diffusion in hanging drops using mono- and polyalcohols. A new compact crystal form of 50S subunits has been obtained, and it is suitable for crystallographic studies at medium resolution. In addition, from one crystal form large crystals could be grown in X-ray capillaries. In all cases the crystals were obtained from functionally active ribosomal particles, and the particles from dissolved crystals retained their integrity and biological activity.

  6. Analysis of Maxi-K alpha subunit splice variants in human myometrium

    Directory of Open Access Journals (Sweden)

    Morrison John J

    2004-09-01

    Full Text Available Abstract Background Large-conductance, calcium-activated potassium (Maxi-K channels are implicated in the modulation of human uterine contractions and myometrial Ca2+ homeostasis. However, the regulatory mechanism(s governing the expression of Maxi-K channels with decreased calcium sensitivity at parturition are unclear. The objectives of this study were to investigate mRNA expression of the Maxi-K alpha subunit, and that of its splice variants, in human non-pregnant and pregnant myometrium, prior to and after labour onset, to determine whether altered expression of these splice variants is associated with decreased calcium sensitivity observed at labour onset. Methods Myometrial biopsies were obtained at hysterectomy (non-pregnant, NP, and at Caesarean section, at elective (pregnant not-in-labour, PNL and intrapartum (pregnant in-labour, PL procedures. RNA was extracted from all biopsies and quantitative real-time RT-PCR was used to investigate for possible differential expression of the Maxi-K alpha subunit, and that of its splice variants, between these functionally-distinct myometrial tissue sets. Results RT-PCR analysis identified the presence of a 132 bp and an 87 bp spliced exon of the Maxi-K alpha subunit in all three myometrial tissue sets. Quantitative real-time PCR indicated a decrease in the expression of the Maxi-K alpha subunit with labour onset. While there was no change in the proportion of Maxi-K alpha subunits expressing the 87 bp spliced exon, the proportion of alpha subunits expressing the 132 bp spliced exon was significantly increased with labour onset, compared to both non-pregnant and pregnant not-in-labour tissues. An increased proportion of 132 bp exon-containing alpha subunit variants with labour onset is of interest, as channels expressing this spliced exon have decreased calcium and voltage sensitivities. Conclusions Our findings suggest that decreased Maxi-K alpha subunit mRNA expression in human myometrium at

  7. Ribosome formation from subunits studied by stopped-flow and Rayleigh light scattering

    Directory of Open Access Journals (Sweden)

    Antoun Ayman

    2004-01-01

    Full Text Available Light scattering and standard stopped-flow techniques were used to monitor rapid association of ribosomal subunits during initiation of eubacterial protein synthesis. The effects of the initiation factors IF1, IF2, IF3 and buffer conditions on subunit association were studied along with the role of GTP in this process. The part of light scattering theory that is essential for kinetic measurements is high-lighted in the main text and a more general treatment of Rayleigh scattering from macromolecules is given in an appendix.

  8. The epithelial sodium channel γ-subunit is processed proteolytically in human kidney

    DEFF Research Database (Denmark)

    Langkilde, Rikke Zachar; Skjødt, Karsten; Marcussen, Niels

    2015-01-01

    The epithelial sodium channel (ENaC) of the kidney is necessary for extracellular volume homeostasis and normal arterial BP. Activity of ENaC is enhanced by proteolytic cleavage of the gamma-subunit and putative release of a 43-amino acid inhibitory tract from the gamma-subunit ectodomain. We......ENaC was detected consistently only in tissue from patients with proteinuria and observed in collecting ducts. In conclusion, human kidney gammaENaC is subject to proteolytic cleavage, yielding fragments compatible with furin cleavage, and proteinuria is associated with cleavage at the putative prostasin...

  9. Allosteric regulation and communication between subunits in uracil phosphoribosyltransferase from Sulfolobus solfataricus

    DEFF Research Database (Denmark)

    Arent, Susan; Harris, Pernille; Jensen, Kaj Frank

    2005-01-01

    organisms. To understand the allosteric regulation, crystal structures were determined for S. solfataricus UPRTase in complex with UMP and with UMP and the allosteric inhibitor CTP. Also, a structure with UMP bound in half of the active sites was determined. All three complexes form tetramers but reveal...... to rearrangements in the quaternary structure imply that this residue plays a major role in regulation of the enzyme and in communication between subunits. The ribose ring of UMP adopts alternative conformations in the cis and trans subunits of the UPRTase-UMP tetramer with associated differences...

  10. GABAA receptor subunit gene expression in human prefrontal cortex: comparison of schizophrenics and controls

    Science.gov (United States)

    Akbarian, S.; Huntsman, M. M.; Kim, J. J.; Tafazzoli, A.; Potkin, S. G.; Bunney, W. E. Jr; Jones, E. G.; Bloom, F. E. (Principal Investigator)

    1995-01-01

    The prefrontal cortex of schizophrenics is hypoactive and displays changes related to inhibitory, GABAergic neurons, and GABAergic synapses. These changes include decreased levels of glutamic acid decarboxylase (GAD), the enzyme for GABA synthesis, upregulation of muscimol binding, and downregulation of benzodiazepine binding to GABAA receptors. Studies in the visual cortex of nonhuman primates have demonstrated that gene expression for GAD and for several GABAA receptor subunit polypeptides is under control of neuronal activity, raising the possibility that similar mechanisms in the hypoactive prefrontal cortex of schizophrenics may explain the abnormalities in GAD and in GABAA receptor regulation. In the present study, which is the first of its type on human cerebral cortex, levels of mRNAs for six GABAA receptor subunits (alpha 1, alpha 2, alpha 5, beta 1, beta 2, gamma 2) and their laminar expression patterns were analyzed in the prefrontal cortex of schizophrenics and matched controls, using in situ hybridization histochemistry and densitometry. Three types of laminar expression pattern were observed: mRNAs for the alpha 1, beta 2, and gamma 2 subunits, which are the predominant receptor subunits expressed in the mature cortex, were expressed at comparatively high levels by cells of all six cortical layers, but most intensely by cells in lower layer III and layer IV. mRNAs for the alpha 2, alpha 5, and beta 1 subunits were expressed at lower levels; alpha 2 and beta 1 were expressed predominantly by cells in layers II, III, and IV; alpha 5 was expressed predominantly in layers IV, V, and VI. There were no significant changes in overall mRNA levels for any of the receptor subunits in the prefrontal cortex of schizophrenics, and the laminar expression pattern of all six receptor subunit mRNAs did not differ between schizophrenics and controls. Because gene expression for GABAA receptor subunits is not consistently altered in the prefrontal cortex of

  11. Subunit vaccine candidates against Aeromonas salmonicida in rainbow trout Oncorhynchus mykiss

    DEFF Research Database (Denmark)

    Marana, Moonika Haahr; Jørgensen, Louise von Gersdorff; Skov, Jakob

    2017-01-01

    rainbow trout (Oncorhynchus mykiss, Walbaum) aquaculture furunculosis outbreaks still occur. In this study we tested the efficacy of experimental subunit vaccines against A. salmonicida infection in rainbow trout. We utilized in silico screening of the proteome of A. salmonicida subsp. salmonicida strain...... A449 and identified potential protective protein antigens that were tested by in vivo challenge trial. A total of 14 proteins were recombinantly expressed in Escherichia coli and prepared in 3 different subunit vaccine combinations to immunize 3 groups of rainbow trout by intraperitoneal (i...

  12. Eukaryotic RNA polymerase subunit RPB8 is a new relative of the OB family.

    Science.gov (United States)

    Krapp, S; Kelly, G; Reischl, J; Weinzierl, R O; Matthews, S

    1998-02-01

    RNA polymerase II subunit RPB8 is an essential subunit that is highly conserved throughout eukaryotic evolution and is present in all three types of nuclear RNA polymerases. We report the first high resolution structural insight into eukaryotic RNA polymerase architecture with the solution structure of RPB8 from Saccharomyces cerevisiae. It consists of an eight stranded, antiparallel beta-barrel, four short helical regions and a large, unstructured omega-loop. The strands are connected in classic Greek-key fashion. The overall topology is unusual and contains a striking C2 rotational symmetry. Furthermore, it is most likely a novel associate of the oligonucleotide/oligosaccharide (OB) binding protein class.

  13. Activity-dependent control of NMDA receptor subunit composition at hippocampal mossy fibre synapses.

    Science.gov (United States)

    Carta, Mario; Srikumar, Bettadapura N; Gorlewicz, Adam; Rebola, Nelson; Mulle, Christophe

    2018-02-15

    CA3 pyramidal cells display input-specific differences in the subunit composition of synaptic NMDA receptors (NMDARs). Although at low density, GluN2B contributes significantly to NMDAR-mediated EPSCs at mossy fibre synapses. Long-term potentiation (LTP) of NMDARs triggers a modification in the subunit composition of synaptic NMDARs by insertion of GluN2B. GluN2B subunits are essential for the expression of LTP of NMDARs at mossy fibre synapses. Single neurons express NMDA receptors (NMDARs) with distinct subunit composition and biophysical properties that can be segregated in an input-specific manner. The dynamic control of the heterogeneous distribution of synaptic NMDARs is crucial to control input-dependent synaptic integration and plasticity. In hippocampal CA3 pyramidal cells from mice of both sexes, we found that mossy fibre (MF) synapses display a markedly lower proportion of GluN2B-containing NMDARs than associative/commissural synapses. The mechanism involved in such heterogeneous distribution of GluN2B subunits is not known. Here we show that long-term potentiation (LTP) of NMDARs, which is selectively expressed at MF-CA3 pyramidal cell synapses, triggers a modification in the subunit composition of synaptic NMDARs by insertion of GluN2B. This activity-dependent recruitment of GluN2B at mature MF-CA3 pyramidal cell synapses contrasts with the removal of GluN2B subunits at other glutamatergic synapses during development and in response to activity. Furthermore, although expressed at low levels, GluN2B is necessary for the expression of LTP of NMDARs at MF-CA3 pyramidal cell synapses. Altogether, we reveal a previously unknown activity-dependent regulation and function of GluN2B subunits that may contribute to the heterogeneous plasticity induction rules in CA3 pyramidal cells. © 2017 Centre Nationnal de la Recherche Scientifique. The Journal of Physiology © 2017 The Physiological Society.

  14. Family Environment and the Metabolic Syndrome: Results from the Hispanic Community Health Study/Study of Latinos (HCHS/SOL) Sociocultural Ancillary Study (SCAS).

    Science.gov (United States)

    Penedo, Frank J; Brintz, Carrie E; LLabre, Maria M; Arguelles, William; Isasi, Carmen R; Arredondo, Elva M; Navas-Nacher, Elena L; Perreira, Krista M; González, Hector M; Rodriguez, Carlos J; Daviglus, Martha; Schneiderman, Neil; Gallo, Linda C

    2015-12-01

    Metabolic syndrome (MetS) is a risk factor for cardiovascular disease. Very limited work has evaluated associations of sociocultural processes with prevalence of the MetS. The purpose of the present study was to evaluate associations between family environment (cohesion/conflict) and the MetS, in a multi-site sample of US Hispanics/Latinos. A total of 3278 participants from the Hispanic Community Health Study/Study of Latinos underwent a clinical exam and completed psychosocial measures including family environment (cohesion and conflict) as part of the Sociocultural Ancillary Study. The association between family environment and the MetS was moderated by sex. Among all women, higher family conflict was associated with MetS prevalence. Results by ancestry group showed that only among Cuban women, higher conflict was associated with the MetS, whereas only among Dominican men, greater cohesion was associated with the MetS. The family context may be a sociocultural protective or risk factor among Hispanics/Latinos in terms of MetS risk, but these associations may vary by sex and Hispanic background.

  15. The potential of liquid-based cytology in lymph node cytological evaluation: the role of morphology and the aid of ancillary techniques.

    Science.gov (United States)

    Rossi, E D; Martini, M; Straccia, P; Bizzarro, T; Fadda, G; Larocca, L M

    2016-02-01

    Our aim was to evaluate the feasibility and diagnostic accuracy of liquid-based cytology (LBC) on lymph node fine needle aspiration (FNA). FNA may fulfil a challenging role in the evaluation of the majority of primary (benign and malignant) diagnoses as well as metastatic lymph node lesions. Although the morphological features may be quite easily recognized, cytological samples with a scant cellular component may raise some issues. We appraised 263 cytological lymph nodes from different body regions analysed between January and December 2013, including 137 male and 126 female patients, and processed with LBC. The cytological diagnoses included 160 benign and 103 malignant lesions. We reported 35 benign and 73 malignant lesions from 108 with surgical follow-up. The latter malignant series included 68 metastatic lesions, four suspicious for malignancy and one inadequate sample. The cytological diagnoses were supported by 62 conclusive immunocytochemical and 28 molecular analyses. Of the 108 cases, we documented 35 true negatives, 72 true positives, one false negative and no false positives, resulting in 98.6% sensitivity, 100% specificity, 99% diagnostic accuracy, 97.2% negative predictive value and 100% positive predictive value. FNA represents the first diagnostic tool in lymph node management and a reliable approach in order to avoid an excision biopsy. Furthermore, LBC is a feasible method for ancillary tests for which methanol-fixed samples are suitable, such as immunocytochemistry and molecular analysis. © 2014 John Wiley & Sons Ltd.

  16. Validation of interpersonal support evaluation list-12 (ISEL-12) scores among English- and Spanish-speaking Hispanics/Latinos from the HCHS/SOL Sociocultural Ancillary Study.

    Science.gov (United States)

    Merz, Erin L; Roesch, Scott C; Malcarne, Vanessa L; Penedo, Frank J; Llabre, Maria M; Weitzman, Orit B; Navas-Nacher, Elena L; Perreira, Krista M; Gonzalez, Franklyn; Ponguta, Liliana A; Johnson, Timothy P; Gallo, Linda C

    2014-06-01

    The Interpersonal Support Evaluation List-12 (ISEL-12; Cohen, Mermelstein, Kamarck, & Hoberman, 1985) is broadly employed as a short-form measure of the traditional ISEL, which measures functional (i.e., perceived) social support. The ISEL-12 can be scored by summing the items to create an overall social support score; three subscale scores representing appraisal, belonging, and tangible social support have also been proposed. Despite extensive use, studies of the psychometric properties of ISEL-12 scores have been limited, particularly among Hispanics/Latinos, the largest and fastest growing ethnic group in the United States. The current study investigated the reliability and structural and convergent validity of ISEL-12 scores using data from 5,313 Hispanics/Latinos who participated in the Hispanic Community Health Study/Study of Latinos Sociocultural Ancillary Study. Participants completed measures in English or Spanish and identified their ancestry as Dominican, Central American, Cuban, Mexican, Puerto Rican, or South American. Cronbach's alphas suggested adequate internal consistency for the total score for all languages and ancestry groups; coefficients for the subscale scores were not acceptable. Confirmatory factor analyses revealed that the one-factor and three-factor models fit the data equally well. Results from multigroup confirmatory factor analyses supported a similar one-factor structure with equivalent response patterns and variances between language groups and ancestry groups. Convergent validity analyses suggested that the total social support score related to scores of social network integration, life engagement, perceived stress, and negative affect (depression, anxiety) in the expected directions.

  17. Machismo, Marianismo, and Negative Cognitive-Emotional Factors: Findings From the Hispanic Community Health Study/Study of Latinos Sociocultural Ancillary Study.

    Science.gov (United States)

    Nuñez, Alicia; González, Patricia; Talavera, Gregory A; Sanchez-Johnsen, Lisa; Roesch, Scott C; Davis, Sonia M; Arguelles, William; Womack, Veronica Y; Ostrovsky, Natania W; Ojeda, Lizette; Penedo, Frank J; Gallo, Linda C

    2016-11-01

    There is limited research on the traditional Hispanic male and female gender roles of machismo and marianismo, respectively, in relation to negative cognitions and emotions. Given the vulnerability of Hispanics to negative cognitions and emotions, it is important to examine sociocultural correlates of emotional distress. Therefore, we examined associations of machismo and marianismo with negative cognitive-emotional factors (i.e., depression symptoms; cynical hostility; and trait anxiety and anger) in the Hispanic Community Health Study/Study of Latinos Sociocultural Ancillary Study, a cross-sectional cohort study of sociocultural and psychosocial correlates of cardiometabolic health. Participants were aged 18-74 years and self-identified as Hispanic of Central American, Cuban, Dominican, Mexican, Puerto Rican, South American, and other Hispanic background ( N = 4,426). Results revealed that specific components of machismo (traditional machismo) and marianismo (family and spiritual pillar dimensions) were associated with higher levels of negative cognitions and emotions after adjusting for socio-demographic factors ( p < .05); these associations remained consistent across sex, Hispanic background group, and acculturation. Findings can inform mental health interventions and contribute to our understanding of the importance of gender role socialization in the context of self-reported negative cognitive-emotional factors in Hispanics.

  18. Structural basis of subunit selectivity for competitive NMDA receptor antagonists with preference for GluN2A over GluN2B subunits

    Energy Technology Data Exchange (ETDEWEB)

    Lind, Genevieve E.; Mou, Tung-Chung; Tamborini, Lucia; Pomper, Martin G.; De Micheli, Carlo; Conti, Paola; Pinto, Andrea; Hansen, Kasper B. (JHU); (Milan); (Montana)

    2017-07-31

    NMDA-type glutamate receptors are ligand-gated ion channels that contribute to excitatory neurotransmission in the central nervous system (CNS). Most NMDA receptors comprise two glycine-binding GluN1 and two glutamate-binding GluN2 subunits (GluN2A–D). We describe highly potent (S)-5-[(R)-2-amino-2-carboxyethyl]-4,5-dihydro-1H-pyrazole-3-carboxylic acid (ACEPC) competitive GluN2 antagonists, of which ST3 has a binding affinity of 52 nM at GluN1/2A and 782 nM at GluN1/2B receptors. This 15-fold preference of ST3 for GluN1/2A over GluN1/2B is improved compared with NVP-AAM077, a widely used GluN2A-selective antagonist, which we show has 11-fold preference for GluN1/2A over GluN1/2B. Crystal structures of the GluN1/2A agonist binding domain (ABD) heterodimer with bound ACEPC antagonists reveal a binding mode in which the ligands occupy a cavity that extends toward the subunit interface between GluN1 and GluN2A ABDs. Mutational analyses show that the GluN2A preference of ST3 is primarily mediated by four nonconserved residues that are not directly contacting the ligand, but positioned within 12 Å of the glutamate binding site. Two of these residues influence the cavity occupied by ST3 in a manner that results in favorable binding to GluN2A, but occludes binding to GluN2B. Thus, we reveal opportunities for the design of subunit-selective competitive NMDA receptor antagonists by identifying a cavity for ligand binding in which variations exist between GluN2A and GluN2B subunits. This structural insight suggests that subunit selectivity of glutamate-site antagonists can be mediated by mechanisms in addition to direct contributions of contact residues to binding affinity.

  19. The crystal structure of the complex of Zea mays alpha subunit with a fragment of human beta subunit provides the clue to the architecture of protein kinase CK2 holoenzyme

    DEFF Research Database (Denmark)

    Battistutta, R; Sarno, S; De Moliner, E

    2000-01-01

    The crystal structure of a complex between the catalytic alpha subunit of Zea mays CK2 and a 23-mer peptide corresponding the C-terminal sequence 181-203 of the human CK2 regulatory beta subunit has been determined at 3.16-A resolution. The complex, composed of two alpha chains and two peptides, ...

  20. Cholinergic cells in the nucleus basalis of mice express the N-methyl-D-aspartate-receptor subunit NR2C and its replacement by the NR2B subunit enhances frontal and amygdaloid acetylcholine levels

    NARCIS (Netherlands)

    De Souza Silva, M. A.; Dolga, Amalia; Pieri, I.; Marchetti, L.; Eisel, U. L. M.; Huston, J. P.; Dere, E.

    2006-01-01

    It is known that glutamatergic and cholinergic systems interact functionally at the level of the cholinergic basal forebrain. The N-methyl-D-aspartate receptor (NMDA-R) is a multiprotein complex composed of NR1, NR2 and/or NR3 subunits. The subunit composition of NMDA-R of cholinergic cells in the

  1. Purification and functional reconstitution of a seven-subunit mrp-type na+/h+ antiporter.

    Science.gov (United States)

    Morino, Masato; Suzuki, Toshiharu; Ito, Masahiro; Krulwich, Terry Ann

    2014-01-01

    Mrp antiporters and their homologues in the cation/proton antiporter 3 family of the Membrane Transporter Database are widely distributed in bacteria. They have major roles in supporting cation and cytoplasmic pH homeostasis in many environmental, extremophilic, and pathogenic bacteria. These antiporters require six or seven hydrophobic proteins that form hetero-oligomeric complexes, while most other cation/proton antiporters require only one membrane protein for their activity. The resemblance of three Mrp subunits to membrane-embedded subunits of the NADH:quinone oxidoreductase of respiratory chains and to subunits of several hydrogenases has raised interest in the evolutionary path and commonalities of their proton-translocating domains. In order to move toward a greater mechanistic understanding of these unusual antiporters and to rigorously demonstrate that they function as secondary antiporters, powered by an imposed proton motive force, we established a method for purification and functional reconstitution of the seven-subunit Mrp antiporter from alkaliphilic Bacillus pseudofirmus OF4. Na(+)/H(+) antiporter activity was demonstrated by a fluorescence-based assay with proteoliposomes in which the Mrp complex was coreconstituted with a bacterial FoF1-ATPase. Proton pumping by the ATPase upon addition of ATP generated a proton motive force across the membranes that powered antiporter activity upon subsequent addition of Na(+).

  2. Block of nicotinic acetylcholine receptors by philanthotoxins is strongly dependent on their subunit composition

    DEFF Research Database (Denmark)

    Kachel, Hamid S; Patel, Rohit N; Franzyk, Henrik

    2016-01-01

    -fold selectivity of PhTX-12 over PhTX-343 for embryonic muscle-type nicotinic acetylcholine receptors (nAChRs) in TE671 cells. We investigated their inhibition of different neuronal nAChR subunit combinations as well as of embryonic muscle receptors expressed in Xenopus oocytes. Whole-cell currents...

  3. Differential regulation by AMP and ADP of AMPK complexes containing different γ subunit isoforms

    DEFF Research Database (Denmark)

    Ross, Fiona A; Jensen, Thomas Elbenhardt; Hardie, D Grahame

    2016-01-01

    The g subunits of heterotrimeric AMPK complexes contain the binding sites for the regulatory adenine nucleotides AMP, ADP and ATP. We addressed whether complexes containing different g isoforms display different responses to adenine nucleotides by generating cells stably expressing FLAG-tagged ve...

  4. Recombinant cholera toxin B subunit in Escherichia coli: high-level secretion, purification, and characterization

    NARCIS (Netherlands)

    Slos, P.; Speck, D.; Accart, N.; Kolbe, H.V.; Schubnel, D.; Bouchon, B.; Bischoff, Rainer; Kieny, M.P.

    1994-01-01

    The gene coding for cholera toxin subunit B (CT-B) was fused to a modified ompA signal sequence and subsequently cloned into a high expression vector based on the regulatory signals of the arabinose operon of Salmonella typhimurium. Upon induction of gene expression in Escherichia coli, a product of

  5. epsilon, a New Subunit of RNA Polymerase Found in Gram-Positive Bacteria

    Czech Academy of Sciences Publication Activity Database

    Keller, A. N.; Yang, X.; Wiedermannová, Jana; Delumeau, O.; Krásný, Libor; Lewis, P. J.

    2014-01-01

    Roč. 196, č. 20 (2014), s. 3622-3632 ISSN 0021-9193 R&D Projects: GA ČR(CZ) GBP305/12/G034 Institutional support: RVO:61388971 Keywords : RNA polymerase * subunit * X-ray crystallography Subject RIV: EE - Microbiology, Virology Impact factor: 2.808, year: 2014

  6. Stability of influenza sub-unit vaccine. Does a couple of days outside the refrigerator matter?

    NARCIS (Netherlands)

    Coenen, F; Tolboom, J T B M; Frijlink, H W

    2006-01-01

    In this study 27 full scale production batches of influenza sub-unit vaccine were evaluated on their stability. The batches varied with respect to the strains they contained and regarding the presence of the preservative thiomersal in the solution. The stability study showed that haemagglutinin

  7. A CK2 site is reversibly phosphorylated in the photosystem II subunit CP29

    NARCIS (Netherlands)

    Testi, Maria Grazia; Croce, Roberta; Polverino-De Laureto, Patrizia; Bassi, Roberto

    1996-01-01

    Protein phosphorylation is a major mechanism in the regulation of protein function. In chloroplast thylakoids several photosystem II subunits, including the major antenna light-harvesting complex II and several core complex components, are reversibly phosphorylated depending on the redox state of

  8. The δ subunit of RNA polymerase guides promoter selectivity and virulence in Staphylococcus aureus.

    Science.gov (United States)

    Weiss, Andy; Ibarra, J Antonio; Paoletti, Jessica; Carroll, Ronan K; Shaw, Lindsey N

    2014-04-01

    In Gram-positive bacteria, and particularly the Firmicutes, the DNA-dependent RNA polymerase (RNAP) complex contains an additional subunit, termed the δ factor, or RpoE. This enigmatic protein has been studied for more than 30 years for various organisms, but its function is still not well understood. In this study, we investigated its role in the major human pathogen Staphylococcus aureus. We showed conservation of important structural regions of RpoE in S. aureus and other species and demonstrated binding to core RNAP that is mediated by the β and/or β' subunits. To identify the impact of the δ subunit on transcription, we performed transcriptome sequencing (RNA-seq) analysis and observed 191 differentially expressed genes in the rpoE mutant. Ontological analysis revealed, quite strikingly, that many of the downregulated genes were known virulence factors, while several mobile genetic elements (SaPI5 and prophage SA3usa) were strongly upregulated. Phenotypically, the rpoE mutant had decreased accumulation and/or activity of a number of key virulence factors, including alpha toxin, secreted proteases, and Panton-Valentine leukocidin (PVL). We further observed significantly decreased survival of the mutant in whole human blood, increased phagocytosis by human leukocytes, and impaired virulence in a murine model of infection. Collectively, our results demonstrate that the δ subunit of RNAP is a critical component of the S. aureus transcription machinery and plays an important role during infection.

  9. Development of haplotype-specific molecular markers for the low-molecular-weight glutenin subunits

    Science.gov (United States)

    Low-molecular-weight glutenin subunits (LMW-GSs) are one of the major components of gluten and their allelic variation has been widely associated with numerous wheat end-use quality parameters. These proteins are encoded by multigene families located at the orthologous Glu-3 loci (Glu-A3, Glu-B3 and...

  10. An immune stimulating complex (iscom) subunit rabies vaccine protects dogs and mice against street rabies challenge.

    NARCIS (Netherlands)

    M. Fekadu; J.H. Schaddock; J. Ekströ m; A.D.M.E. Osterhaus (Albert); D.W. Sanderlin; B. Sundquist; B. Morein (Bror)

    1992-01-01

    textabstractDogs and mice were immunized with either a rabies glycoprotein subunit vaccine incorporated into an immune stimulating complex (ISCOM) or a commercial human diploid cell vaccine (HDCV) prepared from a Pitman Moore (PM) rabies vaccine strain. Pre-exposure vaccination of mice with two

  11. Reconstitution of normal and hyperactivated forms of casein kinase-2 by variably mutated beta-subunits

    DEFF Research Database (Denmark)

    Boldyreff, B; Meggio, F; Pinna, L A

    1993-01-01

    Twenty-one mutants of the noncatalytic beta-subunit of human casein kinase-2 have been created, expressed in Escherichia coli, and purified to homogeneity. They are either modified at the autophosphorylation site (mutants beta delta 1-4 and beta A 5,6) or bear variable deletions in their C...

  12. Binary Toxin Subunits of Lysinibacillus sphaericus Are Monomeric and Form Heterodimers after In Vitro Activation.

    Directory of Open Access Journals (Sweden)

    Wahyu Surya

    Full Text Available The binary toxin from Lysinibacillus sphaericus has been successfully used for controlling mosquito-transmitted diseases. An activation step shortens both subunits BinA and BinB before their interaction with membranes and internalization in midgut cells, but the precise role of this activation step is unknown. Herein, we show conclusively using three orthogonal biophysical techniques that protoxin subunits form only monomers in aqueous solution. However, in vitro activated toxins readily form heterodimers. This oligomeric state did not change after incubation of these heterodimers with detergent. These results are consistent with the evidence that maximal toxicity in mosquito larvae is achieved when the two subunits, BinA and BinB, are in a 1:1 molar ratio, and directly link proteolytic activation to heterodimerization. Formation of a heterodimer must thus be necessary for subsequent steps, e.g., interaction with membranes, or with a suitable receptor in susceptible mosquito species. Lastly, despite existing similarities between BinB C-terminal domain with domains 3 and 4 of pore-forming aerolysin, no aerolysin-like SDS-resistant heptameric oligomers were observed when the activated Bin subunits were incubated in the presence of detergents or lipidic membranes.

  13. Structural properties of a peptide derived from H+-V-ATPase subunit a

    NARCIS (Netherlands)

    Vermeer, L.S.; Reat, V.; Hemminga, M.A.; Milon, A.

    2009-01-01

    The 3D structure of a peptide derived from the putative transmembrane segment 7 (TM7) of subunit a from H+-V-ATPase from Saccharomyces cerevisiae has been determined by solution state NMR in SDS. A stable helix is formed from L736 up to and including Q745, the lumenal half of the putative TM7. The

  14. Positive modulation of delta-subunit containing GABAA receptors in mouse neurons

    DEFF Research Database (Denmark)

    Vardya, Irina; Hoestgaard-Jensen, Kirsten; Nieto-Gonzalez, Jose Luis

    2012-01-01

    δ-subunit containing extrasynaptic GABA(A) receptors are potential targets for modifying neuronal activity in a range of brain disorders. With the aim of gaining more insight in synaptic and extrasynaptic inhibition, we used a new positive modulator, AA29504, of δ-subunit containing GABA(A) recep......δ-subunit containing extrasynaptic GABA(A) receptors are potential targets for modifying neuronal activity in a range of brain disorders. With the aim of gaining more insight in synaptic and extrasynaptic inhibition, we used a new positive modulator, AA29504, of δ-subunit containing GABA......(A) receptors in mouse neurons in vitro and in vivo. Whole-cell patch-clamp recordings were carried out in the dentate gyrus in mouse brain slices. In granule cells, AA29504 (1 μM) caused a 4.2-fold potentiation of a tonic current induced by THIP (1 μM), while interneurons showed a potentiation of 2.6-fold......-free environment using Ca²⁺ imaging in cultured neurons, AA29504 showed GABA(A) receptor agonism in the absence of agonist. Finally, AA29504 exerted dose-dependent stress-reducing and anxiolytic effects in mice in vivo. We propose that AA29504 potentiates δ-containing GABA(A) receptors to enhance tonic inhibition...

  15. A comparative study of ATPase subunit 9 (Atp9) gene between ...

    African Journals Online (AJOL)

    ATPase subunit 9 gene (Atp9) is an important functional gene in mitochondria, and is closely related with energy supply. RNA editing of atp9 gene was associated with male sterility in plants. In this study, the atp9 gene in soybeans was cloned from a soybean cytoplasmic male sterile line NJCMS2A and its maintainer line ...

  16. The thermal structural transition of alpha-crystallin modulates subunit interactions and increases protein solubility.

    Directory of Open Access Journals (Sweden)

    Giuseppe Maulucci

    Full Text Available BACKGROUND: Alpha crystallin is an oligomer composed of two types of subunits, alpha-A and alpha-B crystallin, and is the major constituent of human lens. The temperature induced condensation of alpha-crystallin, the main cause for eye lens opacification (cataract, is a two step-process, a nucleation followed by an aggregation phase, and a protective effect towards the aggregation is exhibited over the alpha crystallin phase transition temperature (Tc = 318.16 K. METHODS/RESULTS: To investigate if a modulation of the subunit interactions over Tc could trigger the protective mechanism towards the aggregation, we followed, by using simultaneously static and dynamic light scattering, the temperature induced condensation of alpha-crystallin. By developing a mathematical model able to uncouple the nucleation and aggregation processes, we find a previously unobserved transition in the nucleation rate constant. Its temperature dependence allows to determine fundamental structural parameters, the chemical potential (Δμ and the interfacial tension (γ of the aggregating phase, that characterize subunit interactions. CONCLUSIONS/GENERAL SIGNIFICANCE: The decrease of both Δμ and γ at Tc, and a relative increase in solubility, reveal a significative decrease in the strenght of alpha-crystallin subunits interactions, which protects from supramolecolar condensation in hypertermic conditions. On the whole, we suggest a general approach able to understand the structural and kinetic mechanisms involved in aggregation-related diseases and in drugs development and testing.

  17. The Cac2 subunit is essential for productive histone binding and nucleosome assembly in CAF-1

    Energy Technology Data Exchange (ETDEWEB)

    Mattiroli, Francesca; Gu, Yajie; Balsbaugh, Jeremy L.; Ahn, Natalie G.; Luger, Karolin

    2017-04-18

    Nucleosome assembly following DNA replication controls epigenome maintenance and genome integrity. Chromatin assembly factor 1 (CAF-1) is the histone chaperone responsible for histone (H3-H4)2 deposition following DNA synthesis. Structural and functional details for this chaperone complex and its interaction with histones are slowly emerging. Using hydrogen-deuterium exchange coupled to mass spectrometry, combined with in vitro and in vivo mutagenesis studies, we identified the regions involved in the direct interaction between the yeast CAF-1 subunits, and mapped the CAF-1 domains responsible for H3-H4 binding. The large subunit, Cac1 organizes the assembly of CAF-1. Strikingly, H3-H4 binding is mediated by a composite interface, shaped by Cac1-bound Cac2 and the Cac1 acidic region. Cac2 is indispensable for productive histone binding, while deletion of Cac3 has only moderate effects on H3-H4 binding and nucleosome assembly. These results define direct structural roles for yeast CAF-1 subunits and uncover a previously unknown critical function of the middle subunit in CAF-1.

  18. Unassigned MURF1 of kinetoplastids codes for NADH dehydrogenase subunit 2

    Directory of Open Access Journals (Sweden)

    Burger Gertraud

    2008-10-01

    Full Text Available Abstract Background In a previous study, we conducted a large-scale similarity-free function prediction of mitochondrion-encoded hypothetical proteins, by which the hypothetical gene murf1 (maxicircle unidentified reading frame 1 was assigned as nad2, encoding subunit 2 of NADH dehydrogenase (Complex I of the respiratory chain. This hypothetical gene occurs in the mitochondrial genome of kinetoplastids, a group of unicellular eukaryotes including the causative agents of African sleeping sickness and leishmaniasis. In the present study, we test this assignment by using bioinformatics methods that are highly sensitive in identifying remote homologs and confront the prediction with available biological knowledge. Results Comparison of MURF1 profile Hidden Markov Model (HMM against function-known profile HMMs in Pfam, Panther and TIGR shows that MURF1 is a Complex I protein, but without specifying the exact subunit. Therefore, we constructed profile HMMs for each individual subunit, using all available sequences clustered at various identity thresholds. HMM-HMM comparison of these individual NADH subunits against MURF1 clearly identifies this hypothetical protein as NAD2. Further, we collected the relevant experimental information about kinetoplastids, which provides additional evidence in support of this prediction. Conclusion Our in silico analyses provide convincing evidence for MURF1 being a highly divergent member of NAD2.

  19. Expression, purification and crystallization of the catalytic subunit of protein kinase CK2 from Zea mays

    DEFF Research Database (Denmark)

    Guerra, B; Niefind, K; Pinna, L A

    1998-01-01

    The catalytic (alpha) subunit of protein kinase CK2 (CK2alpha) was originally cloned and overexpressed in the Escherichia coli strain pT7-7/BL21(DE3). The protein has been purified to homogeneity and crystallized. The crystals belong to the monoclinic space group C2, they have unit-cell parameter...

  20. Domain interactions of the peripheral preprotein translocase subunit SecA

    NARCIS (Netherlands)

    Blaauwen, T.den; Fekkes, P.; de Wit, J.G.; Kuiper, W.; Driessen, A.J.M.

    1996-01-01

    The homodimeric SecA protein is the peripheral subunit of the preprotein translocase in bacteria. It binds the preprotein and promotes its translocation across the bacterial cytoplasmic membrane by nucleotide modulated coinsertion and deinsertion into the membrane, SecA has two essential nucleotide

  1. Domain dynamics of the Bacillus subtilis peripheral preprotein translocase subunit SecA

    NARCIS (Netherlands)

    Driessen, A.J.M.; Ladbury, JE; Chowdhry, BZ

    1998-01-01

    The homodimeric SecA protein is the peripheral subunit of the preprotein translocase in bacteria. It promotes the preprotein translocation across the cytoplasmic membrane by nucleotide-modulated co-insertion and de-insertion into the integral domain of the translocase. SecA has two essential

  2. Asymmetric expression of protein kinase CK2 subunits in human kidney tumors

    DEFF Research Database (Denmark)

    Stalter, G; Siemer, S; Becht, E

    1994-01-01

    of protein kinase CK2 alpha in tumors/normal tissue (T/N) was 1.58 and that of the protein kinase CK2 beta (T/N) was 2.65. The data suggest that the generally described increase in protein kinase CK2 activity in tumor cells may to some extent result from a deregulation in subunit biosynthesis or degradation...

  3. The TFIIH Subunit p89 (XPB Localizes to the Centrosome during Mitosis

    Directory of Open Access Journals (Sweden)

    Achim Weber

    2010-01-01

    Full Text Available Background: The general transcription factor II H (TFIIH, comprised of a core complex and an associated CAK-complex, functions in transcription, DNA repair and cell cycle control. Mutations of the two largest subunits, p89 (XPB and p80 (XPD, cause the hereditary cancer-prone syndrome xeroderma pigmentosum.

  4. Production of a highly immunogenic subunit ISCOM vaccine against Bovine Viral Diarrhea Virus

    DEFF Research Database (Denmark)

    Kamstrup, Søren; Roensholt, L.; Jensen, M.Holm

    1999-01-01

    by Vaccination of the dam. We describe in this report the production and initial testing of an inactivated subunit vaccine against BVDV. The vaccine is based on production of antigen in primary bovine cell cultures, extraction of antigens from infected cells with detergent, chromatographic purification...

  5. Over-production, renaturation and reconstitution of delta and epsilon subunits from chloroplast and cyanobacterial F1

    NARCIS (Netherlands)

    Steinemann, D.; Lill, H; Junge, Wolfgang; Engelbrecht, Siegfried

    1994-01-01

    We studied the functioning of chimeric F0F1-ATPases by replacing subunits delta and epsilon of spinach CF1 with their counterparts from Synechocystis sp. PCC 6803. The sequence identities between these subunits are 26 and 41%, respectively. For a systematic approach to such studies and later

  6. Role of the Rubisco small subunit. Final report for period May 1, 1997--April 30,2000

    Energy Technology Data Exchange (ETDEWEB)

    Spreitzer, Robert J.

    2000-10-04

    CO{sub 2} and O{sub 2} are mutually competitive at the active site of ribulose-1,5-biphosphate (RuBP) carboxylase/oxygenase (Rubisco). Rubisco contains two subunits, each present in eight copies. The 15-kD small subunit is coded by a family of nuclear RbcS genes. Until now, the role of the small subunit in Rubisco structure or catalytic efficiency is not known. Because of other work in eliminating the two RbcS genes in the green algo Chlamydomonas reinhardtii, it is now possible to address questions about the structure-function relationships of the eukaryotic small subunit. There are three specific aims in this project: (1) Alanine scanning mutagenesis is being used to dissect the importance of the {beta}A/{beta}B loop, a feature unique to the eukaryotic small subunit. (2) Random mutagenesis is being used to identify additional residues or regions of the small subunit that are important for holoenzyme assembly and function. (3) Attempts are being made to express foreign small subunits in Chlamydomonas to examine the contribution of small subunits to holoenzyme assembly, catalytic efficiency, and CO{sub 2}/O{sub 2} specificity.

  7. Extricating Manual and Non-Manual Features for Subunit Level Medical Sign Modelling in Automatic Sign Language Classification and Recognition.

    Science.gov (United States)

    R, Elakkiya; K, Selvamani

    2017-09-22

    Subunit segmenting and modelling in medical sign language is one of the important studies in linguistic-oriented and vision-based Sign Language Recognition (SLR). Many efforts were made in the precedent to focus the functional subunits from the view of linguistic syllables but the problem is implementing such subunit extraction using syllables is not feasible in real-world computer vision techniques. And also, the present recognition systems are designed in such a way that it can detect the signer dependent actions under restricted and laboratory conditions. This research paper aims at solving these two important issues (1) Subunit extraction and (2) Signer independent action on visual sign language recognition. Subunit extraction involved in the sequential and parallel breakdown of sign gestures without any prior knowledge on syllables and number of subunits. A novel Bayesian Parallel Hidden Markov Model (BPaHMM) is introduced for subunit extraction to combine the features of manual and non-manual parameters to yield better results in classification and recognition of signs. Signer independent action aims in using a single web camera for different signer behaviour patterns and for cross-signer validation. Experimental results have proved that the proposed signer independent subunit level modelling for sign language classification and recognition has shown improvement and variations when compared with other existing works.

  8. Basal Levels of AMPA Receptor GluA1 Subunit Phosphorylation at Threonine 840 and Serine 845 in Hippocampal Neurons

    Science.gov (United States)

    Babiec, Walter E.; Guglietta, Ryan; O'Dell, Thomas J.

    2016-01-01

    Dephosphorylation of AMPA receptor (AMPAR) GluA1 subunits at two sites, serine 845 (S845) and threonine 840 (T840), is thought to be involved in NMDA receptor-dependent forms of long-term depression (LTD). Importantly, the notion that dephosphorylation of these sites contributes to LTD assumes that a significant fraction of GluA1 subunits are…

  9. Subunit–subunit interactions are weakened in mutant forms of acetohydroxy acid synthase insensitive to valine inhibition

    Czech Academy of Sciences Publication Activity Database

    Kyselková, Martina; Janata, Jiří; Ságová-Marečková, M.; Kopecký, J.

    2010-01-01

    Roč. 192, č. 3 (2010), s. 195-200 ISSN 0302-8933 R&D Projects: GA MŠk 2B08064 Institutional research plan: CEZ:AV0Z50200510 Keywords : Streptomyces cinnamonensis * Acetohydroxy acid synthase * Subunit-subunit interaction Subject RIV: EE - Microbiology, Virology Impact factor: 1.754, year: 2010

  10. Distribution of the a2, a3, and a5 nicotinic acetylcholine receptor subunits in the chick brain

    Directory of Open Access Journals (Sweden)

    Torrão A.S.

    1997-01-01

    Full Text Available Nicotinic acetylcholine receptors (nAChRs are ionotropic receptors comprised of a and ß subunits. These receptors are widely distributed in the central nervous system, and previous studies have revealed specific patterns of localization for some nAChR subunits in the vertebrate brain. In the present study we used immunohistochemical methods and monoclonal antibodies to localize the a2, a3, and a5 nAChR subunits in the chick mesencephalon and diencephalon. We observed a differential distribution of these three subunits in the chick brain, and showed that the somata and neuropil of many central structures contain the a5 nAChR subunit. The a2 and a3 subunits, on the other hand, exhibited a more restricted distribution than a5 and other subunits previously studied, namely a7, a8 and ß2. The patterns of distribution of the different nAChR subunits suggest that neurons in many brain structures may contain several subtypes of nAChRs and that in a few regions one particular subtype may determine the cholinergic nicotinic responses

  11. Construction of a hybrid β-hexosaminidase subunit capable of forming stable homodimers that hydrolyze GM2 ganglioside in vivo

    Directory of Open Access Journals (Sweden)

    Michael B Tropak

    2016-01-01

    Full Text Available Tay-Sachs or Sandhoff disease result from mutations in either the evolutionarily related HEXA or HEXB genes encoding respectively, the α- or β-subunits of β-hexosaminidase A (HexA. Of the three Hex isozymes, only HexA can interact with its cofactor, the GM2 activator protein (GM2AP, and hydrolyze GM2 ganglioside. A major impediment to establishing gene or enzyme replacement therapy based on HexA is the need to synthesize both subunits. Thus, we combined the critical features of both α- and β-subunits into a single hybrid µ-subunit that contains the α-subunit active site, the stable β-subunit interface and unique areas in each subunit needed to interact with GM2AP. To facilitate intracellular analysis and the purification of the µ-homodimer (HexM, CRISPR-based genome editing was used to disrupt the HEXA and HEXB genes in a Human Embryonic Kidney 293 cell line stably expressing the µ-subunit. In association with GM2AP, HexM was shown to hydrolyze a fluorescent GM2 ganglioside derivative both in cellulo and in vitro. Gene transfer studies in both Tay-Sachs and Sandhoff mouse models demonstrated that HexM expression reduced brain GM2 ganglioside levels.

  12. Construction of a hybrid β-hexosaminidase subunit capable of forming stable homodimers that hydrolyze GM2 ganglioside in vivo

    Science.gov (United States)

    Tropak, Michael B; Yonekawa, Sayuri; Karumuthil-Melethil, Subha; Thompson, Patrick; Wakarchuk, Warren; Gray, Steven J; Walia, Jagdeep S; Mark, Brian L; Mahuran, Don

    2016-01-01

    Tay-Sachs or Sandhoff disease result from mutations in either the evolutionarily related HEXA or HEXB genes encoding respectively, the α- or β-subunits of β-hexosaminidase A (HexA). Of the three Hex isozymes, only HexA can interact with its cofactor, the GM2 activator protein (GM2AP), and hydrolyze GM2 ganglioside. A major impediment to establishing gene or enzyme replacement therapy based on HexA is the need to synthesize both subunits. Thus, we combined the critical features of both α- and β-subunits into a single hybrid µ-subunit that contains the α-subunit active site, the stable β-subunit interface and unique areas in each subunit needed to interact with GM2AP. To facilitate intracellular analysis and the purification of the µ-homodimer (HexM), CRISPR-based genome editing was used to disrupt the HEXA and HEXB genes in a Human Embryonic Kidney 293 cell line stably expressing the µ-subunit. In association with GM2AP, HexM was shown to hydrolyze a fluorescent GM2 ganglioside derivative both in cellulo and in vitro. Gene transfer studies in both Tay-Sachs and Sandhoff mouse models demonstrated that HexM expression reduced brain GM2 ganglioside levels. PMID:26966698

  13. A formalism for scattering of complex composite structures. I. Applications to branched structures of asymmetric sub-units

    DEFF Research Database (Denmark)

    Svaneborg, Carsten; Pedersen, Jan Skov

    2012-01-01

    to structural connectivity is completely decoupled from internal structure of the sub-units. This allows sub-units to be replaced by more complex structures. We illustrate the physical interpretation of the formalism diagrammatically. By applying a self-consistency requirement, we derive the pair distributions...

  14. The carB gene encoding the large subunit of carbamoylphosphate synthetase from Lactococcus lactis is transcribed monocistronically

    DEFF Research Database (Denmark)

    Martinussen, Jan; Hammer, Karin

    1998-01-01

    The biosynthesis of carbamoylphosphate is catalysed by the heterodimeric enzyme carbamoylphosphate synthetase (CPSase). The genes encoding the two subunits in procaryotes are normally transcribed as an operon, whereas in Lactococcus lactis, the gene encoding the large subunit (carB) is shown...

  15. Two transgenic mouse models for β-subunit components of succinate-CoA ligase yielding pleiotropic metabolic alterations

    DEFF Research Database (Denmark)

    Kacso, Gergely; Ravasz, Dora; Doczi, Judit

    2016-01-01

    Succinate-CoA ligase (SUCL) is a heterodimer enzyme composed of Suclg1 α-subunit and a substrate-specific Sucla2 or Suclg2 β-subunit yielding ATP or GTP, respectively. In humans, the deficiency of this enzyme leads to encephalomyopathy with or without methylmalonyl aciduria, in addition to result...

  16. Dis3- and exosome subunit-responsive 3′ mRNA instability elements

    International Nuclear Information System (INIS)

    Kiss, Daniel L.; Hou, Dezhi; Gross, Robert H.; Andrulis, Erik D.

    2012-01-01

    Highlights: ► Successful use of a novel RNA-specific bioinformatic tool, RNA SCOPE. ► Identified novel 3′ UTR cis-acting element that destabilizes a reporter mRNA. ► Show exosome subunits are required for cis-acting element-mediated mRNA instability. ► Define precise sequence requirements of novel cis-acting element. ► Show that microarray-defined exosome subunit-regulated mRNAs have novel element. -- Abstract: Eukaryotic RNA turnover is regulated in part by the exosome, a nuclear and cytoplasmic complex of ribonucleases (RNases) and RNA-binding proteins. The major RNase of the complex is thought to be Dis3, a multi-functional 3′–5′ exoribonuclease and endoribonuclease. Although it is known that Dis3 and core exosome subunits are recruited to transcriptionally active genes and to messenger RNA (mRNA) substrates, this recruitment is thought to occur indirectly. We sought to discover cis-acting elements that recruit Dis3 or other exosome subunits. Using a bioinformatic tool called RNA SCOPE to screen the 3′ untranslated regions of up-regulated transcripts from our published Dis3 depletion-derived transcriptomic data set, we identified several motifs as candidate instability elements. Secondary screening using a luciferase reporter system revealed that one cassette—harboring four elements—destabilized the reporter transcript. RNAi-based depletion of Dis3, Rrp6, Rrp4, Rrp40, or Rrp46 diminished the efficacy of cassette-mediated destabilization. Truncation analysis of the cassette showed that two exosome subunit-sensitive elements (ESSEs) destabilized the reporter. Point-directed mutagenesis of ESSE abrogated the destabilization effect. An examination of the transcriptomic data from exosome subunit depletion-based microarrays revealed that mRNAs with ESSEs are found in every up-regulated mRNA data set but are underrepresented or missing from the down-regulated data sets. Taken together, our findings imply a potentially novel mechanism of m

  17. Overexpression of PP2A-C5 that encodes the catalytic subunit 5 of protein phosphatase 2A in Arabidopsis confers better root and shoot development under salt conditions

    Science.gov (United States)

    Protein phosphatase 2A (PP2A) is an enzyme consisting of three subunits: a scaffolding A subunit, a regulatory B subunit and a catalytic C subunit. PP2As were shown to play diverse roles in eukaryotes. In this study, the function of the Arabidopsis PP2A-C5 gene that encodes the catalytic subunit 5 o...

  18. [Three regions of Rpb10 mini-subunit of nuclear RNA polymerases are strictly conserved in all eukaryotes].

    Science.gov (United States)

    Shpakovskiĭ, G V; Lebedenko, E N

    1996-12-01

    The rpb10+ cDNA from the fission yeast Schizosaccharomyces pombe was cloned using two independent approaches (PCR and genetic suppression). The cloned cDNA encoded the Rpb10 subunit common for all three RNA polymerases. Comparison of the deduced amino acid sequence of the Sz. pombe Rbp10 subunit (71 amino acid residues) with those of the homologous subunits of RNA polymerases I, II, and III from Saccharomyces cerevisiae and Home sapiens revealed that heptapeptides RCFT/SCGK (residues 6-12), RYCCRRM (residues 43-49), and HVDLIEK (residues 53-59) were evolutionarily the most conserved structural motifs of these subunits. It is shown that the Rbp10 subunit from Sz. pombe can substitute its homolog (ABC10 beta) in the baker's yeast S. cerevisiae.

  19. Generation of recombinant antibodies to rat GABAA receptor subunits by affinity selection on synthetic peptides.

    Directory of Open Access Journals (Sweden)

    Sujatha P Koduvayur

    Full Text Available The abundance and physiological importance of GABAA receptors in the central nervous system make this neurotransmitter receptor an attractive target for localizing diagnostic and therapeutic biomolecules. GABAA receptors are expressed within the retina and mediate synaptic signaling at multiple stages of the visual process. To generate monoclonal affinity reagents that can specifically recognize GABAA receptor subunits, we screened two bacteriophage M13 libraries, which displayed human scFvs, by affinity selection with synthetic peptides predicted to correspond to extracellular regions of the rat α1 and β2 GABAA subunits. We isolated three anti-β2 and one anti-α1 subunit specific scFvs. Fluorescence polarization measurements revealed all four scFvs to have low micromolar affinities with their cognate peptide targets. The scFvs were capable of detecting fully folded GABAA receptors heterologously expressed by Xenopus laevis oocytes, while preserving ligand-gated channel activity. Moreover, A10, the anti-α1 subunit-specific scFv, was capable of detecting native GABAA receptors in the mouse retina, as observed by immunofluorescence staining. In order to improve their apparent affinity via avidity, we dimerized the A10 scFv by fusing it to the Fc portion of the IgG. The resulting scFv-Fc construct had a Kd of ∼26 nM, which corresponds to an approximately 135-fold improvement in binding, and a lower detection limit in dot blots, compared to the monomeric scFv. These results strongly support the use of peptides as targets for generating affinity reagents to membrane proteins and encourage investigation of molecular conjugates that use scFvs as anchoring components to localize reagents of interest at GABAA receptors of retina and other neural tissues, for studies of receptor activation and subunit structure.

  20. Structural model of the 50S subunit of E.Coli ribosomes from solution scattering

    Energy Technology Data Exchange (ETDEWEB)

    Svergun, D.I.; Koch, M.H.J. [Hamburg Outstation (Germany); Pedersen, J.S. [Riso National Laboratory, Roskilde (Denmark); Serdyuk, I.N. [Inst. of Protein Research, Moscow (Russian Federation)

    1994-12-31

    The application of new methods of small-angle scattering data interpretation to a contrast variation study of the 50S ribosomal subunit of Escherichia coli in solution is described. The X-ray data from contrast variation with sucrose are analyzed in terms of the basic scattering curves from the volume inaccessible to sucrose and from the regions inside this volume occupied mainly by RNA and by proteins. From these curves models of the shape of the 50S and its RNA-rich core are evaluated and positioned so that their difference produces a scattering curve which is in good agreement with the scattering from the protein moiety. Basing on this preliminary model, the X-ray and neutron contrast variation data of the 50S subunit in aqueous solutions are interpreted in the frame of the advanced two-phase model described by the shapes of the 50S subunit and its RNA-rich core taking into account density fluctuations inside the RNA and the protein moiety. The shape of the envelope of the 50S subunit and of the RNA-rich core are evaluated with a resolution of about 40A. The shape of the envelope is in good agreement with the models of the 50S subunit obtained from electron microscopy on isolated particles. The shape of the RNA-rich core correlates well with the model of the entire particle determined by the image reconstruction from ordered sheets indicating that the latter model which is based on the subjective contouring of density maps is heavily biased towards the RNA.