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Sample records for 4fe-4s clusters fa

  1. Electron transfer between [4Fe-4S] clusters

    NASA Astrophysics Data System (ADS)

    Voityuk, Alexander A.

    2010-07-01

    Iron-sulfur clusters [4Fe-4S] are major components in biological electron transport (ET). Using the DFT/B3LYP method, we calculate electronic coupling for low- and high-potential ET between [4Fe-4S] clusters and explore its sensitivity to structural parameters of the system and external electric field. As an example, we consider a model of the bacterial respiratory complex I and estimate the role of neighboring amino acids in facilitating the ET process between the clusters. Our results suggest that the superexchange mechanism rather than hole hopping should be operative in ET between [4Fe-4S] redox centers.

  2. The [4Fe-4S] cluster of quinolinate synthase from Escherichia coli: investigation of cluster ligands.

    PubMed

    Rousset, Carine; Fontecave, Marc; Ollagnier de Choudens, Sandrine

    2008-08-20

    Nicotinamide adenine dinucleotide (NAD) derives from quinolinic acid which is synthesized in Escherichia coli from l-aspartate and dihydroxyacetone phosphate through the concerted action of l-aspartate oxidase and the [4Fe-4S] quinolinate synthase (NadA). Here, we addressed the question of the identity of the cluster ligands. We performed in vivo complementation experiments as well as enzymatic, spectroscopic and structural in vitro studies using wild-type vs. Cys-to-Ala mutated NadA proteins. These studies reveal that only three cysteine residues, the conserved Cys113, Cys200 and Cys297, are ligands of the cluster. This result is in contrast to the previous proposal that pointed the three cysteines of the C(291)XXC(294)XXC(297) motif. Interestingly, we demonstrated that Cys291 and Cys294 form a disulfide bridge and are important for activity. PMID:18674537

  3. Dispensability of the [4Fe-4S] cluster in novel homologues of adenine glycosylase MutY.

    PubMed

    Trasviña-Arenas, Carlos H; Lopez-Castillo, Laura M; Sanchez-Sandoval, Eugenia; Brieba, Luis G

    2016-02-01

    7,8-Dihydro-8-deoxyguanine (8oG) is one of the most common oxidative lesions in DNA. DNA polymerases misincorporate an adenine across from this lesion. Thus, 8oG is a highly mutagenic lesion responsible for G:C→T:A transversions. MutY is an adenine glycosylase, part of the base excision repair pathway that removes adenines, when mispaired with 8oG or guanine. Its catalytic domain includes a [4Fe-4S] cluster motif coordinated by cysteinyl ligands. When this cluster is absent, MutY activity is depleted and several studies concluded that the [4Fe-4S] cluster motif is an indispensable component for DNA binding, substrate recognition and enzymatic activity. In the present study, we identified 46 MutY homologues that lack the canonical cysteinyl ligands, suggesting an absence of the [4Fe-4S] cluster. A phylogenetic analysis groups these novel MutYs into two different clades. One clade is exclusive of the order Lactobacillales and another clade has a mixed composition of anaerobic and microaerophilic bacteria and species from the protozoan genus Entamoeba. Structural modeling and sequence analysis suggests that the loss of the [4Fe-4S] cluster is compensated by a convergent solution in which bulky amino acids substitute the [4Fe-4S] cluster. We functionally characterized MutYs from Lactobacillus brevis and Entamoeba histolytica as representative members from each clade and found that both enzymes are active adenine glycosylases. Furthermore, chimeric glycosylases, in which the [4Fe-4S] cluster of Escherichia coli MutY is replaced by the corresponding amino acids of LbY and EhY, are also active. Our data indicates that the [4Fe-4S] cluster plays a structural role in MutYs and evidences the existence of alternative functional solutions in nature. PMID:26613369

  4. Protonation and Proton-Coupled Electron Transfer at S-Ligated [4Fe-4S] Clusters

    PubMed Central

    Morris, Wesley D.; Darcy, Julia W.; Mayer, James M.

    2015-01-01

    Biological [Fe-S] clusters are increasingly recognized to undergo proton-coupled electron transfer (PCET), but the site of protonation, mechanism, and role for PCET remains largely unknown. Here we explore this reactivity with synthetic model clusters. Protonation of the arylthiolate-ligated [4Fe-4S] cluster [Fe4S4(SAr)4]2- (1, SAr = S-2,4-6-(iPr)3C6H2) leads to thiol dissociation, reversibly forming [Fe4S4(SAr)3L]1- (2) + ArSH (L = solvent, and/or conjugate base). Solutions of 2 + ArSH react with the nitroxyl radical TEMPO to give [Fe4S4(SAr)4]1- (1ox) and TEMPOH. This reaction involves PCET coupled to thiolate association and may proceed via the unobserved protonated cluster [Fe4S4(SAr)3(HSAr)]1-(1-H). Similar reactions with this and related clusters proceed comparably. An understanding of the PCET thermochemistry of this cluster system has been developed, encompassing three different redox levels and two protonation states. PMID:25965413

  5. Protonation and Proton-Coupled Electron Transfer at S-Ligated [4Fe-4S] Clusters.

    PubMed

    Saouma, Caroline T; Morris, Wesley D; Darcy, Julia W; Mayer, James M

    2015-06-15

    Biological [Fe-S] clusters are increasingly recognized to undergo proton-coupled electron transfer (PCET), but the site of protonation, mechanism, and role for PCET remains largely unknown. Here we explore this reactivity with synthetic model clusters. Protonation of the arylthiolate-ligated [4Fe-4S] cluster [Fe4 S4 (SAr)4 ](2-) (1, SAr=S-2,4-6-(iPr)3 C6 H2 ) leads to thiol dissociation, reversibly forming [Fe4 S4 (SAr)3 L](1-) (2) and ArSH (L=solvent, and/or conjugate base). Solutions of 2+ArSH react with the nitroxyl radical TEMPO to give [Fe4 S4 (SAr)4 ](1-) (1ox ) and TEMPOH. This reaction involves PCET coupled to thiolate association and may proceed via the unobserved protonated cluster [Fe4 S4 (SAr)3 (HSAr)](1-) (1-H). Similar reactions with this and related clusters proceed comparably. An understanding of the PCET thermochemistry of this cluster system has been developed, encompassing three different redox levels and two protonation states. PMID:25965413

  6. Structure and Function of a Bacterial Microcompartment Shell Protein Engineered to Bind a [4Fe-4S] Cluster.

    PubMed

    Aussignargues, Clément; Pandelia, Maria-Eirini; Sutter, Markus; Plegaria, Jefferson S; Zarzycki, Jan; Turmo, Aiko; Huang, Jingcheng; Ducat, Daniel C; Hegg, Eric L; Gibney, Brian R; Kerfeld, Cheryl A

    2016-04-27

    Bacterial microcompartments (BMCs) are self-assembling organelles composed of a selectively permeable protein shell and encapsulated enzymes. They are considered promising templates for the engineering of designed bionanoreactors for biotechnology. In particular, encapsulation of oxidoreductive reactions requiring electron transfer between the lumen of the BMC and the cytosol relies on the ability to conduct electrons across the shell. We determined the crystal structure of a component protein of a synthetic BMC shell, which informed the rational design of a [4Fe-4S] cluster-binding site in its pore. We also solved the structure of the [4Fe-4S] cluster-bound, engineered protein to 1.8 Å resolution, providing the first structure of a BMC shell protein containing a metal center. The [4Fe-4S] cluster was characterized by optical and EPR spectroscopies; it has a reduction potential of -370 mV vs the standard hydrogen electrode (SHE) and is stable through redox cycling. This remarkable stability may be attributable to the hydrogen-bonding network provided by the main chain of the protein scaffold. The properties of the [4Fe-4S] cluster resemble those in low-potential bacterial ferredoxins, while its ligation to three cysteine residues is reminiscent of enzymes such as aconitase and radical S-adenosymethionine (SAM) enzymes. This engineered shell protein provides the foundation for conferring electron-transfer functionality to BMC shells. PMID:26704697

  7. Characterization of [4Fe-4S]-containing and cluster-free forms of Streptomyces WhiD

    PubMed Central

    Crack, Jason C.; den Hengst, Chris D.; Jakimowicz, Piotr; Subramanian, Sowmya; Johnson, Michael K.; Buttner, Mark J.; Thomson, Andrew J.; Le Brun, Nick E.

    2009-01-01

    WhiD, a member of the WhiB-like (Wbl) family of iron-sulfur proteins found exclusively within the actinomycetes, is required for the late stages of sporulation in Streptomyces coelicolor. Like all other Wbl proteins, WhiD has not so far been purified in a soluble form that contains a significant amount of cluster and characterization has relied on cluster-reconstituted protein. Thus, a major goal in Wbl research is to obtain and characterize native protein containing iron-sulfur clusters. Here we report the analysis of S. coelicolor WhiD purified anaerobically from E. coli as a soluble protein containing a single [4Fe-4S]2+ cluster ligated by four cysteines. Upon exposure to oxygen, spectral features associated with the [4Fe-4S] cluster were lost in a slow reaction that unusually yielded apo-WhiD directly without significant concentrations of cluster intermediates. This process was found to be highly pH dependent with an optimal stability observed between pH 7.0 and 8.0. Low molecular weight thiols, including a mycothiol analogue and thioredoxin, exerted a small but significant protective effect against WhiD cluster loss, an activity that could be of physiological importance. [4Fe-4S]2+ WhiD was found to react much more rapidly with superoxide than with either oxygen or hydrogen peroxide, which may also be of physiological significance. Loss of the [4Fe-4S] cluster to form apo-protein destabilized the protein fold significantly, but did not lead to complete unfolding. Finally, apo-WhiD exhibited negligible activity in an insulin-based disulfide reductase assay demonstrating that it does not function as a general protein disulfide reductase. PMID:19954209

  8. Crystal structure of 4-hydroxybutyryl-CoA dehydratase: radical catalysis involving a [4Fe-4S] cluster and flavin.

    PubMed

    Martins, Berta M; Dobbek, Holger; Cinkaya, Irfan; Buckel, Wolfgang; Messerschmidt, Albrecht

    2004-11-01

    Dehydratases catalyze the breakage of a carbon-oxygen bond leading to unsaturated products via the elimination of water. The 1.6-A resolution crystal structure of 4-hydroxybutyryl-CoA dehydratase from the gamma-aminobutyrate-fermenting Clostridium aminobutyricum represents a new class of dehydratases with an unprecedented active site architecture. A [4Fe-4S](2+) cluster, coordinated by three cysteine and one histidine residues, is located 7 A from the Re-side of a flavin adenine dinucleotide (FAD) moiety. The structure provides insight into the function of these ubiquitous prosthetic groups in the chemically nonfacile, radical-mediated dehydration of 4-hydroxybutyryl-CoA. The substrate can be bound between the [4Fe-4S](2+) cluster and the FAD with both cofactors contributing to its radical activation and catalytic conversion. Our results raise interesting questions regarding the mechanism of acyl-CoA dehydrogenases, which are involved in fatty acid oxidation, and address the divergent evolution of the ancestral common gene. PMID:15496473

  9. (4Fe-4S)-cluster-depleted Azotobacter vinelandii ferredoxin I: a new 3Fe iron-sulfur protein

    SciTech Connect

    Stephens, P.J.; Morgan, T.V.; Devlin, F.; Penner-Hahn, J.E.; Hodgson, K.O.; Scott, R.A.; Stout, C.D.; Burgess, B.K.

    1985-09-01

    Fe(CN)6T oxidation of the aerobically isolated 7Fe Azotobacter vinelandii ferredoxin I, (7Fe)FdI, is a degradative reaction. Destruction of the (4Fe-4S) cluster occurs first, followed by destruction of the (3Fe-3S) cluster. At a Fe(CN)6T /(7Fe)FdI concentration ratio of 20, the product is a mixture of apoprotein and protein containing only a (3Fe-3S) cluster, (3Fe)FdI. This protein mixture, after partial purification, has been characterized by absorption, CD, magnetic CD, and EPR and Fe x-ray absorption spectroscopies. EPR and magnetic CD spectra provide strong evidence that the (3Fe-3S) cluster in (3Fe)FdI is essentially identical in structure to that in (7Fe)FdI. Analysis of the extended x-ray absorption fine structure (EXAFS) of (3Fe)FdI finds Fe scattering at an average Fe...Fe distance of approx. =2.7 A. The structure of the oxidized (3Fe-3S) cluster in solutions of oxidized (3Fe)FdI, and, by extension, of oxidized (7Fe)FdI, is thus different from that obtained by x-ray crystallography on oxidized (7Fe)FdI. Possible interpretations of this result are discussed.

  10. A PAS domain with an oxygen labile [4Fe-4S](2+) cluster in the oxygen sensor kinase NreB of Staphylococcus carnosus.

    PubMed

    Müllner, Martin; Hammel, Oliver; Mienert, Bernd; Schlag, Steffen; Bill, Eckhard; Unden, Gottfried

    2008-12-30

    The cytoplasmic histidine sensor kinase NreB of Staphylococcus carnosus responds to O(2) and controls together with the response regulator NreC the expression of genes of nitrate/nitrite respiration. nreBC homologous genes were found in Staphylococcus strains and Bacillus clausii, and a modified form was found in some Lactobacillus strains. NreB contains a sensory domain with similarity to heme B binding PAS domains. Anaerobically prepared NreB of S. carnosus exhibited a (diamagnetic) [4Fe-4S](2+) cluster when assessed by Mossbauer spectroscopy. Upon reaction with air, the cluster was degraded with a half-life of approximately 2.5 min. No significant amounts of Mossbauer or EPR detectable intermediates were found during the decay, but magnetic Mossbauer spectra revealed formation of diamagnetic [2Fe-2S](2+) clusters. After extended exposure to air, NreB was devoid of a FeS cluster. Photoreduction with deazaflavin produced small amounts of [4Fe-4S](+), which were degraded subsequently. The magnetically perturbed Mossbauer spectrum of the [4Fe-4S](2+) cluster corroborated the S = 0 spin state and revealed uniform electric field gradient tensors of the iron sites, suggesting full delocalization of the valence electrons and binding of each of the Fe ions by four S ligands, including the ligand to the protein. Mutation of each of the four Cys residues inactivated NreB function in vivo in accordance with their role as ligands. [4Fe-4S](2+) cluster-containing NreB had high kinase activity. Exposure to air decreased the kinase activity and content of the [4Fe-4S](2+) cluster with similar half-lives. We conclude that the sensory domain of NreB represents a new type of PAS domain containing a [4Fe-4S](2+) cluster for sensing and function. PMID:19102705

  11. AcsF Catalyzes the ATP-dependent Insertion of Nickel into the Ni,Ni-[4Fe4S] Cluster of Acetyl-CoA Synthase.

    PubMed

    Gregg, Christina M; Goetzl, Sebastian; Jeoung, Jae-Hun; Dobbek, Holger

    2016-08-26

    Acetyl-CoA synthase (ACS) catalyzes the reversible condensation of CO, CoA, and a methyl-cation to form acetyl-CoA at a unique Ni,Ni-[4Fe4S] cluster (the A-cluster). However, it was unknown which proteins support the assembly of the A-cluster. We analyzed the product of a gene from the cluster containing the ACS gene, cooC2 from Carboxydothermus hydrogenoformans, named AcsFCh, and showed that it acts as a maturation factor of ACS. AcsFCh and inactive ACS form a stable 2:1 complex that binds two nickel ions with higher affinity than the individual components. The nickel-bound ACS-AcsFCh complex remains inactive until MgATP is added, thereby converting inactive to active ACS. AcsFCh is a MinD-type ATPase and belongs to the CooC protein family, which can be divided into homologous subgroups. We propose that proteins of one subgroup are responsible for assembling the Ni,Ni-[4Fe4S] cluster of ACS, whereas proteins of a second subgroup mature the [Ni4Fe4S] cluster of carbon monoxide dehydrogenases. PMID:27382049

  12. The conserved protein Dre2 uses essential [2Fe-2S] and [4Fe-4S] clusters for its function in cytosolic iron-sulfur protein assembly.

    PubMed

    Netz, Daili J A; Genau, Heide M; Weiler, Benjamin D; Bill, Eckhard; Pierik, Antonio J; Lill, Roland

    2016-07-15

    The cytosolic iron-sulfur (Fe-S) protein assembly (CIA) machinery comprises 11 essential components and matures Fe-S proteins involved in translation and genome maintenance. Maturation is initiated by the electron transfer chain NADPH-diflavin reductase Tah18-Fe-S protein Dre2 that facilitates the de novo assembly of a [4Fe-4S] cluster on the scaffold complex Cfd1-Nbp35. Tah18-Dre2 also play a critical role in the assembly of the diferric tyrosyl radical cofactor of ribonucleotide reductase. Dre2 contains eight conserved cysteine residues as potential co-ordinating ligands for Fe-S clusters but their functional importance and the type of bound clusters is unclear. In the present study, we use a combination of mutagenesis, cell biological and biochemical as well as UV-visible, EPR and Mössbauer spectroscopic approaches to show that the yeast Dre2 cysteine residues Cys(252), Cys(263), Cys(266) and Cys(268) (motif I) bind a [2Fe-2S] cluster, whereas cysteine residues Cys(311), Cys(314), Cys(322) and Cys(325) (motif II) co-ordinate a [4Fe-4S] cluster. All of these residues with the exception of Cys(252) are essential for cell viability, cytosolic Fe-S protein activity and in vivo (55)Fe-S cluster incorporation. The N-terminal methyltransferase-like domain of Dre2 is important for proper Fe-S cluster assembly at motifs I and II, which occurs in an interdependent fashion. Our findings further resolve why recombinant Dre2 from Arabidopsis, Trypanosoma or humans has previously been isolated with a single [2Fe-2S] instead of native [2Fe-2S] plus [4Fe-4S] clusters. In the presence of oxygen, the motif I-bound [2Fe-2S] cluster is labile and the motif II-bound [4Fe-4S] cluster is readily converted into a [2Fe-2S] cluster. PMID:27166425

  13. NsrR from Streptomyces coelicolor Is a Nitric Oxide-sensing [4Fe-4S] Cluster Protein with a Specialized Regulatory Function*

    PubMed Central

    Crack, Jason C.; Munnoch, John; Dodd, Erin L.; Knowles, Felicity; Al Bassam, Mahmoud M.; Kamali, Saeed; Holland, Ashley A.; Cramer, Stephen P.; Hamilton, Chris J.; Johnson, Michael K.; Thomson, Andrew J.; Hutchings, Matthew I.; Le Brun, Nick E.

    2015-01-01

    The Rrf2 family transcription factor NsrR controls expression of genes in a wide range of bacteria in response to nitric oxide (NO). The precise form of the NO-sensing module of NsrR is the subject of controversy because NsrR proteins containing either [2Fe-2S] or [4Fe-4S] clusters have been observed previously. Optical, Mössbauer, resonance Raman spectroscopies and native mass spectrometry demonstrate that Streptomyces coelicolor NsrR (ScNsrR), previously reported to contain a [2Fe-2S] cluster, can be isolated containing a [4Fe-4S] cluster. ChIP-seq experiments indicated that the ScNsrR regulon is small, consisting of only hmpA1, hmpA2, and nsrR itself. The hmpA genes encode NO-detoxifying flavohemoglobins, indicating that ScNsrR has a specialized regulatory function focused on NO detoxification and is not a global regulator like some NsrR orthologues. EMSAs and DNase I footprinting showed that the [4Fe-4S] form of ScNsrR binds specifically and tightly to an 11-bp inverted repeat sequence in the promoter regions of the identified target genes and that DNA binding is abolished following reaction with NO. Resonance Raman data were consistent with cluster coordination by three Cys residues and one oxygen-containing residue, and analysis of ScNsrR variants suggested that highly conserved Glu-85 may be the fourth ligand. Finally, we demonstrate that some low molecular weight thiols, but importantly not physiologically relevant thiols, such as cysteine and an analogue of mycothiol, bind weakly to the [4Fe-4S] cluster, and exposure of this bound form to O2 results in cluster conversion to the [2Fe-2S] form, which does not bind to DNA. These data help to account for the observation of [2Fe-2S] forms of NsrR. PMID:25771538

  14. Mutational analysis of the [4Fe-4S]-cluster converting iron regulatory factor from its RNA-binding form to cytoplasmic aconitase.

    PubMed Central

    Hirling, H; Henderson, B R; Kühn, L C

    1994-01-01

    The control of cellular iron homeostasis involves the coordinate post-transcriptional regulation of ferritin mRNA translation and transferring receptor mRNA stability. These regulatory events are mediated by a soluble cytoplasmic protein, iron regulatory factor (IRF), which binds specifically to mRNA hairpin structures, termed iron-responsive elements (IREs), in the respective mRNAs. IRF is modulated by variations of cellular iron levels and exists as either an apo-protein or a [4Fe-4S]-cluster protein. The two conformations show distinct, mutually exclusive functions. High-affinity IRE binding is observed with the apo-form induced by iron deprivation, but is lost under high iron conditions when IRF is converted to the [4Fe-4S]-cluster form which shows cytoplasmic aconitase activity. Moreover, IRE binding is inactivated by the sulfhydryl-oxidizing agent diamide and fully activated in vitro by 2% 2-mercapto-ethanol, whereas alkylation of IRF inhibits IRE binding. In the present study, we analyzed each of the above features using site-directed mutants of recombinant human IRF. The results support the bifunctional nature of IRF. We conclude that cysteines 437, 503 and 506 anchor the [4Fe-4S]-cluster, and are essential to the aconitase activity. Mutagenesis changing any of the cysteines to serine leads to constitutive RNA binding in 0.02% 2-mercaptoethanol. Cysteine 437 is particularly critical to the RNA-protein interaction. The spontaneous or diamide-induced formation of disulfide bonds between cysteines 437 and 503 or 437 and 506, in apo-IRF, as well as its alkylation by N-ethylmaleimide, inhibit binding to the IRE.(ABSTRACT TRUNCATED AT 250 WORDS) Images PMID:7508861

  15. Two [4Fe-4S] clusters containing radical SAM enzyme SkfB catalyze thioether bond formation during the maturation of the sporulation killing factor.

    PubMed

    Flühe, Leif; Burghaus, Olaf; Wieckowski, Beata M; Giessen, Tobias W; Linne, Uwe; Marahiel, Mohamed A

    2013-01-23

    The sporulation killing factor (SKF) is a 26-residue ribosomally assembled and posttranslationally modified sactipeptide. It is produced by Bacillus subtilis 168 and plays a key role in its sporulation. Like all sactipeptides, SKF contains a thioether bond, which links the cysteine residue Cys4 with the α-carbon of the methionine residue Met12. In this study we demonstrate that this bond is generated by the two [4Fe-4S] clusters containing radical SAM enzyme SkfB, which is encoded in the skf operon. By mutational analysis of both cluster-binding sites, we were able to postulate a mechanism for thioether generation which is in agreement with that of AlbA. Furthermore, we were able to show that thioether bond formation is specific toward hydrophobic amino acids at the acceptor site. Additionally we demonstrate that generation of the thioether linkage is leader-peptide-dependent, suggesting that this reaction is the first step in SKF maturation. PMID:23282011

  16. Insights into eukaryotic DNA priming from the structure and functional interactions of the 4Fe-4S cluster domain of human DNA primase

    SciTech Connect

    Vaithiyalingam, Sivaraja; Warren, Eric M.; Eichman, Brandt F.; Chazin, Walter J.

    2010-10-19

    DNA replication requires priming of DNA templates by enzymes known as primases. Although DNA primase structures are available from archaea and bacteria, the mechanism of DNA priming in higher eukaryotes remains poorly understood in large part due to the absence of the structure of the unique, highly conserved C-terminal regulatory domain of the large subunit (p58C). Here, we present the structure of this domain determined to 1.7-{angstrom} resolution by X-ray crystallography. The p58C structure reveals a novel arrangement of an evolutionarily conserved 4Fe-4S cluster buried deeply within the protein core and is not similar to any known protein structure. Analysis of the binding of DNA to p58C by fluorescence anisotropy measurements revealed a strong preference for ss/dsDNA junction substrates. This approach was combined with site-directed mutagenesis to confirm that the binding of DNA occurs to a distinctively basic surface on p58C. A specific interaction of p58C with the C-terminal domain of the intermediate subunit of replication protein A (RPA32C) was identified and characterized by isothermal titration calorimetry and NMR. Restraints from NMR experiments were used to drive computational docking of the two domains and generate a model of the p58C-RPA32C complex. Together, our results explain functional defects in human DNA primase mutants and provide insights into primosome loading on RPA-coated ssDNA and regulation of primase activity.

  17. 4-Demethylwyosine Synthase from Pyrococcus abyssi Is a Radical-S-adenosyl-l-methionine Enzyme with an Additional [4Fe-4S]+2 Cluster That Interacts with the Pyruvate Co-substrate*

    PubMed Central

    Perche-Letuvée, Phanélie; Kathirvelu, Velavan; Berggren, Gustav; Clemancey, Martin; Latour, Jean-Marc; Maurel, Vincent; Douki, Thierry; Armengaud, Jean; Mulliez, Etienne; Fontecave, Marc; Garcia-Serres, Ricardo; Gambarelli, Serge; Atta, Mohamed

    2012-01-01

    Wybutosine and its derivatives are found in position 37 of tRNA encoding Phe in eukaryotes and archaea. They are believed to play a key role in the decoding function of the ribosome. The second step in the biosynthesis of wybutosine is catalyzed by TYW1 protein, which is a member of the well established class of metalloenzymes called “Radical-SAM.” These enzymes use a [4Fe-4S] cluster, chelated by three cysteines in a CX3CX2C motif, and S-adenosyl-l-methionine (SAM) to generate a 5′-deoxyadenosyl radical that initiates various chemically challenging reactions. Sequence analysis of TYW1 proteins revealed, in the N-terminal half of the enzyme beside the Radical-SAM cysteine triad, an additional highly conserved cysteine motif. In this study we show by combining analytical and spectroscopic methods including UV-visible absorption, Mössbauer, EPR, and HYSCORE spectroscopies that these additional cysteines are involved in the coordination of a second [4Fe-4S] cluster displaying a free coordination site that interacts with pyruvate, the second substrate of the reaction. The presence of two distinct iron-sulfur clusters on TYW1 is reminiscent of MiaB, another tRNA-modifying metalloenzyme whose active form was shown to bind two iron-sulfur clusters. A possible role for the second [4Fe-4S] cluster in the enzyme activity is discussed. PMID:23043105

  18. A Cysteine-Rich CCG Domain Contains a Novel [4Fe-4S] Cluster Binding Motif As Deduced From Studies With Subunit B of Heterodisulfide Reductase From Methanothermobacter Marburgensis

    SciTech Connect

    Hamann, N.; Mander, G.J.; Shokes, J.E.; Scott, R.A.; Bennati, M.; Hedderich, R.

    2009-06-01

    Heterodisulfide reductase (HDR) of methanogenic archaea with its active-site [4Fe-4S] cluster catalyzes the reversible reduction of the heterodisulfide (CoM-S-S-CoB) of the methanogenic coenzyme M (CoM-SH) and coenzyme B (CoB-SH). CoM-HDR, a mechanistic-based paramagnetic intermediate generated upon half-reaction of the oxidized enzyme with CoM-SH, is a novel type of [4Fe-4S]{sup 3+} cluster with CoM-SH as a ligand. Subunit HdrB of the Methanothermobacter marburgensis HdrABC holoenzyme contains two cysteine-rich sequence motifs (CX{sub 31-39}CCX{sub 35-36}CXXC), designated as CCG domain in the Pfam database and conserved in many proteins. Here we present experimental evidence that the C-terminal CCG domain of HdrB binds this unusual [4Fe-4S] cluster. HdrB was produced in Escherichia coli, and an iron-sulfur cluster was subsequently inserted by in vitro reconstitution. In the oxidized state the cluster without the substrate exhibited a rhombic EPR signal (g{sub zyx} = 2.015, 1.995, and 1.950) reminiscent of the CoM-HDR signal. {sup 57}Fe ENDOR spectroscopy revealed that this paramagnetic species is a [4Fe-4S] cluster with {sup 57}Fe hyperfine couplings very similar to that of CoM-HDR. CoM-{sup 33}SH resulted in a broadening of the EPR signal, and upon addition of CoM-SH the midpoint potential of the cluster was shifted to values observed for CoM-HDR, both indicating binding of CoM-SH to the cluster. Site-directed mutagenesis of all 12 cysteine residues in HdrB identified four cysteines of the C-terminal CCG domain as cluster ligands. Combined with the previous detection of CoM-HDR-like EPR signals in other CCG domain-containing proteins our data indicate a general role of the C-terminal CCG domain in coordination of this novel [4Fe-4S] cluster. In addition, Zn K-edge X-ray absorption spectroscopy identified an isolated Zn site with an S{sub 3}(O/N){sub 1} geometry in HdrB and the HDR holoenzyme. The N-terminal CCG domain is suggested to provide ligands to the Zn

  19. Two EPR-detectable [4Fe-4S] clusters, N2a and N2b, are bound to the NuoI (TYKY) subunit of NADH:ubiquinone oxidoreductase (Complex I) from Rhodobacter capsulatus.

    PubMed

    Chevallet, Mireille; Dupuis, Alain; Issartel, Jean-Paul; Lunardi, Joël; van Belzen, Ronald; Albracht, Simon P J

    2003-03-01

    NADH:ubiquinone oxidoreductases (Complex I) contain a subunit, TYKY in the bovine enzyme and NuoI in the enzyme from Rhodobacter capsulatus, which is assumed to bind two [4Fe-4S] clusters because it contains two sets of conserved cysteine motifs similar to those found in the 2[4Fe-4S] ferredoxins. It was recently shown that the TYKY subunit is not an ordinary 2[4Fe-4S] ferredoxin, but has a unique amino acid sequence, which is only found in NAD(P)H:quinone oxidoreductases and certain membrane-bound [NiFe]-hydrogenases expected to be involved in redox-linked proton translocation [FEBS Lett. 485 (2000) 1]. We have generated a set of R. capsulatus mutants in which five out of the eight conserved cysteine residues in NuoI were replaced by other amino acids. The resulting mutants fell into three categories with virtually no, intermediate or quite normal Complex I activities. EPR-spectroscopic analysis of the membranes of the C67S and C106S mutants, two mutants belonging to the second and third group, respectively, showed a specific 50% decrease of the EPR signal attributed to cluster N2. It is concluded that the NuoI (TYKY) subunit binds two clusters N2, called N2a and N2b, which exhibit very similar spectral features when analyzed by X-band EPR spectroscopy. PMID:12615348

  20. Dynamics of the [4Fe-4S] Cluster in Pyrococcus furiosus D14C Ferredoxin via Nuclear Resonance Vibrational and Resonance Raman Spectroscopies, Force Field Simulations, and Density Functional Theory Calculations

    PubMed Central

    Mitra, Devrani; Pelmenschikov, Vladimir; Guo, Yisong; Case, David A.; Wang, Hongxin; Dong, Weibing; Tan, Ming-Liang; Ichiye, Toshiko; Jenney, Francis E.; Adams, Michael W. W.; Yoda, Yoshitaka; Zhao, Jiyong; Cramer, Stephen P.

    2011-01-01

    We have used 57Fe nuclear resonance vibrational spectroscopy (NRVS) to study oxidized and reduced forms of the [4Fe-4S] cluster in the D14C variant ferredoxin from Pyrococcus furiosus (Pf D14C Fd). To assist the normal mode assignments, we recorded the NRVS of D14C ferredoxin samples with 36S substituted into the [4Fe-4S] cluster bridging sulfide positions, and a model compound without ligand side chains: (Ph4P)2[Fe4S4Cl4]. Several distinct regions of NRVS intensity are identified, ranging from `protein' and torsional modes below 100 cm−1, through bending and breathing modes near 150 cm−1, to strong bands from Fe-S stretching modes between 250 cm−1 and ~400 cm−1. The oxidized ferredoxin samples were also investigated by resonance Raman (RR) spectroscopy. We found good agreement between NRVS and RR frequencies, but because of different selection rules, the intensities vary dramatically between the two types of spectra. The 57Fe partial vibrational densities of states (PVDOS) for the oxidized samples were interpreted by normal mode analysis with optimization of Urey-Bradley force fields for local models of the [4Fe-4S] clusters. Full protein model calculations were also conducted using a supplemented CHARMM force field, and these calculations revealed low frequency modes that may be relevant to electron transfer with Pf Fd partners. Density functional theory (DFT) calculations complemented these empirical analyses, and DFT was used to estimate the reorganization energy associated with the [Fe4S4]2+/1+ redox cycle. Overall, the NRVS technique demonstrates great promise for the observation and quantitative interpretation of the dynamical properties of Fe-S proteins. PMID:21500788

  1. Evidence That Phosphorylation of Iron Regulatory Protein 1 at Serine 138 Destabilizes the [4Fe-4S] Cluster in Cytosolic Aconitase by Enhancing 4Fe-3Fe Cycling*S⃞

    PubMed Central

    Deck, Kathryn M.; Vasanthakumar, Aparna; Anderson, Sheila A.; Goforth, Jeremy B.; Kennedy, M. Claire; Antholine, William E.; Eisenstein, Richard S.

    2009-01-01

    Iron-sulfur cluster-dependent interconversion of iron regulatory protein 1 (IRP1) between its RNA binding and cytosolic aconitase (c-acon) forms controls vertebrate iron homeostasis. Cluster removal from c-acon is thought to include oxidative demetallation as a required step, but little else is understood about the process of conversion to IRP1. In comparison with c-aconWT, Ser138 phosphomimetic mutants of c-acon contain an unstable [4Fe-4S] cluster and were used as tools to further define the pathway(s) of iron-sulfur cluster disassembly. Under anaerobic conditions cluster insertion into purified IRP1S138E and cluster loss on treatment with NO regulated aconitase and RNA binding activity over a similar range as observed for IRP1WT. However, activation of RNA binding of c-aconS138E was an order of magnitude more sensitive to NO than for c-aconWT. Consistent with this, an altered set point between RNA-binding and aconitase forms was observed for IRP1S138E when expressed in HEK cells. Active c-aconS138E could only accumulate under hypoxic conditions, suggesting enhanced cluster disassembly in normoxia. Cluster disassembly mechanisms were further probed by determining the impact of iron chelation on acon activity. Unexpectedly EDTA rapidly inhibited c-aconS138E activity without affecting c-aconWT. Additional chelator experiments suggested that cluster loss can be initiated in c-aconS138E through a spontaneous nonoxidative demetallation process. Taken together, our results support a model wherein Ser138 phosphorylation sensitizes IRP1/c-acon to decreased iron availability by allowing the [4Fe-4S]2+ cluster to cycle with [3Fe-4S]0 in the absence of cluster perturbants, indicating that regulation can be initiated merely by changes in iron availability. PMID:19269970

  2. Isoprenoid Biosynthesis in Pathogenic Bacteria: Nuclear Resonance Vibrational Spectroscopy Provides Insight into the Unusual [4Fe-4S] Cluster of the E. coli LytB/IspH Protein.

    PubMed

    Faus, Isabelle; Reinhard, Annegret; Rackwitz, Sergej; Wolny, Juliusz A; Schlage, Kai; Wille, Hans-Christian; Chumakov, Aleksandr; Krasutsky, Sergiy; Chaignon, Philippe; Poulter, C Dale; Seemann, Myriam; Schünemann, Volker

    2015-10-19

    The LytB/IspH protein catalyzes the last step of the methylerythritol phosphate (MEP) pathway which is used for the biosynthesis of essential terpenoids in most pathogenic bacteria. Therefore, the MEP pathway is a target for the development of new antimicrobial agents as it is essential for microorganisms, yet absent in humans. Substrate-free LytB has a special [4Fe-4S](2+) cluster with a yet unsolved structure. This motivated us to use synchrotron-based nuclear resonance vibrational spectroscopy (NRVS) in combination with quantum chemical-molecular mechanical (QM/MM) calculations to gain more insight into the structure of substrate-free LytB. The apical iron atom of the [4Fe-4S](2+) is clearly linked to three water molecules. We additionally present NRVS data of LytB bound to its natural substrate, (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate (HMBPP) and to the inhibitors (E)-4-amino-3-methylbut-2-en-1-yl diphosphate and (E)-4-mercapto-3-methylbut-2-en-1-yl diphosphate. PMID:26118554

  3. Electrochemical Resolution of the [4Fe-4S] Centers of the AdoMet Radical Enzyme BtrN: Evidence of Proton Coupling and an Unusual, Low-Potential Auxiliary Cluster.

    PubMed

    Maiocco, Stephanie J; Grove, Tyler L; Booker, Squire J; Elliott, Sean J

    2015-07-15

    The S-adenosylmethionine (AdoMet) radical superfamily of enzymes includes over 113,500 unique members, each of which contains one indispensable iron-sulfur (FeS) cluster that is required to generate a 5'-deoxyadenosyl 5'-radical intermediate during catalysis. Enzymes within several subgroups of the superfamily, however, have been found to contain one or more additional FeS clusters. While these additional clusters are absolutely essential for enzyme activity, their exact roles in the function and/or mechanism of action of many of the enzymes are at best speculative, indicating a need to develop methods to characterize and study these clusters in more detail. Here, BtrN, an AdoMet radical dehydrogenase that catalyzes the two-electron oxidation of 2-deoxy-scyllo-inosamine to amino-dideoxy-scyllo-inosose, an intermediate in the biosynthesis of 2-deoxystreptamine antibiotics, is examined through direct electrochemistry, where the potential of both its AdoMet radical and auxiliary [4Fe-4S] clusters can be measured simultaneously. We find that the AdoMet radical cluster exhibits a midpoint potential of -510 mV, while the auxiliary cluster exhibits a midpoint potential of -765 mV, to our knowledge the lowest [4Fe-4S](2+/+) potential to be determined to date. The impact of AdoMet binding and the pH dependence of catalysis are also quantitatively observed. These data show that direct electrochemical methods can be used to further elucidate the chemistry of the burgeoning AdoMet radical superfamily in the future. PMID:26088836

  4. sup 17 O, sup 1 H, and sup 2 H electron nuclear double resonance characterization of solvent, substrate, and inhibitor binding to the (4Fe-4S) sup + cluster of aconitase

    SciTech Connect

    Werst, M.M.; Hoffman, B.M. ); Kennedy, M.C.; Beinert, H. )

    1990-11-01

    {sup 17}O electron nuclear double resonance (ENDOR) studies at X-band (9-GHz) and Q-band (35-GHz) microwave frequencies reveal that the (4Fe-4S){sup {plus}} cluster of substrate-free aconitase (citrate (isocitrate) hydro-lyase, EC 4.2.1.3) binds solvent, H{sub x}O (x = 1,2). Previous {sup 17}O ENDOR studies had disclosed that H{sub x}{sup 17}O binds to the enzyme-substrate complex and also to complexes of enzyme with the substrate analogues trans-aconitate and nitroisocitrate (1-hydroxy-2-nitro-1,3-propanedicarboxylate). The authors have used {sup 1}H and {sup 2}H ENDOR to characterize these solvent species. The authors propose that the fourth ligand of Fe{sub a} in substrate-free enzyme is a hydroxyl ion from the solvent; upon binding of substrate or substrate analogues at this Fe{sub a} site, the solvent species becomes protonated to form a water molecule. Previous {sup 17}O and {sup 13}C ENDOR studies showed that only a single carboxyl, at C-2 of the propane backbone of cis-aconitate or at C-1 of the inhibitor nitroisocitrate, coordinates to the cluster. Together, these results imply that enzyme-catalyzed interconversion of citrate and isocitrate does not involve displacement of an endogenous fourth ligand, but rather addition of the anionic carboxylate ligand and a change in protonation state of a solvent species bound to Fe{sub a}. The authors further report the {sup 17}O hyperfine tensor parameters of the C-2 carboxyl oxygen of substrate bound to the cluster as determined by the field dependence of the {sup 17}O ENDOR signals. {sup 17}O ENDOR studies also show that the carboxyl group of the inhibitor trans-aconitate binds similarly to that off substrate.

  5. Atomic structure of the DNA repair [4Fe-4S] enzyme endonuclease III.

    PubMed

    Kuo, C F; McRee, D E; Fisher, C L; O'Handley, S F; Cunningham, R P; Tainer, J A

    1992-10-16

    The crystal structure of the DNA repair enzyme endonuclease III, which recognizes and cleaves DNA at damaged bases, has been solved to 2.0 angstrom resolution with an R factor of 0.185. This iron-sulfur [4Fe-4S] enzyme is elongated and bilobal with a deep cleft separating two similarly sized domains: a novel, sequence-continuous, six-helix domain (residues 22 to 132) and a Greek-key, four-helix domain formed by the amino-terminal and three carboxyl-terminal helices (residues 1 to 21 and 133 to 211) together with the [4Fe-4S] cluster. The cluster is bound entirely within the carboxyl-terminal loop with a ligation pattern (Cys-X6-Cys-X2-Cys-X5-Cys) distinct from all other known [4Fe-4S] proteins. Sequence conservation and the positive electrostatic potential of conserved regions identify a surface suitable for binding duplex B-DNA across the long axis of the enzyme, matching a 46 angstrom length of protected DNA. The primary role of the [4Fe-4S] cluster appears to involve positioning conserved basic residues for interaction with the DNA phosphate backbone. The crystallographically identified inhibitor binding region, which recognizes the damaged base thymine glycol, is a seven-residue beta-hairpin (residues 113 to 119). Location and side chain orientation at the base of the inhibitor binding site implicate Glu112 in the N-glycosylase mechanism and Lys120 in the beta-elimination mechanism. Overall, the structure reveals an unusual fold and a new biological function for [4Fe-4S] clusters and provides a structural basis for studying recognition of damaged DNA and the N-glycosylase and apurinic/apyrimidinic-lyase mechanisms. PMID:1411536

  6. Structure of the non-redox-active tungsten/[4Fe:4S] enzyme acetylene hydratase.

    PubMed

    Seiffert, Grazyna B; Ullmann, G Matthias; Messerschmidt, Albrecht; Schink, Bernhard; Kroneck, Peter M H; Einsle, Oliver

    2007-02-27

    The tungsten-iron-sulfur enzyme acetylene hydratase stands out from its class because it catalyzes a nonredox reaction, the hydration of acetylene to acetaldehyde. Sequence comparisons group the protein into the dimethyl sulfoxide reductase family, and it contains a bis-molybdopterin guanine dinucleotide-ligated tungsten atom and a cubane-type [4Fe:4S] cluster. The crystal structure of acetylene hydratase at 1.26 A now shows that the tungsten center binds a water molecule that is activated by an adjacent aspartate residue, enabling it to attack acetylene bound in a distinct, hydrophobic pocket. This mechanism requires a strong shift of pK(a) of the aspartate, caused by a nearby low-potential [4Fe:4S] cluster. To access this previously unrecognized W-Asp active site, the protein evolved a new substrate channel distant from where it is found in other molybdenum and tungsten enzymes. PMID:17360611

  7. Structure and Function of Four Classes of the 4Fe-4S Protein, IspH.

    PubMed

    Rao, Guodong; Oldfield, Eric

    2016-07-26

    IspH, (E)-1-hydroxy-2-methyl-but-2-enyl 4-diphosphate reductase, is an essential enzyme in isoprenoid biosynthesis and an important drug/herbicide target. Using X-ray crystallographic, bioinformatics, mutagenesis/kinetics/stability, and electron paramagnetic resonance (EPR) results, we show that organisms from different environments ultilize one of four main IspH classes. The classes are based on the arrangement of the aromatic residues near the 4Fe-4S cluster and the presence or absence of N- and C-terminal extensions. Class A enzymes are found primarily in anaerobic and microaerophilic bacteria. Class B enzymes are found in aerobic bacteria. Class C enzymes are found in cyanobacteria and plants. Class D enzymes are found in apicomplexan parasites. Using mutagenesis, we show that the cluster-associated aromatic groups in class A and class B IspHs enhance cluster oxidative stability. Y198A, F302A, and a C-terminal truncation mutant of the class B (Escherichia coli) IspH have catalytic activity lower than that of the wild-type protein when using methyl viologen as the electron donor, but higher activity with dithionite as the electron donor, due to ready access of the small reductant to the cluster, consistent with their increased oxygen and H2O2 sensitivity. F302A has the largest effect on the reaction rates, and EPR studies indicate this residue affects Fe-S cluster structure. Similar effects on cluster stability are seen with class A (F14A and Y98A) mutants; however, effects on ET rates are smaller, and there are no differences between the EPR spectra of mutant and wild-type proteins. Overall, the results are of general interest because they show, for the first time, that there are multiple IspH classes that have evolved to allow organisms to survive in diverse oxidative-stress environments. PMID:27357244

  8. Differentiated, Promoter-specific Response of [4Fe-4S] NsrR DNA Binding to Reaction with Nitric Oxide*

    PubMed Central

    Crack, Jason C.; Svistunenko, Dimitri A.; Munnoch, John; Thomson, Andrew J.; Hutchings, Matthew I.; Le Brun, Nick E.

    2016-01-01

    NsrR is an iron-sulfur cluster protein that regulates the nitric oxide (NO) stress response of many bacteria. NsrR from Streptomyces coelicolor regulates its own expression and that of only two other genes, hmpA1 and hmpA2, which encode HmpA enzymes predicted to detoxify NO. NsrR binds promoter DNA with high affinity only when coordinating a [4Fe-4S] cluster. Here we show that reaction of [4Fe-4S] NsrR with NO affects DNA binding differently depending on the gene promoter. Binding to the hmpA2 promoter was abolished at ∼2 NO per cluster, although for the hmpA1 and nsrR promoters, ∼4 and ∼8 NO molecules, respectively, were required to abolish DNA binding. Spectroscopic and kinetic studies of the NO reaction revealed a rapid, multi-phase, non-concerted process involving up to 8–10 NO molecules per cluster, leading to the formation of several iron-nitrosyl species. A distinct intermediate was observed at ∼2 NO per cluster, along with two further intermediates at ∼4 and ∼6 NO. The NsrR nitrosylation reaction was not significantly affected by DNA binding. These results show that NsrR regulates different promoters in response to different concentrations of NO. Spectroscopic evidence indicates that this is achieved by different NO-FeS complexes. PMID:26887943

  9. Differentiated, Promoter-specific Response of [4Fe-4S] NsrR DNA Binding to Reaction with Nitric Oxide.

    PubMed

    Crack, Jason C; Svistunenko, Dimitri A; Munnoch, John; Thomson, Andrew J; Hutchings, Matthew I; Le Brun, Nick E

    2016-04-15

    NsrR is an iron-sulfur cluster protein that regulates the nitric oxide (NO) stress response of many bacteria. NsrR from Streptomyces coelicolor regulates its own expression and that of only two other genes, hmpA1 and hmpA2, which encode HmpA enzymes predicted to detoxify NO. NsrR binds promoter DNA with high affinity only when coordinating a [4Fe-4S] cluster. Here we show that reaction of [4Fe-4S] NsrR with NO affects DNA binding differently depending on the gene promoter. Binding to the hmpA2 promoter was abolished at ∼2 NO per cluster, although for the hmpA1 and nsrR promoters, ∼4 and ∼8 NO molecules, respectively, were required to abolish DNA binding. Spectroscopic and kinetic studies of the NO reaction revealed a rapid, multi-phase, non-concerted process involving up to 8-10 NO molecules per cluster, leading to the formation of several iron-nitrosyl species. A distinct intermediate was observed at ∼2 NO per cluster, along with two further intermediates at ∼4 and ∼6 NO. The NsrR nitrosylation reaction was not significantly affected by DNA binding. These results show that NsrR regulates different promoters in response to different concentrations of NO. Spectroscopic evidence indicates that this is achieved by different NO-FeS complexes. PMID:26887943

  10. Cloning, expression, and molecular characterization of the gene encoding an extremely thermostable [4Fe-4S] ferredoxin from the hyperthermophilic archaeon Pyrococcus furiosus.

    PubMed Central

    Heltzel, A; Smith, E T; Zhou, Z H; Blamey, J M; Adams, M W

    1994-01-01

    The gene for ferredoxin from the hyperthermophilic archaeon Pyrococcus furiosus was cloned, sequenced, and expressed in Escherichia coli. The coding region confirmed the determined amino acid sequence. Putative archaeon-type transcriptional regulatory elements were identified. The fdxA gene appears to be an independent transcriptional unit. Recombinant ferredoxin was indistinguishable from the protein purified from P. furiosus in its thermal stability and in the potentiometric and spectroscopic properties of its [4Fe-4S] cluster. PMID:8045914

  11. The reducing component BoxA of benzoyl-coenzyme A epoxidase from Azoarcus evansii is a [4Fe-4S] protein.

    PubMed

    Rather, Liv J; Bill, Eckhard; Ismail, Wael; Fuchs, Georg

    2011-12-01

    BoxA is the reductase component of the benzoyl-coenzyme A (CoA) oxidizing epoxidase enzyme system BoxAB. The enzyme catalyzes the key step of an hitherto unknown aerobic, CoA-dependent pathway of benzoate metabolism, which is the epoxidation of benzoyl-CoA to the non-aromatic 2,3-epoxybenzoyl-CoA. The function of BoxA is the transfer of two electrons from NADPH to the epoxidase component BoxB. We could show recently that BoxB is a diiron enzyme, whereas here we demonstrate that BoxA harbors an FAD and two [4Fe-4S] clusters per protein monomer. The characterization of BoxA was hampered by severe oxygen sensitivity; the cubane [4Fe-4S] clusters degrade already with traces of oxygen. Interestingly, the adventitiously formed [3Fe-4S] centers could be reconstituted in vitro by adding Fe(II) and sulfide to retrieve the native cubane centers. BoxA is the first example of a reductase of this type that has an FAD and two bacterial ferredoxin-type [4Fe-4S] clusters. In other cases within the catalytically versatile family of diiron enzymes, the related reductases have plant-type ferredoxin or Rieske-type [2Fe-2S] centers only. PMID:21672639

  12. Crystallographic analysis of the intact metal centres [3Fe-4S](1+/0) and [4Fe-4S](2+/1+) in a Zn(2+) -containing ferredoxin.

    PubMed

    Frazão, Carlos; Aragão, David; Coelho, Ricardo; Leal, Sónia S; Gomes, Cláudio M; Teixeira, Miguel; Carrondo, Maria Arménia

    2008-03-01

    Detailed structural models of di-cluster seven-iron ferredoxins constitute a valuable resource for folding and stability studies relating the metal cofactors' role in protein stability. The here reported, hemihedric twinned crystal structure at 2.0 A resolution from Acidianus ambivalens ferredoxin, shows an integral 103 residues, physiologically relevant native form composed by a N-terminal extension comprising a His/Asp Zn(2+) site and the ferredoxin (betaalphabeta)(2) core, which harbours intact clusters I and II, a [3Fe-4S](1+/0) and a [4Fe-4S](2+/1+) centres. This is in contrast with the previously available ferredoxin structure from Sulfolofus tokodai, which was obtained from an artificial oxidative conversion with two [3Fe-4S](1+/0) centres and poor definition around cluster II. PMID:18258200

  13. Inhibition of the 4Fe-4S Proteins IspG and IspH: an EPR, ENDOR and HYSCORE Investigation

    PubMed Central

    Guerra, Francisco; Wang, Ke; Li, Jikun; Wang, Weixue; Liu, Yi-Liang; Amin, Shivani

    2014-01-01

    IspG and IspH are proteins that are involved in isoprenoid biosynthesis in most bacteria as well as in malaria parasites and are important drug targets. They contain cubane-type 4Fe-4S clusters that are involved in unusual 2H+/2e− reductions. Here, we report the results of electron paramagnetic resonance spectroscopic investigations of the binding of amino- and thiolo-HMBPP (HMBPP=E-1-hydroxy-2-methyl-but-2-enyl 4-diphosphate) IspH substrate-analog inhibitors to both proteins, as well as the binding of HMBPP and an acetylene diphosphate inhibitor, to IspG. The results show that amino-HMBPP binds to reduced IspH by Fe-C π-bonding with the olefinic carbons interacting with the unique 4th Fe in the 4Fe-4S cluster, quite different to the direct Fe-N ligation seen with the oxidized protein. No such π-complex is observed when amino-HMBPP binds to reduced IspG. No EPR signal is observed with IspH in the presence of dithionite and thiolo-HMBPP, suggesting that the 4Fe-4S cluster is not reduced, consistent with the presence of a 420 nm feature in the absorption spectrum (characteristic of an oxidized cluster). However, with IspG, the EPR spectrum in the presence of dithionite and thiolo-HMBPP is very similar to that seen with HMBPP. The binding of HMBPP to IspG was studied using hyperfine sublevel correlation spectroscopy with 17O and 13C labeled samples: the results rule out direct Fe-O bonding and indicate π-bonding. Finally, the binding to IspG of a potent acetylene diphosphate inhibitor was studied by using electron-nuclear double resonance spectroscopy with 13C labeled ligands: the large hyperfine couplings indicate strong Fe-C π-bonding with the acetylenic group. These results illustrate a remarkable diversity in binding behavior for HMBPP-analog inhibitors, opening up new routes to inhibitor design of interest in the context of anti-bacterial and anti-malarial drug discovery, as well as “cubane-type” metallo-biochemistry, in general. PMID:24999381

  14. Heterologous overproduction of 2[4Fe4S]- and [2Fe2S]-type clostridial ferredoxins and [2Fe2S]-type agrobacterial ferredoxin.

    PubMed

    Huang, Haiyan; Hu, Liejie; Yu, Wenjun; Li, Huili; Tao, Fei; Xie, Huijun; Wang, Shuning

    2016-05-01

    Ferredoxins are small, acidic proteins containing iron-sulfur clusters that are widespread in living organisms. They play key roles as electron carriers in various metabolic processes, including respiration, photosynthesis, fermentation, nitrogen fixation, carbon dioxide fixation, and hydrogen production. However, only several kinds of ferredoxins are commercially available now, greatly limiting the investigation of ferredoxin-related enzymes and metabolic processes. Here we describe the heterologous overproduction of 2[4Fe4S]- and [2Fe2S]-type clostridial ferredoxins and [2Fe2S]-type agrobacterial ferredoxin. Adding extra iron and sulfur sources to the medium in combination with using Escherichia coli C41(DE3) harboring pCodonplus and pRKISC plasmids as host greatly enhanced iron-sulfur cluster synthesis in the three ferredoxins. After induction for 12 h in terrific broth and purification by affinity chromatography and anion exchange chromatography, approximately 3.4 mg of streptavidin (Strep)-tagged and 3.7 mg of polyhistidine (His)-tagged clostridial 2[4Fe4S] ferredoxins were obtained from 1 l of culture. Excitingly, after induction for 24 h in terrific broth, around 40 mg of His-tagged clostridial [2Fe2S] and 23 mg of His-tagged agrobacterial [2Fe2S] ferredoxins were purified from 1 l of culture. The recombinant ferredoxins apparently exhibited identical properties and physiological function to native ferredoxins. No negative impact of two different affinity tags on ferredoxin activity was found. In conclusion, we successfully developed a convenient method for heterologous overproduction of the three kinds of ferredoxins with satisfactory yields and activities, which would be very helpful for the ferredoxin-related researches. PMID:26748213

  15. In Vitro Characterization of AtsB, a Radical SAM Formylglycine-Generating Enzyme That Contains Three [4Fe-4S] Clusters†

    PubMed Central

    Grove, Tyler L.; Lee, Kyung-Hoon; St. Clair, Jennifer; Krebs, Carsten; Booker, Squire J.

    2009-01-01

    Sulfatases catalyze the cleavage of a variety of cellular sulfate esters via a novel mechanism that requires the action of a protein-derived formylglycine cofactor. Formation of the cofactor is catalyzed by an accessory protein and involves the two-electron oxidation of a specific cysteinyl or seryl residue on the relevant sulfatase. Although some sulfatases undergo maturation via mechanisms in which oxygen serves as an electron acceptor, AtsB, the maturase from Klebsiella pneumoniae, catalyzes the oxidation of Ser72 on AtsA, its cognate sulfatase, via an oxygen-independent mechanism. Moreover, it does not make use of pyridine or flavin nucleotide cofactors as direct electron acceptors. In fact, AtsB has been shown to be a member of the radical S-adenosylmethionine superfamily of proteins, suggesting that it catalyzes this oxidation via an intermediate 5′-deoxyadenosyl 5′-radical that is generated by a reductive cleavage of S-adenosyl-L-methionine. In contrast to AtsA, very little in vitro characterization of AtsB has been conducted. Herein we show that coexpression of the K. pneumoniae atsB gene with a plasmid that encodes genes that are known to be involved in iron–sulfur cluster biosynthesis yields soluble protein that can be characterized in vitro. The as-isolated protein contained 8.7 ± 0.4 irons and 12.2 ± 2.6 sulfides per polypeptide, which existed almost entirely in the [4Fe-4S]2+ configuration, as determined by Mössbauer spectroscopy, suggesting that it contained at least two of these clusters per polypeptide. Reconstitution of the as-isolated protein with additional iron and sulfide indicated the presence of 12.3 ± 0.2 irons and 9.9 ± 0.4 sulfides per polypeptide. Subsequent characterization of the reconstituted protein by Mössbauer spectroscopy indicated the presence of only [4Fe-4S] clusters, suggesting that reconstituted AtsB contains three per polypeptide. Consistent with this stoichiometry, an as-isolated AtsB triple variant containing

  16. Structural principles for computational and de novo design of 4Fe-4S metalloproteins.

    PubMed

    Nanda, Vikas; Senn, Stefan; Pike, Douglas H; Rodriguez-Granillo, Agustina; Hansen, Will A; Khare, Sagar D; Noy, Dror

    2016-05-01

    Iron-sulfur centers in metalloproteins can access multiple oxidation states over a broad range of potentials, allowing them to participate in a variety of electron transfer reactions and serving as catalysts for high-energy redox processes. The nitrogenase FeMoCO cluster converts di-nitrogen to ammonia in an eight-electron transfer step. The 2(Fe4S4) containing bacterial ferredoxin is an evolutionarily ancient metalloprotein fold and is thought to be a primordial progenitor of extant oxidoreductases. Controlling chemical transformations mediated by iron-sulfur centers such as nitrogen fixation, hydrogen production as well as electron transfer reactions involved in photosynthesis are of tremendous importance for sustainable chemistry and energy production initiatives. As such, there is significant interest in the design of iron-sulfur proteins as minimal models to gain fundamental understanding of complex natural systems and as lead-molecules for industrial and energy applications. Herein, we discuss salient structural characteristics of natural iron-sulfur proteins and how they guide principles for design. Model structures of past designs are analyzed in the context of these principles and potential directions for enhanced designs are presented, and new areas of iron-sulfur protein design are proposed. This article is part of a Special issue entitled Biodesign for Bioenergetics - the design and engineering of electronic transfer cofactors, protein networks, edited by Ronald L. Koder and J.L Ross Anderson. PMID:26449207

  17. Electron spin-lattice relaxation of the (4Fe-4S) ferredoxin from B. stearothermophilus. Comparison with other iron proteins

    NASA Astrophysics Data System (ADS)

    Bertrand, Patrick; Gayda, Jean-Pierre; Rao, K. Krishna

    1982-05-01

    The temperature dependence of the electron spin-lattice relaxation time T1 of the (4Fe-4S) ferredoxin from Bacillus stearothermophilus is studied in the range 1.2 to 40 K. This dependence is similar to that observed for the (2Fe-2S) ferredoxin from Spirulina maxima and can be interpreted with the same relaxation processes [J.P. Gayda, P. Bertrand, A. Deville, C. More, G. Roger, J.F. Gibson, and R. Cammack, Biochim. Biophys. Acta 581, 15 (1979)]. In particular, between 4 and 15 K, the data are well fitted by a second-order Raman process involving three-dimensional phonons, with a Debye temperature of about 60 K (45 cm-1). This would give an estimation of the highest frequency of the vibrations which can propagate through the three-dimensional proteinic medium. In the highest temperature range (T≳30 K) the results are interpreted with an Orbach process involving an excited level of energy 120 cm-1. This process could be induced by the localized vibrations of the active site. Finally, these results are compared to those recently reported for some hemoproteins [H.J. Stapleton, J.P. Allen, C.P. Flynn, D.G. Stinson, and S.R. Kurtz, Phys. Rev. Lett. 45, 1456 (1980)]. Below 15 K, the temperature dependence of T1 for these samples is similar to that observed for the iron-sulfur proteins and may be interpreted in the same way. Our interpretation is compared to the fractal model proposed by Stapleton et al.

  18. Inhibition of IspH, a [4Fe-4S]2+ enzyme involved in the biosynthesis of isoprenoids via the methylerythritol phosphate pathway.

    PubMed

    Janthawornpong, Karnjapan; Krasutsky, Sergiy; Chaignon, Philippe; Rohmer, Michel; Poulter, C Dale; Seemann, Myriam

    2013-02-01

    The MEP pathway, which is absent in animals but present in most pathogenic bacteria, in the parasite responsible for malaria and in plant plastids, is a target for the development of antimicrobial drugs. IspH, an oxygen-sensitive [4Fe-4S] enzyme, catalyzes the last step of this pathway and converts (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate (HMBPP) into the two isoprenoid precursors: isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). A crucial step in the mechanism of this enzyme is the binding of the C4 hydroxyl of HMBPP to the unique fourth iron site in the [4Fe-4S](2+) moiety. Here, we report the synthesis and the kinetic investigations of two new extremely potent inhibitors of E. coli IspH where the OH group of HMBPP is replaced by an amino and a thiol group. (E)-4-Mercapto-3-methylbut-2-en-1-yl diphosphate is a reversible tight-binding inhibitor of IspH with K(i) = 20 ± 2 nM. A detailed kinetic analysis revealed that (E)-4-amino-3-methylbut-2-en-1-yl diphosphate is a reversible slow-binding inhibitor of IspH with K(i) = 54 ± 19 nM. The slow binding behavior of this inhibitor is best described by a one-step mechanism with the slow step consisting of the formation of the enzyme-inhibitor (EI) complex. PMID:23316732

  19. Clustering PPI data by combining FA and SHC method

    PubMed Central

    2015-01-01

    Clustering is one of main methods to identify functional modules from protein-protein interaction (PPI) data. Nevertheless traditional clustering methods may not be effective for clustering PPI data. In this paper, we proposed a novel method for clustering PPI data by combining firefly algorithm (FA) and synchronization-based hierarchical clustering (SHC) algorithm. Firstly, the PPI data are preprocessed via spectral clustering (SC) which transforms the high-dimensional similarity matrix into a low dimension matrix. Then the SHC algorithm is used to perform clustering. In SHC algorithm, hierarchical clustering is achieved by enlarging the neighborhood radius of synchronized objects continuously, while the hierarchical search is very difficult to find the optimal neighborhood radius of synchronization and the efficiency is not high. So we adopt the firefly algorithm to determine the optimal threshold of the neighborhood radius of synchronization automatically. The proposed algorithm is tested on the MIPS PPI dataset. The results show that our proposed algorithm is better than the traditional algorithms in precision, recall and f-measure value. PMID:25707632

  20. Organometallic mechanism of action and inhibition of the 4Fe-4S isoprenoid biosynthesis protein GcpE (IspG)

    PubMed Central

    Wang, Weixue; Li, Jikun; Wang, Ke; Huang, Cancan; Zhang, Yong; Oldfield, Eric

    2010-01-01

    We report the results of a series of chemical, EPR, ENDOR, and HYSCORE spectroscopic investigations of the mechanism of action (and inhibition) of GcpE, E-1-hydroxy-2-methyl-but-2-enyl-4-diphosphate (HMBPP) synthase, also known as IspG, an Fe4S4 cluster-containing protein. We find that the epoxide of HMBPP when reduced by GcpE generates the same transient EPR species as observed on addition of the substrate, 2-C-methyl-D-erythritol-2, 4-cyclo-diphosphate. ENDOR and HYSCORE spectra of these transient species (using 2H, 13C and 17O labeled samples) indicate formation of an Fe-C-H containing organometallic intermediate, most likely a ferraoxetane. This is then rapidly reduced to a ferracyclopropane in which the HMBPP product forms an η2-alkenyl π- (or π/σ) complex with the 4th Fe in the Fe4S4 cluster, and a similar “metallacycle” also forms between isopentenyl diphosphate (IPP) and GcpE. Based on this metallacycle concept, we show that an alkyne (propargyl) diphosphate is a good (Ki ∼ 300 nM) GcpE inhibitor, and supported again by EPR and ENDOR results (a 13C hyperfine coupling of ∼7 MHz), as well as literature precedent, we propose that the alkyne forms another π/σ metallacycle, an η2-alkynyl, or ferracyclopropene. Overall, the results are of broad general interest because they provide new mechanistic insights into GcpE catalysis and inhibition, with organometallic bond formation playing, in both cases, a key role. PMID:20534554

  1. Orientations of Iron-Sulfur Clusters FA and FB in the Homodimeric Type-I Photosynthetic Reaction Center of Heliobacterium modesticaldum.

    PubMed

    Kondo, Toru; Matsuoka, Masahiro; Azai, Chihiro; Itoh, Shigeru; Oh-Oka, Hirozo

    2016-05-12

    Orientations of the FA and FB iron-sulfur (FeS) clusters in a structure-unknown type-I homodimeric heriobacterial reaction center (hRC) were studied in oriented membranes of the thermophilic anaerobic photosynthetic bacterium Heliobacterium modesticaldum by electron paramagnetic resonance (EPR), and compared with those in heterodimeric photosystem I (PS I). The Rieske-type FeS center in the cytochrome b/c complex showed a well-oriented EPR signal. Illumination at 14 K induced an FB(-) signal with g-axes of gz = 2.066, gy = 1.937, and gx = 1.890, tilted at angles of 60°, 60°, and 45°, respectively, with respect to the membrane normal. Chemical reduction with dithionite produced an additional signal of FA(-), which magnetically interacted with FB(-), with gz = 2.046, gy = 1.942, and gx = 1.911 at 30°, 60°, and 90°, respectively. The angles and redox properties of FA(-) and FB(-) in hRC resemble those of FB(-) and FA(-), respectively, in PS I. Therefore, FA and FB in hRC, named after their g-value similarities, seem to be located like FB and FA, not like FA and FB, respectively, in PS I. The reducing side of hRC could resemble those in PS I, if the names of FA and FB are interchanged with each other. PMID:27101081

  2. Spectroscopic and functional characterization of iron-sulfur cluster-bound forms of Azotobacter vinelandii (Nif)IscA.

    PubMed

    Mapolelo, Daphne T; Zhang, Bo; Naik, Sunil G; Huynh, Boi Hanh; Johnson, Michael K

    2012-10-16

    The mechanism of [4Fe-4S] cluster assembly on A-type Fe-S cluster assembly proteins, in general, and the specific role of (Nif)IscA in the maturation of nitrogen fixation proteins are currently unknown. To address these questions, in vitro spectroscopic studies (UV-visible absorption/CD, resonance Raman and Mössbauer) have been used to investigate the mechanism of [4Fe-4S] cluster assembly on Azotobacter vinelandii(Nif)IscA, and the ability of (Nif)IscA to accept clusters from NifU and to donate clusters to the apo form of the nitrogenase Fe-protein. The results show that (Nif)IscA can rapidly and reversibly cycle between forms containing one [2Fe-2S](2+) and one [4Fe-4S](2+) cluster per homodimer via DTT-induced two-electron reductive coupling of two [2Fe-2S](2+) clusters and O(2)-induced [4Fe-4S](2+) oxidative cleavage. This unique type of cluster interconversion in response to cellular redox status and oxygen levels is likely to be important for the specific role of A-type proteins in the maturation of [4Fe-4S] cluster-containing proteins under aerobic growth or oxidative stress conditions. Only the [4Fe-4S](2+)-(Nif)IscA was competent for rapid activation of apo-nitrogenase Fe protein under anaerobic conditions. Apo-(Nif)IscA was shown to accept clusters from [4Fe-4S] cluster-bound NifU via rapid intact cluster transfer, indicating a potential role as a cluster carrier for delivery of clusters assembled on NifU. Overall the results support the proposal that A-type proteins can function as carrier proteins for clusters assembled on U-type proteins and suggest that they are likely to supply [2Fe-2S] clusters rather than [4Fe-4S] for the maturation of [4Fe-4S] cluster-containing proteins under aerobic or oxidative stress growth conditions. PMID:23003323

  3. Active photosynthesis in cyanobacterial mutants with directed modifications in the ligands for two iron-sulfur clusters on the PsaC protein of photosystem I.

    PubMed Central

    Mannan, R M; He, W Z; Metzger, S U; Whitmarsh, J; Malkin, R; Pakrasi, H B

    1996-01-01

    The PsaC protein of the Photosystem I (PSI) complex in thylakoid membranes coordinates two [4Fe-4S] clusters, FA and FB. Although it is known that PsaC participates in electron transfer to ferredoxin, the pathway of electrons through this protein is unknown. To elucidate the roles of FA and FB, we created two site-directed mutant strains of the cyanobacterium Anabaena variabilis ATCC 29413. In one mutant, cysteine 13, a ligand for FB was replaced by an aspartic acid (C13D); in the other mutant, cysteine 50, a ligand for FA was modified similarly (C50D). Low-temperature electron paramagnetic resonance studies demonstrated that the C50D mutant has a normal FB center and a modified FA center. In contrast, the C13D strain has normal FA, but failed to reveal any signal from FB. Room-temperature optical studies showed that C13D has only one functional electron acceptor in PsaC, whereas two such acceptors are functional in the C50D and wild-type strains. Although both mutants grow under photoautotrophic conditions, the rate of PSI-mediated electron transfer in C13D under low light levels is about half that of C50D or wild type. These data show that (i) FB is not essential for the assembly of the PsaC protein in PSI and (ii) FB is not absolutely required for electron transfer from the PSI reaction center to ferredoxin. PMID:8617228

  4. Copper binding in IscA inhibits iron-sulfur cluster assembly in Escherichia coli

    PubMed Central

    Tan, Guoqiang; Cheng, Zishuo; Pang, Yilin; Landry, Aaron P.; Li, Jianghui; Lu, Jianxin; Ding, Huangen

    2014-01-01

    Among the iron-sulfur cluster assembly proteins encoded by gene cluster iscSUA-hscBA-fdx in Escherichia coli, IscA has a unique and strong iron binding activity and can provide iron for iron-sulfur cluster assembly in proteins in vitro. Deletion of IscA and its paralogue SufA results in an E. coli mutant that fails to assemble [4Fe-4S] clusters in proteins under aerobic conditions, suggesting that IscA has a crucial role for iron-sulfur cluster biogenesis. Here we report that among the iron-sulfur cluster assembly proteins, IscA also has a strong and specific binding activity for Cu(I) in vivo and in vitro. The Cu(I) center in IscA is stable and resistant to oxidation under aerobic conditions. Mutation of the conserved cysteine residues that are essential for the iron binding in IscA abolishes the copper binding activity, indicating that copper and iron may share the same binding site in the protein. Additional studies reveal that copper can compete with iron for the metal binding site in IscA and effectively inhibits the IscA-mediated [4Fe-4S] cluster assembly in E. coli cells. The results suggest that copper may not only attack the [4Fe-4S] clusters in dehydratases, but also block the [4Fe-4S] cluster assembly in proteins by targeting IscA in cells. PMID:24946160

  5. Role of Nfu1 and Bol3 in iron-sulfur cluster transfer to mitochondrial clients.

    PubMed

    Melber, Andrew; Na, Un; Vashisht, Ajay; Weiler, Benjamin D; Lill, Roland; Wohlschlegel, James A; Winge, Dennis R

    2016-01-01

    Iron-sulfur (Fe-S) clusters are essential for many cellular processes, ranging from aerobic respiration, metabolite biosynthesis, ribosome assembly and DNA repair. Mutations in NFU1 and BOLA3 have been linked to genetic diseases with defects in mitochondrial Fe-S centers. Through genetic studies in yeast, we demonstrate that Nfu1 functions in a late step of [4Fe-4S] cluster biogenesis that is of heightened importance during oxidative metabolism. Proteomic studies revealed Nfu1 physical interacts with components of the ISA [4Fe-4S] assembly complex and client proteins that need [4Fe-4S] clusters to function. Additional studies focused on the mitochondrial BolA proteins, Bol1 and Bol3 (yeast homolog to human BOLA3), revealing that Bol1 functions earlier in Fe-S biogenesis with the monothiol glutaredoxin, Grx5, and Bol3 functions late with Nfu1. Given these observations, we propose that Nfu1, assisted by Bol3, functions to facilitate Fe-S transfer from the biosynthetic apparatus to the client proteins preventing oxidative damage to [4Fe-4S] clusters. PMID:27532773

  6. Role of Nfu1 and Bol3 in iron-sulfur cluster transfer to mitochondrial clients

    PubMed Central

    Melber, Andrew; Na, Un; Vashisht, Ajay; Weiler, Benjamin D; Lill, Roland; Wohlschlegel, James A; Winge, Dennis R

    2016-01-01

    Iron-sulfur (Fe-S) clusters are essential for many cellular processes, ranging from aerobic respiration, metabolite biosynthesis, ribosome assembly and DNA repair. Mutations in NFU1 and BOLA3 have been linked to genetic diseases with defects in mitochondrial Fe-S centers. Through genetic studies in yeast, we demonstrate that Nfu1 functions in a late step of [4Fe-4S] cluster biogenesis that is of heightened importance during oxidative metabolism. Proteomic studies revealed Nfu1 physical interacts with components of the ISA [4Fe-4S] assembly complex and client proteins that need [4Fe-4S] clusters to function. Additional studies focused on the mitochondrial BolA proteins, Bol1 and Bol3 (yeast homolog to human BOLA3), revealing that Bol1 functions earlier in Fe-S biogenesis with the monothiol glutaredoxin, Grx5, and Bol3 functions late with Nfu1. Given these observations, we propose that Nfu1, assisted by Bol3, functions to facilitate Fe-S transfer from the biosynthetic apparatus to the client proteins preventing oxidative damage to [4Fe-4S] clusters. DOI: http://dx.doi.org/10.7554/eLife.15991.001 PMID:27532773

  7. A mixed-ligand iron-sulfur cluster (C556SPaB or C565SPsaB) in the Fx-binding site leads to a decreased quantum efficiency of electron transfer in photosystem I.

    PubMed Central

    Vassiliev, I R; Jung, Y S; Smart, L B; Schulz, R; McIntosh, L; Golbeck, J H

    1995-01-01

    The proposed structure of Photosystem I depicts two cysteines on the PsaA polypeptide and two cysteines on the PsaB polypeptide in a symmetrical environment, each providing ligands for the interpolypeptide Fx cluster. We studied the role of Fx in electron transfer by substituting serine for cysteine (C565SPsaB and C556SPsaB), thereby introducing the first example of a genetically engineered, mixed-ligand [4Fe-4S] cluster into a protein. Optical kinetic spectroscopy shows that after a single-turnover flash at 298 K, the contribution of A1- (lifetime of 10 microseconds, 40% of total and lifetime of 100 microseconds, 20% of total) and Fx- (lifetime of 500-800 microseconds, 10-15% of total) to the overall P700+ back reaction have increased in C565SPsaB and C556SPsaB at the expense of the back reaction from [FA/FB]-. The electron paramagnetic resonance spectrum of Fx shows g-values of 2.04, 1.94, and 1.81 in both mutants and a similarly decreased amount of FA and FB reduced at 15 K after a single-turnover flash. These results indicate that the mixed-ligand (3 cysteines, 1 serine) Fx cluster is an inefficient electron carrier, but that a small leak through Fx still permits FA and FB to be reduced quantitatively when the samples are frozen during continuous illumination. The data confirm that Fx is a necessary intermediate in the electron transfer pathway from A1 to FA and FB in Photosystem I. PMID:8534825

  8. New redox states observed in [FeFe] hydrogenases reveal redox coupling within the H-cluster.

    PubMed

    Adamska-Venkatesh, Agnieszka; Krawietz, Danuta; Siebel, Judith; Weber, Katharina; Happe, Thomas; Reijerse, Edward; Lubitz, Wolfgang

    2014-08-13

    Active [FeFe] hydrogenases can be obtained by expressing the unmaturated enzyme in Escherichia coli followed by incubation with a synthetic precursor of the binuclear [2Fe] subcluster, namely: [NEt4]2[Fe2(adt)(CO)4(CN)2] (adt = [S-CH2-NH-CH2-S](2-)). The binuclear subsite Fe2(adt)(CO)3(CN)2 is attached through a bridging cysteine side chain to a [4Fe-4S] subcluster already present in the unmaturated enzyme thus yielding the intact native "H-cluster". We present FTIR electrochemical studies of the [FeFe] hydrogenase from Chlamydomonas reinhardtii, CrHydA1, maturated with the precursor of the native cofactor [Fe2(adt)(CO)4(CN)2](2-) as well as a non-natural variant [Fe2(pdt)(CO)4(CN)2](2-) in which the bridging amine functionality is replaced by CH2. The obtained active enzyme CrHydA1(adt) shows the same redox states in the respective potential range as observed for the native system (E(ox/red) = -400 mV, E(red/sred) = -470 mV). For the Hox → Hred transition the reducing equivalent is stored on the binuclear part, ([4Fe-4S](2+)Fe(II)Fe(I) → [4Fe-4S](2+)Fe(I)Fe(I)), while the Hred → Hsred transition is characterized by a reduction of the [4Fe-4S] part of the H-cluster ([4Fe-4S](2+)Fe(I)Fe(I) → [4Fe-4S](+)Fe(I)Fe(I)). A similar transition is reported here for the CO inhibited state of the H-cluster: ([4Fe-4S](2+)Fe(I)Fe(II)CO → [4Fe-4S](+)Fe(I)Fe(II)CO). An FTIR electrochemical study of the inactive variant with the pdt ligand, CrHydA1(pdt), identified two redox states H(pdt)-ox and H(pdt)-"red". Both EPR and FTIR spectra of H(pdt)-ox are virtually identical to those of the H(adt)-ox and the native Hox state. The H(pdt)-"red" state is also characterized by a reduced [4Fe-4S] subcluster. In contrast to CrHydA1(adt), the H(pdt)-ox state of CrHydA1(pdt) is stable up to rather high potentials (+200 mV). This study demonstrates the distinct redox coupling between the two parts of the H-cluster and confirms that the [4Fe-4S]H subsite is also redox active and as

  9. FA-SIFT study of reactions of protonated water and ethanol clusters with [alpha]-pinene and linalool in view of their selective detection by CIMS

    NASA Astrophysics Data System (ADS)

    Dhooghe, F.; Amelynck, C.; Rimetz-Planchon, J.; Schoon, N.; Vanhaecke, F.

    2010-02-01

    The use of protonated water clusters and protonated ethanol clusters as reagent ions has been evaluated for the resolution of an interference encountered in CIMS when measuring monoterpenes (C10H16) and linalool (C10H18O) simultaneously. To this end, the reactions of H3O+.(H2O)n (n = 1-3), (C2H5OH)mH+ (m = 1-3) and (C2H5OH.H2O)H+ with [alpha]-pinene and linalool have been characterized in a flowing afterglow-selected ion flow tube (FA-SIFT) instrument at a SIFT He buffer gas pressure of 1.43 hPa and a temperature of 298 K. All reactions with linalool were found to occur at the collision limit. The reaction of (C2H5OH)2H+ with [alpha]-pinene proceeds at half the collision rate and both the reactions of (C2H5OH)3H+ and H3O+.(H2O)3 with [alpha]-pinene have a very low rate constant. All other reactions involving [alpha]-pinene proceed at the collision rate. The reactions of H3O+.H2O, H3O+.(H2O)2, C2H5OH2+, (C2H5OH.H2O)H+ and (C2H5OH)2H+ with [alpha]-pinene mainly proceed by proton transfer. Additionally, ligand switching channels have been observed for the reactions of (C2H5OH)2H+ and H3O+.(H2O)2 with [alpha]-pinene. Protonated linalool was observed as a minor product for the reactions of (C2H5OH.H2O)H+ and H3O+.(H2O)n (n = 1-3) with linalool. For all linalool reactions, a contribution of the dissociative proton transfer product at m/z 137 was found and this ion was the main product ion for the reactions with H3O+.H2O, C2H5OH2+ and (C2H5OH.H2O)H+. For the (C2H5OH.H2O)H+/linalool reaction, ligand switching with both water and ethanol has been observed. Major ligand switching channels were observed for the reactions of (C2H5OH)2H+, (C2H5OH)3H+ and H3O+.(H2O)2 with linalool. Also, for the H3O+.(H2O)3/linalool reaction, several ligand switching channels have been observed. These results are discussed in view of their applicability for the selective detection of monoterpenes and linalool with CIMS instrumentation such as SIFT-MS, PTR-MS and APCI-MS.

  10. Identification of FeS clusters in the glycyl-radical enzyme benzylsuccinate synthase via EPR and Mössbauer spectroscopy.

    PubMed

    Hilberg, Markus; Pierik, Antonio J; Bill, Eckhard; Friedrich, Thorsten; Lippert, Marie-Luise; Heider, Johann

    2012-01-01

    The anaerobic degradation pathway of toluene is initiated by the addition of the methyl group of toluene to the double bond of fumarate. This reaction is catalyzed by a novel glycyl-radical enzyme, (R)-benzylsuccinate synthase (BSS). The enzyme consists of three subunits, α, β, and γ, and differs from most other glycyl-radical enzymes in having additional cofactors. We have purified a Strep-tagged nonactivated BSS from recombinant Escherichia coli and identified the additional cofactors as FeS clusters by UV/vis, EPR, and Mössbauer spectroscopy. Analysis of the metal content as well as the EPR and Mössbauer spectra indicated that BSS contains magnetically coupled low-potential [4Fe-4S] clusters. Several enzyme preparations showed differing amounts of [3Fe-4S] clusters that could be reconstituted to [4Fe-4S] clusters, indicating that they arise from partial decay of the initial [4Fe-4S] clusters. The most likely location of these FeS clusters in the enzyme are the small β and γ subunits, which are unique for the BSS subfamily of glycyl-radical enzymes and contain conserved cysteines as potential ligands. PMID:21837540

  11. Crystal Structures of the Iron-Sulfur Cluster-Dependent Quinolinate Synthase in Complex with Dihydroxyacetone Phosphate, Iminoaspartate Analogues, and Quinolinate.

    PubMed

    Fenwick, Michael K; Ealick, Steven E

    2016-08-01

    The quinolinate synthase of prokaryotes and photosynthetic eukaryotes, NadA, contains a [4Fe-4S] cluster with unknown function. We report crystal structures of Pyrococcus horikoshii NadA in complex with dihydroxyacetone phosphate (DHAP), iminoaspartate analogues, and quinolinate. DHAP adopts a nearly planar conformation and chelates the [4Fe-4S] cluster via its keto and hydroxyl groups. The active site architecture suggests that the cluster acts as a Lewis acid in enediolate formation, like zinc in class II aldolases. The DHAP and putative iminoaspartate structures suggest a model for a condensed intermediate. The ensemble of structures suggests a two-state system, which may be exploited in early steps. PMID:27404889

  12. The FX iron-sulfur cluster serves as the terminal bound electron acceptor in heliobacterial reaction centers.

    PubMed

    Romberger, Steven P; Golbeck, John H

    2012-03-01

    Phototrophs of the family Heliobacteriaceae contain the simplest known Type I reaction center (RC), consisting of a homodimeric (PshA)(2) core devoid of bound cytochromes and antenna proteins. Unlike plant and cyanobacterial Photosystem I in which the F(A)/F(B) protein, PsaC, is tightly bound to P(700)-F(X) cores, the RCs of Heliobacterium modesticaldum contain two F(A)/F(B) proteins, PshBI and PshBII, which are loosely bound to P(800)-F(X) cores. These two 2[4Fe-4S] ferredoxins have been proposed to function as mobile redox proteins, reducing downstream metabolic partners much in the same manner as does [2Fe-2S] ferredoxin or flavodoxin (Fld) in PS I. Using P(800)-F(X) cores devoid of PshBI and PshBII, we show that iron-sulfur cluster F(X) directly reduces Fld without the involvement of F(A) or F(B) (Fld is used as a proxy for soluble redox proteins even though a gene encoding Fld is not identified in the H. modesticaldum genome). The reduction of Fld is suppressed by the addition of PshBI or PshBII, an effect explained by competition for the electron on F(X). In contrast, P(700)-F(X) cores require the presence of the PsaC, and hence, the F(A)/F(B) clusters for Fld (or ferredoxin) reduction. Thus, in H. modesticaldum, the interpolypeptide F(X) cluster serves as the terminal bound electron acceptor. This finding implies that the homodimeric (PshA)(2) cores should be capable of donating electrons to a wide variety of yet-to-be characterized soluble redox partners. PMID:22297911

  13. Crystal structure of an Fe-S cluster-containing fumarate hydratase enzyme from Leishmania major reveals a unique protein fold.

    PubMed

    Feliciano, Patricia R; Drennan, Catherine L; Nonato, M Cristina

    2016-08-30

    Fumarate hydratases (FHs) are essential metabolic enzymes grouped into two classes. Here, we present the crystal structure of a class I FH, the cytosolic FH from Leishmania major, which reveals a previously undiscovered protein fold that coordinates a catalytically essential [4Fe-4S] cluster. Our 2.05 Å resolution data further reveal a dimeric architecture for this FH that resembles a heart, with each lobe comprised of two domains that are arranged around the active site. Besides the active site, where the substrate S-malate is bound bidentate to the unique iron of the [4Fe-4S] cluster, other binding pockets are found near the dimeric enzyme interface, some of which are occupied by malonate, shown here to be a weak inhibitor of this enzyme. Taken together, these data provide a framework both for investigations of the class I FH catalytic mechanism and for drug design aimed at fighting neglected tropical diseases. PMID:27528683

  14. Characterization of a unique [FeS] cluster in the electron transfer chain of the oxygen tolerant [NiFe] hydrogenase from Aquifex aeolicus

    PubMed Central

    Pandelia, Maria-Eirini; Nitschke, Wolfgang; Infossi, Pascale; Giudici-Orticoni, Marie-Thérèse; Bill, Eckhard; Lubitz, Wolfgang

    2011-01-01

    Iron-sulfur clusters are versatile electron transfer cofactors, ubiquitous in metalloenzymes such as hydrogenases. In the oxygen-tolerant Hydrogenase I from Aquifex aeolicus such electron “wires” form a relay to a diheme cytb, an integral part of a respiration pathway for the reduction of O2 to water. Amino acid sequence comparison with oxygen-sensitive hydrogenases showed conserved binding motifs for three iron-sulfur clusters, the nature and properties of which were unknown so far. Electron paramagnetic resonance spectra exhibited complex signals that disclose interesting features and spin-coupling patterns; by redox titrations three iron-sulfur clusters were identified in their usual redox states, a [3Fe4S] and two [4Fe4S], but also a unique high-potential (HP) state was found. On the basis of 57Fe Mössbauer spectroscopy we attribute this HP form to a superoxidized state of the [4Fe4S] center proximal to the [NiFe] site. The unique environment of this cluster, characterized by a surplus cysteine coordination, is able to tune the redox potentials and make it compliant with the [4Fe4S]3+ state. It is actually the first example of a biological [4Fe4S] center that physiologically switches between 3+, 2+, and 1+ oxidation states within a very small potential range. We suggest that the (1 + /2+) redox couple serves the classical electron transfer reaction, whereas the superoxidation step is associated with a redox switch against oxidative stress. PMID:21444783

  15. Frataxin Accelerates [2Fe-2S] Cluster Formation on the Human Fe–S Assembly Complex

    PubMed Central

    Fox, Nicholas G.; Das, Deepika; Chakrabarti, Mrinmoy; Lindahl, Paul A.; Barondeau, David P.

    2015-01-01

    Iron–sulfur (Fe–S) clusters function as protein cofactors for a wide variety of critical cellular reactions. In human mitochondria, a core Fe–S assembly complex [called SDUF and composed of NFS1, ISD11, ISCU2, and frataxin (FXN) proteins] synthesizes Fe–S clusters from iron, cysteine sulfur, and reducing equivalents and then transfers these intact clusters to target proteins. In vitro assays have relied on reducing the complexity of this complicated Fe–S assembly process by using surrogate electron donor molecules and monitoring simplified reactions. Recent studies have concluded that FXN promotes the synthesis of [4Fe-4S] clusters on the mammalian Fe–S assembly complex. Here the kinetics of Fe–S synthesis reactions were determined using different electron donation systems and by monitoring the products with circular dichroism and absorbance spectroscopies. We discovered that common surrogate electron donor molecules intercepted Fe–S cluster intermediates and formed high-molecular weight species (HMWS). The HMWS are associated with iron, sulfide, and thiol-containing proteins and have properties of a heterogeneous solubilized mineral with spectroscopic properties remarkably reminiscent of those of [4Fe-4S] clusters. In contrast, reactions using physiological reagents revealed that FXN accelerates the formation of [2Fe-2S] clusters rather than [4Fe-4S] clusters as previously reported. In the preceding paper [Fox, N. G., et al. (2015) Biochemistry 54, DOI: 10.1021/bi5014485], [2Fe-2S] intermediates on the SDUF complex were shown to readily transfer to uncomplexed ISCU2 or apo acceptor proteins, depending on the reaction conditions. Our results indicate that FXN accelerates a rate-limiting sulfur transfer step in the synthesis of [2Fe-2S] clusters on the human Fe–S assembly complex. PMID:26016518

  16. Spectroscopic changes during a single turnover of biotin synthase: destruction of a [2Fe-2S] cluster accompanies sulfur insertion.

    PubMed

    Ugulava, N B; Sacanell, C J; Jarrett, J T

    2001-07-27

    Biotin synthase catalyzes the insertion of a sulfur atom between the saturated C6 and C9 carbons of dethiobiotin. Catalysis requires AdoMet and flavodoxin and generates 5'-deoxyadenosine and methionine, suggesting that biotin synthase is an AdoMet-dependent radical enzyme. Biotin synthase (BioB) is aerobically purified as a dimer of 38.4 kDa monomers that contains 1-1.5 [2Fe-2S](2+) clusters per monomer and can be reconstituted with exogenous iron, sulfide, and reductants to contain up to two [4Fe-4S] clusters per monomer. The iron-sulfur clusters may play a dual role in biotin synthase: a reduced iron-sulfur cluster is probably involved in radical generation by mediating the reductive cleavage of AdoMet, while recent in vitro labeling studies suggest that an iron-sulfur cluster also serves as the immediate source of sulfur for the biotin thioether ring. Consistent with this dual role for iron-sulfur clusters in biotin synthase, we have found that the protein is stable, containing one [2Fe-2S](2+) cluster and one [4Fe-4S](2+) cluster per monomer. In the present study, we demonstrate that this mixed cluster state is essential for optimal activity. We follow changes in the Fe and S content and UV/visible and EPR spectra of the enzyme during a single turnover and conclude that during catalysis the [4Fe-4S](2+) cluster is preserved while the [2Fe-2S](2+) cluster is destroyed. We propose a mechanism for incorporation of sulfur into dethiobiotin in which a sulfur atom is oxidatively extracted from the [2Fe-2S](2+) cluster. PMID:11444982

  17. The Human Iron–Sulfur Assembly Complex Catalyzes the Synthesis of [2Fe-2S] Clusters on ISCU2 That Can Be Transferred to Acceptor Molecules

    PubMed Central

    Fox, Nicholas G.; Chakrabarti, Mrinmoy; McCormick, Sean P.; Lindahl, Paul A.; Barondeau, David P.

    2015-01-01

    Iron–sulfur (Fe–S) clusters are essential protein cofactors for most life forms. In human mitochondria, the core Fe–S biosynthetic enzymatic complex (called SDUF) consists of NFS1, ISD11, ISCU2, and frataxin (FXN) protein components. Few mechanistic details about how this complex synthesizes Fe–S clusters and how these clusters are delivered to targets are known. Here circular dichroism and Mössbauer spectroscopies were used to reveal details of the Fe–S cluster assembly reaction on the SDUF complex. SDUF reactions generated [2Fe-2S] cluster intermediates that readily converted to stable [2Fe-2S] clusters bound to uncomplexed ISCU2. Similar reactions that included the apo Fe–S acceptor protein human ferredoxin (FDX1) resulted in formation of [2Fe-2S]-ISCU2 rather than [2Fe-2S]-FDX1. Subsequent addition of dithiothreitol (DTT) induced transfer of the cluster from ISCU2 to FDX1, suggesting that [2Fe-2S]-ISCU2 is an intermediate. Reactions that initially included DTT rapidly generated [2Fe-2S]-FDX1 and bypassed formation of [2Fe-2S]-ISCU2. In the absence of apo-FDX1, incubation of [2Fe-2S]-ISCU2 with DTT generated [4Fe-4S]-ISCU2 species. Together, these results conflict with a recent report of stable [4Fe-4S] cluster formation on the SDUF complex. Rather, they support a model in which SDUF builds transient [2Fe-2S] cluster intermediates that generate clusters on sulfur-containing molecules, including uncomplexed ISCU2. Additional small molecule or protein factors are required for the transfer of these clusters to Fe–S acceptor proteins or the synthesis of [4Fe-4S] clusters. PMID:26016389

  18. X-ray crystallographic and EPR spectroscopic analysis of HydG, a maturase in [FeFe]-hydrogenase H-cluster assembly

    PubMed Central

    Dinis, Pedro; Suess, Daniel L. M.; Fox, Stephen J.; Harmer, Jenny E.; Driesener, Rebecca C.; De La Paz, Liliana; Swartz, James R.; Essex, Jonathan W.; Britt, R. David; Roach, Peter L.

    2015-01-01

    Hydrogenases use complex metal cofactors to catalyze the reversible formation of hydrogen. In [FeFe]-hydrogenases, the H-cluster cofactor includes a diiron subcluster containing azadithiolate, three CO, and two CN− ligands. During the assembly of the H cluster, the radical S-adenosyl methionine (SAM) enzyme HydG lyses the substrate tyrosine to yield the diatomic ligands. These diatomic products form an enzyme-bound Fe(CO)x(CN)y synthon that serves as a precursor for eventual H-cluster assembly. To further elucidate the mechanism of this complex reaction, we report the crystal structure and EPR analysis of HydG. At one end of the HydG (βα)8 triosephosphate isomerase (TIM) barrel, a canonical [4Fe-4S] cluster binds SAM in close proximity to the proposed tyrosine binding site. At the opposite end of the active-site cavity, the structure reveals the auxiliary Fe-S cluster in two states: one monomer contains a [4Fe-5S] cluster, and the other monomer contains a [5Fe-5S] cluster consisting of a [4Fe-4S] cubane bridged by a μ2-sulfide ion to a mononuclear Fe2+ center. This fifth iron is held in place by a single highly conserved protein-derived ligand: histidine 265. EPR analysis confirms the presence of the [5Fe-5S] cluster, which on incubation with cyanide, undergoes loss of the labile iron to yield a [4Fe-4S] cluster. We hypothesize that the labile iron of the [5Fe-5S] cluster is the site of Fe(CO)x(CN)y synthon formation and that the limited bonding between this iron and HydG may facilitate transfer of the intact synthon to its cognate acceptor for subsequent H-cluster assembly. PMID:25605932

  19. Exchange variation of zero-field splittings in [Fe 4S 4] + clusters of ferredoxins with high-spin S=3/2 ground state

    NASA Astrophysics Data System (ADS)

    Belinsky, Moisey I.

    2000-04-01

    The spin-coupling model of zero-field splitting (ZFS) of tetrameric mixed-valent and monovalent clusters is developed. The <| ŝi0(2)|> matrix elements of the individual ZFS tensor operator of the second rank were calculated for tetramers of the general type in the representation of the total and intermediate spins. The spin-dependent correlations between ZFS parameters D S of the cluster states S and D i of individual ions s i were obtained for [4Fe-4S] + centers of native systems and synthetic model compounds with the high-spin Sgr=3/2 ground state. It was shown that the single-particle spin parameters f i=<| ŝiz2-s i(s i+1)/3|>/<| Ŝz2-S(S+1)/3|> of the [Fe 3(II)Fe(III)] center essentially depend on the total, intermediate and local spins and on the exchange parameters. In the spin schemes with fixed intermediate spins, the correlation between the cluster total spin ZFS parameters, D S, and the individual ZFS parameters, D i, are determined by the total, intermediate and individual spins. Anisotropic cluster ZFS parameters D S strongly depend on isotropic Heisenberg exchange and double exchange inter-ion interactions due to exchange mixing of the states with different intermediate spins. It was shown that the cluster ZFS parameters D S, for the states with S=3/2, change their values and sign under the variation of the Heisenberg exchange and double exchange parameters during the cluster deformation. The single-particle ZFS parameters D i and the exchange effects determine the observed strong positive ( DS=1.5-6 cm -1) and negative ( DS=-1.7 to -5 cm -1) cluster ZFS splittings of the ground Sgr=3/2 states of the [4Fe-4S] + clusters. Positive (negative) cluster ZFS parameters D S correspond to negative (positive) individual ZFS parameters D1-3 [Fe(II)]. The theory explains the observation of small negative effective hyperfine constants A i for the Sgr=3/2 clusters with positive and negative cluster ZFS parameters D S. The correlations between the individual and

  20. Bacterial iron-sulfur cluster sensors in mammalian pathogens

    PubMed Central

    Miller, Halie K.; Auerbuch, Victoria

    2015-01-01

    Iron-sulfur clusters act as important cofactors for a number of transcriptional regulators in bacteria, including many mammalian pathogens. The sensitivity of iron-sulfur clusters to iron availability, oxygen tension, and reactive oxygen and nitrogen species enables bacteria to use such regulators to adapt their gene expression profiles rapidly in response to changing environmental conditions. In this review, we discuss how the [4Fe-4S] or [2Fe-2S] cluster-containing regulators FNR, Wbl, aconitase, IscR, NsrR, SoxR, and AirSR contribute to bacterial pathogenesis through control of both metabolism and classical virulence factors. In addition, we briefly review mammalian iron homeostasis as well as oxidative/nitrosative stress to provide context for understanding the function of bacterial iron-sulfur cluster sensors in different niches within the host. PMID:25738802

  1. Iron-sulfur clusters as biological sensors: the chemistry of reactions with molecular oxygen and nitric oxide.

    PubMed

    Crack, Jason C; Green, Jeffrey; Thomson, Andrew J; Le Brun, Nick E

    2014-10-21

    Iron-sulfur cluster proteins exhibit a range of physicochemical properties that underpin their functional diversity in biology, which includes roles in electron transfer, catalysis, and gene regulation. Transcriptional regulators that utilize iron-sulfur clusters are a growing group that exploit the redox and coordination properties of the clusters to act as sensors of environmental conditions including O2, oxidative and nitrosative stress, and metabolic nutritional status. To understand the mechanism by which a cluster detects such analytes and then generates modulation of DNA-binding affinity, we have undertaken a combined strategy of in vivo and in vitro studies of a range of regulators. In vitro studies of iron-sulfur cluster proteins are particularly challenging because of the inherent reactivity and fragility of the cluster, often necessitating strict anaerobic conditions for all manipulations. Nevertheless, and as discussed in this Account, significant progress has been made over the past decade in studies of O2-sensing by the fumarate and nitrate reduction (FNR) regulator and, more recently, nitric oxide (NO)-sensing by WhiB-like (Wbl) and FNR proteins. Escherichia coli FNR binds a [4Fe-4S] cluster under anaerobic conditions leading to a DNA-binding dimeric form. Exposure to O2 converts the cluster to a [2Fe-2S] form, leading to protein monomerization and hence loss of DNA binding ability. Spectroscopic and kinetic studies have shown that the conversion proceeds via at least two steps and involves a [3Fe-4S](1+) intermediate. The second step involves the release of two bridging sulfide ions from the cluster that, unusually, are not released into solution but rather undergo oxidation to sulfane (S(0)) subsequently forming cysteine persulfides that then coordinate the [2Fe-2S] cluster. Studies of other [4Fe-4S] cluster proteins that undergo oxidative cluster conversion indicate that persulfide formation and coordination may be more common than previously

  2. Silver(I), Mercury(II), Cadmium(II), and Zinc(II) Target Exposed Enzymic Iron-Sulfur Clusters when They Toxify Escherichia coli

    PubMed Central

    Xu, Fang Fang

    2012-01-01

    The toxicity of soft metals is of broad interest to microbiologists, both because such metals influence the community structures in natural environments and because several metals are used as antimicrobial agents. Their potency roughly parallels their thiophilicity, suggesting that their primary biological targets are likely to be enzymes that contain key sulfhydryl moieties. A recent study determined that copper poisons Escherichia coli in part by attacking the exposed [4Fe-4S] clusters of dehydratases. The present investigation sought to test whether other soft metals also target these enzymes. In vitro experiments revealed that low-micromolar concentrations of Ag(I) and Hg(II) directly inactivated purified fumarase A, a member of the dehydratase family. The enzyme was also poisoned by higher levels of Cd(II) and Zn(II), but it was unaffected by even millimolar concentrations of Mn(II), Co(II), Ni(II), and Pb(II). Electron paramagnetic resonance analysis and measurements of released iron confirmed that damage was associated with destruction of the [4Fe-4S] cluster, and indeed, the reconstruction of the cluster fully restored activity. Growth studies were then performed to test whether dehydratase damage might underlie toxicity in vivo. Barely toxic doses of Ag(I), Hg(II), Cd(II), and Zn(II) inactivated all tested members of the [4Fe-4S] dehydratase family. Again, activity was recovered when the clusters were rebuilt. The metals did not diminish the activities of other sampled enzymes, including NADH dehydrogenase I, an iron-sulfur protein whose clusters are shielded by polypeptide. Thus, the data indicate that dehydratases are damaged by the concentrations of metals that initiate bacteriostasis. PMID:22344668

  3. O2 Reactions at the Six-iron Active Site (H-cluster) in [FeFe]-Hydrogenase*

    PubMed Central

    Lambertz, Camilla; Leidel, Nils; Havelius, Kajsa G. V.; Noth, Jens; Chernev, Petko; Winkler, Martin; Happe, Thomas; Haumann, Michael

    2011-01-01

    Irreversible inhibition by molecular oxygen (O2) complicates the use of [FeFe]-hydrogenases (HydA) for biotechnological hydrogen (H2) production. Modification by O2 of the active site six-iron complex denoted as the H-cluster ([4Fe4S]-2FeH) of HydA1 from the green alga Chlamydomonas reinhardtii was characterized by x-ray absorption spectroscopy at the iron K-edge. In a time-resolved approach, HydA1 protein samples were prepared after increasing O2 exposure periods at 0 °C. A kinetic analysis of changes in their x-ray absorption near edge structure and extended X-ray absorption fine structure spectra revealed three phases of O2 reactions. The first phase (τ1 ≤ 4 s) is characterized by the formation of an increased number of Fe–O,C bonds, elongation of the Fe–Fe distance in the binuclear unit (2FeH), and oxidation of one iron ion. The second phase (τ2 ≈ 15 s) causes a ∼50% decrease of the number of ∼2.7-Å Fe–Fe distances in the [4Fe4S] subcluster and the oxidation of one more iron ion. The final phase (τ3 ≤ 1000 s) leads to the disappearance of most Fe–Fe and Fe–S interactions and further iron oxidation. These results favor a reaction sequence, which involves 1) oxygenation at 2FeH+ leading to the formation of a reactive oxygen species-like superoxide (O2−), followed by 2) H-cluster inactivation and destabilization due to ROS attack on the [4Fe4S] cluster to convert it into an apparent [3Fe4S]+ unit, leading to 3) complete O2-induced degradation of the remainders of the H-cluster. This mechanism suggests that blocking of ROS diffusion paths and/or altering the redox potential of the [4Fe4S] cubane by genetic engineering may yield improved O2 tolerance in [FeFe]-hydrogenase. PMID:21930709

  4. Slow formation of [3Fe-4S](1+) clusters in mutant forms of Desulfovibrio africanus ferredoxin III.

    PubMed

    Hannan, J P; Busch, J L; James, R; Thomson, A J; Moore, G R; Davy, S L

    2000-02-25

    Desulfovibrio africanus ferredoxin III (Da FdIII) readily interconverts between a 7Fe and an 8Fe form with Asp-14 believed to provide a cluster ligand in the latter form. To investigate the factors important for cluster interconversion in Fe/S cluster-containing proteins we have studied two variants of Da FdIII produced by site-directed mutagenesis, Asp14Glu and Asp14His, with cluster incorporation performed in vitro. Characterisation of these proteins by UV/visible, EPR and (1)H NMR spectroscopies revealed that the formation of the stable 7Fe form of these proteins takes some time to occur. Evidence is presented which indicates the [4Fe-4S](2+) cluster is incorporated prior to the [3Fe-4S](1+) cluster. PMID:10692579

  5. A Redox Active [2Fe-2S] Cluster on the Hydrogenase Maturase HydF.

    PubMed

    Shepard, Eric M; Byer, Amanda S; Betz, Jeremiah N; Peters, John W; Broderick, Joan B

    2016-06-28

    [FeFe]-hydrogenases are nature's most prolific hydrogen catalysts, excelling at facilely interconverting H2 and protons. The catalytic core common to all [FeFe]-hydrogenases is a complex metallocofactor, referred to as the H-cluster, which is composed of a standard [4Fe-4S] cluster linked through a bridging thiolate to a 2Fe subcluster harboring dithiomethylamine, carbon monoxide, and cyanide ligands. This 2Fe subcluster is synthesized and inserted into [FeFe]-hydrogenase by three maturase enzymes denoted HydE, HydF, and HydG. HydE and HydG are radical S-adenosylmethionine enzymes and synthesize the nonprotein ligands of the H-cluster. HydF is a GTPase that functions as a scaffold or carrier for 2Fe subcluster production. Herein, we utilize UV-visible, circular dichroism, and electron paramagnetic resonance spectroscopic studies to establish the existence of redox active [4Fe-4S] and [2Fe-2S] clusters bound to HydF. We have used spectroelectrochemical titrations to assign iron-sulfur cluster midpoint potentials, have shown that HydF purifies with a reduced [2Fe-2S] cluster in the absence of exogenous reducing agents, and have tracked iron-sulfur cluster spectroscopic changes with quaternary structural perturbations. Our results provide an important foundation for understanding the maturation process by defining the iron-sulfur cluster content of HydF prior to its interaction with HydE and HydG. We speculate that the [2Fe-2S] cluster of HydF either acts as a placeholder for HydG-derived Fe(CO)2CN species or serves as a scaffold for 2Fe subcluster assembly. PMID:27232385

  6. Fanconi anemia (FA) and crosslinker sensitivity: Re-appraising the origins of FA definition.

    PubMed

    Pagano, Giovanni; d'Ischia, Marco; Pallardó, Federico V

    2015-07-01

    The commonly accepted definition of Fanconi anemia (FA) relying on DNA repair deficiency is submitted to a critical review starting from the early reports pointing to mitomycin C bioactivation and to the toxicity mechanisms of diepoxybutane and a group of nitrogen mustards causing DNA crosslinks in FA cells. A critical analysis of the literature prompts revisiting the FA phenotype and crosslinker sensitivity in terms of an oxidative stress (OS) background, redox-related anomalies of FA (FANC) proteins, and mitochondrial dysfunction. This re-appraisal of FA basic defect might lead to innovative approaches both in elucidating FA phenotypes and in clinical management. PMID:25732180

  7. GPM Video of In-fa

    NASA Video Gallery

    On Nov. 19 GPM saw a few towering storms in In-fa's eye wall were reaching heights of up to 17.3 km (10.7 miles). The most intense precipitation was measured in In-fa's eye wall by DPR where it was...

  8. Mycobacterium tuberculosis WhiB1 is an essential DNA-binding protein with a nitric oxide-sensitive iron-sulfur cluster.

    PubMed

    Smith, Laura J; Stapleton, Melanie R; Fullstone, Gavin J M; Crack, Jason C; Thomson, Andrew J; Le Brun, Nick E; Hunt, Debbie M; Harvey, Evelyn; Adinolfi, Salvatore; Buxton, Roger S; Green, Jeffrey

    2010-12-15

    Mycobacterium tuberculosis is a major pathogen that has the ability to establish, and emerge from, a persistent state. Wbl family proteins are associated with developmental processes in actinomycetes, and M. tuberculosis has seven such proteins. In the present study it is shown that the M. tuberculosis H37Rv whiB1 gene is essential. The WhiB1 protein possesses a [4Fe-4S]2+ cluster that is stable in air but reacts rapidly with eight equivalents of nitric oxide to yield two dinuclear dinitrosyl-iron thiol complexes. The [4Fe-4S] form of WhiB1 did not bind whiB1 promoter DNA, but the reduced and oxidized apo-WhiB1, and nitric oxide-treated holo-WhiB1 did bind to DNA. Mycobacterium smegmatis RNA polymerase induced transcription of whiB1 in vitro; however, in the presence of apo-WhiB1, transcription was severely inhibited, irrespective of the presence or absence of the CRP (cAMP receptor protein) Rv3676, which is known to activate whiB1 expression. Footprinting suggested that autorepression of whiB1 is achieved by apo-WhiB1 binding at a region that overlaps the core promoter elements. A model incorporating regulation of whiB1 expression in response to nitric oxide and cAMP is discussed with implications for sensing two important signals in establishing M. tuberculosis infections. PMID:20929442

  9. Nuclear resonance vibrational spectroscopy reveals the FeS cluster composition and active site vibrational properties of an O2-tolerant NAD+-reducing [NiFe] hydrogenase

    DOE PAGESBeta

    Lauterbach, Lars; Wang, Hongxin; Horch, Marius; Gee, Leland B.; Yoda, Yoshitaka; Tanaka, Yoshihito; Zebger, Ingo; Lenz, Oliver; Cramer, Stephen P.

    2014-10-30

    Hydrogenases are complex metalloenzymes that catalyze the reversible splitting of molecular hydrogen into protons and electrons essentially without overpotential. The NAD+-reducing soluble hydrogenase (SH) from Ralstonia eutropha is capable of H2 conversion even in the presence of usually toxic dioxygen. The molecular details of the underlying reactions are largely unknown, mainly because of limited knowledge of the structure and function of the various metal cofactors present in the enzyme. Here, all iron-containing cofactors of the SH were investigated by 57Fe specific nuclear resonance vibrational spectroscopy (NRVS). Our data provide experimental evidence for one [2Fe2S] center and four [4Fe4S] clusters, whichmore » is consistent with the amino acid sequence composition. Only the [2Fe2S] cluster and one of the four [4Fe4S] clusters were reduced upon incubation of the SH with NADH. This finding explains the discrepancy between the large number of FeS clusters and the small amount of FeS cluster-related signals as detected by electron paramagnetic resonance spectroscopic analysis of several NAD+-reducing hydrogenases. For the first time, Fe–CO and Fe–CN modes derived from the [NiFe] active site could be distinguished by NRVS through selective 13C labeling of the CO ligand. This strategy also revealed the molecular coordinates that dominate the individual Fe–CO modes. The present approach explores the complex vibrational signature of the Fe–S clusters and the hydrogenase active site, thereby showing that NRVS represents a powerful tool for the elucidation of complex biocatalysts containing multiple cofactors.« less

  10. IscA, an alternate scaffold for Fe-S cluster biosynthesis.

    PubMed

    Krebs, C; Agar, J N; Smith, A D; Frazzon, J; Dean, D R; Huynh, B H; Johnson, M K

    2001-11-20

    An IscA homologue within the nif regulon of Azotobacter vinelandii, designated (Nif)IscA, was expressed in Escherichia coli and purified to homogeneity. Purified (Nif)IscA was found to be a homodimer of 11-kDa subunits that contained no metal centers or other prosthetic groups in its as-isolated form. Possible roles for (Nif)IscA in Fe-S cluster biosynthesis were assessed by investigating the ability to bind iron and to assemble Fe-S clusters in a NifS-directed process, as monitored by the combination of UV-vis absorption, Mössbauer, resonance Raman, variable-temperature magnetic circular dichroism, and EPR spectroscopies. Although (Nif)IscA was found to bind ferrous ion in a tetrahedral, predominantly cysteinyl-ligated coordination environment, the low-binding affinity argues against a specific role as a metallochaperone for the delivery of ferrous ion to other Fe-S cluster assembly proteins. Rather, a role for (Nif)IscA as an alternate scaffold protein for Fe-S cluster biosynthesis is proposed, based on the NifS-directed assembly of approximately one labile [4Fe-4S](2+) cluster per (Nif)IscA homodimer, via a transient [2Fe-2S](2+) cluster intermediate. The cluster assembly process was monitored temporally using UV-vis absorption and Mössbauer spectroscopy, and the intermediate [2Fe-2S](2+)-containing species was additionally characterized by resonance Raman spectroscopy. The Mössbauer and resonance Raman properties of the [2Fe-2S](2+) center are consistent with complete cysteinyl ligation. The presence of three conserved cysteine residues in all IscA proteins and the observed cluster stoichiometry of approximately one [2Fe-2S](2+) or one [4Fe-4S](2+) per homodimer suggest that both cluster types are subunit bridging. In addition, (Nif)IscA was shown to couple delivery of iron and sulfur by using ferrous ion to reduce sulfane sulfur. The ability of Fe-S scaffold proteins to couple the delivery of these two toxic and reactive Fe-S cluster precursors is likely to

  11. Key players and their role during mitochondrial iron-sulfur cluster biosynthesis.

    PubMed

    Rawat, Swati; Stemmler, Timothy L

    2011-01-17

    Iron-sulfur clusters are multifaceted iron-containing cofactors coordinated and utilized by numerous proteins in nearly all biological systems. Fe-S-cluster-containing proteins help direct pathways essential for cell viability and participate in biological applications ranging from nucleotide biosynthesis and stability, protein translation, enzyme catalysis, and mitochondrial metabolism. Fe-S-containing proteins function by utilizing the unique electronic and chemical properties inherent in the Fe containing cofactor. Fe-S clusters are constructed of inorganic iron and sulfide arranged in a distinct caged structural makeup ranging from [Fe(2) -S(2) ], [Fe(3) -S(4) ], [Fe(4) -S(4) ], up to [Fe(8) -S(8) ] clusters. In eukaryotes, cluster activity is controlled in part at the assembly level and the major pathway for cluster production exists within the mitochondria. Recent insight into the pathway of mitochondrial cluster assembly has come from new in vivo and in vitro reports that provided direct insight into how all protein partners within the assembly pathway interact. However, we are only just beginning to understand the role of each protein within this complex pageant that is mitochondrial Fe-S cluster assembly. In this report we present results, using the yeast model for mitochondrial assembly, to describe the molecular details of how important proteins in the pathway coordinate for cluster assembly. PMID:21226084

  12. The electronic structure of the H-cluster in the [FeFe]-hydrogenase from Desulfovibrio desulfuricans: a Q-band 57Fe-ENDOR and HYSCORE study.

    PubMed

    Silakov, Alexey; Reijerse, Eduard J; Albracht, Simon P J; Hatchikian, E Claude; Lubitz, Wolfgang

    2007-09-19

    The active site of the (57)Fe-enriched [FeFe]-hydrogenase (i.e., the "H-cluster") from Desulfovibrio desulfuricans has been examined using advanced pulse EPR methods at X- and Q-band frequencies. For both the active oxidized state (H(ox)) and the CO inhibited form (H(ox)-CO) all six (57)Fe hyperfine couplings were detected. The analysis shows that the apparent spin density extends over the whole H-cluster. The investigations revealed different hyperfine couplings of all six (57)Fe nuclei in the H-cluster of the H(ox)-CO state. Four large 57Fe hyperfine couplings in the range 20-40 MHz were found (using pulse ENDOR and TRIPLE methods) and were assigned to the [4Fe-4S](H) (cubane) subcluster. Two weak (57)Fe hyperfine couplings below 5 MHz were identified using Q-band HYSCORE spectroscopy and were assigned to the [2Fe](H) subcluster. For the H(ox) state only two different 57Fe hyperfine couplings in the range 10-13 MHz were detected using pulse ENDOR. An (57)Fe line broadening analysis of the X-band CW EPR spectrum indicated, however, that all six (57)Fe nuclei in the H-cluster are contributing to the hyperfine pattern. It is concluded that in both states the binuclear subcluster [2Fe](H) assumes a [Fe(I)Fe(II)] redox configuration where the paramagnetic Fe(I) atom is attached to the [4Fe-4S](H) subcluster. The (57)Fe hyperfine interactions of the formally diamagnetic [4Fe-4S](H) are due to an exchange interaction between the two subclusters as has been discussed earlier by Popescu and Münck [Popescu, C.V.; Münck, E., J. Am. Chem. Soc. 1999, 121, 7877-7884]. This exchange coupling is strongly enhanced by binding of the extrinsic CO ligand. Binding of the dihydrogen substrate may induce a similar effect, and it is therefore proposed that the observed modulation of the electronic structure by the changing ligand surrounding plays an important role in the catalytic mechanism of [FeFe]-hydrogenase. PMID:17722921

  13. The resistance of electron-transport chain Fe-S clusters to oxidative damage during the reaction of peroxynitrite with mitochondrial complex II and rat-heart pericardium.

    PubMed

    Pearce, Linda L; Martinez-Bosch, Sandra; Manzano, Elisenda Lopez; Winnica, Daniel E; Epperly, Michael W; Peterson, Jim

    2009-05-01

    The effects of peroxynitrite and nitric oxide on the iron-sulfur clusters in complex II (succinate dehydrogenase) isolated from bovine heart have been studied primarily by EPR spectroscopy and no measurable damage to the constitutive 2Fe-2S, 3Fe-4S, or 4Fe-4S clusters was observed. The enzyme can be repeatedly oxidized with a slight excess of peroxynitrite and then quantitatively re-reduced with succinate. When added in large excess, peroxynitrite reacted with at least one tyrosine in each subunit of complex II to form 3-nitrotyrosines, but activity was barely compromised. Examination of rat-heart pericardium subjected to conditions leading to peroxynitrite production showed a small inhibition of complex II (16%) and a greater inhibition of aconitase (77%). In addition, experiments performed with excesses of sodium citrate and sodium succinate on rat-heart pericardium indicated that the "g approximately 2.01" EPR signal observed immediately following the beginning of conditions modeling oxidative/nitrosative stress, could be a consequence of both reversible oxidation of the constitutive 3Fe-4S cluster in complex II and degradation of the 4Fe-4S cluster in aconitase. However, the net signal envelope, which becomes apparent in less than 1min following the start of oxidative/nitrosative conditions, is dominated by the component arising from complex II. Taking into account the findings of a previous study concerning complexes I and III (L.L. Pearce, A.J. Kanai, M.W. Epperly, J. Peterson, Nitrosative stress results in irreversible inhibition of purified mitochondrial complexes I and III without modification of cofactors, Nitric Oxide 13 (2005) 254-263) it is now apparent that, with the exception of the cofactor in aconitase, mammalian (mitochondrial) iron-sulfur clusters are surprisingly resistant to degradation stemming from oxidative/nitrosative stress. PMID:19118636

  14. Nuclear resonance vibrational spectroscopy reveals the FeS cluster composition and active site vibrational properties of an O2-tolerant NAD+-reducing [NiFe] hydrogenase

    SciTech Connect

    Lauterbach, Lars; Wang, Hongxin; Horch, Marius; Gee, Leland B.; Yoda, Yoshitaka; Tanaka, Yoshihito; Zebger, Ingo; Lenz, Oliver; Cramer, Stephen P.

    2014-10-30

    Hydrogenases are complex metalloenzymes that catalyze the reversible splitting of molecular hydrogen into protons and electrons essentially without overpotential. The NAD+-reducing soluble hydrogenase (SH) from Ralstonia eutropha is capable of H2 conversion even in the presence of usually toxic dioxygen. The molecular details of the underlying reactions are largely unknown, mainly because of limited knowledge of the structure and function of the various metal cofactors present in the enzyme. Here, all iron-containing cofactors of the SH were investigated by 57Fe specific nuclear resonance vibrational spectroscopy (NRVS). Our data provide experimental evidence for one [2Fe2S] center and four [4Fe4S] clusters, which is consistent with the amino acid sequence composition. Only the [2Fe2S] cluster and one of the four [4Fe4S] clusters were reduced upon incubation of the SH with NADH. This finding explains the discrepancy between the large number of FeS clusters and the small amount of FeS cluster-related signals as detected by electron paramagnetic resonance spectroscopic analysis of several NAD+-reducing hydrogenases. For the first time, Fe–CO and Fe–CN modes derived from the [NiFe] active site could be distinguished by NRVS through selective 13C labeling of the CO ligand. This strategy also revealed the molecular coordinates that dominate the individual Fe–CO modes. The present approach explores the complex vibrational signature of the Fe–S clusters and the hydrogenase active site, thereby showing that NRVS represents a powerful tool for the elucidation of complex biocatalysts containing multiple cofactors.

  15. GPM Video of In-fa

    NASA Video Gallery

    On Nov. 23, GPM saw In-fa dropping rain at an extreme rate of over 266 mm (10.5 inches) per hour in storms just to the northwest of the typhoon's eye where thunderstorms reached altitudes of over 1...

  16. Characterization of iron-sulfur cluster assembly protein IscA from Acidithiobacillus ferrooxidans.

    PubMed

    Qian, Lin; Zheng, Chunli; Liu, Jianshe

    2013-03-01

    IscA is a key member of the iron-sulfur cluster assembly machinery found in bacteria and eukaryotes, but the mechanism of its function in the biogenesis of iron-sulfur cluster remains elusive. In this paper, we demonstrate that Acidithiobacillus ferrooxidans IscA is a [4Fe-4S] cluster binding protein, and it can bind iron in the presence of DTT with an apparent iron association constant of 4·10(20) M(-1). The iron binding in IscA can be promoted by oxygen through oxidizing ferrous iron to ferric iron. Furthermore, we show that the iron bound form of IscA can be converted to iron-sulfur cluster bound form in the presence of IscS and L-cysteine in vitro. Substitution of the invariant cysteine residues Cys35, Cys99, or Cys101 in IscA abolishes the iron binding activity of the protein; the IscA mutants that fail to bind iron are unable to assemble the iron-sulfur clusters. Further studies indicate that the iron-loaded IscA could act as an iron donor for the assembly of iron-sulfur clusters in the scaffold protein IscU in vitro. Taken together, these findings suggest that A. ferrooxidans IscA is not only an iron-sulfur protein, but also an iron binding protein that can act as an iron donor for biogenesis of iron-sulfur clusters. PMID:23586717

  17. Iron-sulfur cluster damage by the superoxide radical in neural tissues of the SOD1(G93A) ALS rat model.

    PubMed

    Popović-Bijelić, Ana; Mojović, Miloš; Stamenković, Stefan; Jovanović, Miloš; Selaković, Vesna; Andjus, Pavle; Bačić, Goran

    2016-07-01

    Extensive clinical investigations, in hand with biochemical and biophysical research, have associated brain iron accumulation with the pathogenesis of the amyotrophic lateral sclerosis (ALS) disease. The origin of iron is still not identified, but it is proposed that it forms redox active complexes that can participate in the Fenton reaction generating the toxic hydroxyl radical. In this paper, the state of iron in the neural tissues isolated from SOD1(G93A) transgenic rats was investigated using low temperature EPR spectroscopy and is compared with that of nontransgenic (NTg) littermates. The results showed that iron in neural tissues is present as high- and low-spin, heme and non-heme iron. It appears that the SOD1(G93A) rat neural tissues were most likely exposed in vivo to higher amounts of reactive oxygen species when compared to the corresponding NTg tissues, as they showed increased oxidized [3Fe-4S](1+) cluster content relative to [4Fe-4S](1+). Also, the activity of cytochrome c oxidase (CcO) was found to be reduced in these tissues, which may be associated with the observed uncoupling of heme a3 Fe and CuB in the O2-reduction site of the enzyme. Furthermore, the SOD1(G93A) rat spinal cords and brainstems contained more manganese, presumably from MnSOD, than those of NTg rats. The addition of potassium superoxide to all neural tissues ex vivo, led to the [4Fe-4S]→[3Fe-4S] cluster conversion and concurrent release of Fe. These results suggest that the superoxide anion may be the cause of the observed oxidative damage to SOD1(G93A) rat neural tissues and that the iron-sulfur clusters may be the source of poorly liganded redox active iron implicated in ALS pathogenesis. Low temperature EPR spectroscopy appears to be a valuable tool in assessing the role of metals in neurodegenerative diseases. PMID:27130034

  18. 2 CFR 200.414 - Indirect (F&A) costs.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... indirect costs. If an extension is granted the non-Federal entity may not request a rate review until the... REQUIREMENTS FOR FEDERAL AWARDS Cost Principles Direct and Indirect (f&a) Costs § 200.414 Indirect (F&A) costs... the types of cost which may be classified as indirect (F&A) cost in all situations....

  19. [FeFe]-hydrogenase oxygen inactivation is initiated at the H cluster 2Fe subcluster.

    PubMed

    Swanson, Kevin D; Ratzloff, Michael W; Mulder, David W; Artz, Jacob H; Ghose, Shourjo; Hoffman, Andrew; White, Spencer; Zadvornyy, Oleg A; Broderick, Joan B; Bothner, Brian; King, Paul W; Peters, John W

    2015-02-11

    The [FeFe]-hydrogenase catalytic site H cluster is a complex iron sulfur cofactor that is sensitive to oxygen (O2). The O2 sensitivity is a significant barrier for production of hydrogen as an energy source in water-splitting, oxygenic systems. Oxygen reacts directly with the H cluster, which results in rapid enzyme inactivation and eventual degradation. To investigate the progression of O2-dependent [FeFe]-hydrogenase inactivation and the process of H cluster degradation, the highly O2-sensitive [FeFe]-hydrogenase HydA1 from the green algae Chlamydomonas reinhardtii was exposed to defined concentrations of O2 while monitoring the loss of activity and accompanying changes in H cluster spectroscopic properties. The results indicate that H cluster degradation proceeds through a series of reactions, the extent of which depend on the initial enzyme reduction/oxidation state. The degradation process begins with O2 interacting and reacting with the 2Fe subcluster, leading to degradation of the 2Fe subcluster and leaving an inactive [4Fe-4S] subcluster state. This final inactive degradation product could be reactivated in vitro by incubation with 2Fe subcluster maturation machinery, specifically HydF(EG), which was observed by recovery of enzyme activity. PMID:25579778

  20. Two Fe-S clusters catalyse sulfur insertion by Radical-SAM methylthiotransferases

    PubMed Central

    Forouhar, Farhad; Arragain, Simon; Atta, Mohamed; Gambarelli, Serge; Mouesca, Jean-Marie; Hussain, Munif; Xiao, Rong; Kieffer-Jaquinod, Sylvie; Seetharaman, Jayaraman; Acton, Thomas B.; Montelione, Gaetano T.

    2014-01-01

    How living organisms create carbon-sulfur bonds during biosynthesis of critical sulphur-containing compounds is still poorly understood. The methylthiotransferases MiaB and RimO catalyze sulfur insertion into tRNAs and ribosomal protein S12, respectively. Both belong to a sub-group of Radical-SAM enzymes that bear two [4Fe-4S] clusters. One cluster binds S-Adenosylmethionine and generates an Ado• radical via a well- established mechanism. However, the precise role of the second cluster is unclear. For some sulfur-inserting Radical-SAM enzymes, this cluster has been proposed to act as a sacrificial source of sulfur for the reaction. In this paper, we report parallel enzymological, spectroscopic and crystallographic investigations of RimO and MiaB, which provide the first evidence that these enzymes are true catalysts and support a new sulfation mechanism involving activation of an exogenous sulfur co-substrate at an exchangeable coordination site on the second cluster, which remains intact during the reaction. PMID:23542644

  1. MAN or FA from n-butane

    SciTech Connect

    Di Cio, A.; Verde, L.

    1985-08-01

    Unsaturated polyester resins were first produced mostly from fumaric acid (FA) rather than from maleic anhydride (MAN). This is perfectly understandable if we consider that, using fumaric acid as raw material, polycondensates with a more homogeneous (less branched) structure are obtained, thus producing resins characterized by a more uniform and reproducible chemical and mechanical properties. Presently, for economical reasons, fumaric acid is used marginally as a MAN substitute in the production of polyester resins. These resins account for a major share (50%) of the overall MAN consumption in the U.S. and in Western Europe.

  2. Low-energy spectrum of iron-sulfur clusters directly from many-particle quantum mechanics

    NASA Astrophysics Data System (ADS)

    Sharma, Sandeep; Sivalingam, Kantharuban; Neese, Frank; Chan, Garnet Kin-Lic

    2014-10-01

    Iron-sulfur clusters are a universal biological motif. They carry out electron transfer, redox chemistry and even oxygen sensing, in diverse processes including nitrogen fixation, respiration and photosynthesis. Their low-lying electronic states are key to their remarkable reactivity, but they cannot be directly observed. Here, we present the first ever quantum calculation of the electronic levels of [2Fe-2S] and [4Fe-4S] clusters free from any model assumptions. Our results highlight the limitations of long-standing models of their electronic structure. In particular, we demonstrate that the widely used Heisenberg double exchange model underestimates the number of states by one to two orders of magnitude, which can conclusively be traced to the absence of Fe dd excitations, thought to be important in these clusters. Furthermore, the electronic energy levels of even the same spin are dense on the scale of vibrational fluctuations and this provides a natural explanation for the ubiquity of these clusters in catalysis in nature.

  3. Unanticipated coordination of tris buffer to the Radical SAM cluster of the RimO methylthiotransferase.

    PubMed

    Molle, Thibaut; Clémancey, Martin; Latour, Jean-Marc; Kathirvelu, Velavan; Sicoli, Giuseppe; Forouhar, Farhad; Mulliez, Etienne; Gambarelli, Serge; Atta, Mohamed

    2016-07-01

    Radical SAM enzymes generally contain a [4Fe-4S](2+/1+) (RS cluster) cluster bound to the protein via the three cysteines of a canonical motif CxxxCxxC. The non-cysteinyl iron is used to coordinate SAM via its amino-carboxylate moiety. The coordination-induced proximity between the cluster acting as an electron donor and the adenosyl-sulfonium bond of SAM allows for the homolytic cleavage of the latter leading to the formation of the reactive 5'-deoxyadenosyl radical used for substrate activation. Most of the structures of Radical SAM enzymes have been obtained in the presence of SAM, and therefore, little is known about the situation when SAM is not present. In this report, we show that RimO, a methylthiotransferase belonging to the radical SAM superfamily, binds a Tris molecule in the absence of SAM leading to specific spectroscopic signatures both in Mössbauer and pulsed EPR spectroscopies. These data provide a cautionary note for researchers who work with coordinative unsaturated iron sulfur clusters. PMID:27259294

  4. Fluorescent probes for tracking the transfer of iron-sulfur cluster and other metal cofactors in biosynthetic reaction pathways.

    PubMed

    Vranish, James N; Russell, William K; Yu, Lusa E; Cox, Rachael M; Russell, David H; Barondeau, David P

    2015-01-14

    Iron-sulfur (Fe-S) clusters are protein cofactors that are constructed and delivered to target proteins by elaborate biosynthetic machinery. Mechanistic insights into these processes have been limited by the lack of sensitive probes for tracking Fe-S cluster synthesis and transfer reactions. Here we present fusion protein- and intein-based fluorescent labeling strategies that can probe Fe-S cluster binding. The fluorescence is sensitive to different cluster types ([2Fe-2S] and [4Fe-4S] clusters), ligand environments ([2Fe-2S] clusters on Rieske, ferredoxin (Fdx), and glutaredoxin), and cluster oxidation states. The power of this approach is highlighted with an extreme example in which the kinetics of Fe-S cluster transfer reactions are monitored between two Fdx molecules that have identical Fe-S spectroscopic properties. This exchange reaction between labeled and unlabeled Fdx is catalyzed by dithiothreitol (DTT), a result that was confirmed by mass spectrometry. DTT likely functions in a ligand substitution reaction that generates a [2Fe-2S]-DTT species, which can transfer the cluster to either labeled or unlabeled Fdx. The ability to monitor this challenging cluster exchange reaction indicates that real-time Fe-S cluster incorporation can be tracked for a specific labeled protein in multicomponent assays that include several unlabeled Fe-S binding proteins or other chromophores. Such advanced kinetic experiments are required to untangle the intricate networks of transfer pathways and the factors affecting flux through branch points. High sensitivity and suitability with high-throughput methodology are additional benefits of this approach. We anticipate that this cluster detection methodology will transform the study of Fe-S cluster pathways and potentially other metal cofactor biosynthetic pathways. PMID:25478817

  5. Lipids and FA analysis of canine prostate tissue.

    PubMed

    Attar-Bashi, Nadia M; Orzeszko, Karyn; Slocombe, Ronald F; Sinclair, Andrew J

    2003-06-01

    It is widely reported that an association exists between dietary fat intake and the incidence of prostate cancer in humans. To study this association, there is a need for an animal model where prostate carcinogenesis occurs spontaneously. The canine prostate is considered a suitable experimental model for prostate cancer in humans since it is morphologically similar to the human prostate and both humans and dogs have a predisposition to benign and malignant prostate disease. In this study, the FA and lipids profiles of the normal canine prostate tissue from nine dogs were examined. The total lipid content of the canine prostate tissue was 1.7 +/- 0.5% (wet weight). The lipid composition analysis using TLC-FID showed that the two major lipid classes were phospholipids and TAG. Total FA, phospholipid, and TAG FA analysis showed that the major FA were palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), linoleic acid (18:2n-6), and arachidonic acid (20:4n-6). The n-3 FA were present at <3% of total FA and included alpha-linolenic acid (18:3n-3) (in total and TAG tissue FA), EPA (20:5n-3) (not in TAG), and DHA (22:6n-3) (not in TAG). The n-3/n-6 ratio was 1:11, 1:13, and 1:8 in total, phospholipid, and TAG FA, respectively. This study shows the canine prostate has a low level of n-3 FA and a low n-3/n-6 ratio. This is perhaps due to low n-3 content of the diet of the dogs. FA analysis of dogfoods available in Australia showed that the n-3 content in both supermarket and premium brand dogfoods was <3% (wet weight), and the n-3/n-6 ratio was low. PMID:12934677

  6. Time to Detection with BacT/Alert FA Plus Compared to BacT/Alert FA Blood Culture Media.

    PubMed

    Nutman, A; Fisher Even-Tsur, S; Shapiro, G; Braun, T; Schwartz, D; Carmeli, Y

    2016-09-01

    Rapid identification of the causative pathogen in patients with bacteremia allows adjustment of antibiotic therapy and improves patient outcomes. We compared in vitro and real-life time to detection (TTD) of two blood culture media, BacT/Alert FA (FA) and BacT/Alert FA Plus (FA Plus), for the nine most common species of bacterial pathogens recovered from blood samples. Experimental data from simulated cultures was compared with microbiology records of TTD for both culture media with growth of the species of interest in clinical blood cultures. In the experimental conditions, median TTD was 3.8 hours (23.9 %) shorter using FA Plus media. The magnitude of reduction differed between species. Similarly, in real life data, FA Plus had shorter TTD than FA media; however, the difference between culture media was smaller, and median TTD was only 1 hour (8.5 %) less. We found shorter TTD with BacT/Alert FA Plus culture media, both experimentally and in real-life conditions and unrelated to antibiotic neutralization, highlighting the importance of appropriate blood culture media selection. PMID:27272123

  7. Mass spectrometry of the lithium adducts of diacylglycerols containing hydroxy FA in castor oil and two normal FA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Castor oil can be used in industry. The molecular species of triacylglycerols containing hydroxy fatty acids (FA) in castor oil have been identified. We report here the identification of twelve diacylglycerols (DAG) containing hydroxy FA in castor oil using positive ion electrospray ionization mass ...

  8. Physical map of the chromosome of Neisseria gonorrhoeae FA1090 with locations of genetic markers, including opa and pil genes.

    PubMed Central

    Dempsey, J A; Litaker, W; Madhure, A; Snodgrass, T L; Cannon, J G

    1991-01-01

    A physical map of the chromosome of Neisseria gonorrhoeae FA1090 has been constructed. Digestion of strain FA1090 DNA with NheI, SpeI, BglII, or PacI resulted in a limited number of fragments that were resolved by contour-clamped homogeneous electric field electrophoresis. The estimated genome size was 2,219 kb. To construct the map, probes corresponding to single-copy chromosomal sequences were used in Southern blots of digested DNA separated on pulsed-field gels, to determine how the fragments from different digests overlapped. Some of the probes represented identified gonococcal genes, whereas others were anonymous cloned fragments of strain FA1090 DNA. By using this approach, a macrorestriction map of the strain FA1090 chromosome was assembled, and the locations of various genetic markers on the map were determined. Once the map was completed, the repeated gene families encoding Opa and pilin proteins were mapped. The 11 opa loci of strain FA1090 were distributed over approximately 60% of the chromosome. The pil loci were more clustered and were located in two regions separated by approximately one-fourth of the chromosome. Images PMID:1679431

  9. Epigenetics and development of food allergy (FA) in early childhood.

    PubMed

    Hong, Xiumei; Wang, Xiaobin

    2014-09-01

    This review aims to highlight the latest advance on epigenetics in the development of food allergy (FA) and to offer future perspectives. FA, a condition caused by an immunoglobulin (Ig) E-mediated hypersensitivity reaction to food, has emerged as a major clinical and public health problem worldwide in light of its increasing prevalence, potential fatality, and significant medical and economic impact. Current evidence supports that epigenetic mechanisms are involved in immune regulation and that the epigenome may represent a key "missing piece" of the etiological puzzle for FA. There are a growing number of population-based epigenetic studies on allergy-related phenotypes, mostly focused on DNA methylation. Previous studies mostly applied candidate-gene approaches and have demonstrated that epigenetic marks are associated with multiple allergic diseases and/or with early-life exposures relevant to allergy development (such as early-life smoking exposure, air pollution, farming environment, and dietary fat). Rapid technological advancements have made unbiased genome-wide DNA methylation studies highly feasible, although there are substantial challenge in study design, data analyses, and interpretation of findings. In conclusion, epigenetics represents both an important knowledge gap and a promising research area for FA. Due to the early onset of FA, epigenetic studies of FA in prospective birth cohorts have the potential to better understand gene-environment interactions and underlying biological mechanisms in FA during critical developmental windows (preconception, in utero, and early childhood) and may lead to new paradigms in the diagnosis, prevention, and management of FA and provide novel targets for future drug discovery and therapies for FA. PMID:25096861

  10. Hepatic denervation and dyslipidemia in obese Zucker (fa/fa) rats.

    PubMed

    Bruinstroop, E; Eliveld, J; Foppen, E; Busker, S; Ackermans, M T; Fliers, E; Kalsbeek, A

    2015-11-01

    Human and animal studies increasingly point toward a neural pathogenesis of the metabolic syndrome, involving hypothalamic and autonomic nervous system dysfunction. We hypothesized that increased very-low-density lipoprotein-triglyceride (VLDL-TG) secretion by the liver in a rat model for dyslipidemia, that is, the obese Zucker (fa/fa) rat, is due to relative hyperactivity of sympathetic, and/or hypoactivity of parasympathetic hepatic innervation. To test the involvement of the autonomic nervous system, we surgically denervated the sympathetic or parasympathetic hepatic nerve in obese Zucker rats. Our results show that cutting the sympathetic hepatic nerve lowers VLDL-TG secretion in obese rats, finally resulting in lower plasma TG concentrations after 6 weeks. In contrast, a parasympathetic denervation results in increased plasma total cholesterol concentrations. The effect of a sympathetic or parasympathetic denervation of the liver was independent of changes in humoral factors or changes in body weight or food intake. In conclusion, a sympathetic denervation improves the lipid profile in obese Zucker rats, whereas a parasympathetic denervation increases total cholesterol levels. We believe this is a novel treatment target, which should be further investigated. PMID:26134416

  11. The iron uptake repressor Fep1 in the fission yeast binds Fe-S cluster through conserved cysteines.

    PubMed

    Kim, Hyo-Jin; Lee, Kang-Lok; Kim, Kyoung-Dong; Roe, Jung-Hye

    2016-09-01

    Iron homeostasis is tightly regulated since iron is an essential but toxic element in the cell. The GATA-type transcription factor Fep1 and its orthologs contribute to iron homeostasis in many fungi by repressing genes for iron uptake when intracellular iron is high. Even though the function and interaction partners of Fep1 have been elucidated extensively In Schizosaccharomyces pombe, the mechanism behind iron-sensing by Fep1 remains elusive. It has been reported that Fep1 interacts with Fe-S-containing monothiol glutaredoxin Grx4 and Grx4-Fra2 complex. In this study, we demonstrate that Fep1 also binds iron, in the form of Fe-S cluster. Spectroscopic and biochemical analyses of as isolated and reconstituted Fep1 suggest that the dimeric Fep1 binds Fe-S clusters. The mutation study revealed that the cluster-binding depended on the conserved cysteines located between the two zinc fingers in the DNA binding domain. EPR analyses revealed [Fe-S]-specific peaks indicative of mixed presence of [2Fe-2S], [3Fe-4S], or [4Fe-4S]. The finding that Fep1 is an Fe-S protein fits nicely with the model that the Fe-S-trafficking Grx4 senses intracellular iron environment and modulates the activity of Fep1. PMID:27444384

  12. F/A-18 FAST Offers Advanced System Test Capability

    NASA Video Gallery

    NASA's Dryden Flight Research Center has modified an F/A-18A Hornet aircraft with additional research flight control computer systems for use as a Full-scale Advanced Systems Test Bed. Previously f...

  13. NASA Researches the 'FaINT' Side of Sonic Booms

    NASA Video Gallery

    As the latest in a continuing progression of NASA supersonics research projects aimed at reducing or mitigating the effect of sonic booms, the Farfield Investigation of No Boom Threshold, or FaINT,...

  14. F/A-18 Automated Aerial Refueling (AAR) Phase 1

    NASA Video Gallery

    Engineers at NASA's Dryden Flight Research Center are evaluating the capability of an F/A-18A aircraft as an in-flight refueling tanker to develop analytical models for an automated aerial refuelin...

  15. Xanthohumol lowers body weight and fasting plasma glucose in obese male Zucker fa/fa rats.

    PubMed

    Legette, Leecole L; Luna, Arlyn Y Moreno; Reed, Ralph L; Miranda, Cristobal L; Bobe, Gerd; Proteau, Rosita R; Stevens, Jan F

    2013-07-01

    Obesity contributes to increased risk for several chronic diseases including cardiovascular disease and type 2 diabetes. Xanthohumol, a prenylated flavonoid from hops (Humulus lupulus), was tested for efficacy on biomarkers of metabolic syndrome in 4 week old Zucker fa/fa rats, a rodent model of obesity. Rats received daily oral doses of xanthohumol at 0, 1.86, 5.64, and 16.9 mg/kg BW for 6 weeks. All rats were maintained on a high fat (60% kcal) AIN-93G diet for 3 weeks to induce severe obesity followed by a normal AIN-93G (15% kcal fat) diet for the last 3 weeks of the study. Weekly food intake and body weight were recorded. Plasma cholesterol, glucose, insulin, triglyceride, and monocyte chemoattractant protein-1 (MCP-1) levels were assessed using commercial assay kits. Plasma and liver tissue levels of XN and its metabolites were determined by liquid-chromatography tandem mass spectrometry. Plasma and liver tissue levels of xanthohumol were similar between low and medium dose groups and significantly (p<0.05) elevated in the highest dose group. There was a dose-dependent effect on body weight and plasma glucose levels. The highest dose group (n=6) had significantly lower plasma glucose levels compared to the control group (n=6) in male but not female rats. There was also a significant decrease in body weight for male rats in the highest dose group (16.9 mg/kg BW) compared to rats that received no xanthohumol, which was also not seen for female rats. Plasma cholesterol, insulin, triglycerides, and MCP-1 as well as food intake were not affected by treatment. The findings suggest that xanthohumol has beneficial effects on markers of metabolic syndrome. PMID:22640929

  16. Incorporation of [35S]sulfide into the Fe-S cluster of aconitase.

    PubMed

    Kennedy, M C; Emptage, M H; Beinert, H

    1984-03-10

    Exchange of sulfide into the iron-sulfur cluster of beef heart aconitase was investigated using Na235S. After anaerobic incubation for minutes and up to 24 h, samples were freed of substances of low Mr by Sephadex G-50 and analyzed for protein, protein-bound Fe, S2-, total Fe-S cluster by EPR, and for radioactivity. The activated and the inactive enzyme exchange three S2- ions within 1-2 h at essentially equal rates. No further exchange is observed with the activated enzyme within 24 h, whereas with the inactive enzyme, (partial) exchange of one more S2- occurs slowly within 5-10 h. Exchange is facilitated at elevated pH, but then destruction of clusters also increases. During incubation of inactive enzyme with S2-, partial activation may occur because S2- can act as reductant. Thus, depending on conditions, we observed very low to substantial activities. There is a linear, positive correlation between activity developed and ratio of cluster bound Fe to S2-. When Fe and dithiothreitol are present together with S2- during incubation, the extent of S2-exchange generally is between 10 and 25%. Fe incorporation exceeds S2- exchange, with the difference between Fe and S2- incorporation consistently amounting to one Fe/cluster. It is suggested that this excess Fe represents the Fe ion taken up on completion of the [4Fe-4S] from the [3Fe-4S] structure. The ease of S2-exchange suggests that the Fe-S cluster of aconitase is readily accessible to solvent. PMID:6699009

  17. Importance of Hydrogen Bonding in Fine Tuning the [2Fe-2S] Cluster Redox Potential of HydC from Thermotoga maritima.

    PubMed

    Birrell, James A; Laurich, Christoph; Reijerse, Edward J; Ogata, Hideaki; Lubitz, Wolfgang

    2016-08-01

    Iron-sulfur clusters form one of the largest and most diverse classes of enzyme cofactors in nature. They may serve as structural factors, form electron transfer chains between active sites and external redox partners, or form components of enzyme active sites. Their specific role is a consequence of the cluster type and the surrounding protein environment. The relative effects of these factors are not completely understood, and it is not yet possible to predict the properties of iron-sulfur clusters based on amino acid sequences or rationally tune their properties to generate proteins with new desirable functions. Here, we generate mutations in a [2Fe-2S] cluster protein, the TmHydC subunit of the trimeric [FeFe]-hydrogenase from Thermotoga maritima, to study the factors that affect its redox potential. Saturation mutagenesis of Val131 was used to tune the redox potential over a 135 mV range and revealed that cluster redox potential and electronic properties correlate with amino acid hydrophobicity and the ability to form hydrogen bonds to the cluster. Proline scanning mutagenesis between pairs of ligating cysteines was used to remove backbone amide hydrogen bonds to the cluster and decrease the redox potential by up to 132 mV, without large structural changes in most cases. However, substitution of Gly83 with proline caused a change of HydC to a [4Fe-4S] cluster protein with a redox potential of -526 mV. Together, these results confirm the importance of hydrogen bonding in tuning cluster redox potentials and demonstrate the versatility of iron-sulfur cluster protein folds at binding different types of clusters. PMID:27396836

  18. Characterization of two dissimilatory sulfite reductases from sulfate-reducing bacteria

    NASA Astrophysics Data System (ADS)

    Huynh, B. H.; Moura, I.; Lino, A. R.; Moura, J. J. G.; Legall, J.

    1988-02-01

    Mössbauer, EPR, and biochemical techniques were used to characterize two dissimilatory sulfite reductases: desulforubidin from Desulfovibrio baculatus strain DSM 1743 and desulfoviridin from Desulfovibrio gigas. For each molecule of desulforubidin, there are two sirohemes and four [4Fe-4S] clusters. The [4Fe-4S] clusters are in the diamagnetic 2+ oxidation state. The sirohemes are high-spin ferric (S=5/2) and each siroheme is exchanged-coupled to a [4Fe-4S]2+ cluster. Such an exchange-coupled siroheme-[4Fe-4S] unit has also been found in the assimilatory sulfite reductase from Escherichia coli/1/ and in a low-molecular weight sulfite reductase from Desulfovibrio vulgaris/2/. For each molecule of defulfoviridin, there are two tetrahydroporphyrin groups and four [4Fe-4S]2+ clusters. To our surprise, we discovered that about 80% of the tetrahydroporphyrin groups, however, do not bind iron.

  19. Dark-operative protochlorophyllide oxidoreductase generates substrate radicals by an iron-sulphur cluster in bacteriochlorophyll biosynthesis

    PubMed Central

    Nomata, Jiro; Kondo, Toru; Mizoguchi, Tadashi; Tamiaki, Hitoshi; Itoh, Shigeru; Fujita, Yuichi

    2014-01-01

    Photosynthesis converts solar energy to chemical energy using chlorophylls (Chls). In a late stage of biosynthesis of Chls, dark-operative protochlorophyllide (Pchlide) oxidoreductase (DPOR), a nitrogenase-like enzyme, reduces the C17 = C18 double bond of Pchlide and drastically changes the spectral properties suitable for photosynthesis forming the parental chlorin ring for Chl a. We previously proposed that the spatial arrangement of the proton donors determines the stereospecificity of the Pchlide reduction based on the recently resolved structure of the DPOR catalytic component, NB-protein. However, it was not clear how the two-electron and two-proton transfer events are coordinated in the reaction. In this study, we demonstrate that DPOR initiates a single electron transfer reaction from a [4Fe-4S]-cluster (NB-cluster) to Pchlide, generating Pchlide anion radicals followed by a single proton transfer, and then, further electron/proton transfer steps transform the anion radicals into chlorophyllide (Chlide). Thus, DPOR is a unique iron-sulphur enzyme to form substrate radicals followed by sequential proton- and electron-transfer steps with the protein folding very similar to that of nitrogenase. This novel radical-mediated reaction supports the biosynthesis of Chl in a wide variety of photosynthetic organisms. PMID:24965831

  20. "What D'ya Mean, Project SOL-FA?"

    ERIC Educational Resources Information Center

    Hardman, Olga S.

    1981-01-01

    Since 1977, Project Sol-fa, funded with an ESEA Title IV-C grant has provided Harrison County primary teachers with inservice training in the Kodaly method of music education. This article provides information on program funding, costs, and accomplishments. Evaluation forms and the inservice syllabus are appended. (SJL)

  1. IRAC F/A-18 RFCS on NASA 853

    NASA Technical Reports Server (NTRS)

    Pahli, Joseph

    2009-01-01

    F/A-18 testbed development and flight research are highlighted in this presentation. The current focus is on stability, specifically adaptive flight control, but soon the focus will move towards stability and maneuverability, examining flight planning and guidance, adaptive flight control, engine control and airframe and structures. Later research will additionally review V and V methods. Current and future IRAC plans are highlighted.

  2. F/A-18 tactical reconnaissance (tac recce) capability

    NASA Astrophysics Data System (ADS)

    Heinz, David; Pugh, Gregory G.; Wolters, David

    1996-11-01

    In 1995 the F/A-18 TAC RECCE Program was expanded beyond the initial electro-optic and infrared image recording capability of the Advanced Tactical Air Reconnaissance System (ATARS). The program now also includes integration of new high-resolution Synthetic Aperture Radar (SAR) reconnaissance modes and air-to-ground data link of ATARS and SAR image data. Delivery of the first reconnaissance equipment production units and fleet release is scheduled for 1998. F/A-18D two-seat aircraft will be retrofit with the RECCE equipment and designated F/A-18D(RC) (Reconnaissance Capable). This presentation describes recent F/A-18D(RC) operational assessment results, tactical reconnaissance equipment, functions, and interfaces for the 1998 fleet release. The equipment consists of the ATARS, the aircraft RECCE kit (access door, sensor windows, fairings), the upgraded APG-73 radar, and the data link pod. Functions include mission planning, automatic and manual acquisition of RECCE targets, image data recording, crew-station image review and edit, and data link. Interfaces include those with the Tactical Automated Mission Planning System, ground exploitation stations, and the aircraft carrier environment.

  3. Joining silicon nitride to FA-129 iron aluminide

    NASA Astrophysics Data System (ADS)

    Brochu, Mathieu

    Joints between dissimilar materials are characterized particularly by compositional gradients and microstructural changes, which yield large variations in chemical, physical and mechanical properties across the joint. The joining of dissimilar materials is therefore more complex than the joining of similar materials. In this project, the joining procedure, from the interaction between the different components in a joint to the determination of the mechanical properties was applied to the Si3N4/FA-129 system. This iron aluminide intermetallic alloy (FA-129), was developed by Oak Ridge National Laboratories (ORNL) to have high temperature properties with good room temperature ductility. This intermetallic is replacing high strength ferritic stainless steel (SS) in moderate strength applications due to cost and property reasons. Joints between SS and Si3N4 are already used industrially and this project was to evaluate the potential to replace these Si3N 4/SS joints by those of Si3N4/FA-129. Broadly stated, the results obtained during this project are as follows: (I) The E2 energy for Si3N4 ceramic was calculated to be 3.01 keV. (II) The wetting of iron aluminide alloy by copper has been achieved and the spreading and reaction kinetics are influenced by the presence of Cr as alloying element. (III) The penetration and decohesion of the FA-129 microstructure is significantly reduced by the utilization of a Cu alloy containing a high titanium concentration. (IV) An active brazing alloy containing a high active element content can be fabricated by an electroless deposition technique. (V) The melting behavior of the powder was characterized and complete melting occurs in a multi-step process at different temperatures, which are a function of the heating rate. (VI) The strength of joint produced by brazing Si3N4 to itself using the composite powder reached 400 MPa. (VII) Direct brazing of Si 3N4 to FA-129 was shown to be unsuccessful and therefore a soft Cu interlayer was

  4. A thermostable hybrid cluster protein from Pyrococcus furiosus: effects of the loss of a three helix bundle subdomain.

    PubMed

    Overeijnder, Marieke L; Hagen, Wilfred R; Hagedoorn, Peter-Leon

    2009-06-01

    Pyrococcus furiosus hybrid cluster protein (HCP) was expressed in Escherichia coli, purified, and characterized. This is the first archaeal and thermostable HCP to be isolated. Compared with the protein sequences of previously characterized HCPs from mesophiles, the protein sequence of P. furiosus HCP exhibits a deletion of approximately 13 kDa as a single amino acid stretch just after the N-terminal cysteine motif, characteristic for class-III HCPs from (hyper)thermophilic archaea and bacteria. The protein was expressed as a thermostable, soluble homodimeric protein. Hydroxylamine reductase activity of P. furiosus HCP showed a K(m) value of 0.40 mM and a k(cat) value of 3.8 s(-1) at 70 degrees C and pH 9.0. Electron paramagnetic resonance spectroscopy showed evidence for the presence of a spin-admixed, S = 3/2 [4Fe-4S](+) cubane cluster and of the hybrid cluster. The cubane cluster of P. furiosus HCP is presumably coordinated by a CXXC-X(7)-C-X(5)-C motif close to the N-terminus, which is similar to the CXXC-X(8)-C-X(5)-C motif of the Desulfovibrio desulfuricans and Desulfovibrio vulgaris HCPs. Amino acid sequence alignment and homology modeling of P. furiosus HCP reveal that the deletion results in a loss of one of the two three-helix bundles of domain 1. Clearly the loss of one of the three-helix bundles of domain 1 does not diminish the hydroxylamine reduction activity and the incorporation of the iron-sulfur clusters. PMID:19241093

  5. Electronic structure of the unique [4Fe-3S] cluster in O2-tolerant hydrogenases characterized by 57Fe Mossbauer and EPR spectroscopy.

    PubMed

    Pandelia, Maria-Eirini; Bykov, Dmytro; Izsak, Robert; Infossi, Pascale; Giudici-Orticoni, Marie-Thérèse; Bill, Eckhard; Neese, Frank; Lubitz, Wolfgang

    2013-01-01

    Iron-sulfur clusters are ubiquitous electron transfer cofactors in hydrogenases. Their types and redox properties are important for H(2) catalysis, but, recently, their role in a protection mechanism against oxidative inactivation has also been recognized for a [4Fe-3S] cluster in O(2)-tolerant group 1 [NiFe] hydrogenases. This cluster, which is uniquely coordinated by six cysteines, is situated in the proximity of the catalytic [NiFe] site and exhibits unusual redox versatility. The [4Fe-3S] cluster in hydrogenase (Hase) I from Aquifex aeolicus performs two redox transitions within a very small potential range, forming a superoxidized state above +200 mV vs. standard hydrogen electrode (SHE). Crystallographic data has revealed that this state is stabilized by the coordination of one of the iron atoms to a backbone nitrogen. Thus, the proximal [4Fe-3S] cluster undergoes redox-dependent changes to serve multiple purposes beyond classical electron transfer. In this paper, we present field-dependent (57)Fe-Mössbauer and EPR data for Hase I, which, in conjunction with spectroscopically calibrated density functional theory (DFT) calculations, reveal the distribution of Fe valences and spin-coupling schemes for the iron-sulfur clusters. The data demonstrate that the electronic structure of the [4Fe-3S] core in its three oxidation states closely resembles that of corresponding conventional [4Fe-4S] cubanes, albeit with distinct differences for some individual iron sites. The medial and distal iron-sulfur clusters have similar electronic properties as the corresponding cofactors in standard hydrogenases, although their redox potentials are higher. PMID:23267108

  6. North façade, entrance. The square tower has the remains of ...

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

    North façade, entrance. The square tower has the remains of a sign, Kaiser Foundation Hospital. Horizontal ribbon windows continue on this façade. - Richmond Field Hospital, 1330 Cutting Boulevard, Richmond, Contra Costa County, CA

  7. Central nervous system dysfunction in a mouse model of FA2H deficiency.

    PubMed

    Potter, Kathleen A; Kern, Michael J; Fullbright, George; Bielawski, Jacek; Scherer, Steven S; Yum, Sabrina W; Li, Jian J; Cheng, Hua; Han, Xianlin; Venkata, Jagadish Kummetha; Khan, P Akbar Ali; Rohrer, Bärbel; Hama, Hiroko

    2011-07-01

    Fatty acid 2-hydroxylase (FA2H) is responsible for the synthesis of myelin galactolipids containing hydroxy fatty acid (hFA) as the N-acyl chain. Mutations in the FA2H gene cause leukodystrophy, spastic paraplegia, and neurodegeneration with brain iron accumulation. Using the Cre-lox system, we developed two types of mouse mutants, Fa2h(-/-) mice (Fa2h deleted in all cells by germline deletion) and Fa2h(flox/flox) Cnp1-Cre mice (Fa2h deleted only in oligodendrocytes and Schwann cells). We found significant demyelination, profound axonal loss, and abnormally enlarged axons in the CNS of Fa2h(-/-) mice at 12 months of age, while structure and function of peripheral nerves were largely unaffected. Fa2h(-/-) mice also exhibited histological and functional disruption in the cerebellum at 12 months of age. In a time course study, significant deterioration of cerebellar function was first detected at 7 months of age. Further behavioral assessments in water T-maze and Morris water maze tasks revealed significant deficits in spatial learning and memory at 4 months of age. These data suggest that various regions of the CNS are functionally compromised in young adult Fa2h(-/-) mice. The cerebellar deficits in 12-month-old Fa2h(flox/flox) Cnp1-Cre mice were indistinguishable from Fa2h(-/-) mice, indicating that these phenotypes likely stem from the lack of myelin hFA-galactolipids. In contrast, Fa2h(flox/flox) Cnp1-Cre mice did not show reduced performance in water maze tasks, indicating that oligodendrocytes are not involved in the learning and memory deficits found in Fa2h(-/-) mice. These findings provide the first evidence that FA2H has an important function outside of oligodendrocytes in the CNS. PMID:21491498

  8. Opuntia ficus indica (nopal) attenuates hepatic steatosis and oxidative stress in obese Zucker (fa/fa) rats.

    PubMed

    Morán-Ramos, Sofía; Avila-Nava, Azalia; Tovar, Armando R; Pedraza-Chaverri, José; López-Romero, Patricia; Torres, Nimbe

    2012-11-01

    Nonalcoholic fatty liver disease (NAFLD) is associated with multiple factors such as obesity, insulin resistance, and oxidative stress. Nopal, a cactus plant widely consumed in the Mexican diet, is considered a functional food because of its antioxidant activity and ability to improve biomarkers of metabolic syndrome. The aim of this study was to assess the effect of nopal consumption on the development of hepatic steatosis and hepatic oxidative stress and on the regulation of genes involved in hepatic lipid metabolism. Obese Zucker (fa/fa) rats were fed a control diet or a diet containing 4% nopal for 7 wk. Rats fed the nopal-containing diet had ∼50% lower hepatic TG than the control group as well as a reduction in hepatomegaly and biomarkers of hepatocyte injury such as alanine and aspartate aminotransferases. Attenuation of hepatic steatosis by nopal consumption was accompanied by a higher serum concentration of adiponectin and a greater abundance of mRNA for genes involved in lipid oxidation and lipid export and production of carnitine palmitoyltransferase-1 and microsomal TG transfer proteins in liver. Hepatic reactive oxygen species and lipid peroxidation biomarkers were significantly lower in rats fed nopal compared with the control rats. Furthermore, rats fed the nopal diet had a lower postprandial serum insulin concentration and a greater liver phosphorylated protein kinase B (pAKT):AKT ratio in the postprandial state. This study suggests that nopal consumption attenuates hepatic steatosis by increasing fatty acid oxidation and VLDL synthesis, decreasing oxidative stress, and improving liver insulin signaling in obese Zucker (fa/fa) rats. PMID:23014486

  9. Ligand Binding to the FA3-FA4 Cleft Inhibits the Esterase-Like Activity of Human Serum Albumin

    PubMed Central

    Ascenzi, Paolo; Leboffe, Loris; di Masi, Alessandra; Trezza, Viviana; Fanali, Gabriella; Gioia, Magda; Coletta, Massimo; Fasano, Mauro

    2015-01-01

    The hydrolysis of 4-nitrophenyl esters of hexanoate (NphOHe) and decanoate (NphODe) by human serum albumin (HSA) at Tyr411, located at the FA3-FA4 site, has been investigated between pH 5.8 and 9.5, at 22.0°C. Values of Ks, k+2, and k+2/Ks obtained at [HSA] ≥ 5×[NphOXx] and [NphOXx] ≥ 5×[HSA] (Xx is NphOHe or NphODe) match very well each other; moreover, the deacylation step turns out to be the rate limiting step in catalysis (i.e., k+3 << k+2). The pH dependence of the kinetic parameters for the hydrolysis of NphOHe and NphODe can be described by the acidic pKa-shift of a single amino acid residue, which varies from 8.9 in the free HSA to 7.6 and 7.0 in the HSA:NphOHe and HSA:NphODe complex, respectively; the pK>a-shift appears to be correlated to the length of the fatty acid tail of the substrate. The inhibition of the HSA-Tyr411-catalyzed hydrolysis of NphOHe, NphODe, and 4-nitrophenyl myristate (NphOMy) by five inhibitors (i.e., diazepam, diflunisal, ibuprofen, 3-indoxyl-sulfate, and propofol) has been investigated at pH 7.5 and 22.0°C, resulting competitive. The affinity of diazepam, diflunisal, ibuprofen, 3-indoxyl-sulfate, and propofol for HSA reflects the selectivity of the FA3-FA4 cleft. Under conditions where Tyr411 is not acylated, the molar fraction of diazepam, diflunisal, ibuprofen, and 3-indoxyl-sulfate bound to HSA is higher than 0.9 whereas the molar fraction of propofol bound to HSA is ca. 0.5. PMID:25790235

  10. Ligand binding to the FA3-FA4 cleft inhibits the esterase-like activity of human serum albumin.

    PubMed

    Ascenzi, Paolo; Leboffe, Loris; di Masi, Alessandra; Trezza, Viviana; Fanali, Gabriella; Gioia, Magda; Coletta, Massimo; Fasano, Mauro

    2015-01-01

    The hydrolysis of 4-nitrophenyl esters of hexanoate (NphOHe) and decanoate (NphODe) by human serum albumin (HSA) at Tyr411, located at the FA3-FA4 site, has been investigated between pH 5.8 and 9.5, at 22.0°C. Values of Ks, k+2, and k+2/Ks obtained at [HSA] ≥ 5×[NphOXx] and [NphOXx] ≥ 5×[HSA] (Xx is NphOHe or NphODe) match very well each other; moreover, the deacylation step turns out to be the rate limiting step in catalysis (i.e., k+3 < k+2). The pH dependence of the kinetic parameters for the hydrolysis of NphOHe and NphODe can be described by the acidic pKa-shift of a single amino acid residue, which varies from 8.9 in the free HSA to 7.6 and 7.0 in the HSA:NphOHe and HSA:NphODe complex, respectively; the pK>a-shift appears to be correlated to the length of the fatty acid tail of the substrate. The inhibition of the HSA-Tyr411-catalyzed hydrolysis of NphOHe, NphODe, and 4-nitrophenyl myristate (NphOMy) by five inhibitors (i.e., diazepam, diflunisal, ibuprofen, 3-indoxyl-sulfate, and propofol) has been investigated at pH 7.5 and 22.0°C, resulting competitive. The affinity of diazepam, diflunisal, ibuprofen, 3-indoxyl-sulfate, and propofol for HSA reflects the selectivity of the FA3-FA4 cleft. Under conditions where Tyr411 is not acylated, the molar fraction of diazepam, diflunisal, ibuprofen, and 3-indoxyl-sulfate bound to HSA is higher than 0.9 whereas the molar fraction of propofol bound to HSA is ca. 0.5. PMID:25790235

  11. This NASA Dryden F/A-18 is participating in the Automated Aerial Refueling (AAR) project. F/A-18 (No

    NASA Technical Reports Server (NTRS)

    2002-01-01

    A NASA Dryden F/A-18 is participating in the Automated Aerial Refueling (AAR) project. F/A-18 (No. 847) is acting as an in-flight refueling tanker in the study to develop analytical models for an automated aerial refueling system for unmanned vehicles. A 300-gallon aerodynamic pod containing air-refueling equipment is seen beneath the fuselage. The hose and refueling basket are extended during an assessment of their dynamics on the F/A-18A.

  12. FaPOD27 functions in the metabolism of polyphenols in strawberry fruit (Fragaria sp.)

    PubMed Central

    Yeh, Su-Ying; Huang, Fong-Chin; Hoffmann, Thomas; Mayershofer, Mechthild; Schwab, Wilfried

    2014-01-01

    The strawberry (Fragaria × ananassa) is one of the most preferred fresh fruit worldwide, accumulates numerous flavonoids but has limited shelf life due to excessive tissue softening caused by cell wall degradation. Since lignin is one of the polymers that strengthen plant cell walls and might contribute to some extent to fruit firmness monolignol biosynthesis was studied in strawberry fruit. Cinnamoyl-CoA reductase (CCR), cinnamyl alcohol dehydrogenase (CAD), and a peroxidase (POD27) gene were strongly expressed in red, ripe fruit whereas a second POD gene was primarily expressed in green, immature fruit. Moreover, FaPOD27 transcripts were strongly and constitutively induced in fruits exposed to Agrobacterium infection. Gene expression levels and enzymatic activities of FaCCR and FaCAD were efficiently suppressed through RNAi in FaCCR- and FaCAD-silenced strawberries. Besides, significantly elevated FaPOD transcript levels were detected after agroinfiltration of pBI-FaPOD constructs in fruits. At the same time, levels of G-monomers were considerably reduced in FaCCR-silenced fruits whereas the proportion of both G- and S-monomers decisively decreased in FaCAD-silenced and pBI-FaPOD fruits. Development, firmness, and lignin level of the treated fruits were similar to pBI-intron control fruits, presumably attributed to increased expression levels of FaPOD27 upon agroinfiltration. Additionally, enhanced firmness, accompanied with elevated lignin levels, was revealed in chalcone synthase-deficient fruits (CHS−), independent of down- or up-regulation of individual and combined FaCCR. FaCAD, and FaPOD genes by agroinfiltration, when compared to CHS−/pBI-intron control fruits. These approaches provide further insight into the genetic control of flavonoid and lignin synthesis in strawberries. The results suggest that FaPOD27 is a key gene for lignin biosynthesis in strawberry fruit and thus to improving the firmness of strawberries. PMID:25346738

  13. Characteristics of F/A-18 vertical tail buffeting

    NASA Astrophysics Data System (ADS)

    Sheta, E. F.; Huttsell, L. J.

    2003-03-01

    A time-accurate computational analysis of vertical tail buffeting of full F/A-18 aircraft is conducted at typical flight conditions to identify the buffet characteristics of fighter aircraft. The F/A-18 aircraft is pitched at wide range of high angles of attack at Mach number of 0.243 and Reynolds number of 11 millions. Strong coupling between the fluid and structure is considered in this investigation. Strong coupling occurs when the inertial effect of the motion of the vertical tail is fed back into the flow field. The aerodynamic flow field around the F/A-18 aircraft is computed using the Reynolds-averaged full Navier-Stokes equations. The dynamical structural response of the vertical tail is predicted using direct finite-element analysis. The interface between the fluid and structure is applied using conservative and consistent interfacing methodology. The motion of the computational grid due to the deflection of the vertical tail is computed using transfinite interpolation module. The investigation revealed that the vertical tail is subject to bending and torsional responses, mainly in the first modes of vibrations. The buffet loads increase significantly as the onset of vortex breakdown moves upstream of the vertical tails. The inboard surface of the vertical tail has more significant contribution in the buffet excitation than the outboard surface. In addition, the pressure on the outboard surface of the vertical tail is less sensitive to the angle of attack than the pressure on the inboard surface. The buffet excitation peaks shift to lower frequency as the angle of attack increases. The computational results are compared, and they are in close agreement, with several flight and experimental data.

  14. Isolation and Characterization of Frankia sp. Strain FaC1 Genes Involved in Nitrogen Fixation.

    PubMed

    Ligon, J M; Nakas, J P

    1987-10-01

    Genomic DNA was isolated from Frankia sp. strain FaC1, an Alnus root nodule endophyte, and used to construct a genomic library in the cosmid vector pHC79. The genomic library was screened by in situ colony hybridization to identify clones of Frankia nitrogenase (nif) genes based on DNA sequence homology to structural nitrogenase genes from Klebsiella pneumoniae. Several Frankia nif clones were isolated, and hybridization with individual structural nitrogenase gene fragments (nifH, nifD, and nifK) from K. pneumoniae revealed that they all contain the nifD and nifK genes, but lack the nifH gene. Restriction endonuclease mapping of the nifD and nifK hybridizing region from one clone revealed that the nifD and nifK genes in Frankia sp. are contiguous, while the nifH gene is absent from a large region of DNA on either side of the nifDK gene cluster. Additional hybridizations with gene fragments derived from K. pneumoniae as probes and containing other genes involved in nitrogen fixation demonstrated that the Frankia nifE and nifN genes, which play a role in the biosynthesis of the iron-molybdenum cofactor, are located adjacent to the nifDK gene cluster. PMID:16347453

  15. The dam replacing gene product enhances Neisseria gonorrhoeae FA1090 viability and biofilm formation

    PubMed Central

    Kwiatek, Agnieszka; Bacal, Pawel; Wasiluk, Adrian; Trybunko, Anastasiya; Adamczyk-Poplawska, Monika

    2014-01-01

    Many Neisseriaceae do not exhibit Dam methyltransferase activity and, instead of the dam gene, possess drg (dam replacing gene) inserted in the leuS/dam locus. The drg locus in Neisseria gonorrhoeae FA1090 has a lower GC-pairs content (40.5%) compared to the whole genome of N. gonorrhoeae FA1090 (52%). The gonococcal drg gene encodes a DNA endonuclease Drg, with GmeATC specificity. Disruption of drg or insertion of the dam gene in gonococcal genome changes the level of expression of genes as shown by transcriptome analysis. For the drg-deficient N. gonorrhoeae mutant, a total of 195 (8.94% of the total gene pool) genes exhibited an altered expression compared to the wt strain by at least 1.5 fold. In dam-expressing N. gonorrhoeae mutant, the expression of 240 genes (11% of total genes) was deregulated. Most of these deregulated genes were involved in translation, DNA repair, membrane biogenesis and energy production as shown by cluster of orthologous group analysis. In vivo, the inactivation of drg gene causes the decrease of the number of live neisserial cells and long lag phase of growth. The insertion of dam gene instead of drg locus restores cell viability. We have also shown that presence of the drg gene product is important for N. gonorrhoeae FA1090 in adhesion, including human epithelial cells, and biofilm formation. Biofilm produced by drg-deficient strain is formed by more dispersed cells, compared to this one formed by parental strain as shown by scanning electron and confocal microscopy. Also adherence assays show a significantly smaller biomass of formed biofilm (OD570 = 0.242 ± 0.038) for drg-deficient strain, compared to wild-type strain (OD570 = 0.378 ± 0.057). Dam-expressing gonococcal cells produce slightly weaker biofilm with cells embedded in an extracellular matrix. This strain has also a five times reduced ability for adhesion to human epithelial cells. In this context, the presence of Drg is more advantageous for N. gonorrhoeae biology than

  16. Biochemical and spectroscopic studies of epoxyqueuosine reductase: A novel iron-sulfur cluster and cobalamin containing protein involved in the biosynthesis of queuosine

    PubMed Central

    Miles, Zachary D.; Myers, William K.; Kincannon, William M.; Britt, R. David; Bandarian, Vahe

    2015-01-01

    Queuosine is a hypermodified nucleoside present in the wobble position of tRNAs with a 5′-GUN-3′ sequence in their anticodon (His, Asp, Asn, and Tyr). The 7-deazapurine core of the base is synthesized de novo in prokaryotes from guanosine-5′-triphosphate in a series of eight sequential enzymatic transformations, the final three occurring on tRNA. Epoxyqueuosine reductase (QueG), catalyzes the final step in the pathway, which entails the 2-electron reduction of epoxyqueuosine to form queuosine. Biochemical analyses reveal that this enzyme requires cobalamin and two [4Fe-4S] clusters for catalysis. Spectroscopic studies show that the cobalamin appears to bind in a base-off conformation, whereby the dimethylbenzimidazole moiety of the cofactor is removed from the coordination sphere of the cobalt but not replaced by an imidazole sidechain, which is a hallmark of many cobalamin-dependent enzymes. The bioinformatically-identified residues are shown to have a role in modulating the primary coordination sphere of cobalamin. These studies provide the first demonstration of the cofactor requirements for QueG. PMID:26230193

  17. Crystal structure of a prokaryotic (6-4) photolyase with an Fe-S cluster and a 6,7-dimethyl-8-ribityllumazine antenna chromophore

    PubMed Central

    Zhang, Fan; Scheerer, Patrick; Oberpichler, Inga; Lamparter, Tilman; Krauß, Norbert

    2013-01-01

    The (6-4) photolyases use blue light to reverse UV-induced (6-4) photoproducts in DNA. This (6-4) photorepair was thought to be restricted to eukaryotes. Here we report a prokaryotic (6-4) photolyase, PhrB from Agrobacterium tumefaciens, and propose that (6-4) photolyases are broadly distributed in prokaryotes. The crystal structure of photolyase related protein B (PhrB) at 1.45 Å resolution suggests a DNA binding mode different from that of the eukaryotic counterparts. A His-His-X-X-Arg motif is located within the proposed DNA lesion contact site of PhrB. This motif is structurally conserved in eukaryotic (6-4) photolyases for which the second His is essential for the (6-4) photolyase function. The PhrB structure contains 6,7-dimethyl-8-ribityllumazine as an antenna chromophore and a [4Fe-4S] cluster bound to the catalytic domain. A significant part of the Fe-S fold strikingly resembles that of the large subunit of eukaryotic and archaeal primases, suggesting that the PhrB-like photolyases branched at the base of the evolution of the cryptochrome/photolyase family. Our study presents a unique prokaryotic (6-4) photolyase and proposes that the prokaryotic (6-4) photolyases are the ancestors of the cryptochrome/photolyase family. PMID:23589886

  18. Basic firefly algorithm for document clustering

    NASA Astrophysics Data System (ADS)

    Mohammed, Athraa Jasim; Yusof, Yuhanis; Husni, Husniza

    2015-12-01

    The Document clustering plays significant role in Information Retrieval (IR) where it organizes documents prior to the retrieval process. To date, various clustering algorithms have been proposed and this includes the K-means and Particle Swarm Optimization. Even though these algorithms have been widely applied in many disciplines due to its simplicity, such an approach tends to be trapped in a local minimum during its search for an optimal solution. To address the shortcoming, this paper proposes a Basic Firefly (Basic FA) algorithm to cluster text documents. The algorithm employs the Average Distance to Document Centroid (ADDC) as the objective function of the search. Experiments utilizing the proposed algorithm were conducted on the 20Newsgroups benchmark dataset. Results demonstrate that the Basic FA generates a more robust and compact clusters than the ones produced by K-means and Particle Swarm Optimization (PSO).

  19. Depositional Environment of Permian Tak Fa Formation, Nakhonsawan, Northern Thailand

    NASA Astrophysics Data System (ADS)

    Ketwetsuriya, Chatchalerm; Nützel, Alexander; Kanjanapayont, Pitsanupong

    2016-04-01

    The carbonate rocks of the study area at Amphoe Tak Fa and Amphoe Takhli, Changwat Nakhon Sawan belong to the Tak Fa Formation, Saraburi Group. This formation crops out in the Khao Khwang Platform and consists of late Palaeozoic carbonate platform deposits. It reaches a thickness of 900 meters and crops out in a vast area. The exposures have been measured and samples were collected for petrographic study. The rock consists of limestones, argillaceous limestones, mudstones and dolomites with nodular and banded cherts, which comprise many invertebrate fossils such as fusulinids, ammonoid, pelecypod, gastropod, coral and bryozoa. Many of the fossils are silicified. The gastropod assemblage is currently under study and represents one of the most diverse faunas reported from SE Asia. The age of the rock is Yakhtashian or Artinskian (late Early Permian) to Midian or Capitanian (late Middle Permian). The study of carbonate facies and fauna indicates that the depositional environment was on shelf lagoon within the carbonate platform varying from shallow marine to barrier bar.

  20. Occurrence of Far-Red Light Photoacclimation (FaRLiP) in Diverse Cyanobacteria

    PubMed Central

    Gan, Fei; Shen, Gaozhong; Bryant, Donald A.

    2014-01-01

    Cyanobacteria have evolved a number of acclimation strategies to sense and respond to changing nutrient and light conditions. Leptolyngbya sp. JSC-1 was recently shown to photoacclimate to far-red light by extensively remodeling its photosystem (PS) I, PS II and phycobilisome complexes, thereby gaining the ability to grow in far-red light. A 21-gene photosynthetic gene cluster (rfpA/B/C, apcA2/B2/D2/E2/D3, psbA3/D3/C2/B2/H2/A4, psaA2/B2/L2/I2/F2/J2) that is specifically expressed in far-red light encodes the core subunits of the three major photosynthetic complexes. The growth responses to far-red light were studied here for five additional cyanobacterial strains, each of which has a gene cluster similar to that in Leptolyngbya sp. JSC-1. After acclimation all five strains could grow continuously in far-red light. Under these growth conditions each strain synthesizes chlorophylls d, f and a after photoacclimation, and each strain produces modified forms of PS I, PS II (and phycobiliproteins) that absorb light between 700 and 800 nm. We conclude that these photosynthetic gene clusters are diagnostic of the capacity to photoacclimate to and grow in far-red light. Given the diversity of terrestrial environments from which these cyanobacteria were isolated, it is likely that FaRLiP plays an important role in optimizing photosynthesis in terrestrial environments. PMID:25551681

  1. Solidification behavior of FA-129 iron-aluminide alloy

    SciTech Connect

    Viswanathan, S.; Maziasz, P.J.; Sikka, V.K.

    1992-07-01

    Samples of FA-129 iron-aluminide alloy have been processed by various casting processes, including vacuum arc-melting followed by chill casting into water-cooled copper molds, air melting followed by casting into graphite molds, and electroslag melting and casting into large ingots. The resulting microstructures obtained in these processes are examined and compared. Selected samples are homogenized at various temperatures and times. Optical and transmission electron microscopy (TEM) and energy dispersive spectroscopy (EDS) are used to identify the morphology, constituents, and compositions of various phases. Interrupted solidification experiments are used to study the evolution of cast structure. Some observations are made on the effect of dissolved hydrogen in the melt on as-cast soundness.

  2. Solidification behavior of FA-129 iron-aluminide alloy

    SciTech Connect

    Viswanathan, S.; Maziasz, P.J.; Sikka, V.K.

    1992-01-01

    Samples of FA-129 iron-aluminide alloy have been processed by various casting processes, including vacuum arc-melting followed by chill casting into water-cooled copper molds, air melting followed by casting into graphite molds, and electroslag melting and casting into large ingots. The resulting microstructures obtained in these processes are examined and compared. Selected samples are homogenized at various temperatures and times. Optical and transmission electron microscopy (TEM) and energy dispersive spectroscopy (EDS) are used to identify the morphology, constituents, and compositions of various phases. Interrupted solidification experiments are used to study the evolution of cast structure. Some observations are made on the effect of dissolved hydrogen in the melt on as-cast soundness.

  3. Involvement of fertilization antigen (FA-1) in involuntary immunoinfertility in humans.

    PubMed Central

    Naz, R K

    1987-01-01

    Sera from immunoinfertile patients (n = 32) and fertile controls (n = 20) were analyzed for cross-reaction with a purified and characterized sperm-specific glycoprotein, the fertilization antigen (FA-1), employing an enzyme-linked immunosorbent assay. The immunoinfertile sera demonstrated a strong reaction with FA-1 when compared with fertile control sera. There was no correlation between the reaction of sera with FA-1 and the titers obtained through the sperm agglutination technique and the sperm immobilization technique. Immunoinfertile sera showed binding with the protein bands in the regions corresponding to FA-1 on Western blots involving sodium deoxycholate-solubilized human sperm. Antigens isolated with immunoaffinity chromatography involving immunoinfertile sera also demonstrated antigen bands corresponding to FA-1 in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Of the seven immunoinfertile couples, three that had antibodies to FA-1 in the male as well as female partners demonstrated a block of fertilization (IVF) due to antibodies bound on the sperm surface. The anti-FA-1 antibody activity was detected in serum as well as in follicular fluid and seminal plasma. Immunoinfertile sera that showed an inhibition of human sperm penetration of zona-free hamster ova showed a significant (P less than 0.001) increase in penetration rates after absorption with FA-1. These results indicate that sera from immunoinfertile patients had antibodies reacting with FA-1, and these antibodies are involved in the fertilization process. Images PMID:3316276

  4. Yamadazyma insecticola f.a., sp. nov. and Yamadazyma epiphylla f.a., sp. nov., two novel yeast species.

    PubMed

    Jindamorakot, Sasitorn; Am-In, Somjit; Kaewwichian, Rungluk; Limtong, Savitree

    2015-04-01

    Two yeast strains representing two novel yeast species were isolated from frass of an unidentified insect (ST-78(T)) and the external surfaces of rice leaves (YE170(T)) collected in Thailand. The two strains were genetically, morphologically and phenotypically distinct from recognized species and were found to represent two novel species of the genus Yamadazyma although formation of ascospores was not observed. In terms of pairwise sequence similarity of the D1/D2 region of the large subunit rRNA gene, the closest relative of strain ST-78(T) was Candida lessepsii CBS 9941(T) but with 3.8% nucleotide substitutions, while the closest relative of strain YE170(T) was strain ST-78(T) but with 4.3% nucleotide substitutions. Analysis of the sequences of the internal transcribed spacer 1 and 2 (ITS1-2) regions revealed that strain ST-78 differed from C. lessepsii CBS 9941(T) by 8.8% nucleotide substitutions and from strain YE170(T) by 9.4% nucleotide substitutions. The result of pairwise sequence similarity of the D1/D2 and ITS1-2 regions together with phylogenetic analysis indicated that strains ST-78(T) and YE170(T) represented two novel species within the Yamadazyma clade. The names Yamadazyma insecticola f.a., sp. nov. (type strain ST-78(T) = BCC 8314(T) = NBRC 110421(T) = CBS 13382(T); MycoBank no. MB810546) and Yamadazyma epiphylla f.a., sp. nov. (type strain YE170(T) = BCC 63466(T) = NBRC 110423(T) = CBS 13384(T); MycoBank no. MB810547) are proposed for the two novel yeast species. PMID:25644483

  5. Phthalate catabolic gene cluster is linked to the angular dioxygenase gene in Terrabacter sp. strain DBF63.

    PubMed

    Habe, H; Miyakoshi, M; Chung, J; Kasuga, K; Yoshida, T; Nojiri, H; Omori, T

    2003-03-01

    Phthalate is a metabolic intermediate of the pathway of fluorene (FN) degradation via angular dioxygenation. A gene cluster responsible for the conversion of phthalate to protocatechuate was cloned from the dibenzofuran (DF)- and FN-degrading bacterium Terrabacter sp. strain DBF63 and sequenced. The genes encoding seven catabolic enzymes, oxygenase large subunit of phthalate 3,4-dioxygenase (phtA1), oxygenase small subunit of phthalate 3,4-dioxygenase (phtA2), cis-3,4-dihydroxy-3,4-dihydrophthalate dehydrogenase (phtB), [3Fe-4S] or [4Fe-4S] type of ferredoxin (phtA3), ferredoxin reductase (phtA4), 3,4-dihydroxyphthalate decarboxylase (phtC) and putative regulatory protein (phtR), were found in the upstream region of the angular dioxygenase gene (dbfA1A2), encoded in this order. Escherichia coli carrying phtA1A2BA3A4 genes converted phthalate to 3,4-dihydroxyphthalate, and the 3,4-dihydroxyphthalate decarboxylase activity by E. coli cells carrying phtC was finally detected with the introduction of a Shine-Dalgarno sequence in the upstream region of its initiation codon. Homology analysis on the upstream region of the pht gene cluster revealed that there was an insertion sequence (IS) (ISTesp2; ORF14 and its flanking region), part of which was almost 100% identical to the orf1 and its flanking region adjacent to the extradiol dioxygenase gene ( bphC1) involved in the DF degradation of Terrabacter sp. strain DPO360 [Schmid et al. (1997) J Bacteriol 179:53-62]. This suggests that ISTesp2 plays a role in the metabolism of aromatic compounds in Terrabacter sp. strains DBF63 and DPO360. PMID:12658514

  6. RfpA, RfpB, and RfpC are the Master Control Elements of Far-Red Light Photoacclimation (FaRLiP)

    PubMed Central

    Zhao, Chi; Gan, Fei; Shen, Gaozhong; Bryant, Donald A.

    2015-01-01

    Terrestrial cyanobacteria often occur in niches that are strongly enriched in far-red light (FRL; λ > 700 nm). Some cyanobacteria exhibit a complex and extensive photoacclimation response, known as FRL photoacclimation (FaRLiP). During the FaRLiP response, specialized paralogous proteins replace 17 core subunits of the three major photosynthetic complexes: Photosystem (PS) I, PS II, and the phycobilisome. Additionally, the cells synthesize both chlorophyll (Chl) f and Chl d. Using biparental mating from Escherichia coli, we constructed null mutants of three genes, rfpA, rfpB, and rfpC, in the cyanobacteria Chlorogloeopsis fritschii PCC 9212 and Chroococcidiopsis thermalis PCC 7203. The resulting mutants were no longer able to modify their photosynthetic apparatus to absorb FRL, were no longer able to synthesize Chl f, inappropriately synthesized Chl d in white light, and were unable to transcribe genes of the FaRLiP gene cluster. We conclude that RfpA, RfpB, and RfpC constitute a FRL-activated signal transduction cascade that is the master control switch for the FaRLiP response. FRL is proposed to activate (or inactivate) the histidine kinase activity of RfpA, which leads to formation of the active state of RfpB, the key response regulator and transcription activator. RfpC may act as a phosphate shuttle between RfpA and RfpB. Our results show that reverse genetics via conjugation will be a powerful approach in detailed studies of the FaRLiP response. PMID:26635768

  7. RfpA, RfpB, and RfpC are the master control elements of far-red light photoacclimation (FaRLiP)

    DOE PAGESBeta

    Zhao, Chi; Gan, Fei; Shen, Gaozhong; Bryant, Donald A.

    2015-11-25

    Terrestrial cyanobacteria often occur in niches tha tare strongly enriched in far-redlight (FRL; λ > 700nm). Some cyanobacteria exhibit a complex and extensive photoacclimation response, known as FRLphotoacclimation(FaRLiP).During the FaRLiP response, specialized paralogous proteins replace 17 core subunits of the three major photosynthetic complexes: Photosystem (PS)I, PSII,and the phycobilisome. Additionally, the cells synthesize both chlorophyll (Chl) f and Chl d.Using biparental mating from Escherichia coli, we constructed null mutants of three genes, rfpA, rfpB,and rfpC, in the cyanobacteria Chlorogloeopsis fritschii PCC 9212 and Chroococcidiopsis thermalis PCC 7203.The resulting mutants were no longer able to modify their photosynthetic apparatus to absorbmore » FRL, were no longer able to synthesize Chl f, in appropriately synthesized Chl d in white light,and were unable to transcribe genes of the FaRLiP gene cluster. We conclude that RfpA, RfpB, and RfpC constitute a FRL-activated signal transduction cascade that is the master control switch for the FaRLiP response. FRL is proposed to activate (or inactivate) the histidine kinase activity of RfpA, which leads to formation of the active state of RfpB, the key response regulator and transcription activator. RfpC may act as a phosphate shuttle between RfpA and RfpB. Our results show that reverse genetics via conjugation will be a powerful approach in detailed studies of the FaRLiP response.« less

  8. RfpA, RfpB, and RfpC are the master control elements of far-red light photoacclimation (FaRLiP)

    SciTech Connect

    Zhao, Chi; Gan, Fei; Shen, Gaozhong; Bryant, Donald A.

    2015-11-25

    Terrestrial cyanobacteria often occur in niches tha tare strongly enriched in far-redlight (FRL; λ > 700nm). Some cyanobacteria exhibit a complex and extensive photoacclimation response, known as FRLphotoacclimation(FaRLiP).During the FaRLiP response, specialized paralogous proteins replace 17 core subunits of the three major photosynthetic complexes: Photosystem (PS)I, PSII,and the phycobilisome. Additionally, the cells synthesize both chlorophyll (Chl) f and Chl d.Using biparental mating from Escherichia coli, we constructed null mutants of three genes, rfpA, rfpB,and rfpC, in the cyanobacteria Chlorogloeopsis fritschii PCC 9212 and Chroococcidiopsis thermalis PCC 7203.The resulting mutants were no longer able to modify their photosynthetic apparatus to absorb FRL, were no longer able to synthesize Chl f, in appropriately synthesized Chl d in white light,and were unable to transcribe genes of the FaRLiP gene cluster. We conclude that RfpA, RfpB, and RfpC constitute a FRL-activated signal transduction cascade that is the master control switch for the FaRLiP response. FRL is proposed to activate (or inactivate) the histidine kinase activity of RfpA, which leads to formation of the active state of RfpB, the key response regulator and transcription activator. RfpC may act as a phosphate shuttle between RfpA and RfpB. Our results show that reverse genetics via conjugation will be a powerful approach in detailed studies of the FaRLiP response.

  9. Energies and spin states of FeS(0/-), FeS2(0/-), Fe2S2(0/-), Fe3S4(0/-), and Fe4S4(0/-) clusters.

    PubMed

    Li, Yan-Ni; Wang, Shengguang; Wang, Tao; Gao, Rui; Geng, Chun-Yu; Li, Yong-Wang; Wang, Jianguo; Jiao, Haijun

    2013-04-15

    The structures and energies of the electronic ground states of the FeS(0/-), FeS2(0/-), Fe2S2(0/-), Fe3S4(0/-), and Fe4S4(0/-) neutral and anionic clusters have been computed systematically with nine computational methods in combination with seven basis sets. The computed adiabatic electronic affinities (AEA) have been compared with available experimental data. Most reasonable agreements between theory and experiment have been found for both hybrid B3LYP and B3PW91 functionals in conjugation with 6-311+G* and QZVP basis sets. Detailed comparisons between the available experimental and computed AEA data at the B3LYP/6-311+G* level identified the electronic ground state of (5)Δ for FeS, (4)Δ for FeS(-), (5)B2 for FeS2, (6)A1 for FeS2(-), (1)A1 for Fe2S2, (8)A' for Fe2S2(-), (5)A'' for Fe3S4, (6)A'' for Fe3S4(-), (1)A1 for Fe4S4, and (1)A2 for Fe4S4(-). In addition, Fe2S2, Fe3S4, Fe3S4(-), Fe4S4, and Fe4S4(-) are antiferromagnetic at the B3LYP/6-311+G* level. The magnetic properties are discussed on the basis of natural bond orbital analysis. PMID:23471871

  10. Impact behavior of FeAl alloy FA-350

    SciTech Connect

    Alexander, D.J.

    1994-09-01

    The tensile properties and impact behavior of the iron aluminide FeAl-type alloy FA-350 [Fe-35.8Al-0.05Zr-0.24B (at. %)] have been studied over the temperature range of {minus}100 to 800C. Half-size Charpy specimens were either oil quenched from 700C or furnace cooled. The energy absorbed during the impact test showed a maximum value at 100 to 200C, with decreasing energy as the temperature was increased, for both heat treatments. The furnace-cooled material had greater energy absorption than the oil-quenched material. The tensile tests showed increasing ductility (as measured by total elongation) with increasing temperature. The furnace-cooled material had lower strength and higher ductility than the oil-quenched material. Fractographic examination of the oil-quenched impact specimens revealed that several different fracture modes operated, depending on the test temperature. Fracture occurred by intergranular and quasicleavage fracture at low temperatures, predominantly quasicleavage at intermediate temperatures, and intergranular fracture at 800C. For the furnace-cooled material fracture was predominantly quasicleavage at all temperatures. The higher ductility and energy absorption for the furnace-cooled material is believed to be the result of softening due to a decrease in the retained vacancy concentration.

  11. Exterior building details of Building D, east façade: painted concrete ...

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

    Exterior building details of Building D, east façade: painted concrete east face façade, main entry has flat cement plaster surround, double door six light over panels, two light transom over double door; westerly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  12. A ferulic acid (FA)-eluting system for biodegradable magnesium stent: Cells response of HUVECs.

    PubMed

    Zhang, Erlin; Shen, Feng

    2015-08-01

    A new drug-eluting system was designed for biodegradable magnesium stents, in which ferulic acid (FA) was used as drug due to its promotion function to endothelial cells and PHBHHx as drug delivery due to its biodegradability and biocompatibility. A 5 and 10% FA were added in PHBHHx to prepare FA containing PHBHHX films. The cell adhesion, spreading and proliferation on these films was investigated in order to assess cell response of HUVECs. It was also found that FA enhanced the adhesion, spreading and proliferation of HUVECs in a dose-dependent manner. Cell viability after H2 O2 injury and NO production of HUVECs were also studied. The results indicated that FA effectively inhibited H2 O2 -induced injury and promotes NO production. It was also shown that alkali treatment improved the cell adhesion, spreading and proliferation while the treatment reduces the FA release and in turn reduces the inhibition on H2 O2 -induced injury and NO production. However, alkali treatment itself had no influence on the H2 O2 induced injure and NO production. The tensile shear strength between the FA containing coating and Mg substrate was also tested. All results demonstrated that FA containing PHBHHx films exhibited strong promotion to the endothelialization and could be a choice for surface modification of magnesium stent. PMID:25630748

  13. Yacon diet (Smallanthus sonchifolius, Asteraceae) improves hepatic insulin resistance via reducing Trb3 expression in Zucker fa/fa rats

    PubMed Central

    Satoh, H; Audrey Nguyen, M T; Kudoh, A; Watanabe, T

    2013-01-01

    Objective: Yacon is a perennial plant forming a clump of >20 big, edible underground tubers. Yacon, which originates from South America, has become increasingly popular in the Japanese diet for tubers have a lower caloric value and a high fiber content. Recent studies have suggested that yacon feeding ameliorates diabetes as indicated by reduced blood glucose. Methods: We fed male Zucker fa/fa rats for 5 weeks with isocaloric normal chow diet containing from 6.5% control aroid or 6.5% yacon. Insulin sensitivity was evaluated by euglycemic-hyperinsulinemic clamp study. Results: Body weight was comparable between yacon- and aroid-fed rats. In the basal state, yacon feeding had an effect to lower fasting glucose levels from 184.1±4.1 to 167.8±2.7 mg dl−1 (P<0.01), as well as basal hepatic glucose output (HGO) from 9.9±0.4 to 7.4 ± 0.2 mg kg−1 per min (P<0.01). During the clamp studies, the glucose infusion rate required to maintain euglycemia was increased by 12.3% in yacon-fed rat. The insulin suppression of HGO was also increased in yacon-fed rats compared with control rats (85.3±2.4% vs 77.0±3.0% P<0.05), whereas the glucose disposal rate was not different between the two groups. Consistent with the clamp data, the insulin-stimulated phosphorylation of Akt was significantly enhanced in liver but not in skeletal muscle. Furthermore, tribbles 3 (Trb3) expression, which is a negative regulator of Akt activity, was markedly reduced in the liver of yacon-fed rats compared with control rats. Conclusion: These results indicate that the effect of yacon feeding to reduce blood glucose is likely due to its beneficial effects on hepatic insulin sensitivity in the insulin resistant state. PMID:23712282

  14. The MS6001FA gas turbine in mid-size combined cycle and cogeneration applications

    SciTech Connect

    Ruegger, W.A.; Anderson, R.O.

    1994-12-31

    The MS6001FA gas turbine is the latest addition to the F-technology family of gas turbines. The design is based on an aerodynamic scaling of the proven MS7001FA and MS9001FA products and is available in both 50 and 60 Hz configurations. As a result of its higher F-technology firing temperature, the 6FA is ideally suited for combined cycle and other heat recovery applications where its performance represents a significant improvement over previously available mid-size gas turbines. This paper describes the basic design of the MS6001FA, including its auxiliary systems. The gas turbine`s performance in simple cycle, combined cycle, repowering, and cogeneration applications is also reviewed.

  15. Cellular Uptake and Antitumor Activity of DOX-hyd-PEG-FA Nanoparticles

    PubMed Central

    Na, Ren; Song, Yan-feng; Mei, Qi-bing; Zhao, Ming-gao; Zhou, Si-yuan

    2014-01-01

    A PEG-based, folate mediated, active tumor targeting drug delivery system using DOX-hyd-PEG-FA nanoparticles (NPs) were prepared. DOX-hyd-PEG-FA NPs showed a significantly faster DOX release in pH 5.0 medium than in pH 7.4 medium. Compared with DOX-hyd-PEG NPs, DOX-hyd-PEG-FA NPs increased the intracellular accumulation of DOX and showed a DOX translocation from lysosomes to nucleus. The cytotoxicity of DOX-hyd-PEG-FA NPs on KB cells was much higher than that of free DOX, DOX-ami-PEG-FA NPs and DOX-hyd-PEG NPs. The cytotoxicity of DOX-hyd-PEG-FA NPs on KB cells was attenuated in the presence of exogenous folic acid. The IC50 of DOX-hyd-PEG-FA NPs and DOX-hyd-PEG NPs on A549 cells showed no significant difference. After DOX-hyd-PEG-FA NPs were intravenously administered, the amount of DOX distributed in tumor tissue was significantly increased, while the amount of DOX distributed in heart was greatly decreased as compared with free DOX. Compared with free DOX, NPs yielded improved survival rate, prolonged life span, delayed tumor growth and reduced the cardiotoxicity in tumor bearing mice model. These results indicated that the acid sensitivity, passive and active tumor targeting abilities were likely to act synergistically to enhance the drug delivery efficiency of DOX-hyd-PEG-FA NPs. Therefore, DOX-hyd-PEG-FA NPs are a promising drug delivery system for targeted cancer therapy. PMID:24828815

  16. Integrated Tail Buffet Loads on the F/A-18

    NASA Technical Reports Server (NTRS)

    Meyn, Larry A.; James, Kevin D.

    1994-01-01

    The unsteady pressures acting on the vertical tails of a full-scale F/A-18 fighter aircraft were studied to gain a better understanding of tail-buffet loads that frequently occur on fighter aircraft operating at high angles-of-attack. Data for the study were acquired during two test entries in the 80- by 120-Foot Wind Tunnel wherein the aircraft was tested at wind speeds up to 100 knots and at angles-of-attack from 20deg to 40deg. For the purposes of this paper, the primary difference between the two tests is that, during the first wind-tunnel entry, the pressure transducers were more sparsely spaced and covered less of the fin than during the second entry. In addition to a brief description of the spectral analysis methods used for the unsteady aerodynamic pressures and loads, an analysis of the effects of sensor density on estimating integrated loads is presented. It was found that the integrated loads determined from sparse sensor arrays are significantly higher than actual loads. However, a modest increase in the number of sensors can greatly reduce the error and a method for correcting load estimates from sparse sensor arrays is also suggested. The results for the time-averaged, power-spectral analysis are then presented for the tail-fin bending moments. Power spectra are presented for the aircraft at zero sideslip over an angle-of-attack range from 20deg to 40deg and for the aircraft at an angle-of-attack of 30deg over a sideslip range from -16deg to 16deg. Since the aircraft was equipped with a removable leading-edge extension (LEX) fence to reduce tail-buffet loads, the tail-fin bending moment loads are also presented for that configuration. The LEX fence is shown to significantly reduce bending moment loads over a broad range frequencies, for all the aircraft attitudes presented.

  17. Human Mitochondrial Chaperone (mtHSP70) and Cysteine Desulfurase (NFS1) Bind Preferentially to the Disordered Conformation, Whereas Co-chaperone (HSC20) Binds to the Structured Conformation of the Iron-Sulfur Cluster Scaffold Protein (ISCU)*

    PubMed Central

    Cai, Kai; Frederick, Ronnie O.; Kim, Jin Hae; Reinen, Nichole M.; Tonelli, Marco; Markley, John L.

    2013-01-01

    Human ISCU is the scaffold protein for mitochondrial iron-sulfur (Fe-S) cluster biogenesis and transfer. NMR spectra have revealed that ISCU populates two conformational states; that is, a more structured state (S) and a partially disordered state (D). We identified two single amino acid substitutions (D39V and N90A) that stabilize the S-state and two (D39A and H105A) that stabilize the D-state. We isolated the two constituent proteins of the human cysteine desulfurase complex (NFS1 and ISD11) separately and used NMR spectroscopy to investigate their interaction with ISCU. We found that ISD11 does not interact directly with ISCU. By contrast, NFS1 binds preferentially to the D-state of ISCU as does the NFS1-ISD11 complex. An in vitro Fe-S cluster assembly assay showed that [2Fe-2S] and [4Fe-4S] clusters are assembled on ISCU when catalyzed by NFS1 alone and at a higher rate when catalyzed by the NFS1-ISD11 complex. The DnaK-type chaperone (mtHSP70) and DnaJ-type co-chaperone (HSC20) are involved in the transfer of clusters bound to ISCU to acceptor proteins in an ATP-dependent reaction. We found that the ATPase activity of mtHSP70 is accelerated by HSC20 and further accelerated by HSC20 plus ISCU. NMR studies have shown that mtHSP70 binds preferentially to the D-state of ISCU and that HSC20 binds preferentially to the S-state of ISCU. PMID:23940031

  18. Identification of the iron-sulfur clusters in a ferredoxin from the archaeon Sulfolobus acidocaldarius. Evidence for a reduced [3Fe-4S] cluster with pH-dependent electronic properties.

    PubMed

    Breton, J L; Duff, J L; Butt, J N; Armstrong, F A; George, S J; Pétillot, Y; Forest, E; Schäfer, G; Thomson, A J

    1995-11-01

    A ferredoxin isolated from the archaeon Sulfolobus acidocaldarius strain DSM 639 has been shown to contain one [3Fe-4S]1 + 10 cluster with a reduction potential of -275 mV and one [4Fe-4S]2+/1+ cluster with a reduction potential of -529 mV at pH 6.4, in the temperature range 0-50 degrees C. The monomer molecular mass was confirmed to be 10907.5 +/- 1.0 Da by electrospray mass spectrometry, as calculated from the published amino acid sequence [Minami, Y. Wakabayashi. S., Wada, K., Matsubara, H., Kerscher, L. & Oesterhelt, D. (1985) J. Biochem. (Tokyo) 97, 745-751], while the holoprotein molecular mass was found to be 11,550 +/- 1.0 Da. The reduced [3Fe-4S]0 cluster was also shown by direct electrochemistry and magnetic circular dichroic spectroscopy to undergo a one-proton uptake reaction as first observed for Azotobacter chroococcum ferredoxin I [George, S. J., Richards, A. J. M., Thomson, A. J. & Yates, M. G. (1984) Biochem. J. 224, 247-251]. The pKa of the protonation step has been determined by a novel thin film electrochemical method to be 5.8. This is significantly different from the pKa of 7.7 determined for A. vinelandii ferredoxin I [Shen, B., Martin, L. L., Butt, J. N., Armstrong, F. A., Stout, C. D., Jensen, J. M., Stephens, P. J., LaMar, G. N., Gorst, C. M. & Burgess, B. K. (1993) J. Biol. Chem. 268, 25928-25939] and indicates that the polypeptide chain around the [3Fe-4S] cluster controls this reaction. Although this appears to be only the second reported case of protonation at or near the reduced [3Fe-4S]0 cluster, its observation in S. acidocaldarius ferredoxin raises the question of the generality of this chemistry for 3Fe clusters. The similarity of the pKa to the estimated intracellular pH of S. acidocaldarius strongly suggests a physiological role for this process. PMID:8521862

  19. Molecular Characterization of a Strawberry FaASR Gene in Relation to Fruit Ripening

    PubMed Central

    Jiang, Yue-ming; Zhao, Ming-lei; Shan, Wei; Kuang, Jian-fei; Lu, Wang-jin

    2011-01-01

    Background ABA-, stress- and ripening-induced (ASR) proteins have been reported to act as a downstream component involved in ABA signal transduction. Although much attention has been paid to the roles of ASR in plant development and stress responses, the mechanisms by which ABA regulate fruit ripening at the molecular level are not fully understood. In the present work, a strawberry ASR gene was isolated and characterized (FaASR), and a polyclonal antibody against FaASR protein was prepared. Furthermore, the effects of ABA, applied to two different developmental stages of strawberry, on fruit ripening and the expression of FaASR at transcriptional and translational levels were investigated. Methodology/Principal Findings FaASR, localized in the cytoplasm and nucleus, contained 193 amino acids and shared common features with other plant ASRs. It also functioned as a transcriptional activator in yeast with trans-activation activity in the N-terminus. During strawberry fruit development, endogenous ABA content, levels of FaASR mRNA and protein increased significantly at the initiation of ripening at a white (W) fruit developmental stage. More importantly, application of exogenous ABA to large green (LG) fruit and W fruit markedly increased endogenous ABA content, accelerated fruit ripening, and greatly enhanced the expression of FaASR transcripts and the accumulation of FaASR protein simultaneously. Conclusions These results indicate that FaASR may be involved in strawberry fruit ripening. The observed increase in endogenous ABA content, and enhanced FaASR expression at transcriptional and translational levels in response to ABA treatment might partially contribute to the acceleration of strawberry fruit ripening. PMID:21915355

  20. Distinct Roles of the Salmonella enterica Serovar Typhimurium CyaY and YggX Proteins in the Biosynthesis and Repair of Iron-Sulfur Clusters

    PubMed Central

    Velayudhan, Jyoti; Karlinsey, Joyce E.; Frawley, Elaine R.; Becker, Lynne A.; Nartea, Margaret

    2014-01-01

    Labile [4Fe-4S]2+ clusters found at the active sites of many dehydratases are susceptible to damage by univalent oxidants that convert the clusters to an inactive [3Fe-4S]1+ form. Bacteria repair damaged clusters in a process that does not require de novo protein synthesis or the Isc and Suf cluster assembly pathways. The current study investigates the participation of the bacterial frataxin ortholog CyaY and the YggX protein, which are proposed to play roles in iron trafficking and iron-sulfur cluster repair. Previous reports found that individual mutations in cyaY or yggX were not associated with phenotypic changes in Escherichia coli and Salmonella enterica serovar Typhimurium, suggesting that CyaY and YggX might have functionally redundant roles. However, we have found that individual mutations in cyaY or yggX confer enhanced susceptibility to hydrogen peroxide in Salmonella enterica serovar Typhimurium. In addition, inactivation of the stm3944 open reading frame, which is located immediately upstream of cyaY and which encodes a putative inner membrane protein, dramatically enhances the hydrogen peroxide sensitivity of a cyaY mutant. Overexpression of STM3944 reduces the elevated intracellular free iron levels observed in an S. Typhimurium fur mutant and also reduces the total cellular iron content under conditions of iron overload, suggesting that the stm3944-encoded protein may mediate iron efflux. Mutations in cyaY and yggX have different effects on the activities of the iron-sulfur cluster-containing aconitase, serine deaminase, and NADH dehydrogenase I enzymes of S. Typhimurium under basal conditions or following recovery from oxidative stress. In addition, cyaY and yggX mutations have additive effects on 6-phosphogluconate dehydratase-dependent growth during nitrosative stress, and a cyaY mutation reduces Salmonella virulence in mice. Collectively, these results indicate that CyaY and YggX play distinct supporting roles in iron-sulfur cluster biosynthesis

  1. FaRe: A Mathematica package for tensor reduction of Feynman integrals

    NASA Astrophysics Data System (ADS)

    Re Fiorentin, Michele

    2016-08-01

    In this paper, we present FaRe, a package for Mathematica that implements the decomposition of a generic tensor Feynman integral, with arbitrary loop number, into scalar integrals in higher dimension. In order for FaRe to work, the package FeynCalc is needed, so that the tensor structure of the different contributions is preserved and the obtained scalar integrals are grouped accordingly. FaRe can prove particularly useful when it is preferable to handle Feynman integrals with free Lorentz indices and tensor reduction of high-order integrals is needed. This can then be achieved with several powerful existing tools.

  2. 3. Historic American Buildings Survey, George F.A. Palmer, Photographer, June ...

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

    3. Historic American Buildings Survey, George F.A. Palmer, Photographer, June 16, 1936 DETAIL OF HOPPER AND HOISTING MECHANISM (FOR REMOVAL OF MILL STONES. 1757, CUT IN STONE). - Benjamin Hammond Grist Mill, Hammond Hill Road, Kingston, Washington County, RI

  3. 4. Historic American Buildings Survey, George F.A. Palmer, Photographer, June ...

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

    4. Historic American Buildings Survey, George F.A. Palmer, Photographer, June 16, 1936 DETAIL OF RECEIVING TROUGH (BASEMENT). - Benjamin Hammond Grist Mill, Hammond Hill Road, Kingston, Washington County, RI

  4. Exterior building details of Building D, west façade: brick arch ...

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

    Exterior building details of Building D, west façade: brick arch lintel over historic entry that was brick infilled; easterly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  5. Exterior building details of Building D, west façade: doublehung wood ...

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

    Exterior building details of Building D, west façade: double-hung wood window with brick arch lintel and brick sill; easterly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  6. Exterior building details of Building E, oblique west façade: brick ...

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

    Exterior building details of Building E, oblique west façade: brick arch lintel and brick infilled window with brick sill; southeasterly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  7. North façade of crucible steel building; looking southwest Bethlehem ...

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

    North façade of crucible steel building; looking southwest - Bethlehem Steel Corporation, South Bethlehem Works, Crucible Steel Plant, Along Lehigh River, North of Fourth Street, West of Minsi Trail Bridge, Bethlehem, Northampton County, PA

  8. Performance evaluation of a low-temperature solar Rankine cycle system utilizing R245fa

    SciTech Connect

    Wang, X.D.; Zhao, L.; Wang, J.L.; Zhang, W.Z.; Zhao, X.Z.; Wu, W.

    2010-03-15

    A low-temperature solar Rankine system utilizing R245fa as the working fluid is proposed and an experimental system is designed, constructed and tested. Both the evacuated solar collectors and the flat plate solar collectors are used in the experimental system; meanwhile, a rolling-piston R245fa expander is also mounted in the system. The new designed R245fa expander works stably in the experiment, with an average expansion power output of 1.73 kW and an average isentropic efficiency of 45.2%. The overall power generation efficiency estimated is 4.2%, when the evacuated solar collector is utilized in the system, and with the condition of flat plate solar collector, it is about 3.2%. The experimental results show that using R245fa as working fluid in the low-temperature solar power Rankine cycle system is feasible and the performance is acceptable. (author)

  9. Georgia Power Co. Quantities for F.A. P. 425G (Bridge) Contract ...

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

    Georgia Power Co. Quantities for F.A. P. 425G (Bridge) Contract No. 1 and 2 - Tallulah Falls Bridge, Spanning Tallulah Falls River on U.S. Highway 23/State Route 15, Tallulah Falls, Habersham County, GA

  10. West façade of shop building, looking east Bethlehem Steel ...

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

    West façade of shop building, looking east - Bethlehem Steel Corporation, South Bethlehem Works, Tool Steel-Electric Furnace Shop, Along Lehigh River, North of Fourth Street, West of Minsi Trail Bridge, Bethlehem, Northampton County, PA

  11. Exterior building details of Building B, east façade: second floor ...

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

    Exterior building details of Building B, east façade: second floor entrance with cement plaster profiled surround and embedded wood beam end; westerly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  12. NASA F/A-18 Flight-Tests Mars Science Lab Radar

    NASA Video Gallery

    A NASA F/A-18 put the Jet Propulsion Laboratory's Mars Science Laboratory landing radar to the test during recent flights over Edwards Air Force Base, Calif. With the test radar housed in a pod und...

  13. View north detail of south façade showing damage to wall ...

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

    View north detail of south façade showing damage to wall - Naval Base Philadelphia-Philadelphia Naval Shipyard, Foundry-Propeller Shop, North of Porter Avenue, west of Third Street West, Philadelphia, Philadelphia County, PA

  14. View northeast; detail of southwest corner showing damage to façade ...

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

    View northeast; detail of southwest corner showing damage to façade - Naval Base Philadelphia-Philadelphia Naval Shipyard, Foundry-Propeller Shop, North of Porter Avenue, west of Third Street West, Philadelphia, Philadelphia County, PA

  15. Exterior building details of Building A; north façade: iron latticed ...

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

    Exterior building details of Building A; north façade: iron latticed gate dungeon entrance, granite base; southerly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  16. MISCIBILITY, SOLUBILITY, VISCOSITY, AND DENSITY MEASUREMENTS FOR R-236FA WITH POTENTIAL LUBRICANTS.

    EPA Science Inventory

    Miscibility, solubility, viscosity, and density data are needed to determine the suitability of refrigerant/lubricant combinations for use in refrigeration systems. These property data have been obtained for HFC (hydrofluorocarbon) -236fa and two potential lubricants. The tested ...

  17. NASA Videographer Captures X-51 from F/A-18 Chase

    NASA Video Gallery

    A tiny dashcam video camera records NASA Dryden videographer Lori Losey capturing the historic launch of the Air Force's X-51 Waverider hypersonic vehicle from the rear cockpit of a NASA F/A-18 flo...

  18. East façade, Burton Park Club House, with Amphitheater in foreground, ...

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

    East façade, Burton Park Club House, with Amphitheater in foreground, view to north from Amphitheater stage (90 mm lens). - Burton Park, Club House & Amphitheater, Adjacent ot south end of Chestnut Avenue, San Carlos, San Mateo County, CA

  19. Nearly monotonic problems: A key to effective FA/C distributed sensor interpretation?

    SciTech Connect

    Carver, N.; Lesser, V.; Whitehair, R.

    1996-12-31

    The functionally-accurate, cooperative (FA/C) distributed problem-solving paradigm is one approach for organizing distributed problem solving among homogeneous, cooperating agents. A key assumption of the FA/C model has been that the agents` local solutions can substitute for the raw data in determining the global solutions. This is not the case in general, however. Does this mean that researchers` intuitions have been wrong and/or that FA/C problem solving is not likely to be effective? We suggest that some domains have a characteristic that can account for the success of exchanging mainly local solutions. We call such problems nearly monotonic. This concept is discussed in the context of FA/C-based distributed sensor interpretation.

  20. Protein labeling with the labeling precursor [(18)F]SiFA-SH for positron emission tomography.

    PubMed

    Wängler, Björn; Kostikov, Alexey P; Niedermoser, Sabrina; Chin, Joshua; Orchowski, Katy; Schirrmacher, Esther; Iovkova-Berends, Liuba; Jurkschat, Klaus; Wängler, Carmen; Schirrmacher, Ralf

    2012-11-01

    Proteins previously derivatized with the cross-coupling reagent sulfo-SMCC (4-(N-maleimidomethyl)cyclohexane-1-carboxylic acid 3-sulfo-N-hydroxy-succinimide ester sodium salt) can be easily labeled in high radiochemical yields with the silicon-fluoride acceptor (SiFA) reagent [(18)F]SiFA-SH, obtained via isotopic exchange, by thiol-maleimide coupling chemistry (n = 10). The specific activity of SiFA-SH obtained in a one-step labeling reaction was > 18.5 GBq μmol(-1) (> 500 Ci mmol(-1)). The number of SiFA building blocks per protein molecule is defined by the previously introduced number of maleimide groups, which can be determined by a simple and convenient Ellman's assay. Not more than two maleimide groups are introduced using sulfo-SMCC, thereby keeping the modification of the protein low and preserving its biological activity. PMID:23037311

  1. East and north elevations (rear façade) of quarters no. 2, ...

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

    East and north elevations (rear façade) of quarters no. 2, looking southwest. - Sacramento National Wildlife Refuge, Headquarters Complex, Quarters No. 2, 752 County Road 99W, Willows, Glenn County, CA

  2. Exterior building details of Building A; north façade: fouroverfour doublehung ...

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

    Exterior building details of Building A; north façade: four-over-four double-hung wood sash window with concrete sill; southerly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  3. Exterior building details of Building D, west façade: second floor ...

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

    Exterior building details of Building D, west façade: second floor metal multi-pane industrial-type sash windows; easterly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  4. View of W.I. Elliott Building, 16th Street façade, detail of ...

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

    View of W.I. Elliott Building, 16th Street façade, detail of typical bay with storefront, view from 16th Street, facing west. - W.I. Elliott Building, 1530 J Street, Sacramento, Sacramento County, CA

  5. CLUSTER CHEMISTRY

    SciTech Connect

    Muetterties, Earl L.

    1980-05-01

    Metal cluster chemistry is one of the most rapidly developing areas of inorganic and organometallic chemistry. Prior to 1960 only a few metal clusters were well characterized. However, shortly after the early development of boron cluster chemistry, the field of metal cluster chemistry began to grow at a very rapid rate and a structural and a qualitative theoretical understanding of clusters came quickly. Analyzed here is the chemistry and the general significance of clusters with particular emphasis on the cluster research within my group. The importance of coordinately unsaturated, very reactive metal clusters is the major subject of discussion.

  6. Exterior building details of Building B, east façade: ellshaped south ...

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

    Exterior building details of Building B, east façade: ell-shaped south facing concrete staircase with decorative pipe railing, second floor entrance with cement plaster profiled surround, dentil course cornice, truncated embedded wood beams, cream colored plaster-finished exterior façade, closed off window well with protruding vent; northwesterly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  7. Clustering structure of nuclei in deep inelastic processes

    NASA Astrophysics Data System (ADS)

    Hirai, M.; Kumano, S.; Saito, K.; Watanabe, T.

    2014-12-01

    A clustering aspect is explained for the 9Be nucleus in charged-lepton deep inelastic scattering. Nuclear modifications of the structure function F2 are studied by the ratio REMC = FA2/FD2, where A and D are a nucleus and the deuteron, respectively. In a JLab experiment, an unexpectedly large nuclear modification slope |dREMC/dx| was found for 9 Be, which could be related to its clustering structure. We investigated a mean conventional part of a nuclear structure function FA2 by a convolution description with nucleon momentum distributions calculated by antisymmetrized (or fermionic) molecular dynamics (AMD) and also by a simple shell model. We found that clustering effects are small in the conventional part, so that the JLab result could be associated with an internal nucleon modification or a short-range nuclear correlation which is caused by high densities due to cluster formation.

  8. Variability of Southern Valu Fa Ridge Magmatic Systems

    NASA Astrophysics Data System (ADS)

    Goddard, C. I.; Christie, D. M.; Arculus, R.

    2004-12-01

    Valu Fa Ridge (VFR), which encompasses the southernmost segments of the East Lau Spreading Center (ELSC), is an important end-member in the spectrum of back-arc spreading centers because it is strongly affected in all aspects of the spreading process by inputs from the nearby active Tofua (Tonga) volcanic arc, and because its magma systems are rapidly evolving as the VFR propagates to the south. New lava samples collected by the TELVE Expedition of the R/V Southern Surveyor (Australia) from the four southernmost VFR segments have greatly increased the number and spatial distribution of fresh volcanic glass samples, quadrupling the availability of "primitive" (MgO > 6 wt.%) glass samples and encompassing significant along- and across-axis geochemical variability. These new data provide an opportunity to evaluate both the evolution of crustal magmatic processes relative to southward rift propagation and variability in mantle source inputs relative to the active volcanic arc. The four sampled segments of VFR are separated by left-stepping overlapping offsets that differ from their mid-ocean ridge counterparts in their longer, more parallel overlapping limbs and in the absence of an overlap basin. The TELVE glasses display considerable major element variability and the VFR is unusual among well-developed spreading centers in its strongly bimodal volcanism, in the abundance of evolved lavas and in the coexistence of two distinct liquid lines of descent. Dacites and rhyolites (SiO2 ~67-75 wt.%) are relatively abundant close to segment ends or discontinuities along the southernmost three ridge segments, but rare from the northernmost sampled segment and from off-axis seamounts. A "Daly Gap" from ~60-67 wt.% SiO2 and 1.75-0.75 wt.% MgO is present along the northern segments but absent near the southern propagating rift tip. Primitive (MgO > 6wt.%) glasses were recovered from all VFR segments and on 5 of the 9 sampled seamounts. Both high-silica glasses and FeTi basalt

  9. Architectural Kansei of ‘Wall’ in The Façade Design by Le Corbusier

    NASA Astrophysics Data System (ADS)

    Sendai, Shoichiro

    The purpose of this paper is to discuss the modern architect Le Corbusier's architectural Kansei (sensibility) on wall in site environment through the analysis of his façade design, using Œuvres complètes (1910-1965, 8 vols., Les éditions d'architecture, Artemis, Zurich) and Le Corbusier Archives (1982-1984, 32 vols., Garland Publishing, Inc. and Fondation Le Corbusier, New York, London, Paris). At first, I arrange five façade types, according to the explanation by Le Corbusier ; ‘fenêtre en longueur (strip window)’, ‘pan de verre (glass wall)’, ‘brise-soleil (sun-breaker)’, ‘loggia’ and ‘claustra’. Through the analysis of the relationship between these types and the design process of each building, we find that Le Corbusier's façade design includes the affirmation and the negation of the ‘wall’ at the same time. In fact, the nature of façade modification during design process is divers: increase in transparency, decrease in transparency and spatialization of façade. That means, Le Corbusier studied the environmental condition by these façade types, and tried to realize the phenomenal openness. This trial bases on the function of architectural Kansei as correspondence between body and environment beyond the physical design.

  10. FANCI Regulates Recruitment of the FA Core Complex at Sites of DNA Damage Independently of FANCD2

    PubMed Central

    Castella, Maria; Jacquemont, Celine; Thompson, Elizabeth L.; Yeo, Jung Eun; Cheung, Ronald S.; Huang, Jen-Wei; Sobeck, Alexandra; Hendrickson, Eric A.; Taniguchi, Toshiyasu

    2015-01-01

    The Fanconi anemia (FA)-BRCA pathway mediates repair of DNA interstrand crosslinks. The FA core complex, a multi-subunit ubiquitin ligase, participates in the detection of DNA lesions and monoubiquitinates two downstream FA proteins, FANCD2 and FANCI (or the ID complex). However, the regulation of the FA core complex itself is poorly understood. Here we show that the FA core complex proteins are recruited to sites of DNA damage and form nuclear foci in S and G2 phases of the cell cycle. ATR kinase activity, an intact FA core complex and FANCM-FAAP24 were crucial for this recruitment. Surprisingly, FANCI, but not its partner FANCD2, was needed for efficient FA core complex foci formation. Monoubiquitination or ATR-dependent phosphorylation of FANCI were not required for the FA core complex recruitment, but FANCI deubiquitination by USP1 was. Additionally, BRCA1 was required for efficient FA core complex foci formation. These findings indicate that FANCI functions upstream of FA core complex recruitment independently of FANCD2, and alter the current view of the FA-BRCA pathway. PMID:26430909

  11. The cathepsin B inhibitor, z-FA-CMK is toxic and readily induced cell death in human T lymphocytes

    SciTech Connect

    Liow, K.Y.; Chow, S.C.

    2013-11-01

    The cathepsin B inhibitor, benzyloxycarbonyl-phenylalanine-alanine-chloromethylketone (z-FA-CMK) was found to be toxic and readily induced cell death in the human T cell line, Jurkat, whereas two other analogs benzyloxycarbonyl-phenylalanine-alanine-fluoromethylketone (z-FA-FMK) and benzyloxycarbonyl-phenylalanine-alanine-diazomethylketone (z-FA-DMK) were not toxic. The toxicity of z-FA-CMK requires not only the CMK group, but also the presence of alanine in the P1 position and the benzyloxycarbonyl group at the N-terminal. Dose–response studies showed that lower concentrations of z-FA-CMK induced apoptosis in Jurkat T cells whereas higher concentrations induced necrosis. In z-FA-CMK-induced apoptosis, both initiator caspases (-8 and -9) and effector caspases (-3, -6 and -7) were processed to their respective subunits in Jurkat T cells. However, only the pro-form of the initiator caspases were reduced in z-FA-CMK-induced necrosis and no respective subunits were apparent. The caspase inihibitor benzyloxycarbonyl-valine-alanine-aspartic acid-(O-methyl)-fluoromehylketone (z-VAD-FMK) inhibits apoptosis and caspase processing in Jurkat T cells treated with low concentration of z-FA-CMK but has no effect on z-FA-CMK-induced necrosis and the loss of initiator caspases. This suggests that the loss of initiator caspases in Jurkat T cells during z-FA-CMK-induced necrosis is not a caspase-dependent process. Taken together, we have demonstrated that z-FA-CMK is toxic to Jurkat T cells and induces apoptosis at low concentrations, while at higher concentrations the cells die of necrosis. - Highlights: • z-FA-CMK is toxic and induce cell death in the human T cells. • z-FA-CMK toxicity requires the CMK group, alanine and the benzyloxycarbonyl group. • z-FA-CMK induced apoptosis at low concentration and necrosis at high concentration.

  12. Berberine-induced anticancer activities in FaDu head and neck squamous cell carcinoma cells.

    PubMed

    Seo, Yo-Seob; Yim, Min-Ji; Kim, Bok-Hee; Kang, Kyung-Rok; Lee, Sook-Young; Oh, Ji-Su; You, Jae-Seek; Kim, Su-Gwan; Yu, Sang-Joun; Lee, Gyeong-Je; Kim, Do Kyung; Kim, Chun Sung; Kim, Jin-Soo; Kim, Jae-Sung

    2015-12-01

    In the present study, we investigated berberine‑induced apoptosis and the signaling pathways underlying its activity in FaDu head and neck squamous cell carcinoma cells. Berberine did not affect the viability of primary human normal oral keratinocytes. In contrast, the cytotoxicity of berberine was significantly increased in FaDu cells stimulated with berberine for 24 h. Furthermore, berberine increased nuclear condensation and apoptosis rates in FaDu cells than those in untreated control cells. Berberine also induced the upregulation of apoptotic ligands, such as FasL and TNF-related apoptosis-inducing ligand, and triggered the activation of caspase-8, -7 and -3, and poly(ADP ribose) polymerase, characteristic of death receptor-dependent extrinsic apoptosis. Moreover, berberine activated the mitochondria‑dependent apoptotic signaling pathway by upregulating pro-apoptotic factors, such as Bax, Bad, Apaf-1, and the active form of caspase-9, and downregulating anti-apoptotic factors, such as Bcl-2 and Bcl-xL. In addition, berberine increased the expression of the tumor suppressor p53 in FaDu cells. The pan-caspase inhibitor Z-VAD-fmk suppressed the activation of caspase-3 and prevented cytotoxicity in FaDu cells treated with berberine. Interestingly, berberine suppressed cell migration through downregulation of vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-2, and MMP-9. Moreover, the phosphorylation of extracellular signal-regulated kinase (ERK1/2) and p38, components of the mitogen-activated protein kinase pathway that are associated with the expression of MMP and VEGF, was suppressed in FaDu cells treated with berberine for 24 h. Therefore, these data suggested that berberine exerted anticancer effects in FaDu cells through induction of apoptosis and suppression of migration. Berberine may have potential applications as a chemotherapeutic agent for the management of head and neck squamous carcinoma. PMID:26503508

  13. Novel bioactive Co-based alloy/FA nanocomposite for dental applications

    PubMed Central

    Fathi, Mohammadhossein; Ahmadian, Mehdi; Bahrami, Mojgan

    2012-01-01

    Background: Dental cobalt base alloys are biocompatible dental materials and have been widely used in dentistry. However, metals are bioinert and may not present bioactivity in human body. Bioactivity is the especial ability to interact with human body and make a bonding to soft and hard tissues. The aim of the present research was fabrication and bioactivity evaluation of novel cobalt alloy/Fluorapatite nanocomposite (CoA/FaNC) with different amounts of Fluorapatite (FA) nanopowder. Materials and Methods: Co-Cr-Mo alloy (ASTM F75) powder was prepared and mixed in a planetary ball mill with different amounts of FA nanopowders (10, 15, 20% wt). Prepared composite powders were cold pressed and sintered at 1100°C for 4 h. X-ray diffraction (XRD), scanning electron microscopy and transition electron microscopy techniques were used for phase analysis, crystallite size determination of FA and also for phase analysis and evaluation of particle distribution of composites. Bioactivity behavior of prepared nanocomposites was evaluated in simulated body fluid (SBF) for 1 up to 28 days. Materials and Methods: Co-Cr-Mo alloy (ASTM F75) powder was prepared and mixed in a planetary ball mill with different amounts of FA nanopowders (10, 15, 20% wt). Prepared composite powders were cold pressed and sintered at 1100°C for 4 h. X-ray diffraction (XRD), scanning electron microscopy and transition electron microscopy techniques were used for phase analysis, crystallite size determination of FA and also for phase analysis and evaluation of particle distribution of composites. Bioactivity behavior of prepared nanocomposites was evaluated in simulated body fluid (SBF) for 1 up to 28 days. Results: Results showed that nucleus of apatite were formed on the surface of the prepared CoA/FaNC during 1 up to 28 days immersion in the SBF solution. On the other hand, CoA/FaNC unlike Co-base alloy possessed bone-like apatite-formation ability. Conclusion: It was concluded that bioinert Co

  14. Both FA- and mPEG-conjugated chitosan nanoparticles for targeted cellular uptake and enhanced tumor tissue distribution

    PubMed Central

    2011-01-01

    Both folic acid (FA)- and methoxypoly(ethylene glycol) (mPEG)-conjugated chitosan nanoparticles (NPs) had been designed for targeted and prolong anticancer drug delivery system. The chitosan NPs were prepared with combination of ionic gelation and chemical cross-linking method, followed by conjugation with both FA and mPEG, respectively. FA-mPEG-NPs were compared with either NPs or mPEG-/FA-NPs in terms of their size, targeting cellular efficiency and tumor tissue distribution. The specificity of the mPEG-FA-NPs targeting cancerous cells was demonstrated by comparative intracellular uptake of NPs and mPEG-/FA-NPs by human adenocarcinoma HeLa cells. Mitomycin C (MMC), as a model drug, was loaded to the mPEG-FA-NPs. Results show that the chitosan NPs presented a narrow-size distribution with an average diameter about 200 nm regardless of the type of functional group. In addition, MMC was easily loaded to the mPEG-FA-NPs with drug-loading content of 9.1%, and the drug releases were biphasic with an initial burst release, followed by a subsequent slower release. Laser confocal scanning imaging proved that both mPEG-FA-NPs and FA-NPs could greatly enhance uptake by HeLa cells. In vivo animal experiments, using a nude mice xenograft model, demonstrated that an increased amount of mPEG-FA-NPs or FA-NPs were accumulated in the tumor tissue relative to the mPEG-NPs or NPs alone. These results suggest that both FA- and mPEG-conjugated chitosan NPs are potentially prolonged drug delivery system for tumor cell-selective targeting treatments. PMID:22027239

  15. Analysis and Assessment of Strength Development in Compressed FaL-G Blocks

    NASA Astrophysics Data System (ADS)

    Nagendra Prasad, K.; Vijaya Bhaskar, S.; Narasimhulu, M. L.; Manohara Reddy, R.

    2014-09-01

    Of the several options explored in large scale utilization of fly ash, such as production of blended cements, high volume fly ash cement concretes, fly ash, lime and gypsum (FaL-G) combinations, alkali activated fly ash mortars and concretes are of recent innovations. The last two are non-traditional cementing materials, since no cement is used in processing of these materials. This investigation deals with analysis and assessment of strength development in compressed FaL-G blocks. FaL-G chemistry provides a strong scientific base for understanding the mechanisms of interaction. But an equally strong technological base in the production of FaL-G blocks is the need of the hour. In this investigation, analysis has been made to advance a phenomenological model to arrive at the combinations of the ingredients to produce compressed blocks to meet the strength development desired at specified age, based on carefully planned experimental data generated. The analysis of test results has been done within the framework of Abrams' law, which is extensively used in concrete technology. The validity has been examined with an independent set of experimental data. With incorporation of more data covering still wider spectrum of materials the phenomenological model can further be reinforced as a viable tool in the production of compressed FaL-G blocks.

  16. FA-loaded lipid drug delivery systems: preparation, characterization and biological studies.

    PubMed

    Carbone, Claudia; Campisi, Agata; Musumeci, Teresa; Raciti, Giuseppina; Bonfanti, Roberta; Puglisi, Giovanni

    2014-02-14

    The main purpose of this research was to prepare and to characterize ferulic acid-loaded nanostructured lipid carrier (FA-NLC) to evaluate the cytotoxic effect on human glioblastoma cancer U87MG cells. First of all, the influence of different materials on mean size and homogeneity of NLC prepared by a low energy organic solvent-free method was investigated. Technological characterization (encapsulation efficiency, mean particle size, homogeneity and in vitro release profile) was performed on the selected NLC in comparison to others lipid carriers, nanoemulsion and SLN. Furthermore, the thermal behavior of NLC and SLN was investigated using Differential Scanning Calorimetry (DSC) in order to evaluate their structure. Biological studies (MTT bioassay and caspase-3 cleavage) on the selected NLC showed no cytotoxic effects of the unloaded tested NLC. Besides, the effectiveness of FA-loaded NLC was higher compared to the free drug. Cells treated with FA or FA-loaded NLC showed a greater effect compared to idebenone (IDE) or IDE-loaded NLC, respectively. These results strongly support that FA-loaded NLC could be potentially used for the treatment of glioblastoma. PMID:24514450

  17. Aerodynamics of powered missile separation from F/A-18 aircraft

    NASA Technical Reports Server (NTRS)

    Ahmad, J. U.; Shanks, S. P.; Buning, P. G.

    1993-01-01

    A 3D dynamic 'chimera' algorithm that solves the thin-layer Navier-Stokes equations over multiple moving bodies was modified to numerically simulate the aerodynamics, missile dynamics, and missile plume interactions of a missile separating from a generic wing and from an F/A-18 aircraft in transonic flow. The missile is mounted below the wing for missile separation from the wing and on the F/A-18 fuselage at the engine inlet side for missile separation from aircraft. Static and powered missile separation cases are considered to examine the influence of the missile and plume on the wing and F/A-18 fuselage and engine inlet. The aircraft and missile are at two degrees angle of attack, Reynolds number of 10 million, freestream Mach number of 1.05 and plume Mach number of 3.0. The computational results show the details of the flow field.

  18. Modeling the binding of fulvic acid by goethite: the speciation of adsorbed FA molecules

    NASA Astrophysics Data System (ADS)

    Filius, Jeroen D.; Meeussen, Johannes C. L.; Lumsdon, David G.; Hiemstra, Tjisse; van Riemsdijk, Willem H.

    2003-04-01

    Under natural conditions, the adsorption of ions at the solid-water interface may be strongly influenced by the adsorption of organic matter. In this paper, we describe the adsorption of fulvic acid (FA) by metal(hydr)oxide surfaces with a heterogeneous surface complexation model, the ligand and charge distribution (LCD) model. The model is a self-consistent combination of the nonideal competitive adsorption (NICA) equation and the CD-MUSIC model. The LCD model can describe simultaneously the concentration, pH, and salt dependency of the adsorption with a minimum of only three adjustable parameters. Furthermore, the model predicts the coadsorption of protons accurately for an extended range of conditions. Surface speciation calculations show that almost all hydroxyl groups of the adsorbed FA molecules are involved in outer sphere complexation reactions. The carboxylic groups of the adsorbed FA molecule form inner and outer sphere complexes. Furthermore, part of the carboxylate groups remain noncoordinated and deprotonated.

  19. Flight Tests of a Ministick Controller in an F/A-18 Airplane

    NASA Technical Reports Server (NTRS)

    Stoliker, Patrick C.; Carter, John

    2003-01-01

    In March of 1999, five pilots performed flight tests to evaluate the handling qualities of an F/A-18 research airplane equipped with a small-displacement center stick (ministick) controller that had been developed for the JAS 39 Gripen airplane (a fighter/attack/ reconnaissance airplane used by the Swedish air force). For these tests, the ministick was installed in the aft cockpit (see figure) and production support flight control computers (PSFCCs) were used as interfaces between the controller hardware and the standard F/A-18 flight-control laws. The primary objective of the flight tests was to assess any changes in handling qualities of the F/A-18 airplane attributable to the mechanical characteristics of the ministick. The secondary objective was to demonstrate the capability of the PSFCCs to support flight-test experiments.

  20. Explication of Definitional Description and Empirical Use of Fraction of Orally Administered Drugs Absorbed From the Intestine (Fa) and Intestinal Availability (Fg): Effect of P-glycoprotein and CYP3A on Fa and Fg.

    PubMed

    Tanaka, Yuta; Kitamura, Yoshiaki; Maeda, Kazuya; Sugiyama, Yuichi

    2016-02-01

    Conventionally, it is believed that the fraction of orally administered drugs absorbed from the intestine (Fa) and intestinal availability (Fg) are independently determined by the apical membrane permeation and intestinal metabolism, respectively. However, the validity of this belief has not been well discussed, and Fa and Fg are often used without careful definition. In this review, Fa and Fg are mathematically described based on their definitions under the linear kinetics of metabolism and transport. Even considering with different models, intestinal metabolic enzymes such as cytochrome P450 3A affected both Fa and Fg, whereas apical efflux transporters including P-glycoprotein had no influence on Fg at least under the linear condition. To determine whether Fa and Fg calculated using different clinical methods are identical, empirical Fa and Fg were mathematically described based on "feces method" and "grapefruit juice method" and compared with their definitions. Fa and Fg obtained by the feces method corresponded with their definitions whereas the grapefruit juice method provided smaller Fa and larger Fg particularly for dual substrates of P-glycoprotein and cytochrome P450 3A with low membrane permeability. Our analyses suggest that the definitions and calculation methods of Fa and Fg should be considered when we intend to separately determine these values. PMID:26869410

  1. Birth order and avuncular tendencies in Samoan men and fa'afafine.

    PubMed

    Vanderlaan, Doug P; Vasey, Paul L

    2013-04-01

    Androphilia refers to sexual attraction and arousal to males whereas gynephilia refers to sexual attraction and arousal to females. In Samoa, transgendered androphilic males are known locally as fa'afafine. Previous research has shown that, compared to Samoan gynephilic men, fa'afafine report greater willingness to invest time and money toward nieces and nephews (i.e., greater avuncular tendencies) and also have greater numbers of older brothers and older sisters. The present study examined whether the Samoan male sexual orientation difference in avuncular tendencies could be accounted for by these parallel differences in numbers of older brothers and older sisters. The sample included 204 fa'afafine and 272 Samoan gynephilic men from our Samoan data archive for whom we had concurrent information on (1) a measure of willingness to invest time and money in nieces and nephews (i.e., avuncular tendencies) and (2) numbers of older and younger biological brothers and sisters. Among fa'afafine, but not Samoan gynephilic men, number of older brothers and number of older sisters were both significantly positively associated with avuncular tendencies. When controlling for number of older brothers, the magnitude of the male sexual orientation difference in avuncular tendencies was lowered, but remained statistically significant. In contrast, when controlling for number of older sisters, the male sexual orientation difference in avuncular tendencies ceased to exist. Discussion detailed how these findings help hone in on the proximate basis of elevated avuncular tendencies among fa'afafine. In addition, discussion focused on how particular evolutionary and cultural factors might relate to the avuncularity of fa'afafine. PMID:23242468

  2. Detecting blind building façades from highly overlapping wide angle aerial imagery

    NASA Astrophysics Data System (ADS)

    Burochin, Jean-Pascal; Vallet, Bruno; Brédif, Mathieu; Mallet, Clément; Brosset, Thomas; Paparoditis, Nicolas

    2014-10-01

    This paper deals with the identification of blind building façades, i.e. façades which have no openings, in wide angle aerial images with a decimeter pixel size, acquired by nadir looking cameras. This blindness characterization is in general crucial for real estate estimation and has, at least in France, a particular importance on the evaluation of legal permission of constructing on a parcel due to local urban planning schemes. We assume that we have at our disposal an aerial survey with a relatively high stereo overlap along-track and across-track and a 3D city model of LoD 1, that can have been generated with the input images. The 3D model is textured with the aerial imagery by taking into account the 3D occlusions and by selecting for each façade the best available resolution texture seeing the whole façade. We then parse all 3D façades textures by looking for evidence of openings (windows or doors). This evidence is characterized by a comprehensive set of basic radiometric and geometrical features. The blindness prognostic is then elaborated through an (SVM) supervised classification. Despite the relatively low resolution of the images, we reach a classification accuracy of around 85% on decimeter resolution imagery with 60 × 40 % stereo overlap. On the one hand, we show that the results are very sensitive to the texturing resampling process and to vegetation presence on façade textures. On the other hand, the most relevant features for our classification framework are related to texture uniformity and horizontal aspect and to the maximal contrast of the opening detections. We conclude that standard aerial imagery used to build 3D city models can also be exploited to some extent and at no additional cost for facade blindness characterisation.

  3. A Security-façade Library for Virtual-observatory Software

    NASA Astrophysics Data System (ADS)

    Rixon, G.

    2009-09-01

    The security-façade library implements, for Java, IVOA's security standards. It supports the authentication mechanisms for SOAP and REST web-services, the sign-on mechanisms (with MyProxy, AstroGrid Accounts protocol or local credential-caches), the delegation protocol, and RFC3820-enabled HTTPS for Apache Tomcat. Using the façade, a developer who is not a security specialist can easily add access control to a virtual-observatory service and call secured services from an application. The library has been an internal part of AstroGrid software for some time and it is now offered for use by other developers.

  4. Doppler Global Velocimetry Measurements of the Vortical Flow Above an F/A-18

    NASA Technical Reports Server (NTRS)

    Lee, Joseph W.; Meyers, James F.; Cavone, Angelo A.; Suzuki, Karen E.

    1993-01-01

    A Doppler global velocimeter was used to investigate the vortical flow above an F/A-18 model at 25-degrees angle of attack. The measurements indicate that the flow had the same characteristics as the vortical flow above a standard delta wing. The flow pattern indicating transition from stable to burst conditions found above the delta wing was also found at the 440 station above the F/A-18. Measurements downstream at the 524 station found that the flow velocity varied considerably, with standard deviations reaching 30 percent of free stream. However, individual data images indicated that the flow was spatially coherent, and not chaotic as expected.

  5. Meaningful Clusters

    SciTech Connect

    Sanfilippo, Antonio P.; Calapristi, Augustin J.; Crow, Vernon L.; Hetzler, Elizabeth G.; Turner, Alan E.

    2004-05-26

    We present an approach to the disambiguation of cluster labels that capitalizes on the notion of semantic similarity to assign WordNet senses to cluster labels. The approach provides interesting insights on how document clustering can provide the basis for developing a novel approach to word sense disambiguation.

  6. Principles of sustained enzymatic hydrogen oxidation in the presence of oxygen--the crucial influence of high potential Fe-S clusters in the electron relay of [NiFe]-hydrogenases.

    PubMed

    Evans, Rhiannon M; Parkin, Alison; Roessler, Maxie M; Murphy, Bonnie J; Adamson, Hope; Lukey, Michael J; Sargent, Frank; Volbeda, Anne; Fontecilla-Camps, Juan C; Armstrong, Fraser A

    2013-02-20

    "Hyd-1", produced by Escherichia coli , exemplifies a special class of [NiFe]-hydrogenase that can sustain high catalytic H(2) oxidation activity in the presence of O(2)-an intruder that normally incapacitates the sulfur- and electron-rich active site. The mechanism of "O(2) tolerance" involves a critical role for the Fe-S clusters of the electron relay, which is to ensure the availability-for immediate transfer back to the active site-of all of the electrons required to reduce an attacking O(2) molecule completely to harmless H(2)O. The unique [4Fe-3S] cluster proximal to the active site is crucial because it can rapidly transfer two of the electrons needed. Here we investigate and establish the equally crucial role of the high potential medial [3Fe-4S] cluster, located >20 Å from the active site. A variant, P242C, in which the medial [3Fe-4S] cluster is replaced by a [4Fe-4S] cluster, is unable to sustain steady-state H(2) oxidation activity in 1% O(2). The [3Fe-4S] cluster is essential only for the first stage of complete O(2) reduction, ensuring the supply of all three electrons needed to form the oxidized inactive state "Ni-B" or "Ready" (Ni(III)-OH). Potentiometric titrations show that Ni-B is easily reduced (E(m) ≈ +0.1 V at pH 6.0); this final stage of the O(2)-tolerance mechanism regenerates active enzyme, effectively completing a competitive four-electron oxidase cycle and is fast regardless of alterations at the proximal or medial clusters. As a consequence of all these factors, the enzyme's response to O(2), viewed by its electrocatalytic activity in protein film electrochemistry (PFE) experiments, is merely to exhibit attenuated steady-state H(2) oxidation activity; thus, O(2) behaves like a reversible inhibitor rather than an agent that effectively causes irreversible inactivation. The data consolidate a rich picture of the versatile role of Fe-S clusters in electron relays and suggest that Hyd-1 can function as a proficient hydrogen oxidase. PMID

  7. Genetic analysis of strawberry fruit aroma and identification of O-methyltransferase FaOMT as the locus controlling natural variation in mesifurane content.

    PubMed

    Zorrilla-Fontanesi, Yasmín; Rambla, José-Luis; Cabeza, Amalia; Medina, Juan J; Sánchez-Sevilla, José F; Valpuesta, Victoriano; Botella, Miguel A; Granell, Antonio; Amaya, Iraida

    2012-06-01

    Improvement of strawberry (Fragaria × ananassa) fruit flavor is an important goal in breeding programs. To investigate genetic factors controlling this complex trait, a strawberry mapping population derived from genotype '1392', selected for its superior flavor, and '232' was profiled for volatile compounds over 4 years by headspace solid phase microextraction coupled to gas chromatography and mass spectrometry. More than 300 volatile compounds were detected, of which 87 were identified by comparison of mass spectrum and retention time to those of pure standards. Parental line '1392' displayed higher volatile levels than '232', and these and many other compounds with similar levels in both parents segregated in the progeny. Cluster analysis grouped the volatiles into distinct chemically related families and revealed a complex metabolic network underlying volatile production in strawberry fruit. Quantitative trait loci (QTL) detection was carried out over 3 years based on a double pseudo-testcross strategy. Seventy QTLs covering 48 different volatiles were detected, with several of them being stable over time and mapped as major QTLs. Loci controlling γ-decalactone and mesifurane content were mapped as qualitative traits. Using a candidate gene approach we have assigned genes that are likely responsible for several of the QTLs. As a proof of concept we show that one homoeolog of the O-methyltransferase gene (FaOMT) is the locus responsible for the natural variation of mesifurane content. Sequence analysis identified 30 bp in the promoter of this FaOMT homoeolog containing putative binding sites for basic/helix-loop-helix, MYB, and BZIP transcription factors. This polymorphism fully cosegregates with both the presence of mesifurane and the high expression of FaOMT during ripening. PMID:22474217

  8. NEW CHEMICAL ALTERNATIVE FOR OZONE-DEPLETING SUBSTANCES: HFC-236FA

    EPA Science Inventory

    The report gives results of a preliminary evaluation of a new hydrofluorocarbon (HFC)--HFC-236fa or 1,1,1,3,3,3-hexafluoropropane--as a possible alternative for chlorofluorocarbon (CFC)-114 (1,2-dichloro-1,1,2,2-tetrafluoroethane) refrigerant for chillers and as a possible fire s...

  9. HEAT TRANSFER EVALUATION OF HFC-236FA IN CONDENSATION AND EVAPORATION

    EPA Science Inventory

    The report gives results of an evaluation of the shell-side heat transfer performance of hydrofluorocarbon (HFC)-236fa, which is considered to be a potential substitute for chlorofluorocarbon (CFC)-114 in Navy shipboard chillers, for both conventional finned [1024- and 1575-fpm (...

  10. Exterior building details of Building C, east façade: historic fouroverfour ...

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

    Exterior building details of Building C, east façade: historic four-over-four window, brick lintel, brick quoins, corbelled brick cornice, spiral metal staircase to inclined stairs rising to second floor cantilever wooden walkway; westerly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  11. Exterior building details of Building B, west façade: road level ...

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

    Exterior building details of Building B, west façade: road level four-over-four double-hung painted-wood windows with brick sill and arch brick lintels; easterly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  12. Exterior building details of Building C, east façade: inscribed date ...

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

    Exterior building details of Building C, east façade: inscribed date panel "hospital 1885", corbelled brick belt course, parapet, second floor historic four-over-four window with brick lintels, quoins and decorative metal grilled, cantilever wooden walkway; westerly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  13. Exterior building details of Building A; west façade: exposed common ...

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

    Exterior building details of Building A; west façade: exposed common bond brick wall, arched brick lintels over a two single-light casement window with brick sills, arched brick lintel over door cornice; easterly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  14. Exterior building details of Building C, south façade: second floor" ...

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

    Exterior building details of Building C, south façade: second floor" four-over-four windows, arch brick lintels, brick sills, decorative metal grilles and tiebacks; northwesterly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  15. Exterior building details of Building C, west façade: second floor: ...

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

    Exterior building details of Building C, west façade: second floor: four-over-four windows, arch brick lintels, brick sills, decorative metal grilles; easterly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  16. Exterior building details of Building B, west façade: two paintedwood ...

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

    Exterior building details of Building B, west façade: two painted-wood single-light casements over two-light casements with concrete sill and arch brick lintel, over infilled brick patch with arch brick lintel, brick lintel above windows and brick infilled oval; easterly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  17. DAO spectroscopic classification of ASASSN-16fa = AT 2016cct in UGC 6434

    NASA Astrophysics Data System (ADS)

    Balam, D. D.; Graham, M. L.

    2016-05-01

    A spectrum of ASASSN-16fa = 2016cct (ATEL #9045) was obtained using the 1.82-m Plaskett telescope (National Research Council of Canada) covering the range 375-710 nm (resolution 0.35 nm) on UT May 13.26 that reveals the object to be a type-Ia (normal) supernova near maximum light.

  18. Exterior building details of Building A; east façade: recessed panel ...

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

    Exterior building details of Building A; east façade: recessed panel inscribed "1859", historic window opening with concrete sill above door, cement plaster dentil course and cornice, truncated wood beam ends, plaster finished brick wall, granite base; westerly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  19. Exterior building details of Building C, east façade: brick quoins, ...

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

    Exterior building details of Building C, east façade: brick quoins, brick lintels, brick window sills, decorative metal grilles, scored cement finished brick wall; westerly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  20. Exterior building details of Building A; east façade: profiled cement ...

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

    Exterior building details of Building A; east façade: profiled cement plaster door surround, black mesh gate protects a two-light transom atop non-original metal door; westerly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  1. Exterior building details of Building A; east façade: concrete staircase, ...

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

    Exterior building details of Building A; east façade: concrete staircase, profiled cement, plaster door surround, recessed panel inscribed "1859", historic window opening with concrete sill above door, cement plaster dentil course and cornice truncated wood beam ends, plaster finished brick wall, granite base; westerly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  2. Exterior building details of Building C, east façade: historic six ...

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

    Exterior building details of Building C, east façade: historic six light entry double door with three light transom, historic six light door with a one light transom, arch brick lintels and quoins, scored cement plaster finished brick walls; westerly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  3. MISCIBILITY, SOLUBILITY, VISCOSITY, AND DENSITY MEASUREMENTS FOR R-236FA WITH POTENTIAL LUBRICANTS

    EPA Science Inventory

    The report gives results of miscibility, solubility, viscosity, and density measurements for refrigerant R-236fa and two potential lubricants . (The data are needed to determine the suitability of refrigerant/lubricant combinations for use in refrigeration systems.) The tested oi...

  4. Exterior building details of Building A; north façade: two threelight ...

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

    Exterior building details of Building A; north façade: two three-light wood casement windows flank a three-light fixed wood window with concrete sill; southerly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  5. Exterior building details of Building A; east façade: fixed fiveoverfive ...

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

    Exterior building details of Building A; east façade: fixed five-over-five wood windows with five-light hoppers with concrete sills; westerly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  6. Automatic modelling of building façade objects via primitive shapes

    NASA Astrophysics Data System (ADS)

    Hetti Arachchige, N.; Perera, S.

    2014-08-01

    This paper presents a new approach to recognize individual façade objects and to reconstruct such objects in 3D using MLS point clouds. Core of the approach is a primitive shape based algorithm, which introduces building primitives, to identify the façade objects separately from other irrelevant objects and then to model the correct topology. The primitive shape is identified against defined different primitive shapes by using the Douglas-Peucker algorithm. The advantage of this process is that it offers an ability not only to model correct geometric shapes but also to remove occlusion effects from the final model. To evaluate the validity of the proposed approach, experiments have been conducted using two types of street scene point clouds captured by Optech Lynx Mobile Mapper System and Z+F laser scanner. Results of the experiments show that the completeness, correctness, and quality of the reconstructed building façade objects are well over 90 %, proving the proposed method is a promising solution for modelling 3D façade objects with different geometric shapes.

  7. Detail, typical window fenestrations, east façade, view to westnorthwest (135mm ...

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

    Detail, typical window fenestrations, east façade, view to west-northwest (135mm lens). Note "timber" header, adobe bench in front of window. - Burton Park, Club House & Amphitheater, Adjacent ot south end of Chestnut Avenue, San Carlos, San Mateo County, CA

  8. Genome Sequence of the Alkaline-Tolerant Cellulomonas sp. Strain FA1

    DOE PAGESBeta

    Cohen, Michael F.; Hu, Ping; Nguyen, My Vu; Kamennaya, Nina; Brown, Natasha; Woyke, Tanja; Kyrpides, Nikos; Holman, Hoi-Ying; Torok, Tamas

    2015-06-18

    We present the genome of the cellulose-degrading Cellulomonas sp. strain FA1 isolated from an actively serpentinizing highly alkaline spring. Knowledge of this genome will enable studies into the molecular basis of plant material degradation in alkaline environments and inform the development of lignocellulose bioprocessing procedures for biofuel production.

  9. Expression of crylFa in bahiagrass enhances resistance to fall armyworm

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bahiagrass (Paspalum notatum var. Flugge) is the predominant forage grass in Florida and in subtropical regions worldwide. To improve pest resistance against fall armyworm (Spodoptera frugiperda J. E. Smith), an optimized cry1Fa gene encoding a '-endotoxin from Bacillus thuringiensis was synthesized...

  10. Genome Sequence of the Alkaline-Tolerant Cellulomonas sp. Strain FA1

    SciTech Connect

    Cohen, Michael F.; Hu, Ping; Nguyen, My Vu; Kamennaya, Nina; Brown, Natasha; Woyke, Tanja; Kyrpides, Nikos; Holman, Hoi-Ying; Torok, Tamas

    2015-06-18

    We present the genome of the cellulose-degrading Cellulomonas sp. strain FA1 isolated from an actively serpentinizing highly alkaline spring. Knowledge of this genome will enable studies into the molecular basis of plant material degradation in alkaline environments and inform the development of lignocellulose bioprocessing procedures for biofuel production.

  11. HEAT TRANSFER EVALUATION OF HFC-236FA IN CONDENSATION AND EVAPORATION

    EPA Science Inventory

    The report gives results of an evaluation of the shell-side heat transfer performance of hydrofluorocarbon (HFC)-236fa, which is considered to be a potential substitute for chlorofluorocarbon (CFC)-114 in Navy shipboard chillers, for both conventional finned (1024- and 1575-fpm (...

  12. View of rear façade of office building; note projecting bay, ...

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

    View of rear façade of office building; note projecting bay, above the basement level, which commanded a view of the iron works - Everett Iron Company, Office Building, 0.25 mile Southwest of Everett, Earlston, Bedford County, PA

  13. Leaching of biocides from façades under natural weather conditions.

    PubMed

    Burkhardt, M; Zuleeg, S; Vonbank, R; Bester, K; Carmeliet, J; Boller, M; Wangler, T

    2012-05-15

    Biocides are included in organic building façade coatings as protection against biological attack by algae and fungi but have the potential to enter the environment via leaching into runoff from wind driven rain. The following field study correlates wind driven rain to runoff and measured the release of several commonly used organic biocides (terbutryn, Irgarol 1051, diuron, isoproturon, OIT, DCOIT) in organic façade coatings from four coating systems. During one year of exposure of a west oriented model house façade in the Zurich, Switzerland area, an average of 62.7 L/m(2), or 6.3% of annual precipitation came off the four façade panels installed as runoff. The ISO method for calculating wind driven rain loads is adapted to predict runoff and can be used in the calculation of emissions in the field. Biocide concentrations tend to be higher in the early lifetime of the coatings and then reach fairly consistent levels later, generally ranging on the order of mg/L or hundreds of μg/L. On the basis of the amount remaining in the film after exposure, the occurrence of transformation products, and the calculated amounts in the leachate, degradation plays a significant role in the overall mass balance. PMID:22524149

  14. Missionaries and Tonic Sol-fa Music Pedagogy in 19th-Century China

    ERIC Educational Resources Information Center

    Southcott, Jane E.; Lee, Angela Hao-Chun

    2008-01-01

    In the 19th century, Christian missionaries in China, as elsewhere, used the Tonic Sol-fa method of music instruction to aid their evangelizing. This system was designed to improve congregational singing in churches, Sunday schools and missions. The London Missionary Society and other evangelical groups employed the method. These missionaries took…

  15. An adaptive cognitive dissociation between willingness to help kin and nonkin in Samoan Fa'afafine.

    PubMed

    Vasey, Paul L; VanderLaan, Doug P

    2010-02-01

    Androphilia refers to sexual attraction and arousal to adult males, whereas gynephilia refers to sexual attraction and arousal to adult females. Previous research has demonstrated that Samoan male androphiles (known locally as fa'afafine) exhibit significantly higher altruistic tendencies toward nieces and nephews than do Samoan women and gynephilic men. The present study examined whether adaptive design features characterize the psychological mechanisms underlying fa'afafine's elevated avuncular tendencies. The association between altruistic tendencies toward nieces and nephews and altruistic tendencies toward nonkin children was significantly weaker among fa'afafine than among Samoan women and gynephilic men. We argue that this cognitive dissociation would allow fa'afafine to allocate resources to nieces and nephews in a more economical, efficient, reliable, and precise manner. These findings are consistent with the kin selection hypothesis, which suggests that androphilic males have been selected over evolutionary time to act as "helpers-in-the-nest," caring for nieces and nephews and thereby increasing their own indirect fitness. PMID:20424059

  16. Doppler global velocimetry measurements of the vortical flow above an F/A-18

    NASA Astrophysics Data System (ADS)

    Lee, Joseph W.; Meyers, James F.; Cavone, Angelo A.; Suzuki, Karen E.

    1993-01-01

    A Doppler global velocimeter is employed to study the vortical flow above an F/A-18 at 25 degree angle of attack. The measurements indicate that the flow possessed the same characteristics as the vortical flow above a standard delta wing. Image to image comparisons clearly indicate that the flow striking the vertical stabilizers is not chaotic.

  17. Slicing Method for curved façade and window extraction from point clouds

    NASA Astrophysics Data System (ADS)

    Iman Zolanvari, S. M.; Laefer, Debra F.

    2016-09-01

    Laser scanning technology is a fast and reliable method to survey structures. However, the automatic conversion of such data into solid models for computation remains a major challenge, especially where non-rectilinear features are present. Since, openings and the overall dimensions of the buildings are the most critical elements in computational models for structural analysis, this article introduces the Slicing Method as a new, computationally-efficient method for extracting overall façade and window boundary points for reconstructing a façade into a geometry compatible for computational modelling. After finding a principal plane, the technique slices a façade into limited portions, with each slice representing a unique, imaginary section passing through a building. This is done along a façade's principal axes to segregate window and door openings from structural portions of the load-bearing masonry walls. The method detects each opening area's boundaries, as well as the overall boundary of the façade, in part, by using a one-dimensional projection to accelerate processing. Slices were optimised as 14.3 slices per vertical metre of building and 25 slices per horizontal metre of building, irrespective of building configuration or complexity. The proposed procedure was validated by its application to three highly decorative, historic brick buildings. Accuracy in excess of 93% was achieved with no manual intervention on highly complex buildings and nearly 100% on simple ones. Furthermore, computational times were less than 3 sec for data sets up to 2.6 million points, while similar existing approaches required more than 16 hr for such datasets.

  18. Antifungal efficiency assessment of the TiO2 coating on façade paints.

    PubMed

    Vučetić, Snežana B; Rudić, Ognjen Lj; Markov, Siniša L; Bera, Oskar J; Vidaković, Ana M; Skapin, Andrijana S Sever; Ranogajec, Jonjaua G

    2014-10-01

    The work studies the photocatalytic activity and the antifungal efficiency of the TiO2/Zn-Al coatings placed on the target commercial façade paints. The photocatalytic active nanocomposite based on TiO2 and Zn-Al-layered double hydroxides (ZnAl-LDHs) was synthesized by a wet impregnation technique with 3 % w/w TiO2. The freshly prepared suspension was applied by spray technique on the surfaces of the white façade paints. The goal of the work was to develop a method that quickly quantifies the antifungal activity of the commercial façade paints with and without biocidal components covered with a photocatalytic coating. The essence of the proposed method is the monitoring of the fungal growth (artificial ageing conditions) and the quantification of its development (UV-A 0.13 mWcm(-2)) on the façade paint surfaces. A special fungus nutrient (potato dextrose agar (PDA)) was inoculated with the spores of the Aspergillus niger ATCC 6275, and the test samples (façade paints with and without photocatalytic coating) were placed on the inoculated nutrient in the petri dishes. The images of the fungal growth on the samples of the facade paints, during a period of 5 days, were imported into Matlab R2012a where they were converted to binary images (BW), based on the adequate threshold. The percentage of the surface coverage was calculated by applying the specifically written program code which determines the ratio of the black and white pixels. The black pixels correspond to the surface covered with hyphae and mycelia of the fungus. PMID:24875311

  19. Flow boiling heat transfer of R134a, R236fa and R245fa in a horizontal 1.030 mm circular channel

    SciTech Connect

    Ong, Chin L.; Thome, John R.

    2009-04-15

    This research focuses on acquiring accurate flow boiling heat transfer data and flow pattern visualization for three refrigerants, R134a, R236fa and R245fa in a 1.030 mm channel. We investigate trends in the data, and their possible mechanisms, for mass fluxes from 200 to 1600 kg/m{sup 2}s, heat fluxes from 2.3 kW/m{sup 2} to 250 kW/m{sup 2} at T{sub sat} = 31 C and {delta}T{sub sub} from 2 to 9 K. The local saturated flow boiling heat transfer coefficients display a heat flux and a mass flux dependency but no residual subcooling influence. The changes in heat transfer trends correspond well with flow regime transitions. These were segregated into the isolated bubble (IB) regime, the coalescing bubble (CB) regime, and the annular (A) regime for the three fluids. The importance of nucleate boiling and forced convection in these small channels is still relatively unclear and requires further research. (author)

  20. An Evaluation Technique for an F/A-18 Aircraft Loads Model Using F/A-18 Systems Research Aircraft Flight Data

    NASA Technical Reports Server (NTRS)

    Olney, Candida D.; Hillebrandt, Heather; Reichenbach, Eric Y.

    2000-01-01

    A limited evaluation of the F/A-18 baseline loads model was performed on the Systems Research Aircraft at NASA Dryden Flight Research Center (Edwards, California). Boeing developed the F/A-18 loads model using a linear aeroelastic analysis in conjunction with a flight simulator to determine loads at discrete locations on the aircraft. This experiment was designed so that analysis of doublets could be used to establish aircraft aerodynamic and loads response at 20 flight conditions. Instrumentation on the right outboard leading edge flap, left aileron, and left stabilator measured the hinge moment so that comparisons could be made between in-flight-measured hinge moments and loads model-predicted values at these locations. Comparisons showed that the difference between the loads model-predicted and in-flight-measured hinge moments was up to 130 percent of the flight limit load. A stepwise regression technique was used to determine new loads derivatives. These derivatives were placed in the loads model, which reduced the error to within 10 percent of the flight limit load. This paper discusses the flight test methodology, a process for determining loads coefficients, and the direct comparisons of predicted and measured hinge moments and loads coefficients.

  1. Analysis of Tangential Slot Blowing on F/A-18 Isolated Forebody

    NASA Technical Reports Server (NTRS)

    Gee, Ken; Rizk, Yehia M.; Schiff, Lewis B.

    1995-01-01

    The generation of significant side forces and yawing moments on an F/A-18 fuselage through tangential slot blowing is analyzed using computational fluid dynamics. The effects of freestream Mach number, jet exit conditions, jet length, and jet location are studied. The effects of over- and underblowing on force and moment production are analyzed. Non-time-accurate solutions are obtained to determine the steady-state side forces, yawing moments, and surface pressure distributions generated by tangential slot blowing. Time-accurate solutions are obtained to study the force onset time lag of tangential slot blowing. Comparison with available experimental data from full-scale wind-tunnel and subscale wind-tunnel tests are made. This computational analysis complements the experimental results and provides a detailed understanding of the effects of tangential slot blowing on the flowfield about the isolated F/A-18 forebody. Additionally, it extends the slot-blowing database to transonic maneuvering Mach numbers.

  2. Operation record of the large 9FA based CCGTs built in Europe

    SciTech Connect

    Pfleger, D.

    1998-07-01

    GEC ALSTHOM references for 9F/FA based Combined Cycles in operation in Europe have accumulated more than 170,000 hours (mid-April 98), a record for plants of their size with advanced gas turbines. Based on a standardized and optimized design, the single shaft concept has demonstrated its high availability, its operating capabilities with short start-up times and adaptation to various site-related cooling systems without any impact on the overall operational capabilities. These experiences have confirmed the suitability of the VEGA 109FA SS 3PRH for today's customer requirements in terms of flexible operation and ease of maintenance, high cycle efficiency, high reliability and short lead times for a minimum life cycle cost.

  3. Navier-Stokes solutions about the F/A-18 forebody-LEX configuration

    NASA Technical Reports Server (NTRS)

    Ghaffari, Farhad; Luckring, James M.; Thomas, James L.; Bates, Brent L.

    1989-01-01

    Three-dimensional viscous flow computations are presented for the F/A-18 forebody-LEX (Leading Edge EXtensions) geometry. Solutions are obtained from an algorithm for the compressible Navier-Stokes equations which incorporates an upwind-biased, flux-difference-splitting approach along with longitudinally-patched grids. Results are presented for both laminar and fully turbulent flow assumptions and include correlations with wind tunnel as well as flight-test results. A good quantitative agreement for the forebody surface pressure distribution is achieved between the turbulent computations and wind tunnel measurements at Mach number 0.6. The computed turbulent surface flow patterns on the forebody qualitatively agree well with in-flight surface flow patterns obtained on an F/A-18 aircraft at Mach number 0.34.

  4. Life extension of F/A 18 intake nacelle using add-on dampers

    NASA Astrophysics Data System (ADS)

    Chiu, W. K.; Siovitz, U.; Galea, Stephen C.; Koss, Leonard L.

    1998-06-01

    Acoustic fatigue is due to very high intensity excitation as a result of pressure waves caused by either engine/or aerodynamic effects. Currently a large portion of the F/A-18 fleet has suffered from acoustic fatigue cracking in skin panel on the lower external surface of the inlet nacelle. There has been a long history of cracking in this region, which is attributed to fatigue caused by acoustic excitation of the panel during the operation of the aircraft. Efforts to alleviate these fatigue crack growth with traditional boron/epoxy patches was not successful. This paper seek to select a set of damping material suitable for the reinforcement/damping treatment to alleviate the acoustic fatigue damage of the nacelle panel of the F/A-18 aircraft. This experimental study will address the relevance of constrained layer damping reducing the incidence of acoustic fatigue problems in the nacelle region.

  5. Exterior building details of Building A; west façade: white painted ...

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

    Exterior building details of Building A; west façade: white painted brick wall of road and second level, road level: paired four-light casement window and a small single-light wood casement window; second level: four-over-four wood double-hung window and a six-light horizontal pivot over a three-light fixed window; easterly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  6. Exterior building details of Building B, east façade: embedded wood ...

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

    Exterior building details of Building B, east façade: embedded wood beams and interrupted dentil course cornice resulting from the removal of the third floor tuberculosis ward, yard level paneled Dutch door, second level two a typical six-light wood casement windows over a single-panel wood door with four light exits to fire escape; westerly view - San Quentin State Prison, Building 22, Point San Quentin, San Quentin, Marin County, CA

  7. Full-Scale Wind Tunnel Studies of F/A-18 Tail Buffet

    NASA Technical Reports Server (NTRS)

    Meyn, Larry A.; James, Kevin D.

    1993-01-01

    Tail buffet studies were conducted on a full-scale, production, F/A-18, fighter aircraft in the 80- by 120-Foot Wind Tunnel of the National Full-Scale Aerodynamic Complex at NASA Ames Research Center in Moffett Field, California. The F/A-18 was tested over an angle-of-attack range of 18deg to 50deg, a side-slip range of -15deg to 15deg, and at wind speeds of up to 100 knots. The maximum speed corresponds to a Reynolds number of 12.3 x 10(exp 6) based on mean aerodynamic chord and a Mach number of 0.15. The port, vertical tail fin was instrumented with thirty-two surface pressure transducers, arranged in four by four arrays on both sides on the fin. The aircraft was also equipped with a removable Leading Edge eXtension (LEX) fence that is used on F/A-18 aircraft to reduce tail buffet loads. Time-averaged, power-spectral analysis results are presented for the tail fin bending moment derived from the integrated pressure field. The results are only for the zero side-slip condition, both with and without the LEX fence. The LEX fence significantly reduces the magnitude of the root-mean-square pressures and bending moments. Scaling issues are addressed by comparing full-scale results for pressures at the 60%-span and 45%-chord location with published results of small-scale, F/A-18 tail-buffet tests. The comparison shows that the tail buffet frequency scales very well with length and velocity. Root-mean-square pressures and power spectra do not scale as well. The LEX fence is shown to reduce tail buffet loads at all model scales.

  8. Sharing Facilities and Administrative (F&A) Cost Recovery to Facilitate Interdisciplinary Research

    PubMed Central

    Kulage, Kristine M.; Larson, Elaine L.; Begg, Melissa D.

    2011-01-01

    Purpose Despite increasing interest in interdisciplinary research, researchers consistently cite institutional barriers as deterrents. Researchers, administrators, and others have suggested developing processes for sharing facilities and administrative (F&A) cost recovery as one way to support collaborative research. Therefore, the authors reviewed current policies for sharing F&A cost recovery and user satisfaction with them. Method In 2010, through reviewing institutional Websites and surveying researchers and grants administrators from a range of institutions, the authors identified a number of different policies currently employed and assessed user satisfaction with them. Results While most respondents (80.7%, 205/254) agreed that a standard policy for sharing F&A cost recovery would facilitate interdisciplinary collaboration, only 35.4% (90/254) reported that their institutions had such a policy. Among the 85 respondents whose institutions had a policy, most (66 [77.6%]) reported that the policy applied to grants with multiple principal investigators or co-investigators across departments or schools, and 68 (80.0%) reported satisfaction with the policy. Respondents from institutions with policies were significantly more likely to endorse the notion that policies are helpful compared to those who reported that their institutions did not have such policies or were unsure of their existence (89% versus 76%, P = 0.014). The authors detected no significant differences in satisfaction scores based on type of policy, whether determined by investigator effort, space allocation, or other considerations (P = 0.29). Conclusions These data support the need for institutions to establish formal policies for sharing F&A cost recovery as a way to promote interdisciplinary research collaboration. PMID:21248610

  9. Wavelet Analyses of F/A-18 Aeroelastic and Aeroservoelastic Flight Test Data

    NASA Technical Reports Server (NTRS)

    Brenner, Martin J.

    1997-01-01

    Time-frequency signal representations combined with subspace identification methods were used to analyze aeroelastic flight data from the F/A-18 Systems Research Aircraft (SRA) and aeroservoelastic data from the F/A-18 High Alpha Research Vehicle (HARV). The F/A-18 SRA data were produced from a wingtip excitation system that generated linear frequency chirps and logarithmic sweeps. HARV data were acquired from digital Schroeder-phased and sinc pulse excitation signals to actuator commands. Nondilated continuous Morlet wavelets implemented as a filter bank were chosen for the time-frequency analysis to eliminate phase distortion as it occurs with sliding window discrete Fourier transform techniques. Wavelet coefficients were filtered to reduce effects of noise and nonlinear distortions identically in all inputs and outputs. Cleaned reconstructed time domain signals were used to compute improved transfer functions. Time and frequency domain subspace identification methods were applied to enhanced reconstructed time domain data and improved transfer functions, respectively. Time domain subspace performed poorly, even with the enhanced data, compared with frequency domain techniques. A frequency domain subspace method is shown to produce better results with the data processed using the Morlet time-frequency technique.

  10. Development and characterization of a high sensitivity segmented Fast Neutron Spectrometer (FaNS-2)

    NASA Astrophysics Data System (ADS)

    Langford, T. J.; Beise, E. J.; Breuer, H.; Heimbach, C. R.; Ji, G.; Nico, J. S.

    2016-01-01

    We present the development of a segmented fast neutron spectrometer (FaNS-2) based upon plastic scintillator and 3He proportional counters. It was designed to measure both the flux and spectrum of fast neutrons in the energy range of few MeV to 1 GeV. FaNS-2 utilizes capture-gated spectroscopy to identify neutron events and reject backgrounds. Neutrons deposit energy in the plastic scintillator before capturing on a 3He nucleus in the proportional counters. Segmentation improves neutron energy reconstruction while the large volume of scintillator increases sensitivity to low neutron fluxes. A main goal of its design is to study comparatively low neutron fluxes, such as cosmogenic neutrons at the Earth's surface, in an underground environment, or from low-activity neutron sources. In this paper, we present details of its design and construction as well as its characterization with a calibrated 252Cf source and monoenergetic neutron fields of 2.5 MeV and 14 MeV. Detected monoenergetic neutron spectra are unfolded using a Singular Value Decomposition method, demonstrating a 5% energy resolution at 14 MeV. Finally, we discuss plans for measuring the surface and underground cosmogenic neutron spectra with FaNS-2.

  11. Full-Scale Wind-Tunnel Studies of F/A-18 Tail Buffet

    NASA Technical Reports Server (NTRS)

    Meyn, Larry A.; James, Kevin D.

    1996-01-01

    Tail buffet studies were conducted on a full-scale, production F/A-18 fighter aircraft in the 80 by 120 ft Wind Tunnel at NASA Ames Research Center. The F/A-18 was tested over an angle-of-attack range of 18-50 deg, and at wind speeds of up to 168 ft/s, corresponding to a Reynolds number of 12.3x10(exp 6) based on mean aerodynamic chord and a Mach number of 0.15. The port, vertical tail fin was instrumented and the aircraft was equipped with a removable leading-edge extension (LEX) fence. Time-averaged, power-spectral analysis results are presented for the tail fin bending moment derived from the integrated pressure field, for the zero side-slip condition, both with and without the LEX fence. The LEX fence significantly reduces the magnitude of the rms pressures and bending moments. Scaling issues are addressed by comparing full-scale results for pressures at the 60%-span and 45%-chord location with small-scale, F/A-18 tail-buffet data. The comparison shows that the tail buffet frequency scales very well with length and velocity. Root-mean-square pressures and power spectra do not scale as well. The LEX fence is shown to reduce tail buffet loads at all model scales.

  12. Aeroservoelastic Modeling and Validation of a Thrust-Vectoring F/A-18 Aircraft

    NASA Technical Reports Server (NTRS)

    Brenner, Martin J.

    1996-01-01

    An F/A-18 aircraft was modified to perform flight research at high angles of attack (AOA) using thrust vectoring and advanced control law concepts for agility and performance enhancement and to provide a testbed for the computational fluid dynamics community. Aeroservoelastic (ASE) characteristics had changed considerably from the baseline F/A-18 aircraft because of structural and flight control system amendments, so analyses and flight tests were performed to verify structural stability at high AOA. Detailed actuator models that consider the physical, electrical, and mechanical elements of actuation and its installation on the airframe were employed in the analysis to accurately model the coupled dynamics of the airframe, actuators, and control surfaces. This report describes the ASE modeling procedure, ground test validation, flight test clearance, and test data analysis for the reconfigured F/A-18 aircraft. Multivariable ASE stability margins are calculated from flight data and compared to analytical margins. Because this thrust-vectoring configuration uses exhaust vanes to vector the thrust, the modeling issues are nearly identical for modem multi-axis nozzle configurations. This report correlates analysis results with flight test data and makes observations concerning the application of the linear predictions to thrust-vectoring and high-AOA flight.

  13. Flight Test of the F/A-18 Active Aeroelastic Wing Airplane

    NASA Technical Reports Server (NTRS)

    Voracek, David

    2007-01-01

    A viewgraph presentation of flight tests performed on the F/A active aeroelastic wing airplane is shown. The topics include: 1) F/A-18 AAW Airplane; 2) F/A-18 AAW Control Surfaces; 3) Flight Test Background; 4) Roll Control Effectiveness Regions; 5) AAW Design Test Points; 6) AAW Phase I Test Maneuvers; 7) OBES Pitch Doublets; 8) OBES Roll Doublets; 9) AAW Aileron Flexibility; 10) Phase I - Lessons Learned; 11) Control Law Development and Verification & Validation Testing; 12) AAW Phase II RFCS Envelopes; 13) AAW 1-g Phase II Flight Test; 14) Region I - Subsonic 1-g Rolls; 15) Region I - Subsonic 1-g 360 Roll; 16) Region II - Supersonic 1-g Rolls; 17) Region II - Supersonic 1-g 360 Roll; 18) Region III - Subsonic 1-g Rolls; 19) Roll Axis HOS/LOS Comparison Region II - Supersonic (open-loop); 20) Roll Axis HOS/LOS Comparison Region II - Supersonic (closed-loop); 21) AAW Phase II Elevated-g Flight Test; 22) Region I - Subsonic 4-g RPO; and 23) Phase II - Lessons Learned

  14. Active Aeroelastic Wing Aerodynamic Model Development and Validation for a Modified F/A-18A

    NASA Technical Reports Server (NTRS)

    Cumming, Stephen B.; Diebler, Corey G.

    2005-01-01

    A new aerodynamic model has been developed and validated for a modified F/A-18A used for the Active Aeroelastic Wing (AAW) research program. The goal of the program was to demonstrate the advantages of using the inherent flexibility of an aircraft to enhance its performance. The research aircraft was an F/A-18A with wings modified to reduce stiffness and a new control system to increase control authority. There have been two flight phases. Data gathered from the first flight phase were used to create the new aerodynamic model. A maximum-likelihood output-error parameter estimation technique was used to obtain stability and control derivatives. The derivatives were incorporated into the National Aeronautics and Space Administration F-18 simulation, validated, and used to develop new AAW control laws. The second phase of flights was used to evaluate the handling qualities of the AAW aircraft and the control law design process, and to further test the accuracy of the new model. The flight test envelope covered Mach numbers between 0.85 and 1.30 and dynamic pressures from 600 to 1250 pound-force per square foot. The results presented in this report demonstrate that a thorough parameter identification analysis can be used to improve upon models that were developed using other means. This report describes the parameter estimation technique used, details the validation techniques, discusses differences between previously existing F/A-18 models, and presents results from the second phase of research flights.

  15. Beyond third generation: a sensor-fusion targeting FLIR pod for the F/A-18

    NASA Astrophysics Data System (ADS)

    Krebs, William K.; Scribner, Dean A.; Miller, Geoffrey M.; Ogawa, James S.; Schuler, Jonathon M.

    1998-03-01

    The Navy and Marine Corps F/A-18 pilots state that the targeting FLIR system does not provide enough target definition and clarity. As a result, high altitude tactics missions are the most difficult due to the limited amount of time available to identify the target. If the targeting FLIR system had a better stand-off range and an improved target contrast then the pilots' task would be easier. Unfortunately, the replacement cost of the existing FLIR equipment is prohibitive. The purpose of this study is to modify the existing F/A-18 targeting FLIR system with a dual-band color sensor to improve target contrast and stand- off ranges. Methods: A non-real-time color sensor fusion system was flown on a NASA F/A-18 in a NITE Hawk targeting FLIR pod. Flight videotape was recorded from a third generation image intensified CCD and a first generation long-wave infrared sensor. A standard visual search task was used to assess whether pilots' situational awareness was improved by combining the two sensor videotape sequences into a single fused color or grayscale representation. Results: Fleet aviators showed that color fusion improved target detection, but hindered situational awareness. Aviators reported the lack of color constancy caused the scene to be unaesthetically pleasing; however, target detection was enhanced. Conclusion: A color fusion scene may benefit targeting applications but hinder situational awareness.

  16. Effects of carbon dioxide level (PCO sub 2 ) on the fibrinolytic activity (FA) of pulmonary artery endothelial cells (PAEC)

    SciTech Connect

    Langleben, D.; Moroz, L.A.; Danes, D. )

    1990-02-26

    Recovery from pulmonary thromboembolism depends on the rapidity and completeness of clot lysis. This involves endogenous fibrinolytic mechanisms, particularly the balance between plasminogen activators and inhibitors produced by endothelial cells. Hypocapnia is common in pulmonary embolism, however it is not known if endothelial fibrinolytic function is affected by PCO{sub 2}. The authors therefore measured the FA in medium (MCDB-131, 0.5% albumin) conditioned for 20 hours in-vitro by exposure to confluent cultures of bovine proximal PAEC. During conditioning, cells were exposed to 5% CO{sub 2} in air (PCO{sub 2} - 36-40mm Hg, CONTROL), or various PCO{sub 2} levels (30-55 mmHg, in air). FA of conditioned medium was determined by {sup 125}I-fibrin solid phase assay, with addition of plasminogen (10 ug/ml). With PCO{sub 2} levels {le} 35 mmHg, FA in the conditioned medium was 5 to 18% higher than CONTROL FA. When PCO{sub 2} was {ge} 45 mmHg, FA decreased 5 to 60% as compared to CONTROL FA. There was a significant negative linear relationship between PCO{sub 2} and FA. Thus, PCO{sub 2} level can affect PAEC mediated plasminogen activation. This finding may be relevant to in-vivo clearance of clots from pulmonary arteries.

  17. Differential Protection of Cry1Fa Toxin against Spodoptera frugiperda Larval Gut Proteases by Cadherin Orthologs Correlates with Increased Synergism

    PubMed Central

    Rahman, Khalidur; Abdullah, Mohd Amir F.; Ambati, Suresh; Taylor, Milton D.

    2012-01-01

    The Cry proteins produced by Bacillus thuringiensis (Bt) are the most widely used biopesticides effective against a range of crop pests and disease vectors. Like chemical pesticides, development of resistance is the primary threat to the long-term efficacy of Bt toxins. Recently discovered cadherin-based Bt Cry synergists showed the potential to augment resistance management by improving efficacy of Cry toxins. However, the mode of action of Bt Cry synergists is thus far unclear. Here we elucidate the mechanism of cadherin-based Cry toxin synergism utilizing two cadherin peptides, Spodoptera frugiperda Cad (SfCad) and Manduca sexta Cad (MsCad), which differentially enhance Cry1Fa toxicity to Spodoptera frugiperda neonates. We show that differential SfCad- and MsCad-mediated protection of Cry1Fa toxin in the Spodoptera frugiperda midgut correlates with differential Cry1Fa toxicity enhancement. Both peptides exhibited high affinity for Cry1Fa toxin and an increased rate of Cry1Fa-induced pore formation in S. frugiperda. However, only SfCad bound the S. frugiperda brush border membrane vesicle and more effectively prolonged the stability of Cry1Fa toxin in the gut, explaining higher Cry1Fa enhancement by this peptide. This study shows that cadherin fragments may enhance B. thuringiensis toxicity by at least two different mechanisms or a combination thereof: (i) protection of Cry toxin from protease degradation in the insect midgut and (ii) enhancement of pore-forming ability of Cry toxin. PMID:22081566

  18. Quintuplet Cluster

    NASA Technical Reports Server (NTRS)

    1999-01-01

    Penetrating 25,000 light-years of obscuring dust and myriad stars, NASA's Hubble Space Telescope has provided the clearest view yet of one of the largest young clusters of stars inside our Milky Way galaxy, located less than 100 light-years from the very center of the Galaxy. Having the equivalent mass greater than 10,000 stars like our sun, the monster cluster is ten times larger than typical young star clusters scattered throughout our Milky Way. It is destined to be ripped apart in just a few million years by gravitational tidal forces in the galaxy's core. But in its brief lifetime it shines more brightly than any other star cluster in the Galaxy. Quintuplet Cluster is 4 million years old. It has stars on the verge of blowing up as supernovae. It is the home of the brightest star seen in the galaxy, called the Pistol star. This image was taken in infrared light by Hubble's NICMOS camera in September 1997. The false colors correspond to infrared wavelengths. The galactic center stars are white, the red stars are enshrouded in dust or behind dust, and the blue stars are foreground stars between us and the Milky Way's center. The cluster is hidden from direct view behind black dust clouds in the constellation Sagittarius. If the cluster could be seen from earth it would appear to the naked eye as a 3rd magnitude star, 1/6th of a full moon's diameter apart.

  19. Identification and characterization of FaSOC1, a homolog of SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 from strawberry.

    PubMed

    Lei, Heng-Jiu; Yuan, Hua-Zhao; Liu, Yun; Guo, Xin-Wei; Liao, Xiong; Liu, Lin-Lin; Wang, Qi; Li, Tian-Hong

    2013-12-01

    A MADS-box gene SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1) integrates multiple flowering signals to regulate floral transition in Arabidopsis. Strawberry (Fragaria spp.) is an economically important fruit crop, but its molecular control of flowering is largely unknown. In this study, a SOC1-like gene, FaSOC1, was isolated and characterized from strawberry. The open reading frame of FaSOC1 was 648bp, encoding a protein of 215 amino acids. Sequence alignment and phylogenetic analysis showed that the FaSOC1 protein contained a highly conserved MADS domain and a SOC1 motif, and that it was a member of the SOC1-like genes of dicots. The FaSOC1 protein mainly localized in the cytoplasm of onion epidermal cells and Arabidopsis protoplasts, and showed no transcriptional activation activity in yeast cells. Under the floral induction conditions, the expression of FaSOC1 increased during the first 2weeks of short-day treatment, but declined dramatically during three to 4weeks. FaSOC1 was highly expressed in reproductive organs, including shoot apices, floral buds, flowers, stamens and sepals. Overexpression of FaSOC1 in wild-type Arabidopsis caused early flowering and upregulated the expression of flowering time genes LFY and AP1. In addition, the yeast two-hybrid and BiFC assays confirmed that FaSOC1 could interact with AGL24. In conclusion, these results suggest that FaSOC1 is a flowering promoter in strawberry. PMID:24055423

  20. Glucose transporter levels in tissues of spontaneously diabetic Zucker fa/fa rat (ZDF/drt) and viable yellow mouse (Avy/a).

    PubMed

    Slieker, L J; Sundell, K L; Heath, W F; Osborne, H E; Bue, J; Manetta, J; Sportsman, J R

    1992-02-01

    We used antibodies to the fat/muscle glucose transporter (GLUT4) and the liver glucose transporter (GLUT2) to measure levels of these proteins in various tissues of two rodent models of non-insulin-dependent (type II) diabetes mellitus: the obese spontaneously diabetic male Zucker fa/fa rat (ZDF/drt) and the male viable yellow Avy/a obese diabetic mouse. The ZDF/drt strain generally develops overt diabetes associated with decreased plasma insulin levels. Depending on the age of the animals, the ZDF/drt rats can be arbitrarily segregated into age-matched obese, mildly diabetic (blood glucose less than 11 mM) and obese, and severely diabetic (blood glucose greater than 20 mM) groups. Avy/a mice are comparably hyperglycemic but unlike the ZDF/drt rats are severely hyperinsulinemic. In both groups of diabetic animals, GLUT4 in adipose tissue, heart, and skeletal muscle was reduced 25-55%, and GLUT2 in liver was increased 30-40%, relative to lean, age-matched controls. However, when the mildly diabetic ZDF/drt rats were compared to the lean controls, the only significant difference was a 25% reduction of GLUT4 in heart. Within all of the ZDF/drt rats (excluding the lean controls), GLUT2 in liver and GLUT4 in adipose tissue, heart, and skeletal muscle correlated significantly with glycemia. These data suggest that, in these two models of type II diabetes, glucose transporter levels in muscle, adipose tissue, and liver are regulated in a tissue-selective manner in response to changes in insulin and glucose. Furthermore, at least in the ZDF/drt rat, alterations in GLUT2 and/or GLUT4 protein levels appear not to be associated with obesity per se but appear to be secondary to the severely diabetic state.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1733808

  1. Galaxy Clusters

    NASA Astrophysics Data System (ADS)

    Miller, Christopher J. Miller

    2012-03-01

    There are many examples of clustering in astronomy. Stars in our own galaxy are often seen as being gravitationally bound into tight globular or open clusters. The Solar System's Trojan asteroids cluster at the gravitational Langrangian in front of Jupiter’s orbit. On the largest of scales, we find gravitationally bound clusters of galaxies, the Virgo cluster (in the constellation of Virgo at a distance of ˜50 million light years) being a prime nearby example. The Virgo cluster subtends an angle of nearly 8◦ on the sky and is known to contain over a thousand member galaxies. Galaxy clusters play an important role in our understanding of theUniverse. Clusters exist at peaks in the three-dimensional large-scale matter density field. Their sky (2D) locations are easy to detect in astronomical imaging data and their mean galaxy redshifts (redshift is related to the third spatial dimension: distance) are often better (spectroscopically) and cheaper (photometrically) when compared with the entire galaxy population in large sky surveys. Photometric redshift (z) [Photometric techniques use the broad band filter magnitudes of a galaxy to estimate the redshift. Spectroscopic techniques use the galaxy spectra and emission/absorption line features to measure the redshift] determinations of galaxies within clusters are accurate to better than delta_z = 0.05 [7] and when studied as a cluster population, the central galaxies form a line in color-magnitude space (called the the E/S0 ridgeline and visible in Figure 16.3) that contains galaxies with similar stellar populations [15]. The shape of this E/S0 ridgeline enables astronomers to measure the cluster redshift to within delta_z = 0.01 [23]. The most accurate cluster redshift determinations come from spectroscopy of the member galaxies, where only a fraction of the members need to be spectroscopically observed [25,42] to get an accurate redshift to the whole system. If light traces mass in the Universe, then the locations

  2. Occupational Clusters.

    ERIC Educational Resources Information Center

    Pottawattamie County School System, Council Bluffs, IA.

    The 15 occupational clusters (transportation, fine arts and humanities, communications and media, personal service occupations, construction, hospitality and recreation, health occupations, marine science occupations, consumer and homemaking-related occupations, agribusiness and natural resources, environment, public service, business and office…

  3. Data Clustering

    NASA Astrophysics Data System (ADS)

    Wagstaff, Kiri L.

    2012-03-01

    On obtaining a new data set, the researcher is immediately faced with the challenge of obtaining a high-level understanding from the observations. What does a typical item look like? What are the dominant trends? How many distinct groups are included in the data set, and how is each one characterized? Which observable values are common, and which rarely occur? Which items stand out as anomalies or outliers from the rest of the data? This challenge is exacerbated by the steady growth in data set size [11] as new instruments push into new frontiers of parameter space, via improvements in temporal, spatial, and spectral resolution, or by the desire to "fuse" observations from different modalities and instruments into a larger-picture understanding of the same underlying phenomenon. Data clustering algorithms provide a variety of solutions for this task. They can generate summaries, locate outliers, compress data, identify dense or sparse regions of feature space, and build data models. It is useful to note up front that "clusters" in this context refer to groups of items within some descriptive feature space, not (necessarily) to "galaxy clusters" which are dense regions in physical space. The goal of this chapter is to survey a variety of data clustering methods, with an eye toward their applicability to astronomical data analysis. In addition to improving the individual researcher’s understanding of a given data set, clustering has led directly to scientific advances, such as the discovery of new subclasses of stars [14] and gamma-ray bursts (GRBs) [38]. All clustering algorithms seek to identify groups within a data set that reflect some observed, quantifiable structure. Clustering is traditionally an unsupervised approach to data analysis, in the sense that it operates without any direct guidance about which items should be assigned to which clusters. There has been a recent trend in the clustering literature toward supporting semisupervised or constrained

  4. Cluster generator

    DOEpatents

    Donchev, Todor I.; Petrov, Ivan G.

    2011-05-31

    Described herein is an apparatus and a method for producing atom clusters based on a gas discharge within a hollow cathode. The hollow cathode includes one or more walls. The one or more walls define a sputtering chamber within the hollow cathode and include a material to be sputtered. A hollow anode is positioned at an end of the sputtering chamber, and atom clusters are formed when a gas discharge is generated between the hollow anode and the hollow cathode.

  5. A modified F/A-18 in a distinctive red, white and blue paint scheme was showcased during formal roll

    NASA Technical Reports Server (NTRS)

    2002-01-01

    A modified F/A-18 in a distinctive red, white and blue paint scheme was showcased during formal rollout ceremonies for the Active Aeroelastic Wing flight research program at NASA's Dryden Flight Research Center on March 27, 2002.

  6. Purification and Characterization of Botulinum Neurotoxin FA from a Genetically Modified Clostridium botulinum Strain

    PubMed Central

    Pellett, Sabine; Tepp, William H.; Bradshaw, Marite; Kalb, Suzanne R.; Dykes, Janet K.; Lin, Guangyun; Nawrocki, Erin M.; Pier, Christina L.; Barr, John R.; Maslanka, Susan E.

    2016-01-01

    ABSTRACT Botulinum neurotoxins (BoNTs), produced by neurotoxigenic clostridial species, are the cause of the severe disease botulism in humans and animals. Early research on BoNTs has led to their classification into seven serotypes (serotypes A to G) based upon the selective neutralization of their toxicity in mice by homologous antibodies. Recently, a report of a potential eighth serotype of BoNT, designated “type H,” has been controversial. This novel BoNT was produced together with BoNT/B2 in a dual-toxin-producing Clostridium botulinum strain. The data used to designate this novel toxin as a new serotype were derived from culture supernatant containing both BoNT/B2 and novel toxin and from sequence information, although data from two independent laboratories indicated neutralization by antibodies raised against BoNT/A1, and classification as BoNT/FA was proposed. The sequence data indicate a chimeric structure consisting of a BoNT/A1 receptor binding domain, a BoNT/F5 light-chain domain, and a novel translocation domain most closely related to BoNT/F1. Here, we describe characterization of this toxin purified from the native strain in which expression of the second BoNT (BoNT/B) has been eliminated. Mass spectrometry analysis indicated that the toxin preparation contained only BoNT/FA and confirmed catalytic activity analogous to that of BoNT/F5. The in vivo mouse bioassay indicated a specific activity of this toxin of 3.8 × 107 mouse 50% lethal dose (mLD50) units/mg, whereas activity in cultured human neurons was very high (50% effective concentration [EC50] = 0.02 mLD50/well). Neutralization assays in cells and mice both indicated full neutralization by various antibodies raised against BoNT/A1, although at 16- to 20-fold-lower efficiency than for BoNT/A1. IMPORTANCE Botulinum neurotoxins (BoNTs), produced by anaerobic bacteria, are the cause of the potentially deadly, neuroparalytic disease botulism. BoNTs have been classified into seven serotypes

  7. Purification and Characterization of Botulinum Neurotoxin FA from a Genetically Modified Clostridium botulinum Strain.

    PubMed

    Pellett, Sabine; Tepp, William H; Bradshaw, Marite; Kalb, Suzanne R; Dykes, Janet K; Lin, Guangyun; Nawrocki, Erin M; Pier, Christina L; Barr, John R; Maslanka, Susan E; Johnson, Eric A

    2016-01-01

    Botulinum neurotoxins (BoNTs), produced by neurotoxigenic clostridial species, are the cause of the severe disease botulism in humans and animals. Early research on BoNTs has led to their classification into seven serotypes (serotypes A to G) based upon the selective neutralization of their toxicity in mice by homologous antibodies. Recently, a report of a potential eighth serotype of BoNT, designated "type H," has been controversial. This novel BoNT was produced together with BoNT/B2 in a dual-toxin-producing Clostridium botulinum strain. The data used to designate this novel toxin as a new serotype were derived from culture supernatant containing both BoNT/B2 and novel toxin and from sequence information, although data from two independent laboratories indicated neutralization by antibodies raised against BoNT/A1, and classification as BoNT/FA was proposed. The sequence data indicate a chimeric structure consisting of a BoNT/A1 receptor binding domain, a BoNT/F5 light-chain domain, and a novel translocation domain most closely related to BoNT/F1. Here, we describe characterization of this toxin purified from the native strain in which expression of the second BoNT (BoNT/B) has been eliminated. Mass spectrometry analysis indicated that the toxin preparation contained only BoNT/FA and confirmed catalytic activity analogous to that of BoNT/F5. The in vivo mouse bioassay indicated a specific activity of this toxin of 3.8 × 10(7) mouse 50% lethal dose (mLD50) units/mg, whereas activity in cultured human neurons was very high (50% effective concentration [EC50] = 0.02 mLD50/well). Neutralization assays in cells and mice both indicated full neutralization by various antibodies raised against BoNT/A1, although at 16- to 20-fold-lower efficiency than for BoNT/A1. IMPORTANCE Botulinum neurotoxins (BoNTs), produced by anaerobic bacteria, are the cause of the potentially deadly, neuroparalytic disease botulism. BoNTs have been classified into seven serotypes, serotypes A

  8. Façade mapping and data storage of historical structure ashlars

    NASA Astrophysics Data System (ADS)

    Holzer, R.; Bednarik, M.; Kovarova, K.; Laho, M.; Duncko, M.

    2012-04-01

    The aim of the introduced research is to generate the methodology for the research and maintenance of facing ashlars of historical structures. As a rule, the natural building stone of historic monuments is subject to various types of deterioration and damages mostly due to weathering processes. Preferably, the damaged stone ashlars should be replaced by the natural material of the same lithological composition, of an appropriate durability and the same or similar appearance. If the quarrying of the original stone is not prospective, the quality of alternative rock is moreover not only a crucial condition, but also the fact that its use will not change the façade appearance. The methodology comprehensively summarizes technical, technological and monumental demands. It is oriented on the definition and verification of selection criteria of the most appropriate alternative stone recommended for building or restoration purposes. Before the restoration it is methodologically necessary to generate the ashlars façade plan based on digital photographic procedure (real scale) and data processing in GIS, to assess the lithology of the façade stones and to perform complex laboratory tests (when samples taking is possible) to determine physical-mechanical properties, as UCS, absorption capacity, coefficients of weakening and freezing, etc. aiming at the stone durability. The detailed survey of the replacement building stone has to be methodically identical with the research of facades stone properties. The comparative analysis of the appearance, properties and durability of stone is necessary, as well. Using such methodology it will be possible to design the adequate engineering-historical research and propose the optimal procedure for the restoration of concrete historical structure. Such a comprehensive assessment of original and alternative natural building stone contributes to the protection against undesirable interventions in the restoration of historical monuments.

  9. Water tunnel flow visualisation of vortex breakdown over the F/A-18

    NASA Astrophysics Data System (ADS)

    Thompson, D. H.

    1990-12-01

    Vortex flow patterns over models of the F/A-18 aircraft were visualized using dye and hydrogen bubble techniques in a water tunnel. The axial position of vortex breakdown in the leading-edge extension (LEX) vortices was measured, and was found to be insensitive to Reynolds number, to flap setting, and to small variations in model cross-section shape. Engine inlet flow did alter the vortex breakdown position, at flow rates that might be encountered under flight conditions. The fitting of fences on the LEX upper surface did not affect the axial position of vortex breakdown, but did alter the vortex structure. These alterations were examined in some detail.

  10. Myosins FaMyo2B and Famyo2 Affect Asexual and Sexual Development, Reduces Pathogenicity, and FaMyo2B Acts Jointly with the Myosin Passenger Protein FaSmy1 to Affect Resistance to Phenamacril in Fusarium asiaticum.

    PubMed

    Zheng, Zhitian; Liu, Xiumei; Li, Bin; Cai, Yiqiang; Zhu, Yuanye; Zhou, Mingguo

    2016-01-01

    We previously reported that mutations occurred in the gene myosin5 were responsible for resistance to the fungicide phenamacril in Fusarium graminearum. Here, we determined whether there is a functional link between phenamacril resistance and the myosin proteins FaMyo2B and Famyo2 in Fusarium asiaticum, which is the major causal agent of Fusarium head blight in China. We found that FaMyo2B acts jointly with FaSmy1 to affect resistance to phenamacril in F. asiaticum. We also found that FaMyo2B disruption mutant and Famyo2 deletion mutant were defective in hyphal branching, conidiation, and sexual reproduction. ΔFamyo2 also had an enhanced sensitivity to cell wall damaging agents and an abnormal distribution of septa and nuclei. In addition, the FaMyo2B and Famyo2 mutants had reduced pathogenicity on wheat coleoptiles and flowering wheat heads. Taken together, these results reveal that FaMyo2B and Famyo2 are required for several F. asiaticum developmental processes and activities, which help us better understand the resistance mechanism and find the most effective approach to control FHB. PMID:27099966

  11. Myosins FaMyo2B and Famyo2 Affect Asexual and Sexual Development, Reduces Pathogenicity, and FaMyo2B Acts Jointly with the Myosin Passenger Protein FaSmy1 to Affect Resistance to Phenamacril in Fusarium asiaticum

    PubMed Central

    Zheng, Zhitian; Liu, Xiumei; Li, Bin; Cai, Yiqiang; Zhu, Yuanye; Zhou, Mingguo

    2016-01-01

    We previously reported that mutations occurred in the gene myosin5 were responsible for resistance to the fungicide phenamacril in Fusarium graminearum. Here, we determined whether there is a functional link between phenamacril resistance and the myosin proteins FaMyo2B and Famyo2 in Fusarium asiaticum, which is the major causal agent of Fusarium head blight in China. We found that FaMyo2B acts jointly with FaSmy1 to affect resistance to phenamacril in F. asiaticum. We also found that FaMyo2B disruption mutant and Famyo2 deletion mutant were defective in hyphal branching, conidiation, and sexual reproduction. ΔFamyo2 also had an enhanced sensitivity to cell wall damaging agents and an abnormal distribution of septa and nuclei. In addition, the FaMyo2B and Famyo2 mutants had reduced pathogenicity on wheat coleoptiles and flowering wheat heads. Taken together, these results reveal that FaMyo2B and Famyo2 are required for several F. asiaticum developmental processes and activities, which help us better understand the resistance mechanism and find the most effective approach to control FHB. PMID:27099966

  12. Positional distribution of FA in TAG of enzymatically modified borage and evening primrose oils.

    PubMed

    Senanayake, S P J Namal; Shahidi, Fereidoon

    2002-08-01

    Stereospecific analysis was carried out to establish positional distribution of FA in the TAG of DHA, EPA, and (EPA + DHA)-enriched oils. In this study, TAG of enzymatically modified oils were purified using a silicic acid column. The TAG were then subjected to positional distribution analysis using a modified procedure involving reductive cleavage with Grignard reagent. The results showed that in DHA-enriched borage oil (BO), DHA was randomly distributed over the three positions of TAG, whereas gamma-linolenic acid (GLA) was mainly esterified at the sn-2 and -3 positions. In DHA-enriched evening primrose oil (EPO), however, DHA and GLA were concentrated in the sn-2 position. In EPA-enriched BO, EPA was randomly distributed over the three positions of TAG, similar to that observed for DHA. In EPA-enriched EPO, however, this FA was mainly located at the primary positions (sn-1 and sn-3) of TAG. In both oils, GLA was preferentially esterified at the sn-2 position. In (EPA + DHA)-enriched BO, EPA and DHA were mainly esterified at the sn-1 and -3 positions of TAG, whereas GLA was mainly located at the sn-2 position. In (EPA + DHA)-enriched EPO, GLA was mainly located at the sn-2 and -3 positions; EPA was preferentially esterified at the sn-1 and -3 positions, and DHA was found mainly at the sn-3 position. PMID:12371752

  13. Overview of HATP Experimental Aerodynamics Data for the Baseline F/A-18 Configuration

    NASA Technical Reports Server (NTRS)

    Hall, Robert M.; Murri, Daniel G.; Erickson, Gary E.; Fisher, David F.; Banks, Daniel W.; Lanser, Wendy, R.

    1996-01-01

    Determining the baseline aerodynamics of the F/A-18 was one of the major objectives of the High-Angle-of-Attack Technology Program (HATP). This paper will review the key data bases that have contributed to our knowledge of the baseline aerodynamics and the improvements in test techniques that have resulted from the experimental program. Photographs are given highlighting the forebody and leading-edge-extension (LEX) vortices. Other data representing the impact of Mach and Reynolds numbers on the forebody and LEX vortices will also be detailed. The level of agreement between different tunnels and between tunnels and flight will be illustrated using pressures, forces, and moments measured on a 0.06-scale model tested in the Langley 7- by 10-Foot High Speed Tunnel, a 0.16-scale model in the Langley 30- by 60-Foot Tunnel, a full-scale vehicle in the Ames 80- by 120-Foot Wind Tunnel, and the flight F/A-18 High Alpha Research Vehicle (HARV). Next, creative use of wind tunnel resources that accelerated the validation of the computational fluid dynamics (CFD) codes will be described. Lastly, lessons learned, deliverables, and program conclusions are presented.

  14. Aerostructural Vortical Flow Interactions with Applications to F/A-18 and F-117 Tail Buffet

    NASA Technical Reports Server (NTRS)

    Kandil, Osama A.; Massey, Steven J.; Sheta, Essam F.

    1996-01-01

    The buffet response of the flexible twin-tail configuration of the F/A-18 and a generic F-111 aircraft are computationally simulated and experimentally validated. The problem is a multidisciplinary one which requires the sequential solution of three sets of equations on a multi-block grid structure. The first set is the unsteady, compressible, full Navier-Stokes equations. The second set is the aeroelastic equations for bending and torsional twin-tail responses. The third set is the grid-displacement equations which are used to update the grid coordinates due to the tail deflections. The computational models consist of a 76 deg. swept back, sharp edged delta wing of aspect ratio of one and a swept-back F/A-18 or F-117 twin-tail. The configuration is pitched at 30 deg. angle-of-attack. The problem is solved for the initial flow conditions with the twin tails kept rigid. Next, the aeroelastic equations of the tails are turned on along with the grid-displacement equations to solve for the bending and torsional tails responses due to the unsteady loads produced by the vortex breakdown flow of the leading-edge vortex cores of the delta wing. Several spanwise locations of the twin tails are investigated. The computational results are validated using several existing experimental data.

  15. Offspring Production among the Extended Relatives of Samoan Men and Fa'afafine

    PubMed Central

    VanderLaan, Doug P.; Forrester, Deanna L.; Petterson, Lanna J.; Vasey, Paul L.

    2012-01-01

    Androphilia refers to sexual attraction to adult males, whereas gynephilia refers to sexual attraction to adult females. Male androphilia is an evolutionary paradox. Its development is at least partially influenced by genetic factors, yet male androphiles exhibit lower reproductive output, thus raising the question of how genetic factors underlying its development persist. The sexual antagonism hypothesis posits that the fitness costs associated with genetic factors underlying male androphilia are offset because these same factors lead to elevated reproduction on the part of the female relatives of androphilic males. Western samples drawn from low fertility populations have yielded inconsistent results when testing this hypothesis. Some studies documented elevated reproduction among the matrilineal female kin of androphilic males, whereas others found such effects in the paternal line. Samoa is a high-fertility population in which individuals reproduce closer to their maximum capacities. This study compared the reproductive output of the paternal and maternal line grandmothers, aunts, and uncles of 86 Samoan androphilic males, known locally as fa'afafine, and 86 Samoan gynephilic males. Reproductive output was elevated in the paternal and maternal line grandmothers, but not aunts or uncles, of fa'afafine. These findings are consistent with the sexual antagonism hypothesis and suggest that male androphilia is associated with elevated reproduction among extended relatives in both the maternal and paternal line. Discussion focuses on how this study, in conjunction with the broader literature, informs various models for the evolution of male androphilia via elevated reproduction on the part of female kin. PMID:22558342

  16. Composting of food waste subjected to hydrothermal pretreatment and inoculated with Paecilomyces sp. FA13.

    PubMed

    Nakasaki, Kiyohiko; Mimoto, Hiroshi; Tran, Quyen Ngoc Minh; Oinuma, Akiko

    2015-03-01

    Food waste collected from restaurants, convenience stores, and food-processing factories was mixed with sawdust and subjected to hydrothermal pretreatment at 180°C for 30min to prepare compost raw material. Furan compounds such as 5-HMF (5-hydroxymethyl furfural) and furfural were produced at concentration levels of approximately 8 and 0.5mg/g-ds, respectively, through hydrothermal pretreatment. The furan compounds inhibited the activity of composting microorganisms, thus delaying the start of organic matter degradation during composting. A newly identified fungus, Paecilomyces sp. FA13, which possesses the ability to degrade furan compounds, was isolated and used as an inoculum for the composting of the raw material prepared by hydrothermal pretreatment. By inoculating the FA13 into the compost raw material at 10(5)CFU/g-ds, the degradation of furan compounds was accelerated. As a result, bacterial activity, which contributed to composting, was enhanced, significantly promoting the start of vigorous degradation of organic materials. PMID:25585259

  17. F/A-18 Performance Benefits Measured During the Autonomous Formation Flight Project

    NASA Technical Reports Server (NTRS)

    Vachon, M. Jake; Ray, Ronald J.; Walsh, Kevin R.; Ennix, Kimberly

    2003-01-01

    The Autonomous Formation Flight (AFF) project at the NASA Dryden Flight Research Center (Edwards, California) investigated performance benefits resulting from formation flight, such as reduced aerodynamic drag and fuel consumption. To obtain data on performance benefits, a trailing F/A-18 airplane flew within the wing tip-shed vortex of a leading F/A-18 airplane. The pilot of the trail airplane used advanced station-keeping technology to aid in positioning the trail airplane at precise locations behind the lead airplane. The specially instrumented trail airplane was able to obtain accurate fuel flow measurements and to calculate engine thrust and vehicle drag. A maneuver technique developed for this test provided a direct comparison of performance values while flying in and out of the vortex. Based on performance within the vortex as a function of changes in vertical, lateral, and longitudinal positioning, these tests explored design-drivers for autonomous stationkeeping control systems. Observations showed significant performance improvements over a large range of trail positions tested. Calculations revealed maximum drag reductions of over 20 percent, and demonstrated maximum reductions in fuel flow of just over 18 percent.

  18. Occultifur tropicalis f.a., sp. nov., a novel cystobasidiomycetous yeast species isolated from tropical regions.

    PubMed

    Khunnamwong, Pannida; Surussawadee, Janjira; Jindamorakot, Sasitorn; Ribeiro, José R A; Hagler, Allen N; Limtong, Savitree

    2015-05-01

    Five strains representing a single novel anamorphic yeast species were isolated from sugar cane. Two strains were from tissue (DMKU-SE38, DMKU-SE59(T)) and two from the external surface (DMKU-SP385, DMKU-SP403) of leaves collected in Thailand and the fifth (IMUFRJ 52020) from the rhizoplane of sugar cane in an organically cultivated field in Brazil. On the basis of sequence analysis of the D1/D2 region of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) region, they were classified as representing a single species of the genus Occultifur. The sequences of the D1/D2 region of the LSU rRNA genes and the ITS regions of the five strains were either identical or differed from each other by only one nucleotide substitution. The novel species was related most closely to Occultifur brasiliensis f.a. CBS 12687(T) but with 0.7-1.0% nucleotide substitutions (4-6 nt) in the D1/D2 region of the LSU rRNA gene and 2.5-2.7% nucleotide substitutions (14-15 nt) in the ITS region. The name Occultifur tropicalis f.a., sp. nov. is proposed. The type strain is DMKU-SE59(T) ( =BCC 61184(T) =NBRC 109696(T) =CBS 13389 (T)). PMID:25713048

  19. F/A-18 forebody vortex control. Volume 1: Static tests

    NASA Technical Reports Server (NTRS)

    Kramer, Brian R.; Suarez, Carlos J.; Malcolm, Gerald N.; Ayers, Bert F.

    1994-01-01

    A wind tunnel test was conducted on a six percent model of the F/A-18 at the NASA Ames 7 X 10-Foot Low Speed Wind Tunnel. The primary objective of the test was to evaluate several forebody vortex control configurations at high angles of attack in order to determine the most effective method of obtaining well behaved yawing moments, in preparation for the rotary balance test. Both mechanical and pneumatic systems were tested. Single and dual rotating nose tip strakes and a vertical nose strake were tested at different sizes and deflections. A series of jet blowing configurations were located at various fuselage stations, azimuth angles, and pointing angles ranging from straight aft to 60 deg canted inboard. Slot blowing was investigated for several slot lengths and fuselage stations. The effect of blowing rate was tested for both of these pneumatic systems. The most effective configurations were then further tested with a variation of both sideslip angle and Reynolds number over a range of angles of attack from 0 to 60 deg. It was found that a very robust system can be developed that provides yawing moments at angles of attack up to 60 deg that significantly exceeds that available from 30 deg of rudder deflection (F/A-18 maximum) at 0 deg angle of attack.

  20. Stability analysis of an F/A-18 E/F cable mount m odel

    NASA Technical Reports Server (NTRS)

    Thompson, Nancy; Farmer, Moses

    1994-01-01

    A full-span F/A-18 E/F cable mounted wind tunnel model is part of a flutter clearance program at the NASA Langley Transonic Dynamics Tunnel. Parametric analysis of this model using GRUMCBL software was conducted to assess stability for wind tunnel tests. Two configurations of the F/A-18 E/F were examined. The parameters examined were pulley-cable friction, mach number, dynamic pressure, cable geometry, center of gravity location, cable tension, snubbing the model, drag, and test medium. For the nominal cable geometry (Cable Geometry 1), Configuration One was unstable for cases with higher pulley-cable friction coefficients. A new cable geometry (Cable Geometry 3) was determined in which Configuration One was stable for all cases evaluated. Configuration Two with the nominal center of gravity position was found to be unstable for cases with higher pulley-cable friction coefficients; however, the model was stable when the center of gravity moved forward 1/2. The model was tested using the cable mount system during the initial wind tunnel entry and was stable as predicted.

  1. Cytotoxicity of the dicarboximide fungicides, vinclozolin and iprodione, in rat hepatoma-derived Fa32 cells.

    PubMed

    Dierickx, Paul J

    2004-10-01

    Dicarboximide fungicides are widely used to control various fungal species. Their primary action is not known, due to a lack of knowledge concerning the mechanism of action of the dicarboximide group. The cytotoxicities of vinclozolin and iprodione in rat hepatoma-derived Fa32 cells were investigated. Cytotoxicity was measured by neutral red uptake inhibition after treatment for 24 hours. Iprodione was more toxic than vinclozolin. Vinclozolin was less toxic in glutathione-depleted cells than in control cells. This was also true for iprodione at lower concentrations, but iprodione became more toxic at higher concentrations. Both the fungicides increased the endogenous glutathione content by 20% after 1 hour. After 24 hours, the glutathione content was doubled by vinclozolin, but was not affected by iprodione. No effect on glutathione S-transferase activity or reactive oxygen species formation could be observed. Cytochrome P450-dependent ethoxyresorufin-O-deethylase and pentoxyresorufin-O-depentylase activities were moderately activated by iprodione and strongly activated by vinclozolin. A glutathione-related cytochrome P450-dependent metabolic attack of vinclozolin and iprodione could be responsible for their cytotoxicity in Fa32 cells. Further research is needed to fully elucidate these (or other) mechanisms. PMID:15651921

  2. Cytotoxicity of the MEIC reference chemicals in rat hepatoma-derived Fa32 cells.

    PubMed

    Dierickx, P J

    2000-09-01

    The cytotoxicity of the MEIC reference chemicals was investigated in rat hepatoma-derived Fa32 cells. The total protein content was measured as an endpoint after exposure times of 30 min and 24 h, both in normal and glutathione-depleted cells. The neutral red uptake inhibition and the MTT conversion were also measured after 30 min. On average, the cytotoxicity was higher in glutathione-depleted cells when compared to normal cells, and was lower after 30 min than after 24 h. Evidence was obtained for lysosomal attack (of five chemicals) or mitochondrial dysfunction (of six chemicals) as the primary intoxication mechanism. Malathion and mercuric chloride belong to both series of chemicals. Good to excellent correlations were observed when the 50% inhibitory concentrations of the six different in vitro assays were compared. When the six in vitro assays in Fa32 cells were compared with the human toxicity, the correlation coefficient was almost always identical to that obtained previously in human hepatoma-derived Hep G2 cells. The latter was the best acute in vitro assay for the prediction of human toxicity within the MEIC study. Altogether the results integrate very well with the basal cytotoxicity concept (Ekwall, B., 1995. The basal cytotoxicity concept. In: Goldberg, A.M., Van Zutphen, L.F.M. (Eds.), The World Congress on Alternatives and Animal Use in the Life Sciences: Education, Research, Testing. Mary Ann Liebert Publishers, New York, pp. 721-725). PMID:10996672

  3. Noise cancellation for CELP voice encoders in an F/A-18 noise environment

    NASA Astrophysics Data System (ADS)

    Heide, David A.

    1994-03-01

    Because of the severe noise environment in the Navy's F/A-18 jet aircraft, it has always been very difficult to achieve highly intelligible speech using low data rate voice encoders such as the 2.4 kbps LPC-10. As a result, all voice encoding has been done with a high data rate 16.0 kbps CVSD algorithm. The main focus of this research was to develop a technique that could retain the acceptable intelligibility of the high rate encoders while still significantly lowering the data rate required. To achieve these results, a noise cancellation preprocessor was developed to be used in tandem with the new 4.8 kbps CELP encoder that is being implemented in the STU-3. While many noise reduction techniques were investigated, the best results were accomplished by first reducing the noise through spectral subtraction and then enhancing the important resonant formants of the speech. The results indicated that when the noise cancellation preprocessor was added to the CELP encoder, the DRT intelligibility scores were improved by a significant 5.0 points, making the speech much more acceptable for possible use in the F/A-18.

  4. In vitro effects of fat, FA, and cholesterol on sphingomyelin hydrolysis induced by rat intestinal alkaline sphingomyelinase.

    PubMed

    Liu, Jian-Jun; Nilsson, Ake; Duan, Rui-Dong

    2002-05-01

    Dietary sphingomyelin (SM) may have regulatory effects on cell proliferation and tumorigenesis in the colon. Alkaline sphingomyelinase (SMase) is the major enzyme responsible for hydrolysis of SM in the gut. Previously we purified the enzyme and showed that the presence of glycerophospholipids inhibited SM hydrolysis induced by alkaline SMase in vitro. In the present work, we studied the effects of TG, DG, FA, ceramide, and cholesterol on SM hydrolysis catalyzed by purified alkaline SMase. The results showed that both TG (triolein and tristearin) and DG (1,2-dioleoyl-sn-glycerol and 1,2-distearoyl-rac-glycerol) inhibited the activity of alkaline SMase. 1-Monooleoyl-rac-glycerol, 1-monostearoyl-rac-glycerol, stearic acid, oleic acid, linoleic acid, linolenic acid, and arachidonic acid stimulated the activity of alkaline SMase at 0.4-0.8 mM concentrations but inhibited the enzyme at higher concentrations. There was no difference between the effects induced by saturated and unsaturated FA. A short-chain FA such as lauric acid had a stronger stimulatory effect at low concentrations and weaker inhibitory effect at high concentrations than long-chain FA. Choosing linoleic acid as an example, we found that FA had similar effects on both alkaline SMase and neutral SMase. Cholesterol and ceramide when mixed with FA to increase its solubility in bile salt micelles inhibited SMase activity. In conclusion, glycerides, FA, ceramide, and cholesterol influence SM hydrolysis catalyzed by intestinal alkaline SMase. The presence of lipids in the diet may thus influence the course of SM digestion in the gut and thereby the exposure of colon to SM metabolites. PMID:12056588

  5. SVM clustering

    PubMed Central

    Winters-Hilt, Stephen; Merat, Sam

    2007-01-01

    Background Support Vector Machines (SVMs) provide a powerful method for classification (supervised learning). Use of SVMs for clustering (unsupervised learning) is now being considered in a number of different ways. Results An SVM-based clustering algorithm is introduced that clusters data with no a priori knowledge of input classes. The algorithm initializes by first running a binary SVM classifier against a data set with each vector in the set randomly labelled, this is repeated until an initial convergence occurs. Once this initialization step is complete, the SVM confidence parameters for classification on each of the training instances can be accessed. The lowest confidence data (e.g., the worst of the mislabelled data) then has its' labels switched to the other class label. The SVM is then re-run on the data set (with partly re-labelled data) and is guaranteed to converge in this situation since it converged previously, and now it has fewer data points to carry with mislabelling penalties. This approach appears to limit exposure to the local minima traps that can occur with other approaches. Thus, the algorithm then improves on its weakly convergent result by SVM re-training after each re-labeling on the worst of the misclassified vectors – i.e., those feature vectors with confidence factor values beyond some threshold. The repetition of the above process improves the accuracy, here a measure of separability, until there are no misclassifications. Variations on this type of clustering approach are shown. Conclusion Non-parametric SVM-based clustering methods may allow for much improved performance over parametric approaches, particularly if they can be designed to inherit the strengths of their supervised SVM counterparts. PMID:18047717

  6. Adaptive Augmenting Control Flight Characterization Experiment on an F/A-18

    NASA Technical Reports Server (NTRS)

    VanZwieten, Tannen S.; Orr, Jeb S.; Wall, John H.; Gilligan, Eric T.

    2014-01-01

    This paper summarizes the Adaptive Augmenting Control (AAC) flight characterization experiments performed using an F/A-18 (TN 853). AAC was designed and developed specifically for launch vehicles, and is currently part of the baseline autopilot design for NASA's Space Launch System (SLS). The scope covered here includes a brief overview of the algorithm (covered in more detail elsewhere), motivation and benefits of flight testing, top-level SLS flight test objectives, applicability of the F/A-18 as a platform for testing a launch vehicle control design, test cases designed to fully vet the AAC algorithm, flight test results, and conclusions regarding the functionality of AAC. The AAC algorithm developed at Marshall Space Flight Center is a forward loop gain multiplicative adaptive algorithm that modifies the total attitude control system gain in response to sensed model errors or undesirable parasitic mode resonances. The AAC algorithm provides the capability to improve or decrease performance by balancing attitude tracking with the mitigation of parasitic dynamics, such as control-structure interaction or servo-actuator limit cycles. In the case of the latter, if unmodeled or mismodeled parasitic dynamics are present that would otherwise result in a closed-loop instability or near instability, the adaptive controller decreases the total loop gain to reduce the interaction between these dynamics and the controller. This is in contrast to traditional adaptive control logic, which focuses on improving performance by increasing gain. The computationally simple AAC attitude control algorithm has stability properties that are reconcilable in the context of classical frequency-domain criteria (i.e., gain and phase margin). The algorithm assumes that the baseline attitude control design is well-tuned for a nominal trajectory and is designed to adapt only when necessary. Furthermore, the adaptation is attracted to the nominal design and adapts only on an as-needed basis

  7. t-Bu2SiF-derivatized D2-receptor ligands: the first SiFA-containing small molecule radiotracers for target-specific PET-imaging.

    PubMed

    Iovkova-Berends, Ljuba; Wängler, Carmen; Zöller, Thomas; Höfner, Georg; Wanner, Klaus Theodor; Rensch, Christian; Bartenstein, Peter; Kostikov, Alexey; Schirrmacher, Ralf; Jurkschat, Klaus; Wängler, Björn

    2011-01-01

    The synthesis, radiolabeling and in vitro evaluation of new silicon-fluoride acceptor (SiFA) derivatized D(2)-receptor ligands is reported. The SiFA-technology simplifies the introduction of fluorine-18 into target specific biomolecules for Positron-Emission-Tomography (PET). However, one of the remaining challenges, especially for small molecules such as receptor-ligands, is the bulkiness of the SiFA-moiety. We therefore synthesized four Fallypride SiFA-conjugates derivatized either directly at the benzoic acid ring system (SiFA-DMFP, SiFA-FP, SiFA-DDMFP) or at the butyl-side chain (SiFA-M-FP) and tested their receptor affinities. We found D(2)-receptor affinities for all compounds in the nanomolar range (K(i(SiFA-DMFP)) = 13.6 nM, K(i(SiFA-FP)) = 33.0 nM, K(i(SiFA-DDMFP)) = 62.7 nM and K(i(SiFA-M-FP)) = 4.21 nM). The radiofluorination showed highest yields when 10 nmol of the precursors were reacted with [(18)F]fluoride/TBAHCO(3) in acetonitrile. After a reversed phased cartridge purification the desired products could be isolated as an injectable solution after only 10 min synthesis time with radiochemical yields (RCY) of more than 40% in the case of SiFA-DMFP resulting in specific activities >41 GBq/µmol (>1,100 Ci/mmol). Furthermore, the radiolabeled products were shown to be stable in the injectable solutions, as well as in human plasma, for at least 90 min. PMID:21892125

  8. Interspecific hybridization does not affect the level of fluctuating asymmetry (FA) in the Drosophila bipectinata species complex.

    PubMed

    Banerjee, Parul; Singh, B N

    2015-08-01

    The Drosophila bipectinata species complex comprises of four very closely related species namely D. bipectinata, D. parabipectinata, D. malerkotliana and D. pseudoananassae. It was found that irrespective of the evolutionary divergence among the species, FA which is reflective of the developmental precision remains nearly same in four species. During the present study, the level of FA in different morphological traits was studied in interspecific hybrids and compared with that of parental species with the view that it would throw light on the degree of divergence between the parental species. If they have not diverged much, the interspecific hybrids may have a similar FA level, incompatibilities between their genomes being negligible. On the other hand, if there is substantial divergence, the level of FA may be higher due to incompatibility between the genomes of the parental species. The morphological traits taken were sternopleural bristle number and wing length in both males and females and ovariole number and sex-comb tooth number in females and males respectively. However, except in a few cases, we could not detect any significant differences in the level of FA in hybrids as compared to pure species. On the other hand, a number of abnormalities like poor viability, dystrophied ovaries, asymmetrical eyes etc., could be detected in hybrids from crosses involving D. pseudoananassae as one of the parents. Therefore, we conclude that specific developmental pathways are more susceptible to developmental disturbances due to genomic incompatibilities than the large complex system bringing about developmental stability. PMID:25987248

  9. The fruit ripening-related gene FaAAT2 encodes an acyl transferase involved in strawberry aroma biogenesis.

    PubMed

    Cumplido-Laso, Guadalupe; Medina-Puche, Laura; Moyano, Enriqueta; Hoffmann, Thomas; Sinz, Quirin; Ring, Ludwig; Studart-Wittkowski, Claudia; Caballero, José Luis; Schwab, Wilfried; Muñoz-Blanco, Juan; Blanco-Portales, Rosario

    2012-06-01

    Short-chain esters contribute to the blend of volatiles that define the strawberry aroma. The last step in their biosynthesis involves an alcohol acyltransferase that catalyses the esterification of an acyl moiety of acyl-CoA with an alcohol. This study identified a novel strawberry alcohol acyltransferase gene (FaAAT2) whose expression pattern during fruit receptacle growth and ripening is in accordance with the production of esters throughout strawberry fruit ripening. The full-length FaAAT2 cDNA was cloned and expressed in Escherichia coli and its activity was analysed with acyl-CoA and alcohol substrates. The semi-purified FaAAT2 enzyme had activity with C1-C8 straight-chain alcohols and aromatic alcohols in the presence of acetyl-CoA. Cinnamyl alcohol was the most efficient acyl acceptor. When FaAAT2 expression was transiently downregulated in the fruit receptacle by agroinfiltration, the volatile ester production was significantly reduced in strawberry fruit. The results suggest that FaAAT2 plays a significant role in the production of esters that contribute to the final strawberry fruit flavour. PMID:22563120

  10. The Relationship between Adult Occupational Preferences and Childhood Gender Nonconformity among Samoan Women, Men, and Fa'afafine.

    PubMed

    Semenyna, Scott W; Vasey, Paul L

    2016-09-01

    Previous research has found that sex differences in occupational preferences are both substantial and cross-culturally universal. Androphilic males tend to display "gender-shifted" occupational preferences, with relatively female-typical interests. Past research has overwhelmingly relied on Western samples; this article offers new insights from a non-Western setting. Known locally as fa'afafine, androphilic males in Samoa occupy a third-gender category. Data were collected in Samoa from 103 men, 103 women, and 103 fa'afafine regarding occupational preferences and recalled childhood gender nonconformity (CGN). A substantial sex difference was observed in the occupational preferences of men and women (d = 2.04). Interestingly, women and fa'afafine did not differ in their preferences (p = 0.89), indicating a complete gender inversion of occupational preferences in the latter. Although there was no correlation between women's CGN and masculine occupational preferences, there was a significant correlation (r = -0.62) between these variables in both men and fa'afafine. Among males (both men and fa'afafine), increased CGN was associated with preference for feminine occupations. The present research corroborates past findings and furnishes support for the conclusion that female-typical occupational preferences are a cross-culturally invariant aspect of male androphilia. PMID:27100108

  11. The catalytic kinetic method for the determination of trace formaldehyde (FA) base on a bromate-eosin Y system.

    PubMed

    Tang, Yufang; Chen, Hao; Weng, Chao; Tang, Xiaohui; Zhang, Miaoling; Yang, Qiongqiong; Hu, Tao; Cai, Changqun

    2014-05-01

    A new simple and highly sensitive catalytic kinetic method for the determination of trace amount of FA in food sample has been established. The method was based on the catalytic effect of FA on the oxidation of eosin Y by potassium bromate in present of phosphoric acid. The reaction was monitored spectrophotometrically by measuring the decrease in absorbance of eosin Y at 518 nm. Under the optimized experimental conditions, the developed method allowed the determination of FA in the range of 0.03-0.6 μg mL(-1) with a good precision, and the limit of detection was down to 0.00988 μg mL(-1). The relative standard deviation of five replicate measurements for the determination of FA in concentration 0.12 μg mL(-1) was 1.8%. The proposed method was successfully applied to the determination of FA in food directly and satisfactory results were obtained. PMID:24531542

  12. Transonic Experimental Observations of Abrupt Wing Stall on an F/A-18E Model (Invited)

    NASA Technical Reports Server (NTRS)

    McMillin, S. Naomi; Hall, Robert M.; Lamar, John E.

    2003-01-01

    A transonic wind tunnel test of an 8% F/A-18E model was conducted in the NASA Langley Research Center (LaRC) 16 ft Transonic Tunnel (16-ft TT) to investigate on-surface flow physics during stall. The technical approach employed focused on correlating static (or time-averaged) and unsteady wind-tunnel test data to the unsteady wing-stall events using force, moment, pressure, and pressure-sensitive-paint measurements. This paper focuses on data obtained on the pre-production configuration of the F/A-18E aircraft at Mach number of 0.90. The flow unsteadiness occurring on the wing as the wing went through the stall process was captured using the time history of balance and pressure measurements and by calculating the root mean square (RMS) for a number of instrument signals. The second step was to gather global perspectives on the pressures influencing the wing stall process. The abrupt wing stall experienced by the 8% F/A-18E Model was observed to be an unsteady event triggered by the rapid advancement of separation, which had migrated forward from the trailing edge, to the leading-edge flap hingeline over a very small increment in angle of attack. The angle of attack at which this stall occurred varied, from run to run, over an 1 deg increment. The abrupt wing stall was observed, using pressure-sensitive-paint, to occur simultaneously on both wing panels or asymmetrically. The pressure-sensitive paint data and wingroot bending moment data were essential in providing insight to the flow structures occurring over the wing and the possible asymmetry of those flow structures. A repeatability analysis conducted on eight runs of static data provided a quick and inexpensive examination of the unsteady aerodynamic characteristics of abrupt wing stall. The results of the repeatability analysis agreed extremely well with data obtained using unsteady measurement techniques. This approach could be used to identify test conditions for more complex unsteady data measurements using

  13. Field-Evolved Mode 1 Resistance of the Fall Armyworm to Transgenic Cry1Fa-Expressing Corn Associated with Reduced Cry1Fa Toxin Binding and Midgut Alkaline Phosphatase Expression.

    PubMed

    Jakka, Siva R K; Gong, Liang; Hasler, James; Banerjee, Rahul; Sheets, Joel J; Narva, Kenneth; Blanco, Carlos A; Jurat-Fuentes, Juan L

    2016-02-01

    Insecticidal protein genes from the bacterium Bacillus thuringiensis (Bt) are expressed by transgenic Bt crops (Bt crops) for effective and environmentally safe pest control. The development of resistance to these insecticidal proteins is considered the most serious threat to the sustainability of Bt crops. Resistance in fall armyworm (Spodoptera frugiperda) populations from Puerto Rico to transgenic corn producing the Cry1Fa insecticidal protein resulted, for the first time in the United States, in practical resistance, and Bt corn was withdrawn from the local market. In this study, we used a field-collected Cry1Fa corn-resistant strain (456) of S. frugiperda to identify the mechanism responsible for field-evolved resistance. Binding assays detected reduced Cry1Fa, Cry1Ab, and Cry1Ac but not Cry1Ca toxin binding to midgut brush border membrane vesicles (BBMV) from the larvae of strain 456 compared to that from the larvae of a susceptible (Ben) strain. This binding phenotype is descriptive of the mode 1 type of resistance to Bt toxins. A comparison of the transcript levels for putative Cry1 toxin receptor genes identified a significant downregulation (>90%) of a membrane-bound alkaline phosphatase (ALP), which translated to reduced ALP protein levels and a 75% reduction in ALP activity in BBMV from 456 compared to that of Ben larvae. We cloned and heterologously expressed this ALP from susceptible S. frugiperda larvae and demonstrated that it specifically binds with Cry1Fa toxin. This study provides a thorough mechanistic description of field-evolved resistance to a transgenic Bt crop and supports an association between resistance and reduced Cry1Fa toxin binding and levels of a putative Cry1Fa toxin receptor, ALP, in the midguts of S. frugiperda larvae. PMID:26637593

  14. F/A-18 and F-16 forebody vortex control, static and rotary-balance results

    NASA Technical Reports Server (NTRS)

    Kramer, Brian; Smith, Brooke

    1994-01-01

    The results from research on forebody vortex control on both the F/A-18 and the F-16 aircraft will be shown. Several methods of forebody vortex control, including mechanical and pneumatic schemes, will be discussed. The wind tunnel data includes both static and rotary balance data for forebody vortex control. Time lags between activation or deactivation of the pneumatic control and when the aircraft experiences the resultant forces are also discussed. The static (non-rotating) forces and pressures are then compared to similar configurations tested in the NASA Langley and DTRC Wind Tunnel, the NASA Ames 80'x120' Wind Tunnel, and in flight on the High Angle of Attack Research Vehicle (HARV).

  15. F/A-18 E/F flutter clearance model in the Langley TDT

    NASA Technical Reports Server (NTRS)

    1994-01-01

    An 18 percent aeroelastically-scaled, full span F/A-18 E/F model was tested during multiple wind-tunnel entries in the Langley Transonic Dynamics Tunnel. The primary purpose of these entries was to assist in clearing the flight vehicle design of flutter within its operating envelope. The wind-tunnel model was tested on a string and on a cable-mount system (as shown). All lifting surfaces were flutter cleared up to M=1.2 with the model string mounted. The model was then flutter cleared on the cable-mount system to assess the influence of rigid-body dynamics and fuselage flexibility on flutter. Several configuration parametric studies were also completed, including many external store configurations.

  16. F/A-18 E/F flutter clearance model in the Langley TDT

    NASA Technical Reports Server (NTRS)

    1994-01-01

    An 18 percent aeroelastically-scaled, full span F/A-18 E/F model was tested during multiple wind-tunnel entries in the Langley Transonic Dynamics Tunnel. The primary purpose of these entries was to assist in clearing the flight vehicle design of flutter within its operating envelope. The wind-tunnel model was tested on a string and on a cable- mount system The model is shown on the cable-mount system with Langley engineer, Stanley Cole, checking tension in one of the support cables. All lifting surfaces were flutter cleared up to M=1.2 with the model string mounted. The model was then flutter cleared on the cable- mount system to assess the influence of rigid-body dynamics and fuselage flexibility on flutter. Several configuration parametric studies were also completed, including many external store configurations.

  17. Worst-Case Flutter Margins from F/A-18 Aircraft Aeroelastic Data

    NASA Technical Reports Server (NTRS)

    Lind, Rick; Brenner, Marty

    1997-01-01

    An approach for computing worst-case flutter margins has been formulated in a robust stability framework. Uncertainty operators are included with a linear model to describe modeling errors and flight variations. The structured singular value, micron, computes a stability margin which directly accounts for these uncertainties. This approach introduces a new method of computing flutter margins and an associated new parameter for describing these margins. The micron margins are robust margins which indicate worst-case stability estimates with respect to the defined uncertainty. Worst-case flutter margins are computed for the F/A-18 SRA using uncertainty sets generated by flight data analysis. The robust margins demonstrate flight conditions for flutter may lie closer to the flight envelope than previously estimated by p-k analysis.

  18. Control Law-Control Allocation Interaction: F/A-18 PA Simulation Test - Bed

    NASA Technical Reports Server (NTRS)

    Durham, Wayne; Nelson, Mark

    2001-01-01

    This report documents the first stage of research into Control Law - Control Allocation Interactions. A three-year research effort was originally proposed: 1. Create a desktop flight simulation environment under which experiments related to the open questions may be conducted. 2. Conduct research to determine which aspects of control allocation have impact upon control law design that merits further research. 3. Conduct research into those aspects of control allocation identified above, and their impacts upon control law design. Simulation code was written utilizing the F/A-18 airframe in the power approach (PA) configuration. A dynamic inversion control law was implemented and used to drive a state-of-the-art control allocation subroutine.

  19. Aeroelastic modeling for the FIT team F/A-18 simulation

    NASA Technical Reports Server (NTRS)

    Zeiler, Thomas A.; Wieseman, Carol D.

    1989-01-01

    Some details of the aeroelastic modeling of the F/A-18 aircraft done for the Functional Integration Technology (FIT) team's research in integrated dynamics modeling and how these are combined with the FIT team's integrated dynamics model are described. Also described are mean axis corrections to elastic modes, the addition of nonlinear inertial coupling terms into the equations of motion, and the calculation of internal loads time histories using the integrated dynamics model in a batch simulation program. A video tape made of a loads time history animation was included as a part of the oral presentation. Also discussed is work done in one of the areas of unsteady aerodynamic modeling identified as needing improvement, specifically, in correction factor methodologies for improving the accuracy of stability derivatives calculated with a doublet lattice code.

  20. Aeroelastic modeling for the FIT (Functional Integration Technology) team F/A-18 simulation

    NASA Technical Reports Server (NTRS)

    Zeiler, Thomas A.; Wieseman, Carol D.

    1989-01-01

    As part of Langley Research Center's commitment to developing multidisciplinary integration methods to improve aerospace systems, the Functional Integration Technology (FIT) team was established to perform dynamics integration research using an existing aircraft configuration, the F/A-18. An essential part of this effort has been the development of a comprehensive simulation modeling capability that includes structural, control, and propulsion dynamics as well as steady and unsteady aerodynamics. The structural and unsteady aerodynamics contributions come from an aeroelastic mode. Some details of the aeroelastic modeling done for the Functional Integration Technology (FIT) team research are presented. Particular attention is given to work done in the area of correction factors to unsteady aerodynamics data.

  1. Nakazawaea siamensis f.a., sp. nov., a yeast species isolated from phylloplane.

    PubMed

    Kaewwichian, Rungluk; Limtong, Savitree

    2014-01-01

    Strain DMKU-RK467(T), representing a novel yeast species, was isolated from the external surface of sugar cane leaves collected in Thailand. On the basis of morphological, biochemical, physiological and chemotaxonomic characteristics, and sequence analysis of the D1/D2 region of the LSU rRNA gene and the internal transcribed spacer (ITS) region, strain DMKU-RK467(T) was assigned to a novel species of the genus Nakazawaea. The novel species was related most closely to the type strain of Candida wickerhamii but they differed by 1.9 % nucleotide substitutions in the D1/D2 region of the LSU rRNA gene and by 5.2 % nucleotide substitutions in the ITS region. The name Nakazawaea siamensis f.a., sp. nov. is proposed (type strain DMKU-RK467(T) = BCC 50734(T) = NBRC 108903(T) = CBS 12569(T)). PMID:24052626

  2. Wing Torsional Stiffness Tests of the Active Aeroelastic Wing F/A-18 Airplane

    NASA Technical Reports Server (NTRS)

    Lokos, William A.; Olney, Candida D.; Crawford, Natalie D.; Stauf, Rick; Reichenbach, Eric Y.

    2002-01-01

    The left wing of the Active Aeroelastic Wing (AAW) F/A-18 airplane has been ground-load-tested to quantify its torsional stiffness. The test has been performed at the NASA Dryden Flight Research Center in November 1996, and again in April 2001 after a wing skin modification was performed. The primary objectives of these tests were to characterize the wing behavior before the first flight, and provide a before-and-after measurement of the torsional stiffness. Two streamwise load couples have been applied. The wing skin modification is shown to have more torsional flexibility than the original configuration has. Additionally, structural hysteresis is shown to be reduced by the skin modification. Data comparisons show good repeatability between the tests.

  3. Navier-Stokes Solutions about the F/A-18 Forebody-LEX Configuration

    NASA Technical Reports Server (NTRS)

    Ghaffari, Farhad; Bates, Brent L.; Luckring, James M.; Thomas, James L.

    1989-01-01

    Three-dimensional viscous flow computations are presented for the F/A-18 forebody-LEX geometry. Solutions are obtained from an algorithm for the compressible Navier-Stokes equations which incorporates an upwind-biased, flux-difference-splitting approach along with longitudinally-patched grids. Results are presented for both laminar and fully turbulent flow assumptions and include correlations with wind tunnel as well as flight-test results. A good quantitative agreement for the forebody surface pressure distribution is achieved between the turbulent computations and wind tunnel measurements at Mach number of 0.6. The computed turbulent surface flow patterns on the forebody qualitatively agree well with in-flight surface flow patterns obtained on an F/A-fS aircraft at Mach number of 0.34.

  4. Investigation of the Vortical Flow Above an F/A-18 Using Doppler Global Velocimetry

    NASA Technical Reports Server (NTRS)

    Meyers, James F.; Lee, Joseph W.; Cavone, Angelo A.; Suzuki, Karen E.

    1993-01-01

    The flow above an F/A-18 model, set to 25-degrees angle of attack, was measured using a Doppler global velocimeter, (DGV). The investigation indicated that the complex flow contained many similarities to the vortical flow above a simple delta wing set to a high-angle of attack, including flow standard deviations greater than 30-percent of free- stream. These standard deviation levels were also comparable to results found during a previous investigation of the vortical flow above a YF-17 using fringe-type laser velocimetry. The global measurement capability of the DGV provided the first evidence that the burst vortices above the model were structured. These structures were found to maintain their spatial coherence while the flow varied in an overall sense.

  5. Wind Tunnel Visualization of the Flow Over a Full-Scale F/A-18 Aircraft

    NASA Technical Reports Server (NTRS)

    Lanser, Wendy R.; Botha, Gavin J.; James, Kevin D.; Crowder, James P.; Schmitz, Fredric H. (Technical Monitor)

    1994-01-01

    The proposed paper presents flow visualization performed during experiments conducted on a full-scale F/A-18 aircraft in the 80- by 120-Foot Wind-Tunnel at NASA Ames Research Center. This investigation used both surface and off-surface flow visualization techniques to examine the flow field on the forebody, canopy, leading edge extensions (LEXs), and wings. The various techniques used to visualize the flow field were fluorescent tufts, flow cones treated with reflective material, smoke in combination with a laser light sheet, and a video imaging system. The flow visualization experiments were conducted over an angle of attack range from 20deg to 45deg and over a sideslip range from -10deg to 10deg. The results show regions of attached and separated flow on the forebody, canopy, and wings. Additionally, the vortical flow is clearly visible over the leading-edge extensions, canopy, and wings.

  6. Investigation of the vortical flow above an F/A-18 using Doppler global velocimetry

    NASA Astrophysics Data System (ADS)

    Myers, James F.; Lee, Joseph W.; Cavone, Angelo A.; Suzuki, Karen E.

    1993-08-01

    The flow above an F/A-18 model, set to 25-degrees angle of attack, was measured using a Doppler global velocimeter, (DGV). The investigation indicated that the complex flow contained many similarities to the vortical flow above a simple delta wing set to a high-angle of attack, including flow standard deviations greater than 30-percent of free-stream. These standard deviation levels were also comparable to results found during a previous investigation of the vortical flow above a YF-17 using fringe-type laser velocimetry. The global measurement capability of the DGV provided the first evidence that the burst vortices above the model were structured. These structures were found to maintain their spatial coherence while the flow varied in an overall sense.

  7. Comparing the performance of FA, DFA and DMA using different synthetic long-range correlated time series

    PubMed Central

    Shao, Ying-Hui; Gu, Gao-Feng; Jiang, Zhi-Qiang; Zhou, Wei-Xing; Sornette, Didier

    2012-01-01

    Notwithstanding the significant efforts to develop estimators of long-range correlations (LRC) and to compare their performance, no clear consensus exists on what is the best method and under which conditions. In addition, synthetic tests suggest that the performance of LRC estimators varies when using different generators of LRC time series. Here, we compare the performances of four estimators [Fluctuation Analysis (FA), Detrended Fluctuation Analysis (DFA), Backward Detrending Moving Average (BDMA), and Centred Detrending Moving Average (CDMA)]. We use three different generators [Fractional Gaussian Noises, and two ways of generating Fractional Brownian Motions]. We find that CDMA has the best performance and DFA is only slightly worse in some situations, while FA performs the worst. In addition, CDMA and DFA are less sensitive to the scaling range than FA. Hence, CDMA and DFA remain “The Methods of Choice” in determining the Hurst index of time series. PMID:23150785

  8. A general data-driven algorithm for façade structure modeling using ground based laser data

    NASA Astrophysics Data System (ADS)

    Yousefzadeh, M.; Leurink, F. H. M.; Beheshti jou, M.

    2014-08-01

    Façade reconstruction from laser point cloud has been an interesting subject in Photogrammetric community for the last two decades. However, due to the variety of architecture types and the nature of laser data, proposing a fully automatic modelling algorithm is still a challenge. Irregular architecture, density variation, occlusion and noise level are the main hindering factors of proposing a general model for façade reconstruction. This paper describes the sequences of an automatic data- driven method which starts from raw laser data and ends with object extraction. Statistical analysis was frequently utilized in segmentation, splitting line detection and object characterization. A rule-based modification method was employed to model the complexity of façade layout. Developed interface enables non-expert user to interact with modelling process by setting few parameters. The method was tested over a couple of datasets.

  9. Comparing the performance of FA, DFA and DMA using different synthetic long-range correlated time series.

    PubMed

    Shao, Ying-Hui; Gu, Gao-Feng; Jiang, Zhi-Qiang; Zhou, Wei-Xing; Sornette, Didier

    2012-01-01

    Notwithstanding the significant efforts to develop estimators of long-range correlations (LRC) and to compare their performance, no clear consensus exists on what is the best method and under which conditions. In addition, synthetic tests suggest that the performance of LRC estimators varies when using different generators of LRC time series. Here, we compare the performances of four estimators [Fluctuation Analysis (FA), Detrended Fluctuation Analysis (DFA), Backward Detrending Moving Average (BDMA), and Centred Detrending Moving Average (CDMA)]. We use three different generators [Fractional Gaussian Noises, and two ways of generating Fractional Brownian Motions]. We find that CDMA has the best performance and DFA is only slightly worse in some situations, while FA performs the worst. In addition, CDMA and DFA are less sensitive to the scaling range than FA. Hence, CDMA and DFA remain "The Methods of Choice" in determining the Hurst index of time series. PMID:23150785

  10. The polygalacturonase FaPG1 gene plays a key role in strawberry fruit softening.

    PubMed

    García-Gago, Juan A; Posé, Sara; Muñoz-Blanco, Juan; Quesada, Miguel A; Mercado, José A

    2009-08-01

    The loss of firm texture is one of the most characteristic physiological processes that occur during the ripening of fleshy fruits. It is generally accepted that the disassembly of primary cell wall and middle lamella is the main factor involved in fruit softening. In this process, polygalacturonase (PG) has been implicated in the degradation of the polyuronide network in several fruits. However, the minor effect of PG downregulation on tomato softening, reported during the nineties, minimized the role of this enzyme in softening. Further works in other fruits are challenging this general assumption, as is occurring in strawberry. The strawberry (Fragaria x ananassa) fruit undergoes an extensive and fast softening that limit its shelf life and postharvest. Traditionally, it has also been considered that PG plays a minor role on this process, due to the low PG activity found in ripened strawberry fruits. Transgenic strawberry plants expressing an antisense sequence of the ripening-specific PG gene FaPG1 have been generated to get an insight into the role of this gene in softening. Half of the transgenic lines analyzed yielded fruits significantly firmer than control, without being affected other fruit parameters such as weight, color or soluble solids. The increase on firmness was maintained after several days of posharvest. In these firmer lines, FaPG1 was silenced to 95%, but total PG activity was only minor reduced. At the cell wall level, transgenic fruits contained a higher amount of covalently bound pectins whereas the soluble fraction was diminished. A microarray analysis of genes expressed in ripened receptacle did not show any significant change between control and transgenic fruits. Thus, contrary to the most accepted view, it is concluded that PG plays a key role on pectin metabolism and softening of strawberry fruit. PMID:19820312

  11. Rapid evaluation and quality control of next generation sequencing data with FaQCs

    DOE PAGESBeta

    Lo, Chien -Chi; Chain, Patrick S. G.

    2014-12-01

    Background: Next generation sequencing (NGS) technologies that parallelize the sequencing process and produce thousands to millions, or even hundreds of millions of sequences in a single sequencing run, have revolutionized genomic and genetic research. Because of the vagaries of any platform's sequencing chemistry, the experimental processing, machine failure, and so on, the quality of sequencing reads is never perfect, and often declines as the read is extended. These errors invariably affect downstream analysis/application and should therefore be identified early on to mitigate any unforeseen effects. Results: Here we present a novel FastQ Quality Control Software (FaQCs) that can rapidly processmore » large volumes of data, and which improves upon previous solutions to monitor the quality and remove poor quality data from sequencing runs. Both the speed of processing and the memory footprint of storing all required information have been optimized via algorithmic and parallel processing solutions. The trimmed output compared side-by-side with the original data is part of the automated PDF output. We show how this tool can help data analysis by providing a few examples, including an increased percentage of reads recruited to references, improved single nucleotide polymorphism identification as well as de novo sequence assembly metrics. Conclusion: FaQCs combines several features of currently available applications into a single, user-friendly process, and includes additional unique capabilities such as filtering the PhiX control sequences, conversion of FASTQ formats, and multi-threading. The original data and trimmed summaries are reported within a variety of graphics and reports, providing a simple way to do data quality control and assurance.« less

  12. Rapid evaluation and quality control of next generation sequencing data with FaQCs

    SciTech Connect

    Lo, Chien -Chi; Chain, Patrick S. G.

    2014-12-01

    Background: Next generation sequencing (NGS) technologies that parallelize the sequencing process and produce thousands to millions, or even hundreds of millions of sequences in a single sequencing run, have revolutionized genomic and genetic research. Because of the vagaries of any platform's sequencing chemistry, the experimental processing, machine failure, and so on, the quality of sequencing reads is never perfect, and often declines as the read is extended. These errors invariably affect downstream analysis/application and should therefore be identified early on to mitigate any unforeseen effects. Results: Here we present a novel FastQ Quality Control Software (FaQCs) that can rapidly process large volumes of data, and which improves upon previous solutions to monitor the quality and remove poor quality data from sequencing runs. Both the speed of processing and the memory footprint of storing all required information have been optimized via algorithmic and parallel processing solutions. The trimmed output compared side-by-side with the original data is part of the automated PDF output. We show how this tool can help data analysis by providing a few examples, including an increased percentage of reads recruited to references, improved single nucleotide polymorphism identification as well as de novo sequence assembly metrics. Conclusion: FaQCs combines several features of currently available applications into a single, user-friendly process, and includes additional unique capabilities such as filtering the PhiX control sequences, conversion of FASTQ formats, and multi-threading. The original data and trimmed summaries are reported within a variety of graphics and reports, providing a simple way to do data quality control and assurance.

  13. [Risk of nasopharyngeal cancer, Leukemia and other tumors in a cohort of employees and students potentially exposed to (FA) formaldehyde in University laboratories].

    PubMed

    Sernia, S; Di Folco, F; Altrudo, P; Sbriccoli, B; Sestili, C; Colamesta, V; Del Buono, S; Michetti, A; Ortis, M; Dawodu, A; Villari, P; La Torre, G

    2016-01-01

    FA was recently classified as carcinogen of second class (category 1B). A retrospective cohort study was conducted for the evaluation of the association between exposure to FA and cancer in professionally potentially exposed in a University setting. The cohort was composed of 140 exposed to FA and 364 not exposed in the period 1999-2015. The results showed no cancers of naso-pharynx and leukemias or lymphomas both among exposed and not exposed. Moreover, the exposure to FA is not significantly associated to an increase of other types of tumors. PMID:27212573

  14. The Oncogenic Small Tumor Antigen of Merkel Cell Polyomavirus Is an Iron-Sulfur Cluster Protein That Enhances Viral DNA Replication

    PubMed Central

    Tsang, Sabrina H.; Wang, Ranran; Nakamaru-Ogiso, Eiko; Knight, Simon A. B.; Buck, Christopher B.

    2015-01-01

    ABSTRACT Merkel cell polyomavirus (MCPyV) plays an important role in Merkel cell carcinoma (MCC). MCPyV small T (sT) antigen has emerged as the key oncogenic driver in MCC carcinogenesis. It has also been shown to promote MCPyV LT-mediated replication by stabilizing LT. The importance of MCPyV sT led us to investigate sT functions and to identify potential ways to target this protein. We discovered that MCPyV sT purified from bacteria contains iron-sulfur (Fe/S) clusters. Electron paramagnetic resonance analysis showed that MCPyV sT coordinates a [2Fe-2S] and a [4Fe-4S] cluster. We also observed phenotypic conservation of Fe/S coordination in the sTs of other polyomaviruses. Since Fe/S clusters are critical cofactors in many nucleic acid processing enzymes involved in DNA unwinding and polymerization, our results suggested the hypothesis that MCPyV sT might be directly involved in viral replication. Indeed, we demonstrated that MCPyV sT enhances LT-mediated replication in a manner that is independent of its previously reported ability to stabilize LT. MCPyV sT translocates to nuclear foci containing actively replicating viral DNA, supporting a direct role for sT in promoting viral replication. Mutations of Fe/S cluster-coordinating cysteines in MCPyV sT abolish its ability to stimulate viral replication. Moreover, treatment with cidofovir, a potent antiviral agent, robustly inhibits the sT-mediated enhancement of MCPyV replication but has little effect on the basal viral replication driven by LT alone. This finding further indicates that MCPyV sT plays a direct role in stimulating viral DNA replication and introduces cidofovir as a possible drug for controlling MCPyV infection. IMPORTANCE MCPyV is associated with a highly aggressive form of skin cancer in humans. Epidemiological surveys for MCPyV seropositivity and sequencing analyses of healthy human skin suggest that MCPyV may represent a common component of the human skin microbial flora. However, much of the

  15. Genetic Analysis of Strawberry Fruit Aroma and Identification of O-Methyltransferase FaOMT as the Locus Controlling Natural Variation in Mesifurane Content1[C][W][OA

    PubMed Central

    Zorrilla-Fontanesi, Yasmín; Rambla, José-Luis; Cabeza, Amalia; Medina, Juan J.; Sánchez-Sevilla, José F.; Valpuesta, Victoriano; Botella, Miguel A.; Granell, Antonio; Amaya, Iraida

    2012-01-01

    Improvement of strawberry (Fragaria × ananassa) fruit flavor is an important goal in breeding programs. To investigate genetic factors controlling this complex trait, a strawberry mapping population derived from genotype ‘1392’, selected for its superior flavor, and ‘232’ was profiled for volatile compounds over 4 years by headspace solid phase microextraction coupled to gas chromatography and mass spectrometry. More than 300 volatile compounds were detected, of which 87 were identified by comparison of mass spectrum and retention time to those of pure standards. Parental line ‘1392’ displayed higher volatile levels than ‘232’, and these and many other compounds with similar levels in both parents segregated in the progeny. Cluster analysis grouped the volatiles into distinct chemically related families and revealed a complex metabolic network underlying volatile production in strawberry fruit. Quantitative trait loci (QTL) detection was carried out over 3 years based on a double pseudo-testcross strategy. Seventy QTLs covering 48 different volatiles were detected, with several of them being stable over time and mapped as major QTLs. Loci controlling γ-decalactone and mesifurane content were mapped as qualitative traits. Using a candidate gene approach we have assigned genes that are likely responsible for several of the QTLs. As a proof of concept we show that one homoeolog of the O-methyltransferase gene (FaOMT) is the locus responsible for the natural variation of mesifurane content. Sequence analysis identified 30 bp in the promoter of this FaOMT homoeolog containing putative binding sites for basic/helix-loop-helix, MYB, and BZIP transcription factors. This polymorphism fully cosegregates with both the presence of mesifurane and the high expression of FaOMT during ripening. PMID:22474217

  16. CARTILAGE CELL CLUSTERS

    PubMed Central

    Lotz, Martin K.; Otsuki, Shuhei; Grogan, Shawn P.; Sah, Robert; Terkeltaub, Robert; D’Lima, Darryl

    2010-01-01

    The formation of new cell clusters is a histological hallmark of arthritic cartilage but the biology of clusters and their role in disease are poorly understood. This is the first comprehensive review of clinical and experimental conditions associated with cluster formation. Genes and proteins that are expressed in cluster cells, the cellular origin of the clusters, mechanisms that lead to cluster formation and the role of cluster cells in pathogenesis are discussed. PMID:20506158

  17. Viewing time measures of sexual orientation in Samoan cisgender men who engage in sexual interactions with fa'afafine.

    PubMed

    Petterson, Lanna J; Dixson, Barnaby J; Little, Anthony C; Vasey, Paul L

    2015-01-01

    Androphilia refers to attraction to adult males, whereas gynephilia refers to attraction to adult females. The current study employed self-report and viewing time (response time latency) measures of sexual attraction to determine the sexual orientation of Samoan cisgender men (i.e., males whose gender presentation and identity is concordant with their biological sex) who engage in sexual interactions with transgender male androphiles (known locally as fa'afafine) compared to: (1) Samoan cisgender men who only engage in sexual interactions with women, and (2) fa'afafine. As expected, both measures indicated that cisgender men who only engaged in sexual interactions with women exhibited a gynephilic pattern of sexual attraction, whereas fa'afafine exhibited an androphilic one. In contrast, both measures indicated that cisgender men who engaged in sexual interactions with fa'afafine demonstrated a bisexual pattern of sexual attraction. Most of the cisgender men who exhibited bisexual viewing times did not engage in sexual activity with both men and women indicating that the manner in which bisexual patterns of sexual attraction manifest behaviorally vary from one culture to the next. PMID:25679961

  18. Behind the Façade of Fee-Free education: Shadow Education and Its Implications for Social Justice

    ERIC Educational Resources Information Center

    Bray, Mark; Kwo, Ora

    2013-01-01

    Most governments, at an official level, espouse the principles of the 1948 Universal Declaration of Human Rights. Among its statements is that education shall be free, at least in the elementary and fundamental stages. Yet while the façade of government education systems presents an image that instruction is free of charge, families across the…

  19. Production Support Flight Control Computers: Research Capability for F/A-18 Aircraft at Dryden Flight Research Center

    NASA Technical Reports Server (NTRS)

    Carter, John F.

    1997-01-01

    NASA Dryden Flight Research Center (DFRC) is working with the United States Navy to complete ground testing and initiate flight testing of a modified set of F/A-18 flight control computers. The Production Support Flight Control Computers (PSFCC) can give any fleet F/A-18 airplane an in-flight, pilot-selectable research control law capability. NASA DFRC can efficiently flight test the PSFCC for the following four reasons: (1) Six F/A-18 chase aircraft are available which could be used with the PSFCC; (2) An F/A-18 processor-in-the-loop simulation exists for validation testing; (3) The expertise has been developed in programming the research processor in the PSFCC; and (4) A well-defined process has been established for clearing flight control research projects for flight. This report presents a functional description of the PSFCC. Descriptions of the NASA DFRC facilities, PSFCC verification and validation process, and planned PSFCC projects are also provided.

  20. 2 CFR 200.411 - Adjustment of previously negotiated indirect (F&A) cost rates containing unallowable costs.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... by the cognizant agency for indirect costs. The choice of method must be at the discretion of the cognizant agency for indirect costs, based on its judgment as to which method would be most practical. (e... Cost Principles Basic Considerations § 200.411 Adjustment of previously negotiated indirect (F&A)...

  1. Determination of the refractive index difference caused by the birefringence of FA (II) centers in KCl:Li

    NASA Astrophysics Data System (ADS)

    Silfsten, Pertti; Ketolainen, Pertti

    1991-11-01

    A method is described for determining the refractive index difference caused by the birefringence of oriented FA (II) centers in KCl:Li crystals. It is shown that the portion induced by the birefringence can be separated from an absorption spectrum measured through a polarizer-analyzer system. From this portion the refractive index difference can then be calculated with ease.

  2. The optimization of FA/O barrier slurry with respect to removal rate selectivity on patterned Cu wafers

    NASA Astrophysics Data System (ADS)

    Yi, Hu; Yan, Li; Yuling, Liu; Yangang, He

    2016-02-01

    Because the polishing of different materials is required in barrier chemical mechanical planarization (CMP) processes, the development of a kind of barrier slurry with improved removal rate selectivity for Cu/barrier/TEOS would reduce erosion and dishing defects on patterned Cu wafers. In this study, we developed a new benzotriazole-free barrier slurry named FA/O barrier slurry, containing 20 mL/L of the chelating agent FA/O, 5 mL/L surfactant, and a 1:5 concentration of abrasive particles. By controlling the polishing slurry ingredients, the removal rate of different materials could be controlled. For process integration considerations, the effect of the FA/O barrier slurry on the dielectric layer of the patterned Cu wafer was investigated. After CMP processing by the FA/O barrier slurry, the characteristics of the dielectric material were tested. The results showed that the dielectric characteristics met demands for industrial production. The current leakage was of pA scale. The resistance and capacitance were 2.4 kω and 2.3 pF, respectively. The dishing and erosion defects were both below 30 nm in size. CMP-processed wafers using this barrier slurry could meet industrial production demands. Project supported by the Special Project Items No. 2 in National Long-Term Technology Development Plan (No. 2009ZX02308), the Natural Science Foundation of Hebei Province (No. F2012202094), and the Doctoral Program Foundation of Xinjiang Normal University Plan (No. XJNUBS1226).

  3. A NASA F/A-18, participating in the Automated Aerial Refueling (AAR) project, flies over the Dryden

    NASA Technical Reports Server (NTRS)

    2002-01-01

    A NASA F/A-18 is participating in the Automated Aerial Refueling (AAR) project. The 300-gallon aerial refueling store seen on the belly of the aircraft carries fuel and a refueling drogue. This aircraft acts as a tanker in the study to develop an aerodynamic model for future automated aerial refueling, especially of unmanned vehicles.

  4. A Hypomorphic PALB2 Allele Gives Rise to an Unusual Form of FA-N Associated with Lymphoid Tumour Development

    PubMed Central

    Byrd, Philip J.; Stewart, Grant. S.; Smith, Anna; Eaton, Charlotte; Taylor, Alexander J.; Guy, Chloe; Eringyte, Ieva; Fooks, Peggy; Last, James I.; Horsley, Robert; Oliver, Antony W.; Janic, Dragana; Dokmanovic, Lidija; Stankovic, Tatjana; Taylor, A. Malcolm R.

    2016-01-01

    Patients with biallelic truncating mutations in PALB2 have a severe form of Fanconi anaemia (FA-N), with a predisposition for developing embryonal-type tumours in infancy. Here we describe two unusual patients from a single family, carrying biallelic PALB2 mutations, one truncating, c.1676_1677delAAinsG;(p.Gln559ArgfsTer2), and the second, c.2586+1G>A; p.Thr839_Lys862del resulting in an in frame skip of exon 6 (24 amino acids). Strikingly, the affected individuals did not exhibit the severe developmental defects typical of FA-N patients and initially presented with B cell non-Hodgkin lymphoma. The expressed p.Thr839_Lys862del mutant PALB2 protein retained the ability to interact with BRCA2, previously unreported in FA-N patients. There was also a large increased chromosomal radiosensitivity following irradiation in G2 and increased sensitivity to mitomycin C. Although patient cells were unable to form Rad51 foci following exposure to either DNA damaging agent, U2OS cells, in which the mutant PALB2 with in frame skip of exon 6 was induced, did show recruitment of Rad51 to foci following damage. We conclude that a very mild form of FA-N exists arising from a hypomorphic PALB2 allele. PMID:26990772

  5. Inductions of the fatty acid 2-hydroxylase (FA2H) gene by Δ9-tetrahydrocannabinol in human breast cancer cells

    PubMed Central

    Takeda, Shuso; Harada, Mari; Su, Shengzhong; Okajima, Shunsuke; Miyoshi, Hiroko; Yoshida, Kazutaka; Nishimura, Hajime; Okamoto, Yoshiko; Amamoto, Toshiaki; Watanabe, Kazuhito; Omiecinski, Curtis J; Aramaki, Hironori

    2014-01-01

    To investigate gene(s) being regulated by Δ9-tetrahydrocannabinol (Δ9-THC), we performed DNA microarray analysis of human breast cancer MDA-MB-231 cells, which are poorly differentiated breast cancer cells, treated with Δ9-THC for 48 hr at an IC50 concentration of approximately 25 μM. Among the highly up-regulated genes (> 10-fold) observed, fatty acid 2-hydroxylase (FA2H) was significantly induced (17.8-fold). Although the physiological role of FA2H has not yet been fully understood, FA2H has been shown to modulate cell differentiation. The results of Oil Red O staining after Δ9-THC exposure showed the distribution of lipid droplets (a sign of the differentiated phenotype) in cells. Taken together, the results obtained here indicate that FA2H is a novel Δ9-THC-regulated gene, and that Δ9-THC induces differentiation signal(s) in poorly differentiated MDA-MB-231 cells. PMID:23535410

  6. Water-tunnel study results of a TF/A-18 and F/A-18 canopy flow visualization

    NASA Technical Reports Server (NTRS)

    Johnson, Steven A.; Fisher, David F.

    1990-01-01

    A water tunnel study examining the influence of canopy shape on canopy and leading edge extension flow patterns was initiated. The F/A-18 single-place canopy model and the TF/A-18 two place canopy model were the study subjects. Plan view and side view photographs showing the flow patterns created by injected colored dye are presented for 0 deg and 5 deg sideslip angles. Photographs taken at angle of attack and sideslip conditions correspond to test departure points found in flight test. Flight experience has shown that the TF/A-18 airplane departs in regions where the F/A-18 airplane is departure-resistant. The study results provide insight into the differences in flow patterns which may influence the resulting aerodynamics of the TF/A-18 and F/A-18 aircraft. It was found that at 0 deg sideslip, the TF/A-18 model has more downward flow on the sides of the canopy than the F/A-18 model. This could be indicative of flow from the leading edge extension (LEX) vortexes impinging on the sides of the wider TF/A-18 canopy. In addition, the TF/A-18 model has larger areas of asymmetric separated and unsteady flow on the LEXs and fuselage, possibly indicating a lateral and directional destabilizing effect at the conditions studied.

  7. Evaluation of F/A-18A HARV inlet flow analysis with flight data

    NASA Technical Reports Server (NTRS)

    Smith, C. Frederic; Podleski, Steve D.; Barankiewicz, Wendy S.; Zeleznik, Susan Z.

    1995-01-01

    The F/A-18A aircraft has experienced engine stalls at high angles-of-attack and yaw flight conditions which were outside of its flight envelope. Future aircraft may be designed to operate routinely in this flight regime. Therefore, it is essential that an understanding of the inlet flow field at these flight conditions be obtained. Due to the complex interactions of the fuselage and inlet flow fields, a study of the flow within the inlet must also include external effects. Full Navier-Stokes (FNS) calculations on the F/A-18A High Alpha Research Vehicle (HARV) inlet for several angles-of-attack with sideslip and free stream Mach numbers have been obtained. The predicted forebody/fuselage surface static pressures agreed well with flight data. The surface static pressures along the inlet lip are in good agreement with the numerical predictions. The major departure in agreement is along the bottom of the lip at 30 deg and 60 deg angle-of-attack where a possible streamwise flow separation is not being predicted by the code. The circumferential pressure distributions at the engine face are in very good agreement with the numerical results. The variation in surface static pressure in the circumferential direction is very small with the exception of 60 angle-of-attack. Although the simulation does not include the effect of the engine, it appears that this omission has a second order effect on the circumferential pressure distribution. An examination of the unsteady flight test data base has shown that the secondary vortex migrates a significant distance with time. In fact, the extent of this migration increases with angle-of-attack with increasing levels of distortion. The effects of the engine on this vortex movement is unknown. This implies that the level of flow unsteadiness increases with increasing distortion. Since the computational results represent an asymptotic solution driven by steady boundary conditions, these numerical results may represent an arbitrary point

  8. Environmental and health effects of nanomaterials in nanotextiles and façade coatings.

    PubMed

    Som, Claudia; Wick, Peter; Krug, Harald; Nowack, Bernd

    2011-08-01

    Engineered nanomaterials (ENM) are expected to hold considerable potential for products that offer improved or novel functionalities. For example, nanotechnologies could open the way for the use of textile products outside their traditional fields of applications, for example, in the construction, medical, automobile, environmental and safety technology sectors. Consequently, nanotextiles could become ubiquitous in industrial and consumer products in future. Another ubiquitous field of application for ENM is façade coatings. The environment and human health could be affected by unintended release of ENM from these products. The product life cycle and the product design determine the various environmental and health exposure situations. For example, ENM unintentionally released from geotextiles will probably end up in soils, whereas ENM unintentionally released from T-shirts may come into direct contact with humans and end up in wastewater. In this paper we have assessed the state of the art of ENM effects on the environment and human health on the basis of selected environmental and nanotoxicological studies and on our own environmental exposure modeling studies. Here, we focused on ENM that are already applied or may be applied in future to textile products and façade coatings. These ENM's are mainly nanosilver (nano-Ag), nano titanium dioxide (nano-TiO(2)), nano silica (nano-SiO(2)), nano zinc oxide (nano-ZnO), nano alumina (nano-Al(2)O(3)), layered silica (e.g. montmorillonite, Al(2)[(OH)(2)/Si(4)O(10)]nH(2)O), carbon black, and carbon nanotubes (CNT). Knowing full well that innovators have to take decisions today, we have presented some criteria that should be useful in systematically analyzing and interpreting the state of the art on the effects of ENM. For the environment we established the following criteria: (1) the indication for hazardous effects, (2) dissolution in water increases/decreases toxic effects, (3) tendency for agglomeration or sedimentation

  9. Lovastatin causes FaDu hypopharyngeal carcinoma cell death via AMPK-p63-survivin signaling cascade

    PubMed Central

    Yen, Chia-Sheng; Chen, Jung-Chien; Chang, Yi-Fang; Hsu, Ya-Fen; Chiu, Pei-Ting; Shiue, Ching; Chuang, Yu-Fan; Ou, George; Hsu, Ming-Jen

    2016-01-01

    Statins are used widely to lower serum cholesterol and the incidence of cardiovascular diseases. Growing evidence shows that statins also exhibit beneficial effects against cancers. In this study, we investigated the molecular mechanisms involved in lovastatin-induced cell death in Fadu hypopharyngeal carcinoma cells. Lovastatin caused cell cycle arrest and apoptosis in FaDu cells. Lovastatin increased p21cip/Waf1 level while the survivin level was decreased in the presence of lovastatin. Survivin siRNA reduced cell viability and induced cell apoptosis in FaDu cells. Lovastatin induced phosphorylation of AMP-activated protein kinase (AMPK), p38 mitogen-activated protein kinase (MAPK) and transcription factor p63. Lovastatin also caused p63 acetylation and increased p63 binding to survivin promoter region in FaDu cells. AMPK-p38MAPK signaling blockade abrogated lovastatin-induced p63 phosphorylation. Lovastatin’s enhancing effect on p63 acetylation was reduced in HDAC3- or HDAC4- transfected cells. Moreover, transfection of cells with AMPK dominant negative mutant (AMPK-DN), HDAC3, HDAC4 or p63 siRNA significantly reduced lovastatin’s effects on p21cip/Waf1 and survivin. Furthermore, lovastatin inhibited subcutaneous FaDu xenografts growth in vivo. Taken together, lovastatin may activate AMPK-p38MAPK-p63-survivin cascade to cause FaDu cell death. This study establishes, at least in part, the signaling cascade by which lovastatin induces hypopharyngeal carcinoma cell death. PMID:27122225

  10. Effect of celecoxib on inhibiting tumor repopulation during radiotherapy in human FaDu squamous cell carcinoma

    PubMed Central

    Yang, Jia; Liu, Jing; Sun, Xin-Dong; Hu, Xu-Dong; Sun, Ju-Jie; Li, Yu-Hui; Yu, Jin-Ming

    2014-01-01

    Aim of the study FaDu human squamous cell carcinoma (FaDu-hSCC) demonstrated accelerated tumor repopulation during fractionated irradiation with pathological validation in a xenograft model system. Previous studies showed that the selective cyclooxygenase (COX)-2 inhibitor celecoxib can enhance the tumor response to radiotherapy. So we aimed to explore the effect of celecoxib in inducing apoptosis and inhibiting repopulation of FaDu tumors in nude mice during fractionated radiotherapy. Material and methods FaDu-hSCC was transplanted into the right hind leg of BALB/C nude mice. Mice were treated with celecoxib and/or fractionated irradiation. Celecoxib (100 mg/kg/day) was administered by daily gavage. Irradiation was delivered with 12 to 18 fractions of 3.0 Gy daily or every second day based on Petersen's repopulation model. At different time points, tumors were excised for immunohistochemistry staining. Results Significant tumor repopulation occurred after about 18 days of radiotherapy. On average, Ki-67 and bromodeoxyuridine (BrdUrd) labeling indices (LI) decreased with daily irradiation (both p < 0.05) and increased with every-second-day irradiation (both p > 0.05), suggesting accelerated repopulation. Ki-67 LI decreased in celecoxib concurrent with radiotherapy for 12 fractions in 24 days and 18 fractions in 36 days compared with irradiated alone (p = 0.004 and 0.042, respectively). BrdUrd LI values were lower in the concurrent groups than irradiated alone (p = 0.001 and 0.006, respectively). Epithelial growth factor receptor (EGFR) expression score decreased in the concurrent groups than irradiated alone (p = 0.037 and 0.031, respectively). Caspase-3 expression scores were higher in the concurrent groups than irradiated alone (p = 0.05 and 0.006, respectively). Conclusions Celecoxib concurrent radiotherapy could inhibit tumor repopulation and increase tumor apoptosis during the treatment in FaDu squamous cell carcinoma. PMID:25258584

  11. Update: HFC 245fa Blown Foam Development with External Tank Spray Foams

    NASA Technical Reports Server (NTRS)

    Davis, S.

    2001-01-01

    The Marshall Space Flight Center (MSFC) Thermal Protection Systems (TPS) Materials Research Laboratory is currently investigating environmentally friendly blowing agents for use in the insulations of the Space Shuttle's External Tank. The original TPS foam materials of the External Tank were blown with chlorofluorocarbon 11, which is now regulated because of its high Ozone Depletion Potential (ODP). Hydrochlorofluorocarbons (HCFCs), with an ODP that is one tenth that of CFCs, have been widely adopted as an interim blowing agent in urethane insulations. In FY96, Lockheed Martin completed the production qualification and validation of HCFC 141b blown insulations. Because of the expected limited commercial lifetime of HCFC 141b, research efforts are underway to identify and develop alternatives with zero ODP. HFC245fa (1,1,1,3,3-pentaflouropropane) has been chosen by the manufacturer as a third-generation blowing agent to be marketed commercially. Preliminary work evaluating this third-generation candidate has demonstrated promising material mechanical property data. Favorable results from small-scale spray activities have justified evaluations using production foam processing spray parameters. With the scale-up of the spray equipment, however, additional processing issues have been identified. This paper will present data collected to date regarding the use of this blowing agent in External Tank spray foams.

  12. F/A-18 forebody vortex control. Volume 2: Rotary-balance tests

    NASA Technical Reports Server (NTRS)

    Kramer, Brian R.; Suarez, Carlos J.; Malcolm, Gerald N.; Ayers, Bert F.

    1994-01-01

    A rotary-balance wind tunnel test was conducted on a six percent model of the F/A-18 at the NASA Ames 7 X 10-Foot Low Speed Wind Tunnel. The data reduction was specially written for the test in National Instruments' LabVIEW. The data acquisition, reduction and analysis was performed with a Macintosh computer. The primary objective of the test was to evaluate the effectiveness of several forebody vortex control configurations in a rotary flow field. The devices that were found to be the most effective during the static tests (Volume 1) were investigated and included both mechanical and pneumatic configurations. The mechanical systems evaluated were small, single and dual, rotating nose tip strakes and a vertical nose strake. The jet blowing configuration used nozzles canted inboard 60 degrees. A two segment tangential slot was also evaluated. The different techniques were evaluated at angles of attack of 30 degrees, 45 degrees, 51 degrees, and 60 degrees. Sideslip and Reynolds number were varied for some of the configurations. All of the techniques proved to be effective in the rotating flow field. The vertical nose strake had the largest 'envelope' of effectiveness. Forebody vortex control provides large, robust yawing moments at medium to high angles of attack, even during combat maneuvers such as loaded roll.

  13. Full-scale high angle-of-attack tests of an F/A-18

    NASA Technical Reports Server (NTRS)

    Meyn, Larry A.; Lanser, Wendy R.; James, Kevin D.

    1992-01-01

    This paper presents an overview of high angle-of-attack tests of a full-scale F/A-18 in the 80- by 120-Foot Wind Tunnel of the National Full-Scale Aerodynamic Complex at NASA Ames Research Center at Moffett Field, California. A production aircraft was tested over an angle-of-attack range of 18 to 50 deg and at wind speeds of up to 100 knots. These tests had three primary test objectives. Pneumatic and mechanical forebody flow control devices were tested at full-scale and shown to produce significant yawing moments for lateral control of the aircraft at high angles of attack. Mass flow requirements for the pneumatic system were found to scale with freestream density and speed rather than freestream dynamic pressure. Detailed measurements of the pressures buffeting the vertical tail were made and spatial variations in the buffeting frequency were found. The LEX fence was found to have a significant effect on the frequency distribution on the outboard surface of the vertical fin. In addition to the above measurements, an extensive set of data was acquired for the validation of computational fluid dynamics codes and for comparison with flight test and small-scale wind tunnel test results.

  14. Active Control of F/A-18 Vertical Tail Buffeting using Piezoelectric Actuators

    NASA Technical Reports Server (NTRS)

    Sheta, Essam F.; Moses, Robert W.; Huttsell, Lawerence J.; Harrand, Vincent J.

    2003-01-01

    Vertical tail buffeting is a serious multidisciplinary problem that limits the performance of twin-tail fighter aircraft. The buffet problem occurs at high angles of attack when the vortical flow breaks down ahead of the vertical tails resulting in unsteady and unbalanced pressure loads on the vertical tails. This paper describes a multidisciplinary computational investigation for buffet load alleviation of full F/A-18 aircraft using distributed piezoelectric actuators. The inboard and outboard surfaces of the vertical tail are equipped with piezoelectric actuators to control the buffet responses in the first bending and torsion modes. The electrodynamics of the smart structure are expressed with a three-dimensional finite element model. A single-input-single-output controller is designed to drive the active piezoelectric actuators. High-fidelity multidisciplinary analysis modules for the fluid dynamics, structure dynamics, electrodynamics of the piezoelectric actuators, fluid-structure interfacing, and grid motion are integrated into a multidisciplinary computing environment that controls the temporal synchronization of the analysis modules. Peak values of the power spectral density of tail tip acceleration are reduced by as much as 22% in the first bending mode and by as much as 82% in the first torsion mode. RMS values of tip acceleration are reduced by as much as 12%.

  15. F/A-18 1/9th scale model tail buffet measurements

    NASA Technical Reports Server (NTRS)

    Martin, C. A.; Glaister, M. K.; Maclaren, L. D.; Meyn, L. A.; Ross, J.

    1991-01-01

    Wind tunnel tests were carried out on a 1/9th scale model of the F/A-18 at high angles of attack to investigate the characteristics of tail buffet due to bursting of the wing leading edge extension (LEX) vortices. The tests were carried out at the Aeronautical Research Laboratory low-speed wind tunnel facility and form part of a collaborative activity with NASA Ames Research Center, organized by The Technical Cooperative Program (TTCP). Information from the program will be used in the planning of similar collaborative tests, to be carried out at NASA Ames, on a full-scale aircraft. The program covered the measurement of unsteady pressures and fin vibration for cases with and without the wing LEX fences fitted. Fourier transform methods were used to analyze the unsteady data, and information on the spatial and temporal content of the vortex burst pressure field was obtained. Flow visualization of the vortex behavior was carried out using smoke and a laser light sheet technique.

  16. Adaptive Augmenting Control Flight Characterization Experiment on an F/A-18

    NASA Technical Reports Server (NTRS)

    VanZwieten, Tannen S.; Gilligan, Eric T.; Wall, John H.; Orr, Jeb S.; Miller, Christopher J.; Hanson, Curtis E.

    2014-01-01

    The NASA Marshall Space Flight Center (MSFC) Flight Mechanics and Analysis Division developed an Adaptive Augmenting Control (AAC) algorithm for launch vehicles that improves robustness and performance by adapting an otherwise welltuned classical control algorithm to unexpected environments or variations in vehicle dynamics. This AAC algorithm is currently part of the baseline design for the SLS Flight Control System (FCS), but prior to this series of research flights it was the only component of the autopilot design that had not been flight tested. The Space Launch System (SLS) flight software prototype, including the adaptive component, was recently tested on a piloted aircraft at Dryden Flight Research Center (DFRC) which has the capability to achieve a high level of dynamic similarity to a launch vehicle. Scenarios for the flight test campaign were designed specifically to evaluate the AAC algorithm to ensure that it is able to achieve the expected performance improvements with no adverse impacts in nominal or nearnominal scenarios. Having completed the recent series of flight characterization experiments on DFRC's F/A-18, the AAC algorithm's capability, robustness, and reproducibility, have been successfully demonstrated. Thus, the entire SLS control architecture has been successfully flight tested in a relevant environment. This has increased NASA's confidence that the autopilot design is ready to fly on the SLS Block I vehicle and will exceed the performance of previous architectures.

  17. Inlet Flow Characteristics During Rapid Maneuvers for an F/A-18A Airplane

    NASA Technical Reports Server (NTRS)

    Steenken, William G.; Williams, John G.; Walsh, Kevin R.

    1999-01-01

    The F404-GE-400 engine powered F/A-18A High Alpha Research Vehicle (HARV) was used to examine the characteristics of inlet airflow during rapid aircraft maneuvers. A study of the degree of similarity between inlet data obtained during rapid aircraft maneuvers and inlet data obtained at steady aerodynamic attitudes was conducted at the maximum engine airflow of approximately 145 Ibm/sec using a computer model that was generated from inlet data obtained during steady aerodynamic maneuvers. Results show that rapid-maneuver inlet recoveries agreed very well with the recoveries obtained at equivalent stabilized angle-of-attack conditions. The peak dynamic circumferential distortion values obtained during rapid maneuvers agreed within 0.01 units of distortion over the 10 - 38 degree angle of attack range with the values obtained during steady aerodynamic maneuvers while similar agreement was found for the peak dynamic radial distortion values up to 29 degrees angle-of-attack. Exceedences of the rapid-maneuver peak dynamic circumferential distortion values relative to the peak distortion model values at steady attitudes occurred only at low or negative angles of attack and were inconsequential from an engine-stability assessment point of view. The results of this study validate the current industry practice of testing at steady aerodynamic conditions to characterize inlet recovery and peak dynamic distortion levels.

  18. A Phasin with Many Faces: Structural Insights on PhaP from Azotobacter sp. FA8

    PubMed Central

    Mezzina, Mariela P.; Wetzler, Diana E.; Catone, Mariela V.; Bucci, Hernan; Di Paola, Matias; Pettinari, M. Julia

    2014-01-01

    Phasins are a group of proteins associated to granules of polyhydroxyalkanoates (PHAs). Apart from their structural role as part of the PHA granule cover, different structural and regulatory functions have been found associated to many of them, and several biotechnological applications have been developed using phasin protein fusions. Despite their remarkable functional diversity, the structure of these proteins has not been analyzed except in very few studies. PhaP from Azotobacter sp. FA8 (PhaPAz) is a representative of the prevailing type in the multifunctional phasin protein family. Previous work performed in our laboratory using this protein have demonstrated that it has some very peculiar characteristics, such as its stress protecting effects in recombinant Escherichia coli, both in the presence and absence of PHA. The aim of the present work was to perform a structural characterization of this protein, to shed light on its properties. Its aminoacid composition revealed that it lacks clear hydrophobic domains, a characteristic that appears to be common to most phasins, despite their lipid granule binding capacity. The secondary structure of this protein, consisting of α-helices and disordered regions, has a remarkable capacity to change according to its environment. Several experimental data support that it is a tetramer, probably due to interactions between coiled-coil regions. These structural features have also been detected in other phasins, and may be related to their functional diversity. PMID:25077609

  19. Spatial Characteristics of the Unsteady Differential Pressures on 16 percent F/A-18 Vertical Tails

    NASA Technical Reports Server (NTRS)

    Moses, Robert W.; Ashley, Holt

    1998-01-01

    Buffeting is an aeroelastic phenomenon which plagues high performance aircraft at high angles of attack. For the F/A-18 at high angles of attack, vortices emanating from wing/fuselage leading edge extensions burst, immersing the vertical tails in their turbulent wake. The resulting buffeting of the vertical tails is a concern from fatigue and inspection points of view. Previous flight and wind-tunnel investigations to determine the buffet loads on the tail did not provide a complete description of the spatial characteristics of the unsteady differential pressures. Consequently, the unsteady differential pressures were considered to be fully correlated in the analyses of buffet and buffeting. The use of fully correlated pressures in estimating the generalized aerodynamic forces for the analysis of buffeting yielded responses that exceeded those measured in flight and in the wind tunnel. To learn more about the spatial characteristics of the unsteady differential pressures, an available 16%, sting-mounted, F-18 wind-tunnel model was modified and tested in the Transonic Dynamics Tunnel (TDT) at the NASA Langley Research Center as part of the ACROBAT (Actively Controlled Response Of Buffet-Affected Tails) program. Surface pressures were measured at high angles of attack on flexible and rigid tails. Cross-correlation and cross-spectral analyses of the pressure time histories indicate that the unsteady differential pressures are not fully correlated. In fact, the unsteady differential pressure resemble a wave that travels along the tail. At constant angle of attack, the pressure correlation varies with flight speed.

  20. Summary of Inlet Characteristics of the F/A-18A High Alpha Research Vehicle

    NASA Technical Reports Server (NTRS)

    Walsh, Kevin; Steenken, William; Williams, John

    1998-01-01

    Effects of high-angle-of-attack flight on aircraft inlet aerodynamic characteristics were investigated at NASA Dryden Flight Research Center as part of NASA's High Alpha Technology Program. The highly instrumented F/A-18A High Alpha Research Vehicle was used for this research. A newly designed inlet total-pressure rake was installed in front of the right-hand F404-GE-400 engine to measure inlet recovery and distortion characteristics. Objectives included: (1) determining the inlet total-pressure characteristics at steady high-angle-of-attack conditions; (2) assessing if inlet distortion is significantly different between rapid angle-of-attack maneuvers and corresponding steady aerodynamic conditions; (3) assessing inlet characteristics during aircraft departures; (4) providing data for developing and verifying computational fluid dynamic codes; and (5) calculating engine airflow using four methods for comparison with a reference method. This paper describes the results obtained from this investigation. These data and the associated database were rigorously validated to establish the foundation for understanding inlet characteristics at high angle of attack.

  1. Aeroservoelastic Model Validation and Test Data Analysis of the F/A-18 Active Aeroelastic Wing

    NASA Technical Reports Server (NTRS)

    Brenner, Martin J.; Prazenica, Richard J.

    2003-01-01

    Model validation and flight test data analysis require careful consideration of the effects of uncertainty, noise, and nonlinearity. Uncertainty prevails in the data analysis techniques and results in a composite model uncertainty from unmodeled dynamics, assumptions and mechanics of the estimation procedures, noise, and nonlinearity. A fundamental requirement for reliable and robust model development is an attempt to account for each of these sources of error, in particular, for model validation, robust stability prediction, and flight control system development. This paper is concerned with data processing procedures for uncertainty reduction in model validation for stability estimation and nonlinear identification. F/A-18 Active Aeroelastic Wing (AAW) aircraft data is used to demonstrate signal representation effects on uncertain model development, stability estimation, and nonlinear identification. Data is decomposed using adaptive orthonormal best-basis and wavelet-basis signal decompositions for signal denoising into linear and nonlinear identification algorithms. Nonlinear identification from a wavelet-based Volterra kernel procedure is used to extract nonlinear dynamics from aeroelastic responses, and to assist model development and uncertainty reduction for model validation and stability prediction by removing a class of nonlinearity from the uncertainty.

  2. Starmerella orientalis f.a., sp. nov., an ascomycetous yeast species isolated from flowers.

    PubMed

    Alimadadi, Nayyereh; Soudi, Mohammad Reza; Wang, Shi-An; Wang, Qi-Ming; Talebpour, Zahra; Bai, Feng-Yan

    2016-03-01

    Four strains of a novel ascomycetous yeast species were isolated from flowers in Iran and China. Phylogenetic analysis of the sequences of the ITS region (including 5.8S rRNA gene) and the LSU rRNA gene D1/D2 domains indicated that these strains belong to the Starmerella clade and show divergence from previously described species in this clade. Growth reactions on carbon and nitrogen sources were similar to those observed in related species of the Starmerella clade. Sexual reproduction was not observed after mating tests on different sporulation media. Based on physiological characteristics and phylogeny of rRNA gene sequences, the novel species is most closely related to Candida (iter. nom. Starmerella) powellii and Candida (iter. nom. Starmerella) floricola. It is therefore assigned to the genus Starmerella and described as Starmerella orientalis f.a., sp. nov. The type strain is SAM09T ( = IBRC-M 30204T = CBS 14142T). The MycoBank accession number is MB 814379. PMID:26780917

  3. Deflection-Based Structural Loads Estimation From the Active Aeroelastic Wing F/A-18 Aircraft

    NASA Technical Reports Server (NTRS)

    Lizotte, Andrew M.; Lokos, William A.

    2005-01-01

    Traditional techniques in structural load measurement entail the correlation of a known load with strain-gage output from the individual components of a structure or machine. The use of strain gages has proved successful and is considered the standard approach for load measurement. However, remotely measuring aerodynamic loads using deflection measurement systems to determine aeroelastic deformation as a substitute to strain gages may yield lower testing costs while improving aircraft performance through reduced instrumentation weight. This technique was examined using a reliable strain and structural deformation measurement system. The objective of this study was to explore the utility of a deflection-based load estimation, using the active aeroelastic wing F/A-18 aircraft. Calibration data from ground tests performed on the aircraft were used to derive left wing-root and wing-fold bending-moment and torque load equations based on strain gages, however, for this study, point deflections were used to derive deflection-based load equations. Comparisons between the strain-gage and deflection-based methods are presented. Flight data from the phase-1 active aeroelastic wing flight program were used to validate the deflection-based load estimation method. Flight validation revealed a strong bending-moment correlation and slightly weaker torque correlation. Development of current techniques, and future studies are discussed.

  4. Papiliotrema siamense f.a., sp. nov., a yeast species isolated from plant leaves.

    PubMed

    Surussawadee, Janjira; Khunnamwong, Pannida; Srisuk, Nantana; Limtong, Savitree

    2014-09-01

    Two strains representing a novel species were isolated from the external surface of a sugar cane leaf (DMKU-SP85(T)) and tissue of a rice leaf (DMKU-RE97) collected in Thailand. On the basis of morphological, biochemical, physiological and chemotaxonomic characteristics, and sequence analysis of the D1/D2 region of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) region, the two strains were determined to represent a novel species of the genus Papiliotrema although sexual reproduction was not observed. The sequences of the D1/D2 region of the LSU rRNA gene and ITS region of the two strains were identical, but differed from those of the type strain of Cryptococcus nemorosus by 0.6 % nucleotide substitutions (four nucleotide substitutions out of 597 nucleotides) in the D1/D2 region of the LSU rRNA gene and 1.8 % nucleotide substitutions (nine nucleotide substitutions out of 499 nucleotides) in the ITS region. The name Papiliotrema siamense f.a., sp. nov. is proposed. The type strain is DMKU-SP85(T)( = BCC 69499(T) = CBS 13330(T)). PMID:24925598

  5. The aerodynamic characteristics of vortex ingestion for the F/A-18 inlet duct

    NASA Technical Reports Server (NTRS)

    Anderson, Bernhard H.

    1991-01-01

    A Reduced Navier-Stokes (RNS) solution technique was successfully combined with the concept of partitioned geometry and mesh generation to form a very efficient 3D RNS code aimed at the analysis-design engineering environment. Partitioned geometry and mesh generation is a pre-processor to augment existing geometry and grid generation programs which allows the solver to (1) recluster an existing gridlife mesh lattice, and (2) perturb an existing gridfile definition to alter the cross-sectional shape and inlet duct centerline distribution without returning to the external geometry and grid generator. The present results provide a quantitative validation of the initial value space marching 3D RNS procedure and demonstrates accurate predictions of the engine face flow field, with a separation present in the inlet duct as well as when vortex generators are installed to supress flow separation. The present results also demonstrate the ability of the 3D RNS procedure to analyze the flow physics associated with vortex ingestion in general geometry ducts such as the F/A-18 inlet. At the conditions investigated, these interactions are basically inviscid like, i.e., the dominant aerodynamic characteristics have their origin in inviscid flow theory.

  6. Strain Gage Loads Calibration Testing of the Active Aeroelastic Wing F/A-18 Aircraft

    NASA Technical Reports Server (NTRS)

    Lokos, William A.; Olney, Candida D.; Chen, Tony; Crawford, Natalie D.; Stauf, Rick; Reichenbach, Eric Y.; Bessette, Denis (Technical Monitor)

    2002-01-01

    This report describes strain-gage calibration loading through the application of known loads of the Active Aeroelastic Wing F/A-18 airplane. The primary goal of this test is to produce a database suitable for deriving load equations for left and right wing root and fold shear; bending moment; torque; and all eight wing control-surface hinge moments. A secondary goal is to produce a database of wing deflections measured by string potentiometers and the onboard flight deflection measurement system. Another goal is to produce strain-gage data through both the laboratory data acquisition system and the onboard aircraft data system as a check of the aircraft system. Thirty-two hydraulic jacks have applied loads through whiffletrees to 104 tension-compression load pads bonded to the lower wing surfaces. The load pads covered approximately 60 percent of the lower wing surface. A series of 72 load cases has been performed, including single-point, double-point, and distributed load cases. Applied loads have reached 70 percent of the flight limit load. Maximum wingtip deflection has reached nearly 16 in.

  7. Acoustic emission monitoring of a fatigue test of an F/A-18 bulkhead

    NASA Astrophysics Data System (ADS)

    Scala, C. M.; McCardle, J. F.; Bowles, S. J.

    This paper describes the application of acoustic emission (AE) to identify cracking in several fatigue-critical regions on the port and starboard sides of an l/A-18 aircraft bulkhead undergoing fatigue testing. AE data acquisition was carried out using an array of three sensors on each side of the bulkhead. AE features stored by each array included relative arrival times of AE events at the three sensors, event rise time at the first-hit sensor, and the load level and the position on the load cycle of event occurrence. AE data processing involved a comparison between the features of those AE events stored during the fatigue testing and predicted features for cracking in the complex-shaped bulkhead. Feature prediction was based on wave propagation characteristics obtained by Pentel-lead calibration, and the known load cycle dependence of crack-related AE events. The AE processing was completed following failure of the bulkhead, and gave the correct locations of all cracks, greater than about 1 mm in depth, present in the bulkhead during the fatigue testing. The study shows that AE associated with cracking can be distinguished, even when many extraneous sources are present, and demonstrates that AE is a promising technique for nondestructive evaluation of a complex structure such as the F/A-18 bulkhead.

  8. Acoustic emission testing on an F/A-18 E/F titanium bulkhead

    NASA Astrophysics Data System (ADS)

    Martin, Christopher A.; Van Way, Craig B.; Lockyer, Allen J.; Kudva, Jayanth N.; Ziola, Steve M.

    1995-04-01

    An important opportunity recently transpired at Northrop Grumman Corporation to instrument an F/A - 18 E/F titanium bulkhead with broad band acoustic emission sensors during a scheduled structural fatigue test. The overall intention of this effort was to investigate the potential for detecting crack propagation using acoustic transmission signals for a large structural component. Key areas of experimentation and experience included (1) acoustic noise characterization, (2) separation of crack signals from extraneous noise, (3) source location accuracy, and (4) methods of acoustic transducer attachment. Fatigue cracking was observed and monitored by strategically placed acoustic emission sensors. The outcome of the testing indicated that accurate source location still remains enigmatic for non-specialist engineering personnel especially at this level of structural complexity. However, contrary to preconceived expectations, crack events could be readily separated from extraneous noise. A further dividend from the investigation materialized in the form of close correspondence between frequency domain waveforms of the bulkhead test specimen tested and earlier work with thick plates.

  9. Induced Moment Effects of Formation Flight Using Two F/A-18 Aircraft

    NASA Technical Reports Server (NTRS)

    Hansen, Jennifer L.; Cobleigh, Brent R.

    2002-01-01

    Previous investigations into formation flight have shown the possibility for significant fuel savings through drag reduction. Using two F/A-18 aircraft, NASA Dryden Flight Research Center has investigated flying aircraft in autonomous formation. Positioning the trailing airplane for best drag reduction requires investigation of the wingtip vortex effects induced by the leading airplane. A full accounting of the vortex effect on the trailing airplane is desired to validate vortex-effect prediction methods and provide a database for the design of a formation flight autopilot. A recent flight phase has mapped the complete wingtip vortex effects at two flight conditions with the trailing airplane at varying distances behind the leading one. Force and moment data at Mach 0.56 and an altitude of 25,000 ft and Mach 0.86 and an altitude of 36,000 ft have been obtained with 20, 55, 110, and 190 ft of longitudinal distance between the aircraft. The moments induced by the vortex on the trailing airplane were well within the pilot's ability to control. This report discusses the data analysis methods and vortex-induced effects on moments and side force. An assessment of the impact of the nonlinear vortex effects on the design of a formation autopilot is offered.

  10. Twist Model Development and Results From the Active Aeroelastic Wing F/A-18 Aircraft

    NASA Technical Reports Server (NTRS)

    Lizotte, Andrew; Allen, Michael J.

    2005-01-01

    Understanding the wing twist of the active aeroelastic wing F/A-18 aircraft is a fundamental research objective for the program and offers numerous benefits. In order to clearly understand the wing flexibility characteristics, a model was created to predict real-time wing twist. A reliable twist model allows the prediction of twist for flight simulation, provides insight into aircraft performance uncertainties, and assists with computational fluid dynamic and aeroelastic issues. The left wing of the aircraft was heavily instrumented during the first phase of the active aeroelastic wing program allowing deflection data collection. Traditional data processing steps were taken to reduce flight data, and twist predictions were made using linear regression techniques. The model predictions determined a consistent linear relationship between the measured twist and aircraft parameters, such as surface positions and aircraft state variables. Error in the original model was reduced in some cases by using a dynamic pressure-based assumption and by using neural networks. These techniques produced excellent predictions for flight between the standard test points and accounted for nonlinearities in the data. This report discusses data processing techniques and twist prediction validation, and provides illustrative and quantitative results.

  11. Twist Model Development and Results from the Active Aeroelastic Wing F/A-18 Aircraft

    NASA Technical Reports Server (NTRS)

    Lizotte, Andrew M.; Allen, Michael J.

    2007-01-01

    Understanding the wing twist of the active aeroelastic wing (AAW) F/A-18 aircraft is a fundamental research objective for the program and offers numerous benefits. In order to clearly understand the wing flexibility characteristics, a model was created to predict real-time wing twist. A reliable twist model allows the prediction of twist for flight simulation, provides insight into aircraft performance uncertainties, and assists with computational fluid dynamic and aeroelastic issues. The left wing of the aircraft was heavily instrumented during the first phase of the active aeroelastic wing program allowing deflection data collection. Traditional data processing steps were taken to reduce flight data, and twist predictions were made using linear regression techniques. The model predictions determined a consistent linear relationship between the measured twist and aircraft parameters, such as surface positions and aircraft state variables. Error in the original model was reduced in some cases by using a dynamic pressure-based assumption. This technique produced excellent predictions for flight between the standard test points and accounted for nonlinearities in the data. This report discusses data processing techniques and twist prediction validation, and provides illustrative and quantitative results.

  12. Review of Full-Scale F/A-18 Research at NASA Ames Research Center

    NASA Technical Reports Server (NTRS)

    Olson, Lawrence E.; Schmitz, Fredric H. (Technical Monitor)

    1994-01-01

    Results of flow visualization and tail buffett studies conducted on a full-scale production F/A-18 fighter aircraft in the 80- by 120-Foot Wind Tunnel of the National Full-Scale Aerodynamic Complex are presented. Test conditions range between 20 degrees and 40 degrees angle of attack, 16 degrees and -16 degrees side-slip angle, and up to a Mach number of 0.15 (corresponding to a Reynolds number of 12.3 x 10(exp 6) based on mean aerodynamic chord). Flow visualization results include both surface and off-surface techniques that examine forebody, canopy, leading-edge extension, and wing flow fields. Unsteady pressures measured at 96 locations on the port tail fin are used to determine the effect of a removable leading-edge extension fence on tail buffet loads at high angle of attack. Analyses and comparisons include tail fin bending moment and wave velocities on the tail surface. Repeatability and scaling issues are assessed through comparison with measurements from previous full-scale tests and several small-scales tests.

  13. Functional characterization of FaNIP1;1 gene, a ripening-related and receptacle-specific aquaporin in strawberry fruit.

    PubMed

    Molina-Hidalgo, Francisco J; Medina-Puche, Laura; Gelis, Samuel; Ramos, José; Sabir, Farzana; Soveral, Graça; Prista, Catarina; Iglesias-Fernández, Raquel; Caballero, José L; Muñoz-Blanco, Juan; Blanco-Portales, Rosario

    2015-09-01

    Strawberry fruit (Fragaria × ananassa) is a soft fruit with high water content at ripe stage (more than 90% of its fresh weight). Aquaporins play an important role in plant water homeostasis, through the facilitation of water transport and solutes. We report the role played by FaNIP1;1 in the receptacle ripening process. The analysis by qRT-PCR of FaNIP1;1 showed that this gene is mainly expressed in fruit receptacle and has a ripening-related expression pattern that was accompanied by an increase in both the abscisic acid and water content of the receptacle throughout fruit ripening. Moreover, FaNIP1;1 was induced in situations of water deficit. Additionally, we show that FaNIP1;1 expression was positively regulated by abscisic acid and negatively regulated by auxins. The water transport capacity of FaNIP1;1 was determined by a stopped-flow spectroscopy in yeast over-expressing FaNIP1;1. Glycerol, H2O2 and boron transport were also demonstrated in yeast. On the other hand, GFP-FaNIP1;1 fusion protein was located in plasma membrane. In conclusion, FaNIP1;1 seems to play an important role increasing the plasma membrane permeability, that allows the water accumulation in the strawberry fruit receptacle throughout the ripening process. PMID:26259188

  14. Suppression of Human T Cell Proliferation Mediated by the Cathepsin B Inhibitor, z-FA-FMK Is Due to Oxidative Stress

    PubMed Central

    Rajah, Tanuja; Chow, Sek Chuen

    2015-01-01

    The cathepsin B inhibitor, benzyloxycarbonyl-phenylalanine-alanine-fluoromethyl ketone (z-FA-FMK) readily inhibits anti-CD3-induced human T cell proliferation, whereas the analogue benzyloxycarbonyl-phenylalanine-alanine-diazomethyl ketone (z-FA-DMK) had no effect. In contrast, benzyloxycarbonyl-phenylalanine-alanine-chloromethyl ketone (z-FA-CMK) was toxic. The inhibition of T cell proliferation mediated by z-FA-FMK requires not only the FMK moiety, but also the benzyloxycarbonyl group at the N-terminal, suggesting some degree of specificity in z-FA-FMK-induced inhibition of primary T cell proliferation. We showed that z-FA-FMK treatment leads to a decrease in intracellular glutathione (GSH) with a concomitant increase in reactive oxygen species (ROS) levels in activated T cells. The inhibition of anti-CD3-induced T cell proliferation mediated by z-FA-FMK was abolished by the presence of low molecular weight thiols such as GSH, N-acetylcysteine (NAC) and L-cysteine, whereas D-cysteine which cannot be metabolised to GSH has no effect. The inhibition of anti-CD3-induced up-regulation of CD25 and CD69 expression mediated by z-FA-FMK was also attenuated in the presence of exogenous GSH. Similar to cell proliferation, GSH, NAC and L-cysteine but not D-cysteine, completely restored the processing of caspase-8 and caspase-3 to their respective subunits in z-FA-FMK-treated activated T cells. Our collective results demonstrated that the inhibition of T cell activation and proliferation mediated by z-FA-FMK is due to oxidative stress via the depletion of GSH. PMID:25915766

  15. Organometallic Complex Formed by an Unconventional Radical S-Adenosylmethionine Enzyme.

    PubMed

    Dong, Min; Horitani, Masaki; Dzikovski, Boris; Pandelia, Maria-Eirini; Krebs, Carsten; Freed, Jack H; Hoffman, Brian M; Lin, Hening

    2016-08-10

    Pyrococcus horikoshii Dph2 (PhDph2) is an unusual radical S-adenosylmethionine (SAM) enzyme involved in the first step of diphthamide biosynthesis. It catalyzes the reaction by cleaving SAM to generate a 3-amino-3-carboxypropyl (ACP) radical. To probe the reaction mechanism, we synthesized a SAM analogue (SAMCA), in which the ACP group of SAM is replaced with a 3-carboxyallyl group. SAMCA is cleaved by PhDph2, yielding a paramagnetic (S = 1/2) species, which is assigned to a complex formed between the reaction product, α-sulfinyl-3-butenoic acid, and the [4Fe-4S] cluster. Electron-nuclear double resonance (ENDOR) measurements with (13)C and (2)H isotopically labeled SAMCA support a π-complex between the C═C double bond of α-sulfinyl-3-butenoic acid and the unique iron of the [4Fe-4S] cluster. This is the first example of a radical SAM-related [4Fe-4S](+) cluster forming an organometallic complex with an alkene, shedding additional light on the mechanism of PhDph2 and expanding our current notions for the reactivity of [4Fe-4S] clusters in radical SAM enzymes. PMID:27465315

  16. GLUCOMANNAN AND GLUCOMANNAN PLUS SPIRULINA-ENRICHED SQUID-SURIMI ADDED TO HIGH SATURATED DIET AFFECT GLYCEMIA, PLASMA AND ADIPOSE LEPTIN AND ADIPONECTIN LEVELS IN GROWING FA/FA RATS.

    PubMed

    Vázquez-Velasco, Miguel; González-Torres, Laura; Méndez, María Teresa; Bastida, Sara; Benedí, Juana; González-Muñoz, M José; Sánchez-Muniz, Francisco J

    2015-01-01

    Type 2 diabetes is a very prevalent chronic disease. Among dietary factors for its prevention and treatment, interest has grown in satiating fibre (konjac glucomannan) and spirulina. Our previous studies suggest that glucomannan itself and/or in conjunction to spirulina displayed hypolipemic and antioxidant effects when incorporated to squid surimi as functional ingredients. The present study aims to determine whether glucomannan- enriched or glucomannan plus spirulina-enriched squid-surimi improve plasma glucose and insulin levels in Zucker fa/fa rats fed a high saturated fat diet. Twenty four growing rats, divided into three groups, were given modified AIN-93M diets for seven weeks: 30% squid-surimi control diet (C), 30% glucomannan-enriched squid-surimi diet (G) and 30% glucomannan plus spirulina-enriched squid-surimi diet (GS). All rats became hyperglycemics and hyperinsulinemics, but G and GS diets induced significantly lower glucose levels (20%; p < 0.05) but did not modify insulinemia with respect to C diet. GS animals showed higher HOMA-D (p < 0.05) than C ones suggesting increased insulin availability. Plasma leptin and adiponectin decreased in G and GS vs. C group (p < 0.05). Adipose adiponectin increased significantly in G and GS vs. C rats (16-20 times, p < 0.01). Leptin in adipose tissue was higher in GS vs. G group (p < 0.05). In conclusion, both glucomannan-diets were able to reduce hyperglycemia and increase adipose tissue adiponectin levels in fa/fa rats, suggesting an anti-hypertrophic and insulin-sensitizing adipokine effect in this tissue. Spirulina inclusion increased insulin availability. Although results are promising, the utility of consuming glucomannan surimis as part of usual diets demands future studies. PMID:26667726

  17. Navier-Stokes, flight, and wind tunnel flow analysis for the F/A-18 aircraft

    NASA Technical Reports Server (NTRS)

    Ghaffari, Farhad

    1994-01-01

    Computational analysis of flow over the F/A-18 aircraft is presented along with complementary data from both flight and wind tunnel experiments. The computational results are based on the three-dimensional thin-layer Navier-Stokes formulation and are obtained from an accurate surface representation of the fuselage, leading-edge extension (LEX), and the wing geometry. However, the constraints imposed by either the flow solver and/or the complexity associated with the flow-field grid generation required certain geometrical approximations to be implemented in the present numerical model. In particular, such constraints inspired the removal of the empennage and the blocking (fairing) of the inlet face. The results are computed for three different free-stream flow conditions and compared with flight test data of surface pressure coefficients, surface tuft flow, and off-surface vortical flow characteristics that included breakdown phenomena. Excellent surface pressure coefficient correlations, both in terms of magnitude and overall trend, are obtained on the forebody throughout the range of flow conditions. Reasonable pressure agreement was obtained over the LEX; the general correlation tends to improve at higher angles of attack. The surface tuft flow and the off-surface vortex flow structures compared qualitatively well with the flight test results. To evaluate the computational results, a wind tunnel investigation was conducted to determine the effects of existing configurational differences between the flight vehicle and the numerical model on aerodynamic characteristics. In most cases, the geometrical approximations made to the numerical model had very little effect on overall aerodynamic characteristics.

  18. Transonic Unsteady Aerodynamics of the F/A-18E at Conditions Promoting Abrupt Wing Stall

    NASA Technical Reports Server (NTRS)

    Schuster, David M.; Byrd, James E.

    2003-01-01

    A transonic wind tunnel test of an 8% F/A-18E model was conducted in the NASA Langley Research Center (LaRC) 16-Foot Transonic Tunnel (16-Ft TT) to investigate the Abrupt Wing Stall (AWS) characteristics of this aircraft. During this test, both steady and unsteady measurements of balance loads, wing surface pressures, wing root bending moments, and outer wing accelerations were performed. The test was conducted with a wide range of model configurations and test conditions in an attempt to reproduce behavior indicative of the AWS phenomenon experienced on full-scale aircraft during flight tests. This paper focuses on the analysis of the unsteady data acquired during this test. Though the test apparatus was designed to be effectively rigid. model motions due to sting and balance flexibility were observed during the testing, particularly when the model was operating in the AWS flight regime. Correlation between observed aerodynamic frequencies and model structural frequencies are analyzed and presented. Significant shock motion and separated flow is observed as the aircraft pitches through the AWS region. A shock tracking strategy has been formulated to observe this phenomenon. Using this technique, the range of shock motion is readily determined as the aircraft encounters AWS conditions. Spectral analysis of the shock motion shows the frequencies at which the shock oscillates in the AWS region, and probability density function analysis of the shock location shows the propensity of the shock to take on a bi-stable and even tri-stable character in the AWS flight regime.

  19. Design and development of an F/A-18 inlet distortion rake: A cost and time saving solution

    NASA Technical Reports Server (NTRS)

    Yuhas, Andrew J.; Ray, Ronald J.; Burley, Richard R.; Steenken, William G.; Lechtenberg, Leon; Thornton, Don

    1995-01-01

    An innovative inlet total pressure distortion measurement rake has been designed and developed for the F/A-18 A/B/C/D aircraft inlet. The design was conceived by NASA and General Electric Aircraft Engines personnel. This rake has been flight qualified and flown in the F/A-18 High Alpha Research Vehicle at NASA Dryden Flight Research Center, Edwards, California. The eight-legged, one-piece, wagon wheel design of the rake was developed at a reduced cost and offered reduced installation time compared to traditional designs. The rake features 40 dual-measurement ports for low- and high-frequency pressure measurements with the high-frequency transducer mounted at the port. This high-frequency transducer offers direct absolute pressure measurements from low to high frequencies of interest, thereby allowing the rake to be used during highly dynamic aircraft maneuvers. Outstanding structural characteristics are inherent to the design through its construction and use of lightweight materials.

  20. Birth order in transgendered males from Polynesia: a quantitative study of Samoan fa'afāfine.

    PubMed

    Poasa, Kris H; Blanchard, Ray; Zucker, Kenneth J

    2004-01-01

    In previous research, a late birth order has been shown to be a characteristic of Western homosexual transsexual men. To date, however, it is not clear if a late birth order is specific to Western transsexuals or may be a characteristic of non-Western transgendered males as well. We quantified birth order in a sample of 13 transgendered males known as fa'afāfine from Samoa. On average, the fa'afāfine were late born, both with regard to number of older brothers and number of older sisters, although the effect appeared to be somewhat stronger with regard to number of older brothers. We consider possible interpretations of the late birth order effect, along with recommendations for further research to identify common mechanisms that might underlie transgenderism across different cultural groups. PMID:14660290

  1. Marbles in the façade of the "Certosa di Pavia": a physico-chemical study.

    PubMed

    Ferloni, Paolo; Chierichetti, Andrea; Tomasi, Corrado; Ricci, Oronzo

    2005-01-01

    In the framework of an interdisciplinary research project on the well known monastery "Certosa di Pavia", the thermal and structural properties of marbles employed in the construction of the façade of the basilica were investigated in order to detect the main decomposition phenomena occurring on the monument surface. The results obtained by means of thermogravimetric and differential thermal analyses, as well as by means of X-ray diffraction and IR spectroscopy, allowed one to characterize samples taken from various sculptures of the façade and to bring out the degradation phenomena occurring in some of them, in particular the formation of "black crusts" with a high content in gypsum. The present findings are in fair agreement with those reported by other research groups which also studied with different techniques the construction materials of this monument. PMID:16485656

  2. Navier-Stokes solutions about the F/A-18 forebody-LEX configuration. [Leading Edge Extension

    NASA Technical Reports Server (NTRS)

    Ghaffari, Farhad; Bates, Brent L.; Luckring, James M.; Thomas, James L.

    1989-01-01

    Three-dimensional viscous flow computations are presented for the F/A-18 forebody-LEX geometry. Solutions are obtained from an algorithm for the compressible Navier-Stokes equations which incorporates an upwind-biased, flux-difference-splitting approach along with longitudinally-patched grids. Results are presented for both laminar and fully turbulent flow assumptions and include correlations with wind tunnel as well as flight-test results. A good quantitative agreement for the forebody surface pressure distribution is achieved between the turbulent computations and wind tunnel measurements at a free-stream Mach number of 0.6. The computed turbulent surface flow patterns on the forebody qualitatively agree well with in-flight surface flow patterns obtained on an F/A-18 aircraft at a free-stream Mach number of 0.34.

  3. A NASA F/A-18, participating in the Automated Aerial Refueling (AAR) project, flies over the Dryden

    NASA Technical Reports Server (NTRS)

    2002-01-01

    A NASA F/A-18 flies over the Dryden Flight Research Center and Rogers Dry Lake on December 11, 2002. The aircraft is participating in the Automated Aerial Refueling (AAR) project. The 300-gallon aerial refueling store seen on the belly of the aircraft carries fuel and a refueling drogue. This aircraft acts as a tanker in the study to develop an aerodynamic model for future automated aerial refueling, especially of unmanned vehicles.

  4. An H-Infinity Approach to Control Synthesis with Load Minimization for the F/A-18 Active Aeroelastic Wing

    NASA Technical Reports Server (NTRS)

    Lind, Rick

    1999-01-01

    The F/A-18 Active Aeroelastic Wing research aircraft will demonstrate technologies related to aeroservoelastic effects such as wing twist and load minimization. This program presents several challenges for control design that are often not considered for traditional aircraft. This paper presents a control design based on H-infinity synthesis that simultaneously considers the multiple objectives associated with handling qualities, actuator limitations, and loads. A point design is presented to demonstrate a controller and the resulting closed-loop properties.

  5. An H-infinity Approach to Control Synthesis with Load Minimization for the F/A-18 Active Aeroelastic Wing

    NASA Technical Reports Server (NTRS)

    Lind, Rick

    1999-01-01

    The F/A-18 Active Aeroelastic Wing research aircraft will demonstrate technologies related to aeroservoelastic effects such as wing twist and load minimization. This program presents several challenges for control design that are often not considered for traditional aircraft. This paper presents a control design based on H(sub infinity) synthesis that simultaneously considers the multiple objectives associated with handling qualities, actuator limitations, and loads. A point design is presented to demonstrate a controller and the resulting closed-loop properties.

  6. A water tunnel flow visualization study of the vortex flow structures on the F/A-18 aircraft

    NASA Technical Reports Server (NTRS)

    Sandlin, Doral R.; Ramirez, Edgar J.

    1991-01-01

    The vortex flow structures occurring on the F/A-18 aircraft at high angles of attack were studied. A water tunnel was used to gather flow visualization data on the forebody vortex and the wing leading edge extension vortex. The longitudinal location of breakdown of the leading edge vortex was found to be consistently dependent on the angle of attack. Other parameters such as Reynolds number, model scale, and model fidelity had little influence on the overall behavior of the flow structures studied. The lateral location of the forebody vortex system was greatly influenced by changes in the angle of sideslip. Strong interactions can occur between the leading edge extension vortex and the forebody vortex. Close attention was paid to vortex induced flows on various airframe components of the F/A-18. Reynolds number and angle of attack greatly affected the swirling intensity, and therefore the strength of the studied vortices. Water tunnel results on the F/A-18 correlated well with those obtained in similar studies at both full and sub scale levels. The water tunnel can provide, under certain conditions, good simulations of realistic flows in full scale configurations.

  7. Fluorophore Absorption Size Exclusion Chromatography (FA-SEC): An Alternative Method for High-Throughput Detergent Screening of Membrane Proteins.

    PubMed

    Lin, Sung-Yao; Sun, Xing-Han; Hsiao, Yu-Hsuan; Chang, Shao-En; Li, Guan-Syun; Hu, Nien-Jen

    2016-01-01

    Membrane proteins play key roles in many fundamental functions in cells including ATP synthesis, ion and molecule transporter, cell signalling and enzymatic reactions, accounting for ~30% genes of whole genomes. However, the hydrophobic nature of membrane proteins frequently hampers the progress of structure determination. Detergent screening is the critical step in obtaining stable detergent-solubilized membrane proteins and well-diffracting protein crystals. Fluorescence Detection Size Exclusion Chromatography (FSEC) has been developed to monitor the extraction efficiency and monodispersity of membrane proteins in detergent micelles. By tracing the FSEC profiles of GFP-fused membrane proteins, this method significantly enhances the throughput of detergent screening. However, current methods to acquire FSEC profiles require either an in-line fluorescence detector with the SEC equipment or an off-line spectrofluorometer microplate reader. Here, we introduce an alternative method detecting the absorption of GFP (FA-SEC) at 485 nm, thus making this methodology possible on conventional SEC equipment through the in-line absorbance spectrometer. The results demonstrate that absorption is in great correlation with fluorescence of GFP. The comparably weaker absorption signal can be improved by using a longer path-length flow cell. The FA-SEC profiles were congruent with the ones plotted by FSEC, suggesting FA-SEC could be a comparable and economical setup for detergent screening of membrane proteins. PMID:27332877

  8. Free-to-Roll Investigation of the Pre-Production F/A-18E Powered Approach Wing Drop

    NASA Technical Reports Server (NTRS)

    Owens, D. Bruce; Bryant, Elaine M.; Barlow, Jewel B.

    2005-01-01

    A free-to-roll study of the low-speed lateral characteristics of the pre-production F/A-18E was conducted in the NASA Langley 12-Foot Low-Speed Tunnel. In developmental flight tests the F/A-18E unexpectedly experienced uncommanded lateral motions in the power approach configuration. The objective of this study was to determine the feasibility of using the free-to-roll technique for the detection of uncommanded lateral motions for the pre-production F/A-18E in the power approach configuration. The data revealed that this technique in conjunction with static data revealed insight into the cause of the lateral motions. The free-to-roll technique identified uncommanded lateral motions at the same angle-of-attack range as experienced in flight tests. The cause of the uncommanded lateral motions was unsteady asymmetric wing stall. The paper also shows that free-to-roll data or static force and moment data alone are not enough to accurately capture the potential for an aircraft to experience uncommanded lateral motion.

  9. Fluorophore Absorption Size Exclusion Chromatography (FA-SEC): An Alternative Method for High-Throughput Detergent Screening of Membrane Proteins

    PubMed Central

    Hsiao, Yu-Hsuan; Chang, Shao-En; Li, Guan-Syun; Hu, Nien-Jen

    2016-01-01

    Membrane proteins play key roles in many fundamental functions in cells including ATP synthesis, ion and molecule transporter, cell signalling and enzymatic reactions, accounting for ~30% genes of whole genomes. However, the hydrophobic nature of membrane proteins frequently hampers the progress of structure determination. Detergent screening is the critical step in obtaining stable detergent-solubilized membrane proteins and well-diffracting protein crystals. Fluorescence Detection Size Exclusion Chromatography (FSEC) has been developed to monitor the extraction efficiency and monodispersity of membrane proteins in detergent micelles. By tracing the FSEC profiles of GFP-fused membrane proteins, this method significantly enhances the throughput of detergent screening. However, current methods to acquire FSEC profiles require either an in-line fluorescence detector with the SEC equipment or an off-line spectrofluorometer microplate reader. Here, we introduce an alternative method detecting the absorption of GFP (FA-SEC) at 485 nm, thus making this methodology possible on conventional SEC equipment through the in-line absorbance spectrometer. The results demonstrate that absorption is in great correlation with fluorescence of GFP. The comparably weaker absorption signal can be improved by using a longer path-length flow cell. The FA-SEC profiles were congruent with the ones plotted by FSEC, suggesting FA-SEC could be a comparable and economical setup for detergent screening of membrane proteins. PMID:27332877

  10. Active Aeroelastic Wing Aerodynamic Model Development and Validation for a Modified F/A-18A Airplane

    NASA Technical Reports Server (NTRS)

    Cumming, Stephen B.; Diebler, Corey G.

    2005-01-01

    A new aerodynamic model has been developed and validated for a modified F/A-18A airplane used for the Active Aeroelastic Wing (AAW) research program. The goal of the program was to demonstrate the advantages of using the inherent flexibility of an aircraft to enhance its performance. The research airplane was an F/A-18A with wings modified to reduce stiffness and a new control system to increase control authority. There have been two flight phases. Data gathered from the first flight phase were used to create the new aerodynamic model. A maximum-likelihood output-error parameter estimation technique was used to obtain stability and control derivatives. The derivatives were incorporated into the National Aeronautics and Space Administration F-18 simulation, validated, and used to develop new AAW control laws. The second phase of flights was used to evaluate the handling qualities of the AAW airplane and the control law design process, and to further test the accuracy of the new model. The flight test envelope covered Mach numbers between 0.85 and 1.30 and dynamic pressures from 600 to 1250 pound-force per square foot. The results presented in this report demonstrate that a thorough parameter identification analysis can be used to improve upon models that were developed using other means. This report describes the parameter estimation technique used, details the validation techniques, discusses differences between previously existing F/A-18 models, and presents results from the second phase of research flights.

  11. Foodservice Occupations Cluster Guide.

    ERIC Educational Resources Information Center

    Oregon State Dept. of Education, Salem.

    Intended to assist vocational teachers in developing and implementing a cluster program in food service occupations, this guide contains sections on cluster organization and implementation and instructional emphasis areas. The cluster organization and implementation section covers goal-based planning and includes a proposed cluster curriculum, a…

  12. Cluster-impact fusion

    SciTech Connect

    Echenique, P.M.; Manson, J.R.; Ritchie, R.H. )

    1990-03-19

    We present a model for the cluster-impact-fusion experiments of Buehler, Friedlander, and Friedman, Calculated fusion rates as a function of bombarding energy for constant cluster size agree well with experiment. The dependence of the fusion rate on cluster size at fixed bombarding energy is explained qualitatively. The role of correlated, coherent collisions in enhanced energy loss by clusters is emphasized.

  13. Relationship between platelet phospholipid FA and mean platelet volume in healthy men.

    PubMed

    Li, Duo; Turner, Alan; Sinclair, Andrew J

    2002-09-01

    Increased mean platelet volume (MPV) has been suggested as an independent risk factor for acute myocardial infarction and the increased reactivity of large platelets. The aim of this study was to investigate the correlation between platelet phospholipid (PL) PUFA composition and MPV in 139 free-living healthy men ages 20-55 yr (vegans, n = 18; ovolacto vegetarians, n = 43; moderate meat-eaters, n = 60; and high meateaters, n = 18). Each subject completed a semiquantitative Food Frequency Questionnaire and gave a blood sample. Platelet PL FA composition and MPV were determined by standard methods. MPV was significantly greater in the vegans than in the ovolacto vegetarian, moderate, or high meat-eater groups (P < 0.01). Both vegan and ovolacto vegetarian groups had significantly higher platelet PL 18:2n-6 and 22:4n-6, and lower 20:5n-3 and 22:6n-3 compared with the moderate and high meat-eater groups. The vegans demonstrated a significant reduction in 20:4n-6 and 22:5n-3 compared with the ovolacto vegetarian, high meat-eater, and moderate meat-eater groups. Bivariate analysis results showed that MPV was significantly positively correlated with platelet PL 18:2n-6 (P = 0.048) and negatively correlated with 20:3n-6 (P = 0.02), 20:5n-3 (P = 0.005), and 22:5n-3 (P< 0.0001), respectively. In a multiple linear regression analysis, after controlling for potential confounding factors such as dietary group, age, exercise, body mass index, and dietary polyunsaturated and saturated fat, cholesterol, carbohydrate, and fiber intake, the MPV was still strongly negatively correlated with platelet PL 20:3n-6 (P = 0.003) and 22:5n-3 (P = 0.001). The present data suggest that 22:5n-3 and 20:3n-6 may play a role in the structural function of the platelet membrane. PMID:12458626

  14. Identification of a 12-gene Fusaric Acid Biosynthetic Gene Cluster in Fusarium Species Through Comparative and Functional Genomics.

    PubMed

    Brown, Daren W; Lee, Seung-Ho; Kim, Lee-Han; Ryu, Jae-Gee; Lee, Soohyung; Seo, Yunhee; Kim, Young Ho; Busman, Mark; Yun, Sung-Hwan; Proctor, Robert H; Lee, Theresa

    2015-03-01

    In fungi, genes involved in biosynthesis of a secondary metabolite (SM) are often located adjacent to one another in the genome and are coordinately regulated. These SM biosynthetic gene clusters typically encode enzymes, one or more transcription factors, and a transport protein. Fusaric acid is a polyketide-derived SM produced by multiple species of the fungal genus Fusarium. This SM is of concern because it is toxic to animals and, therefore, is considered a mycotoxin and may contribute to plant pathogenesis. Preliminary descriptions of the fusaric acid (FA) biosynthetic gene (FUB) cluster have been reported in two Fusarium species, the maize pathogen F. verticillioides and the rice pathogen F. fujikuroi. The cluster consisted of five genes and did not include a transcription factor or transporter gene. Here, analysis of the FUB region in F. verticillioides, F. fujikuroi, and F. oxysporum, a plant pathogen with multiple hosts, indicates the FUB cluster consists of at least 12 genes (FUB1 to FUB12). Deletion analysis confirmed that nine FUB genes, including two Zn(II)2Cys6 transcription factor genes, are required for production of wild-type levels of FA. Comparisons of FUB cluster homologs across multiple Fusarium isolates and species revealed insertion of non-FUB genes at one or two locations in some homologs. Although the ability to produce FA contributed to the phytotoxicity of F. oxysporum culture extracts, lack of production did not affect virulence of F. oxysporum on cactus or F. verticillioides on maize seedlings. These findings provide new insights into the genetic and biochemical processes required for FA production. PMID:25372119

  15. Cysteine Methylation Controls Radical Generation in the Cfr Radical AdoMet rRNA Methyltransferase

    PubMed Central

    Challand, Martin R.; Salvadori, Enrico; Driesener, Rebecca C.; Kay, Christopher W. M.; Roach, Peter L.; Spencer, James

    2013-01-01

    The ‘radical S-adenosyl-L-methionine (AdoMet)’ enzyme Cfr methylates adenosine 2503 of the 23S rRNA in the peptidyltransferase centre (P-site) of the bacterial ribosome. This modification protects host bacteria, notably methicillin-resistant Staphylococcus aureus (MRSA), from numerous antibiotics, including agents (e.g. linezolid, retapamulin) that were developed to treat such organisms. Cfr contains a single [4Fe-4S] cluster that binds two separate molecules of AdoMet during the reaction cycle. These are used sequentially to first methylate a cysteine residue, Cys338; and subsequently generate an oxidative radical intermediate that facilitates methyl transfer to the unreactive C8 (and/or C2) carbon centres of adenosine 2503. How the Cfr active site, with its single [4Fe-4S] cluster, catalyses these two distinct activities that each utilise AdoMet as a substrate remains to be established. Here, we use absorbance and electron paramagnetic resonance (EPR) spectroscopy to investigate the interactions of AdoMet with the [4Fe-4S] clusters of wild-type Cfr and a Cys338 Ala mutant, which is unable to accept a methyl group. Cfr binds AdoMet with high (∼ 10 µM) affinity notwithstanding the absence of the RNA cosubstrate. In wild-type Cfr, where Cys338 is methylated, AdoMet binding leads to rapid oxidation of the [4Fe-4S] cluster and production of 5'-deoxyadenosine (DOA). In contrast, while Cys338 Ala Cfr binds AdoMet with equivalent affinity, oxidation of the [4Fe-4S] cluster is not observed. Our results indicate that the presence of a methyl group on Cfr Cys338 is a key determinant of the activity of the enzyme towards AdoMet, thus enabling a single active site to support two distinct modes of AdoMet cleavage. PMID:23861844

  16. Survey on granularity clustering.

    PubMed

    Ding, Shifei; Du, Mingjing; Zhu, Hong

    2015-12-01

    With the rapid development of uncertain artificial intelligent and the arrival of big data era, conventional clustering analysis and granular computing fail to satisfy the requirements of intelligent information processing in this new case. There is the essential relationship between granular computing and clustering analysis, so some researchers try to combine granular computing with clustering analysis. In the idea of granularity, the researchers expand the researches in clustering analysis and look for the best clustering results with the help of the basic theories and methods of granular computing. Granularity clustering method which is proposed and studied has attracted more and more attention. This paper firstly summarizes the background of granularity clustering and the intrinsic connection between granular computing and clustering analysis, and then mainly reviews the research status and various methods of granularity clustering. Finally, we analyze existing problem and propose further research. PMID:26557926

  17. This modified F/A-18A with its distinctive red, white and blue paint scheme is the test aircraft for

    NASA Technical Reports Server (NTRS)

    2001-01-01

    This modified F/A-18A with its distinctive red, white and blue paint scheme is the test aircraft for the Active Aeroelastic Wing (AAW) project at NASA's Dryden Flight Research Center, Edwards, California.

  18. A modified F/A-18A sporting a distinctive red, white and blue paint scheme is the test aircraft for

    NASA Technical Reports Server (NTRS)

    2001-01-01

    A modified F/A-18A sporting a distinctive red, white and blue paint scheme is the test aircraft for the Active Aeroelastic Wing (AAW) project at NASA's Dryden Flight Research Center, Edwards, California.

  19. With a long flight data probe extending from its nose, this F/A-18A has been modified to conduct fli

    NASA Technical Reports Server (NTRS)

    2001-01-01

    With a long flight data probe extending from its nose, this F/A-18A has been modified to conduct flight research in the Active Aeroelastic Wing (AAW) project at NASA's Dryden Flight Research Center, Edwards, California.

  20. GNRs@SiO2-FA in combination with radiotherapy induces the apoptosis of HepG2 cells by modulating the expression of apoptosis-related proteins

    PubMed Central

    GAO, BIN; SHEN, LEI; HE, KE-WU; XIAO, WEI-HUA

    2015-01-01

    The aim of the present study was to examine the apoptosis of the hepatocellular carcinoma cell line, HepG2, induced by treatment with folic acid-conjugated silica-coated gold nanorods (GNRs@SiO2-FA) in combination with radiotherapy, and to determine the involvement of apoptosis-related proteins. An MTT colorimetric assay was used to assess the biocompatibility of GNRs@SiO2-FA. The distribution of GNRs@SiO2-FA into the cells was observed using transmission electron microscopy (TEM). HepG2 cells cultured in vitro were divided into the following 4 groups: i)the control group (untreated), ii) the GNRs@SiO2-FA group, iii) the radiotherapy group (iodine 125 seeds) and iv) the combination group (treated with GNRs@SiO2-FA and iodine 125 seeds) groups. The apoptosis of the HepG2 cells was detected by flow cytometry. The concentration range of <40 µg/ml GNRs@SiO2-FA was found to be safe for the biological activity of the HepG2 cells. GNRs@SiO2-FA entered the cytoplasm through endocytosis. The apoptotic rates of the HepG2 cells were higher in the GNRs@SiO2-FA and radiotherapy groups than in the control group (P<0.05). The apoptotic rate was also significantly higher in the combination group than the GNRs@SiO2-FA and radiotherapy groups (P<0.05). Taken together, these findings demonstrate that the combination of GNRs@SiO2-FA and radiotherapy more effectively induces the apoptosis of HepG2 cells. These apoptotic effects are achieved by increasing the protein expression of Bax and caspase-3, and inhibiting the protein expression of Bcl-2 and Ki-67. The combination of GNRs@SiO2-FA and radiotherapy may thus prove to be a new approach in the treatment of primary liver cancer. PMID:26648274

  1. GNRs@SiO₂-FA in combination with radiotherapy induces the apoptosis of HepG2 cells by modulating the expression of apoptosis-related proteins.

    PubMed

    Gao, Bin; Shen, Lei; He, Ke-Wu; Xiao, Wei-Hua

    2015-11-01

    The aim of the present study was to examine the apoptosis of the hepatocellular carcinoma cell line, HepG2, induced by treatment with folic acid-conjugated silica-coated gold nanorods (GNRs@SiO2-FA) in combination with radiotherapy, and to determine the involvement of apoptosis-related proteins. An MTT colorimetric assay was used to assess the biocompatibility of GNRs@SiO2-FA. The distribution of GNRs@SiO2-FA into the cells was observed using transmission electron microscopy (TEM). HepG2 cells cultured in vitro were divided into the following 4 groups: i)the control group (untreated), ii) the GNRs@SiO2-FA group, iii) the radiotherapy group (iodine 125 seeds) and iv) the combination group (treated with GNRs@SiO2-FA and iodine 125 seeds) groups. The apoptosis of the HepG2 cells was detected by flow cytometry. The concentration range of <40 µg/ml GNRs@SiO2-FA was found to be safe for the biological activity of the HepG2 cells. GNRs@SiO2-FA entered the cytoplasm through endocytosis. The apoptotic rates of the HepG2 cells were higher in the GNRs@SiO2-FA and radiotherapy groups than in the control group (P<0.05). The apoptotic rate was also significantly higher in the combination group than the GNRs@SiO2-FA and radiotherapy groups (P<0.05). Taken together, these findings demonstrate that the combination of GNRs@SiO2-FA and radiotherapy more effectively induces the apoptosis of HepG2 cells. These apoptotic effects are achieved by increasing the protein expression of Bax and caspase-3, and inhibiting the protein expression of Bcl-2 and Ki-67. The combination of GNRs@SiO2-FA and radiotherapy may thus prove to be a new approach in the treatment of primary liver cancer. PMID:26648274

  2. Glucomannan and glucomannan plus spirulina added to pork significantly block dietary cholesterol effects on lipoproteinemia, arylesterase activity, and CYP7A1 expression in Zucker fa/fa rats.

    PubMed

    González-Torres, Laura; Vázquez-Velasco, Miguel; Olivero-David, Raúl; Bastida, Sara; Benedí, Juana; González, Rafaela Raposo; González-Muñoz, Ma José; Sánchez-Muniz, Francisco J

    2015-12-01

    Zucker fa/fa rats easily develop dyslipidemia and obesity. Restructured pork (RP) is a suitable matrix for including functional ingredients. The effects of glucomannan- RP or glucomannan plus spirulina-enriched RP on plasma lipid/lipoprotein levels, cytochrome P450 7A1 (CYP7A1) expression, and arylesterase activity in growing fa/fa rats fed high-energy, high-fat cholesterol-enriched diets were tested. Groups of six rats each received diet containing 15% control-RP (C), 15% glucomannan-RP diet (G), 15% glucomannan + spirulina-RP diet (GS), and same diets enriched with 2.4% cholesterol and 0.49% cholic acid (cholesterol-enriched control (HC), cholesterol-enriched glucomannan (HG), and cholesterol-enriched glucomannan + spirulina (HGS) diets) over a 7-week period. C diet induced obesity, severe hyperglycemia, moderate hypercholesterolemia, and hypertriglyceridemia. Those facts were not significantly modified by G or GS diets. G diet increased CYP7A1 expression but decreased the total cholesterol/high density lipoproteins (HDL)-cholesterol ratio (p < 0.05) vs. C diet. GS vs. G diet increased (p < 0.05) CYP7A1 expression. HC vs. C diet reduced food intake, body weight gain, and plasma glucose (p < 0.01) but increased cholesterolemia (p < 0.01), lipidemia (plasma cholesterol plus triglycerides) (p < 0.001), cholesterol/triglyceride ratio in very low density lipoproteins (VLDL), and HDL (p < 0.05), cholesterol transported by VLDL and intermediate density lipoproteins (IDL) + low density lipoproteins (LDL), total cholesterol/HDL-cholesterol ratio and CYP7A1 expression (at least p < 0.05). HG and HGS diets vs. HC noticeably reduced lipidemia (p < 0.001), normalized VLDL and IDL + LDL lipid composition, and increased CYP7A1 expression (p < 0.01) but did not modify the cholesterol/HDL-cholesterol ratio. HGS vs. HG decreased triglyceridemia, the triglyceride-glucose (TyG) index and increased arylesterase/HDL-cholesterol activity (p < 0

  3. Cluster Morphology Analysis

    PubMed Central

    Jacquez, Geoffrey M.

    2009-01-01

    Most disease clustering methods assume specific shapes and do not evaluate statistical power using the applicable geography, at-risk population, and covariates. Cluster Morphology Analysis (CMA) conducts power analyses of alternative techniques assuming clusters of different relative risks and shapes. Results are ranked by statistical power and false positives, under the rationale that surveillance should (1) find true clusters while (2) avoiding false clusters. CMA then synthesizes results of the most powerful methods. CMA was evaluated in simulation studies and applied to pancreatic cancer mortality in Michigan, and finds clusters of flexible shape while routinely evaluating statistical power. PMID:20234799

  4. Determination of the stability and control derivatives of the F/A-18 HARV from flight data using the maximum likelihood method

    NASA Technical Reports Server (NTRS)

    Napolitano, Marcello R.; Spagnuolo, Joelle M.

    1992-01-01

    The research being conducted pertains to the determination of the stability and control derivatives of the F/A-18 High Alpha Research Vehicle (HARV) from flight data using the Maximum Likelihood Method. The document outlines the approach used in the parameter estimation (PID) process and briefly describes the mathematical modeling of the F/A-18 HARV and the maneuvers designed to generate a sufficient data base for the PID research.

  5. "Near-bottom sonar mapping along the Eastern Lau Spreading Center and Valu Fa Ridge"

    NASA Astrophysics Data System (ADS)

    Sleeper, J. D.; Martinez, F.; Edwards, M.; Dunn, R.; Baker, E. T.

    2009-12-01

    We present results from high resolution sidescan sonar mapping at the Ridge 2000 Lau Integrated Studies Site, a back-arc spreading center in the Southwest Pacific. The survey covered segments of the Eastern Lau Spreading Center (ELSC) and Valu Fa Ridge (VFR), which exhibit strong contrasts in geologic and geophysical characteristics, including variations in ridge morphology, volcanic and tectonic structures, lava composition and texture, magma production, and spreading rate. The bulk of the data were collected in 2008 aboard R/V Kilo Moana (KM0804), using the deep-towed IMI120 (DSL120A) sonar, with an array of miniature autonomous plume recorder's (MAPR's) tethered to the tow cable to detect hydrothermal plume signatures in the water column. The ELSC survey covered an area ~10 km wide by 100 km long centered on the spreading axis, with tracks spaced ~1 km apart, but as close as ~500 m in the vicinity of the ABE hydrothermal vent field, and also included the Tow Cam and Kilo Moana vent fields. The VFR survey covered a ~5 km wide by 100 km long area with ~700 m-spaced tracks, including the Vai Lili, Mariner, and Tui Malila vent sites. The IMI120 has a resolution of ~1-2 meters, giving the first detailed view of the on- and off-axis structures in the two focus regions. Navigational corrections to the sidescan data were made through feature-matching between the deep-towed data and a lower resolution background ship bathymetry grid, using software developed by the Hawaiian Mapping and Research Group. Background bathymetry and lower resolution (~100 m) sidescan data were compiled from previous cruises on R/V Kilo Moana, and an early-2009 seismic survey on R/V Marcus G. Langseth (L-SCAN). Hydrothermal plume data collected with the MAPR's are coregistered with the deep-towed sidescan data to illuminate correlations between seafloor structures and plume signatures. The axis of the VFR in the south forms a peaked ridge, inflated by abundant melt supply, and draped with

  6. Thermodynamics of the Electron Acceptors in Heliobacterium modesticaldum: An Exemplar of an Early Homodimeric Type I Photosynthetic Reaction Center.

    PubMed

    Ferlez, Bryan; Cowgill, John; Dong, Weibing; Gisriel, Christopher; Lin, Su; Flores, Marco; Walters, Karim; Cetnar, Daniel; Redding, Kevin E; Golbeck, John H

    2016-04-26

    The homodimeric type I reaction center in heliobacteria is arguably the simplest known pigment-protein complex capable of conducting (bacterio)chlorophyll-based conversion of light into chemical energy. Despite its structural simplicity, the thermodynamics of the electron transfer cofactors on the acceptor side have not been fully investigated. In this work, we measured the midpoint potential of the terminal [4Fe-4S](2+/1+) cluster (FX) in reaction centers from Heliobacterium modesticaldum. The FX cluster was titrated chemically and monitored by (i) the decrease in the level of stable P800 photobleaching by optical spectroscopy, (ii) the loss of the light-induced g ≈ 2 radical from P800(+•) following a single-turnover flash, (iii) the increase in the low-field resonance at 140 mT attributed to the S = (3)/2 ground spin state of FX(-), and (iv) the loss of the spin-correlated P800(+) FX(-) radical pair following a single-turnover flash. These four techniques led to similar estimations of the midpoint potential for FX of -502 ± 3 mV (n = 0.99), -496 ± 2 mV (n = 0.99), -517 ± 10 mV (n = 0.65), and -501 ± 4 mV (n = 0.84), respectively, with a consensus value of -504 ± 10 mV (converging to n = 1). Under conditions in which FX is reduced, the long-lived (∼15 ms) P800(+) FX(-) state is replaced by a rapidly recombining (∼15 ns) P800(+)A0(-) state, as shown by ultrafast optical experiments. There was no evidence of the presence of a P800(+) A1(-) spin-correlated radical pair by electron paramagnetic resonance (EPR) under these conditions. The midpoint potentials of the two [4Fe-4S](2+/1+) clusters in the low-molecular mass ferredoxins were found to be -480 ± 11 mV/-524 ± 13 mV for PshBI, -453 ± 6 mV/-527 ± 6 mV for PshBII, and -452 ± 5 mV/-533 ± 8 mV for HM1_2505 as determined by EPR spectroscopy. FX is therefore suitably poised to reduce one [4Fe-4S](2+/1+) cluster in these mobile electron carriers. Using the measured midpoint potential of FX and a

  7. Fasted-state simulated intestinal fluid "FaSSIF-C", a cholesterol containing intestinal model medium for in vitro drug delivery development.

    PubMed

    Khoshakhlagh, Pooneh; Johnson, Raphael; Langguth, Peter; Nawroth, Thomas; Schmueser, Lars; Hellmann, Nadja; Decker, Heinz; Szekely, Noemi Kinga

    2015-07-01

    A set of biorelevant media "fasted-state simulated intestinal fluid with cholesterol (FaSSIF-C)" for the in vitro study of intestinal drug dissolution in the duodenum was developed. These contain cholesterol at the same levels as in human bile: the cholesterol content of FaSSIF-7C is equivalent to healthy female, FaSSIF-10C to healthy male persons, and FaSSIF-13C to several disease cases that lead to gallstones. The fluids were studied in three aspects: biocompatibility, intestinal nanostructure, and solubilizing power of hydrophobic drugs of the BCS class II. The biocompatibility study showed no toxic effects in a Caco-2 cell system. The drug-solubilizing capacity toward Fenofibrate, Danazol, Griseofulvin, and Carbamazepine was assessed as example. It varied with the cholesterol content widely from a fourfold improvement to a twofold reduction. The nanostructure study by dynamic light scattering and small-angle neutron scattering indicated vesicles as the main component of FaSSIF-C in equilibrium (>1 h), but at high cholesterol content, larger particles were observed as a minor contribution. The neutron experiments indicated the presence of complex micelle-vesicle mixtures, even after 1 h development of fed-state bile model to FaSSIF. The results indicate that cholesterol affects some drugs in solubilization and particle size in intestinal model fluids. PMID:25964103

  8. Universal Cluster Deposition System

    NASA Astrophysics Data System (ADS)

    Qiang, You; Sun, Zhiguang; Sellmyer, David J.

    2001-03-01

    We have developed a universal cluster deposition system (UCDS), which combines a new kind of sputtering-gas-aggregation (SGA) cluster beam source with two atom beams from magnetron sputtering. A highly intense, very stable beam of nanoclusters (like Co, Fe, Ni, Si, CoSm or CoPt) are produced. A quadrupole and/or a new high transmission infinite range mass selector have been designed for the cluster beam. The size distribution (Δd/d) is between 0.05+/-0.10, measured in situ by TOF. A range of mean cluster size is 2 to 10 nm. Usually the deposition rate is about 5 deg/s. The cluster concentration in the film is adjusted through the ratio of cluster and atomic beam deposition rates, as measured in situ with a rotatable quartz microbalance. The UCDS can be used to prepare coated clusters. After exiting from the cluster source, the clusters can be coated first with an atomic or molecular species in an evaporation chamber, and deposited alone or co-deposited with another material. This system is used to deposit simultaneously or alternately mesoscopic thin films or multilayers, and offers the possibility to control independently the incident cluster size and concentration, and thereby the interaction between clusters and cluster-matrix material which is of interest for fundamental research and industry applications. Magnetic properties of Co cluster-assembled materials will be discussed. * Research supported by NSF, DARPA through ARO, and CMRA

  9. Metabolic Interaction between Anthocyanin and Lignin Biosynthesis Is Associated with Peroxidase FaPRX27 in Strawberry Fruit1[W

    PubMed Central

    Ring, Ludwig; Yeh, Su-Ying; Hücherig, Stephanie; Hoffmann, Thomas; Blanco-Portales, Rosario; Fouche, Mathieu; Villatoro, Carmen; Denoyes, Béatrice; Monfort, Amparo; Caballero, José Luis; Muñoz-Blanco, Juan; Gershenson, Jonathan; Schwab, Wilfried

    2013-01-01

    Plant phenolics have drawn increasing attention due to their potential nutritional benefits. Although the basic reactions of the phenolics biosynthetic pathways in plants have been intensively analyzed, the regulation of their accumulation and flux through the pathway is not that well established. The aim of this study was to use a strawberry (Fragaria × ananassa) microarray to investigate gene expression patterns associated with the accumulation of phenylpropanoids, flavonoids, and anthocyanins in strawberry fruit. An examination of the transcriptome, coupled with metabolite profiling data from different commercial varieties, was undertaken to identify genes whose expression correlated with altered phenolics composition. Seventeen comparative microarray analyses revealed 15 genes that were differentially (more than 200-fold) expressed in phenolics-rich versus phenolics-poor varieties. The results were validated by heterologous expression of the peroxidase FaPRX27 gene, which showed the highest altered expression level (more than 900-fold). The encoded protein was functionally characterized and is assumed to be involved in lignin formation during strawberry fruit ripening. Quantitative trait locus analysis indicated that the genomic region of FaPRX27 is associated with the fruit color trait. Down-regulation of the CHALCONE SYNTHASE gene and concomitant induction of FaPRX27 expression diverted the flux from anthocyanins to lignin. The results highlight the competition of the different phenolics pathways for their common precursors. The list of the 15 candidates provides new genes that are likely to impact polyphenol accumulation in strawberry fruit and could be used to develop molecular markers to select phenolics-rich germplasm. PMID:23835409

  10. Development and verification of real-time controllers for the F/A-18 vertical fin buffet load alleviation

    NASA Astrophysics Data System (ADS)

    Chen, Yong; Viresh, Wickramasinghe; Zimcik, David

    2006-03-01

    Twin-tail fighter aircraft such as the F/A-18 may experience intense buffet loads at high angles of attack flight conditions and the broadband buffet loads primarily excite the first bending and torsional modes of the vertical fin that results in severe vibration and dynamic stresses on the vertical fin structures. To reduce the premature fatigue failure of the structure and to increase mission availability, a novel hybrid actuation system was developed to actively alleviate the buffet response of a full-scale F/A-18 vertical fin. A hydraulic rudder actuator was used to control the bending mode of the fin by engaging the rudder inertial force. Multiple Macro Fiber Composites actuators were surface mounted to provide induced strain actuation authority to control the torsional mode. Experimental system identification approach was selected to obtain a state-space model of the system using open-loop test data. An LQG controller was developed to minimize the dynamic response of the vertical fin at critical locations. Extensive simulations were conducted to evaluate the control authority of the actuators and the performance of the controller under various buffet load cases and levels. Closed-loop tests were performed on a full-scale F/A-18 empennage and the results validated the effectiveness of the real-time controller as well as the development methodology. In addition, the ground vibration test demonstrated that the hybrid actuation system is a feasible solution to alleviate the vertical tail buffet loads in high performance fighter aircraft.

  11. In Situ Generation of Active Molybdenum Octahedral Clusters for Photocatalytic Hydrogen Production from Water.

    PubMed

    Feliz, Marta; Puche, Marta; Atienzar, Pedro; Concepción, Patricia; Cordier, Stéphane; Molard, Yann

    2016-08-01

    The photocatalytic hydrogen evolution reaction (HER) from water under homogeneous and heterogeneous conditions is explored for the {Mo6 Br(i) 8 }(4+) cluster core based unit starting from (TBA)2 [Mo6 Br(i) 8 F(a) 6 ] (TBA=tetra-n-butylammonium; "i" and "a" refer to the face-capping inner and terminal apical ligand, respectively). The catalytic activity of {Mo6 Br(i) 8 }(4+) is enhanced by the in situ generation of [Mo6 Br(i) 8 F(a) 5 (OH)(a) ](2-) , [Mo6 Br(i) 8 F(a) 3 (OH)(a) 3 ](2-) , and [Mo6 Br(i) 8 (OH)(a) 6 ](2-) , which are identified by ESIMS, luminescence, and NMR techniques. Full substitution of F(-) by OH(-) leads to the formation of (H3 O)2 [Mo6 Br(i) 8 (OH)(a) 6 ]⋅10 H2 O; its structure was determined by single-crystal XRD. The immobilization of the active {Mo6 Br(i) 8 }(4+) onto graphene oxide (GO) surfaces enhances its stability under catalytic conditions. The catalytic activity of the resulting (TBA)2 Mo6 Br(i) 8 @GO material is improved with respect to GO, but is reduced compared to the activity under homogeneous conditions because of changes in the GO semiconducting properties as well as lower activity and/or accessibility of the anchored cluster. PMID:27314221

  12. New chemical alternative for ozone-depleting substances: HFC-236fa. Final report, December 1996-April 1997

    SciTech Connect

    Smith, N.D.; Brna, T.G.; Gage, C.L.; Hendriks, R.V.

    1997-07-01

    The report gives results of a preliminary evaluation of a new hydrofluoro-carbon (HFC)--HFC-236fa or 1,1,1,3,3,3-hexafluoropropane--asa possible alternative for chlorofluorocarbon (CFC)-114 (1,2-dichloro-1, 1,2,2-tetrafluoroethane) refrigerant for chillers and asa possible fire suppressant replacement for halon-1301 (bromotrifluoromethane). Evaluation tests included an examination of flammability, stability, atmospheric lifetime, thermophysical properties, lubricant miscibility and solubility, materials compatibility, inhalation toxicity, refrigerant performance, heat transfer characteristics, and flame suppression.

  13. Alternatives to CFC-114 in high-temperature heat pumps: Compressor performance with HFC-236ea and HFC-236fa

    SciTech Connect

    Kazachki, G S; Gage, C L; Hendriks, R V

    1996-01-01

    The paper gives results of comprehensive calorimeter tests on a semihermetic compressor with chlorofluorocarbon (CFC)-114, hydrofluorocarbon (HFC)-236ea, and HFC-236fa over a wide range of temperature test conditions: evaporating temperatures from 0 to 35 C and condensing temperatures from 40 to 110 C. Parameters assessed as criteria for performance evaluation and for reliable performance include: cooling capacity; electric power input, current, and voltage; coefficients of performance; compressor volumetric and isentropic efficiencies; and discharge and oil temperatures. Polyolester oil was used as lubricant in the compressor.

  14. Wind tunnel measurements on a full-scale F/A-18 with forebody slot blowing or forebody strakes

    NASA Technical Reports Server (NTRS)

    Lanser, Wendy R.; Murri, Daniel G.

    1993-01-01

    Results are presented of tests, conducted on a full-scale F/A-18 in the 120-Foot Wind Tunnel at NASA Ames Research Center, to measure the effectiveness of a 16-in.-long tangentially blown slot and of deployable strakes (measuring 4 ft in length) positioned on the aircraft's forebody. Fixed strakes with deflections of 30, 60, or 90 deg were tested to simulate the deployment of conformal actuated forebody strakes. It is shown that both the tangentially blown slot and the deployable strakes are effective in generating large yawing momemts at high angles of attack, without inducing significant coupling in the other axes.

  15. EINSTEIN Cluster Alignments Revisited

    NASA Astrophysics Data System (ADS)

    Chambers, S. W.; Melott, A. L.; Miller, C. J.

    2000-12-01

    We have examined whether the major axes of rich galaxy clusters tend to point (in projection) toward their nearest neighboring cluster. We used the data of Ulmer, McMillan and Kowalski, who used x-ray morphology to define position angles. Our cluster samples, with well measured redshifts and updated positions, were taken from the MX Northern Abell Cluster Survey. The usual Kolmogorov-Smirnov test shows no significant alignment signal for nonrandom angles for all separations less than 100 Mpc/h. Refining the null hypothesis, however, with the Wilcoxon rank-sum test, reveals a high confidence signal for alignment. This confidence is highest when we restrict our sample to small nearest neighbor separations. We conclude that we have identified a more powerful tool for testing cluster-cluster alignments. Moreover, there is a strong signal in the data for alignment, consistent with a picture of hierarchical cluster formation in which matter falls into clusters along large scale filamentary structures.

  16. Matlab Cluster Ensemble Toolbox

    SciTech Connect

    Sapio, Vincent De; Kegelmeyer, Philip

    2009-04-27

    This is a Matlab toolbox for investigating the application of cluster ensembles to data classification, with the objective of improving the accuracy and/or speed of clustering. The toolbox divides the cluster ensemble problem into four areas, providing functionality for each. These include, (1) synthetic data generation, (2) clustering to generate individual data partitions and similarity matrices, (3) consensus function generation and final clustering to generate ensemble data partitioning, and (4) implementation of accuracy metrics. With regard to data generation, Gaussian data of arbitrary dimension can be generated. The kcenters algorithm can then be used to generate individual data partitions by either, (a) subsampling the data and clustering each subsample, or by (b) randomly initializing the algorithm and generating a clustering for each initialization. In either case an overall similarity matrix can be computed using a consensus function operating on the individual similarity matrices. A final clustering can be performed and performance metrics are provided for evaluation purposes.

  17. [Pathophysiology of cluster headache].

    PubMed

    Donnet, Anne

    2015-11-01

    The aetiology of cluster headache is partially unknown. Three areas are involved in the pathogenesis of cluster headache: the trigeminal nociceptive pathways, the autonomic system and the hypothalamus. The cluster headache attack involves activation of the trigeminal autonomic reflex. A dysfunction located in posterior hypothalamic gray matter is probably pivotal in the process. There is a probable association between smoke exposure, a possible genetic predisposition and the development of cluster headache. PMID:26470883

  18. Design of an effective bifunctional catalyst organotriphosphonic acid-functionalized ferric alginate (ATMP-FA) and optimization by Box-Behnken model for biodiesel esterification synthesis of oleic acid over ATMP-FA.

    PubMed

    Liu, Wei; Yin, Ping; Liu, Xiguang; Qu, Rongjun

    2014-12-01

    Biodiesel production has become an intense research area because of rapidly depleting energy reserves and increasing petroleum prices together with environmental concerns. This paper focused on the optimization of the catalytic performance in the esterification reaction of oleic acid for biodiesel production over the bifunctional catalyst organotriphosphonic acid-functionalized ferric alginate ATMP-FA. The reaction parameters including catalyst amount, ethanol to oleic acid molar ratio and reaction temperature have been optimized by response surface methodology (RSM) using the Box-Behnken model. It was found that the reaction temperature was the most significant factor, and the best conversion ratio of oleic acid could reach 93.17% under the reaction conditions with 9.53% of catalyst amount and 8.62:1 of ethanol to oleic acid molar ratio at 91.0 °C. The research results show that two catalytic species could work cooperatively to promote the esterification reaction, and the bifunctional ATMP-FA is a potential catalyst for biodiesel production. PMID:25310862

  19. Electrospray Quadrupole Travelling Wave Ion Mobility Time-of-Flight Mass Spectrometry for the Detection of Plasma Metabolome Changes Caused by Xanthohumol in Obese Zucker (fa/fa) Rats

    PubMed Central

    Wickramasekara, Samanthi I.; Zandkarimi, Fereshteh; Morré, Jeff; Kirkwood, Jay; Legette, LeeCole; Jiang, Yuan; Gombart, Adrian F.; Stevens, Jan F.; Maier, Claudia S.

    2013-01-01

    This study reports on the use of traveling wave ion mobility quadrupole time-of-flight (ToF) mass spectrometry for plasma metabolomics. Plasma metabolite profiles of obese Zucker fa/fa rats were obtained after the administration of different oral doses of Xanthohumol; a hop-derived dietary supplement. Liquid chromatography coupled data independent tandem mass spectrometry (LC-MSE) and LC-ion mobility spectrometry (IMS)-MSE acquisitions were conducted in both positive and negative modes using a Synapt G2 High Definition Mass Spectrometry (HDMS) instrument. This method provides identification of metabolite classes in rat plasma using parallel alternating low energy and high energy collision spectral acquisition modes. Data sets were analyzed using pattern recognition methods. Statistically significant (p < 0.05 and fold change (FC) threshold > 1.5) features were selected to identify the up-/down-regulated metabolite classes. Ion mobility data visualized using drift scope software provided a graphical read-out of differences in metabolite classes. PMID:24958146

  20. Acute effects of food, 2-deoxy-D-glucose and noradrenaline on metabolic rate and brown adipose tissue in normal and atropinised lean and obese (fa/fa) Zucker rats.

    PubMed

    Rothwell, N J; Saville, M E; Stock, M J

    1981-12-01

    1. Intragastric feeding (40 kJ) produced a 17% rise in metabolic rate in lean Zucker rats but only an 8% increase in obese (fa/fa) rats, and both of these responses were significantly reduced by beta-adrenergic blockade with propranolol (10 mg/kg, s.c.). 2. Parasympathetic blockade with atropine (0.5 mg/kg, s.c.) caused a doubling of the response to food in lean rats and a threefold increase in the obese mutants, such that all atropinised animals showed the same increase in metabolic rate after food. 3. Feeding also caused a significant rise in interscapular brown adipose tissue temperature, which was greatest in the lean animals and was enhanced by atropine in both groups. 4. Injection of noradrenaline (250 micrograms/kg, s.c.) caused a similar (40%) rise in metabolic rate in lean and obese animals but this response was unaffected by atropine. 5. 2-Deoxy-D-glucose injection (360 mg/kg, s.c.) depressed oxygen consumption by 25 and 8% in lean and obese rats respectively and this effect was totally abolished by atropine. 6. These results suggest that the rise in metabolic rate after a meal is partly due to sympathetic activation of brown adipose tissue. The reduced thermic response in obese Zucker rats is not due to insensitivity to noradrenaline, but may be partly due to parasympathetic inhibition of thermogenesis and partly to insensitivity to glucose availability. PMID:7322844

  1. Participation of the iron-sulphur cluster and of the covalently bound coenzyme of trimethylamine dehydrogenase in catalysis.

    PubMed Central

    Steenkamp, D J; Singer, T P

    1978-01-01

    Bacterial trimethylamine dehydrogenase contains a novel type of covalently bound flavin mononucleotide and a tetrameric iron-sulphur centre. The dehydrogenase takes up 1.5mol of dithionite/mol of enzyme and is thereby converted into the flavin quinol-reduced (4Fe-4S) form, with the expected bleaching of the visible absorption band of the flavin and the emergence of signals of typical reduced ferredoxin in the electronparamagnetic-resonance spectrum. On reduction with a slight excess of substrate, however, unusual absorption and electron-paramagnetic-resonance spectra appear quite rapidly. The latter is attributed to extensive interaction between the reduced (4Fe-4S) centre and the flavin semiquinone. The species of enzyme arising during the catalytic cycle were studied by a combination of rapid-freeze e.p.r. and stopped-flow spectophotometry. The initial reduction of the flavin to the quinol form is far too rapid to be rate-limiting in catalysis, as is the reoxidation of the substrate-reduced enzyme by phenazine methosulphate. Formation of the spin-spin-interacting species from the dihydroflavin is considerably slower, however, and it may be the rate-limiting step in the catalytic cycle, since its rate of formation agrees reasonably well with the catalytic-centre activity determined in steady-state kinetic assays. In addition to the interacting form, a second form of the enzyme was noted during reduction by trimethylamine, differing in absorption spectrum, the structure of which remains to be determined. PMID:204297

  2. Participation of the iron-sulphur cluster and of the covalently bound coenzyme of trimethylamine dehydrogenase in catalysis.

    PubMed

    Steenkamp, D J; Singer, T P

    1978-02-01

    Bacterial trimethylamine dehydrogenase contains a novel type of covalently bound flavin mononucleotide and a tetrameric iron-sulphur centre. The dehydrogenase takes up 1.5mol of dithionite/mol of enzyme and is thereby converted into the flavin quinol-reduced (4Fe-4S) form, with the expected bleaching of the visible absorption band of the flavin and the emergence of signals of typical reduced ferredoxin in the electronparamagnetic-resonance spectrum. On reduction with a slight excess of substrate, however, unusual absorption and electron-paramagnetic-resonance spectra appear quite rapidly. The latter is attributed to extensive interaction between the reduced (4Fe-4S) centre and the flavin semiquinone. The species of enzyme arising during the catalytic cycle were studied by a combination of rapid-freeze e.p.r. and stopped-flow spectophotometry. The initial reduction of the flavin to the quinol form is far too rapid to be rate-limiting in catalysis, as is the reoxidation of the substrate-reduced enzyme by phenazine methosulphate. Formation of the spin-spin-interacting species from the dihydroflavin is considerably slower, however, and it may be the rate-limiting step in the catalytic cycle, since its rate of formation agrees reasonably well with the catalytic-centre activity determined in steady-state kinetic assays. In addition to the interacting form, a second form of the enzyme was noted during reduction by trimethylamine, differing in absorption spectrum, the structure of which remains to be determined. PMID:204297

  3. CLUSTERING OF RARE EVENTS

    EPA Science Inventory

    The clustering of cases of a rare disease is considered. The number of events observed for each unit is assumed to have a Poisson distribution, the mean of which depends upon the population size and the cluster membership of that unit. Here a cluster consists of those units that ...

  4. Rigorous procedure for mapping thermal infrared images on three-dimensional models of building façades

    NASA Astrophysics Data System (ADS)

    Previtali, Mattia; Barazzetti, Luigi; Redaelli, Veronica; Scaioni, Marco; Rosina, Elisabetta

    2013-01-01

    A rigorous methodology for mapping thermal and RGB images on three-dimensional (3-D) models of building façades is presented. The developed method differs from most existing approaches because it relies on the use of thermal images coupled with 3-D models derived from terrestrial laser scanning surveying. The primary issue for an accurate texturing is the coregistration of the geometric model of the façade and the thermal images in the same reference system. This task is done by using a procedure standing out from other approaches adopted in current practice, which are mainly based on the independent registration of each image on the basis of homography or space resection techniques. A rigorous photogrammetric orientation of both thermal and RGB images is computed together in a combined bundle adjustment. This solution allows one to have a better control of the quality of the results, especially to reduce errors and artifacts in areas where more images are mosaicked onto the 3-D model. Several products can be obtained: 3-D triangulated textured models or raster products like orthophotos, having the temperature as radiometric value. The proposed approach is tested on different buildings of Politecnico di Milano University. Applications demonstrated the performance of the procedure and its technical applicability in routine thermal surveys.

  5. Genetic modifiers of Lepr{sup fa} associated with variability in insulin production and susceptibility to NIDDM

    SciTech Connect

    Chung, W.K.; Zheng, M.; Chua, M.

    1997-05-01

    In an attempt to identify the genetic basis for susceptibility to non-insulin-dependent diabetes mellitus within the context of obesity, we generated 401 genetically obese Lepr{sup fa}/Lepr{sup fa} F2 WKY13M intercross rats that demonstrated wide variation in multiple phenotypic measures related to diabetes, including plasma glucose concentration, percentage of glycosylated hemoglobin, plasma insulin concentration, and pancreatic islet morphology. Using selective genotyping genome scanning approaches, we have identified three quantitative trait loci (QTLs) on Chr. 1 (LOD 7.1 for pancreatic morpholology), Chr. 12 (LOD 5.1 for body mass index and LOD 3.4 for plasma glucose concentration), and Chr. 16 (P < 0.001 for genotype effect on plasma glucose concentration). The obese F2 progeny demonstrated sexual dimorphism for these traits, with increased diabetes susceptibility in the males appearing at approximately 6 weeks of age, as sexual maturation occurred. For each of the QTLs, the linked phenotypes demonstrated sexual dimorphism (more severe affection in males). The QTL on Chr. 1 maps to a region vicinal to that previously linked to adiposity in studies of diabetes susceptibility in the nonobese Goto-Kakizaki rat, which is genetically closely related to the Wistar counterstrain we employed. Several candidate genes, including tubby (tub), multigenic obesity 1 (Mob1), adult obesity and diabetes (Ad), and insulin-like growth factor-2 (Igf2), map to murine regions homologous to the QTL region identified on rat Chr. 1. 60 refs., 5 figs., 4 tabs.

  6. The mechanisms underlying overgeneral autobiographical memory: an evaluative review of evidence for the CaR-FA-X model.

    PubMed

    Sumner, Jennifer A

    2012-02-01

    Overgeneral autobiographical memory (OGM) has been found to be an important cognitive phenomenon with respect to depression and trauma-related psychopathology (e.g., posttraumatic stress disorder), and researchers have been interested in better understanding the factors that contribute to this proposed vulnerability factor. The most prominent model of mechanisms underlying OGM to date is Williams et al.'s (2007) CaR-FA-X model. This model proposes that three processes influence OGM: capture and rumination, functional avoidance, and impaired executive control. The author reviews the current state of support for the CaR-FA-X model by evaluating 38 studies that have examined OGM and one or more mechanisms of the model. Collectively, these studies reveal robust support for associations between OGM and both rumination and impaired executive control. OGM also appears to be a cognitive avoidance strategy, and there is evidence that avoiding the retrieval of specific memories reduces distress after an aversive event, at least in the short term. Important issues that have been left unresolved are highlighted, including the nature of the capture phenomenon, the role of trauma in functional avoidance, and the developmental nature of functional avoidance. Recommendations for future research that will enhance understanding of the factors that contribute to OGM are suggested. PMID:22142837

  7. The mechanisms underlying overgeneral autobiographical memory: An evaluative review of evidence for the CaR-FA-X model

    PubMed Central

    Sumner, Jennifer A.

    2011-01-01

    Overgeneral autobiographical memory (OGM) has been found to be an important cognitive phenomenon with respect to depression and trauma-related psychopathology (e.g., posttraumatic stress disorder), and researchers have been interested in better understanding the factors that contribute to this proposed vulnerability factor. The most prominent model of mechanisms underlying OGM to date is Williams et al.’s (2007) CaR-FA-X model. This model proposes that three processes influence OGM: capture and rumination, functional avoidance, and impaired executive control. The author reviews the current state of support for the CaR-FA-X model by evaluating 38 studies that have examined OGM and one or more mechanisms of the model. Collectively, these studies reveal robust support for associations between OGM and both rumination and impaired executive control. OGM also appears to be a cognitive avoidance strategy, and there is evidence that avoiding the retrieval of specific memories reduces distress after an aversive event, at least in the short term. Important issues that have been left unresolved are highlighted, including the nature of the capture phenomenon, the role of trauma in functional avoidance, and the developmental nature of functional avoidance. Recommendations for future research that will enhance understanding of the factors that contribute to OGM are suggested. PMID:22142837

  8. Microbial deterioration of artistic tiles from the façade of the Grande Albergo Ausonia & Hungaria (Venice, Italy).

    PubMed

    Giacomucci, Lucia; Bertoncello, Renzo; Salvadori, Ornella; Martini, Ilaria; Favaro, Monica; Villa, Federica; Sorlini, Claudia; Cappitelli, Francesca

    2011-08-01

    The Grande Albergo Ausonia & Hungaria (Venice Lido, Italy) has an Art Nouveau polychrome ceramic coating on its façade, which was restored in 2007. Soon after the conservation treatment, many tiles of the façade decoration showed coloured alterations putatively attributed to the presence of microbial communities. To confirm the presence of the biological deposit and the stratigraphy of the Hungaria tiles, stereomicroscope, optical and environmental scanning electron microscope observations were made. The characterisation of the microbial community was performed using a PCR-DGGE approach. This study reported the first use of a culture-independent approach to identify the total community present in biodeteriorated artistic tiles. The case study examined here reveals that the coloured alterations on the tiles were mainly due to the presence of cryptoendolithic cyanobacteria. In addition, we proved that the microflora present on the tiles was generally greatly influenced by the environment of the Hungaria hotel. We found several microorganisms related to the alkaline environment, which is in the range of the tile pH, and related to the aquatic environment, the presence of the acrylic resin Paraloid B72® used during the 2007 treatment and the pollutants of the Venice lagoon. PMID:21286701

  9. Transonic Free-To-Roll Analysis of the F/A-18E and F-35 Configurations

    NASA Technical Reports Server (NTRS)

    Owens, D. Bruce; McConnell, Jeffrey K.; Brandon, Jay M.; Hall, Robert M.

    2004-01-01

    The free-to-roll technique is used as a tool for predicting areas of uncommanded lateral motions. Recently, the NASA/Navy/Air Force Abrupt Wing Stall Program extended the use of this technique to the transonic speed regime. Using this technique, this paper evaluates various wing configurations on the pre-production F/A-18E aircraft and the Joint Strike Fighter (F-35) aircraft. The configurations investigated include leading and trailing edge flap deflections, fences, leading edge flap gap seals, and vortex generators. These tests were conducted in the NASA Langley 16-Foot Transonic Tunnel. The analysis used a modification of a figure-of-merit developed during the Abrupt Wing Stall Program to discern configuration effects. The results showed how the figure-of-merit can be used to schedule wing flap deflections to avoid areas of uncommanded lateral motion. The analysis also used both static and dynamic wind tunnel data to provide insight into the uncommanded lateral behavior. The dynamic data was extracted from the time history data using parameter identification techniques. In general, modifications to the pre-production F/A-18E resulted in shifts in angle-of-attack where uncommanded lateral activity occurred. Sealing the gap between the inboard and outboard leading-edge flaps on the Navy version of the F-35 eliminated uncommanded lateral activity or delayed the activity to a higher angle-of-attack.

  10. C-axis electrical resistivity of PrO1-aFaBiS2 single crystals

    NASA Astrophysics Data System (ADS)

    Nagao, Masanori; Miura, Akira; Watauchi, Satoshi; Takano, Yoshihiko; Tanaka, Isao

    2015-08-01

    The high anisotropy in RO1-aFaBiS2 (R denotes a rare-earth element) superconductors demonstrates their potential use as intrinsic Josephson junctions, considering the weak coupling among BiS2-PrO(F)-BiS2 (superconducting-normal-superconducting) layers along the c-axis. We grew PrO1-aFaBiS2 single crystals using CsCl/KCl flux. The superconducting anisotropies of the grown single crystals were estimated to be approximately 40-50 from the effective mass model. The c-axis transport properties were characterized using single-crystal s-shaped intrinsic Josephson junctions with a focused ion beam. Along the c-axis, the crystals showed zero resistivity at 2.7 K and a critical current density of 1.33 × 103 A/cm2 at 2.0 K. The current-voltage curve along the c-axis displayed hysteresis. The c-axis transport measurements under a magnetic field parallel to the ab-plane revealed a “lock-in” state due to the Josephson vortex flow, indicating that BiS2 superconductors are promising candidates for intrinsic Josephson junctions.

  11. Flying Quality Analysis of a JAS 39 Gripen Ministick Controller in an F/A-18 Aircraft

    NASA Technical Reports Server (NTRS)

    Carter, John F.; Stoliker, P. C.

    2000-01-01

    NASA Dryden conducted a handling qualities experiment using a small displacement centerstick controller that Saab-Scania developed for the JAS 39 Gripen aircraft. The centerstick, or ministick, was mounted in the rear cockpit of an F/A-18 aircraft. Production support flight control computers (PSFCC) provided a pilot-selectable research control system. The objectives for this experiment included determining whether the mechanical characteristics of the centerstick controller had any significant effect on the handling qualities of the F/A-18, and determining the usefulness of the PSFCCs for this kind of experiment. Five pilots evaluated closed-loop tracking tasks, including echelon and column formation flight and target following. Cooper-Harper ratings and pilot comments were collected for each maneuver. This paper describes the test system, including the PSFCCs, the Gripen centerstick, and the flight test experiment. The paper presents results of longitudinal handling qualities maneuvers, including low order equivalent systems, Neal-Smith, and controls anticipation parameter analyses. The experiment showed that, while the centerstick controller provided a different aircraft feel, few handling qualities deficiencies resulted. It also demonstrated that the PSFCCs were useful for this kind of investigation.

  12. One-step (18)F-labeling of peptides for positron emission tomography imaging using the SiFA methodology.

    PubMed

    Wängler, Carmen; Niedermoser, Sabrina; Chin, Joshua; Orchowski, Katy; Schirrmacher, Esther; Jurkschat, Klaus; Iovkova-Berends, Liuba; Kostikov, Alexey P; Schirrmacher, Ralf; Wängler, Björn

    2012-11-01

    Here we present a procedure to label peptides with the positron-emitting radioisotope fluorine-18 ((18)F) using the silicon-fluoride acceptor (SiFA) labeling methodology. Positron emission tomography (PET) has gained high importance in noninvasive imaging of various diseases over the past decades, and thus new specific imaging probes for PET imaging, especially those labeled with (18)F, because of the advantageous properties of this nuclide, are highly sought after. N-terminally SiFA-modified peptides can be labeled with (18)F(-) in one step at room temperature (20-25 °C) or below without forming side products, thereby producing satisfactory radiochemical yields of 46 ± 1.5% (n = 10). The degree of chemoselectivity of the (18)F-introduction, which is based on simple isotopic exchange, allows for a facile cartridge-based purification fully devoid of HPLC implementation, thereby yielding peptides with specific activities between 44.4 and 62.9 GBq μmol(-1) (1,200-1,700 Ci mmol(-1)) within 25 min. PMID:23037309

  13. Multiaxis thrust vectoring using axisymmetric nozzles and postexit vanes on an F/A-18 configuration vehicle

    NASA Technical Reports Server (NTRS)

    Bowers, Albion H.; Noffz, Gregory K.; Grafton, Sue B.; Mason, Mary L.; Peron, Lee R.

    1991-01-01

    A ground-based investigation was conducted on an operational system of multiaxis thrust vectoring using postexit vanes around an axisymmetric nozzle. This thrust vectoring system will be tested on the NASA F/A-18 High Alpha Research Vehicle (HARV) aircraft. The system provides thrust vectoring capability in both pitch and yaw. Ground based data were gathered from two separate tests at NASA Langley Research Center. The first was a static test in the 16-foot Transonic Tunnel Cold-Jet Facility with a 14.25 percent scale model of the axisymmetric nozzle and the postexit vanes. The second test was conducted in the 30 by 60 foot wind tunnel with a 16 percent F/A-18 complete configuration model. Data from the two sets are being used to develop models of jet plume deflection and thrust loss as a function of vane deflection. In addition, an aerodynamic interaction model based on plume deflection angles will be developed. Results from the scale model nozzle test showed that increased vane deflection caused exhaust plume turning. Aerodynamic interaction effects consisted primarily of favorable interaction of moments and unfavorable interaction of forces caused by the vectored jet plume.

  14. A new clustering strategy

    NASA Astrophysics Data System (ADS)

    Feng, Jian-xin; Tang, Jia-fu; Wang, Guang-xing

    2007-04-01

    On the basis of the analysis of clustering algorithm that had been proposed for MANET, a novel clustering strategy was proposed in this paper. With the trust defined by statistical hypothesis in probability theory and the cluster head selected by node trust and node mobility, this strategy can realize the function of the malicious nodes detection which was neglected by other clustering algorithms and overcome the deficiency of being incapable of implementing the relative mobility metric of corresponding nodes in the MOBIC algorithm caused by the fact that the receiving power of two consecutive HELLO packet cannot be measured. It's an effective solution to cluster MANET securely.

  15. The Heliothis virescens cadherin protein expressed in Drosophila S2 cells functions as a receptor for Bacillus thuringiensis Cry1A but not Cry1Fa toxins.

    PubMed

    Jurat-Fuentes, Juan Luis; Adang, Michael J

    2006-08-15

    Genetic knockout of the BtR4 gene encoding the Heliothis virescens cadherin-like protein (HevCaLP) is linked to resistance against Cry1Ac toxin from Bacillus thuringiensis. However, the functional Cry1Ac receptor role of this protein has not been established. We previously proposed HevCaLP as a shared binding site for B. thuringiensis (Bt) Cry1A and Cry1Fa toxins in the midgut epithelium of H. virescens larvae. Considering that Cry1Ac and Cry1Fa are coexpressed in second-generation transgenic cotton for enhanced control of Heliothine and Spodoptera species, our model suggests the possibility of evolution of cross resistance via alteration of HevCaLP. To test whether HevCaLP is a Cry1Ac and Cry1Fa receptor, HevCaLP was transiently expressed on the surface of Drosophila melanogaster Schneider 2 (S2) cells. Expressed HevCaLP bound [(125)I]Cry1A toxins under native (dot blot) and denaturing (ligand blot) conditions. Affinity pull-down assays demonstrated that Cry1Fa does not bind to HevCaLP expressed in S2 cells or in solubilized brush border membrane proteins. Using a fluorescence-based approach, we tested the ability of expressed HevCaLP to mediate toxicity of Cry1A and Cry1Fa toxins. Cry1A toxins killed S2 cells expressing HevCaLP, whereas Cry1Fa toxin did not. Our results demonstrate that HevCaLP is a functional Cry1A but not Cry1Fa receptor. PMID:16893170

  16. Testing the CaR-FA-X Model: Investigating the Mechanisms Underlying Reduced Autobiographical Memory Specificity in Individuals With and Without a History of Depression

    PubMed Central

    Sumner, Jennifer A.; Mineka, Susan; Adam, Emma K.; Craske, Michelle G.; Vrshek-Schallhorn, Suzanne; Wolitzky-Taylor, Kate; Zinbarg, Richard E.

    2014-01-01

    Reduced autobiographical memory specificity (AMS) is an important cognitive phenomenon in major depressive disorder (MDD), but knowledge is lacking about its mechanisms. The CaR-FA-X model of Williams and colleagues proposed that three processes contribute to reduced AMS, alone or in interaction: capture and rumination (CaR), functional avoidance (FA), and impaired executive control (X). However, the entire CaR-FA-X model has not been tested. We addressed this gap in the literature by investigating contributions of the CaR-FA-X mechanisms to reduced AMS, alone or in interaction, in a subset of young adults (N=439) from the Northwestern-UCLA Youth Emotion Project. Participants were classified as those with (n=164) and without (n=275) a history of MDD at AMS assessment. They completed measures of: AMS; rumination (the brooding factor; CaR); childhood, adolescent, and early adulthood adversity (FA); avoidant coping (FA); and verbal fluency (X). Using structural equation modeling, we found greatest support for associations between reduced AMS and the capture and rumination, and impaired executive control mechanisms. In those with and without a history of MDD, brooding and verbal fluency interacted to contribute to reduced AMS. For participants without a history of MDD, lower verbal fluency (indicating impaired executive control) was associated with reduced AMS among those high on brooding. For participants with a history of MDD, lower verbal fluency was associated with reduced AMS among those low on brooding. The first finding was consistent with predictions from the CaR-FA-X model but the latter was not. Implications for conceptualizations of reduced AMS and its mechanisms are discussed. PMID:24978693

  17. Unconventional methods for clustering

    NASA Astrophysics Data System (ADS)

    Kotyrba, Martin

    2016-06-01

    Cluster analysis or clustering is a task of grouping a set of objects in such a way that objects in the same group (called a cluster) are more similar (in some sense or another) to each other than to those in other groups (clusters). It is the main task of exploratory data mining and a common technique for statistical data analysis used in many fields, including machine learning, pattern recognition, image analysis, information retrieval, and bioinformatics. The topic of this paper is one of the modern methods of clustering namely SOM (Self Organising Map). The paper describes the theory needed to understand the principle of clustering and descriptions of algorithm used with clustering in our experiments.

  18. Interaction of Boron Clusters with Oxygen: a DFT Study

    NASA Astrophysics Data System (ADS)

    Salavitabar, Kamron; Boggavarapu, Kiran; Kandalam, Anil

    A controlled combustion involving aluminum nanoparticles has often been the focus of studies in the field of solid fuel propellants. However very little focus has been given to the study of boron nanoparticles in controlled combustion. In contrast to aluminum nanoclusters, boron nanoclusters (Bn) are known to exhibit a planar geometries even at the size of n = 19 - 20, and thus offer a greater surface area for interaction with oxygen. Earlier experimental studies have shown that boron nanoclusters exhibit different reactivity with oxygen depending on their size and charge. In this poster, we present our recent density functional theory based results, focusing on the reactivity patterns of neutral and negatively charged B5 cluster with On, where n = 1 - 5; and B6 cluster with On (n = 1 - 2). The effect of charge on the reactivity of boron cluster, variation in the stability of product clusters, i e., neutral and negatively charged B5On (n = 1 - 5) and B6On (n = 1 - 2) are also examined. Financial Support from West Chester University Foundation under FaStR grant is acknowledged.

  19. Modeling Clustered Data with Very Few Clusters.

    PubMed

    McNeish, Daniel; Stapleton, Laura M

    2016-01-01

    Small-sample inference with clustered data has received increased attention recently in the methodological literature, with several simulation studies being presented on the small-sample behavior of many methods. However, nearly all previous studies focus on a single class of methods (e.g., only multilevel models, only corrections to sandwich estimators), and the differential performance of various methods that can be implemented to accommodate clustered data with very few clusters is largely unknown, potentially due to the rigid disciplinary preferences. Furthermore, a majority of these studies focus on scenarios with 15 or more clusters and feature unrealistically simple data-generation models with very few predictors. This article, motivated by an applied educational psychology cluster randomized trial, presents a simulation study that simultaneously addresses the extreme small sample and differential performance (estimation bias, Type I error rates, and relative power) of 12 methods to account for clustered data with a model that features a more realistic number of predictors. The motivating data are then modeled with each method, and results are compared. Results show that generalized estimating equations perform poorly; the choice of Bayesian prior distributions affects performance; and fixed effect models perform quite well. Limitations and implications for applications are also discussed. PMID:27269278

  20. [Clustering of simple obesity].

    PubMed

    Yoshida, K; Matsuda, H; Kurita, M; Umetada, Y

    1988-05-01

    An attempt was made to classify persons with simple obesity from the viewpoint of health education. Subjects of the study were 1,278 male workers in a financing company who underwent health examination. At the time of health examinations, questionnaire survey concerning their life styles was carried out on all the subjects. The obese group consisted of 127 subjects whose obesity indices were over 15% and the control group consisted of 342 subjects whose obesity indices ranged from -5 to 5%. Subjects in the obese group were classified into four clusters based on cluster analysis using five life-style parameters; that is, frequency of taking breakfast, frequency of taking staple food, drinking habits, smoking habits, and frequency of exercise. The first cluster (N = 10) included inactive persons, the second cluster (N = 46) non smokers, the third cluster (N = 39) smokers and heavy drinkers, and the fourth cluster (N = 32) smokers and non-drinkers. Comparison of the four clusters of obese persons with the control group revealed the following findings: 1) All the four clusters had significantly high frequencies of abnormal values of triglyceride (TG) and fasting blood sugar (FBS). 2) The first cluster had significantly high frequencies of abnormal values of glutamic oxalacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT). 3) The second cluster had significantly high frequencies of abnormal values of systolic and diastolic blood pressure, total cholesterol, TG, FBS, uric acid, GOT, GPT and gamma glutamyl transferase (GGT).(ABSTRACT TRUNCATED AT 250 WORDS) PMID:3172544

  1. Geophysical data collected from the St. Clair River between Michigan and Ontario, Canada (2008-016-FA)

    USGS Publications Warehouse

    Denny, Jane F.; Foster, D.S.; Worley, C.R.; Irwin, Barry J.

    2011-01-01

    In 2008, the U.S. Geological Survey (USGS), Woods Hole Coastal and Marine Science Center (WHCMSC), in cooperation with the U.S. Army Corps of Engineers conducted a geophysical and sampling survey of the riverbed of the Upper St. Clair River between Port Huron, Mich., and Sarnia, Ontario, Canada. The objectives were to define the Quaternary geologic framework of the riverbed of the St. Clair River to evaluate the relationship between morphologic change of the riverbed and underlying stratigraphy. This report presents the geophysical and sample data collected from the St. Clair River, May 29-June 6, 2008, as part of the International Upper Great Lakes Study, a 5-year project funded by the International Joint Commission of the United States and Canada to examine whether physical changes in the St. Clair River are affecting water levels within upper Great Lakes, to assess regulation plans for outflows from Lake Superior, and to examine the potential effect of climate change on the Great Lakes water levels (http://www.iugls.org). This document makes available the data that were used in a separate report, U.S. Geological Survey Open-File Report 2009-1137, which detailed the interpretations of the Quaternary geologic framework of the region. This report includes a description of the high-resolution acoustic and sediment-sampling systems that were used to map the morphology, surficial sediment distribution, and underlying geology of the Upper St. Clair River during USGS field activity 2008-016-FA (http://quashnet.er.usgs.gov/cgi-bin/datasource/public_ds_info.pl?fa=2008-016-FA). Video and photographs of the riverbed were also collected and are included in this data release. Future analyses will be focused on substrate erosion and its effects on river-channel morphology and geometry. Ultimately, the International Upper Great Lakes Study will attempt to determine where physical changes in the St. Clair River affect water flow and, subsequently, water levels in the Upper Great

  2. Electron: Cluster interactions

    SciTech Connect

    Scheidemann, A.A.; Kresin, V.V.; Knight, W.D.

    1994-02-01

    Beam depletion spectroscopy has been used to measure absolute total inelastic electron-sodium cluster collision cross sections in the energy range from E {approximately} 0.1 to E {approximately} 6 eV. The investigation focused on the closed shell clusters Na{sub 8}, Na{sub 20}, Na{sub 40}. The measured cross sections show an increase for the lowest collision energies where electron attachment is the primary scattering channel. The electron attachment cross section can be understood in terms of Langevin scattering, connecting this measurement with the polarizability of the cluster. For energies above the dissociation energy the measured electron-cluster cross section is energy independent, thus defining an electron-cluster interaction range. This interaction range increases with the cluster size.

  3. Information-based clustering

    PubMed Central

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

    2005-01-01

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

  4. Directionality of Electron Transfer in Type I Reaction Center Proteins: High-Frequency EPR Study of PS I with Removed Iron-Sulfur Centers.

    PubMed

    Poluektov, Oleg G; Utschig, Lisa M

    2015-10-29

    A key step of photosynthetic solar energy conversion involves rapid light-induced sequential electron-transfer steps that result in the formation of a stabilized charge-separated state. These primary reactions take place in large integral membrane reaction center (RC) proteins, wherein a series of donor/acceptor cofactors are specifically positioned for efficient electron transfer. RCs can be divided in two classes, Type I and Type II and examples of both types, photosystem I (PS I) and photosystem II (PS II), are involved in the oxygenic photosynthesis of higher plants, cyanobacteria, and algae. High-resolution X-ray crystal structures reveal that PS I and PS II contain two nearly symmetric branches of redox cofactors, termed the A and B branches. While unidirectional ET along the A branch in Type II RCs is well established, there is still a debate of whether primary photochemistry in Type I RCs is unidirectional along the A branch or bidirectional proceeding down both of the A and B branches. Light-induced electron transfer through the B branch has been observed in genetically modified PS I and in native PS I pretreated with strong reducing conditions to reduce three [4Fe-4S] clusters, the terminal electron acceptors of PS I; however, the extent of asymmetry of ET along both cofactor branches remains an open question. To prove that bidirectional ET in PS I is not simply an artifact of a reducing environment or genetic modification and to determine the degree of PS I ET asymmetry, we have examined biochemically modified Synechococcus leopoliensis PS I RCs, wherein the [4Fe-4S] clusters FX, FA, and FB have been removed to prevent secondary ET from phylloquinones (A1A/A1B) to FX. For these Fe-removed proteins, we observe that ET along both the A and B branches occurs with a ratio close to 1. Together with previously reported data, the concomitant structural and kinetic information obtained with HF EPR unambiguously proves the bidirectional nature of ET in PS I over

  5. Design and development of an F/A-18 inlet distortion rake: A cost and time saving solution

    NASA Technical Reports Server (NTRS)

    Yuhas, Andrew J.; Ray, Ronald J.; Burley, Richard R.; Steenken, William G.; Lechtenberg, Leon; Thornton, Don

    1994-01-01

    An innovative inlet total-pressure distortion measurement rake has been designed and developed for the F/A-18 A/B/C/D aircraft inlet. The design was conceived by NASA and General Electric Aircraft Engines (Evendale, Ohio). This rake has been flight qualified and flown in the F-18 High Alpha Research Vehicle (HARV) at NASA Dryden Flight Research Center. The rake's eight-legged, one-piece wagon wheel design was developed at a reduced cost and offers reduced installation time compared with traditional designs. The rake features 40 dual measurement ports for both low- and high-frequency pressure measurements with the high-frequency transducer mounted at the port. The high-frequency transducer offers direct absolute pressure measurements from low frequency to the highest frequency of interest, thereby allowing the rake to be used during highly dynamic aircraft maneuvers. Outstanding structural characteristics are inherent to the design through its construction and use of lightweight materials.

  6. Façade Greening: High-rise apartment building in Milan using pre-stressed concrete slab

    NASA Astrophysics Data System (ADS)

    Sun, Wenning; Li, Mingxin; Han, Yinong; Wang, Moqi; Ansourian, Peter

    2016-08-01

    In this project, one single level of the Façade Greening was designed and modelled using finite element method in Strand7. A static analysis was performed in order to understand the deflection and the stress due to the extra loads imposed by the soil and plants. The results produced by the linear static solver are compared with the strength of the materials and the European limitations. The maximum tension stress which exceeds the tensile strength in concrete is found in the root of the cantilever balcony. An alternative design of the cantilevered balcony with pre-stressed concrete slab is modelled separately for the balcony. Decrease is found in the tension stress and the significant improvement of deflection of the balcony with pre-stressed concrete slab. The dynamic loads such as wind and earthquake did not suggest significant effect on the pre-stressed concrete slab.

  7. Release and environmental impact of silver nanoparticles and conventional organic biocides from coated wooden façades.

    PubMed

    Künniger, Tina; Gerecke, Andreas C; Ulrich, Andrea; Huch, Anja; Vonbank, Roger; Heeb, Markus; Wichser, Adrian; Haag, Regula; Kunz, Petra; Faller, Markus

    2014-01-01

    This study represents for the first time a comprehensive assessment of functionality and environmental impacts of metallic silver nanoparticles (Ag-NP) compared to conventional organic biocides. Four different transparent, hydrophobic coatings of wooden outdoor façades were tested during one year outdoor weathering. The total silver release from products with Ag-NP was proportional to the overall erosion of the coating. The results indicate that the Ag-NPs are likely transformed to silver complexes, which are considerably less toxic than ionic silver. The protective effect of the silver containing coatings against mold, blue stain and algae was insufficient, even in immaculate and non-weathered conditions. The release of organic biocides from conventional coatings was dependent on the weather conditions, the type of biocide and the use in the base or top coat. The conventional coating showed a good overall performance free from mold, blue stain and algae until the end of the test period. PMID:24121422

  8. New approach for the identification and validation of a nonlinear F/A-18 model by use of neural networks.

    PubMed

    Boely, Nicolas; Botez, Ruxandra Mihaela

    2010-11-01

    This paper presents a new approach for identifying and validating the F/A-18 aeroservoelastic model, based on flight flutter tests. The neural network (NN), trained with five different flight flutter cases, is validated using 11 other flight flutter test (FFT) data. A total of 16 FFT cases were obtained for all three flight regimes (subsonic, transonic, and supersonic) at Mach numbers ranging between 0.85 and 1.30 and at altitudes of between 5000 and 25 000 ft. The results obtained highlight the efficiency of the multilayer perceptron NN in model identification. Optimization of the NN requires mixing of two proprieties: the hidden layer size reduction and four-layered NN performances. This paper shows that a four-layer NN with only 16 neurons is enough to create an accurate model. The fit coefficients were higher than 92% for both the identification and the validation test data, thus demonstrating accuracy of the NN. PMID:20875971

  9. Water tunnel flow visualization and wind tunnel data analysis of the F/A-18. [leading edge extension vortex effects

    NASA Technical Reports Server (NTRS)

    Erickson, G. E.

    1982-01-01

    Six degree of freedom studies were utilized to extract a band of yawing and rolling moment coefficients from the F/A-18 aircraft flight records. These were compared with 0.06 scale model data obtained in a 16T wind tunnel facility. The results, indicate the flight test yawing moment data exhibit an improvement over the wind tunnel data to near neutral stability and a significant reduction in lateral stability (again to anear neutral level). These data are consistent with the flight test results since the motion was characterized by a relatively slo departure. Flight tests repeated the slow yaw departure at M 0.3. Only 0.16 scale model wind tunnel data showed levels of lateral stability similar to the flight test results. Accordingly, geometric modifications were investigated on the 0.16 scale model in the 30x60 foot wind tunnel to improve high angle of attack lateral stability.

  10. Capability Description for NASA's F/A-18 TN 853 as a Testbed for the Integrated Resilient Aircraft Control Project

    NASA Technical Reports Server (NTRS)

    Hanson, Curt

    2009-01-01

    The NASA F/A-18 tail number (TN) 853 full-scale Integrated Resilient Aircraft Control (IRAC) testbed has been designed with a full array of capabilities in support of the Aviation Safety Program. Highlights of the system's capabilities include: 1) a quad-redundant research flight control system for safely interfacing controls experiments to the aircraft's control surfaces; 2) a dual-redundant airborne research test system for hosting multi-disciplinary state-of-the-art adaptive control experiments; 3) a robust reversionary configuration for recovery from unusual attitudes and configurations; 4) significant research instrumentation, particularly in the area of static loads; 5) extensive facilities for experiment simulation, data logging, real-time monitoring and post-flight analysis capabilities; and 6) significant growth capability in terms of interfaces and processing power.

  11. Electrical resistivity structure of the Valu Fa Ridge, Lau Basin, from marine controlled-source electromagnetic sounding

    NASA Astrophysics Data System (ADS)

    MacGregor, Lucy; Sinha, Martin; Constable, Steven

    2001-07-01

    In December 1995 we carried out a comprehensive controlled-source electromagnetic survey of the Valu Fa Ridge at 22°25'S in the Lau Basin. The Valu Fa Ridge is a back-arc spreading centre of intermediate spreading rate and is a site of extensive hydrothermal activity. Seismic studies have imaged a melt lens at an average depth of 3.2km below the seafloor, surrounded by a zone of lowered seismic velocity, interpreted as a region of partial melt in the crust. The electromagnetic experiment was part of a multidisciplinary study which included wide-angle and reflection seismics, bathymetry and potential field measurements. Electromagnetic signals at frequencies between 0.25 and 40Hz were transmitted from a horizontal electric dipole towed close to the seafloor and were recorded by an array of 11 sea-bottom receivers at ranges of up to 20km from the source. Over 80 hr of data, consisting of the magnitude of the horizontal electric field at the seafloor, were collected. These data have extremely low scatter compared to similar data from previous surveys. The data were interpreted using a combination of 1- and 2-D forward modelling and inversion. The vertical resistivity gradient in the upper crust at the Valu Fa Ridge is abnormally low, with resistivities of less than 10Ω m observed throughout layer 2 of the crust to a depth of 3km. This is significantly more conductive at depth than the axis of the slow-spreading Reykjanes Ridge at 57°45'N, and the fast-spreading East Pacific Rise at 13°N, where similar data sets have been collected in the past. Although the structure of layer 2 is well constrained by the electromagnetic data, its extremely low resistivity causes rapid attenuation of electromagnetic signals diffusing through it, and hence the data are not sensitive to the structure in layer 3, in particular the structure of the melt lens or surrounding low-velocity zone. The seismic velocity structure of the Valu Fa Ridge, determined from the coincident wide

  12. Mini-clusters

    NASA Technical Reports Server (NTRS)

    Chinellato, J. A.; Dobrigkeit, C.; Bellandifilho, J.; Lattes, C. M. G.; Menon, M. J.; Navia, C. E.; Pamilaju, A.; Sawayanagi, K.; Shibuya, E. H.; Turtelli, A., Jr.

    1985-01-01

    Experimental results of mini-clusters observed in Chacaltaya emulsion chamber no.19 are summarized. The study was made on 54 single core shower upper and 91 shower clusters of E(gamma) 10 TeV from 30 families which are visible energy greater than 80 TeV and penetrate through both upper and lower detectors of the two-story chamber. The association of hadrons in mini-cluster is made clear from their penetrative nature and microscopic observation of shower continuation in lower chamber. Small P sub t (gamma) of hadrons in mini-clusters remained in puzzle.

  13. Management of cluster headache.

    PubMed

    Tfelt-Hansen, Peer C; Jensen, Rigmor H

    2012-07-01

    The prevalence of cluster headache is 0.1% and cluster headache is often not diagnosed or misdiagnosed as migraine or sinusitis. In cluster headache there is often a considerable diagnostic delay - an average of 7 years in a population-based survey. Cluster headache is characterized by very severe or severe orbital or periorbital pain with a duration of 15-180 minutes. The cluster headache attacks are accompanied by characteristic associated unilateral symptoms such as tearing, nasal congestion and/or rhinorrhoea, eyelid oedema, miosis and/or ptosis. In addition, there is a sense of restlessness and agitation. Patients may have up to eight attacks per day. Episodic cluster headache (ECH) occurs in clusters of weeks to months duration, whereas chronic cluster headache (CCH) attacks occur for more than 1 year without remissions. Management of cluster headache is divided into acute attack treatment and prophylactic treatment. In ECH and CCH the attacks can be treated with oxygen (12 L/min) or subcutaneous sumatriptan 6 mg. For both oxygen and sumatriptan there are two randomized, placebo-controlled trials demonstrating efficacy. In both ECH and CCH, verapamil is the prophylactic drug of choice. Verapamil 360 mg/day was found to be superior to placebo in one clinical trial. In clinical practice, daily doses of 480-720 mg are mostly used. Thus, the dose of verapamil used in cluster headache treatment may be double the dose used in cardiology, and with the higher doses the PR interval should be checked with an ECG. At the start of a cluster, transitional preventive treatment such as corticosteroids or greater occipital nerve blockade can be given. In CCH and in long-standing clusters of ECH, lithium, methysergide, topiramate, valproic acid and ergotamine tartrate can be used as add-on prophylactic treatment. In drug-resistant CCH, neuromodulation with either occipital nerve stimulation or deep brain stimulation of the hypothalamus is an alternative treatment strategy

  14. The youngest globular clusters

    NASA Astrophysics Data System (ADS)

    Beck, Sara

    2015-11-01

    It is likely that all stars are born in clusters, but most clusters are not bound and disperse. None of the many protoclusters in our Galaxy are likely to develop into long-lived bound clusters. The super star clusters (SSCs) seen in starburst galaxies are more massive and compact and have better chances of survival. The birth and early development of SSCs takes place deep in molecular clouds, and during this crucial stage the embedded clusters are invisible to optical or UV observations but are studied via the radio-infrared supernebulae (RISN) they excite. We review observations of embedded clusters and identify RISN within 10 Mpc whose exciting clusters have ≈ 106 M⊙ or more in volumes of a few pc3 and which are likely to not only survive as bound clusters, but to evolve into objects as massive and compact as Galactic globulars. These clusters are distinguished by very high star formation efficiency η, at least a factor of 10 higher than the few percent seen in the Galaxy, probably due to the violent disturbances their host galaxies have undergone. We review recent observations of the kinematics of the ionized gas in RISN showing outflows through low-density channels in the ambient molecular cloud; this may protect the cloud from feedback by the embedded H II region.

  15. Production of Sophorolipid from an Identified Current Yeast, Lachancea thermotolerans BBMCZ7FA20, Isolated from Honey Bee.

    PubMed

    Mousavi, Fereshteh; Beheshti-Maal, Keivan; Massah, Ahmadreza

    2015-08-01

    Biosurfactants are a family of diverse amphipathic molecules that are produced by several microorganisms such as bacteria, molds, and yeasts. These surface active agents have several applications in agriculture, oil processing, food, and pharmaceutical industries. In this research using YMG and YUG culture media, a native yeast strain, HG5, was isolated from honey bee. The oil spread test as a screening method was used to evaluate biosurfactant production by the yeast HG5 isolate. The 5.8s-rDNA analysis confirmed that the isolated yeast was related to Lachancea thermotolerans. We named this strain Lachancea thermotolerans strain BBMCZ7FA20 and its 5.8s-rDNA sequence was deposited in GenBank, NCBI under accession number of KM042082.1. The best precursor of biosurfactant production was canola oil and the sophorolipid amount was measured for 24.2 g/l. The thin layer chromatography and Fourier Transform Infrared Spectroscopy analysis showed that the extracted biosurfactant from Lachancea thermotolerans was sophorolipid. In conclusion, this is the first report of sophorolipid production by a native yeast Lachancea thermotolerans BBMCZ7FA20 we isolated from the honey bee gut collected from an apiary farm in Saman, Chaharmahal Bakhtiari province, Iran. We suggested that some cost-effective supplements such as canola oil, sunflower oil, and corn oils could be applied for increasing the sophorolipid production by this native yeast strain. According to several applications of biosurfactants in today world, the production of sophorolipid by Lachancea thermotolerans could be considered as a potential in the current industrial microbiology and modern microbial biotechnology. PMID:26048484

  16. Antisense down-regulation of the strawberry β-galactosidase gene FaβGal4 increases cell wall galactose levels and reduces fruit softening

    PubMed Central

    Paniagua, Candelas; Blanco-Portales, Rosario; Barceló-Muñoz, Marta; García-Gago, Juan A.; Waldron, Keith W.; Quesada, Miguel A.; Muñoz-Blanco, Juan; Mercado, José A.

    2016-01-01

    Strawberry softening is characterized by an increase in the solubilization and depolymerization of pectins from cell walls. Galactose release from pectin side chains by β-galactosidase enzymes has been proposed as one reason for the increase in soluble pectins. A putative β-galactosidase gene, FaβGal4, has been identified using a custom-made oligonucleotide-based strawberry microarray platform. FaβGal4 was expressed mainly in the receptacle during fruit ripening, and was positively regulated by abscisic acid and negatively regulated by auxins. To ascertain the role of FaβGal4 in strawberry softening, transgenic plants containing an antisense sequence of this gene under the control of the CaMV35S promoter were generated. Phenotypic analyses were carried out in transgenic plants during three consecutive growing seasons, using non-transformed plants as control. Two out of nine independent transgenic lines yielded fruits that were 30% firmer than control at the ripe stage. FaβGal4 mRNA levels were reduced by 70% in ripe fruits from these selected transgenic lines, but they also showed significant silencing of FaβGal1, although the genes did not share significant similarity. These two transgenic lines also showed an increase in pectin covalently bound to the cell wall, extracted using Na2CO3. The amount of galactose in cell walls from transgenic fruits was 30% higher than in control; notably, the galactose increase was larger in the 1 M KOH fraction, which is enriched in hemicellulose. These results suggest that FaβGal4 participates in the solubilization of covalently bound pectins during ripening, reducing strawberry fruit firmness. PMID:26585222

  17. FaQR, Required for the Biosynthesis of the Strawberry Flavor Compound 4-Hydroxy-2,5-Dimethyl-3(2H)-Furanone, Encodes an Enone Oxidoreductase

    PubMed Central

    Raab, Thomas; López-Ráez, Juan Antonio; Klein, Dorothée; Caballero, Jose Luis; Moyano, Enriqueta; Schwab, Wilfried; Muñoz-Blanco, Juan

    2006-01-01

    The flavor of strawberry (Fragaria × ananassa) fruit is dominated by an uncommon group of aroma compounds with a 2,5-dimethyl-3(H)-furanone structure. We report the characterization of an enzyme involved in the biosynthesis of 4-hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF; Furaneol), the key flavor compound in strawberries. Protein extracts were partially purified, and the observed distribution of enzymatic activity correlated with the presence of a single polypeptide of ∼37 kD. Sequence analysis of two peptide fragments showed total identity with the protein sequence of a strongly ripening-induced, auxin-dependent putative quinone oxidoreductase, Fragaria × ananassa quinone oxidoreductase (FaQR). The open reading frame of the FaQR cDNA consists of 969 bp encoding a 322–amino acid protein with a calculated molecular mass of 34.3 kD. Laser capture microdissection followed by RNA extraction and amplification demonstrated the presence of FaQR mRNA in parenchyma tissue of the strawberry fruit. The FaQR protein was functionally expressed in Escherichia coli, and the monomer catalyzed the formation of HDMF. After chemical synthesis and liquid chromatography–tandem mass spectrometry analysis, 4-hydroxy-5-methyl-2-methylene-3(2H)-furanone was confirmed as a substrate of FaQR and the natural precursor of HDMF. This study demonstrates the function of the FaQR enzyme in the biosynthesis of HDMF as enone oxidoreductase and provides a foundation for the improvement of strawberry flavor and the biotechnological production of HDMF. PMID:16517758

  18. FaQR, required for the biosynthesis of the strawberry flavor compound 4-hydroxy-2,5-dimethyl-3(2H)-furanone, encodes an enone oxidoreductase.

    PubMed

    Raab, Thomas; López-Ráez, Juan Antonio; Klein, Dorothée; Caballero, Jose Luis; Moyano, Enriqueta; Schwab, Wilfried; Muñoz-Blanco, Juan

    2006-04-01

    The flavor of strawberry (Fragaria x ananassa) fruit is dominated by an uncommon group of aroma compounds with a 2,5-dimethyl-3(H)-furanone structure. We report the characterization of an enzyme involved in the biosynthesis of 4-hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF; Furaneol), the key flavor compound in strawberries. Protein extracts were partially purified, and the observed distribution of enzymatic activity correlated with the presence of a single polypeptide of approximately 37 kD. Sequence analysis of two peptide fragments showed total identity with the protein sequence of a strongly ripening-induced, auxin-dependent putative quinone oxidoreductase, Fragaria x ananassa quinone oxidoreductase (FaQR). The open reading frame of the FaQR cDNA consists of 969 bp encoding a 322-amino acid protein with a calculated molecular mass of 34.3 kD. Laser capture microdissection followed by RNA extraction and amplification demonstrated the presence of FaQR mRNA in parenchyma tissue of the strawberry fruit. The FaQR protein was functionally expressed in Escherichia coli, and the monomer catalyzed the formation of HDMF. After chemical synthesis and liquid chromatography-tandem mass spectrometry analysis, 4-hydroxy-5-methyl-2-methylene-3(2H)-furanone was confirmed as a substrate of FaQR and the natural precursor of HDMF. This study demonstrates the function of the FaQR enzyme in the biosynthesis of HDMF as enone oxidoreductase and provides a foundation for the improvement of strawberry flavor and the biotechnological production of HDMF. PMID:16517758

  19. Antisense Down-Regulation of the FaPG1 Gene Reveals an Unexpected Central Role for Polygalacturonase in Strawberry Fruit Softening1[W

    PubMed Central

    Quesada, Miguel A.; Blanco-Portales, Rosario; Posé, Sara; García-Gago, Juan A.; Jiménez-Bermúdez, Silvia; Muñoz-Serrano, Andrés; Caballero, José L.; Pliego-Alfaro, Fernando; Mercado, José A.; Muñoz-Blanco, Juan

    2009-01-01

    The strawberry (Fragaria × ananassa ‘Chandler’) fruit undergoes a fast softening during ripening. Polygalacturonase (PG) activity is low during this process, but two ripening-related PG genes, FaPG1 and FaPG2, have been cloned. Both genes were up-regulated during fruit ripening and were also negatively regulated by auxin. To further assess the role of FaPG1 on strawberry softening, transgenic plants containing an antisense sequence of this gene under the control of the 35S promoter (APG lines) were obtained. Sixteen out of 30 independent transgenic lines showed fruit yields similar to those of the control. Several quality parameters were measured in ripe fruits from these 16 lines. Fruit weight was slightly reduced in four lines, and most of them showed an increase in soluble solid content. Half of these lines yielded fruits significantly firmer than did the control. Four APG lines were selected, their ripened fruits being on average 163% firmer than the control. The postharvest softening of APG fruits was also diminished. Ripened fruits from the four selected lines showed a 90% to 95% decrease in FaPG1 transcript abundance, whereas the level of FaPG2 was not significantly altered. Total PG activity was reduced in three of these lines when compared with control fruits. Cell wall extracts from APG fruits showed a reduction in pectin solubilization and an increase in pectins covalently bound to the cell wall. A comparative transcriptomic analysis of gene expression between the ripened receptacle of the control and those of the APG fruits (comprising 1,250 receptacle expressed sequence tags) did not show any statistically significant change. These results indicate that FaPG1 plays a central role in strawberry softening. PMID:19395408

  20. Antisense down-regulation of the strawberry β-galactosidase gene FaβGal4 increases cell wall galactose levels and reduces fruit softening.

    PubMed

    Paniagua, Candelas; Blanco-Portales, Rosario; Barceló-Muñoz, Marta; García-Gago, Juan A; Waldron, Keith W; Quesada, Miguel A; Muñoz-Blanco, Juan; Mercado, José A

    2016-02-01

    Strawberry softening is characterized by an increase in the solubilization and depolymerization of pectins from cell walls. Galactose release from pectin side chains by β-galactosidase enzymes has been proposed as one reason for the increase in soluble pectins. A putative β-galactosidase gene, FaβGal4, has been identified using a custom-made oligonucleotide-based strawberry microarray platform. FaβGal4 was expressed mainly in the receptacle during fruit ripening, and was positively regulated by abscisic acid and negatively regulated by auxins. To ascertain the role of FaβGal4 in strawberry softening, transgenic plants containing an antisense sequence of this gene under the control of the CaMV35S promoter were generated. Phenotypic analyses were carried out in transgenic plants during three consecutive growing seasons, using non-transformed plants as control. Two out of nine independent transgenic lines yielded fruits that were 30% firmer than control at the ripe stage. FaβGal4 mRNA levels were reduced by 70% in ripe fruits from these selected transgenic lines, but they also showed significant silencing of FaβGal1, although the genes did not share significant similarity. These two transgenic lines also showed an increase in pectin covalently bound to the cell wall, extracted using Na2CO3. The amount of galactose in cell walls from transgenic fruits was 30% higher than in control; notably, the galactose increase was larger in the 1 M KOH fraction, which is enriched in hemicellulose. These results suggest that FaβGal4 participates in the solubilization of covalently bound pectins during ripening, reducing strawberry fruit firmness. PMID:26585222

  1. The mannose-binding lectin gene FaMBL1 is involved in the resistance of unripe strawberry fruits to Colletotrichum acutatum.

    PubMed

    Guidarelli, Michela; Zoli, Lisa; Orlandini, Alessandro; Bertolini, Paolo; Baraldi, Elena

    2014-10-01

    The fungal pathogen Colletotrichum acutatum is the causal agent of strawberry (Fragaria × ananassa) anthracnose. Although the fungus can infect strawberry fruits at both unripe and ripe stages, the symptoms appear only on red ripe fruits. On white unripe fruits, the pathogen becomes quiescent as melanized appressoria after 24 h of interaction. Previous transcriptome analysis has indicated that a mannose-binding lectin (MBL) gene is the most up-regulated gene in 24-h-infected white strawberries, suggesting a role for this gene in the low susceptibility of unripe stages. A time course analysis of the expression of this MBL gene, named FaMBL1 (Fragaria × ananassa MBL 1a), was undertaken to monitor its expression profile in white and red fruits at early interaction times: FaMBL1 was expressed exclusively in white fruit after 24 h, when the pathogen was quiescent. Agrobacterium-mediated transient transformation was used to silence and overexpress the FaMBL1 gene in 24-h-infected white and red strawberries, respectively. FaMBL1-silenced unripe fruits showed an increase in susceptibility to C. acutatum. These 24-h-infected tissues contained subcuticular hyphae, indicating pathogen penetration and active growth. In contrast, overexpression of FaMBL1 in ripe fruits decreased susceptibility; here, 24-h-infected tissues showed a high percentage of ungerminated appressoria, suggesting that the growth of the pathogen had slowed. These data suggest that FaMBL1 plays a crucial role in the resistance of unripe strawberry fruits to C. acutatum. PMID:24690196

  2. Identification of differentially expressed genes associated with the enhancement of X-ray susceptibility by RITA in a hypopharyngeal squamous cell carcinoma cell line (FaDu)

    PubMed Central

    Luan, Jinwei; Li, Xianglan; Guo, Rutao; Liu, Shanshan; Luo, Hongyu

    2016-01-01

    Abstract Background Next generation sequencing and bio-informatic analyses were conducted to investigate the mechanism of reactivation of p53 and induction of tumor cell apoptosis (RITA)-enhancing X-ray susceptibility in FaDu cells. Materials and methods The cDNA was isolated from FaDu cells treated with 0 X-ray, 8 Gy X-ray, or 8 Gy X-ray + RITA. Then, cDNA libraries were created and sequenced using next generation sequencing, and each assay was repeated twice. Subsequently, differentially expressed genes (DEGs) were identified using Cuffdiff in Cufflinks and their functions were predicted by pathway enrichment analyses. Genes that were constantly up- or down-regulated in 8 Gy X-ray-treated FaDu cells and 8 Gy X-ray + RITA-treated FaDu cells were obtained as RITA genes. Afterward, the protein-protein interaction (PPI) relationships were obtained from the STRING database and a PPI network was constructed using Cytoscape. Furthermore, ClueGO was used for pathway enrichment analysis of genes in the PPI network. Results Total 2,040 and 297 DEGs were identified in FaDu cells treated with 8 Gy X-ray or 8 Gy X-ray + RITA, respectively. PARP3 and NEIL1 were enriched in base excision repair, and CDK1 was enriched in p53 signaling pathway. RFC2 and EZH2 were identified as RITA genes. In the PPI network, many interaction relationships were identified (e.g., RFC2-CDK1, EZH2-CDK1 and PARP3-EZH2). ClueGO analysis showed that RFC2 and EZH2 were related to cell cycle. Conclusions RFC2, EZH2, CDK1, PARP3 and NEIL1 may be associated, and together enhance the susceptibility of FaDu cells treated with RITA to the deleterious effects of X-ray. PMID:27247549

  3. Clustering versus non-clustering phase synchronizations.

    PubMed

    Liu, Shuai; Zhan, Meng

    2014-03-01

    Clustering phase synchronization (CPS) is a common scenario to the global phase synchronization of coupled dynamical systems. In this work, a novel scenario, the non-clustering phase synchronization (NPS), is reported. It is found that coupled systems do not transit to the global synchronization until a certain sufficiently large coupling is attained, and there is no clustering prior to the global synchronization. To reveal the relationship between CPS and NPS, we further analyze the noise effect on coupled phase oscillators and find that the coupled oscillator system can change from CPS to NPS with the increase of noise intensity or system disorder. These findings are expected to shed light on the mechanism of various intriguing self-organized behaviors in coupled systems. PMID:24697366

  4. Clustering versus non-clustering phase synchronizations

    SciTech Connect

    Liu, Shuai; Zhan, Meng

    2014-03-15

    Clustering phase synchronization (CPS) is a common scenario to the global phase synchronization of coupled dynamical systems. In this work, a novel scenario, the non-clustering phase synchronization (NPS), is reported. It is found that coupled systems do not transit to the global synchronization until a certain sufficiently large coupling is attained, and there is no clustering prior to the global synchronization. To reveal the relationship between CPS and NPS, we further analyze the noise effect on coupled phase oscillators and find that the coupled oscillator system can change from CPS to NPS with the increase of noise intensity or system disorder. These findings are expected to shed light on the mechanism of various intriguing self-organized behaviors in coupled systems.

  5. SPASM and Twitch Domains in S-Adenosylmethionine (SAM) Radical Enzymes*

    PubMed Central

    Grell, Tsehai A. J.; Goldman, Peter J.; Drennan, Catherine L.

    2015-01-01

    S-Adenosylmethionine (SAM, also known as AdoMet) radical enzymes use SAM and a [4Fe-4S] cluster to catalyze a diverse array of reactions. They adopt a partial triose-phosphate isomerase (TIM) barrel fold with N- and C-terminal extensions that tailor the structure of the enzyme to its specific function. One extension, termed a SPASM domain, binds two auxiliary [4Fe-4S] clusters and is present within peptide-modifying enzymes. The first structure of a SPASM-containing enzyme, anaerobic sulfatase-maturating enzyme (anSME), revealed unexpected similarities to two non-SPASM proteins, butirosin biosynthetic enzyme 2-deoxy-scyllo-inosamine dehydrogenase (BtrN) and molybdenum cofactor biosynthetic enzyme (MoaA). The latter two enzymes bind one auxiliary cluster and exhibit a partial SPASM motif, coined a Twitch domain. Here we review the structure and function of auxiliary cluster domains within the SAM radical enzyme superfamily. PMID:25477505

  6. A nonparametric clustering technique which estimates the number of clusters

    NASA Technical Reports Server (NTRS)

    Ramey, D. B.

    1983-01-01

    In applications of cluster analysis, one usually needs to determine the number of clusters, K, and the assignment of observations to each cluster. A clustering technique based on recursive application of a multivariate test of bimodality which automatically estimates both K and the cluster assignments is presented.

  7. Reactive accelerated cluster erosion (RACE) by ionized cluster beams

    NASA Astrophysics Data System (ADS)

    Gspann, Jürgen

    1996-05-01

    Beams of ionized clusters accelerated up to about 120 keV kinetic energy per cluster are used for cluster impact lithography. Chemical reactions of clusters of CO 2, or of SF 6, respectively, are found to assist the physical erosion by hypervelocity cluster impacts in yielding volatile products. Natural diamond, silicon and Pyrex glass have been microstructured showing very smooth eroded surfaces.

  8. In-Flight Suppression of a De-Stabilized F/A-18 Structural Mode Using the Space Launch System Adaptive Augmenting Control System

    NASA Technical Reports Server (NTRS)

    Wall, John; VanZwieten, Tannen; Giiligan Eric; Miller, Chris; Hanson, Curtis; Orr, Jeb

    2015-01-01

    Adaptive Augmenting Control (AAC) has been developed for NASA's Space Launch System (SLS) family of launch vehicles and implemented as a baseline part of its flight control system (FCS). To raise the technical readiness level of the SLS AAC algorithm, the Launch Vehicle Adaptive Control (LVAC) flight test program was conducted in which the SLS FCS prototype software was employed to control the pitch axis of Dryden's specially outfitted F/A-18, the Full Scale Advanced Systems Test Bed (FAST). This presentation focuses on a set of special test cases which demonstrate the successful mitigation of the unstable coupling of an F/A-18 airframe structural mode with the SLS FCS.

  9. Investigating potential planetary nebula/cluster pairs

    NASA Astrophysics Data System (ADS)

    Moni Bidin, C.; Majaess, D.; Bonatto, C.; Mauro, F.; Turner, D.; Geisler, D.; Chené, A.-N.; Gormaz-Matamala, A. C.; Borissova, J.; Kurtev, R. G.; Minniti, D.; Carraro, G.; Gieren, W.

    2014-01-01

    Context. Fundamental parameters characterizing the end-state of intermediate-mass stars may be constrained by discovering planetary nebulae (PNe) in open clusters (OCs). Cluster membership may be exploited to establish the distance, luminosity, age, and physical size for PNe, and the intrinsic luminosity and mass of its central star. Aims: Four potential PN-OC associations were investigated to assess the cluster membership for the PNe. Methods: Radial velocities were measured from intermediate-resolution optical spectra, complemented with previous estimates in the literature. When the radial velocity study supported the PN/OC association, we analyzed whether other parameters (e.g., age, distance, reddening, central star brightness) were consistent with this conclusion. Results: Our measurements imply that the PNe VBe 3 and HeFa 1 are not members of the OCs NGC 5999 and NGC 6067, respectively, and that they very likely belong to the background bulge population. Conversely, consistent radial velocities indicate that NGC 2452/NGC 2453 could be associated, but our results are not conclusive so additional observations are warranted. Finally, we demonstrate that all the available information point to He 2-86 being a young, highly internally obscured PN member of NGC 4463. New near-infrared photometry acquired via the Vista Variables in the Via Lactea ESO public survey was used in tandem with existing UBV photometry to measure the distance, reddening, and age of NGC 4463, finding d = 1.55 ± 0.10 kpc, E(B - V) = 0.41 ± 0.02, and τ = 65 ± 10 Myr, respectively. The same values should be adopted for the PN if the proposed cluster membership is confirmed. Based on observations gathered with ESO-VISTA telescope (program ID 172.B-2002).Based on observations gathered at Las Campanas observatory (program ID CN2012A-080).The spectra as FITS files are only available at the CDS via anonymous ftp to http://cdsarc.u-strasbg.fr (ftp://130.79.128.5) or via http://cdsarc

  10. The LLNL Cluster Tool

    SciTech Connect

    Hunter, S L

    2007-03-27

    {lg_bullet} The Cluster Tool -is a set of linked vacuum chambers -can deposit multiple layers of metal and metal oxides {lg_bullet} Each layer can be deposited without breaking vacuum {lg_bullet} Shadow masks can give each layer a different pattern {lg_bullet} The Cluster Tool will be operational in April

  11. Cluster Interest Inventory.

    ERIC Educational Resources Information Center

    Herzog, Douglas

    The Cluster Interest Inventory is designed to familiarize students with representative occupations in 13 career clusters: (1) agribusiness and natural resources, (2) business marketing, and office occupations, (3) communications and media, (4) consumer and homemaker, (5) fine arts and humanities, (6) health, (7) manufacturing and processing, (8)…

  12. Coma cluster of galaxies

    NASA Technical Reports Server (NTRS)

    1999-01-01

    Atlas Image mosaic, covering 34' x 34' on the sky, of the Coma cluster, aka Abell 1656. This is a particularly rich cluster of individual galaxies (over 1000 members), most prominently the two giant ellipticals, NGC 4874 (right) and NGC 4889 (left). The remaining members are mostly smaller ellipticals, but spiral galaxies are also evident in the 2MASS image. The cluster is seen toward the constellation Coma Berenices, but is actually at a distance of about 100 Mpc (330 million light years, or a redshift of 0.023) from us. At this distance, the cluster is in what is known as the 'Hubble flow,' or the overall expansion of the Universe. As such, astronomers can measure the Hubble Constant, or the universal expansion rate, based on the distance to this cluster. Large, rich clusters, such as Coma, allow astronomers to measure the 'missing mass,' i.e., the matter in the cluster that we cannot see, since it gravitationally influences the motions of the member galaxies within the cluster. The near-infrared maps the overall luminous mass content of the member galaxies, since the light at these wavelengths is dominated by the more numerous older stellar populations. Galaxies, as seen by 2MASS, look fairly smooth and homogeneous, as can be seen from the Hubble 'tuning fork' diagram of near-infrared galaxy morphology. Image mosaic by S. Van Dyk (IPAC).

  13. Probability and Cancer Clusters

    ERIC Educational Resources Information Center

    Hamilton-Keene, Rachael; Lenard, Christoper T.; Mills, Terry M.

    2009-01-01

    Recently there have been several news items about possible cancer clusters in the Australian media. The term "cancer cluster" is used when an unusually large number of people in one geographic area, often a workplace, are diagnosed with cancer in a short space of time. In this paper the authors explore this important health issue using probability…

  14. Illinois' Career Cluster Model

    ERIC Educational Resources Information Center

    Jankowski, Natasha A.; Kirby, Catherine L.; Bragg, Debra D.; Taylor, Jason L.; Oertle, Kathleen M.

    2009-01-01

    This booklet provides information to multiple stakeholders on the implementation of career clusters in Illinois. The booklet is an extension of the previous edition titled "An Introduction to Illinois CTE Programs of Study" (2008), and provides a resource for partners to understand Illinois' Career Cluster Model as its own adaptation of the…

  15. Matlab Cluster Ensemble Toolbox

    2009-04-27

    This is a Matlab toolbox for investigating the application of cluster ensembles to data classification, with the objective of improving the accuracy and/or speed of clustering. The toolbox divides the cluster ensemble problem into four areas, providing functionality for each. These include, (1) synthetic data generation, (2) clustering to generate individual data partitions and similarity matrices, (3) consensus function generation and final clustering to generate ensemble data partitioning, and (4) implementation of accuracy metrics. Withmore » regard to data generation, Gaussian data of arbitrary dimension can be generated. The kcenters algorithm can then be used to generate individual data partitions by either, (a) subsampling the data and clustering each subsample, or by (b) randomly initializing the algorithm and generating a clustering for each initialization. In either case an overall similarity matrix can be computed using a consensus function operating on the individual similarity matrices. A final clustering can be performed and performance metrics are provided for evaluation purposes.« less

  16. Mixed-Initiative Clustering

    ERIC Educational Resources Information Center

    Huang, Yifen

    2010-01-01

    Mixed-initiative clustering is a task where a user and a machine work collaboratively to analyze a large set of documents. We hypothesize that a user and a machine can both learn better clustering models through enriched communication and interactive learning from each other. The first contribution or this thesis is providing a framework of…

  17. Young Massive Star Clusters

    NASA Astrophysics Data System (ADS)

    Portegies Zwart, Simon F.; McMillan, Stephen L. W.; Gieles, Mark

    2010-09-01

    Young massive clusters (YMCs) are dense aggregates of young stars that form the fundamental building blocks of galaxies. Several examples exist in the Milky Way Galaxy and the Local Group, but they are particularly abundant in starburst and interacting galaxies. The few YMCs that are close enough to resolve are of prime interest for studying the stellar mass function and the ecological interplay between stellar evolution and stellar dynamics. The distant unresolved clusters may be effectively used to study the star-cluster mass function, and they provide excellent constraints on the formation mechanisms of young cluster populations. YMCs are expected to be the nurseries for many unusual objects, including a wide range of exotic stars and binaries. So far only a few such objects have been found in YMCs, although their older cousins, the globular clusters, are unusually rich in stellar exotica. In this review, we focus on star clusters younger than ˜100 Myr, more than a few current crossing times old, and more massive than ˜104M⊙; the size of the cluster and its environment are considered less relevant as distinguishing parameters. We describe the global properties of the currently known young massive star clusters in the Local Group and beyond, and discuss the state of the art in observations and dynamical modeling of these systems. In order to make this review readable by observers, theorists, and computational astrophysicists, we also review the cross-disciplinary terminology.

  18. Blue emitting undecaplatinum clusters

    NASA Astrophysics Data System (ADS)

    Chakraborty, Indranath; Bhuin, Radha Gobinda; Bhat, Shridevi; Pradeep, T.

    2014-07-01

    A blue luminescent 11-atom platinum cluster showing step-like optical features and the absence of plasmon absorption was synthesized. The cluster was purified using high performance liquid chromatography (HPLC). Electrospray ionization (ESI) and matrix assisted laser desorption ionization (MALDI) mass spectrometry (MS) suggest a composition, Pt11(BBS)8, which was confirmed by a range of other experimental tools. The cluster is highly stable and compatible with many organic solvents.A blue luminescent 11-atom platinum cluster showing step-like optical features and the absence of plasmon absorption was synthesized. The cluster was purified using high performance liquid chromatography (HPLC). Electrospray ionization (ESI) and matrix assisted laser desorption ionization (MALDI) mass spectrometry (MS) suggest a composition, Pt11(BBS)8, which was confirmed by a range of other experimental tools. The cluster is highly stable and compatible with many organic solvents. Electronic supplementary information (ESI) available: Details of experimental procedures, instrumentation, chromatogram of the crude cluster; SEM/EDAX, DLS, PXRD, TEM, FT-IR, and XPS of the isolated Pt11 cluster; UV/Vis, MALDI MS and SEM/EDAX of isolated 2 and 3; and 195Pt NMR of the K2PtCl6 standard. See DOI: 10.1039/c4nr02778g

  19. Brightest Cluster Galaxy Identification

    NASA Astrophysics Data System (ADS)

    Leisman, Luke; Haarsma, D. B.; Sebald, D. A.; ACCEPT Team

    2011-01-01

    Brightest cluster galaxies (BCGs) play an important role in several fields of astronomical research. The literature includes many different methods and criteria for identifying the BCG in the cluster, such as choosing the brightest galaxy, the galaxy nearest the X-ray peak, or the galaxy with the most extended profile. Here we examine a sample of 75 clusters from the Archive of Chandra Cluster Entropy Profile Tables (ACCEPT) and the Sloan Digital Sky Survey (SDSS), measuring masked magnitudes and profiles for BCG candidates in each cluster. We first identified galaxies by hand; in 15% of clusters at least one team member selected a different galaxy than the others.We also applied 6 other identification methods to the ACCEPT sample; in 30% of clusters at least one of these methods selected a different galaxy than the other methods. We then developed an algorithm that weighs brightness, profile, and proximity to the X-ray peak and centroid. This algorithm incorporates the advantages of by-hand identification (weighing multiple properties) and automated selection (repeatable and consistent). The BCG population chosen by the algorithm is more uniform in its properties than populations selected by other methods, particularly in the relation between absolute magnitude (a proxy for galaxy mass) and average gas temperature (a proxy for cluster mass). This work supported by a Barry M. Goldwater Scholarship and a Sid Jansma Summer Research Fellowship.

  20. Marketing Occupations. Cluster Guide.

    ERIC Educational Resources Information Center

    Oregon State Dept. of Education, Salem.

    This cluster guide, which is designed to show teachers what specific knowledge and skills qualify high school students for entry-level employment (or postsecondary training) in marketing occupations, is organized into three sections: (1) cluster organization and implementation, (2) instructional emphasis areas, and (3) assessment. The first…

  1. Muster: Massively Scalable Clustering

    2010-05-20

    Muster is a framework for scalable cluster analysis. It includes implementations of classic K-Medoids partitioning algorithms, as well as infrastructure for making these algorithms run scalably on very large systems. In particular, Muster contains algorithms such as CAPEK (described in reference 1) that are capable of clustering highly distributed data sets in-place on a hundred thousand or more processes.

  2. Structure and Function of 4-Hydroxyphenylacetate Decarboxylase and Its Cognate Activating Enzyme.

    PubMed

    Selvaraj, Brinda; Buckel, Wolfgang; Golding, Bernard T; Ullmann, G Matthias; Martins, Berta M

    2016-01-01

    4-Hydroxyphenylacetate decarboxylase (4Hpad) is the prototype of a new class of Fe-S cluster-dependent glycyl radical enzymes (Fe-S GREs) acting on aromatic compounds. The two-enzyme component system comprises a decarboxylase responsible for substrate conversion and a dedicated activating enzyme (4Hpad-AE). The decarboxylase uses a glycyl/thiyl radical dyad to convert 4-hydroxyphenylacetate into p-cresol (4-methylphenol) by a biologically unprecedented Kolbe-type decarboxylation. In addition to the radical dyad prosthetic group, the decarboxylase unit contains two [4Fe-4S] clusters coordinated by an extra small subunit of unknown function. 4Hpad-AE reductively cleaves S-adenosylmethionine (SAM or AdoMet) at a site-differentiated [4Fe-4S]2+/+ cluster (RS cluster) generating a transient 5'-deoxyadenosyl radical that produces a stable glycyl radical in the decarboxylase by the abstraction of a hydrogen atom. 4Hpad-AE binds up to two auxiliary [4Fe-4S] clusters coordinated by a ferredoxin-like insert that is C-terminal to the RS cluster-binding motif. The ferredoxin-like domain with its two auxiliary clusters is not vital for SAM-dependent glycyl radical formation in the decarboxylase, but facilitates a longer lifetime for the radical. This review describes the 4Hpad and cognate AE families and focuses on the recent advances and open questions concerning the structure, function and mechanism of this novel Fe-S-dependent class of GREs. PMID:26959876

  3. Cosmology with galaxy clusters

    NASA Astrophysics Data System (ADS)

    Sartoris, Barbara

    2015-08-01

    Clusters of galaxies are powerful probes to constrain parameters that describe the cosmological models and to distinguish among different models. Since, the evolution of the cluster mass function and large-scale clustering contain the informations about the linear growth rate of perturbations and the expansion history of the Universe, clusters have played an important role in establishing the current cosmological paradigm. It is crucial to know how to determine the cluster mass from observational quantities when using clusters as cosmological tools. For this, numerical simulations are helpful to define and study robust cluster mass proxies that have minimal and well understood scatter across the mass and redshift ranges of interest. Additionally, the bias in cluster mass determination can be constrained via observations of the strong and weak lensing effect, X-ray emission, the Sunyaev- Zel’dovic effect, and the dynamics of galaxies.A major advantage of X-ray surveys is that the observable-mass relation is tight. Moreover, clusters can be easily identified in X-ray as continuous, extended sources. As of today, interesting cosmological constraints have been obtained from relatively small cluster samples (~102), X-ray selected by the ROSAT satellite over a wide redshift range (0clusters, the ROSAT All-Sky Survey.The next generation of X-ray telescopes will enhance the statistics of detected clusters and enlarge their redshift coverage. In particular, eROSITA will produce a catalog of >105 clusters with photometric redshifts from multi-band optical surveys (e.g. PanSTARRS, DES, and LSST). This will vastly improve upon current cosmological constraints, especially by the synergy with other cluster surveys that

  4. Cool Cluster Correctly Correlated

    SciTech Connect

    Sergey Aleksandrovich Varganov

    2005-12-17

    Atomic clusters are unique objects, which occupy an intermediate position between atoms and condensed matter systems. For a long time it was thought that physical and chemical properties of atomic dusters monotonically change with increasing size of the cluster from a single atom to a condensed matter system. However, recently it has become clear that many properties of atomic clusters can change drastically with the size of the clusters. Because physical and chemical properties of clusters can be adjusted simply by changing the cluster's size, different applications of atomic clusters were proposed. One example is the catalytic activity of clusters of specific sizes in different chemical reactions. Another example is a potential application of atomic clusters in microelectronics, where their band gaps can be adjusted by simply changing cluster sizes. In recent years significant advances in experimental techniques allow one to synthesize and study atomic clusters of specified sizes. However, the interpretation of the results is often difficult. The theoretical methods are frequently used to help in interpretation of complex experimental data. Most of the theoretical approaches have been based on empirical or semiempirical methods. These methods allow one to study large and small dusters using the same approximations. However, since empirical and semiempirical methods rely on simple models with many parameters, it is often difficult to estimate the quantitative and even qualitative accuracy of the results. On the other hand, because of significant advances in quantum chemical methods and computer capabilities, it is now possible to do high quality ab-initio calculations not only on systems of few atoms but on clusters of practical interest as well. In addition to accurate results for specific clusters, such methods can be used for benchmarking of different empirical and semiempirical approaches. The atomic clusters studied in this work contain from a few atoms to

  5. Hybridization schemes for clusters

    NASA Astrophysics Data System (ADS)

    Wales, David J.

    The concept of an optimum hybridization scheme for cluster compounds is developed with particular reference to electron counting. The prediction of electron counts for clusters and the interpretation of the bonding is shown to depend critically upon the presumed hybridization pattern of the cluster vertex atoms. This fact has not been properly appreciated in previous work, particularly in applications of Stone's tensor surface harmonic (TSH) theory, but is found to be a useful tool when dealt with directly. A quantitative definition is suggested for the optimum cluster hybridization pattern based directly upon the ease of interpretation of the molecular orbitals, and results are given for a range of species. The relationship of this scheme to the detailed cluster geometry is described using Löwdin's partitioned perturbation theory, and the success and range of application of TSH theory are discussed.

  6. Scheffersomyces parashehatae f.a., sp. nov., Scheffersomyces xylosifermentans f.a., sp. nov., Candida broadrunensis sp. nov. and Candida manassasensis sp. nov., novel yeasts associated with wood-ingesting insects, and their ecological and biofuel implications.

    PubMed

    Suh, Sung-Oui; Houseknecht, Janice L; Gujjari, Pushpa; Zhou, Jianlong J

    2013-11-01

    During a survey of yeasts associated with wood-ingesting insects, 69 strains in the Scheffersomyces clade and related taxa were isolated from passalid and tenebrionid beetles and the decayed wood inhabited by them. The majority of these yeasts was found to be capable of fermenting xylose, and was recognized as Scheffersomyces stipitis or its close relative Scheffersomyces illinoinensis, which are known to be associated with wood-decaying beetles and rotten wood. Yeasts in 'Scheffersomyces' ( = Candida) ergatensis and 'Scheffersomyces' ( = Candida) coipomoensis were also frequently isolated. The remaining six strains were identified as representing four novel species in the genera Scheffersomyces and Candida based on multilocus sequence analyses of nuclear rRNA genes and four protein-coding genes, as well as other taxonomic characteristics. Two xylose-fermenting species, Scheffersomyces parashehatae f.a., sp. nov. (type strain ATCC MYA-4653(T) = CBS 12535(T) = EH045(T); MycoBank MB805440) and Scheffersomyces xylosifermentans f.a., sp. nov. (type strain ATCC MYA-4859(T) = CBS 12540(T) = MY10-052(T); MycoBank MB805441), formed a clade with Scheffersomyces shehatae and related Scheffersomyces species. Interestingly, S. xylosifermentans can survive at 40 °C, which is a rare property among xylose-fermenting yeasts. Candida broadrunensis sp. nov. (type strain ATCC MYA-4650(T) = CBS 11838(T) = EH019(T); MycoBank MB805442) is a sister taxon of C. ergatensis, while Candida manassasensis sp. nov. (type strain ATCC MYA-4652(T) = CBS 12534(T) = EH030(T); MycoBank MB805443) is closely related to Candida palmioleophila in the Candida glaebosa clade. The multilocus DNA sequence comparisons in this study suggest that the genus Scheffersomyces needs to be circumscribed to the species near S. stipitis (type species) and S. shehatae that can be characterized by the ability to ferment xylose. PMID:24014624

  7. Two yeast species Cystobasidium psychroaquaticum f.a. sp. nov. and Cystobasidium rietchieii f.a. sp. nov. isolated from natural environments, and the transfer of Rhodotorula minuta clade members to the genus Cystobasidium.

    PubMed

    Yurkov, A M; Kachalkin, A V; Daniel, H M; Groenewald, M; Libkind, D; de Garcia, V; Zalar, P; Gouliamova, D E; Boekhout, T; Begerow, D

    2015-01-01

    Many species of dimorphic basidiomycetes are known only in their asexual phase and typically those pigmented in different hues of red have been classified in the large polyphyletic genus Rhodotorula. These yeasts are ubiquitous and include a few species of some clinical relevance. The phylogenetic distribution of Rhodotorula spans three classes: Microbotryomycetes, Cystobasidiomycetes and Exobasidiomycetes. Here, the presented multi-gene analyses resolved phylogenetic relationships between the second largest group of Rhodotorula and the mycoparasite Cystobasidium fimetarium (Cystobasidiales, Cystobasidiomycetes, Pucciniomycotina). Based on the results, we propose the transfer of nine species belonging to the Rhodotorula minuta clade into the genus Cystobasidium. As a result, the clinically relevant species R. minuta will be renamed Cystobasidium minutum. This proposal follows ongoing reassessments of the anamorphic genus Rhodotorula reducing the polyphyly of this genus. The delimitation of the R. minuta clade from Rhodotorula species comprised in Sporidiobolales including the type species Rhodotorula glutinis is an important step to overcome obsolete generic placements of asexual basidiomycetous yeasts. Our proposal will also help to distinguish most common red yeasts from clinical samples such as members of Sporidiobolales and Cystobasidiales. The diagnosis of the genus Cystobasidium is amended by including additional characteristics known for the related group of species. The taxonomic change enables us to classify two novel species with the phylogenetically related members of the R. minuta clade in Cystobasidium. The recently from natural environments isolated species are described here as Cystobasidium psychroaquaticum f.a. sp. nov. (K-833(T) = KBP 3881(T) = VKPM Y-3653(T) = CBS 11769(T) = MUCL 52875(T) = DSM 27713(T)) and Cystobasidium rietchiei f.a. sp. nov. (K-780(T) = KBP 4220(T) = VKPM Y-3658(T) = CBS 12324(T) = MUCL 53589(T

  8. On the Relaxation of Superstring Axion Mini-Clusters

    NASA Astrophysics Data System (ADS)

    Pollock, M. D.

    The cosmological axion theory leads to the prediction of axionic mini-clusters of mass M 10-9Msolar, which form at the time te of equipartition of matter and radiation. By applying the two-body relaxation formula of Spitzer and Hart, we show, for the heterotic superstring theory of Gross et al., that these mini-clusters, considered as point masses, themselves cluster into axion mini-stars of mass M0≈ 10-2 (1 + λ 2)3/4 g3sMsun within the age of the Universe t0 only if they are located within a distance R 0.1 pc of the Galactic Center. Here, λ ≡ fB/fA is the ratio of the second to model-independent axion decay constants, assuming the QCD decay constant to be in the range 2.18 x 1016 g2s <= fa ≡ (1)/(2)(1+λ 2)1/2 fA/GeV< ˜ 2.4x 1018, and g2s ≈ 1.4 is the strong-interaction coupling parameter. Thus, if axion mini-stars are to explain the microlensing observations by the EROS and MACHO groups towards the Galactic Bulge and the Large and Small Magellanic Clouds, then a collisionless relaxation mechanism is required, as proposed by Seidel and Suen (essentially the violent relaxation of Lynden-Bell), or the four-axion self-interaction effect considered by Tkachev.

  9. Document clustering methods, document cluster label disambiguation methods, document clustering apparatuses, and articles of manufacture

    DOEpatents

    Sanfilippo, Antonio; Calapristi, Augustin J.; Crow, Vernon L.; Hetzler, Elizabeth G.; Turner, Alan E.

    2009-12-22

    Document clustering methods, document cluster label disambiguation methods, document clustering apparatuses, and articles of manufacture are described. In one aspect, a document clustering method includes providing a document set comprising a plurality of documents, providing a cluster comprising a subset of the documents of the document set, using a plurality of terms of the documents, providing a cluster label indicative of subject matter content of the documents of the cluster, wherein the cluster label comprises a plurality of word senses, and selecting one of the word senses of the cluster label.

  10. Deficiency of UBE2T, the E2 Ubiquitin Ligase Necessary for FANCD2 and FANCI Ubiquitination, Causes FA-T Subtype of Fanconi Anemia.

    PubMed

    Rickman, Kimberly A; Lach, Francis P; Abhyankar, Avinash; Donovan, Frank X; Sanborn, Erica M; Kennedy, Jennifer A; Sougnez, Carrie; Gabriel, Stacey B; Elemento, Olivier; Chandrasekharappa, Settara C; Schindler, Detlev; Auerbach, Arleen D; Smogorzewska, Agata

    2015-07-01

    Fanconi anemia (FA) is a rare bone marrow failure and cancer predisposition syndrome resulting from pathogenic mutations in genes encoding proteins participating in the repair of DNA interstrand crosslinks (ICLs). Mutations in 17 genes (FANCA-FANCS) have been identified in FA patients, defining 17 complementation groups. Here, we describe an individual presenting with typical FA features who is deficient for the ubiquitin-conjugating enzyme (E2), UBE2T. UBE2T is known to interact with FANCL, the E3 ubiquitin-ligase component of the multiprotein FA core complex, and is necessary for the monoubiquitination of FANCD2 and FANCI. Proband fibroblasts do not display FANCD2 and FANCI monoubiquitination, do not form FANCD2 foci following treatment with mitomycin C, and are hypersensitive to crosslinking agents. These cellular defects are complemented by expression of wild-type UBE2T, demonstrating that deficiency of the protein UBE2T can lead to Fanconi anemia. UBE2T gene gains an alias of FANCT. PMID:26119737

  11. A NIMA-related kinase, Fa2p, localizes to a novel site in the proximal cilia of Chlamydomonas and mouse kidney cells.

    PubMed

    Mahjoub, Moe R; Qasim Rasi, M; Quarmby, Lynne M

    2004-11-01

    Polycystic kidney disease and related syndromes involve dysregulation of cell proliferation in conjunction with ciliary defects. The relationship between cilia and cell cycle is enigmatic, but it may involve regulation by the NIMA-family of kinases (Neks). We previously showed that the Nek Fa2p is important for ciliary function and cell cycle in Chlamydomonas. We now show that Fa2p localizes to an important regulatory site at the proximal end of cilia in both Chlamydomonas and a mouse kidney cell line. Fa2p also is associated with the proximal end of centrioles. Its localization is dynamic during the cell cycle, following a similar pattern in both cell types. The cell cycle function of Fa2p is kinase independent, whereas its ciliary function is kinase dependent. Mice with mutations in Nek1 or Nek8 have cystic kidneys; therefore, our discovery that a member of this phylogenetic group of Nek proteins is localized to the same sites in Chlamydomonas and kidney epithelial cells suggests that Neks play conserved roles in the coordination of cilia and cell cycle progression. PMID:15371535

  12. Use of conventional surfactant media as surrogates for FaSSIF in simulating in vivo dissolution of BCS class II drugs.

    PubMed

    Lehto, Paula; Kortejärvi, Hanna; Liimatainen, Anni; Ojala, Krista; Kangas, Heli; Hirvonen, Jouni; Tanninen, Veli Pekka; Peltonen, Leena

    2011-08-01

    The usefulness of selected conventional surfactant media to enhance dissolution of BCS class II drugs similarly to fasted state simulated intestinal fluid (FaSSIF) and to predict the absorption of drugs in vivo was evaluated. Dissolution behavior of danazol (Danol), spironolactone (Spiridon) and N74 (phase I compound) was compared between FaSSIF, containing physiological levels of sodium taurocholate (STC) and lecithin, and dissolution media containing various concentrations of anionic surfactant, sodium lauryl sulfate (SLS) or non-ionic surfactant, polysorbate (Tween) 80. Although these media differed largely in their solubilization ability, micelle size, diffusivity and surface tension, similar dissolution enhancing levels were achieved between FaSSIF and drug-specific concentrations of conventional surfactants. The dissolution enhancement was shown, however, to be important only for danazol and N74, molecules that are characterized by high hydrophobicity. An in vivo pharmacokinetic dog study was carried out with N74. Comparison of observed plasma profiles with simulated profiles obtained using compartmental absorption and transit model (CAT) indicated that 0.1% SLS medium was the best to predict in vivo plasma profiles and pharmacokinetic parameters (C(max) and AUC). This study demonstrates the potential of substituting FaSSIF with more simple and cost-effective conventional surfactant media. Use of in vivo prognostic amounts of synthetic surfactants in dissolution testing could largely assist in industrial drug development as well as in quality control purposes. PMID:21329757

  13. Dissolution Profile of Mefenamic Acid Solid Dosage Forms in Two Compendial and Biorelevant (FaSSIF) Media.

    PubMed

    Nurhikmah, Wilda; Sumirtapura, Yeyet Cahyati; Pamudji, Jessie Sofia

    2016-01-01

    Mefenamic acid is a non-steroidal anti-inflammatory drug (NSAID) that is widely used for the treatment of mild-to-moderate pain. Mefenamic acid belongs to the Biopharmaceutical Classification System (BCS) class II drug which has lower water solubility but high permeability. There are two different compendial methods available for dissolution tests of mefenamic acid solid dosage forms, i.e. methods of United States Pharmacopeia 37 (USP) and Pharmacopoeia of the People's Republic of China 2010 (PPRC). Indonesian Pharmacopeia V ed. (FI) adopted the USP method. On the other hand, many researches focused on the use of a 'biorelevant' medium to develop the dissolution test method. The aim of this research was to study the dissolution profile of mefenamic acid from its solid dosage forms (caplet and capsule) available in the Indonesian market with three different dissolution medium: USP, PPRC, and biorelevant fasted simulated small intestinal fluid (FaSSIF) media. The tested products consisted of the innovator's product (available only in caplet dosage form, FN caplet) and generic products (available as caplet and capsule). The dissolution test of the drug products in all dissolution media was performed in 900 mL of medium using apparatus II (paddle) at a temperature of 37°C and rotation speed of 75 rpm, except for the capsule product and for USP medium, both of which tests were done using apparatus I (basket) with rotation speed of 100 rpm. The solubility test of mefenamic acid was carried out in all media at temperature of 37°C. The result obtained from the solubility test showed that the the highest solubility of mefenamic acid was obtained in USP medium (approximately 2 mg/mL), followed by PPRC medium (about 0.5 mg/mL), and FaSSIF medium (approximately 0.06 mg/ml). In the dissolution test, percentage of drug dissolved in in the USP and PPRC media after 45 min for all products reached more than 75%, except for the PN caplet in USP medium which reached only about 44

  14. Dissolution Profile of Mefenamic Acid Solid Dosage Forms in Two Compendial and Biorelevant (FaSSIF) Media

    PubMed Central

    Nurhikmah, Wilda; Sumirtapura, Yeyet Cahyati; Pamudji, Jessie Sofia

    2016-01-01

    Mefenamic acid is a non-steroidal anti-inflammatory drug (NSAID) that is widely used for the treatment of mild-to-moderate pain. Mefenamic acid belongs to the Biopharmaceutical Classification System (BCS) class II drug which has lower water solubility but high permeability. There are two different compendial methods available for dissolution tests of mefenamic acid solid dosage forms, i.e. methods of United States Pharmacopeia 37 (USP) and Pharmacopoeia of the People’s Republic of China 2010 (PPRC). Indonesian Pharmacopeia V ed. (FI) adopted the USP method. On the other hand, many researches focused on the use of a ‘biorelevant’ medium to develop the dissolution test method. The aim of this research was to study the dissolution profile of mefenamic acid from its solid dosage forms (caplet and capsule) available in the Indonesian market with three different dissolution medium: USP, PPRC, and biorelevant fasted simulated small intestinal fluid (FaSSIF) media. The tested products consisted of the innovator’s product (available only in caplet dosage form, FN caplet) and generic products (available as caplet and capsule). The dissolution test of the drug products in all dissolution media was performed in 900 mL of medium using apparatus II (paddle) at a temperature of 37°C and rotation speed of 75 rpm, except for the capsule product and for USP medium, both of which tests were done using apparatus I (basket) with rotation speed of 100 rpm. The solubility test of mefenamic acid was carried out in all media at temperature of 37°C. The result obtained from the solubility test showed that the the highest solubility of mefenamic acid was obtained in USP medium (approximately 2 mg/mL), followed by PPRC medium (about 0.5 mg/mL), and FaSSIF medium (approximately 0.06 mg/ml). In the dissolution test, percentage of drug dissolved in in the USP and PPRC media after 45 min for all products reached more than 75%, except for the PN caplet in USP medium which reached only

  15. Analysis of F/A-18 Tail Buffet Data Acquired in the 80- by 120-Foot Wind Tunnel

    NASA Technical Reports Server (NTRS)

    James, Kevin D.; Meyn, Larry A.; Schmitz, Fredric H. (Technical Monitor)

    1994-01-01

    Tail buffet studies were conducted on a full-scale, production, F/A-18 fighter aircraft in the 80- by 120-Foot Wind Tunnel of the National Full-Scale Aerodynamic Complex at NASA Ames Research Center at Moffett Field, California. Tail buffet data were acquired over an angle-of-attack range of +20 deg to +40 deg, a side-slip range of -16 deg to + 16 deg, and at wind speeds up to 100 knots. The maximum speed corresponds to a Reynolds number of l2.3 x l0(exp 6) based on mean aerodynamic chord and a Mach number of 0. 15. The port, vertical tail fin was instrumented with ninety-six surface-pressure transducers, arranged in six by eight arrays, on each side of the fin. ne aircraft was also equipped with a removable Leading-Edge Extension (LEX) fence whose purpose is to reduce tail-buffet loads. Current analysis methods for the unsteady aerodynamic pressures and loads are described. Only results for the zero side-slip condition are to be presented, both with and without the LEX fence. Results of the time-averaged, power-spectral analysis are presented for the tail fin bending moments which are derived from the integrated pressure field. Local wave velocities on the tail surfaces are calculated from pressure correlations. It was found that the LEX fence significantly reduces the magnitude of the root-mean-square pressures and bending moments. Scaling and repeatability issues are addressed by comparing the present full scale results for pressures at the 60%-span and 45%-chord location with previous full-scale F/A-18 tail-buffet test in the 80- by 120- Foot Wind Tunnel, and with several small-scale tests. The comparisons show that the tail buffet frequency scales very well with tail chord and free-stream velocity, and that there is good agreement with the previous full-scale test. Root-mean-square pressures and power spectra do not scale as well as the frequency results. Addition of a LEX fence caused tail-buffet loads to be reduced at all model scales.

  16. Structure of the GcpE (IspG)-MEcPP complex from Thermus thermophilus.

    PubMed

    Rekittke, Ingo; Jomaa, Hassan; Ermler, Ulrich

    2012-09-21

    Isoprenoid precursor biosynthesis occurs through the mevalonate or the methylerythritol phosphate (MEP) pathway, used i.e., by humans and by many human pathogens, respectively. In the MEP pathway, 2-C-methyl-D-erythritol-2,4-cyclo-diphosphate (MEcPP) is converted to (E)-1-hydroxy-2-methyl-but-2-enyl-4-diphosphate (HMBPP) by the iron-sulfur cluster enzyme HMBPP synthase (GcpE). The presented X-ray structure of the GcpE-MEcPP complex from Thermus thermophilus at 1.55Å resolution provides valuable information about the catalytic mechanism and for rational inhibitor design. MEcPP binding inside the TIM-barrel funnel induces a 60° rotation of the [4Fe-4S] cluster containing domain onto the TIM-barrel entrance. The apical iron of the [4Fe-4S] cluster ligates with the C3 oxygen atom of MEcPP. PMID:22967895

  17. Prismatic louver active façades for natural illumination and thermal energy gain in high-rise and commercial buildings

    NASA Astrophysics Data System (ADS)

    Vlachokostas, A.; Volkmann, C.; Madamopoulos, N.

    2013-06-01

    High-rise and commercial buildings in urban centers present a great challenge in terms of their energy consumption. Due to maximization of rentable square footage, the preferred urban façade system over the past 50 years has been the "curtain wall", only a few inches thick and comprised of modular steel or aluminum framing and predominant glass infills. The perceived Achilles heel of these modern glass façade systems is their thermal inefficiency: They are inadequate thermal barriers and exhibit excessive solar gain. The excessive solar gain has a negative impact on lighting and cooling loads of the entire building. This negative impact will be further exacerbated with rising energy costs. However, rather than view the glass façade's uncontrolled solar gain merely as a weakness contributing to higher energy consumption, the condition could indeed be considered as related to an energy solution. These glass façades can be retrofitted to operate as a provider of daylight and energy for the rest of the building, taking advantage of the overexposure to the sun. With today's technology, the sun's abundant renewable energy can be the driving force for the energy transition of these building envelopes. Illumination, thermal energy, and electricity production can be directly supplied from the sun, and when correctly and efficiently managed, they can lead to a significantly less energy-intensive building stock. We propose a multi-purpose, prismatic, louver-based façade to perform both daylight and thermal energy harvesting with a goal of offering a better daylight environment for the occupants, and reduce the energy consumption and carbon footprint of the building. While decentralized air-conditioning units are commonly accepted as façade "plug-ins", such decentralization could be utilized with more benefits by passively managing the interior space conditions, without using any extra power. Just as living organisms respond and adapt to the environmental changes in

  18. Statistical properties of convex clustering

    PubMed Central

    Tan, Kean Ming; Witten, Daniela

    2016-01-01

    In this manuscript, we study the statistical properties of convex clustering. We establish that convex clustering is closely related to single linkage hierarchical clustering and k-means clustering. In addition, we derive the range of the tuning parameter for convex clustering that yields a non-trivial solution. We also provide an unbiased estimator of the degrees of freedom, and provide a finite sample bound for the prediction error for convex clustering. We compare convex clustering to some traditional clustering methods in simulation studies.

  19. Partially supervised speaker clustering.

    PubMed

    Tang, Hao; Chu, Stephen Mingyu; Hasegawa-Johnson, Mark; Huang, Thomas S

    2012-05-01

    Content-based multimedia indexing, retrieval, and processing as well as multimedia databases demand the structuring of the media content (image, audio, video, text, etc.), one significant goal being to associate the identity of the content to the individual segments of the signals. In this paper, we specifically address the problem of speaker clustering, the task of assigning every speech utterance in an audio stream to its speaker. We offer a complete treatment to the idea of partially supervised speaker clustering, which refers to the use of our prior knowledge of speakers in general to assist the unsupervised speaker clustering process. By means of an independent training data set, we encode the prior knowledge at the various stages of the speaker clustering pipeline via 1) learning a speaker-discriminative acoustic feature transformation, 2) learning a universal speaker prior model, and 3) learning a discriminative speaker subspace, or equivalently, a speaker-discriminative distance metric. We study the directional scattering property of the Gaussian mixture model (GMM) mean supervector representation of utterances in the high-dimensional space, and advocate exploiting this property by using the cosine distance metric instead of the euclidean distance metric for speaker clustering in the GMM mean supervector space. We propose to perform discriminant analysis based on the cosine distance metric, which leads to a novel distance metric learning algorithm—linear spherical discriminant analysis (LSDA). We show that the proposed LSDA formulation can be systematically solved within the elegant graph embedding general dimensionality reduction framework. Our speaker clustering experiments on the GALE database clearly indicate that 1) our speaker clustering methods based on the GMM mean supervector representation and vector-based distance metrics outperform traditional speaker clustering methods based on the “bag of acoustic features” representation and statistical

  20. Dwarfs in Coma Cluster

    NASA Technical Reports Server (NTRS)

    2007-01-01

    [figure removed for brevity, see original site] Click on image for larger poster version

    This false-color mosaic of the central region of the Coma cluster combines infrared and visible-light images to reveal thousands of faint objects (green). Follow-up observations showed that many of these objects, which appear here as faint green smudges, are dwarf galaxies belonging to the cluster. Two large elliptical galaxies, NGC 4889 and NGC 4874, dominate the cluster's center. The mosaic combines visible-light data from the Sloan Digital Sky Survey (color coded blue) with long- and short-wavelength infrared views (red and green, respectively) from NASA's Spitzer Space Telescope.

  1. H-cluster stars

    NASA Astrophysics Data System (ADS)

    Lai, X. Y.; Gao, C. Y.; Xu, R. X.

    2013-06-01

    The study of dense matter at ultrahigh density has a very long history, which is meaningful for us to understand not only cosmic events in extreme circumstances but also fundamental laws of physics. It is well known that the state of cold matter at supranuclear density depends on the non-perturbative nature of quantum chromodynamics (QCD) and is essential for modelling pulsars. A so-called H-cluster matter is proposed in this paper as the nature of dense matter in reality. In compact stars at only a few nuclear densities but low temperature, quarks could be interacting strongly with each other there. That might render quarks grouped in clusters, although the hypothetical quark clusters in cold dense matter have not been confirmed due to the lack of both theoretical and experimental evidence. Motivated by recent lattice QCD simulations of the H-dibaryons (with structure uuddss), we therefore consider here a possible kind of quark clusters, H-clusters, that could emerge inside compact stars during their initial cooling as the dominant components inside (the degree of freedom could then be H-clusters there). Taking into account the in-medium stiffening effect, we find that at baryon densities of compact stars H-cluster matter could be more stable than nuclear matter. We also find that for the H-cluster matter with lattice structure, the equation of state could be so stiff that it would seem to be `superluminal' in the most dense region. However, the real sound speed for H-cluster matter is in fact difficult to calculate, so at this stage we do not put constraints on our model from the usual requirement of causality. We study the stars composed of H-clusters, i.e. H-cluster stars, and derive the dependence of their maximum mass on the in-medium stiffening effect, showing that the maximum mass could be well above 2 M⊙ as observed and that the resultant mass-radius relation fits the measurement of the rapid burster under reasonable parameters. Besides a general

  2. Extending Beowulf Clusters

    USGS Publications Warehouse

    Steinwand, Daniel R.; Maddox, Brian; Beckmann, Tim; Hamer, George

    2003-01-01

    Beowulf clusters can provide a cost-effective way to compute numerical models and process large amounts of remote sensing image data. Usually a Beowulf cluster is designed to accomplish a specific set of processing goals, and processing is very efficient when the problem remains inside the constraints of the original design. There are cases, however, when one might wish to compute a problem that is beyond the capacity of the local Beowulf system. In these cases, spreading the problem to multiple clusters or to other machines on the network may provide a cost-effective solution.

  3. In Situ Confinement of Ultrasmall Pd Clusters within Nanosized Silicalite-1 Zeolite for Highly Efficient Catalysis of Hydrogen Generation.

    PubMed

    Wang, Ning; Sun, Qiming; Bai, Risheng; Li, Xu; Guo, Guanqi; Yu, Jihong

    2016-06-22

    Well-dispersed and ultrasmall Pd clusters in nanosized silicalite-1 (MFI) zeolite have been prepared under direct hydrothermal conditions using [Pd(NH2CH2CH2NH2)2]Cl2 as precursor. High-resolution scanning transmission electron microscopy studies indicate that the Pd clusters are encapsulated within the intersectional channels of MFI, and the Pd clusters in adjacent channels visually aggregate, forming nanoparticles (NPs) of ∼1.8 nm. The resultant catalysts show an excellent activity and highly efficient H2 generation toward the complete decomposition of formic acid (FA) under mild conditions. Notably, thanks to the further reduced Pd NP size (∼1.5 nm) and the additionally introduced basic sites, the Pd/S-1-in-K catalyst affords turnover frequency values up to 856 h(-1) at 25 °C and 3027 h(-1) at 50 °C. The easy in situ confinement synthesis of metal clusters in zeolites endows the catalysts with superior catalytic activities, excellent recyclability, and high thermal stability, thus opening new perspectives for the practical application of FA as a viable and effective H2 storage material for use in fuel cells. PMID:27248462

  4. Implantation and post-annealing characteristics when impinging small B n clusters into silicon at low fluence

    NASA Astrophysics Data System (ADS)

    Liang, J. H.; Han, H. M.

    2005-01-01

    This study investigated the similarities and differences between B1 monomer and Bn cluster ion implantation into silicon. Small polyatomic boron ions ( Bn- , n = 1-4) with the same atomic boron kinetic energy (20 keV/atom) and atomic fluence (5 × 1013 atoms/cm2) were used. In the simulation, the widely-used SRIM computer code was employed to calculate the as-implanted boron and damage depth profiles of B1 monomer ion implantation in order to make comparisons with experimental results. In the experimental one, the B1 monomer and Bn cluster ions extracted from a tandem accelerator were used to perform ion implantation. Post-annealing methods included one-step (RTA) and two-step (FA + RTA) treatments, where RTA denoted high-temperature rapid thermal annealing at 1050 °C for 10 s and FA represented low-temperature furnace annealing at 550 °C for 1 h. The results revealed that all four as-implanted range parameters (average range, longitudinal range straggling, skewness, kurtosis) increase and tend to saturate as the cluster size increases when compared to those of SRIM-calculated results for the B1 implant. Furthermore, the peculiar damage structures produced by different Bn cluster ions lead to various behaviors in both diffusing and activating boron atoms.

  5. Combining cluster number counts and galaxy clustering

    NASA Astrophysics Data System (ADS)

    Lacasa, Fabien; Rosenfeld, Rogerio

    2016-08-01

    The abundance of clusters and the clustering of galaxies are two of the important cosmological probes for current and future large scale surveys of galaxies, such as the Dark Energy Survey. In order to combine them one has to account for the fact that they are not independent quantities, since they probe the same density field. It is important to develop a good understanding of their correlation in order to extract parameter constraints. We present a detailed modelling of the joint covariance matrix between cluster number counts and the galaxy angular power spectrum. We employ the framework of the halo model complemented by a Halo Occupation Distribution model (HOD). We demonstrate the importance of accounting for non-Gaussianity to produce accurate covariance predictions. Indeed, we show that the non-Gaussian covariance becomes dominant at small scales, low redshifts or high cluster masses. We discuss in particular the case of the super-sample covariance (SSC), including the effects of galaxy shot-noise, halo second order bias and non-local bias. We demonstrate that the SSC obeys mathematical inequalities and positivity. Using the joint covariance matrix and a Fisher matrix methodology, we examine the prospects of combining these two probes to constrain cosmological and HOD parameters. We find that the combination indeed results in noticeably better constraints, with improvements of order 20% on cosmological parameters compared to the best single probe, and even greater improvement on HOD parameters, with reduction of error bars by a factor 1.4-4.8. This happens in particular because the cross-covariance introduces a synergy between the probes on small scales. We conclude that accounting for non-Gaussian effects is required for the joint analysis of these observables in galaxy surveys.

  6. Mantis BT Cluster Support

    2009-06-05

    The software is a modidication to the Mantis BT V1.5 open source application provided by the mantis BT group to support cluster web servers. It also provides various cosmetic modifications used a LLNL.

  7. Cyclostomes Lack Clustered Protocadherins.

    PubMed

    Ravi, Vydianathan; Yu, Wei-Ping; Pillai, Nisha E; Lian, Michelle M; Tay, Boon-Hui; Tohari, Sumanty; Brenner, Sydney; Venkatesh, Byrappa

    2016-02-01

    The brain, comprising billions of neurons and intricate neural networks, is arguably the most complex organ in vertebrates. The diversity of individual neurons is fundamental to the neuronal network complexity and the overall function of the vertebrate brain. In jawed vertebrates, clustered protocadherins provide the molecular basis for this neuronal diversity, through stochastic and combinatorial expression of their various isoforms in individual neurons. Based on analyses of transcriptomes from the Japanese lamprey brain and sea lamprey embryos, genome assemblies of the two lampreys, and brain expressed sequence tags of the inshore hagfish, we show that extant jawless vertebrates (cyclostomes) lack the clustered protocadherins. Our findings indicate that the clustered protocadherins originated from a nonclustered protocadherin in the jawed vertebrate ancestor, after the two rounds of whole-genome duplication. In the absence of clustered protocadherins, cyclostomes might have evolved novel molecules or mechanisms for generating neuronal diversity which remains to be discovered. PMID:26545918

  8. Dynamic Bayesian clustering.

    PubMed

    Fowler, Anna; Menon, Vilas; Heard, Nicholas A

    2013-10-01

    Clusters of time series data may change location and memberships over time; in gene expression data, this occurs as groups of genes or samples respond differently to stimuli or experimental conditions at different times. In order to uncover this underlying temporal structure, we consider dynamic clusters with time-dependent parameters which split and merge over time, enabling cluster memberships to change. These interesting time-dependent structures are useful in understanding the development of organisms or complex organs, and could not be identified using traditional clustering methods. In cell cycle data, these time-dependent structure may provide links between genes and stages of the cell cycle, whilst in developmental data sets they may highlight key developmental transitions. PMID:24131050

  9. A xylanase from Streptomyces sp. FA1: heterologous expression, characterization, and its application in Chinese steamed bread.

    PubMed

    Xu, Yang; Wu, Jing; Zheng, Kaixuan; Wu, Dan

    2016-05-01

    Xylanases (EC 3.2.1.8) are hydrolytic enzymes that have found widespread application in the food, feed, and paper-pulp industries. Streptomyces sp. FA1 xynA was expressed as a secreted protein in Pichia pastoris, and the xylanase was applied to the production of Chinese steamed bread for the first time. The optimal pH and the optimal temperature of XynA were 5.5 and 60 °C, respectively. Using beechwood as substrate, the K m and V max were 2.408 mg mL(-1) and 299.3 µmol min(-1) mg(-1), respectively. Under optimal conditions, a 3.6-L bioreactor produced 1374 U mL(-1) of XynA activity at a protein concentration of 6.3 g L(-1) after 132 h of fermentation. Use of recombinant XynA led to a greater increase in the specific volume of the CSB than could be achieved using commercial xylanase under optimal conditions. This study provides the basis for the application of the enzyme in the baking industry. PMID:26803505

  10. Measurement of enhanced heat transfer coefficient with perforated twisted tape inserts during condensation of R-245fa

    NASA Astrophysics Data System (ADS)

    Khatua, A. K.; Kumar, P.; Singh, H. N.; Kumar, R.

    2016-04-01

    The experimental conductive heat transfer results for flow through inserted perforated twisted tapes in a horizontal tube during condensation of pure R-245fa vapor. The test section consisting of two separate coaxial double pipes assembled in series, acted like a counter flow heat exchanger, where the refrigerant condensed inside the inner tube by rejecting heat to the cooling water flowing inside the outer tube in reversed direction. Data for three perforated twisted tapes having constant twist ratio of 7.1 mm and pitch of perforation as 12.5, 25.0 and 37.5 mm, inserted one by one in full length of test condenser by varying refrigerant mass flux from 100 to 200 kg/m2 s in steps of 50 kg/m2 s for the range of vapor quality from 0.1 to 0.9, were collected together with flow and without insert (plain tube). It has been found that the perforated twisted tape insert having pitch of perforation equal to in order of 12.5 mm gives the highest value of average heat transfer coefficient and is of the order of 37.5 % more than that of the plain one and the correlation predicts the experimental data within an error band of ±15 %.

  11. In-Flight Wing Pressure Distributions for the NASA F/A-18A High Alpha Research Vehicle

    NASA Technical Reports Server (NTRS)

    Davis, Mark C.; Saltzman, John A.

    2000-01-01

    Pressure distributions on the wings of the F/A-18A High Alpha Research Vehicle (HARV) were obtained using both flush-mounted pressure orifices and surface-mounted pressure tubing. During quasi-stabilized 1-g flight, data were gathered at ranges for angle of attack from 5 deg to 70 deg, for angle of sideslip from -12 deg to +12 deg, and for Mach from 0.23 to 0.64, at various engine settings, and with and without the leading edge extension fence installed. Angle of attack strongly influenced the wing pressure distribution, as demonstrated by a distinct flow separation pattern that occurred between the range from 15 deg to 30 deg. Influence by the leading edge extension fence was evident on the inboard wing pressure distribution, but little influence was seen on the outboard portion of the wing. Angle-of-sideslip influence on wing pressure distribution was strongest at low angle of attack. Influence of Mach number was observed in the regions of local supersonic flow, diminishing as angle of attack was increased. Engine throttle setting had little influence on the wing pressure distribution.

  12. The Financial Management Environment (FaME): A prototype interactive hypertext-based financial planning and reporting system

    SciTech Connect

    Barnes, K.D.; Donato, J.M.; Flanagan, D.M.

    1995-11-01

    It is critical in every government, research, and industrial organization that accurate and timely financial information be made available at all levels so that, project and business decisions can be made within funding constraints. The FaME prototype implemented at Oak Ridge National Laboratory extracts financial data from a legacy system, builds easy-to-understand reports and graphs, and presents them on-line so that people at all levels in an organization can assess the financial status of individual projects or entire organizations. Reports are presented in hypertext and graphical formats that can be read with popular World Wide Web browsers such as NCSA Mosaic or Netscape. All reports are hyper-linked in a natural way to simplify navigation and information retrieval. To protect potentially sensitive information, FAME provides access control so that individuals can retrieve only the information that is required for them to carry out their financial duties. In addition to the reports and graphs, FAME includes budget building tools to provide for financial planning. Another primary feature is that the prototype 1 utilizes equipment that already exists on the user`s desktop. The overall goal of the FAME system is to provide users with precise and meaningful information on the financial status of an organization or project at a glance.

  13. Flow Visualization Techniques in Wind Tunnel Tests of a Full-Scale F/A-18 Aircraft

    NASA Technical Reports Server (NTRS)

    Lanser, Wendy R.; Botha, Gavin J.; James, Kevin D.; Bennett, Mark; Crowder, James P.; Cooper, Don; Olson, Lawrence (Technical Monitor)

    1994-01-01

    The proposed paper presents flow visualization performed during experiments conducted on a full-scale F/A-18 aircraft in the 80- by 120-Foot Wind-Tunnel at NASA Ames Research Center. The purpose of the flow-visualization experiments was to document the forebody and leading edge extension (LEX) vortex interaction along with the wing flow patterns at high angles of attack and low speed high Reynolds number conditions. This investigation used surface pressures in addition to both surface and off-surface flow visualization techniques to examine the flow field on the forebody, canopy, LEXS, and wings. The various techniques used to visualize the flow field were fluorescent tufts, flow cones treated with reflective material, smoke in combination with a laser light sheet, and a video imaging system for three-dimension vortex tracking. The flow visualization experiments were conducted over an angle of attack range from 20 deg to 45 deg and over a sideslip range from -10 deg to 10 deg. The various visualization techniques as well as the pressure distributions were used to understand the flow field structure. The results show regions of attached and separated flow on the forebody, canopy, and wings as well as the vortical flow over the leading-edge extensions. This paper will also present flow visualization comparisons with the F-18 HARV flight vehicle and small-scale oil flows on the F-18.

  14. A static investigation of the thrust vectoring system of the F/A-18 high-alpha research vehicle

    NASA Technical Reports Server (NTRS)

    Mason, Mary L.; Capone, Francis J.; Asbury, Scott C.

    1992-01-01

    A static (wind-off) test was conducted in the static test facility of the Langley 16-foot Transonic Tunnel to evaluate the vectoring capability and isolated nozzle performance of the proposed thrust vectoring system of the F/A-18 high alpha research vehicle (HARV). The thrust vectoring system consisted of three asymmetrically spaced vanes installed externally on a single test nozzle. Two nozzle configurations were tested: A maximum afterburner-power nozzle and a military-power nozzle. Vane size and vane actuation geometry were investigated, and an extensive matrix of vane deflection angles was tested. The nozzle pressure ratios ranged from two to six. The results indicate that the three vane system can successfully generate multiaxis (pitch and yaw) thrust vectoring. However, large resultant vector angles incurred large thrust losses. Resultant vector angles were always lower than the vane deflection angles. The maximum thrust vectoring angles achieved for the military-power nozzle were larger than the angles achieved for the maximum afterburner-power nozzle.

  15. High-Alpha Handling Qualities Flight Research on the NASA F/A-18 High Alpha Research Vehicle

    NASA Technical Reports Server (NTRS)

    Wichman, Keith D.; Pahle, Joseph W.; Bahm, Catherine; Davidson, John B.; Bacon, Barton J.; Murphy, Patrick C.; Ostroff, Aaron J.; Hoffler, Keith D.

    1996-01-01

    A flight research study of high-angle-of-attack handling qualities has been conducted at the NASA Dryden Flight Research Center using the F/A-18 High Alpha Research Vehicle (HARV). The objectives were to create a high-angle-of-attack handling qualities flight database, develop appropriate research evaluation maneuvers, and evaluate high-angle-of-attack handling qualities guidelines and criteria. Using linear and nonlinear simulations and flight research data, the predictions from each criterion were compared with the pilot ratings and comments. Proposed high-angle-of-attack nonlinear design guidelines and proposed handling qualities criteria and guidelines developed using piloted simulation were considered. Recently formulated time-domain Neal-Smith guidelines were also considered for application to high-angle-of-attack maneuvering. Conventional envelope criteria were evaluated for possible extension to the high-angle-of-attack regime. Additionally, the maneuvers were studied as potential evaluation techniques, including a limited validation of the proposed standard evaluation maneuver set. This paper gives an overview of these research objectives through examples and summarizes result highlights. The maneuver development is described briefly, the criteria evaluation is emphasized with example results given, and a brief discussion of the database form and content is presented.

  16. An Overview of Controls and Flying Qualities Technology on the F/A-18 High Alpha Research Vehicle

    NASA Technical Reports Server (NTRS)

    Pahle, Joseph W.; Wichman, Keith D.; Foster, John V.; Bundick, W. Thomas

    1996-01-01

    The NASA F/A-18 High Alpha Research Vehicle (HARV) has been the flight test bed of a focused technology effort to significantly increase maneuvering capability at high angles of attack. Development and flight test of control law design methodologies, handling qualities metrics, performance guidelines, and flight evaluation maneuvers are described. The HARV has been modified to include two research control effectors, thrust vectoring, and actuated forebody strakes in order to provide increased control power at high angles of attack. A research flight control system has been used to provide a flexible, easily modified capability for high-angle-of-attack research controls. Different control law design techniques have been implemented and flight-tested, including eigenstructure assignment, variable gain output feedback, pseudo controls, and model-following. Extensive piloted simulation has been used to develop nonlinear performance guide-lines and handling qualities criteria for high angles of attack. This paper reviews the development and evaluation of technologies useful for high-angle-of-attack control. Design, development, and flight test of the research flight control system, control laws, flying qualities specifications, and flight test maneuvers are described. Flight test results are used to illustrate some of the lessons learned during flight test and handling qualities evaluations.

  17. Molecular Phylogeny and Taxonomy of Yamadazyma dushanensis f.a., sp. nov., a Cellobiose-Fermenting Yeast Species from China.

    PubMed

    Wang, Yun; Ren, Yong-Cheng; Li, Ying; Hui, Feng-Li

    2015-08-01

    Three yeast strains of Yamadazyma dushanensis f.a., sp. nov. were isolated from rotten wood samples collected in the Dushan Forest Park, Nanyang, Henan Province, China. Sequence analyses of the D1/D2 domains of the large subunit rRNA gene and the internal transcribed spacer (ITS) regions revealed that this new species is located in the Yamadazyma clade (Debaryomycetaceae and Saccharomycetales), with three closely related species, namely, Yamadazyma terventina, Yamadazyma mexicana and Candida trypodendroni. The novel species differed from these three described species by 5-6 nt substitutions in the D1/D2 sequences. However, the ITS sequences of the new species were quite divergent from those of Y. terventina, Y. mexicana and C. trypodendroni with 12-18 nt substitutions. This new yeast species could assimilate cellobiose and other compounds related to rotting wood. The fermentation of cellobiose in Durham tubes was observed for the strains of this new yeast. The new species could also be distinguished from its closely related species, Y. terventina, Y. mexicana and C. trypodendroni, based on a number of morphological and physiological characteristics. The type strain is Y. dushanensis sp. nov. NYNU 14668 (T) (=CICC 33051(T) = CBS 13914(T)). PMID:26037380

  18. Wickerhamiella siamensis f.a., sp. nov., an endophytic and epiphytic yeast species isolated from sugar cane leaf.

    PubMed

    Khunnamwong, Pannida; Surussawadee, Janjira; Jindamorakot, Sasitorn; Limtong, Savitree

    2014-11-01

    Six strains representing a novel yeast species were isolated from tissue (DMKU-SE106(T), DMKU-SE110, DMKU-SE112 and DMKU-SE132) and the external surface (DMKU-SP335 and DMKU-SP406) of sugar cane leaves collected in Thailand. On the basis of morphological, biochemical, physiological and chemotaxonomic characteristics, and sequence analysis of the D1/D2 region of the LSU rRNA gene and the internal transcribed spacer (ITS) region, the six strains were found to represent a single novel species of the genus Wickerhamiella although the formation of ascospores was not observed. The sequences of the D1/D2 region of the LSU rRNA gene and ITS region of the six strains differed from each other by 0-2 and 2-3 nt substitutions, respectively. The novel species was related most closely to Candida infanticola but with 4.5-4.6% nucleotide substitutions in the D1/D2 region of the LSU rRNA gene and 6.6-7.1% nucleotide substitutions in the ITS region. The name Wickerhamiella siamensis f.a., sp. nov. is proposed. The type strain is DMKU-SE106(T) ( =BCC 61185(T) =NBRC 109697(T) =CBS 13331(T)). PMID:25168613

  19. Fully automated measurement of field-dependent AMS using MFK1-FA Kappabridge equipped with 3D rotator

    NASA Astrophysics Data System (ADS)

    Chadima, Martin; Studynka, Jan

    2013-04-01

    Low-field magnetic susceptibility of paramagnetic and diamagnetic minerals is field-independent by definition being also field-independent in pure magnetite. On the other hand, in pyrrhotite, hematite and high-Ti titanomagnetite it may be clearly field-dependent. Consequently, the field-dependent AMS enables the magnetic fabric of the latter group of minerals to be separated from the whole-rock AMS. The methods for the determination of the field-dependent AMS consist of separate measurements of each specimen in several fields within the Rayleigh Law range and subsequent processing in which the field-independent and field-dependent AMS components are calculated. The disadvantage of this technique is that each specimen must be measured several times, which is relatively laborious and time consuming. Recently, a new 3D rotator was developed for the MFK1-FA Kappabridge, which rotates the specimen simultaneously about two axes with different velocities. The measurement is fully automated in such a way that, once the specimen is inserted into the rotator, it requires no additional manipulation to measure the full AMS tensor. Consequently, the 3D rotator enables to measure the AMS tensors in the pre-set field intensities without any operator interference. Whole procedure is controlled by newly developed Safyr5 software; once the measurements are finished, the acquired data are immediately processed and can be visualized in a standard way.

  20. Scaled Composites' Proteus aircraft and an F/A-18 Hornet from NASA's Dryden Flight Research Center d

    NASA Technical Reports Server (NTRS)

    2002-01-01

    Scaled Composites' Proteus aircraft and an F/A-18 Hornet from NASA's Dryden Flight Research Center during a low-level flyby at Las Cruces Airport in New Mexico. The unique Proteus aircraft served as a test bed for NASA-sponsored flight tests designed to validate collision-avoidance technologies proposed for uninhabited aircraft. The tests, flown over southern New Mexico in March, 2002, used the Proteus as a surrogate uninhabited aerial vehicle (UAV) while three other aircraft flew toward the Proteus from various angles on simulated collision courses. Radio-based 'detect, see and avoid' equipment on the Proteus successfully detected the other aircraft and relayed that information to a remote pilot on the ground at Las Cruces Airport. The pilot then transmitted commands to the Proteus to maneuver it away from the potential collisions. The flight demonstration, sponsored by NASA Dryden Flight Research Center, New Mexico State University, Scaled Composites, the U.S. Navy and Modern Technology Solutions, Inc., were intended to demonstrate that UAVs can be flown safely and compatibly in the same skies as piloted aircraft.

  1. Determination of the stability and control derivatives of the NASA F/A-18 HARV using flight data

    NASA Technical Reports Server (NTRS)

    Napolitano, Marcello R.; Spagnuolo, Joelle M.

    1993-01-01

    This report documents the research conducted for the NASA-Ames Cooperative Agreement No. NCC 2-759 with West Virginia University. A complete set of the stability and control derivatives for varying angles of attack from 10 deg to 60 deg were estimated from flight data of the NASA F/A-18 HARV. The data were analyzed with the use of the pEst software which implements the output-error method of parameter estimation. Discussions of the aircraft equations of motion, parameter estimation process, design of flight test maneuvers, and formulation of the mathematical model are presented. The added effects of the thrust vectoring and single surface excitation systems are also addressed. The results of the longitudinal and lateral directional derivative estimates at varying angles of attack are presented and compared to results from previous analyses. The results indicate a significant improvement due to the independent control surface deflections induced by the single surface excitation system, and at the same time, a need for additional flight data especially at higher angles of attack.

  2. Liquid filled prismatic louver façade for enhanced daylighting in high-rise commercial buildings.

    PubMed

    Vlachokostas, A; Madamopoulos, N

    2015-07-27

    A liquid filled prismatic louver (LFPL) façade that can perform daylight and thermal energy harvesting with the potential to offer enhanced natural illumination levels to office spaces and thermally assist secondary thermal driven applications is proposed and analyzed. We focus the present simulation study on the evaluation of daylight enhancement in indoor space by redirecting light from a window opening to the ceiling of the room, and then-after a diffusive reflection from the ceiling-onward to the work plane of the room. Illumination simulations using LightTools, a forward ray tracing illumination simulation software, are performed for an office building space located in New York City. We show that the LFPL system achieves deeper natural light penetration, better uniformity and higher illuminance levels compared to an office space without the LFPL system. We further extend our study to a number of other representative cities in the continental US, covering different climatic zones. The LFPL system achieves good daylight harvesting performance. Finally, we discuss the potential of the LFPL system to capture solar infrared radiation heat within the liquid (e.g., water) volume and use it to assist in secondary thermal energy applications. PMID:26367682

  3. Correlates of avian building strikes at a glass façade museum surrounded by avian habitat

    NASA Astrophysics Data System (ADS)

    Kahle, L.; Flannery, M.; Dumbacher, J. P.

    2013-12-01

    Bird window collisions are the second largest anthropogenic cause of bird deaths in the world. Effective mitigation requires an understanding of which birds are most likely to strike, when, and why. Here, we examine five years of avian window strike data from the California Academy of Sciences - a relatively new museum with significant glass façade situated in Golden Gate Park, San Francisco. We examine correlates of window-killed birds, including age, sex, season, and migratory or sedentary tendencies of the birds. We also examine correlates of window kills such as presence of habitat surrounding the building and overall window area. We found that males are almost three times more likely than females to mortally strike windows, and immature birds are three times more abundant than adults in our window kill dataset. Among seasons, strikes were not notably different in spring, summer, and fall; however they were notably reduced in winter. There was no statistical effect of building orientation (north, south, east, or west), and the presence of avian habitat directly adjacent to windows had a minor effect. We also report ongoing studies examining various efforts to reduce window kill (primarily external decals and large electronic window blinds.) We hope that improving our understanding of the causes of the window strikes will help us strategically reduce window strikes.

  4. [Cluster headache differential diagnosis].

    PubMed

    Guégan-Massardier, Evelyne; Laubier, Cécile

    2015-11-01

    Cluster headache is characterized by disabling stereotyped headache. Early diagnosis allows appropriate treatment, unfortunately diagnostic errors are frequent. The main differential diagnoses are other primary or essential headaches. Migraine, more frequent and whose diagnosis is carried by excess, trigeminal neuralgia or other trigemino-autonomic cephalgia. Vascular or tumoral underlying condition can mimic cluster headache, neck and brain imaging is recommended, ideally MRI. PMID:26549687

  5. Wild Duck Cluster

    NASA Technical Reports Server (NTRS)

    2005-01-01

    On April 7, 2005, the Deep Impact spacecraft's Impactor Target Sensor camera recorded this image of M11, the Wild Duck cluster, a galactic open cluster located 6 thousand light years away. The camera is located on the impactor spacecraft, which will image comet Tempel 1 beginning 22 hours before impact until about 2 seconds before impact. Impact with comet Tempel 1 is planned for July 4, 2005.

  6. Synthesis of 3-chloro-6-((4-(di-tert-butyl[(18)F]fluorosilyl)-benzyl)oxy)-1,2,4,5-tetrazine ([(18)F]SiFA-OTz) for rapid tetrazine-based (18)F-radiolabeling.

    PubMed

    Zhu, Jun; Li, Stephen; Wängler, Carmen; Wängler, Björn; Lennox, R Bruce; Schirrmacher, Ralf

    2015-08-11

    An efficient method to prepare the (18)F-labeled tetrazine-derivative [(18)F]-SiFA-OTz for bioorthogonal radiochemistry was developed. [(18)F]-SiFA-OTz can be synthesized with a radiochemical yield of 78 ± 5% within 25 min and can quantitatively react with a model strained dienophile, trans-cyclooctenol. PMID:26145162

  7. Cluster functional renormalization group

    NASA Astrophysics Data System (ADS)

    Reuther, Johannes; Thomale, Ronny

    2014-01-01

    Functional renormalization group (FRG) has become a diverse and powerful tool to derive effective low-energy scattering vertices of interacting many-body systems. Starting from a free expansion point of the action, the flow of the RG parameter Λ allows us to trace the evolution of the effective one- and two-particle vertices towards low energies by taking into account the vertex corrections between all parquet channels in an unbiased fashion. In this work, we generalize the expansion point at which the diagrammatic resummation procedure is initiated from a free UV limit to a cluster product state. We formulate a cluster FRG scheme where the noninteracting building blocks (i.e., decoupled spin clusters) are treated exactly, and the intercluster couplings are addressed via RG. As a benchmark study, we apply our cluster FRG scheme to the spin-1/2 bilayer Heisenberg model (BHM) on a square lattice where the neighboring sites in the two layers form the individual two-site clusters. Comparing with existing numerical evidence for the BHM, we obtain reasonable findings for the spin susceptibility, the spin-triplet excitation energy, and quasiparticle weight even in coupling regimes close to antiferromagnetic order. The concept of cluster FRG promises applications to a large class of interacting electron systems.

  8. Communication: The cluster vapor to cluster solid transition

    NASA Astrophysics Data System (ADS)

    Sweatman, Martin B.; Lue, Leo

    2016-05-01

    Until now, depletion induced transitions have been the hallmark of multicomponent systems only. Monte Carlo simulations reveal a depletion-induced phase transition from cluster vapor to cluster solid in a one-component fluid with competing short range and long range interactions. This confirms a prediction made by earlier theoretical work. Analysis of renormalized cluster-cluster and cluster-vapor interactions suggests that a cluster liquid is also expected within a very narrow range of model parameters. These insights could help identify the mechanisms of clustering in experiments and assist the design of colloidal structures through engineered self-assembly.

  9. The Clustering of Young Stellar Cluster Populations in Nearby Galaxies

    NASA Astrophysics Data System (ADS)

    Grasha, Kathryn; Calzetti, Daniela

    2016-01-01

    We present measurements of clustering among star clusters for several galaxies drawn from the Legacy ExtraGalactic UV Survey (LEGUS), in order to establish whether the clustering strength depends on properties of the cluster population. We use the two point autocorrelation function to study clustering as a function of spatial scale, age, concentration index (CI), and mass. We separate the clusters into different classes, defined by their (a)symmetry and number of peaks, comparing the trends of the autocorrelation functions between all the cluster classes. For one galaxy, NGC 628, we find that younger star clusters are more strongly clustered over small spatial scales and that the clustering disappears rapidly for ages as young as 40 Myr. We present here a similar analysis for the other galaxies. We also measure the power-law slope and amplitude of the autocorrelation functions and discuss the results.

  10. Relation chain based clustering analysis

    NASA Astrophysics Data System (ADS)

    Zhang, Cheng-ning; Zhao, Ming-yang; Luo, Hai-bo

    2011-08-01

    Clustering analysis is currently one of well-developed branches in data mining technology which is supposed to find the hidden structures in the multidimensional space called feature or pattern space. A datum in the space usually possesses a vector form and the elements in the vector represent several specifically selected features. These features are often of efficiency to the problem oriented. Generally, clustering analysis goes into two divisions: one is based on the agglomerative clustering method, and the other one is based on divisive clustering method. The former refers to a bottom-up process which regards each datum as a singleton cluster while the latter refers to a top-down process which regards entire data as a cluster. As the collected literatures, it is noted that the divisive clustering is currently overwhelming both in application and research. Although some famous divisive clustering methods are designed and well developed, clustering problems are still far from being solved. The k - means algorithm is the original divisive clustering method which initially assigns some important index values, such as the clustering number and the initial clustering prototype positions, and that could not be reasonable in some certain occasions. More than the initial problem, the k - means algorithm may also falls into local optimum, clusters in a rigid way and is not available for non-Gaussian distribution. One can see that seeking for a good or natural clustering result, in fact, originates from the one's understanding of the concept of clustering. Thus, the confusion or misunderstanding of the definition of clustering always derives some unsatisfied clustering results. One should consider the definition deeply and seriously. This paper demonstrates the nature of clustering, gives the way of understanding clustering, discusses the methodology of designing a clustering algorithm, and proposes a new clustering method based on relation chains among 2D patterns. In

  11. One-step ¹⁸F-labeling of carbohydrate-conjugated octreotate-derivatives containing a silicon-fluoride-acceptor (SiFA): in vitro and in vivo evaluation as tumor imaging agents for positron emission tomography (PET).

    PubMed

    Wängler, Carmen; Waser, Beatrice; Alke, Andrea; Iovkova, Ljuba; Buchholz, Hans-Georg; Niedermoser, Sabrina; Jurkschat, Klaus; Fottner, Christian; Bartenstein, Peter; Schirrmacher, Ralf; Reubi, Jean-Claude; Wester, Hans-Jürgen; Wängler, Björn

    2010-12-15

    The synthesis, radiolabeling, and initial evaluation of new silicon-fluoride acceptor (SiFA) derivatized octreotate derivatives is reported. So far, the main drawback of the SiFA technology for the synthesis of PET-radiotracers is the high lipophilicity of the resulting radiopharmaceutical. Consequently, we synthesized new SiFA-octreotate analogues derivatized with Fmoc-NH-PEG-COOH, Fmoc-Asn(Ac₃AcNH-β-Glc)-OH, and SiFA-aldehyde (SIFA-A). The substances could be labeled in high yields (38 ± 4%) and specific activities between 29 and 56 GBq/μmol in short synthesis times of less than 30 min (e.o.b.). The in vitro evaluation of the synthesized conjugates displayed a sst2 receptor affinity (IC₅₀ = 3.3 ± 0.3 nM) comparable to that of somatostatin-28. As a measure of lipophilicity of the conjugates, the log P(ow) was determined and found to be 0.96 for SiFA-Asn(AcNH-β-Glc)-PEG-Tyr³-octreotate and 1.23 for SiFA-Asn(AcNH-β-Glc)-Tyr³-octreotate, which is considerably lower than for SiFA-Tyr³-octreotate (log P(ow) = 1.59). The initial in vivo evaluation of [¹⁸F]SiFA-Asn(AcNH-β-Glc)-PEG-Tyr³-octreotate revealed a significant uptake of radiotracer in the tumor tissue of AR42J tumor-bearing nude mice of 7.7% ID/g tissue weight. These results show that the high lipophilicity of the SiFA moiety can be compensated by applying hydrophilic moieties. Using this approach, a tumor-affine SiFA-containing peptide could successfully be used for receptor imaging for the first time in this proof of concept study. PMID:21082773

  12. Friedreich's Ataxia (FA)

    MedlinePlus

    ... About Friedreich’s Ataxia Updated December 2009 Michelle Moffitt Smith Michelle and James Smith at their wedding Dear Friends: W hen I ... for a doctorate. I met my husband, James Smith, through MDA’s magazine, Quest. He also has a ...

  13. Installed F/A-18 inlet flow calculations at 30 degrees angle-of-attack: A comparative study

    NASA Technical Reports Server (NTRS)

    Smith, C. Frederic; Podleski, Steve D.

    1994-01-01

    NASA Lewis is currently engaged in a research effort as a team member of the High Alpha Technology Program (HATP) within NASA. This program utilizes a specially equipped F/A-18, the High Alpha Research Vehicle (HARV), in an ambitious effort to improve the maneuverability of high-performance military aircraft at low subsonic speed, high angle of attack conditions. The overall objective of the Lewis effort is to develop inlet technology that will ensure efficient airflow delivery to the engine during these maneuvers. One part of the Lewis approach utilizes computational fluid dynamics codes to predict the installed performance of inlets for these highly maneuverable aircraft. Full Navier-Stokes (FNS) calculations on the installed F/A-18 inlet at 30 degrees angle of attack, 0 degrees yaw, and a freestream Mach number of 0.2 have been obtained in this study using an algebraic turbulence model with two grids (original and revised). Results obtained with the original grid were used to determine where further grid refinements and additional geometry were needed. In order to account properly for the external effects, the forebody, leading edge extension (LEX), ramp, and wing were included with inlet geometry. In the original grid, the diverter, LEX slot, and leading edge flap were not included due to insufficient geometry definition, but were included in a revised grid. In addition, a thin-layer Navier-Stokes (TLNS) code is used with the revised grid and the numerical results are compared to those obtained with the FNS code. The TLNS code was used to evaluate the effects on the solution using a code with more recent CFD developments such as upwinding with TVD schemes versus central differencing with artificial dissipation. The calculations are compared to a limited amount of available experimental data. The predicted forebody/fuselage surface static pressures compared well with data of all solutions. The predicted trajectory of the vortex generated under the LEX was

  14. Clustering in bubbly liquids

    NASA Astrophysics Data System (ADS)

    Figueroa, Bernardo; Zenit, Roberto

    2004-11-01

    We are conducting experiments to determine the amount of clustering that occurs when small gas bubbles ascend in clean water. In particular, we are interested in flows for which the liquid motion around the bubbles can be described, with a certain degree of accuracy, using potential flow theory. This model is applicable for the case of bubbly liquids in which the Reynolds number is large and the Weber number is small. To clearly observe the formation of bubble clusters we propose the use of a Hele-Shaw-type channel. In this thin channel the bubbles cannot overlap in the depth direction, therefore the identification of bubble clusters cannot be misinterpreted. Direct video image analysis is performed to calculate the velocity and size of the bubbles, as well as the formation of clusters. Although the walls do affect the motion of the bubbles, the clustering phenomena does occur and has the same qualitative behavior as in fully three-dimensional flows. A series of preliminary measurements are presented. A brief discussion of our plans to perform PIV measurements to obtain the liquid velocity fields is also presented.

  15. HST Cluster Supernova Survey

    NASA Astrophysics Data System (ADS)

    Suzuki, Nao; Aldering, G.; Amanullah, R.; Barbary, K.; Barrientos, L.; Brodwin, M.; Connolly, N.; Dawson, K.; de Jong, R.; Dey, A.; Doi, M.; Donahue, M.; Eisenhardt, P.; Ellingson, E.; Faccioli, L.; Fadeyev, V.; Fakhouri, H.; Fruchter, A.; Gilbank, D.; Gladders, M.; Goldhaber, G.; Gonzalez, A.; Goobar, A.; Gude, A.; Hennawi, J.; Hoekstra, H.; Hsiao, E.; Huang, X.; Ihara, Y.; Jannuzi, B.; Jee, M. J.; Koester, B.; Kowalski, M.; Lidman, C.; Linder, E.; Lubin, L.; Morokuma, T.; Perlmutter, S.; Postman, M.; Rhodes, J.; Rosati, P.; Ripoche, P.; Rubin, D.; Schlegel, D.; Spadafora, A.; Stanford, A.; Stern, D.; Yasuda, N.; Yee, H.; Cosmology Project, Supernova

    2010-01-01

    We report results from the Hubble Space Telescope (HST) Cluster Supernova Survey with the Advanced Camera for Surveys (ACS) (PI: Perlmutter; see Dawson et al. AJ, 2009). We have introduced a novel approach to discover and follow Type Ia supernovae (SNeIa). With HST, we monitored 25 massive clusters (0.9 < z < 1.4) found by the RCS, XMM, IRAC, and RDCS surveys and conducted spectroscopic observations with the Keck, Subaru, and VLT telescopes. Sixteen SNe were discovered at 0.95 < z < 1.41, nine of which were in galaxy clusters (for a discussion of the rates, see K. Barbary, oral presentation at this meeting). The SNe in galaxy clusters are found primarily in early type galaxies in the cluster red-sequence that have been shown to be nearly dust-free and uniform populations (see poster presentation by Meyers et al.). The reduction and control of systematic error is an urgent task for the study of dark energy today (see Rubin et al. poster presentation), and we discuss how this unique SNe Ia data set reduces both statistical and more importantly systematic uncertainty at the highest redshifts. This work has been supported by the Office of Science, U.S. Department of Energy, through contract DE-AC02-05CH11231 and in part by NASA through grants associated with HST-GO-10496.

  16. ROTATING GLOBULAR CLUSTERS

    SciTech Connect

    Bianchini, P.; Varri, A. L.; Bertin, G.; Zocchi, A.

    2013-07-20

    Internal rotation is thought to play a major role in the dynamics of some globular clusters. However, in only a few cases has internal rotation been studied by the quantitative application of realistic and physically justified global models. Here, we present a dynamical analysis of the photometry and three-dimensional kinematics of {omega} Cen, 47 Tuc, and M15, by means of a recently introduced family of self-consistent axisymmetric rotating models. The three clusters, characterized by different relaxation conditions, show evidence of differential rotation and deviations from sphericity. The combination of line-of-sight velocities and proper motions allows us to determine their internal dynamics, predict their morphology, and estimate their dynamical distance. The well-relaxed cluster 47 Tuc is interpreted very well by our model; internal rotation is found to explain the observed morphology. For M15, we provide a global model in good agreement with the data, including the central behavior of the rotation profile and the shape of the ellipticity profile. For the partially relaxed cluster {omega} Cen, the selected model reproduces the complex three-dimensional kinematics; in particular, the observed anisotropy profile, characterized by a transition from isotropy to weakly radial anisotropy and then to tangential anisotropy in the outer parts. The discrepancy found for the steep central gradient in the observed line-of-sight velocity dispersion profile and for the ellipticity profile is ascribed to the condition of only partial relaxation of this cluster and the interplay between rotation and radial anisotropy.

  17. Wind tunnel investigations of forebody strakes for yaw control on F/A-18 model at subsonic and transonic speeds

    NASA Technical Reports Server (NTRS)

    Erickson, Gary E.; Murri, Daniel G.

    1993-01-01

    Wind tunnel investigations have been conducted of forebody strakes for yaw control on 0.06-scale models of the F/A-18 aircraft at free-stream Mach numbers of 0.20 to 0.90. The testing was conducted in the 7- by 10-Foot Transonic Tunnel at the David Taylor Research Center and the Langley 7- by 10-Foot High-Speed Tunnel. The principal objectives of the testing were to determine the effects of the Mach number and the strake plan form on the strake yaw control effectiveness and the corresponding strake vortex induced flow field. The wind tunnel model configurations simulated an actuated conformal strake deployed for maximum yaw control at high angles of attack. The test data included six-component forces and moments on the complete model, surface static pressure distributions on the forebody and wing leading-edge extensions, and on-surface and off-surface flow visualizations. The results from these studies show that the strake produces large yaw control increments at high angles of attack that exceed the effect of conventional rudders at low angles of attack. The strake yaw control increments diminish with increasing Mach number but continue to exceed the effect of rudder deflection at angles of attack greater than 30 degrees. The character of the strake vortex induced flow field is similar at subsonic and transonic speeds. Cropping the strake planform to account for geometric and structural constraints on the F-18 aircraft has a small effect on the yaw control increments at subsonic speeds and no effect at transonic speeds.

  18. Yamadazyma ubonensis f.a., sp. nov., a novel xylitol-producing yeast species isolated in Thailand.

    PubMed

    Junyapate, Keeratichaiyanan; Jindamorakot, Sasitorn; Limtong, Savitree

    2014-03-01

    Three hundred and thirty-seven xylose-utilizing yeast strains were isolated from various natural samples. Among these, 68 strains produced xylitol in the range of 0.1-0.69 g xylitol/g xylose. Thirty-nine xylitol-producing strains were identified to be Candida tropicalis. Ten strains were found belonging to 14 known species in the genus Candida, Cyberlindnera, Meyerozyma, Pichia, Wickerhamomyces, Yamadazyma and Cryptococcus. Two strains were identified to be two Candida species and two strains (DMKU-XE142(T) and DMKU-XE332) were found to be a novel species. Strain DMKU-XE142(T) was isolated from tree bark and DMKU-XE332 was obtained from decaying plant leaf collected in Thailand. On the basis of morphological, biochemical, physiological and chemotaxonomic characteristics and sequence analysis of the D1/D2 region of the large subunit rRNA gene (LSU) and the internal transcribed spacer (ITS) region, the two strains were determined to represent a novel Yamadazyma species although formation of ascospores was not observed. The sequences of the D1/D2 region of the LSU rRNA gene and the ITS region of the two strains were identical but differed from Yamadazyma phyllophila, the closest species in terms of pairwise sequence similarity of the D1/D2 region, by 1.7 % nucleotide substitutions and 3.5 % nucleotide substitutions in the ITS region. The name Yamadazyma ubonensis f.a., sp. nov. is proposed (type strain is DMKU-XE142(T) = BCC 61020(T) = CBS 12859(T)). PMID:24346846

  19. Neutralization of Antimicrobial Substances in New BacT/Alert FA and FN Plus Blood Culture Bottles

    PubMed Central

    Barousch, Wolfgang; Nehr, Marion; Kundi, Michael; Zeitlinger, Markus; Makristathis, Athanasios; Hirschl, Alexander M.

    2013-01-01

    Time to detection (TTD) in automated blood culture systems is delayed for sensitive microorganisms in the presence of antimicrobial substances and has been associated with worse outcomes for sepsis patients on inadequate empirical therapy. While resin addition removes antimicrobial substances to various degrees from blood culture media, media formulations and the blend of resins may influence performance. The BacT/Alert 3D system (bioMérieux) was investigated using the new resin-containing medium types FA Plus (aerobic) and FN Plus (anaerobic). TTD was compared between control and test bottles containing relevant bacteria or Candida albicans, with and without defined concentrations of antimicrobials. Failure of neutralization was defined as a negative blood culture on day 3. In general, growth delay was nonlinear, concentration dependent, bottle type specific, and reciprocally associated with MICs. Substance-specific serum drug concentrations corresponding to a predefined, clinically relevant 3-h delay of TTD were calculated. Where appropriate, a time interval allowing for drug elimination below this critical level was obtained by pharmacokinetic modeling. Clarithromycin, clindamycin, gentamicin, linezolid, tigecycline, vancomycin, and fluconazole were neutralized. For ciprofloxacin and piperacillin-tazobactam, which were only incompletely neutralized in combination with the most sensitive test strains, a maximum waiting time for blood draw of 1 h was determined based on pharmacokinetics. One or more test strains did not grow in bottles containing either amoxicillin-clavulanate, cefepime, cefotaxime, meropenem, or metronidazole, and we thus recommend particular caution in timing of blood draws if patients have been pretreated with these agents. PMID:23486710

  20. Functional characterization of a PEI-CyD-FA-coated adenovirus as delivery vector for gene therapy.

    PubMed

    Yao, Hong; Chen, Shih-Chi; Shen, Zan; Huang, Yun-Chao; Zhu, Xiao; Wang, Xiao-mei; Jiang, Wenqi; Wang, Zi-Feng; Bian, Xiu-Wu; Ling, Eng-Ang; Kung, Hsiang-fu; Lin, Marie C

    2013-01-01

    The recombinant adenovirus is evolving as a promising gene delivery vector for gene therapy due to its efficiency in transducing different genes into most types of cells. However, the host-immune response elicited by primary inoculation of an adenovirus can cause rapid clearance of the vector, impairing the efficacy of the adenovirus and hence obstructing its clinical application. We have previously synthesized a biodegradable co-polymer consisting of a low molecular weight PEI (MW 600 Da), cross-linked with β-cyclodextrin, and conjugated with folic acid (PEI-CyD-FA, named H1). Here we report that coating the adenovirus vector (Adv) with H1 (H1/rAdv) could significantly improve both the efficacy and biosafety of Adv. Enhanced transfection efficiency as well as prolonged duration of gene expression were clearly demonstrated either by intratumoral or systemic injection of a single dose of H1/rAdv in immunocompetent mice. Importantly, repeated injections of H1/rAdv did not reduce the transfection efficiency in immunocompetent mice. Furthermore, H1 transformed the surface charge of the adenovirus capsomers from negative to positive in physiological solution, suggesting that H1 coated the capsid protein of the adenovirus. This could shelter the epitopes of capsid proteins of the adenovirus, resulting in a reduced host-immune response and enhanced transfection efficiency. Taken together, these findings suggest that H1/rAdv is an effective gene delivery system superior to the adenovirus alone and that it could be considered as a preferred vehicle for gene therapy. PMID:23531212