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Sample records for abasic site ap

  1. Structure and function of the abasic site specificity pocket of an AP endonuclease from Archaeoglobus fulgidus.

    PubMed

    Schmiedel, Ramona; Kuettner, E Bartholomeus; Keim, Antje; Sträter, Norbert; Greiner-Stöffele, Thomas

    2009-02-01

    The major AP endonuclease in Escherichia coli Exonuclease III (ExoIII) is frequently used in gene technology due to its strong exonucleolytic activity. A thermostabilized variant of ExoIII or a homologous enzyme from thermophilic organisms could be most useful for further applications. For this purpose we characterized a nuclease from the hyperthermophilic archaeon Archaeoglobus fulgidus (Af_Exo), which shares 33% overall sequence identity and 55% similarity to ExoIII. The gene coding for this thermostable enzyme was cloned and expressed in E. coli. The purified protein shows a strong Mg(2+)-dependent nicking activity at AP-sites, nicking of undamaged double-stranded (ds) DNA and a weak exonucleolytic activity. A V217G variant of the enzyme was crystallized with decamer ds-DNA molecule, and the three-dimensional structure was determined to 1.7A resolution. Besides our goal to find or produce a thermostable exonuclease, the structural and catalytic data of Af_Exo and a series of mutant proteins, based on the crystal structure, provide new insight into the mechanism of abasic site recognition and repair. Each of the hydrophobic residues Phe 200, Trp 215 and Val 217, forming a binding pocket for the abasic deoxyribose in Af_Exo, were mutated to glycine or serine. By expanding the size of the binding pocket the unspecific endonucleolytic activity is increased. Thus, size and flexibility of the mostly hydrophobic binding pocket have a significant influence on AP-site specificity. We suggest that its tight fitting to the flipped-out deoxyribose allows for a preferred competent binding of abasic sites. In a larger or more flexible pocket however, intact nucleotides more easily bind in a catalytically competent conformation, resulting in loss of specificity. Moreover, with mutations of Phe 200 and Trp 215 we induced a strong exonucleolytic activity on undamaged DNA. PMID:19015049

  2. Shape-selective recognition of DNA abasic sites by metallohelices: inhibition of human AP endonuclease 1

    PubMed Central

    Malina, Jaroslav; Scott, Peter; Brabec, Viktor

    2015-01-01

    Loss of a base in DNA leading to creation of an abasic (AP) site leaving a deoxyribose residue in the strand, is a frequent lesion that may occur spontaneously or under the action of various physical and chemical agents. Progress in the understanding of the chemistry and enzymology of abasic DNA largely relies upon the study of AP sites in synthetic duplexes. We report here on interactions of diastereomerically pure metallo–helical ‘flexicate’ complexes, bimetallic triple-stranded ferro-helicates [Fe2(NN-NN)3]4+ incorporating the common NN–NN bis(bidentate) helicand, with short DNA duplexes containing AP sites in different sequence contexts. The results show that the flexicates bind to AP sites in DNA duplexes in a shape-selective manner. They preferentially bind to AP sites flanked by purines on both sides and their binding is enhanced when a pyrimidine is placed in opposite orientation to the lesion. Notably, the Λ-enantiomer binds to all tested AP sites with higher affinity than the Δ-enantiomer. In addition, the binding of the flexicates to AP sites inhibits the activity of human AP endonuclease 1, which is as a valid anticancer drug target. Hence, this finding indicates the potential of utilizing well-defined metallo–helical complexes for cancer chemotherapy. PMID:25940617

  3. Abasic and oxidized abasic site reactivity in DNA: enzyme inhibition, cross-linking, and nucleosome catalyzed reactions.

    PubMed

    Greenberg, Marc M

    2014-02-18

    Abasic lesions are a family of DNA modifications that lack Watson-Crick bases. The parent member of this family, the apurinic/apyrimidinic lesion (AP), occurs as an intermediate during DNA repair, following nucleobase alkylation, and from random hydrolysis of native nucleotides. In a given day, each cell produces between 10000 and 50000 AP lesions. A variety of oxidants including γ-radiolysis produce oxidized abasic sites, such as C4-AP, from the deoxyribose backbone. A number of potent, cytotoxic antitumor agents, such as bleomycin and the enediynes (e.g., calicheamicin, esperamicin, and neocarzinostatin) also lead to oxidized abasic sites in DNA. The absence of Watson-Crick bases prevents DNA polymerases from properly determining which nucleotide to incorporate opposite abasic lesions. Consequently, several studies have revealed that (oxidized) abasic sites are highly mutagenic. Abasic lesions are also chemically unstable, are prone to strand scission, and possess electrophilic carbonyl groups. However, researchers have only uncovered the consequences of the inherent reactivity of these electrophiles within the past decade. The development of solid phase synthesis methods for oligonucleotides that both place abasic sites in defined positions and circumvent their inherent alkaline lability has facilitated this research. Chemically synthesized oligonucleotides containing abasic lesions provide substrates that have allowed researchers to discover a range of interesting chemical properties of potential biological importance. For instance, abasic lesions form DNA-DNA interstrand cross-links, a particularly important family of DNA damage because they block replication and transcription absolutely. In addition, bacterial repair enzymes can convert an interstrand cross-link derived from C4-AP into a double-strand break, the most deleterious form of DNA damage. Oxidized abasic lesions can also inhibit DNA repair enzymes that remove damaged nucleotides. DNA polymerase

  4. DNA Polymerase λ Inactivation by Oxidized Abasic Sites&

    PubMed Central

    Stevens, Adam J.; Guan, Lirui; Bebenek, Katarzyna; Kunkel, Thomas A.; Greenberg, Marc M.

    2013-01-01

    Base excision repair plays a vital role in maintaining genomic integrity in mammalian cells. DNA polymerase λ is believed to play a backup role to DNA polymerase β in base excision repair. Two oxidized abasic lesions that are produced by a variety of DNA damaging agents, including several antitumor antibiotics, the C4′-oxidized abasic site following Ape1 incision (pC4-AP) and 5′-(2-phosphoryl-1,4-dioxobutane) (DOB), irreversibly inactivate Pol β and Pol λ. The interactions of DOB and pC4-AP with Pol λ are examined in detail using DNA substrates containing these lesions at defined sites. Single turnover kinetic experiments show that Pol λ excises DOB almost 13-times more slowly than a 5′-phosphorylated 2-deoxyribose (dRP). pC4-AP is excised approximately twice as fast as DOB. The absolute rate constants are considerably slower than those reported for Pol β at the respective reactions, suggesting that Pol λ may be an inefficient backup in BER. DOB inactivates Pol λ approximately 3-fold less efficiently than it does Pol β and the difference is attributable to a higher KI (33 ± 7 nM). Inactivation of Pol λ’s lyase activity by DOB also prevents the enzyme from carrying out polymerization following preincubation of the protein and DNA. Mass spectral analysis of GluC digested Pol λ inactivated by DOB shows that Lys324 is modified. There is inferential support that Lys312 may also be modified. Both residues are within the Pol λ lyase active site. Protein modification involves reaction with released but-2-ene-1,4-dial. When acting on pC4-AP, Pol λ achieves approximately 4 turnovers on average before being inactivated. Lyase inactivation by pC4-AP is also accompanied by loss of polymerase activity and mass spectrometry indicates that Lys312 and Lys324 are modified by the lesion. The ability of DOB and pC4-AP to inactivate Pol λ provides additional evidence that these lesions are significant sources of the cytotoxicity of DNA damaging agents that

  5. The Electronic Influence of Abasic Sites in DNA.

    PubMed

    McWilliams, Marc A; Bhui, Rita; Taylor, David W; Slinker, Jason D

    2015-09-01

    Abasic sites in DNA are prevalent as both naturally forming defects and as synthetic inclusions for biosensing applications. The electronic impact of these defects in DNA sensor and device configurations has yet to be clarified. Here we report the effect of an abasic site on the rate and yield of charge transport through temperature-controlled analysis of DNA duplex monolayers on multiplexed devices. Transport yield through the abasic site monolayer strongly increases with temperature, but the yield relative to an undamaged monolayer decreases with temperature. This is opposite to the increasing relative yield with temperature from a mismatched base pair, and these effects are accounted for by the unique structural impact of each defect. Notably, the effect of the abasic site is nearly doubled when heated from room temperature to 37 °C. The rate of transport is largely unaffected by the abasic site, showing Arrhenius-type behavior with an activation energy of ∼300 meV. Detailed abasic site investigation elucidates the electrical impact of these biologically spontaneous defects and aids development of biological sensors. PMID:26280191

  6. Conformational dynamics of abasic DNA upon interactions with AP endonuclease 1 revealed by stopped-flow fluorescence analysis.

    PubMed

    Kanazhevskaya, Lyubov Yu; Koval, Vladimir V; Vorobjev, Yury N; Fedorova, Olga S

    2012-02-14

    Apurinic/apyrimidinic (AP) sites are abundant DNA lesions arising from exposure to UV light, ionizing radiation, alkylating agents, and oxygen radicals. In human cells, AP endonuclease 1 (APE1) recognizes this mutagenic lesion and initiates its repair via a specific incision of the phosphodiester backbone 5' to the AP site. We have investigated a detailed mechanism of APE1 functioning using fluorescently labeled DNA substrates. A fluorescent adenine analogue, 2-aminopurine, was introduced into DNA substrates adjacent to the abasic site to serve as an on-site reporter of conformational transitions in DNA during the catalytic cycle. Application of a pre-steady-state stopped-flow technique allows us to observe changes in the fluorescence intensity corresponding to different stages of the process in real time. We also detected an intrinsic Trp fluorescence of the enzyme during interactions with 2-aPu-containing substrates. Our data have revealed a conformational flexibility of the abasic DNA being processed by APE1. Quantitative analysis of fluorescent traces has yielded a minimal kinetic scheme and appropriate rate constants consisting of four steps. The results obtained from stopped-flow data have shown a substantial influence of the 2-aPu base location on completion of certain reaction steps. Using detailed molecular dynamics simulations of the DNA substrates, we have attributed structural distortions of AP-DNA to realization of specific binding, effective locking, and incision of the damaged DNA. The findings allowed us to accurately discern the step that corresponds to insertion of specific APE1 amino acid residues into the abasic DNA void in the course of stabilization of the precatalytic complex. PMID:22243137

  7. Synthesis of Cross-Linked DNA Containing Oxidized Abasic Site Analogues

    PubMed Central

    2015-01-01

    DNA interstrand cross-links are an important family of DNA damage that block replication and transcription. Recently, it was discovered that oxidized abasic sites react with the opposing strand of DNA to produce interstrand cross-links. Some of the cross-links between 2′-deoxyadenosine and the oxidized abasic sites, 5′-(2-phosphoryl-1,4-dioxobutane) (DOB) and the C4-hydroxylated abasic site (C4-AP), are formed reversibly. Chemical instability hinders biochemical, structural, and physicochemical characterization of these cross-linked duplexes. To overcome these limitations, we developed methods for preparing stabilized analogues of DOB and C4-AP cross-links via solid-phase oligonucleotide synthesis. Oligonucleotides of any sequence are attainable by synthesizing phosphoramidites in which the hydroxyl groups of the cross-linked product were orthogonally protected using photochemically labile and hydrazine labile groups. Selective unmasking of a single hydroxyl group precedes solid-phase synthesis of one arm of the cross-linked DNA. The method is compatible with commercially available phosphoramidites and other oligonucleotide synthesis reagents. Cross-linked duplexes containing as many as 54 nt were synthesized on solid-phase supports. Subsequent enzyme ligation of one cross-link product provided a 60 bp duplex, which is suitable for nucleotide excision repair studies. PMID:24949656

  8. Recognition of DNA abasic site nanocavity by fluorophore-switched probe: Suitable for all sequence environments

    NASA Astrophysics Data System (ADS)

    Wang, Ying; Hu, Yuehua; Wu, Tao; Zhang, Lihua; Liu, Hua; Zhou, Xiaoshun; Shao, Yong

    2016-01-01

    Removal of a damaged base in DNA produces an abasic site (AP site) nanocavity. If left un-repaired in vivo by the specific enzyme, this nanocavity will result in nucleotide mutation in the following DNA replication. Therefore, selective recognition of AP site nanocavity by small molecules is important for identification of such DNA damage and development of genetic drugs. In this work, we investigate the fluorescence behavior of isoquinoline alkaloids including palmatine (PAL), berberine (BER), epiberberine (EPI), jatrorrhizine (JAT), coptisine (COP), coralyne (COR), worenine (WOR), berberrubine (BEU), sanguinarine (SAN), chelerythrine (CHE), and nitidine (NIT) upon binding with the AP nanocavity. PAL is screened out as the most efficient fluorophore-switched probe to recognize the AP nanocavity over the fully matched DNA. Its fluorescence enhancement occurs for all of the AP nanocavity sequence environments, which has not been achieved by the previously used probes. The bridged π conjugation effect should partially contribute to the AP nanocavity-specific fluorescence, as opposed to the solvent effect. Due to the strong binding with the AP nanocavity, PAL will find wide applications in the DNA damage recognition and sensor development.

  9. DNA abasic site-directed formation of fluorescent silver nanoclusters for selective nucleobase recognition

    NASA Astrophysics Data System (ADS)

    Ma, Kun; Cui, Qinghua; Liu, Guiying; Wu, Fei; Xu, Shujuan; Shao, Yong

    2011-07-01

    DNA single-nucleotide polymorphism (SNP) detection has attracted much attention due to mutation related diseases. Various methods for SNP detection have been proposed and many are already in use. Here, we find that the abasic site (AP site) in the DNA duplex can be developed as a capping scaffold for the generation of fluorescent silver nanoclusters (Ag NCs). As a proof of concept, the DNA sequences from fragments near codon 177 of cancer supression gene p53 were used as a model for SNP detection by in situ formed Ag NCs. The formation of fluorescent Ag NCs in the AP site-containing DNA duplex is highly selective for cytosine facing the AP site and guanines flanking the site and can be employed in situ as readout for SNP detection. The fluorescent signal-on sensing for SNP based on this inorganic fluorophore is substantially advantageous over the previously reported signal-off responses using low-molecular-weight organic ligands. The strong dependence of fluorescent Ag NC formation on the sequences surrounding the AP site was successfully used to identify mutations in codon 177 of cancer supression gene p53. We anticipate that this approach will be employed to develop a practical SNP detection method by locating an AP site toward the midway cytosine in a target strand containing more than three consecutive cytosines.

  10. Base excision repair enzymes protect abasic sites in duplex DNA from interstrand cross-links.

    PubMed

    Admiraal, Suzanne J; O'Brien, Patrick J

    2015-03-10

    Hydrolysis of the N-glycosyl bond between a nucleobase and deoxyribose leaves an abasic site within duplex DNA. The abasic site can react with exocyclic amines of nucleobases on the complementary strand to form interstrand DNA-DNA cross-links (ICLs). We find that several enzymes from the base excision repair (BER) pathway protect an abasic site on one strand of a DNA duplex from cross-linking with an amine on the opposing strand. Human alkyladenine DNA glycosylase (AAG) and Escherichia coli 3-methyladenine DNA glycosylase II (AlkA) accomplish this by binding tightly to the abasic site and sequestering it. AAG protects an abasic site opposite T, the product of its canonical glycosylase reaction, by a factor of ∼10-fold, as estimated from its inhibition of the reaction of an exogenous amine with the damaged DNA. Human apurinic/apyrimidinic site endonuclease 1 and E. coli endonuclease III both decrease the amount of ICL at equilibrium by generating a single-strand DNA nick at the abasic position as it is liberated from the cross-link. The reversibility of the reaction between amines and abasic sites allows BER enzymes to counter the potentially disruptive effects of this type of cross-link on DNA transactions. PMID:25679877

  11. The determination of the DNA sequence specificity of bleomycin-induced abasic sites.

    PubMed

    Chen, Jon K; Murray, Vincent

    2016-06-01

    The DNA sequence specificity of the cancer chemotherapeutic agent, bleomycin, was determined with high precision in purified plasmid DNA using an improved technique. This improved technique involved the labelling of the 5'- and 3'-ends of DNA with different fluorescent tags, followed by simultaneous cleavage by bleomycin and capillary electrophoresis with laser-induced fluorescence. This permitted the determination of bleomycin cleavage specificity with high accuracy since end-label bias was greatly reduced. Bleomycin produces single- and double-strand breaks, abasic sites and other base damage in DNA. This high-precision method was utilised to elucidate, for the first time, the DNA sequence specificity of bleomycin-induced DNA damage at abasic sites. This was accomplished using endonuclease IV that cleaves DNA at abasic sites after bleomycin damage. It was found that bleomycin-induced abasic sites formed at 5'-GC and 5'-GT sites while bleomycin-induced phosphodiester strand breaks formed mainly at 5'-GT dinucleotides. Since bleomycin-induced abasic sites are produced in the absence of molecular oxygen, this difference in DNA sequence specificity could be important in hypoxic tumour cells. PMID:26940956

  12. ssDNA Pairing Accuracy Increases When Abasic Sites Divide Nucleotides into Small Groups

    PubMed Central

    Peacock-Villada, Alexandra; Coljee, Vincent; Danilowicz, Claudia; Prentiss, Mara

    2015-01-01

    Accurate sequence dependent pairing of single-stranded DNA (ssDNA) molecules plays an important role in gene chips, DNA origami, and polymerase chain reactions. In many assays accurate pairing depends on mismatched sequences melting at lower temperatures than matched sequences; however, for sequences longer than ~10 nucleotides, single mismatches and correct matches have melting temperature differences of less than 3°C. We demonstrate that appropriately grouping of 35 bases in ssDNA using abasic sites increases the difference between the melting temperature of correct bases and the melting temperature of mismatched base pairings. Importantly, in the presence of appropriately spaced abasic sites mismatches near one end of a long dsDNA destabilize the annealing at the other end much more effectively than in systems without the abasic sites, suggesting that the dsDNA melts more uniformly in the presence of appropriately spaced abasic sites. In sum, the presence of appropriately spaced abasic sites allows temperature to more accurately discriminate correct base pairings from incorrect ones. PMID:26115175

  13. Tandem mass spectrometry-based detection of c4'-oxidized abasic sites at specific positions in DNA fragments.

    PubMed

    Chowdhury, Goutam; Guengerich, F Peter

    2009-07-01

    Oxidative damage to DNA has been linked to aging, cancer, and other biological processes. Reactive oxygen species and various antitumor agents including bleomycin and ionizing radiation have been shown to cause oxidative DNA sugar damage. Detection of DNA lesions is important for understanding the toxicological or therapeutic consequences associated with such agents. C4'-oxidized abasic sites (C4-AP) are produced by the antitumor drug bleomycin and ionizing radiation. The currently available methods for the detection of C4-AP cannot provide both structural and sequence information. We have developed an LC-ESI-MS-based approach for specific detection and mapping of C4-AP from a mixture of lesions. We show using Fe-bleomycin-damaged DNA that C4-AP can be detected at cytosine and thymine sites by direct MS analysis. Our results reveal that collision-induced dissociation of C4-AP-containing oligonucleotides results in preferential fragmentation at C4-AP sites with the formation of the unique a* ions (18 amu more than the a-B ions) that allow mapping of the C4-AP sites. Various chemical modification strategies (e.g., reduction with NaBH4 and NaBD4 and derivatization with methoxyamine and hydrazine, followed by LC-MS analysis) were also used for unambiguous detection of C4-AP sites. Finally, we show that the methods described here can detect the presence of C4-AP at specific sites in a complex sample such as hydroxyl radical-damaged DNA. The LC-MS approach was also used for the simultaneous detection of the other C4'-oxidation end product, 3'-phosphoglycolate, at a specific site in hydroxyl radical-damaged DNA. Thus, LC-MS provides a rapid and direct approach for the detection and mapping of oxidative DNA lesions. PMID:19496605

  14. Triplex molecular beacons for sensitive recognition of melamine based on abasic-site-containing DNA and fluorescent silver nanoclusters.

    PubMed

    Wang, Ya; Sun, Qianqian; Zhu, Linling; Zhang, Junying; Wang, Fengyang; Lu, Linlin; Yu, Haijun; Xu, Zhiai; Zhang, Wen

    2015-05-01

    A melamine aptamer derived from an abasic-site-containing triplex molecular beacon (tMB) was designed and developed for sensitive recognition of melamine by integrating tMBs and fluorescent silver nanoclusters (Ag NCs). PMID:25865656

  15. Accumulation of abasic sites induces genomic instability in normal human gastric epithelial cells during Helicobacter pylori infection.

    PubMed

    Kidane, D; Murphy, D L; Sweasy, J B

    2014-01-01

    Helicobacter pylori infection of the human stomach is associated with inflammation that leads to the release of reactive oxygen and nitrogen species (RONs), eliciting DNA damage in host cells. Unrepaired DNA damage leads to genomic instability that is associated with cancer. Base excision repair (BER) is critical to maintain genomic stability during RONs-induced DNA damage, but little is known about its role in processing DNA damage associated with H. pylori infection of normal gastric epithelial cells. Here, we show that upon H. pylori infection, abasic (AP) sites accumulate and lead to increased levels of double-stranded DNA breaks (DSBs). In contrast, downregulation of the OGG1 DNA glycosylase decreases the levels of both AP sites and DSBs during H. pylori infection. Processing of AP sites during different phases of the cell cycle leads to an elevation in the levels of DSBs. Therefore, the induction of oxidative DNA damage by H. pylori and subsequent processing by BER in normal gastric epithelial cells has the potential to lead to genomic instability that may have a role in the development of gastric cancer. Our results are consistent with the interpretation that precise coordination of BER processing of DNA damage is critical for the maintenance of genomic stability. PMID:25417725

  16. Chemical and structural characterization of interstrand cross-links formed between abasic sites and adenine residues in duplex DNA

    PubMed Central

    Price, Nathan E.; Catalano, Michael J.; Liu, Shuo; Wang, Yinsheng; Gates, Kent S.

    2015-01-01

    A new type of interstrand DNA–DNA cross-link between abasic (Ap) sites and 2′-deoxyadenosine (dA) residues was recently reported, but the chemical structure and properties of this lesion were not rigorously established. Here we characterized the nucleoside cross-link remnant released by enzymatic digestion of duplex DNA containing the dA-Ap cross-link. A synthetic standard was prepared for the putative nucleoside cross-link remnant 6 in which the anomeric carbon of the 2-deoxyribose residue was connected to the exocyclic N6-amino group of dA. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis showed that the synthetic material 6 matched the authentic cross-link remnant released by enzymatic digestion of cross-linked DNA. These findings establish the chemical structure of the dA-Ap cross-link released from duplex DNA and may provide methods for the detection of this lesion in cellular DNA. Both the nucleoside cross-link remnant 6 and the cross-link in duplex DNA were quite stable at pH 7 and 37°C, suggesting that the dA-Ap cross-link could be a persistent lesion with the potential to block the action of various DNA processing enzymes. PMID:25779045

  17. Selective amyloid β oligomer assay based on abasic site-containing molecular beacon and enzyme-free amplification.

    PubMed

    Zhu, Linling; Zhang, Junying; Wang, Fengyang; Wang, Ya; Lu, Linlin; Feng, Chongchong; Xu, Zhiai; Zhang, Wen

    2016-04-15

    Amyloid-beta (Aβ) oligomers are highly toxic species in the process of Aβ aggregation and are regarded as potent therapeutic targets and diagnostic markers for Alzheimer's disease (AD). Herein, a label-free molecular beacon (MB) system integrated with enzyme-free amplification strategy was developed for simple and highly selective assay of Aβ oligomers. The MB system was constructed with abasic site (AP site)-containing stem-loop DNA and a fluorescent ligand 2-amino-5,6,7-trimethyl-1,8-naphyridine (ATMND), of which the fluorescence was quenched upon binding to the AP site in DNA stem. Enzyme-free amplification was realized by target-triggered continuous opening of two delicately designed MBs (MB1 and MB2). Target DNA hybridization with MB1 and then MB2 resulted in the release of two ATMND molecules in one binding event. Subsequent target recycling could greatly amplify the detection sensitivity due to the greatly enhanced turn-on emission of ATMND fluorescence. Combining with Aβ oligomers aptamers, the strategy was applied to analyze Aβ oligomers and the results showed that it could quantify Aβ oligomers with high selectivity and monitor the Aβ aggregation process. This novel method may be conducive to improve the diagnosis and pathogenic study of Alzheimer's disease. PMID:26613510

  18. Processing of abasic site damaged lesions by APE1 enzyme on DNA adsorbed over normal and organomodified clay.

    PubMed

    Kumari, Bhavini; Banerjee, Shib Shankar; Singh, Vandana; Das, Prolay; Bhowmick, Anil K

    2014-10-01

    The efficiency of the apurinic/apyrimidinic endonuclease (APE1) DNA repair enzyme in the processing of abasic site DNA damage lesions at precise location in DNA oligomer duplexes that are adsorbed on clay surfaces was evaluated. Three different forms of clay namely montmorillonite, quaternary ammonium salt modified montmorillonite and its boiled counterpart i.e. partially devoid of organic moiety were used for a comparative study of adsorption, desorption and DNA repair efficiency on their surfaces. The interaction between the DNA and the clay was analysed by X-ray diffraction, Atomic force microscopy, UV-Vis spectroscopy and Infrared spectroscopy. The abasic site cleavage efficiency of APE1 enzyme was quantitatively evaluated by polyacrylamide gel electrophoresis. Apart from the difference in the DNA adsorption or desorption capacity of the various forms of clay, substantial variation in the repair efficiency of abasic sites initiated by the APE1 enzyme on the clay surfaces was observed. The incision efficiency of APE1 enzyme at abasic sites was found to be greatly diminished, when the DNA was adsorbed over organomodified montmorillonite. The reduced repair activity indicates an important role of the pendant surfactant groups on the clay surfaces in directing APE1 mediated cleavage of abasic site DNA damage lesions. PMID:25048946

  19. Synthesis, thermal stability and reactivity towards 9-aminoellipticine of double-stranded oligonucleotides containing a true abasic site.

    PubMed Central

    Bertrand, J R; Vasseur, J J; Rayner, B; Imbach, J L; Paoletti, J; Paoletti, C; Malvy, C

    1989-01-01

    A 13 mers abasic oligonucleotide was synthetized. It was therefore possible to compare thermal stability and reactivity of duplex oligonucleotides either with an apurinic/apyrimidinic site or without any lesion. An important decrease in the melting temperature appeared for duplexes with an abasic site. The chemical reaction of these modified oligonucleotides with the intercalating agent 9-aminoellipticine was studied by gel electrophoresis and by fluorescence. The formation of a Schiff base between 9-aminoellipticine and abasic sites was rapid and complete with duplexes at 11 degrees C. Schiff base related fluorescence and beta-elimination cleavage were more important with the apyrimidinic sites than with the apurinic ones. When compared to previous results obtained with the model d(TprpT) some unexpected behaviours appeared with longer and duplex oligonucleotides. For instance only partial beta-elimination cleavage was observed. It is likely that stacking parameters in the double helix play a great role in the studied reaction. Images PMID:2602153

  20. ALKBH1 is dispensable for abasic site cleavage during base excision repair and class switch recombination.

    PubMed

    Müller, Tina A; Yu, Kefei; Hausinger, Robert P; Meek, Katheryn

    2013-01-01

    Potential roles of the abasic site lyase activity associated with AlkB homolog 1 (ALKBH1) were assessed by studies focusing on the two cellular processes that create abasic sites as intermediates: base excision repair and class switch recombination. Alkbh1(-/-) pups (lacking exon 3) were born at a lower than expected frequency from heterozygous parents, suggesting a reduced survival rate and non-Mendelian inheritance, and they exhibited a gender bias in favor of males (70% males and 30% females). To study ALKBH1's potential involvement in DNA repair, fibroblasts were isolated from Alkbh1(-/-) mice, spontaneously immortalized and tested for resistance to DNA damaging agents. Alkbh1(-/-) and isogenic cells expressing hALKBH1 showed no difference in survival to the DNA damaging agents methyl-methionine sulfate or H2O2. This result indicates that ALKBH1 does not play a major role in the base excision repair pathway. To assess ALKBH1's role in class switch recombination, splenic B cells were isolated from Alkbh1(-/-) and Alkbh1(+/+) mice and subjected to switching from IgM to IgG1. No differences were found in IgG1 switching, suggesting that Alkbh1 is not involved in class switch recombination of the immunoglobulin heavy chain during B lymphocyte activation. PMID:23825659

  1. Correlation of Thermal Stability and Structural Distortion of DNA Interstrand Cross-Links Produced from Oxidized Abasic Sites with Their Selective Formation and Repair.

    PubMed

    Ghosh, Souradyuti; Greenberg, Marc M

    2015-10-13

    C4'-oxidized (C4-AP) and C5'-oxidized abasic sites (DOB) that are produced following abstraction of a hydrogen atom from the DNA backbone reversibly form cross-links selectively with dA opposite a 3'-adjacent nucleotide, despite the comparable proximity of an opposing dA. A previous report on UvrABC incision of DNA substrates containing stabilized analogues of the ICLs derived from C4-AP and DOB also indicated that the latter is repaired more readily by nucleotide excision repair [Ghosh, S., and Greenberg, M. M. (2014) Biochemistry 53, 5958-5965]. The source for selective cross-link formation was probed by comparing the reactivity of ICL analogues of C4-AP and DOB that mimic the preferred and disfavored cross-links with that of reagents that indirectly detect distortion by reacting with the nucleobases. The disfavored C4-AP and DOB analogues were each more reactive than the corresponding preferred cross-link substrates, suggesting that the latter are more stable, which is consistent with selective ICL formation. In addition, the preferred DOB analogue is more reactive than the respective C4-AP ICL, which is consistent with its more efficient incision by UvrABC. The conclusions drawn from the chemical probing experiments are corroborated by UV melting studies. The preferred ICLs exhibit melting temperatures higher than those of the corresponding disfavored isomers. These studies suggest that oxidized abasic sites form reversible interstrand cross-links with dA opposite the 3'-adjacent thymidine because these products are more stable and the thermodynamic preference is reflected in the transition states for their formation. PMID:26426430

  2. Impeded repair of abasic site damaged lesions in DNA adsorbed over functionalized multiwalled carbon nanotube and graphene oxide.

    PubMed

    Kumari, Rina; Mondal, Titash; Bhowmick, Anil K; Das, Prolay

    2016-06-01

    The processing of abasic site DNA damage lesions in extracellular DNA in the presence of engineered carbon nanomaterials (CNMs) is demonstrated. The efficacy of the apurinic-apyrimidinic endonuclease 1 (APE1) in the cleavage of abasic site lesions in the presence of carboxylated multi-walled carbon nanotubes (MWCNT-COOH) and graphene oxide (GO) are compared. The CNMs were found to perturb the incision activity of APE1. The reason for such perturbation process was anticipated to take place either by the non-specific adsorption of APE1 over the free surface of the CNMs or steric hindrance offered by the CNM-DNA complex. Accordingly, bovine serum albumin (BSA) was selectively utilized to block the free surface of the CNM-DNA hybrid material. Further treatment of the CNM-DNA-BSA complex with APE1 resulted in a marginal increase in APE1 efficiency. This indicates that APE1 in solution is unable to process the abasic sites on DNA adsorbed over the CNMs. However, the cleavage activity of APE1 was restored in the presence of non-ionic surfactant (Tween 20) that inhibits adsorption of the DNA on the surface of the CNMs. The conformational deformation of the DNA, along with steric hindrance induced by the CNMs resulted in the inhibition of abasic site DNA repair by APE1. Moreover, appreciable changes in the secondary structure of APE1 adsorbed over the CNMs were observed that contribute further to the repair refractivity of the abasic sites. From a toxicological viewpoint, these findings can be extended to the study of the effect of engineered nanoparticles in the intracellular DNA repair process. PMID:27265379

  3. Covalent Adduct Formation between the Antihypertensive Drug Hydralazine and Abasic Sites in Double- and Single-Stranded DNA

    PubMed Central

    2015-01-01

    Hydralazine (4) is an antihypertensive agent that displays both mutagenic and epigenetic properties. Here, gel electrophoretic, mass spectroscopic, and chemical kinetics methods were used to provide evidence that medicinally relevant concentrations of 4 rapidly form covalent adducts with abasic sites in double- and single-stranded DNA under physiological conditions. These findings raise the intriguing possibility that the genotoxic properties of this clinically used drug arise via reactions with an endogenous DNA lesion rather than with the canonical structure of DNA. PMID:25405892

  4. Recognition of triplex forming oligodeoxynucleotides incorporating abasic sites by 5-arylcytosine residues in duplex DNAs.

    PubMed

    Mizuta, Masahiro; Banba, Jun-Ichi; Kanamori, Takashi; Ohkubo, Akihiro; Sekine, Mitsuo; Seio, Kohji

    2007-01-01

    In this paper, we reported our attempt to use a 5arylcytosine (dC(ar)) and the abasic site () as an artificial base pair for DNA triplex. The idea was confirmed by the molecular modeling studied in which the aromatic group of (ph) which protrudes in the major groove was buried into the cleft formed by the residue in the TFO. We synthesized three kinds of dC(ar) and the oligonucleotides incorporating them. Our UV-melting experiments revealed that the DNA triplex containing the dC(ph).phi was more stable than that containing dC.phi pair. Moreover, the dC.phi pair was more stable than any other dC.Y pairs such as dC(ph).G, dC(ph).C, dC(ph).T and dC(ph).A. These results indicated the possibility that the appropriate pair of dC(Ar) and could be the new sequence code of DNA triplex. We also carried out the Tm analyses of other TFOs incorporating dC(Ar) and , and clarified the stability of these triplexes. PMID:18029568

  5. Evidence for abasic site sugar phosphate-mediated cytotoxicity in alkylating agent treated Saccharomyces cerevisiae.

    PubMed

    Heacock, Michelle; Poltoratsky, Vladimir; Prasad, Rajendra; Wilson, Samuel H

    2012-01-01

    To better understand alkylating agent-induced cytotoxicity and the base lesion DNA repair process in Saccharomyces cerevisiae, we replaced the RAD27(FEN1) open reading frame (ORF) with the ORF of the bifunctional human repair enzyme DNA polymerase (Pol) β. The aim was to probe the effect of removal of the incised abasic site 5'-sugar phosphate group (i.e., 5'-deoxyribose phosphate or 5'-dRP) in protection against methyl methanesulfonate (MMS)-induced cytotoxicity. In S. cerevisiae, Rad27(Fen1) was suggested to protect against MMS-induced cytotoxicity by excising multinucleotide flaps generated during repair. However, we proposed that the repair intermediate with a blocked 5'-end, i.e., 5'-dRP group, is the actual cytotoxic lesion. In providing a 5'-dRP group removal function mediated by dRP lyase activity of Pol β, the effects of the 5'-dRP group were separated from those of the multinucleotide flap itself. Human Pol β was expressed in S. cerevisiae, and this partially rescued the MMS hypersensitivity observed with rad27(fen1)-null cells. To explore this rescue effect, altered forms of Pol β with site-directed eliminations of either the 5'-dRP lyase or polymerase activity were expressed in rad27(fen1)-null cells. The 5'-dRP lyase, but not the polymerase activity, conferred the resistance to MMS. These results suggest that after MMS exposure, the 5'-dRP group in the repair intermediate is cytotoxic and that Rad27(Fen1) protection against MMS in wild-type cells is due to elimination of the 5'-dRP group. PMID:23144716

  6. Accelerated processing of solitary and clustered abasic site DNA damage lesions by APE1 in the presence of aqueous extract of Ganoderma lucidum.

    PubMed

    Kumari, Bhavini; DAS, Prolay; Kumari, Rekha

    2016-06-01

    The stimulatory effect of the aqueous extract of G. lucidum, a basidiomycetes class fungus in the APE1-enzyme-mediated processing of solitary and bistranded clustered abasic sites DNA damages is presented. Abasic sites are considered the most common type of DNA damage lesions. Our study shows enhanced activity of APE1 in the processing of abasic sites in the presence of the polysaccharides fraction of G. lucidum. Remarkable increase in the amount of single-strand breaks (SSBs) and double-strand breaks (DSBs) from solitary and bistranded clustered abasic sites respectively with APE1 in the presence of the extract was found. This trend is maintained when abasic sites in DNA oligomers are exposed to fibroblast cell extracts in the presence of the extract. While DNA conformational alteration is negligible, APE1 enzyme shows characteristic changes in the alpha helix and beta strand ratio after incubation with G. lucidum extract. The enhanced reactivity of APE1 at the molecular level in the presence of G. lucidium is attributed to this effect. This study potentially amplifies the scope of the use of G. lucidum, which was earlier shown to have only reactive oxygen species (ROS) scavenging properties with regards to DNA damage inhibition. PMID:27240987

  7. Target-controlled formation of silver nanoclusters in abasic site-incorporated duplex DNA for label-free fluorescence detection of theophylline

    NASA Astrophysics Data System (ADS)

    Park, Ki Soo; Oh, Seung Soo; Soh, H. Tom; Park, Hyun Gyu

    2014-08-01

    A novel, label-free, fluorescence based sensor for theophylline has been developed. In the new sensor system, an abasic site-incorporated duplex DNA probe serves as both a pocket for recognition of theophylline and a template for the preparation of fluorescent silver nanoclusters. The strategy relies on theophylline-controlled formation of fluorescent silver nanoclusters from abasic site-incorporated duplex DNA. When theophylline is not present, silver ions interact with the cytosine groups opposite to the abasic site in duplex DNA. This interaction leads to efficient formation of intensely red fluorescent silver nanoclusters. In contrast, when theophylline is bound at the abasic site through pseudo base-pairing with appropriately positioned cytosines, silver ion binding to the cytosine nucleobase is prevented. Consequently, fluorescent silver nanoclusters are not formed causing a significant reduction of the fluorescence signal. By employing this new sensor, theophylline can be highly selectively detected at a concentration as low as 1.8 μM. Finally, the diagnostic capability and practical application of this sensor were demonstrated by its use in detecting theophylline in human blood serum.A novel, label-free, fluorescence based sensor for theophylline has been developed. In the new sensor system, an abasic site-incorporated duplex DNA probe serves as both a pocket for recognition of theophylline and a template for the preparation of fluorescent silver nanoclusters. The strategy relies on theophylline-controlled formation of fluorescent silver nanoclusters from abasic site-incorporated duplex DNA. When theophylline is not present, silver ions interact with the cytosine groups opposite to the abasic site in duplex DNA. This interaction leads to efficient formation of intensely red fluorescent silver nanoclusters. In contrast, when theophylline is bound at the abasic site through pseudo base-pairing with appropriately positioned cytosines, silver ion binding to

  8. Target-controlled formation of silver nanoclusters in abasic site-incorporated duplex DNA for label-free fluorescence detection of theophylline.

    PubMed

    Park, Ki Soo; Oh, Seung Soo; Soh, H Tom; Park, Hyun Gyu

    2014-09-01

    A novel, label-free, fluorescence based sensor for theophylline has been developed. In the new sensor system, an abasic site-incorporated duplex DNA probe serves as both a pocket for recognition of theophylline and a template for the preparation of fluorescent silver nanoclusters. The strategy relies on theophylline-controlled formation of fluorescent silver nanoclusters from abasic site-incorporated duplex DNA. When theophylline is not present, silver ions interact with the cytosine groups opposite to the abasic site in duplex DNA. This interaction leads to efficient formation of intensely red fluorescent silver nanoclusters. In contrast, when theophylline is bound at the abasic site through pseudo base-pairing with appropriately positioned cytosines, silver ion binding to the cytosine nucleobase is prevented. Consequently, fluorescent silver nanoclusters are not formed causing a significant reduction of the fluorescence signal. By employing this new sensor, theophylline can be highly selectively detected at a concentration as low as 1.8 μM. Finally, the diagnostic capability and practical application of this sensor were demonstrated by its use in detecting theophylline in human blood serum. PMID:24901073

  9. Interaction of the recombinant human methylpurine-DNA glycosylase (MPG protein) with oligodeoxyribonucleotides containing either hypoxanthine or abasic sites.

    PubMed Central

    Miao, F; Bouziane, M; O'Connor, T R

    1998-01-01

    Methylpurine-DNA glycosylases (MPG proteins, 3-methyladenine-DNA glycosylases) excise numerous damaged bases from DNA during the first step of base excision repair. The damaged bases removed by these proteins include those induced by both alkylating agents and/or oxidizing agents. The intrinsic kinetic parameters (k(cat) and K(m)) for the excision of hypoxanthine by the recombinant human MPG protein from a 39 bp oligodeoxyribonucleotide harboring a unique hypoxanthine were determined. Comparison with other reactions catalyzed by the human MPG protein suggests that the differences in specificity are primarily in product release and not binding. Analysis of MPG protein binding to the 39 bp oligodeoxyribonucleotide revealed that the apparent dissociation constant is of the same order of magnitude as the K(m) and that a 1:1 complex is formed. The MPG protein also forms a strong complex with the product of excision, an abasic site, as well as with a reduced abasic site. DNase I footprinting experiments with the MPG protein on an oligodeoxyribonucleotide with a unique hypoxanthine at a defined position indicate that the protein protects 11 bases on the strand with the hypoxanthine and 12 bases on the complementary strand. Competition experiments with different length, double-stranded, hypoxanthine-containing oligodeoxyribonucleotides show that the footprinted region is relatively small. Despite the small footprint, however, oligodeoxyribonucleotides comprising <15 bp with a hypoxanthine have a 10-fold reduced binding capacity compared with hypoxanthine-containing oligodeoxyribonucleotides >20 bp in length. These results provide a basis for other structural studies of the MPG protein with its targets. PMID:9705516

  10. Resistance to Nucleotide Excision Repair of Bulky Guanine Adducts Opposite Abasic Sites in DNA Duplexes and Relationships between Structure and Function

    PubMed Central

    Liu, Zhi; Ding, Shuang; Kropachev, Konstantin; Lei, Jia; Amin, Shantu; Broyde, Suse; Geacintov, Nicholas E.

    2015-01-01

    The nucleotide excision repair of certain bulky DNA lesions is abrogated in some specific non-canonical DNA base sequence contexts, while the removal of the same lesions by the nucleotide excision repair mechanism is efficient in duplexes in which all base pairs are complementary. Here we show that the nucleotide excision repair activity in human cell extracts is moderate-to-high in the case of two stereoisomeric DNA lesions derived from the pro-carcinogen benzo[a]pyrene (cis- and trans-B[a]P-N2-dG adducts) in a normal DNA duplex. By contrast, the nucleotide excision repair activity is completely abrogated when the canonical cytosine base opposite the B[a]P-dG adducts is replaced by an abasic site in duplex DNA. However, base excision repair of the abasic site persists. In order to understand the structural origins of these striking phenomena, we used NMR and molecular spectroscopy techniques to evaluate the conformational features of 11mer DNA duplexes containing these B[a]P-dG lesions opposite abasic sites. Our results show that in these duplexes containing the clustered lesions, both B[a]P-dG adducts adopt base-displaced intercalated conformations, with the B[a]P aromatic rings intercalated into the DNA helix. To explain the persistence of base excision repair in the face of the opposed bulky B[a]P ring system, molecular modeling results suggest how the APE1 base excision repair endonuclease, that excises abasic lesions, can bind productively even with the trans-B[a]P-dG positioned opposite the abasic site. We hypothesize that the nucleotide excision repair resistance is fostered by local B[a]P residue—DNA base stacking interactions at the abasic sites, that are facilitated by the absence of the cytosine partner base in the complementary strand. More broadly, this study sets the stage for elucidating the interplay between base excision and nucleotide excision repair in processing different types of clustered DNA lesions that are substrates of nucleotide

  11. Simple, High-Yield Syntheses of DNA Duplexes Containing Interstrand DNA-DNA Cross-Links Between an N(4) -Aminocytidine Residue and an Abasic Site.

    PubMed

    Gamboa Varela, Jacqueline; Gates, Kent S

    2016-01-01

    The protocol describes the preparation and purification of interstrand DNA-DNA cross-links derived from the reaction of an N(4) -aminocytidine residue with an abasic site in duplex DNA. The procedures employ inexpensive, commercially available chemicals and enzymes to carry out post-synthetic modification of commercially available oligodeoxynucleotides. The yield of cross-linked duplex is typically better than 90%. If purification is required, the cross-linked duplex can be readily separated from single-stranded DNA starting materials by denaturing gel electrophoresis. The resulting covalent hydrazone-based cross-links are stable under physiologically relevant conditions and may be useful for biophysical studies, structural analyses, DNA repair studies, and materials science applications. © 2016 by John Wiley & Sons, Inc. PMID:27248783

  12. Chemical repair of base lesions, AP-sites, and strand breaks on plasmid DNA in dilute aqueous solution by ascorbic acid

    SciTech Connect

    Hata, Kuniki; Urushibara, Ayumi; Yamashita, Shinichi; Shikazono, Naoya; Yokoya, Akinari; Katsumura, Yosuke

    2013-05-03

    Highlights: •We report a novel mechanism of radiation protection of DNA by chemical activity of ascorbic acid. •The “chemical repair” of DNA damage was revealed using biochemical assay and chemical kinetics analysis. •We found that ascorbic acid significantly repairs precursors of nucleobase lesions and abasic sites. •However, ascorbic acid seldom repairs precursors of DNA-strand breaks. -- Abstract: We quantified the damage yields produced in plasmid DNA by γ-irradiation in the presence of low concentrations (10–100 μM) of ascorbic acid, which is a major antioxidant in living systems, to clarify whether it chemically repairs radiation damage in DNA. The yield of DNA single strand breaks induced by irradiation was analyzed with agarose gel electrophoresis as conformational changes in closed circular plasmids. Base lesions and abasic sites were also observed as additional conformational changes by treating irradiated samples with glycosylase proteins. By comparing the suppression efficiencies to the induction of each DNA lesion, in addition to scavenging of the OH radicals derived from water radiolysis, it was found that ascorbic acid promotes the chemical repair of precursors of AP-sites and base lesions more effectively than those of single strand breaks. We estimated the efficiency of the chemical repair of each lesion using a kinetic model. Approximately 50–60% of base lesions and AP-sites were repaired by 10 μM ascorbic acid, although strand breaks were largely unrepaired by ascorbic acid at low concentrations. The methods in this study will provide a route to understanding the mechanistic aspects of antioxidant activity in living systems.

  13. Influence of DNA torsional rigidity on excision of 7,8-dihydro-8-oxo-2′-deoxyguanosine in the presence of opposing abasic sites by human OGG1 protein

    PubMed Central

    Barone, F.; Dogliotti, E.; Cellai, L.; Giordano, C.; Bjørås, M.; Mazzei, F.

    2003-01-01

    The human protein OGG1 (hOGG1) targets the highly mutagenic base 7,8-dihydro-8-oxo-2′-deoxyguanosine (8-oxodG) and shows a high specificity for the opposite DNA base. Abasic sites can arise in DNA in close opposition to 8-oxodG either during repair of mismatched bases (i.e. 8-oxodG/A mismatches) or, more frequently, as a consequence of ionizing radiation exposure. Bistranded DNA lesions may remain unrepaired and lead to cell death via double-strand break formation. In order to explore the role of damaged-DNA dynamics in recognition/excision by the hOGG1 repair protein, specific oligonucleotides containing an 8-oxodG opposite an abasic site, at different relative distances on the complementary strand, were synthesized. Rotational dynamics were studied by means of fluorescence polarization anisotropy decay experiments and the torsional elastic constant as well as the hydrodynamic radius of the DNA fragments were evaluated. Efficiency of excision of 8-oxodG was tested using purified human glycosylase. A close relation between the twisting flexibility of the DNA fragment and the excision efficiency of the oxidative damage by hOGG1 protein within a cluster was found. PMID:12655006

  14. RNA duplexes with abasic substitutions are potent and allele-selective inhibitors of huntingtin and ataxin-3 expression

    PubMed Central

    Liu, Jing; Pendergraff, Hannah; Narayanannair, K. Jayaprakash; Lackey, Jeremy G.; Kuchimanchi, Satya; Rajeev, Kallanthottathil G.; Manoharan, Muthiah; Hu, Jiaxin; Corey, David R.

    2013-01-01

    Abasic substitutions within DNA or RNA are tools for evaluating the impact of absent nucleobases. Because of the importance of abasic sites in genetic damage, most research has involved DNA. Little information is available on the impact of abasic substitutions within RNA or on RNA interference (RNAi). Here, we examine the effect of abasic substitutions on RNAi and allele-selective gene silencing. Huntington's disease (HD) and Machado Joseph Disease (MJD) are severe neurological disorders that currently have no cure. HD and MJD are caused by an expansion of CAG repeats within one mRNA allele encoding huntingtin (HTT) and ataxin-3 (ATX-3) proteins. Agents that silence mutant HTT or ATX-3 expression would remove the cause of HD or MJD and provide an option for therapeutic development. We describe flexible syntheses for abasic substitutions and show that abasic RNA duplexes allele-selectively inhibit both mutant HTT and mutant ATX-3. Inhibition involves the RNAi protein argonaute 2, even though the abasic substitution disrupts the catalytic cleavage of RNA target by argonaute 2. Several different abasic duplexes achieve potent and selective inhibition, providing a broad platform for subsequent development. These findings introduce abasic substitutions as a tool for tailoring RNA duplexes for gene silencing. PMID:23887934

  15. Critical determinants for substrate recognition and catalysis in the M. tuberculosis class II AP-endonuclease/3'-5' exonuclease III.

    PubMed

    Khanam, Taran; Shukla, Ankita; Rai, Niyati; Ramachandran, Ravishankar

    2015-05-01

    The Mycobacterium tuberculosis AP-endonuclease/3'-5' exodeoxyribonuclease (MtbXthA) is an important player in DNA base excision repair (BER). We demonstrate that the enzyme has robust apurinic/apyrimidinic (AP) endonuclease activity, 3'-5' exonuclease, phosphatase, and phosphodiesterase activities. The enzyme functions as an AP-endonuclease at high ionic environments, while the 3'-5'-exonuclease activity is predominant at low ionic environments. Our molecular modelling and mutational experiments show that E57 and D251 are critical for catalysis. Although nicked DNA and gapped DNA are fair substrates of MtbXthA, the gap-size did not affect the excision activity and furthermore, a substrate with a recessed 3'-end is preferred. To understand the determinants of abasic-site recognition, we examined the possible roles of (i) the base opposite the abasic site, (ii) the abasic ribose ring itself, (iii) local distortions in the AP-site, and (iv) conserved residues located near the active site. Our experiments demonstrate that the first three determinants do not play a role in MtbXthA, and in fact the enzyme exhibits robust endonucleolytic activity against single-stranded AP DNA also. Regarding the fourth determinant, it is known that the catalytic-site of AP endonucleases is surrounded by conserved aromatic residues and intriguingly, the exact residues that are directly involved in abasic site recognition vary with the individual proteins. We therefore, used a combination of mutational analysis, kinetic assays, and structure-based modelling, to identify that Y237, supported by Y137, mediates the formation of the MtbXthA-AP-DNA complex and AP-site incision. PMID:25748880

  16. Characterization of diadenosine tetraphosphate (Ap4A) binding sites in cultured chromaffin cells: evidence for a P2y site.

    PubMed Central

    Pintor, J.; Torres, M.; Castro, E.; Miras-Portugal, M. T.

    1991-01-01

    1. Diadenosine tetraphosphate (Ap4A) a dinucleotide, which is stored in secretory granules, presents two types of high affinity binding sites in chromaffin cells. A Kd value of 8 +/- 0.65 x 10(-11) M and Bmax value of 5420 +/- 450 sites per cell were obtained for the high affinity binding site. A Kd value of 5.6 +/- 0.53 x 10(-9) M and a Bmax value close to 70,000 sites per cell were obtained for the second binding site with high affinity. 2. The diadenosine polyphosphates, Ap3A, Ap4A, Ap5A and Ap6A, displaced [3H]-Ap4A from the two binding sites, the Ki values being 1.0 nM, 0.013 nM, 0.013 nM and 0.013 nM for the very high affinity binding site and 0.5 microM, 0.13 microM, 0.062 microM and 0.75 microM for the second binding site. 3. The ATP analogues displaced [3H]-Ap4A with the potency order of the P2y receptors, adenosine 5'-O-(2 thiodiphosphate) (ADP-beta-S) greater than 5'-adenylyl imidodiphosphate (AMP-PNP) greater than alpha, beta-methylene ATP (alpha, beta-MeATP), in both binding sites. The Ki values were respectively 0.075 nM, 0.2 nM and 0.75 nM for the very high affinity binding site and 0.125 microM, 0.5 microM and 0.9 microM for the second binding site. PMID:1912985

  17. Clustered DNA Lesions Containing 5-Formyluracil and AP Site: Repair via the BER System

    PubMed Central

    Belousova, Ekaterina A.; Vasil'eva, Inna A.; Moor, Nina A.; Zatsepin, Timofey S.; Oretskaya, Tatiana S.; Lavrik, Olga I.

    2013-01-01

    Lesions in the DNA arise under ionizing irradiation conditions or various chemical oxidants as a single damage or as part of a multiply damaged site within 1–2 helical turns (clustered lesion). Here, we explored the repair opportunity of the apurinic/apyrimidinic site (AP site) composed of the clustered lesion with 5-formyluracil (5-foU) by the base excision repair (BER) proteins. We found, that if the AP site is shifted relative to the 5-foU of the opposite strand, it could be repaired primarily via the short-patch BER pathway. In this case, the cleavage efficiency of the AP site-containing DNA strand catalyzed by human apurinic/apyrimidinic endonuclease 1 (hAPE1) decreased under AP site excursion to the 3'-side relative to the lesion in the other DNA strand. DNA synthesis catalyzed by DNA polymerase lambda was more accurate in comparison to the one catalyzed by DNA polymerase beta. If the AP site was located exactly opposite 5-foU it was expected to switch the repair to the long-patch BER pathway. In this situation, human processivity factor hPCNA stimulates the process. PMID:23936307

  18. Methanopyrus kandleri topoisomerase V contains three distinct AP lyase active sites in addition to the topoisomerase active site.

    PubMed

    Rajan, Rakhi; Osterman, Amy; Mondragón, Alfonso

    2016-04-20

    Topoisomerase V (Topo-V) is the only topoisomerase with both topoisomerase and DNA repair activities. The topoisomerase activity is conferred by a small alpha-helical domain, whereas the AP lyase activity is found in a region formed by 12 tandem helix-hairpin-helix ((HhH)2) domains. Although it was known that Topo-V has multiple repair sites, only one had been mapped. Here, we show that Topo-V has three AP lyase sites. The atomic structure and Small Angle X-ray Scattering studies of a 97 kDa fragment spanning the topoisomerase and 10 (HhH)2domains reveal that the (HhH)2domains extend away from the topoisomerase domain. A combination of biochemical and structural observations allow the mapping of the second repair site to the junction of the 9th and 10th (HhH)2domains. The second site is structurally similar to the first one and to the sites found in other AP lyases. The 3rd AP lyase site is located in the 12th (HhH)2domain. The results show that Topo-V is an unusual protein: it is the only known protein with more than one (HhH)2domain, the only known topoisomerase with dual activities and is also unique by having three AP lyase repair sites in the same polypeptide. PMID:26908655

  19. Methanopyrus kandleri topoisomerase V contains three distinct AP lyase active sites in addition to the topoisomerase active site

    PubMed Central

    Rajan, Rakhi; Osterman, Amy; Mondragón, Alfonso

    2016-01-01

    Topoisomerase V (Topo-V) is the only topoisomerase with both topoisomerase and DNA repair activities. The topoisomerase activity is conferred by a small alpha-helical domain, whereas the AP lyase activity is found in a region formed by 12 tandem helix-hairpin-helix ((HhH)2) domains. Although it was known that Topo-V has multiple repair sites, only one had been mapped. Here, we show that Topo-V has three AP lyase sites. The atomic structure and Small Angle X-ray Scattering studies of a 97 kDa fragment spanning the topoisomerase and 10 (HhH)2 domains reveal that the (HhH)2 domains extend away from the topoisomerase domain. A combination of biochemical and structural observations allow the mapping of the second repair site to the junction of the 9th and 10th (HhH)2 domains. The second site is structurally similar to the first one and to the sites found in other AP lyases. The 3rd AP lyase site is located in the 12th (HhH)2 domain. The results show that Topo-V is an unusual protein: it is the only known protein with more than one (HhH)2 domain, the only known topoisomerase with dual activities and is also unique by having three AP lyase repair sites in the same polypeptide. PMID:26908655

  20. Slow base excision by human alkyladenine DNA glycosylase limits the rate of formation of AP sites and AP endonuclease 1 does not stimulate base excision.

    PubMed

    Maher, Robyn L; Vallur, Aarthy C; Feller, Joyce A; Bloom, Linda B

    2007-01-01

    The base excision repair pathway removes damaged DNA bases and resynthesizes DNA to replace the damage. Human alkyladenine DNA glycosylase (AAG) is one of several damage-specific DNA glycosylases that recognizes and excises damaged DNA bases. AAG removes primarily damaged adenine residues. Human AP endonuclease 1 (APE1) recognizes AP sites produced by DNA glycosylases and incises the phophodiester bond 5' to the damaged site. The repair process is completed by a DNA polymerase and DNA ligase. If not tightly coordinated, base excision repair could generate intermediates that are more deleterious to the cell than the initial DNA damage. The kinetics of AAG-catalyzed excision of two damaged bases, hypoxanthine and 1,N6-ethenoadenine, were measured in the presence and absence of APE1 to investigate the mechanism by which the base excision activity of AAG is coordinated with the AP incision activity of APE1. 1,N6-ethenoadenine is excised significantly slower than hypoxanthine and the rate of excision is not affected by APE1. The excision of hypoxanthine is inhibited to a small degree by accumulated product, and APE1 stimulates multiple turnovers by alleviating product inhibition. These results show that APE1 does not significantly affect the kinetics of base excision by AAG. It is likely that slow excision by AAG limits the rate of AP site formation in vivo such that AP sites are not created faster than can be processed by APE1. PMID:17018265

  1. ANKRD1 acts as a transcriptional repressor of MMP13 via the AP-1 site.

    PubMed

    Almodóvar-García, Karinna; Kwon, Minjae; Samaras, Susan E; Davidson, Jeffrey M

    2014-04-01

    The transcriptional cofactor ANKRD1 is sharply induced during wound repair, and its overexpression enhances healing. We recently found that global deletion of murine Ankrd1 impairs wound contraction and enhances necrosis of ischemic wounds. A quantitative PCR array of Ankrd1(-/-) (KO) fibroblasts indicated that ANKRD1 regulates MMP genes. Yeast two-hybrid and coimmunoprecipitation analyses associated ANKRD1 with nucleolin, which represses AP-1 activation of MMP13. Ankrd1 deletion enhanced both basal and phorbol 12-myristate 13-acetate (PMA)-induced MMP13 promoter activity; conversely, Ankrd1 overexpression in control cells decreased PMA-induced MMP13 promoter activity. Ankrd1 reconstitution in KO fibroblasts decreased MMP13 mRNA, while Ankrd1 knockdown increased these levels. MMP13 mRNA and protein were elevated in intact skin and wounds of KO versus Ankrd1(fl/fl) (FLOX) mice. Electrophoretic mobility shift assay gel shift patterns suggested that additional transcription factors bind to the MMP13 AP-1 site in the absence of Ankrd1, and this concept was reinforced by chromatin immunoprecipitation analysis as greater binding of c-Jun to the AP-1 site in extracts from FLOX versus KO fibroblasts. We propose that ANKRD1, in association with factors such as nucleolin, represses MMP13 transcription. Ankrd1 deletion additionally relieved MMP10 transcriptional repression. Nuclear ANKRD1 appears to modulate extracellular matrix remodeling by MMPs. PMID:24515436

  2. ANKRD1 Acts as a Transcriptional Repressor of MMP13 via the AP-1 Site

    PubMed Central

    Almodóvar-García, Karinna; Kwon, Minjae; Samaras, Susan E.

    2014-01-01

    The transcriptional cofactor ANKRD1 is sharply induced during wound repair, and its overexpression enhances healing. We recently found that global deletion of murine Ankrd1 impairs wound contraction and enhances necrosis of ischemic wounds. A quantitative PCR array of Ankrd1−/− (KO) fibroblasts indicated that ANKRD1 regulates MMP genes. Yeast two-hybrid and coimmunoprecipitation analyses associated ANKRD1 with nucleolin, which represses AP-1 activation of MMP13. Ankrd1 deletion enhanced both basal and phorbol 12-myristate 13-acetate (PMA)-induced MMP13 promoter activity; conversely, Ankrd1 overexpression in control cells decreased PMA-induced MMP13 promoter activity. Ankrd1 reconstitution in KO fibroblasts decreased MMP13 mRNA, while Ankrd1 knockdown increased these levels. MMP13 mRNA and protein were elevated in intact skin and wounds of KO versus Ankrd1fl/fl (FLOX) mice. Electrophoretic mobility shift assay gel shift patterns suggested that additional transcription factors bind to the MMP13 AP-1 site in the absence of Ankrd1, and this concept was reinforced by chromatin immunoprecipitation analysis as greater binding of c-Jun to the AP-1 site in extracts from FLOX versus KO fibroblasts. We propose that ANKRD1, in association with factors such as nucleolin, represses MMP13 transcription. Ankrd1 deletion additionally relieved MMP10 transcriptional repression. Nuclear ANKRD1 appears to modulate extracellular matrix remodeling by MMPs. PMID:24515436

  3. Rescue of abasic hammerhead ribozymes by exogenous addition of specific bases.

    PubMed

    Peracchi, A; Beigelman, L; Usman, N; Herschlag, D

    1996-10-15

    We have synthesized 13 hammerhead ribozyme variants, each containing an abasic residue at a specific position of the catalytic core. The activity of each of the variants is significantly reduced. In four cases, however, activity can be rescued by exogenous addition of the missing base. For one variant, the rescue is 300-fold; for another, the rescue is to the wild-type level. This latter abasic variant (G10.1X) has been characterized in detail. Activation is specific for guanine, the base initially removed. In addition, the specificity for guanine versus adenine is substantially altered by replacing C with U in the opposite strand of the ribozyme. These results show that a binding site for a small, noncharged ligand can be created in a preexisting ribozyme structure. This has implications for structure-function analysis of RNA, and leads to speculations about evolution in an "RNA world" and about the potential therapeutic use of ribozymes. PMID:8876168

  4. Regulation of human alcohol dehydrogenase gene ADH7: importance of an AP-1 site.

    PubMed

    Kotagiri, S; Edenberg, H J

    1998-07-01

    The structure and function of the human alcohol dehydrogenase 7 (ADH7) promoter were analyzed. A promoter fragment extending to bp -232 functioned well in H4IIE-C3, CV-1, and HeLa cells, whereas the region extending further upstream to bp -799 had no significant effect on activity. We identified cis-acting elements in the proximal 232 bp and examined their effect on promoter activity. Mutation of site A, where c-Jun bound, caused a drastic decrease in the promoter activity in H4IIE-C3 and CV-1 cells, suggesting that AP-1 plays an important role in the regulation of ADH7. Mutation of site B also caused a large drop in promoter activity in both cell lines; C/EBPalpha can bind to this site, but because the site affects activity approximately equally in CV-1 cells that lack C/EBPalpha and in H4IIE-C3 cells that contain low levels, other proteins are likely to play the major roles in vivo. Mutation of site C, where C/EBP bound and c-Jun bound weakly, had different effects in the two cell lines: in H4IIE-C3 cells, the site C mutation did not significantly increase promoter activity, whereas in CV-1 cells, which lack C/EBPalpha, it led to a doubling of activity. Surprisingly, cotransfection of the wild-type promoter with C/EBPa or C/EBPbeta led to a decrease in promoter activity, which might in part explain the lack of activity of ADH7 in adult liver. PMID:9703017

  5. Chemical repair activity of free radical scavenger edaravone: reduction reactions with dGMP hydroxyl radical adducts and suppression of base lesions and AP sites on irradiated plasmid DNA

    PubMed Central

    Hata, Kuniki; Urushibara, Ayumi; Yamashita, Shinichi; Lin, Mingzhang; Muroya, Yusa; Shikazono, Naoya; Yokoya, Akinari; Fu, Haiying; Katsumura, Yosuke

    2015-01-01

    Reactions of edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one) with deoxyguanosine monophosphate (dGMP) hydroxyl radical adducts were investigated by pulse radiolysis technique. Edaravone was found to reduce the dGMP hydroxyl radical adducts through electron transfer reactions. The rate constants of the reactions were greater than 4 × 108 dm3 mol−1 s−1 and similar to those of the reactions of ascorbic acid, which is a representative antioxidant. Yields of single-strand breaks, base lesions, and abasic sites produced in pUC18 plasmid DNA by gamma ray irradiation in the presence of low concentrations (10–1000 μmol dm−3) of edaravone were also quantified, and the chemical repair activity of edaravone was estimated by a method recently developed by the authors. By comparing suppression efficiencies to the induction of each DNA lesion, it was found that base lesions and abasic sites were suppressed by the chemical repair activity of edaravone, although the suppression of single-strand breaks was not very effective. This phenomenon was attributed to the chemical repair activity of edaravone toward base lesions and abasic sites. However, the chemical repair activity of edaravone for base lesions was lower than that of ascorbic acid. PMID:25212600

  6. Arsenite suppression of involucrin transcription through AP1 promoter sites in cultured human keratinocytes

    SciTech Connect

    Sinitsyna, Nadezda N.; Reznikova, Tatiana V.; Qin Qin; Song, Hyukhwan; Phillips, Marjorie A.; Rice, Robert H.

    2010-03-15

    While preserving keratinocyte proliferative ability, arsenite suppresses cellular differentiation markers by preventing utilization of AP1 transcriptional response elements. In present experiments, arsenite had a dramatic effect in electrophoretic mobility supershift analysis of proteins binding to an involucrin promoter AP1 response element. Without arsenite treatment, binding of JunB and Fra1 was readily detected in nuclear extracts from preconfluent cultures and was not detected a week after confluence, while c-Fos was detected only after confluence. By contrast, band shift of nuclear extracts from arsenite treated cultures showed only JunB and Fra1 binding in postconfluent as well as preconfluent cultures. Immunoblotting of cell extracts showed that arsenite treatment prevented the loss of Fra1 and the increase in c-Fos proteins that occurred after confluence in untreated cultures. Chromatin immunoprecipitation assays demonstrated substantial reduction of c-Fos and acetylated histone H3 at the proximal and distal AP1 response elements in the involucrin promoter and of coactivator p300 at the proximal element. Alteration of AP1 transcription factors was also examined in response to treatment with four metal containing compounds (chromate, vanadate, hemin, divalent cadmium) that also suppress involucrin transcription. These agents all influenced transcription at AP1 elements in a transcriptional reporter assay, but exhibited less effect than arsenite on binding activity assessed by mobility shift and chromatin immunoprecipitation and displayed variable effects on AP1 protein levels. These findings help trace a mechanism by which transcriptional effects of arsenite become manifest and help rationalize the unique action of arsenite, compared to the other agents, to preserve proliferative ability.

  7. Negative regulation of the rat stromelysin gene promoter by retinoic acid is mediated by an AP1 binding site.

    PubMed Central

    Nicholson, R C; Mader, S; Nagpal, S; Leid, M; Rochette-Egly, C; Chambon, P

    1990-01-01

    Stromelysin is a member of the metalloproteinase family which plays an important role in extracellular matrix remodelling during many normal and disease processes. We show here that in polyomavirus-transformed rat embryo fibroblast cells (PyT21), the transcription from the stromelysin gene is repressed by the vitamin A derivative retinoic acid (RA). Furthermore, expression vectors encoding the human RA receptors hRAR-alpha, hRAR-beta and hRAR-gamma repress chloramphenicol acetyltransferase (CAT) expression from stromelysin promoter-CAT gene expression vectors in RA-treated PyT21 and human HeLa cells, as determined by transient transfection assays. Through mutation and deletion analysis, we show that the RA dependent repression is mediated by a 25 bp region from nucleotide positions -72 to -48 of the rat stromelysin 5'-flanking DNA sequence. Further mutation analysis of this region indicates that the DNA sequence required for RA dependent repression colocalizes with an AP1 binding site which is essential for promoter activity. We show also that RA represses the transcriptional activity of a reporter gene containing a TPA responding AP1 binding site driving the HSV tk promoter. Thus the RAR-RA complex appears to repress transcription of the stromelysin gene by blocking activation by positive regulatory factors. However, we found no evidence supporting the possibility that the RA dependent repression could be due to RAR binding to the AP1 binding site or to the AP1 components c-fos and c-jun. Images Fig. 1. Fig. 2. Fig. 4. Fig. 6. Fig. 7. Fig. 8. PMID:2176152

  8. Disruption of adaptor protein 2μ (AP-2μ) in cochlear hair cells impairs vesicle reloading of synaptic release sites and hearing.

    PubMed

    Jung, SangYong; Maritzen, Tanja; Wichmann, Carolin; Jing, Zhizi; Neef, Andreas; Revelo, Natalia H; Al-Moyed, Hanan; Meese, Sandra; Wojcik, Sonja M; Panou, Iliana; Bulut, Haydar; Schu, Peter; Ficner, Ralf; Reisinger, Ellen; Rizzoli, Silvio O; Neef, Jakob; Strenzke, Nicola; Haucke, Volker; Moser, Tobias

    2015-11-01

    Active zones (AZs) of inner hair cells (IHCs) indefatigably release hundreds of vesicles per second, requiring each release site to reload vesicles at tens per second. Here, we report that the endocytic adaptor protein 2μ (AP-2μ) is required for release site replenishment and hearing. We show that hair cell-specific disruption of AP-2μ slows IHC exocytosis immediately after fusion of the readily releasable pool of vesicles, despite normal abundance of membrane-proximal vesicles and intact endocytic membrane retrieval. Sound-driven postsynaptic spiking was reduced in a use-dependent manner, and the altered interspike interval statistics suggested a slowed reloading of release sites. Sustained strong stimulation led to accumulation of endosome-like vacuoles, fewer clathrin-coated endocytic intermediates, and vesicle depletion of the membrane-distal synaptic ribbon in AP-2μ-deficient IHCs, indicating a further role of AP-2μ in clathrin-dependent vesicle reformation on a timescale of many seconds. Finally, we show that AP-2 sorts its IHC-cargo otoferlin. We propose that binding of AP-2 to otoferlin facilitates replenishment of release sites, for example, via speeding AZ clearance of exocytosed material, in addition to a role of AP-2 in synaptic vesicle reformation. PMID:26446278

  9. 3CAPS – a structural AP–site analogue as a tool to investigate DNA base excision repair

    PubMed Central

    Schuermann, David; Scheidegger, Simon P.; Weber, Alain R.; Bjørås, Magnar; Leumann, Christian J.; Schär, Primo

    2016-01-01

    Abasic sites (AP-sites) are frequent DNA lesions, arising by spontaneous base hydrolysis or as intermediates of base excision repair (BER). The hemiacetal at the anomeric centre renders them chemically reactive, which presents a challenge to biochemical and structural investigation. Chemically more stable AP-site analogues have been used to avoid spontaneous decay, but these do not fully recapitulate the features of natural AP–sites. With its 3′–phosphate replaced by methylene, the abasic site analogue 3CAPS was suggested to circumvent some of these limitations. Here, we evaluated the properties of 3CAPS in biochemical BER assays with mammalian proteins. 3CAPS-containing DNA substrates were processed by APE1, albeit with comparably poor efficiency. APE1-cleaved 3CAPS can be extended by DNA polymerase β but repaired only by strand displacement as the 5′–deoxyribophosphate (dRP) cannot be removed. DNA glycosylases physically and functionally interact with 3CAPS substrates, underlining its structural integrity and biochemical reactivity. The AP lyase activity of bifunctional DNA glycosylases (NTH1, NEIL1, FPG), however, was fully inhibited. Notably, 3CAPS-containing DNA also effectively inhibited the activity of bifunctional glycosylases on authentic substrates. Hence, the chemically stable 3CAPS with its preserved hemiacetal functionality is a potent tool for BER research and a potential inhibitor of bifunctional DNA glycosylases. PMID:26733580

  10. Regulation of the HMOX1 gene by the transcription factor AP-2δ with unique DNA binding site.

    PubMed

    Sun, Liyun; Zhao, Yuxia; Gu, Shaohua; Mao, Yumin; Ji, Chaoneng; Xin, Xiujuan

    2014-07-01

    AP-2 transcription factors are important sequence-specific DNA-binding regulators that are expressed in the neural crest and other tissues during mammalian development. The human AP-2 family of transcription factors consists of five members, AP-2α, -β, -γ, -δ and -ε, which have an important role in the regulation of gene expression during development and in the differentiation of multiple organs and tissues. The present study aimed to investigate the mechanism by which AP-2δ mediates heme oxygenase-1 (HMOX1) gene expression. It was identified that the human AP-2δ protein exhibited weak binding to a suboptimal AP-2 sequence, 5'-GCCN3GGC-3', to which all other AP-2 proteins bind in vitro, providing the first example of DNA target specificity amongst the AP-2 family. AP-2δ protein bound to an optimized AP-2 consensus DNA sequence, 5'-GCCTGAGGC-3', in vitro and transactivated gene expression in eukaryotic cells. The transactivation domain of Ap-2δ differs notably from those in the other AP-2 proteins as it lacks the PY motif (XPPXY) and several other conserved residues that are important for the transcriptional activity of AP-2 proteins, yet it functions as an equally strong activator. PMID:24789576

  11. Genome-wide analysis of p63 binding sites identifies AP-2 factors as co-regulators of epidermal differentiation

    PubMed Central

    McDade, Simon S.; Henry, Alexandra E.; Pivato, Geraldine P.; Kozarewa, Iwanka; Mitsopoulos, Constantinos; Fenwick, Kerry; Assiotis, Ioannis; Hakas, Jarle; Zvelebil, Marketa; Orr, Nicholas; Lord, Christopher J.; Patel, Daksha; Ashworth, Alan; McCance, Dennis J.

    2012-01-01

    The p63 transcription factor (TP63) is critical in development, growth and differentiation of stratifying epithelia. This is highlighted by the severity of congenital abnormalities caused by TP63 mutations in humans, the dramatic phenotypes in knockout mice and de-regulation of TP63 expression in neoplasia altering the tumour suppressive roles of the TP53 family. In order to define the normal role played by TP63 and provide the basis for better understanding how this network is perturbed in disease, we used chromatin immunoprecipitation combined with massively parallel sequencing (ChIP-seq) to identify >7500 high-confidence TP63-binding regions across the entire genome, in primary human neonatal foreskin keratinocytes (HFKs). Using integrative strategies, we demonstrate that only a subset of these sites are bound by TP53 in response to DNA damage. We identify a role for TP63 in transcriptional regulation of multiple genes genetically linked to cleft palate and identify AP-2alpha (TFAP2A) as a co-regulator of a subset of these genes. We further demonstrate that AP-2gamma (TFAP2C) can bind a subset of these regions and that acute depletion of either TFAP2A or TFAP2C alone is sufficient to reduce terminal differentiation of organotypic epidermal skin equivalents, indicating overlapping physiological functions with TP63. PMID:22573176

  12. Abasic pivot substitution harnesses target specificity of RNA interference

    PubMed Central

    Lee, Hye-Sook; Seok, Heeyoung; Lee, Dong Ha; Ham, Juyoung; Lee, Wooje; Youm, Emilia Moonkyung; Yoo, Jin Seon; Lee, Yong-Seung; Jang, Eun-Sook; Chi, Sung Wook

    2015-01-01

    Gene silencing via RNA interference inadvertently represses hundreds of off-target transcripts. Because small interfering RNAs (siRNAs) can function as microRNAs, avoiding miRNA-like off-target repression is a major challenge. Functional miRNA–target interactions are known to pre-require transitional nucleation, base pairs from position 2 to the pivot (position 6). Here, by substituting nucleotide in pivot with abasic spacers, which prevent base pairing and alleviate steric hindrance, we eliminate miRNA-like off-target repression while preserving on-target activity at ∼80–100%. Specifically, miR-124 containing dSpacer pivot substitution (6pi) loses seed-mediated transcriptome-wide target interactions, repression activity and biological function, whereas other conventional modifications are ineffective. Application of 6pi allows PCSK9 siRNA to efficiently lower plasma cholesterol concentration in vivo, and abolish potentially deleterious off-target phenotypes. The smallest spacer, C3, also shows the same improvement in target specificity. Abasic pivot substitution serves as a general means to harness the specificity of siRNA experiments and therapeutic applications. PMID:26679372

  13. The E6AP Binding Pocket of the HPV16 E6 Oncoprotein Provides a Docking Site for a Small Inhibitory Peptide Unrelated to E6AP, Indicating Druggability of E6

    PubMed Central

    Kintscher, Susanne; Reinz, Eileen; Sehr, Peter; Bulkescher, Julia; Hoppe-Seyler, Karin; Travé, Gilles; Hoppe-Seyler, Felix

    2014-01-01

    The HPV E6 oncoprotein maintains the malignant phenotype of HPV-positive cancer cells and represents an attractive therapeutic target. E6 forms a complex with the cellular E6AP ubiquitin ligase, ultimately leading to p53 degradation. The recently elucidated x-ray structure of a HPV16 E6/E6AP complex showed that HPV16 E6 forms a distinct binding pocket for E6AP. This discovery raises the question whether the E6AP binding pocket is druggable, i. e. whether it provides a docking site for functional E6 inhibitors. To address these issues, we performed a detailed analysis of the HPV16 E6 interactions with two small peptides: (i) E6APpep, corresponding to the E6 binding domain of E6AP, and (ii) pep11**, a peptide that binds to HPV16 E6 and, in contrast to E6APpep, induces apoptosis, specifically in HPV16-positive cancer cells. Surface plasmon resonance, NMR chemical shift perturbation, and mammalian two-hybrid analyses coupled to mutagenesis indicate that E6APpep contacts HPV16 E6 amino acid residues within the E6AP pocket, both in vitro and intracellularly. Many of these amino acids were also important for binding to pep11**, suggesting that the binding sites for the two peptides on HPV16 E6 overlap. Yet, few E6 amino acids were differentially involved which may contribute to the higher binding affinity of pep11**. Data from the HPV16 E6/pep11** interaction allowed the rational design of single amino acid exchanges in HPV18 and HPV31 E6 that enabled their binding to pep11**. Taken together, these results suggest that E6 molecular surfaces mediating E6APpep binding can also accommodate pro-apoptotic peptides that belong to different sequence families. As proof of concept, this study provides the first experimental evidence that the E6AP binding pocket is druggable, opening new possibilities for rational, structure-based drug design. PMID:25383876

  14. Tiron Inhibits UVB-Induced AP-1 Binding Sites Transcriptional Activation on MMP-1 and MMP-3 Promoters by MAPK Signaling Pathway in Human Dermal Fibroblasts

    PubMed Central

    Zhang, Chao; Zhao, Mei; Zhang, Quan-Wu; Gao, Feng-Hou

    2016-01-01

    Recent research found that Tiron was an effective antioxidant that could act as the intracellular reactive oxygen species (ROS) scavenger or alleviate the acute toxic metal overload in vivo. In this study, we investigated the inhibitory effect of Tiron on matrix metalloproteinase (MMP)-1 and MMP-3 expression in human dermal fibroblast cells. Western blot and ELISA analysis revealed that Tiron inhibited ultraviolet B (UVB)-induced protein expression of MMP-1 and MMP-3. Real-time quantitative PCR confirmed that Tiron could inhibit UVB-induced mRNA expression of MMP-1 and MMP-3. Furthermore, Tiron significantly blocked UVB-induced activation of the MAPK signaling pathway and activator protein (AP)-1 in the downstream of this transduction pathway in fibroblasts. Through the AP-1 binding site mutation, it was found that Tiron could inhibit AP-1-induced upregulation of MMP-1 and MMP-3 expression through blocking AP-1 binding to the AP-1 binding sites in the MMP-1 and MMP-3 promoter region. In conclusion, Tiron may be a novel antioxidant for preventing and treating skin photoaging UV-induced. PMID:27486852

  15. Biochemical reconstitution of abasic DNA lesion replication in Xenopus extracts

    PubMed Central

    Liao, Shuren; Matsumoto, Yoshihiro; Yan, Hong

    2007-01-01

    Cellular DNA is under constant attack from numerous exogenous and endogenous agents. The resulting DNA lesions, if not repaired timely, could stall DNA replication, leading to genome instability. To better understand the mechanism of DNA lesion replication at the biochemical level, we have attempted to reconstitute this process in Xenopus egg extracts, the only eukaryotic in vitro system that relies solely on cellular proteins for DNA replication. By using a plasmid DNA that carries a site-specific apurinic/apyrimidinic (AP) lesion as template, we have found that DNA replication is stalled one nucleotide before the lesion. The stalling is temporary and the lesion is eventually replicated by both an error-prone mechanism and an error-free mechanism. This is the first biochemical system that recapitulates efficiently and faithfully all major aspects of DNA lesion replication. It has provided the first direct evidence for the existence of an error-free lesion replication mechanism and also demonstrated that the error-prone mechanism is a major contributor to lesion replication. PMID:17702761

  16. An AP-1 binding site in the enhancer/core element of the HIV-1 promoter controls the ability of HIV-1 to establish latent infection.

    PubMed

    Duverger, Alexandra; Wolschendorf, Frank; Zhang, Mingce; Wagner, Fredric; Hatcher, Brandon; Jones, Jennifer; Cron, Randall Q; van der Sluis, Renee M; Jeeninga, Rienk E; Berkhout, Ben; Kutsch, Olaf

    2013-02-01

    Following integration, HIV-1 in most cases produces active infection events; however, in some rare instances, latent infection events are established. The latter have major clinical implications, as latent infection allows the virus to persist despite antiretroviral therapy. Both the cellular factors and the viral elements that potentially determine whether HIV-1 establishes active or latent infection events remain largely elusive. We detail here the contribution of different long terminal repeat (LTR) sequences for the establishment of latent HIV-1 infection. Using a panel of full-length replication-competent virus constructs that reflect naturally occurring differences of HIV-1 subtype-specific LTRs and targeted LTR mutants, we found the primary ability of HIV-1 to establish latent infection in this system to be controlled by a four-nucleotide (nt) AP-1 element just upstream of the NF-κB element in the viral promoter. Deletion of this AP-1 site mostly deprived HIV-1 of the ability to establish latent HIV-1 infection. Extension of this site to a 7-nt AP-1 sequence massively promoted latency establishment, suggesting that this promoter region represents a latency establishment element (LEE). Given that these minimal changes in a transcription factor binding site affect latency establishment to such large extent, our data support the notion that HIV-1 latency is a transcription factor restriction phenomenon. PMID:23236059

  17. An AP-1 Binding Site in the Enhancer/Core Element of the HIV-1 Promoter Controls the Ability of HIV-1 To Establish Latent Infection

    PubMed Central

    Duverger, Alexandra; Wolschendorf, Frank; Zhang, Mingce; Wagner, Fredric; Hatcher, Brandon; Jones, Jennifer; Cron, Randall Q.; van der Sluis, Renee M.; Jeeninga, Rienk E.; Berkhout, Ben

    2013-01-01

    Following integration, HIV-1 in most cases produces active infection events; however, in some rare instances, latent infection events are established. The latter have major clinical implications, as latent infection allows the virus to persist despite antiretroviral therapy. Both the cellular factors and the viral elements that potentially determine whether HIV-1 establishes active or latent infection events remain largely elusive. We detail here the contribution of different long terminal repeat (LTR) sequences for the establishment of latent HIV-1 infection. Using a panel of full-length replication-competent virus constructs that reflect naturally occurring differences of HIV-1 subtype-specific LTRs and targeted LTR mutants, we found the primary ability of HIV-1 to establish latent infection in this system to be controlled by a four-nucleotide (nt) AP-1 element just upstream of the NF-κB element in the viral promoter. Deletion of this AP-1 site mostly deprived HIV-1 of the ability to establish latent HIV-1 infection. Extension of this site to a 7-nt AP-1 sequence massively promoted latency establishment, suggesting that this promoter region represents a latency establishment element (LEE). Given that these minimal changes in a transcription factor binding site affect latency establishment to such large extent, our data support the notion that HIV-1 latency is a transcription factor restriction phenomenon. PMID:23236059

  18. Efficient cleavage of single and clustered AP site lesions within mono-nucleosome templates by CHO-K1 nuclear extract contrasts with retardation of incision by purified APE1

    PubMed Central

    Eccles, Laura J.; Menoni, Hervé; Angelov, Dimitar; Lomax, Martine E.; O’Neill, Peter

    2015-01-01

    Clustered DNA damage is a unique characteristic of radiation-induced DNA damage and the formation of these sites poses a serious challenge to the cell’s repair machinery. Within a cell DNA is compacted, with nucleosomes being the first order of higher level structure. However, few data are reported on the efficiency of clustered-lesion processing within nucleosomal DNA templates. Here, we show retardation of cleavage of a single AP site by purified APE1 when contained in nucleosomal DNA, compared to cleavage of an AP site in non-nucleosomal DNA. This retardation seen in nucleosomal DNA was alleviated by incubation with CHO-K1 nuclear extract. When clustered DNA damage sites containing bistranded AP sites were present in nucleosomal DNA, efficient cleavage of the AP sites was observed after treatment with nuclear extract. The resultant DSB formation led to DNA dissociating from the histone core and nucleosomal dispersion. Clustered damaged sites containing bistranded AP site/8-oxoG residues showed no retardation of cleavage of the AP site but retardation of 8-oxoG excision, compared to isolated lesions, thus DSB formation was not seen. An increased understanding of processing of clustered DNA damage in a nucleosomal environment may lead to new strategies to enhance the cytotoxic effects of radiotherapeutics. PMID:26439176

  19. Sp1 binds two sites in the CD11c promoter in vivo specifically in myeloid cells and cooperates with AP1 to activate transcription.

    PubMed Central

    Noti, J D; Reinemann, B C; Petrus, M N

    1996-01-01

    The leukocyte integrin gene, CD11c, is transcriptionally regulated and is expressed predominantly on differentiated cells of the myelomonocytic lineage. In this study we have demonstrated that the regions -72 to -63 and -132 to -104 of the CD11c promoter contain elements responsible for phorbol ester-induced differentiation of the myeloid cell line HL60. DNase I footprinting analysis revealed that these regions can bind purified Sp1, and supershift analysis with Sp1 antibody confirmed that Sp1 in HL60 nuclear extracts could bind these regions. Transfection analysis of CD11c promoter-chloramphenicol acetyltransferase constructs containing deletions of these Sp1-binding sites revealed that these sites are essential for expression of the CD11c gene in HL60 cells but not in the T-cell line Molt4 or the cervical carcinoma cell line HeLa. Moreover, cotransfection of pPacSp1 along with these CD11c promoter-chloramphenicol acetyltransferase constructs into Sp1-deficient Drosophila Schneider 2 cells verified that these sites are essential for Sp1-dependent expression of the CD11c promoter. In vivo genomic footprinting revealed that Sp1 contacts the CD11c promoter within the regions -69 to -63 and -116 to -105 in phorbol 12-myristate 13-acetate-differentiated HL60 cells but not in undifferentiated HL60 cells or in Molt4 or HeLa cells. Cotransfection assays in HL60 cells revealed that Sp1 acts synergistically with Ap1 to activate CD11c. Further, both Sp1 sites are capable of cooperating with AP1. In vitro DNase I footprinting analysis with purified Sp1 and c-jun proteins showed that Sp1 binding could facilitate binding of c-jun. We propose that myeloid-specific expression of the CD11c promoter and is facilitated by cooperative interaction between the Sp1- and Ap1-binding sites. PMID:8649405

  20. Elk1 and AP-1 sites in the TBP promoter mediate alcohol-induced deregulation of Pol III-dependent genes

    PubMed Central

    Zhong, Qian; Shi, Ganggang; Zhang, Yanmei; Levy, Daniel; Zhong, Shuping

    2013-01-01

    The major risk factors for hepatocellular carcinoma (HCC) are chronic liver diseases that include hepatitis B, hepatitis C, alcoholic liver disease and non-alcoholic steatohepatitis. However, the mechanisms of alcohol-associated HCC remain to be elucidated. The products of RNA Pol III (RNA polymerase III) dependent genes are elevated in both transformation cells and tumor cells. TBP (TATA-box binding protein) is a central transcription factor, which regulates Pol I, Pol II and Pol III gene activity. Our studies have demonstrated that alcohol increases TBP expression and Pol III gene transcription to promote liver tumor formation. We continue to investigate how ethanol mediates TBP expression. Here, we report that ethanol induces TBP promoter activity and the induction is ethanol dose dependent. Blocking the JNK1 pathway by a chemical inhibitor and siRNA reduce this ethanol-induced activity. Furthermore, mutating G>A at a −46bp Elk1 binding site of the TBP promoter or mutating AP-1 binding site at −37bp (A>G) and −38bp (C>T) reduces the TBP promoter activity. Mutation of both Elk1 and AP-1 binding sites dramatically represses this induction. Together, these studies demonstrate that, for the first time, alcohol increases Pol III gene transcription through a response element, which is composed of the overlapping the Elk1 and AP-1 binding sites of the TBP promoter. It suggests that these binding sites may play a critical role in alcohol-induced deregulation of Pol III genes in liver tumor development. PMID:23454483

  1. The Synonymous Ala87 Mutation of Estrogen Receptor Alpha Modifies Transcriptional Activation Through Both ERE and AP1 Sites.

    PubMed

    Fernández-Calero, Tamara; Flouriot, Gilles; Marín, Mónica

    2016-01-01

    Estrogen receptor α (ERα) exerts regulatory actions through genomic mechanisms. In the classical pathway, ligand-activated ERα binds directly to DNA through estrogen response elements (ERE) located in the promoter of target genes. ERα can also exert indirect regulation of transcription via protein-protein interaction with other transcription factors such as AP-1.S everal ERα synonymous polymorphisms have been identified and efforts to understand their implications have been made. Nevertheless effects of synonymous polymorphisms are still neglected. This chapter focuses on the experimental procedure employed in order to characterize the transcriptional activity of a synonymous polymorphism of the ERα (rs746432) called Alanine 87 (Ala87). Activity of both WT and Ala87 ERα isoforms on transcriptional pathways can be analyzed in transiently transfected cells using different reporter constructs. ERα efficiency on the classical genomic pathway can be analyzed by determining its transactivation activity on an ERE-driven thymidine kinase (TK) promoter controlling the expression of the luciferase reporter gene. Transcriptional activity through the indirect genomic pathway can be analyzed by employing an AP-1 DNA response element-driven promoter also controlling the expression of luciferase reporter gene. PMID:26585143

  2. The regulation of hepcidin expression by serum treatment: requirements of the BMP response element and STAT- and AP-1-binding sites.

    PubMed

    Kanamori, Yohei; Murakami, Masaru; Matsui, Tohru; Funaba, Masayuki

    2014-11-10

    Expression of hepcidin, a central regulator of systemic iron metabolism, is transcriptionally regulated by the bone morphogenetic protein (BMP) pathway. However, the factors other than the BMP pathway also participate in the regulation of hepcidin expression. In the present study, we show that serum treatment increased hepcidin expression and transcription without inducing the phosphorylation of Smad1/5/8 in primary hepatocytes, HepG2 cells or Hepa1-6 cells. Co-treatment with LDN-193189, an inhibitor of the BMP type I receptor, abrogated this hepcidin induction. Reporter assays using mutated reporters revealed the involvement of the BMP response element-1 (BMP-RE1) and signal transducers and activator of transcription (STAT)- and activator protein (AP)-1-binding sites in serum-induced hepcidin transcription in HepG2 cells. Serum treatment induced the expression of the AP-1 components c-fos and junB in primary hepatocytes and HepG2 cells. Forced expression of c-fos or junB enhanced the response of hepcidin transcription to serum treatment. By contrast, the expression of dominant negative (dn)-c-fos and dn-junB decreased hepcidin transcription. The present study reveals that serum contains factors stimulating hepcidin transcription. Basal BMP activity is essential for the serum-induced hepcidin transcription, although serum treatment does not stimulate the BMP pathway. The induction of c-fos and junB by serum treatment stimulates hepcidin transcription, through possibly cooperation with BMP-mediated signaling. Considering that AP-1 is induced by various stimuli, the present results suggest that hepcidin expression is regulated by more diverse factors than had been previously considered. PMID:25151311

  3. Identification of a functional transcriptional factor AP-1 site in the sheep interferon tau gene that mediates a response to PMA in JEG3 cells.

    PubMed Central

    Yamaguchi, H; Ikeda, Y; Moreno, J I; Katsumura, M; Miyazawa, T; Takahashi, E; Imakawa, K; Sakai, S; Christenson, R K

    1999-01-01

    To examine regulatory mechanisms of sheep interferon tau (oIFNtau) gene expression, potential enhancer/silencer elements of the oIFNtau gene were examined using a transient transfection system with oIFNtau gene-chloramphenicol acetyltransferase (oIFNtau-CAT) reporter constructs in human choriocarcinoma cells, JEG3. Experiments with 5'-deletion constructs revealed that the upstream regions from bases -654 to -607 and from bases -606 to -555 were essential for oIFNtau gene expression. In a heterologous transcriptional system in which the upstream regions of oIFNtau were inserted in front of simian virus 40 (SV40) promoter, the regions between bases -654 and -555 were determined as being the enhancer region required for oIFNtau-SV40-CAT transactivation. A subsequent study with the oIFNtau-CAT constructs lacking the upstream region between bases -542 and -124 revealed that, in addition to the further upstream region between bases -1000 and -654, the sequences from bases -543 to -452 seemed to act as silencer regions. The oIFNtau-CAT constructs with site-specific mutagenesis revealed that multiple enhancer elements existed between bases -654 and -555 of the oIFNtau gene. On the basis of nucleotide sequence analysis, there are numerous sites between bases -654 and -555 to which potential transcriptional factors, AP-1, GATA and GATA-related proteins, could bind. Furthermore, gel mobility-shift assays revealed that AP-1 or other nuclear factors could bind to these elements. In co-transfection studies, the expression of c-Jun plus c-Fos enhanced the transactivation of oIFNtau-CAT but the expression of GATA-1, GATA-2 or GATA-3 did not. Taken together, these results suggest that the upstream region between bases -654 and -555 could be considered as the enhancer region for oIFNtau gene transactivation. PMID:10359663

  4. Investigation of the Role of the Histidine-Aspartate Pair in the Human Exonuclease III-like Abasic Endonuclease, Ape1

    SciTech Connect

    Lowry, David F. ); Hoyt, David W. ); Khazi, Fayaz A.; Bagu, John R. ); Lindsey, Andrea G.; Wilson, David M.

    2003-05-30

    Hydrogen bonded histidine-aspartate (His-Asp) pairs are critical constituents in several key enzymatic reactions. To date, the role that these pairs play in catalysis is best understood in serine and trypsin-like proteases, where structural and biochemical NMR studies have revealed important pKa values and hydrogen-bonding patterns within the catalytic pocket. However, the role of the His-Asp pair in metal-assisted catalysis is less clear. Here, we apply liquid state NMR to investigate the role of a critical histidine of apurinic endonuclease 1 (Ape1), a human DNA repair enzyme that cleaves adjacent to abasic sites in DNA using one or more divalent cations and an active site His-Asp pair. The studies within suggest that the Ape1 His- Asp pair functions as neither a general base catalyst nor a metal ligand. Rather, the pair likely stabilizes the pentavalent transition state necessary for phospho-transfer.

  5. Site-directed mutagenesis of the human DNA repair enzyme HAP1: identification of residues important for AP endonuclease and RNase H activity.

    PubMed

    Barzilay, G; Walker, L J; Robson, C N; Hickson, I D

    1995-05-11

    HAP1 protein, the major apurinic/apyrimidinic (AP) endonuclease in human cells, is a member of a homologous family of multifunctional DNA repair enzymes including the Escherichia coli exonuclease III and Drosophila Rrp1 proteins. The most extensively characterised member of this family, exonuclease III, exhibits both DNA- and RNA-specific nuclease activities. Here, we show that the RNase H activity characteristic of exonuclease III has been conserved in the human homologue, although the products resulting from RNA cleavage are dissimilar. To identify residues important for enzymatic activity, five mutant HAP1 proteins containing single amino acid substitutions were purified and analysed in vitro. The substitutions were made at sites of conserved amino acids and targeted either acidic or histidine residues because of their known participation in the active sites of hydrolytic nucleases. One of the mutant proteins (replacement of Asp-219 by alanine) showed a markedly reduced enzymatic activity, consistent with a greatly diminished capacity to bind DNA and RNA. In contrast, replacement of Asp-90, Asp-308 or Glu-96 by alanine led to a reduction in enzymatic activity without significantly compromising nucleic acid binding. Replacement of His-255 by alanine led to only a very small reduction in enzymatic activity. Our data are consistent with the presence of a single catalytic active site for the DNA- and RNA-specific nuclease activities of the HAP1 protein. PMID:7784208

  6. Luminescent platinum(II) complexes with functionalized N-heterocyclic carbene or diphosphine selectively probe mismatched and abasic DNA

    PubMed Central

    Fung, Sin Ki; Zou, Taotao; Cao, Bei; Chen, Tianfeng; To, Wai-Pong; Yang, Chen; Lok, Chun-Nam; Che, Chi-Ming

    2016-01-01

    The selective targeting of mismatched DNA overexpressed in cancer cells is an appealing strategy in designing cancer diagnosis and therapy protocols. Few luminescent probes that specifically detect intracellular mismatched DNA have been reported. Here we used Pt(II) complexes with luminescence sensitive to subtle changes in the local environment and report several Pt(II) complexes that selectively bind to and identify DNA mismatches. We evaluated the complexes' DNA-binding characteristics by ultraviolet/visible absorption titration, isothermal titration calorimetry, nuclear magnetic resonance and quantum mechanics/molecular mechanics calculations. These Pt(II) complexes show up to 15-fold higher emission intensities upon binding to mismatched DNA over matched DNA and can be utilized for both detecting DNA abasic sites and identifying cancer cells and human tissue samples with different levels of mismatch repair. Our work highlights the potential of luminescent Pt(II) complexes to differentiate between normal cells and cancer cells which generally possess more aberrant DNA structures. PMID:26883164

  7. Endonuclease IV cleaves apurinic/apyrimidinic sites in single-stranded DNA and its application for biosensing.

    PubMed

    Kong, Xiang-Juan; Wu, Shuang; Cen, Yao; Chen, Ting-Ting; Yu, Ru-Qin; Chu, Xia

    2016-07-21

    Endonuclease IV (Endo IV), as a DNA repairing enzyme, plays a crucial role in repairing damaged DNA comprising abasic sites to maintain genomic integrity. The cleaving capability of Endo IV to apurinic/apyrimidinic sites (AP) in single-stranded DNA (ssDNA) was demonstrated. It was found that Endo IV has considerably high cleaving activity to AP sites in ssDNA compared with that in double-stranded DNA (dsDNA). The unique feature of Endo IV in cleaving AP sites in ssDNA was further applied to construct a novel dual signal amplified sensing system for highly sensitive enzyme and protein detection by a combination of exonuclease III (Exo III)-aided cyclic amplification reaction and a rolling circle replication (RCR) technique, which showed a good sensing performance with a detection limit of 0.008 U mL(-1) for Endo IV and 2.5 pM for streptavidin. In addition, the developed method had considerably high specificity for Endo IV and streptavidin over other potential interferences. The developed strategy indeed provides a novel platform for protein and enzyme assays and may find a broad spectrum of applications in bioanalysis, disease diagnosis, and drug development. PMID:27186607

  8. Mfsd2a-based pharmacological strategies for drug delivery across the blood-brain barrier.

    PubMed

    Wang, Jing-Zhang; Xiao, Ning; Zhang, Ying-Zhou; Zhao, Chao-Xian; Guo, Xin-Hua; Lu, Li-Min

    2016-02-01

    The blood-brain barrier (BBB) keeps the central nervous system (CNS) safe from various brain diseases, while the BBB makes it difficult for effective drugs to enter the CNS. Mfsd2a is specifically expressed on the cell membrane of brain-microvascular endothelial cell (BMEC) and is implicated in the delivery of some substances across the BBB. Mfsd2a is the first inhibitor of the transcytosis and the first transporter for lysophosphatidylcholine-docosahexaenoic acid (LPC-DHA) in BMECs. The crucial dual function of Mfsd2a puts forward two kinds of Mfsd2a-based strategies for carrying drugs from blood to the CNS. First, the reversible inhibition of Mfsd2a may temporarily induce a general disinhibition of the transcytosis in BMECs to transport macromolecular drugs across the BBB (Strategy One). Second, Mfsd2a could be used for the transport of some small-molecule drugs chemically coupled to LPC across the BBB (Strategy Two), which is quite similar to the carrier-mediated transport (CMT) via the glucose transporter (GluT1) and the L-type amino acid transporter 1 (LAT1). We here analyze and discuss the clinical significance of the two Mfsd2a-based strategies, including therapeutic potential, available pharmaceuticals, side effects, administration procedures, and disease types. In summary, the regulatory role of Mfsd2a deepens our knowledge of the function of the BBB, potentially contributing to the effective drug delivery in the treatments for neurodegenerative diseases, brain tumors, and life-threatening infections in the CNS. PMID:26747400

  9. Induction of Abasic Sites by the Drinking-Water Mutagen MX in Salmonella TA100

    EPA Science Inventory

    Mutagen X (MX) is a chlorinated furanone that accounts for more of the mutagenic activity of drinking water than any other disinfection by-product. It is one of the most potent base-substitution mutagens in the Salmonella (Ames) mutagenicity assay, producing primarily GC to TA mu...

  10. An Estrogen Receptor-α/p300 Complex Activates the BRCA-1 Promoter at an AP-1 Site That Binds Jun/Fos Transcription Factors: Repressive Effects of p53 on BRCA-1 Transcription1

    PubMed Central

    Jeffy, Brandon D; Hockings, Jennifer K; Kemp, Michael Q; Morgan, Sherif S; Hager, Jill A; Beliakoff, Jason; Whitesell, Luke J; Bowden, G. Timothy; Romagnolo, Donato F

    2005-01-01

    Abstract One of the puzzles in cancer predisposition is that women carrying BRCA-1 mutations preferentially develop tumors in epithelial tissues of the breast and ovary. Moreover, sporadic breast tumors contain lower levels of BRCA-1 in the absence of mutations in the BRCA-1 gene. The problem of tissue specificity requires analysis of factors that are unique to tissues of the breast. For example, the expression of estrogen receptor-α (ERα) is inversely correlated with breast cancer risk, and 90% of BRCA-1 tumors are negative for ERα. Here, we show that estrogen stimulates BRCA-1 promoter activity in transfected cells and the recruitment of ERα and its cofactor p300 to an AP-1 site that binds Jun/Fos transcription factors. The recruitment of ERα/p300 coincides with accumulation in the S-phase of the cell cycle and is antagonized by the antiestrogen tamoxifen. Conversely, we document that overexpression of wild-type p53 prevents the recruitment of ERα to the AP-1 site and represses BRCA-1 promoter activity. Taken together, our findings support a model in which an ERα/AP-1 complex modulates BRCA-1 transcription under conditions of estrogen stimulation. Conversely, the formation of this transcription complex is abrogated in cells overexpressing p53. PMID:16229810

  11. APS Education and Diversity Efforts

    NASA Astrophysics Data System (ADS)

    Prestridge, Katherine; Hodapp, Theodore

    2015-11-01

    American Physical Society (APS) has a wide range of education and diversity programs and activities, including programs that improve physics education, increase diversity, provide outreach to the public, and impact public policy. We present the latest programs spearheaded by the Committee on the Status of Women in Physics (CSWP), with highlights from other diversity and education efforts. The CSWP is working to increase the fraction of women in physics, understand and implement solutions for gender-specific issues, enhance professional development opportunities for women in physics, and remedy issues that impact gender inequality in physics. The Conferences for Undergraduate Women in Physics, Professional Skills Development Workshops, and our new Professional Skills program for students and postdocs are all working towards meeting these goals. The CSWP also has site visit and conversation visit programs, where department chairs request that the APS assess the climate for women in their departments or facilitate climate discussions. APS also has two significant programs to increase participation by underrepresented minorities (URM). The newest program, the APS National Mentoring Community, is working to provide mentoring to URM undergraduates, and the APS Bridge Program is an established effort that is dramatically increasing the number of URM PhDs in physics.

  12. Detecting single-abasic residues within a DNA strand immobilized in a biological nanopore using an integrated CMOS sensor

    PubMed Central

    Kim, Jungsuk; Maitra, Raj D.; Pedrotti, Ken; Dunbar, William B.

    2013-01-01

    In this paper, we demonstrate the application of a novel current-measuring sensor (CMS) customized for nanopore applications. The low-noise CMS is fabricated in a 0.35μm CMOS process and is implemented in experiments involving DNA captured in an α-hemolysin (α-HL) nanopore. Specifically, the CMS is used to build a current amplitude map as a function of varying positions of a single-abasic residue within a homopolymer cytosine single-stranded DNA (ssDNA) that is captured and held in the pore. Each ssDNA is immobilized using a biotin-streptavidin linkage. Five different DNA templates are measured and compared: one all-cytosine ssDNA, and four with a single-abasic residue substitution that resides in or near the ~1.5nm aperture of the α-HL channel when the strand is immobilized. The CMOS CMS is shown to resolves the ~5Å displacements of the abasic residue within the varying templates. The demonstration represents an advance in application-specific circuitry that is optimized for small-footprint nanopore applications, including genomic sequencing. PMID:24496266

  13. AP-Gate

    ERIC Educational Resources Information Center

    Whitman, Glenn

    2003-01-01

    In May 2001, students in the author's Advanced Placement (AP) United States History class were embroiled in a controversy surrounding the AP exam, in particular, having access to the exam's Document Based Question (DBQ) and free response portion prior to the test's administration. Prior to the exam, the College Board had provided a fifty-year time…

  14. A functional activating protein 1 (AP-1) site regulates matrix metalloproteinase 2 (MMP-2) transcription by cardiac cells through interactions with JunB-Fra1 and JunB-FosB heterodimers.

    PubMed Central

    Bergman, Marina R; Cheng, Sunfa; Honbo, Norman; Piacentini, Lucia; Karliner, Joel S; Lovett, David H

    2003-01-01

    Enhanced synthesis of a specific matrix metalloproteinase, MMP-2, has been demonstrated in experimental models of ventricular failure and in cardiac extracts from patients with ischaemic cardiomyopathy. Cultured neonatal rat cardiac fibroblasts and myocytes were used to analyse the determinants of MMP-2 synthesis, including the effects of hypoxia. Culture of rat cardiac fibroblasts for 24 h in 1% oxygen enhanced MMP-2 synthesis by more than 5-fold and augmented the MMP-2 synthetic responses of these cells to endothelin-1, angiotensin II and interleukin 1beta. A series of MMP-2 promoter-luciferase constructs were used to map the specific enhancer element(s) that drive MMP-2 transcription in cardiac cells. Deletion studies mapped a region of potent transactivating function within the 91 bp region from -1433 to -1342 bp, the activity of which was increased by hypoxia. Oligonucleotides from this region were cloned in front of a heterologous simian-virus-40 (SV40) promoter and mapped the enhancer activity to a region between -1410 and -1362 bp that included a potential activating protein 1 (AP-1)-binding sequence, C(-1394)CTGACCTCC. Site-specific mutagenesis of the core TGAC sequence (indicated in bold) eliminated the transactivating activity within the -1410 to -1362 bp sequence. Electrophoretic mobility shift assays (EMSAs) using the -1410 to -1362 bp oligonucleotide and rat cardiac fibroblast nuclear extracts demonstrated specific nuclear-protein binding that was eliminated by cold competitor oligonucleotide, but not by the AP-1-mutated oligonucleotide. Antibody-supershift EMSAs of nuclear extracts from normoxic rat cardiac fibroblasts demonstrated Fra1 and JunB binding to the -1410 to -1362 bp oligonucleotide. Nuclear extracts isolated from hypoxic rat cardiac fibroblasts contained Fra1, JunB and also included FosB. Co-transfection of cardiac fibroblasts with Fra1-JunB and FosB-JunB expression plasmids led to significant increases in transcriptional activity. These

  15. Intrinsically photosensitive retinal ganglion cells detect light with a vitamin A-based photopigment, melanopsin

    PubMed Central

    Fu, Yingbin; Zhong, Haining; Wang, Min-Hua H.; Luo, Dong-Gen; Liao, Hsi-Wen; Maeda, Hidetaka; Hattar, Samer; Frishman, Laura J.; Yau, King-Wai

    2005-01-01

    In mammals, intrinsically photosensitive retinal ganglion cells (ipRGCs) mediate non-image-forming visual functions such as pupillary light reflex (PLR) and circadian photoentrainment. This photosensitivity requires melanopsin, an invertebrate opsin-like protein expressed by the ipRGCs. The precise role of melanopsin remains uncertain. One suggestion has been that melanopsin may be a photoisomerase, serving to regenerate an unidentified pigment in ipRGCs. This possibility was echoed by a recent report that melanopsin is expressed also in the mouse retinal pigment epithelium (RPE), a key center for regeneration of rod and cone pigments. To address this question, we studied mice lacking RPE65, a protein essential for the regeneration of rod and cone pigments. Rpe65-/- ipRGCs were ≈20- to 40-fold less photosensitive than normal at both single-cell and behavioral (PLR) levels but were rescued by exogenous 9-cis-retinal (an 11-cis-retinal analog), indicating the requirement of a vitamin A-based chromophore for ipRGC photosensitivity. In contrast, 9-cis-retinal was unable to restore intrinsic photosensitivity to melanopsin-ablated ipRGCs, arguing against melanopsin functioning merely in photopigment regeneration. Interestingly, exogenous all-trans-retinal was also able to rescue the low sensitivity of rpe65-/- ipRGCs, suggesting that melanopsin could be a bistable pigment. Finally, we detected no melanopsin in the RPE and no changes in rod and cone sensitivities due to melanopsin ablation. Together, these results strongly suggest that melanopsin is the photopigment in the ipRGCs. PMID:16014418

  16. Lipid A-based affinity biosensor for screening anti-sepsis components from herbs.

    PubMed

    Yao, Jie; Chen, Yiguo; Wang, Ning; Jiang, Dongneng; Zheng, Jiang

    2014-01-01

    LPS (lipopolysaccharide), an outer membrane component of Gram-negative bacteria, plays an important role in the pathogenesis of sepsis and lipid A is known to be essential for its toxicity. Therefore it could be an effective measure to prevent sepsis by neutralizing or destroying LPS. Numerous studies have indicated that many traditional Chinese medicines are natural antagonists of LPS in vitro and in vivo. The goal of this study is to develop a rapid method to screen anti-sepsis components from Chinese herbs by use of a direct lipid A-based affinity biosensor technology based on a resonant mirror. The detergent OG (n-octyl β-D-glucopyranoside) was immobilized on a planar non-derivatized cuvette which provided an alternative surface to bind the terminal hydrophilic group of lipid A. A total of 78 herbs were screened based on the affinity biosensor with a target of lipid A. The aqueous extract of PSA (Paeonia suffruticosa Andr) was found to possess the highest capability of binding lipid A. Therefore an aqueous extraction from this plant was investigated further by our affinity biosensor, polyamide chromatography and IEC-HPLC. Finally, we obtained a component (PSA-I-3) from Paeonia suffruticosa Andr that was evaluated with the affinity biosensor. We also studied the biological activities of PSA-I-3 against sepsis in vitro and in vivo to further confirm the component we screened with the biosensor. In vitro, we found that PSA-I-3 could decrease TNFα (tumour necrosis factor α) release from RAW264.7 cells induced by LPS in a dose-dependent manner. In vivo, it increased remarkably the survival of KM (KunMing) mice by challenging both lethal-dose LPS and heat-killed Escherichia coli compared with control groups. Our results suggest that the constructed affinity biosensor can successfully screen the anti-sepsis component from Chinese herbs. PMID:24654965

  17. APS Science 2006.

    SciTech Connect

    Gibson, J. M.; Fenner, R. B.; Long, G.; Borland, M.; Decker, G.

    2007-05-24

    In my five years as the Director of the Advanced Photon Source (APS), I have been fortunate to see major growth in the scientific impact from the APS. This year I am particularly enthusiastic about prospects for our longer-term future. Every scientific instrument must remain at the cutting edge to flourish. Our plans for the next generation of APS--an APS upgrade--got seriously in gear this year with strong encouragement from our users and sponsors. The most promising avenue that has emerged is the energy-recovery linac (ERL) (see article on page xx), for which we are beginning serious R&D. The ERL{at}APS would offer revolutionary performance, especially for x-ray imaging and ultrafast science, while not seriously disrupting the existing user base. I am very proud of our accelerator physics and engineering staff, who not only keep the current APS at the forefront, but were able to greatly impress our international Machine Advisory Committee with the quality of their work on the possible upgrade option (see page xx). As we prepare for long-term major upgrades, our plans to develop and optimize all the sectors at APS in the near future are advancing. Several new beamlines saw first light this year, including a dedicated powder diffraction beamline (11-BM), two instruments for inelastic x-ray scattering at sector 30, and the Center for Nanoscale Materials (CNM) Nanoprobe beamline at sector 26. Our partnership in the first x-ray free-electron laser (LCLS) to be built at Stanford contributes to revolutionary growth in ultrafast science (see page xx), and we are developing a pulse chirping scheme to get ps pulses at sector 7 of the APS within a year or so. In this report, you will find selected highlights of scientific research at the APS from calendar year 2006. The highlighted work covers diverse disciplines, from fundamental to applied science. In the article on page xx you can see the direct impact of APS research on technology. Several new products have emerged from

  18. AP1- and NF-kappaB-binding sites conserved among mammalian WNT10B orthologs elucidate the TNFalpha-WNT10B signaling loop implicated in carcinogenesis and adipogenesis.

    PubMed

    Katoh, Masuko; Katoh, Masaru

    2007-04-01

    WNT signals are context-dependently transduced to canonical and non-canonical signaling cascades. We cloned and characterized wild-type human WNT10B, while another group cloned aberrant human WNT10B with Gly60Asp amino-acid substitution. Proto-oncogene WNT10B is expressed in gastric cancer, pancreatic cancer, breast cancer, esophageal cancer, and cervical cancer. Because WNT10B blocks adipocyte differentiation, coding SNP of WNT10B gene is associated with familial obesity. In 2001, we reported WNT10B upregulation by TNFalpha. Here, comparative integromics analyses on WNT10B orthologs were performed to elucidate the transcriptional mechanism of WNT10B. Chimpanzee WNT10B and cow Wnt10b genes were identified within NW_001223159.1 and AC150975.2 genome sequences, respectively, by using bioinformatics (Techint) and human intelligence (Humint). Chimpanzee WNT10B and cow Wnt10b showed 98.7% and 95.1% total-amino-acid identity with human WNT10B, respectively. N-terminal signal peptide, 24 Cys residues, two Asn-linked glycosylation sites, and Gly60 of human WNT10B were conserved among mammalian WNT10B orthologs. Transcription start site of human WNT10B gene was 106-bp upstream of NM_003394.2 RefSeq 5'-end. Number of GC di-nucleotide repeats just down-stream of WNT10B transcription start site varied among primates and human population. Comparative genomics analyses revealed that double AP1-binding sites in the 5'-flanking promoter region and NF-kappaB-binding site in intron 3 were conserved among human, chimpanzee, cow, mouse, and rat WNT10B orthologs. Because TNFalpha signaling through TNFR1 and TRADD/RIP/TRAF2 complex activates JUN kinase (JNK) and IkappaB kinase (IKK) signaling cascades, conserved AP1- and NF-kappaB-binding sites explain the mechanism of TNFalpha-induced WNT10B upregulation. TNFalpha-WNT10B signaling loop is the negative feedback mechanism of adipogenesis to prevent obesity and metabolic syndrome. On the other hand, TNFalpha-WNT10B signaling loop is

  19. APS Science 2009.

    SciTech Connect

    Gibson, J. M; Mills, D. M.; Gerig, R.

    2010-05-01

    It is my pleasure to introduce the 2009 annual report of the Advanced Photon Source. This was a very good year for us. We operated with high reliability and availability, despite growing problems with obsolete systems, and our users produced a record output of publications. The number of user experiments increased by 14% from 2008 to more than 3600. We congratulate the recipients of the 2009 Nobel Prize in Chemistry-Venkatraman Ramakrishnan (Cambridge Institute for Medical Research), Thomas Steitz (Yale University), and Ada Yonath (Weizmann Institute) - who did a substantial amount of this work at APS beamlines. Thanks to the efforts of our users and staff, and the ongoing counsel of the APS Scientific Advisory Committee, we made major progress in advancing our planning for the upgrade of the APS (APS-U), producing a proposal that was positively reviewed. We hope to get formal approval in 2010 to begin the upgrade. With advocacy from our users and the support of our sponsor, the Office of Basic Energy Sciences in the Department of Energy (DOE) Office of Science, our operating budgets have grown to the level needed to more adequately staff our beamlines. We were also extremely fortunate to have received $7.9 M in American Recovery and Reinvestment Act ('stimulus') funding to acquire new detectors and improve several of our beamlines. The success of the new Linac Coherent Light Source at Stanford, the world's first x-ray free-electron laser, made us particularly proud since the undulators were designed and built by the APS. Among other highlights, we note that more than one-quarter of the 46 Energy Frontier Research Centers, funded competitively across the U.S. in 2009 by the DOE, included the Advanced Photon Source in their proposed work, which shows that synchrotron radiation, and the APS in particular, are central to energy research. While APS research covers everything from fundamental to applied science (reflected by the highlights in this report), the challenge

  20. Development of the AP Technology Through Time

    NASA Astrophysics Data System (ADS)

    Charmier, F.; Martin, O.; Gariepy, R.

    2015-02-01

    This article presents the historical development of the AP Technology (Aluval, Voreppe, France) pot series starting with the AP13 in the 1960s, followed by the AP18 and the AP30 in the 1980s and 1990s. For most of the modern-era technology, from the late 1970s on, development has been based on a three-stage pattern, the first one being the pot modeling, followed by the pot prototype stage, and then the industrial stage, which fully validated the technology. This development pattern has proven to be successful since it has led to the very robust AP Technology pot design generation and a large number of greenfield smelters built in the 1990-2010 selected the AP Technology design. AP60 is the latest of this series: The development at the prototype level was initiated in the 1990s and is presented in the article. It is now followed by the first industrial realization at Jonquière with the startup in late 2013 and the full validation of the technology in mid-2014. The development of APXe, which aims at very low energy consumption, uses many common elements pertaining to the AP60 design and is presented in the article. AP Technology has also addressed the need for continuous and fast improvement of pot performances adapted to each existing client or site specifics; for this purpose, a new development methodology has recently emerged thanks to the very high modeling capabilities. This methodology, based on the validation of "technology bricks" and their integration in the final design following a strict process, is presented in the last section of this article.

  1. Design and synthesis of simple, yet potent and selective non-ring-A pyripyropene A-based inhibitors of acyl-coenzyme A: cholesterol acyltransferase 2 (ACAT2).

    PubMed

    Zhan, Yang; Zhang, Xiao-Wei; Xiong, Ying; Li, Bo-Liang; Nan, Fa-Jun

    2016-01-14

    A series of pyripyropene A-based compounds were designed and synthesized by opening the upper section of the A-ring, which significantly simplifies the structure and synthesis from commercially available starting materials. Representative compound (-)-3 exhibited potent activity against ACAT2 and greater selectivity for ACAT2 than for ACAT1. PMID:26584338

  2. Advancing beyond AP Courses

    ERIC Educational Resources Information Center

    Hammond, Bruce G.

    2009-01-01

    A quiet revolution is picking up steam in the nation's private secondary schools, with broad implications for college admissions and for teaching and learning on both sides of the transition from high school to college. About 50 of the nation's leading college-preparatory schools have opted out of the College Board's Advanced Placement (AP)…

  3. APS power supply controls

    SciTech Connect

    Saunders, C.W.; Despe, O.D.

    1994-03-31

    The purpose of this document is to provide comprehensive coverage of the APS power supply control design. This includes application software, embedded controller software, networks, and hardware. The basic components will be introduced first, followed by the requirements driving the overall design. Subsequent sections will address each component of the design one by one. Latter sections will address specific applications.

  4. The AP Descriptive Chemistry Question: Student Errors

    ERIC Educational Resources Information Center

    Crippen, Kent; Brooks, David W.

    2005-01-01

    For over a decade, the authors have been involved in a design theory experiment providing software for high school students preparing for the descriptive question on the Advanced Placement (AP) chemistry examination. Since 1997, the software has been available as a Web site offering repeatable practice. This study describes a 4-year project during…

  5. Gapminder: An AP Human Geography Lab Assignment

    ERIC Educational Resources Information Center

    Keller, Kenneth H.

    2012-01-01

    This lesson is designed as a lab assignment for Advanced Placement (AP) Human Geography students wherein they use the popular Gapminder web site to compare levels of development in countries from different world regions. For this lesson, it is important for the teacher to practice with Gapminder before giving the assignment to students. (Contains…

  6. Nucleolin binds specifically to an AP-1 DNA sequence and represses AP1-dependent transactivation of the matrix metalloproteinase-13 gene.

    PubMed

    Samuel, Shaija; Twizere, Jean-Claude; Beifuss, Katherine K; Bernstein, Lori R

    2008-01-01

    Transcriptional regulation via activator protein-1 (AP-1) protein binding to AP-1 binding sites within gene promoter regions of AP-1 target genes plays a key role in controlling cellular invasion, proliferation, and oncogenesis, and is important to pathogenesis of arthritis and cardiovascular disease. To identify new proteins that interact with the AP-1 DNA binding site, we performed the DNA affinity chromatography-based Nucleotide Affinity Preincubation Specificity TEst of Recognition (NAPSTER) assay, and discovered a 97 kDa protein that binds in vitro to a minimal AP-1 DNA sequence element. Mass spectrometric fragmentation sequencing determined that p97 is nucleolin. Immunoblotting of DNA affinity-purified material with anti-nucleolin antibodies confirmed this identification. Nucleolin also binds the AP-1 site in gel shift assays. Nucleolin interacts in NAPSTER with the AP-1 site within the promoter sequence of the metalloproteinase-13 gene (MMP-13), and binds in vivo in chromatin immunoprecipitation assays in the vicinity of the AP-1 site in the MMP-13 promoter. Overexpression of nucleolin in human HeLa cervical carcinoma cells significantly represses AP-1 dependent gene transactivation of a minimal AP-1 reporter construct and of an MMP-13 promoter reporter sequence. This is the first report of nucleolin binding and transregulation at the AP-1 site. PMID:17626252

  7. Ku antigen displays the AP lyase activity on a certain type of duplex DNA.

    PubMed

    Kosova, Anastasiya A; Khodyreva, Svetlana N; Lavrik, Olga I

    2016-09-01

    In the search for proteins reactive to apurinic/apyrimidinic (AP) sites, it has been earlier found that proteins of human cell extracts formed the Schiff-base-dependent covalent adduct with an apparent molecular mass of 100kDa with a partial DNA duplex containing an AP site and 5'- and 3'-protruding ends (DDE-AP DNA). The adduct of such electrophoretic mobility was characteristic of only DDE-AP DNA (Ilina et al., Biochem. Biophys. Acta 1784 (2008) 1777-1785). The protein in this unusual adduct was identified as the Ku80 subunit of Ku antigen by peptide mass mapping based on MALDI-TOF MS data (Kosova et al., Biopolym. Cell 30 (2014) 42-46). Here we studied the interaction of Ku with DDE-AP DNA in details. Purified Ku (the Ku80 subunit) was shown to form the 100-kDa adduct highly specific for AP DNA with a certain length of protruding ends, base opposite the AP site and AP site location. Ku is capable of AP site cleavage in DDE-AP DNA unlike in analogous AP DNA with blunt ends. Ku cleaves AP sites via β-elimination and prefers apurinic sites over apyrimidinic ones. The AP site in DDE-DNA can be repaired in an apurinic/apyrimidinic endonuclease-independent manner via the successive action of Ku (cleavage of the AP site), tyrosyl-DNA phosphodiesterase 1 (removal of the 3'-deoxyribose residue), polynucleotide kinase 3'-phosphatase (removal of the 3'-phosphate), DNA polymerase β (incorporation of dNMP), and DNA ligase (sealing the nick). These results provide a new insight into the role of Ku in the repair of AP sites. PMID:27129632

  8. APS Science 2007.

    SciTech Connect

    Not Available

    2008-05-30

    This report provides research highlights from the Advanced Photon Source (APS). Although these highlights represent less than 10% of the published work from the APS in 2007, they give a flavor of the diversity and impact of user research at the facility. In the strategic planning the aim is to foster the growth of existing user communities and foresee new areas of research. This coming year finds the APS engaged in putting together, along with the users, a blueprint for the next five years, and making the case for a set of prioritized investments in beamlines, the accelerator, and infrastructure, each of which will be transformational in terms of scientific impact. As this is written plans are being formulated for an important user workshop on October 20-21, 2008, to prioritize strategic plans. The fruit from past investments can be seen in this report. Examples include the creation of a dedicated beamline for x-ray photon correlation spectroscopy at Sector 8, the evolution of dedicated high-energy x-ray scattering beamlines at sectors 1 and 11, a dedicated imaging beamline at Sector 32, and new beamlines for inelastic scattering and powder diffraction. A single-pulse facility has been built in collaboration with Sector 14 (BioCARS) and Phil Anfinrud at the National Institutes of Health, which will offer exceptionally high flux for single-pulse diffraction. The nanoprobe at Sector 26, built and operated jointly by the Argonne Center for Nanoscale Materials and the X-ray Operations and Research (XOR) section of the APS X-ray Science Division, has come on line to define the state of the art in nanoscience.

  9. Substituting CF2 for O4' in Components of Nucleic Acids: Towards Systems with Reduced Propensity to Form Abasic Lesions.

    PubMed

    Yurenko, Yevgen P; Novotný, Jan; Sklenář, Vladimir; Marek, Radek

    2015-12-01

    Intrinsic structural features and energetics of nucleotides containing variously fluorinated sugars as potential building blocks of DNA duplexes and quadruplexes are explored systematically using the modern methods of density functional theory (DFT) and quantum chemical topology (QCT). Our results suggest that fluorination at the 2'-β or 2'-α,β positions somewhat stabilizes in vacuo the AI relative to the BI conformations. In contrast, substitution of the CF2 group for the O4' atom (O4'-CF2 modification) leads to a preference of the BI relative to AI DNA-like conformers. All the studied modifications result in a noticeable increase in the stability of the glycosidic bond [estimated by the relaxed force constants (RFC) approach], with particularly encouraging results for the O4'-CF2 derivative. Consequently, the O4'-CF2 modified systems are suggested and explored as promising scaffolds for the development of duplex and quadruplex structures with reduced propensity to form abasic lesions and to undergo DNA damage. PMID:26493955

  10. AP-42 REVISION: COKE OVENS

    EPA Science Inventory

    The document "Compilation of Air Pollutant Emission Factors" (AP-42) has been published by the U. S. Environmental Protection Agency (EPA) since 1972. Supplements to AP-42 have been routinely published to add new emission source categories and to update existing emission factor...

  11. APS controls overview

    SciTech Connect

    1996-02-01

    The APS accelerator control system described in this report is a distributed system consisting of operator interfaces, a network, and interfaces to hardware. The operator interface is a UNIX-based workstation with an X-windows graphical user interface. The workstation may be located at any point on the facility network and maintain full functionality. The user has the ability to generate and alter control displays and to access the alarm handler, the archiver, interactive control programs, custom code, and other tools. The TCP/EP networking protocol has been selected as the underlying protocol for the control system network. TCP/EP is a commercial standard and readily available from network hardware vendors. Its implementation is independent of the particular network medium selected to implement the controls network. In the development environment copper Ethernet is the network medium; however, in the actual implementation a fiber-based system using hub technology will be utilized. The function of the network is to provide a generalized communication path between the host computers, operator workstations, input/output crates, and other hardware that comprise the control system.

  12. Functional Analysis of AP-2 α and μ2 Subunits

    PubMed Central

    Motley, Alison M.; Berg, Nicola; Taylor, Marcus J.; Sahlender, Daniela A.; Hirst, Jennifer; Owen, David J.

    2006-01-01

    The AP-2 adaptor complex plays a key role in cargo recognition and clathrin-coated vesicle formation at the plasma membrane. To investigate the functions of individual binding sites and domains of the AP-2 complex in vivo, we have stably transfected HeLa cells with wild-type and mutant small interfering RNA–resistant α and μ2 subunits and then used siRNA knockdowns to deplete the endogenous proteins. Mutating the PtdIns(4,5)P2 binding site of α, the phosphorylation site of μ2, or the YXXΦ binding site of μ2 impairs AP-2 function, as assayed by transferrin uptake. In contrast, removing the C-terminal appendage domain of α, or mutating the PtdIns(4,5)P2 binding site of μ2, has no apparent effect. However, adding a C-terminal GFP tag to α renders it completely nonfunctional. These findings demonstrate that there is some functional redundancy in the binding sites of the various AP-2 subunits, because no single mutation totally abolishes function. They also help to explain why GFP-tagged AP-2 never appears to leave the plasma membrane in some live cell imaging studies. Finally, they establish a new model system that can be used both for additional structure-function analyses, and as a way of testing tagged constructs for function in vivo. PMID:17035630

  13. Final report for tank 241-AP-101, grab samples 1AP-95-1, 1AP-95-2, 1AP-95-3, 1AP-95-4, 1AP-95-5, and 1AP-95-6

    SciTech Connect

    Esch, R.A.

    1996-03-04

    Six supernate grab samples (1AP-95-1 through 6) and one field blank (1AP-95-7) were taken from tank 241-AP-101, on Nov. 10 and 13, 1995. Analyses were performed in support of the Safety Screening and the Waste Compatibility Safety programs. All analytical results were within the action limits stated in the TSAP.

  14. The AP-1 transcription factor homolog Pf-AP-1 activates transcription of multiple biomineral proteins and potentially participates in Pinctada fucata biomineralization

    PubMed Central

    Zheng, Xiangnan; Cheng, Minzhang; Xiang, Liang; Liang, Jian; Xie, Liping; Zhang, Rongqing

    2015-01-01

    Activator protein-1 (AP-1) is an important bZIP transcription factor that regulates a series of physiological processes by specifically activating transcription of several genes, and one of its well-chartered functions in mammals is participating in bone mineralization. We isolated and cloned the complete cDNA of a Jun/AP-1 homolog from Pinctada fucata and called it Pf-AP-1. Pf-AP-1 had a highly conserved bZIP region and phosphorylation sites compared with those from mammals. A tissue distribution analysis showed that Pf-AP-1 was ubiquitously expressed in P. fucata and the mRNA level of Pf-AP-1 is extremely high in mantle. Pf-AP-1 expression was positively associated with multiple biomineral proteins in the mantle. The luciferase reporter assay in a mammalian cell line showed that Pf-AP-1 significantly up-regulates the transcriptional activity of the promoters of KRMP, Pearlin, and Prisilkin39. Inhibiting the activity of Pf-AP-1 depressed the expression of multiple matrix proteins. Pf-AP-1 showed a unique expression pattern during shell regeneration and pearl sac development, which was similar to the pattern observed for biomineral proteins. These results suggest that the Pf-AP-1 AP-1 homolog is an important transcription factor that regulates transcription of several biomineral proteins simultaneously and plays a role in P. fucata biomineralization, particularly during pearl and shell formation. PMID:26404494

  15. Abasic phosphorothioate oligomers inhibit HIV-1 reverse transcription and block virus transmission across polarized ectocervical organ cultures.

    PubMed

    Fraietta, Joseph A; Mueller, Yvonne M; Lozenski, Karissa L; Ratner, Deena; Boesteanu, Alina C; Hancock, Aidan S; Lackman-Smith, Carol; Zentner, Isaac J; Chaiken, Irwin M; Chung, Suhman; LeGrice, Stuart F J; Snyder, Beth A; Mankowski, Marie K; Jones, Natalie M; Hope, Jennifer L; Gupta, Phalguni; Anderson, Sharon H; Wigdahl, Brian; Katsikis, Peter D

    2014-12-01

    In the absence of universally available antiretroviral (ARV) drugs or a vaccine against HIV-1, microbicides may offer the most immediate hope for controlling the AIDS pandemic. The most advanced and clinically effective microbicides are based on ARV agents that interfere with the earliest stages of HIV-1 replication. Our objective was to identify and characterize novel ARV-like inhibitors, as well as demonstrate their efficacy at blocking HIV-1 transmission. Abasic phosphorothioate 2' deoxyribose backbone (PDB) oligomers were evaluated in a variety of mechanistic assays and for their ability to inhibit HIV-1 infection and virus transmission through primary human cervical mucosa. Cellular and biochemical assays were used to elucidate the antiviral mechanisms of action of PDB oligomers against both lab-adapted and primary CCR5- and CXCR4-utilizing HIV-1 strains, including a multidrug-resistant isolate. A polarized cervical organ culture was used to test the ability of PDB compounds to block HIV-1 transmission to primary immune cell populations across ectocervical tissue. The antiviral activity and mechanisms of action of PDB-based compounds were dependent on oligomer size, with smaller molecules preventing reverse transcription and larger oligomers blocking viral entry. Importantly, irrespective of molecular size, PDBs potently inhibited virus infection and transmission within genital tissue samples. Furthermore, the PDB inhibitors exhibited excellent toxicity and stability profiles and were found to be safe for vaginal application in vivo. These results, coupled with the previously reported intrinsic anti-inflammatory properties of PDBs, support further investigations in the development of PDB-based topical microbicides for preventing the global spread of HIV-1. PMID:25224013

  16. Abasic Phosphorothioate Oligomers Inhibit HIV-1 Reverse Transcription and Block Virus Transmission across Polarized Ectocervical Organ Cultures

    PubMed Central

    Fraietta, Joseph A.; Mueller, Yvonne M.; Lozenski, Karissa L.; Ratner, Deena; Boesteanu, Alina C.; Hancock, Aidan S.; Lackman-Smith, Carol; Zentner, Isaac J.; Chaiken, Irwin M.; Chung, Suhman; LeGrice, Stuart F. J.; Snyder, Beth A.; Mankowski, Marie K.; Jones, Natalie M.; Hope, Jennifer L.; Gupta, Phalguni; Anderson, Sharon H.; Wigdahl, Brian

    2014-01-01

    In the absence of universally available antiretroviral (ARV) drugs or a vaccine against HIV-1, microbicides may offer the most immediate hope for controlling the AIDS pandemic. The most advanced and clinically effective microbicides are based on ARV agents that interfere with the earliest stages of HIV-1 replication. Our objective was to identify and characterize novel ARV-like inhibitors, as well as demonstrate their efficacy at blocking HIV-1 transmission. Abasic phosphorothioate 2′ deoxyribose backbone (PDB) oligomers were evaluated in a variety of mechanistic assays and for their ability to inhibit HIV-1 infection and virus transmission through primary human cervical mucosa. Cellular and biochemical assays were used to elucidate the antiviral mechanisms of action of PDB oligomers against both lab-adapted and primary CCR5- and CXCR4-utilizing HIV-1 strains, including a multidrug-resistant isolate. A polarized cervical organ culture was used to test the ability of PDB compounds to block HIV-1 transmission to primary immune cell populations across ectocervical tissue. The antiviral activity and mechanisms of action of PDB-based compounds were dependent on oligomer size, with smaller molecules preventing reverse transcription and larger oligomers blocking viral entry. Importantly, irrespective of molecular size, PDBs potently inhibited virus infection and transmission within genital tissue samples. Furthermore, the PDB inhibitors exhibited excellent toxicity and stability profiles and were found to be safe for vaginal application in vivo. These results, coupled with the previously reported intrinsic anti-inflammatory properties of PDBs, support further investigations in the development of PDB-based topical microbicides for preventing the global spread of HIV-1. PMID:25224013

  17. Delta-elimination by T4 endonuclease V at a thymine dimer site requires a secondary binding event and amino acid Glu-23.

    PubMed

    Latham, K A; Lloyd, R S

    1995-07-11

    Endonuclease V from bacteriophage T4 is a well characterized enzyme that initiates the repair of ultraviolet light induced pyrimidine dimers. Scission of the phosphodiester backbone between the pyrimidines within a dimer, or 3' to an abasic (AP) site, occurs by a beta-elimination mechanism. In addition, high concentrations of endonuclease V have been reported to catalyze the cleavage of the C5'-O-P bond in a reaction referred to as delta-elimination. To better understand the enzymology of endonuclease V, the delta-elimination reaction of the enzyme has been investigated using an oligonucleotide containing a site-specific cis-syn cyclobutane thymine dimer. The slower kinetics of the delta-elimination reaction compared to beta-elimination and the ability of unlabeled dimer-containing DNA to compete more efficiently for delta-elimination than beta-elimination indicate that delta-elimination most likely occurs during a separate enzyme encounter with the incised DNA. Previous studies have shown that both the alpha-amino group of the N-terminus and the acidic residue Glu-23 are necessary for the N-glycosylase and AP lyase activities of endonuclease V. Experiments with T2P, E23Q, and E23D mutants, which are defective in pyrimidine dimer-specific nicking, demonstrated that delta-elimination requires Glu-23, but not the primary amine at the N-terminus. In fact, the T2P mutant was much more efficient at promoting delta-elimination than the wild-type enzyme. Besides lending further proof that delta-elimination requires a second encounter between enzyme and DNA, this result may reflect an enhanced binding of the T2P mutant to dimer-containing DNA. PMID:7612620

  18. Meet the APS Journal Editors

    NASA Astrophysics Data System (ADS)

    2016-05-01

    The Editors of the APS journals invite you to join them for conversation. The Editors will be available to answer questions, hear your ideas, and discuss any comments about the journals. All are welcome. Light refreshments will be served.

  19. Variation in the wheat AP2 homoeologs, the genes underlying lodicule development.

    PubMed

    Ning, Shunzong; Wang, Ning; Sakuma, Shun; Pourkheirandish, Mohammad; Koba, Takato; Komatsuda, Takao

    2013-09-01

    The bread wheat genome harbors three homoeologs of the barley gene HvAP2, which determines the cleistogamous/non-cleistogamous flowering. The three homoeologs, TaAP2-A, TaAP2-B and TaAP2-D, are derived from the A, B and D genomes. The importance of lodicule swelling in assuring non-cleistogamous flowering in a range of wild and domesticated wheat accessions of varying ploidy level was established. Re-sequencing of wheat AP2 homoeologous genes was carried out to identify natural variation at both the nucleotide and polypeptide level. The sequences of wheat AP2 homoeologs are highly conserved even across different ploidy levels and no functional variants at the key miR172 targeting site were detected. These results indicate that engineering of cleistogamous wheat will require the presence of a functional TaAP2 modification at each of the three homoeologs. PMID:24273420

  20. AP Human Geography and Success on the AP Test

    ERIC Educational Resources Information Center

    Roncone, John; Newhalfen, Nate

    2013-01-01

    Classroom projects that explore culture and globalization enhance the curriculum and help students see how geography directly connects to their lives. These authors contend that a project-based approach can supplement the teaching of an AP Human Geography course, and visualize this course as an essential tool for students to truly understand how…

  1. Molecular Basis for the Interaction Between AP4 β4 and its Accessory Protein, Tepsin.

    PubMed

    Frazier, Meredith N; Davies, Alexandra K; Voehler, Markus; Kendall, Amy K; Borner, Georg H H; Chazin, Walter J; Robinson, Margaret S; Jackson, Lauren P

    2016-04-01

    The adaptor protein 4 (AP4) complex (ϵ/β4/μ4/σ4 subunits) forms a non-clathrin coat on vesicles departing the trans-Golgi network. AP4 biology remains poorly understood, in stark contrast to the wealth of molecular data available for the related clathrin adaptors AP1 and AP2. AP4 is important for human health because mutations in any AP4 subunit cause severe neurological problems, including intellectual disability and progressive spastic para- or tetraplegias. We have used a range of structural, biochemical and biophysical approaches to determine the molecular basis for how the AP4 β4 C-terminal appendage domain interacts with tepsin, the only known AP4 accessory protein. We show that tepsin harbors a hydrophobic sequence, LFxG[M/L]x[L/V], in its unstructured C-terminus, which binds directly and specifically to the C-terminal β4 appendage domain. Using nuclear magnetic resonance chemical shift mapping, we define the binding site on the β4 appendage by identifying residues on the surface whose signals are perturbed upon titration with tepsin. Point mutations in either the tepsin LFxG[M/L]x[L/V] sequence or in its cognate binding site on β4 abolish in vitro binding. In cells, the same point mutations greatly reduce the amount of tepsin that interacts with AP4. However, they do not abolish the binding between tepsin and AP4 completely, suggesting the existence of additional interaction sites between AP4 and tepsin. These data provide one of the first detailed mechanistic glimpses at AP4 coat assembly and should provide an entry point for probing the role of AP4-coated vesicles in cell biology, and especially in neuronal function. PMID:26756312

  2. A commercial PCV2a-based vaccine is effective in protection from experimental challenge of PCV2 mutant with two amino acids elongation in capsid protein.

    PubMed

    Guo, Long-Jun; Fu, Yu-Jie; Huang, Li-Ping; Wang, Yi-Ping; Wei, Yan-Wu; Wu, Hong-Li; Liu, Chang-Ming

    2015-07-17

    Current commercial PCV2 vaccines are almost based on PCV2a and have been shown to be effective in reducing PCV2a and PCV2b viremia and PCV2-associated lesions and diseases. The recent emergence of novel mutant PCV2 (mPCV2) strains and linkage of mPCV2 with cases of porcine circovirus associated disease (PCVAD) in pig herds have raised concerns over emergence of vaccine-escape mutants and reduced efficacy of PCV2a-based vaccines. The aim of this study was to determine the ability of a commercial PCV2a-based vaccine developed by our laboratory to protect conventional pigs against experimental challenge with mPCV2 at 9 weeks of age. Twenty 4-week-old pigs free of PCV2 infection were randomly divided into four treatment groups with 5 pigs each. Two groups were unvaccinated as positive and negative controls. Another two groups were vaccinated with the commercial PCV2a-based vaccine (PCV2-LG strain, China) at 4 weeks of age and identical booster immunization was conducted 3 weeks post primary immunization. At 9 weeks of age, all pigs except the negative control were challenged with a mutant PCV2b/YJ (mPCV2b/YJ) with two amino acids elongation in capsid protein. The experiment was terminated 28 days after challenge. Under the conditions of this study, vaccinated pigs were protected against PCV2 viremia and lesions whereas unvaccinated pigs were not. Moreover, mPCV2b/YJ infection was demonstrated in positive control and almost all had macroscopic or microscopic lesions consistent with PCVAD while negative control did not develop PCVAD. This study indicates that mPCV2b/YJ infection alone can trigger PCVAD development and that the commercial vaccine (PCV2-LG) is still effective in protecting conventional pigs against the emerging mPCV2b/YJ strain in China. PMID:26051516

  3. AP Geography, Environmental Science Thrive

    ERIC Educational Resources Information Center

    Robelen, Erik W.

    2012-01-01

    Geography may not be particularly known as a hot topic among today's students--even some advocates suggest it suffers from an image problem--but by at least one measure, the subject is starting to come into its own. Across more than 30 topics covered in the Advanced Placement (AP) program, participation in geography is rising faster than any…

  4. Coaching in the AP Classroom

    ERIC Educational Resources Information Center

    Fornaciari, Jim

    2013-01-01

    Many parallels exist between quality coaches and quality classroom teachers--especially AP teachers, who often feel the pressure to produce positive test results. Having developed a series of techniques and strategies for building a team-oriented winning culture on the field, Jim Fornaciari writes about how he adapted those methods to work in the…

  5. Transcription factor AP-2 regulates human immunodeficiency virus type 1 gene expression.

    PubMed Central

    Perkins, N D; Agranoff, A B; Duckett, C S; Nabel, G J

    1994-01-01

    Human immunodeficiency virus type 1 (HIV-1) gene expression is regulated by an enhancer region composed of multiple potential cis-acting regulatory sites. Here, we describe binding sites for the transcription factor AP-2 in the HIV-1 long terminal repeat which modulate HIV enhancer function. One site is embedded within the two previously described kappa B elements, and a second site is detected further downstream. DNase I footprinting and electrophoretic mobility shift assay experiments demonstrated that AP-2 binds to the site between the kappa B elements. Interestingly, AP-2 and NF-kappa B bind to this region in a mutually exclusive manner. Mutations which disrupt this AP-2-binding site lower basal levels of transcription but do not affect NF-kappa B-mediated induction by tumor necrosis factor alpha in Jurkat T leukemia cells. Images PMID:8084021

  6. Generation of a cre recombinase-conditional Nos1ap over-expression transgenic mouse

    PubMed Central

    Auer, Dallas R.; Sysa-Shah, Polina; Bedja, Djahida; Simmers, Jessica L.; Pak, Evgenia; Dutra, Amalia; Cohn, Ronald; Gabrielson, Kathleen L.

    2016-01-01

    Polymorphic non-coding variants at the NOS1AP locus have been associated with the common cardiac, metabolic and neurological traits and diseases. Although, in vitro gene targeting-based cellular and biochemical studies have shed some light on NOS1AP function in cardiac and neuronal tissue, to enhance our understanding of NOS1AP function in mammalian physiology and disease, we report the generation of cre recombinase-conditional Nos1ap over-expression transgenic mice (Nos1apTg). Conditional transgenic mice were generated by the pronuclear injection method and three independent, single-site, multiple copies integration event-based founder lines were selected. For heart-restricted over-expression, Nos1apTg mice were crossed with Mlc2v-cre and Nos1ap transcript over-expression was observed in left ventricles from Nos1apTg; Mlc2v-cre F1 mice. We believe that with the potential of conditional over-expression, Nos1apTg mice will be a useful resource in studying NOS1AP function in various tissues under physiological and disease states. PMID:24563304

  7. The APS control system network

    SciTech Connect

    Sidorowicz, K.V.; McDowell, W.P.

    1995-12-31

    The APS accelerator control system is a distributed system consisting of operator interfaces, a network, and computer-controlled interfaces to hardware. This implementation of a control system has come to be called the {open_quotes}Standard Model.{close_quotes} The operator interface is a UNDC-based workstation with an X-windows graphical user interface. The workstation may be located at any point on the facility network and maintain full functionality. The function of the network is to provide a generalized communication path between the host computers, operator workstations, input/output crates, and other hardware that comprise the control system. The crate or input/output controller (IOC) provides direct control and input/output interfaces for each accelerator subsystem. The network is an integral part of all modem control systems and network performance will determine many characteristics of a control system. This paper will describe the overall APS network and examine the APS control system network in detail. Metrics are provided on the performance of the system under various conditions.

  8. AP@home: The Artificial Pancreas Is Now at Home.

    PubMed

    Heinemann, Lutz; Benesch, Carsten; DeVries, J Hans

    2016-07-01

    In the past years the development of an artificial pancreas (AP) has made great progress and many activities are ongoing in this area of research. The major step forward made in the last years was moving the evaluation of AP systems from highly controlled experimental conditions to daily life conditions at the home of patients with diabetes; this was also the aim of the European Union-funded AP@home project. Over a time period of 5 years a series of clinical studies were performed that culminated in 2 "final studies" during which an AP system was used by patients in their home environment for 2 or 3 months without supervision by a physician, living their normal lives. Two different versions of the AP system developed within this project were evaluated. A significant improvement in glycated hemoglobin was observed during closed-loop conditions despite the fact that during the control period the patients used the best currently available therapeutic option. In addition, a "single-port AP system" was developed within the project that combines continuous glucose monitoring and insulin infusion at a single tissue site. By using such a combined device the patients not only have to carry one less device around, the number of access points through the skin is also reduced from 2 to 1. In summary, close cooperation of 12 European partners, both academic centers and industry, enabled the development and evaluation of AP systems under daily life conditions. The next step is to develop these into products in cooperation with commercial partners. PMID:26888971

  9. An AP Calculus Classroom Amusement Park

    ERIC Educational Resources Information Center

    Ferguson, Sarah

    2016-01-01

    Throughout the school year, AP Calculus teachers strive to teach course content comprehensively and swiftly in an effort to finish all required material before the AP Calculus exam. As early May approaches and the AP Calculus test looms, students and teachers nervously complete lessons, assignments, and assessments to ensure student preparation.…

  10. Preparing Students for the AP Psychology Exam

    ERIC Educational Resources Information Center

    Whitlock, Kristin

    2013-01-01

    The Advanced Placement Psychology exam is one of the fastest growing exams offered by the College Board. The average percent of change in the number of students taking this exam over the past five years is 12.4%. With 238,962 students taking the exam in 2013, the AP Psychology exam is the sixth largest exam, surpassing AP Biology and AP World…

  11. AP reclamation and reuse in RSRM propellant

    NASA Technical Reports Server (NTRS)

    Miks, Kathryn F.; Harris, Stacey A.

    1995-01-01

    A solid propellant ingredient reclamation pilot plant has been evaluated at the Strategic Operations of Thiokol Corporation, located in Brigham City, Utah. The plant produces AP wet cake (95 percent AP, 5 percent water) for recycling at AP vendors. AP has been obtained from two standard propellant binder systems (PBAN and HTPB). Analytical work conducted at Thiokol indicates that the vendor-recrystallized AP meets Space Shuttle propellant specification requirements. Thiokol has processed 1-, 5-, and 600-gallon propellant mixes with the recrystallized AP. Processing, cast, cure, ballistic, mechanical, and safety properties have been evaluated. Phillips Laboratory static-test-fired 70-pound and 800-pound BATES motors. The data indicate that propellant processed with reclaimed AP has nominal properties.

  12. APS high heat load monochromator

    SciTech Connect

    Lee, W.K.; Mills, D.

    1993-02-01

    This document contains the design specifications of the APS high heat load (HHL) monochromator and associated accessories as of February 1993. It should be noted that work is continuing on many parts of the monochromator including the mechanical design, crystal cooling designs, etc. Where appropriate, we have tried to add supporting documentation, references to published papers, and calculations from which we based our decisions. The underlying philosophy behind performance specifications of this monochromator was to fabricate a device that would be useful to as many APS users as possible, that is, the design should be as generic as possible. In other words, we believe that this design will be capable of operating on both bending magnet and ID beamlines (with the appropriate changes to the cooling and crystals) with both flat and inclined crystal geometries and with a variety of coolants. It was strongly felt that this monochromator should have good energy scanning capabilities over the classical energy range of about 4 to 20 keywith Si (111) crystals. For this reason, a design incorporating one rotation stage to drive both the first and second crystals was considered most promising. Separate rotary stages for the first and second crystals can sometimes provide more flexibility in their capacities to carry heavy loads (for heavily cooled first crystals or sagittal benders of second crystals), but their tuning capabilities were considered inferior to the single axis approach.

  13. Reduced repair capacity of a DNA clustered damage site comprised of 8-oxo-7,8-dihydro-2'-deoxyguanosine and 2-deoxyribonolactone results in an increased mutagenic potential of these lesions

    DOE PAGESBeta

    Cunniffe, Siobhan; O’Neill, Peter; Greenberg, Marc M.; Lomax, Martine E.

    2014-04-01

    A signature of ionizing radiation is the induction of DNA clustered damaged sites. Non-double strand break (DSB) clustered damage has been shown to compromise the base excision repair pathway, extending the lifetimes of the lesions within the cluster, compared to isolated lesions. This increases the likelihood the lesions persist to replication and thus increasing the mutagenic potential of the lesions within the cluster. Lesions formed by ionizing radiation include 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) and 2-deoxyribonolactone (dL). dL poses an additional challenge to the cell as it is not repaired by the short-patch base excision repair pathway. Here we show recalcitrant dL repairmore » is reflected in mutations observed when DNA containing it and a proximal 8-oxodGuo is replicated in Escherichia coli. 8-oxodGuo in close proximity to dL on the opposing DNA strand results in an enhanced frequency of mutation of the lesions within the cluster and a 20 base sequence flanking the clustered damage site in an E. coli based plasmid assay. In vitro repair of a dL lesion is reduced when compared to the repair of an abasic (AP) site and a tetrahydrofuran (THF), and this is due mainly to a reduction in the activity of polymerase β, leading to retarded FEN1 and ligase 1 activities. This study has given insights in to the biological effects of clusters containing dL.« less

  14. Physical identification of an internal promoter, ilvAp, in the distal portion of the ilvGMEDA operon.

    PubMed

    Lopes, J M; Lawther, R P

    1989-01-01

    It has been previously demonstrated that the ilvGMEDA operon is expressed in vivo from the promoters ilvGp2 and ilvEp. An additional internal promoter is identified and designated ilvAp. This internal promoter, which allows independent expression of ilvA, has been analyzed both in vivo and in vitro. Our results indicate that: (1) ilvAp exists in both Escherichia coli K-12 and Salmonella typhimurium, as demonstrated by fusion to the galK reporter gene; (2) ilvAp is located in the distal coding sequence of ilvD; (3) the ilvAp sequences are not identical for these two bacterial species; (4) transcription from ilvAp of E. coli K-12 was demonstrated; (5) expression from ilvAp responds to the availability of oxygen; (6) potential 3' 5'-cyclic AMP receptor protein binding sites exist adjacent to ilvAp. PMID:2473940

  15. Tank 241-AP-106, Grab samples, 6AP-98-1, 6AP-98-2 and 6AP-98-3 Analytical results for the final report

    SciTech Connect

    FULLER, R.K.

    1999-02-23

    This document is the final report for tank 241-AP-106 grab samples. Three grab samples 6AP-98-1, 6AP-98-2 and 6AP-98-3 were taken from riser 1 of tank 241-AP-106 on May 28, 1998 and received by the 222-S Laboratory on May 28, 1998. Analyses were performed in accordance with the ''Compatability Grab Sampling and Analysis Plan'' (TSAP) (Sasaki, 1998) and the ''Data Quality Objectives for Tank Farms Waste Compatability Program (DQO). The analytical results are presented in the data summary report. No notification limits were exceeded. The request for sample analysis received for AP-106 indicated that the samples were polychlorinated biphenyl (PCB) suspects. The results of this analysis indicated that no PCBs were present at the Toxic Substance Control Act (TSCA) regulated limit of 50 ppm. The results and raw data for the PCB analysis are included in this document.

  16. Thrombin induces endothelial arginase through AP-1 activation.

    PubMed

    Zhu, Weifei; Chandrasekharan, Unni M; Bandyopadhyay, Smarajit; Morris, Sidney M; DiCorleto, Paul E; Kashyap, Vikram S

    2010-04-01

    Arterial thrombosis is a common disease leading to severe ischemia beyond the obstructing thrombus. Additionally, endothelial dysfunction at the site of thrombosis can be rescued by l-arginine supplementation or arginase blockade in several animal models. Exposure of rat aortic endothelial cells (RAECs) to thrombin upregulates arginase I mRNA and protein levels. In this study, we further investigated the molecular mechanism of thrombin-induced arginase changes in endothelial cells. Thrombin strikingly increased arginase I promoter and enzyme activity in primary cultured RAECs. Using different deletion and point mutations of the promoter, we demonstrated that the activating protein-1 (AP-1) consensus site located at -3,157 bp in the arginase I promoter was a thrombin-responsive element. Electrophoretic mobility shift assay and chromatin immunoprecipitation assay further confirmed that upon thrombin stimulation, c-Jun and activating transcription factor-2 (ATF-2) bound to the AP-1 site, which initiated the transactivation. Moreover, loss-of-function studies using small interfering RNA confirmed that recruitment of these two transcription factors to the AP-1 site was required for thrombin-induced arginase upregulation. In the course of defining the signaling pathway leading to the activation of AP-1 by thrombin, we found thrombin-induced phosphorylation of stress-activated protein kinase/c-Jun-NH(2)-terminal kinase (SAPK/JNK or JNK1/2/3) and p38 mitogen-activated protein kinase, which were followed by the phosphorylation of both c-Jun and ATF-2. These findings reveal the basis for thrombin induction of endothelial arginase I and indicate that arginase inhibition may be an attractive therapeutic alternative in the setting of arterial thrombosis and its associated endothelial dysfunction. PMID:20032511

  17. Clathrin interactions with C-terminal regions of the yeast AP-1 beta and gamma subunits are important for AP-1 association with clathrin coats.

    PubMed

    Yeung, B G; Payne, G S

    2001-08-01

    Heterotetrameric adaptor (AP) complexes are thought to coordinate cargo recruitment and clathrin assembly during clathrin-coated vesicle biogenesis. We have identified, and characterized the physiological significance of clathrin-binding activities in the two large subunits of the AP-1 complex in Saccharomyces cerevisiae. Using GST-fusion chromatography, two clathrin-binding sites were defined in the beta1 subunit that match consensus clathrin-binding sequences in other mammalian and yeast clathrin-binding proteins. Clathrin interactions were also identified with the C-terminal region of the gamma subunit. When introduced into chromosomal genes, point mutations in the beta1 clathrin-binding motifs, or deletion of the gamma C-terminal region, reduced association of AP-1 with clathrin in coimmunoprecipitation assays. The beta1 mutations or the gamma truncation individually produced minor effects on AP-1 distribution by subcellular fractionation. However, when beta1 and gamma mutations were combined, severe defects were observed in AP-1 association with membranes and incorporation into clathrin-coated vesicles. The combination of subunit mutations accentuated growth and alpha-factor pheromone maturation defects in chc1-ts cells, though not to the extent caused by complete loss of AP-1 activity. Our results suggest that both the beta1 and gamma subunits contribute interactions with clathrin that are important for stable assembly of AP-1 complexes into clathrin coats in vivo. PMID:11489214

  18. ELECTROCHEMICAL CORROSION TESTING OF TANKS 241-AN-102 & 241-AP-107 & 241-AP-108 IN SUPPORT OF ULTRASONIC TESTING

    SciTech Connect

    WYRWAS RB; DUNCAN JB

    2008-11-20

    This report presents the results of the corrosion rates that were measured using electrochemical methods for tanks 241-AN-102 (AN-102), 241-AP-107 (AP 107), and 241-AP-108 (AP-108) performed under test plant RPP-PLAN-38215. The steel used as materials of construction for AN and AP tank farms was A537 Class 1. Test coupons of A537 Class 1 carbon steel were used for corrosion testing in the AN-107, AP-107, and AP-108 tank waste. Supernate will be tested from AN-102, AP-107, and Ap-108. Saltcake testing was performed on AP-108 only.

  19. Coaching Strategies for AP: Building a Successful AP European History Program

    ERIC Educational Resources Information Center

    Fornaciari, Jim

    2014-01-01

    The October 2013 special issue of "Social Education" dealt with almost all AP social studies subjects, but omitted AP European History. This is one of the most fascinating AP subjects for students and teachers alike. In this article, the author shares his experiences since hewas given the responsibility of building his school's…

  20. AP Music Theory in Your School.

    ERIC Educational Resources Information Center

    Lucia, Raymond

    1993-01-01

    Asserts that an Advanced Placement (AP) course in music theory offers student musicians opportunities to gain new insight into melody, harmony, and structure. Describes content and teaching methods used in an AP music theory program. Discusses necessary teacher characteristics and maintains that both students and teachers benefit from the course.…

  1. Toward lattice QCD simulation on AP1000

    NASA Astrophysics Data System (ADS)

    Ohta, Shigemi

    AP1000 is Fujitsu Laboratory's experimental parallel computer consisting of up to 1024 microcomputers called cells. It is found that each AP1000 cell can sustain two to three megaflops computational speed for full QCD lattice numerical simulations in IEEE 64-bit floating point format

  2. AP-42 ADDITIONS AND REVISIONS - WILDLAND FIRES

    EPA Science Inventory

    This project is aimed at consolidating, selecting, and disseminating the most appropriate data and methods for estimating air emissions from wildfires and prescribed burns. The product will replace a current section of AP-42, but may not take the precise form of an AP-42 secti...

  3. Advanced APS impacts on vehicle payloads

    NASA Astrophysics Data System (ADS)

    Schneider, Steven J.; Reed, Brian D.

    1989-04-01

    Advanced auxiliary propulsion system (APS) technology has the potential to both, increase the payload capability of earth-to-orbit (ETO) vehicles by reducing APS propellant mass, and simplify ground operations and logistics by reducing the number of fluids on the vehicle and eliminating toxic, corrosive propellants. The impact of integrated cryogenic APS on vehicle payloads is addressed. In this system, launch propulsion system residuals are scavenged from integral launch propulsion tanks for use in the APS. Sufficient propellant is preloaded into the APS to return to earth with margin and noncomplete scavenging assumed. No propellant conditioning is required by the APS, but ambient heat soak is accommodated. High temperature rocket materials enable the use of the unconditioned hydrogen/oxygen in the APS and are estimated to give APS rockets specific impulse of up to about 444 sec. The payload benefits are quantified and compared with an uprated monomethylhydrazine/nitrogen tetroxide system in a conservative fashion, by assuming a 25.5 percent weight growth for the hydrogen/oxygen system and a 0 percent weight growth for the uprated system. The combination of scavenging and high performance gives payload impacts which are highly mission specific. A payload benefit of 861 kg (1898 lbm) was estimated for a Space Station Freedom rendezvous mission and 2099 kg (4626 lbm) for a sortie mission, with payload impacts varying with the amount of launch propulsion residual propellants. Missions without liquid propellant scavenging were estimated to have payload penalties, however, operational benefits were still possible.

  4. AP Courses Get Audited for Quality

    ERIC Educational Resources Information Center

    Ashford, Ellie

    2007-01-01

    As the college admissions process has gotten much more competitive, the number of high school students taking Advanced Placement (AP) courses has soared. At the same time, policymakers and education leaders seek to get more minorities and students not on the college track to sign up for AP and other rigorous classes. But as high schools have…

  5. Advanced APS Impacts on Vehicle Payloads

    NASA Technical Reports Server (NTRS)

    Schneider, Steven J.; Reed, Brian D.

    1989-01-01

    Advanced auxiliary propulsion system (APS) technology has the potential to both, increase the payload capability of earth-to-orbit (ETO) vehicles by reducing APS propellant mass, and simplify ground operations and logistics by reducing the number of fluids on the vehicle and eliminating toxic, corrosive propellants. The impact of integrated cryogenic APS on vehicle payloads is addressed. In this system, launch propulsion system residuals are scavenged from integral launch propulsion tanks for use in the APS. Sufficient propellant is preloaded into the APS to return to earth with margin and noncomplete scavenging assumed. No propellant conditioning is required by the APS, but ambient heat soak is accommodated. High temperature rocket materials enable the use of the unconditioned hydrogen/oxygen in the APS and are estimated to give APS rockets specific impulse of up to about 444 sec. The payload benefits are quantified and compared with an uprated monomethyl hydrazine/nitrogen tetroxide system in a conservative fashion, by assuming a 25.5 percent weight growth for the hydrogen/oxygen system and a 0 percent weight growth for the uprated system. The combination and scavenging and high performance gives payload impacts which are highly mission specific. A payload benefit of 861 kg (1898 lbm) was estimated for a Space Station Freedom rendezvous mission and 2099 kg (4626 lbm) for a sortie mission, with payload impacts varying with the amount of launch propulsion residual propellants. Missions without liquid propellant scavenging were estimated to have payload penalties, however, operational benefits were still possible.

  6. Advanced APS impacts on vehicle payloads

    NASA Technical Reports Server (NTRS)

    Schneider, Steven J.; Reed, Brian D.

    1989-01-01

    Advanced auxiliary propulsion system (APS) technology has the potential to both, increase the payload capability of earth-to-orbit (ETO) vehicles by reducing APS propellant mass, and simplify ground operations and logistics by reducing the number of fluids on the vehicle and eliminating toxic, corrosive propellants. The impact of integrated cryogenic APS on vehicle payloads is addressed. In this system, launch propulsion system residuals are scavenged from integral launch propulsion tanks for use in the APS. Sufficient propellant is preloaded into the APS to return to earth with margin and noncomplete scavenging assumed. No propellant conditioning is required by the APS, but ambient heat soak is accommodated. High temperature rocket materials enable the use of the unconditioned hydrogen/oxygen in the APS and are estimated to give APS rockets specific impulse of up to about 444 sec. The payload benefits are quantified and compared with an uprated monomethylhydrazine/nitrogen tetroxide system in a conservative fashion, by assuming a 25.5 percent weight growth for the hydrogen/oxygen system and a 0 percent weight growth for the uprated system. The combination of scavenging and high performance gives payload impacts which are highly mission specific. A payload benefit of 861 kg (1898 lbm) was estimated for a Space Station Freedom rendezvous mission and 2099 kg (4626 lbm) for a sortie mission, with payload impacts varying with the amount of launch propulsion residual propellants. Missions without liquid propellant scavenging were estimated to have payload penalties, however, operational benefits were still possible.

  7. The status of APS, BESSRC, and NEET.

    SciTech Connect

    Ahmad, I.; Dunford, R. W.; Esbensen, H.; Gemmell, D. S.; Kanter, E. P.; Kraessig, B.; Rutt, U.; Southworth, S. H.

    1999-03-10

    We present a brief summary of the current status of the Advanced Photon Source (APS) at Argonne National Laboratory and of the facilities at two of the APS sectors operated by the Basic Energy Sciences Synchrotrons Radiation Center (BESSRC). This is followed by a report on recent measurements at BESSRC on the phenomenon of Nuclear Excitation by Electronic Transition (NEET).

  8. Combustion Response of AP Composite Propellants

    NASA Technical Reports Server (NTRS)

    Shusser, Michael; Cohen, Norman S.; Culick, E. C.

    2000-01-01

    The Cohen & Strand model for AP composite propellants is applied as boundary conditions, one for AP and one for binder, in solving the heat conduction equation in each to compute linear and non-linear combustion response properties for each and for the aggregate propellant. Iterations couple AP and binder through the quasi-steady flame processes. Illustrative results for linear response functions (pressure-coupled and velocity-coupled) are presented for a monomodal AP propellant varying AP size, pressure and crossflow speed, and examples of non-linear responses to arbitrary waveforms are shown. A quantitative comparison with response function data is limited to one well-characterized research formulation. Mechanistic implications are discussed, including recommendations for future modeling work.

  9. APS undulator radiation: First results

    SciTech Connect

    Cai, Z.; Dejus, R.J.; Hartog, P.D.

    1995-12-31

    The first undulator radiation has been extracted from the Advanced Photon Source (APS). The results from the characterization of this radiation are very satisfactory. With the undulator set at a gap of 15.8 mm (K=1.61), harmonics as high as the 17th were observed using a crystal spectrometer. The angular distribution of the third-harmonic radiation was measured, and the source was imaged using a zone plate to determine the particle beam emittance. The horizontal beam emittance was found to be 6.9 {plus_minus} 1.0 nm-rad, and the vertical emittance coupling was found to be less than 3%. The absolute spectral flux was measured over a wide range of photon energies, and it agrees remarkably well with the theoretical calculations based on the measured undulator magnetic field profile and the measured beam emittance. These results indicate that both the emittance of the electron beam and the undulator magnetic field quality exceed the original specifications.

  10. Status of APS 1-Mwe Parabolic Trough Project

    SciTech Connect

    Canada, S.; Brosseau, D.; Kolb, G.; Moore, L.; Cable, R.; Price, H.

    2005-11-01

    Arizona Public Service (APS) is currently installing new power facilities to generate a portion of its electricity from solar resources that will satisfy its obligation under the Arizona Environmental Portfolio Standard (EPS). During FY04, APS began construction on a 1-MWe parabolic trough concentrating solar power plant. This plant represents the first parabolic trough plant to begin construction since 1991. Site preparation and construction activities continued throughout much of FY05, and startup activities are planned for Fall 2005 (with completion early in FY06). The plant will be the first commercial deployment of the Solargenix parabolic trough collector technology developed under contract to the National Renewable Energy Laboratory. The plant will use an organic Rankine cycle (ORC) power plant, provided by Ormat. The ORC power plant is much simpler than the conventional steam Rankine cycle plant and allows unattended operation of the facility.

  11. AP1000{sup R} licensing and deployment in the United States

    SciTech Connect

    Jordan, R. P.; Russ, P. A.; Filiak, P. P.; Castiglione, L. L.

    2012-07-01

    In recent years, both domestic and foreign utilities have turned to the standardized Westinghouse AP1000 plant design in satisfying their near - and long-term - sustainable energy needs. As direct support to these actions, licensing the AP1000 design has played a significant role by providing one of the fundamental bases in clearing regulatory hurdles leading to the start of new plant construction. Within the U.S. alone, Westinghouse AP1000 licensing activities have reached unprecedented milestones with the approvals of both AP1000 Design Certification and Southern Company's combined construction permit and operating license (COL) application directly supporting the construction of two new nuclear plants in Georgia. Further COL application approvals are immediately pending for an additional two AP1000 plants in South Carolina. And, across the U.S. nuclear industry spectrum, there are 10 other COL applications under regulatory review representing some 16 new plants at 10 sites. In total, these actions represent the first wave of new plant licensing under the regulatory approval process since 1978. Fundamental to the Nuclear Regulatory Commission's AP1000 Design Certification is the formal recognition of the AP1000 passive safety design through regulatory acceptance rulemaking. Through recognition and deployment of the AP1000 Design Certification, the utility licensee / operator of this reactor design are now offered an opportunity to use a simplified 'one-step' combined license process, thereby managing substantial back-end construction schedule risk from regulatory and intervention delays. Application of this regulatory philosophy represents both acceptance and encouragement of standardized reactor designs like the AP1000. With the recent AP1000 Design Certification and utility COL acceptances, the fundamental licensing processes of this philosophy have successfully proven the attainment of significant milestones with the next stage licensing actions directed

  12. GSH1, which encodes gamma-glutamylcysteine synthetase, is a target gene for yAP-1 transcriptional regulation.

    PubMed Central

    Wu, A L; Moye-Rowley, W S

    1994-01-01

    Changes in gene dosage of the YAP1 gene, encoding the yAP-1 transcriptional regulatory protein, cause profound alterations in cellular drug and metal resistance. Previous studies on yAP-1 action in yeast cells have used the AP-1 response element (ARE) from simian virus 40 as an artificial site for yAP-1-mediated transcriptional activation. No authentic yeast target sites for control of gene expression by yAP-1 are known. Here we show that the GSH1 gene, encoding gamma-glutamylcysteine synthetase, is transcriptionally responsive to the yAP-1 protein. GSH1 encodes the rate-limiting step in yeast glutathione biosynthesis and contains within its promoter region a DNA element that matches the ARE in 11 of 12 positions. The GSH1 yAP-1 response element (YRE) was recognized by yAP-1 protein in vitro. Northern (RNA) blot analysis showed that GSH1 mRNA levels were responsive to YAP1 gene dosage. A site-directed mutation in the YRE that blocked yAP-1 binding in vitro prevented the mutant GSH1 promoter from responding to elevation in YAP1 gene dosage. A delta gsh1 mutant strain was constructed and unable to grow in the absence of exogenous glutathione. A mutant GSH1 gene lacking the YRE was unable to confer normal cadmium tolerance, although other yAP-1-mediated phenotypes remained normal. Thus, GSH1 is one of several genes that are transcriptionally controlled by yAP-1 and influence drug resistance. Images PMID:7915005

  13. APS Activities with Other Professional Societies

    NASA Astrophysics Data System (ADS)

    Slakey, Francis

    2006-03-01

    In 1981, the APS Council issued a statement that opposed ``equal time'' presentation in public school science classes of creationism and evolution. The statement clarified that ``Scientific inquiry and religious beliefs are two distinct elements of the human experience. Attempts to present them in the same context can only lead to misunderstandings of both.'' The APS Council revisited the issue in 1999 when a school board in Kansas attempted to eliminate the Big Bang, among other issues, from the science curriculum. Since that time, the APS has been more directly involved in confronting efforts that would dilute the teaching of science in public school science classes. This talk will review the APS activities and describe a developing multi-science society activity.

  14. (Very) Slow Rotation of Magnetic Ap Stars

    NASA Astrophysics Data System (ADS)

    Mathys, G.

    2015-04-01

    To this date, 33 magnetic Ap stars that have periods of variation longer than 30 days are known. They represent a considerable fraction of the total number of Ap stars whose period has been reliably determined. All the available evidence unambiguously indicates that the observed variations of those long-period Ap stars result from the changing aspect of their visible hemisphere as they rotate, thus that the oblique rotator model is applicable throughout the whole range of periods of variation of the Ap stars. We show that the periods of the most slowly rotating Ap stars must be of the order of 300 years, and that some may even be longer, possibly up to 1000 years. The 5 to 6 orders of magnitude spanned by the rotation periods of the Ap stars present a major challenge for the understanding of their origin and their evolution. To guide the theoretical developments, observational hints may be found in possible differences between the magnetic properties of stars that have rotation periods in different ranges. Such differences are starting to emerge from the existing data. To increase their significance level, study of the longest-period stars must be continued over their full rotation cycle. Failure to secure observations now may leave critical data missing for several decades, or even centuries.

  15. Human lung and bladder carcinoma tumors as compared to their adjacent normal tissue have elevated AP-1 activity associated with the retinoblastoma gene promoter.

    PubMed

    Linardopoulos, S; Papadakis, E; Delakas, D; Theodosiou, V; Cranidis, A; Spandidos, D A

    1993-01-01

    Examination of the nucleotide sequence of the retinoblastoma (Rb) promoter revealed the presence of a DNA region highly homologous to the recognition site for the cellular transcription factor AP-1. A pair of complementary oligonucleotides containing the AP-1 site was synthesized and used in gel retardation assays to determine the role of the AP-1 protein in the regulation of the Rb gene expression. Using nuclear extracts from Hela cells as well as from lung and bladder tumors, we found specific binding of the AP-1 protein to this oligonucleotide. This binding is elevated in Hela cells, in 10/13 lung and 3/8 bladder tumors as compared to adjacent normal tissue. These results suggest that AP-1 could be implicated in Rb gene transcriptional regulation through its interaction with the AP-1 binding site of the Rb gene promoter. PMID:8476221

  16. NOS1AP O-GlcNAc Modification Involved in Neuron Apoptosis Induced by Excitotoxicity.

    PubMed

    Zhu, Liang; Tao, Tao; Zhang, Dongmei; Liu, Xiaojuan; Ke, Kaifu; Shen, Aiguo

    2015-01-01

    O-Linked N-acetylglucosamine, or O-GlcNAc, is a dynamic post-translational modification that cycles on and off serine and threonine residues of nucleocytoplasmic and mitochondrial proteins. In addition to cancer and inflammation diseases, O-GlcNAc modification appears to play a critical role during cell apoptosis and stress response, although the precise mechanisms are still not very clear. Here we found that nitric oxide synthase adaptor (NOS1AP), which plays an important part in glutamate-induced neuronal apoptosis, carries the modification of O-GlcNAc. Mass spectrometry analysis identified Ser47, Ser183, Ser204, Ser269, Ser271 as O-GlcNAc sites. Higher O-GlcNAc of NOS1AP was detected during glutamate-induced neuronal apoptosis. Furthermore, with O-GlcNAc sites of NOS1AP mutated, the interaction of NOS1AP and neuronal nitric oxide syntheses (nNOS) decreases. Finally, during glutamate-induced neuronal apoptosis, decreasing the O-GlcNAc modification of NOS1AP results in more severe neuronal apoptosis. All these results suggest that O-GlcNAc modification of NOS1AP exerts protective effects during glutamate-induced neuronal apoptosis. PMID:26197318

  17. Structural determinants of DNA binding by a P. falciparum ApiAP2 transcriptional regulator

    PubMed Central

    Lindner, Scott E.; De Silva, Erandi K.; Keck, James L.; Llinás, Manuel

    2009-01-01

    Putative transcription factors have only recently been identified in the Plasmodium spp., with the major family of regulators comprising the Apicomplexan AP2 (ApiAP2) proteins. To better understand the DNA-binding mechanisms of these transcriptional regulators, we characterized the structure and in vitro function of an AP2 DNA-binding domain from a prototypical ApiAP2 protein, PF14_0633 from Plasmodium falciparum. The X-ray crystal structure of the PF14_0633 AP2 domain bound to DNA reveals a β-sheet fold that binds the DNA major groove through base-specific and backbone contacts; a prominent α-helix supports the β-sheet structure. Substitution of predicted DNA-binding residues with alanine weakened or eliminated DNA binding in solution. In contrast to plant AP2 domains, the PF14_0633 AP2 domain dimerizes upon binding to DNA through a domain-swapping mechanism in which the α-helices of the AP2 domains pack against the β-sheets of the dimer mates. DNA-induced dimerization of PF14_0633 may be important for tethering two distal DNA loci together in the nucleus and/or for inducing functional rearrangements of its domains to facilitate transcriptional regulation. Consistent with a multi-site binding mode, at least two copies of the consensus sequence recognized by PF14_0633 are present upstream of a previously identified group of sporozoite-stage genes. Taken together, these findings illustrate how Plasmodium has adapted the AP2 DNA-binding domain for genome-wide transcriptional regulation. PMID:19913037

  18. Thermodynamics of Damaged DNA Binding and Catalysis by Human AP Endonuclease 1.

    PubMed

    Miroshnikova, A D; Kuznetsova, A A; Kuznetsov, N A; Fedorova, O S

    2016-01-01

    Apurinic/apyrimidinic (AP) endonucleases play an important role in DNA repair and initiation of AP site elimination. One of the most topical problems in the field of DNA repair is to understand the mechanism of the enzymatic process involving the human enzyme APE1 that provides recognition of AP sites and efficient cleavage of the 5'-phosphodiester bond. In this study, a thermodynamic analysis of the interaction between APE1 and a DNA substrate containing a stable AP site analog lacking the C1' hydroxyl group (F site) was performed. Based on stopped-flow kinetic data at different temperatures, the steps of DNA binding, catalysis, and DNA product release were characterized. The changes in the standard Gibbs energy, enthalpy, and entropy of sequential specific steps of the repair process were determined. The thermodynamic analysis of the data suggests that the initial step of the DNA substrate binding includes formation of non-specific contacts between the enzyme binding surface and DNA, as well as insertion of the amino acid residues Arg177 and Met270 into the duplex, which results in the removal of "crystalline" water molecules from DNA grooves. The second binding step involves the F site flipping-out process and formation of specific contacts between the enzyme active site and the everted 5'-phosphate-2'-deoxyribose residue. It was shown that non-specific interactions between the binding surfaces of the enzyme and DNA provide the main contribution into the thermodynamic parameters of the DNA product release step. PMID:27099790

  19. Baseline water quality of Long Meadow Lake, Ponds AP-9 and AP-10, and Black Dog Creek, Hennepin and Dakota counties, Minnesota

    USGS Publications Warehouse

    Payne, G.A.

    1977-01-01

    Long Meadow Lake, Black Dog Creek, and Ponds AP-9 and AP-10 which lie in an area designated for a trunk highway bridge crossing the Minnesota River, were sampled for baseline water quality prior to construction of the bridge. Data collected show that dissolved solids fluctuate seasonally. Dissolved oxygen:/ranged from less than 1 milligram per liter under an ice cover to 13-9 milligrams per liter in the September sample at site 4 in Long Meadow Lake. The phytoplankton analyses showed pulses in algae populations, blue-green algae being the dominant type in at least one sample from each of the water courses.

  20. Simplified SBLOCA Analysis of AP1000

    SciTech Connect

    Brown, William L.

    2004-07-01

    The AP1000 is a 1000 MWe advanced nuclear power plant design that uses passive safety features such as a multi-stage, automatic depressurization system (ADS) and gravity-driven, safety injection from core make-up tanks (CMTs) and an in-containment refueling water storage tank (IRWST) to mitigate SBLOCA events. The period of most safety significance for AP1000 during a SBLOCA event is typically associated with the actuation of the fourth stage of the ADS and subsequent transition from CMT to IRWST safety injection. As this period of a SBLOCA is generally of a quasi-steady nature, the integral performance of the AP1000 can be understood and evaluated with a simplified model of the reactor vessel, ADS, and safety injection from the CMTs and IRWST. The simplified model of the AP1000 consists of a series of steady state simulations that uses drift flux in the core region and homogeneous treatment of the core exit region including the ADS flow paths to generate a family of core flow demand curves as a function of system pressure (i.e. mass flow required to satisfy core cooling). These core flow demand curves are plotted against passive safety system supply curves from the CMTs and IRWST to demonstrate the adequacy of the integral performance of the AP1000 during the most important phase of a SBLOCA. (author)

  1. Tank 241-AP-107, grab samples 7AP-97-1, 7AP-97-2 and 7AP-97-3 analytical results for the final report

    SciTech Connect

    Steen, F.H.

    1997-12-22

    This document is the final report for tank 241-AP-107 grab samples. Three grab samples were collected from riser 1 on September 11, 1997. Analyses were performed on samples 7AP-97-1, 7AP-97-2 and 7AP-97-3 in accordance with the Compatibility Grab Sampling and Analysis Plan (TSAP) (Sasaki, 1997) and the Data Quality Objectives for Tank Farms Waste Compatibility Program (DQO) (Rev. 1: Fowler, 1995; Rev. 2: Mulkey and Nuier, 1997). The analytical results are presented in the data summary report (Table 1). A notification was made to East Tank Farms Operations concerning low hydroxide in the tank and a hydroxide (caustic) demand analysis was requested. The request for sample analysis (RSA) (Attachment 2) received for AP-107 indicated that the samples were polychlorinated biphenyl (PCB) suspects. Therefore, prior to performing the requested analyses, aliquots were made to perform PCB analysis in accordance with the 222-S Laboratory administrative procedure, LAP-101-100. The results of this analysis indicated that no PCBs were present at 50 ppm and analysis proceeded as non-PCB samples. The results and raw data for the PCB analysis will be included in a revision to this document. The sample breakdown diagrams (Attachment 1) are provided as a cross-reference for relating the tank farm customer identification numbers with the 222-S Laboratory sample numbers and the portion of sample analyzed.

  2. Automated Plasma Spray (APS) process feasibility study

    NASA Technical Reports Server (NTRS)

    Fetheroff, C. W.; Derkacs, T.; Matay, I. M.

    1981-01-01

    An automated plasma spray (APS) process was developed to apply two layer (NiCrAlY and ZrO2-12Y2O3) thermal barrier coatings to aircraft and stationary gas turbine engine blade airfoils. The APS process hardware consists of four subsystems: a mechanical positioning subsystem incorporating two interlaced six degree of freedom assemblies (one for coating deposition and one for coating thickness monitoring); a noncoherent optical metrology subsystem (for in process gaging of the coating thickness buildup at specified points on the specimen); a microprocessor based adaptive system controller (to achieve the desired overall thickness profile on the specimen); and commerical plasma spray equipment. Over fifty JT9D first stage aircraft turbine blade specimens, ten W501B utility turbine blade specimens and dozens of cylindrical specimens were coated with the APS process in preliminary checkout and evaluation studies. The best of the preliminary turbine blade specimens achieved an overall coating thickness uniformity of 53 micrometers (2.1 mils), much better than is achievable manually. Comparative evaluations of coating thickness uniformity for manually sprayed and APS coated specimens were performed. One of the preliminary turbine blade evaluation specimens was subjected to a torch test and metallographic evaluation. Some cylindrical specimens coated with the APS process survived up to 2000 cycles in subsequent burner rig testing.

  3. AP-2{alpha} suppresses skeletal myoblast proliferation and represses fibroblast growth factor receptor 1 promoter activity

    SciTech Connect

    Mitchell, Darrion L.; DiMario, Joseph X.

    2010-01-15

    Skeletal muscle development is partly characterized by myoblast proliferation and subsequent differentiation into postmitotic muscle fibers. Developmental regulation of expression of the fibroblast growth factor receptor 1 (FGFR1) gene is required for normal myoblast proliferation and muscle formation. As a result, FGFR1 promoter activity is controlled by multiple transcriptional regulatory proteins during both proliferation and differentiation of myogenic cells. The transcription factor AP-2{alpha} is present in nuclei of skeletal muscle cells and suppresses myoblast proliferation in vitro. Since FGFR1 gene expression is tightly linked to myoblast proliferation versus differentiation, the FGFR1 promoter was examined for candidate AP-2{alpha} binding sites. Mutagenesis studies indicated that a candidate binding site located at - 1035 bp functioned as a repressor cis-regulatory element. Furthermore, mutation of this site alleviated AP-2{alpha}-mediated repression of FGFR1 promoter activity. Chromatin immunoprecipitation studies demonstrated that AP-2{alpha} interacted with the FGFR1 promoter in both proliferating myoblasts and differentiated myotubes. In total, these results indicate that AP-2{alpha} is a transcriptional repressor of FGFR1 gene expression during skeletal myogenesis.

  4. Building an AP Social Studies Program with Non-Traditional AP Students

    ERIC Educational Resources Information Center

    Ashmead, Amanda; Blanchette, Sue

    2013-01-01

    Equal access to education, that is to a high quality education, has increasingly come to mean access to an Advanced Placement program. In recent years, there has been steady attention paid to opening access to AP programs. The 9th annual College Board report (2013) stated "students who succeed on an AP Exam during high school typically…

  5. Laa1p, a Conserved AP-1 Accessory Protein Important for AP-1 Localization in Yeast

    PubMed Central

    Fernández, G. Esteban

    2006-01-01

    AP-1 and Gga adaptors participate in clathrin-mediated protein transport between the trans-Golgi network and endosomes. Both adaptors contain homologous domains that act to recruit accessory proteins involved in clathrin-coated vesicle formation, but the spectrum of known adaptor-binding partners is limited. This study describes an evolutionarily conserved protein of Saccharomyces cerevisiae, Laa1p (Yjl207cp), that interacts and functions specifically with AP-1. Deletion of LAA1, when combined with a conditional mutation in clathrin heavy chain or deletion of GGA genes, accentuated growth defects and increased disruption of clathrin-dependent α-factor maturation and transport of carboxypeptidase Y to the vacuole. In contrast, such genetic interactions were not observed between deletions of LAA1 and AP-1 subunit genes. Laa1p preferentially interacted with AP-1 compared with Gga proteins by glutathione S-transferase-fusion affinity binding and coimmunoprecipitations. Localization of AP-1 and Laa1p, but not Gga proteins, was highly sensitive to brefeldin A, an inhibitor of ADP-ribosylation factor (Arf) activation. Importantly, deletion of LAA1 caused mislocalization of AP-1, especially in cells at high density (postdiauxic shift), but it did not affect Gga protein distribution. Our results identify Laa1p as a new determinant of AP-1 localization, suggesting a model in which Laa1p and Arf cooperate to direct stable association of AP-1 with appropriate intracellular membranes. PMID:16687571

  6. Ectopic expression of FaesAP3, a Fagopyrum esculentum (Polygonaceae) AP3 orthologous gene rescues stamen development in an Arabidopsis ap3 mutant.

    PubMed

    Fang, Zheng-wu; Qi, Rui; Li, Xiao-fang; Liu, Zhi-xiong

    2014-10-25

    Arabidopsis thaliana APETALA3 (AP3) and Antirrhinum majus DEFICIENS (DEF) MADS box genes are required to specify petal and stamen identity. AP3 and DEF are members of the euAP3 lineage, which arose by gene duplication coincident with radiation of the core eudicots. In order to investigate the molecular mechanisms underlying organ development in early diverging clades of core eudicots, we isolated and identified an AP3 homolog, FaesAP3, from Fagopyrum esculentum (buckwheat, Polygonaceae), a multi-food-use pseudocereal with healing benefits. Protein sequence alignment and phylogenetic analyses revealed that FaesAP3 grouped into the euAP3 lineage. Expression analysis showed that FaesAP3 was transcribed only in developing stamens, and differed from AP3 and DEF, which expressed in developing petals and stamens. Moreover, ectopic expression of FaesAP3 rescued stamen development without complementation of petal development in an Arabidopsis ap3 mutant. Our results suggest that FaesAP3 is involved in the development of stamens in buckwheat. These results also suggest that FaesAP3 holds some potential for biotechnical engineering to create a male sterile line of F. esculentum. PMID:25149019

  7. Plyler Prize and APS Fellow Introductions

    NASA Astrophysics Data System (ADS)

    Nathanson, Gilbert

    2014-03-01

    The Division of Chemical Physics is delighted to announce the 2013 APS Fellows sponsored by DCP and to honor the 2014 Earl K. Plyler Prize Award winner. The new APS Fellows are: Ilan Benjamin, Hua Guo, Manos Mavrikakis, Josef Paldus, Joern Siepmann, Hans-Peter Steinrueck, Douglas Tobias, Angela Wilson, and Yijing Yan. The citations for each awardee will be read out loud. I will also introduce Prof. Lai-Sheng Wang of the Department of Chemistry at Brown University, who was awarded the 2014 Plyler Prize for Molecular Spectroscopy and Dynamics. Please come learn about these extraordinary scientists during this prize session. Prof. Wang's Plyler Prize talk will follow immediately after this introduction. For more information, see http://www.aps.org/units/dcp/.

  8. On the periodic variations of geomagnetic activity indices Ap and ap

    NASA Astrophysics Data System (ADS)

    Schreiber, H.

    1998-05-01

    Yearly averages of geomagnetic activity indices Ap for the years 1967-1984 are compared to the respective averages of 2·Bs, where v is the solar wind velocity and Bs is the southward interplanetary magnetic field (IMF) component. The correlation of both quantities is known to be rather good. Comparing the averages of Ap with 2 and Bs separately we find that, during the declining phase of the solar cycle, 2 and during the ascending phase Bs have more influence on Ap. According to this observation (using Fourier spectral analysis) the semiannual and 27 days, Ap variations for the years 1932-1993 were analysed separately for years before and after sunspot minima. Only those time-intervals before sunspot minima with a significant 27-day recurrent period of the IMF sector structure and those intervals after sunspot minima with a significant 28-28.5-day recurrent period of the sector structure were used. The averaged spectra of the two Ap data sets clearly show a period of 27 days before and a period of 28-29 days after sunspot minimum. Moreover, the phase of the average semiannual wave of Ap is significantly different for the two groups of data: the Ap variation maximizes near the equinoxes during the declining phase of the sunspot cycle and near the beginning of April and October during the ascending phase of the sunspot cycle, as predicted by the Russell-McPherron (R-M) mechanism. Analysing the daily variation of ap in an analogue manner, the same equinoctial and R-M mechanisms are seen, suggesting that during phases of the solar cycle, when ap depends more on the IMF-Bs component, the R-M mechanism is predominant, whereas during phases when ap increases as v increases the equinoctial mechanism is more likely to be effective.

  9. YY1 represses human papillomavirus type 16 transcription by quenching AP-1 activity.

    PubMed Central

    O'Connor, M J; Tan, S H; Tan, C H; Bernard, H U

    1996-01-01

    YY1 is a multifunctional transcription factor that has been shown to regulate the expression of a number of cellular and viral genes, including the human papillomavirus (HPV) oncogenes E6 and E7. In this study, we have analyzed the YY1-mediated repression of the HPV type 16 (HPV-16) E6-E7 promoter. A systematic analysis to identify YY1 sites present in the HPV-16 long control region showed that of 30 potential YY1 binding motifs, 24 bound purified recombinant YY1 protein, but only 10 of these were able to bind YY1 when nuclear extracts of HeLa cells were used. Of these, only a cluster of five sites, located in the vicinity of an AP-1 motif, were found to be responsible for repressing the HPV-16 P97 promoter. All five sites were required for repression, the mutation of any one site giving rise to a four- to sixfold increase in transcriptional activity. The target for YY1-mediated repression was identified as being a highly conserved AP-1 site, and we propose that AP-1 may represent a common target for YY1 repression. We also provide data demonstrating that YY1 can bind the transcriptional coactivator CREB-binding protein and propose a potentially novel mechanism by which YY1 represses AP-1 activity as a result of this YY1-CREB-binding protein interaction. PMID:8794287

  10. ap-9-(meta-tert-butylphenyl)fluorene.

    PubMed

    Robinson, Paul D; McLean, Aaron W; Meyers, Cal Y

    2003-10-01

    The title compound, C(23)H(22), (I), crystallizes in an ap conformationThe designations sp (synperiplanar) and ap (antiperiplanar) for these fluorene rotamers are in accordance with Rule E-6.6, IUPAC Tentative Rules, Section E, Fundamental Stereochemistry [J. Org. Chem. (1970), 35, 2861]. and its melt readily recrystallizes on cooling, in contrast to the corresponding 9-fluorenol compound, (II), which is sp and which melts without decomposition and fails to recrystallize over a long period. Both of these differences are ascribed to the intermolecular hydrogen bonding in (II), which is absent in (I) and which leads to distinctly different molecular packing in the two compounds. PMID:14532663

  11. Bcl-3 Expression Promotes Cell Survival following Interleukin-4 Deprivation and Is Controlled by AP1 and AP1-Like Transcription Factors

    PubMed Central

    Rebollo, Angelita; Dumoutier, Laure; Renauld, Jean-Christophe; Zaballos, Angel; Ayllón, Verónica; Martínez-A., Carlos

    2000-01-01

    We have analyzed the interleukin-4 (IL-4)-triggered mechanisms implicated in cell survival and show here that IL-4 deprivation induces apoptotic cell death but does not modulate Bcl-2 or Bcl-x expression. Since Bcl-x expression is insufficient to ensure cell survival in the absence of IL-4, we speculate that additional molecules replace the antiapoptotic role of Bcl-2 and Bcl-x in an alternative IL-4-triggered pathway. Cell death is associated with Bcl-3 downregulation and Bcl-3 expression blocks IL-4 deprivation-induced apoptosis, suggesting that Bcl-3 acts as a survival factor in the absence of growth factor. To characterize the IL-4-induced regulation of murine Bcl-3 expression, we cloned the promoter of this gene. Sequencing of the promoter showed no TATA box element but did reveal binding sites for AP1, AP1-like, and SP1 transcription factors. Retardation gels showed that IL-4 specifically induces AP1 and AP1-like binding activity and that mutation of these binding sites abolishes the IL-4-induced Bcl-3 promoter activity, suggesting that these transcription factors are important in Bcl-3 promoter transactivation. IL-4 deprivation induces downregulation of Jun expression and upregulation of Fos expression, both of which are proteins involved in the formation of AP1 and AP1-like transcription factors. Overexpression of Jun family proteins transactivates the promoter and restores Bcl-3 expression in the absence of IL-4 stimulation. Taken together, these data describe a new biological role for Bcl-3 and define the regulatory pathway implicated in Bcl-3 expression. PMID:10779330

  12. Suppression of albumin enhancer activity by H-ras and AP-1 in hepatocyte cell lines.

    PubMed Central

    Hu, J; Isom, H C

    1994-01-01

    We demonstrated, using a transient transfection assay, that the albumin enhancer increased the expression of the albumin promoter in a highly differentiated, simian virus 40 (SV40)-immortalized hepatocyte cell line, CWSV1, but was not functional in two ras-transformed cell lines (NR3 and NR4) derived from CWSV1 by stable transfection with the T24ras oncogene. A transient cotransfection assay showed that T24ras and normal c-Ha-ras were each able to inhibit the activity of the albumin enhancer in an immortal hepatocyte cell line. DNase I footprinting and gel mobility shift assays demonstrated that the DNA binding activities specific to the albumin enhancer were not decreased in the ras-transformed cells. ras also did not diminish the expression of HNF1 alpha, C/EBP alpha, HNF3 alpha, HNF3 beta, or HNF3 gamma but did significantly increase AP-1 binding activity. Three AP-1 binding sites were identified within the albumin enhancer, and DNA binding activities specific to these AP-1 sites were induced in the ras-transformed hepatocytes. Subsequent functional assays showed that overexpression of c-jun and c-fos inhibited the activity of the albumin enhancer. Site-directed mutagenesis of the AP-1 binding sites in the albumin enhancer partially abrogated the suppressing effect of ras and c-jun/c-fos on the enhancer. These functional studies therefore supported the results of the structural studies with AP-1. We conclude that the activity of the albumin enhancer is subject to regulation by ras signaling pathways and that the effect of ras on the albumin enhancer activity may be mediated by AP-1. Images PMID:8114691

  13. Structuring the AP Art History Course

    ERIC Educational Resources Information Center

    Herscher, Walter R.

    2013-01-01

    While AP (Advanced Placement) Art History may be taught within the art department in many schools, social studies teachers are equally capable of teaching the course well. They have the historical background to discuss the reasons for changes in art styles. A teacher's preparation is similar to teaching a course stressing political history,…

  14. APS deposition facility upgrades and future plans

    NASA Astrophysics Data System (ADS)

    Conley, Ray; Shi, Bing; Erdmann, Mark; Izzo, Scott; Assoufid, Lahsen; Goetze, Kurt; Mooney, Tim; Lauer, Kenneth

    2014-09-01

    The Advanced Photon Source (APS) has recently invested resources to upgrade or replace aging deposition systems with modern equipment. Of the three existing deposition systems, one will receive an upgrade, while two are being replaced. A design which adds a three-substrate planetary for the APS rotary deposition system is almost complete. The replacement for the APS large deposition system, dubbed the "Modular Deposition System", has been conceptually designed and is in the procurement process. Eight cathodes will sputter horizontally on mirrors up to 1.5 meters in length. This new instrument is designed to interface with ion-milling instruments and various metrology equipment for ion-beam figuring. A third linear machine, called the APS Profile Coating System, has two cathodes and is designed to accept substrates up to 200mm in length. While this machine is primarily intended for fabrication of figured KB mirrors using the profile-coating technique, it has also been used to produce multilayer monochromators for beamline use.

  15. The Promise of AP World History

    ERIC Educational Resources Information Center

    Saldaña, Cristóbal T.

    2013-01-01

    AP World History is the ideal history course. It introduces students to 10,000 years of world history, and demands critical reading, critical writing, and critical thinking skills on the part of both the teacher and the students. It requires students to build their expertise in reading their textbook, and places demands on the teacher to assign…

  16. Boosting Black Academic Achievement and AP Enrollments

    ERIC Educational Resources Information Center

    Saunders, Terrie; Maloney, Kathy

    2005-01-01

    Minority students were about 25% of the student population at Central High School in Omaha, Nebraska, in 1995. But the composition of its honors and challenge classes did not reflect this diversity: Few minority students were taking challenge classes as underclassmen and even fewer were taking Advanced Placement (AP) courses as seniors. This paper…

  17. College Board Readies Plans for AP Audits

    ERIC Educational Resources Information Center

    Klein, Alyson

    2006-01-01

    This article describes the educators mixed reviews regarding the audit system planned by the College Board to scrutinize high school Advanced Placement courses. Teachers of AP courses are required to submit materials to the College Board proving that their course syllabuses meet the program's curricular requirements. It is the most extensive…

  18. Two Successful Approaches to Teaching AP Government

    ERIC Educational Resources Information Center

    Ladd, Brian; Stepp, Heidi

    2013-01-01

    Amador Valley High School, in Pleasanton, California, uses two unique approaches to teaching Advanced Placement Government and Politics. AP Government consists of six units: Constitutional Underpinnings; Political Behavior and Political Beliefs; Mass Media, Interest Groups, and Political Parties; Institutions of Government; Civil Liberties and…

  19. The Demographic Wave: Rethinking Hispanic AP Trends

    ERIC Educational Resources Information Center

    Edwards, Kelcey; Sawtell, Ellen

    2013-01-01

    Presented at the Advanced Placement Annual Conference (APAC) in Las Vegas, NV in July 2013. This presentation reviews new research examining the AP® experience of Hispanic graduates over the past decade. Topics include an in-depth look at the AP Spanish Language and Culture gateway hypothesis and trends in family characteristics such as parent…

  20. The membrane-associated proteins FCHo and SGIP are allosteric activators of the AP2 clathrin adaptor complex

    PubMed Central

    Hollopeter, Gunther; Lange, Jeffrey J; Zhang, Ying; Vu, Thien N; Gu, Mingyu; Ailion, Michael; Lambie, Eric J; Slaughter, Brian D; Unruh, Jay R; Florens, Laurence; Jorgensen, Erik M

    2014-01-01

    The AP2 clathrin adaptor complex links protein cargo to the endocytic machinery but it is unclear how AP2 is activated on the plasma membrane. Here we demonstrate that the membrane-associated proteins FCHo and SGIP1 convert AP2 into an open, active conformation. We screened for Caenorhabditis elegans mutants that phenocopy the loss of AP2 subunits and found that AP2 remains inactive in fcho-1 mutants. A subsequent screen for bypass suppressors of fcho-1 nulls identified 71 compensatory mutations in all four AP2 subunits. Using a protease-sensitivity assay we show that these mutations restore the open conformation in vivo. The domain of FCHo that induces this rearrangement is not the F-BAR domain or the µ-homology domain, but rather is an uncharacterized 90 amino acid motif, found in both FCHo and SGIP proteins, that directly binds AP2. Thus, these proteins stabilize nascent endocytic pits by exposing membrane and cargo binding sites on AP2. DOI: http://dx.doi.org/10.7554/eLife.03648.001 PMID:25303366

  1. Functional description of APS beamline front ends

    SciTech Connect

    Kuzay, T.

    1993-02-01

    Traditional synchrotron sources were designed to produce bending magnet radiation and have proven to be an essential scientific tool. Currently, a new generation of synchrotron sources is being built that will be able to accommodate a large number of insertion device (ID) and high quality bending magnet (BM) sources. One example is the 7-GeV Advanced Photon Source (APS) now under construction at Argonne National Laboratory. The research and development effort at the APS is designed to fully develop the potential of this new generation of synchrotron sources. Of the 40 straight sections in the APS storage ring, 34 will be available for IDs. The remaining six sections are reserved for the storage ring hardware and diagnostics. Although the ring incorporates 80 BMs, only 40 of them can be used to extract radiation. The accelerator hardware shadows five of these 40 bending magnets, so the maximum number of BM sources on the lattice is 35. Generally, a photon beamline consists of four functional sections. The first section is the ID or the BM, which provides the radiation source. The second section, which is immediately outside the storage ring but inside a concrete shielding tunnel, is the front end, which is designed to control, define, and/or confine the x-ray beam. In the case of the APS, the front ends are designed to confine the photon beam. The third section, just outside the concrete shielding tunnel and on the experimental floor, is the first optics enclosure, which contains optics to filter and monochromatize the photon beam. The fourth section of a beamline consists of beam transports, additional optics, and experiment stations to do the scientific investigations. This document describes only the front ends of the APS beamlines.

  2. Mesh Oriented datABase

    SciTech Connect

    Tautges, Timothy J.

    2004-04-01

    MOAB is a component for representing and evaluating mesh data. MOAB can store stuctured and unstructured mesh, consisting of elements in the finite element "zoo". The functional interface to MOAB is simple yet powerful, allowing the representation of many types of metadata commonly found on the mesh. MOAB is optimized for efficiency in space and time, based on access to mesh in chunks rather than through individual entities, while also versatile enough to support individual entity access. The MOAB data model consists of a mesh interface instance, mesh entities (vertices and elements), sets, and tags. Entities are addressed through handles rather than pointers, to allow the underlying representation of an entity to change without changing the handle to that entity. Sets are arbitrary groupings of mesh entities and other sets. Sets also support parent/child relationships as a relation distinct from sets containing other sets. The directed-graph provided by set parent/child relationships is useful for modeling topological relations from a geometric model or other metadata. Tags are named data which can be assigned to the mesh as a whole, individual entities, or sets. Tags are a mechanism for attaching data to individual entities and sets are a mechanism for describing relations between entities; the combination of these two mechanisms isa powerful yet simple interface for representing metadata or application-specific data. For example, sets and tags can be used together to describe geometric topology, boundary condition, and inter-processor interface groupings in a mesh. MOAB is used in several ways in various applications. MOAB serves as the underlying mesh data representation in the VERDE mesh verification code. MOAB can also be used as a mesh input mechanism, using mesh readers induded with MOAB, or as a t’anslator between mesh formats, using readers and writers included with MOAB.

  3. Mesh Oriented datABase

    2004-04-01

    MOAB is a component for representing and evaluating mesh data. MOAB can store stuctured and unstructured mesh, consisting of elements in the finite element "zoo". The functional interface to MOAB is simple yet powerful, allowing the representation of many types of metadata commonly found on the mesh. MOAB is optimized for efficiency in space and time, based on access to mesh in chunks rather than through individual entities, while also versatile enough to support individualmore » entity access. The MOAB data model consists of a mesh interface instance, mesh entities (vertices and elements), sets, and tags. Entities are addressed through handles rather than pointers, to allow the underlying representation of an entity to change without changing the handle to that entity. Sets are arbitrary groupings of mesh entities and other sets. Sets also support parent/child relationships as a relation distinct from sets containing other sets. The directed-graph provided by set parent/child relationships is useful for modeling topological relations from a geometric model or other metadata. Tags are named data which can be assigned to the mesh as a whole, individual entities, or sets. Tags are a mechanism for attaching data to individual entities and sets are a mechanism for describing relations between entities; the combination of these two mechanisms isa powerful yet simple interface for representing metadata or application-specific data. For example, sets and tags can be used together to describe geometric topology, boundary condition, and inter-processor interface groupings in a mesh. MOAB is used in several ways in various applications. MOAB serves as the underlying mesh data representation in the VERDE mesh verification code. MOAB can also be used as a mesh input mechanism, using mesh readers induded with MOAB, or as a t’anslator between mesh formats, using readers and writers included with MOAB.« less

  4. Tank 241-AP-106, grab samples, 6AP-96-1 through 6AP-96-3 analytical results for the final report

    SciTech Connect

    Esch, R.A., Westinghouse Hanford

    1996-12-11

    This document is the final report for tank 241-AP-106 grab samples. This document presents the analytical results for three samples (6AP-96-1, 6AP-96-2 and 6AP-96-3) taken from riser 1 @ 150{degrees} of tank 241-AP-1 06 on September 12, 1996. Analyses were performed in accordance with the Compatibility Grab Sampling and Analysis Plan (TSAP) (Sasaki, 1996) and the Data Quality Objectives for Tank Farms Waste Compatibility Program (DQO) (Fowler, 1995).

  5. Generic effluent monitoring system certification for AP-40 exhauster stack

    SciTech Connect

    Glissmeyer, J.A.; Davis, W.E.; Bussell, J.H.; Maughan, A.D.

    1997-09-01

    Tests were conducted to verify that the Generic Effluent Monitoring System (GEMS), as applied to the AP-40 exhauster stack, meets all applicable regulatory performance criteria for air sampling systems at nuclear facilities. These performance criteria address both the suitability of the air sampling probe location and the transport of the sample to the collection devices. The criteria covering air sampling probe location ensure that the contaminants in the stack are well mixed with the airflow at the probe location such that the extracted sample represents the whole. The sample transport criteria ensure that the sampled contaminants are quantitatively delivered to the collection device. The specific performance criteria are described in detail in the report. The tests demonstrated that the GEMS/AP-40 system meets all applicable performance criteria. The contaminant mixing tests were conducted by Pacific Northwest National Laboratory (PNNL) at the wind tunnel facility, 331-H Building, using a mockup of the actual stack. The particle sample transport tests were conducted by PNNL at the Numatec Hanford Company`s 305 Building. The AP-40 stack is typical of several 10-in. diameter stacks that discharge the filtered ventilation air from tank farms at the U.S. Department of Energy`s Hanford Site in Richland, Washington. The GEMS design features a probe with a single shrouded sampling nozzle, a sample delivery line, and sample collection system. The collection system includes a filter holder to collect the sample of record and an in-line detector head and filter for monitoring beta radiation-emitting particles. Unrelated to the performance criteria, it was found that the record sample filter holder exhibited symptoms of sample bypass around the particle collection filter. This filter holder should either be modified or replaced with a different type. 10 refs., 8 figs., 6 tabs.

  6. Modulation of KvAP Unitary Conductance and Gating by 1-Alkanols and Other Surface Active Agents

    PubMed Central

    Finol-Urdaneta, Rocio K.; McArthur, Jeffrey R.; Juranka, Peter F.; French, Robert J.; Morris, Catherine E.

    2010-01-01

    Abstract The actions of alcohols and anesthetics on ion channels are poorly understood. Controversy continues about whether bilayer restructuring is relevant to the modulatory effects of these surface active agents (SAAs). Some voltage-gated K channels (Kv), but not KvAP, have putative low affinity alcohol-binding sites, and because KvAP structures have been determined in bilayers, KvAP could offer insights into the contribution of bilayer mechanics to SAA actions. We monitored KvAP unitary conductance and macroscopic activation and inactivation kinetics in PE:PG/decane bilayers with and without exposure to classic SAAs (short-chain 1-alkanols, cholesterol, and selected anesthetics: halothane, isoflurane, chloroform). At levels that did not measurably alter membrane specific capacitance, alkanols caused functional changes in KvAP behavior including lowered unitary conductance, modified kinetics, and shifted voltage dependence for activation. A simple explanation is that the site of SAA action on KvAP is its entire lateral interface with the PE:PG/decane bilayer, with SAA-induced changes in surface tension and bilayer packing order combining to modulate the shape and stability of various conformations. The KvAP structural adjustment to diverse bilayer pressure profiles has implications for understanding desirable and undesirable actions of SAA-like drugs and, broadly, predicts that channel gating, conductance and pharmacology may differ when membrane packing order differs, as in raft versus nonraft domains. PMID:20197029

  7. Action of a mammalian AP-endonuclease on DNAs of defined sequences.

    PubMed Central

    Haukanes, B I; Helland, D E; Kleppe, K

    1989-01-01

    An apurinic/apyrimidinic (AP) specific endonuclease from mouse plasmacytoma cells (line MPC-11), was observed to cleave apurinic sites in oligonucleotides 9, 11, 12, 39 and 40 nucleotides in length. However, the enzyme failed to cleave AP-sites in two oligonucleotides 7 nucleotides in length. The maximum rates of digestion observed on these short single-stranded DNA (ssDNA) fragments were approximately 1/30 of the rates observed on double-stranded DNA (dsDNA). In studies using the Maxam-Gilbert DNA sequencing analysis, apurinic sites in purine-rich regions were preferentially cleaved in dsDNA but not in ssDNA, indicating that the enzyme has a sequence preference on dsDNA. These results suggest that some sites on DNA might be more efficiently repaired than others. Images PMID:2466239

  8. QM/MM analysis suggests that Alkaline Phosphatase (AP) and Nucleotide pyrophosphatase/phosphodiesterase slightly tighten the transition state for phosphate diester hydrolysis relative to solution: implication for catalytic promiscuity in the AP superfamily

    PubMed Central

    Hou, Guanhua

    2011-01-01

    Several members of the Alkaline Phosphatase (AP) superfamily exhibit a high level of catalytic proficiency and promiscuity in structurally similar active sites. A thorough characterization of the nature of transition state for different substrates in these enzymes is crucial for understanding the molecular mechanisms that govern those remarkable catalytic properties. In this work, we study the hydrolysis of a phosphate diester, MpNPP−, in solution, two experimentally well-characterized variants of AP (R166S AP, R166S/E322Y AP) and wild type Nucleotide pyrophosphatase/phosphodiesterase (NPP) by QM/MM calculations in which the QM method is an approximate density functional theory previously parameterized for phosphate hydrolysis (SCC-DFTBPR). The general agreements found between these calculations and available experimental data for both solution and enzymes support the use of SCC-DFTBPR/MM for a semi-quantitative analysis of the catalytic mechanism and nature of transition state in AP and NPP. Although phosphate diesters are cognate substrates for NPP but promiscuous substrates for AP, the calculations suggest that their hydrolysis reactions catalyzed by AP and NPP feature similar synchronous transition states that are slightly tighter in nature compared to that in solution, due in part to the geometry of the bimetallic zinc motif. Therefore, this study provides the first direct computational support to the hypothesis that enzymes in the AP superfamily catalyze cognate and promiscuous substrates via similar transition states to those in solution. Our calculations do not support the finding of recent QM/MM studies by López-Canut and coworkers, who suggested that the same diester substrate goes through a much looser transition state in NPP/AP than in solution, a result likely biased by the large structural distortion of the bimetallic zinc site in their simulations. Finally, our calculations for different phosphate diester orientations and phosphorothioate diesters

  9. Transcriptional regulation of endothelial nitric oxide synthase expression in uterine artery endothelial cells by c-Jun/AP-1

    PubMed Central

    Qian, Xiao-Xian; Mata-Greenwood, Eugenia; Liao, Wu Xiang; Zhang, Honghai; Zheng, Jing; Chen, Dong-bao

    2007-01-01

    Despite extensive studies have shown that increased endothelial nitric oxide synthase (NOS3) expression in the uterine artery endothelial cells (UAEC) plays a key role in uterine vasodilatation, the molecular mechanism controlling NOS3 expression in UAEC is unknown. According to the sheep NOS3 promoter sequence isolated in our laboratory, we hypothesize that the activator protein-1 (AP-1) site in the proximal sheep NOS3 promoter (TGAGTCA, -682 to -676) is important for NOS3 expression. We developed a c-Jun adenoviral expression system to overexpress c-Jun protein into UAEC to investigate the effects of c-Jun/AP-1 on NOS3 expression. Basal levels of c-Jun protein and mRNA were detected in UAEC. C-Jun protein was overexpressed in a concentration and time-dependent fashion in UAEC infected with sense c-Jun (S-c-Jun), but not sham and antisense c-Jun (A-c-Jun) adenoviruses. Infection with S-c-Jun adenovirus (25 MOI, multiplicity of infection) resulted in efficient c-Jun protein overexpression in UAEC up to 3 days. In S-c-Jun, but not sham and A-c-Jun adenovirus infected UAEC, NOS3 mRNA and protein levels were increased (P<0.05) compared to noninfected controls. Increased NOS3 expression was associated with increased total NOS activity. Transient transfections showed that c-Jun overexpression augmented the transactivation of the sheep NOS3 promoter-driven luciferase/reporter constructs with the AP-1 site but not of deletion constructs without the AP-1 site. When the AP-1 site was mutated, c-Jun failed to trans-activate the sheep NOS3 promoter. AP-1 DNA binding activity also increased in c-Jun overexpressed UAEC. Lastly, the pharmacological AP-1 activator phorbol myristate acetate increased AP-1 binding, trans-activated the wild-type but not the AP-1 mutant NOS3 promoter and dose-dependently stimulated UAEC NOS3 and c-Jun protein expression. Hence, our data show that c-Jun/AP-1 regulates NOS3 transcription involving the proximal AP-1 site in the 5′-regulatory region of

  10. Cultural Diversity in AP Art History

    ERIC Educational Resources Information Center

    Bolte, Frances R.

    2006-01-01

    Teaching AP Art History is like running on a treadmill that is moving faster than a teacher can run. Many teachers are out of breath before the end of the term and wonder how in the world they can cover every chapter. Because time is short and art from pre-history through to the present, including the non-European traditions, must be covered, this…

  11. An Updated AP2 Beamline TURTLE Model

    SciTech Connect

    Gormley, M.; O'Day, S.

    1991-08-23

    This note describes a TURTLE model of the AP2 beamline. This model was created by D. Johnson and improved by J. Hangst. The authors of this note have made additional improvements which reflect recent element and magnet setting changes. The magnet characteristics measurements and survey data compiled to update the model will be presented. A printout of the actual TURTLE deck may be found in appendix A.

  12. Beryllium window for an APS diagnostics beamline

    SciTech Connect

    Sheng, I.C.; Yang, B.X.; Sharma, Y.S.

    1997-09-01

    A beryllium (Be) window for an Advanced Photon Source (APS) diagnostics beamline has been designed and built. The window, which has a double concave axisymmetrical profile with a thickness of 0.5 mm at the center, receives 160 W/mm{sup 2} (7 GeV/100 mA stored beam) from an undulator beam. The window design as well as thermal and thermomechanical analyses, including thermal buckling of the Be window, are presented.

  13. APS storage ring vacuum system performance

    SciTech Connect

    Noonan, J.R.; Gagliano, J.; Goeppner, G.A.

    1997-06-01

    The Advanced Photon Source (APS) storage ring was designed to operated with 7-GeV, 100-mA positron beam with lifetimes > 20 hours. The lifetime is limited by residual gas scattering and Touschek scattering at this time. Photon-stimulated desorption and microwave power in the rf cavities are the main gas loads. Comparison of actual system gas loads and design calculations will be given. In addition, several special features of the storage ring vacuum system will be presented.

  14. Final report for tank 241-AP-108, grab samples 8AP-96-1, 8AP-96-2 and 8AP-96-FB

    SciTech Connect

    Esch, R.A.

    1996-04-19

    This document is the final report deliverable for the tank 241-AP-108 grab samples. The samples were subsampled and analyzed in accordance with the TSAP. Included in this report are the results for the Waste Compatibility analyses, with the exception of DSC and thermogravimetric analysis (TGA) results which were presented in the 45 Day report (Part 2 of this document). The raw data for all analyses, with the exception of DSC and TGA, are also included in this report.

  15. AP-42 ADDITIONS AND REVISIONS - RESIDENTIAL WOOD COMBUSTION

    EPA Science Inventory

    This project develops emission factors, etc., for residential fireplaces and woodstoves which are incorporated into AP-42. AP42 is a massive collection of material which describes processes which generate air emissions and presents emission factors and control effectiveness infor...

  16. Small Molecule Inhibitors Targeting Activator Protein 1 (AP-1)

    PubMed Central

    2015-01-01

    Activator protein 1 (AP-1) is a pivotal transcription factor that regulates a wide range of cellular processes including proliferation, apoptosis, differentiation, survival, cell migration, and transformation. Accumulating evidence supports that AP-1 plays an important role in several severe disorders including cancer, fibrosis, and organ injury, as well as inflammatory disorders such as asthma, psoriasis, and rheumatoid arthritis. AP-1 has emerged as an actively pursued drug discovery target over the past decade. Excitingly, a selective AP-1 inhibitor T-5224 (51) has been investigated in phase II human clinical trials. Nevertheless, no effective AP-1 inhibitors have yet been approved for clinical use. Despite significant advances achieved in understanding AP-1 biology and function, as well as the identification of small molecules modulating AP-1 associated signaling pathways, medicinal chemistry efforts remain an urgent need to yield selective and efficacious AP-1 inhibitors as a viable therapeutic strategy for human diseases. PMID:24831826

  17. DNA Apurinic-Apyrimidinic Site Binding And Excision By Endonuclease IV

    SciTech Connect

    Garcin, E.D.; Hosfield, D.J.; Desai, S.A.; Haas, B.J.; Bjoras, M.; Cunningham, R.P.; Tainer, J.A.

    2009-05-18

    Escherichia coli endonuclease IV is an archetype for an abasic or apurinic-apyrimidinic endonuclease superfamily crucial for DNA base excision repair. Here biochemical, mutational and crystallographic characterizations reveal a three-metal ion mechanism for damage binding and incision. The 1.10-{angstrom} resolution DNA-free and the 2.45-{angstrom} resolution DNA-substrate complex structures capture substrate stabilization by Arg37 and reveal a distorted Zn{sub 3}-ligand arrangement that reverts, after catalysis, to an ideal geometry suitable to hold rather than release cleaved DNA product. The 1.45-{angstrom} resolution DNA-product complex structure shows how Tyr72 caps the active site, tunes its dielectric environment and promotes catalysis by Glu261-activated hydroxide, bound to two Zn{sup 2+} ions throughout catalysis. These structural, mutagenesis and biochemical results suggest general requirements for abasic site removal in contrast to features specific to the distinct endonuclease IV alpha-beta triose phosphate isomerase (TIM) barrel and APE1 four-layer alpha-beta folds of the apurinic-apyrimidinic endonuclease families.

  18. Bivalent Motif-Ear Interactions Mediate the Association of the Accessory Protein Tepsin with the AP-4 Adaptor Complex.

    PubMed

    Mattera, Rafael; Guardia, Carlos M; Sidhu, Sachdev S; Bonifacino, Juan S

    2015-12-25

    The heterotetrameric (ϵ-β4-μ4-σ4) complex adaptor protein 4 (AP-4) is a component of a non-clathrin coat involved in protein sorting at the trans-Golgi network (TGN). Considerable interest in this complex has arisen from the recent discovery that mutations in each of its four subunits are the cause of a congenital intellectual disability and movement disorder in humans. Despite its physiological importance, the structure and function of this coat remain poorly understood. To investigate the assembly of the AP-4 coat, we dissected the determinants of interaction of AP-4 with its only known accessory protein, the ENTH/VHS-domain-containing protein tepsin. Using a variety of protein interaction assays, we found that tepsin comprises two phylogenetically conserved peptide motifs, [GS]LFXG[ML]X[LV] and S[AV]F[SA]FLN, within its C-terminal unstructured region, which interact with the C-terminal ear (or appendage) domains of the β4 and ϵ subunits of AP-4, respectively. Structure-based mutational analyses mapped the binding site for the [GS]LFXG[ML]X[LV] motif to a conserved, hydrophobic surface on the β4-ear platform fold. Both peptide-ear interactions are required for efficient association of tepsin with AP-4, and for recruitment of tepsin to the TGN. The bivalency of the interactions increases the avidity of tepsin for AP-4 and may enable cross-linking of multiple AP-4 heterotetramers, thus contributing to the assembly of the AP-4 coat. In addition to revealing critical aspects of this coat, our findings extend the paradigm of peptide-ear interactions, previously established for clathrin-AP-1/AP-2 coats, to a non-clathrin coat. PMID:26542808

  19. Accessing, Mining, and Archiving an On-line Database -- The APS Catalog of the POSS I

    NASA Astrophysics Data System (ADS)

    Humphreys, R. M.; Cabanela, J. E.; Kriessler, J.

    2000-12-01

    The APS Catalog of the POSS I is an on-line database of over 100 million stars and galaxies (http://aps.umn.edu). A unique subset of this database with over 218,000 galaxies within 30 degrees of the North Galactic Pole, the MAPS-NGP, is now available at our web site. This diameter--selected catalog (>= 10 arcsec) is the deepest galaxy catalog constructed over such a large area of the sky (3000 sq. degrees). The MAPS-NGP includes many additional parameters for the galaxy images not available in the APS Catalog. Working with members of our computer science department, we have developed a morphological classifier for galaxies that divides our galaxy type into three classes -- early, intermediate, and late. We have applied data mining techniques to identify the most useful image parameters for input into a neural network and decision--tree based classifier pipeline. We are also archiving the APS Catalog for distribution to astronomical data centers including NASA's ADC and SIMBAD at CDS. The extragalactic subset will be integrated into the NASA/IPAC extragalactic database(NED). The MAPS-NGP has already been provided to NED. The APS is supported by NASA's Applied Information Systems Research Program.

  20. Sorting of the Alzheimer's Disease Amyloid Precursor Protein Mediated by the AP-4 Complex

    SciTech Connect

    Burgos, Patricia V.; Mardones, Gonzalo A.; Rojas, Adriana L.; daSilva, Luis L.P.; Prabhu, Yogikala; Hurley, James H.; Bonifacino, Juan S.

    2010-08-12

    Adaptor protein 4 (AP-4) is the most recently discovered and least well-characterized member of the family of heterotetrameric adaptor protein (AP) complexes that mediate sorting of transmembrane cargo in post-Golgi compartments. Herein, we report the interaction of an YKFFE sequence from the cytosolic tail of the Alzheimer's disease amyloid precursor protein (APP) with the {micro}4 subunit of AP-4. Biochemical and X-ray crystallographic analyses reveal that the properties of the APP sequence and the location of the binding site on 4 are distinct from those of other signal-adaptor interactions. Disruption of the APP-AP-4 interaction decreases localization of APP to endosomes and enhances {gamma}-secretase-catalyzed cleavage of APP to the pathogenic amyloid-{beta} peptide. These findings demonstrate that APP and AP-4 engage in a distinct type of signal-adaptor interaction that mediates transport of APP from the trans-Golgi network (TGN) to endosomes, thereby reducing amyloidogenic processing of the protein.

  1. A Novel Sequence in AP180 and CALM Promotes Efficient Clathrin Binding and Assembly

    PubMed Central

    Moshkanbaryans, Lia; Xue, Jing; Wark, Jesse Ray; Robinson, Phillip James

    2016-01-01

    The clathrin heavy chain N-terminal domain interacts with endocytic adapter proteins via clathrin binding motifs to assemble clathrin triskelia into cages. However, the precise mechanism of clathrin assembly is not yet known. Clathrin assembly protein AP180 has more clathrin binding motifs than any other endocytic protein and has a major role in the assembly of the clathrin coat during synaptic vesicle biogenesis. We now demonstrate that some of the previously identified binding motifs in AP180 may be non-functional and that a non-conventional clathrin binding sequence has a major influence on AP180 function. The related protein, clathrin assembly lymphoid myeloid leukemia protein (CALM), has fewer clathrin binding motifs and functions ubiquitously in clathrin-mediated endocytosis. The C-terminal ~16 kDa sub-domain in AP180, which has relatively high similarity with CALM, was shown in earlier work to have an unexplained role in clathrin binding. We identified the specific sequences in this sub-domain that bind to clathrin. Evidence for a role for these sequences in promoting clathrin binding was examined using in vitro and ex vivo experiments that compared the clathrin binding ability of site mutants with the wild type sequence. A sequence conserved in both AP180 and CALM (LDSSLA[S/N]LVGNLGI) was found to be the major interaction site and mutation caused a deficit in clathrin assembly, which is the first example of a mutation having this effect. In contrast, single or double mutation of DL(L/F) motifs in full length AP180 had no significant effect on clathrin binding, despite higher clathrin affinity for isolated peptides containing these motifs. We conclude that the novel clathrin interaction sites identified here in CALM and AP180 have a major role in how these proteins interface with clathrin. This work advances the case that AP180 and CALM are required to use a combination of standard clathrin N-terminal domain binding motifs and the sequence identified here

  2. A Novel Sequence in AP180 and CALM Promotes Efficient Clathrin Binding and Assembly.

    PubMed

    Moshkanbaryans, Lia; Xue, Jing; Wark, Jesse Ray; Robinson, Phillip James; Graham, Mark Evan

    2016-01-01

    The clathrin heavy chain N-terminal domain interacts with endocytic adapter proteins via clathrin binding motifs to assemble clathrin triskelia into cages. However, the precise mechanism of clathrin assembly is not yet known. Clathrin assembly protein AP180 has more clathrin binding motifs than any other endocytic protein and has a major role in the assembly of the clathrin coat during synaptic vesicle biogenesis. We now demonstrate that some of the previously identified binding motifs in AP180 may be non-functional and that a non-conventional clathrin binding sequence has a major influence on AP180 function. The related protein, clathrin assembly lymphoid myeloid leukemia protein (CALM), has fewer clathrin binding motifs and functions ubiquitously in clathrin-mediated endocytosis. The C-terminal ~16 kDa sub-domain in AP180, which has relatively high similarity with CALM, was shown in earlier work to have an unexplained role in clathrin binding. We identified the specific sequences in this sub-domain that bind to clathrin. Evidence for a role for these sequences in promoting clathrin binding was examined using in vitro and ex vivo experiments that compared the clathrin binding ability of site mutants with the wild type sequence. A sequence conserved in both AP180 and CALM (LDSSLA[S/N]LVGNLGI) was found to be the major interaction site and mutation caused a deficit in clathrin assembly, which is the first example of a mutation having this effect. In contrast, single or double mutation of DL(L/F) motifs in full length AP180 had no significant effect on clathrin binding, despite higher clathrin affinity for isolated peptides containing these motifs. We conclude that the novel clathrin interaction sites identified here in CALM and AP180 have a major role in how these proteins interface with clathrin. This work advances the case that AP180 and CALM are required to use a combination of standard clathrin N-terminal domain binding motifs and the sequence identified here

  3. From the dual function lead AP2238 to AP2469, a multi-target-directed ligand for the treatment of Alzheimer's disease

    PubMed Central

    Tarozzi, Andrea; Bartolini, Manuela; Piazzi, Lorna; Valgimigli, Luca; Amorati, Riccardo; Bolondi, Cecilia; Djemil, Alice; Mancini, Francesca; Andrisano, Vincenza; Rampa, Angela

    2014-01-01

    The development of drugs with different pharmacological properties appears to be an innovative therapeutic approach for Alzheimer's disease. In this article, we describe a simple structural modification of AP2238, a first dual function lead, in particular the introduction of the catechol moiety performed in order to search for multi-target ligands. The new compound AP2469 retains anti-acetylcholinesterase (AChE) and beta-site amyloid precursor protein cleaving enzyme (BACE)1 activities compared to the reference, and is also able to inhibit Aβ42 self-aggregation, Aβ42 oligomer-binding to cell membrane and subsequently reactive oxygen species formation in both neuronal and microglial cells. The ability of AP2469 to interfere with Aβ42 oligomer-binding to neuron and microglial cell membrane gives this molecule both neuroprotective and anti-inflammatory properties. These findings, together with its strong chain-breaking antioxidant performance, make AP2469 a potential drug able to modify the course of the disease. PMID:25505579

  4. The kinesin KIF16B mediates apical transcytosis of transferrin receptor in AP-1B-deficient epithelia

    PubMed Central

    Perez Bay, Andres E; Schreiner, Ryan; Mazzoni, Francesca; Carvajal-Gonzalez, Jose M; Gravotta, Diego; Perret, Emilie; Lehmann Mantaras, Gullermo; Zhu, Yuan-Shan; Rodriguez-Boulan, Enrique J

    2013-01-01

    Polarized epithelial cells take up nutrients from the blood through receptors that are endocytosed and recycle back to the basolateral plasma membrane (PM) utilizing the epithelial-specific clathrin adaptor AP-1B. Some native epithelia lack AP-1B and therefore recycle cognate basolateral receptors to the apical PM, where they carry out important functions for the host organ. Here, we report a novel transcytotic pathway employed by AP-1B-deficient epithelia to relocate AP-1B cargo, such as transferrin receptor (TfR), to the apical PM. Lack of AP-1B inhibited basolateral recycling of TfR from common recycling endosomes (CRE), the site of function of AP-1B, and promoted its transfer to apical recycling endosomes (ARE) mediated by the plus-end kinesin KIF16B and non-centrosomal microtubules, and its delivery to the apical membrane mediated by the small GTPase rab11a. Hence, our experiments suggest that the apical recycling pathway of epithelial cells is functionally equivalent to the rab11a-dependent TfR recycling pathway of non-polarized cells. They define a transcytotic pathway important for the physiology of native AP-1B-deficient epithelia and report the first microtubule motor involved in transcytosis. PMID:23749212

  5. 77 FR 24480 - Application for New Awards; Advanced Placement (AP) Test Fee Program-Reopening the AP Test Fee...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-24

    .... ACTION: Notice reopening the AP Test Fee fiscal year 2012 competition. Catalog of Federal Domestic Assistance (CFDA) Number: 84.330B. SUMMARY: On February 15, 2012, we published in the Federal Register (77 FR... Application for New Awards; Advanced Placement (AP) Test Fee Program--Reopening the AP Test Fee Fiscal...

  6. Thermodynamics of Damaged DNA Binding and Catalysis by Human AP Endonuclease 1

    PubMed Central

    Miroshnikova, A. D.; Kuznetsova, A. A.; Kuznetsov, N. A.; Fedorova, O. S.

    2016-01-01

    Apurinic/apyrimidinic (AP) endonucleases play an important role in DNA repair and initiation of AP site elimination. One of the most topical problems in the field of DNA repair is to understand the mechanism of the enzymatic process involving the human enzyme APE1 that provides recognition of AP sites and efficient cleavage of the 5’-phosphodiester bond. In this study, a thermodynamic analysis of the interaction between APE1 and a DNA substrate containing a stable AP site analog lacking the C1’ hydroxyl group (F site) was performed. Based on stopped-flow kinetic data at different temperatures, the steps of DNA binding, catalysis, and DNA product release were characterized. The changes in the standard Gibbs energy, enthalpy, and entropy of sequential specific steps of the repair process were determined. The thermodynamic analysis of the data suggests that the initial step of the DNA substrate binding includes formation of non-specific contacts between the enzyme binding surface and DNA, as well as insertion of the amino acid residues Arg177 and Met270 into the duplex, which results in the removal of “crystalline” water molecules from DNA grooves. The second binding step involves the F site flipping-out process and formation of specific contacts between the enzyme active site and the everted 5’-phosphate-2’-deoxyribose residue. It was shown that non-specific interactions between the binding surfaces of the enzyme and DNA provide the main contribution into the thermodynamic parameters of the DNA product release step. PMID:27099790

  7. AP600 containment purge radiological analysis

    SciTech Connect

    O`Connor, M.; Schulz, J.; Tan, C.

    1995-02-01

    The AP600 Project is a passive pressurized water reactor power plant which is part of the Design Certification and First-of-a-Kind Engineering effort under the Advanced Light Water Reactor program. Included in this process is the design of the containment air filtration system which will be the subject of this paper. We will compare the practice used by previous plants with the AP600 approach to meet the goals of industry standards in sizing the containment air filtration system. The radiological aspects of design are of primary significance and will be the focus of this paper. The AP600 Project optimized the design to combine the functions of the high volumetric flow rate, low volumetric flow rate, and containment cleanup and other filtration systems into one multi-functional system. This achieves a more simplified, standardized, and lower cost design. Studies were performed to determine the possible concentrations of radioactive material in the containment atmosphere and the effectiveness of the purge system to keep concentrations within 10CFR20 limits and within offsite dose objectives. The concentrations were determined for various reactor coolant system leakage rates and containment purge modes of operation. The resultant concentrations were used to determine the containment accessibility during various stages of normal plant operation including refueling. The results of the parametric studies indicate that a dual train purge system with a capacity of 4,000 cfm per train is more than adequate to control the airborne radioactivity levels inside containment during normal plant operation and refueling, and satisfies the goals of ANSI/ANS-56.6-1986 and limits the amount of radioactive material released to the environment per ANSI/ANS 59.2-1985 to provide a safe environment for plant personnel and offsite residents.

  8. Ozone mitigation tests at the APS

    SciTech Connect

    Kuzay, T.M.; Collins, J.T.; Pisharody, M.; Job, P.K.; Wang Zhibi

    1996-09-01

    Ozone is generated in the APS experimental stations whenever the x-ray beam has a chance to interact with air. Ozone concentrations in an experimental station have to be below a certain defined limit (current OSHA regulations specify 0.08 ppm as the maximum limit) before an experimenter can reenter the hutch. This limit is said to be currently under study for a downward adjustment. One method of depleting the ozone generated in an experimental station is mitigation through either adsorption or direct destruction. In recent tests, both methods were tried using commercially available units. Test results and some analytical predictions are presented.

  9. Correction magnet power supplies for APS machine

    SciTech Connect

    Kang, Y.G.

    1991-04-01

    A number of correction magnets are required for the advanced photon source (APS) machine to correct the beam. There are five kinds of correction magnets for the storage ring, two for the injector synchrotron, and two for the positron accumulator ring (PAR). Table I shoes a summary of the correction magnet power supplies for the APS machine. For the storage ring, the displacement of the quadrupole magnets due to the low frequency vibration below 25 Hz has the most significant effect on the stability of the positron closed orbit. The primary external source of the low frequency vibration is the ground motion of approximately 20 {mu}m amplitude, with frequency components concentrated below 10 Hz. These low frequency vibrations can be corrected by using the correction magnets, whose field strengths are controlled individually through the feedback loop comprising the beam position monitoring system. The correction field require could be either positive or negative. Thus for all the correction magnets, bipolar power supplies (BPSs) are required to produce both polarities of correction fields. Three different types of BPS are used for all the correction magnets. Type I BPSs cover all the correction magnets for the storage ring, except for the trim dipoles. The maximum output current of the Type I BPS is 140 Adc. A Type II BPS powers a trim dipole, and its maximum output current is 60 Adc. The injector synchrotron and PAR correction magnets are powered form Type III BPSs, whose maximum output current is 25 Adc.

  10. The APS control system network upgrade.

    SciTech Connect

    Sidorowicz, K. v.; Leibfritz, D.; McDowell, W. P.

    1999-10-22

    When it was installed,the Advanced Photon Source (APS) control system network was at the state-of-the-art. Different aspects of the system have been reported at previous meetings [1,2]. As loads on the controls network have increased due to newer and faster workstations and front-end computers, we have found performance of the system declining and have implemented an upgraded network. There have been dramatic advances in networking hardware in the last several years. The upgraded APS controls network replaces the original FDDI backbone and shared Ethernet hubs with redundant gigabit uplinks and fully switched 10/100 Ethernet switches with backplane fabrics in excess of 20 Gbits/s (Gbps). The central collapsed backbone FDDI concentrator has been replaced with a Gigabit Ethernet switch with greater than 30 Gbps backplane fabric. Full redundancy of the system has been maintained. This paper will discuss this upgrade and include performance data and performance comparisons with the original network.

  11. AP1000{sup R} nuclear power plant safety overview for spent fuel cooling

    SciTech Connect

    Gorgemans, J.; Mulhollem, L.; Glavin, J.; Pfister, A.; Conway, L.; Schulz, T.; Oriani, L.; Cummins, E.; Winters, J.

    2012-07-01

    The AP1000{sup R} plant is an 1100-MWe class pressurized water reactor with passive safety features and extensive plant simplifications that enhance construction, operation, maintenance, safety and costs. The AP1000 design uses passive features to mitigate design basis accidents. The passive safety systems are designed to function without safety-grade support systems such as AC power, component cooling water, service water or HVAC. Furthermore, these passive features 'fail safe' during a non-LOCA event such that DC power and instrumentation are not required. The AP1000 also has simple, active, defense-in-depth systems to support normal plant operations. These active systems provide the first level of defense against more probable events and they provide investment protection, reduce the demands on the passive features and support the probabilistic risk assessment. The AP1000 passive safety approach allows the plant to achieve and maintain safe shutdown in case of an accident for 72 hours without operator action, meeting the expectations provided in the U.S. Utility Requirement Document and the European Utility Requirements for passive plants. Limited operator actions are required to maintain safe conditions in the spent fuel pool via passive means. In line with the AP1000 approach to safety described above, the AP1000 plant design features multiple, diverse lines of defense to ensure spent fuel cooling can be maintained for design-basis events and beyond design-basis accidents. During normal and abnormal conditions, defense-in-depth and other systems provide highly reliable spent fuel pool cooling. They rely on off-site AC power or the on-site standby diesel generators. For unlikely design basis events with an extended loss of AC power (i.e., station blackout) or loss of heat sink or both, spent fuel cooling can still be provided indefinitely: - Passive systems, requiring minimal or no operator actions, are sufficient for at least 72 hours under all possible pool

  12. EPA’s AP-42 development methodology: Converting or rerating current AP-42 datasets

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In August 2013, the U.S. Environmental Protection Agency’s (EPA) published their new methodology for updating the Compilation of Air Pollution Emission Factors (AP-42). The “Recommended Procedures for Development of Emissions Factors and Use of the WebFIRE Database” instructs that the ratings of the...

  13. Evaluating EPA’s AP-42 development methodology to convert or rerate current AP-42 datasets

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In August 2013, the U.S. Environmental Protection Agency’s (EPA) published their new methodology for updating the Compilation of Air Pollution Emission Factors (AP-42). The “Recommended Procedures for Development of Emissions Factors and Use of the WebFIRE Database” instructs that the ratings of the...

  14. Transcriptomic analysis by RNA-seq reveals AP-1 pathway as key regulator that green tea may rely on to inhibit lung tumorigenesis.

    PubMed

    Pan, Jing; Zhang, Qi; Xiong, Donghai; Vedell, Peter; Yan, Ying; Jiang, Hui; Cui, Peng; Ding, Feng; Tichelaar, Jay W; Wang, Yian; Lubet, Ronald A; You, Ming

    2014-01-01

    Green tea is a promising chemopreventive agent for lung cancer. Multiple signaling events have been reported, however, the relative importance of these mechanisms in mediating the chemopreventive function of green tea is unclear. In the present study, to examine the involvement of AP-1 in green tea polyphenols induced tumor inhibition, human NSCLC cell line H1299 and mouse SPON 10 cells were identified as AP-1 dependent, as these two lines exhibit high constitutive AP-1 activity, and when TAM67 expression was induced with doxycycline, cell growth was inhibited and correlated with suppressed AP-1 activity. RNA-seq was used to determine the global transcriptional effects of AP-1 inhibition and also uncover the possible involvement of AP-1 in tea polyphenols induced chemoprevention. TAM67 mediated changes in gene expression were identified, and within down-regulated genes, AP-1 was identified as a key transcription regulator. RNA-seq analysis revealed that Polyphenon E-treated cells shared 293 commonly down-regulated genes within TAM67 expressing H1299 cells, and by analysis of limited Chip-seq data, over 10% of the down-regulated genes contain a direct AP-1 binding site, indicating that Polyphenon E elicits chemopreventive activity by regulating AP-1 target genes. Conditional TAM67 expressing transgenic mice and NSCLC cell lines were used to further confirm that the chemopreventive activity of green tea is AP-1 dependent. Polyphenon E lost its chempreventive function both in vitro and in vivo when AP-1 was inhibited, indicating that AP-1 inhibition is a major pathway through which green tea exhibits chemopreventive effects. PMID:24343902

  15. The high-overtone p-mode spectrum of the rapidly oscillating Ap star HR 1217 (HD 24712) - Results of a frequency analysis of 324 hr of multi-site photometric observations obtained during a 46-d time-span in 1986

    NASA Technical Reports Server (NTRS)

    Kurtz, D. W.; Martinez, P.; Seeman, J.; Matthews, J. M.; Cropper, Mark

    1989-01-01

    The present high-speed photometry for the rapidly oscillating Ap star HR 1217 indicates that the 6-min light variations are amplitude-modulated, in phase with the magnetic variations. Amplitude and magnetic maxima coincide, while the mean light minimum occurs at a perhaps significantly different time. It is established that HR 1217 is an oblique rotator with a centered dipole magnetic field, as well as an oblique pulsator whose pulsation and magnetic axes are aligned. There are six principal frequencies of pulsation; the secondary frequencies indicate that the principal ones are amplitude-modulated on a time-scale of months.

  16. Distinct catalytic activity and in vivo roles of the ExoIII and EndoIV AP endonucleases from Sulfolobus islandicus.

    PubMed

    Yan, Zhou; Huang, Qihong; Ni, Jinfeng; Shen, Yulong

    2016-09-01

    AP endonuclease cleaves the phosphodiester bond 5'- to the AP (apurinic or apyrimidinic) sites and is one of the major enzymes involved in base excision repair. So far, the properties of several archaeal AP endonuclease homologues have been characterized in vitro, but little is known about their functions in vivo. Herein, we report on the biochemical and genetic analysis of two AP endonucleases, SisExoIII and SisEndoIV, from the hyperthermophilic crenarchaeon Sulfolobus islandicus REY15A. Both SisExoIII and SisEndoIV exhibit AP endonuclease activity, but neither of them has 3'-5' exonuclease activity. SisExoIII and SisEndoIV have similar K M values on the substrate containing an AP site, but the latter cleaves the AP substrate at a dramatically higher catalytic rate than the former. Unlike other AP endonucleases identified in archaea, SisExoIII and SisEndoIV do not exhibit any cleavage activity on DNA having oxidative damage (8-oxo-dG) or uracil. Genetic analysis revealed that neither gene is essential for cell viability, and the growth of ∆SiRe_2666 (endoIV), ∆SiRe_0100 (exoIII), and ∆SiRe_0100∆SiRe_2666 is not affected under normal growth conditions. However, ∆SiRe_2666 exhibits higher sensitivity to the alkylating agent methyl methanesulfonate (MMS) than ∆SiRe_0100. Over-expression of SiRe_0100 can partially complement the sensitivity of ∆SiRe_2666 to MMS, suggesting a backup role of SisExoIII in AP site processing in vivo. Intriguingly, over-expression of SisEndoIV renders the strain more sensitive to MMS than the control. Taken together, we conclude that SisEndoIV, but not SisExoIII, is the main AP endonuclease that participates directly in base excision repair in S. islandicus. PMID:27457080

  17. Vibration study of the APS magnet support assemblies

    SciTech Connect

    Wambsganss, M.W.; Jendrzejczyk, J.A.; Chen, S.S.

    1990-11-01

    Stability of the positron closed orbit is a requirement for successful operation of the Advanced Photon Source. The fact that vibration of the storage ring quadrupole magnets can lead to distortion of the positron closed orbit and to potentially unacceptable beam emittance growth provides the motivation for the subject studies. Low frequency vibrations can be controlled with steering magnets using feedback systems, provided the vibration amplitudes are within the dynamic range of the controllers. High frequency vibration amplitudes, on the other hand, are out of the range of the controller and, therefore must be limited to ensure the emittance growth will not exceed a prescribed value. Vibration criteria were developed based on the requirement that emittance growth be limited to 10 percent. Recognizing that the quadrupole magnets have the most significant effect, three different scenarios were considered: vibration of a single quadrupole within the storage ring, random vibration of all the quadrupoles in the ring, and the hypothetical case of a plane wave sweeping across the site and the quadrupoles following the motion of the plane wave. The maximum allowable peak vibration amplitudes corresponding to these three vibration scenarios are given. The criteria associated with the passage of a plane wave is dependent on wavelength, or, alternatively, on frequency given the wave speed. The wave speed used is that measured as a part of the geotechnical investigation at the APS site.

  18. Structure-guided creation of AcAP5-derived and platelet targeted factor Xa inhibitors.

    PubMed

    Zhu, Yuanjun; Lin, Yuan; Liu, Aihua; Shui, Mengyang; Li, Ruyi; Liu, Xiaoyan; Hu, Wenhui; Wang, Yinye

    2015-06-15

    Anticoagulants and anti-platelet agents are simultaneously administrated in clinical practice (i.e. percutaneous coronary intervention), which cause significant risk of systemic bleeding. Targeted delivery of anticoagulants to the activated platelets at sites of vascular injuries may condense the site-specific anticoagulant effect and reduce the hemorrhage side effects in uninjured vessels. To this end, we prepared three ancylostoma caninum anticoagulant peptide 5 (AcAP5) variants NR1, NR2 and NR3 engineered with a platelet-binding Arg-Gly-Asp (RGD) motif and evaluated their anti-Factor Xa (FXa) and platelet-binding effects. These RGD-containing AcAP5 variants were capable of interacting with platelet receptor αIIbβ3 as shown in computational analysis. All variants, especially NR2 and NR3, retained entirely the anti-FXa function of parent AcAP5. Moreover, they prevented the formation of occlusive thrombi in rat carotid artery injury model, suggesting that they inhibit platelet aggregation in vivo. Further functional investigation of NR3 demonstrated that NR3 inhibited platelet aggregation in vitro and FXa activity in vivo, and prolonged the coagulation time, all in a dose-dependent manner. Through flow cytometry assay, we confirmed the binding of NR3 to αIIbβ3 receptor. In mouse model of carotid artery endothelium injury, NR3-treated mice showed less tail bleeding time than AcAP5-treated mice, and aspirin plus NR3 treatment exhibited moderate reduction of blood loss compared with aspirin plus AcAP5 treatment. These results indicate the feasibility to engineer a novel FXa inhibitor specifically targeting the activated platelets, which centralizes its anticoagulation efficacy in the injured vascular endothelium and reduces the risk of systemic bleeding. PMID:25887920

  19. A single β adaptin contributes to AP1 and AP2 complexes and clathrin function in Dictyostelium.

    PubMed

    Sosa, R Thomas; Weber, Michelle M; Wen, Yujia; O'Halloran, Theresa J

    2012-02-01

    The assembly of clathrin-coated vesicles is important for numerous cellular processes, including nutrient uptake and membrane organization. Important contributors to clathrin assembly are four tetrameric assembly proteins, also called adaptor proteins (APs), each of which contains a β subunit. We identified a single β subunit, named β1/2, that contributes to both the AP1 and AP2 complexes of Dictyostelium. Disruption of the gene encoding β1/2 resulted in severe defects in growth, cytokinesis and development. Additionally, cells lacking β1/2 displayed profound osmoregulatory defects including the absence of contractile vacuoles and mislocalization of contractile vacuole markers. The phenotypes of β1/2 null cells were most similar to previously described phenotypes of clathrin and AP1 mutants, supporting a particularly important contribution of AP1 to clathrin pathways in Dictyostelium cells. The absence of β1/2 in cells led to significant reductions in the protein amounts of the medium-sized subunits of the AP1 and AP2 complexes, establishing a role for the β subunit in the stability of the medium subunits. Dictyostelium β1/2 could resemble a common ancestor of the more specialized β1 and β2 subunits of the vertebrate AP complexes. Our results support the essential contribution of a single β subunit to the stability and function of AP1 and AP2 in a simple eukaryote. PMID:22050483

  20. Radiation characteristics of scintillator coupled CMOS APS for radiography conditions

    NASA Astrophysics Data System (ADS)

    Kim, Kwang Hyun; Kim, Soongpyung; Kang, Dong-Won; Kim, Dong-Kie

    2006-11-01

    Under industrial radiography conditions, we analyzed short-term radiation characteristics of scintillator coupled CMOS APS (hereinafter SC CMOS APS). By means of experimentation, the contribution of the transmitted X-ray through the scintillator to the properties of the CMOS APS and the afterimage, generated in the acquired image even at low dose condition, were investigated. To see the transmitted X-ray effects on the CMOS APS, Fein focus™ X-ray machine, two scintillators of Lanex™ Fine and Regular, and two CMOS APS array of RadEye™ were used under the conditions of 50 kV p/1 mAs and 100 kV p/1 mAs. By measuring the transmitted X-ray on signal and Noise Power Spectrum, we analytically examined the generation mechanism of the afterimage, based on dark signal or dark current increase in the sensor, and explained the afterimage in the SC CMOS APS.

  1. Design soil profiles for seismic analyses of AP600 plant standard design

    SciTech Connect

    Ostadan, F.; Gross, K.K.; Liu, C.I.T.; Orr, R.S.

    1996-12-01

    Future nuclear power plants are based on standard designs. For seismic qualification, a variety of subsurface conditions are considered in the design. While the soil properties play a significant role in the seismic responses, an unlimited number of site conditions can be postulated for analyses. In this paper, a systematic and effective approach is described to arrive at the design soil profiles for the AP600 nuclear plant.

  2. The AP-2 Adaptor β2 Appendage Scaffolds Alternate Cargo Endocytosis

    PubMed Central

    Keyel, Peter A.; Thieman, James R.; Roth, Robyn; Erkan, Elif; Everett, Eric T.; Watkins, Simon C.; Heuser, John E.

    2008-01-01

    The independently folded appendages of the large α and β2 subunits of the endocytic adaptor protein (AP)-2 complex coordinate proper assembly and operation of endocytic components during clathrin-mediated endocytosis. The β2 subunit appendage contains a common binding site for β-arrestin or the autosomal recessive hypercholesterolemia (ARH) protein. To determine the importance of this interaction surface in living cells, we used small interfering RNA-based gene silencing. The effect of extinguishing β2 subunit expression on the internalization of transferrin is considerably weaker than an AP-2 α subunit knockdown. We show the mild sorting defect is due to fortuitous substitution of the β2 chain with the closely related endogenous β1 subunit of the AP-1 adaptor complex. Simultaneous silencing of both β1 and β2 subunit transcripts recapitulates the strong α subunit RNA interference (RNAi) phenotype and results in loss of ARH from endocytic clathrin coats. An RNAi-insensitive β2-yellow fluorescent protein (YFP) expressed in the β1 + β2-silenced background restores cellular AP-2 levels, robust transferrin internalization, and ARH colocalization with cell surface clathrin. The importance of the β appendage platform subdomain over clathrin for precise deposition of ARH at clathrin assembly zones is revealed by a β2-YFP with a disrupted ARH binding interface, which does not restore ARH colocalization with clathrin. We also show a β-arrestin 1 mutant, which engages coated structures in the absence of any G protein-coupled receptor stimulation, colocalizes with β2-YFP and clathrin even in the absence of an operational clathrin binding sequence. These findings argue against ARH and β-arrestin binding to a site upon the β2 appendage platform that is later obstructed by polymerized clathrin. We conclude that ARH and β-arrestin depend on a privileged β2 appendage site for proper cargo recruitment to clathrin bud sites. PMID:18843039

  3. Control units for APS power supplies

    SciTech Connect

    Despe, O.D.; Saunders, C.; McGhee, D.G.

    1993-07-01

    The Advanced Photon Source (APS) accelerator facility is made up of five major subsystems in addition to the linac: the positron accumulator ring (PAR), low energy transport (LET), booster synchrotron (SYNCH), high energy transport (HET), the storage ring (SR). Each subsystem has multiple magnet power supply combinations, some requiring multiple of operation. These magnet and power supply combinations computer controlled and monitored. The power supply control unit (PSCU) is the first layer of hardware and software above the power supply itself and is described in this paper. The description includes the basic philosophy for each of operation and how it influences the topology and of implementing control. The design of the analog reference blocks (ARBs) influenced the design of other custom functions well as the feedback controls for vibration and other dynamic corrections. The command set supported by the PSCU is discussed.

  4. Endonuclease IV Is the major apurinic/apyrimidinic endonuclease in Mycobacterium tuberculosis and is important for protection against oxidative damage.

    PubMed

    Puri, Rupangi Verma; Singh, Nisha; Gupta, Rakesh K; Tyagi, Anil K

    2013-01-01

    During the establishment of an infection, bacterial pathogens encounter oxidative stress resulting in the production of DNA lesions. Majority of these lesions are repaired by base excision repair (BER) pathway. Amongst these, abasic sites are the most frequent lesions in DNA. Class II apurinic/apyrimidinic (AP) endonucleases play a major role in BER of damaged DNA comprising of abasic sites. Mycobacterium tuberculosis, a deadly pathogen, resides in the human macrophages and is continually subjected to oxidative assaults. We have characterized for the first time two AP endonucleases namely Endonuclease IV (End) and Exonuclease III (XthA) that perform distinct functions in M.tuberculosis. We demonstrate that M.tuberculosis End is a typical AP endonuclease while XthA is predominantly a 3'→5' exonuclease. The AP endonuclease activity of End and XthA was stimulated by Mg(2+) and Ca(2+) and displayed a preferential recognition for abasic site paired opposite to a cytosine residue in DNA. Moreover, End exhibited metal ion independent 3'→5' exonuclease activity while in the case of XthA this activity was metal ion dependent. We demonstrate that End is not only a more efficient AP endonuclease than XthA but it also represents the major AP endonuclease activity in M.tuberculosis and plays a crucial role in defense against oxidative stress. PMID:23936515

  5. Matrix Proteins of Nipah and Hendra Viruses Interact with Beta Subunits of AP-3 Complexes

    PubMed Central

    Sun, Weina; McCrory, Thomas S.; Khaw, Wei Young; Petzing, Stephanie; Myers, Terrell

    2014-01-01

    ABSTRACT Paramyxoviruses and other negative-strand RNA viruses encode matrix proteins that coordinate the virus assembly process. The matrix proteins link the viral glycoproteins and the viral ribonucleoproteins at virus assembly sites and often recruit host machinery that facilitates the budding process. Using a co-affinity purification strategy, we have identified the beta subunit of the AP-3 adapter protein complex, AP3B1, as a binding partner for the M proteins of the zoonotic paramyxoviruses Nipah virus and Hendra virus. Binding function was localized to the serine-rich and acidic Hinge domain of AP3B1, and a 29-amino-acid Hinge-derived polypeptide was sufficient for M protein binding in coimmunoprecipitation assays. Virus-like particle (VLP) production assays were used to assess the relationship between AP3B1 binding and M protein function. We found that for both Nipah virus and Hendra virus, M protein expression in the absence of any other viral proteins led to the efficient production of VLPs in transfected cells, and this VLP production was potently inhibited upon overexpression of short M-binding polypeptides derived from the Hinge region of AP3B1. Both human and bat (Pteropus alecto) AP3B1-derived polypeptides were highly effective at inhibiting the production of VLPs. VLP production was also impaired through small interfering RNA (siRNA)-mediated depletion of AP3B1 from cells. These findings suggest that AP-3-directed trafficking processes are important for henipavirus particle production and identify a new host protein-virus protein binding interface that could become a useful target in future efforts to develop small molecule inhibitors to combat paramyxoviral infections. IMPORTANCE Henipaviruses cause deadly infections in humans, with a mortality rate of about 40%. Hendra virus outbreaks in Australia, all involving horses and some involving transmission to humans, have been a continuing problem. Nipah virus caused a large outbreak in Malaysia in 1998

  6. Low Earth orbit assessment of proton anisotropy using AP8 and AP9 trapped proton models

    NASA Astrophysics Data System (ADS)

    Badavi, Francis F.; Walker, Steven A.; Santos Koos, Lindsey M.

    2015-04-01

    The completion of the International Space Station (ISS) in 2011 has provided the space research community with an ideal evaluation and testing facility for future long duration human activities in space. Ionized and secondary neutral particles radiation measurements inside ISS form the ideal tool for validation of radiation environmental models, nuclear reaction cross sections and transport codes. Studies using thermo-luminescent detectors (TLD), tissue equivalent proportional counter (TPEC), and computer aided design (CAD) models of early ISS configurations confirmed that, as input, computational dosimetry at low Earth orbit (LEO) requires an environmental model with directional (anisotropic) capability to properly describe the exposure of trapped protons within ISS. At LEO, ISS encounters exposure from trapped electrons, protons and geomagnetically attenuated galactic cosmic rays (GCR). For short duration studies at LEO, one can ignore trapped electrons and ever present GCR exposure contributions during quiet times. However, within the trapped proton field, a challenge arises from properly estimating the amount of proton exposure acquired. There exist a number of models to define the intensity of trapped particles. Among the established trapped models are the historic AE8/AP8, dating back to the 1980s and the recently released AE9/AP9/SPM. Since at LEO electrons have minimal exposure contribution to ISS, this work ignores the AE8 and AE9 components of the models and couples a measurement derived anisotropic trapped proton formalism to omnidirectional output from the AP8 and AP9 models, allowing the assessment of the differences between the two proton models. The assessment is done at a target point within the ISS-11A configuration (circa 2003) crew quarter (CQ) of Russian Zvezda service module (SM), during its ascending and descending nodes passes through the south Atlantic anomaly (SAA). The anisotropic formalism incorporates the contributions of proton narrow

  7. Low Earth orbit assessment of proton anisotropy using AP8 and AP9 trapped proton models.

    PubMed

    Badavi, Francis F; Walker, Steven A; Santos Koos, Lindsey M

    2015-04-01

    The completion of the International Space Station (ISS) in 2011 has provided the space research community with an ideal evaluation and testing facility for future long duration human activities in space. Ionized and secondary neutral particles radiation measurements inside ISS form the ideal tool for validation of radiation environmental models, nuclear reaction cross sections and transport codes. Studies using thermo-luminescent detectors (TLD), tissue equivalent proportional counter (TPEC), and computer aided design (CAD) models of early ISS configurations confirmed that, as input, computational dosimetry at low Earth orbit (LEO) requires an environmental model with directional (anisotropic) capability to properly describe the exposure of trapped protons within ISS. At LEO, ISS encounters exposure from trapped electrons, protons and geomagnetically attenuated galactic cosmic rays (GCR). For short duration studies at LEO, one can ignore trapped electrons and ever present GCR exposure contributions during quiet times. However, within the trapped proton field, a challenge arises from properly estimating the amount of proton exposure acquired. There exist a number of models to define the intensity of trapped particles. Among the established trapped models are the historic AE8/AP8, dating back to the 1980s and the recently released AE9/AP9/SPM. Since at LEO electrons have minimal exposure contribution to ISS, this work ignores the AE8 and AE9 components of the models and couples a measurement derived anisotropic trapped proton formalism to omnidirectional output from the AP8 and AP9 models, allowing the assessment of the differences between the two proton models. The assessment is done at a target point within the ISS-11A configuration (circa 2003) crew quarter (CQ) of Russian Zvezda service module (SM), during its ascending and descending nodes passes through the south Atlantic anomaly (SAA). The anisotropic formalism incorporates the contributions of proton narrow

  8. DNA conformation driven by AP-1 triggers cell-specific expression via a strong epithelial enhancer.

    PubMed

    Virolle, T; Djabari, Z; Ortonne, J P; Aberdam, D

    2000-10-01

    We report here the characterization of the regulatory region of the human LAMA3 gene, coding for the alpha3A chain of laminin-5. A 202 bp fragment is sufficient to confer epithelial-specific expression to a thymidine kinase promoter through the cooperative effect of three AP-1 binding sites. Remarkably, removal of the sequences located between the AP-1 sites does not modify the promoter activity in keratinocytes but allows strong expression in fibroblasts. Replacement of the deleted sequences by non-homologous ones fully restores the restricted enhancement in keratinocytes. Functional analysis and mutagenesis experiments demonstrate that a minimal distance between the AP-1 sites is required for the enhancer DNA fragment to adopt a particular conformation driven by the binding of Jun-Fos heterodimers. In non-permissive cells, this conformation leads to the anchorage of non-DNA-binding fibroblastic cofactors to form an inhibitory ternary complex. Therefore, our results describe for the first time an unusual conformation-dependent epithelial-specific enhancer. PMID:11269498

  9. Terminal deoxynucleotidyl transferase is down-regulated by AP-1-like regulatory elements in human lymphoid cells

    PubMed Central

    Peralta-Zaragoza, Oscar; Recillas-Targa, Félix; Madrid-Marina, Vicente

    2004-01-01

    Terminal deoxynucleotidyl transferase (TdT) is a template-independent DNA polymerase that catalyses the incorporation of deoxyribonucleotides into the 3′-hydroxyl end of DNA templates and is thought to increase junctional diversity of antigen receptor genes. TdT is expressed only on immature lymphocytes and acute lymphoblastic leukaemia cells and its transcriptional expression is tightly regulated. We had previously found that protein kinase C (PKC) activation down-regulates TdT expression. PKC-activation induces the synthesis of the Fos and Jun proteins, known as the major components of activation protein 1 (AP-1) transcriptional factor implicated in transcriptional control. Here we report the identification of several DNA–protein interactions within the TdT promoter region in non-TdT expressing human cells. Sequence analysis revealed the presence of a putative AP-1-like DNA-binding site, suggesting that AP-1 may play a relevant role in TdT transcriptional regulation. Using a different source of nuclear extracts and the AP-1–TdT motif as a probe we identified several DNA-protein retarded complexes in electrophoretic mobility shift assays. Super-band shifting analysis using an antibody against c-Jun protein confirmed that the main interaction is produced by a nuclear factor that belongs to the AP-1 family transcription factors. Our findings suggest that the TdT gene expression is down-regulated, at least in part, through AP-1-like transcription factors. PMID:15027905

  10. Terminal deoxynucleotidyl transferase is down-regulated by AP-1-like regulatory elements in human lymphoid cells.

    PubMed

    Peralta-Zaragoza, Oscar; Recillas-Targa, Félix; Madrid-Marina, Vicente

    2004-02-01

    Terminal deoxynucleotidyl transferase (TdT) is a template-independent DNA polymerase that catalyses the incorporation of deoxyribonucleotides into the 3'-hydroxyl end of DNA templates and is thought to increase junctional diversity of antigen receptor genes. TdT is expressed only on immature lymphocytes and acute lymphoblastic leukaemia cells and its transcriptional expression is tightly regulated. We had previously found that protein kinase C (PKC) activation down-regulates TdT expression. PKC-activation induces the synthesis of the Fos and Jun proteins, known as the major components of activation protein 1 (AP-1) transcriptional factor implicated in transcriptional control. Here we report the identification of several DNA-protein interactions within the TdT promoter region in non-TdT expressing human cells. Sequence analysis revealed the presence of a putative AP-1-like DNA-binding site, suggesting that AP-1 may play a relevant role in TdT transcriptional regulation. Using a different source of nuclear extracts and the AP-1-TdT motif as a probe we identified several DNA-protein retarded complexes in electrophoretic mobility shift assays. Super-band shifting analysis using an antibody against c-Jun protein confirmed that the main interaction is produced by a nuclear factor that belongs to the AP-1 family transcription factors. Our findings suggest that the TdT gene expression is down-regulated, at least in part, through AP-1-like transcription factors. PMID:15027905

  11. Opposing Effects of Zac1 and Curcumin on AP-1-Regulated Expressions of S100A7

    PubMed Central

    Chu, Yu-Wen; Liu, Shu-Ting; Cheng, Hsiao-Chun; Huang, Shih-Ming; Chang, Yung-Lung; Chiang, Chien-Ping; Liu, Ying-Chun; Wang, Wei-Ming

    2015-01-01

    ZAC, an encoding gene mapped at chromosome 6q24-q25 within PSORS1, was previously found over-expressed in the lower compartment of the hyperplastic epidermis in psoriatic lesions. Cytokines produced in the inflammatory dermatoses may drive AP-1 transcription factor to induce responsive gene expressions. We demonstrated that mZac1 can enhance AP-1-responsive S100A7 expression of which the encoding gene was located in PSORS4 with HaCaT keratinocytes. However, the mZac1-enhanced AP-1 transcriptional activity was suppressed by curcumin, indicating the anti-inflammatory property of this botanical agent and is exhibited by blocking the AP-1-mediated cross-talk between PSORS1 and PSORS4. Two putative AP-1-binding sites were found and demonstrated to be functionally important in the regulation of S100A7 promoter activity. Moreover, we found curcumin reduced the DNA-binding activity of AP-1 to the recognition element located in the S100A7 promoter. The S100A7 expression was found to be upregulated in the lesioned epidermis of atopic dermatitis and psoriasis, which is where this keratinocyte-derived chemoattractant engaged in the pro-inflammatory feedback loop. Understanding the regulatory mechanism of S100A7 expression will be helpful to develop therapeutic strategies for chronic inflammatory dermatoses via blocking the reciprocal stimuli between the inflammatory cells and keratinocytes. PMID:26633653

  12. Anti-cancer effect of snake venom toxin through down regulation of AP-1 mediated PRDX6 expression

    PubMed Central

    Son, Dong Ju; Song, Ho Sub; Kim, Jung Hyun; Ko, Seong Cheol; Song, Min Jong; Lee, Won Hyoung; Yoon, Joo Hee; Ham, Young Wan; Han, Sang Bae; Hong, Jin Tae

    2015-01-01

    Snake venom toxin (SVT) from Vipera lebetina turanica contains a mixture of different enzymes and proteins. Peroxiredoxin 6 (PRDX6) is known to be a stimulator of lung cancer cell growth. PRDX6 is a member of peroxidases, and has calcium-independent phospholipase A2 (iPLA2) activities. PRDX6 has an AP-1 binding site in its promoter region of the gene. Since AP-1 is implicated in tumor growth and PRDX6 expression, in the present study, we investigated whether SVT inhibits PRDX6, thereby preventing human lung cancer cell growth (A549 and NCI-H460) through inactivation of AP-1. A docking model study and pull down assay showed that SVT completely fits on the basic leucine zipper (bZIP) region of c-Fos of AP-1. SVT (0–10 μg/ml) inhibited lung cancer cell growth in a concentration dependent manner through induction of apoptotic cell death accompanied by induction of cleaved caspase-3, -8, -9, Bax, p21 and p53, but decreased cIAP and Bcl2 expression via inactivation of AP-1. In an xenograft in vivo model, SVT (0.5 mg/kg and 1 mg/kg) also inhibited tumor growth accompanied with the reduction of PRDX6 expression, but increased expression of proapoptotic proteins. These data indicate that SVT inhibits tumor growth via inhibition of PRDX6 activity through interaction with its transcription factor AP-1. PMID:26061816

  13. Project W-211, initial tank retrieval systems, description of operations for 241-AP-102 and 241-AP-104

    SciTech Connect

    RIECK, C.A.

    1999-02-25

    The primary purpose of the Initial Tank Retrieval Systems (ITRS) is to provide systems for retrieval of radioactive wastes stored in underground double-shell tanks (DSTS) for transfer to alternate storage, evaporation, pretreatment or treatment, while concurrently reducing risks associated with safety watch list and other DSTs. This Description of Operations (DOO) defines the control philosophy for the waste retrieval system for tanks 241-AP-102 (AP-102) and 241-AP-104 (AP-104). This DOO will provide a basis for the detailed design of the Retrieval Control System (RCS) for AP-102 and AP-104 and establishes test criteria for the RCS. The test criteria will be used during qualification testing and acceptance testing to verify operability.

  14. Sip1, a Conserved AP-1 Accessory Protein, Is Important for Golgi/Endosome Trafficking in Fission Yeast

    PubMed Central

    Yu, Yang; Kita, Ayako; Udo, Masako; Katayama, Yuta; Shintani, Mami; Park, Kwihwa; Hagihara, Kanako; Umeda, Nanae; Sugiura, Reiko

    2012-01-01

    We had previously identified the mutant allele of apm1+ that encodes a homolog of the mammalian μ 1A subunit of the clathrin-associated adaptor protein-1 (AP-1) complex and demonstrated that the AP-1 complex plays a role in Golgi/endosome trafficking, secretion, and vacuole fusion in fission yeast. Here, we isolated a mutant allele of its4+/sip1+, which encodes a conserved AP-1 accessory protein. The its4-1/sip1-i4 mutants and apm1-deletion cells exhibited similar phenotypes, including sensitivity to the calcineurin inhibitor FK506, Cl− and valproic acid as well as various defects in Golgi/endosomal trafficking and cytokinesis. Electron micrographs of sip1-i4 mutants revealed vacuole fragmentation and accumulation of abnormal Golgi-like structures and secretory vesicles. Overexpression of Apm1 suppressed defective membrane trafficking in sip1-i4 mutants. The Sip1-green fluorescent protein (GFP) co-localized with Apm1-mCherry at Golgi/endosomes, and Sip1 physically interacted with each subunit of the AP-1 complex. We found that Sip1 was a Golgi/endosomal protein and the sip1-i4 mutation affected AP-1 localization at Golgi/endosomes, thus indicating that Sip1 recruited the AP-1 complex to endosomal membranes by physically interacting with each subunit of this complex. Furthermore, Sip1 is required for the correct localization of Bgs1/Cps1, 1,3-β-D-glucan synthase to polarized growth sites. Consistently, the sip1-i4 mutants displayed a severe sensitivity to micafungin, a potent inhibitor of 1,3-β-D-glucan synthase. Taken together, our findings reveal a role for Sip1 in the regulation of Golgi/endosome trafficking in coordination with the AP-1 complex, and identified Bgs1, required for cell wall synthesis, as the new cargo of AP-1-dependent trafficking. PMID:23028933

  15. The AP2-like gene NsAP2 from water lily is involved in floral organogenesis and plant height.

    PubMed

    Luo, Huolin; Chen, Sumei; Jiang, Jiafu; Teng, Nianjun; Chen, Yu; Chen, Fadi

    2012-07-01

    APETALA2 (AP2) genes are ancient and widely distributed among the seed plants, and play an important role during the plant life cycle, acting as key regulators of many developmental processes. In this study, an AP2 homologue, NsAP2, was characterized from water lily (Nymphaea sp. cv. 'Yellow Prince') and is believed to be rather primitive in the evolution of the angiosperms. In situ RNA hybridization showed that NsAP2 transcript was present in all regions of the floral primordium, but had the highest level in the emerging floral organ primordium. After the differentiation of floral organs, NsAP2 was strongly expressed in sepals and petals, while low levels were found in stamens and carpels. The NsAP2 protein was suggested to be localized in the cell nucleus by onion transient expression experiment. Overexpression of NsAP2 in Arabidopsis led to more petal numbers, and Arabidopsis plants expressing NsAP2 exhibited higher plant height, which may be a result of down-regulated expression of GA2ox2 and GA2ox7. Our results indicated that the NsAP2 protein may function in flower organogenesis in water lily, and it is a promising gene for plant height improvement. PMID:22591856

  16. Genome-Wide Identification of the Target Genes of AP2-O, a Plasmodium AP2-Family Transcription Factor.

    PubMed

    Kaneko, Izumi; Iwanaga, Shiroh; Kato, Tomomi; Kobayashi, Issei; Yuda, Masao

    2015-05-01

    Stage-specific transcription is a fundamental biological process in the life cycle of the Plasmodium parasite. Proteins containing the AP2 DNA-binding domain are responsible for stage-specific transcriptional regulation and belong to the only known family of transcription factors in Plasmodium parasites. Comprehensive identification of their target genes will advance our understanding of the molecular basis of stage-specific transcriptional regulation and stage-specific parasite development. AP2-O is an AP2 family transcription factor that is expressed in the mosquito midgut-invading stage, called the ookinete, and is essential for normal morphogenesis of this stage. In this study, we identified the genome-wide target genes of AP2-O by chromatin immunoprecipitation-sequencing and elucidate how this AP2 family transcription factor contributes to the formation of this motile stage. The analysis revealed that AP2-O binds specifically to the upstream genomic regions of more than 500 genes, suggesting that approximately 10% of the parasite genome is directly regulated by AP2-O. These genes are involved in distinct biological processes such as morphogenesis, locomotion, midgut penetration, protection against mosquito immunity and preparation for subsequent oocyst development. This direct and global regulation by AP2-O provides a model for gene regulation in Plasmodium parasites and may explain how these parasites manage to control their complex life cycle using a small number of sequence-specific AP2 transcription factors. PMID:26018192

  17. Genome-Wide Identification of the Target Genes of AP2-O, a Plasmodium AP2-Family Transcription Factor

    PubMed Central

    Kaneko, Izumi; Iwanaga, Shiroh; Kato, Tomomi; Kobayashi, Issei; Yuda, Masao

    2015-01-01

    Stage-specific transcription is a fundamental biological process in the life cycle of the Plasmodium parasite. Proteins containing the AP2 DNA-binding domain are responsible for stage-specific transcriptional regulation and belong to the only known family of transcription factors in Plasmodium parasites. Comprehensive identification of their target genes will advance our understanding of the molecular basis of stage-specific transcriptional regulation and stage-specific parasite development. AP2-O is an AP2 family transcription factor that is expressed in the mosquito midgut-invading stage, called the ookinete, and is essential for normal morphogenesis of this stage. In this study, we identified the genome-wide target genes of AP2-O by chromatin immunoprecipitation-sequencing and elucidate how this AP2 family transcription factor contributes to the formation of this motile stage. The analysis revealed that AP2-O binds specifically to the upstream genomic regions of more than 500 genes, suggesting that approximately 10% of the parasite genome is directly regulated by AP2-O. These genes are involved in distinct biological processes such as morphogenesis, locomotion, midgut penetration, protection against mosquito immunity and preparation for subsequent oocyst development. This direct and global regulation by AP2-O provides a model for gene regulation in Plasmodium parasites and may explain how these parasites manage to control their complex life cycle using a small number of sequence-specific AP2 transcription factors. PMID:26018192

  18. Metallized solid rocket propellants based on AN/AP and PSAN/AP for access to space

    NASA Astrophysics Data System (ADS)

    Levi, S.; Signoriello, D.; Gabardi, A.; Molinari, M.; Galfetti, L.; Deluca, L. T.; Cianfanelli, S.; Klyakin, G. F.

    2009-09-01

    Solid rocket propellants based on dual mixes of inorganic crystalline oxidizers (ammonium nitrate (AN) and ammonium perchlorate (AP)) with binder and a mixture of micrometric-nanometric aluminum were investigated. Ammonium nitrate is a low-cost oxidizer, producing environment friendly combustion products but with lower specific impulse compared to AP. The better performance obtained with AP and the low quantity of toxic emissions obtained by using AN have suggested an interesting compromise based on a dual mixture of the two oxidizers. To improve the thermal response of raw AN, different types of phase stabilized AN (PSAN) and AN/AP co-crystals were investigated.

  19. Activator protein-2gamma (AP-2gamma) expression is specifically induced by oestrogens through binding of the oestrogen receptor to a canonical element within the 5'-untranslated region.

    PubMed Central

    Orso, Francesca; Cottone, Erika; Hasleton, Mark D; Ibbitt, J Claire; Sismondi, Piero; Hurst, Helen C; De Bortoli, Michele

    2004-01-01

    The activator protein 2 (AP-2) transcription factors are essential proteins for oestrogenic repression of the ERBB2 proto-oncogene in breast cancer cells. In the present study, we have examined the possible oestrogenic regulation of AP-2 genes themselves in breast-tumour-derived lines. As early as 1 h after oestrogen treatment, AP-2gamma mRNA was markedly increased, whereas AP-2alpha was down-regulated, but with slower kinetics, and AP-2beta was not affected at all. Addition of anti-oestrogens ablated these effects. Modulation of the protein levels corresponded to changes in the transcript levels, thus suggesting that in oestrogen-treated cells, an inversion of the balance between AP-2alpha and AP-2gamma isoforms occurs. The 5'-untranslated region (5'-UTR) of the human AP-2gamma gene contains one consensus and one degenerate oestrogen-responsive element (ERE). Reporter constructs carrying the AP-2gamma promoter and the 5'-UTR were up-regulated by oestrogens in transient transfection assays. Deletion of the most conserved (but not of the degenerate) ERE from reporter constructs abrogated the oestrogenic response, although both ERE-containing segments were footprinted in DNaseI protection assays. In vitro binding assays demonstrated the ability of oestrogen receptor alpha (ERalpha) to bind to this site, and chromatin immunoprecipitation analysis of the endogenous gene showed that ERalpha occupies this region in response to oestrogens. We conclude that AP-2gamma is a primary oestrogen-responsive gene and suggest that AP-2 proteins may mediate some oestrogenic responses. PMID:14565844

  20. Using selenomethionyl derivatives to assign sequence in low-resolution structures of the AP2 clathrin adaptor

    PubMed Central

    Kelly, Bernard T.; Graham, Stephen C.; Owen, David J.

    2016-01-01

    Selenomethionine incorporation is a powerful technique for assigning sequence to regions of electron density at low resolution. Genetic introduction of methionine point mutations and the subsequent preparation and crystallization of selenomethionyl derivatives permits unambiguous sequence assignment by enabling the placement of the anomalous scatterers (Se atoms) thus introduced. Here, the use of this approach in the assignment of sequence in a part of the AP2 clathrin adaptor complex that is responsible for clathrin binding is described. AP2 plays a pivotal role in clathrin-mediated endocytosis, a tightly regulated process in which cell-surface transmembrane proteins are internalized from the plasma membrane by incorporation into lipid-enclosed transport vesicles. AP2 binds cargo destined for internalization and recruits clathrin, a large trimeric protein that helps to deform the membrane to produce the transport vesicle. By selenomethionine labelling of point mutants, it was shown that the clathrin-binding site is buried within a deep cleft of the AP2 complex. A membrane-stimulated conformational change in AP2 releases the clathrin-binding site from autoinhibition, thereby linking clathrin recruitment to membrane localization. PMID:26960121

  1. Catalysts of DNA Strand Cleavage at Apurinic/Apyrimidinic Sites.

    PubMed

    Minko, Irina G; Jacobs, Aaron C; de Leon, Arnie R; Gruppi, Francesca; Donley, Nathan; Harris, Thomas M; Rizzo, Carmelo J; McCullough, Amanda K; Lloyd, R Stephen

    2016-01-01

    Apurinic/apyrimidinic (AP) sites are constantly formed in cellular DNA due to instability of the glycosidic bond, particularly at purines and various oxidized, alkylated, or otherwise damaged nucleobases. AP sites are also generated by DNA glycosylases that initiate DNA base excision repair. These lesions represent a significant block to DNA replication and are extremely mutagenic. Some DNA glycosylases possess AP lyase activities that nick the DNA strand at the deoxyribose moiety via a β- or β,δ-elimination reaction. Various amines can incise AP sites via a similar mechanism, but this non-enzymatic cleavage typically requires high reagent concentrations. Herein, we describe a new class of small molecules that function at low micromolar concentrations as both β- and β,δ-elimination catalysts at AP sites. Structure-activity relationships have established several characteristics that appear to be necessary for the formation of an iminium ion intermediate that self-catalyzes the elimination at the deoxyribose ring. PMID:27363485

  2. Catalysts of DNA Strand Cleavage at Apurinic/Apyrimidinic Sites

    PubMed Central

    Minko, Irina G.; Jacobs, Aaron C.; de Leon, Arnie R.; Gruppi, Francesca; Donley, Nathan; Harris, Thomas M.; Rizzo, Carmelo J.; McCullough, Amanda K.; Lloyd, R. Stephen

    2016-01-01

    Apurinic/apyrimidinic (AP) sites are constantly formed in cellular DNA due to instability of the glycosidic bond, particularly at purines and various oxidized, alkylated, or otherwise damaged nucleobases. AP sites are also generated by DNA glycosylases that initiate DNA base excision repair. These lesions represent a significant block to DNA replication and are extremely mutagenic. Some DNA glycosylases possess AP lyase activities that nick the DNA strand at the deoxyribose moiety via a β- or β,δ-elimination reaction. Various amines can incise AP sites via a similar mechanism, but this non-enzymatic cleavage typically requires high reagent concentrations. Herein, we describe a new class of small molecules that function at low micromolar concentrations as both β- and β,δ-elimination catalysts at AP sites. Structure-activity relationships have established several characteristics that appear to be necessary for the formation of an iminium ion intermediate that self-catalyzes the elimination at the deoxyribose ring. PMID:27363485

  3. Residual radiation studies at AP0

    SciTech Connect

    Alexander J Elwyn et al.

    2002-06-19

    The radiation environment at the NuMI experiment has received a lot of attention in the last few years in preparation for project construction. One important issue is the induced radioactivation of the components in the beam line and the shielding materials. This arises from irradiation by hadrons that are generated in the target. Since the level of the residual activity has to be considered when determining access procedures for maintenance during NuMI operation, an understanding of the properties of the remnant radiation is important. To this end, experimental studies were performed in the target vault at AP0 which is similar in design to the NuMI target area. Here 120 GeV protons bombard a target, generating the hadrons that produce the induced radioactivity. Two sets of samples each consisting of three small cylindrical or rectangular solids of iron and steel, one sample of aluminum, and one of concrete were irradiated. One set was hung just below the bottom of a module near the lithium lens (in-vault), and the other was placed on top of the modules downstream of this location (above-vault), just beneath the movable concrete roof of the vault at AP0. Further, four thin activation foils of Au, Au+Cd, In, and Al (along with small disks of the same iron, aluminum, and concrete samples discussed above) were mounted on four 10.2 cm diameter Al disks, one placed on the in-vault module and three at above-vault downstream locations as well. The radioactivity of all these materials on the 10.2 cm Al disks was determined at the Radioisotope Analysis Facility in an attempt to characterize the radionuclides produced during irradiation. The activities of the thin foils were employed in an effort to unfold a spectrum of the neutrons produced during the hadronic cascades in the target. The MARS Monte Carlo code (MO95, MO00) was used to predict and analyze the residual radiation produced during the beam irradiation. New subroutines have been developed for the MARS14 version

  4. AP-102/104 Retrieval control system qualification test procedure

    SciTech Connect

    RIECK, C.A.

    1999-05-18

    This Qualification Test Procedure documents the results of the qualification testing that was performed on the Project W-211, ''Initial Tank Retrieval Systems,'' retrieval control system (RCS) for tanks 241-AP-102 and 241-AP-104. The results confirm that the RCS has been programmed correctly and that the two related hardware enclosures have been assembled in accordance with the design documents.

  5. Is the Shine off the A.P. Apple?

    ERIC Educational Resources Information Center

    Hurwitz, Nina; Hurwitz, Sol

    2003-01-01

    Describes challenges facing College Board's efforts to expand Advanced Placement (A.P.) courses to provide equal access to previously underserved low-performing urban and rural school students while maintaining the program's high academic standards. Includes list of strategies school boards can use to achieve greater access to A.P. courses while…

  6. AP233: An Information Model for Systems Engineering

    NASA Technical Reports Server (NTRS)

    Siebes, Georg

    2009-01-01

    In today's world, information is abundant. We have no problems generating it. But we are challenged to find, organize, and exchange information. center dot A standardized model of information can help. Such a model nearly completed its development for Systems Engineering. It is referred to as AP233 (AP = Application Protocol).

  7. Status of the Advanced Photon Source (APS) linear accelerator

    SciTech Connect

    White, M.; Berg, W.; Fuja, R.; Grelick, A.; Mavrogenes, G.; Nassiri, A.; Russell, T.; Wesolowski, W.

    1993-08-01

    A 2856-MHz S-band, 450-MeV electron/positron linear accelerator is the first part of the injector for the Advanced Photon Source (APS) 7-GeV storage ring. Construction of the APS linac is currently nearing completion, and commissioning will begin in July 1993. The linac and its current status are discussed in this paper.

  8. Raising Rigor, Getting Results: Lessons Learned from AP Expansion

    ERIC Educational Resources Information Center

    Wakelyn, David

    2009-01-01

    Advanced Placement (AP), which enables high school students to take introductory college-level courses, is the nation's oldest example of a rigorous, common curriculum. Students who score well on AP exams are more likely to persist in college and earn a degree. The Advanced Placement Expansion project of the National Governors Association Center…

  9. The APS SASE FEL : modeling and code comparison.

    SciTech Connect

    Biedron, S. G.

    1999-04-20

    A self-amplified spontaneous emission (SASE) free-electron laser (FEL) is under construction at the Advanced Photon Source (APS). Five FEL simulation codes were used in the design phase: GENESIS, GINGER, MEDUSA, RON, and TDA3D. Initial comparisons between each of these independent formulations show good agreement for the parameters of the APS SASE FEL.

  10. AP-42 ADDITIONS AND REVISIONS - RUBBER PRODUCTS MANUFACTURING

    EPA Science Inventory

    This project develops emission factors, etc., for the rubber products industry which are appended to AP-42. AP42 is a massive collection of material which describes processes which generate air emissions and presents emission factors and control effectiveness information. As res...

  11. A Closer Examination of the Academic Benefits of AP

    ERIC Educational Resources Information Center

    McKillip, Mary E. M.; Rawls, Anita

    2013-01-01

    The authors sought to better understand the relationship between students participating in the Advanced Placement (AP) program and subsequent performance on the Scholastic Aptitude Test (SAT). Focusing on students graduating from U.S. public high schools in 2010, the authors used propensity scores to match junior year AP examinees in 3 subjects to…

  12. AP-42 ADDITIONS AND REVISIONS - LANDFILLS (COMBUSTION CONTROLS)

    EPA Science Inventory

    This project develops emission factors, etc., for landfills, in particular for combustion devices fed by landfill gas, for incorporation into AP-42. AP-42 is a massive collection of information concerning processes which generate air emissions and presents emission factors and co...

  13. Performance of Project Advance Students on the AP Biology Examination.

    ERIC Educational Resources Information Center

    Mercurio, Joseph; And Others

    1984-01-01

    Compared performance of Project Advance biology students (N=60) with Advanced Placement (AP) candidates (N=15,947) nationally on College Entrance Examination Board AP biology test. The research, conducted to determine comparability of the program as valid measures of academic achievement, determined that Project Advance students scored above the…

  14. Data-Based Decision Making: The Road to AP Equity

    ERIC Educational Resources Information Center

    Edwards, Kelcey; Duggan, Odette

    2012-01-01

    Presented at the Advanced Placement Annual Conference (APAC) in Lake Buena Vista, FL in July 2012. This presentation reviews concepts central to achieving equitable AP access and success for all willing and academically prepared students. We analyze trends in participation and performance by race/ethnicity from the AP Report to the Nation and…

  15. GPI-AP release in cellular, developmental, and reproductive biology.

    PubMed

    Fujihara, Yoshitaka; Ikawa, Masahito

    2016-04-01

    Glycosylphosphatidylinositol-anchored proteins (GPI-APs) contain a covalently linked GPI anchor located on outer cell membranes. GPI-APs are ubiquitously conserved from protozoa to vertebrates and are critical for physiological events such as development, immunity, and neurogenesis in vertebrates. Both membrane-anchored and soluble GPI-APs play a role in regulating their protein conformation and functional properties. Several pathways mediate the release of GPI-APs from the plasma membrane by vesiculation or cleavage. Phospholipases and putative substrate-specific GPI-AP-releasing enzymes, such as NOTUM, glycerophosphodiesterase 2, and angiotensin-converting enzyme, have been characterized in mammals. Here, the protein modifications resulting from the cleavage of the GPI anchor are discussed in the context of its physiological functions. PMID:26593072

  16. The AP Exam: Is This How People Learn?

    NASA Astrophysics Data System (ADS)

    Eisenkraft, Arthur

    2001-04-01

    The AP Physics course is often the most rigorous experience a student will have in high school. The culminating AP exam tests problem solving skills using traditional questions that are found in most freshmen books. The nature of the test imparts a specific focus on the course, which in turn shapes the students' impression of physics. Over the past decade, we have been discovering a great deal about how people learn in general. We have also learned specifically about the inadequacies of traditional physics instruction. Has the AP exam kept pace with this new knowledge? Can the AP exam be altered in order to require a different type of instruction that is informed by cognitive research? What would the new exam look like? Samples of exam questions from AP Exams will be viewed through the lens of cognitive research. Alternative physics challenges will be presented and the difficulties in grading these questions noted.

  17. Evolutionary period changes in the rapidly oscillating Ap stars

    NASA Technical Reports Server (NTRS)

    Heller, Clayton H.; Kawaler, Steven D.

    1988-01-01

    Preliminary computations of the asymptotic p-mode frequency spacings, and their time derivatives, are presented for a grid of evolutionary models of the rapidly oscillating Ap ('roAp') stars. It is shown how the frequency spacings depend on stellar mass, metallicity, and evolutionary stage. The frequency spacings observed in two of the roAp stars indicate that they either are main-sequence objects or have evolved off of the main sequence, depending on how the pulsation spectrum is interpreted. A way to remove this ambiguity is suggested: secular evolutionary changes in the pulsation frequencies may be measurable if the roAp stars have evolved off of the main sequence. In principle, this determination may be made for the roAp star HR 1217 with existing data.

  18. Top-up operation experience at APS.

    SciTech Connect

    Emery, L.

    1999-03-31

    The Advanced Photon Source (APS) is a 7-OeV, third-generation synchrotrons radiation source. To provide more stable beam for users, in September 1998 we began commissioning a new operating mode called ''top-up.'' In this mode, the beam current does not decay but is maintained at a high level using frequent injection, while photon shutters are open and photon beams are delivered to users. The hardware, software, and safety requirements for top-up will be reported. Safety issues related to injection with open photon shutters are covered in companion papers in this conference. Recent operational experience includes testing aspects of top-up injection and delivering beam to X-ray users for a few hours with fractional current stability of 10{sup {minus}3}. We expect to run several top-up operation shifts in Spring 1999. Issues of importance are orbit and emittance transients during the injection and scheduling of injection pulses for the convenience of users.

  19. High Pressure Reverse Flow APS Engine

    NASA Technical Reports Server (NTRS)

    Senneff, J. M.

    1972-01-01

    A design and test demonstration effort was undertaken to evaluate the concept of the reverse flow engine for the APS engine application. The 1500 lb (6672 N) thrust engine was designed to operate on gaseous hydrogen and gaseous oxygen propellants at a mixture ratio of 4 and to achieve the objective performance of 435 sec (4266 Nsec/kg) specific impulse. Superimposed durability requirements called for a million-cycle capability with 50 hours duration. The program was undertaken as a series of tasks including the initial preliminary design, design of critical test components and finally, the design and demonstration of an altitude engine which could be used interchangeably to examine operating parameters as well as to demonstrate the capability of the concept. The program results are reported with data to indicate that all of the program objectives were met or exceeded within the course of testing on the program. The analysis effort undertaken is also reported in detail and supplemented with test data in some cases where prior definitions could not be made. The results are contained of these analyses as well as the test results conducted throughout the course of the program. Finally, the test data and analytical results were combined to allow recommendations for a flight weight design. This preliminary design effort is also detailed.

  20. Structure of the endonuclease IV homologue from Thermotoga maritima in the presence of active-site divalent metal ions

    SciTech Connect

    Tomanicek, Stephen J.; Hughes, Ronny C.; Ng, Joseph D.; Coates, Leighton

    2010-10-05

    The most frequent lesion in DNA is at apurinic/apyrimidinic (AP) sites resulting from DNA-base losses. These AP-site lesions can stall DNA replication and lead to genome instability if left unrepaired. The AP endonucleases are an important class of enzymes that are involved in the repair of AP-site intermediates during damage-general DNA base-excision repair pathways. These enzymes hydrolytically cleave the 5{prime}-phosphodiester bond at an AP site to generate a free 3{prime}-hydroxyl group and a 5{prime}-terminal sugar phosphate using their AP nuclease activity. Specifically, Thermotoga maritima endonuclease IV is a member of the second conserved AP endonuclease family that includes Escherichia coli endonuclease IV, which is the archetype of the AP endonuclease superfamily. In order to more fully characterize the AP endonuclease family of enzymes, two X-ray crystal structures of the T. maritima endonuclease IV homologue were determined in the presence of divalent metal ions bound in the active-site region. These structures of the T. maritima endonuclease IV homologue further revealed the use of the TIM-barrel fold and the trinuclear metal binding site as important highly conserved structural elements that are involved in DNA-binding and AP-site repair processes in the AP endonuclease superfamily.

  1. Predictive Validity Study of the APS Writing and Reading Tests [and] Validating Placement Rules for the APS Writing Test.

    ERIC Educational Resources Information Center

    College of the Canyons, Valencia, CA. Office of Institutional Development.

    California's College of the Canyons has used the College Board Assessment and Placement Services (APS) test to assess students' abilities in basic and college English since spring 1993. These two reports summarize data from a May 1994 study of the predictive validity of the APS writing and reading tests and a June 1994 effort to validate the cut…

  2. Thermal Decomposition Characteristics of Orthorhombic Ammonium Perchlorate (o-AP) and an 0-AP/HTPB-Based Propellant

    SciTech Connect

    BEHRENS JR.,RICHARD; MINIER,LEANNA M.G.

    1999-10-25

    A study to characterize the low-temperature reactive processes for o-AP and an AP/HTPB-based propellant (class 1.3) is being conducted in the laboratory using the techniques of simultaneous thermogravimetric modulated beam mass spectrometry (STMBMS) and scanning electron microscopy (SEM). The results presented in this paper are a follow up of the previous work that showed the overall decomposition to be complex and controlled by both physical and chemical processes. The decomposition is characterized by the occurrence of one major event that consumes up to {approx}35% of the AP, depending upon particle size, and leaves behind a porous agglomerate of AP. The major gaseous products released during this event include H{sub 2}O, O{sub 2}, Cl{sub 2}, N{sub 2}O and HCl. The recent efforts provide further insight into the decomposition processes for o-AP. The temporal behaviors of the gas formation rates (GFRs) for the products indicate that the major decomposition event consists of three chemical channels. The first and third channels are affected by the pressure in the reaction cell and occur at the surface or in the gas phase above the surface of the AP particles. The second channel is not affected by pressure and accounts for the solid-phase reactions characteristic of o-AP. The third channel involves the interactions of the decomposition products with the surface of the AP. SEM images of partially decomposed o-AP provide insight to how the morphology changes as the decomposition progresses. A conceptual model has been developed, based upon the STMBMS and SEM results, that provides a basic description of the processes. The thermal decomposition characteristics of the propellant are evaluated from the identities of the products and the temporal behaviors of their GFRs. First, the volatile components in the propellant evolve from the propellant as it is heated. Second, the hot AP (and HClO{sub 4}) at the AP-binder interface oxidize the binder through reactions that

  3. Raising the acceptance of the AP2-line

    SciTech Connect

    Trbojevic, D.

    1989-04-05

    The 120 GeV Main Ring proton beam collides with the target at the end of the AP-1 line and creates antiprotons and other secondary particles. The AP-2 line transfers the negative particles from the target to the Debuncher. To provide a bigger antiproton stack size in the Accumulator, both the Debuncher as well as the AP-2 line acceptance have to be raised. This is a proposal for the improvement of the AP-2 line acceptance. The first part of the memo presents an acceptance examination of the existing AP-2 line by computer simulation, while the second presents a short proposal for aperture corrections. The computer program TURTLE was used to trace antiprotons through the AP-2 line without taking into account other negative charged particles. Betatron functions were obtained from the output of the SYNCH computer program. The SYNCH program was also used to check the dispersion match between the AP-2 line and the Debuncher. 3 refs., 6 figs., 5 tabs.

  4. Enhancing and Archiving the APS Catalog of the POSS I

    NASA Technical Reports Server (NTRS)

    Humphreys, Roberta M.

    2003-01-01

    We have worked on two different projects: 1) Archiving the APS Catalog of the POSS I for distribution to NASA's NED at IPAC, SIMBAD in France, and individual astronomers and 2) The automated morphological classification of galaxies. We have completed archiving the Catalog into easily readable binary files. The database together with the software to read it has been distributed on DVD's to the national and international data centers and to individual astronomers. The archived Catalog contains more than 89 million objects in 632 fields in the first epoch Palomar Observatory Sky Survey. Additional image parameters not available in the original on-line version are also included in the archived version. The archived Catalog is also available and can be queried at the APS web site (URL: http://aps.umn.edu) which has been improved with a much faster and more efficient querying system. The Catalog can be downloaded as binary datafiles with the source code for reading it. It is also being integrated into the SkyQuery system which includes the Sloan Digital Sky Survey, 2MASS, and the FIRST radio sky survey. We experimented with different classification algorithms to automate the morphological classification of galaxies. This is an especially difficult problem because there are not only a large number of attributes or parameters and measurement uncertainties, but also the added complication of human disagreement about the adopted types. To solve this problem we used 837 galaxy images from nine POSS I fields at the North Galactic Pole classified by two independent astronomers for which they agree on the morphological types. The initial goal was to separate the galaxies into the three broad classes relevant to issues of large scale structure and galaxy formation and evolution: early (ellipticals and lenticulars), spirals, and late (irregulars) with an accuracy or success rate that rivals the best astronomer classifiers. We also needed to identify a set of parameters derived

  5. TBC1D5 and the AP2 complex regulate ATG9 trafficking and initiation of autophagy

    PubMed Central

    Popovic, Doris; Dikic, Ivan

    2014-01-01

    The RabGAP protein TBC1D5 controls cellular endomembrane trafficking processes and binds the retromer subunit VPS29 and the ubiquitin-like protein ATG8 (LC3). Here, we describe that TBC1D5 also associates with ATG9 and the active ULK1 complex during autophagy. Moreover, ATG9 and TBC1D5 interact with clathrin and the AP2 complex. Depletion of TBC1D5 leads to missorting of ATG9 to late endosomes upon activation of autophagy, whereas inhibition of clathrin-mediated endocytosis or AP2 depletion alters ATG9 trafficking and its association with TBC1D5. Taken together, our data show that TBC1D5 and the AP2 complex are important novel regulators of the rerouting of ATG9-containing vesicular carriers toward sites of autophagosome formation. PMID:24603492

  6. Development of Emergency Response Guidelines (ERGs) for AP1000

    SciTech Connect

    Yuichi Hayashi; Saiu, Gianfranco; Wright, Richard F.

    2006-07-01

    The AP1000 is two-loop 1100 MWe advanced pressurized water reactor (PWR) that uses passive safety features to enhance plant safety and to provide significant and measurable improvements in plant simplification, reliability, investment protection and plant costs. The AP1000 uses proven technology, which builds on over 30 years of operating PWR experience. The AP1000 final design certification has been approved by the NRC in December, 2005. The Emergency Response Guidelines (ERGs) have been developed for the AP1000. The generic ERGs for the low pressure reference PWR plant were used as the basic documents to develop the AP1000 ERGs. The AP1000 design differences from the reference plant were reviewed and reflected in the process of developing operational steps in each ERG. The AP1000 used PRA in both design and licensing. The provisions of the AP1000 PRA were also reviewed and incorporated into the ERGs Although the AP1000 design does not require operator actions for the first 72 hours after accidents, the operator actions with both safety-related and non-safety-related equipment have an important role to mitigate the consequence of accidents. In the event of a LOCA, the AP1000 safety-related passive core cooling system (PXS) provides sources of core makeup water along with an automatic depressurization system (ADS) consisting of several sets of redundant flow paths which are sequentially opened in stages to reduce the reactor coolant system (RCS) pressure in a controlled manner. The final stage of this system, ADS-4, consists of four large valves that open off the hot legs, reducing the pressure to allow gravity injection from the in-containment refueling water storage tank (IRWST) and eventually the containment sump recirculation. The AP1000 has non-safety-related normal residual heat removal system (RNS) pumps which can be used to provide low pressure pumped injection of cask loading pit and/or IRWST water and recirculation of containment water. These pumps are

  7. A new gap separation mechanism for APS insertion devices.

    SciTech Connect

    Trakhtenberg, E. M.; Tcheskidov, V.; Den Hartog, P. K.; Deriy, B.; Erdmann, M.; Makarov, O.; Moog, E. R.

    1999-10-25

    A new gap separation mechanism for use with the standard Advanced Photon Source (APS) 3.3-cm-period undulator magnetic structures has been designed and built and the first system has been installed in the APS storage ring. The system allows a minimum magnetic gap of 10 mm for use with the APS 8-mm insertion device vacuum chambers. The mechanism is a bolted steel frame structure with a simple 4-motor mechanical drive train. The control system uses servomotors with incremental rotary encoders and virtual absolute linear encoders.

  8. AP600 design certification thermal hydraulics testing and analysis

    SciTech Connect

    Hochreiter, L.E.; Piplica, E.J.

    1995-09-01

    Westinghouse Electric Corporation, in conjunction with the Department of Energy and the Electric Power Research Institute, have been developing an advanced light water reactor design; the AP600. The AP600 is a 1940 Mwt, 600Mwe unit which is similar to a Westinghouse two-loop Pressurized Water Reactor. The accumulated knowledge on reactor design to reduce the capital costs, construction time, and the operational and maintenance cost of the unit once it begins to generate electrical power. The AP600 design goal is to maintain an overall cost advantage over fossil generated electrical power.

  9. Purification and characterization of a novel UV lesion-specific DNA glycosylase/AP lyase from Bacillus sphaericus.

    PubMed

    Vasquez, D A; Nyaga, S G; Lloyd, R S

    2000-05-31

    The purification and characterization of a pyrimidine dimer-specific glycosylase/AP lyase from Bacillus sphaericus (Bsp-pdg) are reported. Bsp-pdg is highly specific for DNA containing the cis-syn cyclobutane pyrimidine dimer, displaying no detectable activity on oligonucleotides with trans-syn I, trans-syn II, (6-4), or Dewar photoproducts. Like other glycosylase/AP lyases that sequentially cleave the N--glycosyl bond of the 5' pyrimidine of a cyclobutane pyrimidine dimer, and the phosphodiester backbone, this enzyme appears to utilize a primary amine as the attacking nucleophile. The formation of a covalent enzyme-DNA imino intermediate is evidenced by the ability to trap this protein-DNA complex by reduction with sodium borohydride. Also consistent with its AP lyase activity, Bsp-pdg was shown to incise an AP site-containing oligonucleotide, yielding beta- and delta-elimination products. N-terminal amino acid sequence analysis of this 26 kDa protein revealed little amino acid homology to any previously reported protein. This is the first report of a glycosylase/AP lyase enzyme from Bacillus sphaericus that is specific for cis-syn pyrimidine dimers. PMID:10844244

  10. Antioxidant-induced changes of the AP-1 transcription complex are paralleled by a selective suppression of human papillomavirus transcription.

    PubMed Central

    Rösl, F; Das, B C; Lengert, M; Geletneky, K; zur Hausen, H

    1997-01-01

    Considering the involvement of a redox-regulatory pathway in the expression of human papillomaviruses (HPVs), HPV type 16 (HPV-16)-immortalized human keratinocytes were treated with the antioxidant pyrrolidine-dithiocarbamate (PDTC). PDTC induces elevated binding of the transcription factor AP-1 to its cognate recognition site within the viral regulatory region. Despite of increased AP-1 binding, normally indispensable for efficient HPV-16 transcription, viral gene expression was selectively suppressed at the level of initiation of transcription. Electrophoretic mobility supershift assays showed that the composition of the AP-1 complex, predominantly consisting of Jun homodimers in untreated cells, was altered. Irrespective of enhanced c-fos expression, c-jun was phosphorylated and became primarily heterodimerized with fra-1, which was also induced after PDTC incubation. Additionally, there was also an increased complex formation between c-jun and junB. Because both fra-1 and junB overexpression negatively interferes with c-jun/c-fos trans-activation of AP-1-responsive genes, our results suggest that the observed block in viral transcription is mainly the consequence of an antioxidant-induced reconstitution of the AP-1 transcription complex. Since expression of the c-jun/c-fos gene family is tightly regulated during cellular differentiation, defined reorganization of a central viral transcription factor may represent a novel mechanism controlling the transcription of pathogenic HPVs during keratinocyte differentiation and in the progression to cervical cancer. PMID:8985358

  11. Baculovirus p35 gene is oppositely regulated by P53 and AP-1 like factors in Spodoptera frugiperda

    SciTech Connect

    Mohareer, Krishnaveni; Sahdev, Sudhir; Hasnain, Seyed E.

    2011-11-04

    Highlights: Black-Right-Pointing-Pointer Baculovirus p35 is regulated by both viral and host factors. Black-Right-Pointing-Pointer Baculovirus p35 is negatively regulated by SfP53-like factor. Black-Right-Pointing-Pointer Baculovirus p35 is positively regulated by SfAP-1-like factor. -- Abstract: Baculovirus p35 belongs to the early class of genes of AcMNPV and requires viral factors like Immediate Early protein-1 for its transcription. To investigate the role of host factors in regulating p35 gene expression, the putative transcription factor binding sites were examined in silico and the role of these factors in influencing the transcription of p35 gene was assessed. We focused our studies on AP-1 and P53-like factors, which are activated under oxidative stress conditions. The AP-1 motif is located at -1401 while P53 motif is at -1912 relative to p35 translation start site. The predicted AP-1 and P53 elements formed specific complexes with Spodoptera frugiperda nuclear extracts. Both AP-1 and P53 motif binding proteins were down regulated as a function of AcMNPV infection in Spodoptera cells. To address the question whether during an oxidative outburst, the p35 transcription is enhanced; we investigated the role of these oxidative stress induced host transcription factors in influencing p35 gene transcription. Reporter assays revealed that AP-1 element enhances the transcription of p35 by a factor of two. Interestingly, P53 element appears to repress the transcription of p35 gene.

  12. Human Immunodeficiency Virus Type 2 (HIV-2) Gag Is Trafficked in an AP-3 and AP-5 Dependent Manner

    PubMed Central

    Alford, Justine E.; Marongiu, Michela; Watkins, Gemma L.

    2016-01-01

    Although human immunodeficiency virus (HIV) types 1 and 2 are closely related lentiviruses with similar replication cycles, HIV-2 infection is associated with slower progression to AIDS, a higher proportion of long term non-progressors, and lower rates of transmission than HIV-1, likely as a consequence of a lower viral load during HIV-2 infection. A mechanistic explanation for the differential viral load remains unclear but knowledge of differences in particle production between HIV-1 and HIV-2 may help to shed light on this issue. In contrast to HIV-1, little is known about the assembly of HIV-2 particles, and the trafficking of HIV-2 Gag, the structural component of the virus, within cells. We have established that HIV-2 Gag accumulates in intracellular CD63 positive compartments, from which it may be delivered or recycled to the cell surface, or degraded. HIV-2 particle release was dependent on the adaptor protein complex AP-3 and the newly identified AP-5 complex, but much less so on AP-1. In contrast, HIV-1 particle release required AP-1 and AP-3, but not AP-5. AP-2, an essential component of clathrin-mediated endocytosis, which was previously shown to be inhibitory to HIV-1 particle release, had no effect on HIV-2. The differential requirement for adaptor protein complexes confirmed that HIV-1 and HIV-2 Gag have distinct cellular trafficking pathways, and that HIV-2 particles may be more susceptible to degradation prior to release. PMID:27392064

  13. Differential recognition of a dileucine-based sorting signal by AP-1 and AP-3 reveals a requirement for both BLOC-1 and AP-3 in delivery of OCA2 to melanosomes

    PubMed Central

    Sitaram, Anand; Dennis, Megan K.; Chaudhuri, Rittik; De Jesus-Rojas, Wilfredo; Tenza, Danièle; Setty, Subba Rao Gangi; Wood, Christopher S.; Sviderskaya, Elena V.; Bennett, Dorothy C.; Raposo, Graça; Bonifacino, Juan S.; Marks, Michael S.

    2012-01-01

    Cell types that generate unique lysosome-related organelles (LROs), such as melanosomes in melanocytes, populate nascent LROs with cargoes that are diverted from endosomes. Cargo sorting toward melanosomes correlates with binding via cytoplasmically exposed sorting signals to either heterotetrameric adaptor AP-1 or AP-3. Some cargoes bind both adaptors, but the relative contribution of each adaptor to cargo recognition and their functional interactions with other effectors during transport to melanosomes are not clear. Here we exploit targeted mutagenesis of the acidic dileucine–based sorting signal in the pigment cell–specific protein OCA2 to dissect the relative roles of AP-1 and AP-3 in transport to melanosomes. We show that binding to AP-1 or AP-3 depends on the primary sequence of the signal and not its position within the cytoplasmic domain. Mutants that preferentially bound either AP-1 or AP-3 each trafficked toward melanosomes and functionally complemented OCA2 deficiency, but AP-3 binding was necessary for steady-state melanosome localization. Unlike tyrosinase, which also engages AP-3 for optimal melanosomal delivery, both AP-1– and AP-3–favoring OCA2 variants required BLOC-1 for melanosomal transport. These data provide evidence for distinct roles of AP-1 and AP-3 in OCA2 transport to melanosomes and indicate that BLOC-1 can cooperate with either adaptor during cargo sorting to LROs. PMID:22718909

  14. QCD on the Massively Parallel Computer AP1000

    NASA Astrophysics Data System (ADS)

    Akemi, K.; Fujisaki, M.; Okuda, M.; Tago, Y.; Hashimoto, T.; Hioki, S.; Miyamura, O.; Takaishi, T.; Nakamura, A.; de Forcrand, Ph.; Hege, C.; Stamatescu, I. O.

    We present the QCD-TARO program of calculations which uses the parallel computer AP1000 of Fujitsu. We discuss the results on scaling, correlation times and hadronic spectrum, some aspects of the implementation and the future prospects.

  15. Engaging Cuban Physicists Through the APS/CPS Partnership

    NASA Astrophysics Data System (ADS)

    Lerch, Irving A.; Lerch, Irving A.

    In his reflections on Cuban physics, Marcelo Alonso urges APS to take steps to promote interactions between Cuban and US physicists. As an introduction to Marcello's essay, this note will summarize past and current activities.

  16. An improved model for the combustion of AP composite propellants

    NASA Technical Reports Server (NTRS)

    Cohen, N. S.; Strand, L. D.

    1981-01-01

    This paper presents several improvements to the BDP model of steady-state burning of AP composite solid propellants. The Price-Boggs-Derr model of AP monopropellant burning is incorporated to represent the AP. A separate energy equation is written for the binder to permit a different surface temperature from the AP; this includes an analysis of the sharing of primary diffusion flame energy, and correction of a BDP model inconsistency in treating the binder regression rate. A method for assembling component contributions to calculate the burning rates of multimodal propellants is also presented. Results are shown in the form of representative burning rate curves, comparisons with data, and calculated internal details of interest. Ideas for future work are discussed in an Appendix.

  17. A hot-spare injector for the APS linac.

    SciTech Connect

    Lewellen, J. W.

    1999-04-13

    Last year a second-generation SSRL-type thermionic cathode rf gun was installed in the Advanced Photon Source (APS) linac. This gun (referred to as ''gun2'') has been successfully commissioned and now serves as the main injector for the APS linac, essentially replacing the Koontz-type DC gun. To help ensure injector availability, particularly with the advent of top-up mode operation at the APS, a second thermionic-cathode rf gun will be installed in the APS linac to act as a hot-spare beam source. The hot-spare installation includes several unique design features, including a deep-orbit Panofsky-style alpha magnet. Details of the hot-spare beamline design and projected performance are presented, along with some plans for future performance upgrades.

  18. Structure of the two-domain hexameric APS kinase from Thiobacillus denitrificans: structural basis for the absence of ATP sulfurylase activity

    SciTech Connect

    Gay, Sean C.; Segel, Irwin H.; Fisher, Andrew J.

    2009-10-01

    APS kinase from Thiobacillus denitrificans contains an inactive N-terminal ATP sulfurylase domain. The structure presented unveils the first hexameric assembly for an APS kinase, and reveals that structural changes in the N-terminal domain disrupt the ATP sulfurylase active site thus prohibiting activity. The Tbd-0210 gene of the chemolithotrophic bacterium Thiobacillus denitrificans is annotated to encode a 60.5 kDa bifunctional enzyme with ATP sulfurylase and APS kinase activity. This putative bifunctional enzyme was cloned, expressed and structurally characterized. The 2.95 Å resolution X-ray crystal structure reported here revealed a hexameric assembly with D{sub 3} symmetry. Each subunit contains a large N-terminal sulfurylase-like domain and a C-terminal APS kinase domain reminiscent of the two-domain fungal ATP sulfurylases of Penicillium chrysogenum and Saccharomyces cerevisiae, which also exhibit a hexameric assembly. However, the T. denitrificans enzyme exhibits numerous structural and sequence differences in the N-terminal domain that render it inactive with respect to ATP sulfurylase activity. Surprisingly, the C-terminal domain does indeed display APS kinase activity, indicating that this gene product is a true APS kinase. Therefore, these results provide the first structural insights into a unique hexameric APS kinase that contains a nonfunctional ATP sulfurylase-like domain of unknown function.

  19. Divisions Iv-V / Working Group ap & Related Stars

    NASA Astrophysics Data System (ADS)

    Mathys, Gautier; Cunha, Margarida; Dworetsky, Michael; Kochukhov, Oleg; Kupka, Friedrich; LeBlanc, Francis; Monier, Richard; Paunzen, Ernst; Pintado, Olga; Piskunov, Nikolai; Ziznovsky, Jozef

    2012-04-01

    The purpose of the Working Group on Ap and Related Stars (ApWG) is to promote and facilitate research about stars in the spectral type range from B to early F that exhibit surface chemical peculiarities and related phenomena. This is a very active field of research, in which a wide variety of new developments have taken place since 2009, as illustrated by the following selected highlights.

  20. Rat beta 1-adrenergic receptor regulatory region containing consensus AP-2 elements recognizes novel transactivator proteins.

    PubMed

    Kirigiti, P; Yang, Y F; Li, X; Li, B; Midson, C N; Machida, C A

    2000-03-01

    beta 1-Adrenergic receptors (beta1-ARs) serve as important regulators of central nervous system (CNS)-mediated behavior and several neural functions, including mood, memory, neuroendocrine control, and stimulation of autonomic function. Using beta 1-AR-luciferase reporter recombinants, we have previously determined that important beta 1-AR genetic elements controlling expression within the C6 glioma cell line are contained within the region -396 to -299, relative to the translational start site. By conducting progressive internal deletions of the rat beta 1-AR 5' flanking region and with the use of beta 1-AR-luciferase recombinants, we have verified that this region contains the primary beta 1-AR promoter and/or major regulatory elements. To begin the identification of protein factors involved in beta 1-AR transcriptional activity conferred by this beta 1-AR region and flanking sequences, we conducted electrophoretic mobility shift assays using defined beta 1-AR DNA subregion probes. One probe (GS-1), encompassing the region -396 to -367, was found to produce two major and two minor mobility shift complexes when bound to nuclear extracts from the beta 1-AR expresser C6 cell line. UV-crosslinking of DNA-protein complexes, coupled with DNase I digestion, indicated that this beta 1-AR region interacts with one major protein of approximately 117 kDa molecular weight and additional minor proteins. GS-1 DNA-protein complexes were observed using beta 1-AR expresser tissues in the CNS, including cortex, hippocampus, and olfactory bulb. No DNA-protein complexes were observed when using nuclear extracts from beta 1-AR nonexpresser tissues; in some cases, using L6 cells, previously characterized to express little or no beta1-ARs, a reduction in intensities of the DNA-protein complexes was observed. Competition experiments indicate that nuclear protein binds to one of two subregions within the GS-1 sequence that contain AP-2-like consensus elements. Recombinant AP-2 protein

  1. The AP1000{sup R} China projects move forward to construction completion and equipment installation

    SciTech Connect

    Harrop, G.

    2012-07-01

    The AP1000 design is the only Generation III+ technology to receive design certification from the U.S. Nuclear Regulatory Commission. This evolutionary design provides the highest safety and performance standards and has several distinct advantages over other designs, including improved operations and reduced construction schedule risks through the use of modern, modular, engineering principles that allow construction and fabrication tasks traditionally performed in sequence to be undertaken in parallel. Since the first granting of Design Certification in 2005 by the NRC, the AP1000 design has been modified to meet emergent NRC requirements such as those requiring the design to withstand the impact of an aircraft crash. Both domestic and foreign utilities have turned to the Westinghouse AP1000 plant design to meet their near - and long-term sustainable energy needs. The first ever deployment of this advanced U.S. nuclear power technology began in China in 2007 with the award of a contract to build four AP1000 units, constructed in pairs at the coastal sites of Sanmen (Zhejiang Province) and Haiyang (Shandong Province). Currently, all four units are at an advanced stage of construction. The commercial operation date for Sanmen Unit 1 is November 2013 followed by Haiyang Unit 1 being operational in May 2014. Construction and equipment manufacture is at an advanced stage. Sanmen Unit 1 equipment that has been delivered includes the reactor vessel, the reactor vessel closure head, the passive residual heat removal heat exchanger, the integrated head package, the polar crane, and the refueling machine. The steam generators are also completed. The RV was installed within the containment vessel building in September 2011. The installation of this major equipment will allow the setting of the containment vessel top head. Haiyang Unit 1 is also achieving significant progress. Significant benefits continue to be realized as a result of lessons learned and experience gained

  2. The (Very) Slow Rotation of Magnetic Ap Stars

    NASA Astrophysics Data System (ADS)

    Mathys, Gautier

    2015-08-01

    To this date, 34 magnetic Ap stars that have periods of variation longer than 30 days are known. They represent a considerable fraction of the total number of Ap stars whose period has been reliably determined. All the available evidence unambiguously indicates that the observed variations of those long-period Ap stars result from the changing aspect of their visible hemisphere as they rotate, thus that the oblique rotator model is applicable throughout the whole range of periods of variation of the Ap stars. We show that the periods of the most slowly rotating Ap stars must be of the order of 300 years, and that some may even be longer, possibly up to 1000 years. The 5 to 6 orders of magnitude spanned by the rotation periods of the Ap stars present a major challenge for the understanding of their origin and their evolution. To guide the theo- retical developments, observational hints may be found in possible differences between the magnetic properties of stars that have rotation periods in different ranges. Such differences are starting to emerge from the existing data. To increase their significance level, study of the longest-period stars must be continued over their full rotation cycle. Failure to secure observations now may leave critical data missing for several decades, or even centuries.

  3. Thermal Decomposition Characteristics of Orthorhombic Ammonium Perchlorate (o-AP)

    SciTech Connect

    Behrens, R.; Minier, L.

    1999-03-01

    Preliminary STMBMS and SEM results of the thermal decomposition of AP in the orthorhombic phase are presented. The overall decomposition is shown to be complex and controlled by both physical and chemical processes. The data show that the physical and chemical processes can be probed and characterized utilizing SEM and STMBMS. The overall decomposition is characterized by three distinguishing features: an induction period, and accelerator period and a deceleratory period. The major decomposition event occurs in the subsurface of the AP particles and propagates towards the center of the particle with time. The amount of total decomposition is dependent upon particle size and increases from 23% for {approximately}50{micro}m-diameter AP to 33% for {approximately}200{micro}m-diameter AP. A conceptual model of the physical processes is presented. Insight into the chemical processes is provided by the gas formation rates that are measured for the gaseous products. To our knowledge, this is the first presentation of data showing that the chemical and physical decomposition processes can be identified from one another, probed and characterized at the level that is required to better understand the thermal decomposition behavior of AP. Future work is planned with the goal of obtaining data that can be used to develop a mathematical description for the thermal decomposition of o-AP.

  4. DisAp-dependent striated fiber elongation is required to organize ciliary arrays

    PubMed Central

    Galati, Domenico F.; Bonney, Stephanie; Kronenberg, Zev; Clarissa, Christina; Yandell, Mark; Elde, Nels C.; Jerka-Dziadosz, Maria; Giddings, Thomas H.; Frankel, Joseph

    2014-01-01

    Cilia-organizing basal bodies (BBs) are microtubule scaffolds that are visibly asymmetrical because they have attached auxiliary structures, such as striated fibers. In multiciliated cells, BB orientation aligns to ensure coherent ciliary beating, but the mechanisms that maintain BB orientation are unclear. For the first time in Tetrahymena thermophila, we use comparative whole-genome sequencing to identify the mutation in the BB disorientation mutant disA-1. disA-1 abolishes the localization of the novel protein DisAp to T. thermophila striated fibers (kinetodesmal fibers; KFs), which is consistent with DisAp’s similarity to the striated fiber protein SF-assemblin. We demonstrate that DisAp is required for KFs to elongate and to resist BB disorientation in response to ciliary forces. Newly formed BBs move along KFs as they approach their cortical attachment sites. However, because they contain short KFs that are rotated, BBs in disA-1 cells display aberrant spacing and disorientation. Therefore, DisAp is a novel KF component that is essential for force-dependent KF elongation and BB orientation in multiciliary arrays. PMID:25533842

  5. A cluster region of AP-1 responsive elements is required for transcriptional activity of mouse ODC gene by hepatocyte growth factor.

    PubMed

    Bianchi, Laura; Tacchini, Lorenza; Matteucci, Emanuela; Desiderio, Maria Alfonsina

    2002-05-01

    Ornithine decarboxylase (ODC) activity is regulated by a variety of mechanisms including transcription, translation, and RNA and protein half-life. Since in mouse B16-F1 melanoma cells an early and remarkable (about 6-fold) increase in steady state mRNA levels was observed after hepatocyte growth factor (HGF) treatment, we investigated the transcriptional regulation of mouse ODC promoter. Transient transfection of various ODC-luciferase promoter constructs into the B16-Fl cells in combination with electrophoretic mobility shift assays identified the HGF-responsive element as a cluster of three AP-1 binding sites (-1660 to -1572). Even if each site differs from the canonical TPA responsive element for one nucleotide, only the first two AP-1 consensus sequences seemed to be functional since allowed DNA-binding activity of nuclear proteins after HGF treatment. Comparison of the results of transfection assays with the pOD2.5-luc (2.5 kb gene fragment) and with the construct deprived of the AP-1 cluster pOD-B-luc showed that this 50 bp region was required for ODC transactivating activity in response to HGF. Since in B16-F1 cells HGF increased AP-1 activity and the mRNA expression of various AP-1 subunits, we may conclude that HGF-induced transcription of mouse ODC was largely due to triggering of AP-1 pathway. PMID:12054494

  6. Neil DNA glycosylases promote substrate turnover by Tdg during DNA demethylation

    PubMed Central

    Arab, Khelifa; Kienhöfer, Sabine; von Seggern, Annika; Niehrs, Christof

    2016-01-01

    DNA 5-methylcytosine is a dynamic epigenetic mark which plays important roles in development and disease. In the Tet-Tdg demethylation pathway, methylated cytosine is iteratively oxidized by Tet dioxygenases and unmodified cytosine is restored via thymine DNA glycosylase (Tdg). Here we show that human NEIL1 and NEIL2 DNA glycosylases coordinate abasic site processing during TET–TDG DNA demethylation. NEIL1 and NEIL2 cooperate with TDG during base excision: TDG occupies the abasic site and is displaced by NEILs, which further process the baseless sugar, thereby stimulating TDG substrate turnover. In early Xenopus embryos Neil2 cooperates with Tdg to remove oxidized methylcytosines and to specify neural crest development together with Tet3. Thus, Neils function as AP lyases in the coordinated AP site hand-over during oxidative DNA demethylation. PMID:26751644

  7. Altered splicing of ATP6AP2 causes X-linked parkinsonism with spasticity (XPDS)

    PubMed Central

    Korvatska, Olena; Strand, Nicholas S.; Berndt, Jason D.; Strovas, Tim; Chen, Dong-Hui; Leverenz, James B.; Kiianitsa, Konstantin; Mata, Ignacio F.; Karakoc, Emre; Greenup, J. Lynne; Bonkowski, Emily; Chuang, Joseph; Moon, Randall T.; Eichler, Evan E.; Nickerson, Deborah A.; Zabetian, Cyrus P.; Kraemer, Brian C.; Bird, Thomas D.; Raskind, Wendy H.

    2013-01-01

    We report a novel gene for a parkinsonian disorder. X-linked parkinsonism with spasticity (XPDS) presents either as typical adult onset Parkinson's disease or earlier onset spasticity followed by parkinsonism. We previously mapped the XPDS gene to a 28 Mb region on Xp11.2–X13.3. Exome sequencing of one affected individual identified five rare variants in this region, of which none was missense, nonsense or frame shift. Using patient-derived cells, we tested the effect of these variants on expression/splicing of the relevant genes. A synonymous variant in ATP6AP2, c.345C>T (p.S115S), markedly increased exon 4 skipping, resulting in the overexpression of a minor splice isoform that produces a protein with internal deletion of 32 amino acids in up to 50% of the total pool, with concomitant reduction of isoforms containing exon 4. ATP6AP2 is an essential accessory component of the vacuolar ATPase required for lysosomal degradative functions and autophagy, a pathway frequently affected in Parkinson's disease. Reduction of the full-size ATP6AP2 transcript in XPDS cells and decreased level of ATP6AP2 protein in XPDS brain may compromise V-ATPase function, as seen with siRNA knockdown in HEK293 cells, and may ultimately be responsible for the pathology. Another synonymous mutation in the same exon, c.321C>T (p.D107D), has a similar molecular defect of exon inclusion and causes X-linked mental retardation Hedera type (MRXSH). Mutations in XPDS and MRXSH alter binding sites for different splicing factors, which may explain the marked differences in age of onset and manifestations. PMID:23595882

  8. Altered splicing of ATP6AP2 causes X-linked parkinsonism with spasticity (XPDS).

    PubMed

    Korvatska, Olena; Strand, Nicholas S; Berndt, Jason D; Strovas, Tim; Chen, Dong-Hui; Leverenz, James B; Kiianitsa, Konstantin; Mata, Ignacio F; Karakoc, Emre; Greenup, J Lynne; Bonkowski, Emily; Chuang, Joseph; Moon, Randall T; Eichler, Evan E; Nickerson, Deborah A; Zabetian, Cyrus P; Kraemer, Brian C; Bird, Thomas D; Raskind, Wendy H

    2013-08-15

    We report a novel gene for a parkinsonian disorder. X-linked parkinsonism with spasticity (XPDS) presents either as typical adult onset Parkinson's disease or earlier onset spasticity followed by parkinsonism. We previously mapped the XPDS gene to a 28 Mb region on Xp11.2-X13.3. Exome sequencing of one affected individual identified five rare variants in this region, of which none was missense, nonsense or frame shift. Using patient-derived cells, we tested the effect of these variants on expression/splicing of the relevant genes. A synonymous variant in ATP6AP2, c.345C>T (p.S115S), markedly increased exon 4 skipping, resulting in the overexpression of a minor splice isoform that produces a protein with internal deletion of 32 amino acids in up to 50% of the total pool, with concomitant reduction of isoforms containing exon 4. ATP6AP2 is an essential accessory component of the vacuolar ATPase required for lysosomal degradative functions and autophagy, a pathway frequently affected in Parkinson's disease. Reduction of the full-size ATP6AP2 transcript in XPDS cells and decreased level of ATP6AP2 protein in XPDS brain may compromise V-ATPase function, as seen with siRNA knockdown in HEK293 cells, and may ultimately be responsible for the pathology. Another synonymous mutation in the same exon, c.321C>T (p.D107D), has a similar molecular defect of exon inclusion and causes X-linked mental retardation Hedera type (MRXSH). Mutations in XPDS and MRXSH alter binding sites for different splicing factors, which may explain the marked differences in age of onset and manifestations. PMID:23595882

  9. Probing heterobivalent binding to the endocytic AP-2 adaptor complex by DNA-based spatial screening.

    PubMed

    Diezmann, F; von Kleist, L; Haucke, V; Seitz, O

    2015-08-01

    The double helical DNA scaffold offers a unique set of properties, which are particularly useful for studies of multivalency in biomolecular interactions: (i) multivalent ligand displays can be formed upon nucleic acid hybridization in a self-assembly process, which facilitates spatial screening (ii) valency and spatial arrangement of the ligand display can be precisely controlled and (iii) the flexibility of the ligand display can be adjusted by integrating nick sites and unpaired template regions. Herein we describe the use of DNA-based spatial screening for the characterization of the adaptor complex 2 (AP-2), a central interaction hub within the endocytic protein network in clathrin-mediated endocytosis. AP-2 is comprised of a core domain and two, so-called appendage domains, the α- and the β2-ear, which associate with cytoplasmatic proteins required for the formation or maturation of clathrin/AP-2 coated pits. Each appendage domain has two binding grooves which recognize distinct peptide motives with micromolar affinity. This provides opportunities for enhanced interactions with protein molecules that contain two (or more) different peptide motives. To determine whether a particular, spatial arrangement of binding motifs is required for high affinity binding we probed the distance-affinity relationships by means of DNA-programmed spatial screening with self-assembled peptide-DNA complexes. By using trimolecular and tetramolecular assemblies two different peptides were positioned in 2-22 nucleotide distance. The binding data obtained with both recombinant protein in well-defined buffer systems and native AP-2 in brain extract suggests that the two binding sites of the AP-2 α-appendage can cooperate to provide up to 40-fold enhancement of affinity compared to the monovalent interaction. The distance between the two recognized peptide motives was less important provided that the DNA duplex segments were connected by flexible, single strand segments. By

  10. Michael Acceptor Approach to the Design of New Salvinorin A-based High Affinity Ligands for the Kappa-Opioid Receptor

    PubMed Central

    Polepally, Prabhakar R.; Huben, Krzysztof; Vardy, Eyal; Setola, Vincent; Mosier, Philip D.; Roth, Bryan L.; Zjawiony, Jordan K.

    2014-01-01

    The neoclerodane diterpenoid salvinorin A is a major secondary metabolite isolated from the psychoactive plant Salvia divinorum. Salvinorin A has been shown to have high affinity and selectivity for the κ-opioid receptor (KOR). To study the ligand–receptor interactions that occur between salvinorin A and the KOR, a new series of salvinorin A derivatives bearing potentially reactive Michael acceptor functional groups at C-2 was synthesized and used to probe the salvinorin A binding site. The κ-, δ-, and μ-opioid receptor (KOR, DOR and MOR, respectively) binding affinities and KOR efficacies were measured for the new compounds. Although none showed wash-resistant irreversible binding, most of them showed high affinity for the KOR, and some exhibited dual affinity to KOR and MOR. Molecular modeling techniques based on the recently-determined crystal structure of the KOR combined with results from mutagenesis studies, competitive binding, functional assays and structure–activity relationships, and previous salvinorin A–KOR interaction models were used to identify putative interaction modes of the new compounds with the KOR and MOR. PMID:25193297

  11. Mapping of the spontaneous deletion in the Ap3d1 gene of mocha mice: fast and reliable genotyping

    PubMed Central

    Drasbek, Kim Ryun; Holm, Mai Marie; Delenclos, Marion; Jensen, Kimmo

    2008-01-01

    Background The mocha mouse carries a spontaneous deletion in the Ap3d1 gene, encoding the delta 1 subunit of the adaptor related protein complex 3, (Ap3d1), and subsequently lack the expression of functional AP-3. This leads to a deficiency in vesicle transport and storage, which affects neurotransmitter vesicle turnover and release in the central nervous system. Since the genomic sequence of the Ap3d1 gene of mocha mouse is not known, precise mapping of the deletion as well as reliable genotyping protocols are lacking. Findings We sequenced the Ap3d1 gene (HGNC GeneID: 8943) around the deletion site in the mocha mouse and revealed a 10639 bp deletion covering exon 2 to 6. Subsequently, new PCR primers were designed yielding a reliable genotyping protocol of both newborn and adult tissue. To examine the genotypes further, hippocampal neurons were cultured from mocha and control mice. Patch-clamp recordings showed that mocha neurons had a higher input resistance, and that autaptic EPSC in mocha cultures depressed faster and stronger as compared with control cultures. Conclusion Our study reports the sequence of the deleted part of the Ap3d1 gene in mocha mice, as well as a reliable PCR-based genotyping protocol. We cultured hippocampal neurons from control and mocha mice, and found a difference in input resistance of the neurons, and in the synaptic short-term plasticity of glutamatergic autapses showing a larger synaptic depression than controls. The described procedures may be useful for the future utilization of the mocha mouse as a model of defective vesicle biogenesis. Importantly, as genotyping by eye color is complicated in newborn mice, the designed protocol is so fast and reliable that newborn mice could rapidly be genotyped and hippocampal neurons dissociated and cultured, which is normally best done at P0-P2. PMID:19032734

  12. Molecular Mechanism for Age-Related Memory Loss: The Histone-Binding Protein RbAp48

    PubMed Central

    Pavlopoulos, Elias; Jones, Sidonie; Kosmidis, Stylianos; Close, Maggie; Kim, Carla; Kovalerchik, Olga; Small, Scott A.; Kandel, Eric R.

    2016-01-01

    To distinguish age-related memory loss more explicitly from Alzheimer’s disease (AD), we have explored its molecular underpinning in the dentate gyrus (DG), a subregion of the hippocampal formation thought to be targeted by aging. We carried out a gene expression study in human postmortem tissue harvested from both DG and entorhinal cortex (EC), a neighboring subregion unaffected by aging and known to be the site of onset of AD. Using expression in the EC for normalization, we identified 17 genes that manifested reliable age-related changes in the DG. The most significant change was an age-related decline in RbAp48, a histone-binding protein that modifies histone acetylation. To test whether the RbAp48 decline could be responsible for age-related memory loss, we turned to mice and found that, consistent with humans, RbAp48 was less abundant in the DG of old than in young mice. We next generated a transgenic mouse that expressed a dominant-negative inhibitor of RbAp48 in the adult forebrain. Inhibition of RbAp48 in young mice caused hippocampus-dependent memory deficits similar to those associated with aging, as measured by novel object recognition and Morris water maze tests. Functional magnetic resonance imaging studies showed that within the hippocampal formation, dysfunction was selectively observed in the DG, and this corresponded to a regionally selective decrease in histone acetylation. Up-regulation of RbAp48 in the DG of aged wild-type mice ameliorated age-related hippocampus-based memory loss and age-related abnormalities in histone acetylation. Together, these findings show that the DG is a hippocampal subregion targeted by aging, and identify molecular mechanisms of cognitive aging that could serve as valid targets for therapeutic intervention. PMID:23986399

  13. Molecular mechanism for age-related memory loss: the histone-binding protein RbAp48.

    PubMed

    Pavlopoulos, Elias; Jones, Sidonie; Kosmidis, Stylianos; Close, Maggie; Kim, Carla; Kovalerchik, Olga; Small, Scott A; Kandel, Eric R

    2013-08-28

    To distinguish age-related memory loss more explicitly from Alzheimer's disease (AD), we have explored its molecular underpinning in the dentate gyrus (DG), a subregion of the hippocampal formation thought to be targeted by aging. We carried out a gene expression study in human postmortem tissue harvested from both DG and entorhinal cortex (EC), a neighboring subregion unaffected by aging and known to be the site of onset of AD. Using expression in the EC for normalization, we identified 17 genes that manifested reliable age-related changes in the DG. The most significant change was an age-related decline in RbAp48, a histone-binding protein that modifies histone acetylation. To test whether the RbAp48 decline could be responsible for age-related memory loss, we turned to mice and found that, consistent with humans, RbAp48 was less abundant in the DG of old than in young mice. We next generated a transgenic mouse that expressed a dominant-negative inhibitor of RbAp48 in the adult forebrain. Inhibition of RbAp48 in young mice caused hippocampus-dependent memory deficits similar to those associated with aging, as measured by novel object recognition and Morris water maze tests. Functional magnetic resonance imaging studies showed that within the hippocampal formation, dysfunction was selectively observed in the DG, and this corresponded to a regionally selective decrease in histone acetylation. Up-regulation of RbAp48 in the DG of aged wild-type mice ameliorated age-related hippocampus-based memory loss and age-related abnormalities in histone acetylation. Together, these findings show that the DG is a hippocampal subregion targeted by aging, and identify molecular mechanisms of cognitive aging that could serve as valid targets for therapeutic intervention. PMID:23986399

  14. Mutagenicity, Stable DNA Adducts, and Abasic Sites Induced in Salmonella by Phananthro[3,4-b]- and Phenanthro[4,3-b]thiophenes, Sulfur Analogs of Benzo[c]phenanthrene

    EPA Science Inventory

    Sulfur-containing polycyclic aromatic hydrocarbons (thia-PAHs or thiaarenes) are common constituents of air pollution and cigarette smoke, yet little is known of the biological significance of exposure to these compounds. Some are mutagenic and carcinogenic, but only a few have ...

  15. Crystal structure of a nucleoside model for the inter-strand cross-link formed by the reaction of 2'-de-oxy-guanosine and an abasic site in duplex DNA.

    PubMed

    Catalano, Michael J; Ruddraraju, Kasi Viswanatharaju; Barnes, Charles L; Gates, Kent S

    2016-05-01

    The title compound, 9-[(2R,4S,5R)-4-hy-droxy-5-(hy-droxy-meth-yl)tetra-hydro-furan-2-yl]-2-{[(2R,4S,5R)-4-meth-oxy-5-(meth-oxy-meth-yl)tetra-hydro-furan-2-yl]amino}-1H-purin-6(9H)-one, C17H25N5O7, crystallizes with two independent mol-ecules (A and B) in the asymmetric unit. In the crystal, the guanosine moieties of mol-ecules A and B are linked by N-H⋯N and O-H⋯N hydrogen-bonding inter-actions, forming ribbons which are stacked to form columns along [100]. These columns are then linked by O-H⋯O hydrogen bonds between the ribose moieties and numerous C-H⋯O inter-actions to complete the three-dimensional structure. PMID:27308004

  16. Crystal structure of a nucleoside model for the inter­strand cross-link formed by the reaction of 2′-de­oxy­guanosine and an abasic site in duplex DNA

    PubMed Central

    Catalano, Michael J.; Ruddraraju, Kasi Viswanatharaju; Barnes, Charles L.; Gates, Kent S.

    2016-01-01

    The title compound, 9-[(2R,4S,5R)-4-hy­droxy-5-(hy­droxy­meth­yl)tetra­hydro­furan-2-yl]-2-{[(2R,4S,5R)-4-meth­oxy-5-(meth­oxy­meth­yl)tetra­hydro­furan-2-yl]amino}-1H-purin-6(9H)-one, C17H25N5O7, crystallizes with two independent mol­ecules (A and B) in the asymmetric unit. In the crystal, the guanosine moieties of mol­ecules A and B are linked by N—H⋯N and O—H⋯N hydrogen-bonding inter­actions, forming ribbons which are stacked to form columns along [100]. These columns are then linked by O—H⋯O hydrogen bonds between the ribose moieties and numerous C—H⋯O inter­actions to complete the three-dimensional structure. PMID:27308004

  17. Characterization of the human activator protein-2gamma (AP-2gamma) gene: control of expression by Sp1/Sp3 in breast tumour cells.

    PubMed Central

    Hasleton, Mark D; Ibbitt, J Claire; Hurst, Helen C

    2003-01-01

    The activator protein-2 (AP-2) family of DNA-binding transcription factors are developmentally regulated and also play a role in human neoplasia. In particular, the AP-2gamma protein has been shown to be overexpressed in a high percentage of breast tumours. In the present study, we report the complete sequence determination of the human TFAP2C gene encoding the AP-2gamma transcription factor plus the mapping of the transcription start site used in breast tumour-derived cells. The 5'-end of the gene lies within a CpG island and transcription is initiated at a single site within a classical initiator motif. We have gone on to investigate why some breast tumour-derived cell lines readily express AP-2gamma, whereas others do not, and show that the proximal promoter (+191 to -312) is differentially active in the two cell phenotypes. DNase footprinting led to the identification of three Sp1/Sp3-binding sites within this region, two of which are absolutely required both for promoter function and cell-type-specific activity. By Western blotting a panel of expressing and non-expressing breast tumour lines we show that the latter have higher levels of Sp3. Furthermore, increasing Sp3 levels in AP-2gamma-expressing cells led to the repression of AP-2gamma promoter activity, particularly when Sp3 inhibitory function was maximized through sumoylation. We propose that differences in the level and activity of Sp3 between breast tumour lines can determine the expression level of their AP-2gamma gene. PMID:12733991

  18. Structural basis for the recognition of tyrosine-based sorting signals by the μ3A subunit of the AP-3 adaptor complex.

    PubMed

    Mardones, Gonzalo A; Burgos, Patricia V; Lin, Yimo; Kloer, Daniel P; Magadán, Javier G; Hurley, James H; Bonifacino, Juan S

    2013-03-29

    Tyrosine-based signals fitting the YXXØ motif mediate sorting of transmembrane proteins to endosomes, lysosomes, the basolateral plasma membrane of polarized epithelial cells, and the somatodendritic domain of neurons through interactions with the homologous μ1, μ2, μ3, and μ4 subunits of the corresponding AP-1, AP-2, AP-3, and AP-4 complexes. Previous x-ray crystallographic analyses identified distinct binding sites for YXXØ signals on μ2 and μ4, which were located on opposite faces of the proteins. To elucidate the mode of recognition of YXXØ signals by other members of the μ family, we solved the crystal structure at 1.85 Å resolution of the C-terminal domain of the μ3 subunit of AP-3 (isoform A) in complex with a peptide encoding a YXXØ signal (SDYQRL) from the trans-Golgi network protein TGN38. The μ3A C-terminal domain consists of an immunoglobulin-like β-sandwich organized into two subdomains, A and B. The YXXØ signal binds in an extended conformation to a site on μ3A subdomain A, at a location similar to the YXXØ-binding site on μ2 but not μ4. The binding sites on μ3A and μ2 exhibit similarities and differences that account for the ability of both proteins to bind distinct sets of YXXØ signals. Biochemical analyses confirm the identification of the μ3A site and show that this protein binds YXXØ signals with 14-19 μm affinity. The surface electrostatic potential of μ3A is less basic than that of μ2, in part explaining the association of AP-3 with intracellular membranes having less acidic phosphoinositides. PMID:23404500

  19. Structural Basis for the Recognition of Tyrosine-based Sorting Signals by the μ3A Subunit of the AP-3 Adaptor Complex*

    PubMed Central

    Mardones, Gonzalo A.; Burgos, Patricia V.; Lin, Yimo; Kloer, Daniel P.; Magadán, Javier G.; Hurley, James H.; Bonifacino, Juan S.

    2013-01-01

    Tyrosine-based signals fitting the YXXØ motif mediate sorting of transmembrane proteins to endosomes, lysosomes, the basolateral plasma membrane of polarized epithelial cells, and the somatodendritic domain of neurons through interactions with the homologous μ1, μ2, μ3, and μ4 subunits of the corresponding AP-1, AP-2, AP-3, and AP-4 complexes. Previous x-ray crystallographic analyses identified distinct binding sites for YXXØ signals on μ2 and μ4, which were located on opposite faces of the proteins. To elucidate the mode of recognition of YXXØ signals by other members of the μ family, we solved the crystal structure at 1.85 Å resolution of the C-terminal domain of the μ3 subunit of AP-3 (isoform A) in complex with a peptide encoding a YXXØ signal (SDYQRL) from the trans-Golgi network protein TGN38. The μ3A C-terminal domain consists of an immunoglobulin-like β-sandwich organized into two subdomains, A and B. The YXXØ signal binds in an extended conformation to a site on μ3A subdomain A, at a location similar to the YXXØ-binding site on μ2 but not μ4. The binding sites on μ3A and μ2 exhibit similarities and differences that account for the ability of both proteins to bind distinct sets of YXXØ signals. Biochemical analyses confirm the identification of the μ3A site and show that this protein binds YXXØ signals with 14–19 μm affinity. The surface electrostatic potential of μ3A is less basic than that of μ2, in part explaining the association of AP-3 with intracellular membranes having less acidic phosphoinositides. PMID:23404500

  20. The Relationship between Advanced Placement and College Graduation. 2005 AP Study Series, Report 1

    ERIC Educational Resources Information Center

    Dougherty, Chrys; Mellor, Lynn; Jian, Shuling

    2006-01-01

    This study explores the relationship between college graduation rates and student participation and success in Advanced Placement (AP) courses and exams. We reviewed three approaches to examining this relationship: 1) comparing the college graduation rates of AP and non-AP students; 2) comparing the college graduation rate of AP and non-AP…

  1. AP Report to the Nation: A Closer Look at the Nation and Florida

    ERIC Educational Resources Information Center

    Sawtell, Ellen A.; Gillie, Jacqueline M.; Smith, Patricia Z.

    2012-01-01

    In February 2012, the College Board published The 8th Annual AP Report to the Nation. This session provides a deeper dive into key information for the United States with an emphasis on Florida, and participants hear how one school in Florida utilizes AP Potential™ to help build their AP Program. Participants also learn about AP participation and…

  2. Past, Present, and Future of AP Chemistry: A Brief History of Course and Exam Alignment Efforts

    ERIC Educational Resources Information Center

    Magrogan, Serena

    2014-01-01

    As part of the Advanced Placement (AP) Program's commitment to continually enhance alignment with current best practices in college-level learning, the AP Program is currently evaluating and redesigning courses and exams, one of which launched during the 2013-2014 academic school year: AP chemistry. The history of the AP chemistry course and…

  3. Experience with the EPICS PV Gateway at the APS.

    SciTech Connect

    Evans, K. Jr.; Smith, M.; Accelerator Systems Division

    2006-01-01

    The EPICS Process Variable Gateway has become a stable, high-performance application that provides access to process variables while minimizing the impact on critical Input-Output Controllers (IOCs) and implementing additional access security. The additional access security typically prevents write access but is highly configurable. The Advanced Photon Source (APS) currently uses 40 Gateways running on 11 workstations to provide access to the machine subnet from the offices and for the individual experimental teams. These include reverse Gateways that allow administration of all 40 APS Gateways from a single MEDM screen, even though the Gateways are running on separate networks. This administration includes starting, stopping, making and viewing reports, and viewing and editing access security files. There is one Gateway that provides process variable renaming. This paper provides an overview of the Gateways at the APS and describes the procedures that have been set up to use and administer them.

  4. A survey of Ap stars for weak longitudinal magnetic fields

    NASA Astrophysics Data System (ADS)

    Auriere, M.; Silvester, J.; Wade, G. A.; Bagnulo, S.; Donati, J. F.; Johnson, N.; Landstreet, J. D.; Lignieres, F.; Lueftinger, T.; Mouillet, M.; Paletou, F.; Petit, P.; Strasser, S.

    2004-10-01

    We are conducting a magnetic survey of a sample of about 30 spectroscopically-identified Ap stars (selected from the HD catalogue), but with faint or previously undetected magnetic fields. We use the MuSiCoS spectropolarimeter at Telescope Bernard Lyot (Pic du Midi Observatory, France) and the cross-correlation technique Least Squares Deconvolution (LSD; Donati et al. 1997). For 24 studied stars, we have obtained 21 detections of Stokes V Zeeman signatures (data quality and phase coverage may explain our lack of detection of any field in some objects). Our results suggest that all Ap stars are magnetic and, furthermore, that there may exist a minimum field strength for which Ap-type characteristics are produced.

  5. The fundamental parameters of the Ap star 78 Virginis. Could 78 Vir be a rapidly oscillating Ap star?

    NASA Astrophysics Data System (ADS)

    Perraut, K.; Cunha, M.; Brandão, I.; Loridat, J.; Mourard, D.; Meilland, A.; Nardetto, N.; McAlister, H.; ten Brummelaar, T. A.; Sturmann, J.; Sturmann, L.; Turner, N.; Farrington, C.; Vargas, N.

    2015-07-01

    Context. Determining the effective temperature of Ap stars, including the roAp stellar pulsators, is a difficult task owing to their strong magnetic field and their related spotted surfaces. It is, however, an important step towards constraining models of their complex atmosphere and testing proposed pulsation excitation mechanisms. Aims: Using the unique angular resolution provided by long-baseline visible interferometry, we aim at deriving accurate angular diameters of a number of Ap targets, so as to determine their unbiased effective temperature (Teff) and their accurate position in the Hertzsprung-Russell diagram, to estimate their mass and age, and to test non-adiabatic pulsation models. Interferometric results on four Ap stars have been published in earlier works. Here we report the results on a fifth, significantly hotter star. Methods: We observed 78 Vir with the visible spectrograph VEGA installed at the combined focus of the CHARA long-baseline optical array. We derived the limb-darkened diameter of this Ap star from our interferometric measurements. Based on photometric and spectroscopic data available in the literature, we estimated the star's bolometric flux and used it, in combination with its parallax and angular diameter, to determine the star's luminosity and effective temperature. We then used the derived fundamental parameters to perform a non-adiabatic pulsation analysis. Results: We determined a limb-darkened angular diameter of 0.346 ± 0.006 mas and deduced a linear radius of R = 2.11 ± 0.04 R⊙. Considering a bolometric flux of 2.73 ± 0.20 10-7 erg/cm2/s we obtained a luminosity of L/L⊙ = 27 ± 2 and an effective temperature of Teff = 9100 ± 190 K. The non-adiabatic pulsation modeling allows us to predict that high overtone pulsations could be excited in 78 Vir at frequencies ranging from 1.2 to 1.9 mHz, provided that the magnetic field is capable of suppressing envelope convection in the polar regions. Conclusions: Visible long

  6. Psychosocial Assessment of Artificial Pancreas (AP): Commentary and Review of Existing Measures and Their Applicability in AP Research

    PubMed Central

    Hood, Korey K.; Weissberg-Benchell, Jill; Aldred, Chris; Oliver, Nick; Laffel, Lori

    2015-01-01

    Abstract Aim: This study aimed to systematically review the evidence base for the use of existing psychological and psychosocial measures suitable for use in artificial pancreas (AP) research. Materials and Methods: This systematic review of published literature, gray literature, previous systematic reviews, and qualitative and economic studies was conducted using terms and abbreviations synonymous with diabetes, AP, and quality of life (QoL). Results: Two hundred ninety-two abstracts were identified that reported psychosocial assessment of diabetes-related technologies. Of these, nine met the inclusion criteria and were included. Only four of 103 ongoing trials evaluated psychosocial aspects as an outcome in the trial. Of these, treatment satisfaction, acceptance and use intention of AP, fear of hypoglycemia episodes, satisfaction with AP, and an unspecified QoL measure were used. Conclusions: A better understanding of the psychosocial side of AP systems and the extent to which human factors play a role in the uptake and efficient use of these systems will ultimately lead to the most benefit for people with diabetes. PMID:25549042

  7. The nuclear factor YY1 suppresses the human gamma interferon promoter through two mechanisms: inhibition of AP1 binding and activation of a silencer element.

    PubMed Central

    Ye, J; Cippitelli, M; Dorman, L; Ortaldo, J R; Young, H A

    1996-01-01

    Our group has previously reported that the nuclear factor Yin-Yang 1 (YY1), a ubiquitous DNA-binding protein, is able to interact with a silencer element (BE) in the gamma interferon (IFN-gamma) promoter region. In this study, we demonstrated that YY1 can directly inhibit the activity of the IFN-gamma promoter by interacting with multiple sites in the promoter. In cotransfection assays, a YY1 expression vector significantly inhibited IFN-gamma promoter activity. Mutation of the YY1 binding site in the native IFN-gamma promoter was associated with an increase in the IFN-gamma promoter activity. Analysis of the DNA sequences of the IFN-gamma promoter revealed a second functional YY1 binding site (BED) that overlaps with an AP1 binding site. In this element, AP1 enhancer activity was suppressed by YY1. Since the nuclear level of YY1 does not change upon cell activation, our data support a model that the nuclear factor YY1 acts to suppress basal IFN-gamma transcription by interacting with the promoter at multiple DNA binding sites. This repression can occur through two mechanisms: (i) cooperation with an as-yet-unidentified AP2-like repressor protein and (ii) competition for DNA binding with the transactivating factor AP1. PMID:8756632

  8. Synthesis and evaluation of derivatives of the proteasome deubiquitinase inhibitor b-AP15.

    PubMed

    Wang, Xin; D'Arcy, Pádraig; Caulfield, Thomas R; Paulus, Aneel; Chitta, Kasyapa; Mohanty, Chitralekha; Gullbo, Joachim; Chanan-Khan, Asher; Linder, Stig

    2015-11-01

    The ubiquitin-proteasome system (UPS) is increasingly recognized as a therapeutic target for the development of anticancer therapies. The success of the 20S proteasome core particle (20S CP) inhibitor bortezomib in the clinical management of multiple myeloma has raised the possibility of identifying other UPS components for therapeutic intervention. We previously identified the small molecule b-AP15 as an inhibitor of 19S proteasome deubiquitinase (DUB) activity. Building upon our previous data, we performed a structure-activity relationship (SAR) study on b-AP15 and identified VLX1570 as an analog with promising properties, including enhanced potency and improved solubility in aqueous solution. In silico modeling was consistent with interaction of VLX1570 with key cysteine residues located at the active sites of the proteasome DUBs USP14 and UCHL5. VLX1570 was found to inhibit proteasome deubiquitinase activity in vitro in a manner consistent with competitive inhibition. Furthermore, using active-site-directed probes, VLX1570 also inhibited proteasome DUB activity in exposed cells. Importantly, VLX1570 did not show inhibitory activity on a panel of recombinant non-proteasome DUBs, on recombinant kinases, or on caspase-3 activity, suggesting that VLX1570 is not an overtly reactive general enzyme inhibitor. Taken together, our data shows the chemical and biological properties of VLX1570 as an optimized proteasome DUB inhibitor. PMID:25854145

  9. Status of the Advanced Photon Source (APS) linear accelerator

    SciTech Connect

    White, M.; Arnold, N.; Berg, W.; Cours, A.; Fuja, R.; Grelick, A.; Ko, K.; Qian, Y.; Russell, T.; Sereno, N.

    1994-09-01

    A 2856-MHz S-band, electron-positron linear accelerator (linac) has been constructed at the Advanced Photon Source (APS). It is the source of particles and the injector for the other APS accelerators, and linac commissioning is well underway. The linac is operated 24 hours per day to support linac beam studies and rf conditioning, as well as positron accumulator ring and synchrotron commissioning studies. The design goal for accelerated positron current is 8-mA, and has been met. Maximum positron energy to date is 420-MeV, approaching the design goal of 450-MeV. The linac design and its performance are discussed.

  10. Analysis of the planning and scheduling functionality in APS systems

    NASA Astrophysics Data System (ADS)

    Steger-Jensen, Kenn; Hvolby, Hans-Henrik

    2001-10-01

    The paper discusses the basic functionality of planning and scheduling in Advanced Planning and Scheduling systems (APS). Three basic planning options - unconstrained planning, constrained planning and optimization are analyzed by use of theory and examples based on test of an APS system. Even though the planning functionality are radically improved compared to MRP and MRP II, the balance between the objectives are found to be too rigid. This conclusion is based on a number of examples, comparing the outcome of different objectives such as constraints based planning versus optimized planning.

  11. Piping benchmark problems for the Westinghouse AP600 Standardized Plant

    SciTech Connect

    Bezler, P.; DeGrassi, G.; Braverman, J.; Wang, Y.K.

    1997-01-01

    To satisfy the need for verification of the computer programs and modeling techniques that will be used to perform the final piping analyses for the Westinghouse AP600 Standardized Plant, three benchmark problems were developed. The problems are representative piping systems subjected to representative dynamic loads with solutions developed using the methods being proposed for analysis for the AP600 standard design. It will be required that the combined license licensees demonstrate that their solutions to these problems are in agreement with the benchmark problem set.

  12. Power supply control units for APS ring magnets

    SciTech Connect

    Despe, O.D.

    1990-04-15

    The APS storage ring (1104 meters) is divided into 40 sectors. Each sector has 38 magnet coils in five magnet bases. Every alternate sector has an additional quadrupole magnet for skew correction. AR the main dipole magnets, two in each sector are connected in series and fed from one power supply unit. A base is controlled by one power supply control unit (PSCU). Each PSCU is connected to the host computer via a local area network (LAN). This note discusses the hardware configuration of the typical power supply control system used by the APS magnets and the software commands supported by the PSCU.

  13. Dynamic analysis of the 7-GeV APS experiment hall foundation based on equivalent lumped parameter modeling

    SciTech Connect

    Wambsganss, M.W.

    1989-01-25

    In this technical note, mass-spring-dashpot, also referred to as equivalent lumped parameter, models are employed to model the soil-foundation interaction of two typical floor segments from the 7-GeV APS experiment hall. Equivalent lumped parameter models have the advantage of being easy to apply and of readily allowing for parameter studies. Analysis requires knowledge of certain properties of the soil including density, shear wave velocity, and Poisson's ratio, as well as knowledge of the degree of homogeneity of the underlying soil stratum. These data for the APS site were determined by a geotechnical investigation. A soil profile and pertinent data, obtained from crosshole seismic testing, are given. Natural frequencies and damping are calculated for the vertical, sliding, rocking, and coupled rocking/sliding modes of vibration. Subsequently, various corrections to account for modeling deficiencies'' are considered and their influences evaluated. The equivalent lumped parameter models were developed for machine foundations which, compared with the APS foundation, are smaller in plan dimension. Therefore, the applicability of these models in the analysis of the dynamic characteristics of the APS foundation must be established. The modeling is evaluated by applying the equivalent lumped parameter models in the analysis of large foundations for which test data exists. A comparison of theoretical and test results establishes the basis for an assessment of the applicability and accuracy of the modeling.

  14. Dynamic analysis of the 7-GeV APS experiment hall foundation based on equivalent lumped parameter modeling

    SciTech Connect

    Wambsganss, M.W.

    1989-01-25

    In this technical note, mass-spring-dashpot, also referred to as equivalent lumped parameter, models are employed to model the soil-foundation interaction of two typical floor segments from the 7-GeV APS experiment hall. Equivalent lumped parameter models have the advantage of being easy to apply and of readily allowing for parameter studies. Analysis requires knowledge of certain properties of the soil including density, shear wave velocity, and Poisson`s ratio, as well as knowledge of the degree of homogeneity of the underlying soil stratum. These data for the APS site were determined by a geotechnical investigation. A soil profile and pertinent data, obtained from crosshole seismic testing, are given. Natural frequencies and damping are calculated for the vertical, sliding, rocking, and coupled rocking/sliding modes of vibration. Subsequently, various corrections to account for modeling ``deficiencies`` are considered and their influences evaluated. The equivalent lumped parameter models were developed for machine foundations which, compared with the APS foundation, are smaller in plan dimension. Therefore, the applicability of these models in the analysis of the dynamic characteristics of the APS foundation must be established. The modeling is evaluated by applying the equivalent lumped parameter models in the analysis of large foundations for which test data exists. A comparison of theoretical and test results establishes the basis for an assessment of the applicability and accuracy of the modeling.

  15. Identification and characterization of Ref-1, a nuclear protein that facilitates AP-1 DNA-binding activity.

    PubMed Central

    Xanthoudakis, S; Curran, T

    1992-01-01

    Fos and Jun form a heterodimeric complex that regulates gene transcription by binding to the activator protein-1 (AP-1) DNA sequence motif. Previously, we demonstrated that the DNA-binding activity of Fos and Jun is regulated in vitro by a novel redox (reduction-oxidation) mechanism. Reduction of a conserved cysteine (cys) residue in the DNA-binding domains of Fos and Jun by chemical reducing agents or by a nuclear redox factor stimulates DNA-binding activity. Here, we describe purification and characterization of a 37 kDa protein (Ref-1) corresponding to the redox factor. Although Ref-1 does not bind to the AP-1 site in association with Fos and Jun, it partially copurifies with a subset of AP-1 proteins. Purified Ref-1 protein stimulates AP-1 DNA-binding activity through the conserved Cys residues in Fos and Jun, but it does not alter the DNA-binding specificity of Fos and Jun. Ref-1 may represent a novel redox component of the signal transduction processes that regulate eukaryotic gene expression. Images PMID:1537340

  16. The AP-2 complex is required for proper temporal and spatial dynamics of endocytic patches in fission yeast.

    PubMed

    de León, Nagore; Hoya, Marta; Curto, M-Angeles; Moro, Sandra; Yanguas, Francisco; Doncel, Cristina; Valdivieso, M-Henar

    2016-05-01

    In metazoans the AP-2 complex has a well-defined role in clathrin-mediated endocytosis. By contrast, its direct role in endocytosis in unicellular eukaryotes has been questioned. Here, we report co- immunoprecipitation between the fission yeast AP-2 component Apl3p and clathrin, as well as the genetic interactions between apl3Δ and clc1 and sla2Δ/end4Δ mutants. Furthermore, a double clc1 apl3Δ mutant was found to be defective in FM4-64 uptake. In an otherwise wild-type strain, apl3Δ cells exhibit altered dynamics of the endocytic sites, with a heterogeneous and extended lifetime of early and late markers at the patches. Additionally, around 50% of the endocytic patches exhibit abnormal spatial dynamics, with immobile patches and patches that bounce backwards to the cell surface, showing a pervasive effect of the absence of AP-2. These alterations in the endocytic machinery result in abnormal cell wall synthesis and morphogenesis. Our results complement those found in budding yeast and confirm that a direct role of AP-2 in endocytosis has been conserved throughout evolution. PMID:26749213

  17. Effective molarity in a nucleic acid-controlled reaction.

    PubMed

    Catalano, Michael J; Price, Nathan E; Gates, Kent S

    2016-06-01

    Positioning of reactive functional groups within a DNA duplex can enable chemical reactions that otherwise would not occur to an appreciable extent. However, few studies have quantitatively defined the extent to which the enforced proximity of reaction partners in duplex DNA can favor chemical processes. Here, we measured substantial effective molarities (as high as 25M) afforded by duplex DNA to a reaction involving interstrand cross-link formation between 2'-deoxyadenosine and a 2-deoxyribose abasic (Ap) site. PMID:27117430

  18. Functional erythroid promoters created by interaction of the transcription factor GATA-1 with CACCC and AP-1/NFE-2 elements.

    PubMed Central

    Walters, M; Martin, D I

    1992-01-01

    We have investigated interactions between the erythroid transcription factor GATA-1 and factors binding two cis-acting elements commonly linked to GATA sites in erythroid control elements. GATA-1 is present at all stages of erythroid differentiation, is necessary for erythropoiesis, and binds sites in all erythroid control elements. However, minimal promoters containing GATA-1 sites are inactive when tested in erythroid cells. Based on this observation, two erythroid cis elements, here termed CACCC and AP-1/NFE-2, were linked to GATA sites in minimal promoters. None of the elements linked only to a TATA box created an active promoter, but GATA sites linked to either CACCC or AP-1/NFE-2 elements formed strong erythroid promoters. A mutation of T to C at position -175 in the gamma-globin promoter GATA site, associated with hereditary persistence of fetal hemoglobin (HPFH), increased expression of these promoters in both fetal and adult cells. A construct bearing the beta-globin CACCC element was more active in adult and less active in fetal erythroid cells, when compared with the gamma-globin CACCC element. These studies suggest that erythroid control elements are formed by the interactions of at least three transcription factors, none of which functions alone. Images PMID:1438231

  19. Early Telegraphic News Dispatches: The Forerunner of the AP.

    ERIC Educational Resources Information Center

    Schwarzlose, Richard A.

    The origin of the Associated Press (AP) lies in the early cooperative news gathering efforts of the editors of several New York newspapers. As early as May 1846, these editors were "pooling" their energies in response to newly developed modes of communication--the wire and wireless telegraph and the trans-oceanic steamship mail services. The…

  20. AP: A Critical Examination of the Advanced Placement Program

    ERIC Educational Resources Information Center

    Sadler, Philip M., Ed.; Sonnert, Gerhard, Ed.; Tai, Robert H., Ed.; Klopfenstein, Kristin, Ed.

    2010-01-01

    With an annual yearly growth rate of 9.3 percent over the last two decades, Advanced Placement courses have become a juggernaut in American high school education. AP courses are routinely perceived as an indicator of educational rigor, and many schools push to enroll low-income or minority students in these courses in the hope of preparing them…

  1. The nature of the rapidly oscillating Ap stars' pulsations

    NASA Astrophysics Data System (ADS)

    Cunha, M. S.; Perraut, K.

    2013-12-01

    Chemically peculiar stars are stage to a wide variety of physical phenomena, including diffusion, convection, magnetism and pulsation. Progress in the understanding of these objects, through the study of their oscillations, can help us to characterize these physical phenomena and better understand the way they are coupled in stars. A number of chemically peculiar A-type stars, known as rapidly oscillating Ap (roAp) stars, have been known to exhibit high frequency oscillations since the early 80s. Despite this, the mechanism responsible for driving these oscillations is not fully understood. Currently, the most widely accepted theory states that oscillations in this class of pulsators are excited by the opacity mechanism acting on the hydrogen ionization region, in an envelope where convection has been suppressed by a strong magnetic field. Nevertheless, this theory fails to correctly predict some of the observations for this class of pulsators. In this paper we briefly review the current status of understanding of the driving of pulsations in roAp stars. In particular, we shall emphasize the comparison between predictions of nonadiabatic models of roAp stars with observations of a subset of pulsators of this class for which stringent data on global parameters are available.

  2. Intermediate-break LOCA analyses for the AP600 design

    SciTech Connect

    Boyack, B.E.; Lime, J.F.

    1995-07-01

    A postulated double-ended guillotine break of a direct-vessel-injection line in an AP600 plant has been analyzed. This event is characterized as an intermediate break loss-of-coolant accident (IBLOCA). Most of the insights regarding the response of the AP600 safety systems to the postulated accident are derived from calculations performed with the TRAC-PFl/MOD2 code. However, complementary insights derived from a scaled experiment conducted in the ROSA facility, as well as insights based upon calculations by other codes, are also presented. The key processes occurring in an AP600 during a IBLOCA are primary coolant system depressurization, inventory depletion, inventory replacement via emergency core coolant injection, continuous core cooling, and long-term decay heat rejection to the atmosphere. Based upon the calculated and experimental results, the AP600 will not experience a core heat up and will reach a safe shutdown state using only safety-class equipment. Only the early part of the long-term cooling period initiated by In-containment Refueling Water Storage Tank injection was evaluated Thus, the observation that the core is continuously cooled should be verified for the latter phase of the long-term cooling period, the interval when sump injection and containment cooling processes are important.

  3. 76 FR 82079 - AP1000 Design Certification Amendment

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-30

    ... any person. The NRC originally approved the AP1000 design certification in a final rule in 2006 (71 FR... notice of acceptance (ADAMS Accession No. ML073600743) in the Federal Register (73 FR 4926; January 28... applications: . Vogtle, Units 3 and 4......... Docket No. 05200025/6. 73 FR 33118. Bellefonte Nuclear...

  4. Time Trials--An AP Physics Challenge Lab

    ERIC Educational Resources Information Center

    Jones, David

    2009-01-01

    I have come to the conclusion that for high school physics classroom and laboratory experiences, simpler is better! In this paper I describe a very simple and effective lab experience that my AP students have thoroughly enjoyed year after year. I call this lab exercise "Time Trials." The experiment is simple in design and it is a lot of fun for…

  5. APS storage ring vacuum chamber: Section 1, Evaluation

    SciTech Connect

    Benaroya, R.; Roop, B.

    1995-07-01

    The vacuum characteristics of the APS storage ring vacuum chamber prototype, Section One (S1), is presented. The base pressure achieved was 4 {times} 10{sup {minus}11}, the welds contained no virtual or real leaks, the NeG strip mounting design and activation procedures have been determined, and S1 was found contaminated with hydrocarbons.

  6. Integrating Particulate Representations into AP Chemistry and Introductory Chemistry Courses

    ERIC Educational Resources Information Center

    Prilliman, Stephen G.

    2014-01-01

    The College Board's recently revised curriculum for advanced placement (AP) chemistry places a strong emphasis on conceptual understanding, including representations of particle phenomena. This change in emphasis is informed by years of research showing that students could perform algorithmic calculations but not explain those calculations…

  7. Automated Procurement System (APS) revised project management plan (DS-03)

    NASA Technical Reports Server (NTRS)

    Murphy, Diane R.

    1995-01-01

    The Project Plan is the governing document for the implementation of the Automated Procurement System (APS). It includes a description of the proposed system, describes the work to be done, establishes a schedule of deliverables, and discusses the major standards and procedures to be followed.

  8. 75 FR 75666 - Advanced Placement (AP) Test Fee Program

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-12-06

    ...: On September 1, 2010, we published in the Federal Register (75 FR 53681) a notice inviting... in the September 1, 2010 notice (75 FR 53682-53683). We encourage eligible applicants to submit their... Advanced Placement (AP) Test Fee Program AGENCY: Office of Elementary and Secondary Education...

  9. The New AP Chemistry Exam: Its Rationale, Content, and Scoring

    ERIC Educational Resources Information Center

    Price, Paul D.; Kugel, Roger W.

    2014-01-01

    The 2013-2014 academic year marks the rollout of the redesigned advanced placement (AP) chemistry course and exam. There have been many questions as to why the course was redesigned and how the new examination will differ from its legacy version. In this article we give a brief overview of the legacy course and examine why a redesign occurred in…

  10. APS undulator and wiggler sources: Monte-Carlo simulation

    SciTech Connect

    Xu, S.L.; Lai, B.; Viccaro, P.J.

    1992-02-01

    Standard insertion devices will be provided to each sector by the Advanced Photon Source. It is important to define the radiation characteristics of these general purpose devices. In this document,results of Monte-Carlo simulation are presented. These results, based on the SHADOW program, include the APS Undulator A (UA), Wiggler A (WA), and Wiggler B (WB).

  11. Fabrication of the APS Storage Ring radio frequency accelerating cavities

    SciTech Connect

    Primdahl, K.; Bridges, J.; DePaola, F.; Kustom, R.; Snee, D.

    1993-07-01

    Specification, heat treatment, strength, and fatigue life of the Advanced Photon Source (APS) Storage Ring 352-MHz radio frequency (RF) accelerating cavity copper is discussed. Heat transfer studies, including finite element analysis, and configuration of water cooling is described. Requirements for and techniques of machining are considered. Braze and electron beam joint designs are compared. Vacuum considerations during fabrication are discussed.

  12. ACTIVATION OF AP-1 IN UROTSA CELLS BY METHYLATED ARSENICALS

    EPA Science Inventory

    ACTIVATION OF AP-1 IN UROTSA CELLS BY METHYLATED TRIVALENT ARSENICALS. Z Drobna1, I Jaspers2, D J Thomas3 and M Styblo1. 1Department of Pediatrics; 2Center for Environmental Medicine and Lung Biology, University of North Carolina at Chapel Hill, NC, USA; 3US EPA, RTP, NC, USA.

  13. The AP Lever for Boosting Access, Success, and Equity

    ERIC Educational Resources Information Center

    Roegman, Rachel; Hatch, Thomas

    2016-01-01

    Four New Jersey school districts worked together to increase student achievement by applying a number of strategies focused on getting traditionally underrepresented students to take more AP courses. The districts are members of the New Jersey Network of Superintendents (NJNS), comprising 15 superintendents who work together to develop systemwide…

  14. Nuclear Reactor Safety--The APS Submits its Report

    ERIC Educational Resources Information Center

    Physics Today, 1975

    1975-01-01

    Presents the summary section of the American Physical Society (APS) report on the safety features of the light-water reactor, reviews the design, construction, and operation of a reactor and outlines the primary engineered safety features. Summarizes the major recommendations of the study group. (GS)

  15. Approximate entropy (ApEn) as a complexity measure

    NASA Astrophysics Data System (ADS)

    Pincus, Steve

    1995-03-01

    Approximate entropy (ApEn) is a recently developed statistic quantifying regularity and complexity, which appears to have potential application to a wide variety of relatively short (greater than 100 points) and noisy time-series data. The development of ApEn was motivated by data length constraints commonly encountered, e.g., in heart rate, EEG, and endocrine hormone secretion data sets. We describe ApEn implementation and interpretation, indicating its utility to distinguish correlated stochastic processes, and composite deterministic/ stochastic models. We discuss the key technical idea that motivates ApEn, that one need not fully reconstruct an attractor to discriminate in a statistically valid manner—marginal probability distributions often suffice for this purpose. Finally, we discuss why algorithms to compute, e.g., correlation dimension and the Kolmogorov-Sinai (KS) entropy, often work well for true dynamical systems, yet sometimes operationally confound for general models, with the aid of visual representations of reconstructed dynamics for two contrasting processes.

  16. Activation of transcription factor AP-2 mediates UVA radiation- and singlet oxygen-induced expression of the human intercellular adhesion molecule 1 gene

    SciTech Connect

    Grether-Beck, S.; Olaizola-Horn, S.; Schmitt, H.; Grewe, M.

    1996-12-10

    UVA radiation is the major component of the UV solar spectrum that reaches the earth, and the therapeutic application of UVA radiation is increasing in medicine. Analysis of the cellular effects of UVA radiation has revealed that exposure of human cells to UVA radiation at physiological doses leads to increased gene expression and that this UVA response is primarily mediated through the generation of singlet oxygen. In this study, the mechanisms by which UVA radiation induces transcriptional activation of the human intercellular adhesion molecule 1 (ICAM-1) were examined. UVA radiation was capable of inducing activation of the human ICAM-1 promoter and increasing OCAM-1 mRNA and protein expression. These UVA radiation effects were inhibited by singlet oxygen quenchers, augmented by enhancement of singlet oxygen life-time, and mimicked in unirradiated cells by a singlet oxygen-generating system. UVA radiation as well as singlet oxygen-induced ICAM-1 promoter activation required activation of the transcription factor AP-2. Accordingly, both stimuli activated AP-2, and deletion of the putative AP-2-binding site abrogated ICAM-1 promoter activation in this system. This study identified the AP-2 site as the UVA radiation- and singlet oxygen-responsive element of the human ICAM-1 gene. The capacity of UVA radiation and/or singlet oxygen to induce human gene expression through activation of AP-2 indicates a previously unrecognized role of this transcription factor in the mammalian stress response. 38 refs., 3 figs., 3 tabs.

  17. aP2-Cre-mediated inactivation of acetyl-CoA carboxylase 1 causes growth retardation and reduced lipid accumulation in adipose tissues

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Adipose tissue is one of the major sites for fatty acid synthesis and lipid storage. We generated adipose (fat)-specific ACC1 knockout (FACC1KO) mice using the aP2-Cre/loxP system. FACC1KO mice showed prenatal growth retardation; after weaning, however, their weight gain was comparable to that of wi...

  18. Identification of GATA2 and AP-1 activator elements within the enhancer VNTR occurring in intron 5 of the human SIRT3 gene

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Human SIRT3 gene contains an intronic VNTR enhancer. A T > C transition occurring in the second repeat of each VNTR allele implies the presence/absence of a putative GATA binding motif. A partially overlapping AP-1 site, not affected by the transition, was also identified. Aims of the present study ...

  19. Plant resistance against the parasitic nematode Heterodera schachtii is mediated by MPK3 and MPK6 kinases, which are controlled by the MAPK phosphatase AP2C1 in Arabidopsis

    PubMed Central

    Sidonskaya, Ekaterina; Schweighofer, Alois; Shubchynskyy, Volodymyr; Kammerhofer, Nina; Hofmann, Julia; Wieczorek, Krzysztof; Meskiene, Irute

    2016-01-01

    Plant-parasitic cyst nematodes infect plants and form highly sophisticated feeding sites in roots. It is not known which plant cell signalling mechanisms trigger plant defence during the early stages of nematode parasitism. Mitogen-activated protein kinases (MAPKs) are central components of protein phosphorylation cascades transducing extracellular signals to plant defence responses. MAPK phosphatases control kinase activities and the signalling outcome. The involvement and the role of MPK3 and MPK6, as well as the MAPK phosphatase AP2C1, is demonstrated during parasitism of the beet cyst nematode Heterodera schachtii in Arabidopsis. Our data reveal notable activation patterns of plant MAPKs and the induction of AP2C1 suggesting the attenuation of defence signalling in plant cells during early nematode infection. It is demonstrated that the ap2c1 mutant that is lacking AP2C1 is more attractive but less susceptible to nematodes compared with the AP2C1-overexpressing line. This implies that the function of AP2C1 is a negative regulator of nematode-induced defence. By contrast, the enhanced susceptibility of mpk3 and mpk6 plants indicates a positive role of stress-activated MAPKs in plant immunity against nematodes. Evidence is provided that phosphatase AP2C1, as well as AP2C1-targeted MPK3 and MPK6, are important regulators of plant–nematode interaction, where the co-ordinated action of these signalling components ensures the timely activation of plant defence. PMID:26438412

  20. Plant resistance against the parasitic nematode Heterodera schachtii is mediated by MPK3 and MPK6 kinases, which are controlled by the MAPK phosphatase AP2C1 in Arabidopsis.

    PubMed

    Sidonskaya, Ekaterina; Schweighofer, Alois; Shubchynskyy, Volodymyr; Kammerhofer, Nina; Hofmann, Julia; Wieczorek, Krzysztof; Meskiene, Irute

    2016-01-01

    Plant-parasitic cyst nematodes infect plants and form highly sophisticated feeding sites in roots. It is not known which plant cell signalling mechanisms trigger plant defence during the early stages of nematode parasitism. Mitogen-activated protein kinases (MAPKs) are central components of protein phosphorylation cascades transducing extracellular signals to plant defence responses. MAPK phosphatases control kinase activities and the signalling outcome. The involvement and the role of MPK3 and MPK6, as well as the MAPK phosphatase AP2C1, is demonstrated during parasitism of the beet cyst nematode Heterodera schachtii in Arabidopsis. Our data reveal notable activation patterns of plant MAPKs and the induction of AP2C1 suggesting the attenuation of defence signalling in plant cells during early nematode infection. It is demonstrated that the ap2c1 mutant that is lacking AP2C1 is more attractive but less susceptible to nematodes compared with the AP2C1-overexpressing line. This implies that the function of AP2C1 is a negative regulator of nematode-induced defence. By contrast, the enhanced susceptibility of mpk3 and mpk6 plants indicates a positive role of stress-activated MAPKs in plant immunity against nematodes. Evidence is provided that phosphatase AP2C1, as well as AP2C1-targeted MPK3 and MPK6, are important regulators of plant-nematode interaction, where the co-ordinated action of these signalling components ensures the timely activation of plant defence. PMID:26438412

  1. Signalling in inflammatory skin disease by AP-1 (Fos/Jun).

    PubMed

    Uluçkan, Özge; Guinea-Viniegra, Juan; Jimenez, Maria; Wagner, Erwin F

    2015-01-01

    Skin inflammation is a physiological reaction to tissue injury, pathogen invasion and irritants. During this process, innate and/or adaptive immune cells are activated and recruited to the site of inflammation to either promote or suppress inflammation. The sequential recruitment and activation of immune cells is modulated by a combination of cytokines and chemokines, which are regulated by transcription factors, such as AP-1 (Fos/Jun), NF-κB, NFATs, and STATs. Here we review the present evidence and the underlying mechanisms of how Jun/AP-1 proteins control skin inflammation. Genetically engineered mouse models (GEMMs) in which AP-1 proteins are deleted in the epidermis have revealed that these proteins control cytokine expression at multiple levels. Constitutive epidermal deletion of JunB in mice leads to a multi-organ disease characterised by increased levels of pro-inflammatory cytokines. These JunB-deficient mutant mice display several phenotypes from skin inflammation to a G-CSF-dependent myeloproliferative disease, as well as kidney atrophy and bone loss, reminiscent of psoriasis and systemic lupus erythematosus. Importantly, epidermal deletion of both JunB and c-Jun in an inducible manner in adult mice leads to a psoriasis-like disease, in which the epidermal proteome expression profile is comparable to the one from psoriasis patient samples. In this GEMM and in psoriasis patient-derived material, S100A8/A9-dependent C3/CFB complement activation, as well as a miR-21-dependent TIMP-3/TACE pathway leading to TNF-α shedding, plays causal roles in disease development. The newly identified therapeutic targets from GEMMs together with investigations in human patient samples open up new avenues for therapeutic interventions for psoriasis and related inflammatory skin diseases. PMID:26458100

  2. Aviation Safety Program: Weather Accident Prevention (WxAP) Development of WxAP System Architecture And Concepts of Operation

    NASA Technical Reports Server (NTRS)

    Grantier, David

    2003-01-01

    This paper presents viewgraphs on the development of the Weather Accident Prevention (WxAP) System architecture and Concept of Operation (CONOPS) activities. The topics include: 1) Background Information on System Architecture/CONOPS Activity; 2) Activity Work in Progress; and 3) Anticipated By-Products.

  3. Transcriptional activation of the fra-1 gene by AP-1 is mediated by regulatory sequences in the first intron.

    PubMed Central

    Bergers, G; Graninger, P; Braselmann, S; Wrighton, C; Busslinger, M

    1995-01-01

    Constitutive expression of c-Fos, FosB, Fra-1, or c-Jun in rat fibroblasts leads to up-regulation of the immediate-early gene fra-1. Using the posttranslational FosER induction system, we demonstrate that this AP-1-dependent stimulation of fra-1 expression is rapid, depends on a functional DNA-binding domain of FosER, and is a general phenomenon observed in different cell types. In vitro mutagenesis and functional analysis of the rat fra-1 gene in stably transfected Rat-1A-FosER fibroblasts indicated that basal and AP-1-regulated expression of the fra-1 gene depends on regulatory sequences in the first intron which comprise a consensus AP-1 site and two AP-1-like elements. We have also investigated the transactivating and transforming properties of the Fra-1 protein to address the significance of fra-1 up-regulation. The entire Fra-1 protein fused to the DNA-binding domain of Ga14 is shown to lack any transactivation function, and yet it possesses oncogenic potential, as overexpression of Fra-1 in established rat fibroblasts results in anchorage-independent growth in vitro and tumor development in athymic mice, fra-1 is therefore not only induced by members of the Fos family, but its gene product may also contribute to cellular transformation by these proteins. Together, these data identify fra-1 as a unique member of the fos gene family which is under positive control by AP-1 activity. PMID:7791782

  4. A Common Signal Patch Drives AP-1 Protein-dependent Golgi Export of Inwardly Rectifying Potassium Channels.

    PubMed

    Li, Xiangming; Ortega, Bernardo; Kim, Boyoung; Welling, Paul A

    2016-07-15

    Nearly all members of the inwardly rectifying potassium (Kir) channel family share a cytoplasmic domain structure that serves as an unusual AP-1 clathrin adaptor-dependent Golgi export signal in one Kir channel, Kir2.1 (KCNJ2), raising the question whether Kir channels share a common Golgi export mechanism. Here we explore this idea, focusing on two structurally and functionally divergent Kir family members, Kir2.3 (KCNJ4) and Kir4.1/5.1 (KCNJ10/16), which have ∼50% amino identity. We found that Golgi export of both channels is blocked upon siRNA-mediated knockdown of the AP-1 γ subunit, as predicted for the common AP-1-dependent trafficking process. A comprehensive mutagenic analysis, guided by homology mapping in atomic resolution models of Kir2.1, Kir2.3, and Kir4.1/5.1, identified a common structure that serves as a recognition site for AP-1 binding and governs Golgi export. Larger than realized from previous studies with Kir2.1, the signal is created by a patch of residues distributed at the confluence of cytoplasmic N and C termini. The signal involves a stretch of hydrophobic residues from the C-terminal region that form a hydrophobic cleft, an adjacent cluster of basic residues within the N terminus, and a potential network of salt bridges that join the N- and C-terminal poles together. Because patch formation and AP-1 binding are dependent on proper folding of the cytoplasmic domains, the signal provides a common quality control mechanism at the Golgi for Kir channels. These findings identify a new proteostatic mechanism that couples protein folding of channels to forward trafficking in the secretory pathway. PMID:27226616

  5. The p38 SAPK pathway regulates the expression of the MMP-9 collagenase via AP-1-dependent promoter activation.

    PubMed

    Simon, C; Simon, M; Vucelic, G; Hicks, M J; Plinkert, P K; Koitschev, A; Zenner, H P

    2001-12-10

    The invasive phenotype of cancers critically depends on the expression of proteases such as the M(R) 92,000 type IV collagenase (MMP-9). Several growth factors and oncogenes were found to increase promoter activity and as a consequence protease expression. This frequently requires the activation of the transcription factor AP-1 by signal transduction cascades such as the ERK and JNK pathways. We have previously demonstrated that the tumor promoter TPA can induce MMP-9 expression via a third signaling cascade, the p38 pathway. Considering that TPA is a potent activator of AP-1, we hypothesized that this transcription factor might also be required for p38 pathway-dependent MMP-9 regulation. While dominant negative p38 and MKK-6 mutants reduced MMP-9 promoter activity in CAT assays, a construct encoding an activating mutation in the MKK-6 protein potently stimulated it. This was mediated via 144 bp of the 5'flanking region of the wild-type promoter, which contains an AP-1 site at -79. Both point mutations in this motif and the expression of a c-jun protein lacking its transactivation domain and therefore acting as a dominant negative AP-1 mutant abrogated MKK-6-dependent promoter stimulation. Finally SB 203580, a specific p38 pathway inhibitor, reduced MMP-9 expression/secretion and in vitro invasion of cancer cells. Thus, our results provide evidence that also the third SAPK/MAPK signaling cascade, the p38 signal transduction pathway, stimulates MMP-9 expression in an AP-1-dependent fashion. PMID:11716547

  6. NF-κB and AP-1 Connection: Mechanism of NF-κB-Dependent Regulation of AP-1 Activity

    PubMed Central

    Fujioka, Shuichi; Niu, Jiangong; Schmidt, Christian; Sclabas, Guido M.; Peng, Bailu; Uwagawa, Tadashi; Li, Zhongkui; Evans, Douglas B.; Abbruzzese, James L.; Chiao, Paul J.

    2004-01-01

    Nuclear factor κB (NF-κB) and activator protein 1 (AP-1) transcription factors regulate many important biological and pathological processes. Activation of NF-κB is regulated by the inducible phosphorylation of NF-κB inhibitor IκB by IκB kinase. In contrast, Fos, a key component of AP-1, is primarily transcriptionally regulated by serum responsive factors (SRFs) and ternary complex factors (TCFs). Despite these different regulatory mechanisms, there is an intriguing possibility that NF-κB and AP-1 may modulate each other, thus expanding the scope of these two rapidly inducible transcription factors. To determine whether NF-κB activity is involved in the regulation of fos expression in response to various stimuli, we analyzed activity of AP-1 and expression of fos, fosB, fra-1, fra-2, jun, junB, and junD, as well as AP-1 downstream target gene VEGF, using MDAPanc-28 and MDAPanc-28/IκBαM pancreatic tumor cells and wild-type, IKK1−/−, and IKK2−/− murine embryonic fibroblast cells. Our results show that elk-1, a member of TCFs, is one of the NF-κB downstream target genes. Inhibition of NF-κB activity greatly decreased expression of elk-1. Consequently, the reduced level of activated Elk-1 protein by extracellular signal-regulated kinase impeded constitutive, serum-, and superoxide-inducible c-fos expression. Thus, our study revealed a distinct and essential role of NF-κB in participating in the regulation of elk-1, c-fos, and VEGF expression. PMID:15314185

  7. Heparin (GAG-hed) inhibits LCR activity of Human Papillomavirus type 18 by decreasing AP1 binding

    PubMed Central

    Villanueva, Rita; Morales-Peza, Néstor; Castelán-Sánchez, Irma; García-Villa, Enrique; Tapia, Rocio; Cid-Arregui, Ángel; García-Carrancá, Alejandro; López-Bayghen, Esther; Gariglio, Patricio

    2006-01-01

    Background High risk HPVs are causative agents of anogenital cancers. Viral E6 and E7 genes are continuously expressed and are largely responsible for the oncogenic activity of these viruses. Transcription of the E6 and E7 genes is controlled by the viral Long Control Region (LCR), plus several cellular transcription factors including AP1 and the viral protein E2. Within the LCR, the binding and activity of the transcription factor AP1 represents a key regulatory event in maintaining E6/E7 gene expression and uncontrolled cell proliferation. Glycosaminoglycans (GAGs), such as heparin, can inhibit tumour growth; they have also shown antiviral effects and inhibition of AP1 transcriptional activity. The purpose of this study was to test the heparinoid GAG-hed, as a possible antiviral and antitumoral agent in an HPV18 positive HeLa cell line. Methods Using in vivo and in vitro approaches we tested GAG-hed effects on HeLa tumour cell growth, cell proliferation and on the expression of HPV18 E6/E7 oncogenes. GAG-hed effects on AP1 binding to HPV18-LCR-DNA were tested by EMSA. Results We were able to record the antitumoral effect of GAG-hed in vivo by using as a model tumours induced by injection of HeLa cells into athymic female mice. The antiviral effect of GAG-hed resulted in the inhibition of LCR activity and, consequently, the inhibition of E6 and E7 transcription. A specific diminishing of cell proliferation rates was observed in HeLa but not in HPV-free colorectal adenocarcinoma cells. Treated HeLa cells did not undergo apoptosis but the percentage of cells in G2/M phase of the cell cycle was increased. We also detected that GAG-hed prevents the binding of the transcription factor AP1 to the LCR. Conclusion Direct interaction of GAG-hed with the components of the AP1 complex and subsequent interference with its ability to correctly bind specific sites within the viral LCR may contribute to the inhibition of E6/E7 transcription and cell proliferation. Our data

  8. DNA damage processing by human 8-oxoguanine-DNA glycosylase mutants with the occluded active site.

    PubMed

    Lukina, Maria V; Popov, Alexander V; Koval, Vladimir V; Vorobjev, Yuri N; Fedorova, Olga S; Zharkov, Dmitry O

    2013-10-01

    8-Oxoguanine-DNA glycosylase (OGG1) removes premutagenic lesion 8-oxoguanine (8-oxo-G) from DNA and then nicks the nascent abasic (apurinic/apyrimidinic) site by β-elimination. Although the structure of OGG1 bound to damaged DNA is known, the dynamic aspects of 8-oxo-G recognition are not well understood. To comprehend the mechanisms of substrate recognition and processing, we have constructed OGG1 mutants with the active site occluded by replacement of Cys-253, which forms a wall of the base-binding pocket, with bulky leucine or isoleucine. The conformational dynamics of OGG1 mutants were characterized by single-turnover kinetics and stopped-flow kinetics with fluorescent detection. Additionally, the conformational mobility of wild type and the mutant OGG1 substrate complex was assessed using molecular dynamics simulations. Although pocket occlusion distorted the active site and greatly decreased the catalytic activity of OGG1, it did not fully prevent processing of 8-oxo-G and apurinic/apyrimidinic sites. Both mutants were notably stimulated in the presence of free 8-bromoguanine, indicating that this base can bind to the distorted OGG1 and facilitate β-elimination. The results agree with the concept of enzyme plasticity, suggesting that the active site of OGG1 is flexible enough to compensate partially for distortions caused by mutation. PMID:23955443

  9. Distinct and separable activities of the endocytic clathrin coat components Fcho1/2 and AP-2 in developmental patterning

    PubMed Central

    Umasankar, P. K.; Sanker, Subramaniam; Thieman, James R.; Chakraborty, Souvik; Wendland, Beverly; Tsang, Michael; Traub, Linton M.

    2012-01-01

    Clathrin-mediated endocytosis occurs at multiple independent import sites on the plasma membrane, but how these positions are selected and how different cargo is simultaneously recognized is obscure. FCHO1 and FCHO2 are early-arriving proteins at surface clathrin assemblies and are speculated to act as compulsory coat nucleators, preceding the core clathrin adaptor AP-2. Here, we show the μ-homology domain (μHD) of FCHO1/2 represents a novel endocytic interaction hub. Translational silencing of fcho1 in zebrafish embryos causes strong dorsoventral patterning defects analogous to Bmp signal failure. The Fcho1 μHD interacts with the Bmp receptor Alk8, uncovering a new endocytic component that positively modulates Bmp signal transmission. Still, the fcho1 morphant phenotype is distinct from severe embryonic defects apparent when AP-2 is depleted. Our data thus contradict the primacy of FCHO1/2 in coat initiation. PMID:22484487

  10. Elastic Coupling of Nascent apCAM Adhesions to Flowing Actin Networks

    PubMed Central

    Mejean, Cecile O.; Schaefer, Andrew W.; Buck, Kenneth B.; Kress, Holger; Shundrovsky, Alla; Merrill, Jason W.; Dufresne, Eric R.; Forscher, Paul

    2013-01-01

    Adhesions are multi-molecular complexes that transmit forces generated by a cell’s acto-myosin networks to external substrates. While the physical properties of some of the individual components of adhesions have been carefully characterized, the mechanics of the coupling between the cytoskeleton and the adhesion site as a whole are just beginning to be revealed. We characterized the mechanics of nascent adhesions mediated by the immunoglobulin-family cell adhesion molecule apCAM, which is known to interact with actin filaments. Using simultaneous visualization of actin flow and quantification of forces transmitted to apCAM-coated beads restrained with an optical trap, we found that adhesions are dynamic structures capable of transmitting a wide range of forces. For forces in the picoNewton scale, the nascent adhesions’ mechanical properties are dominated by an elastic structure which can be reversibly deformed by up to 1 µm. Large reversible deformations rule out an interface between substrate and cytoskeleton that is dominated by a number of stiff molecular springs in parallel, and favor a compliant cross-linked network. Such a compliant structure may increase the lifetime of a nascent adhesion, facilitating signaling and reinforcement. PMID:24039928

  11. Airborne Precision Spacing (APS) Dependent Parallel Arrivals (DPA)

    NASA Technical Reports Server (NTRS)

    Smith, Colin L.

    2012-01-01

    The Airborne Precision Spacing (APS) team at the NASA Langley Research Center (LaRC) has been developing a concept of operations to extend the current APS concept to support dependent approaches to parallel or converging runways along with the required pilot and controller procedures and pilot interfaces. A staggered operations capability for the Airborne Spacing for Terminal Arrival Routes (ASTAR) tool was developed and designated as ASTAR10. ASTAR10 has reached a sufficient level of maturity to be validated and tested through a fast-time simulation. The purpose of the experiment was to identify and resolve any remaining issues in the ASTAR10 algorithm, as well as put the concept of operations through a practical test.

  12. The APS booster synchrotron: Commissioning and operational experience

    SciTech Connect

    Milton, S.V.

    1995-07-01

    The Advanced Photon Source (APS) at Argonne National Laboratory (ANL) was constructed to provide a large user community with intense and high brightness synchrotron radiation at x-ray wavelengths. A 7-GeV positron beam is used to generate this light. Acceleration of the beam from 450 MeV to 7 GeV is accomplished at a 2-Hz repetition rate by the booster synchrotron. Commissioning of the booster began in the second quarter of 1994 and continued on into early 1995. The booster is now routinely used to provide beam for the commissioning of the APS storage ring. Reported here are our commissioning and operational experiences with the booster synchrotron.

  13. Tank 241-AP-107 tank characterization plan. Revision 1

    SciTech Connect

    Schreiber, R.D.

    1995-01-20

    Defense Nuclear Facilities Safety Board has directed the DOE to concentrate ear-term sampling and analysis activities on identification and resolution of issues (Conway 1993). The Data Quality Objective (DQO) process was chosen as a tool to be used in the resolution of safety issues. As a result, a revision in the Federal Facilities Agreement and Consent Order (Tri-Party Agreement) milestone M-44-00 has been made, which states that ``A Tank Characterization Plan (TCP) will be developed for each double-shell tank (DST) and single-shell tank (SST) using the DQO process; Development of TCPs by the DQO process is intended to allow users (e.g., Hanford Facility user groups, regulators) to ensure their needs will be met and that resources are devoted to gaining only necessary information.`` This document satisfies that requirement for the tank 241-AP-107 (AP-107).

  14. Introduction to Physics of the Universe in AP Physics Classrooms

    NASA Astrophysics Data System (ADS)

    Allen, Stephanie L.

    2006-12-01

    Often students have difficulty understanding the connections that must be made between old and new concepts. This curriculum is designed to lead AP Physics students through this process with gamma ray bursts. Students will participate in various discussions, demonstrations, exercises and activities that lead them through universe basics, life cycles of stars, black holes, the electromagnetic spectrum, and they will learn about various NASA observatories. Ultimately, this will result in a better understanding of how astrophysicists have come to understand the gamma ray burst. This series of lessons was created for the three to four weeks after the AP Physics exam. The curriculum was developed through gathering resources from various scientific organizations, developing new ideas and speaking with scientists at NASA Goddard Space Flight Center. The result is a manual of lesson plans, activities and answer keys for teachers to use in their own classrooms.

  15. Variability of Balmer Profiles in Magnetic Ap/Bp Stars

    NASA Astrophysics Data System (ADS)

    Valyavin, G.; Lee, B.-C.; Shulyak, D.; Han, I.; Kochukhov, O.; Khang, D.-I.; Kim, K.-M.

    2007-06-01

    A set of high precision measurements of weak variations of hydrogen lines in spectra of seven magnetic Ap/Bp stars was carried out using the BOES echelle spectrograph of the Bohuynsan Optical Astronomy Observatory (South Korea). A weak (1-2 %) periodic variability of the Balmer line wings has been detected in the spectra of 2 program stars. Upper limits of possible variations are presented for the remaining 5 objects. We discuss the discovered variability in the framework of model atmospheres with magnetic force terms included. The periodic changes in the Balmer profiles are caused by perturbations in atmospheres of Ap/Bp stars due to their rotationally modulated non-force-free magnetic fields.

  16. Measurements of ground motion and magnet vibrations at the APS

    SciTech Connect

    Shiltsev, V.

    1996-09-01

    This article presents results of ground motion and magnet vibrations measurements at the Advanced Photon Source. The experiments were done over a wide, frequency range (0-05-100 Hz) with the use of SM-3KV-type seismic probes from the Budker Institute of Nuclear Physics (Russia). Spectral power densities of vertical and horizontal motions of the APS hall floor and quadrupoles on regular supports were obtained. Also investigated were magnet vibrations induced by designed cooling water flow and spectral characteristics of spatial correlation of the quadrupole vibrations at different sectors of the ring. The influence of personnel activity in the hall and traffic under the ring on the slow motion of storage ring elements were observed. Amplitudes of vibrations at the APS are compared with results of seismic measurements at some other accelerators.

  17. Collagen XVI Induces Expression of MMP9 via Modulation of AP-1 Transcription Factors and Facilitates Invasion of Oral Squamous Cell Carcinoma

    PubMed Central

    Bedal, Konstanze B.; Grässel, Susanne; Oefner, Peter J.; Reinders, Joerg; Reichert, Torsten E.; Bauer, Richard

    2014-01-01

    Collagen XVI belongs to the family of fibril-associated collagens with interrupted triple helices (FACIT). It is overexpressed during the progression of oral squamous cell carcinoma (OSCC). The present data show a strong collagen XVI-dependent induction of MMP9 and an increase in OSCC cell invasion. We found activated integrin-linked kinase (ILK) in a complex with kindlin-1 and activation of protein kinase B (PKB/Akt) to be responsible for MMP9 induction. Inhibition of the formation of focal adhesions reduced MMP9 expression. Moreover, collagen XVI overexpressing OSCC cell clones (COLXVI cell clones) transfected with vectors containing different MMP9 promoter fragments adjacent to a luciferase reporter revealed an increase in luciferase signal dependent on AP-1 binding sites. Deletion of the AP-1 binding site 98 bp upstream of the reported transcription start site and inhibition of AP-1 with Tanshinone IIA resulted in decreased MMP9 expression. The AP-1 subunit JunB showed differential expression between COLXVI cell clones and mock control cells. Additionally, mass spectrometric analysis of immunoprecipitates revealed that c-Fos interacted strongly with dyskerin in COLXVI cell clones compared to mock controls. PMID:24466237

  18. QCD on the highly parallel computer AP1000

    NASA Astrophysics Data System (ADS)

    Akemi, K.; de Forcrand, Ph.; Fujisaki, M.; Hashimoto, T.; Hege, H. C.; Hioki, S.; Makino, J.; Miyamura, O.; Nakamura, A.; Okuda, M.; Stamatescu, I. O.; Tago, Y.; Takaishi, T.; QCD TARO (QCD on Thousand cell ARray processorsOmnipurpose) Collaboration

    We have been running quenched QCD simulations on 32 4 and 32 3 × 48 lattices using a 512 processor AP1000, which is a highly parallel computer with up to 1024 processing elements. We have developed programs for update, blocking and hadron propagator calculations. The pseudo heatbath and the overrelaxation algorithms were used for the update with performance of 2.6 and 2.0 μsec/link, respectively.

  19. Diffusion and Settling in Ap/Bp Stars

    SciTech Connect

    Turcotte, S

    2003-04-09

    Ap/Bp stars are magnetic chemically peculiar early A and late B type stars of the main sequence. They exhibit peculiar surface abundance anomalies that are thought to be the result of gravitational settling and radiative levitation. The physics of diffusion in these stars are reviewed briefly and some model predictions are discussed. While models reproduce some observations reasonably well, more work is needed before the behavior of diffusing elements in a complex magnetic field is fully understood.

  20. Architecture of the APS real-time orbit feedback system.

    SciTech Connect

    Carwardine, J. A.; Lenkszus, F. R.

    1997-11-21

    The APS Real-Time Orbit Feedback System is designed to stabilize the orbit of the stored positron beam against low-frequency sources such as mechanical vibration and power supply ripple. A distributed array of digital signal processors is used to measure the orbit and compute corrections at a 1kHz rate. The system also provides extensive beam diagnostic tools. This paper describes the architectural aspects of the system and describes how the orbit correction algorithms are implemented.

  1. Identification and targeting of an interaction between a tyrosine motif within hepatitis C virus core protein and AP2M1 essential for viral assembly.

    PubMed

    Neveu, Gregory; Barouch-Bentov, Rina; Ziv-Av, Amotz; Gerber, Doron; Jacob, Yves; Einav, Shirit

    2012-01-01

    Novel therapies are urgently needed against hepatitis C virus infection (HCV), a major global health problem. The current model of infectious virus production suggests that HCV virions are assembled on or near the surface of lipid droplets, acquire their envelope at the ER, and egress through the secretory pathway. The mechanisms of HCV assembly and particularly the role of viral-host protein-protein interactions in mediating this process are, however, poorly understood. We identified a conserved heretofore unrecognized YXXΦ motif (Φ is a bulky hydrophobic residue) within the core protein. This motif is homologous to sorting signals within host cargo proteins known to mediate binding of AP2M1, the μ subunit of clathrin adaptor protein complex 2 (AP-2), and intracellular trafficking. Using microfluidics affinity analysis, protein-fragment complementation assays, and co-immunoprecipitations in infected cells, we show that this motif mediates core binding to AP2M1. YXXΦ mutations, silencing AP2M1 expression or overexpressing a dominant negative AP2M1 mutant had no effect on HCV RNA replication, however, they dramatically inhibited intra- and extracellular infectivity, consistent with a defect in viral assembly. Quantitative confocal immunofluorescence analysis revealed that core's YXXΦ motif mediates recruitment of AP2M1 to lipid droplets and that the observed defect in HCV assembly following disruption of core-AP2M1 binding correlates with accumulation of core on lipid droplets, reduced core colocalization with E2 and reduced core localization to trans-Golgi network (TGN), the presumed site of viral particles maturation. Furthermore, AAK1 and GAK, serine/threonine kinases known to stimulate binding of AP2M1 to host cargo proteins, regulate core-AP2M1 binding and are essential for HCV assembly. Last, approved anti-cancer drugs that inhibit AAK1 or GAK not only disrupt core-AP2M1 binding, but also significantly inhibit HCV assembly and infectious virus production

  2. Human kidney anion exchanger 1 interacts with adaptor-related protein complex 1 {mu}1A (AP-1 mu1A)

    SciTech Connect

    Sawasdee, Nunghathai; Junking, Mutita; Ngaojanlar, Piengpaga; Sukomon, Nattakan; Ungsupravate, Duangporn; Limjindaporn, Thawornchai; Akkarapatumwong, Varaporn; Noisakran, Sansanee; Yenchitsomanus, Pa-thai

    2010-10-08

    Research highlights: {yields} Trafficking defect of kAE1 is a cause of dRTA but trafficking pathway of kAE1 has not been clearly described. {yields} Adaptor-related protein complex 1 {mu}1A (AP-1 mu1A) was firstly reported to interact with kAE1. {yields} The interacting site for AP-1 mu1A on Ct-kAE1 was found to be Y904DEV907, a subset of YXXO motif. {yields} AP-1 mu1A knockdown showed a marked reduction of kAE1 on the cell membrane and its accumulation in endoplasmic reticulum. {yields} AP-1 mu1A has a critical role in kAE1 trafficking to the plasma membrane. -- Abstract: Kidney anion exchanger 1 (kAE1) mediates chloride (Cl{sup -}) and bicarbonate (HCO{sub 3}{sup -}) exchange at the basolateral membrane of kidney {alpha}-intercalated cells. Impaired trafficking of kAE1 leads to defect of the Cl{sup -}/HCO{sub 3}{sup -} exchange at the basolateral membrane and failure of proton (H{sup +}) secretion at the apical membrane, causing a kidney disease - distal renal tubular acidosis (dRTA). To gain a better insight into kAE1 trafficking, we searched for proteins physically interacting with the C-terminal region of kAE1 (Ct-kAE1), which contains motifs crucial for intracellular trafficking, by a yeast two-hybrid (Y2H) system. An adaptor-related protein complex 1 {mu}1A (AP-1 mu1A) subunit was found to interact with Ct-kAE1. The interaction between either Ct-kAE1 or full-length kAE1 and AP-1 mu1A were confirmed in human embryonic kidney (HEK) 293T by co-immunoprecipitation, affinity co-purification, co-localization, yellow fluorescent protein (YFP)-based protein fragment complementation assay (PCA) and GST pull-down assay. The interacting site for AP-1 mu1A on Ct-kAE1 was found to be Y904DEV907, a subset of YXXO motif. Interestingly, suppression of endogenous AP-1 mu1A in HEK 293T by small interfering RNA (siRNA) decreased membrane localization of kAE1 and increased its intracellular accumulation, suggesting for the first time that AP-1 mu1A is involved in the kAE1

  3. Margin Assessment of AP1000 Loss of Flow Transient

    SciTech Connect

    Carlin, Edward L.; Hilton, Peter A.; Yixing Sung

    2006-07-01

    The Reactor Coolant System (RCS) of the AP1000 plant consists of two circulating loops. Each loop contains two canned motor Reactor Coolant (RC) pumps that have a rotating inertia to provide RCS flow coast-down if power to the pumps is lost. Westinghouse analysis of the complete loss of flow (CLOF) accident in support of the AP1000 design certification was based on the USNRC-approved traditional methodology applied to operating plants. The RCS response during the transient was predicted using the LOFTRAN code based on a reactivity insertion curve highly skewed to the bottom of the reactor core, but the calculation of Departure from Nucleate Boiling Ratio (DNBR) was performed assuming a top-skewed axial power profile. A more realistic margin assessment can be made by using an improved method similar to Westinghouse RAVE methodology recently approved by the USNRC. The improved method uses the three-dimensional kinetic nodal code SPNOVA coupled with the reactor core thermal-hydraulic code VIPRE-W for predicting the reactor core response during the CLOF transient. The improved method significantly improves margin predictions by generating core power distributions consistent with the trip reactivity changes for the DNBR calculation. The margin assessment showed that the improved method resulted in a 19% DNBR increase as compared to the traditional method for the AP1000 CLOF transient. (authors)

  4. Discoveries in the Atmospheres of roAp Stars

    NASA Astrophysics Data System (ADS)

    Kurtz, D. W.; Freyhammer, L. M.; Elkin, V. G.; Mathys, G.

    2007-11-01

    We have obtained a large amount of data on over 40 roAp stars and potential roAp stars with the Ultraviolet and Visual Echelle Spectrograph (UVES) on the VLT with time resolution typically around 1 min and radial velocity precision as high as 1 m s-1. Abundance stratification caused by atomic diffusion in the presence of strong global magnetic fields gives promise of three-dimensional maps of the pulsation amplitude and phase, and of the abundance distributions of many ions that may provide the strongest observational tests of atomic diffusion theory. Studies of individual spectral lines and of line profile variability sample the observable atmospheres of roAp stars from continuum optical depth τ5000~1 to as high as τ5000~10-5, revealing fascinating new pulsational behaviour not observed in other types of pulsating stars, including, inter alia, line profile variability in rare earth elements lines interpreted by as evidence for shock waves in the high atmosphere of these stars, an intriguing range of line bisector shapes, and a new pulsational diagnostic for resolved Zeeman components for the most strongly magnetic stars.

  5. A Semi-automated Abundance Survey of Ap Stars

    NASA Astrophysics Data System (ADS)

    Hall, Martin P.; Kurtz, Don; Elkin, Vladimir; Bruntt, Hans

    2015-08-01

    We have carried out an abundance analysis on the high-resolution spectra of approximately 350 Ap stars collected between 2007 and 2010 on the FEROS Echelle (Fibre-led, Extended Range, Echelle ) spectrograph housed at the 2.2-m telescope at European Southern Observatory at La Silla, Chile. We employed the VWA package (vsin I, wavelength shift, abundance analysis) for preliminary selection of spectral lines, and a semi-automated set of routines which we developed in the programming language IDL, to calculate the equivalent widths and abundances of ions of Iron and the rare earth elements Neodymium and Praseodymium using the WIDTH program and NEMO model atmospheres. Initial results are presented, which reinforce the correlation between iron abundance and effective temperature, from an over-abundance in the late Bp stars, to under-abundant in the early F stars. Results also suggest that the disequilibrium in abundances of the first and second ionisation stages of these ions in the rapidly oscillating Ap (roAp) stars may a consequence of the relatively cool temperatures of those stars, rather than a signature of pulsation.

  6. Maternal AP determinants in the Drosophila oocyte and embryo.

    PubMed

    Ma, Jun; He, Feng; Xie, Gengqiang; Deng, Wu-Min

    2016-09-01

    An animal embryo cannot initiate its journey of forming a new life on its own. It must rely on maternally provided resources and inputs to kick-start its developmental process. In Drosophila, the initial polarities of the embryo along both the anterior-posterior (AP) and dorsal-ventral (DV) axes are also specified by maternal determinants. Over the past several decades, genetic and molecular studies have identified and characterized such determinants, as well as the zygotic genetic regulatory networks that control patterning in the early embryo. Extensive studies of oogenesis have also led to a detailed knowledge of the cellular and molecular interactions that control the formation of a mature egg. Despite these efforts, oogenesis and embryogenesis have been studied largely as separate problems, except for qualitative aspects with regard to maternal regulation of the asymmetric localization of maternal determinants. Can oogenesis and embryogenesis be viewed from a unified perspective at a quantitative level, and can that improve our understanding of how robust embryonic patterning is achieved? Here, we discuss the basic knowledge of the regulatory mechanisms controlling oogenesis and embryonic patterning along the AP axis. We explore properties of the maternal Bicoid gradient in relation to embryo size in search for a unified framework for robust AP patterning. WIREs Dev Biol 2016, 5:562-581. doi: 10.1002/wdev.235 For further resources related to this article, please visit the WIREs website. PMID:27253156

  7. Structure of the E6/E6AP/p53 complex required for HPV-mediated degradation of p53

    PubMed Central

    Martinez-Zapien, Denise; Ruiz, Francesc Xavier; Poirson, Juline; Mitschler, André; Ramirez-Ramos, Juan; Forster, Anne; Cousido-Siah, Alexandra; Masson, Murielle; Pol, Scott Vande; Podjarny, Alberto; Travé, Gilles; Zanier, Katia

    2015-01-01

    Summary The p53 pro-apoptotic tumor suppressor is mutated or functionally altered in most cancers. In epithelial tumors induced by “high-risk” mucosal Human Papillomaviruses (hrm-HPVs), including human cervical carcinoma and a growing number of head-and-neck cancers 1, p53 is degraded by the viral oncoprotein E6 2. In this process, E6 binds to a short LxxLL consensus sequence within the cellular ubiquitin ligase E6AP 3. Subsequently, the E6/E6AP heterodimer recruits and degrades p53 4. Neither E6 nor E6AP are separately able to recruit p53 3,5, and the precise mode of assembly of E6, E6AP and p53 is unknown. Here, we solved the crystal structure of a ternary complex comprising full-length HPV16 E6, the LxxLL motif of E6AP and the core domain of p53. The LxxLL motif of E6AP renders the conformation of E6 competent for interaction with p53 by structuring a p53-binding cleft on E6. Mutagenesis of critical positions at the E6-p53 interface disrupts p53 degradation. The E6-binding site of p53 is distal from previously described DNA- and protein-binding surfaces of the core domain. This suggests that, in principle, E6 may avoid competition with cellular factors by targeting both free and bound p53 molecules. The E6/E6AP/p53 complex represents a prototype of viral hijacking of both the ubiquitin-mediated protein degradation pathway and the p53 tumor suppressor pathway. The present structure provides a framework for the design of inhibitory therapeutic strategies against HPV-mediated oncogenesis. PMID:26789255

  8. Structure of the E6/E6AP/p53 complex required for HPV-mediated degradation of p53.

    PubMed

    Martinez-Zapien, Denise; Ruiz, Francesc Xavier; Poirson, Juline; Mitschler, André; Ramirez, Juan; Forster, Anne; Cousido-Siah, Alexandra; Masson, Murielle; Vande Pol, Scott; Podjarny, Alberto; Travé, Gilles; Zanier, Katia

    2016-01-28

    The p53 pro-apoptotic tumour suppressor is mutated or functionally altered in most cancers. In epithelial tumours induced by 'high-risk' mucosal human papilloma viruses, including human cervical carcinoma and a growing number of head-and-neck cancers, p53 is degraded by the viral oncoprotein E6 (ref. 2). In this process, E6 binds to a short leucine (L)-rich LxxLL consensus sequence within the cellular ubiquitin ligase E6AP. Subsequently, the E6/E6AP heterodimer recruits and degrades p53 (ref. 4). Neither E6 nor E6AP are separately able to recruit p53 (refs 3, 5), and the precise mode of assembly of E6, E6AP and p53 is unknown. Here we solve the crystal structure of a ternary complex comprising full-length human papilloma virus type 16 (HPV-16) E6, the LxxLL motif of E6AP and the core domain of p53. The LxxLL motif of E6AP renders the conformation of E6 competent for interaction with p53 by structuring a p53-binding cleft on E6. Mutagenesis of critical positions at the E6-p53 interface disrupts p53 degradation. The E6-binding site of p53 is distal from previously described DNA- and protein-binding surfaces of the core domain. This suggests that, in principle, E6 may avoid competition with cellular factors by targeting both free and bound p53 molecules. The E6/E6AP/p53 complex represents a prototype of viral hijacking of both the ubiquitin-mediated protein degradation pathway and the p53 tumour suppressor pathway. The present structure provides a framework for the design of inhibitory therapeutic strategies against oncogenesis mediated by human papilloma virus. PMID:26789255

  9. Oxidative Stress Induced Ventricular Arrhythmia and Impairment of Cardiac Function in Nos1ap Deleted Mice.

    PubMed

    Sugiyama, Koji; Sasano, Tetsuo; Kurokawa, Junko; Takahashi, Kentaro; Okamura, Tadashi; Kato, Norihiro; Isobe, Mitsuaki; Furukawa, Tetsushi

    2016-05-25

    Genome-wide association study has identified that the genetic variations at NOS1AP (neuronal nitric oxide synthase-1 adaptor protein) were associated with QT interval and sudden cardiac death (SCD). However, the mechanism linking a genetic variant of NOS1AP and SCD is poorly understood. We used Nos1ap knockout mice (Nos1ap(-/-)) to determine the involvement of Nos1ap in SCD, paying special attention to oxidative stress.At baseline, a surface electrocardiogram (ECG) and ultrasound echocardiography (UCG) showed no difference between Nos1ap(-/-) and wild-type (WT) mice. Oxidative stress was induced by a single injection of doxorubicin (Dox, 25 mg/kg). After Dox injection, Nos1ap(-/-) showed significantly higher mortality than WT (93.3 versus 16.0% at day 14, P < 0.01). ECG showed significantly longer QTc in Nos1ap(-/-) than WT, and UCG revealed significant reduction of fractional shortening (%FS) only in Nos1ap(-/-) after Dox injection. Spontaneous ventricular tachyarrhythmias were documented by telemetry recording after Dox injection only in Nos1ap(-/-). Ex vivo optical mapping revealed that the action potential duration (APD)90 was prolonged at baseline in Nos1ap(-/-), and administration of Dox lengthened APD90 more in Nos1ap(-/-) than in WT. The expression of Bnp and the H2O2 level were higher in Nos1ap(-/-) after Dox injection. Nos1ap(-/-) showed a reduced amplitude of calcium transient in isolated cardiomyocytes after Dox injection. Administration of the antioxidant N-acetyl-L-cysteine significantly reduced mortality of Nos1ap(-/-) by Dox injection, accompanied by prevention of QT prolongation and a reduction in %FS.Although Nos1ap(-/-) mice have apparently normal hearts, oxidative stress evokes ventricular tachyarrhythmia and heart failure, which may cause sudden cardiac death. PMID:27170476

  10. The driving mechanism of roAp stars : effects of global metallicity

    NASA Astrophysics Data System (ADS)

    Theado, S.; Dupret, M.-A.; Noels, A.

    2008-12-01

    We have investigated the influence of global metallicity on the excitation mechanism of roAp star pulsations. Our computations show that the opacity in the driving region of the roAp modes is strongly sensitive to the metal content but surprisingly the roAp theoretical instability strip is only weakly affected by metallicity changes.

  11. The driving mechanism of roAp stars : effects of local metallicity enhancement

    NASA Astrophysics Data System (ADS)

    Théado, S.; Dupret, M.-A.; Noels, A.

    2009-07-01

    We have investigated the influence of a local metallicity enhancement on the excitation mechanism of roAp star pulsations. Our computations show that such accumulations poorly affect the position of the theoretical roAp star instability strip although the opacity in the driving region of roAp modes is affected by metal accumulation.

  12. The U.S. Government's Assistance to the AP's World-Wide Expansion: 1912-1948.

    ERIC Educational Resources Information Center

    Renaud-Komiya, Jean-Luc

    A study of the extent of the diplomatic and commercial assistance provided by the United States government to the Associated Press (AP) from 1912 to 1948 shows AP's manager, Kent Cooper, to be less a champion of the free press than an efficient captain of industry in expanding AP influence across the globe. Early in the twentieth century, British,…

  13. Building Reading, Writing and Analysis in the AP U.S. History Classroom

    ERIC Educational Resources Information Center

    Heller, Stephen; Stacy, Jason

    2013-01-01

    The building of historical thinking skills has historically been a lonely endeavor for AP U.S. history teachers. Many often generate their own pedagogy, perhaps modified from an AP workshop or generally gleaned from released exam essay questions. However, as currently scheduled, in 2014, the AP U.S. history exam will undergo a redesign that will…

  14. The Drosophila melanogaster Mutants apblot and apXasta Affect an Essential apterous Wing Enhancer.

    PubMed

    Bieli, Dimitri; Kanca, Oguz; Gohl, Daryl; Denes, Alexandru; Schedl, Paul; Affolter, Markus; Müller, Martin

    2015-06-01

    The selector gene apterous (ap) plays a key role during the development of the Drosophila melanogaster wing because it governs the establishment of the dorsal-ventral (D-V) compartment boundary. The D-V compartment boundary is known to serve as an important signaling center that is essential for the growth of the wing. The role of Ap and its downstream effectors have been studied extensively. However, very little is known about the transcriptional regulation of ap during wing disc development. In this study, we present a first characterization of an essential wing-specific ap enhancer. First, we defined an 874-bp fragment about 10 kb upstream of the ap transcription start that faithfully recapitulates the expression pattern of ap in the wing imaginal disc. Analysis of deletions in the ap locus covering this element demonstrated that it is essential for proper regulation of ap and formation of the wing. Moreover, we showed that the mutations ap(blot) and ap(Xasta) directly affect the integrity of this enhancer, leading to characteristic wing phenotypes. Furthermore, we engineered an in situ rescue system at the endogenous ap gene locus, allowing us to investigate the role of enhancer fragments in their native environment. Using this system, we were able to demonstrate that the essential wing enhancer alone is not sufficient for normal wing development. The in situ rescue system will allow us to characterize the ap regulatory sequences in great detail at the endogenous locus. PMID:25840432

  15. 78 FR 21817 - Amendment of Restricted Area R-6601; Fort A.P. Hill, VA

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-12

    ... limits and increase the time of designation of restricted area R-6601, Fort A.P. Hill, VA, (77 FR 35308... Administration 14 CFR Part 73 RIN 2120-AA66 Amendment of Restricted Area R-6601; Fort A.P. Hill, VA AGENCY... limits and time of designation of restricted area R-6601, Fort A.P. Hill, VA. The U.S. Army...

  16. Using a Classroom Response System to Improve Multiple-Choice Performance in AP[R] Physics

    ERIC Educational Resources Information Center

    Bertrand, Peggy

    2009-01-01

    Participation in rigorous high school courses such as Advanced Placement (AP[R]) Physics increases the likelihood of college success, especially for students who are traditionally underserved. Tackling difficult multiple-choice exams should be part of any AP program because well-constructed multiple-choice questions, such as those on AP exams and…

  17. Insights into Association of the NuRD Complex with FOG-1 from the Crystal Structure of an RbAp48·FOG-1 Complex*

    PubMed Central

    Lejon, Sara; Thong, Sock Yue; Murthy, Andal; AlQarni, Saad; Murzina, Natalia V.; Blobel, Gerd A.; Laue, Ernest D.; Mackay, Joel P.

    2011-01-01

    Chromatin-modifying complexes such as the NuRD complex are recruited to particular genomic sites by gene-specific nuclear factors. Overall, however, little is known about the molecular basis for these interactions. Here, we present the 1.9 Å resolution crystal structure of the NuRD subunit RbAp48 bound to the 15 N-terminal amino acids of the GATA-1 cofactor FOG-1. The FOG-1 peptide contacts a negatively charged binding pocket on top of the RbAp48 β-propeller that is distinct from the binding surface used by RpAp48 to contact histone H4. We further show that RbAp48 interacts with the NuRD subunit MTA-1 via a surface that is distinct from its FOG-binding pocket, providing a first glimpse into the way in which NuRD assembly facilitates interactions with cofactors. Our RbAp48·FOG-1 structure provides insight into the molecular determinants of FOG-1-dependent association with the NuRD complex and into the links between transcription regulation and nucleosome remodeling. PMID:21047798

  18. AP-1 activity during normal human keratinocyte differentiation: evidence for a cytosolic modulator of AP-1/DNA binding.

    PubMed

    Briata, P; D'Anna, F; Franzi, A T; Gherzi, R

    1993-01-01

    Increased levels of c-fos and c-jun expression have been observed in differentiating epithelial cells. However, no data are available on activator protein 1 (AP-1) activity during keratinocyte differentiation. In this work we investigated c-fos and c-jun gene expression and AP-1-(12-O-tetradecanoylphorbol-13-acetate)-responsive enhancer element (TRE) binding activity during keratinocyte differentiation utilizing both authentic and in culture-reconstituted human epidermis. We demonstrate that: (i) in reconstituted epidermis, non-differentiated and differentiated keratinocytes express equivalent levels of c-Jun, while in reconstituted epidermis permanently grafted onto athymic mice, as well as in authentic epidermis, c-Jun is predominantly expressed in the granular layer of the tissue. Equivalent levels of c-fos expression have been found in all the layers of both reconstituted and authentic epidermis. (ii) Nuclear extracts from cultures enriched in differentiated keratinocytes display an 80-90% reduction of AP-1 activity when compared to extracts from cultures enriched in nondifferentiated cells. (iii) Cytosolic extracts obtained from cultures enriched in differentiated cells reduce, in a concentration-dependent manner, the AP-1 activity present in nuclear extracts of both mammalian and Drosophila cells. (iv) The specific TRE binding activity of a recombinant c-Jun protein is significantly reduced by cytosolic extracts of differentiated keratinocytes, while the specific DNA binding of the purified recombinant human homeoprotein HOX4B is not. (v) The dephosphorylation, by alkaline phosphatase, of cytosolic extracts increases the inhibitory activity already present or makes evident a latent activity. PMID:8416791

  19. PsAP2 an AP2/ERF family transcription factor from Papaver somniferum enhances abiotic and biotic stress tolerance in transgenic tobacco.

    PubMed

    Mishra, Sonal; Phukan, Ujjal J; Tripathi, Vineeta; Singh, Dhananjay K; Luqman, Suaib; Shukla, Rakesh Kumar

    2015-09-01

    The AP2/ERFs are one of the most important family of transcription factors which regulate multiple responses like stress, metabolism and development in plants. We isolated PsAP2 a novel AP2/ERF from Papaver somniferum which was highly upregulated in response to wounding followed by ethylene, methyl jasmonate and ABA treatment. PsAP2 showed specific binding with both DRE and GCC box elements and it was able to transactivate the reporter genes in yeast. PsAP2 overexpressing transgenic tobacco plants exhibited enhanced tolerance towards both abiotic and biotic stresses . Real time transcript expression analysis showed constitutive upregulation of tobacco Alternative oxidase1a and Myo-inositol-1-phosphate synthase in PsAP2 overexpressing tobacco plants. Further, PsAP2 showed interaction with NtAOX1a promoter in vitro, it also specifically activated the NtAOX1a promoter in yeast and tobacco BY2 cells. The silencing of PsAP2 using VIGS lead to significant reduction in the AOX1 level in P. somniferum. Taken together PsAP2 can directly bind and transcriptionally activate NtAOX1a and its overexpression in tobacco imparted increased tolerance towards both abiotic and biotic stress. PMID:26319514

  20. NRC confirmatory AP600 safety system phase I testing in the ROSA/AP600 test facility

    SciTech Connect

    Rhee, G.S.; Kukita, Yutaka; Schultz, R.R.

    1996-03-01

    The NRC confirmatory phase I testing for the AP600 safety systems has been completed in the modified ROSA (Rig of Safety Assessment) test facility located at the Japan Atomic Energy Research Institute (JAERI) campus in Tokai, Japan. The test matrix included a variety of accident scenarios covering both design and beyond-design basis accidents. The test results indicate the AP600 safety systems as reflected in ROSA appear to perform as designed and there is no danger of core heatup for the accident scenarios investigated. In addition, no detrimental system interactions nor adverse effects of non-safety systems on the safety system functions were identified. However, three phenomena of interest have been identified for further examination to determine whether they are relevant to the AP600 plant. Those three phenomena are: (1) a potential for water hammer caused by rapid condensation which may occur following the actuation of the automatic depressurization system (ADS), (2) a large thermal gradient in the cold leg pipe where cooled water returns from the passive residual heat removal system and forms a thermally stratified layer, and (3) system-wide oscillations initiating following the ADS stage 4 actuation and persisting until the liquid in the pressurizer drains and steam generation in the core becomes insignificant.

  1. AP Physics 1 & 2; Some Things Old and Some Things New

    NASA Astrophysics Data System (ADS)

    Morse, Robert

    2015-04-01

    In fall September 2014, AP Physics 1 and AP Physics 2 replaced the AP Physics B curriculum, with the first exams in the new courses coming up in May 2015. In this talk I will give an overview of the history and rationale for the changes, describe the process of developing the new Curriculum Framework, discuss changes in emphasis compared to AP Physics B, including the emphasis on laboratory work. Finally, I will give examples of the difference in the style of the questions to be used in the new AP exams.

  2. Packaging design criteria, transfer and disposal of 102-AP mixer pump

    SciTech Connect

    Carlstrom, R.F.

    1994-11-23

    A mixer pump installed in storage tank 241-AP-102 (102-AP) has failed. This pump is referred to as the 102-AP mixer pump (APMP). The APMP will be removed from 102-AP 1 and a new pump will be installed. The main purpose of the Packaging Design Criteria (PDC) is to establish criteria necessary to design and fabricate a shipping container for the transfer and storage of the APMP from 102-AP. The PDC will be used as a guide to develop a Safety Evaluation for Packaging (SEP).

  3. LDL immune complexes stimulate LDL receptor expression in U937 histiocytes via extracellular signal-regulated kinase and AP-1.

    PubMed

    Fu, Yuchang; Huang, Yan; Bandyopadhyay, Sumita; Virella, Gabriel; Lopes-Virella, Maria F

    2003-07-01

    We have previously shown that LDL-containing immune complexes (LDL-ICs) induce up-regulation of LDL receptor (LDLR) expression in human macrophages. The present study further investigated the molecular mechanisms leading to LDLR up-regulation by LDL-ICs as well as the signaling pathways involved. Results showed that treatment of U937 histiocytes with LDL-ICs did not increase the precursors and the cleaved forms of sterol-regulatory element binding proteins (SREBPs) 1a and 2, suggesting that SREBPs may not be involved in LDLR up-regulation by LDL-ICs. Promoter deletion and mutation studies showed that the AP-1 binding sites were essential for LDL-IC-stimulated LDLR expression. Electrophoretic mobility shift assays further demonstrated that LDL-ICs stimulated transcription factor AP-1 activity. Studies assessing the signaling pathways involved in LDLR up-regulation by LDL-ICs showed that the up-regulation of LDLR was extracellular signal-regulated kinase (ERK) dependent. In conclusion, the present study shows that LDL-ICs up-regulate LDLR expression via the ERK signaling pathway and the AP-1 motif-dependent transcriptional activation. PMID:12730303

  4. Scorpion Venom Heat-Resistant Peptide Attenuates Glial Fibrillary Acidic Protein Expression via c-Jun/AP-1.

    PubMed

    Cao, Zhen; Wu, Xue-Fei; Peng, Yan; Zhang, Rui; Li, Na; Yang, Jin-Yi; Zhang, Shu-Qin; Zhang, Wan-Qin; Zhao, Jie; Li, Shao

    2015-11-01

    Scorpion venom has been used in the Orient to treat central nervous system diseases for many years, and the protein/peptide toxins in Buthus martensii Karsch (BmK) venom are believed to be the effective components. Scorpion venom heat-resistant peptide (SVHRP) is an active component of the scorpion venom extracted from BmK. In a previous study, we found that SVHRP could inhibit the formation of a glial scar, which is characterized by enhanced glial fibrillary acidic protein (GFAP) expression, in the epileptic hippocampus. However, the cellular and molecular mechanisms underlying this process remain to be clarified. The results of the present study indicate that endogenous GFAP expression in primary rat astrocytes was attenuated by SVHRP. We further demonstrate that the suppression of GFAP was primarily mediated by inhibiting both c-Jun expression and its binding with AP-1 DNA binding site and other factors at the GFAP promoter. These results support that SVHRP contributes to reducing GFAP at least in part by decreasing the activity of the transcription factor AP-1. In conclusion, the effects of SVHRP on astrocytes with respect to the c-Jun/AP-1 signaling pathway in vitro provide a practical basis for studying astrocyte activation and inhibition and a scientific basis for further studies of traditional medicine. PMID:26134308

  5. NF-κB/AP-1-targeted inhibition of macrophage-mediated inflammatory responses by depigmenting compound AP736 derived from natural 1,3-diphenylpropane skeleton.

    PubMed

    Ha, Van Thai; Beak, Heung Soo; Kim, Eunji; Baek, Kwang-Soo; Hossen, Muhammad Jahangir; Yang, Woo Seok; Kim, Yong; Kim, Jun Ho; Yang, Sungjae; Kim, Jeong-Hwan; Joo, Yung Hyup; Lee, Chang Seok; Choi, Joonho; Shin, Hong-Ju; Hong, Sungyoul; Shin, Song Seok; Cho, Jae Youl

    2014-01-01

    AP736 was identified as an antimelanogenic drug that can be used for the prevention of melasma, freckles, and dark spots in skin by acting as a suppressor of melanin synthesis and tyrosinase expression. Since macrophage-mediated inflammatory responses are critical for skin health, here we investigated the potential anti-inflammatory activity of AP736. The effects of AP736 on various inflammatory events such as nitric oxide (NO)/prostaglandin (PG) E2 production, inflammatory gene expression, phagocytic uptake, and morphological changes were examined in RAW264.7 cells. AP736 was found to strongly inhibit the production of both NO and PGE2 in lipopolysaccharide- (LPS-) treated RAW264.7 cells. In addition, AP736 strongly inhibited both LPS-induced morphological changes and FITC-dextran-induced phagocytic uptake. Furthermore, AP736 also downregulated the expression of multiple inflammatory genes, such as inducible NO synthase (iNOS), cyclooxygenase- (COX-) 2, and interleukin- (IL-) 1β in LPS-treated RAW264.7 cells. Transcription factor analysis, including upstream signalling events, revealed that both NF-κB and AP-1 were targeted by AP736 via inhibition of the IKK/IκBα and IRAK1/TAK1 pathways. Therefore, our results strongly suggest that AP736 is a potential anti-inflammatory drug due to its suppression of NF-κB-IKK/IκBα and AP-1-IRAK1/TAK1 signalling, which may make AP736 useful for the treatment of macrophage-mediated skin inflammation. PMID:25386046

  6. The Drosophila melanogaster Mutants apblot and apXasta Affect an Essential apterous Wing Enhancer

    PubMed Central

    Bieli, Dimitri; Kanca, Oguz; Gohl, Daryl; Denes, Alexandru; Schedl, Paul; Affolter, Markus; Müller, Martin

    2015-01-01

    The selector gene apterous (ap) plays a key role during the development of the Drosophila melanogaster wing because it governs the establishment of the dorsal-ventral (D-V) compartment boundary. The D-V compartment boundary is known to serve as an important signaling center that is essential for the growth of the wing. The role of Ap and its downstream effectors have been studied extensively. However, very little is known about the transcriptional regulation of ap during wing disc development. In this study, we present a first characterization of an essential wing-specific ap enhancer. First, we defined an 874-bp fragment about 10 kb upstream of the ap transcription start that faithfully recapitulates the expression pattern of ap in the wing imaginal disc. Analysis of deletions in the ap locus covering this element demonstrated that it is essential for proper regulation of ap and formation of the wing. Moreover, we showed that the mutations apblot and apXasta directly affect the integrity of this enhancer, leading to characteristic wing phenotypes. Furthermore, we engineered an in situ rescue system at the endogenous ap gene locus, allowing us to investigate the role of enhancer fragments in their native environment. Using this system, we were able to demonstrate that the essential wing enhancer alone is not sufficient for normal wing development. The in situ rescue system will allow us to characterize the ap regulatory sequences in great detail at the endogenous locus. PMID:25840432

  7. Molecular Basis for Enhancement of the Meiotic DMCI Recombinase by RAD51AP1

    SciTech Connect

    Dray, Eloise; Dunlop, Myun Hwa; Kauppi, Liisa; San Filippo, Joseph San; Wiese, Claudia; Tsai, Miaw-Sheue; Begovic, Sead; Schild, David; Jasin, Maria; Keeney, Scott; Sung, Patrick

    2010-11-05

    Homologous recombination is needed for meiotic chromosome segregation, genome maintenance, and tumor suppression. RAD51AP1 (RAD51 Associated Protein 1) has been shown to interact with and enhance the recombinase activity of RAD51. Accordingly, genetic ablation of RAD51AP1 leads to enhanced sensitivity to and also chromosome aberrations upon DNA damage, demonstrating a role for RAD51AP1 in mitotic homologous recombination. Here we show physical association of RAD51AP1 with the meiosis-specific recombinase DMC1 and a stimulatory effect of RAD51AP1 on the DMC1-mediated D-loop reaction. Mechanistic studies have revealed that RAD51AP1 enhances the ability of the DMC1 presynaptic filament to capture the duplex DNA partner and to assemble the synaptic complex, in which the recombining DNA strands are homologously aligned. We also provide evidence that functional co-operation is dependent on complex formation between DMC1 and RAD51AP1, and that distinct epitopes in RAD51AP1 mediate interactions with RAD51 and DMC1. Finally, we show that RAD51AP1 is expressed in mouse testes, and that RAD51AP1 foci co-localize with a subset of DMC1 foci in spermatocytes. These results suggest that RAD51AP1 also serves an important role in meiotic homologous recombination.

  8. AP4 is a mediator of epithelial–mesenchymal transition and metastasis in colorectal cancer

    PubMed Central

    Jackstadt, Rene; Röh, Simone; Neumann, Jens; Jung, Peter; Hoffmann, Reinhard; Horst, David; Berens, Christian; Bornkamm, Georg W.; Kirchner, Thomas; Menssen, Antje

    2013-01-01

    The basic helix-loop-helix transcription factor AP4/TFAP4/AP-4 is encoded by a c-MYC target gene and displays up-regulation concomitantly with c-MYC in colorectal cancer (CRC) and numerous other tumor types. Here a genome-wide characterization of AP4 DNA binding and mRNA expression was performed using a combination of microarray, genome-wide chromatin immunoprecipitation, next-generation sequencing, and bioinformatic analyses. Thereby, hundreds of induced and repressed AP4 target genes were identified. Besides many genes involved in the control of proliferation, the AP4 target genes included markers of stemness (LGR5 and CD44) and epithelial–mesenchymal transition (EMT) such as SNAIL, E-cadherin/CDH1, OCLN, VIM, FN1, and the Claudins 1, 4, and 7. Accordingly, activation of AP4 induced EMT and enhanced migration and invasion of CRC cells. Conversely, down-regulation of AP4 resulted in mesenchymal–epithelial transition and inhibited migration and invasion. In addition, AP4 induction was required for EMT, migration, and invasion caused by ectopic expression of c-MYC. Inhibition of AP4 in CRC cells resulted in decreased lung metastasis in mice. Elevated AP4 expression in primary CRC significantly correlated with liver metastasis and poor patient survival. These findings imply AP4 as a new regulator of EMT that contributes to metastatic processes in CRC and presumably other carcinomas. PMID:23752226

  9. Multiplicity among chemically peculiar stars. II. Cool magnetic Ap stars

    NASA Astrophysics Data System (ADS)

    Carrier, F.; North, P.; Udry, S.; Babel, J.

    2002-10-01

    We present new orbits for sixteen Ap spectroscopic binaries, four of which might in fact be Am stars, and give their orbital elements. Four of them are SB2 systems: HD 5550, HD 22128, HD 56495 and HD 98088. The twelve other stars are: HD 9996, HD 12288, HD 40711, HD 54908, HD 65339, HD 73709, HD 105680, HD 138426, HD 184471, HD 188854, HD 200405 and HD 216533. Rough estimates of the individual masses of the components of HD 65339 (53 Cam) are given, combining our radial velocities with the results of speckle interferometry and with Hipparcos parallaxes. Considering the mass functions of 74 spectroscopic binaries from this work and from the literature, we conclude that the distribution of the mass ratio is the same for cool Ap stars and for normal G dwarfs. Therefore, the only differences between binaries with normal stars and those hosting an Ap star lie in the period distribution: except for the case of HD 200405, all orbital periods are longer than (or equal to) 3 days. A consequence of this peculiar distribution is a deficit of null eccentricities. There is no indication that the secondary has a special nature, like e.g. a white dwarf. Based on observations collected at the Observatoire de Haute-Provence (CNRS), France. Tables 1 to 3 are only available in electronic form at the CDS via anonymous ftp to cdsarc.u-strasbg.fr (130.79.128.5) or via http://cdsweb.u-strasbg.fr/cgi-bin/qcat?J/A+A/394/151 Appendix B is only available in electronic form at http://www.edpsciences.org

  10. Involvement of apurinic sites in the synergistic action of alkylating and intercalating drugs in Escherichia coli.

    PubMed

    Malvy, C; Safraoui, H; Bloch, E; Bertrand, J R

    1988-03-01

    The toxicity of the intercalating compounds 9-aminoellipticine (9AE) and isopropyl-oxazolopyridocarbazole (Ipr-OPC) were studied. The inhibitory effect of non-toxic doses of 9AE, which incises DNA at apurinic (AP) sites, or Ipr-OPC, which does not cleave DNA at AP sites, with non-toxic doses of the alkylating agent dimethylsulphate (DMS) on the growth of Escherichia coli strain AB1157, is additive. The same result has been observed with an exonuclease III mutant which has only 10% of the AP endonuclease activity. However, 9AE or Ipr-OPC display a synergistic toxic effect with a DMS concentration which allows 20% of E. coli AB1157 survival. This synergy is increased for 9AE in the AP endonuclease mutant when compared to the wild-type strain. Under identical conditions 9AE and Ipr-OPC have no synergistic effect on a mutant deficient in the enzymes which generate AP sites. Therefore AP sites are involved in the synergistic toxicity of DMS and the studied intercalating agents. However, the precise role of the interaction of intercalating agents with AP sites, either without cleavage (type 1 compounds) or with cleavage (type 2 compounds), in the observed effect remains an open question. PMID:3284541

  11. APS beamline standard components handbook, Version 1. 3

    SciTech Connect

    Hahn, U.; Shu, D.; Kuzay, T.M.

    1993-02-01

    This Handbook in its current version (1.3) contains descriptions, specifications, and preliminary engineering design drawings for many of the standard components. The design status and schedules have been provided wherever possible. In the near future, the APS plans to update engineering drawings of identified standard beamline components and complete the Handbook. The completed version of this Handbook will become available to both the CATs and potential vendors. Use of standard components should result in major cost reductions for CATs in the areas of beamline design and construction.

  12. Abundance analysis of roAp stars. II. HD 203932

    NASA Astrophysics Data System (ADS)

    Gelbmann, M.; Kupka, F.; Weiss, W. W.; Mathys, G.

    1997-03-01

    A new tool to simplify abundance analyses which is based on stand-alone programs has been applied to the rapidly oscillating Ap star HD 203932 (BI Mic, CD -30 18600, SAO 212996; Ap(SrEu), V=8.82mag). The spectroscopically determined T_eff_=7450+/-100K and logg=4.3+/-0.15 put this star close to the ZAMS. Other fundamental atmospheric parameters are v_micro_<0.6km/s and the total abundance of all iron peak elements [M/H]=0.0+/-0.1. The fundamental parameters put HD 203932 in a region of the HR-diagram where convection starts becoming efficient and the standard mixing length theory models lead to severe problems in the determination of the atmospheric parameters. The difference between the upper limit for logg obtained from several variants of the mixing length theory and the Canuto-Mazzitelli model indicates that the choice of a particular convection model can influence the determination of basic stellar parameters. For the first time abundances were determined for HD 203932 showing a pattern for the 35 investigated elements which is similar to α Cir (Kupka et al. 1996A&A...308..886K, Paper I). Fe and Ni have about solar abundance, Cr and especially Co are clearly overabundant as well as rare earth elements. The most underabundant element is Sc, followed by C, N, and O, which is a common property of CP2 stars. The lack of a correlation in our data between individual line abundances and their effective Lande factors implies a mean magnetic field modulus not exceeding few kG. Compared to the last homogeneous spectroscopic investigation of a large sample of chemically peculiar stars (21 cool Ap stars, Adelman 1973ApJ...183...95A), our analysis is based on data with higher spectral resolution and signal-to-noise ratio. Even more important, we are using a much larger atomic line data base with more precise atomic parameters than available more than twenty years ago.

  13. Euphorbesulins A-P, Structurally Diverse Diterpenoids from Euphorbia esula.

    PubMed

    Zhou, Bin; Wu, Yan; Dalal, Seema; Cassera, Maria B; Yue, Jian-Min

    2016-08-26

    Aqueous ethanol extracts of powdered twigs of Euphorbia esula afforded 16 new diterpenoids, named euphorbesulins A-P. These euphorbesulins included presegetane (1-3), jatrophane (4-14), paraliane (15), and isopimarane (16) diterpenoids as well as six known analogues. Compounds 1-3 represent a rare type of presegetane diterpenoid. Their structures were determined by analysis of the spectroscopic data, and the absolute configuration of 1 was established by X-ray crystallography. Diterpenoid 7 showed low nanomolar antimalarial activity, while the remaining compounds showed only moderate or no antimalarial activity. PMID:27447736

  14. The driving mechanism of roAp stars

    NASA Astrophysics Data System (ADS)

    Dupret, M.-A.; Théado, S.; Noels, A.

    2008-10-01

    We analyse in detail the driving mechanism of roAp stars and present the theoretical instability strip predicted by our models with solar metallicity. A particular attention is given to the interpretation of the role played by the different eigenfunctions in the stabilization of the modes at the red edge of the instability strip. The gradient of temperature in the HI opacity bump appears to play a major role in this context. We also consider the particular and complex role played by the shape of the eigenfunctions (location of the nodes, ...).

  15. The APS transfer line from linac to injector synchrotron

    SciTech Connect

    Koul, R.K.; Crosbie, E.

    1991-03-01

    This note describes the low-energy-transfer-line designed for the APS. The low energy transfer line constitutes two transport lines. One of these lines runs from linac to the positron accumulator ring, also called ``PAR``, and is 23.7138 m long. The second part of the low energy transport line runs from the ``PAR`` to the injector synchrtoron and is about 30.919 m long. The above length includes two quadrupoles, a bend magnet and a septum magnet in the injector synchrotron.

  16. Long period oscillations in roAp stars

    NASA Astrophysics Data System (ADS)

    Riley, J. D.; Kurtz, D. W.; Cunha, M. S.

    2004-12-01

    We present the results of observations made over three weeks using the UCT CCD Photometer on the 0.75-m telescope at the South African Astronomical Observatory. Candidate long period roAp stars were identified from their positions on the H-R diagram and observed for a typical period of 4 hr to test for the existence of pulsations, with particular emphasis on pulsations with periods in excess of 15 min. Although 13 stars were successfully observed, none exhibited significant pulsations.

  17. Effects of AP particle size on combustion response to crossflow

    NASA Technical Reports Server (NTRS)

    Cohen, N. S.; Strand, L. D.

    1983-01-01

    An analytical model is developed for the linearized velocity-coupled combustion response function. The model treats elements of response to perturbations in pressure, in composition due to the heterogeneity of composite propellants, and in crossflow velocity. The effects of AP particle size are accounted for in terms of effects on controlling ballistics properties and in terms of fluctuations in propellant composition. There are two facets of the crossflow problem: the effect of crossflow velocity on the various response elements, and the response to velocity perturbations. Both are dealt with in this paper. Important trends derived from series of parametric computations are described.

  18. Operation of the APS photoinjector drive laser system.

    SciTech Connect

    Li, Y.; Accelerator Systems Division

    2008-08-04

    The APS photoinjector drive laser system has been in operation since 1999 and is achieving a performance level exceeding the requirement of stable operation of the LEUTL FEL system. One remarkable number is the UV energy stability of better than 2% rms, sometimes less than 1% rms. This report summarizes the operation experience of the laser system and the improvements made along the way. We also outline the route of upgrade of the system and some frontier laser research and development opportunities in ultrabright electron beam generation.

  19. Upgraded cavities for the positron accumulator ring of the APS

    SciTech Connect

    Kang, Y.W.; Jiang, X.; Mangra, D.

    1997-08-01

    Upgraded versions of cavities for the APS positron accumulator ring (PAR) have been built and are being tested. Two cavities are in the PAR: a fundamental 9.8-MHz cavity and a twelfth harmonic 117.3-MHz cavity. Both cavities have been manufactured for higher voltage operation with improved Q-factors, reliability, and tuning capability. Both cavities employ current-controlled ferrite tuners for control of the resonant frequency. The harmonic cavity can be operated in either a pulsed mode or a CW mode. The rf properties of the cavities are presented.

  20. H-beta line variability in magnetic Ap stars. I

    NASA Technical Reports Server (NTRS)

    Madej, J.; Jahn, K.; Stepien, K.

    1984-01-01

    Preliminary results of photometric measurements of H-beta in several Ap stars are presented. Periodic variations are found certainly in Theta Aur and Alpha (2) CVn, and possibly in Phi Dra. For the other stars upper limits for variations of H-beta are determined. Observed amplitudes are transformed into variations of equivalent width assuming specific profile variations. The results show that variations of equivalent width of H-beta in the stars investigated are of the order of 10 percent or less.

  1. Initial diagnostics commissioning results for the Advanced Photon Source (APS)

    SciTech Connect

    Lumpkin, A.; Patterson, D.; Wang, X.

    1995-07-01

    Principal diagnostics systems have been installed and nearly all have been commissioned on the subsystems of the Advanced Photon Source (APS) facility. Data have been obtained on beam position, beam profile, current, beam loss rate, and synchrotron radiation monitors on both injector rings and most recently the main 7-GeV storage ring. Results for the 150- to 450-MeV electron beams in the accumulator ring, up to 7 GeV in the injector synchrotron, and 4.5 to 7 GeV in the SR will be presented.

  2. S100P/RAGE signaling regulates microRNA-155 expression via AP-1 activation in colon cancer

    PubMed Central

    Onyeagucha, Benjamin Chidi; Mercado-Pimentel, Melania E.; Hutchison, Jennifer; Flemington, Erik K.; Nelson, Mark A.

    2013-01-01

    Accumulating evidence indicates that elevated S100P promotes the pathogenesis of cancers, including colon cancer. S100P exerts its effects by binding to and activating the Receptor for Advance Glycation End-products (RAGE). The effects of up-regulated S100P/RAGE signaling on cell functions are well documented. Despite these observations, little is known about the downstream targets of S100P/RAGE signaling. In the present study, we demonstrated for the first time that activation of RAGE by S100P regulates oncogenic microRNA-155 (miR-155) expression through Activator Protein-1 (AP-1) stimulation in colon cancer cells. Ectopic S100P up-regulated miR-155 levels in human colon cancer cells. Conversely, knockdown of S100P resulted in a decrease in miR-155 levels. Exogenous S100P induced miR-155 expression, but blockage of the RAGE with anti-RAGE antibody suppressed the induction of miR-155 by exogenous S100P. Attenuation of AP-1 activation through pharmacological inhibition of MEK activation or genetic inhibition of c-Jun activation using dominant negative c-Jun (TAM67) suppressed miR-155 induction by exogenous S100P. Also, S100P treatment stimulated the enrichment of c-Fos, an AP-1 family member, at the miR-155 host gene promoter site. Finally, a functional study demonstrated that miR-155 knockdown decreases colon cancer cell growth, motility, and invasion. Altogether, these data demonstrate that the expression of miR-155 is regulated by S100P and is dependent on RAGE activation and stimulation of AP-1. PMID:23693020

  3. X Linkage of AP3A, a Homolog of the Y-Linked MADS-Box Gene AP3Y in Silene latifolia and S. dioica

    PubMed Central

    Penny, Rebecca H.; Montgomery, Benjamin R.; Delph, Lynda F.

    2011-01-01

    Background The duplication of autosomal genes onto the Y chromosome may be an important element in the evolution of sexual dimorphism.A previous cytological study reported on a putative example of such a duplication event in a dioecious tribe of Silene (Caryophyllaceae): it was inferred that the Y-linked MADS-box gene AP3Y originated from a duplication of the reportedly autosomal orthologAP3A. However, a recent study, also using cytological methods, indicated that AP3A is X-linked in Silenelatifolia. Methodology/Principal Findings In this study, we hybridized S. latifolia and S. dioicato investigate whether the pattern of X linkage is consistent among distinct populations, occurs in both species, and is robust to genetic methods. We found inheritance patterns indicative of X linkage of AP3A in widely distributed populations of both species. Conclusions/Significance X linkage ofAP3A and Y linkage of AP3Yin both species indicates that the genes' ancestral progenitor resided on the autosomes that gave rise to the sex chromosomesand that neither gene has moved between chromosomes since species divergence.Consequently, our results do not support the contention that inter-chromosomal gene transfer occurred in the evolution of SlAP3Y from SlAP3A. PMID:21533056

  4. Draft Genome Sequences of Clinical Daptomycin-Nonsusceptible Methicillin-Resistant Staphylococcus aureus Strain APS211 and Its Daptomycin-Susceptible Progenitor APS210.

    PubMed

    Cameron, David R; Jiang, Jhih-Hang; Abbott, Iain J; Spelman, Denis W; Peleg, Anton Y

    2015-01-01

    To assess the genetic factors contributing to daptomycin resistance in Staphylococcus aureus, the draft genome of a clinically derived daptomycin-nonsusceptible isolate APS211 was generated and compared to the draft sequence of its susceptible progenitor strain APS210. Four genetic differences were identified including a previously described mutation within the mprF gene. PMID:26067951

  5. Draft Genome Sequences of Clinical Daptomycin-Nonsusceptible Methicillin-Resistant Staphylococcus aureus (MRSA) Strain APS211 and Its Daptomycin-Susceptible Progenitor APS210

    PubMed Central

    Cameron, David R.; Jiang, Jhih-Hang; Abbott, Iain J.; Spelman, Denis W.

    2015-01-01

    To assess the genetic factors contributing to daptomycin resistance in Staphylococcus aureus, the draft genome of a clinically derived daptomycin-nonsusceptible isolate APS211 was generated and compared to the draft sequence of its susceptible progenitor strain APS210. Four genetic differences were identified including a previously described mutation within the mprF gene. PMID:26067951

  6. Implementation and Initial Validation of the APS English Test [and] The APS English-Writing Test at Golden West College: Evidence for Predictive Validity.

    ERIC Educational Resources Information Center

    Isonio, Steven

    In May 1991, Golden West College (California) conducted a validation study of the English portion of the Assessment and Placement Services for Community Colleges (APS), followed by a predictive validity study in July 1991. The initial study was designed to aid in the implementation of the new test at GWC by comparing data on APS use at other…

  7. Incorporating Ninth-Grade PSAT/NMSQT® Scores into AP Potential™ Predictions for AP® European History and AP World History. Statistical Report 2014-1

    ERIC Educational Resources Information Center

    Zhang, Xiuyuan; Patel, Priyank; Ewing, Maureen

    2015-01-01

    Historically, AP Potential™ correlations and expectancy tables have been based on 10th-and 11th-grade PSAT/NMSQT® examinees and 11th-and 12th-grade AP® examinees for all subjects (Zhang, Patel, & Ewing,2014; Ewing, Camara, & Millsap, 2006; Camara & Millsap, 1998). However, a large number of students take AP European History and AP…

  8. Two peptides, TsAP-1 and TsAP-2, from the venom of the Brazilian yellow scorpion, Tityus serrulatus: evaluation of their antimicrobial and anticancer activities.

    PubMed

    Guo, Xiaoxiao; Ma, Chengbang; Du, Qiang; Wei, Ran; Wang, Lei; Zhou, Mei; Chen, Tianbao; Shaw, Chris

    2013-09-01

    Here we report two novel 17-mer amidated linear peptides (TsAP-1 and TsAP-2) whose structures were deduced from cDNAs cloned from a venom-derived cDNA library of the Brazilian yellow scorpion, Tityus serrulatus. Both mature peptides were structurally-characterised following their location in chromatographic fractions of venom and synthetic replicates of each were subjected to a range of biological assays. The peptides were each active against model test micro-organisms but with different potencies. TsAP-1 was of low potency against all three test organisms (MICs 120-160 μM), whereas TsAP-2 was of high potency against the Gram-positive bacterium, Staphylococcus aureus (MIC 5 μM) and the yeast, Candida albicans (10 μM). Haemolytic activity of TsAP-1 was low (4% at 160 μM) and in contrast, that of TsAP-2 was considerably higher (18% at 20 μM). Substitution of four neutral amino acid residues with Lys residues in each peptide had dramatic effects on their antimicrobial potencies and haemolytic activities, particularly those of TsAP-1. The MICs of the enhanced cationic analogue (TsAP-S1) were 2.5 μM for S. aureus/C. albicans and 5 μM for E. coli but with an associated large increase in haemolytic activity (30% at 5 μM). The same Lys residue substitutions in TsAP-2 produced a dramatic effect on its MIC for E. coli lowering this from >320 μM to 5 μM. TsAP-1 was ineffective against three of the five human cancer cell lines tested while TsAP-2 inhibited the growth of all five. Lys residue substitution of both peptides enhanced their potency against all five cell lines with TsAp-S2 being the most potent with IC50 values ranging between 0.83 and 2.0 μM. TsAP-1 and TsAP-2 are novel scorpion venom peptides with broad spectrum antimicrobial and anticancer cell activities the potencies of which can be significantly enhanced by increasing their cationicity. PMID:23770440

  9. RAD51AP2, a novel vertebrate- and meiotic-specific protein, sharesa conserved RAD51-interacting C-terminal domain with RAD51AP1/PIR51

    SciTech Connect

    Kovalenko, Oleg V.; Wiese, Claudia; Schild, David

    2006-07-25

    Many interacting proteins regulate and/or assist the activities of RAD51, a recombinase which plays a critical role in both DNA repair and meiotic recombination. Yeast two-hybrid screening of a human testis cDNA library revealed a new protein, RAD51AP2 (RAD51 Associated Protein 2), that interacts strongly with RAD51. A full-length cDNA clone predicts a novel vertebrate specific protein of 1159 residues, and the RAD51AP2 transcript was observed only in meiotic tissue (i.e. adult testis and fetal ovary), suggesting a meiotic-specific function for RAD51AP2. In HEK293 cells the interaction of RAD51 with an ectopically-expressed recombinant large fragment of RAD51AP2 requires the C-terminal 57 residues of RAD51AP2. This RAD51-binding region shows 81% homology to the C-terminus of RAD51AP1/PIR51, an otherwise totally unrelated RAD51-binding partner that is ubiquitously expressed. Analyses using truncations and point mutations in both RAD51AP1 and RAD51AP2 demonstrate that these proteins use the same structural motif for RAD51 binding. RAD54 shares some homology with this RAD51-binding motif, but this homologous region plays only an accessory role to the adjacent main RAD51-interacting region, which has been narrowed here to 40 amino acids. A novel protein, RAD51AP2, has been discovered that interacts with RAD51 through a C-terminal motif also present in RAD51AP1.

  10. Particle velocity measurements in HVOF and APS systems

    SciTech Connect

    Knight, R.; Smith, R.W.; Xiao, Z.; Hoffman, T.T.

    1994-12-31

    Production of reliable, repeatable coatings requires precise control of the process used to deposit them. Significant advances have recently been made in controlling the inputs to thermal spray processes, however, much work remains to be done to control process outputs and to correlate these with coatings characteristics. Thermal spray processes comprise the heating/melting, acceleration, impact, rapid solidification and incremental build-up of a large number of individual particles. Particle velocity is a key process parameter in determining coating properties such as density/porosity, bond strength and residual stress. Laser Stroboscopy and optical image analysis techniques have been used to image particles traveling in high velocity oxy-fuel (HVOF) and air plasma spray (APS) jets. Results indicate that these techniques can be used to measure particle velocity, trajectory and velocity distribution(s) in thermal spray jets. mean particle velocities of {approximately}400 m/s and {approximately}100 m/s have been measured for HVOF and APS respectively.

  11. The LOCA performance of the AP600 passive safety systems

    SciTech Connect

    Kemper, R.M.; Hochreiter, L.E.; Takeuchi, K.; Garner, D.C.; Nguyen, S.B.; Cunningham, J.P. ); Lee, S.N.K.; Tehrani, A.A.K.; Yang, H.; Bratby, P.A.W. )

    1992-01-01

    The AP600 is an advanced passive safeguards pressurized water reactor (PWR) that is being developed jointly by Westinghouse Electric Corporation, the U.S. Department of Energy, and the Electrical Power Research Institute. The plant has a thermal rating of 1940 MW (thermal) [600 MW(electric)] and has been designed with passive safeguard systems that utilize gravity feed injection rather than safety-grade active pumps and equipment. Calculations performed for a range of break sizes used locations to find the worst set of conditions for depressurizing the reactor coolant system. The main criterion was system inventory such that the core remained covered. The resulting break spectrum study indicates only that the double-ended guillotine shear of the direct vessel injection line (a .68-in. line that feeds the emergency core coolant flow into the vessel) resulted in a momentary core uncover. For all other small-break cases, the core remained covered as the reactor coolant system depressurized. The passive safety systems provided sufficient mass flow to the reactor vessel such that even under the more conservative Appendix K assumptions, the core remained covered and in a coolable state. The LOCA analysis performed for the AP600 confirms that passive safety systems can provide the core cooling necessary to meet the requirements of 10CFR50.46 with ample margin.

  12. Luteolin, a flavonoid, inhibits AP-1 activation by basophils

    SciTech Connect

    Hirano, Toru; Higa, Shinji; Arimitsu, Junsuke; Naka, Tetsuji; Ogata, Atsushi; Shima, Yoshihito; Fujimoto, Minoru; Yamadori, Tomoki; Ohkawara, Tomoharu; Kuwabara, Yusuke; Kawai, Mari; Matsuda, Hisashi; Yoshikawa, Masayuki; Maezaki, Naoyoshi; Tanaka, Tetsuaki; Kawase, Ichiro; Tanaka, Toshio . E-mail: ttanak@imed3.med.osaka-u.ac.jp

    2006-02-03

    Flavonoids including luteolin, apigenin, and fisetin are inhibitors of IL-4 synthesis and CD40 ligand expression by basophils. This study was done to search for compounds with greater inhibitory activity of IL-4 expression and to clarify the molecular mechanisms through which flavonoids inhibit their expression. Of the 37 flavonoids and related compounds examined, ayanin, luteolin, and apigenin were the strongest inhibitors of IL-4 production by purified basophils in response to anti-IgE antibody plus IL-3. Luteolin did not suppress Syk or Lyn phosphorylation in basophils, nor did suppress p54/46 SAPK/JNK, p38 MAPK, and p44/42 MAPK activation by a basophilic cell line, KU812 cells, stimulated with A23187 and PMA. However, luteolin did inhibit phosphorylation of c-Jun and DNA binding activity of AP-1 in nuclear lysates from stimulated KU812 cells. These results provide a fundamental structure of flavonoids for IL-4 inhibition and demonstrate a novel action of flavonoids that suppresses the activation of AP-1.

  13. Loss of JUNB/AP-1 promotes invasive prostate cancer

    PubMed Central

    Thomsen, M K; Bakiri, L; Hasenfuss, S C; Wu, H; Morente, M; Wagner, E F

    2015-01-01

    Prostate cancer is a frequent cause of male death in the Western world. Relatively few genetic alterations have been identified, likely owing to disease heterogeneity. Here, we show that the transcription factor JUNB/AP-1 limits prostate cancer progression. JUNB expression is increased in low-grade prostate cancer compared with normal human prostate, but downregulated in high-grade samples and further decreased in all metastatic samples. To model the hypothesis that this downregulation is functionally significant, we genetically inactivated Junb in the prostate epithelium of mice. When combined with Pten (phosphatase and tensin homologue) loss, double-mutant mice were prone to invasive cancer development. Importantly, invasive tumours also developed when Junb and Pten were inactivated in a small cell population of the adult anterior prostate by topical Cre recombinase delivery. The resulting tumours displayed strong histological similarity with human prostate cancer. Loss of JunB expression led to increased proliferation and decreased senescence, likely owing to decreased p16Ink4a and p21CIP1 in epithelial cells. Furthermore, the tumour stroma was altered with increased osteopontin and S100 calcium-binding protein A8/9 expression, which correlated with poor prognoses in patients. These data demonstrate that JUNB/AP-1 cooperates with PTEN signalling as barriers to invasive prostate cancer, whose concomitant genetic or epigenetic suppression induce malignant progression. PMID:25526087

  14. Loss of JUNB/AP-1 promotes invasive prostate cancer.

    PubMed

    Thomsen, M K; Bakiri, L; Hasenfuss, S C; Wu, H; Morente, M; Wagner, E F

    2015-04-01

    Prostate cancer is a frequent cause of male death in the Western world. Relatively few genetic alterations have been identified, likely owing to disease heterogeneity. Here, we show that the transcription factor JUNB/AP-1 limits prostate cancer progression. JUNB expression is increased in low-grade prostate cancer compared with normal human prostate, but downregulated in high-grade samples and further decreased in all metastatic samples. To model the hypothesis that this downregulation is functionally significant, we genetically inactivated Junb in the prostate epithelium of mice. When combined with Pten (phosphatase and tensin homologue) loss, double-mutant mice were prone to invasive cancer development. Importantly, invasive tumours also developed when Junb and Pten were inactivated in a small cell population of the adult anterior prostate by topical Cre recombinase delivery. The resulting tumours displayed strong histological similarity with human prostate cancer. Loss of JunB expression led to increased proliferation and decreased senescence, likely owing to decreased p16(Ink4a) and p21(CIP1) in epithelial cells. Furthermore, the tumour stroma was altered with increased osteopontin and S100 calcium-binding protein A8/9 expression, which correlated with poor prognoses in patients. These data demonstrate that JUNB/AP-1 cooperates with PTEN signalling as barriers to invasive prostate cancer, whose concomitant genetic or epigenetic suppression induce malignant progression. PMID:25526087

  15. The new AP Physics exams: Integrating qualitative and quantitative reasoning

    NASA Astrophysics Data System (ADS)

    Elby, Andrew

    2015-04-01

    When physics instructors and education researchers emphasize the importance of integrating qualitative and quantitative reasoning in problem solving, they usually mean using those types of reasoning serially and separately: first students should analyze the physical situation qualitatively/conceptually to figure out the relevant equations, then they should process those equations quantitatively to generate a solution, and finally they should use qualitative reasoning to check that answer for plausibility (Heller, Keith, & Anderson, 1992). The new AP Physics 1 and 2 exams will, of course, reward this approach to problem solving. But one kind of free response question will demand and reward a further integration of qualitative and quantitative reasoning, namely mathematical modeling and sense-making--inventing new equations to capture a physical situation and focusing on proportionalities, inverse proportionalities, and other functional relations to infer what the equation ``says'' about the physical world. In this talk, I discuss examples of these qualitative-quantitative translation questions, highlighting how they differ from both standard quantitative and standard qualitative questions. I then discuss the kinds of modeling activities that can help AP and college students develop these skills and habits of mind.

  16. Designing an artificial pancreas architecture: the AP@home experience.

    PubMed

    Lanzola, Giordano; Toffanin, Chiara; Di Palma, Federico; Del Favero, Simone; Magni, Lalo; Bellazzi, Riccardo

    2015-12-01

    The latest achievements in sensor technologies for blood glucose level monitoring, pump miniaturization for insulin delivery, and the availability of portable computing devices are paving the way toward the artificial pancreas as a treatment for diabetes patients. This device encompasses a controller unit that oversees the administration of insulin micro-boluses and continuously drives the pump based on blood glucose readings acquired in real time. In order to foster the research on the artificial pancreas and prepare for its adoption as a therapy, the European Union in 2010 funded the AP@home project, following a series of efforts already ongoing in the USA. This paper, authored by members of the AP@home consortium, reports on the technical issues concerning the design and implementation of an architecture supporting the exploitation of an artificial pancreas platform. First a PC-based platform was developed by the authors to prove the effectiveness and reliability of the algorithms responsible for insulin administration. A mobile-based one was then adopted to improve the comfort for the patients. Both platforms were tested on real patients, and a description of the goals, the achievements, and the major shortcomings that emerged during those trials is also reported in the paper. PMID:25430423

  17. Inhibition of AP-1 by Sulforaphane Involves Interaction with Cysteine in the cFos DNA-Binding Domain; Implications for Chemoprevention of UVB-Induced Skin Cancer

    PubMed Central

    Dickinson, Sally E.; Melton, Tania F.; Olson, Erik R.; Zhang, Jian; Saboda, Kathylynn; Bowden, G. Timothy

    2009-01-01

    Sulforaphane (SFN) is an isothiocyanate derived from cruciferous vegetables which has been linked to decreased risk of certain cancers. Although the role of SFN in the induction of the transcription factor Nrf2 has been studied extensively, there is also evidence that inhibition of the transcription factor AP-1 may contribute to the chemopreventive properties of this compound. In this study, we show for the first time that SFN is effective at reducing the multiplicity and tumor burden of UVB-induced squamous cell carcinomas (SCCs) in a mouse model utilizing co-treatment with the compound and the carcinogen. We also show that SFN pretreatment is able to reduce the activity of AP-1 luciferase in the skin of transgenic mice after UVB. Chromatin immunoprecipitation analysis verified that a main constituent of the AP-1 dimer, cFos, is inhibited from binding to the AP-1 DNA binding site by SFN. EMSA analysis of nuclear proteins also show that SFN and diamide, both known to react with cysteine amino acids, are effective at inhibiting AP-1 from binding to its response element. Using truncated recombinant cFos and cJun we show that mutation of critical cysteines in the DNA binding domain of these proteins (Cys154 in cFos and Cys272 in cJun) results in loss of sensitivity to both SFN and diamide in EMSA analysis. Together, these data indicate that inhibition of AP-1 activity may be an important molecular mechanism in chemoprevention of SCC by SFN. PMID:19671797

  18. Effects of synthetic analogues of poly-APS on contractile response of porcine coronary arteries.

    PubMed

    Grandič, Marjana; Bajuk, Blanka Premrov; Sepčić, Kristina; Košorok, Marinka Drobnič; Frangež, Robert

    2013-03-01

    APS12-2 and APS3 are synthetic analogues of polymeric alkylpyridinium salts (poly-APS) isolated from the marine sponge Reniera sarai. The aim of the present study was to determine the possible direct contractile effects of these two synthetic molecules on coronary arteries, in order partly to explain hemodynamic and cardiotoxic effects of APS12-2 previously observed in in vivo studies and to reveal possible adverse effects on the organism in the case of their clinical use. In contrast to APS3, APS12-2 caused a concentration-dependent vascular smooth muscle contraction of isolated porcine coronary ring preparations in a concentration-range from 1.36 to 13.60μM. Lanthanum chloride (5mM) and verapamil (10μM) completely abolished the APS12-2 evoked contraction of the coronary rings. Pre-incubation with indomethacin (10μM) had no effect on the contractile responses of coronary ring preparations. These results indicate that APS12-2 contracts vascular smooth muscle in a concentration-dependent manner, due to an increase of Ca(2+) influx through the voltage-gated Ca(2+) channels. Our data show for the first time that APS12-2 induces concentration-dependent contraction of coronary ring preparations, which may contribute to the cardiotoxic effects of APS12-2, in addition to hyperkalemia. PMID:23178276

  19. Evolution and protein interactions of AP2 proteins in Brassicaceae: Evidence linking development and environmental responses.

    PubMed

    Zeng, Liping; Yin, Yue; You, Chenjiang; Pan, Qianli; Xu, Duo; Jin, Taijie; Zhang, Bailong; Ma, Hong

    2016-06-01

    Plants have evolved a large number of transcription factors (TF), which are enriched among duplicate genes, highlighting their roles in complex regulatory networks. The APETALA2/EREBP-like genes constitute a large plant TF family and participate in development and stress responses. To probe the conservation and divergence of AP2/EREBP genes, we analyzed the duplication patterns of this family in Brassicaceae and identified interacting proteins of representative Arabidopsis AP2/EREBP proteins. We found that many AP2/EREBP duplicates generated early in Brassicaceae history were quickly lost, but many others were retained in all tested Brassicaceae species, suggesting early functional divergence followed by persistent conservation. In addition, the sequences of the AP2 domain and exon numbers were highly conserved in rosids. Furthermore, we used 16 A. thaliana AP2/EREBP proteins as baits in yeast screens and identified 1,970 potential AP2/EREBP-interacting proteins, with a small subset of interactions verified in planta. Many AP2 genes also exhibit reduced expression in an anther-defective mutant, providing a possible link to developmental regulation. The putative AP2-interacting proteins participate in many functions in development and stress responses, including photomorphogenesis, flower development, pathogenesis, drought and cold responses, abscisic acid and auxin signaling. Our results present the AP2/EREBP evolution patterns in Brassicaceae, and support a proposed interaction network of AP2/EREBP proteins and their putative interacting proteins for further study. PMID:26472270

  20. Identification and proteomic analysis of distinct UBE3A/E6AP protein complexes.

    PubMed

    Martínez-Noël, Gustavo; Galligan, Jeffrey T; Sowa, Mathew E; Arndt, Verena; Overton, Thomas M; Harper, J Wade; Howley, Peter M

    2012-08-01

    The E6AP ubiquitin ligase catalyzes the high-risk human papillomaviruses' E6-mediated ubiquitylation of p53, contributing to the neoplastic progression of cells infected by these viruses. Defects in the activity and the dosage of E6AP are linked to Angelman syndrome and to autism spectrum disorders, respectively, highlighting the need for precise control of the enzyme. With the exception of HERC2, which modulates the ubiquitin ligase activity of E6AP, little is known about the regulation or function of E6AP normally. Using a proteomic approach, we have identified and validated several new E6AP-interacting proteins, including HIF1AN, NEURL4, and mitogen-activated protein kinase 6 (MAPK6). E6AP exists as part of several different protein complexes, including the proteasome and an independent high-molecular-weight complex containing HERC2, NEURL4, and MAPK6. In examining the functional consequence of its interaction with the proteasome, we found that UBE3C (another proteasome-associated ubiquitin ligase), but not E6AP, contributes to proteasomal processivity in mammalian cells. We also found that E6 associates with the HERC2-containing high-molecular-weight complex through its binding to E6AP. These proteomic studies reveal a level of complexity for E6AP that has not been previously appreciated and identify a number of new cellular proteins through which E6AP may be regulated or functioning. PMID:22645313

  1. Induction of neural crest in Xenopus by transcription factor AP2alpha.

    PubMed

    Luo, Ting; Lee, Young-Hoon; Saint-Jeannet, Jean-Pierre; Sargent, Thomas D

    2003-01-21

    We report experiments with Xenopus laevis, using both intact embryos and ectodermal explants, showing that the transcription factor AP2alpha is positively regulated by bone morphogenetic protein (BMP) and Wnt signaling, and that this activation is an essential step in the induction of neural crest (NC). Ectopic expression of AP2alpha is sufficient to activate high-level expression of NC-specific genes such as Slug and Sox9, which can occur as isolated domains within the neural plate as well as by expansion of endogenous NC territories. AP2alpha also has the property of inducing NC in isolated ectoderm in which Wnt signaling is provided but BMP signaling is minimized by overexpression of chordin. Like other NC regulatory factors, activation of AP2alpha requires some attenuation of endogenous BMP signaling; however, this process occurs at a lower threshold for AP2alpha. Furthermore, AP2alpha expression domains are larger than for other NC factors. Loss-of-function experiments with antisense AP2alpha morpholino oligonucleotides result in severe reduction in the NC territory. These results support a central role for AP2alpha in NC induction. We propose a model in which AP2alpha expression, along with inactivation of NC inhibitory factors such as Dlx3, establish a feedback loop comprising AP2alpha, Sox9, and Slug, leading to and maintaining NC specification. PMID:12511599

  2. Expansion and stress responses of AP2/EREBP superfamily in Brachypodium Distachyon

    PubMed Central

    Chen, Lihong; Han, Jiapeng; Deng, Xiaomin; Tan, Shenglong; Li, Lili; Li, Lun; Zhou, Junfei; Peng, Hai; Yang, Guangxiao; He, Guangyuan; Zhang, Weixiong

    2016-01-01

    APETALA2/ethylene-responsive element binding protein (AP2/EREBP) transcription factors constitute one of the largest and most conserved gene families in plant, and play essential roles in growth, development and stress response. Except a few members, the AP2/EREBP family has not been characterized in Brachypodium distachyon, a model plant of Poaceae. We performed a genome-wide study of this family in B. distachyon by phylogenetic analyses, transactivation assays and transcript profiling. A total of 149 AP2/EREBP genes were identified and divided into four subfamilies, i.e., ERF (ethylene responsive factor), DREB (dehydration responsive element binding gene), RAV (related to ABI3/VP) and AP2. Tandem duplication was a major force in expanding B. distachyon AP2/EREBP (BdAP2/EREBP) family. Despite a significant expansion, genomic organizations of BdAP2/EREBPs were monotonous as the majority of them, except those of AP2 subfamily, had no intron. An analysis of transcription activities of several closely related and duplicated BdDREB genes showed their functional divergence and redundancy in evolution. The expression of BdAP2/EREBPs in different tissues and the expression of DREB/ERF subfamilies in B. distachyon, wheat and rice under abiotic stresses were investigated by next-generation sequencing and microarray profiling. Our results are valuable for further function analysis of stress tolerant AP2/EREBP genes in B. distachyon. PMID:26869021

  3. Genome-wide comparison of AP2/ERF superfamily genes between Gossypium arboreum and G. raimondii.

    PubMed

    Lei, Z P; He, D H; Xing, H Y; Tang, B S; Lu, B X

    2016-01-01

    The APETALA2/ethylene response factor (AP2/ERF) transcription factor superfamily is known to regulate diverse processes of plant development and stress responses. We conducted a genome-wide analysis of the AP2/ERF gene in Gossypium arboreum and G. raimondii. Using RPSBLAST and HMMsearch, a total of 271 and 269 AP2/ERF genes were identified in the G. arboreum and G. raimondii genomes, respectively. A phylogenetic analysis classified diploid Gossypium spp AP2/ERF genes into 4 families and 16 subfamilies. Orthologous genes predominated the terminal branch of the phylogenetic tree. Physical mapping showed at least 30% of AP2/ERF genes clustered together. A high level of intra- and inter-species collinearity involving AP2/ERF genes was observed, indicating common (before species divergence) or parallel (after species divergence) segmental duplications, along with tandem duplications, resulting in the species-specific expansion of AP2/ERF genes in diploid Gossypium species. Motif analyses of the AP2/ERF proteins revealed that motif arrangements were highly diverse among subfamilies, but shared by orthologous gene pairs. An examination of nucleotide divergence of AP2/ERF coding regions identified small and non-significant sequence differences among orthologs. Expression profiling of AP2/ERF orthologous gene pairs showed similar abundance levels of orthologous copies between G. arboreum and G. raimondii. Thus, cotton species possess abundant and diverse AP2/ERF genes, resulting from tandem and segmental duplications. Protein and nucleotide sequence and mRNA expression analyses revealed symmetrical evolution, indicating that most AP2/ ERF genes may not have undergone significant biochemical and morphological divergence between sister species. Our study provides detailed insights into the evolutionary characteristics and functional importance of AP2/ERF genes, and could aid in the genetic improvement of agriculturally significant crops in this genus. PMID:27525884

  4. Role of ubiquitin and the HPV E6 oncoprotein in E6AP-mediated ubiquitination

    PubMed Central

    Mortensen, Franziska; Schneider, Daniel; Barbic, Tanja; Sladewska-Marquardt, Anna; Kühnle, Simone; Marx, Andreas; Scheffner, Martin

    2015-01-01

    Deregulation of the ubiquitin ligase E6 associated protein (E6AP) encoded by the UBE3A gene has been associated with three different clinical pictures. Hijacking of E6AP by the E6 oncoprotein of distinct human papillomaviruses (HPV) contributes to the development of cervical cancer, whereas loss of E6AP expression or function is the cause of Angelman syndrome, a neurodevelopmental disorder, and increased expression of E6AP has been involved in autism spectrum disorders. Although these observations indicate that the activity of E6AP has to be tightly controlled, only little is known about how E6AP is regulated at the posttranslational level. Here, we provide evidence that the hydrophobic patch of ubiquitin comprising Leu-8 and Ile-44 is important for E6AP-mediated ubiquitination, whereas it does not affect the catalytic properties of the isolated catalytic HECT domain of E6AP. Furthermore, we show that the HPV E6 oncoprotein rescues the disability of full-length E6AP to use a respective hydrophobic patch mutant of ubiquitin for ubiquitination and that it stimulates E6AP-mediated ubiquitination of Ring1B, a known substrate of E6AP, in vitro and in cells. Based on these data, we propose that E6AP exists in at least two different states, an active and a less active or latent one, and that the activity of E6AP is controlled by noncovalent interactions with ubiquitin and allosteric activators such as the HPV E6 oncoprotein. PMID:26216987

  5. RESULTS OF CESIUM MASS TRANSFER TESTING FOR NEXT GENERATION SOLVENT WITH HANFORD WASTE SIMULANT AP-101

    SciTech Connect

    Peters, T.; Washington, A.; Fink, S.

    2011-09-27

    SRNL has performed an Extraction, Scrub, Strip (ESS) test using the next generation solvent and AP-101 Hanford Waste simulant. The results indicate that the next generation solvent (MG solvent) has adequate extraction behavior even in the face of a massive excess of potassium. The stripping results indicate poorer behavior, but this may be due to inadequate method detection limits. SRNL recommends further testing using hot tank waste or spiked simulant to provide for better detection limits. Furthermore, strong consideration should be given to performing an actual waste, or spiked waste demonstration using the 2cm contactor bank. The Savannah River Site currently utilizes a solvent extraction technology to selectively remove cesium from tank waste at the Multi-Component Solvent Extraction unit (MCU). This solvent consists of four components: the extractant - BoBCalixC6, a modifier - Cs-7B, a suppressor - trioctylamine, and a diluent, Isopar L{trademark}. This solvent has been used to successfully decontaminate over 2 million gallons of tank waste. However, recent work at Oak Ridge National Laboratory (ORNL), Argonne National Laboratory (ANL), and Savannah River National Laboratory (SRNL) has provided a basis to implement an improved solvent blend. This new solvent blend - referred to as Next Generation Solvent (NGS) - is similar to the current solvent, and also contains four components: the extractant - MAXCalix, a modifier - Cs-7B, a suppressor - LIX-79{trademark} guanidine, and a diluent, Isopar L{trademark}. Testing to date has shown that this 'Next Generation' solvent promises to provide far superior cesium removal efficiencies, and furthermore, is theorized to perform adequately even in waste with high potassium concentrations such that it could be used for processing Hanford wastes. SRNL has performed a cesium mass transfer test in to confirm this behavior, using a simulant designed to simulate Hanford AP-101 waste.

  6. AP endonuclease knockdown enhances methyl methanesulfonate hypersensitivity of DNA polymerase β knockout mouse embryonic fibroblasts

    PubMed Central

    Yamamoto, Ryohei; Umetsu, Makio; Yamamoto, Mizuki; Matsuyama, Satoshi; Takenaka, Shigeo; Ide, Hiroshi; Kubo, Kihei

    2015-01-01

    Apurinic/apyrimidinic (AP) endonuclease (Apex) is required for base excision repair (BER), which is the major mechanism of repair for small DNA lesions such as alkylated bases. Apex incises the DNA strand at an AP site to leave 3′-OH and 5′-deoxyribose phosphate (5′-dRp) termini. DNA polymerase β (PolB) plays a dominant role in single nucleotide (Sn-) BER by incorporating a nucleotide and removing 5′-dRp. Methyl methanesulfonate (MMS)-induced damage is repaired by Sn-BER, and thus mouse embryonic fibroblasts (MEFs) deficient in PolB show significantly increased sensitivity to MMS. However, the survival curve for PolB-knockout MEFs (PolBKOs) has a shoulder, and increased sensitivity is only apparent at relatively high MMS concentrations. In this study, we prepared Apex-knockdown/PolB-knockout MEFs (AKDBKOs) to examine whether BER is related to the apparent resistance of PolBKOs at low MMS concentrations. The viability of PolBKOs immediately after MMS treatment was significantly lower than that of wild-type MEFs, but there was essentially no effect of Apex-knockdown on cell viability in the presence or absence of PolB. In contrast, relative counts of MEFs after repair were decreased by Apex knockdown. Parental PolBKOs showed especially high sensitivity at >1.5 mM MMS, suggesting that PolBKOs have another repair mechanism in addition to PolB-dependent Sn-BER, and that the back-up mechanism is unable to repair damage induced by high MMS concentrations. Interestingly, AKDBKOs were hypersensitive to MMS in a relative cell growth assay, suggesting that MMS-induced damage in PolB-knockout MEFs is repaired by Apex-dependent repair mechanisms, presumably including long-patch BER. PMID:25724755

  7. Redox Cycling of Catechol Estrogens Generating Apurinic/Apyrimidinic Sites and 8-oxo-Deoxyguanosine via Reactive Oxygen Species Differentiates Equine and Human Estrogens

    PubMed Central

    Wang, Zhican; Chandrasena, Esala R.; Yuan, Yang; Peng, Kuan-wei; van Breemen, Richard B.; Thatcher, Gregory R. J.; Bolton, Judy L.

    2010-01-01

    Metabolic activation of estrogens to catechols and further oxidation to highly reactive o-quinones generates DNA damage including apurinic/apyrimidinic (AP) sites. 4-Hydroxyequilenin (4-OHEN) is the major catechol metabolite of equine estrogens present in estrogen replacement formulations, known to cause DNA strand breaks, oxidized bases, and stable and depurinating adducts. However, the direct formation of AP sites by 4-OHEN has not been characterized. In the present study, the induction of AP sites in vitro by 4-OHEN and the endogenous catechol estrogen metabolite, 4-hydroxyestrone (4-OHE) was examined by an aldehyde reactive probe assay. Both 4-OHEN and 4-OHE can significantly enhance the levels of AP sites in calf thymus DNA in the presence of the redox cycling agents, copper ion and NADPH. The B-ring unsaturated catechol 4-OHEN induced AP sites without added copper, whereas 4-OHE required copper. AP sites were also generated much more rapidly by 4-OHEN. For both catechol estrogens, the levels of AP sites correlated linearly with 8-oxo-dG levels, implying that depuriniation resulted from reactive oxygen species (ROS) rather than depurination of estrogen-DNA adducts. ROS modulators such as catalase which scavenges hydrogen peroxide and a Cu(I) chelator blocked the formation of AP sites. In MCF-7 breast cancer cells, 4-OHEN significantly enhanced the formation of AP sites with added NADH. In contrast, no significant induction of AP sites was detected in 4-OHE-treated cells. The greater redox activity of the equine catechol estrogen produces rapid oxidative DNA damage via ROS, which is enhanced by redox cycling agents and interestingly by NADPH-dependent quinone oxidoreductase (NQO1). PMID:20509668

  8. The flame structure of AP/HTPB sandwiches

    NASA Astrophysics Data System (ADS)

    Chorpening, Benjamin Todd

    2000-10-01

    Ultraviolet emission imaging experiments have been used to study the combustion of sandwiches of ammonium perchlorate (AP) and hydroxyl-terminated polybutadiene (HTPB) in nitrogen at pressures up to 32 atm, with binder layers from 50 to 450 mum in thickness. An ICCD camera system has been used to image the flame emission near 310 nm, and a backlighting technique has been developed that allows determination of the corresponding surface shape during combustion. The results indicate the AP/HTPB interface regression rate of IPDI cured samples undergoing low power (100W) laser-assisted deflagration is nearly independent of the binder thickness for binders thicker than 100 mum. The pressure exponent of the regression rate is 0.31 up to 15 atm, increasing with pressure from 15 to 32 atm. Two primary regimes of flame behavior have been identified: a split flame base regime which occurs with high Peclet and Damkohler numbers, and a merged flame base regime which occurs with low Peclet and Damkohler numbers. A secondary regime, exhibiting a "lifted" flame, occurs with low Damkohler numbers and high Peclet numbers. The ultraviolet flame emissions observed in the experiments show a correspondence with the fuel-rich region of the flame, as determined with a Schvab-Zeldovich model. This is reasonable since the primary sources of ultraviolet emission in the 305--315 nm region, electronically excited OH and the CO + O reaction, are dependent on fuel related species. The growth of the fuel-rich region with increasing Peclet number, predicted by the model, is qualitatively matched by the experimental results. The predicted shrinkage of the fuel-rich region when the binder layer is diluted with fine AP is also qualitatively matched by the experiments. Comparison of the experimental results with a single-reaction model with finite rate kinetics shows a weak qualitative agreement on the influence of Damkohler number. A large increase in Damkohler number (factor of 20) leads to a strong

  9. A CRE/AP-1-Like Motif Is Essential for Induced Syncytin-2 Expression and Fusion in Human Trophoblast-Like Model

    PubMed Central

    Vargas, Amandine; Rassart, Éric; Barbeau, Benoit

    2015-01-01

    Syncytin-2 is encoded by the envelope gene of Endogenous Retrovirus-FRD (ERVFRD-1) and plays a critical role in fusion of placental trophoblasts leading to the formation of the multinucleated syncytiotrophoblast. Its expression is consequently regulated in a strict manner. In the present study, we have identified a forskolin-responsive region located between positions -300 to -150 in the Syncytin-2 promoter region. This 150 bp region in the context of a minimal promoter mediated an 80-fold induction of promoter activity following forskolin stimulation. EMSA analyses with competition experiments with nuclear extracts from forskolin-stimulated BeWo cells demonstrated that the -211 to -177 region specifically bound two forskolin-induced complexes, one of them containing a CRE/AP-1-like motif. Site-directed mutagenesis of the CRE/AP-1 binding site in the context of the Syncytin-2 promoter or a heterologous promoter showed that this motif was mostly essential for forskolin-induced promoter activity. Transfection experiments with dominant negative mutants and constitutively activated CREB expression vectors in addition to Chromatin Immunoprecipitation suggested that a CREB family member, CREB2 was binding and acting through the CRE/AP-1 motif. We further demonstrated the binding of JunD to this same motif. Similar to forskolin and soluble cAMP, CREB2 and JunD overexpression induced Syncytin-2 promoter activity in a CRE/AP-1-dependent manner and Syncytin-2 expression. In addition, BeWo cell fusion was induced by both CREB2 and JunD overexpression, while being repressed following silencing of either gene. These results thereby demonstrate that induced expression of Syncytin-2 is highly dependent on the interaction of bZIP-containing transcription factors to a CRE/AP-1 motif and that this element is important for the regulation of Syncytin-2 expression, which results in the formation of the peripheral syncytiotrophoblast layer. PMID:25781974

  10. A CD2AP Mutation Associated with Focal Segmental Glomerulosclerosis in Young Adulthood

    PubMed Central

    Tsvetkov, Dmitry; Hohmann, Michael; Anistan, Yoland Marie; Mannaa, Marwan; Harteneck, Christian; Rudolph, Birgit; Gollasch, Maik

    2016-01-01

    Mutations in CD2-associated protein (CD2AP) have been identified in patients with focal segmental glomerulosclerosis (FSGS); however, reports of CD2AP mutations remain scarce. We performed Sanger sequencing in a patient with steroid-resistant FSGS and identified a heterozygous CD2AP mutation (p.T374A, c.1120 A > G). Our patient displayed mild cognitive decline, a phenotypic characteristic not previously associated with CD2AP-associated FSGS. His proteinuria was remarkably reduced by treatment with cyclosporine A. Our findings expand the genetic spectrum of CD2AP-associated disorders and broaden the associated phenotype with the co-occurrence of cognitive decline. Our case shows that cyclosporin A is a treatment option for CD2AP-associated nephropathy. PMID:26997877

  11. A CD2AP Mutation Associated with Focal Segmental Glomerulosclerosis in Young Adulthood.

    PubMed

    Tsvetkov, Dmitry; Hohmann, Michael; Anistan, Yoland Marie; Mannaa, Marwan; Harteneck, Christian; Rudolph, Birgit; Gollasch, Maik

    2016-01-01

    Mutations in CD2-associated protein (CD2AP) have been identified in patients with focal segmental glomerulosclerosis (FSGS); however, reports of CD2AP mutations remain scarce. We performed Sanger sequencing in a patient with steroid-resistant FSGS and identified a heterozygous CD2AP mutation (p.T374A, c.1120 A > G). Our patient displayed mild cognitive decline, a phenotypic characteristic not previously associated with CD2AP-associated FSGS. His proteinuria was remarkably reduced by treatment with cyclosporine A. Our findings expand the genetic spectrum of CD2AP-associated disorders and broaden the associated phenotype with the co-occurrence of cognitive decline. Our case shows that cyclosporin A is a treatment option for CD2AP-associated nephropathy. PMID:26997877

  12. The Forkhead Transcription Factor FOXK2 Promotes AP-1-Mediated Transcriptional Regulation

    PubMed Central

    Ji, Zongling; Donaldson, Ian J.; Liu, Jingru; Hayes, Andrew; Zeef, Leo A. H.

    2012-01-01

    The transcriptional control circuitry in eukaryotic cells is complex and is orchestrated by combinatorially acting transcription factors. Forkhead transcription factors often function in concert with heterotypic transcription factors to specify distinct transcriptional programs. Here, we demonstrate that FOXK2 participates in combinatorial transcriptional control with the AP-1 transcription factor. FOXK2 binding regions are widespread throughout the genome and are often coassociated with AP-1 binding motifs. FOXK2 acts to promote AP-1-dependent gene expression changes in response to activation of the AP-1 pathway. In this context, FOXK2 is required for the efficient recruitment of AP-1 to chromatin. Thus, we have uncovered an important new molecular mechanism that controls AP-1-dependent gene expression. PMID:22083952

  13. The beta-appendages of the four adaptor-protein (AP) complexes: structure and binding properties, and identification of sorting nexin 9 as an accessory protein to AP-2.

    PubMed Central

    Lundmark, Richard; Carlsson, Sven R

    2002-01-01

    Adaptor protein (AP) complexes are essential components for the formation of coated vesicles and the recognition of cargo proteins for intracellular transport. Each AP complex exposes two appendage domains with that function to bind regulatory accessory proteins in the cytosol. Secondary structure predictions, sequence alignments and CD spectroscopy were used to relate the beta-appendages of all human AP complexes to the previously published crystal structure of AP-2. The results suggested that the beta-appendages of AP-1, AP-2 and AP-3 have similar structures, consisting of two subdomains, whereas that of AP-4 lacks the inner subdomain. Pull-down and overlay assays showed partial overlap in the binding specificities of the beta-appendages of AP-1 and AP-2, whereas the corresponding domain of AP-3 displayed a unique binding pattern. That AP-4 may have a truncated, non-functional domain was indicated by its apparent inability to bind any proteins from cytosol. Of several novel beta-appendage-binding proteins detected, one that had affinity exclusively for AP-2 was identified as sorting nexin 9 (SNX9). SNX9, which contains a phox and an Src homology 3 domain, was found in large complexes and was at least partially associated with AP-2 in the cytosol. SNX9 may function to assist AP-2 in its role at the plasma membrane. PMID:11879186

  14. APS storage ring commissioning and early operational experience

    SciTech Connect

    Decker, G.

    1995-07-01

    The Advanced Photon Source (APS) at Argonne National Laboratory (ANL) uses a 100-mA, 7-GeV positron storage ring to produce high brilliance bending magnet and insertion device x-rays for up to 70 x-ray beamlines. It is 1104 meters in circumference and has a beam liftime designed to exceed 10 hours with 1 nTorr average ring vacuum at 100 mA. The high brilliance required by the synchrotron light users results from the storage ring`s natural emittance of 8.2 nm-rad, together with the requirement that the beam be stable to a level which is less than 5% of its rms size. Real-time closed orbit feedback is employed to achieve the required stability and is discussed elsewhere in these proceedings. Installation of storage ring components was completed early this year, and we report here on the first experiences of commissioning and operation with beam.

  15. Commissioning results of the APS storage ring diagnostics systems

    SciTech Connect

    Lumpkin, A.H.

    1996-12-31

    Initial commissionings of the Advanced Photon Source (APS) 7-GeV storage ring and its diagnostics systems have been done. Early studies involved single-bunch measurements for beam transverse size ({sigma}{sub x} {approx} 150 {mu}m, {sigma}{sub y} {approx} 50 {mu}m), current, injection losses, and bunch length. The diagnostics have been used in studies related to the detection of an extra contribution to beam jitter at {approximately} 6.5 Hz frequency; observation of bunch lengthening ({sigma} {approx} 30 to 60 ps) with single-bunch current; observation of an induced vertical, head-tail instability; and detection of a small orbit change with insertion device gap position. More recently, operations at 100-mA stored-beam current, the baseline design goal, have been achieved with the support of beam characterizations.

  16. The chemical abundances of the Ap star HD94660

    SciTech Connect

    Giarrusso, M.

    2014-05-09

    In this work I present the determination of chemical abundances of the Ap star HD94660, a possible rapid oscillating star. As all the magnetic chemically peculiar objects, it presents CNO underabundance and overabundance of iron peak elements of ∼100 times and of rare earths up to 4 dex with respect to the Sun. The determination was based on the conversion of the observed equivalent widths into abundances simultaneously to the determination of effective temperature and gravity. Since the Balmer lines of early type stars are very sensitive to the surface gravity while the flux distribution is sensitive to the effective temperature, I have adopted an iterative procedure to match the H{sub α} line profile and the observed UV-Vis-NIR magnitudes of HD94660 looking for a consistency between the metallicity of the atmosphere model and the derived abundances. From my spectroscopic analysis, this star belongs to the no-rapid oscillating class.

  17. Thermal analysis of the crotch absorber in APS

    SciTech Connect

    Sheng, I.C.; Howell, J.

    1992-10-01

    A crotch absorber design for use in the Advanced Photon source (APS) has been proposed and analyzed. the absorber is placed downstream of sectors S2 and S4 in the curved storage ring chamber and will be subjected to a peak power of 120 W/mm{sup 2} per 100mA synchrotron radiation. A beryllium ring is brazed on the GlidCop cooling cylinder in order to diffuse the concentrated bending magnet heating. One concentric water channel and two annular return water channels are arranged in the GlidCop cylinder to enhance the cooling. A Bodner-Partom thermoviscoplastic constitutive equation and a modified Manson-Coffin fatigue relation are proposed to simulate the cyclic thermal loading, as well as to predict the thermal fatigue life of the crotch absorber. Results of temperature and stress using finite element computations are displayed and series of e-beam welder tests and microstructure measurements are reported.

  18. The origin of light variability in Ap stars

    NASA Astrophysics Data System (ADS)

    Krtička, J.; Mikulášek, Z.; Zverko, J.; Prvák, M.

    2014-11-01

    The usual photometric variability detected in Ap stars is that associated with rotation. It has long been surmised that redistribution of rotationally modulated flux in surface abundance spots is the cause of that type of variability. The redistribution is caused by bound-bound (line) and bound-free (continuum) transitions of various elements, in particular helium, silicon, iron, and (obviously) also rare-earth elements. With the availability of detailed abundance maps, complete atomic data, and detailed model atmospheres it has now become possible to simulate reliably the photometric variability due to rotation. This is demonstrated by the example of several CP stars. We generalise these result and discuss the importance of individual elements in terms of their dependence on effective temperature. We emphasise the importance of light curve prediction for testing surface abundance maps and the atomic data.

  19. Thermal analysis of the crotch absorber in APS

    SciTech Connect

    Sheng, I.C.; Howell, J.

    1992-01-01

    A crotch absorber design for use in the Advanced Photon source (APS) has been proposed and analyzed. the absorber is placed downstream of sectors S2 and S4 in the curved storage ring chamber and will be subjected to a peak power of 120 W/mm{sup 2} per 100mA synchrotron radiation. A beryllium ring is brazed on the GlidCop cooling cylinder in order to diffuse the concentrated bending magnet heating. One concentric water channel and two annular return water channels are arranged in the GlidCop cylinder to enhance the cooling. A Bodner-Partom thermoviscoplastic constitutive equation and a modified Manson-Coffin fatigue relation are proposed to simulate the cyclic thermal loading, as well as to predict the thermal fatigue life of the crotch absorber. Results of temperature and stress using finite element computations are displayed and series of e-beam welder tests and microstructure measurements are reported.

  20. Nomenclature and name assignment rules for the APS storage ring

    SciTech Connect

    Decker, G.

    1992-03-16

    Because the APS accelerators are moving into the fabrication/assembly/installation stage, it is important for consistent naming conventions to be used throughout the project. The intent of this note is to dictate the rules to be adhered to when naming devices in the storage ring. These rules are generic in nature, and shall be applied in principle to the other machines as well. It is essential that every component have a unique and, hopefully, easily recognizable name. Every ASD and XFD group, except for magnets, must interface with the control system. For this reason all device names were developed keeping in mind their actual function, such as controlling or monitoring some device in the ring. Even though magnets are not directly interfaced to the control system, their power supplies are; therefore, a magnet will have the same name as its associated power supply.

  1. Klystron modulator operation and upgrades for the APS linac

    SciTech Connect

    Russell, T.J.; Cours, A.

    1995-07-01

    The Advanced Photon Source (APS) linac requires five 100-MW modulators to achieve its required energy. In-house construction of these modulators was under an extremely compressed time schedule and, while the original design was successful, it had a few shortcomings. The operation of the modulators was hindered by excessively sensitive controls and overheating during the hot summer months. The system underwent minor changes that resulted in major improvements. Additionally, improvements have been made to the high voltage circuits to improve the rise time of the output pulse shape. reduce the initial ringing of the pulse, and enhance the reliability of the system. This paper will outline the changes and explain the results of the improvements.

  2. The APS intranet as a man-machine interface.

    SciTech Connect

    Ciarlette, D.; Gerig, R.; McDowell, W.

    1997-12-02

    The Advanced Photon Source at Argonne National Laboratory has implemented a number of methods for people to interact with the accelerator systems. The accelerator operators use Sun workstations running MEDM and WCL to interface interactively with the accelerator, however, many people need to view information rather than interact with the machine. One of the most common interfaces for viewing information at the Advanced Photon Source is the World Wide Web. Information such as operations logbook entries, machine status updates, and displays of archived and current data are easily available to APS personnel. This interface between people and the accelerator has proven to be quite useful. Because the Intranet is operating-system independent and inherently unidirectional, ensuring the prevention of unauthorized or accidental control of the accelerators is straightforward.

  3. Some considerations in the combustion of AP/composite propellants

    NASA Technical Reports Server (NTRS)

    Kumar, R. N.

    1972-01-01

    Theoretical studies are presented on the time-independent and oscillatory combustion of nonmetallized AP/composite propellants. Three hypotheses are introduced: (1) The extent of propellant degradation at the vaporization step has to be specified through a scientific criterion. (2) The condensed phase degradation reaction of ammonium perchlorate to a vaporizable state is the overall rate-limiting step. (3) Gas phase combustion rate is controlled by the mixing rate of fuel and oxidizer vapors. In the treatment of oscillatory combustion, the assumption of quasi-steady fluctuations in the gas phase is used to supplement these hypotheses. In comparison with experimental data, this study predicts several of the observations including a few that remain inconsistent with theoretical results.

  4. Aerosol Polarimetry Sensor (APS): Design Summary, Performance and Potential Modifications

    NASA Technical Reports Server (NTRS)

    Cairns, Brian

    2014-01-01

    APS is a mature design that has already been built and has a TRL of 7. Algorithmic and retrieval capabilities continue to improve and make better and more sophisticated used of the data. Adjoint solutions, both in one dimensional and three dimensional are computationally efficient and should be the preferred implementation for the calculation of Jacobians (one dimensional), or cost-function gradients (three dimensional). Adjoint solutions necessarily provide resolution of internal fields and simplify incorporation of active measurements in retrievals, which will be necessary for a future ACE mission. Its best to test these capabilities when you know the answer: OSSEs that are well constrained observationally provide the best place to test future multi-instrument platform capabilities and ensure capabilities will meet scientific needs.

  5. Fabrication and Testing of Deflecting Cavities for APS

    SciTech Connect

    Mammosser, John; Wang, Haipeng; Rimmer, Robert; Jim, Henry; Katherine, Wilson; Dhakal, Pashupati; Ali, Nassiri; Jim, Kerby; Jeremiah, Holzbauer; Genfa, Wu; Joel, Fuerst; Yawei, Yang; Zenghai, Li

    2013-09-01

    Jefferson Lab (Newport News, Virginia) in collaboration with Argonne National Laboratory (Argonne, IL) has fabricated and tested four first article, 2.8 GHz, deflecting SRF cavities, for Argonne's Short-Pulse X-ray (SPX) project. These cavities are unique in many ways including the fabrication techniques in which the cavity cell and waveguides were fabricated. These cavity subcomponents were milled from bulk large grain niobium ingot material directly from 3D CAD files. No forming of sub components was used with the exception of the beam-pipes. The challenging cavity and helium vessel design and fabrication results from the stringent RF performance requirements required by the project and operation in the APS ring. Production challenges and fabrication techniques as well as testing results will be discussed in this paper.

  6. Dropping of mixing pump in Tank 102-AP

    SciTech Connect

    Jimenez, R.F.

    1995-06-02

    The purpose of this study is to examine dropping of the mixing pump in Tank 102-AP during its removal poses the risk of causing a leak in the tank bottom with attendant potential for public exposure from the leak. The purpose of this investigation is to examine the potential for causing such a leak (i.e., estimated frequency of leak occurrence); to qualitatively estimate leak magnitude if its is a credible event; and, finally to compare the worker hazard, in the installation of an impact limiter (should it be required), to that which the public might incur if a leak is manifest in the tank bottom. The ultimate goal of the study is, of course, to assess the need for installation of an impact limiter.

  7. Global coupling and decoupling of the APS storage ring

    SciTech Connect

    Chae, Y.C.; Liu, J.; Teng, L.C.

    1993-07-01

    This paper describes a study of controlling the coupling between the horizontal and the vertical betatron oscillations in the 7-GeV Advanced Photon Source (APS) storage ring. First, we investigate the strengthening of coupling using two families of skew quadrupoles. Twenty skew quadrupoles are arranged in the 40 sectors of the storage ring and powered in such a way so as to generate both quadrature components of the required 21st harmonic. The numerical results from tracking a single particle are presented for the various configurations of skew quadrupoles. Second, we describe the global decoupling procedure to minimize the unwanted coupling effects. These are mainly due to the random roll errors of normal quadruples. It is shown that even with the rather large rms roll error of 2 mrad, the coupling effects can be compensated for with 20 skew quadrupoles each having maximum strength one order of magnitude lower than the typical normal quadrupole strength.

  8. Improved temperature regulation of APS linac RF components.

    SciTech Connect

    Dortwegt, R.

    1998-09-21

    The temperature of the APS S-Band linac's high-power rf components is regulated by water from individual closed-loop deionized (DI) water systems. The rf components are all made of oxygen-free high-conductivity copper and respond quickly to temperature changes. The SLED cavities are especially temperature-sensitive and cause beam energy instabilities when the temperature is not well regulated. Temperature regulation better than {+-} 0.1 F is required to achieve good energy stability. Improvements in the closed-loop water systems have enabled them to achieve a regulation of {+-} 0.05 F over long periods. Regulation philosophy and equipment are discussed and numerical results are presented.

  9. Progress on the development of APS beam position monitoring system

    SciTech Connect

    Decker, G.; Chung, Youngjoo.

    1991-01-01

    This paper describes the development status of the beam position monitoring system for the Advanced Photon Source (APS), a third-generation light source now under construction at Argonne National Laboratory. The accelerator complex will consist of an electron linac, a positron linac, a positron accumulator ring (PAR), an injector synchrotron and a storage ring. For beam position measurement, striplines will be used on the linacs, while button-type pickups will be used on the injector synchrotron and the storage ring. A test stand with a prototype injector synchrotron beam position monitor (BPM) unit has been built, and we present the results of position calibration measurements using a wire. Comparison of the results with theoretical calculations will be presented. The current effort on similar storage ring BPM system measurements will also be discussed. 4 refs., 5 figs., 2 tabs.

  10. A new look at AP/composite propellant combustion

    NASA Technical Reports Server (NTRS)

    Kumar, R. N.

    1973-01-01

    Some theoretical studies on the time-independent and oscillatory combustion of nonmetallized ammonium perchlorate (AP)/composite propellants are presented. A coherent and unified interpretation was made of the voluminous data available from experiments related to propellant combustion. Three fundamental hypotheses are introduced: the extent of propellant degradation at the vaporization step has to be specified through a scientific criterion; the condensed-phase degradation reaction of ammonium perchlorate to a vaporizable state is the overall rate-limiting step; gas-phase combustion rate is controlled by the mixing rate of fuel and oxidizer vapors. In the treatment of oscillatory combustion, the assumption of quasi-steady fluctuations in the gas phase is used to supplement these hypotheses.

  11. The History of the APS Shock Compression of Condensed Matter Topical Group

    NASA Astrophysics Data System (ADS)

    Forbes, Jerry W.

    2001-06-01

    To provide broader scientific recognition and to advance the science of shock-compressed condensed matter, a group of APS members worked within the Society to make this technical field an active part of APS. Individual papers were given at APS meetings starting in the 1950’s and then later whole sessions were organized starting at the 1967 Pasadena meeting. Topical conferences began in 1979 in Pullman, WA where George Duvall and Dennis Hayes were co-chairs. Most all early topical conferences were sanctioned by the APS while those held after 1985 were official APS meetings. In 1984, after consulting with a number of people in the shock wave field, Robert Graham circulated a petition to form an APS topical group. He obtained signatures from a balanced cross-section of the community. William Havens, the executive secretary of APS, informed Robert Graham by letter on November 28, 1984 that the APS Council had officially accepted the formation of this topical group at its October 28, 1984 meeting. The first election occurred July 23, 1985 where Robert Graham was elected chairman, William Nellis vice-chairman, and Jerry Forbes secretary/treasurer. The topical group remains viable today by holding a topical conference in odd numbered years and shock wave sessions at APS general meetings in even numbered years A major benefit of being an official unit of APS is the allotment of APS fellows every year. The APS shock compression award established in 1987, has also provided broad recognition of many major scientific accomplishments in this field.

  12. HPV16 E6 and E6AP differentially cooperate to stimulate or augment Wnt signaling

    SciTech Connect

    Sominsky, Sophia; Kuslansky, Yael; Shapiro, Beny; Jackman, Anna; Haupt, Ygal; Rosin-Arbesfeld, Rina; Sherman, Levana

    2014-11-15

    The present study investigated the roles of E6 and E6AP in the Wnt pathway. We showed that E6 levels are markedly reduced in cells in which Wnt signaling is activated. Coexpression of wild-type or mutant E6AP (C820A) in Wnt-activated cells stabilized E6 and enhanced Wnt/β-catenin/TCF transcription. Expression of E6AP alone in nonstimulated cells elevated β-catenin level, promoted its nuclear accumulation, and activated β-catenin/TCF transcription. A knockdown of E6AP lowered β-catenin levels. Coexpression with E6 intensified the activities of E6AP. Further experiments proved that E6AP/E6 stabilize β-catenin by protecting it from proteasomal degradation. This function was dependent on the catalytic activity of E6AP, the kinase activity of GSK3β and the susceptibility of β-catenin to GSK3β phosphorylation. Thus, this study identified E6AP as a novel regulator of the Wnt signaling pathway, capable of cooperating with E6 in stimulating or augmenting Wnt/β-catenin signaling, thereby possibly contributing to HPV carcinogenesis. - Highlights: • The roles of E6 and E6AP in the Wnt pathway were investigated. • E6AP stabilizes E6 and enhances E6 activity in augmentation of Wnt signaling. • E6AP cooperates with E6 to stabilize β-catenin and stimulate Wnt/β-catenin signaling. • E6AP and E6 act through different mechanisms to augment or stimulate Wnt signaling.

  13. CREB Inhibits AP-2α Expression to Regulate the Malignant Phenotype of Melanoma

    PubMed Central

    Zigler, Maya; Villares, Gabriel J.; Braeuer, Russell R.; Wang, Hua; Huang, Li; Bar-Eli, Menashe

    2010-01-01

    Background The loss of AP-2α and increased activity of cAMP-responsive element binding (CREB) protein are two hallmarks of malignant progression of cutaneous melanoma. However, the molecular mechanism responsible for the loss of AP-2α during melanoma progression remains unknown. Methodology/Principal Findings Herein, we demonstrate that both inhibition of PKA-dependent CREB phosphorylation, as well as silencing of CREB expression by shRNA, restored AP-2α protein expression in two metastatic melanoma cell lines. Moreover, rescue of CREB expression in CREB-silenced cell lines downregulates expression of AP-2α. Loss of AP-2α expression in metastatic melanoma occurs via a dual mechanism involving binding of CREB to the AP-2α promoter and CREB-induced overexpression of another oncogenic transcription factor, E2F-1. Upregulation of AP-2α expression following CREB silencing increases endogenous p21Waf1 and decreases MCAM/MUC18, both known to be downstream target genes of AP-2α involved in melanoma progression. Conclusions/Significance Since AP-2α regulates several genes associated with the metastatic potential of melanoma including c-KIT, VEGF, PAR-1, MCAM/MUC18, and p21Waf1, our data identified CREB as a major regulator of the malignant melanoma phenotype. PMID:20805990

  14. AP-2β is a transcriptional regulator for determination of digit length in tetrapods.

    PubMed

    Seki, Ryohei; Kitajima, Keiichi; Matsubara, Haruka; Suzuki, Takayuki; Saito, Daisuke; Yokoyama, Hitoshi; Tamura, Koji

    2015-11-01

    The species-specific morphology of digits in the tetrapod limb, including the length and number of metacarpal, metatarsal, and phalangeal bones, suggests that a common developmental mechanism for digit formation is modified in a species-specific manner. Here, we examined the function of the AP-2β transcription factor in regulating digit length in the chicken autopod. Mutations in the gene encoding AP-2β are associated with Char syndrome, a human autosomal dominant disorder. Char syndrome patients exhibit autopod skeletal defects, including loss of phalanges and shortened fingers, suggestive of a function for AP-2β in normal digit development. The ectopic expression of two different dominant-negative forms of chick AP-2β, equivalent to mutant forms associated with human Char syndrome, in the developing chick hindlimb bud resulted in defective digit formation, including reductions in the number and length of phalanges and metatarsals. A detailed analysis of the AP-2β expression pattern in the limb bud indicated a correlation between the pattern/duration of AP-2β expression in the limb mesenchyme and digit length in three amniote species, the chicken, mouse and gecko. In addition, we found that AP-2β expression was downstream of Fgf signals from the apical ectodermal ridge, which is crucial in digit morphogenesis, and that excessive AP-2β function resulted in dysregulated digit length. Taken together, these results suggest that AP-2β functions as a novel transcriptional regulator for digit morphogenesis. PMID:26277217

  15. Genome-wide analysis of the AP2/ERF gene family in Salix arbutifolia

    PubMed Central

    Rao, Guodong; Sui, Jinkai; Zeng, Yanfei; He, Caiyun; Zhang, Jianguo

    2015-01-01

    AP2/ERF genes encode transcriptional regulators with a variety of functions in plant growth and development and in response to biotic and abiotic stresses. To date, there are no detailed classification and expression profiles for AP2/ERF genes in Salix. In this study, a comprehensive computational analysis identified 173 AP2/ERF superfamily genes in willow (Salix arbutifolia), by using in silico cloning methods with the use of the AP2/ERF conserved domain amino acid sequence of Arabidopsis thaliana as a probe. Based on the results of phylogenetic analyses and the number of AP2/ERF domains, the AP2/ERF genes were classified into four groups: AP2, RAV, ERF and Soloist. The expression profile was analyzed using transcriptome data from different tissues. A comparative analysis of AP2/ERF superfamily genes among Salix, Populus and Arabidopsis was performed. The Salix DREB, AP2 and RAV groups had a similar number to those in Arabidopsis, and the size of the ERF subfamily in Salix was about 1.4-fold that of Arabidopsis. The Salix DREB subfamily was smaller compared to Populus, while the other families were similar in size to those in Populus. These results will be useful for future functional analyses of the ERF family genes. PMID:25830086

  16. Acyl-CoA synthetase catalyzes the synthesis of diadenosine hexaphosphate (Ap6A).

    PubMed

    Fontes, R; Günther Sillero, M A; Sillero, A

    1999-03-01

    The synthesis of diadenosine hexaphosphate (Ap6A), a potent vasoconstrictor, is catalyzed by acyl-CoA synthetase from Pseudomonas fragi. In a first step AMP is transferred from ATP to tetrapolyphosphate (P4) originating adenosine pentaphosphate (p5A) which, subsequently, is the acceptor of another AMP moiety from ATP generating diadenosine hexaphosphate (Ap6A). Diadenosine pentaphosphate (Ap5A) and diadenosine tetraphosphate (Ap4A) were also synthesized in the course of the reaction. In view of the variety of biological effects described for these compounds the potential capacity of synthesis of diadenosine polyphosphates by the mammalian acyl-CoA synthetases may be relevant. PMID:10385004

  17. Isolation, classification and transcription profiles of the AP2/ERF transcription factor superfamily in citrus.

    PubMed

    Xie, Xiu-lan; Shen, Shu-ling; Yin, Xue-ren; Xu, Qian; Sun, Chong-de; Grierson, Donald; Ferguson, Ian; Chen, Kun-song

    2014-07-01

    The AP2/ERF gene family encodes plant-specific transcription factors. In model plants, AP2/ERF genes have been shown to be expressed in response to developmental and environmental stimuli, and many function downstream of the ethylene, biotic, and abiotic stress signaling pathways. In citrus, ethylene is effective in regulation citrus fruit quality, such as degreening and aroma. However, information about the citrus AP2/ERF family is limited, and would enhance our understanding of fruit responses to environmental stress, fruit development and quality. CitAP2/ERF genes were isolated using the citrus genome database, and their expression patterns analyzed by real-time PCR using various orange organs and samples from a fruit developmental series. 126 sequences with homologies to AP2/ERF proteins were identified from the citrus genome, and, on the basis of their structure and sequence, assigned to the ERF family (102), AP2 family (18), RAV family (4) and Soloist (2). MEME motif analysis predicted the defining AP2/ERF domain and EAR repressor domains. Analysis of transcript accumulation in Citrus sinensis cv. 'Newhall' indicated that CitAP2/ERF genes show organ-specific and temporal expression, and provided a framework for understanding the transcriptional regulatory roles of AP2/ERF gene family members in citrus. Hierarchical cluster analysis and t tests identified regulators that potentially function during orange fruit growth and development. PMID:24566692

  18. The AP-3 adaptor complex is required for vacuolar function in Arabidopsis

    PubMed Central

    Zwiewka, Marta; Feraru, Elena; Möller, Barbara; Hwang, Inhwan; Feraru, Mugurel I; Kleine-Vehn, Jürgen; Weijers, Dolf; Friml, Jiří

    2011-01-01

    Subcellular trafficking is required for a multitude of functions in eukaryotic cells. It involves regulation of cargo sorting, vesicle formation, trafficking and fusion processes at multiple levels. Adaptor protein (AP) complexes are key regulators of cargo sorting into vesicles in yeast and mammals but their existence and function in plants have not been demonstrated. Here we report the identification of the protein-affected trafficking 4 (pat4) mutant defective in the putative δ subunit of the AP-3 complex. pat4 and pat2, a mutant isolated from the same GFP imaging-based forward genetic screen that lacks a functional putative AP-3 β, as well as dominant negative AP-3 μ transgenic lines display undistinguishable phenotypes characterized by largely normal morphology and development, but strong intracellular accumulation of membrane proteins in aberrant vacuolar structures. All mutants are defective in morphology and function of lytic and protein storage vacuoles (PSVs) but show normal sorting of reserve proteins to PSVs. Immunoprecipitation experiments and genetic studies revealed tight functional and physical associations of putative AP-3 β and AP-3 δ subunits. Furthermore, both proteins are closely linked with putative AP-3 μ and σ subunits and several components of the clathrin and dynamin machineries. Taken together, these results demonstrate that AP complexes, similar to those in other eukaryotes, exist in plants, and that AP-3 plays a specific role in the regulation of biogenesis and function of vacuoles in plant cells. PMID:21670741

  19. AP Selection Algorithm for Real-Time Communications through Mixed WLAN Environments

    NASA Astrophysics Data System (ADS)

    Morioka, Yasufumi; Higashino, Takeshi; Tsukamoto, Katsutoshi; Komaki, Shozo

    Recent rapid development of high-speed wireless access technologies has created mixed WLAN (Wireless LAN) environments where QoS capable APs coexist with legacy APs. To provide QoS guarantee in this mixed WLAN environment, this paper proposes a new AP selection algorithm. The proposed algorithm assigns an STA (Station) to an AP in the overall WLAN service area. Simulation results show improvement in the VoIP performance in terms of an eMOS (estimated Mean Opinion Score) value and the FTP throughput compared to conventional algorithms.

  20. Mutations in the gene encoding the Sigma 2 subunit of the adaptor protein 1 complex, AP1S2, cause X-linked mental retardation.

    PubMed

    Tarpey, Patrick S; Stevens, Claire; Teague, Jon; Edkins, Sarah; O'Meara, Sarah; Avis, Tim; Barthorpe, Syd; Buck, Gemma; Butler, Adam; Cole, Jennifer; Dicks, Ed; Gray, Kristian; Halliday, Kelly; Harrison, Rachel; Hills, Katy; Hinton, Jonathon; Jones, David; Menzies, Andrew; Mironenko, Tatiana; Perry, Janet; Raine, Keiran; Richardson, David; Shepherd, Rebecca; Small, Alexandra; Tofts, Calli; Varian, Jennifer; West, Sofie; Widaa, Sara; Yates, Andy; Catford, Rachael; Butler, Julia; Mallya, Uma; Moon, Jenny; Luo, Ying; Dorkins, Huw; Thompson, Deborah; Easton, Douglas F; Wooster, Richard; Bobrow, Martin; Carpenter, Nancy; Simensen, Richard J; Schwartz, Charles E; Stevenson, Roger E; Turner, Gillian; Partington, Michael; Gecz, Jozef; Stratton, Michael R; Futreal, P Andrew; Raymond, F Lucy

    2006-12-01

    In a systematic sequencing screen of the coding exons of the X chromosome in 250 families with X-linked mental retardation (XLMR), we identified two nonsense mutations and one consensus splice-site mutation in the AP1S2 gene on Xp22 in three families. Affected individuals in these families showed mild-to-profound mental retardation. Other features included hypotonia early in life and delay in walking. AP1S2 encodes an adaptin protein that constitutes part of the adaptor protein complex found at the cytoplasmic face of coated vesicles located at the Golgi complex. The complex mediates the recruitment of clathrin to the vesicle membrane. Aberrant endocytic processing through disruption of adaptor protein complexes is likely to result from the AP1S2 mutations identified in the three XLMR-affected families, and such defects may plausibly cause abnormal synaptic development and function. AP1S2 is the first reported XLMR gene that encodes a protein directly involved in the assembly of endocytic vesicles. PMID:17186471

  1. Exploring the mechanism of how tvMyb2 recognizes and binds ap65-1 by molecular dynamics simulations and free energy calculations.

    PubMed

    Li, Wei-Kang; Zheng, Qing-Chuan; Zhang, Hong-Xing

    2016-01-01

    TvMyb2, one of the Myb-like transcriptional factors in Trichomonas vaginalis, binds to two closely spaced promoter sites, MRE-1/MRE-2r and MRE-2f, on the ap65-1 gene. However, detailed dynamical structural characteristics of the tvMyb2-ap65-1 complex and a detailed study of the protein in the complex have not been done. Focused on a specific tvMyb2-MRE-2-13 complex (PDB code: ) and a series of mutants K51A, R84A and R87A, we applied molecular dynamics (MD) simulation and molecular mechanics generalized Born surface area (MM-GBSA) free energy calculations to examine the role of the tvMyb2 protein in recognition interaction. The simulation results indicate that tvMyb2 becomes stable when it binds the DNA duplex. A series of mutants, K51A, R84A and R87A, have been followed, and the results of statistical analyses of the H-bond and hydrophobic contacts show that some residues have significant influence on recognition and binding to ap65-1 DNA. Our work gives important information to understand the interactions of tvMyb2 with ap65-1. PMID:26548411

  2. AP-1-mediated chromatin looping regulates ZEB2 transcription: new insights into TNFα-induced epithelial-mesenchymal transition in triple-negative breast cancer.

    PubMed

    Qiao, Yichun; Shiue, Chiou-Nan; Zhu, Jian; Zhuang, Ting; Jonsson, Philip; Wright, Anthony P H; Zhao, Chunyan; Dahlman-Wright, Karin

    2015-04-10

    The molecular determinants of malignant cell behaviour in triple-negative breast cancer (TNBC) are poorly understood. Recent studies have shown that regulators of epithelial-mesenchymal transition (EMT) are potential therapeutic targets for TNBC. In this study, we demonstrate that the inflammatory cytokine TNFα induces EMT in TNBC cells via activation of AP-1 signaling and subsequently induces expression of the EMT regulator ZEB2. We also show that TNFα activates both the PI3K/Akt and MAPK/ERK pathways, which act upstream of AP-1. We further investigated in detail AP-1 regulation of ZEB2 expression. We show that two ZEB2 transcripts derived from distinct promoters are both expressed in breast cancer cell lines and breast tumor samples. Using the chromosome conformation capture assay, we demonstrate that AP-1, when activated by TNFα, binds to a site in promoter 1b of the ZEB2 gene where it regulates the expression of both promoter 1b and 1a, the latter via mediating long range chromatin interactions. Overall, this work provides a plausible mechanism for inflammation-induced metastatic potential in TNBC, involving a novel regulatory mechanism governing ZEB2 isoform expression. PMID:25762639

  3. Tankyrase inhibition aggravates kidney injury in the absence of CD2AP.

    PubMed

    Kuusela, S; Wang, H; Wasik, A A; Suleiman, H; Lehtonen, S

    2016-01-01

    Inappropriate activation of the Wnt/β-catenin pathway has been indicated in podocyte dysfunction and injury, and shown to contribute to the development and progression of nephropathy. Tankyrases, multifunctional poly(ADP-ribose) polymerase (PARP) superfamily members with features of both signaling and cytoskeletal proteins, antagonize Wnt/β-catenin signaling. We found that tankyrases interact with CD2-associated protein (CD2AP), a protein essential for kidney ultrafiltration as CD2AP-knockout (CD2AP-/-) mice die of kidney failure at the age of 6-7 weeks. We further observed that tankyrase-mediated total poly-(ADP-ribosyl)ation (PARylation), a post-translational modification implicated in kidney injury, was increased in mouse kidneys and cultured podocytes in the absence of CD2AP. The data revealed increased activity of β-catenin, and upregulation of lymphoid enhancer factor 1 (LEF1) (mediator of Wnt/β-catenin pathway) and fibronectin (downstream target of Wnt/β-catenin) in CD2AP-/- podocytes. Total PARylation and active β-catenin were reduced in CD2AP-/- podocytes by tankyrase inhibitor XAV939 treatment. However, instead of ameliorating podocyte injury, XAV939 further upregulated LEF1, failed to downregulate fibronectin and induced plasminogen activator inhibitor-1 (PAI-1) that associates with podocyte injury. In zebrafish, administration of XAV939 to CD2AP-depleted larvae aggravated kidney injury and increased mortality. Collectively, the data reveal sustained activation of the Wnt/β-catenin pathway in CD2AP-/- podocytes, contributing to podocyte injury. However, we observed that inhibition of the PARylation activity of tankyrases in the absence of CD2AP was deleterious to kidney function. This indicates that balance of the PARylation activity of tankyrases, maintained by CD2AP, is essential for normal kidney function. Furthermore, the data reveal that careful contemplation is required when targeting Wnt/β-catenin pathway to treat proteinuric kidney

  4. APS: 125 years of progress of physiology as a scientific discipline and a profession.

    PubMed

    Carroll, Robert G; Frank, Martin; Ra'anan, Alice; Matyas, Marsha L

    2013-03-01

    The Experimental Biology 2012 meeting in San Diego, CA, included events to celebrate the 125th anniversary of the founding of the American Physiological Society (APS) and reflect on the recent accomplishments of the society. Most of the APS activities in the past quarter century were guided by a series of strategic plans. Membership in the APS increased by 76% since 1987, and there are now 8,342 regular members of the society, including an expansion of international members to 26% of APS's membership. The numbers of elected officers and committee members have expanded to accommodate this larger membership. APS Publications changed dramatically during this time, having adopted online submission and review of manuscripts as well as a streamlined review and publications process that have significantly shortened the period from submission to acceptance to publication. Compared with 1987, the number of manuscripts submitted has increased by 80% and the number of printed pages increased by 52%. In addition to the refinement of the Experimental Biology meeting (Federation of American Societies for Experimental Biology meeting before 1993) format, APS launched a specialty conference program in the 1990s. The educational offerings of APS also dramatically expanded in the past 25 yr, often supported by external grants and contracts. APS education programs now support physiology education and science awareness at K-12, undergraduate, graduate, and professional levels as well as continuing education of its members. The founding of APS was tied to the need for effective advocacy, and APS continues to meet those goals through its Science Policy Committee and Animal Care and Experimentation Committees. At its 125th birthday, APS continues to serve the discipline and the needs of its membership. PMID:23471242

  5. Transient Fcho1/2⋅Eps15/R⋅AP-2 Nanoclusters Prime the AP-2 Clathrin Adaptor for Cargo Binding.

    PubMed

    Ma, Li; Umasankar, Perunthottathu K; Wrobel, Antoni G; Lymar, Anastasia; McCoy, Airlie J; Holkar, Sachin S; Jha, Anupma; Pradhan-Sundd, Tirthadipa; Watkins, Simon C; Owen, David J; Traub, Linton M

    2016-06-01

    Clathrin-coated vesicles form by rapid assembly of discrete coat constituents into a cargo-sorting lattice. How the sequential phases of coat construction are choreographed is unclear, but transient protein-protein interactions mediated by short interaction motifs are pivotal. We show that arrayed Asp-Pro-Phe (DPF) motifs within the early-arriving endocytic pioneers Eps15/R are differentially decoded by other endocytic pioneers Fcho1/2 and AP-2. The structure of an Eps15/R⋅Fcho1 μ-homology domain complex reveals a spacing-dependent DPF triad, bound in a mechanistically distinct way from the mode of single DPF binding to AP-2. Using cells lacking FCHO1/2 and with Eps15 sequestered from the plasma membrane, we establish that without these two endocytic pioneers, AP-2 assemblies are fleeting and endocytosis stalls. Thus, distinct DPF-based codes within the unstructured Eps15/R C terminus direct the assembly of temporary Fcho1/2⋅Eps15/R⋅AP-2 ternary complexes to facilitate conformational activation of AP-2 by the Fcho1/2 interdomain linker to promote AP-2 cargo engagement. PMID:27237791

  6. AP-1/σ1A and AP-1/σ1B adaptor-proteins differentially regulate neuronal early endosome maturation via the Rab5/Vps34-pathway

    PubMed Central

    Candiello, Ermes; Kratzke, Manuel; Wenzel, Dirk; Cassel, Dan; Schu, Peter

    2016-01-01

    The σ1 subunit of the AP-1 clathrin-coated-vesicle adaptor-protein complex is expressed as three isoforms. Tissues express σ1A and one of the σ1B and σ1C isoforms. Brain is the tissue with the highest σ1A and σ1B expression. σ1B-deficiency leads to severe mental retardation, accumulation of early endosomes in synapses and fewer synaptic vesicles, whose recycling is slowed down. AP-1/σ1A and AP-1/σ1B regulate maturation of these early endosomes into multivesicular body late endosomes, thereby controlling synaptic vesicle protein transport into a degradative pathway. σ1A binds ArfGAP1, and with higher affinity brain-specific ArfGAP1, which bind Rabex-5. AP-1/σ1A-ArfGAP1-Rabex-5 complex formation leads to more endosomal Rabex-5 and enhanced, Rab5GTP-stimulated Vps34 PI3-kinase activity, which is essential for multivesicular body endosome formation. Formation of AP-1/σ1A-ArfGAP1-Rabex-5 complexes is prevented by σ1B binding of Rabex-5 and the amount of endosomal Rabex-5 is reduced. AP-1 complexes differentially regulate endosome maturation and coordinate protein recycling and degradation, revealing a novel molecular mechanism by which they regulate protein transport besides their established function in clathrin-coated-vesicle formation. PMID:27411398

  7. APS Alternative Fuel (Hydrogen) Pilot Plant - Monitoring System Report

    SciTech Connect

    James Francfort; Dimitri Hochard

    2005-07-01

    The U.S. Department of Energy’s (DOE’s) Advanced Vehicle Testing Activity (AVTA), along with Electric Transportation Applications and Arizona Pubic Service (APS), is monitoring the operations of the APS Alternative Fuel (Hydrogen) Pilot Plant to determine the costs to produce hydrogen fuels (including 100% hydrogen as well as hydrogen and compressed natural gas blends) for use by fleets and other operators of advanced-technology vehicles. The hydrogen fuel cost data will be used as benchmark data by technology modelers as well as research and development programs. The Pilot Plant can produce up to 18 kilograms (kg) of hydrogen per day by electrolysis. It can store up to 155 kg of hydrogen at various pressures up to 6,000 psi. The dispenser island can fuel vehicles with 100% hydrogen at 5,000 psi and with blends of hydrogen and compressed natural gas at 3,600 psi. The monitoring system was designed to track hydrogen delivery to each of the three storage areas and to monitor the use of electricity on all major equipment in the Pilot Plant, including the fuel dispenser island. In addition, water used for the electrolysis process is monitored to allow calculation of the total cost of plant operations and plant efficiencies. The monitoring system at the Pilot Plant will include about 100 sensors when complete (50 are installed to date), allowing for analysis of component, subsystems, and plant-level costs. The monitoring software is mostly off-the-shelve, with a custom interface. The majority of the sensors input to the Programmable Automation Controller as 4- to 20-mA analog signals. The plant can be monitored over of the Internet, but the control functions are restricted to the control room equipment. Using the APS general service plan E32 electric rate of 2.105 cents per kWh, during a recent eight-month period when 1,200 kg of hydrogen was produced and the plant capacity factor was 26%, the electricity cost to produce one kg of hydrogen was $3.43. However, the

  8. Preface: 18th Aps-Sccm and 24th Airapt

    NASA Astrophysics Data System (ADS)

    Collins, Gilbert; Moore, David S.; Yoo, Choong-Shik; Buttler, William; Furlanetto, Michael; Evans, William

    2014-05-01

    The 18th Biennial International Conference of the APS Topical Group on Shock Compression of Condensed Matter in conjunction with the 24th Biennial International Conference of the International Association for the Advancement of High Pressure Science & Technology (AIRAPT) was held at the Westin Hotel in Seattle, Washington from 7-12 July, 2013. This is only the second time that these two organizations have held a Joint Conference — the first was 20 years previous (1993) in Colorado Springs, Colorado. Seattle was chosen for this joint conference because of its central location for the world-wide attendees as well as its metropolitan vibrancy. The scientific program consisted of 858 scheduled presentations organized into 23 topical areas and included contributed (537), invited (95), and plenary (6) lectures, as well as two poster sessions with 110 posters each. The scientific focus of the Joint Conference was on fundamental and applied research topics related to the static or dynamic compression of condensed matter. This multidisciplinary field of research encompasses areas of physics, chemistry, materials science, mechanics, geophysics and planetary physics, and applied mathematics. Experimental, computational and theoretical studies all play important roles. The organizers endeavored to intertwine static and dynamic experimental alongside computational and theoretical studies of similar materials in the organization of the sessions. This goal was aided by the addition of three special focus sessions on deep carbon budget, high energy density materials, and dynamic response of materials. 722 scientists and engineers from 25 countries registered at the conference, including 132 students from 12 countries. The attendee countries represented included: Argentina (2), Australia (2), Brazil (3), Canada (25), China (22), Czech Republic (2), France (35), Germany (19), India (6), Israel (21), Italy (10), Japan (49), Netherlands (1), Poland (1), Portugal (2), Russia (26

  9. AP-2α downregulation by cigarette smoke condensate is counteracted by p53 in human lung cancer cells.

    PubMed

    Meng, Xiangjun; Meng, Cuida; Yang, Bing; Zhao, Li; Sun, Xuefei; Su, Yun; Liu, Hongyang; Fan, Feiyue; Liu, Xiaodong; Jia, Lili

    2014-10-01

    Cumulative findings have demonstrated that the dysregulation of tumor suppressor genes may be implicated in cigarette smoke-induced carcinogenesis. Activating enhancer-binding protein 2 (AP-2) is a eukaryotic transcriptional factor that plays a significant role in embryonic development and tumorigenesis. The vertebrate AP-2 family consists of AP-2α, AP-2β, AP-2γ, AP-2δ and AP-2ε. Previous studies have suggested that cigarette smoking disrupts AP-2 regulation. In the present study, we investigated the effects of cigarette smoke condensate (CSC) on AP-2α expression in human lung cancer cell lines (NCI-H1299, NCI-H446 and A549), as well as the potential mechanisms involved. Using RT-qPCR, we found that CSC decreased AP-2α expression by suppressing its transcription in human lung cancer cell lines, particularly in p53-deficient NCI-H1299 cells. Western blotting and luciferase assays were implemented and we found that the restoration of p53 expression rescued the NCI-H1299 cells from CSC-induced AP-2α loss, while the silencing of p53 resulted in increased AP-2α loss induced by CSC, suggesting an antagonizing role of p53 in the regulation of AP-2α by CSC. Our results indicate that AP-2α downregulation may be involved in smoke-induced lung carcinogenesis. PMID:25050743

  10. Neuronal adaptation involves rapid expansion of the action potential initiation site.

    PubMed

    Scott, Ricardo S; Henneberger, Christian; Padmashri, Ragunathan; Anders, Stefanie; Jensen, Thomas P; Rusakov, Dmitri A

    2014-01-01

    Action potential (AP) generation is the key to information-processing in the brain. Although APs are normally initiated in the axonal initial segment, developmental adaptation or prolonged network activity may alter the initiation site geometry thus affecting cell excitability. Here we find that hippocampal dentate granule cells adapt their spiking threshold to the kinetics of the ongoing dendrosomatic excitatory input by expanding the AP-initiation area away from the soma while also decelerating local axonal spikes. Dual-patch soma-axon recordings combined with axonal Na(+) and Ca(2+) imaging and biophysical modelling show that the underlying mechanism involves distance-dependent inactivation of axonal Na(+) channels due to somatic depolarization propagating into the axon. Thus, the ensuing changes in the AP-initiation zone and local AP propagation could provide activity-dependent control of cell excitability and spiking on a relatively rapid timescale. PMID:24851940

  11. Neuronal adaptation involves rapid expansion of the action potential initiation site

    PubMed Central

    Scott, Ricardo S.; Henneberger, Christian; Padmashri, Ragunathan; Anders, Stefanie; Jensen, Thomas P.; Rusakov, Dmitri A.

    2014-01-01

    Action potential (AP) generation is the key to information-processing in the brain. Although APs are normally initiated in the axonal initial segment, developmental adaptation or prolonged network activity may alter the initiation site geometry thus affecting cell excitability. Here we find that hippocampal dentate granule cells adapt their spiking threshold to the kinetics of the ongoing dendrosomatic excitatory input by expanding the AP-initiation area away from the soma while also decelerating local axonal spikes. Dual-patch soma–axon recordings combined with axonal Na+ and Ca2+ imaging and biophysical modelling show that the underlying mechanism involves distance-dependent inactivation of axonal Na+ channels due to somatic depolarization propagating into the axon. Thus, the ensuing changes in the AP-initiation zone and local AP propagation could provide activity-dependent control of cell excitability and spiking on a relatively rapid timescale. PMID:24851940

  12. 10 CFR Appendix C to Part 52 - Design Certification Rule for the AP600 Design

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 10 Energy 2 2010-01-01 2010-01-01 false Design Certification Rule for the AP600 Design C Appendix C to Part 52 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSES, CERTIFICATIONS, AND APPROVALS FOR NUCLEAR POWER PLANTS Pt. 52, App. C Appendix C to Part 52—Design Certification Rule for the AP600 Design I. Introduction Appendix C...

  13. 10 CFR Appendix C to Part 52 - Design Certification Rule for the AP600 Design

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 10 Energy 2 2014-01-01 2014-01-01 false Design Certification Rule for the AP600 Design C Appendix C to Part 52 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSES, CERTIFICATIONS, AND APPROVALS FOR NUCLEAR POWER PLANTS Pt. 52, App. C Appendix C to Part 52—Design Certification Rule for the AP600 Design I. Introduction Appendix C...

  14. Coming Soon: CADRE (Career Advancement and Development Resources and Education) website for all APS members

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Council of the American Phytopathological Society (APS) approved an initiative in February 2013 to create a web resource called CADRE (Career Advancement and Development Resources and Education). CADRE is to provide APS members an archive of articles, videos, and webinars about a variety of prof...

  15. The Freshman Nine: Helping High School Freshmen Be Successful in AP Human Geography

    ERIC Educational Resources Information Center

    Garner, Jennifer

    2012-01-01

    Teaching AP Human Geography to freshmen seems like a daunting task and while there are many arguments both for and against offering the course to freshmen, for many teachers it is reality. In this article, the author offers nine tips to help high school freshmen be successful in the course and on the AP exam.

  16. Development of cotton gin PM2.5 emission factors for EPA’S AP-42

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Compilation of Air Pollution Emission Factors (AP-42) emission factors are assigned ratings, from A (Excellent) to E (Poor), based on the quality of data used to develop them. AP-42 currently contains no PM2.5 cotton gin emission factors. In an effort to develop science-based data for regulating...

  17. AP American History and the History Major: Keeping Body and Soul Together.

    ERIC Educational Resources Information Center

    Holbo, Paul S.

    For college-level American History, in the high school advanced placement (AP) program and on university campuses, these are the best of times and the worst of times. For the American History AP program, the early 1970's were difficult times, with the examinations under attack as elitist and irrelevant to contemporary problems. The program…

  18. Environmental and Sustainable Technology Evaluation: Mold-Resistant Armacell Insulation--Armacell LLC, AP Armaflex Black

    EPA Science Inventory

    The ESTE test program measured the mold resistance of Armacell AP Armaflex Black insulation. Tests for emissions of VOCs and formaldehyde were also performed. AP Armaflex Roll Insulation is a black flexible closed-cell, fiber-free elastomeric thermal insulation. The expanded clos...

  19. Foundations for College and Beyond: Looking Back on AP Art History

    ERIC Educational Resources Information Center

    Schoenbohm, Laurel

    2013-01-01

    It was years after this author's AP Art History course in high school, and two years after college. She and some friends decided to fill a day during the Thanksgiving visits appreciating fine art. Prior to that AP course her senior year of high school, touring an art museum had seemed like the equivalent of going to the dentist. But after…

  20. Amphioxus and lamprey AP-2 genes: implications for neural crest evolution and migration patterns

    NASA Technical Reports Server (NTRS)

    Meulemans, Daniel; Bronner-Fraser, Marianne

    2002-01-01

    The neural crest is a uniquely vertebrate cell type present in the most basal vertebrates, but not in cephalochordates. We have studied differences in regulation of the neural crest marker AP-2 across two evolutionary transitions: invertebrate to vertebrate, and agnathan to gnathostome. Isolation and comparison of amphioxus, lamprey and axolotl AP-2 reveals its extensive expansion in the vertebrate dorsal neural tube and pharyngeal arches, implying co-option of AP-2 genes by neural crest cells early in vertebrate evolution. Expression in non-neural ectoderm is a conserved feature in amphioxus and vertebrates, suggesting an ancient role for AP-2 genes in this tissue. There is also common expression in subsets of ventrolateral neurons in the anterior neural tube, consistent with a primitive role in brain development. Comparison of AP-2 expression in axolotl and lamprey suggests an elaboration of cranial neural crest patterning in gnathostomes. However, migration of AP-2-expressing neural crest cells medial to the pharyngeal arch mesoderm appears to be a primitive feature retained in all vertebrates. Because AP-2 has essential roles in cranial neural crest differentiation and proliferation, the co-option of AP-2 by neural crest cells in the vertebrate lineage was a potentially crucial event in vertebrate evolution.

  1. 75 FR 20774 - Establishment of Class E Airspace; Fort A.P. Hill, VA

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-21

    ... final rule with a request for comments in the Federal Register on December 7, 2009 (74 FR 63974), Docket... Federal Aviation Administration 14 CFR Part 71 Establishment of Class E Airspace; Fort A.P. Hill, VA... Register December 7, 2009 that establishes Class E airspace at Fort A.P. Hill, VA. DATES: Effective...

  2. 77 FR 35308 - Proposed Amendment of Restricted Area R-6601; Fort A.P. Hill, VA

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-06-13

    .... 10854, 24 FR 9565, 3 CFR, 1959-1963 Comp., p. 389. Sec. 73.66 (Amended) 2. Sec. 73.66 is amended as... Federal Aviation Administration 14 CFR Part 73 Proposed Amendment of Restricted Area R-6601; Fort A.P... restricted area R-6601, Fort A.P. Hill, VA. The U. S. Army requested this action to provide the...

  3. APS: 125 Years of Progress of Physiology as a Scientific Discipline and a Profession

    ERIC Educational Resources Information Center

    Carroll, Robert G.; Frank, Martin; Ra'anan, Alice; Matyas, Marsha L.

    2013-01-01

    The Experimental Biology 2012 meeting in San Diego, CA, included events to celebrate the 125th anniversary of the founding of the American Physiological Society (APS) and reflect on the recent accomplishments of the society. Most of the APS activities in the past quarter century were guided by a series of strategic plans. Membership in the APS…

  4. Ten Things to Consider When Teaching AP U.S. History

    ERIC Educational Resources Information Center

    Libresco, Andrea S.

    2013-01-01

    This article describes 10 recommendations for creativity, higher-order thinking, and meaningful learning activities that can be used to guide teachers in constructing an engaging AP course: (1) Be on the committee that decides how students will be selected for AP; (2) Maximize time and connections through blocks of time with an English colleague;…

  5. Understanding and Using the New Guided-Inquiry AP Chemistry Laboratory Manual

    ERIC Educational Resources Information Center

    Cacciatore, Kristen L.

    2014-01-01

    To support teaching and learning in the advanced placement (AP) chemistry laboratory, the College Board published a laboratory manual, "AP Chemistry Guided-Inquiry Experiments: Applying the Science Practices," in 2013 as part of the redesigned course. This article provides a discussion of the rationale for the existence of the manual as…

  6. AP180-mediated trafficking of Vamp7B limits homotypic fusion of Dictyostelium contractile vacuoles.

    PubMed

    Wen, Yujia; Stavrou, Irene; Bersuker, Kirill; Brady, Rebecca J; De Lozanne, Arturo; O'Halloran, Theresa J

    2009-10-01

    Clathrin-coated vesicles play an established role in endocytosis from the plasma membrane, but they are also found on internal organelles. We examined the composition of clathrin-coated vesicles on an internal organelle responsible for osmoregulation, the Dictyostelium discoideum contractile vacuole. Clathrin puncta on contractile vacuoles contained multiple accessory proteins typical of plasma membrane-coated pits, including AP2, AP180, and epsin, but not Hip1r. To examine how these clathrin accessory proteins influenced the contractile vacuole, we generated cell lines that carried single and double gene knockouts in the same genetic background. Single or double mutants that lacked AP180 or AP2 exhibited abnormally large contractile vacuoles. The enlarged contractile vacuoles in AP180-null mutants formed because of excessive homotypic fusion among contractile vacuoles. The SNARE protein Vamp7B was mislocalized and enriched on the contractile vacuoles of AP180-null mutants. In vitro assays revealed that AP180 interacted with the cytoplasmic domain of Vamp7B. We propose that AP180 directs Vamp7B into clathrin-coated vesicles on contractile vacuoles, creating an efficient mechanism for regulating the internal distribution of fusion-competent SNARE proteins and limiting homotypic fusions among contractile vacuoles. Dictyostelium contractile vacuoles offer a valuable system to study clathrin-coated vesicles on internal organelles within eukaryotic cells. PMID:19692567

  7. Advanced Placement Strategy: A Framework for Identifying School-Level Barriers to AP Success. Policy Brief

    ERIC Educational Resources Information Center

    Batiwalla, Mary

    2014-01-01

    In 2013, Tennessee counted nearly 7,000 students in the senior cohort whose academic skills when they entered high school suggested they were on track to earn college credits through Advanced Placement (AP) exams. Yet just over half of these students actually graduated with an AP credit, and less than a third of the economically disadvantaged…

  8. The Rush to Take More AP Courses Hurts Students, High Schools, and Colleges

    ERIC Educational Resources Information Center

    Oxtoby, David W.

    2007-01-01

    In most schools, the rush toward Advanced Placement (AP) courses goes on unabated. In 2006 the number of students taking AP exams increased almost 10 percent over the preceding year. This is largely because of the growing intensity of the admissions game and the urge on the part of college applicants to seek out every advantage. In applying to…

  9. The Rush to Take More AP Courses Hurts Students, High Schools, and Colleges

    ERIC Educational Resources Information Center

    Oxtoby, David W.

    2007-01-01

    In most schools, the rush toward Advanced Placement (AP) courses goes on unabated. In 2006 the number of students taking AP exams increased almost 10 percent over the preceding year, according to the College Board. This is largely because of the growing intensity of the admissions game and the urge on the part of college applicants to seek out…

  10. AP-1 family members act with Sox9 to promote chondrocyte hypertrophy.

    PubMed

    He, Xinjun; Ohba, Shinsuke; Hojo, Hironori; McMahon, Andrew P

    2016-08-15

    An analysis of Sox9 binding profiles in developing chondrocytes identified marked enrichment of an AP-1-like motif. Here, we have explored the functional interplay between Sox9 and AP-1 in mammalian chondrocyte development. Among AP-1 family members, Jun and Fosl2 were highly expressed within prehypertrophic and early hypertrophic chondrocytes. Chromatin immunoprecipitation followed by DNA sequencing (ChIP-seq) showed a striking overlap in Jun- and Sox9-bound regions throughout the chondrocyte genome, reflecting direct binding of each factor to the same enhancers and a potential for protein-protein interactions within AP-1- and Sox9-containing complexes. In vitro reporter analysis indicated that direct co-binding of Sox9 and AP-1 at target motifs promoted gene activity. By contrast, where only one factor can engage its DNA target, the presence of the other factor suppresses target activation consistent with protein-protein interactions attenuating transcription. Analysis of prehypertrophic chondrocyte removal of Sox9 confirmed the requirement of Sox9 for hypertrophic chondrocyte development, and in vitro and ex vivo analyses showed that AP-1 promotes chondrocyte hypertrophy. Sox9 and Jun co-bound and co-activated a Col10a1 enhancer in Sox9 and AP-1 motif-dependent manners consistent with their combined action promoting hypertrophic gene expression. Together, the data support a model in which AP-1 family members contribute to Sox9 action in the transition of chondrocytes to the hypertrophic program. PMID:27471255

  11. 77 FR 8848 - Application for New Awards; Advanced Placement (AP) Test Fee Program

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-15

    ... Application for New Awards; Advanced Placement (AP) Test Fee Program AGENCY: Office of Elementary and Secondary Education, Department of Education. ACTION: Notice. Overview Information: Advanced Placement Test.... Full Text of Announcement I. Funding Opportunity Description Purpose of Program: The AP Test...

  12. 78 FR 19691 - Applications for New Awards; Advanced Placement (AP) Test Fee Program

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-02

    ... Applications for New Awards; Advanced Placement (AP) Test Fee Program AGENCY: Office of Elementary and Secondary Education, Department of Education. ACTION: Notice. Overview Information Advanced Placement Test... Announcement I. Funding Opportunity Description Purpose of Program: The AP Test Fee program awards grants...

  13. PSAT Component Scores as a Predictor of Success on AP Exam Performance for Diverse Students

    ERIC Educational Resources Information Center

    Richardson, Cristianne C.; Gonzalez, Alejandro; Leal, Lonnie; Castillo, Mary Z.; Carman, Carol A.

    2016-01-01

    While studies have shown the positive effect of the Advanced Placement (AP) program on college readiness, there are still barriers preventing minority and low socioeconomic status (SES) students who possess high academic potential from participating in the opportunity that AP courses offer. One tool that could help identify students for…

  14. AP-42 ADDITIONS AND REVISIONS - GRAIN LOADING ONTO SHIPS AND BARGES

    EPA Science Inventory

    This project develops emission factors, etc., for the grain shipping industry which are appended to AP-42. AP42 is a massive collection of material which describes processes which generate air emissions and presents emission factors and control effectiveness information. As reso...

  15. Social Effects of Hispanic Students Enrolled in an AP Class in Middle School

    ERIC Educational Resources Information Center

    Shiu, Alexandra; Kettler, Todd; Johnsen, Susan K.

    2009-01-01

    This article describes the effects of placing Spanish-speaking students in an Advanced Placement (AP) Spanish Language course in the 8th grade. Fifty-eight Hispanic students in the AP class and a similar comparison group answered survey questions on parental involvement, peers, sense of belonging at school, and academic aspirations. Data gathered…

  16. The History of the APS Shock Compression of Condensed Matter Topical Group

    SciTech Connect

    Forbes, J W

    2001-05-02

    In order to provide broader scientific recognition and to advance the science of shock compressed condensed matter, a group of American Physical Society (APS) members worked within the Society to make this field an active part of the APS. Individual papers were presented at APS meetings starting in the 1940's and shock wave sessions were organized starting with the 1967 Pasadena meeting. Shock wave topical conferences began in 1979 in Pullman, WA. Signatures were obtained on a petition in 1984 from a balanced cross-section of the shock wave community to form an APS Topical Group (TG). The APS Council officially accepted the formation of the Shock Compression of Condensed Matter (SCCM) TG at its October 1984 meeting. This action firmly aligned the shock wave field with a major physical science organization. Most early topical conferences were sanctioned by the APS while those held after 1992 were official APS meetings. The topical group organizes a shock wave topical conference in odd numbered years while participating in shock wave/high pressure sessions at APS general meetings in even numbered years.

  17. History of the APS Topical Group on Shock Compression of Condensed Matter

    SciTech Connect

    Forbes, J W

    2001-10-19

    In order to provide broader scientific recognition and to advance the science of shock compressed condensed matter, a group of American Physical Society (APS) members worked within the Society to make this field an active part of the APS. Individual papers were presented at APS meetings starting in the 1940's and shock wave sessions were organized starting with the 1967 Pasadena meeting. Shock wave topical conferences began in 1979 in Pullman, WA. Signatures were obtained on a petition in 1984 from a balanced cross-section of the shock wave community to form an APS Topical Group (TG). The APS Council officially accepted the formation of the Shock Compression of Condensed Matter (SCCM) TG at its October 1984 meeting. This action firmly aligned the shock wave field with a major physical science organization. Most early topical conferences were sanctioned by the APS while those held after 1992 were official APS meetings. The topical group organizes a shock wave topical conference in odd numbered years while participating in shock wavehigh pressure sessions at APS general meetings in even numbered years.

  18. WCPSS Advanced Placement (AP) Results, 2010-11. D&A Report No. 11.25

    ERIC Educational Resources Information Center

    Gilleland, Kevin

    2012-01-01

    One method of delivering college-level coursework to high school students is through the Advanced Placement (AP) program. Many colleges and universities provide credit to students who earn a qualifying score on any of the 34 available AP exams offered by the College Board. All qualified comprehensive high schools in the Wake County Public School…

  19. Updating AP Potential™ Expectancy Tables Involving PSAT/NMSQT® Writing. Research Notes. RN-35

    ERIC Educational Resources Information Center

    Ewing, Maureen; Camara, Wayne J.; Millsap, Roger E.; Milewski, Glenn B.

    2007-01-01

    AP Potential™ is a data-driven tool offered by the College Board that uses scores from the PSAT/NMSQT® to identify students who have the potential to succeed in Advanced Placement Program® (AP®) courses (College Board, 2007). Research showing a moderate-to-strong correlation between PSAT/NMSQT scores and AP Exam scores serves as the basis for this…

  20. Glutathione peroxidase-1 inhibits UVA-induced AP-2{alpha} expression in human keratinocytes

    SciTech Connect

    Yu Lei; Venkataraman, Sujatha; Coleman, Mitchell C.; Spitz, Douglas R.; Wertz, Philip W.; Domann, Frederick E. . E-mail: frederick-domann@uiowa.edu

    2006-12-29

    In this study, we found a role for H{sub 2}O{sub 2} in UVA-induced AP-2{alpha} expression in the HaCaT human keratinocyte cell line. UVA irradiation not only increased AP-2{alpha}, but also caused accumulation of H{sub 2}O{sub 2} in the cell culture media, and H{sub 2}O{sub 2} by itself could induce the expression of AP-2{alpha}. By catalyzing the removal of H{sub 2}O{sub 2} from cells through over-expression of GPx-1, induction of AP-2{alpha} expression by UVA was abolished. Induction of transcription factor AP-2{alpha} by UVA had been previously shown to be mediated through the second messenger ceramide. We found that not only UVA irradiation, but also H{sub 2}O{sub 2} by itself caused increases of ceramide in HaCaT cells, and C2-ceramide added to cells induced the AP-2{alpha} signaling pathway. Finally, forced expression of GPx-1 eliminated UVA-induced ceramide accumulation as well as AP-2{alpha} expression. Taken together, these findings suggest that GPx-1 inhibits UVA-induced AP-2{alpha} expression by suppressing the accumulation of H{sub 2}O{sub 2}.

  1. AP-42 ADDITIONS AND REVISIONS - INTEGRATED IRON AND STEEL INDUSTRY - STEEL MINI MILLS

    EPA Science Inventory

    This project develops emission factors, etc., for the integrated iron and steel industry which are incorporated into AP-42. AP-42 is a massive collection of information concerning processes which generate air emissions and presents emission factors and control effectiveness infor...

  2. Growth and Achievement Trends of Advanced Placement (AP) Exams in American High Schools

    ERIC Educational Resources Information Center

    Judson, Eugene; Hobson, Angela

    2015-01-01

    This exploratory study examined and compared overall trends in growth and student achievement of the Advanced Placement (AP) program. Using data from the past two decades, analyses indicated there has been steady and extensive growth of AP participation, particularly among underclassmen and some minority groups. However, overall achievement, as…

  3. The 8th Annual AP[R] Report to the Nation

    ERIC Educational Resources Information Center

    College Board, 2012

    2012-01-01

    In classrooms around the country, AP (Advanced Placement) teachers are preparing students for tomorrow by teaching them how to think and learn today. AP students learn to construct solid arguments, test theories, and see many sides of an issue--the kind of thinking that solves tough problems both in and outside the classroom, in college and…

  4. The 6th Annual AP[R] Report to the Nation

    ERIC Educational Resources Information Center

    College Board, 2010

    2010-01-01

    Educators across the United States continue to enable a wider and more ethnically diverse proportion of students to achieve success in AP[R]. Significant inequities remain, however, which can result in traditionally underserved students not receiving the type of AP (Advanced Placement) opportunities that can best prepare them for college success.…

  5. Involvement of the AP-1 Adaptor Complex in Early Steps of Phagocytosis and Macropinocytosis

    PubMed Central

    Lefkir, Yaya; Malbouyres, Marilyne; Gotthardt, Daniel; Ozinsky, Adrian; Cornillon, Sophie; Bruckert, Franz; Aderem, Alan A.; Soldati, Thierry; Cosson, Pierre; Letourneur, François

    2004-01-01

    The best described function of the adaptor complex-1 (AP-1) is to participate in the budding of clathrin-coated vesicles from the trans-Golgi network and endosomes. Here, we show that AP-1 is also localized to phagocytic cups in murine macrophages as well as in Dictyostelium amoebae. AP-1 is recruited to phagosomal membranes at this early stage of phagosome formation and rapidly dissociates from maturing phagosomes. To establish the role of AP-1 in phagocytosis, we made used of Dictyostelium mutant cells (apm1- cells) disrupted for AP-1 medium chain. In this mutant, phagocytosis drops by 60%, indicating that AP-1 is necessary for efficient phagocytosis. Furthermore, phagocytosis in apm1- cells is more affected for large rather than small particles, and cells exhibiting incomplete engulfment are then often observed. This suggests that AP-1 could participate in the extension of the phagocytic cup. Interestingly, macropinocytosis, a process dedicated to fluid-phase endocytosis and related to phagocytosis, is also impaired in apm1- cells. In summary, our data suggest a new role of AP-1 at an early stage of phagosome and macropinosome formation. PMID:14617812

  6. Back to the Future: Merit or Equity in AP Social Studies?

    ERIC Educational Resources Information Center

    Stevens, Robert

    2013-01-01

    In an effort to address severe budget deficits at both the state and local levels, schools and educational programs are being asked to trim budgets. The Advanced Placement Program is one program that will certainly be scrutinized. This article presents a general overview of AP social studies, a brief history of the AP social studies program, and…

  7. Replication bypass and mutagenic effect of alpha-deoxyadenosine site-specifically incorporated into single-stranded vectors.

    PubMed Central

    Shimizu, H; Yagi, R; Kimura, Y; Makino, K; Terato, H; Ohyama, Y; Ide, H

    1997-01-01

    alpha-2'-Deoxyadenosine (alpha) is a major adenine lesion produced by gamma-ray irradiation of DNA under anoxic conditions. In this study, single-stranded recombinant M13 vectors containing alpha were constructed and transfected into Escherichia coli to assess lethal and mutagenic effects of this lesion. The data for alpha were further compared with those obtained with M13 vectors containing normal A or a model abasic site (F) at the same site. The transfection assay revealed that alpha constituted a moderate block to DNA replication. The in vivo replication capacity to pass through alpha was approximately 20% relative to normal A, but 20-fold higher than that of F constituting an almost absolute replication block. Similar data were obtained by in vitro replication of oligonucleotide templates containing alpha or F by E.coli DNA polymerase I. The mutagenic consequence of replicating M13 DNA containing alpha was analyzed by direct DNA sequencing of progeny phage. Mutagenesis was totally targeted at the site of alpha introduced into the vector. Mutation was exclusively a single nucleotide deletion and no base substitutions were detected. The deletion frequency associated alpha was dependent on the 3'-nearest neighbor base: with the 3'-nearest neighbor base T mutation (deletion) frequency was 26%, whereas 1% with the 3'-nearest neighbor base G. A possible mechanism of the single nucleotide deletion associated with alpha is discussed on the basis of the misinsertion-strand slippage model. PMID:9016601

  8. Human kidney anion exchanger 1 interacts with adaptor-related protein complex 1 μ1A (AP-1 mu1A).

    PubMed

    Sawasdee, Nunghathai; Junking, Mutita; Ngaojanlar, Piengpaga; Sukomon, Nattakan; Ungsupravate, Duangporn; Limjindaporn, Thawornchai; Akkarapatumwong, Varaporn; Noisakran, Sansanee; Yenchitsomanus, Pa-Thai

    2010-10-01

    Kidney anion exchanger 1 (kAE1) mediates chloride (Cl⁻) and bicarbonate (HCO₃⁻) exchange at the basolateral membrane of kidney α-intercalated cells. Impaired trafficking of kAE1 leads to defect of the Cl⁻/HCO₃⁻ exchange at the basolateral membrane and failure of proton (H+) secretion at the apical membrane, causing a kidney disease--distal renal tubular acidosis (dRTA). To gain a better insight into kAE1 trafficking, we searched for proteins physically interacting with the C-terminal region of kAE1 (Ct-kAE1), which contains motifs crucial for intracellular trafficking, by a yeast two-hybrid (Y2H) system. An adaptor-related protein complex 1 μ1A (AP-1 mu1A) subunit was found to interact with Ct-kAE1. The interaction between either Ct-kAE1 or full-length kAE1 and AP-1 mu1A were confirmed in human embryonic kidney (HEK) 293T by co-immunoprecipitation, affinity co-purification, co-localization, yellow fluorescent protein (YFP)-based protein fragment complementation assay (PCA) and GST pull-down assay. The interacting site for AP-1 mu1A on Ct-kAE1 was found to be Y904DEV907, a subset of YXXØ motif. Interestingly, suppression of endogenous AP-1 mu1A in HEK 293T by small interfering RNA (siRNA) decreased membrane localization of kAE1 and increased its intracellular accumulation, suggesting for the first time that AP-1 mu1A is involved in the kAE1 trafficking of kidney α-intercalated cells. PMID:20833140

  9. SITE RANK

    EPA Science Inventory

    Site rank is formulated for ranking the relative hazard of contamination sources and vulnerability of drinking water wells. Site rank can be used with a variety of groundwater flow and transport models.

  10. Glossary for the implementation of Health in All Policies (HiAP).

    PubMed

    Freiler, Alix; Muntaner, Carles; Shankardass, Ketan; Mah, Catherine L; Molnar, Agnes; Renahy, Emilie; O'Campo, Patricia

    2013-12-01

    Health in All Policies (HiAP) is becoming increasingly popular as a governmental strategy to improve population health by coordinating action across health and non-health sectors. A variety of intersectoral initiatives may be used in HiAP that frame health determinants as the bridge between policies and health outcomes. The purpose of this glossary is to present concepts and terms useful in understanding the implementation of HiAP as a cross-sectoral policy. The concepts presented here were applied and elaborated over the course of case studies of HiAP in multiple jurisdictions, which used key informant interviews and the systematic review of literature to study the implementation of specific HiAP initiatives. PMID:23986493

  11. Copernicus observations of the Ap star Epsilon Ursae Majoris

    NASA Technical Reports Server (NTRS)

    Mallama, A. D.; Molnar, M. R.

    1977-01-01

    Spectral scans of the Ap star Epsilon UMa made with the Copernicus satellite show strong line blanketing from profuse Cr II and Fe II lines. In the spectral region covering 1900 to 3000 A, about 500 lines are present which suppress the apparent continuum by at least 15-30%. An accurate line-identification list is compiled showing Eu II present in addition to Mn II and Ni II. The identification of Eu II, however, rests on very stringent identification limits for Fe II. If these are relaxed, the existence of Eu II is dubious. There are no broad features in this spectral region which would suggest strong photoionization discontinuities by metals, but one feature near 2137 A might contain the photoionization edge due to Cr I 5S lying 0.94 eV above the ground level. However, a significant correlation between the line-blanketing strength and the amplitude of the OAO-2 ultraviolet light curves was found such that both monotonically increase in the same proportion toward shorter wavelengths. This gives additional strength to the suggestion that variations in the metal line-blanketing cause the observed photometric variations.

  12. Tandem Differential Mobility Analyzer/Aerodynamic Particle Sizer (APS) Handbook

    SciTech Connect

    Collins, D

    2010-06-18

    The tandem differential mobility analyzer (TDMA) is a single instrument that cycles through a series of complementary measurements of the physical properties of size-resolved submicron particles. In 2008, the TDMA was augmented through the addition of an aerodynamic particle sizer (APS), which extends the upper limit of the measured size distribution into the supermicron range. These two instruments are operated in parallel, but because they are controlled by a common computer and because the size distributions measured by the two are integrated in the produced datastreams, they are described together here. Throughout the day, the TDMA sequentially measures submicron aerosol size distributions and size-resolved hygroscopic growth distributions. More specifically, the instrument is operated as a scanning DMA to measure size distributions and as a TDMA to measure size-resolved hygroscopicity. A typical measurement sequence requires roughly 45 minutes. Each morning additional measurements are made of the relative humidity (RH) dependent hygroscopicity and temperature-dependent volatility of size-resolved particles. When the outside temperature and RH are within acceptable ranges, the hydration state of size-resolved particles is also characterized. The measured aerosol distributions complement the array of aerosol instruments in the Aerosol Observing System (AOS) and provide additional details of the light-scattering and cloud-nucleating characteristics of the aerosol.

  13. Arbitrary function generator for APS injector synchrotron correction magnets

    SciTech Connect

    Despe, O.D.

    1990-11-07

    The APS injector synchrotron ring measures about 368 m in circumference. In order to obtain the precision of the magnetic field required for the positron acceleration from 450 Mev to 7.7 Gev with low beam loss, eighty correction magnets are distributed around its circumference. These magnets provide the vernier field changes required for beam orbit correction during the acceleration phase of the injector synchrotron cycle. Because of mechanical imperfections in the construction, as well as installation of real dipole and multi-pole magnets, the exact field correction required at each correction magnet location is not known until a beam is actually accelerated. It is therefore essential that a means is provided to generate a correction field that is a function of the beam energy from injection until extraction for each correction magnet. The fairly large number of correction magnets in the system requires that the arbitrary function generator design be as simple as possible yet provide the required performance. An important, required performance feature is that the function can be changed or modified ``on the fly``, to provide the operator with a real-time feel during the tune up process. The arbitrary function generator described in this report satisfies these requirements.

  14. Trim69 regulates zebrafish brain development by ap-1 pathway

    PubMed Central

    Han, Ruiqin; Wang, Renxian; Zhao, Qing; Han, Yongqing; Zong, Shudong; Miao, Shiying; Song, Wei; Wang, Linfang

    2016-01-01

    Proteins belonging to the TRIM family have been implicated in a variety of cellular processes such as apoptosis, differentiation, neurogenesis, muscular physiology and innate immune responses. Trim69, previously identified as a novel gene cloned from a human testis cDNA library, has a homologous gene in zebrafish and this study focused on investigating the function of trim69 in zebrafish neurogenesis. Trim69 was found to be expressed in zebrafish embryo brain at the early stages. Knockdown of trim69 led to deformed brain development, obvious signs of apoptosis present in the head, and decreased expression of neuronal differentiation and stem cell markers. This phenotype was rescued upon co-injection of human mRNA together along with the trim69 knockdown. Results of this study also showed an interaction between TRIM69 and c-Jun in human cells, and upon TRIM69 knock down c-Jun expression subsequently increased, whereas the over-expression of TRIM69 led to the down-regulation of c-Jun. Additionally, knockdown both c-Jun and trim69 can rescue the deformed brain, evident cellular apoptosis in the head and decreased expression of neuronal differentiation and stem cell markers. Overall, our results support a role for trim69 in the development of the zebrafish brain through ap-1 pathway. PMID:27050765

  15. Pulsed power supply for three APS septum magnets

    SciTech Connect

    McGhee, D.G.

    1991-03-24

    Three septum magnets will be operated at a repetition-rate of 2 Hz. Two of the septum magnets are identical and operate at the same values; these are the synchrotron extraction and the storage ring injection magnets. They are transformer septum magnets, with a primary inductance of 23 {mu}H and resistance of 6.3 m{Omega}, and must be pulsed at a 2 Hz rate to extract beam from the synchrotron and inject beam into the storage ring at 7.7 GeV. The third septum magnet is used to inject electrons into the synchrotron at 650 MeV or positrons at 450 MeV. It is also a transformer septum magnet, with a primary inductance of 21 {mu}H and resistance of 6.7 m{Omega}, and must be pulsed at a 2 Hz rate. A design study was performed of the power supply proposed in the APS Title I design. This supply produces a pulse that is approximately a half-sine-wave with a base width of approximately 1/3 ms; its peakcurrent is adjustable from 470 A to 4.7 kA and is repeatable within {plus_minus}0.05%. The septum steel is reset by a half-sine pulse of reverse polarity a few milliseconds after the forward current pulse. No beam is present during reset. The use of the transformer design minimizes the cost of the capacitors used for energy storage.

  16. Measurements of the electron cloud in the APS storage ring.

    SciTech Connect

    Harkey, K. C.

    1999-04-16

    Synchrotron radiation interacting with the vacuum chamber walls in a storage ring produce photoelectrons that can be accelerated by the beam, acquiring sufficient energy to produce secondary electrons in collisions with the walls. If the secondary-electron yield (SEY) coefficient of the wall material is greater than one, as is the case with the aluminum chambers in the Advanced Photon Source (APS) storage ring, a runaway condition can develop. As the electron cloud builds up along a train of stored positron or electron bunches, the possibility exists that a transverse perturbation of the head bunch will be communicated to trailing bunches due to interaction with the cloud. In order to characterize the electron cloud, a special vacuum chamber was built and inserted into the ring. The chamber contains 10 rudimentary electron-energy analyzers, as well as three targets coated with different materials. Measurements show that the intensity and electron energy distribution are highly dependent on the temporal spacing between adjacent bunches and the amount of current contained in each bunch. Furthermore, measurements using the different targets are consistent with what would be expected based on the SEY of the coatings. Data for both positron and electron beams are presented.

  17. A quantum network with atoms and photons (QNET-AP)

    NASA Astrophysics Data System (ADS)

    Meyers, Ronald E.; Lee, Patricia; Deacon, Keith S.; Tunick, Arnold; Quraishi, Qudsia; Stack, Daniel

    2012-10-01

    Enabling secure communication, unparalleled computing capabilities, and fundamental nonlocality physics exploration, the development of quantum repeaters is the key quantum information processing technology advance needed for implementing real world quantum networks beyond the laboratory environment. Currently, components exist for intra-laboratory quantum networks but no system exists for connecting distant ( 1 km ) quantum memories in the real world. We present a physics analysis of quantum repeater network designs for intracity optical fiber connections between nodes based on atomic memories and linear optics. Long distances will necessitate the use of (1) two-photon Hong-Ou-Mandel style interference between atomic ensembles for entanglement swapping, and (2) photonic qubit wavelength conversion between atomic emissions and photons at telecommunication wavelengths in fiber. We report on our experimental progress towards implementing A Quantum Network with Atoms and Photons (QNET-AP), a quantum repeater network test-bed, between the US Army Research Laboratory (ARL) and the Joint Quantum Institute (JQI) of the National Institute of Standards and Technology (NIST) and the University of Maryland (UMD).

  18. Absolute properties of the eclipsing binary star AP Andromedae

    SciTech Connect

    Sandberg Lacy, Claud H.; Torres, Guillermo; Fekel, Francis C.; Muterspaugh, Matthew W. E-mail: gtorres@cfa.harvard.edu E-mail: matthew1@coe.tsuniv.edu

    2014-06-01

    AP And is a well-detached F5 eclipsing binary star for which only a very limited amount of information was available before this publication. We have obtained very extensive measurements of the light curve (19,097 differential V magnitude observations) and a radial velocity curve (83 spectroscopic observations) which allow us to fit orbits and determine the absolute properties of the components very accurately: masses of 1.277 ± 0.004 and 1.251 ± 0.004 M {sub ☉}, radii of 1.233 ± 0.006 and 1.1953 ± 0.005 R {sub ☉}, and temperatures of 6565 ± 150 K and 6495 ± 150 K. The distance to the system is about 400 ± 30 pc. Comparison with the theoretical properties of the stellar evolutionary models of the Yonsei-Yale series of Yi et al. shows good agreement between the observations and the theory at an age of about 500 Myr and a slightly sub-solar metallicity.

  19. Global coupling and decoupling of the APS storage ring

    SciTech Connect

    Chae, Yong-Chul; Liu, Jianyang; Teng, L.C.

    1995-07-01

    This Paper describes a study of controlling the coupling between the horizontal and the vertical betatron oscillations in the APS storage ring. First, we investigate the strengthening of coupling using two families of skew quadrupoles. Using smooth approximation, we obtained the formulae to estimate the coupling ratio defined as the ratio of the vertical and horizontal emittances or, for a single particle, the ratio of the maximum values of the Courant Snyder invariants. Since we knew that the coupling is mostly enhanced by the 21st harmonic content of skew quadrupole distribution, we carried out the harmonic analysis in order to find the optimum arrangement of the skew quadrupoles. The numerical results from tracking a single particle are presented for the various configurations of skew quadrupoles. Second, we describe the global decoupling procedure to minimize the unwanted coupling effects. These are mainly due to the random roll errors of normal quadrupoles. It is shown that even with the rather large rms roll error of 2 mrad we can reduce the Coupling from 70 percent to 10 percent with a skew quadrupole strength which is one order of magnitude lower than the typical normal quadrupole strength.

  20. Scribble1/AP2 complex coordinates NMDA receptor endocytic recycling.

    PubMed

    Piguel, Nicolas H; Fievre, Sabine; Blanc, Jean-Michel; Carta, Mario; Moreau, Maïté M; Moutin, Enora; Pinheiro, Vera L; Medina, Chantal; Ezan, Jerome; Lasvaux, Léa; Loll, François; Durand, Christelle M; Chang, Kai; Petralia, Ronald S; Wenthold, Robert J; Stephenson, F Anne; Vuillard, Laurent; Darbon, Hervé; Perroy, Julie; Mulle, Christophe; Montcouquiol, Mireille; Racca, Claudia; Sans, Nathalie

    2014-10-23

    The appropriate trafficking of glutamate receptors to synapses is crucial for basic synaptic function and synaptic plasticity. It is now accepted that NMDA receptors (NMDARs) internalize and are recycled at the plasma membrane but also exchange between synaptic and extrasynaptic pools; these NMDAR properties are also key to governing synaptic plasticity. Scribble1 is a large PDZ protein required for synaptogenesis and synaptic plasticity. Herein, we show that the level of Scribble1 is regulated in an activity-dependent manner and that Scribble1 controls the number of NMDARs at the plasma membrane. Notably, Scribble1 prevents GluN2A subunits from undergoing lysosomal trafficking and degradation by increasing their recycling to the plasma membrane following NMDAR activation. Finally, we show that a specific YxxR motif on Scribble1 controls these mechanisms through a direct interaction with AP2. Altogether, our findings define a molecular mechanism to control the levels of synaptic NMDARs via Scribble1 complex signaling. PMID:25310985