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Sample records for apurinic sites design

  1. Response of phage T4 polynucleotide kinase toward dinucleotides containing apurinic sites: Design of a sup 32 P-postlabeling assay for apurinic sites in DNA

    SciTech Connect

    Weinfeld, M.; Liuzzi, M.; Paterson, M.C. )

    1990-02-20

    The authors have examined the capacity of bacteriophage T4 polynucleotide kinase to phosphorylate the partially depurinated products of d-ApA, namely d-SpA and d-ApS (where S represents an apurinic deoxyribose group). It was observed that the enzyme acted only on the latter isomer. Since molecules of this type (d-NpS) are the sole apurinic site containing products resulting from the combined digestion of lightly depurinated DNA by snake venom phosphodiesterase and calf alkaline phosphatase they were able to devise a postlabeling assay for these biologically important DNA lesions. The method offers several advantages, including (a) elimination of the need for prelabeled DNA, (b) high (femtomole range) sensitivity, and (c) nearest-neighbor analysis of bases 5{prime} to apurinic/apyrimidinic sites. Using this assay, they obtained a value for the rate of depurination of form I pRSV neo plasmid DNA. The rate of depurination of poly(dA), treated in a similar fashion, was found to be {approximately}1 base per 10{sup 3} nucleotides per hour.

  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. Excision of apurinic sites from DNA with enzymes isolated from rat-liver chromatin.

    PubMed

    Goffin, C; Verly, W G

    1982-10-01

    Apurinic sites were excised from phi X174 RF DNA with two enzymes isolated from rat liver chromatin: an apurinic/apyrimidinic endodeoxyribonuclease and a 5'-3'-exonuclease; the resulting gap was filled with DNA polymerase beta also prepared from rat liver chromatin and the repair was fully terminated with T4 ligase.

  4. Modified alkaline elution allows the measurement of intact apurinic sites in mammalian genomic DNA.

    PubMed

    Malvy, C; Lefrançois, M; Bertrand, J R; Markovits, J

    2000-08-01

    The presence of apurinic/apyrimidinic (AP) sites in cell genomes is known to be toxic and mutagenic. These lesions are therefore repaired in cells by efficient enzymatic systems. However, a report (Nakamura and Swenberg, Cancer Res. 59 (1999) 2522-2526) indicates an unexpected high rate of endogenous apurinic/apyrimidinic (AP) sites in genomic DNA in mammalian tissues. The technology used does not allow the authors to distinguish between intact AP sites and 3'cleaved AP sites. The corresponding values range between 2 and 4 sites per million of nucleotides in various human and rat tissues. Using a modified alkaline elution method we show here that the stationary level of intact AP sites is about 0.16 per million of nucleotides in leukemic mouse L1210 cells.

  5. Design of a New Fluorescent Oligonucleotide-Based Assay for a Highly Specific Real-Time Detection of Apurinic/Apyrimidinic Site Cleavage by Tyrosyl-DNA Phosphodiesterase 1.

    PubMed

    Lebedeva, Natalia A; Anarbaev, Rashid O; Kupryushkin, Maxim S; Rechkunova, Nadejda I; Pyshnyi, Dmitrii V; Stetsenko, Dmitry A; Lavrik, Olga I

    2015-10-21

    Tyrosyl-DNA phosphodiesterase 1 (Tdp1) promotes catalytic scission of a phosphodiester bond between the 3'-end of DNA and the hydroxyl group of a tyrosine residue, as well as cleaving off a variety of other 3'-terminal phosphate-linked DNA substituents. We have shown recently that Tdp1 can initiate an apurinic/apyrimidinic (AP) site repair pathway that is independent from the one mediated by AP endonuclease 1 (APE1). Until recently, there was no method available of tracking the AP-site cleaving activity of Tdp1 by real-time fluorescence assay. In the present study we demonstrate a highly specific real-time detection of the AP-site cleaving activity of Tdp1 which allows one to distinguish it from the activity of APE1 by using a short hairpin oligonucleotide with a 1,12-dodecanediol loop, a 5'-fluorophore, and a 3'-quencher. Specific phosphodiesterase activity of Tdp1, which is usually able to remove quencher from the 3'-end of DNA, was suppressed in our approach by introducing a noncleavable phosphate group mimic between the 3'-end and the quencher. As a nondigestible 3'-phosphate analogue, we have used a new uncharged tetramethyl phosphoryl guanidine (Tmg) group, which is resistant to 3'-phosphodiesterase cleavage.

  6. Characterization of the endoribonuclease active site of human apurinic/apyrimidinic endonuclease 1.

    PubMed

    Kim, Wan-Cheol; Berquist, Brian R; Chohan, Manbir; Uy, Christopher; Wilson, David M; Lee, Chow H

    2011-09-02

    Apurinic/apyrimidinic endonuclease 1 (APE1) is the major mammalian enzyme in DNA base excision repair that cleaves the DNA phosphodiester backbone immediately 5' to abasic sites. Recently, we identified APE1 as an endoribonuclease that cleaves a specific coding region of c-myc mRNA in vitro, regulating c-myc mRNA level and half-life in cells. Here, we further characterized the endoribonuclease activity of APE1, focusing on the active-site center of the enzyme previously defined for DNA nuclease activities. We found that most site-directed APE1 mutant proteins (N68A, D70A, Y171F, D210N, F266A, D308A, and H309S), which target amino acid residues constituting the abasic DNA endonuclease active-site pocket, showed significant decreases in endoribonuclease activity. Intriguingly, the D283N APE1 mutant protein retained endoribonuclease and abasic single-stranded RNA cleavage activities, with concurrent loss of apurinic/apyrimidinic (AP) site cleavage activities on double-stranded DNA and single-stranded DNA (ssDNA). The mutant proteins bound c-myc RNA equally well as wild-type (WT) APE1, with the exception of H309N, suggesting that most of these residues contributed primarily to RNA catalysis and not to RNA binding. Interestingly, both the endoribonuclease and the ssRNA AP site cleavage activities of WT APE1 were present in the absence of Mg(2+), while ssDNA AP site cleavage required Mg(2+) (optimally at 0.5-2.0 mM). We also found that a 2'-OH on the sugar moiety was absolutely required for RNA cleavage by WT APE1, consistent with APE1 leaving a 3'-PO(4)(2-) group following cleavage of RNA. Altogether, our data support the notion that a common active site is shared for the endoribonuclease and other nuclease activities of APE1; however, we provide evidence that the mechanisms for cleaving RNA, abasic single-stranded RNA, and abasic DNA by APE1 are not identical, an observation that has implications for unraveling the endoribonuclease function of APE1 in vivo. Copyright

  7. Relating aromatic hydrocarbon-induced DNA adducts and c-H-ras mutations in mouse skin papillomas: the role of apurinic sites.

    PubMed Central

    Chakravarti, D; Pelling, J C; Cavalieri, E L; Rogan, E G

    1995-01-01

    Mouse skin tumors contain activated c-H-ras oncogenes, often caused by point mutations at codons 12 and 13 in exon 1 and codons 59 and 61 in exon 2. Mutagenesis by the noncoding apurinic sites can produce G-->T and A-->T transversions by DNA misreplication with more frequent insertion of deoxyadenosine opposite the apurinic site. Papillomas were induced in mouse skin by several aromatic hydrocarbons, and mutations in the c-H-ras gene were determined to elucidate the relationship among DNA adducts, apurinic sites, and ras oncogene mutations. Dibenzo[a,l]pyrene (DB[a,l]P), DB[a,l]P-11,12-dihydrodiol, anti-DB[a,l]P-11,12-diol-13,14-epoxide, DB[a,l]P-8,9-dihydrodiol, 7,12-dimethylbenz[a]anthracene (DMBA), and 1,2,3,4-tetrahydro-DMBA consistently induced a CAA-->CTA mutation in codon 61 of the c-H-ras oncogene. Benzo[a]pyrene induced a GGC-->GTC mutation in codon 13 in 54% of tumors and a CAA-->CTA mutation in codon 61 in 15%. The pattern of mutations induced by each hydrocarbon correlated with its profile of DNA adducts. For example, both DB[a,l]P and DMBA primarily form DNA adducts at the N-3 and/or N-7 of deoxyadenosine that are lost from the DNA by depurination, generating apurinic sites. Thus, these results support the hypothesis that misreplication of unrepaired apurinic sites generated by loss of hydrocarbon-DNA adducts is responsible for transforming mutations leading to papillomas in mouse skin. PMID:7479797

  8. Transient-state kinetics of apurinic/apyrimidinic (AP) endonuclease 1 acting on an authentic AP site and commonly used substrate analogs: the effect of diverse metal ions and base mismatches.

    PubMed

    Schermerhorn, Kelly M; Delaney, Sarah

    2013-10-29

    Apurinic/apyrimidinic endonuclease 1 (APE1) is an Mg²⁺-dependent enzyme responsible for incising the DNA backbone 5' to an apurinic/apyrimidinic (AP) site. Here, we use rapid quench flow (RQF) techniques to provide a comprehensive kinetic analysis of the strand-incision activity (k(chemistry)) of APE1 acting on an authentic AP site along with two widely used analogs, a reduced AP site and a tetrahydrofuran (THF) site. In the presence of biologically relevant Mg²⁺, APE1 incises all three substrates at a rate faster than the resolution of the RQF, ≥700 s⁻¹. To obtain quantitative values of k(chemistry) and to facilitate a comparison of the authentic substrate versus the substrate analogs, we replaced Mg²⁺ with Mn²⁺ or Ni²⁺ or introduced a mismatch 5' to the lesion site. Both strategies were sufficient to slow k(chemistry) and resulted in rates within the resolution of the RQF. In all cases where quantitative rates were obtained, k(chemistry) for the reduced AP site is indistinguishable from the authentic AP site. Notably, there is a small decrease, ~1.5-fold, in k(chemistry) for the THF site relative to the authentic AP site. These results highlight a role in strand incision for the C1' oxygen of the AP site and warrant consideration when designing experiments using substrate analogs.

  9. Identification of one of the apurinic/apyrimidinic lyase active sites of topoisomerase V by structural and functional studies

    PubMed Central

    Rajan, Rakhi; Prasad, Rajendra; Taneja, Bhupesh; Wilson, Samuel H.; Mondragón, Alfonso

    2013-01-01

    Topoisomerase V (Topo-V) is the only member of a novel topoisomerase subtype. Topo-V is unique because it is a bifunctional enzyme carrying both topoisomerase and DNA repair lyase activities within the same protein. Previous studies had shown that the topoisomerase domain spans the N-terminus of the protein and is followed by 12 tandem helix–hairpin–helix [(HhH)2] domains. There are at least two DNA repair lyase active sites for apurinic/apyrimidinic (AP) site processing, one within the N-terminal region and the second within the C-terminal domain of Topo-V, but their exact locations and characteristics are unknown. In the present study, the N-terminal 78-kDa fragment of Topo-V (Topo-78), containing the topoisomerase domain and one of the lyase DNA repair domains, was characterized by structural and biochemical studies. The results show that an N-terminal 69-kDa fragment is the minimal fragment with both topoisomerase and AP lyase activities. The lyase active site of Topo-78 is at the junction of the fifth and sixth (HhH)2 domains. From the biochemical and structural data, it appears that Lys571 is the most probable nucleophile responsible for the lyase activity. Our experiments also suggest that Topo-V most likely acts as a Class I AP endonuclease in vivo. PMID:23125368

  10. High-resolution crystal structures reveal plasticity in the metal binding site of apurinic/apyrimidinic endonuclease I.

    PubMed

    He, Hongzhen; Chen, Qiujia; Georgiadis, Millie M

    2014-10-21

    Apurinic/apyrimidinic endonuclease I (APE1) is an essential base excision repair enzyme that catalyzes a Mg²⁺-dependent reaction in which the phosphodiester backbone is cleaved 5' of an abasic site in duplex DNA. This reaction has been proposed to involve either one or two metal ions bound to the active site. In the present study, we report crystal structures of Mg²⁺, Mn²⁺, and apo-APE1 determined at 1.4, 2.2, and 1.65 Å, respectively, representing two of the highest resolution structures yet reported for APE1. In our structures, a single well-ordered Mn²⁺ ion was observed coordinated by D70 and E96; the Mg²⁺ site exhibited disorder modeled as two closely positioned sites coordinated by D70 and E96 or E96 alone. Direct metal binding analysis of wild-type, D70A, and E96A APE1, as assessed by differential scanning fluorimetry, indicated a role for D70 and E96 in binding of Mg²⁺ or Mn²⁺ to APE1. Consistent with the disorder exhibited by Mg²⁺ bound to the active site, two different conformations of E96 were observed coordinated to Mg²⁺. A third conformation for E96 in the apo structure is similar to that observed in the APE1-DNA-Mg²⁺ complex structure. Thus, binding of Mg²⁺ in three different positions within the active site of APE1 in these crystal structures corresponds directly with three different conformations of E96. Taken together, our results are consistent with the initial capture of metal by D70 and E96 and repositioning of Mg²⁺ facilitated by the structural plasticity of E96 in the active site.

  11. Interaction of Nucleotide Excision Repair Protein XPC-RAD23B with DNA Containing Benzo[a]pyrene-Derived Adduct and Apurinic/Apyrimidinic Site within a Cluster.

    PubMed

    Starostenko, L V; Maltseva, E A; Lebedeva, N A; Pestryakov, P E; Lavrik, O I; Rechkunova, N I

    2016-03-01

    The combined action of reactive metabolites of benzo[a]pyrene (B[a]P) and oxidative stress can lead to cluster-type DNA damage that includes both a bulky lesion and an apurinic/apyrimidinic (AP) site, which are repaired by the nucleotide and base excision repair mechanisms - NER and BER, respectively. Interaction of NER protein XPC-RAD23B providing primary damage recognition with DNA duplexes containing a B[a]P-derived residue linked to the exocyclic amino group of a guanine (BPDE-N(2)-dG) in the central position of one strand and AP site in different positions of the other strand was analyzed. It was found that XPC-RAD23B crosslinks to DNA containing (+)-trans-BPDE-N(2)-dG more effectively than to DNA containing cis-isomer, independently of the AP site position in the opposite strand; protein affinity to DNA containing one of the BPDE-N(2)-dG isomers depends on the AP site position in the opposite strand. The influence of XPC-RAD23B on hydrolysis of an AP site clustered with BPDE-N(2)-dG catalyzed by the apurinic/apyrimidinic endonuclease 1 (APE1) was examined. XPC-RAD23B was shown to stimulate the endonuclease and inhibit the 3'-5' exonuclease activity of APE1. These data demonstrate the possibility of cooperation of two proteins belonging to different DNA repair systems in the repair of cluster-type DNA damage.

  12. Loss of an apurinic/apyrimidinic site endonuclease increases the mutagencity of N-methyl-N'-nitro-N-nitrosoguanidine to Escherichia coli

    SciTech Connect

    Foster, P.L.; Davis, E.F.

    1987-05-01

    xthA/sup -/ Escherichia coli, which are missing a major cellular apurinic/apyrimidinic (AP) endonuclease, are 5- to 10-fold more sensitive than xthA/sup +/ bacteria to mutagenesis by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) under conditions that induce the adaptive response. The xthA/sup -/-dependent mutations are also dependent on SOS mutagenic processing and consists of both transversion and transition base substitutions. When MNNG-adapted xthA/sup -/ bacteria are challenged with a high dose of MNNG, more xthA/sup -/-dependent SOS-dependent mutations are induced, and transversions are enhanced relative to transitions. The mutations induced by challenge are eliminated in xthA/sup -/ alkA/sup -/ bacteria, which are also deficient for 3-methyladenine glycosylase II activity. these data are consistent with the hypothesis that AP sites, at least some of which are produced by glycosylase activity, are mutagenic intermediates following cellular DNA alkylation.

  13. Human Apurinic/Apyrimidinic Endonuclease (APE1) Is Acetylated at DNA Damage Sites in Chromatin, and Acetylation Modulates Its DNA Repair Activity

    PubMed Central

    Roychoudhury, Shrabasti; Nath, Somsubhra; Song, Heyu; Hegde, Muralidhar L.; Bellot, Larry J.; Mantha, Anil K.; Sengupta, Shiladitya; Ray, Sutapa; Natarajan, Amarnath

    2016-01-01

    ABSTRACT Apurinic/apyrimidinic (AP) sites, the most frequently formed DNA lesions in the genome, inhibit transcription and block replication. The primary enzyme that repairs AP sites in mammalian cells is the AP endonuclease (APE1), which functions through the base excision repair (BER) pathway. Although the mechanism by which APE1 repairs AP sites in vitro has been extensively investigated, it is largely unknown how APE1 repairs AP sites in cells. Here, we show that APE1 is acetylated (AcAPE1) after binding to the AP sites in chromatin and that AcAPE1 is exclusively present on chromatin throughout the cell cycle. Positive charges of acetylable lysine residues in the N-terminal domain of APE1 are essential for chromatin association. Acetylation-mediated neutralization of the positive charges of the lysine residues in the N-terminal domain of APE1 induces a conformational change; this in turn enhances the AP endonuclease activity of APE1. In the absence of APE1 acetylation, cells accumulated AP sites in the genome and showed higher sensitivity to DNA-damaging agents. Thus, mammalian cells, unlike Saccharomyces cerevisiae or Escherichia coli cells, require acetylation of APE1 for the efficient repair of AP sites and base damage in the genome. Our study reveals that APE1 acetylation is an integral part of the BER pathway for maintaining genomic integrity. PMID:27994014

  14. Apurinic/apyrimidinic (AP) site recognition by the 5'-dRP/AP lyase in poly(ADP-ribose) polymerase-1 (PARP-1).

    PubMed

    Khodyreva, S N; Prasad, R; Ilina, E S; Sukhanova, M V; Kutuzov, M M; Liu, Y; Hou, E W; Wilson, S H; Lavrik, O I

    2010-12-21

    The capacity of human poly(ADP-ribose) polymerase-1 (PARP-1) to interact with intact apurinic/apyrimidinic (AP) sites in DNA has been demonstrated. In cell extracts, sodium borohydride reduction of the PARP-1/AP site DNA complex resulted in covalent cross-linking of PARP-1 to DNA; the identity of cross-linked PARP-1 was confirmed by mass spectrometry. Using purified human PARP-1, the specificity of PARP-1 binding to AP site-containing DNA was confirmed in competition binding experiments. PARP-1 was only weakly activated to conduct poly(ADP-ribose) synthesis upon binding to AP site-containing DNA, but was strongly activated for poly(ADP-ribose) synthesis upon strand incision by AP endonuclease 1 (APE1). By virtue of its binding to AP sites, PARP-1 could be poised for its role in base excision repair, pending DNA strand incision by APE1 or the 5'-dRP/AP lyase activity in PARP-1.

  15. 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

  16. Efficacy of puffer fish (Takifugu rubripes) sauce in reducing hydroxyl radical damage to DNA assessed using the apurinic/apyrimidinic site method.

    PubMed

    Harada, Kazuki; Makino, Yoshio; Yamauchi, Tomio; Fukuda, Nami; Tamaru, Miki; Okubo, Yasue; Maeda, Toshimichi; Fukuda, Yutaka; Shiba, Tsuneo

    2007-09-01

    Apurinic/apyrimidinic (AP) sites are frequently observed DNA lesions when cells are exposed to hydroxyl radicals. We developed a new method for measurement of the antioxidative activity of foods using the occurrence frequency of AP sites on DNA. Combined with the electron spin resonance (ESR) method as a standard method, we examined whether fish and soy sauces including puffer fish [Takifugu rubripes (Temminck et Schlegel)] sauce could protect DNA from damage caused by hydroxyl radicals. The results showed that the ratios of DNA protection by puffer fish sauce, salmon fish sauce, sandfish fish sauce (Shottsuru), colorless soy sauce, squid fish sauce (Ishiru), dark color soy sauce and light color soy sauce were 68.9, 67.0, 60.1, 49.7, 34.1, 28.2 and -4.4%, respectively. Puffer, salmon, and sandfish fish sauces showed high ratios of DNA protection against hydroxyl radicals. On the other hand, IC(50) values of hydroxyl radical scavenging of the puffer, salmon, sandfish, squid fish sauces and colorless, dark and light color soy sauces were 0.20, 0.09, 4.16, 0.26% and 0.28, 0.14 and 0.18%, respectively. Though the puffer fish sauce exhibited the highest level of DNA protection among the examined samples and a high hydroxyl radical scavenging capability, a correlation between the radical scavenging capability and DNA protection against hydroxyl radicals among the examined fish and soy sauces was not found.

  17. DDB1-DDB2 (xeroderma pigmentosum group E) protein complex recognizes a cyclobutane pyrimidine dimer, mismatches, apurinic/apyrimidinic sites, and compound lesions in DNA.

    PubMed

    Wittschieben, Birgitte Ø; Iwai, Shigenori; Wood, Richard D

    2005-12-02

    The DDB protein complex, comprising the subunits DDB1 and DDB2, binds tightly to UV light-irradiated DNA. Mutations in DDB2 are responsible for xeroderma pigmentosum group E, a disorder with defects in nucleotide excision repair of DNA. Both subunits are also components of a complex involved in ubiquitin-mediated proteolysis. Cellular defects in DDB2 disable repair of the major UV radiation photoproduct in DNA, a cyclobutane pyrimidine dimer, but no significant direct binding of DDB to this photoproduct in DNA has ever been demonstrated. Thus, it has been uncertain how DDB could play a specific role in DNA repair of such damage. We investigated DDB function using highly purified proteins. Co-purified DDB1-DDB2 or DDB reconstituted with individual DDB1 and DDB2 subunits binds to damaged DNA as a ternary complex. We found that DDB can indeed recognize a cyclobutane pyrimidine dimer in DNA with an affinity (K(app)a) 6-fold higher than that of nondamaged DNA. The DDB1-DDB2 complex also bound with high specificity to a UV radiation-induced (6-4) photoproduct and to an apurinic site in DNA. Unexpectedly, DDB also bound avidly to DNA containing a 2- or 3-bp mismatch (and does not bind well to DNA containing larger mismatches). These data indicate that DDB does not detect lesions per se. It instead recognizes other structural features of damaged DNA, acting as a sensor that probes DNA for a subset of conformational changes. Lesions recognized may include those arising when translesion polymerases such as POLH incorporate bases across from DNA lesions caused by UV radiation.

  18. Quantitation of DNA and hemoglobin adducts and apurinic/apyrimidinic sites in tissues of F344 rats exposed to propylene oxide by inhalation.

    PubMed

    Ríos-Blanco, M N; Faller, T H; Nakamura, J; Kessler, W; Kreuzer, P E; Ranasinghe, A; Filser, J G; Swenberg, J A

    2000-11-01

    Propylene oxide (PO) is a relatively weak mutagen that induces nasal tumor formation in rats during long-term inhalation studies at high exposures (> or =300 p.p.m.), concentrations that also cause cytotoxicity and increases in cell proliferation. Direct alkylation of DNA by PO leads mainly to the formation of N:7-(2-hydroxypropyl)guanine (7-HPG). In this study, the accumulation of 7-HPG in tissues of male F344 rats exposed to 500 p. p.m. PO (6 h/day, 5 days/week for 4 weeks) by the inhalation route was measured by gas chromatography-high resolution mass spectrometry (GC-HRMS). In animals killed up to 7 h following the end of the last exposure the levels of 7-HPG (pmol/micromol guanine) in nasal respiratory tissue, nasal olfactory tissue, lung, spleen, liver and testis DNA were 606.2 +/- 53.0, 297.5 +/- 56.5, 69.8 +/- 3.8, 43.0 +/- 3.8, 27.5 +/- 2.4 and 14.2 +/- 0.7, respectively. The amounts of 7-HPG in the same tissues of animals killed 3 days after cessation of exposure were 393.3 +/- 57.0, 222.7 +/- 29.5, 51.5 +/- 1.2, 26.7 +/- 1.0, 18.0 +/- 2.6 and 10.4 +/- 0.1. A comparable rate of disappearance of 7-HPG was found among all tissues. DNA from lymphocytes pooled from four rats killed at the end of the last exposure was found to have 39.6 pmol adduct/micromol guanine. Quantitation of DNA apurinic/apyrimidinic sites, potentially formed after adduct loss by chemical depurination or DNA repair, showed no difference between tissues from control and exposed rats. The level of N:-(2-hydroxypropyl)valine in hemoglobin of exposed rats was also determined using a modified Edman degradation method followed by GC-HRMS analysis. The value obtained was 90.2 +/- 10.3 pmol/mg globin. These data demonstrate that nasal respiratory tissue, which is the target tissue for carcinogenesis, has a much greater level of alkylation of DNA than non-target tissues.

  19. Effects of ethylene oxide and ethylene inhalation on DNA adducts, apurinic/apyrimidinic sites and expression of base excision DNA repair genes in rat brain, spleen, and liver.

    PubMed

    Rusyn, Ivan; Asakura, Shoji; Li, Yutai; Kosyk, Oksana; Koc, Hasan; Nakamura, Jun; Upton, Patricia B; Swenberg, James A

    2005-09-28

    Ethylene oxide (EO) is an important industrial chemical that is classified as a known human carcinogen (IARC, Group 1). It is also a metabolite of ethylene (ET), a compound that is ubiquitous in the environment and is the most used petrochemical. ET has not produced evidence of cancer in laboratory animals and is "not classifiable as to its carcinogenicity to humans" (IARC, Group 3). The mechanism of carcinogenicity of EO is not well characterized, but is thought to involve the formation of DNA adducts. EO is mutagenic in a variety of in vitro and in vivo systems, whereas ET is not. Apurinic/apyrimidinic sites (AP) that result from chemical or glycosylase-mediated depurination of EO-induced DNA adducts could be an additional mechanism leading to mutations and chromosomal aberrations. This study tested the hypothesis that EO exposure results in the accumulation of AP sites and induces changes in expression of genes for base excision DNA repair (BER). Male Fisher 344 rats were exposed to EO (100 ppm) or ET (40 or 3000 ppm) by inhalation for 1, 3 or 20 days (6h/day, 5 days a week). Animals were sacrificed 2h after exposure for 1, 3 or 20 days as well as 6, 24 and 72 h after a single-day exposure. Experiments were performed with tissues from brain and spleen, target sites for EO-induced carcinogenesis, and liver, a non-target organ. Exposure to EO resulted in time-dependent increases in N7-(2-hydroxyethyl)guanine (7-HEG) in brain, spleen, and liver and N7-(2-hydroxyethyl)valine (7-HEVal) in globin. Ethylene exposure also induced 7-HEG and 7-HEVal, but the numbers of adducts were much lower. No increase in the number of aldehydic DNA lesions, an indicator of AP sites, was detected in any of the tissues between controls and EO-, or ET-exposed animals, regardless of the duration or strength of exposure. EO exposure led to a 3-7-fold decrease in expression of 3-methyladenine-DNA glycosylase (Mpg) in brain and spleen in rats exposed to EO for 1 day. Expression of 8

  20. Physical association of the 2,6-diamino-4-hydroxy-5N-formamidopyrimidine-DNA glycosylase of Escherichia coli and an activity nicking DNA at apurinic/apyrimidinic sites.

    PubMed Central

    O'Connor, T R; Laval, J

    1989-01-01

    The 2,6-diamino-4-hydroxy-5N-formamidopyrimidine (Fapy)-DNA glycosylase of Escherichia coli, which is coded for by the fpg gene, excises purine bases with ring-opened imidazoles. In addition to the DNA glycosylase activity, we report that the Fapy-DNA glycosylase of E. coli has an associated activity, resistant to EDTA, that nicks DNA at apurinic/apyrimidinic (AP) sites. The levels of Fapy-DNA glycosylase and AP-nicking activity were parallel in crude lysates of E. coli HB101 harboring different plasmids constructed from the fpg gene. The fpg gene is different from the xth, nth, and nfo genes of E. coli, whose gene products also cleave DNA at AP sites. The Fapy-DNA glycosylase was purified to electrophoretic homogeneity. During this purification, the Fapy-DNA glycosylase copurified with an AP-nicking activity using chromatographic separations based on ion-exchange, molecular weight exclusion, and hydrophobicity. The cleavage at AP sites by the Fapy-DNA glycosylase left a 5'-phosphomonoester nucleotide at one terminus. In addition, DNA containing reduced AP sites was not nicked by the Fapy-DNA glycosylase. These data suggest that the mechanism of cleavage involved beta elimination. Therefore, this activity of the Fapy-DNA glycosylase nicking DNA at AP sites should be referred to as an AP lyase. The 3' terminus did not prime nick-translation by E. coli DNA polymerase I. However, the 3' terminus becomes a substrate for nick-translation if first allowed to react with calf intestine phosphatase or the E. coli exonuclease III. These data suggest that the repair of the Fapy lesion at least to some extent results in the formation of both 5'- and 3'-phosphomonoester nucleotides and the release of the deoxyribose. Images PMID:2664776

  1. Human Apurinic/Apyrimidinic Endonuclease 1

    PubMed Central

    Li, Mengxia

    2014-01-01

    Abstract Significance: Human apurinic/apyrimidinic endonuclease 1 (APE1, also known as REF-1) was isolated based on its ability to cleave at AP sites in DNA or activate the DNA binding activity of certain transcription factors. We review herein topics related to this multi-functional DNA repair and stress-response protein. Recent Advances: APE1 displays homology to Escherichia coli exonuclease III and is a member of the divalent metal-dependent α/β fold-containing phosphoesterase superfamily of enzymes. APE1 has acquired distinct active site and loop elements that dictate substrate selectivity, and a unique N-terminus which at minimum imparts nuclear targeting and interaction specificity. Additional activities ascribed to APE1 include 3′–5′ exonuclease, 3′-repair diesterase, nucleotide incision repair, damaged or site-specific RNA cleavage, and multiple transcription regulatory roles. Critical Issues: APE1 is essential for mouse embryogenesis and contributes to cell viability in a genetic background-dependent manner. Haploinsufficient APE1+/− mice exhibit reduced survival, increased cancer formation, and cellular/tissue hyper-sensitivity to oxidative stress, supporting the notion that impaired APE1 function associates with disease susceptibility. Although abnormal APE1 expression/localization has been seen in cancer and neuropathologies, and impaired-function variants have been described, a causal link between an APE1 defect and human disease remains elusive. Future Directions: Ongoing efforts aim at delineating the biological role(s) of the different APE1 activities, as well as the regulatory mechanisms for its intra-cellular distribution and participation in diverse molecular pathways. The determination of whether APE1 defects contribute to human disease, particularly pathologies that involve oxidative stress, and whether APE1 small-molecule regulators have clinical utility, is central to future investigations. Antioxid. Redox Signal. 20, 678–707

  2. Apurinic/apyrimidinic endonucleases in repair of pyrimidine dimers and other lesions in DNA.

    PubMed Central

    Warner, H R; Demple, B F; Deutsch, W A; Kane, C M; Linn, S

    1980-01-01

    The characteristics of the nicks (single-strand breaks) introduced into damaged DNA by Escherichia coli endonucleases III, IV, and VI and by phage T4 UV endonuclease have been investigated with E. coli DNA polymerase I (DNA nucleotidyltransferase). Nicks introduced into depurinated DNA by endonuclease IV or VI provide good primer termini for the polymerase, whereas nicks introduced into depurinated DNA by endonuclease III or into irradiated DNA by T4 UV endonuclease do not. This result suggests that endonuclease IV nicks depurinated DNA on the 5' side of the apurinic site, as does endonuclease VI, whereas endonuclease III has a different incision mechanism. T4 UV endonuclease also possesses apurinic endonuclease activity that generates nicks in depurinated DNA with low priming activity for the polymerase. The priming activity of DNA nicked with endonuclease III or T4 UV endonuclease can be enhanced by an additional incubation with endonuclease VI and, to a lesser extent, by incubation with endonuclease IV. These results indicate that endonuclease III and T4 UV endonuclease (acting upon depurinated and irradiated DNA, respectively) generate nicks containing apurinic/apyrimidinic sites at their 3' termini and that such sites are not rapidly excised by the 3' leads to 5' activity of DNA polymerase I. However, endonuclease IV or VI apparently can remove such terminal apurinic/apyrimidinic sites as well as cleave on the 5' side of the unnicked sites. These results suggest roles for endonucleases III, IV, and VI in the repair of apurinic/apyrimidinic sites as well as pyrimidine dimer sites in DNA. Our results with T4 UV endonuclease suggest that the incision of irradiated DNA by T4 UV endonuclease involves both cleavage of the glycosylic bond at the 5' half of the pyrimidine dimer and cleavage of the phosphodiester bond originally linking the two nucleotides of the dimer. They also imply that the glycosylic bond is cleaved before the phosphodiester bond. PMID:6254032

  3. SLAC site design aesthetics

    SciTech Connect

    Hall, F.F.

    1985-10-01

    Stanford Linear Accelerator Center (SLAC) is a single mission laboratory dedicated to basic research in high energy particle physics. SLAC site also houses Stanford Synchrotron Radiation Laboratory (SSRL) which is a multi-mission laboratory for research using beams of ultraviolet light and low energy photons as emitted tangentially from SLAC colliding beam facilities. This paper discusses various aspects of SLAC site design aesthetics under the following headings: (1) imposed footprint of SLAC, (2) description of selected site, (3) use of earth cover for radiation and sight screens, (4) use of landscaping for cosmetic purposes, (5) use of exterior paint colors to soften SLAC impact on neighbors, (6) relocation of SLAC main entrance, (7) relocation of SLAC collider arcs and experimental hall, (8) parking lots and storage yards, and (9) land use zoning at SLAC.

  4. Site specific synthesis and polymerase bypass of oligonucleotides containing a 6-hydroxy-3,5,6,7-tetrahydro-9H-imidazo[1,2-a]purin-9-one base, an intermediate in the formation of 1,N2-etheno-2'-deoxyguanosine.

    PubMed

    Goodenough, Angela K; Kozekov, Ivan D; Zang, Hong; Choi, Jeong-Yun; Guengerich, F Peter; Harris, Thomas M; Rizzo, Carmelo J

    2005-11-01

    The reaction of DNA with certain bis-electrophiles such as chlorooxirane and chloroacetaldehyde produces etheno adducts. These lesions are highly miscoding, and some of the chemical agents that produce them have been shown to be carcinogenic in laboratory animals and in humans. An intermediate in the formation of 1,N2-ethenoguanine is 6-hydroxy-3,5,6,7-tetrahydro-9H-imidazo[1,2-a]purin-9-one (6-hydroxyethanoguanine), which undergoes conversion to the etheno adduct. The chemical properties and miscoding potential of the hydroxyethano adduct have not been previously studied. A synthesis of the hydroxyethano-adducted nucleoside was developed, and it was site specifically incorporated into oligonucleotides. This adduct had a half-life of between 24 and 48 h at neutral pH and 25 degrees C at the nucleoside and oligonucleotide levels. The miscoding potential of the hydroxyethano adduct was examined by primer extension reactions with the DNA polymerases Dpo4 and pol T7-, and the results were compared to the corresponding etheno-adducted oligonucleotide. Dpo4 preferentially incorporated dATP opposite the hydroxyethano adduct and dGTP opposite the etheno adduct; pol T7- preferentially incorporated dATP opposite the etheno adduct while dGTP and dATP were incorporated opposite the hydroxyethano adduct with nearly equal catalytic efficiencies. Collectively, these results indicate that the hydroxyethano adduct has a sufficient lifetime and miscoding properties to contribute to the mutagenic spectrum of chlorooxirane and related genotoxic species.

  5. Save Money on Site Design.

    ERIC Educational Resources Information Center

    Galbraith, Roger

    1999-01-01

    Examples are provided for ways of saving money during school athletic-site design, including plastic alternatives to concrete for site water drainage and cost-efficient considerations when earth moving and grading. Concluding comments address the importance of designers making timely decisions and monitoring progress to avoid unnecessary costs.…

  6. Instructional Design. [SITE 2001 Section].

    ERIC Educational Resources Information Center

    Boger, Stephanie, Ed.

    This document contains the papers on instructional design from the SITE (Society for Information Technology & Teacher Education) 2001 conference. Topics covered include: an adaptive e-framework for teacher training; assessing the integration of technology into the curriculum; promoting instructional planning; learning and using World Wide Web page…

  7. Instructional Design. [SITE 2002 Section].

    ERIC Educational Resources Information Center

    2002

    This document contains the following papers on instructional design from the SITE (Society for Information Technology & Teacher Education) 2002 conference: "Faculty Guidelines for the Video Development Process" (Ronald J. Abate and Kathleen Benghiat); "Evolution of an Online Graduate Course in Educational Multimedia" (Peter Albion); "PT3…

  8. Teaching Web Site Design: Science or Art?

    ERIC Educational Resources Information Center

    Taylor, Mark John; Salces, Fausto Sainz; Duffy, Sandra Patricia

    2005-01-01

    This paper discusses an approach to teaching web site design to higher education (HE) students that attempts to balance the artistic and scientific aspects of the web site design process. The design of IT systems has traditionally been viewed as a technical activity. However, unlike existing types of IT systems, an organization's web site is…

  9. Teaching Web Site Design: Science or Art?

    ERIC Educational Resources Information Center

    Taylor, Mark John; Salces, Fausto Sainz; Duffy, Sandra Patricia

    2005-01-01

    This paper discusses an approach to teaching web site design to higher education (HE) students that attempts to balance the artistic and scientific aspects of the web site design process. The design of IT systems has traditionally been viewed as a technical activity. However, unlike existing types of IT systems, an organization's web site is…

  10. Publicity through Better Web Site Design.

    ERIC Educational Resources Information Center

    Guenther, Kim

    1999-01-01

    Describes how to design Web sites that will reach the most people possible. Discusses publicity; Web search engine sites; adopting Web development standards; using metatags to effectively index a site; Web site addresses and naming conventions; writing for the Web; mass submissions; subject area knowledge; making Web rings; and additional ideas to…

  11. Physical association of pyrimidine dimer DNA glycosylase and apurinic/apyrimidinic DNA endonuclease essential for repair of ultraviolet-damaged DNA

    SciTech Connect

    Nakabeppu, Y.; Sekiguchi, M.

    1981-05-01

    T4 endonuclease, which is involved in repair of uv-damaged DNA, has been purified to apparent physical homogeneity. Incubation of uv-irradiated poly(dA).poly(dT) with the purified enzyme preparations resulted in production of alkali-labile apyrimidinic sites, followed by formation of nicks in the polymer. By performing a limited reaction with T4 endonuclease V at pH 8.5, irradiated polymer was converted to an intermediate form that carried a large number of alkali-labile sites but only a few nicks. The intermediate was used as substrate for the assay of apurinic/apyrimidinic DNA endonuclease activity. The two activities, a pyrimidine dimer DNA glycosylase and an apurinic/apyrimidinic DNA endonuclease, were copurified and found in enzyme preparations that contained only a 16,000-dalton polypeptide. These results strongly suggested that a DNA glycosylase specific for pyrimidine dimers and an apurinic/apyrimidinic DNA endonuclease reside in a single polypeptide chain coded by the denV gene of bacteriophage T4.

  12. GEOSTATISTICAL SAMPLING DESIGNS FOR HAZARDOUS WASTE SITES

    EPA Science Inventory

    This chapter discusses field sampling design for environmental sites and hazardous waste sites with respect to random variable sampling theory, Gy's sampling theory, and geostatistical (kriging) sampling theory. The literature often presents these sampling methods as an adversari...

  13. GEOSTATISTICAL SAMPLING DESIGNS FOR HAZARDOUS WASTE SITES

    EPA Science Inventory

    This chapter discusses field sampling design for environmental sites and hazardous waste sites with respect to random variable sampling theory, Gy's sampling theory, and geostatistical (kriging) sampling theory. The literature often presents these sampling methods as an adversari...

  14. Suppression of oxidative phosphorylation in mouse embryonic fibroblast cells deficient in apurinic/apyrimidinic endonuclease

    PubMed Central

    Suganya, Rangaswamy; Chakraborty, Anirban; Miriyala, Sumitra; Hazra, Tapas K.; Izumi, Tadahide

    2015-01-01

    The mammalian apurinic/apyrimidinic (AP) endonuclease 1 (APE1) is an essential DNA repair/gene regulatory protein. Decrease of APE1 in cells by inducible shRNA knockdown or by conditional gene knockout caused apoptosis. Here we succeeded in establishing a unique mouse embryonic fibroblast (MEF) line expressing APE1 at a level far lower than those achieved with shRNA knockdown. The cells, named MEFla (MEFlowAPE1), were hypersensitive to methyl methanesulfonate (MMS), and showed little activity for repairing AP-sites and MMS induced DNA damage. While these results were consistent with the essential role of APE1 in repair of AP sites, the MEFla cells grew normally and the basal activation of poly(ADP-ribose) polymerases in MEFla was lower than that in the wild-type MEF (MEFwt), indicating the low DNA damage stress in MEFla under the normal growth condition. Oxidative phosphorylation activity in MEFla was lower than in MEFwt, while the glycolysis rates in MEFla were higher than in MEFwt. In addition, we observed decreased intracellular oxidative stress in MEFla. These results suggest that cells with low APE1 reversibly suppress mitochondrial respiration and thereby reduce DNA damage stress and increases the cell viability. PMID:25645679

  15. Base excision repair apurinic/apyrimidinic endonucleases in apicomplexan parasite Toxoplasma gondii

    PubMed Central

    Onyango, David O.; Naguleswaran, Arunasalam; Delaplane, Sarah; Reed, April; Kelley, Mark R.; Georgiadis, Millie M.; Sullivan, William J.

    2011-01-01

    DNA repair is essential for cell viability and proliferation. In addition to reactive oxygen produced as a byproduct of their own metabolism, intracellular parasites also have to manage oxidative stress generated as a defense mechanism by the host. The spontaneous loss of DNA bases due to hydrolysis and oxidative DNA damage in intracellular parasites is great, but little is known about the type of DNA repair machineries that exist in these early-branching eukaryotes. However, it is clear processes similar to DNA base excision repair (BER) must exist to rectify spontaneous and host-mediated damage in Toxoplasma gondii. Here we report that Toxoplasma gondii, an opportunistic protozoan pathogen, possesses two apurinic/apyrimidinic (AP) endonucleases that function in DNA BER. We characterize the enzymatic activities of Toxoplasma exonuclease III (ExoIII, or Ape1) and endonuclease IV (EndoIV, or Apn1), designated TgAPE and TgAPN, respectively. Over-expression of TgAPN in Toxoplasma conferred protection from DNA damage, and viable knockouts of TgAPN were not obtainable. We generated an inducible TgAPN knockdown mutant using a ligand-controlled destabilization domain to establish that TgAPN is critical for Toxoplasma to recover from DNA damage. The importance of TgAPN and the fact that humans lack any observable APN family activity highlights TgAPN as a promising candidate for drug development to treat toxoplasmosis. PMID:21353648

  16. Characterization and genetic mapping of a mutation affecting apurinic endonuclease activity in Staphylococcus aureus.

    PubMed Central

    Tam, J E; Pattee, P A

    1986-01-01

    Protoplast fusion between the Rec- mutant RN981 (L. Wyman, R. V. Goering, and R. P. Novick, Genetics 76:681-702, 1974) of Staphylococcus aureus NCTC 8325 and a Rec+ NCTC 8325 derivative yielded Rec+ recombinants that exhibited the increased sensitivity to N-methyl-N'-nitro-N-nitrosoguanidine characteristic of RN981. Transformation analyses identified a specific mutation, designated ngr-374, that was responsible not only for N-methyl-N'-nitro-N-nitrosoguanidine sensitivity, but also sensitivity to methyl methanesulfonate, ethyl methanesulfonate, nitrous acid, and UV irradiation. However, ngr-374-carrying recombinants showed no significant increase in their sensitivity to mitomycin C or 4-nitroquinoline 1-oxide and were unaffected in recombination proficiency. In vitro assays showed that ngr-374-carrying strains had lower apurinic/apyrimidinic endonuclease activities than the wild type. The chromosomal locus occupied by ngr-374 was shown to exist in the gene order omega(Chr::Tn551)40-ngr-374-thrB106. PMID:2430940

  17. Human apurinic/apyrimidinic endonuclease 1 (APE1) has 3' RNA phosphatase and 3' exoribonuclease activities.

    PubMed

    Chohan, Manbir; Mackedenski, Sebastian; Li, Wai-Ming; Lee, Chow H

    2015-01-30

    Apurinic/apyrimidinic endonuclease 1 (APE1) is the predominant mammalian enzyme in DNA base excision repair pathway that cleaves the DNA backbone immediately 5' to abasic sites. In addition to its abasic endonuclease activity, APE1 has 3' phosphatase and 3'-5' exonuclease activities against DNA. We recently identified APE1 as an endoribonuclease that preferentially cleaves at UA, UG, and CA sites in single-stranded regions of RNAs and can regulate c-myc mRNA level and half-life in cells. APE1 can also endonucleolytically cleave abasic single-stranded RNA. Here, we show for the first time that the human APE1 has 3' RNA phosphatase and 3' exoribonuclease activities. Using three distinct RNA substrates, we show that APE1, but not RNase A, can remove the phosphoryl group from the 3' end of RNA decay products. Studies using various site-directed APE1 mutant proteins (H309N, H309S, D283N, N68A, D210N, Y171F, D308A, F266A, and D70A) suggest that the 3' RNA phosphatase activity shares the same active center as its other known nuclease activities. A number of APE1 variants previously identified in the human population, including the most common D148E variant, have greater than 80% reduction in the 3' RNA phosphatase activity. APE1 can remove a ribonucleotide from the 3' overhang of RNA decay product, but its 3'-5' exoribonuclease activity against unstructured poly(A), poly(C), and poly(U) RNAs is relatively weak. This study further underscores the significance of understanding the role of APE1 in RNA metabolism in vivo.

  18. AIAA designates Stennis as historic aerospace site

    NASA Technical Reports Server (NTRS)

    2008-01-01

    Picured (left to right) American Institute of Aeronautics and Astronautics (AIAA) representative David Throckmorton presents a plaque designating NASA's John C. Stennis Space Center as a historical aerospace site during an April 10 ceremony. Joining Throckmorton for the presentation were AIAA Greater New Orleans Chapter Chair Mark Hughes, Stennis Space Center Director Bob Cabana and Pratt & Whitney Rocketdyne Vice President John Plowden.

  19. Using Participatory Design to Improve Web Sites

    ERIC Educational Resources Information Center

    Nikolova-Houston, Tatiana

    2005-01-01

    In this article, the author, a doctoral candidate from the School of Information at the University of Texas-Austin, describes the experience gathered from her attempt to redesign her existing Web sites in order to supply online resources for Slavic and Byzantine studies. The use of participatory design, which involves the users in creating the…

  20. AIAA designates Stennis as historic aerospace site

    NASA Image and Video Library

    2008-04-10

    Picured (left to right) American Institute of Aeronautics and Astronautics (AIAA) representative David Throckmorton presents a plaque designating NASA's John C. Stennis Space Center as a historical aerospace site during an April 10 ceremony. Joining Throckmorton for the presentation were AIAA Greater New Orleans Chapter Chair Mark Hughes, Stennis Space Center Director Bob Cabana and Pratt & Whitney Rocketdyne Vice President John Plowden.

  1. Using Participatory Design to Improve Web Sites

    ERIC Educational Resources Information Center

    Nikolova-Houston, Tatiana

    2005-01-01

    In this article, the author, a doctoral candidate from the School of Information at the University of Texas-Austin, describes the experience gathered from her attempt to redesign her existing Web sites in order to supply online resources for Slavic and Byzantine studies. The use of participatory design, which involves the users in creating the…

  2. Access to Space Interactive Design Web Site

    NASA Technical Reports Server (NTRS)

    Leon, John; Cutlip, William; Hametz, Mark

    2000-01-01

    The Access To Space (ATS) Group at NASA's Goddard Space Flight Center (GSFC) supports the science and technology community at GSFC by facilitating frequent and affordable opportunities for access to space. Through partnerships established with access mode suppliers, the ATS Group has developed an interactive Mission Design web site. The ATS web site provides both the information and the tools necessary to assist mission planners in selecting and planning their ride to space. This includes the evaluation of single payloads vs. ride-sharing opportunities to reduce the cost of access to space. Features of this site include the following: (1) Mission Database. Our mission database contains a listing of missions ranging from proposed missions to manifested. Missions can be entered by our user community through data input tools. Data is then accessed by users through various search engines: orbit parameters, ride-share opportunities, spacecraft parameters, other mission notes, launch vehicle, and contact information. (2) Launch Vehicle Toolboxes. The launch vehicle toolboxes provide the user a full range of information on vehicle classes and individual configurations. Topics include: general information, environments, performance, payload interface, available volume, and launch sites.

  3. Conceptual design report for site drainage control

    SciTech Connect

    Hunter, M.R.

    1996-07-01

    The Mound Plant (Mound), located in Miamisburg, Ohio, is a Department of Energy (DOE) development and production facility performing support work for DOE`s weapons and energy-related programs. EG&G Mound Applied Technologies, Inc. (EG&G) is the Operating Contractor (OC) for this Government-Owned, Contractor-Operated (GOCO) facility. The work performed at Mound emphasizes nuclear energy and explosives technology. Mound is currently implementing an Environmental, Safety & Health (ES&H) Upgrades Program designed to protect its employees, the public, and the environment from adverse effects caused by facility activities. The first project of this multiphase program is now in the final stages of construction, and the second project is currently under design. Four additional projects, one of which is presented in this report, are in the conceptual design stage. At Mound, 22 soil zones have become contaminated with radioactive material. These zones cover approximately 20 percent of the total area of developed property at the site. During a storm event, the rainwater washes contaminated soil from these zones into the storm sewer system. These radioactive contaminants may then be discharged along with the stormwater into the Great Miami River via the Miami Erie Canal. This conceptual design report (CDR), Site Drainage Control, the fourth project in the ES&H program, describes a project that will provide improvements and much needed repairs to inadequate and deteriorating portions of the storm drainage system on the developed property. The project also will provide a stormwater retention facility capable of storing the stormwater runoff, from the developed property, resulting from a 100-year storm event. These improvements will permit the effective control and monitoring of stormwater to prevent the spread of radioactive contaminants from contaminated soil zones and will provide a means to collect and contain accidental spills of hazardous substances.

  4. 10 CFR 862.7 - Designation of sites.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 10 Energy 4 2010-01-01 2010-01-01 false Designation of sites. 862.7 Section 862.7 Energy DEPARTMENT OF ENERGY RESTRICTIONS ON AIRCRAFT LANDING AND AIR DELIVERY AT DEPARTMENT OF ENERGY NUCLEAR SITES § 862.7 Designation of sites. (a) DOE shall designate sites covered by this part as deemed...

  5. Repository surface design site layout analysis

    SciTech Connect

    Montalvo, H.R.

    1998-02-27

    The purpose of this analysis is to establish the arrangement of the Yucca Mountain Repository surface facilities and features near the North Portal. The analysis updates and expands the North Portal area site layout concept presented in the ACD, including changes to reflect the resizing of the Waste Handling Building (WHB), Waste Treatment Building (WTB), Carrier Preparation Building (CPB), and site parking areas; the addition of the Carrier Washdown Buildings (CWBs); the elimination of the Cask Maintenance Facility (CMF); and the development of a concept for site grading and flood control. The analysis also establishes the layout of the surface features (e.g., roads and utilities) that connect all the repository surface areas (North Portal Operations Area, South Portal Development Operations Area, Emplacement Shaft Surface Operations Area, and Development Shaft Surface Operations Area) and locates an area for a potential lag storage facility. Details of South Portal and shaft layouts will be covered in separate design analyses. The objective of this analysis is to provide a suitable level of design for the Viability Assessment (VA). The analysis was revised to incorporate additional material developed since the issuance of Revision 01. This material includes safeguards and security input, utility system input (size and location of fire water tanks and pump houses, potable water and sanitary sewage rates, size of wastewater evaporation pond, size and location of the utility building, size of the bulk fuel storage tank, and size and location of other exterior process equipment), main electrical substation information, redundancy of water supply and storage for the fire support system, and additional information on the storm water retention pond.

  6. Repository Surface Design Site Layout Analysis

    SciTech Connect

    Montalvo, H.R.

    1998-02-27

    The purpose of this analysis is to establish the arrangement of the Repository surface facilities and features near the North Portal. The analysis updates and expands the North Portal area site layout concept presented in the ACD (Reference 5.5), including changes to reflect the resizing of the Waste Handling Building (WHB), Waste Treatment Building (WTB), Carrier Preparation Building (CPB), and site parking areas; the addition of the Carrier Washdown Buildings (CWBs); the elimination of the Cask Maintenance Facility (CMF); and the development of a concept for site grading and flood control. The analysis also establishes the layout of the surface features (e.g., roads and utilities) that connect all the repository surface areas (North Portal Operations Area, South Portal Development Operations Area, Emplacement Shaft Surface Operations Area, and Development Shaft Surface Operations Area) and locates an area for a potential lag storage facility. Details of South Portal and shaft layouts will be covered in separate design analyses. The objective of this analysis is to provide a suitable level of design for the Viability Assessment (VA). The analysis was revised to incorporate additional material developed since the issuance of Revision 01. This material includes safeguards and security input, utility system input (size and location of fire water tanks and pump houses, potable water and sanitary sewage rates, size of wastewater evaporation pond, size and location of the utility building, size of the bulk fuel storage tank, and size and location of other exterior process equipment), main electrical substation information, redundancy of water supply and storage for the fire support system, and additional information on the storm water retention pond.

  7. Characterization of DNA substrate specificities of apurinic/apyrimidinic endonucleases from Mycobacterium tuberculosis.

    PubMed

    Abeldenov, Sailau; Talhaoui, Ibtissam; Zharkov, Dmitry O; Ishchenko, Alexander A; Ramanculov, Erlan; Saparbaev, Murat; Khassenov, Bekbolat

    2015-09-01

    Apurinic/apyrimidinic (AP) endonucleases are key enzymes involved in the repair of abasic sites and DNA strand breaks. Pathogenic bacteria Mycobacterium tuberculosis contains two AP endonucleases: MtbXthA and MtbNfo members of the exonuclease III and endonuclease IV families, which are exemplified by Escherichia coli Xth and Nfo, respectively. It has been shown that both MtbXthA and MtbNfo contain AP endonuclease and 3'→5' exonuclease activities. However, it remains unclear whether these enzymes hold 3'-repair phosphodiesterase and nucleotide incision repair (NIR) activities. Here, we report that both mycobacterial enzymes have 3'-repair phosphodiesterase and 3'-phosphatase, and MtbNfo contains in addition a very weak NIR activity. Interestingly, depending on pH, both enzymes require different concentrations of divalent cations: 0.5mM MnCl2 at pH 7.6 and 10 mM at pH 6.5. MtbXthA requires a low ionic strength and 37 °C, while MtbNfo requires high ionic strength (200 mM KCl) and has a temperature optimum at 60 °C. Point mutation analysis showed that D180 and N182 in MtbXthA and H206 and E129 in MtbNfo are critical for enzymes activities. The steady-state kinetic parameters indicate that MtbXthA removes 3'-blocking sugar-phosphate and 3'-phosphate moieties at DNA strand breaks with an extremely high efficiency (kcat/KM=440 and 1280 μM(-1)∙min(-1), respectively), while MtbNfo exhibits much lower 3'-repair activities (kcat/KM=0.26 and 0.65 μM(-1)∙min(-1), respectively). Surprisingly, both MtbXthA and MtbNfo exhibited very weak AP site cleavage activities, with kinetic parameters 100- and 300-fold lower, respectively, as compared with the results reported previously. Expression of MtbXthA and MtbNfo reduced the sensitivity of AP endonuclease-deficient E. coli xth nfo strain to methylmethanesulfonate and H2O2 to various degrees. Taken together, these data establish the DNA substrate specificity of M. tuberculosis AP endonucleases and suggest their possible role

  8. A Design Analysis Model for Developing World Wide Web Sites.

    ERIC Educational Resources Information Center

    Ma, Yan

    2002-01-01

    Examines the relationship between and among designers, text, and users of the Galter Health Sciences Library Web site at Northwestern University by applying reader-response criticism. Highlights include Web site design; comparison of designers' intentions with the actual organization of knowledge on the Web site; and compares designer's intentions…

  9. Lucanthone and its derivative hycanthone inhibit apurinic endonuclease-1 (APE1) by direct protein binding

    SciTech Connect

    Naidu, M.; Naidu, M.; Agarwal, R.; Pena, L.A.; Cunha, L.; Mezei, M.; Shen, M.; Wilson, D.M.; Liu, Y.; Sanchez, Z.; Chaudhary, P.; Wilson, S.H.; Waring, M.J.

    2011-09-15

    Lucanthone and hycanthone are thioxanthenone DNA intercalators used in the 1980s as antitumor agents. Lucanthone is in Phase I clinical trial, whereas hycanthone was pulled out of Phase II trials. Their potential mechanism of action includes DNA intercalation, inhibition of nucleic acid biosyntheses, and inhibition of enzymes like topoisomerases and the dual function base excision repair enzyme apurinic endonuclease 1 (APE1). Lucanthone inhibits the endonuclease activity of APE1, without affecting its redox activity. Our goal was to decipher the precise mechanism of APE1 inhibition as a prerequisite towards development of improved therapeutics that can counteract higher APE1 activity often seen in tumors. The IC{sub 50} values for inhibition of APE1 incision of depurinated plasmid DNA by lucanthone and hycanthone were 5 {mu}M and 80 nM, respectively. The K{sub D} values (affinity constants) for APE1, as determined by BIACORE binding studies, were 89 nM for lucanthone/10 nM for hycanthone. APE1 structures reveal a hydrophobic pocket where hydrophobic small molecules like thioxanthenones can bind, and our modeling studies confirmed such docking. Circular dichroism spectra uncovered change in the helical structure of APE1 in the presence of lucanthone/hycanthone, and notably, this effect was decreased (Phe266Ala or Phe266Cys or Trp280Leu) or abolished (Phe266Ala/Trp280Ala) when hydrophobic site mutants were employed. Reduced inhibition by lucanthone of the diminished endonuclease activity of hydrophobic mutant proteins (as compared to wild type APE1) supports that binding of lucanthone to the hydrophobic pocket dictates APE1 inhibition. The DNA binding capacity of APE1 was marginally inhibited by lucanthone, and not at all by hycanthone, supporting our hypothesis that thioxanthenones inhibit APE1, predominantly, by direct interaction. Finally, lucanthone-induced degradation was drastically reduced in the presence of short and long lived free radical scavengers, e

  10. Cloning and Characterization of a Wheat Homologue of Apurinic/Apyrimidinic Endonuclease Ape1L

    PubMed Central

    Grin, Inga R.; Zharkov, Dmitry O.; Ishenko, Alexander A.; Tudek, Barbara; Bissenbaev, Amangeldy K.; Saparbaev, Murat

    2014-01-01

    Background Apurinic/apyrimidinic (AP) endonucleases are key DNA repair enzymes involved in the base excision repair (BER) pathway. In BER, an AP endonuclease cleaves DNA at AP sites and 3′-blocking moieties generated by DNA glycosylases and/or oxidative damage. A Triticum aestivum cDNA encoding for a putative homologue of ExoIII family AP endonucleases which includes E. coli Xth, human APE1 and Arabidopsis thaliana AtApe1L has been isolated and its protein product purified and characterized. Methodology/Principal Findings We report that the putative wheat AP endonuclease, referred here as TaApe1L, contains AP endonuclease, 3′-repair phosphodiesterase, 3′-phosphatase and 3′→5′ exonuclease activities. Surprisingly, in contrast to bacterial and human AP endonucleases, addition of Mg2+ and Ca2+ (5–10 mM) to the reaction mixture inhibited TaApe1L whereas the presence of Mn2+, Co2+ and Fe2+ cations (0.1–1.0 mM) strongly stimulated all its DNA repair activities. Optimization of the reaction conditions revealed that the wheat enzyme requires low divalent cation concentration (0.1 mM), mildly acidic pH (6–7), low ionic strength (20 mM KCl) and has a temperature optimum at around 20°C. The steady-state kinetic parameters of enzymatic reactions indicate that TaApe1L removes 3′-blocking sugar-phosphate and 3′-phosphate groups with good efficiency (kcat/KM = 630 and 485 μM−1·min−1, respectively) but possesses a very weak AP endonuclease activity as compared to the human homologue, APE1. Conclusions/Significance Taken together, these data establish the DNA substrate specificity of the wheat AP endonuclease and suggest its possible role in the repair of DNA damage generated by endogenous and environmental factors. PMID:24667595

  11. Construction of a Full-Atomic Mechanistic Model of Human Apurinic/Apyrimidinic Endonuclease APE1 for Virtual Screening of Novel Inhibitors.

    PubMed

    Khaliullin, I G; Nilov, D K; Shapovalova, I V; Svedas, V K

    2012-04-01

    A full-atomic molecular model of human apurinic/apyrimidinic endonuclease APE1, an important enzyme in the DNA repair system, has been constructed. The research consisted of hybrid quantum mechanics/molecular mechanics modeling of the enzyme-substrate interactions, as well as calculations of the ionization states of the amino acid residues of the active site of the enzyme. The choice of the APE1 mechanism with an Asp210 residue as a proton acceptor was validated by means of a generalization of modeling and experimental data. Interactions were revealed in the active site that are of greatest significance for binding the substrate and potential APE1 inhibitors (potential co-drugs of interest in the chemo- and radiotherapy of oncological diseases).

  12. 40 CFR 228.4 - Procedures for designation of sites.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... designation or evaluation studies on a typical site or sites in that area. In such cases, sufficient studies... environmental studies of each site, regions adjacent to the site, and on historical knowledge of the impact of... studies for the evaluation and potential selection of dumping sites will be conducted in accordance...

  13. Synthetic lethal targeting of DNA double strand break repair deficient cells by human apurinic/apyrimidinic endonuclease (APE1) inhibitors

    PubMed Central

    Sultana, Rebeka; McNeill, Daniel R.; Abbotts, Rachel; Mohammed, Mohammed Z.; Zdzienicka, Małgorzata Z.; Qutob, Haitham; Seedhouse, Claire; Laughton, Charles A.; Fischer, Peter M.; Patel, Poulam M.; Wilson, David M.; Madhusudan, Srinivasan

    2013-01-01

    An apurinic/apyrimidinic (AP) site is an obligatory cytotoxic intermediate in DNA Base Excision Repair (BER) that is processed by human AP endonuclease 1 (APE1). APE1 is essential for BER and an emerging drug target in cancer. We have isolated novel small molecule inhibitors of APE1. In the current study we have investigated the ability of APE1 inhibitors to induce synthetic lethality in a panel of DNA double strand break (DSB) repair deficient and proficient cells; a) Chinese hamster (CH) cells: BRCA2 deficient (V-C8), ATM deficient (V-E5), wild type (V79) and BRCA2 revertant (V-C8(Rev1)). b) Human cancer cells: BRCA1 deficient (MDA-MB-436), BRCA1 proficient (MCF-7), BRCA2 deficient (CAPAN-1 and HeLa SilenciX cells), BRCA2 proficient (PANC1 and control SilenciX cells). We also tested synthetic lethality (SL) in CH ovary cells expressing a dominant–negative form of APE1 (E8 cells) using ATM inhibitors and DNA-PKcs inhibitors (DSB inhibitors). APE1 inhibitors are synthetically lethal in BRCA and ATM deficient cells. APE1 inhibition resulted in accumulation of DNA DSBs and G2/M cell cycle arrest. Synthetic lethality was also demonstrated in CH cells expressing a dominant–negative form of APE1 treated with ATM or DNA-PKcs inhibitors. We conclude that APE1 is a promising synthetic lethality target in cancer. PMID:22377908

  14. Web Site Design Benchmarking within Industry Groups.

    ERIC Educational Resources Information Center

    Kim, Sung-Eon; Shaw, Thomas; Schneider, Helmut

    2003-01-01

    Discussion of electronic commerce focuses on Web site evaluation criteria and applies them to different industry groups in Korea. Defines six categories of Web site evaluation criteria: business function, corporate credibility, contents reliability, Web site attractiveness, systematic structure, and navigation; and discusses differences between…

  15. Web Site Design Benchmarking within Industry Groups.

    ERIC Educational Resources Information Center

    Kim, Sung-Eon; Shaw, Thomas; Schneider, Helmut

    2003-01-01

    Discussion of electronic commerce focuses on Web site evaluation criteria and applies them to different industry groups in Korea. Defines six categories of Web site evaluation criteria: business function, corporate credibility, contents reliability, Web site attractiveness, systematic structure, and navigation; and discusses differences between…

  16. Design criteria for stabilization of inactive uranium mill tailings sites

    SciTech Connect

    1984-06-01

    The site-specific Remedial Action Plan (RAP), which includes the Site Conceputal Design (SCD); the Processing Site Characterization Report (PSCR); the Disposal Site Characterization Report (DSCR), (if required); and the project Site Design Criteria (SDC), as a set, have been prepared by the Technical Assistance Contractor (TAC) at the direction of the Department of Energy (DOE), and provide the Remedial Action Contractor (RAC) with sufficient information to complete the remedial action engineering designs. This Site Design Criteria (SDC) provides the basis or guidelines for the RAC to prepare the final design documentation for the Uranium Mill Tailings Remedial Action (UMTRA) Project sites. This document provides a set of operating procedures, formats for drawings, specifications, calculations, schedules and cost estimates, and minimum design constraints.

  17. Brownfields Recommendations for Sustainable Site Design — Green Landscape Plan

    EPA Pesticide Factsheets

    The assessment of conditions contained in this report focuses on site-specific environmental and soil conditions that might affect recommendations related to sustainable landscaping and site design, stormwater management, and stormwater reuse.

  18. Determination of apurinic/apyrimidinic lesions in DNA with high-performance liquid chromatography and tandem mass spectrometry.

    PubMed

    Roberts, Kenneth P; Sobrino, Justin A; Payton, Julie; Mason, Lavinnia B; Turesky, Robert J

    2006-02-01

    A new method has been developed to accurately measure apurinic and apyrimidinic (AP) DNA damage sites, which are lesions in DNA formed by loss of a nucleobase from oxidative stress or carcinogen adducts. If AP sites are left unrepaired (or if improperly repaired), these sites can lead to DNA mutations that may ultimately result in the formation of cancer. Hence, detection of AP sites may provide a useful indicator of exposure and susceptibility to chemical carcinogens and oxidative stress. AP detection is currently accomplished by immunodetection methods using an aldehyde reactive probe [Nakamura, J., Walker, V. E., Upton, P. B., Chiang, S.-Y., Kow, Y. W., and Swenberg, J. A. (1998) Cancer Res. 58, 222-225; Atamna, H., Cheung, I., and Ames, B. N. (2000) Proc. Natl. Acad. Sci. U.S.A. 97, 686-691]; however, these approaches lack the specificity required for unequivocal identification of the AP site. Therefore, we have developed an accurate method based on mass spectrometry detection of AP sites from AP DNA that have been prelabeled with O-4-nitrobenzylhydroxylamine (NBHA). Once labeled and once the excess labeling agent has been removed, enzymatic digestion of DNA to monomeric subunits can be accomplished, followed by isolation and detection with high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS). Optimization and validation of the experimental procedures and detection limits have been established using a model DNA oligomer (11-mer) containing uracil. Enzymatic removal of uracil with uracil glycosylase generates well-defined AP sites in both single- and double-stranded DNA. The addition of NBHA labels the AP site in the oligomer, creating a labeled 11-mer. HPLC-ESI-MS/MS in the negative ionization mode was used to monitor and confirm binding of NBHA to the AP oligomer. The NBHA-tagged oligomer underwent endo- and exonuclease digestion to the 5'-deoxyribose monophosphate (5'-dRp) level, thereby releasing

  19. Amino-acid site variability among natural and designed proteins.

    PubMed

    Jackson, Eleisha L; Ollikainen, Noah; Covert, Arthur W; Kortemme, Tanja; Wilke, Claus O

    2013-01-01

    Computational protein design attempts to create protein sequences that fold stably into pre-specified structures. Here we compare alignments of designed proteins to alignments of natural proteins and assess how closely designed sequences recapitulate patterns of sequence variation found in natural protein sequences. We design proteins using RosettaDesign, and we evaluate both fixed-backbone designs and variable-backbone designs with different amounts of backbone flexibility. We find that proteins designed with a fixed backbone tend to underestimate the amount of site variability observed in natural proteins while proteins designed with an intermediate amount of backbone flexibility result in more realistic site variability. Further, the correlation between solvent exposure and site variability in designed proteins is lower than that in natural proteins. This finding suggests that site variability is too uniform across different solvent exposure states (i.e., buried residues are too variable or exposed residues too conserved). When comparing the amino acid frequencies in the designed proteins with those in natural proteins we find that in the designed proteins hydrophobic residues are underrepresented in the core. From these results we conclude that intermediate backbone flexibility during design results in more accurate protein design and that either scoring functions or backbone sampling methods require further improvement to accurately replicate structural constraints on site variability.

  20. Amino-acid site variability among natural and designed proteins

    PubMed Central

    Jackson, Eleisha L.; Ollikainen, Noah; Covert, Arthur W.; Kortemme, Tanja

    2013-01-01

    Computational protein design attempts to create protein sequences that fold stably into pre-specified structures. Here we compare alignments of designed proteins to alignments of natural proteins and assess how closely designed sequences recapitulate patterns of sequence variation found in natural protein sequences. We design proteins using RosettaDesign, and we evaluate both fixed-backbone designs and variable-backbone designs with different amounts of backbone flexibility. We find that proteins designed with a fixed backbone tend to underestimate the amount of site variability observed in natural proteins while proteins designed with an intermediate amount of backbone flexibility result in more realistic site variability. Further, the correlation between solvent exposure and site variability in designed proteins is lower than that in natural proteins. This finding suggests that site variability is too uniform across different solvent exposure states (i.e., buried residues are too variable or exposed residues too conserved). When comparing the amino acid frequencies in the designed proteins with those in natural proteins we find that in the designed proteins hydrophobic residues are underrepresented in the core. From these results we conclude that intermediate backbone flexibility during design results in more accurate protein design and that either scoring functions or backbone sampling methods require further improvement to accurately replicate structural constraints on site variability. PMID:24255821

  1. Specifications for the Design of the School Site.

    ERIC Educational Resources Information Center

    Brubaker, C. William

    1986-01-01

    The school site design is now seen as part of community planning. Recreational, cultural, social, and educational facilities for adults are included in educational specifications. Three illustrations and two photographs demonstrated attractive school sites. Site area requirements for a new high school in Santa Fe, New Mexico are summarized. (MLF)

  2. Specifications for the Design of the School Site.

    ERIC Educational Resources Information Center

    Brubaker, C. William

    1986-01-01

    The school site design is now seen as part of community planning. Recreational, cultural, social, and educational facilities for adults are included in educational specifications. Three illustrations and two photographs demonstrated attractive school sites. Site area requirements for a new high school in Santa Fe, New Mexico are summarized. (MLF)

  3. Culture differences in design of portal sites.

    PubMed

    Fang, Xiaowen; Rau, Pei-Luen Patrick

    2003-01-15

    Two experiments were carried out to examine the effects of cultural differences between the Chinese and the US people on the perceived usability and search performance of World Wide Web (WWW) portal sites. Chinese users in Taiwan and US users in Chicago were recruited to perform searching tasks on two versions of Yahoo! portal site: the standard Yahoo! and Yahoo! Chinese. The layout of Yahoo! Chinese is the same as the layout of Yahoo!, and categories on Yahoo! Chinese have been translated from its US counterpart. A special browser was programmed to record all the keystroke data and participants were asked to fill out a satisfaction questionnaire after finishing the tasks. Significant differences of satisfaction and steps to perform some tasks were found between the two groups. The experiment results also provided more detailed insights into the cultural differences between the Chinese and the US users.

  4. HVDC power transmission electrode siting and design

    SciTech Connect

    Holt, R.J.; Dabkowski, J.; Hauth, R.L.

    1997-04-01

    This research strives to shed light on the feasibility and practicality of using deep earth electrodes to permit their use for extended periods without adverse consequences. This report begins with a review of the fundamentals associated with current conduction in earth, including the various techniques available for measuring the earth`s electrical properties. The sources of existing data are discussed and some specific data for selected regions of the U.S. and Canada are reviewed as examples. Electrode technology and design issues are reviewed and recent experience gained by New England Power and Hydro-Quebec is discussed. The issues associated with direct current flowing in underground pipelines (and other facilities) are described and the present-day mitigation measures are evaluated. Suggestions are made for further R&D in the coordination of cathodic protection systems, an area that has evolved as an empirical, trial- and-error art more than a science.

  5. Design control for the Savannah River Site Consolidated Incineration Facility

    SciTech Connect

    Walker, R.E.; Rider, R.L. )

    1991-01-01

    The initiation, development, and control of the design for the Consolidated Incineration Facility at the US Department of Energy Savannah River Site has been, from the inception, a precisely and formally controlled process. A plan was developed and implemented to ensure output properly aligned with approved design criteria and conformed to applicable regulations throughout the design process. The key element of design control was the technical baseline which established the benchmark against which all changes to the design was evaluated. During the conceptual design phase of the project, design criteria were written to reflect the project objectives and functional requirements. Governmental regulations were reviewed to determine permitting and licensing actions required. Hazards assessments were performed to establish design classifications. The resulting design criteria, permitting requirements, and facility classifications were incorporated into the design plan which provided the basis for subsequent design activities. As the project proceeded through the various design phases, design control was maintained according to the design plan. Review of all design products was performed by the project team routinely. Formal independent design reviews were accomplished prior to releasing the design for construction. Alignment between criteria and design output was verified periodically throughout the design process. A formal design change control board was invoked to effect design changes impacting technical baselines. All changes to design initiated following issue for construction also were subject to procedural control.

  6. Design control for the Savannah River Site Consolidated Incineration Facility

    SciTech Connect

    Walker, R.E.; Rider, R.L.

    1991-12-31

    The initiation, development, and control of the design for the Consolidated Incineration Facility at the US Department of Energy Savannah River Site has been, from the inception, a precisely and formally controlled process. A plan was developed and implemented to ensure output properly aligned with approved design criteria and conformed to applicable regulations throughout the design process. The key element of design control was the technical baseline which established the benchmark against which all changes to the design was evaluated. During the conceptual design phase of the project, design criteria were written to reflect the project objectives and functional requirements. Governmental regulations were reviewed to determine permitting and licensing actions required. Hazards assessments were performed to establish design classifications. The resulting design criteria, permitting requirements, and facility classifications were incorporated into the design plan which provided the basis for subsequent design activities. As the project proceeded through the various design phases, design control was maintained according to the design plan. Review of all design products was performed by the project team routinely. Formal independent design reviews were accomplished prior to releasing the design for construction. Alignment between criteria and design output was verified periodically throughout the design process. A formal design change control board was invoked to effect design changes impacting technical baselines. All changes to design initiated following issue for construction also were subject to procedural control.

  7. Phase I privatization, site development, and roads design requirements document

    SciTech Connect

    Parazin, R.J.

    1996-09-30

    To prepare for the privatization contractor development of their assigned sites, roads and rail system must be extended from the existing area network systems. Various road and rail modification alternatives were investigated through an engineering study, @C-SD-TWR-ES- 004, with the preferred transportation corridors identified. Various Site development alternatives were also investigated, WHC-SD-TWR-ES-003. The preferred alternative, as discussed herein, will: 1. Establish boundaries and set monuments for the two PC sites. All work within each designated site will be by the PC. 2. Assure that the systems to serve each site - Roads and Rail System Modifications, Raw and Potable Water Service, Liquid Effluent Transfer Systems and Electric Power- are integrated with each other to best serve the sites. 3. Identify the corridors for Low Activity Waste (LAW) feed lines from the existing AP Tank Farm to the PC sites. Site development project will prepare these corridors for construction ofthe feed lines by the PC. Ifrequired, these corridors would include room for the optional High Level Waste (HLW) feed line. 4. Perform necessary site earthwork and grading outside of the PC sites. 1.3 DOCUMENT OVERVIEW This DRD will list the documents that will form the basis for design and construction of the roads and rail modifications and site development.

  8. SiteOut: An Online Tool to Design Binding Site-Free DNA Sequences

    PubMed Central

    Scholes, Clarissa; Wunderlich, Zeba; DePace, Angela H.

    2016-01-01

    DNA-binding proteins control many fundamental biological processes such as transcription, recombination and replication. A major goal is to decipher the role that DNA sequence plays in orchestrating the binding and activity of such regulatory proteins. To address this goal, it is useful to rationally design DNA sequences with desired numbers, affinities and arrangements of protein binding sites. However, removing binding sites from DNA is computationally non-trivial since one risks creating new sites in the process of deleting or moving others. Here we present an online binding site removal tool, SiteOut, that enables users to design arbitrary DNA sequences that entirely lack binding sites for factors of interest. SiteOut can also be used to delete sites from a specific sequence, or to introduce site-free spacers between functional sequences without creating new sites at the junctions. In combination with commercial DNA synthesis services, SiteOut provides a powerful and flexible platform for synthetic projects that interrogate regulatory DNA. Here we describe the algorithm and illustrate the ways in which SiteOut can be used; it is publicly available at https://depace.med.harvard.edu/siteout/. PMID:26987123

  9. SiteOut: An Online Tool to Design Binding Site-Free DNA Sequences.

    PubMed

    Estrada, Javier; Ruiz-Herrero, Teresa; Scholes, Clarissa; Wunderlich, Zeba; DePace, Angela H

    2016-01-01

    DNA-binding proteins control many fundamental biological processes such as transcription, recombination and replication. A major goal is to decipher the role that DNA sequence plays in orchestrating the binding and activity of such regulatory proteins. To address this goal, it is useful to rationally design DNA sequences with desired numbers, affinities and arrangements of protein binding sites. However, removing binding sites from DNA is computationally non-trivial since one risks creating new sites in the process of deleting or moving others. Here we present an online binding site removal tool, SiteOut, that enables users to design arbitrary DNA sequences that entirely lack binding sites for factors of interest. SiteOut can also be used to delete sites from a specific sequence, or to introduce site-free spacers between functional sequences without creating new sites at the junctions. In combination with commercial DNA synthesis services, SiteOut provides a powerful and flexible platform for synthetic projects that interrogate regulatory DNA. Here we describe the algorithm and illustrate the ways in which SiteOut can be used; it is publicly available at https://depace.med.harvard.edu/siteout/.

  10. Site Analysis--Rudimentary Concept in Residential Design

    ERIC Educational Resources Information Center

    Nystrom, Dennis C.

    1969-01-01

    This article on site analysis is concerned with the external factors affecting the dwelling and surrounding area when planning a residential design. Schematic drawings as well as a list of important data to be considered when planning are included. (GR)

  11. OSU TOMF Program Site Selection and Preliminary Concept Design Report

    SciTech Connect

    Spadling, Steve

    2012-05-10

    The purpose of this report is to confirm the programmatic requirements for the new facilities, identify the most appropriate project site, and develop preliminary site and building concepts that successfully address the overall project goals and site issues. These new facilities will be designed to accommodate the staff, drivers and maintenance requirements for the future mixed fleet of passenger vehicles, Transit Style Buses and School Buses.

  12. 40 CFR 228.4 - Procedures for designation of sites.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... DUMPING CRITERIA FOR THE MANAGEMENT OF DISPOSAL SITES FOR OCEAN DUMPING § 228.4 Procedures for designation... permits. Areas where ocean dumping is permitted subject to the specific conditions of individual special... studies for the evaluation and potential selection of dumping sites will be conducted in accordance...

  13. Designing Search: Effective Search Interfaces for Academic Library Web Sites

    ERIC Educational Resources Information Center

    Teague-Rector, Susan; Ghaphery, Jimmy

    2008-01-01

    Academic libraries customize, support, and provide access to myriad information systems, each with complex graphical user interfaces. The number of possible information entry points on an academic library Web site is both daunting to the end-user and consistently challenging to library Web site designers. Faced with the challenges inherent in…

  14. Designing Search: Effective Search Interfaces for Academic Library Web Sites

    ERIC Educational Resources Information Center

    Teague-Rector, Susan; Ghaphery, Jimmy

    2008-01-01

    Academic libraries customize, support, and provide access to myriad information systems, each with complex graphical user interfaces. The number of possible information entry points on an academic library Web site is both daunting to the end-user and consistently challenging to library Web site designers. Faced with the challenges inherent in…

  15. Molecular Recognition of DNA Damage Sites by Apurinic/Apyrimidinic Endonucleases

    SciTech Connect

    Braun, W. A.

    2005-07-28

    The DNA repair/redox factor AP endonuclease 1 (APE1) is a multifunctional protein which is known to to be essential for DNA repair activity in human cells. Structural/functional analyses of the APE activity is thus been an important research field to assess cellular defense mechanisms against ionizing radiation.

  16. Synthesis and proton-NMR studies of oligonucleotides containing an apurinic (AP) site

    SciTech Connect

    Raap, J.; Dreef, C.E.; van der Marel, G.A.; van Boom, J.H.; Hilbers, C.W. )

    1987-10-01

    In order to elucidate the conformational properties of base-deleted oligodeoxyribonucleotides, the molecules d-CpS(pCpG)n (n = 1,2; S = sugar) were synthesized by the phosphotriester method and characterized by 1H-NMR spectroscopy. Complete assignment of all non-exchangeable proton resonances of both compounds was obtained by 1D- and 2D-NMR techniques. In combination with computer simulation, these spectra yielded proton-proton and proton-phosphorus coupling constants of high accuracy. These data provide valuable information about the sugar and the backbone conformation. It appears that d-Cp1Sp2Cp3G4 does not form a duplex under any of the conditions studied. On the contrary, the base-deleted hexamer d-Cp1Sp2Cp3Gp4Cp5G6 occurs as a right-handed' staggered' DNA duplex at 280 K: the core of this duplex is formed by the residues C(3)-G(6); two 'dangling' residues C(1) and S(2) are located at the two 5'-ends of the duplex. The assignment of the corresponding imino proton resonances for (d-CpS(pCpG)2)2 was based on their thermal behavior: the line broadening of these resonances was studied as a function of temperature. The chemical shift and the number of imino proton resonances accord well with the number and type of Watson-Crick base pairs which can be formed in the staggered duplex described above. Thermodynamic parameters of duplex formation were obtained from an analysis of the chemical shift versus temperature profiles of aromatic base and H-1' protons. It is suggested that the cytosine ring of C(1) stacks, at least part of the time, with the guanine ring on the nucleotide residue, G(6), situated in the complementary strand. The binding of Lys-Trp-Lys to (d-CpS(pCpG)2)2 as well as to (d-CpGpCpG)1 was investigated.

  17. Directed site exploration for permeable reactive barrier design

    USGS Publications Warehouse

    Lee, J.; Graettinger, A.J.; Moylan, J.; Reeves, H.W.

    2009-01-01

    Permeable reactive barriers (PRBs) are being employed for in situ site remediation of groundwater that is typically flowing under natural gradients. Site characterization is of critical importance to the success of a PRB. A design-specific site exploration approach called quantitatively directed exploration (QDE) is presented. The QDE approach employs three spatially related matrices: (1) covariance of input parameters, (2) sensitivity of model outputs, and (3) covariance of model outputs to identify the most important location to explore based on a specific design. Sampling at the location that most reduces overall site uncertainty produces a higher probability of success of a particular design. The QDE approach is demonstrated on the Kansas City Plant, Kansas City, MO, a case study where a PRB was installed and failed. It is shown that additional quantitatively directed site exploration during the design phase could have prevented the remedial failure that was caused by missing a geologic body having high hydraulic conductivity at the south end of the barrier. The most contributing input parameter approach using head uncertainty clearly indicated where the next sampling should be made toward the high hydraulic conductivity zone. This case study demonstrates the need to include the specific design as well as site characterization uncertainty when choosing the sampling locations. ?? 2008 Elsevier B.V.

  18. A design method for an intuitive web site

    SciTech Connect

    Quinniey, M.L.; Diegert, K.V.; Baca, B.G.; Forsythe, J.C.; Grose, E.

    1999-11-03

    The paper describes a methodology for designing a web site for human factor engineers that is applicable for designing a web site for a group of people. Many web pages on the World Wide Web are not organized in a format that allows a user to efficiently find information. Often the information and hypertext links on web pages are not organized into intuitive groups. Intuition implies that a person is able to use their knowledge of a paradigm to solve a problem. Intuitive groups are categories that allow web page users to find information by using their intuition or mental models of categories. In order to improve the human factors engineers efficiency for finding information on the World Wide Web, research was performed to develop a web site that serves as a tool for finding information effectively. The paper describes a methodology for designing a web site for a group of people who perform similar task in an organization.

  19. Umatilla Satellite and Release Sites Project : Final Conceptual Design Report.

    SciTech Connect

    Montgomery, James M.

    1992-03-01

    This report presents the results of site analysis for the Umatilla Satellite and Release Sites Project. The purpose of this project is to provide engineering services for the siting and conceptual design of satellite and release facilities for the Umatilla Basin hatchery program. The Umatilla Basin hatchery program consists of artificial production facilities for salmon and steelhead to enhance production in the Umatilla River as defined in the Umatilla master plan approved in 1989 by the Northwest Power Planning Council. Facilities identified in the master plan include adult salmon broodstock holding and spawning facilities, facilities for recovery, acclimation, and/or extended rearing of salmon juveniles, and development of river sites for release of hatchery salmon and steelhead. The historic and current distribution of fall chinook, summer chinook, and coho salmon and steelhead trout was summarized for the Umatilla River basin. Current and future production and release objectives were reviewed. Twenty seven sites were evaluated for the potential development of facilities. Engineering and environmental attributes of the sites were evaluated and compared to facility requirements for water and space. Site screening was conducted to identify the sites with the most potential for facility development. Alternative sites were selected for conceptual design of each facility type. A proposed program for adult holding facilities, final rearing/acclimation, and direct release facilities was developed.

  20. TWRS privatization Phase I site development design requirements document

    SciTech Connect

    Shord, A.L.

    1997-01-21

    The DOE-RL is pursuing a strategy of hiring private contractors for treatment of Hanford Site tank wastes. This strategy is called privatization and includes design, permitting,construction, operation, and deactivation of facilities for tank waste treatment. The TWRS Privatization Infrastructure Project consists of several sub- projects which will provide key services needed to support the privatization mission. This document identifies the design requirements for the site development sub-project, including construction, power, water, and road modifications. It will be used in development of the project`s conceptual design.

  1. Conserved structural chemistry for incision activity in structurally non-homologous apurinic/apyrimidinic endonuclease APE1 and endonuclease IV DNA repair enzymes.

    SciTech Connect

    Tsutakawa, Susan E.; Shin, David S.; Mol, Clifford D.; Izum, Tadahide; Arvai, Andrew S.; Mantha, Anil K.; Szczesny, Bartosz; Ivanov, Ivaylo N.; Hosfield, David J.; Maiti, Buddhadev; Pique, Mike E.; Frankel, Kenneth A.; Hitomi, Kenichi; Cunningham, Richard P.; Mitra, Sankar; Tainer, John A.

    2013-03-22

    Non-coding apurinic/apyrimidinic (AP) sites in DNA form spontaneously and as DNA base excision repair intermediates are the most common toxic and mutagenic in vivo DNA lesion. For repair, AP sites must be processed by 5' AP endonucleases in initial stages of base repair. Human APE1 and bacterial Nfo represent the two conserved 5' AP endonuclease families in the biosphere; they both recognize AP sites and incise the phosphodiester backbone 5' to the lesion, yet they lack similar structures and metal ion requirements. Here, we determined and analyzed crystal structures of a 2.4 ? resolution APE1-DNA product complex with Mg(2+) and a 0.92 Nfo with three metal ions. Structural and biochemical comparisons of these two evolutionarily distinct enzymes characterize key APE1 catalytic residues that are potentially functionally similar to Nfo active site components, as further tested and supported by computational analyses. We observe a magnesium-water cluster in the APE1 active site, with only Glu-96 forming the direct protein coordination to the Mg(2+). Despite differences in structure and metal requirements of APE1 and Nfo, comparison of their active site structures surprisingly reveals strong geometric conservation of the catalytic reaction, with APE1 catalytic side chains positioned analogously to Nfo metal positions, suggesting surprising functional equivalence between Nfo metal ions and APE1 residues. The finding that APE1 residues are positioned to substitute for Nfo metal ions is supported by the impact of mutations on activity. Collectively, the results illuminate the activities of residues, metal ions, and active site features for abasic site endonucleases.

  2. The Effects of Modeled Microgravity on Nucleocytoplasmic Localization of Human Apurinic/Apyrimidinic

    NASA Technical Reports Server (NTRS)

    Gonda, Steve; Jackson, E.B.

    2004-01-01

    Exposure to space radiation and microgravity occurs to humans during space flight. In order to have accurate risk estimations, answering questions to whether increased DNA damage seen during space flight in modified by microgravity are important. Several studies have examined whether intercellular repair of radiation-induced DNA lesions are modified by microgravity. Results from these studies show no modification of the repair processes due to microgravity. However, it is known that in studies not involving radiation that microgravity interferes with normal development. Interestingly, there is no data that attempts to analyze the possible effects of microgravity on the trafficking of DNA repair proteins. In this study, we analyze the effects of modeled microgravity on nucleocytoplasmic shuttling of the human DNA repair enzyme apurinic/apyrimidinic endonuclease 1 (APE1/Ref1) which is involved in base excision repair. We examined nuclear translocation of APE1 using enhanced green fluorescent protein (EGFP) fused to APE1 as a reporter. While APE1 under normal gravity showed normal nuclear localization, APE1 nuclear localization under modeled microgravity was decreased. These results suggest that nucleocytoplasmic translocation of APE1 is modified under modeled microgravity.

  3. Diverse small molecule inhibitors of human apurinic/apyrimidinic endonuclease APE1 identified from a screen of a large public collection.

    PubMed

    Dorjsuren, Dorjbal; Kim, Daemyung; Vyjayanti, Vaddadi N; Maloney, David J; Jadhav, Ajit; Wilson, David M; Simeonov, Anton

    2012-01-01

    The major human apurinic/apyrimidinic endonuclease APE1 plays a pivotal role in the repair of base damage via participation in the DNA base excision repair (BER) pathway. Increased activity of APE1, often observed in tumor cells, is thought to contribute to resistance to various anticancer drugs, whereas down-regulation of APE1 sensitizes cells to DNA damaging agents. Thus, inhibiting APE1 repair endonuclease function in cancer cells is considered a promising strategy to overcome therapeutic agent resistance. Despite ongoing efforts, inhibitors of APE1 with adequate drug-like properties have yet to be discovered. Using a kinetic fluorescence assay, we conducted a fully-automated high-throughput screen (HTS) of the NIH Molecular Libraries Small Molecule Repository (MLSMR), as well as additional public collections, with each compound tested as a 7-concentration series in a 4 µL reaction volume. Actives identified from the screen were subjected to a panel of confirmatory and counterscreen tests. Several active molecules were identified that inhibited APE1 in two independent assay formats and exhibited potentiation of the genotoxic effect of methyl methanesulfonate with a concomitant increase in AP sites, a hallmark of intracellular APE1 inhibition; a number of these chemotypes could be good starting points for further medicinal chemistry optimization. To our knowledge, this represents the largest-scale HTS to identify inhibitors of APE1, and provides a key first step in the development of novel agents targeting BER for cancer treatment.

  4. Diverse Small Molecule Inhibitors of Human Apurinic/Apyrimidinic Endonuclease APE1 Identified from a Screen of a Large Public Collection

    PubMed Central

    Dorjsuren, Dorjbal; Kim, Daemyung; Vyjayanti, Vaddadi N.; Maloney, David J.; Jadhav, Ajit; Wilson, David M.; Simeonov, Anton

    2012-01-01

    The major human apurinic/apyrimidinic endonuclease APE1 plays a pivotal role in the repair of base damage via participation in the DNA base excision repair (BER) pathway. Increased activity of APE1, often observed in tumor cells, is thought to contribute to resistance to various anticancer drugs, whereas down-regulation of APE1 sensitizes cells to DNA damaging agents. Thus, inhibiting APE1 repair endonuclease function in cancer cells is considered a promising strategy to overcome therapeutic agent resistance. Despite ongoing efforts, inhibitors of APE1 with adequate drug-like properties have yet to be discovered. Using a kinetic fluorescence assay, we conducted a fully-automated high-throughput screen (HTS) of the NIH Molecular Libraries Small Molecule Repository (MLSMR), as well as additional public collections, with each compound tested as a 7-concentration series in a 4 µL reaction volume. Actives identified from the screen were subjected to a panel of confirmatory and counterscreen tests. Several active molecules were identified that inhibited APE1 in two independent assay formats and exhibited potentiation of the genotoxic effect of methyl methanesulfonate with a concomitant increase in AP sites, a hallmark of intracellular APE1 inhibition; a number of these chemotypes could be good starting points for further medicinal chemistry optimization. To our knowledge, this represents the largest-scale HTS to identify inhibitors of APE1, and provides a key first step in the development of novel agents targeting BER for cancer treatment. PMID:23110144

  5. Selective inhibition by methoxyamine of the apurinic/apyrimidinic endonuclease activity associated with pyrimidine dimer-DNA glycosylases from Micrococcus luteus and bacteriophage T4

    SciTech Connect

    Liuzzi, M.; Weinfeld, M.; Paterson, M.C.

    1987-06-16

    The UV endonucleases from Micrococcus luteus and bacteriophage T4 possess two catalytic activities specific for the site of cyclobutane pyrimidine dimers in UV-irradiated DNA: a DNA glycosylase that cleaves the 5'-glycosyl bond of the dimerized pyrimidines and an apurinic/apyrimidinic (AP) endonuclease that thereupon incises the phosphodiester bond 3' to the resulting apyrimidinic site. The authors have explored the potential use of methoxyamine, a chemical that reacts at neutral pH with AP sites in DNA, as a selective inhibitor of the AP endonuclease activities residing in the M. luteus and T4 enzymes. The presence of 50 mM methoxyamine during incubation of UV-treated, (/sup 3/H)thymine-labeled poly(dA) x poly(dT) with either enzyme preparation was found to protect completely the irradiated copolymer from endonucleolytic attack at dimer sites, as assayed by yield of acid-soluble radioactivity. In contrast, the dimer-DNA glycosylase activity of each enzyme remained fully functional, as monitored retrospectively by release of free thymine after either photochemical-(5 kJ/m/sup 2/, 254 nm) or photoenzymic- (Escherichia coli photolyase plus visible light) induced reversal of pyrimidine dimers in the UV-damaged substrate. The data demonstrate that the inhibition of the strand-incision reaction arises because of chemical modification of the AP sites and is not due to inactivation of the enzyme by methoxyamine. The results, combined with earlier findings for 5'-acting AP endonucleases, strongly suggest that methoxyamine is a highly specific inhibitor of virtually all AP endonucleases, irrespective of their modes of action, and may therefore prove useful in a wide variety of DNA repair studies.

  6. Structure-based virtual screening toward the discovery of novel inhibitors of the DNA repair activity of the human apurinic/apyrimidinic endonuclease 1.

    PubMed

    Guerreiro, Patrícia S; Estácio, Sílvia G; Antunes, Fernando; Fernandes, Ana S; Pinheiro, Pedro F; Costa, João G; Castro, Matilde; Miranda, Joana P; Guedes, Rita C; Oliveira, Nuno G

    2016-12-01

    The DNA repair activity of human apurinic/apyrimidinic endonuclease 1 (APE1) has been recognized as a promising target for the development of small-molecule inhibitors to be used in combination with anticancer agents. In an attempt to identify novel inhibitors of APE1, we present a structure-based virtual screening (SBVS) study based on molecular docking analysis of the compounds of NCI database using the GOLD 5.1.0 (Genetic Optimization for Ligand Docking) suite of programs. Compounds selected in this screening were tested with a fluorescence-based APE1 endonuclease activity assay. Two compounds (37 and 41) were able to inhibit the multifunctional enzyme APE1 in the micromolar range, while compound 22 showed inhibitory effects at nanomolar concentrations. These results were confirmed by a plasmid DNA nicking assay. In addition, the potential APE1 inhibitors did not affect the cell viability of non-tumor MCF10A cells. Overall, compounds 22, 37, and 41 appear to be important scaffolds for the design of novel APE1 inhibitors and this study highlights the relevance of in silico-based approaches as valuable tools in drug discovery. © 2016 John Wiley & Sons A/S.

  7. Remedial design of the Fultz Landfill Site, Byesville, Ohio

    SciTech Connect

    Rajaram, V.; Riesing, R.; Bloom, T.

    1994-12-31

    The Fultz Landfill Superfund (Fultz) site is a 30-acre hazardous waste landfill located near Byesville, Ohio. The site is approximately 75 miles east of Columbus and 3 miles southwest of Cambridge, the largest city in Guernsey County, Ohio. The landfill is situated on the north slope of a ridge that overlies abandoned coal mines in the Upper Freeport Coal seam. The north half of the landfill lies in an unreclaimed strip mine in the Upper Freeport Coal seam, where saturated portions of surface mine spoils and natural soils form the ``shallow aquifer``. The south half of the landfill lies 40 to 50 feet (ft.) above an abandoned, flooded deep mine in the same coal seam. The flooded deep mine forms an aquifer referred to as the ``coal mine aquifer``. This paper presents the results of design studies completed by PRC Environmental Management, Inc. (PRC), during 1993, and the remedial design (RD) of the components specified by the US Environmental Protection Agency (EPA) Record of Decision (ROD) for the Fultz site (EPA 1991). The remedy specified in the ROD includes a multilayer landfill cap that is compliant with Resource Conservation and Recovery Act (RCRA) Subtitle C guidelines, a leachate collection and groundwater extraction and treatment system, and stabilizing mine voids underlying the southern portion of the site. Vinyl chloride is the only contaminant exceeding a maximum contaminant limit (MCL) in the coal mine aquifer.

  8. The Hanford Site 1000-Year Cap Design Test

    SciTech Connect

    Gee, Glendon W. ); Ward, Anderson L. ); Wittreich, Curtis D.

    2002-12-27

    Surface barrier or capping technology is needed to isolate buried wastes. A successful cap must prevent the intrusion of plants, animals, and man into the underlying waste, minimize wind and water erosion, require minimal maintenance, and limit water intrusion to near-zero amounts. For some sites where wastes are long-lived, caps should potentially last a thousand years or more. At the U.S. Department of Energy (DOE) Hanford Site in Washington State, a surface cap with a 1000-year design life was constructed and then tested and monitored for performance under wetting conditions that are extreme for the region. The cap was built in 1994 over an existing waste site as a part of a Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) treatability test. The above-grade barrier or cap consists of a 2-m-thick silt-loam soil overlying layers (from top down) of sand, gravel, basalt rock (riprap), and a low-permeability asphalt. Two sideslope configurations, a clean-fill gravel on a 10:1 slope and a basalt riprap on a 2:1 slope were part of the overall design and testing. Design considerations included constructability; water-balance monitoring; wind and water erosion control and monitoring; surface revegetation, biointrusion control, subsidence, and sideslope stability; and durability of the asphalt layer.

  9. Optimising web site designs for people with learning disabilities

    PubMed Central

    Williams, Peter; Hennig, Christian

    2015-01-01

    Much relevant internet-mediated information is inaccessible to people with learning disabilities because of difficulties in navigating the web. This paper reports on the methods undertaken to determine how information can be optimally presented for this cohort. Qualitative work is outlined where attributes relating to site layout affecting usability were elicited. A study comparing web sites of different design layouts exhibiting these attributes is discussed, with the emphasis on methodology. Eight interfaces were compared using various combinations of menu position (vertical or horizontal), text size and the absence or presence of images to determine which attributes of a site have the greatest performance impact. Study participants were also asked for their preferences, via a ‘smiley-face’ rating scale and simple interviews. ‘Acquiescence bias’ was minimised by avoiding polar (‘yes/no’) interrogatives, achieved by asking participants to compare layouts (such as horizontal versus vertical menu), with reasons coaxed from those able to articulate them. Preferred designs were for large text and images. This was the reverse of those facilitating fastest retrieval times, a discrepancy due to preferences being judged on aesthetic considerations. Design recommendations that reconcile preference and performance findings are offered. These include using a horizontal menu, juxtaposing images and text, and reducing text from sentences to phrases, thus facilitating preferred large text without increasing task times. PMID:26097431

  10. Optimising web site designs for people with learning disabilities.

    PubMed

    Williams, Peter; Hennig, Christian

    2015-01-01

    Much relevant internet-mediated information is inaccessible to people with learning disabilities because of difficulties in navigating the web. This paper reports on the methods undertaken to determine how information can be optimally presented for this cohort. Qualitative work is outlined where attributes relating to site layout affecting usability were elicited. A study comparing web sites of different design layouts exhibiting these attributes is discussed, with the emphasis on methodology. Eight interfaces were compared using various combinations of menu position (vertical or horizontal), text size and the absence or presence of images to determine which attributes of a site have the greatest performance impact. Study participants were also asked for their preferences, via a 'smiley-face' rating scale and simple interviews. 'Acquiescence bias' was minimised by avoiding polar ('yes/no') interrogatives, achieved by asking participants to compare layouts (such as horizontal versus vertical menu), with reasons coaxed from those able to articulate them. Preferred designs were for large text and images. This was the reverse of those facilitating fastest retrieval times, a discrepancy due to preferences being judged on aesthetic considerations. Design recommendations that reconcile preference and performance findings are offered. These include using a horizontal menu, juxtaposing images and text, and reducing text from sentences to phrases, thus facilitating preferred large text without increasing task times.

  11. Coordination of Steps in Single-nucleotide Base Excision Repair Mediated by Apurinic/Apyrimidinic Endonuclease 1 and DNA Polymerase β*

    PubMed Central

    Liu, Yuan; Prasad, Rajendra; Beard, William A.; Kedar, Padmini S.; Hou, Esther W.; Shock, David D.; Wilson, Samuel H.

    2008-01-01

    The individual steps in single-nucleotide base excision repair (SN-BER) are coordinated to enable efficient repair without accumulation of cytotoxic DNA intermediates. The DNA transactions and various proteins involved in SN-BER of abasic sites are well known in mammalian systems. Yet, despite a wealth of information on SN-BER, the mechanism of step-by-step coordination is poorly understood. In this study we conducted experiments toward understanding step-by-step coordination during BER by comparing DNA binding specificities of two major human SN-BER enzymes, apurinic/aprymidinic endonuclease 1 (APE) and DNA polymerase β (Pol β). It is known that these enzymes do not form a stable complex in solution. For each enzyme, we found that DNA binding specificity appeared sufficient to explain the sequential processing of BER intermediates. In addition, however, we identified at higher enzyme concentrations a ternary complex of APE·Pol β·DNA that formed specifically at BER intermediates containing a 5′-deoxyribose phosphate group. Formation of this ternary complex was associated with slightly stronger Pol β gap-filling and much stronger 5′-deoxyribose phosphate lyase activities than was observed with the Pol β·DNA binary complex. These results indicate that step-by-step coordination in SN-BER can rely on DNA binding specificity inherent in APE and Pol β, although coordination also may be facilitated by APE·Pol β·DNA ternary complex formation with appropriate enzyme expression levels or enzyme recruitment to sites of repair. PMID:17355977

  12. Insight into mechanisms of 3′-5′ exonuclease activity and removal of bulky 8,5′-cyclopurine adducts by apurinic/apyrimidinic endonucleases

    PubMed Central

    Mazouzi, Abdelghani; Vigouroux, Armelle; Aikeshev, Bulat; Brooks, Philip J.; Saparbaev, Murat K.; Morera, Solange; Ishchenko, Alexander A.

    2013-01-01

    8,5′-cyclo-2'-deoxyadenosine (cdA) and 8,5′-cyclo-2'-deoxyguanosine generated in DNA by both endogenous oxidative stress and ionizing radiation are helix-distorting lesions and strong blocks for DNA replication and transcription. In duplex DNA, these lesions are repaired in the nucleotide excision repair (NER) pathway. However, lesions at DNA strand breaks are most likely poor substrates for NER. Here we report that the apurinic/apyrimidinic (AP) endonucleases—Escherichia coli Xth and human APE1—can remove 5′S cdA (S-cdA) at 3′ termini of duplex DNA. In contrast, E. coli Nfo and yeast Apn1 are unable to carry out this reaction. None of these enzymes can remove S-cdA adduct located at 1 or more nt away from the 3′ end. To understand the structural basis of 3′ repair activity, we determined a high-resolution crystal structure of E. coli Nfo-H69A mutant bound to a duplex DNA containing an α-anomeric 2′-deoxyadenosine:T base pair. Surprisingly, the structure reveals a bound nucleotide incision repair (NIR) product with an abortive 3′-terminal dC close to the scissile position in the enzyme active site, providing insight into the mechanism for Nfo-catalyzed 3′→5′ exonuclease function and its inhibition by 3′-terminal S-cdA residue. This structure was used as a template to model 3′-terminal residues in the APE1 active site and to explain biochemical data on APE1-catalyzed 3′ repair activities. We propose that Xth and APE1 may act as a complementary repair pathway to NER to remove S-cdA adducts from 3′ DNA termini in E. coli and human cells, respectively. PMID:23898172

  13. DNA base-excision repair enzyme apurinic/apyrimidinic endonuclease/redox factor-1 is increased and competent in the brain and spinal cord of individuals with amyotrophic lateral sclerosis.

    PubMed

    Shaikh, Arif Y; Martin, Lee J

    2002-01-01

    Motor neurons degenerate in amyotrophic lateral sclerosis (ALS). The mechanisms for this neuronal cell death are not known, although apoptosis has been implicated. Oxidative damage to DNA and activation of p53 has been identified directly in motor neurons in cases of ALS. We evaluated whether motor neuron degeneration in ALS is associated with changes in the levels and function of the multifunctional protein apurinic/apyrimidinic endonuclease (APE/Ref-1). APE/Ref-1 functions as an enzyme in the DNA base-excision repair pathway and as a redox-regulation protein for transcription factors. The protein level and localization of APE/Ref-1 are changed in ALS. Immunoblotting showed that APE/Ref-1 protein levels are increased in selectively vulnerable central nervous system (CNS) regions in individuals with ALS compared to age-matched controls. Plasmid DNA repair assay demonstrated that APE from individuals with ALS is competent in repairing apurinic (AP) sites. DNA repair function in nuclear fractions is increased significantly in ALS motor cortex and spinal cord. Immunocytochemistry and single-cell densitometry revealed that APE/Ref-1 is expressed at lower levels in control motor neurons than in ALS motor neurons, which are decreased in number by 42% in motor cortex. APE/Ref-1 is increased in the nucleus of remaining upper motor neurons in ALS, which show a 38% loss of nuclear area. APE-Ref-1 is also upregulated in astrocytes in spinal cord white matter pathways in familial ALS. We conclude that mechanisms for DNA repair are activated in ALS, supporting the possibility that DNA damage is an upstream mechanism for motor neuron degeneration in this disease.

  14. Remedial action plan and site design for stabilization of the inactive uranium mill tailings sites at Rifle, Colorado

    SciTech Connect

    Not Available

    1992-02-01

    This appendix assesses the present conditions and data gathered about the two inactive uranium mill tailings sites near Rifle, Colorado, and the designated disposal site six miles north of Rifle in the area of Estes Gulch. It consolidates available engineering, radiological, geotechnical, hydrological, meteorological, and other information pertinent to the design of the Remedial Action Plan (RAP). The data characterize conditions at the mill, tailings, and disposal site so that the Remedial Action Contractor (RAC) may complete final designs for the remedial actions.

  15. Brownfields Green Avenue Sites: Technical Memorandum - Conceptual Design for Sustainable Redevelopment

    EPA Pesticide Factsheets

    This technical memorandum briefly describes the site and proposed conceptual site plan, indicates conceptual design considerations, specifies recommended green and sustainable features, and offers other recommendations

  16. Probabilistic envelope curves for design flood estimation at ungauged sites

    NASA Astrophysics Data System (ADS)

    Castellarin, Attilio

    2007-04-01

    Castellarin et al. (2005) introduced the probabilistic regional envelope curves (PRECs) of flood flows and formulated an empirical estimator of the recurrence interval T associated with the PREC for M synthetic cross-correlated and concurrent sequences of annual maxima with length n. In principle, a PREC can be used to estimate the T-year flood (design flood) for any basin in a given region as a function of the catchment area alone. A number of fundamental issues hamper the construction of PRECs and the estimation of their T for historical regional samples of flood flows. Some of these issues are addressed by this study, which presents an algorithm for the application of the empirical estimator of T to historical annual maximum series of unequal length and assesses how the selection of a particular cross-correlation formula and plotting position affects the accuracy of design flood estimates retrieved from PRECs (PREC flood quantiles). The reliability of the proposed algorithm and the accuracy of the flood quantiles are discussed through a comprehensive cross validation. The results show that the reliability of PREC flood quantiles for ungauged sites is comparable with the reliability of regional estimates produced by the application of the index flood approach. The results of the cross-validation also show that the selection of a particular correlation formula or plotting position may produce biased PREC flood quantiles, and suggest that the accuracy of the approach could be significantly enhanced by relaxing the assumption that the design flood scales with catchment area alone.

  17. Fly ash design manual for road and site applications

    SciTech Connect

    DiGioia, A.M. Jr.; Brendel, G.F.; McLaren, R.J.; Balsamo, N.J.; Glogowski, P.E.; Kelley, J.M. )

    1992-04-01

    This design manual is the second of two volumes pertaining to the use of fly ash as a construction material in road and site development applications. Volume 1 describes the use of fly ash conditioned with small amounts of water to produce a solid, soil-like material. Volume 2 describes the use of fly ash mixed with sufficient water to create a fluid or semi-fluid material. In most applications, Portland cement is added to the mixture to provide the desired level of strength. In some applications, aggregate filler is also added to the mixture to improve the durability of the mix. The flowable nature of these materials determines the manner in which they are handled, placed, and used in construction. The following uses are discussed: Slurry Backfills; Grouting; and Hydraulic Fills. Miscellaneous uses are also discussed, including the use of fly ash in high fly ash content concrete, unsintered synthetic aggregate, and slurry walls. Also included are three appendices containing additional information of interest to the designer: Mix Design; Laboratory Test Procedures; and a Bibliography.

  18. Designing for Differences: Cultural Issues in the Design of WWW-based Course-support Sites.

    ERIC Educational Resources Information Center

    Collis, Betty

    1999-01-01

    Examines factors affecting the cultural appropriateness of Web-based course-support sites. Identifies a strategy for accommodating different values of these factors in Web-based course-support systems, based on a set of 10 design guidelines. Describes and evaluates the TeleTOP Method from the University of Twente (The Netherlands) to illustrate…

  19. A reagentless electrochemiluminescent immunosensor for apurinic/apyrimidinic endonuclease 1 detection based on the new Ru(bpy)3(2+)/bi-arginine system.

    PubMed

    Zhao, Min; Chai, Xi Deng Ya-Qin; Han, Jing; Gui, Guo-Feng; Yuan, Ruo; Zhuo, Ying

    2014-10-10

    Apurinic/apyrimidinic endonuclease 1 (APE-1), a kind of multifunctional protein widely-distributed in the body, plays an essential role in the DNA base excision repair and serves as multiple possible roles in the response of human cancer to radiotherapy and chemotherapy. In this work, an ultrasensitive solid-state electrochemiluminescence (ECL) immunosensor is designed to determine APE-1 based on the new Ru(bpy)3(2+)/bi-arginine system. The bi-arginine (bi-Arg) is decorated on the Au nanoparticles functionalized magnetic Fe3O4/reduced graphene oxide (bi-Arg/Au@Fe3O4-rGO) according to the self-assembling and covalent cross-linking interaction to obtain the functionalized nanocomposite of bi-Arg/Au@Fe3O4-rGO. Herein, the bi-Arg/Au@Fe3O4-rGO plays not only an amplification label to enhance the ECL signal of Ru(bpy)3(2+) due to the coreactant of bi-Arg but also an ideal nanocarrier to load numerous secondary antibody. Based on sandwich-type immunoassay format, this proposed method offers a linear range of 1.0fgmL(-1)-5.0pgmL(-1) and an estimated detection limit of 0.3fgmL(-1) for the APE-1. Moreover, the reagentless ECL immunosensor also exhibits high sensitivity, excellent selectivity and good stability, which has greatly potential development and application in clinical diagnostics, immunology and biomedical research.

  20. Remedial action plan and site design for stabilization of the inactive uranium mill tailings site at Tuba City, Arizona. Apendix D, Site characteriztion

    SciTech Connect

    Not Available

    1987-05-01

    This appendix assesses the present conditions and data for the inactive uranium mill site near Tuba City, Arizona. It consolidates available engineering, radiological, geotechnical, hydrological, meterological, and other information pertinent to the design of the Remedial Action Plan (RAP). The data characterize conditions at the mill and tailings site so that the Remedial Action Contractor (RAC) may complete final designs of the remedial actions.

  1. 78 FR 37759 - Ocean Dumping; Atchafalaya-West Ocean Dredged Material Disposal Site Designation

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-06-24

    ... AGENCY 40 CFR Part 228 Ocean Dumping; Atchafalaya-West Ocean Dredged Material Disposal Site Designation... designate the Atchafalaya-West Ocean Dredged Material Disposal Site pursuant to the draft EIS, ``Designation of the Atchafalaya River Bar Channel Ocean Dredged Material Disposal Site Pursuant to Section 102(c...

  2. 78 FR 29687 - Ocean Dumping; Atchafalaya-West Ocean Dredged Material Disposal Site Designation

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-05-21

    ... AGENCY 40 CFR Part 228 Ocean Dumping; Atchafalaya-West Ocean Dredged Material Disposal Site Designation...) Atchafalaya-West Ocean Disposal Site (ODMDS-West) as a permanent MPRSA Section 102(c) ocean dredged material... Management and Monitoring Plan E. Ocean Dumping Site Designation Criteria --General Selection Criteria...

  3. A Pilot Sampling Design for Estimating Outdoor Recreation Site Visits on the National Forests

    Treesearch

    Stanley J. Zarnoch; S.M. Kocis; H. Ken Cordell; D.B.K. English

    2002-01-01

    A pilot sampling design is described for estimating site visits to National Forest System lands. The three-stage sampling design consisted of national forest ranger districts, site days within ranger districts, and last-exiting recreation visitors within site days. Stratification was used at both the primary and secondary stages. Ranger districts were stratified based...

  4. 40 CFR 228.15 - Dumping sites designated on a final basis.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 24 2010-07-01 2010-07-01 false Dumping sites designated on a final basis. 228.15 Section 228.15 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) OCEAN DUMPING CRITERIA FOR THE MANAGEMENT OF DISPOSAL SITES FOR OCEAN DUMPING § 228.15 Dumping sites designated on a final basis. (a)(1) The...

  5. Designing a Marketing Course with Field Site Visits

    ERIC Educational Resources Information Center

    Van Doren, Doris; Corrigan, Hope Bober

    2008-01-01

    A key goal of including field site visits in marketing courses is to give business students increased interaction with industry professionals and community leaders. Site visits give students a concrete idea of how different marketing disciplines work in the business world. Business students gain greater insight into a career in marketing from this…

  6. 40 CFR 228.4 - Procedures for designation of sites.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... waste disposal on areas similar to such sites in physical, chemical, and biological characteristics. All... to the specific conditions of Dredged Material permits issued by the U.S. Army Corps of Engineers... dredged material disposal on areas similar to such sites in physical, chemical, and biological...

  7. 40 CFR 228.4 - Procedures for designation of sites.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... waste disposal on areas similar to such sites in physical, chemical, and biological characteristics. All... to the specific conditions of Dredged Material permits issued by the U.S. Army Corps of Engineers... dredged material disposal on areas similar to such sites in physical, chemical, and biological...

  8. Designing a Marketing Course with Field Site Visits

    ERIC Educational Resources Information Center

    Van Doren, Doris; Corrigan, Hope Bober

    2008-01-01

    A key goal of including field site visits in marketing courses is to give business students increased interaction with industry professionals and community leaders. Site visits give students a concrete idea of how different marketing disciplines work in the business world. Business students gain greater insight into a career in marketing from this…

  9. Fractionated Radiation Exposure of Rat Spinal Cords Leads to Latent Neuro-Inflammation in Brain, Cognitive Deficits, and Alterations in Apurinic Endonuclease 1.

    PubMed

    Suresh Kumar, M A; Peluso, Michael; Chaudhary, Pankaj; Dhawan, Jasbeer; Beheshti, Afshin; Manickam, Krishnan; Thapar, Upasna; Pena, Louis; Natarajan, Mohan; Hlatky, Lynn; Demple, Bruce; Naidu, Mamta

    2015-01-01

    Ionizing radiation causes degeneration of myelin, the insulating sheaths of neuronal axons, leading to neurological impairment. As radiation research on the central nervous system has predominantly focused on neurons, with few studies addressing the role of glial cells, we have focused our present research on identifying the latent effects of single/ fractionated -low dose of low/ high energy radiation on the role of base excision repair protein Apurinic Endonuclease-1, in the rat spinal cords oligodendrocyte progenitor cells' differentiation. Apurinic endonuclease-1 is predominantly upregulated in response to oxidative stress by low- energy radiation, and previous studies show significant induction of Apurinic Endonuclease-1 in neurons and astrocytes. Our studies show for the first time, that fractionation of protons cause latent damage to spinal cord architecture while fractionation of HZE (28Si) induce increase in APE1 with single dose, which then decreased with fractionation. The oligodendrocyte progenitor cells differentiation was skewed with increase in immature oligodendrocytes and astrocytes, which likely cause the observed decrease in white matter, increased neuro-inflammation, together leading to the observed significant cognitive defects.

  10. Fractionated Radiation Exposure of Rat Spinal Cords Leads to Latent Neuro-Inflammation in Brain, Cognitive Deficits, and Alterations in Apurinic Endonuclease 1

    DOE PAGES

    Suresh Kumar, M. A.; Peluso, Michael; Chaudhary, Pankaj; ...

    2015-07-24

    Ionizing radiation causes degeneration of myelin, the insulating sheaths of neuronal axons, leading to neurological impairment. As radiation research on the central nervous system has predominantly focused on neurons, with few studies addressing the role of glial cells, we have focused our present research on identifying the latent effects of single/ fractionated -low dose of low/ high energy radiation on the role of base excision repair protein Apurinic Endonuclease-1, in the rat spinal cords oligodendrocyte progenitor cells ’ differentiation. Apurinic endonuclease-1 is predominantly upregulated in response to oxidative stress by low- energy radiation, and previous studies show significant induction ofmore » Apurinic Endonucle- ase-1 in neurons and astrocytes. Our studies show for the first time, that fractionation of pro- tons cause latent damage to spinal cord architecture while fractionation of HZE (28Si) induce increase in APE1 with single dose, which then decreased with fractionation. In conclusion, the oligoden- drocyte progenitor cells differentiation was skewed with increase in immature oligodendro- cytes and astrocytes, which likely cause the observed decrease in white matter, increased neuro-inflammation, together leading to the observed significant cognitive defects« less

  11. Fractionated Radiation Exposure of Rat Spinal Cords Leads to Latent Neuro-Inflammation in Brain, Cognitive Deficits, and Alterations in Apurinic Endonuclease 1

    SciTech Connect

    Suresh Kumar, M. A.; Peluso, Michael; Chaudhary, Pankaj; Dhawan, Jasbeer; Beheshti, Afshin; Manickam, Krishnan; Thapar, Upasna; Pena, Louis; Natarajan, Mohan; Hlatky, Lynn; Demple, Bruce; Naidu, Mamta

    2015-07-24

    Ionizing radiation causes degeneration of myelin, the insulating sheaths of neuronal axons, leading to neurological impairment. As radiation research on the central nervous system has predominantly focused on neurons, with few studies addressing the role of glial cells, we have focused our present research on identifying the latent effects of single/ fractionated -low dose of low/ high energy radiation on the role of base excision repair protein Apurinic Endonuclease-1, in the rat spinal cords oligodendrocyte progenitor cells ’ differentiation. Apurinic endonuclease-1 is predominantly upregulated in response to oxidative stress by low- energy radiation, and previous studies show significant induction of Apurinic Endonucle- ase-1 in neurons and astrocytes. Our studies show for the first time, that fractionation of pro- tons cause latent damage to spinal cord architecture while fractionation of HZE (28Si) induce increase in APE1 with single dose, which then decreased with fractionation. In conclusion, the oligoden- drocyte progenitor cells differentiation was skewed with increase in immature oligodendro- cytes and astrocytes, which likely cause the observed decrease in white matter, increased neuro-inflammation, together leading to the observed significant cognitive defects

  12. User-Centered Design and Usability Testing of a Web Site: An Illustrative Case Study.

    ERIC Educational Resources Information Center

    Corry, Michael D.; Frick, Theodore W.; Hansen, Lisa

    1997-01-01

    Presents an overview of user-centered design and usability testing. Describes a Web site evaluation project at a university, the iterative process of rapid prototyping and usability testing, and how the findings helped to improve the design. Discusses recommendations for university Web site design and reflects on problems faced in usability…

  13. User-Centered Design and Usability Testing of a Web Site: An Illustrative Case Study.

    ERIC Educational Resources Information Center

    Corry, Michael D.; Frick, Theodore W.; Hansen, Lisa

    1997-01-01

    Presents an overview of user-centered design and usability testing. Describes a Web site evaluation project at a university, the iterative process of rapid prototyping and usability testing, and how the findings helped to improve the design. Discusses recommendations for university Web site design and reflects on problems faced in usability…

  14. Criteria for the management of disposal sites for ocean dumping extension of interim designations

    SciTech Connect

    Not Available

    1980-01-16

    The US Environmental Protection Agency has promulgated amendments to its regulations on ocean dumping which extend the interim designation of certain ocean dumping sites pending completion of Environmental Impact Statements (EIS) and formal rulemaking procedures. Other sites, mainly those for dredged material disposal, will be extended pending completion of site designation studies and formal designation. The schedule for availability of draft EIS on 25 dumping sites and extension dates for the affected interim sites are discussed. This rule is effective as of 1/16/80. Comments must be received by 2/15/80.

  15. Energy conserving site design case study: Shenandoah, Georgia. Final report

    SciTech Connect

    Not Available

    1980-01-01

    The case study examines the means by which energy conservation can be achieved at an aggregate community level by using proper planning and analytical techniques for a new town, Shenandoah, Georgia, located twenty-five miles southwest of Atlanta's Hartsfield International Airport. A potentially implementable energy conservation community plan is achieved by a study team examining the land use options, siting characteristics of each building type, alternate infrastructure plans, possible decentralized energy options, and central utility schemes to determine how community energy conservation can be achieved by use of pre-construction planning. The concept for the development of mixed land uses as a passively sited, energy conserving community is based on a plan (Level 1 Plan) that uses the natural site characteristics, maximizes on passive energy siting requirement, and allows flexibility for the changing needs of the developers. The Level 2 Plan is identical with Level 1 plan plus a series of decentraized systems that have been added to the residential units: the single-family detached, the apartments, and the townhouses. Level 3 Plan is similar to the Level 1 Plan except that higher density dwellings have been moved to areas adjacent to central site. The total energy savings for each plan relative to the conventional plan are indicated. (MCW)

  16. Optimising Web Site Designs for People with Learning Disabilities

    ERIC Educational Resources Information Center

    Williams, Peter; Hennig, Christian

    2015-01-01

    Much relevant internet-mediated information is inaccessible to people with learning disabilities because of difficulties in navigating the web. This paper reports on the methods undertaken to determine how information can be optimally presented for this cohort. Qualitative work is outlined where attributes relating to site layout affecting…

  17. Restructuring Web Site Design: An Alternative to WebTeams.

    ERIC Educational Resources Information Center

    Davis, Marc; Boyer, Janice

    In July 1997, the University Library at the University of Nebraska at Omaha adopted a new structural model for Web site development--the publishing metaphor. Rather than relying on an individual expert (the Webmaster) or a WebTeam comprised of a few individuals, the model permits everyone in the organization, regardless of position or Web…

  18. Basic repository environmental assessment design basis: Deaf Smith County site

    SciTech Connect

    Not Available

    1988-03-01

    This study examines the engineering factors and costs associated with the construction, operation, and decommissioning of a high-level nuclear waste repository in salt in the Palo Duro Basin in Deaf Smith County, Texas. The study assumes a repository capacity of 36,000 metric tons of heavy metal (MTHM) of unreprocessed spent fuel and 36,000 MTHM of commercial high-level reprocessing waste, along with 7,020 canisters of defense high-level reprocessing waste and associated quantities or remote- and contact-handled transuranic waste (TRU). With the exception of TRU, all the waste forms are placed in 300- to 1000-year-life carbon-steel waste packages in a collocated waste handling and packaging facility (WHPF), which is also described. The construction, operation, and decommissioning of the proposed repository is estimated to cost approximately $4.64 billion. Costs include those for the collocate WHPF, engineering, and contingency, but exclude waste form assembly and shipment to the site and waste package fabrication and shipment to the site. These costs reflect the relative average wage rates of the region, the relatively easy access to the site, and the relatively weak nature of the salt at this site. Construction would require an estimated 7 to 7.5 years. Engineering factors and costs are not strongly influenced by environmental considerations. 62 refs., 24 figs., 20 tabs.

  19. Optimising Web Site Designs for People with Learning Disabilities

    ERIC Educational Resources Information Center

    Williams, Peter; Hennig, Christian

    2015-01-01

    Much relevant internet-mediated information is inaccessible to people with learning disabilities because of difficulties in navigating the web. This paper reports on the methods undertaken to determine how information can be optimally presented for this cohort. Qualitative work is outlined where attributes relating to site layout affecting…

  20. Down-regulation of apurinic/apyrimidinic endonuclease 1 (APE1) in spinal motor neurones under oxidative stress.

    PubMed

    Chu, Tak-Ho; Guo, Anchen; Wu, Wutian

    2014-06-01

    Apurinic/apyrimidinic endonuclease 1 (APE1) is an intermediate enzyme in base excision repair which is important for removing damaged nucleotides under normal and pathological conditions. Accumulation of damaged bases causes genome instability and jeopardizes cell survival. Our study is to examine APE1 regulation under oxidative stress in spinal motor neurones which are vulnerable to oxidative insult. We challenged the motor neurone-like cell line NSC-34 with hydrogen peroxide and delineated APE1 function by applying various inhibitors. We also examined the expression of APE1 in spinal motor neurones after spinal root avulsion in adult rats. We showed that hydrogen peroxide induced APE1 down-regulation and cell death in a differentiated motor neurone-like cell line. Inhibiting the two functional domains of APE1, namely, DNA repair and redox domains potentiated hydrogen peroxide induced cell death. We further showed that p53 phosphorylation early after hydrogen peroxide treatment might contribute to the down-regulation of APE1. Our in vivo results similarly showed that APE1 was down-regulated after root avulsion injury in spinal motor neurones. Delay of motor neurone death suggested that APE1 might not cause immediate cell death but render motor neurones vulnerable to further oxidative insults. We conclude that spinal motor neurones down-regulate APE1 upon oxidative stress. This property renders motor neurones susceptible to continuous challenge of oxidative stress in pathological conditions. © 2013 British Neuropathological Society.

  1. Roadmap for Integrating Sustainable Design into Site-Level Operations

    SciTech Connect

    Peterson, Keith L.; Dorsey, Judy A.

    2000-04-19

    Sustainable Design recognizes that products and processes are interdependent with the environmental, economic, and social systems surrounding them and implements measures to prevent an unsustainable compromise to these systems.

  2. ENGINEERING BULLETIN: DESIGN CONSIDERATIONS FOR AMBIENT AIR MONITORING AT SUPERFUND SITES

    EPA Science Inventory

    The Engineering Bulletin is intended to help the RPM design the site-specific air monitoring program needed before and during site remediation. The types of ambient air monitoring activities of interest at Superfund sites are selecting the most appropriate approach, establishing ...

  3. Site Design Applications for the Theater Construction Management System

    DTIC Science & Technology

    1993-10-01

    wherever possible, thereby allowing the Army to use the il best software available without directly absorbing the costs JAN 0 4 1994 associated with... practices were reviewed; the requirements and conceptual basis for such a software integration were developed; and two potential ad- vanced technology site...allowing the Army to use the best software available without directly absorbing the costs associated with software development. This study

  4. Basic repository environmental assessment design basis, Lavender Canyon site

    SciTech Connect

    Not Available

    1988-01-01

    This study examines the engineering factors and costs associated with the construction, operation, and decommissioning of a high-level nuclear waste repository in salt in the Paradox Basin in Lavender Canyon, Utah. The study assumes a repository capacity of 36,000 metric tons of heavy metal (MTHM) of unreprocessed spent fuel and 36,000 MTHM of commercial high-level reprocessing waste, along with 7020 canisters of defense high-level reprocessing waste and associated quantities of remote- and contact-handled transuranic waste (TRU). With the exception of TRU, all the waste forms are placed in 300- to 1000-year-life carbon-steel waste packages in a collocated waste handling and packaging facility (WHPF), which is also described. The construction, operation, and decommissioning of the proposed repository is estimated to cost approximately $5.51 billion. Costs include those for the collocated WHPP, engineering, and contingency, but exclude waste form assembly and shipment to the site and waste package fabrication and shipment to the site. These costs reflect the relative average wage rates of the region and the relatively sound nature of the salt at this site. Construction would require an estimated 7.75 years. Engineering factors and costs are not strongly influenced by environmental considerations. 51 refs., 24 figs., 20 tabs.

  5. Campus Demonstration Sites for Sustainable Systems and Design: Five "Creation" Stories.

    ERIC Educational Resources Information Center

    Jack, Kathy; Ihara, Dan, Ed.

    This paper provides a summary of the development and management of five campus demonstration sites designed to create harmony with natural systems and meet current student needs without compromising the needs of future generations. Information for each campus includes an overview of the site, project origins, the proposal and design process, the…

  6. New "persona" concept helps site designers cater to target user segments' needs.

    PubMed

    2004-09-01

    Using the relatively new "persona" design concept, Web strategists create a set of archetypical user characters, each one representing one of their site's primary audiences. Then, as their site is constructed or upgraded, they champion the personas, arguing on their behalf and forcing the design team to take each audience's needs and wants into account.

  7. 24 CFR 3285.103 - Site suitability with design zone maps.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... maps. 3285.103 Section 3285.103 Housing and Urban Development Regulations Relating to Housing and Urban... Considerations § 3285.103 Site suitability with design zone maps. Prior to the initial installation of a new... indicated on the design zone maps provided with the home, are suitable for the site location where the...

  8. 10 CFR 61.51 - Disposal site design for land disposal.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 10 Energy 2 2011-01-01 2011-01-01 false Disposal site design for land disposal. 61.51 Section 61.51 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSING REQUIREMENTS FOR LAND DISPOSAL OF RADIOACTIVE WASTE Technical Requirements for Land Disposal Facilities § 61.51 Disposal site design for...

  9. 10 CFR 61.51 - Disposal site design for land disposal.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 10 Energy 2 2010-01-01 2010-01-01 false Disposal site design for land disposal. 61.51 Section 61.51 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSING REQUIREMENTS FOR LAND DISPOSAL OF RADIOACTIVE WASTE Technical Requirements for Land Disposal Facilities § 61.51 Disposal site design for...

  10. 10 CFR 61.51 - Disposal site design for land disposal.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 10 Energy 2 2014-01-01 2014-01-01 false Disposal site design for land disposal. 61.51 Section 61.51 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSING REQUIREMENTS FOR LAND DISPOSAL OF RADIOACTIVE WASTE Technical Requirements for Land Disposal Facilities § 61.51 Disposal site design for...

  11. 10 CFR 61.51 - Disposal site design for land disposal.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 10 Energy 2 2013-01-01 2013-01-01 false Disposal site design for land disposal. 61.51 Section 61.51 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSING REQUIREMENTS FOR LAND DISPOSAL OF RADIOACTIVE WASTE Technical Requirements for Land Disposal Facilities § 61.51 Disposal site design for...

  12. 24 CFR 3285.103 - Site suitability with design zone maps.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... maps. 3285.103 Section 3285.103 Housing and Urban Development Regulations Relating to Housing and Urban... Considerations § 3285.103 Site suitability with design zone maps. Prior to the initial installation of a new... indicated on the design zone maps provided with the home, are suitable for the site location where the home...

  13. 24 CFR 3285.103 - Site suitability with design zone maps.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... maps. 3285.103 Section 3285.103 Housing and Urban Development Regulations Relating to Housing and Urban... Considerations § 3285.103 Site suitability with design zone maps. Prior to the initial installation of a new... indicated on the design zone maps provided with the home, are suitable for the site location where the home...

  14. 24 CFR 3285.103 - Site suitability with design zone maps.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... maps. 3285.103 Section 3285.103 Housing and Urban Development Regulations Relating to Housing and Urban... Considerations § 3285.103 Site suitability with design zone maps. Prior to the initial installation of a new... indicated on the design zone maps provided with the home, are suitable for the site location where the home...

  15. Fly ash design manual for road and site applications

    SciTech Connect

    DiGioia, A.M. Jr.; Brendel, G.F.

    1992-04-01

    This design manual describes the use of fly ash as a construction material for use as structural and nonstructural fills, backfills, embankments, base courses, roller compacted concrete dams and pavements, soil stabilization, land reclamation and other high volume uses. The manual details the physical, engineering, and chemical properties of bituminous, subbituminous and lignite fly ash. Included are field and laboratory testing methods, design data, procedures and examples, specifications, quality control, and pre- and post-construction monitoring. Volume 1 describes uses where fly ash is used dry or conditioned with small amounts of moisture. Volume 2 describes uses where fly ash is placed as a slurry with relatively large amounts of water.

  16. Design and preparation of a nanoprobe for imaging inflammation sites.

    PubMed

    Yoshitomi, Toru; Nagasaki, Yukio

    2012-12-01

    To image inflammation sites, we developed a novel nanoparticle, hydroxylamine-containing nanoparticle (HANP), which emits an intense electron spin resonance (ESR)-signal triggered by enzymatic oxidation reaction and pH-sensitive self-disintegration. The nanoparticle was prepared from an amphiphilic block copolymer, poly(ethylene glycol)-b-poly[4-(2,2,6,6-tetramethylpiperidine-1-hydroxyl)aminomethylstyrene] (PEG-b-PMNT-H), which spontaneously forms a core-shell type polymeric micelle (particle diameter = ca. 50 nm) in aqueous media. Because the PMNT-H segment in the block copolymer possesses amino groups in each repeating unit, the particle can be disintegrated by protonation of the amino groups in an acidic pH environment such as inflammation sites, which is confined to the hydrophobic core of HANP. Mixing HANP with horseradish peroxidase (HRP)/H(2)O(2) mixture resulted in enzymatic oxidization of the hydroxylamines in the PEG-b-PMNT-H and converted the hydroxylamine to the stable nitroxide radical form in PEG-b-poly[4-(2,2,6,6-tetramethylpiperidine-1-oxyl)aminomethylstyrene] (PEG-b-PMNT), which shows an intense ESR signal. It is interesting to note that the ESR signal increased at a greater rate under acidic conditions (pH 5.6) than that under neutral conditions (pH 7.4), although the enzymatic activity of HRP under neutral conditions is known to be much higher than that under acidic conditions. This indicates that enzymatic oxidation reaction was accelerated by synchronizing the disintegration of HANP under acidic conditions. On the basis of these results, HANP can be used as a high-performance ESR probe for imaging of inflammation sites.

  17. Direct-to-consumer advertising via the Internet: the role of Web site design.

    PubMed

    Sewak, Saurabh S; Wilkin, Noel E; Bentley, John P; Smith, Mickey C

    2005-06-01

    Recent attempts to propose criteria for judging the quality of pharmaceutical and healthcare Web sites do not distinguish between attributes of Web site design related to content and other attributes not related to the content. The Elaboration Likelihood Model from persuasion literature is used as a framework for investigating the effects of Web site design on consequents like attitude and knowledge acquisition. A between-subjects, 2 (high or low involvement)x2 (Web site designed with high or low aspects of visual appeal) factorial design was used in this research. College students were randomly assigned to these treatment groups yielding a balanced design with 29 observations per treatment cell. Analysis of variance results for the effects of involvement and Web site design on attitude and knowledge indicated that the interaction between the independent variables was not significant in both analyses. Examination of main effects revealed that participants who viewed the Web site with higher visual appeal actually had slightly lower knowledge scores (6.32) than those who viewed the Web site with lower visual appeal (7.03, F(1,112)=3.827, P=.053). Results of this research seem to indicate that aspects of Web site design (namely aspects of visual appeal and quality) may not play a role in attaining desired promotional objectives, which can include development of favorable attitudes toward the product and facilitating knowledge acquisition.

  18. The Role of Identity and Culture on Web Site Design

    ERIC Educational Resources Information Center

    Eugene, Wanda; Clark, Kevin

    2009-01-01

    Purpose: The purpose of this paper is to outline an instructional design approach for further development of an African ancestry learning center in order to enhance its educational utility. This is being done in an effort to use technology in transformative ways that extends the practices and opportunities for African Americans.…

  19. HYNOL PROCESS ENGINEERING: PROCESS CONFIGURATION, SITE PLAN, AND EQUIPMENT DESIGN

    EPA Science Inventory

    The report describes the design of the hydropyrolysis reactor system of the Hynol process. (NOTE: A bench scale methanol production facility is being constructed to demonstrate the technical feasibility of producing methanol from biomass using the Hynol process. The plant is bein...

  20. The Role of Identity and Culture on Web Site Design

    ERIC Educational Resources Information Center

    Eugene, Wanda; Clark, Kevin

    2009-01-01

    Purpose: The purpose of this paper is to outline an instructional design approach for further development of an African ancestry learning center in order to enhance its educational utility. This is being done in an effort to use technology in transformative ways that extends the practices and opportunities for African Americans.…

  1. HYNOL PROCESS ENGINEERING: PROCESS CONFIGURATION, SITE PLAN, AND EQUIPMENT DESIGN

    EPA Science Inventory

    The report describes the design of the hydropyrolysis reactor system of the Hynol process. (NOTE: A bench scale methanol production facility is being constructed to demonstrate the technical feasibility of producing methanol from biomass using the Hynol process. The plant is bein...

  2. Site study plan for EDBH (Engineering Design Boreholes) seismic surveys, Deaf Smith County site, Texas: Revision 1

    SciTech Connect

    Hume, H.

    1987-12-01

    This site study plan describes seismic reflection surveys to run north-south and east-west across the Deaf Smith County site, and intersecting near the Engineering Design Boreholes (EDBH). Both conventional and shallow high-resolution surveys will be run. The field program has been designed to acquire subsurface geologic and stratigraphic data to address information/data needs resulting from Federal and State regulations and Repository program requirements. The data acquired by the conventional surveys will be common-depth- point, seismic reflection data optimized for reflection events that indicate geologic structure near the repository horizon. The data will also resolve the basement structure and shallow reflection events up to about the top of the evaporite sequence. Field acquisition includes a testing phase to check/select parameters and a production phase. The field data will be subjected immediately to conventional data processing and interpretation to determine if there are any anamolous structural for stratigraphic conditions that could affect the choice of the EDBH sites. After the EDBH's have been drilled and logged, including vertical seismic profiling, the data will be reprocessed and reinterpreted for detailed structural and stratigraphic information to guide shaft development. The shallow high-resulition seismic reflection lines will be run along the same alignments, but the lines will be shorter and limited to immediate vicinity of the EDBH sites. These lines are planned to detect faults or thick channel sands that may be present at the EDBH sites. 23 refs. , 7 figs., 5 tabs.

  3. SDM-Assist software to design site-directed mutagenesis primers introducing "silent" restriction sites.

    PubMed

    Karnik, Abhijit; Karnik, Rucha; Grefen, Christopher

    2013-03-22

    Over the past decades site-directed mutagenesis (SDM) has become an indispensable tool for biological structure-function studies. In principle, SDM uses modified primer pairs in a PCR reaction to introduce a mutation in a cDNA insert. DpnI digestion of the reaction mixture is used to eliminate template copies before amplification in E. coli; however, this process is inefficient resulting in un-mutated clones which can only be distinguished from mutant clones by sequencing. We have developed a program - 'SDM-Assist' which creates SDM primers adding a specific identifier: through additional silent mutations a restriction site is included or a previous one removed which allows for highly efficient identification of 'mutated clones' by a simple restriction digest. The direct identification of SDM clones will save time and money for researchers. SDM-Assist also scores the primers based on factors such as Tm, GC content and secondary structure allowing for simplified selection of optimal primer pairs.

  4. REMOT: A Design for Multiple Site Remote Observing

    NASA Astrophysics Data System (ADS)

    Linde, P.; Pasian, F.; Pucillo, M.; Ponz, J. D.

    The REMOT project objective is to develop and validate a generic approach to allow remote control of scientific experiments and facilities that require real time operation and multimedia information feedback. The project is funded through the European Union Telematics initiative and is a collaboration, involving representatives from both the astro- and plasma physics communities. Standard communications infrastructure and software solutions will be used wherever possible. In the first step, requirements have been collected and analysed, resulting in a set of service definitions. These have been partly implemented to perform a set of demonstrations, showing the feasibility of the design.

  5. Rosetta and the Design of Ligand Binding Sites.

    PubMed

    Moretti, Rocco; Bender, Brian J; Allison, Brittany; Meiler, Jens

    2016-01-01

    Proteins that bind small molecules (ligands) can be used as biosensors, signal modulators, and sequestering agents. When naturally occurring proteins for a particular target ligand are not available, artificial proteins can be computationally designed. We present a protocol based on RosettaLigand to redesign an existing protein pocket to bind a target ligand. Starting with a protein structure and the structure of the ligand, Rosetta can optimize both the placement of the ligand in the pocket and the identity and conformation of the surrounding sidechains, yielding proteins that bind the target compound.

  6. 41 CFR 102-76.20 - What issues must Federal agencies consider in providing site planning and landscape design services?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... agencies consider in providing site planning and landscape design services? 102-76.20 Section 102-76.20... What issues must Federal agencies consider in providing site planning and landscape design services? In providing site planning and design services, Federal agencies must— (a) Make the site planning and landscape...

  7. 41 CFR 102-76.20 - What issues must Federal agencies consider in providing site planning and landscape design services?

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... agencies consider in providing site planning and landscape design services? 102-76.20 Section 102-76.20... What issues must Federal agencies consider in providing site planning and landscape design services? In providing site planning and design services, Federal agencies must— (a) Make the site planning and landscape...

  8. Nuclear depletion of apurinic/apyrimidinic endonuclease 1 (Ape1/Ref-1) is an indicator of energy disruption in neurons.

    PubMed

    Singh, Shilpee; Englander, Ella W

    2012-11-01

    Apurinic/apyrimidinic endonuclease 1 (Ape1/Ref-1) is a multifunctional protein critical for cellular survival. Its involvement in adaptive survival responses includes key roles in redox sensing, transcriptional regulation, and repair of DNA damage via the base excision repair (BER) pathway. Ape1 is abundant in most cell types and central in integrating the first BER step catalyzed by different DNA glycosylases. BER is the main process for removal of oxidative DNA lesions in postmitotic brain cells, and after ischemic brain injury preservation of Ape1 coincides with neuronal survival, while its loss has been associated with neuronal death. Here, we report that in cultured primary neurons, diminution of cellular ATP by either oligomycin or H(2)O(2) is accompanied by depletion of nuclear Ape1, while other BER proteins are unaffected and retain their nuclear localization under these conditions. Importantly, while H(2)O(2) induces γH2AX phosphorylation, indicative of chromatin rearrangements in response to DNA damage, oligomycin does not. Furthermore, despite comparable diminution of ATP content, H(2)O(2) and oligomycin differentially affect critical parameters of mitochondrial respiration that ultimately determine cellular ATP content. Taken together, our findings demonstrate that in neurons, nuclear compartmentalization of Ape1 depends on ATP and loss of nuclear Ape1 reflects disruption of neuronal energy homeostasis. Energy crisis is a hallmark of stroke and other ischemic/hypoxic brain injuries. In vivo studies have shown that Ape1 deficit precedes neuronal loss in injured brain regions. Thus, our findings bring to light the possibility that energy failure-induced Ape1 depletion triggers neuronal death in ischemic brain injuries.

  9. Apurinic/Apyrimidinic Endonuclease/Redox Factor-1 (APE1/Ref-1) Redox Function Negatively Regulates NRF2*

    PubMed Central

    Fishel, Melissa L.; Wu, Xue; Devlin, Cecilia M.; Logsdon, Derek P.; Jiang, Yanlin; Luo, Meihua; He, Ying; Yu, Zhangsheng; Tong, Yan; Lipking, Kelsey P.; Maitra, Anirban; Rajeshkumar, N. V.; Scandura, Glenda; Kelley, Mark R.; Ivan, Mircea

    2015-01-01

    Apurinic/apyrimidinic endonuclease/redox factor-1 (APE1/Ref-1) (henceforth referred to as Ref-1) is a multifunctional protein that in addition to its base excision DNA repair activity exerts redox control of multiple transcription factors, including nuclear factor κ-light chain enhancer of activated B cells (NF-κB), STAT3, activator protein-1 (AP-1), hypoxia-inducible factor-1 (HIF-1), and tumor protein 53 (p53). In recent years, Ref-1 has emerged as a promising therapeutic target in cancer, particularly in pancreatic ductal carcinoma. Although a significant amount of research has centered on Ref-1, no wide-ranging approach had been performed on the effects of Ref-1 inhibition and transcription factor activity perturbation. Starting with a broader approach, we identified a previously unsuspected effect on the nuclear factor erythroid-related factor 2 (NRF2), a critical regulator of cellular defenses against oxidative stress. Based on genetic and small molecule inhibitor-based methodologies, we demonstrated that repression of Ref-1 potently activates NRF2 and its downstream targets in a dose-dependent fashion, and that the redox, rather than the DNA repair function of Ref-1 is critical for this effect. Intriguingly, our results also indicate that this pathway does not involve reactive oxygen species. The link between Ref-1 and NRF2 appears to be present in all cells tested in vitro, noncancerous and cancerous, including patient-derived tumor samples. In particular, we focused on understanding the implications of the novel interaction between these two pathways in primary pancreatic ductal adenocarcinoma tumor cells and provide the first evidence that this mechanism has implications for overcoming the resistance against experimental drugs targeting Ref-1 activity, with clear translational implications. PMID:25492865

  10. Inhibitors of the apurinic/apyrimidinic endonuclease 1 (APE1)/nucleophosmin (NPM1) interaction that display anti-tumor properties.

    PubMed

    Poletto, Mattia; Malfatti, Matilde C; Dorjsuren, Dorjbal; Scognamiglio, Pasqualina L; Marasco, Daniela; Vascotto, Carlo; Jadhav, Ajit; Maloney, David J; Wilson, David M; Simeonov, Anton; Tell, Gianluca

    2016-05-01

    The apurinic/apyrimidinic endonuclease 1 (APE1) is a protein central to the base excision DNA repair pathway and operates in the modulation of gene expression through redox-dependent and independent mechanisms. Aberrant expression and localization of APE1 in tumors are recurrent hallmarks of aggressiveness and resistance to therapy. We identified and characterized the molecular association between APE1 and nucleophosmin (NPM1), a multifunctional protein involved in the preservation of genome stability and rRNA maturation. This protein-protein interaction modulates subcellular localization and endonuclease activity of APE1. Moreover, we reported a correlation between APE1 and NPM1 expression levels in ovarian cancer, with NPM1 overexpression being a marker of poor prognosis. These observations suggest that tumors that display an augmented APE1/NPM1 association may exhibit increased aggressiveness and resistance. Therefore, targeting the APE1/NPM1 interaction might represent an innovative strategy for the development of anticancer drugs, as tumor cells relying on higher levels of APE1 and NPM1 for proliferation and survival may be more sensitive than untransformed cells. We set up a chemiluminescence-based high-throughput screening assay in order to find small molecules able to interfere with the APE1/NPM1 interaction. This screening led to the identification of a set of bioactive compounds that impair the APE1/NPM1 association in living cells. Interestingly, some of these molecules display anti-proliferative activity and sensitize cells to therapeutically relevant genotoxins. Given the prognostic significance of APE1 and NPM1, these compounds might prove effective in the treatment of tumors that show abundant levels of both proteins, such as ovarian or hepatic carcinomas. © 2015 Wiley Periodicals, Inc.

  11. Does implant design affect primary stability in extraction sites?

    PubMed

    Karl, Matthias; Irastorza-Landa, Ainara

    2017-01-01

    Immediate implant placement and restoration following tooth extraction is desirable for shortening treatment times and maintaining oral structures. With a variety of bone-level implants available on the market, the goal of this in-vitro study was to compare different implant designs. Using a polyurethane foam sandwich model with an intermediate cortical layer, a standardized extraction socket model was created. The following implant systems were assessed: OsseoSpeed EV (Astra), Straumann Bone Level Tapered (BLT), and Nobel Active (NA). Implant stability assessment included implant insertion torque (IT) and implant stability quotient (ISQ). The three implant systems were compared in a statistical analysis based on the two sample tests with the level of significance set at α = .05 and Bonferroni correction for multiple comparisons. The relationship between IT and ISQ was assessed using the Pearson correlation coefficient. IT values were highest for NA implants, with a mean of 36.52 Ncm, which was significantly greater than in BLT (P < .001) and Astra (P < .001). ISQ values were highest for NA, with a mean 53.9, which was significantly higher than for BLT (P < .001) but not for Astra (P = .07). BLT showed significantly lower ISQ as compared to NA (P < .001) and Astra (P < .001). Overall, the IT and ISQ measurements did not correlate. The developed extraction model allowed for consistent measurements of implant stability parameters at a clinically relevant level. Implant design seems to play a relevant role for achieving primary stability in challenging situations. Next evaluation steps should involve testing under dynamic loading conditions.

  12. Fracture analysis and rock quality designation estimation for the Yucca Mountain Site Characterization Project; Yucca Mountain Site Characterization Project

    SciTech Connect

    Lin, M.; Hardy, M.P.; Bauer, S.J.

    1993-02-01

    Within the Yucca Mountain Site Characterization Project, the design of drifts and ramps and evaluation of the impacts of thermomechanical loading of the host rock requires definition of the rock mass mechanical properties. Ramps and exploratory drifts will intersect both welded and nonwelded tuffs with varying abundance of fractures. The rock mass mechanical properties are dependent on the intact rock properties and the fracture joint characteristics. An understanding of the effects of fractures on the mechanical properties of the rock mass begins with a detailed description of the fracture spatial location and abundance, and includes a description of their physical characteristics. This report presents a description of the abundance, orientation, and physical characteristics of fractures and the Rock Quality Designation in the thermomechanical stratigraphic units at the Yucca Mountain site. Data was reviewed from existing sources and used to develop descriptions for each unit. The product of this report is a data set of the best available information on the fracture characteristics.

  13. Comment and response document for the final remedial action plan and site design for stabilization of the inactive uranium mill tailings sites at Slick Rock, Colorado. Revision 2

    SciTech Connect

    1996-05-01

    This document for the final remedial action plan and site design has been prepared for US Department of Energy Environmental Restoration Division as part of the Uranium Mill Tailings Remedial Action plan. Comments and responses are included for the site design for stabilization of the inactive uranium mill tailings sites at Slick Rock, Colorado.

  14. Title I conceptual design for Pit 6 landfill closure at Lawrence Livermore National Laboratory Site 300

    SciTech Connect

    MacDonnell, B.A.; Obenauf, K.S.

    1996-08-01

    The objective of this design project is to evaluate and prepare design and construction documents for a closure cover cap for the Pit 6 Landfill located at Lawrence Livermore National Laboratory Site 300. This submittal constitutes the Title I Design (Conceptual Design) for the closure cover of the Pit 6 Landfill. A Title I Design is generally 30 percent of the design effort. Title H Design takes the design to 100 percent complete. Comments and edits to this Title I Design will be addressed in the Title II design submittal. Contents of this report are as follows: project background; design issues and engineering approach; design drawings; calculation packages; construction specifications outline; and construction quality assurance plan outline.

  15. NGA-West 2 GMPE average site coefficients for use in earthquake-resistant design

    USGS Publications Warehouse

    Borcherdt, Roger D.

    2015-01-01

    Site coefficients corresponding to those in tables 11.4–1 and 11.4–2 of Minimum Design Loads for Buildings and Other Structures published by the American Society of Civil Engineers (Standard ASCE/SEI 7-10) are derived from four of the Next Generation Attenuation West2 (NGA-W2) Ground-Motion Prediction Equations (GMPEs). The resulting coefficients are compared with those derived by other researchers and those derived from the NGA-West1 database. The derivation of the NGA-W2 average site coefficients provides a simple procedure to update site coefficients with each update in the Maximum Considered Earthquake Response MCER maps. The simple procedure yields average site coefficients consistent with those derived for site-specific design purposes. The NGA-W2 GMPEs provide simple scale factors to reduce conservatism in current simplified design procedures.

  16. Identification and Characterization of Inhibitors of Human Apurinic/apyrimidinic Endonuclease APE1

    PubMed Central

    Simeonov, Anton; Kulkarni, Avanti; Dorjsuren, Dorjbal; Jadhav, Ajit; Shen, Min; McNeill, Daniel R.; Austin, Christopher P.; Wilson, David M.

    2009-01-01

    APE1 is the major nuclease for excising abasic (AP) sites and particular 3′-obstructive termini from DNA, and is an integral participant in the base excision repair (BER) pathway. BER capacity plays a prominent role in dictating responsiveness to agents that generate oxidative or alkylation DNA damage, as well as certain chain-terminating nucleoside analogs and 5-fluorouracil. We describe within the development of a robust, 1536-well automated screening assay that employs a deoxyoligonucleotide substrate operating in the red-shifted fluorescence spectral region to identify APE1 endonuclease inhibitors. This AP site incision assay was used in a titration-based high-throughput screen of the Library of Pharmacologically Active Compounds (LOPAC1280), a collection of well-characterized, drug-like molecules representing all major target classes. Prioritized hits were authenticated and characterized via two high-throughput screening assays – a Thiazole Orange fluorophore-DNA displacement test and an E. coli endonuclease IV counterscreen – and a conventional, gel-based radiotracer incision assay. The top, validated compounds, i.e. 6-hydroxy-DL-DOPA, Reactive Blue 2 and myricetin, were shown to inhibit AP site cleavage activity of whole cell protein extracts from HEK 293T and HeLa cell lines, and to enhance the cytotoxic and genotoxic potency of the alkylating agent methylmethane sulfonate. The studies herein report on the identification of novel, small molecule APE1-targeted bioactive inhibitor probes, which represent initial chemotypes towards the development of potential pharmaceuticals. PMID:19484131

  17. World Wide Web Sites for Instructional Design and Human Performance Technology.

    ERIC Educational Resources Information Center

    Loughner, Pamela; Milheim, William D.

    1999-01-01

    Describes different Internet resource sites for instructional design and human performance technology maintained by various groups including: professional organizations, United States government agencies, educational institutions, publishers and booksellers, and miscellaneous groups. (AEF)

  18. Site Planning and Design to Enable Planetary Science and Human Exploration

    NASA Astrophysics Data System (ADS)

    Lewis, R.; Lupisella, M.; Bleacher, J.

    2017-02-01

    It is critical to properly plan site layout and design of science and habitation assets about and within planetary exploration zones following planetary protection policies and environment management practices for effective robotic and human missions.

  19. 40 CFR 228.15 - Dumping sites designated on a final basis.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... materials. Designation of these sites was based on environmental studies conducted in accordance with the... for any ongoing long-term studies), including the identification of alternatives to open-water...

  20. 40 CFR 228.15 - Dumping sites designated on a final basis.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... materials. Designation of these sites was based on environmental studies conducted in accordance with the... for any ongoing long-term studies), including the identification of alternatives to open-water...

  1. 40 CFR 228.15 - Dumping sites designated on a final basis.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... materials. Designation of these sites was based on environmental studies conducted in accordance with the... for any ongoing long-term studies), including the identification of alternatives to open-water...

  2. A user-centered model for web site design: needs assessment, user interface design, and rapid prototyping.

    PubMed

    Kinzie, Mable B; Cohn, Wendy F; Julian, Marti F; Knaus, William A

    2002-01-01

    As the Internet continues to grow as a delivery medium for health information, the design of effective Web sites becomes increasingly important. In this paper, the authors provide an overview of one effective model for Web site design, a user-centered process that includes techniques for needs assessment, goal/task analysis, user interface design, and rapid prototyping. They detail how this approach was employed to design a family health history Web site, Health Heritage . This Web site helps patients record and maintain their family health histories in a secure, confidential manner. It also supports primary care physicians through analysis of health histories, identification of potential risks, and provision of health care recommendations. Visual examples of the design process are provided to show how the use of this model resulted in an easy-to-use Web site that is likely to meet user needs. The model is effective across diverse content arenas and is appropriate for applications in varied media.

  3. Facilitating Participation: From the EML Web Site to the Learning Network for Learning Design

    ERIC Educational Resources Information Center

    Hummel, Hans G. K.; Tattersall, Colin; Burgos, Daniel; Brouns, Francis; Kurvers, Hub; Koper, Rob

    2005-01-01

    This article investigates conditions for increasing active participation in on-line communities. As a case study, we use three generations of facilities designed to promote learning in the area of Educational Modelling Languages. Following a description of early experience with a conventional web site and with a community site offering facilities…

  4. The Role of Virtual Reference in Library Web Site Design: A Qualitative Source for Usage Data

    ERIC Educational Resources Information Center

    Powers, Amanda Clay; Shedd, Julie; Hill, Clay

    2011-01-01

    Gathering qualitative information about usage behavior of library Web sites is a time-consuming process requiring the active participation of patron communities. Libraries that collect virtual reference transcripts, however, hold valuable data regarding how the library Web site is used that could benefit Web designers. An analysis of virtual…

  5. Designing a Web Site for One Imaginary Persona that Reflect the Needs of Many

    ERIC Educational Resources Information Center

    Cunningham, Heather

    2005-01-01

    Does designing a large academic library Web site for an imaginary person seem a sensible thing to do? As a reference librarian and member of the Web team of the Gerstein Science Information Centre (Gerstein Library), that is exactly what this author set out to do in May 2005. She proposed that the best way to ensure that the Web site meets the…

  6. Sharing Designer and User Perspectives of Web Site Evaluation: A Cross-Campus Collaborative Learning Experience.

    ERIC Educational Resources Information Center

    Collings, Penny; Pearce, Jon

    2002-01-01

    Presents an online collaborative process that facilitates usability evaluation of Web sites. Describes how the system was designed and used by students and staff at two Australian universities and shows that the process provides feedback on Web site usability and the experience of usability evaluation from the perspectives of a user and a…

  7. The Path to Graduation: A Model Interactive Web Site Design Supporting Doctoral Students

    ERIC Educational Resources Information Center

    Simmons-Johnson, Nicole

    2012-01-01

    Objective. This 2-phase mixed method study assessed 2nd-year doctoral students' and dissertation students' perceptions of the current Graduate School of Education dissertation support Web site, with implications for designing a model dissertation support Web site. Methods. Phase 1 collected quantitative and qualitative data through an…

  8. Design Insights and Inspiration from the Tate: What Museum Web Sites Can Offer Us

    ERIC Educational Resources Information Center

    Riley-Huff, Debra A.

    2009-01-01

    There are many similarities between museums and academic libraries as public service institutions. This article is an examination of museum Web site practices and concepts that might also be transferable to academic library Web sites. It explores the digital manifestations of design and information presentation, user engagement, interactivity, and…

  9. A Checklist for Designing and Evaluating Physical Education Program Web Sites

    ERIC Educational Resources Information Center

    Tucker, Michael; Hill, Grant

    2009-01-01

    Creating a physical education department web site is an excellent way to promote a positive image of the program, because students and parents are able to find important information and improve the lines of communication. A well-designed physical education web site can even encourage students to increase their physical activity levels. Improved…

  10. The Path to Graduation: A Model Interactive Web Site Design Supporting Doctoral Students

    ERIC Educational Resources Information Center

    Simmons-Johnson, Nicole

    2012-01-01

    Objective. This 2-phase mixed method study assessed 2nd-year doctoral students' and dissertation students' perceptions of the current Graduate School of Education dissertation support Web site, with implications for designing a model dissertation support Web site. Methods. Phase 1 collected quantitative and qualitative data through an…

  11. The Role of Virtual Reference in Library Web Site Design: A Qualitative Source for Usage Data

    ERIC Educational Resources Information Center

    Powers, Amanda Clay; Shedd, Julie; Hill, Clay

    2011-01-01

    Gathering qualitative information about usage behavior of library Web sites is a time-consuming process requiring the active participation of patron communities. Libraries that collect virtual reference transcripts, however, hold valuable data regarding how the library Web site is used that could benefit Web designers. An analysis of virtual…

  12. Design Insights and Inspiration from the Tate: What Museum Web Sites Can Offer Us

    ERIC Educational Resources Information Center

    Riley-Huff, Debra A.

    2009-01-01

    There are many similarities between museums and academic libraries as public service institutions. This article is an examination of museum Web site practices and concepts that might also be transferable to academic library Web sites. It explores the digital manifestations of design and information presentation, user engagement, interactivity, and…

  13. A Checklist for Designing and Evaluating Physical Education Program Web Sites

    ERIC Educational Resources Information Center

    Tucker, Michael; Hill, Grant

    2009-01-01

    Creating a physical education department web site is an excellent way to promote a positive image of the program, because students and parents are able to find important information and improve the lines of communication. A well-designed physical education web site can even encourage students to increase their physical activity levels. Improved…

  14. 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

  15. Nevada test site low-level and mixed waste repository design in the unsaturated zone

    SciTech Connect

    Kawamura, T.A.; Warren, D.M.; USDOE Nevada Operations Office, Las Vegas, NV )

    1989-01-01

    The Area 5 Radioactive Waste Management Site (RWMS) at the Nevada Test Site (NTS) is used for shallow land disposal of Low-Level Radioactive (LLW) and for retrievable disposal of Mixed Wastes (MW) from various Department of Energy (DOE) facilities. The site is situated in southern Nevada, one of the most arid regions of the United States. Design considerations include vadose zone monitoring in lieu of groundwater monitoring, stringent waste acceptance and packaging criteria, a waste examination and real-time radiography facility, and trench design. 4 refs.

  16. Remedial action plan and site design for stabilization of the inactive uranium mill tailings sites at Rifle, Colorado. Volume 2, Appendices D and E: Final report

    SciTech Connect

    Not Available

    1992-02-01

    This appendix assesses the present conditions and data gathered about the two inactive uranium mill tailings sites near Rifle, Colorado, and the designated disposal site six miles north of Rifle in the area of Estes Gulch. It consolidates available engineering, radiological, geotechnical, hydrological, meteorological, and other information pertinent to the design of the Remedial Action Plan (RAP). The data characterize conditions at the mill, tailings, and disposal site so that the Remedial Action Contractor (RAC) may complete final designs for the remedial actions.

  17. A usability evaluation exploring the design of American Nurses Association state web sites.

    PubMed

    Alexander, Gregory L; Wakefield, Bonnie J; Anbari, Allison B; Lyons, Vanessa; Prentice, Donna; Shepherd, Marilyn; Strecker, E Bradley; Weston, Marla J

    2014-08-01

    National leaders are calling for opportunities to facilitate the Future of Nursing. Opportunities can be encouraged through state nurses association Web sites, which are part of the American Nurses Association, that are well designed, with appropriate content, and in a language professional nurses understand. The American Nurses Association and constituent state nurses associations provide information about nursing practice, ethics, credentialing, and health on Web sites. We conducted usability evaluations to determine compliance with heuristic and ethical principles for Web site design. We purposefully sampled 27 nursing association Web sites and used 68 heuristic and ethical criteria to perform systematic usability assessments of nurse association Web sites. Web site analysis included seven double experts who were all RNs trained in usability analysis. The extent to which heuristic and ethical criteria were met ranged widely from one state that met 0% of the criteria for "help and documentation" to states that met greater than 92% of criteria for "visibility of system status" and "aesthetic and minimalist design." Suggested improvements are simple yet make an impact on a first-time visitor's impression of the Web site. For example, adding internal navigation and tracking features and providing more details about the application process through help and frequently asked question documentation would facilitate better use. Improved usability will improve effectiveness, efficiency, and consumer satisfaction with these Web sites.

  18. 10 CFR 433.6 - Sustainable principles for siting, design and construction. [Reserved

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 10 Energy 3 2011-01-01 2011-01-01 false Sustainable principles for siting, design and construction. 433.6 Section 433.6 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY STANDARDS FOR THE DESIGN AND CONSTRUCTION OF NEW FEDERAL COMMERCIAL AND MULTI-FAMILY HIGH-RISE RESIDENTIAL BUILDINGS...

  19. 10 CFR 433.6 - Sustainable principles for siting, design and construction. [Reserved

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 10 Energy 3 2010-01-01 2010-01-01 false Sustainable principles for siting, design and construction. 433.6 Section 433.6 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY STANDARDS FOR THE DESIGN AND CONSTRUCTION OF NEW FEDERAL COMMERCIAL AND MULTI-FAMILY HIGH-RISE RESIDENTIAL BUILDINGS...

  20. Interfacing Media: User-Centered Design for Media-Rich Web Sites.

    ERIC Educational Resources Information Center

    Horton, Sarah

    2000-01-01

    Discusses multimedia Web site design that may include images, animations, audio, and video. Highlights include interfaces that stress user-centered design; using only relevant media; placing high-demand content on secondary pages and keeping the home page simpler; providing information about the media; considering users with disabilities; and user…

  1. Interfacing Media: User-Centered Design for Media-Rich Web Sites.

    ERIC Educational Resources Information Center

    Horton, Sarah

    2000-01-01

    Discusses multimedia Web site design that may include images, animations, audio, and video. Highlights include interfaces that stress user-centered design; using only relevant media; placing high-demand content on secondary pages and keeping the home page simpler; providing information about the media; considering users with disabilities; and user…

  2. Computational Tools for Allosteric Drug Discovery: Site Identification and Focus Library Design.

    PubMed

    Huang, Wenkang; Nussinov, Ruth; Zhang, Jian

    2017-01-01

    Allostery is an intrinsic phenomenon of biological macromolecules involving regulation and/or signal transduction induced by a ligand binding to an allosteric site distinct from a molecule's active site. Allosteric drugs are currently receiving increased attention in drug discovery because drugs that target allosteric sites can provide important advantages over the corresponding orthosteric drugs including specific subtype selectivity within receptor families. Consequently, targeting allosteric sites, instead of orthosteric sites, can reduce drug-related side effects and toxicity. On the down side, allosteric drug discovery can be more challenging than traditional orthosteric drug discovery due to difficulties associated with determining the locations of allosteric sites and designing drugs based on these sites and the need for the allosteric effects to propagate through the structure, reach the ligand binding site and elicit a conformational change. In this study, we present computational tools ranging from the identification of potential allosteric sites to the design of "allosteric-like" modulator libraries. These tools may be particularly useful for allosteric drug discovery.

  3. Prognostic Significance of Human Apurinic/Apyrimidinic Endonuclease (APE/Ref-1) Expression in Rectal Cancer Treated With Preoperative Radiochemotherapy

    SciTech Connect

    Kim, Jun-Sang; Kim, Jin-Man; Liang, Zhe Long; Jang, Ji Young; Kim, Sup; Huh, Gil Ja; Kim, Ki-Hwan; Cho, Moon-June

    2012-01-01

    Purpose: Human apurinic endonuclease/redox factor 1 (APE/Ref-1) mediates repair of radiation-induced DNA lesions and regulates transcription via redox-based activation. We investigated the predictive and prognostic significance of APE/Ref-1 expression in pretreatment biopsy specimens in locally advanced rectal cancer (LARC) (cT3-T4 or N+). Methods and Materials: APE/Ref-1 expression was analyzed by immunohistochemistry in pretreatment biopsy specimens obtained from 83 patients with LARC. Patients received preoperative radiotherapy of 50.4 Gy in 28 fractions, combined with oral capecitabine and leucovorin chemotherapy, followed by curative surgery. The prognostic significance of various clinicopathologic characteristics, including APE/Ref-1 protein expression, was evaluated. Results: APE/Ref-1 was expressed in 97% of patient samples. Exclusive APE/Ref-1 nuclear staining was observed in 49 of 83 samples (59%), and mixed nuclear and cytoplasmic staining was observed in 31 samples (37%). APE/Ref-1 nuclear expression levels were low in 49 patients (59%) and high in 34 patients (41%). The level of APE/Ref-1 nuclear expression was not a prognostic factor for overall and disease-free survival. Cytoplasmic expression of APE/Ref-1 was a borderline-significant predictive factor for pathologic tumor response (p = 0.08) and a significant prognostic factor for disease-free survival, as shown by univariate analysis (p = 0.037). Multivariate analysis confirmed that cytoplasmic localization of APE/Ref-1 is a significant predictor of disease-free survival (hazard ratio, 0.45; p = 0.046). Conclusions: APE/Ref-1 was expressed in a majority of pretreatment biopsy specimens from patients with LARC. The level of APE/Ref-1 nuclear expression was not a significant predictive and prognostic factor; however, cytoplasmic localization of the protein was negatively associated with disease-free survival. These results indicate that cytoplasmic expression of APE/Ref-1 represents an adverse

  4. National Ignition Facility subsystem design requirements NIF site improvements SSDR 1.2.1

    SciTech Connect

    Kempel, P.; Hands, J.

    1996-08-19

    This Subsystem Design Requirements (SSDR) document establishes the performance, design, and verification requirements associated with the NIF Project Site at Lawrence Livermore National Laboratory (LLNL) at Livermore, California. It identifies generic design conditions for all NIF Project facilities, including siting requirements associated with natural phenomena, and contains specific requirements for furnishing site-related infrastructure utilities and services to the NIF Project conventional facilities and experimental hardware systems. Three candidate sites were identified as potential locations for the NIF Project. However, LLNL has been identified by DOE as the preferred site because of closely related laser experimentation underway at LLNL, the ability to use existing interrelated infrastructure, and other reasons. Selection of a site other than LLNL will entail the acquisition of site improvements and infrastructure additional to those described in this document. This SSDR addresses only the improvements associated with the NIF Project site located at LLNL, including new work and relocation or demolition of existing facilities that interfere with the construction of new facilities. If the Record of Decision for the PEIS on Stockpile Stewardship and Management were to select another site, this SSDR would be revised to reflect the characteristics of the selected site. Other facilities and infrastructure needed to support operation of the NIF, such as those listed below, are existing and available at the LLNL site, and are not included in this SSDR. Office Building. Target Receiving and Inspection. General Assembly Building. Electro- Mechanical Shop. Warehousing and General Storage. Shipping and Receiving. General Stores. Medical Facilities. Cafeteria services. Service Station and Garage. Fire Station. Security and Badging Services.

  5. Linkage between site characterization program and performance and design data needs

    SciTech Connect

    Blanchard, M.B.; Dobson, D.C.; Younker, J.L.; Voegele, M.D.

    1990-04-01

    The basis for identifying site-specific data needs for the Yucca Mountain site was a thorough review and evaluation of the regulatory requirements for a geologic repository. The US Nuclear Regulatory Commission`s (NRC) regulation governing disposal of high-level waste in a geologic repository utilizes a multiple barrier approach that includes reliance on both engineered and natural elements. Given an understanding of the regulatory requirements, the next step was to identify the natural features of the site and the components of the engineered barriers considered most likely to make significant contributions to isolation of waste at Yucca Mountain. The intent was to develop a site testing program that focused on understanding those natural site characteristics and processes most likely to contribute to waste isolation; and providing site data necessary to design and assess the performance of engineered barriers. 3 refs., 3 figs.

  6. Comparison of SARS and NL63 papain-like protease binding sites and binding site dynamics: inhibitor design implications.

    PubMed

    Chaudhuri, Rima; Tang, Sishi; Zhao, Guijun; Lu, Hui; Case, David A; Johnson, Michael E

    2011-11-25

    The human severe acute respiratory syndrome coronavirus (SARS-CoV) and the NL63 coronaviruses are human respiratory pathogens for which no effective antiviral treatment exists. The papain-like cysteine proteases encoded by the coronavirus (SARS-CoV: PLpro; NL63: PLP1 and PLP2) represent potential targets for antiviral drug development. Three recent inhibitor-bound PLpro structures highlight the role of an extremely flexible six-residue loop in inhibitor binding. The high binding site plasticity is a major challenge in computational drug discovery/design efforts. From conventional molecular dynamics and accelerated molecular dynamics (aMD) simulations, we find that with conventional molecular dynamics simulation, PLpro translationally samples the open and closed conformation of BL2 loop on a picosecond-nanosecond timescale but does not reproduce the peptide bond inversion between loop residues Tyr269 and Gln270 that is observed on inhibitor GRL0617 binding. Only aMD simulation, starting from the closed loop conformation, reproduced the 180° ϕ-ψ dihedral rotation back to the open loop state. The Tyr-Gln peptide bond inversion appears to involve a progressive conformational change of the full loop, starting at one side, and progressing to the other. We used the SARS-CoV apo X-ray structure to develop a model of the NL63-PLP2 catalytic site. Superimposition of the PLP2 model on the PLpro X-ray structure identifies binding site residues in PLP2 that contribute to the distinct substrate cleavage site specificities between the two proteases. The topological and electrostatic differences between the two protease binding sites also help explain the selectivity of non-covalent PLpro inhibitors.

  7. Study of seismic design bases and site conditions for nuclear power plants

    SciTech Connect

    Not Available

    1980-04-01

    This report presents the results of an investigation of four topics pertinent to the seismic design of nuclear power plants: Design accelerations by regions of the continental United States; review and compilation of design-basis seismic levels and soil conditions for existing nuclear power plants; regional distribution of shear wave velocity of foundation materials at nuclear power plant sites; and technical review of surface-founded seismic analysis versus embedded approaches.

  8. Usage and design evaluation by family caregivers of a stroke intervention web site.

    PubMed

    Pierce, Linda L; Steiner, Victoria

    2013-10-01

    Four of five families are affected by stroke. Many caregivers access the Internet and gather healthcare information from Web-based sources. The purpose of this descriptive evaluation was to assess the usage and design of the Caring∼Web site, which provides education/support for family caregivers of persons with stroke residing in home settings. Thirty-six caregivers from two Midwest states accessed this intervention in a 1-year study. The average participant was 54 years old, White, woman, and the spouse of the care recipient. In a telephone interview, four Web site questions were asked twice a month/bimonthly, and a 33-item survey at the conclusion of the study evaluated the Web site usage and design of its components. Descriptive analysis methods were used, and statistics were collected on the number of visits to the Web site. On average, participants logged on to the Web site 1-2 hours per week, although usage declined after several months for some participants. Participants positively rated the Web site's appearance and usability that included finding the training to be adequate. Web site designers can replicate this intervention for other health conditions.

  9. High-Probability Neurotransmitter Release Sites Represent an Energy-Efficient Design.

    PubMed

    Lu, Zhongmin; Chouhan, Amit K; Borycz, Jolanta A; Lu, Zhiyuan; Rossano, Adam J; Brain, Keith L; Zhou, You; Meinertzhagen, Ian A; Macleod, Gregory T

    2016-10-10

    Nerve terminals contain multiple sites specialized for the release of neurotransmitters. Release usually occurs with low probability, a design thought to confer many advantages. High-probability release sites are not uncommon, but their advantages are not well understood. Here, we test the hypothesis that high-probability release sites represent an energy-efficient design. We examined release site probabilities and energy efficiency at the terminals of two glutamatergic motor neurons synapsing on the same muscle fiber in Drosophila larvae. Through electrophysiological and ultrastructural measurements, we calculated release site probabilities to differ considerably between terminals (0.33 versus 0.11). We estimated the energy required to release and recycle glutamate from the same measurements. The energy required to remove calcium and sodium ions subsequent to nerve excitation was estimated through microfluorimetric and morphological measurements. We calculated energy efficiency as the number of glutamate molecules released per ATP molecule hydrolyzed, and high-probability release site terminals were found to be more efficient (0.13 versus 0.06). Our analytical model indicates that energy efficiency is optimal (∼0.15) at high release site probabilities (∼0.76). As limitations in energy supply constrain neural function, high-probability release sites might ameliorate such constraints by demanding less energy. Energy efficiency can be viewed as one aspect of nerve terminal function, in balance with others, because high-efficiency terminals depress significantly during episodic bursts of activity. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Probing Enhanced Double-Strand Break Formation at Abasic Sites within Clustered Lesions in Nucleosome Core Particles.

    PubMed

    Banerjee, Samya; Chakraborty, Supratim; Jacinto, Marco Paolo; Paul, Michael D; Balster, Morgan V; Greenberg, Marc M

    2017-01-10

    DNA is rapidly cleaved under mild alkaline conditions at apyrimidinic/apurinic sites, but the half-life is several weeks in phosphate buffer (pH 7.5). However, abasic sites are ∼100-fold more reactive within nucleosome core particles (NCPs). Histone proteins catalyze the strand scission, and at superhelical location 1.5, the histone H4 tail is largely responsible for the accelerated cleavage. The rate constant for strand scission at an abasic site is enhanced further in a nucleosome core particle when it is part of a bistranded lesion containing a proximal strand break. Cleavage of this form results in a highly deleterious double-strand break. This acceleration is dependent upon the position of the abasic lesion in the NCP and its structure. The enhancement in cleavage rate at an apurinic/apyrimidinic site rapidly drops off as the distance between the strand break and abasic site increases and is negligible once the two forms of damage are separated by 7 bp. However, the enhancement of the rate of double-strand break formation increases when the size of the gap is increased from one to two nucleotides. In contrast, the cleavage rate enhancement at 2-deoxyribonolactone within bistranded lesions is more modest, and it is similar in free DNA and nucleosome core particles. We postulate that the enhanced rate of double-strand break formation at bistranded lesions containing apurinic/apyrimidinic sites within nucleosome core particles is a general phenomenon and is due to increased DNA flexibility.

  11. Remedial action plan and site design for stabilization of the inactive Uranium Mill Tailings Site at Lowman, Idaho

    SciTech Connect

    Not Available

    1991-09-01

    The DOE proposes to achieve compliance with the proposed EPA groundwater protection standards (Subparts A and B of 40 CFR 192) by meeting the EPA maximum concentration limits (MCLs) or background concentrations for designated hazardous constituents in groundwater in the uppermost aquifer (alluvium/weathered granodiorite) at the point of compliance (POC) at the Lowman disposal site near Lowman, Idaho. The proposed remedial action in conjunction with existing hydrogeological conditions at the Lowman site will ensure sufficient protection of human health and the environment. The DOE has concluded that the EPA groundwater protection standards will be met at the POC because, with the exception of antimony, none of the hazardous constituents that exceed laboratory method detection limits within the radioactive sand pore fluids were above the proposed concentration limits. The DOE has demonstrated that antimony will meet the proposed concentration limits at the POC through attenuation in subsoils beneath the disposal cell and by dilution in groundwater underflow. The Lowman processing site is in compliance with Subpart B of 40 CFR 192 because statistical analyses of groundwater samples indicate no groundwater contamination.

  12. Drift design methodology and preliminary application for the Yucca Mountain Site Characterization Project; Yucca Mountain Site Characterization Project

    SciTech Connect

    Hardy, M.P.; Bauer, S.J.

    1991-12-01

    Excavation stability in an underground nuclear waste repository is required during construction, emplacement, retrieval (if required), and closure phases to ensure worker health and safety, and to prevent development of potential pathways for radionuclide migration in the post-closure period. Stable excavations are developed by appropriate excavation procedures, design of the room shape, design and installation of rock support reinforcement systems, and implementation of appropriate monitoring and maintenance programs. In addition to the loads imposed by the in situ stress field, the repository drifts will be impacted by thermal loads developed after waste emplacement and, periodically, by seismic loads from naturally occurring earthquakes and underground nuclear events. A priori evaluation of stability is required for design of the ground support system, to confirm that the thermal loads are reasonable, and to support the license application process. In this report, a design methodology for assessing drift stability is presented. This is based on site conditions, together with empirical and analytical methods. Analytical numerical methods are emphasized at this time because empirical data are unavailable for excavations in welded tuff either at elevated temperatures or under seismic loads. The analytical methodology incorporates analysis of rock masses that are systematically jointed, randomly jointed, and sparsely jointed. In situ thermal and seismic loads are considered. Methods of evaluating the analytical results and estimating ground support requirements for all the full range of expected ground conditions are outlines. The results of a preliminary application of the methodology using the limited available data are presented. 26 figs., 55 tabs.

  13. Design Guidelines for Teaching about Design Guidelines for Educational WWW Sites.

    ERIC Educational Resources Information Center

    Collis, Betty; Winnips, Koos

    This paper describes how the authors use a mixture of World Wide Web (WWW)-based functionalities and new didactics to teach educational technology students at the University of Twente (Netherlands) about the design of WWW-based learning environments. Topics discussed include: (1) the content of courses about the design of educational WWW…

  14. Remedial action plan and site design for stabilization of the inactive uranium mill tailings site at Tuba City, Arizona. [Uranium Mill Tailings Remedial Action (UMTRA) Project

    SciTech Connect

    Not Available

    1987-05-01

    This appendix assesses the present conditions and data for the inactive uranium mill site near Tuba City, Arizona. It consolidates available engineering, radiological, geotechnical, hydrological, meterological, and other information pertinent to the design of the Remedial Action Plan (RAP). The data characterize conditions at the mill and tailings site so that the Remedial Action Contractor (RAC) may complete final designs of the remedial actions.

  15. Locked by Design: A Conformationally Constrained Transglutaminase Tag Enables Efficient Site-Specific Conjugation.

    PubMed

    Siegmund, Vanessa; Schmelz, Stefan; Dickgiesser, Stephan; Beck, Jan; Ebenig, Aileen; Fittler, Heiko; Frauendorf, Holm; Piater, Birgit; Betz, Ulrich A K; Avrutina, Olga; Scrima, Andrea; Fuchsbauer, Hans-Lothar; Kolmar, Harald

    2015-11-02

    Based on the crystal structure of a natural protein substrate for microbial transglutaminase, an enzyme that catalyzes protein crosslinking, a recognition motif for site-specific conjugation was rationally designed. Conformationally locked by an intramolecular disulfide bond, this structural mimic of a native conjugation site ensured efficient conjugation of a reporter cargo to the therapeutic monoclonal antibody cetuximab without erosion of its binding properties. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Aerosol measurements at the Southern Great Plains Site: Design and surface installation

    SciTech Connect

    Leifer, R.; Knuth, R.H.; Guggenheim, S.F.; Albert, B.

    1996-04-01

    To impropve the predictive capabilities of the Atmospheric Radiation Measurements (ARM) program radiation models, measurements of awserosol size distributions, condensation particle concentrations, aerosol scattering coefficients at a number of wavelenghts, and the aerosol absorption coefficients are needed at the Southern Great Plains (SGP) site. Alos, continuous measurements of ozone concnetrations are needed for model validation. The environmental Measuremenr Laboratory (EMK) has the responsibility to establish the surface aerosol measurements program at the SGP site. EML has designed a special sampling manifold.

  17. Are biophilic-designed site office buildings linked to health benefits and high performing occupants?

    PubMed

    Gray, Tonia; Birrell, Carol

    2014-11-26

    This paper discusses the first phase of a longitudinal study underway in Australia to ascertain the broad health benefits of specific types of biophilic design for workers in a building site office. A bespoke site design was formulated to include open plan workspace, natural lighting, ventilation, significant plants, prospect and views, recycled materials and use of non-synthetic materials. Initial data in the first three months was gathered from a series of demographic questions and from interviews and observations of site workers. Preliminary data indicates a strong positive effect from incorporating aspects of biophilic design to boost productivity, ameliorate stress, enhance well-being, foster a collaborative work environment and promote workplace satisfaction, thus contributing towards a high performance workspace. The longitudinal study spanning over two years will track human-plant interactions in a biophilic influenced space, whilst also assessing the concomitant cognitive, social, psychological and physical health benefits for workers.

  18. Design and control strategy for a hybrid green energy system for mobile telecommunication sites

    NASA Astrophysics Data System (ADS)

    Okundamiya, Michael S.; Emagbetere, Joy O.; Ogujor, Emmanuel A.

    2014-07-01

    The rising energy costs and carbon footprint of operating mobile telecommunication sites in the emerging world have increased research interests in green technology. The intermittent nature of most green energy sources creates the problem of designing the optimum configuration for a given location. This study presents the design analysis and control strategy for a cost effective and reliable operation of the hybrid green energy system (HGES) for GSM base transceiver station (BTS) sites in isolated regions. The design constrains the generation and distribution of power to reliably satisfy the energy demand while ensuring safe operation of the system. The overall process control applies the genetic algorithm-based technique for optimal techno-economic sizing of system's components. The process simulation utilized meteorological data for 3 locations (Abuja, Benin City and Sokoto) with varying climatic conditions in Nigeria. Simulation results presented for green GSM BTS sites are discussed and compared with existing approaches.

  19. Are Biophilic-Designed Site Office Buildings Linked to Health Benefits and High Performing Occupants?

    PubMed Central

    Gray, Tonia; Birrell, Carol

    2014-01-01

    This paper discusses the first phase of a longitudinal study underway in Australia to ascertain the broad health benefits of specific types of biophilic design for workers in a building site office. A bespoke site design was formulated to include open plan workspace, natural lighting, ventilation, significant plants, prospect and views, recycled materials and use of non-synthetic materials. Initial data in the first three months was gathered from a series of demographic questions and from interviews and observations of site workers. Preliminary data indicates a strong positive effect from incorporating aspects of biophilic design to boost productivity, ameliorate stress, enhance well-being, foster a collaborative work environment and promote workplace satisfaction, thus contributing towards a high performance workspace. The longitudinal study spanning over two years will track human-plant interactions in a biophilic influenced space, whilst also assessing the concomitant cognitive, social, psychological and physical health benefits for workers. PMID:25431874

  20. 41 CFR 102-76.20 - What issues must Federal agencies consider in providing site planning and landscape design services?

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... agencies consider in providing site planning and landscape design services? 102-76.20 Section 102-76.20...) FEDERAL MANAGEMENT REGULATION REAL PROPERTY 76-DESIGN AND CONSTRUCTION Design and Construction § 102-76.20 What issues must Federal agencies consider in providing site planning and landscape design services? In...

  1. System design package for solar heating and cooling site data acquisition subsystem

    NASA Technical Reports Server (NTRS)

    1978-01-01

    The Site Data Acquisition Subsystem (SDAS) designed to collect data from sensors located on residential or commercial buildings using a solar heating and/or cooling system is described. It takes the data obtained from sensors located on the solar system, processes the data into suitable format, stores the data for a period of time, and provides the capability for either telephone retrieval by the central data processing system or manual retrieval of the data for transfer to a central site. The SDAS is also designed so that it will not degrade the operation of the solar heating/cooling system which it is monitoring.

  2. Subsystem design package for Mod 2 site data acquisition system: Solar heating and cooling

    NASA Technical Reports Server (NTRS)

    1977-01-01

    The Mod II Site Data Acquisition Subsystem (SDAS) is designed to collect data from sensors located on residential or commercial buildings using a solar heating and/or cooling system. The SDAS takes the data obtained from sensors located on the solar heating and/or cooling system, processes the data into a suitable format, stores the data for a period of time, and provides the capability for both telephone retrieval by the Central Data Processing System (CDPS) and manual retrieval of the data for transfer to the central site. The unit is designed so it will not degrade the operation of the solar heating/cooling system which it is monitoring.

  3. Compounds designed to fit a site of known structure in human haemoglobin.

    PubMed Central

    Beddell, C R; Goodford, P J; Norrington, F E; Wilkinson, S; Wootton, R

    1976-01-01

    1 The three-dimensional coordinates of the atoms in human haemoglobin are known, and there is a specific site in the deoxygenated form of the protein at which 2,3-diphosphoglycerate (DPG) interacts. 2 Molecular models of this site have been constructed and used to design compounds which should bind to the deoxy conformation and stabilize it. These compounds should therby promote oxygen liberation, as does DPG. 3The compounds so designed were found to promote oxygen liberation. Their relative potencies, as assessed by sigmoidal dose-response curves, are in the predicted sequence. PMID:938794

  4. Two interacting binding sites for quinacrine derivatives in the active site of trypanothione reductase – a template for drug design

    PubMed Central

    Saravanamuthu, Ahilan; Vickers, Tim J.; Bond, Charles S.; Peterson, Mark R.; Hunter, William N.; Fairlamb, Alan H.

    2012-01-01

    SUMMARY Trypanothione reductase is a key enzyme in the trypanothione-based redox metabolism of pathogenic trypanosomes. Since this system is absent in humans, being replaced with glutathione and glutathione reductase, it offers a target for selective inhibition. The rational design of potent inhibitors requires accurate structures of enzyme-inhibitor complexes, but this is lacking for trypanothione reductase. We therefore used quinacrine mustard, an alkylating derivative of the competitive inhibitor quinacrine, to probe the active site of this dimeric flavoprotein. Quinacrine mustard irreversibly inactivates Trypanosoma cruzi trypanothione reductase, but not human glutathione reductase, in a time-dependent manner with a stoichiometry of two inhibitors bound per monomer. The rate of inactivation is dependent upon the oxidation state of trypanothione reductase, with the NADPH-reduced form being inactivated significantly faster than the oxidised form. Inactivation is slowed by clomipramine and a melarsen oxide-trypanothione adduct (both are competitive inhibitors) but accelerated by quinacrine. The structure of the trypanothione reductase-quinacrine mustard adduct was determined to 2.7 Å, revealing two molecules of inhibitor bound in the trypanothione-binding site. The acridine moieties interact with each other through π-stacking effects, and one acridine interacts in a similar fashion with a tryptophan residue. These interactions provide a molecular explanation for the differing effects of clomipramine and quinacrine on inactivation by quinacrine mustard. Synergism with quinacrine occurs as a result of these planar acridines being able to stack together in the active site cleft, thereby gaining an increased number of binding interactions, whereas antagonism occurs with non-planar molecules, such as clomipramine, where stacking is not possible. PMID:15102853

  5. Design and implementation of an identification system in construction site safety for proactive accident prevention.

    PubMed

    Yang, Huanjia; Chew, David A S; Wu, Weiwei; Zhou, Zhipeng; Li, Qiming

    2012-09-01

    Identifying accident precursors using real-time identity information has great potential to improve safety performance in construction industry, which is still suffering from day to day records of accident fatality and injury. Based on the requirements analysis for identifying precursor and the discussion of enabling technology solutions for acquiring and sharing real-time automatic identification information on construction site, this paper proposes an identification system design for proactive accident prevention to improve construction site safety. Firstly, a case study is conducted to analyze the automatic identification requirements for identifying accident precursors in construction site. Results show that it mainly consists of three aspects, namely access control, training and inspection information and operation authority. The system is then designed to fulfill these requirements based on ZigBee enabled wireless sensor network (WSN), radio frequency identification (RFID) technology and an integrated ZigBee RFID sensor network structure. At the same time, an information database is also designed and implemented, which includes 15 tables, 54 queries and several reports and forms. In the end, a demonstration system based on the proposed system design is developed as a proof of concept prototype. The contributions of this study include the requirement analysis and technical design of a real-time identity information tracking solution for proactive accident prevention on construction sites. The technical solution proposed in this paper has a significant importance in improving safety performance on construction sites. Moreover, this study can serve as a reference design for future system integrations where more functions, such as environment monitoring and location tracking, can be added. Copyright © 2011 Elsevier Ltd. All rights reserved.

  6. Conceptual design for an on-site spent-fuel transfer system

    SciTech Connect

    Pennington, C.W.; Neider, T.J.; Guerra, G.V.; Margotta, K.V. )

    1991-09-01

    Transnuclear, Inc. has developed two conceptual designs for transferring spent fuel from a nuclear plant's fuel storage pool to large storage casks by means of a small transfer cask. The transfer of spent fuel would be performed external to the nuclear plant, rather than within the plant's fuel building or transport bay. The systems can also be used to transfer fuel between storage casks and transport casks. Both systems meet the design considerations for on-site spent-fuel transfer systems outlined in EPRI Report NP-6425, Design Considerations for On-Site Spent-Fuel Transfer Systems.'' One of the designs involves a dry transfer into the storage cask, while the other uses a wet transfer method. Both systems have been evaluated for economic and technical feasibility, licensability and practicality. This report describes each of the conceptual designs and outlines the operating procedures for each system. In addition, a risk and accident assessment, a logistical evaluation, and an economic analysis including target cost evaluations and system cost estimates are included. This report shows that it is technically feasible and that costs are economically justifiable to perform on-site spent-fuel transfers using small transfer casks. However, the final design selection for the transfer system could depend on the individual utility's operational preferences and the unique facility features that already exist at each nuclear plant. 9 tabs.

  7. Guide to radiological accident considerations for siting and design of DOE nonreactor nuclear facilities

    SciTech Connect

    Elder, J.; Graf, J.M.

    1984-01-01

    DOE Office of Nuclear Safety has sponsored preparation of a guidance document to aid field offices and contractors in their analyses of consequences of postulated major accidents. The guide addresses the requirements of DOE Orders 5480.1A, Chapter V, and 6430.1, including the general requirement that DOE nuclear facilities be sited, designed, and operated in accordance with standards, codes, and guides consistent with those applied to comparable licensed nuclear facilities. The guide includes both philosophical and technical information in the areas of: siting guidelines doses applied to an offsite reference person; consideration also given to an onsite reference person; physical parameters, models, and assumptions to be applied when calculating doses for comparison to siting criteria; and potential accident consequences other than radiological dose to a reference person which might affect siting and major design features of the facility, such as environmental contamination, population dose, and associated public health effects. Recommendations and/or clarifications are provided where this could be done without adding new requirements. In this regard, the guide is considered a valuable aid to the safety analyst, especially where requirements have been subject to inconsistent interpretation or where analysis methods are in transition, such as use of dose model (ICRP 2 or ICRP 30) or use of probabilistic methods of risk analysis in the siting and design of nuclear facilities.

  8. HTP-OligoDesigner: An Online Primer Design Tool for High-Throughput Gene Cloning and Site-Directed Mutagenesis.

    PubMed

    Camilo, Cesar M; Lima, Gustavo M A; Maluf, Fernando V; Guido, Rafael V C; Polikarpov, Igor

    2016-01-01

    Following burgeoning genomic and transcriptomic sequencing data, biochemical and molecular biology groups worldwide are implementing high-throughput cloning and mutagenesis facilities in order to obtain a large number of soluble proteins for structural and functional characterization. Since manual primer design can be a time-consuming and error-generating step, particularly when working with hundreds of targets, the automation of primer design process becomes highly desirable. HTP-OligoDesigner was created to provide the scientific community with a simple and intuitive online primer design tool for both laboratory-scale and high-throughput projects of sequence-independent gene cloning and site-directed mutagenesis and a Tm calculator for quick queries.

  9. Role of the Nfo and ExoA apurinic/apyrimidinic endonucleases in radiation resistance and radiation-induced mutagenesis of Bacillus subtilis spores.

    PubMed

    Moeller, Ralf; Setlow, Peter; Pedraza-Reyes, Mario; Okayasu, Ryuichi; Reitz, Günther; Nicholson, Wayne L

    2011-06-01

    The roles of DNA repair by apurinic/apyrimidinic (AP) endonucleases alone, and together with DNA protection by α/β-type small acid-soluble spore proteins (SASP), in Bacillus subtilis spore resistance to different types of radiation have been studied. Spores lacking both AP endonucleases (Nfo and ExoA) and major SASP were significantly more sensitive to 254-nm UV-C, environmental UV (>280 nm), X-ray exposure, and high-energy charged (HZE)-particle bombardment and had elevated mutation frequencies compared to those of wild-type spores and spores lacking only one or both AP endonucleases or major SASP. These findings further implicate AP endonucleases and α/β-type SASP in repair and protection, respectively, of spore DNA against effects of UV and ionizing radiation.

  10. Site layout and balance of plant design for an accelerator-driven materials processing complex

    SciTech Connect

    Cunliffe, J.; Taussig, R.; Ghose, S.

    1995-10-01

    High energy proton beam accelerators are under consideration for use in radioisotope production, surplus weapons material destruction, radioactive waste transmutation, and thorium-based energy conversion cycles. While there are unique aspects to each of these applications that must be accommodated in the design of the associated facility, all share a set of fundamental characteristics that in large measure dictate the site layout features and many balance-of-plant (BOP) design requirements found to be common to all. This paper defines these key design determinants and goes on to discuss the manner in which they have been accommodated in the pre-conceptual design for a particular materials production application. An estimate of the costs associated with this BOP design is also presented with the aim of guiding future evaluations where the basic plant designs are similar to that of this specific case.

  11. Design, synthesis, and characterization of nucleosomes containing site-specific DNA damage.

    PubMed

    Taylor, John-Stephen

    2015-12-01

    How DNA damaged is formed, recognized, and repaired in chromatin is an area of intense study. To better understand the structure activity relationships of damaged chromatin, mono and dinucleosomes containing site-specific damage have been prepared and studied. This review will focus on the design, synthesis, and characterization of model systems of damaged chromatin for structural, physical, and enzymatic studies.

  12. Understanding and Designing for Interactional Privacy Needs within Social Networking Sites

    ERIC Educational Resources Information Center

    Wisniewski, Pamela J.

    2012-01-01

    "Interpersonal boundary regulation" is a way to optimize social interactions when sharing and connecting through Social Networking Sites (SNSs). The theoretical foundation of much of my research comes from Altman's work on privacy management in the physical world. Altman believed that "we should attempt to design responsive…

  13. 10 CFR 435.6 - Sustainable principles for siting, design and construction. [Reserved

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 10 Energy 3 2013-01-01 2013-01-01 false Sustainable principles for siting, design and construction. 435.6 Section 435.6 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY STANDARDS FOR NEW FEDERAL LOW-RISE RESIDENTIAL BUILDINGS Mandatory Energy Efficiency Standards for Federal Low-Rise...

  14. 10 CFR 435.6 - Sustainable principles for siting, design and construction. [Reserved

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 10 Energy 3 2011-01-01 2011-01-01 false Sustainable principles for siting, design and construction. 435.6 Section 435.6 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY STANDARDS FOR NEW FEDERAL LOW-RISE RESIDENTIAL BUILDINGS Mandatory Energy Efficiency Standards for Federal Low-Rise...

  15. 10 CFR 433.6 - Sustainable principles for siting, design and construction. [Reserved

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 10 Energy 3 2014-01-01 2014-01-01 false Sustainable principles for siting, design and construction. 433.6 Section 433.6 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY STANDARDS FOR NEW FEDERAL COMMERCIAL AND MULTI-FAMILY HIGH-RISE RESIDENTIAL BUILDINGS § 433.6 Sustainable principles...

  16. 10 CFR 435.6 - Sustainable principles for siting, design and construction. [Reserved

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 10 Energy 3 2010-01-01 2010-01-01 false Sustainable principles for siting, design and construction. 435.6 Section 435.6 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY STANDARDS FOR NEW FEDERAL LOW-RISE RESIDENTIAL BUILDINGS Mandatory Energy Efficiency Standards for Federal Low-Rise...

  17. 10 CFR 433.6 - Sustainable principles for siting, design and construction. [Reserved

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 10 Energy 3 2013-01-01 2013-01-01 false Sustainable principles for siting, design and construction. 433.6 Section 433.6 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY STANDARDS FOR NEW FEDERAL COMMERCIAL AND MULTI-FAMILY HIGH-RISE RESIDENTIAL BUILDINGS § 433.6 Sustainable principles...

  18. 10 CFR 435.6 - Sustainable principles for siting, design and construction. [Reserved

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 10 Energy 3 2012-01-01 2012-01-01 false Sustainable principles for siting, design and construction. 435.6 Section 435.6 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY STANDARDS FOR NEW FEDERAL LOW-RISE RESIDENTIAL BUILDINGS Mandatory Energy Efficiency Standards for Federal Low-Rise...

  19. 10 CFR 435.6 - Sustainable principles for siting, design and construction. [Reserved

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 10 Energy 3 2014-01-01 2014-01-01 false Sustainable principles for siting, design and construction. 435.6 Section 435.6 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY STANDARDS FOR NEW FEDERAL LOW-RISE RESIDENTIAL BUILDINGS Mandatory Energy Efficiency Standards for Federal Low-Rise...

  20. 10 CFR 433.6 - Sustainable principles for siting, design and construction. [Reserved

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 10 Energy 3 2012-01-01 2012-01-01 false Sustainable principles for siting, design and construction. 433.6 Section 433.6 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY STANDARDS FOR NEW FEDERAL COMMERCIAL AND MULTI-FAMILY HIGH-RISE RESIDENTIAL BUILDINGS § 433.6 Sustainable principles...

  1. Conceptual design report, TWRS Privatization Phase I, site development and roads, subproject W-505

    SciTech Connect

    Singh, G.

    1997-06-05

    This document includes Conceptual Design Report (CDR) for the site development, construction of new roads and improvements at existing road intersections, habitat mitigation, roadway lighting, and construction power needed for the construction of two Private Contractor (PC) Facilities. Approximately 50 hectare (124 acres) land parcel, east of the Grout Facility, is planned for the PC facilities.

  2. Understanding and Designing for Interactional Privacy Needs within Social Networking Sites

    ERIC Educational Resources Information Center

    Wisniewski, Pamela J.

    2012-01-01

    "Interpersonal boundary regulation" is a way to optimize social interactions when sharing and connecting through Social Networking Sites (SNSs). The theoretical foundation of much of my research comes from Altman's work on privacy management in the physical world. Altman believed that "we should attempt to design responsive…

  3. 75 FR 54497 - Ocean Dumping; Guam Ocean Dredged Material Disposal Site Designation

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-08

    ... at the site will be limited to a maximum of 1 million cubic yards (764,555 cubic meters) per calendar...). However, all dredged material must be discharged within a smaller 3,280 foot (1,000 meter) diameter...,790 feet (2,680 meters). D. Disposal Volume Limit G-DODS is designated for a maximum annual dredged...

  4. Geohydrologic aspects for siting and design of low-level radioactive-waste disposal

    USGS Publications Warehouse

    Bedinger, M.S.

    1989-01-01

    The objective for siting and design of low-level radioactive-waste repository sites is to isolate the waste from the biosphere until the waste no longer poses an unacceptable hazard as a result of radioactive decay. Low-level radioactive waste commonly is isolated at shallow depths with various engineered features to stabilize the waste and to reduce its dissolution and transport by ground water. The unsaturated zone generally is preferred for isolating the waste. Low-level radioactive waste may need to be isolated for 300 to 500 years. Maintenance and monitoring of the repository site are required by Federal regulations for only the first 100 years. Therefore, geohydrology of the repository site needs to provide natural isolation of the waste for the hazardous period following maintenance of the site. Engineering design of the repository needs to be compatible with the natural geohydrologic conditions at the site. Studies at existing commercial and Federal waste-disposal sites provide information on the problems encountered and the basis for establishing siting guidelines for improved isolation of radioactive waste, engineering design of repository structures, and surveillance needs to assess the effectiveness of the repositories and to provide early warning of problems that may require remedial action. Climate directly affects the hydrology of a site and probably is the most important single factor that affects the suitability of a site for shallow-land burial of low-level radioactive waste. Humid and subhumid regions are not well suited for shallow isolation of low-level radioactive waste in the unsaturated zone; arid regions with zero to small infiltration from precipitation, great depths to the water table, and long flow paths to natural discharge areas are naturally well suited to isolation of the waste. The unsaturated zone is preferred for isolation of low-level radioactive waste. The guiding rationale is to minimize contact of water with the waste and to

  5. GMT site, enclosure, and facilities design and development overview and update

    NASA Astrophysics Data System (ADS)

    Teran, Jose; Burgett, William S.; Grigel, Eric; Bigelow, Bruce C.; Donoso, Eduardo; Figueroa, Francisco

    2016-07-01

    The Giant Magellan Telescope (GMT) will have a 25.4-meter diameter effective aperture, and is one the three currently planned next generation extremely large telescopes (ELTs). The GMT will be located at the summit of Cerro Campanas at the Las Campanas Observatory (LCO) in Chile, one the world's best observing sites. This paper provides an overview of the site master plan comprising site infrastructure, enclosure, and facilities, and outlines the analysis of alternative trade studies that will lead to the final design. Also presented is an update of the site infrastructure development and preconstruction activities currently underway that will be completed prior to the beginning of enclosure construction near the end of 2016.

  6. Remedial action plan and site conceptual design for stabilization of the inactive uranium mill tailings sites at Rifle, Colorado. Appendix D, Final report

    SciTech Connect

    1992-02-01

    This appendix assesses the present conditions and data gathered about the two designated inactive uranium mill tailings sites near Rifle, Colorado, and the proposed disposal site six miles north of Rifle in the area of Estes Gulch. It consolidates available engineering, radiological, geotechnical, hydrological, meteorological, and other information pertinent to the design of the Remedial Action Plan (RAP). The data characterize conditions at the mill, tailings, and disposal site so that the Remedial Action Contractor (RAC) may complete final designs for the remedial actions.

  7. Conceptual Design Report: Nevada Test Site Mixed Waste Disposal Facility Project

    SciTech Connect

    NSTec Environmental Management

    2009-01-31

    Environmental cleanup of contaminated nuclear weapons manufacturing and test sites generates radioactive waste that must be disposed. Site cleanup activities throughout the U.S. Department of Energy (DOE) complex are projected to continue through 2050. Some of this waste is mixed waste (MW), containing both hazardous and radioactive components. In addition, there is a need for MW disposal from other mission activities. The Waste Management Programmatic Environmental Impact Statement Record of Decision designates the Nevada Test Site (NTS) as a regional MW disposal site. The NTS has a facility that is permitted to dispose of onsite- and offsite-generated MW until November 30, 2010. There is not a DOE waste management facility that is currently permitted to dispose of offsite-generated MW after 2010, jeopardizing the DOE environmental cleanup mission and other MW-generating mission-related activities. A mission needs document (CD-0) has been prepared for a newly permitted MW disposal facility at the NTS that would provide the needed capability to support DOE's environmental cleanup mission and other MW-generating mission-related activities. This report presents a conceptual engineering design for a MW facility that is fully compliant with Resource Conservation and Recovery Act (RCRA) and DOE O 435.1, 'Radioactive Waste Management'. The facility, which will be located within the Area 5 Radioactive Waste Management Site (RWMS) at the NTS, will provide an approximately 20,000-cubic yard waste disposal capacity. The facility will be licensed by the Nevada Division of Environmental Protection (NDEP).

  8. Custom-Designed Molecular Scissors for Site-Specific Manipulation of the Plant and Mammalian Genomes

    NASA Astrophysics Data System (ADS)

    Kandavelou, Karthikeyan; Chandrasegaran, Srinivasan

    Zinc finger nucleases (ZFNs) are custom-designed molecular scissors, engineered to cut at specific DNA sequences. ZFNs combine the zinc finger proteins (ZFPs) with the nonspecific cleavage domain of the FokI restriction enzyme. The DNA-binding specificity of ZFNs can be easily altered experimentally. This easy manipulation of the ZFN recognition specificity enables one to deliver a targeted double-strand break (DSB) to a genome. The targeted DSB stimulates local gene targeting by several orders of magnitude at that specific cut site via homologous recombination (HR). Thus, ZFNs have become an important experimental tool to make site-specific and permanent alterations to genomes of not only plants and mammals but also of many other organisms. Engineering of custom ZFNs involves many steps. The first step is to identify a ZFN site at or near the chosen chromosomal target within the genome to which ZFNs will bind and cut. The second step is to design and/or select various ZFP combinations that will bind to the chosen target site with high specificity and affinity. The DNA coding sequence for the designed ZFPs are then assembled by polymerase chain reaction (PCR) using oligonucleotides. The third step is to fuse the ZFP constructs to the FokI cleavage domain. The ZFNs are then expressed as proteins by using the rabbit reticulocyte in vitro transcription/translation system and the protein products assayed for their DNA cleavage specificity.

  9. Designing a web site for high school geoscience teaching in Iceland

    NASA Astrophysics Data System (ADS)

    Douglas, George R.

    1998-08-01

    The need to construct an earth science teaching site on the web prompted a survey of existing sites which, in spite of containing much of value, revealed many weaknesses in basic design, particularly as regards the organisation of links to information resources. Few web sites take into consideration the particular pedagogic needs of the high school science student and there has, as yet, been little serious attempt to exploit and organise the more outstanding advantages offered by the internet to science teaching, such as accessing real-time data. A web site has been constructed which, through basic design, enables students to access relevant information resources over a wide range of subjects and topics easily and rapidly, while at the same time performing an instructional role in how to handle both on-line and off-line resources. Key elements in the design are selection and monitoring by the teacher, task oriented pages and the use of the Dewey decimal classification system. The intention is to increase gradually the extent to which most teaching tasks are carried out via the web pages, in the belief that they can become an efficient central point for all the earth science curriculum.

  10. Remedial action and site design for stabilization of the inactive uranium mill tailings sites at Slick Rock, Colorado. Attachment 2, Geology report: Appendix B, Preliminary final

    SciTech Connect

    Not Available

    1994-03-01

    Detailed investigations of geologic, geomorphic, and seismic conditions at the Burro Canyon site were conducted by the US Department of Energy (DOE) as a disposal site for the tailings at two processing sites near the Slick Rock, Colorado, post office. The purposes of these studies are basic site characterization and identification of potential geologic hazards that could affect long-term site stability. Subsequent engineering studies (e.g., analyses of hydrologic and liquefaction hazards) used the data developed in these studies. The geomorphic analysis was employed in the design of effective erosion protection. Studies of the regional and local seismotectonic setting, which included a detailed search for possible capable faults within a 65-km radius of the site, provided the basis for seismic design parameters.

  11. Compressed-air energy storage preliminary design and site development program in an aquifer. Volume 5, Part 1: Turbomachinery design

    NASA Astrophysics Data System (ADS)

    Berman, P. A.; Bonk, J. S.; Kobett, W. F.; Kosanovich, N. S.; Long, L. J.; Marinacci, D. J.

    1982-11-01

    The development of the design approach for a combustion turbine heat cycle and the major mechanical equipment for use by an electric utility at a selected aquifer air storage site is documented. A compressed air energy storage (CAES) system utilizes off peak electric power, available from base load power plants, to store compressed air underground in an aquifer. During subsequent periods, the stored air is extracted from the aquifer and used as an air supply for a generating combustion turbine expander. The aquifer has an initial discovery pressure of 840 psia. An initial air injection temperature of 1500 F was selected. The major mechanical equipment considered includes: the turbine motor/generator compressor train, intercooler and aftercooler system, and the exhaust gas regenerator. The cycle and machinery configuration and the specific mechanical equipment were selected for their Media site characteristics. These characteristics and the effect of component interdependency are considered when a conservative component design approach is established which satisfies the Media site CAES system requirements.

  12. Event-based design tool for construction site erosion and sediment controls

    NASA Astrophysics Data System (ADS)

    Trenouth, William R.; Gharabaghi, Bahram

    2015-09-01

    This paper provides additional discussion surrounding the novel event-based soil loss models developed by Trenouth and Gharabaghi (2015) for the design of erosion and sediment controls (ESCs) for various phases of construction - from pre-development to post-development conditions. The datasets for the study were obtained from three Ontario sites - Greensborough, Cookstown, and Alcona - in addition to datasets mined from the literature for three additional sites - Treynor, Iowa, Coshocton, Ohio and Cordoba, Spain. Model performances were evaluated for each of the study sites, and quantified using commonly-reported statistics. This work is nested within a broader conceptual framework, which includes the estimation of ambient receiving water quality, the prediction of event mean runoff quality for a given design storm, and the calculation of the required level of protection using adequate ESCs to meet receiving water quality guidelines. These models allow design engineers and regulatory agencies to assess the potential risk of ecological damage to receiving waters due to inadequate soil erosion and sediment control practices using dynamic scenario forecasting when considering rapidly changing land use conditions during various phases of construction, typically for a 2- or 5-year design storm return period.

  13. Remedial Action Plan and site design for stabilization of the inactive uranium mill tailings site at Mexican Hat, Utah: Appendix D. Final report

    SciTech Connect

    1988-07-01

    This appendix is an assessment of the present conditions of the inactive uranium mill site near Mexican Hat, Utah. It consolidates available engineering, radiological, geotechnical, hydrological, meteorological, and other information pertinent to the design of the Remedial Action Plan. Plan is to characterize the conditions at the mill and tailings site so that the Remedial Action Contractor may complete final designs of the remedial action.

  14. Remedial action plan and site design for stabilization of the inactive uranium mill tailings sites at Slick Rock, Colorado. Remedial action selection report, Appendix B

    SciTech Connect

    Not Available

    1993-07-01

    The Slick Rock uranium mill tailings sites are located near the small town of Slick Rock, in San Miguel County, Colorado. There are two designated UMTRA sites at Slick Rock, the Union Carbide (UC) site and the North Continent (NC) site. Both sites are adjacent to the Dolores River. The UC site is approximately 1 mile (mi) [2 kilometers (km)] downstream of the NC site. Contaminated materials cover an estimated 55 acres (ac) [22 hectares (ha)] at the UC site and 12 ac (4.9 ha) at the NC site. The sites contain former mill building concrete foundations, tailings piles, demolition debris, and areas contaminated by windblown and waterborne radioactive materials. The total estimated volume of contaminated materials is approximately 620, 000 cubic yards (yd{sup 3}) [470,000 cubic meters (m{sup 3})]. In addition to the contamination at the two processing site areas, four vicinity properties were contaminated. Contamination associated with the UC and NC sites has leached into groundwater.

  15. Guide to radiological accident considerations for siting and design of DOE nonreactor nuclear facilities

    SciTech Connect

    Elder, J.C.; Graf, J.M.; Dewart, J.M.; Buhl, T.E.; Wenzel, W.J.; Walker, L.J.; Stoker, A.K.

    1986-01-01

    This guide was prepared to provide the experienced safety analyst with accident analysis guidance in greater detail than is possible in Department of Energy (DOE) Orders. The guide addresses analysis of postulated serious accidents considered in the siting and selection of major design features of DOE nuclear facilities. Its scope has been limited to radiological accidents at nonreactor nuclear facilities. The analysis steps addressed in the guide lead to evaluation of radiological dose to exposed persons for comparison with siting guideline doses. Other possible consequences considered are environmental contamination, population dose, and public health effects. Choices of models and parameters leading to estimation of source terms, release fractions, reduction and removal factors, dispersion and dose factors are discussed. Although requirements for risk analysis have not been established, risk estimates are finding increased use in siting of major nuclear facilities, and are discussed in the guide. 3 figs., 9 tabs.

  16. Region and site specific design guidelines for the northeastern Venezuela development

    SciTech Connect

    Gajardo, E.; Paga, M.; Sully, J.P.

    1996-12-31

    Several site specific studies were performed for the northeastern Venezuelan offshore development in order to determine the environmental parameters for the engineering design. The geotechnical parameters were obtained from the analysis of the soil borings, in situ tests and the high resolution seismic surveys. An evaluation of the seismic hazard was done, based on a detailed neotectonic study, the compilation of historical and instrumental seismological information, and an attenuation relationship developed for the studied area. Seismic response analyses were performed using the seismic parameters and dynamic characteristics obtained form the soil borings and in situ tests of shear waves. The met-ocean design loads of the local conditions were obtained from mathematical modeling based on in situ and regional measurements. This study defines the platforms design conditions based on a probabilistic reliability approach and it has been used, among others, as a basis for the earthquake design guidelines proposed by the International Standards Organization. A sensitivity analysis was performed starting from the uncertainties of all the input parameters, in order to obtain the best estimate average design spectrum and associated coefficient of variability. The specific approach was to maintain the same safety index adopted by API RP2A code, with a bias modification factor that relates the site nominal spectra with the spectral acceleration that would be used in order to satisfy the API requirements. The results of this approach make possible the use of site specific detailed environmental data and the proven standard procedures established on the API RP2 codes, with significant cost reduction in terms of both platform and foundation design. The considerable amount of tectonic, seismological, and geotechnical information was the principal factor allowing the above methodology.

  17. Design for hydrogen storage materials via observation of adsorption sites by computer tomography.

    PubMed

    Zhang, Li; Wang, Qi; Liu, Ying-Chun

    2007-05-03

    An effective method denoted as "computer tomography for materials" (mCT) was employed to study the adsorption sites inside metal-organic frameworks (MOFs) at any positions and any view angles. For MOF-5, the first adsorption site alpha(-COO)3 was clearly observed from the mCT images; it locates at the position where three -COO groups joined like a cup. There are four alpha(-COO)3 sites around the Zn4O cluster. Two of them located at the diagonal of the Zn4O cluster are in the same plane "A", whereas the other two equivalent adsorption sites are in another plane "B", which is about 5.4 A away from the plane A. It was found that the electronegativity of oxygen atoms is very important to the adsorption of hydrogen molecules. The hydrogen amount adsorbed in MOFs might be enhanced by introducing some strong electronegative atoms to the organic linkers or frameworks. On the basis of this point of view, five new MOF materials were designed. The adsorbed amounts both in number of hydrogen molecules per unit cell and weight uptake for all of the designed MOFs were calculated. The adsorption amounts of designed MOFs were improved, and the amount for MOF-d5 at 1 bar is as high as 3.7 wt %. It is nearly 5-6 times of that of MOF-5 as a whole. It can be observed that extra adsorption sites were formed in the pores and the effective occupation rate of pore space was obviously improved viewing from the mCT images. These results may give helpful suggestions for the synthetic experimentalists.

  18. Design Status and Applications of Small Reactors Without On-Site Refuelling

    SciTech Connect

    Kuznetsov, Vladimir

    2006-07-01

    Small reactors without on-site refuelling (SRWORs) are the reactors that can operate without reloading and shuffling of fuel for a reasonably long period with no refuelling equipment being present in the reactor and no fuel being stored at the site during reactor operation. By virtue of being small, transportable and requiring no operations with fuel from a customer, such reactors form an attractive domain for fuel or even NPP leasing. SRWORs could simplify the implementation of safeguards and provide certain guarantees of sovereignty to those countries that would agree to forego the development of the indigenous fuel cycle. About 30 concepts of such reactors are being analyzed or developed in 6 IAEA member states. Based on intermediate results of IAEA activities in support of the design and technology development for such reactors, the paper provides technical details on the design status, fuel cycle options and possible applications of SRWORs. (authors)

  19. Automated Design of Synthetic Ribosome Binding Sites to Precisely Control Protein Expression

    PubMed Central

    Salis, Howard M.; Mirsky, Ethan A.; Voigt, Christopher A.

    2009-01-01

    Microbial engineering often requires fine control over protein expression; for example, to connect genetic circuits 1-7 or control flux through a metabolic pathway 8-13. We have developed a predictive design method for synthetic ribosome binding sites that enables the rational control of a protein's production rate on a proportional scale. Experimental validation of over 100 predictions in Escherichia coli shows that the method is accurate to within a factor of 2.3 over a range of 100,000-fold. The design method also correctly predicts that reusing a ribosome binding site sequence in different genetic contexts can result in different protein expression levels. We demonstrate the method's utility by rationally optimizing a protein's expression level to connect a genetic sensor to a synthetic circuit. The proposed forward engineering approach will accelerate the construction and systematic optimization of large genetic systems. PMID:19801975

  20. The design of a Phase I non site-specific Centralized Interim Storage Facility

    SciTech Connect

    Stringer, J.; Kane, D.

    1997-10-28

    The Department of Energy (DOE), Office of Civilian Radioactive Waste Management (OCRWM) recently completed a Topical Safety Analysis Report (TSAR) for a Phase 1 non site specific Centralized Interim Storage Facility (CISF). The TSAR will be used in licensing the CISF when and if a site is designated. The combined Phase 1 and Phase 2 CISF will provide federal storage capability for 40,000 metric tons of uranium (MTU) Spent Nuclear Fuel (SNF) under the oversight of the DOE. The Phase 1 TSAR was submitted to the NRC on May 1, 1997 and is currently under review having been docketed on June 10, 1997. This paper generally describes the Phase 1 CISF design and its operations as presented in the CISF TSAR.

  1. Evolution of a designed retro-aldolase leads to complete active site remodeling.

    PubMed

    Giger, Lars; Caner, Sami; Obexer, Richard; Kast, Peter; Baker, David; Ban, Nenad; Hilvert, Donald

    2013-08-01

    Evolutionary advances are often fueled by unanticipated innovation. Directed evolution of a computationally designed enzyme suggests that pronounced molecular changes can also drive the optimization of primitive protein active sites. The specific activity of an artificial retro-aldolase was boosted >4,400-fold by random mutagenesis and screening, affording catalytic efficiencies approaching those of natural enzymes. However, structural and mechanistic studies reveal that the engineered catalytic apparatus, consisting of a reactive lysine and an ordered water molecule, was unexpectedly abandoned in favor of a new lysine residue in a substrate-binding pocket created during the optimization process. Structures of the initial in silico design, a mechanistically promiscuous intermediate and one of the most evolved variants highlight the importance of loop mobility and supporting functional groups in the emergence of the new catalytic center. Such internal competition between alternative reactive sites may have characterized the early evolution of many natural enzymes.

  2. Molecular dynamics explorations of active site structure in designed and evolved enzymes.

    PubMed

    Osuna, Sílvia; Jiménez-Osés, Gonzalo; Noey, Elizabeth L; Houk, K N

    2015-04-21

    This Account describes the use of molecular dynamics (MD) simulations to reveal how mutations alter the structure and organization of enzyme active sites. As proposed by Pauling about 70 years ago and elaborated by many others since then, biocatalysis is efficient when functional groups in the active site of an enzyme are in optimal positions for transition state stabilization. Changes in mechanism and covalent interactions are often critical parts of enzyme catalysis. We describe our explorations of the dynamical preorganization of active sites using MD, studying the fluctuations between active and inactive conformations normally concealed to static crystallography. MD shows how the various arrangements of active site residues influence the free energy of the transition state and relates the populations of the catalytic conformational ensemble to the enzyme activity. This Account is organized around three case studies from our laboratory. We first describe the importance of dynamics in evaluating a series of computationally designed and experimentally evolved enzymes for the Kemp elimination, a popular subject in the enzyme design field. We find that the dynamics of the active site is influenced not only by the original sequence design and subsequent mutations but also by the nature of the ligand present in the active site. In the second example, we show how microsecond MD has been used to uncover the role of remote mutations in the active site dynamics and catalysis of a transesterase, LovD. This enzyme was evolved by Tang at UCLA and Codexis, Inc., and is a useful commercial catalyst for the production of the drug simvastatin. X-ray analysis of inactive and active mutants did not reveal differences in the active sites, but relatively long time scale MD in solution showed that the active site of the wild-type enzyme preorganizes only upon binding of the acyl carrier protein (ACP) that delivers the natural acyl group to the active site. In the absence of bound ACP

  3. ONWI (Office of Nuclear Waste Isolation) 30% design review findings report for Exploratory Shaft Facility, Deaf Smith site

    SciTech Connect

    Not Available

    1987-05-08

    This document describes a review of the standards for the design of the high-level radioactive waste facility at the Deaf Smith, Texas site. It includes public comments and the official responses to the designs produced to date. (TEM)

  4. A sensitivity analysis of hazardous waste disposal site climatic and soil design parameters using HELP3

    SciTech Connect

    Adelman, D.D.; Stansbury, J.

    1997-12-31

    The Resource Conservation and Recovery Act (RCRA) Subtitle C, Comprehensive Environmental Response, Compensation, And Liability Act (CERCLA), and subsequent amendments have formed a comprehensive framework to deal with hazardous wastes on the national level. Key to this waste management is guidance on design (e.g., cover and bottom leachate control systems) of hazardous waste landfills. The objective of this research was to investigate the sensitivity of leachate volume at hazardous waste disposal sites to climatic, soil cover, and vegetative cover (Leaf Area Index) conditions. The computer model HELP3 which has the capability to simulate double bottom liner systems as called for in hazardous waste disposal sites was used in the analysis. HELP3 was used to model 54 combinations of climatic conditions, disposal site soil surface curve numbers, and leaf area index values to investigate how sensitive disposal site leachate volume was to these three variables. Results showed that leachate volume from the bottom double liner system was not sensitive to these parameters. However, the cover liner system leachate volume was quite sensitive to climatic conditions and less sensitive to Leaf Area Index and curve number values. Since humid locations had considerably more cover liner system leachate volume than and locations, different design standards may be appropriate for humid conditions than for and conditions.

  5. Metal-binding sites are designed to achieve optimal mechanical and signaling properties

    PubMed Central

    Dutta, Anindita; Bahar, Ivet

    2010-01-01

    Many proteins require bound metals to achieve their function. We take advantage of increasing structural data on metal-binding proteins to elucidate three properties: the involvement of metal-binding sites in the global dynamics of the protein, predicted by elastic network models, their exposure/burial to solvent, and their signal-processing properties indicated by Markovian stochastics analysis. Systematic analysis of a dataset of 145 structures reveals that the residues that coordinate metal ions enjoy remarkably efficient and precise signal transduction properties. These properties are rationalized in terms of their physical properties: participation in hinge sites that control the softest modes collectively accessible to the protein and occupancy of central positions minimally exposed to solvent. Our observations suggest that metal-binding sites may have been evolutionary selected to achieve optimum allosteric communication. They also provide insights into basic principles for designing metal-binding sites, which are verified to be met by recently designed de novo metal-binding proteins. PMID:20826340

  6. Characterization and design of the FutureGen 2.0 carbon storage site

    SciTech Connect

    Gilmore, Tyler; Bonneville, Alain; Sullivan, Charlotte; Kelley, Mark; Appriou, Delphine; Vermeul, Vince; White, Signe; Zhang, Fred; Bjornstad, Bruce; Cornet, Francois; Gerst, Jacqueline; Gupta, Neeraj; Hund, Gretchen; Horner, Jake; Last, George; Lanigan, Dave; Oostrom, Mart; McNeil, Caitlin; Moody, Mark; Rockhold, Mark; Elliott, Mike; Spane, Frank; Strickland, Chris; Swartz, Lucy; Thorne, Paul; Brown, Christopher; Hoffmann, Jeffrey; Humphreys, Kenneth

    2016-10-01

    The objective of the FutureGen 2.0 Project was to demonstrate, at the commercial scale, the technical feasibility of implementing carbon capture and storage (CCS) in a deep saline formation in Illinois, USA. Over approximately 5 years, the FutureGen Alliance conducted a detailed site-selection process and identified a site for carbon sequestration storage in Morgan County, Illinois. The storage site was fully characterized, including the collection of seismic data and the drilling and characterization of a stratigraphic borehole. The characterization data provided critical input for developing a site-specific conceptual model and subsequent numerical modeling simulations. The modeling simulations, coupled with the upstream designs of the pipeline and power plant supported the development of a detailed 90 percent design that included the injection wells and associated control and monitoring infrastructure. Collectively, all these data were used by the FutureGen Alliance to develop the required documentation to support the applications for four underground injection control (UIC) permits (one for each proposed well). In August 2014, the U.S. Environmental Protection Agency issued four, first-of-their-kind, Class VI UIC permits for carbon sequestration in the United States to the FutureGen Alliance. The information and data generated under this project have been made publically available through reports and publications, including this journal and others.

  7. Binding site elucidation and structure guided design of macrocyclic IL-17A antagonists.

    PubMed

    Liu, Shenping; Dakin, Leslie A; Xing, Li; Withka, Jane M; Sahasrabudhe, Parag V; Li, Wei; Banker, Mary Ellen; Balbo, Paul; Shanker, Suman; Chrunyk, Boris A; Guo, Zuojun; Chen, Jinshan M; Young, Jennifer A; Bai, Guoyun; Starr, Jeremy T; Wright, Stephen W; Bussenius, Joerg; Tan, Sheng; Gopalsamy, Ariamala; Lefker, Bruce A; Vincent, Fabien; Jones, Lyn H; Xu, Hua; Hoth, Lise R; Geoghegan, Kieran F; Qiu, Xiayang; Bunnage, Mark E; Thorarensen, Atli

    2016-08-16

    Interleukin-17A (IL-17A) is a principal driver of multiple inflammatory and immune disorders. Antibodies that neutralize IL-17A or its receptor (IL-17RA) deliver efficacy in autoimmune diseases, but no small-molecule IL-17A antagonists have yet progressed into clinical trials. Investigation of a series of linear peptide ligands to IL-17A and characterization of their binding site has enabled the design of novel macrocyclic ligands that are themselves potent IL-17A antagonists.

  8. Department of Energy seismic siting and design decisions: Consistent use of probabilistic seismic hazard analysis

    SciTech Connect

    Kimball, J.K.; Chander, H.

    1997-02-01

    The Department of Energy (DOE) requires that all nuclear or non-nuclear facilities shall be designed, constructed and operated so that the public, the workers, and the environment are protected from the adverse impacts of Natural Phenomena Hazards including earthquakes. The design and evaluation of DOE facilities to accommodate earthquakes shall be based on an assessment of the likelihood of future earthquakes occurrences commensurate with a graded approach which depends on the potential risk posed by the DOE facility. DOE has developed Standards for site characterization and hazards assessments to ensure that a consistent use of probabilistic seismic hazard is implemented at each DOE site. The criteria included in the DOE Standards are described, and compared to those criteria being promoted by the staff of the Nuclear Regulatory Commission (NRC) for commercial nuclear reactors. In addition to a general description of the DOE requirements and criteria, the most recent probabilistic seismic hazard results for a number of DOE sites are presented. Based on the work completed to develop the probabilistic seismic hazard results, a summary of important application issues are described with recommendations for future improvements in the development and use of probabilistic seismic hazard criteria for design of DOE facilities.

  9. On-site wastewater management system design and landslide risk assessment.

    PubMed

    Whitehead, J H; Leventhal, A

    2005-01-01

    On-site wastewater management system design and performance is significant in assessing landslide risk in areas with potential for slope instability. Much of the development in Pittwater, NSW, local government area is on steep coastal land which has a history of slope instability. Concern over cases of poorly performing or failing on-site wastewater systems and the recognition that these and newly designed systems could be contributory to slope instability has been a factor in Pittwater Council, NSW, requiring that landslide risk assessment be undertaken for new and amended on-site wastewater management systems in potentially unstable areas. This paper describes the wastewater management system design and landslide risk assessment undertaken at Pittwater Youth Hostel in accordance with the Australian Geomechanics Society's Geotechnical Risk Management procedure to comply with the Pittwater Policy. The work completed illustrates both necessary and effective interaction of the wastewater and geotechnical professions to achieve a successful outcome for the client. It is likely that this professional interaction will be increasingly common along the NSW coast and elsewhere.

  10. The Hardage Superfund Site: A remedial design/remedial action case study

    SciTech Connect

    Costello, B.; Wogsland, K.

    1997-12-31

    The Hardage Site operated as a permitted hazardous waste disposal facility from 1972 to 1980. Approximately 20 million gallons of wastes were shipped to the site. In 1984, EPA notified companies they were Potentially Responsible Parties for clean-up. NES provided project management services during RD/RA at Hardage. Remedy Design occurred between 1991 and 1993. Remedy components include 19 groundwater wells; 2,600-foot-long gravel-filled groundwater interceptor trench; 7,300-foot-deep Class-I non-hazardous injection well; removal of 3,820 drums; general site cleanup and debris removal; 80 gpm water treatment plant; and 14 acre RCRA cap over the source areas. Total construction cost was $20.7 million. The project was brought in on time and on budget with change orders less than 2% of the project total. Currently, NES is operating the remedy. The automated nature of the water treatment plant provides for 24-hour per day operation. Over 10 million gallons of water have been pumped, treated and discharged to the on-site, Class-I, non-hazardous injection well. Over 300,000 gallons of aqueous waste and NAPL have been safely removed and incinerated off-site.

  11. 77 FR 63312 - Notice of Intent: Designation of an Ocean Dredged Material Disposal Site (ODMDS) in Eastern Long...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-16

    ... AGENCY Notice of Intent: Designation of an Ocean Dredged Material Disposal Site (ODMDS) in Eastern Long... potential designation of one or more Ocean Dredged Material Disposal Sites (ODMDS) to serve the eastern Long... Environmental Management; and Rhode Island Coastal Resources Management Council. SUPPLEMENTARY INFORMATION: The...

  12. 75 FR 27783 - Decision To Evaluate a Petition To Designate a Class of Employees From the Mound Site in...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-18

    ... Doc No: 2010-11875] DEPARTMENT OF HEALTH AND HUMAN SERVICES Decision To Evaluate a Petition To Designate a Class of Employees From the Mound Site in Miamisburg, OH, To Be Included in the Special Exposure... decision to evaluate a petition to designate a class of employees from the Mound site in Miamisburg,...

  13. 77 FR 46439 - Decision To Evaluate a Petition To Designate a Class of Employees From the Baker Brothers Site in...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-03

    ... HUMAN SERVICES Decision To Evaluate a Petition To Designate a Class of Employees From the Baker Brothers Site in Toledo, Ohio, To Be Included in the Special Exposure Cohort AGENCY: National Institute for... to evaluate a petition to designate a class of employees from the Bakers Brothers site in...

  14. The Design, Synthesis, and Characterization of Open Sites on Metal Clusters

    NASA Astrophysics Data System (ADS)

    Nigra, Michael Mark

    Coordinatively unsaturated corner and edge atoms have been hypothesized to have the highest activity of sites responsible for many catalytic reactions on a metal surface. Recent studies have validated this hypothesis in varied reaction systems. However, quantification of different types of coordinatively unsaturated sites, and elucidation of their individual catalytic rates has remained a largely unresolved challenge when understanding catalysis on metal surfaces. Yet such structure-function knowledge would be invaluable to the design of more active and selective metal-surface catalysts in the future. I investigated the catalytic contributions of undercoordinated sites such as corner and edge atoms are investigated in a model reaction system using organic ligands bound to the gold nanoparticle surface. The catalyst consisted of 4 nm gold nanoparticles on a metal oxide support, using resazurin to resorufin as a model reaction system. My results demonstrate that in this system, corner atom sites are the most undercoordinated sites, and are over an order of magnitude more active when compared to undercoordinated edge atom sites, while terrace sites remain catalytically inactive for the reduction reaction of resazurin to resorufin. Catalytic activity has been also demonstrated for calixarene-bound gold nanoparticles using the reduction of 4-nitrophenol. With the 4-nitrophenol reduction reaction, a comparative study was undertaken to compare calixarene phosphine and calixarene thiol bound 4 nm gold particles. The results of the study suggested that a leached site was responsible for catalysis and not sites on the original gold nanoparticles. Future experiments with calixarene bound gold clusters could investigate ligand effects in reactions where the active site is not a leached or aggregated gold species, possibly in oxidation reactions, where electron-rich gold is hypothesized to be a good catalyst. The results that emphasize the enhanced catalytic activity of

  15. Informatics in radiology (infoRAD): HTML and Web site design for the radiologist: a primer.

    PubMed

    Ryan, Anthony G; Louis, Luck J; Yee, William C

    2005-01-01

    A Web site has enormous potential as a medium for the radiologist to store, present, and share information in the form of text, images, and video clips. With a modest amount of tutoring and effort, designing a site can be as painless as preparing a Microsoft PowerPoint presentation. The site can then be used as a hub for the development of further offshoots (eg, Web-based tutorials, storage for a teaching library, publication of information about one's practice, and information gathering from a wide variety of sources). By learning the basics of hypertext markup language (HTML), the reader will be able to produce a simple and effective Web page that permits display of text, images, and multimedia files. The process of constructing a Web page can be divided into five steps: (a) creating a basic template with formatted text, (b) adding color, (c) importing images and multimedia files, (d) creating hyperlinks, and (e) uploading one's page to the Internet. This Web page may be used as the basis for a Web-based tutorial comprising text documents and image files already in one's possession. Finally, there are many commercially available packages for Web page design that require no knowledge of HTML.

  16. 2009 Melbourne metropolitan sewerage strategy: a portfolio of decentralised and on-site concept designs.

    PubMed

    Brown, V; Jackson, D W; Khalifé, M

    2010-01-01

    The bulk and retail water companies of the greater Melbourne area are developing the 2009 Metropolitan Sewerage Strategy to provide sustainable sewerage services to 2060. The objective of the strategy is to establish long term principles and near term actions to produce a robust sewage management system for Melbourne. Melbourne's existing sewerage system is largely centralised and discharges to two major treatment plants. Several small satellite treatment plants service local urban areas generally more distant from the centralised system. Decentralised and on-site wastewater systems are options for future sewage management and could play a role in local recycling. A portfolio of 18 on-site and decentralised concept designs was developed, applicable to the full range of urban development types in Melbourne. The concepts can be used in evaluation of metropolitan system configurations as part of future integrated water cycle planning. The options included secondary and tertiary treatment systems incorporating re-use of water for non potable uses, urine separation, black and greywater separation and composting toilets. On-site and cluster treatment systems were analysed. Each option is described by its indicative capital and operating costs, energy use and water and nutrient balances. This paper summarises and compares the portfolio mix of decentralized and on-site options in Melbourne's context.

  17. Final report on the radiological surveys of designated DX firing sites at Los Alamos National Laboratory

    SciTech Connect

    1996-09-09

    CHEMRAD was contracted by Los Alamos National Laboratory to perform USRADS{reg_sign} (UltraSonic Ranging And Data System) radiation scanning surveys at designated DX Sites at the Los Alamos National Laboratory. The primary purpose of these scanning surveys was to identify the presence of Depleted Uranium (D-38) resulting from activities at the DX Firing Sites. This effort was conducted to update the most recent surveys of these areas. This current effort was initiated with site orientation on August 12, 1996. Surveys were completed in the field on September 4, 1996. This Executive Summary briefly presents the major findings of this work. The detail survey results are presented in the balance of this report and are organized by Technical Area and Site number in section 2. This organization is not in chronological order. USRADS and the related survey methods are described in section 3. Quality Control issues are addressed in section 4. Surveys were conducted with an array of radiation detectors either mounted on a backpack frame for man-carried use (Manual mode) or on a tricycle cart (RadCart mode). The array included radiation detectors for gamma and beta surface near surface contamination as well as dose rate at 1 meter above grade. The radiation detectors were interfaced directly to an USRADS 2100 Data Pack.

  18. Development of technology for the design of shallow land burial facilities at arid sites

    NASA Astrophysics Data System (ADS)

    Nyhan, J. W.; Abeele, W. V.; Drennon, B. J.; Herrera, W. J.; Lopez, E. A.; Langhorst, G. J.; Stallings, E. A.; Walker, R. D.; Martinez, J. L.

    The Los Alamos field research program involving technology development for arid shallow land burial (SLB) sites is described. Field data are presented for an integrated field experiment, which was designed to test individual SLB component experiments related to erosion control, biobarriers, and subsurface capillary and migration barriers. Field tests of biointrusion barriers at waste disposal sites and in experimental plots are reported. The results of a joint DOE/NRC experiment to evaluate leaching and transport of sorbing (Cs, Sr, Li) and nonsorbing (I, Br) solutes in sandy silt backfill are presented for steady-state and unsteady-state flow conditions. A capillary barrier experiment performed in a large caisson (3-m diameter, 6.1 m deep) is described and a year's worth of field data is presented.

  19. Autonomous Underwater Vehicle Survey Design for Monitoring Carbon Capture and Storage Sites

    NASA Astrophysics Data System (ADS)

    Bull, J. M.; Cevatoglu, M.; Connelly, D.; Wright, I. C.; McPhail, S.; Shitashima, K.

    2013-12-01

    Long-term monitoring of sub-seabed Carbon Capture and Storage (CCS) sites will require systems that are flexible, independent, and have long-endurance. In this presentation we will discuss the utility of autonomous underwater vehicles equipped with different sensor packages in monitoring storage sites. We will present data collected using Autosub AUV, as part of the ECO2 project, from the Sleipner area of the North Sea. The Autosub AUV was equipped with sidescan sonar, an EM2000 multibeam systems, a Chirp sub-bottom profiler, and a variety of chemical sensors. Our presentation will focus on survey design, and the simultaneous use of multiple sensor packages in environmental monitoring on the continental shelf.

  20. Design, Synthesis, and Biological Evaluation of Stable Colchicine Binding Site Tubulin Inhibitors as Potential Anticancer Agents

    PubMed Central

    2015-01-01

    To block the metabolically labile sites of novel tubulin inhibitors targeting the colchicine binding site based on SMART, ABI, and PAT templates, we have designed, synthesized, and biologically tested three focused sets of new derivatives with modifications at the carbonyl linker, the para-position in the C ring of SMART template, and modification of A ring of the PAT template. Structure–activity relationships of these compounds led to the identification of new benzimidazole and imidazo[4,5-c]pyridine-fused ring templates, represented by compounds 4 and 7, respectively, which showed enhanced antitumor activity and substantially improved the metabolic stability in liver microsomes compared to SMART. MOM group replaced TMP C ring and generated a potent analogue 15, which showed comparable potency to the parent SMART compound. Further modification of PAT template yielded another potent analogue 33 with 5-indolyl substituent at A ring. PMID:25122533

  1. Computational approaches for de novo design and redesign of metal-binding sites on proteins.

    PubMed

    Akcapinar, Gunseli Bayram; Sezerman, Osman Ugur

    2017-04-28

    Metal ions play pivotal roles in protein structure, function and stability. The functional and structural diversity of proteins in nature expanded with the incorporation of metal ions or clusters in proteins. Approximately one-third of these proteins in the databases contain metal ions. Many biological and chemical processes in nature involve metal ion-binding proteins, aka metalloproteins. Many cellular reactions that underpin life require metalloproteins. Most of the remarkable, complex chemical transformations are catalysed by metalloenzymes. Realization of the importance of metal-binding sites in a variety of cellular events led to the advancement of various computational methods for their prediction and characterization. Furthermore, as structural and functional knowledgebase about metalloproteins is expanding with advances in computational and experimental fields, the focus of the research is now shifting towards de novo design and redesign of metalloproteins to extend nature's own diversity beyond its limits. In this review, we will focus on the computational toolbox for prediction of metal ion-binding sites, de novo metalloprotein design and redesign. We will also give examples of tailor-made artificial metalloproteins designed with the computational toolbox.

  2. Early Site Permit Demonstration Program: Guidelines for determining design basis ground motions. Volume 2, Appendices

    SciTech Connect

    Not Available

    1993-03-18

    This report develops and applies a methodology for estimating strong earthquake ground motion. The motivation was to develop a much needed tool for use in developing the seismic requirements for structural designs. An earthquake`s ground motion is a function of the earthquake`s magnitude, and the physical properties of the earth through which the seismic waves travel from the earthquake fault to the site of interest. The emphasis of this study is on ground motion estimation in Eastern North America (east of the Rocky Mountains), with particular emphasis on the Eastern United States and southeastern Canada. Eastern North America is a stable continental region, having sparse earthquake activity with rare occurrences of large earthquakes. While large earthquakes are of interest for assessing seismic hazard, little data exists from the region to empirically quantify their effects. The focus of the report is on the attributes of ground motion in Eastern North America that are of interest for the design of facilities such as nuclear power plants. This document, Volume II, contains Appendices 2, 3, 5, 6, and 7 covering the following topics: Eastern North American Empirical Ground Motion Data; Examination of Variance of Seismographic Network Data; Soil Amplification and Vertical-to-Horizontal Ratios from Analysis of Strong Motion Data From Active Tectonic Regions; Revision and Calibration of Ou and Herrmann Method; Generalized Ray Procedure for Modeling Ground Motion Attenuation; Crustal Models for Velocity Regionalization; Depth Distribution Models; Development of Generic Site Effects Model; Validation and Comparison of One-Dimensional Site Response Methodologies; Plots of Amplification Factors; Assessment of Coupling Between Vertical & Horizontal Motions in Nonlinear Site Response Analysis; and Modeling of Dynamic Soil Properties.

  3. Remedial action plan and site design for stabilization of the inactive uranium mill tailings sites at Rifle, Colorado

    SciTech Connect

    Not Available

    1990-02-01

    This volume contains appendices D6 through D8 containing laboratory test data: from MK-F investigation, 1987, Old Rifle and New Rifle sites; on bentonite amended radon barrier material; and from MK-F investigation, 1987, riprap tests.

  4. Modifications to the remedial action plan and site design for stabilization of the inactive Uranium Mill Tailings Site at Green River, Utah

    SciTech Connect

    Not Available

    1994-09-01

    Modifications to the water resources protection strategy detailed in the remedial action plan for the Green River, Utah, disposal site are presented. The modifications are based on new information, including ground water quality data collected after remedial action was completed and on a revised assessment of disposal cell design features, surface conditions, and site hydrogeology. The modifications will result in compliance with the U.S. EPA proposed ground water standards (52 FR 36000 (1987)).

  5. Postclosure performance assessment of the SCP (Site Characterization Plan) conceptual design for horizontal emplacement: Revision 1

    SciTech Connect

    Not Available

    1987-08-01

    This report is a preliminary postclosure performance assessment of the repository design specified in the Site Characterization Plan Conceptual Design Report (SCP-CDR) for horizontal emplacement of high-level nuclear waste. At the time that these analyses were done, horizontal emplacement was the preferred orientation for the waste packages but vertical emplacement is now the reference design. This assessment consists of (1) a review of the regulatory requirements and strategy to demonstrate compliance with these requirements, (2) an analysis of the performance of the total repository system, (3) an analysis of the thermomechanical behavior of the repository, (4) an analysis of brine mobility in the repository, (5) an analysis of the waste package performance, (6) an analysis of the performance of seals, and (7) comments on the sensitivity of the various performance measures to uncertainties in the data and models. These are preliminary analyses and, in most cases, involve bounding calculations of the repository behavior. They have several purposes including (1) assessing how well this conceptual design ''measures up'' against requirements, (2) gaining experience in implementing the performance assessment strategy and tools and thereby learning where improvements are needed, (3) helping to identify needed data, and (4) helping to indicate required design modifications. 26 refs., 40 figs., 20 tabs.

  6. Investigation into the optimal hydrologic design of porous concrete sites using mathematical modeling

    NASA Astrophysics Data System (ADS)

    Syrrakou, C.; Fitch, J.; Eliassen, T.; Ahearn, W.; Pinder, G. F.

    2011-12-01

    Increase in the amount of paved areas as a result of urbanization in modern societies has lead to the need of stormwater best management practices (BMPs). In that direction, porous pavement has been used successfully in regions of warm climate and application in regions of colder climate is an object of ongoing research with encouraging results to date. The significant cost and effort that accompanies the maintenance of porous pavement facilities calls for a design tool that can be used prior construction to facilitate the design process and also post production to evaluate the site's overall performance. Such a tool is a mathematical model which takes into account the different physical processes that can occur in a porous concrete system including recharge from rainfall, runoff from surrounding conventionally paved areas, vertical flow, storage and finally infiltration into the subsurface. In this research a three-dimensional saturated-unsaturated flow and transport model is modified to account for flow through the porous concrete slab and also evaporation. Runoff is accounted by means of a two-dimensional surface flow model which calculates the infiltration into the perimeter porous concrete area. The mathematical model is used to simulate a porous concrete site which operates as a public parking lot facility in Randolph, Vermont. The subgrade soil in the area of interest consists mainly of dense till deposits typically found in New England. Such deposits can result in small infiltration rates. The specific site is unique not only in terms of the underlying geology but also the heavy instrumentation not usually observed in similar sites. The instrumentation includes a number of groundwater wells which are being monitored continuously through a pressure transducer system, temperature probes installed inside the porous concrete and a detailed underdrain system located in the porous concrete's sub-base accumulating infiltrated water. Laboratory research is also

  7. Advanced Imaging for Glaucoma Study: Design, Baseline Characteristics, and Inter-Site Comparison

    PubMed Central

    Le, Phuc V.; Zhang, Xinbo; Francis, Brian A.; Varma, Rohit; Greenfield, David S.; Schuman, Joel S.; Loewen, Nils; Huang, David

    2014-01-01

    Purpose To report the baseline characteristics of the participants in the Advanced Imaging for Glaucoma Study. To compare the participating sites for variations among subjects and the performance of imaging instruments. Design Multi-center longitudinal observational cohort study Methods A total of 788 participants (1,329 eyes) were enrolled from three academic referral centers. There were 145 participants (289 eyes) in the normal group, 394 participants (663 eyes) in the glaucoma suspect/preperimetric glaucoma group, and 249 participants (377 eyes) in the perimetric glaucoma group. Participants underwent a full clinical exam, standard automated perimetry, and imaging with time-domain and Fourier-domain optical coherence tomography (OCT), scanning laser polarimetry, and confocal scanning laser ophthalmoscopy. Main Outcome Measures The baseline average, population standard deviation, and repeatability of imaging-derived anatomic variables were reported for each technology and center. Results Compared to the normal participants, glaucoma suspect/preperimetric glaucoma and perimetric glaucoma groups had significantly reduced anatomic measurements. Repeatability of nerve fiber layer thickness was best for Fourier-domain OCT (overall coefficient of variation < 2%), followed by time-domain OCT (coefficient of variation 2-2.9%), scanning laser polarimetry (coefficient of variation 2.6-4.5%), and confocal scanning laser ophthalmoscopy rim area (coefficient of variation 4.2-7.6%). A mixed-effects model showed that the differences between sites was less than 25 percent of the variation within groups and less than the differences between the normal and glaucoma suspect/preperimetric glaucoma group. Conclusions Site-to-site variation was smaller than both the variation within groups and the changes due to glaucoma. Therefore pooling of participants between sites is appropriate. PMID:25447111

  8. Uncertainty based optimal monitoring network design for a chlorinated hydrocarbon contaminated site.

    PubMed

    Chadalavada, Sreenivasulu; Datta, Bithin; Naidu, Ravi

    2011-02-01

    An application of a newly developed optimal monitoring network for the delineation of contaminants in groundwater is demonstrated in this study. Designing a monitoring network in an optimal manner helps to delineate the contaminant plume with a minimum number of monitoring wells at optimal locations at a contaminated site. The basic principle used in this study is that the wells are installed where the measurement uncertainties are minimum at the potential monitoring locations. The development of the optimal monitoring network is based on the utilization of contaminant concentration data from an existing initial arbitrary monitoring network. The concentrations at the locations that were not sampled in the study area are estimated using geostatistical tools. The uncertainty in estimating the contaminant concentrations at such locations is used as design criteria for the optimal monitoring network. The uncertainty in the study area was quantified by using the concentration estimation variances at all the potential monitoring locations. The objective function for the monitoring network design minimizes the spatial concentration estimation variances at all potential monitoring well locations where a monitoring well is not to be installed as per the design criteria. In the proposed methodology, the optimal monitoring network is designed for the current management period and the contaminant concentration data estimated at the potential observation locations are then used as the input to the network design model. The optimal monitoring network is designed for the consideration of two different cases by assuming different initial arbitrary existing data. Three different scenarios depending on the limit of the maximum number of monitoring wells that can be allowed at any period are considered for each case. In order to estimate the efficiency of the developed optimal monitoring networks, mass estimation errors are compared for all the three different scenarios of the two

  9. Binding site elucidation and structure guided design of macrocyclic IL-17A antagonists

    PubMed Central

    Liu, Shenping; Dakin, Leslie A.; Xing, Li; Withka, Jane M.; Sahasrabudhe, Parag V.; Li, Wei; Banker, Mary Ellen; Balbo, Paul; Shanker, Suman; Chrunyk, Boris A.; Guo, Zuojun; Chen, Jinshan M.; Young, Jennifer A.; Bai, Guoyun; Starr, Jeremy T.; Wright, Stephen W.; Bussenius, Joerg; Tan, Sheng; Gopalsamy, Ariamala; Lefker, Bruce A.; Vincent, Fabien; Jones, Lyn H.; Xu, Hua; Hoth, Lise R.; Geoghegan, Kieran F.; Qiu, Xiayang; Bunnage, Mark E.; Thorarensen, Atli

    2016-01-01

    Interleukin-17A (IL-17A) is a principal driver of multiple inflammatory and immune disorders. Antibodies that neutralize IL-17A or its receptor (IL-17RA) deliver efficacy in autoimmune diseases, but no small-molecule IL-17A antagonists have yet progressed into clinical trials. Investigation of a series of linear peptide ligands to IL-17A and characterization of their binding site has enabled the design of novel macrocyclic ligands that are themselves potent IL-17A antagonists. PMID:27527709

  10. Design and Construction of Deinococcus Radiodurans for Biodegradation of Organic Toxins at Radioactive DOE Waste Sites

    SciTech Connect

    Michael J. Daly; Lawrence P. Wackett; James K. Fredrickson

    2001-04-22

    Seventy million cubic meters of ground and three trillion liters of groundwater have been contaminated by leaking radioactive waste generated in the United States during the Cold War. A cleanup technology is being developed based on the extremely radiation resistant bacterium Deinococcus radiodurans that is being engineered to express bioremediating functions. Research aimed at developing D. radiodurans for organic toxin degradation in highly radioactive waste sites containing radionuclides, heavy metals, and toxic organic compounds was started by this group.Work funded by the existing grant has already contributed to eleven papers on the fundamental biology of D. radiodurans and its design for bioremediation of highly radioactive waste environments

  11. Remedial Action Plan and site design for stabilization of the inactive uranium mill tailings site at Gunnison, Colorado. Attachment 2, Geology report: Final report

    SciTech Connect

    Not Available

    1992-10-01

    Detailed investigations of geologic, geomorphic, and seismic conditions at the Landfill disposal site were conducted. The purpose of these studies was basic site characterization and identification of potential geologic hazards that could affect long-term site stability. Subsequent engineering studies, such as analyses of hydrologic and liquefaction hazards, used the data developed in these studies. The geomorphic analysis was employed in the design of effective erosion protection. Studies of the regional and local seismotectonic setting, which included a detailed search for possible capable faults within a 65-kilometer (km) (40-mile) radius of the site, provided the basis for seismic design parameters. The scope of work performed included the following: Compilation and analysis of previous published and unpublished geologic literature and maps. Review of historical and instrumental earthquake data. Review of site-specific subsurface geologic data, including lithologic and geophysical logs of exploratory boreholes advanced in the site area. Photogeologic interpretations of existing conventional aerial photographs. Ground reconnaissance and mapping of the site region.

  12. Remedial Action Plan and site design for stabilization of the inactive uranium mill tailings site at Durango, Colorado: Attachment 2, Geology report. Revised final report

    SciTech Connect

    Not Available

    1991-12-01

    Detailed investigations of geologic, geomorphic, and seismic conditions at the Bodo Canyon disposal site were conducted. The purpose of these investigations was basic site characterization and identification of potential geologic hazards that could affect long-term site stability. Subsequent engineering studies, such as analyses of hydrologic and liquefaction hazards, used the data developed in these studies. The geomorphic analysis was employed in the design of effective erosion protection. Studies of the regional and local seismotectonic setting, which included a detailed search for possible capable faults within a 65 kilometer radius of the site, provided the basis for seismic design parameters. The scope of work performed included the following: Compilation and analysis of previous published and unpublished geologic literature and maps; Review of historical and instrumental earthquake data; Review of site-specific subsurface geologic data, including lithologic and geophysical logs of exploratory boreholes advanced in the site area; Photogeologic interpretations of existing conventional aerial photographs; and, Ground reconnaissance and mapping of the site region.

  13. Computational design of a red fluorophore ligase for site-specific protein labeling in living cells

    SciTech Connect

    Liu, Daniel S.; Nivon, Lucas G.; Richter, Florian; Goldman, Peter J.; Deerinck, Thomas J.; Yao, Jennifer Z.; Richardson, Douglas; Phipps, William S.; Ye, Anne Z.; Ellisman, Mark H.; Drennan, Catherine L.; Baker, David; Ting, Alice Y.

    2014-10-13

    In this study, chemical fluorophores offer tremendous size and photophysical advantages over fluorescent proteins but are much more challenging to target to specific cellular proteins. Here, we used Rosetta-based computation to design a fluorophore ligase that accepts the red dye resorufin, starting from Escherichia coli lipoic acid ligase. X-ray crystallography showed that the design closely matched the experimental structure. Resorufin ligase catalyzed the site-specific and covalent attachment of resorufin to various cellular proteins genetically fused to a 13-aa recognition peptide in multiple mammalian cell lines and in primary cultured neurons. We used resorufin ligase to perform superresolution imaging of the intermediate filament protein vimentin by stimulated emission depletion and electron microscopies. This work illustrates the power of Rosetta for major redesign of enzyme specificity and introduces a tool for minimally invasive, highly specific imaging of cellular proteins by both conventional and superresolution microscopies.

  14. Computational design of a red fluorophore ligase for site-specific protein labeling in living cells

    PubMed Central

    Liu, Daniel S.; Nivón, Lucas G.; Richter, Florian; Goldman, Peter J.; Deerinck, Thomas J.; Yao, Jennifer Z.; Richardson, Douglas; Phipps, William S.; Ye, Anne Z.; Ellisman, Mark H.; Drennan, Catherine L.; Baker, David; Ting, Alice Y.

    2014-01-01

    Chemical fluorophores offer tremendous size and photophysical advantages over fluorescent proteins but are much more challenging to target to specific cellular proteins. Here, we used Rosetta-based computation to design a fluorophore ligase that accepts the red dye resorufin, starting from Escherichia coli lipoic acid ligase. X-ray crystallography showed that the design closely matched the experimental structure. Resorufin ligase catalyzed the site-specific and covalent attachment of resorufin to various cellular proteins genetically fused to a 13-aa recognition peptide in multiple mammalian cell lines and in primary cultured neurons. We used resorufin ligase to perform superresolution imaging of the intermediate filament protein vimentin by stimulated emission depletion and electron microscopies. This work illustrates the power of Rosetta for major redesign of enzyme specificity and introduces a tool for minimally invasive, highly specific imaging of cellular proteins by both conventional and superresolution microscopies. PMID:25313043

  15. Computational design of a red fluorophore ligase for site-specific protein labeling in living cells

    DOE PAGES

    Liu, Daniel S.; Nivon, Lucas G.; Richter, Florian; ...

    2014-10-13

    In this study, chemical fluorophores offer tremendous size and photophysical advantages over fluorescent proteins but are much more challenging to target to specific cellular proteins. Here, we used Rosetta-based computation to design a fluorophore ligase that accepts the red dye resorufin, starting from Escherichia coli lipoic acid ligase. X-ray crystallography showed that the design closely matched the experimental structure. Resorufin ligase catalyzed the site-specific and covalent attachment of resorufin to various cellular proteins genetically fused to a 13-aa recognition peptide in multiple mammalian cell lines and in primary cultured neurons. We used resorufin ligase to perform superresolution imaging of themore » intermediate filament protein vimentin by stimulated emission depletion and electron microscopies. This work illustrates the power of Rosetta for major redesign of enzyme specificity and introduces a tool for minimally invasive, highly specific imaging of cellular proteins by both conventional and superresolution microscopies.« less

  16. Oxidative stress alters base excision repair pathway and increases apoptotic response in Apurinic/apyrimidinic endonuclease 1/Redox factor-1 haploinsufficient mice

    PubMed Central

    Unnikrishnan, Archana; Raffoul, Julian J.; Patel, Hiral V.; Prychitko, Thomas M.; Anyangwe, Njwen; Meira, Lisiane B.; Friedberg, Errol C.; Cabelof, Diane C.; Heydari, Ahmad R.

    2009-01-01

    Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) is the redox regulator of multiple stress-inducible transcription factors, such as NF-κB, and the major 5’-endonuclease in base excision repair (BER). We utilized mice containing heterozygous gene-targeted deletion of APE1/Ref-1 (Apex+/-) to determine the impact of APE1/Ref-1 haploinsufficiency on the processing of oxidative DNA damage induced by 2-nitropropane (2-NP) in the liver tissue of mice. APE1/Ref-1 haploinsufficiency results in a significant decline in NF-κB DNA binding activity in response to oxidative stress in liver. In addition, loss of APE1/Ref-1 increases the apoptotic response to oxidative stress where a significant increase in GADD45g expression, p53 protein stability and caspase activity are observed. Oxidative stress displays a differential impact on monofunctional (UDG) and bifunctional (OGG1) DNA glycosylase initiated BER in liver of Apex+/- mice. APE1/Ref-1 haploinsufficiency results in a significant decline in the repair of oxidized bases (e.g., 8-OHdG), while removal of uracil is increased in liver nuclear extracts of mice using an in vitro BER assay. Apex+/- mice exposed to 2-NP displayed a significant decline in 3’-OH-containing single-strand breaks and an increase in aldehydic lesions in their liver DNA suggesting an accumulation of repair intermediates of failed bifunctional DNA glycosylase initiated BER. PMID:19268524

  17. 41 CFR 102-76.20 - What issues must Federal agencies consider in providing site planning and landscape design services?

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ...) FEDERAL MANAGEMENT REGULATION REAL PROPERTY 76-DESIGN AND CONSTRUCTION Design and Construction § 102-76.20... 41 Public Contracts and Property Management 3 2014-01-01 2014-01-01 false What issues must Federal agencies consider in providing site planning and landscape design services? 102-76.20 Section...

  18. 75 FR 39523 - Notice of Intent: Designation of an Ocean Dredged Material Disposal Site (ODMDS) Off the Mouth of...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-09

    ... AGENCY Notice of Intent: Designation of an Ocean Dredged Material Disposal Site (ODMDS) Off the Mouth of... Intent to prepare an Environmental Impact Statement (EIS) for the designation of an ODMDS off the mouth... an EIS to designate a new ODMDS offshore the mouth of the St. Johns River. The EIS will provide the...

  19. Remedial Action Plan and site conceptual design for stabilization of the inactive uranium mill tailings site at Spook, Wyoming

    SciTech Connect

    Matthews, M L; Sullivan, M

    1990-04-01

    This Remedial Action Plan (RAP) has been developed to serve a threefold purpose. It presents the series of activities which are proposed by the US Department of Energy (DOE) to accomplish long-term stabilization and control of radioactive materials at an inactive uranium processing site northeast of Casper, Wyoming, and referred to as the Spook site. It provides a characterization of the present conditions at the site and also serves to document the concurrence of the State of Wyoming and the US Nuclear Regulatory Commission (NRC) in the remedial action. This agreement, upon execution by the DOE and the State of Wyoming, and concurrence by the NRC, becomes Appendix B of the Cooperative Agreement.

  20. Design and identification of a high efficient formic acid cleavage site for separation of fusion protein.

    PubMed

    Zhang, Huaguang; Li, Mei; Shi, Shuangfeng; Yin, Chao; Jia, Shirong; Wang, Zhixing; Liu, Yuhui

    2015-02-01

    The release of target protein with high efficiency and low cost from expressed fusion protein is a key requirement for commercial production of target proteins. To establish such a cleavage system, we have designed four formic acid (FA) cleavage sites C1 (DPDPDP), C2 (DPPDPP), C3 (DDDDPI) and C4 (IVDPNP), which was placed in between the E and G fusion protein. Four expression vectors were individually constructed and expressed in Escherichia coli. Purified proteins were reacted with a series of FA concentrations or under different temperatures followed by SDS-PAGE gel electrophoresis to verify the degree of cleavage efficiency. Results showed that the C2 was the most efficient site compared with the other three. After optimization of cleavage conditions for E-C2-G, the cleavage efficiently could reach as high as 87.3% within 2.5 h in 37% FA at 45 °C. Comparing with previous reports, a significant reduction (26%) of FA concentration at a lower temperature in a short duration of reaction (18 times less) was achieved. We believe the cleavage site of DPPDPP identified in this study can be used in the large-scale production of valuable fusion proteins to save the cost, time and energy.

  1. Structure-based drug design: exploring the proper filling of apolar pockets at enzyme active sites.

    PubMed

    Zürcher, Martina; Diederich, François

    2008-06-20

    The proper filling of apolar pockets at enzyme active sites is central for increasing binding activity and selectivity of hits and leads in medicinal chemistry. In our structure-based design approach toward the generation of potent enzyme inhibitors, we encountered a variety of challenges in gaining suitable binding affinity from the occupation of such pockets. We summarize them here for the first time. A fluorine scan of tricyclic thrombin inhibitors led to the discovery of favorable orthogonal dipolar C-F...CO interactions. Efficient cation-pi interactions were established in the S4 pocket of factor Xa, another serine protease from the blood coagulation cascade. Changing from mono- to bisubstrate inhibitors of catechol O-methyltransferase, a target in the L-Dopa-based treatment of Parkinson's disease, enabled the full exploitation of a previously unexplored hydrophobic pocket. Conformational preorganization of a pocket at an enzyme active site is crucial for harvesting binding affinity. This is demonstrated for two enzymes from the nonmevalonate pathway of isoprenoid biosynthesis, IspE and IspF, which are pursued as antimalarial targets. Disrupting crystallographically defined water networks on the way into a pocket might cost all of the binding free enthalpy gained from its occupation, as revealed in studies with tRNA-guanine transglycosylase, a target against shigellosis. Investigations of the active site of plasmepsin II, another antimalarial target, showed that principles for proper apolar cavity filling, originally developed for synthetic host-guest systems, are also applicable to enzyme environments.

  2. The water network in galectin-3 ligand binding site guides inhibitor design.

    PubMed

    Su, Jiyong; Zhang, Tao; Wang, Peiqi; Liu, Fengjian; Tai, Guihua; Zhou, Yifa

    2015-03-01

    Galectin-3 (Gal-3) which shows affinity of β-galactosides is a cancer-related protein. Thus, it is important to understand its ligand binding mechanism and then design its specific inhibitor. It was suggested that the positions of water molecules in Gal-3 ligand-binding site could be replaced by appropriate chemical groups of ideal inhibitors. However, the reported structures of Gal-3 carbohydrate recognition domain (CRD) complexed with lactose showed that the number of water molecules are different and the water positions are inconsistent in the ligand-binding site. This study reported four high-resolution (1.24-1.19 Å) structures of Gal-3 CRD complexed with lactose, and accurately located 12 conserved water molecules in the water network of Gal-3 CRD ligand-binding site by merging these structures. These water molecules either directly stabilize the binding of Gal-3 CRD and lactose, or hold the former water molecules at the right place. In particular, water molecule 4 (W4) which only coordinates with water molecule 5 (W5) and water molecule 6 (W6) plays a key role in stabilizing galactose residue. In addition, by three-dimensional alignment of the positions of all residues, 14 flexible parts of Gal-3 CRD were found to dynamically fluctuate in the crystalline environment.

  3. Health Literacy Online: A Guide to Writing and Designing Easy-to-Use Health Web Sites.

    PubMed

    Hou, Su-I

    2012-09-01

    Health Literacy Online: A Guide to Writing and Designing Easy-to-Use Health Web Sites is a practical and well-written resource for public health and health communication professionals and web designers. This guide builds on the principles of web usability and adds to existing best practices by providing research-based strategies for writing and designing health websites especially for users with limited literacy and health literacy skills. This guide synthesizes years of lessons learned from Office of Disease Prevention and Health Promotion's original research with hundreds of web users, experience with revising the healthfinder.gov, as well as strategies supported by the Research-Based Web Design and Usability Guidelines (Usability.gov). In the United States, roughly one third of adults have limited literacy skills, yet far more (as many as 90%) have limited health literacy skills, meaning they have trouble understanding complex health information. This how-to guide is timely and developed with the aim of creating easy-to-use health websites to reach as many web users as possible, especially those with limited literacy and health literacy skills.

  4. Rational Design of Dual Active Sites in a Single Protein Scaffold: A Case Study of Heme Protein in Myoglobin.

    PubMed

    Shu, Xiao-Gang; Su, Ji-Hu; Du, Ke-Jie; You, Yong; Gao, Shu-Qin; Wen, Ge-Bo; Tan, Xiangshi; Lin, Ying-Wu

    2016-06-01

    Rational protein design has been proven to be a powerful tool for creating functional artificial proteins. Although many artificial metalloproteins with a single active site have been successfully created, those with dual active sites in a single protein scaffold are still relatively rare. In this study, we rationally designed dual active sites in a single heme protein scaffold, myoglobin (Mb), by retaining the native heme site and creating a copper-binding site remotely through a single mutation of Arg118 to His or Met. Isothermal titration calorimetry (ITC) and electron paramagnetic resonance (EPR) studies confirmed that a copper-binding site of [3-His] or [2-His-1-Met] motif was successfully created in the single mutant of R118H Mb and R118M Mb, respectively. UV/Vis kinetic spectroscopy and EPR studies further revealed that both the heme site and the designed copper site exhibited nitrite reductase activity. This study presents a new example for rational protein design with multiple active sites in a single protein scaffold, which also lays the groundwork for further investigation of the structure and function relationship of heme/non-heme proteins.

  5. Structure-guided inhibitor design for human FAAH by interspecies active site conversion.

    PubMed

    Mileni, Mauro; Johnson, Douglas S; Wang, Zhigang; Everdeen, Daniel S; Liimatta, Marya; Pabst, Brandon; Bhattacharya, Keshab; Nugent, Richard A; Kamtekar, Satwik; Cravatt, Benjamin F; Ahn, Kay; Stevens, Raymond C

    2008-09-02

    The integral membrane enzyme fatty acid amide hydrolase (FAAH) hydrolyzes the endocannabinoid anandamide and related amidated signaling lipids. Genetic or pharmacological inactivation of FAAH produces analgesic, anxiolytic, and antiinflammatory phenotypes but not the undesirable side effects of direct cannabinoid receptor agonists, indicating that FAAH may be a promising therapeutic target. Structure-based inhibitor design has, however, been hampered by difficulties in expressing the human FAAH enzyme. Here, we address this problem by interconverting the active sites of rat and human FAAH using site-directed mutagenesis. The resulting humanized rat (h/r) FAAH protein exhibits the inhibitor sensitivity profiles of human FAAH but maintains the high-expression yield of the rat enzyme. We report a 2.75-A crystal structure of h/rFAAH complexed with an inhibitor, N-phenyl-4-(quinolin-3-ylmethyl)piperidine-1-carboxamide (PF-750), that shows strong preference for human FAAH. This structure offers compelling insights to explain the species selectivity of FAAH inhibitors, which should guide future drug design programs.

  6. A reliable facility location design model with site-dependent disruption in the imperfect information context.

    PubMed

    Yun, Lifen; Wang, Xifu; Fan, Hongqiang; Li, Xiaopeng

    2017-01-01

    This paper proposes a reliable facility location design model under imperfect information with site-dependent disruptions; i.e., each facility is subject to a unique disruption probability that varies across the space. In the imperfect information contexts, customers adopt a realistic "trial-and-error" strategy to visit facilities; i.e., they visit a number of pre-assigned facilities sequentially until they arrive at the first operational facility or give up looking for the service. This proposed model aims to balance initial facility investment and expected long-term operational cost by finding the optimal facility locations. A nonlinear integer programming model is proposed to describe this problem. We apply a linearization technique to reduce the difficulty of solving the proposed model. A number of problem instances are studied to illustrate the performance of the proposed model. The results indicate that our proposed model can reveal a number of interesting insights into the facility location design with site-dependent disruptions, including the benefit of backup facilities and system robustness against variation of the loss-of-service penalty.

  7. Development and application of site mapping methods for the design of glycosaminoglycans.

    PubMed

    Agostino, Mark; Gandhi, Neha S; Mancera, Ricardo L

    2014-09-01

    Glycosaminoglycans (GAGs) are complex polysaccharides involved in a wide range of biological signaling events, as well as being important as biological structural materials. Despite the ubiquity and importance of GAG-protein interactions in biological systems and potentially as therapeutic targets, detailed structures of such interactions are sparse in availability. Computational methods can provide detailed structural knowledge of these interactions; however, they should be evaluated against suitable test systems prior to their widespread use. In this study, we have investigated the application of automated molecular docking and interaction mapping techniques to characterizing GAG-protein interactions. A series of high-resolution X-ray crystal structures of GAGs in complex with proteins was used to evaluate the approaches. Accurately scoring the pose fitting best with the crystal structure was a challenge for all docking programs evaluated. The site mapping technique offered excellent prediction of the key residues involved in ligand recognition, comparable to the best pose and improved over the top-ranked pose. A design protocol incorporating site- and ligand-based mapping techniques was developed and applied to identify GAGs capable of binding to acidic fibroblast growth factor (aFGF). The protocol was able to identify ligands known to bind to aFGF and accurately able to predict the binding modes of those ligands when using a known ligand-binding conformation of the protein. This study demonstrates the value of mapping-based techniques in identifying specific GAG epitopes recognized by proteins and for GAG-based drug design.

  8. A reliable facility location design model with site-dependent disruption in the imperfect information context

    PubMed Central

    Yun, Lifen; Wang, Xifu; Fan, Hongqiang; Li, Xiaopeng

    2017-01-01

    This paper proposes a reliable facility location design model under imperfect information with site-dependent disruptions; i.e., each facility is subject to a unique disruption probability that varies across the space. In the imperfect information contexts, customers adopt a realistic “trial-and-error” strategy to visit facilities; i.e., they visit a number of pre-assigned facilities sequentially until they arrive at the first operational facility or give up looking for the service. This proposed model aims to balance initial facility investment and expected long-term operational cost by finding the optimal facility locations. A nonlinear integer programming model is proposed to describe this problem. We apply a linearization technique to reduce the difficulty of solving the proposed model. A number of problem instances are studied to illustrate the performance of the proposed model. The results indicate that our proposed model can reveal a number of interesting insights into the facility location design with site-dependent disruptions, including the benefit of backup facilities and system robustness against variation of the loss-of-service penalty. PMID:28486564

  9. Remedial Action Plan and Site Design for Stabilization of the Inactive Uranium Mill Tailings Site, Maybell, Colorado. Remedial action selection report: Attachment 2, Geology report, Final

    SciTech Connect

    Not Available

    1994-06-01

    The Maybell uranium mill tailings site is 25 miles (mi) (40 kilometers [km]) west of the town of Craig, Colorado, in Moffat County, in the northwestern part of the state. The unincorporated town of Maybell is 5 road mi (8 km) southwest of the site. The designated site covers approximately 110 acres (ac) (45 hectares [ha]) and consists of a concave-shaped tailings pile and rubble from the demolition of the mill buildings buried in the former mill area. Contaminated materials at the Maybell processing site include the tailings pile, which has an average depth of 20 feet (ft) (6 meters [m]) and contains 2.8 million cubic yards (yd{sup 3}) (2.1 million cubic meters [m{sup 3}]) of tailings. The former mill processing area is on the north side of the site and contains 20,000 yd{sup 3} (15,000 m{sup 3}) of contaminated demolition debris. Off-pile contamination is present and includes areas adjacent to the tailings pile, as well as contamination dispersed by wind and surface water flow. The volume of off-pile contamination to be placed in the disposal cell is 550,000 yd{sup 3} (420,000 m{sup 3}). The total volume of contaminated materials to be disposed of as part of the remedial action is estimated to be 3.37 million yd{sup 3} (2.58 million m{sup 3}). Information presented in this Final Remedial Action Plan (RAP) and referenced in supporting documents represents the current disposal cell design features and ground water compliance strategy proposed by the US Department of Energy (DOE) for the Maybell, Colorado, tailings site. Both the disposal cell design and the ground water compliance strategy have changed from those proposed prior to the preliminary final RAP document as a result of prudent site-specific technical evaluations.

  10. The Importance of Heuristic Evaluation and Usability Testing in the User Interface Design for a Family Health History Web Site

    PubMed Central

    Kinzie, Mable; Cohn, Wendy; Knaus, William

    2001-01-01

    In order to develop the most effective health information web sites or instructional materials for physicians and consumers, designers frequently use a suite of best design practices, including Needs Assessment, Goal Analysis, User Interface Principles, and Rapid Prototyping. These best practices are not enough, however. If designers want to ensure the best possible product, they must also subject their products to Heuristic Evaluation and Usability Testing. A Web site designed to collect and evaluate family health history will be used to illustrate these principles.

  11. Creating patient and family education web sites: design and content of the home parenteral nutrition family caregivers web site.

    PubMed

    Fitzgerald, Sharon A; Macan Yadrich, Donna; Werkowitch, Marilyn; Piamjariyakul, Ubolrat; Smith, Carol E

    2011-11-01

    When managing chronic illnesses, caregivers repeatedly seek online information about providing complex, long-term care but often neglect to find information about how to care for themselves. Poor health among caregivers is not only detrimental to their own well-being but may also result in harm to those for whom they care. For this reason, caregivers need access to information and activities about caring for themselves in addition to the information about managing home care they are already likely to seek. The HPN Family Caregivers Web site was developed to guide caregivers through the process of caring for themselves by establishing a caregiving routine, self-monitoring their mental and physical health, and practicing good sleep hygiene, while also managing the complexities of home care. While Web site information, activities, and algorithms for managing chronic illnesses need to be specific to each population, the content guiding caregivers to care for their own health is universal.

  12. Houdini: Site and locomotion analysis-driven design of an in-tank mobile cleanup robot

    SciTech Connect

    Schempf, H.

    1995-10-01

    This paper describes design and locomotion analysis efforts to develop a new reconfigurable and collapsible working machine, dubbed Houdini, to remotely clean up hazardous-waste and petroleum storage tanks. The tethered robot system is designed to allow remote entry through man-way openings as small as 0.61 m in diameter, after which it expands its locomotors and opens up its collapsible backhoe/manipulator and plow to subsequently perform waste or material handling operations. The design is optimized to meet stringent site and safety requirements, and represents a viable alternative to (1) the long-reach manipulation systems proposed for hazardous storage tank cleanup, and (2) confined-entry manual cleanup approaches. The system development has been funded to provide waste mobilization and removal solutions for the hazardous waste storage tanks in the Department of Energy (DoE) Fernald and Oak Ridge complexes. Other potential applications areas are the cleanup of heavy-crude petroleum storage tanks. The author has developed a fully operational prototype which is currently undergoing testing.

  13. Fly ash design manual for road and site applications. Volume 2: Slurried placement: Final report

    SciTech Connect

    DiGioia, A.M. Jr.; Brendel, G.F.; McLaren, R.J.; Balsamo, N.J.; Glogowski, P.E.; Kelley, J.M.

    1992-04-01

    This design manual is the second of two volumes pertaining to the use of fly ash as a construction material in road and site development applications. Volume 1 describes the use of fly ash conditioned with small amounts of water to produce a solid, soil-like material. Volume 2 describes the use of fly ash mixed with sufficient water to create a fluid or semi-fluid material. In most applications, Portland cement is added to the mixture to provide the desired level of strength. In some applications, aggregate filler is also added to the mixture to improve the durability of the mix. The flowable nature of these materials determines the manner in which they are handled, placed, and used in construction. The following uses are discussed: Slurry Backfills; Grouting; and Hydraulic Fills. Miscellaneous uses are also discussed, including the use of fly ash in high fly ash content concrete, unsintered synthetic aggregate, and slurry walls. Also included are three appendices containing additional information of interest to the designer: Mix Design; Laboratory Test Procedures; and a Bibliography.

  14. Designing artificial 2D crystals with site and size controlled quantum dots.

    PubMed

    Xie, Xuejun; Kang, Jiahao; Cao, Wei; Chu, Jae Hwan; Gong, Yongji; Ajayan, Pulickel M; Banerjee, Kaustav

    2017-08-30

    Ordered arrays of quantum dots in two-dimensional (2D) materials would make promising optical materials, but their assembly could prove challenging. Here we demonstrate a scalable, site and size controlled fabrication of quantum dots in monolayer molybdenum disulfide (MoS2), and quantum dot arrays with nanometer-scale spatial density by focused electron beam irradiation induced local 2H to 1T phase change in MoS2. By designing the quantum dots in a 2D superlattice, we show that new energy bands form where the new band gap can be controlled by the size and pitch of the quantum dots in the superlattice. The band gap can be tuned from 1.81 eV to 1.42 eV without loss of its photoluminescence performance, which provides new directions for fabricating lasers with designed wavelengths. Our work constitutes a photoresist-free, top-down method to create large-area quantum dot arrays with nanometer-scale spatial density that allow the quantum dots to interfere with each other and create artificial crystals. This technique opens up new pathways for fabricating light emitting devices with 2D materials at desired wavelengths. This demonstration can also enable the assembly of large scale quantum information systems and open up new avenues for the design of artificial 2D materials.

  15. Remedial action plan and site design for stabilization of the inactive uranium mill tailings sites at Slick Rock, Colorado: Remedial Action Selection Report. Preliminary final

    SciTech Connect

    Not Available

    1994-03-01

    This proposed remedial action plan incorporates the results of detailed investigation of geologic, geomorphic, and seismic conditions at the proposed disposal site. The proposed remedial action will consist of relocating the uranium mill tailings, contaminated vicinity property materials, demolition debris, and windblown/waterborne materials to a permanent repository at the proposed Burro Canyon disposal cell. The proposed disposal site will be geomorphically stable. Seismic design parameters were developed for the geotechnical analyses of the proposed cell. Cell stability was analyzed to ensure long-term performance of the disposal cell in meeting design standards, including slope stability, settlement, and liquefaction potential. The proposed cell cover and erosion protection features were also analyzed and designed to protect the RRM (residual radioactive materials) against surface water and wind erosion. The location of the proposed cell precludes the need for permanent drainage or interceptor ditches. Rock to be used on the cell top-, side-, and toeslopes was sized to withstand probable maximum precipitation events.

  16. Catalysis by Design: Well-Defined Single-Site Heterogeneous Catalysts.

    PubMed

    Pelletier, Jérémie D A; Basset, Jean-Marie

    2016-04-19

    Heterogeneous catalysis, a field important industrially and scientifically, is increasingly seeking and refining strategies to render itself more predictable. The main issue is due to the nature and the population of catalytically active sites. Their number is generally low to very low, their "acid strengths" or " redox properties" are not homogeneous, and the material may display related yet inactive sites on the same material. In many heterogeneous catalysts, the discovery of a structure-activity reationship is at best challenging. One possible solution is to generate single-site catalysts in which most, if not all, of the sites are structurally identical. Within this context and using the right tools, the catalyst structure can be designed and well-defined, to reach a molecular understanding. It is then feasible to understand the structure-activity relationship and to develop predictable heterogeneous catalysis. Single-site well-defined heterogeneous catalysts can be prepared using concepts and tools of surface organometallic chemistry (SOMC). This approach operates by reacting organometallic compounds with surfaces of highly divided oxides (or of metal nanoparticles). This strategy has a solid track record to reveal structure-activity relationship to the extent that it is becoming now quite predictable. Almost all elements of the periodical table have been grafted on surfaces of oxides (from simple oxides such as silica or alumina to more sophisticated materials regarding composition or porosity). Considering catalytic hydrocarbon transformations, heterogeneous catalysis outcome may now be predicted based on existing mechanistic proposals and the rules of molecular chemistry (organometallic, organic) associated with some concepts of surface sciences. A thorough characterization of the grafted metal centers must be carried out using tools spanning from molecular organometallic or surface chemistry. By selection of the metal, its ligand set, and the support taken

  17. Molecular docking sites designed for the generation of highly crystalline covalent organic frameworks

    NASA Astrophysics Data System (ADS)

    Ascherl, Laura; Sick, Torben; Margraf, Johannes T.; Lapidus, Saul H.; Calik, Mona; Hettstedt, Christina; Karaghiosoff, Konstantin; Döblinger, Markus; Clark, Timothy; Chapman, Karena W.; Auras, Florian; Bein, Thomas

    2016-04-01

    Covalent organic frameworks (COFs) formed by connecting multidentate organic building blocks through covalent bonds provide a platform for designing multifunctional porous materials with atomic precision. As they are promising materials for applications in optoelectronics, they would benefit from a maximum degree of long-range order within the framework, which has remained a major challenge. We have developed a synthetic concept to allow consecutive COF sheets to lock in position during crystal growth, and thus minimize the occurrence of stacking faults and dislocations. Hereby, the three-dimensional conformation of propeller-shaped molecular building units was used to generate well-defined periodic docking sites, which guided the attachment of successive building blocks that, in turn, promoted long-range order during COF formation. This approach enables us to achieve a very high crystallinity for a series of COFs that comprise tri- and tetradentate central building blocks. We expect this strategy to be transferable to a broad range of customized COFs.

  18. Readability of Online Patient Educational Resources Found on NCI-Designated Cancer Center Web Sites.

    PubMed

    Rosenberg, Stephen A; Francis, David; Hullett, Craig R; Morris, Zachary S; Fisher, Michael M; Brower, Jeffrey V; Bradley, Kristin A; Anderson, Bethany M; Bassetti, Michael F; Kimple, Randall J

    2016-06-01

    The NIH and Department of Health & Human Services recommend online patient information (OPI) be written at a sixth grade level. We used a panel of readability analyses to assess OPI from NCI-Designated Cancer Center (NCIDCC) Web sites. Cancer.gov was used to identify 68 NCIDCC Web sites from which we collected both general OPI and OPI specific to breast, prostate, lung, and colon cancers. This text was analyzed by 10 commonly used readability tests: the New Dale-Chall Readability Formula, Flesch Reading Ease scale, Flesch-Kinaid Grade Level, FORCAST scale, Fry Readability Graph, Simple Measure of Gobbledygook test, Gunning Frequency of Gobbledygook index, New Fog Count, Raygor Readability Estimate Graph, and Coleman-Liau Index. We tested the hypothesis that the readability of NCIDCC OPI was written at the sixth grade level. Secondary analyses were performed to compare readability of OPI between comprehensive and noncomprehensive centers, by region, and to OPI produced by the American Cancer Society (ACS). A mean of 30,507 words from 40 comprehensive and 18 noncomprehensive NCIDCCs was analyzed (7 nonclinical and 3 without appropriate OPI were excluded). Using a composite grade level score, the mean readability score of 12.46 (ie, college level: 95% CI, 12.13-12.79) was significantly greater than the target grade level of 6 (middle-school: P<.001). No difference between comprehensive and noncomprehensive centers was identified. Regional differences were identified in 4 of the 10 readability metrics (P<.05). ACS OPI provides easier language, at the seventh to ninth grade level, across all tests (P<.01). OPI from NCIDCC Web sites is more complex than recommended for the average patient. Copyright © 2016 by the National Comprehensive Cancer Network.

  19. Berm design to reduce risks of catastrophic slope failures at solid waste disposal sites.

    PubMed

    De Stefano, Matteo; Gharabaghi, Bahram; Clemmer, Ryan; Jahanfar, M Ali

    2016-11-01

    Existing waste disposal sites are being strained by exceeding their volumetric capacities because of exponentially increasing rates of municipal solid waste generation worldwide, especially in densely populated metropolises. Over the past 40 years, six well-documented and analyzed disposal sites experienced catastrophic failure. This research presents a novel analysis and design method for implementation of a series of in-situ earth berms to slow down the movement of waste material flow following a catastrophic failure. This is the first study of its kind that employs a dynamic landslide analysis model, DAN-W, and the Voellmy rheological model to approximate solid waste avalanche flow. A variety of single and multiple berm configuration scenarios were developed and tested to find an optimum configuration of the various earth berm geometries and number of berms to achieve desired energy dissipation and reduction in total waste material runout length. The case study application of the novel mitigation measure shows that by constructing a series of six relatively inexpensive 3 m high earth berms at an optimum distance of 250 m from the slope toe, the total runout length of 1000 m and associated fatalities of the Leuwigajah dumpsite catastrophic failure in Bandung, Indonesia, could have been reduced by half. © The Author(s) 2016.

  20. Fly ash design manual for road and site applications. Volume 2: Slurried placement

    SciTech Connect

    McLaren, R.J.; Balsamo, N.J.

    1986-10-01

    This report was produced as part of EPRI Research Project 2422. The goal of this project is to significantly increase the use of fly ash in all high volume uses and to increase its use in medium and high technology applications as well. This report is a utilization manual describing the design of various types of projects where fly ash has been used as a substitute for other common materials in high volume uses. High volume uses of fly ash are typified by structural and non-structural fills for site development, embankments for highways and railroads, backfills, highway subgrade improvement projects and base courses, grouting of subsurface voids, and other similar applications. This report is an integral part of the overall project and has been developed as a design manual pertaining to the use of fly ash as a construction material in these high volume uses. The manual is divided into two volumes. Volume 1 describes the use of fly ash conditioned with small amounts of water to produce a solid, soil-like material. This volume describes the use of fly ash mixed with sufficient water to create a fluid or semi-fluid material. In many applications, Portland cement and aggregate filler are also added to the mixture. The flowable nature of these materials determines the manner in which they are handled, placed, and used in construction. The following uses are discussed: Slurry Backfills, Grouting, and Hydraulic Fills. Miscellaneous uses are also discussed, including the use of fly ash in high fly ash concrete, unsintered synthetic aggregate, and slurry walls. Also included are three appendices containing additional information of interest to the designer: Mix Design, Laboratory Testing, and Bibliography. 19 refs., 53 figs., 12 tabs.

  1. Remedial action plan and site conceptual design for stabilization of the inactive uranium mill tailings site at Ambrosia Lake, New Mexico

    SciTech Connect

    Matthews, M.L. . Uranium Mill Tailings Remedial Action Project Office); Mitzelfelt, R. . Environmental Improvement Div.)

    1991-11-01

    This Remedial Action Plan (RAP) has been developed to serve a dual purpose. It presents the series of activities that is proposed by the US Department of Energy (DOE) to stabilize and control radioactive materials at the inactive Phillips/United Nuclear uranium processing site designated as the Ambrosia Lake site in McKinley County, New Mexico. It also serves to document the concurrence of both State of New Mexico and the US Nuclear Regulatory Commission (NRC) in the remedial action. This agreement, upon execution by the DOE and the state and concurrence by NRC, becomes Appendix B of the Cooperative Agreement.

  2. SDM-Assist software to design site-directed mutagenesis primers introducing “silent” restriction sites

    PubMed Central

    2013-01-01

    Background Over the past decades site-directed mutagenesis (SDM) has become an indispensable tool for biological structure-function studies. In principle, SDM uses modified primer pairs in a PCR reaction to introduce a mutation in a cDNA insert. DpnI digestion of the reaction mixture is used to eliminate template copies before amplification in E. coli; however, this process is inefficient resulting in un-mutated clones which can only be distinguished from mutant clones by sequencing. Results We have developed a program – ‘SDM-Assist’ which creates SDM primers adding a specific identifier: through additional silent mutations a restriction site is included or a previous one removed which allows for highly efficient identification of ‘mutated clones’ by a simple restriction digest. Conclusions The direct identification of SDM clones will save time and money for researchers. SDM-Assist also scores the primers based on factors such as Tm, GC content and secondary structure allowing for simplified selection of optimal primer pairs. PMID:23522286

  3. NMR studies of abasic sites in DNA duplexes: deoxyadenosine stacks into the helix opposite the cyclic analog of 2-deoxyribose

    SciTech Connect

    Kalnik, M.W.; Chang, C.N.; Grollman, A.P.; Patel, D.J.

    1988-02-09

    Proton and phosphorus NMR studies are reported for the complementary d(C-A-T-G-A-G-T-A-C) x d(G-T-A-C-F-C-A-T-G) nonanucleotide duplex (designated AP/sub F/ 9-mer duplex) which contains a stable abasic site analog, F, in the center of the helix. This oligodeoxynucleotide contains a modified tetrahydrofuran moiety, isosteric with 2-deoxyribofuranose, which serves as a structural analog of a natural apurinic/apyrimidinic site. Exchangeable and nonexchangeable base and sugar protons, including those located at the abasic site, have been assigned in the complementary AP/sub F/ 9-mer duplex by recording and analyzing two-dimensional phase-sensitive NOESY data sets in H/sub 2/O and D/sub 2/O solution at low temperature (0/sup 0/C). These studies indicate that A5 inserts into the helix opposite the abasic site F14 and stacks with flanking G4 x C15 and G6 x C13 Watson-Crick base pairs. Base-sugar proton NOE connectivities were measured through G4-A5-G6 on the unmodified strand and between the base protons of C15 and the sugar protons of the 5'-flanking residue F14 on the modified strand. These studies establish that all glycosidic torsion angles are anti and that the helix is right-handed at and adjacent to the abasic site in the AP/sub F/ 9-mer duplex. Two of the 16 phosphodiester groups exhibit phosphorus resonances outside the normal spectral dispersion indicative of altered torsion angles at two of the phosphate groups in the backbone of the AP/sub F/ 9-mer duplex.

  4. Activation of GLP-1 Receptor Enhances Neuronal Base Excision Repair via PI3K-AKT-Induced Expression of Apurinic/Apyrimidinic Endonuclease 1

    PubMed Central

    Yang, Jenq-Lin; Chen, Wei-Yu; Chen, Yin-Ping; Kuo, Chao-Ying; Chen, Shang-Der

    2016-01-01

    Glucagon-like peptide-1 (GLP-1) is an intestinal-secreted incretin that increases cellular glucose up-take to decrease blood sugar. Recent studies, however, suggest that the function of GLP-1 is not only to decrease blood sugar, but also acts as a neurotrophic factor that plays a role in neuronal survival, neurite outgrowth, and protects synaptic plasticity and memory formation from effects of β-amyloid. Oxidative DNA damage occurs during normal neuron-activity and in many neurological diseases. Our study describes how GLP-1 affected the ability of neurons to ameliorate oxidative DNA damage. We show that activation of GLP-1 receptor (GLP-1R) protect cortical neurons from menadione induced oxidative DNA damage via a signaling pathway involving enhanced DNA repair. GLP-1 stimulates DNA repair by activating the cyclic AMP response element binding protein (CREB) which, consequently, induces the expression of apurinic/apyrimidinic endonuclease 1 (APE1), a key enzyme in the base excision DNA repair (BER) pathway. In this study, APE1 expression was down-regulated as a consequence phosphatidylinositol-3 kinase (PI3K) suppression by the inhibitor LY294002, but not by the suppression of MEK activity. Ischemic stroke is typically caused by overwhelming oxidative-stress in brain cells. Administration of exentin-4, an analogue of GLP-1, efficiently enhanced DNA repair in brain cells of ischemic stroke rats. Our study suggests that a new function of GLP-1 is to elevate DNA repair by inducing the expression of the DNA repair protein APE1. PMID:27698937

  5. 17β-estradiol increases expression of the oxidative stress response and DNA repair protein apurinic endonuclease (Ape1) in the cerebral cortex of female mice following hypoxia.

    PubMed

    Dietrich, Alicia K; Humphreys, Gwendolyn I; Nardulli, Ann M

    2013-11-01

    While it is well established that 17β-estradiol (E2) protects the rodent brain from ischemia-induced damage, it has been unclear how this neuroprotective effect is mediated. Interestingly, convincing evidence has also demonstrated that maintaining or increasing the expression of the oxidative stress response and DNA repair protein apurinic endonuclease 1 (Ape1) is instrumental in reducing ischemia-induced damage in the brain. Since E2 increases expression of the oxidative stress response proteins Cu/Zn superoxide dismutase and thioredoxin in the brain, we hypothesized that E2 may also increase Ape1 expression and that this E2-induced expression of Ape1 may help to mediate the neuroprotective effects of E2 in the brain. To test this hypothesis, we utilized three model systems including primary cortical neurons, brain slice cultures, and whole animals. Although estrogen receptor α and Ape1 were expressed in primary cortical neurons, E2 did not alter Ape1 expression in these cells. However, immunofluorescent staining and quantitative Western blot analysis demonstrated that estrogen receptor α and Ape1 were expressed in the nuclei of cortical neurons in brain slice cultures and that E2 increased Ape1 expression in the cerebral cortex of these cultures. Furthermore, Ape1 expression was increased and oxidative DNA damage was decreased in the cerebral cortices of ovariectomized female C57Bl/6J mice that had been treated with E2 and exposed to hypoxia. Taken together, our studies demonstrate that the neuronal microenvironment may be required for increased Ape1 expression and that E2 enhances expression of Ape1 and reduces oxidative DNA damage, which may in turn help to reduce ischemia-induced damage in the cerebral cortex and mediate the neuroprotective effects of E2.

  6. Copper-zinc superoxide dismutase prevents the early decrease of apurinic/apyrimidinic endonuclease and subsequent DNA fragmentation after transient focal cerebral ischemia in mice.

    PubMed

    Fujimura, M; Morita-Fujimura, Y; Narasimhan, P; Copin, J C; Kawase, M; Chan, P H

    1999-11-01

    DNA damage and its repair mechanism are thought to be involved in ischemia/reperfusion injury in the brain. We have previously shown that apurinic/apyrimidinic endonuclease (APE/Ref-1), a multifunctional protein in the DNA base excision repair pathway, rapidly decreased after transient focal cerebral ischemia (FCI) before the peak of DNA fragmentation. To further investigate the role of reactive oxygen species in APE/Ref-1 expression in vivo, we examined the expression of APE/Ref-1 and DNA damage after FCI in wild-type and transgenic mice overexpressing copper-zinc superoxide dismutase. Transgenic mice overexpressing copper-zinc superoxide dismutase and wild-type littermates were subjected to 60 minutes of transient FCI by intraluminal blockade of the middle cerebral artery. APE/Ref-1 protein expression was analyzed by immunohistochemistry and Western blot analysis. DNA damage was evaluated by gel electrophoresis and terminal deoxynucleotidyl transferase-mediated uridine 5'-triphosphate-biotin nick end-labeling (TUNEL). A similar level of APE/Ref-1 was detected in the control brains from both groups. APE/Ref-1 was significantly reduced 1 hour after transient FCI in both groups, whereas the transgenic mice had less reduction than that seen in wild-type mice 1 and 4 hours after FCI. DNA laddering was detected 24 hours after FCI and was decreased in transgenic mice. Double staining with APE/Ref-1 and TUNEL showed that the neurons that lost APE/Ref-1 immunoreactivity became TUNEL positive. These results suggest that reactive oxygen species contribute to the early decrease of APE/Ref-1 and thereby exacerbate DNA fragmentation after transient FCI in mice.

  7. 76 FR 43685 - Designation of an Ocean Dredged Material Disposal Site (ODMDS) in the Gulf of Mexico Off the...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-21

    ... AGENCY Designation of an Ocean Dredged Material Disposal Site (ODMDS) in the Gulf of Mexico Off the Mouth... of an ODMDS in the Gulf of Mexico off the mouth of the Atchafalaya River, St. Mary Parish, LA... the designation of an ODMDS in the Gulf of Mexico off the mouth of the Atchafalaya River, St. Mary...

  8. 77 FR 67399 - Hewlett-Packard Company, Design Delivery Organization (DDO), Including On-Site Leased Workers...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-11-09

    ... On-Site Leased Workers From Manpower, Synova Inc., and Pinnacle Technical Resources, Corvallis, OR; Hewlett-Packard Company, Ink Jet & Web Services, World Wide Design Group, Vancouver, WA; Amended..., Ink Jet & Web Services, World Wide Design Group, Vancouver, Washington ] operated in conjunction...

  9. Use of an Improved Matching Algorithm to Select Scaffolds for Enzyme Design Based on a Complex Active Site Model

    PubMed Central

    Huang, Xiaoqiang; Xue, Jing; Lin, Min; Zhu, Yushan

    2016-01-01

    Active site preorganization helps native enzymes electrostatically stabilize the transition state better than the ground state for their primary substrates and achieve significant rate enhancement. In this report, we hypothesize that a complex active site model for active site preorganization modeling should help to create preorganized active site design and afford higher starting activities towards target reactions. Our matching algorithm ProdaMatch was improved by invoking effective pruning strategies and the native active sites for ten scaffolds in a benchmark test set were reproduced. The root-mean squared deviations between the matched transition states and those in the crystal structures were < 1.0 Å for the ten scaffolds, and the repacking calculation results showed that 91% of the hydrogen bonds within the active sites are recovered, indicating that the active sites can be preorganized based on the predicted positions of transition states. The application of the complex active site model for de novo enzyme design was evaluated by scaffold selection using a classic catalytic triad motif for the hydrolysis of p-nitrophenyl acetate. Eighty scaffolds were identified from a scaffold library with 1,491 proteins and four scaffolds were native esterase. Furthermore, enzyme design for complicated substrates was investigated for the hydrolysis of cephalexin using scaffold selection based on two different catalytic motifs. Only three scaffolds were identified from the scaffold library by virtue of the classic catalytic triad-based motif. In contrast, 40 scaffolds were identified using a more flexible, but still preorganized catalytic motif, where one scaffold corresponded to the α-amino acid ester hydrolase that catalyzes the hydrolysis and synthesis of cephalexin. Thus, the complex active site modeling approach for de novo enzyme design with the aid of the improved ProdaMatch program is a promising approach for the creation of active sites with high catalytic

  10. Use of an Improved Matching Algorithm to Select Scaffolds for Enzyme Design Based on a Complex Active Site Model.

    PubMed

    Huang, Xiaoqiang; Xue, Jing; Lin, Min; Zhu, Yushan

    2016-01-01

    Active site preorganization helps native enzymes electrostatically stabilize the transition state better than the ground state for their primary substrates and achieve significant rate enhancement. In this report, we hypothesize that a complex active site model for active site preorganization modeling should help to create preorganized active site design and afford higher starting activities towards target reactions. Our matching algorithm ProdaMatch was improved by invoking effective pruning strategies and the native active sites for ten scaffolds in a benchmark test set were reproduced. The root-mean squared deviations between the matched transition states and those in the crystal structures were < 1.0 Å for the ten scaffolds, and the repacking calculation results showed that 91% of the hydrogen bonds within the active sites are recovered, indicating that the active sites can be preorganized based on the predicted positions of transition states. The application of the complex active site model for de novo enzyme design was evaluated by scaffold selection using a classic catalytic triad motif for the hydrolysis of p-nitrophenyl acetate. Eighty scaffolds were identified from a scaffold library with 1,491 proteins and four scaffolds were native esterase. Furthermore, enzyme design for complicated substrates was investigated for the hydrolysis of cephalexin using scaffold selection based on two different catalytic motifs. Only three scaffolds were identified from the scaffold library by virtue of the classic catalytic triad-based motif. In contrast, 40 scaffolds were identified using a more flexible, but still preorganized catalytic motif, where one scaffold corresponded to the α-amino acid ester hydrolase that catalyzes the hydrolysis and synthesis of cephalexin. Thus, the complex active site modeling approach for de novo enzyme design with the aid of the improved ProdaMatch program is a promising approach for the creation of active sites with high catalytic

  11. Early Site Permit Demonstration Program: Guidelines for determining design basis ground motions. Volume 1

    SciTech Connect

    Not Available

    1993-03-18

    This report develops and applies a methodology for estimating strong earthquake ground motion. The motivation was to develop a much needed tool for use in developing the seismic requirements for structural designs. An earthquake`s ground motion is a function of the earthquake`s magnitude, and the physical properties of the earth through which the seismic waves travel from the earthquake fault to the site of interest. The emphasis of this study is on ground motion estimation in Eastern North America (east of the Rocky Mountains), with particular emphasis on the Eastern United States and southeastern Canada. Eastern North America is a stable continental region, having sparse earthquake activity with rare occurrences of large earthquakes. While large earthquakes are of interest for assessing seismic hazard, little data exists from the region to empirically quantify their effects. Therefore, empirically based approaches that are used for other regions, such as Western North America, are not appropriate for Eastern North America. Moreover, recent advances in science and technology have now made it possible to combine theoretical and empirical methods to develop new procedures and models for estimating ground motion. The focus of the report is on the attributes of ground motion in Eastern North America that are of interest for the design of facilities such as nuclear power plants. Specifically considered are magnitudes M from 5 to 8, distances from 0 to 500 km, and frequencies from 1 to 35 Hz.

  12. The de novo design of a rubredoxin-like Fe site.

    PubMed

    Farinas, E; Regan, L

    1998-09-01

    A redox center similar to that of rubredoxin was designed into the 56 amino acid immunoglobulin binding B1 domain of Streptococcals protein G. The redox center in rubredoxin contains an iron ion tetrahedrally coordinated by four cysteine residues, [Fe(S-Cys)4](-1),(-2). The design criteria for the target site included taking backbone movements into account, tetrahedral metal-binding, and maintaining the structure and stability of the wild-type protein. The optical absorption spectrum of the Co(II) complex of the metal-binding variant is characteristic of tetrahedral chelation by four cysteine residues. Circular dichroism and nuclear magnetic resonance measurements reveal that the metal-free and Cd(II)-bound forms of the variant are folded correctly and are stable. The Fe(III) complex of the metal-binding mutant reproduces the optical and the electron paramagnetic resonance spectra of oxidized rubredoxin. This demonstrates that the engineered protein chelates Fe(III) in a tetrahedral array, and the resulting center is similar to that of oxidized rubredoxin.

  13. Remedial Action Plan and site design for stabilization of the inactive uranium mill tailings site at Mexican Hat, Utah: Appendix E. Final report

    SciTech Connect

    1988-07-01

    This document provides Appendix E of the Remedial Action Plan (RAP) presented in 1988 for the stabilization of the inactive uranium mill tailings at the Mexican Hat, Utah site. The RAP was developed to serve a two- fold purpose. It presents the activities proposed by the Department of Energy (DOE) to accomplish long-term stabilization and control of the residual radioactive materials (RRM) from Monument Valley, Arizona, and Mexican Hat, Utah, at the Mexican Hat disposal site. It also serves to document the concurrence of both the Navajo Nation and the Nuclear Regulatory Commission (NRC) in the remedial action. This agreement, upon execution by the DOE and the Navajo Nation and concurrence by the NRC, becomes Appendix B of the Cooperative Agreement. The RAP has been structured to provide a comprehensive understanding of the remedial action proposed for the Monument Valley and Mexican Hat sites. It includes specific design and construction requirements for the remedial action.

  14. Remedial Action Plan and site design for stabilization of the inactive uranium mill tailings site at Rifle, Colorado. Volume 1, Text: Appendices A, B, and C: Final report

    SciTech Connect

    1990-02-01

    This document has been structured to provide a comprehensive understanding of the remedial action proposed for the Rifle sites. That remedial action consists of removing approximately 4,185,000 cubic yards (cy) of tailings and contaminated materials from their current locations, transporting, and stabilizing the tailings material at the Estes Gulch disposal site, approximately six miles north of Rifle. The tailings and contaminated materials are comprised of approximately 597,000 cy from Old Rifle, 3,232,000 cy from New Rifle, and 322,000 cy from vicinity properties and about 34,000 cy from demolition. The remedial action plan includes specific design requirements for the detailed design and construction of the remedial action. An extensive amount of data and supporting information have been generated for this remedial action and cannot all be incorporated into this document. Pertinent information and data are included with reference given to the supporting documents.

  15. Design-Basis Flood Estimation for Site Characterization at Nuclear Power Plants in the United States of America

    SciTech Connect

    Prasad, Rajiv; Hibler, Lyle F.; Coleman, Andre M.; Ward, Duane L.

    2011-11-01

    The purpose of this document is to describe approaches and methods for estimation of the design-basis flood at nuclear power plant sites. Chapter 1 defines the design-basis flood and lists the U.S. Nuclear Regulatory Commission's (NRC) regulations that require estimation of the design-basis flood. For comparison, the design-basis flood estimation methods used by other Federal agencies are also described. A brief discussion of the recommendations of the International Atomic Energy Agency for estimation of the design-basis floods in its member States is also included.

  16. Rational Catalyst Design of Titanium-Silica Materials Aided by Site-Specific Titration Tools

    NASA Astrophysics Data System (ADS)

    Eaton, Todd Robert

    between particle and crystal size, as obtained from XRD. In the course of establishing these relationships we've gained the knowledge of how to control TiO x structure, which enables the design of new and better catalysts. Understanding the synthesis-structure-function relationships allow for the design of a tandem photo/thermocatalytic reaction system for producing and consuming H2O2. By partially overcoating a TiO 2 photocatalyst with a ˜2 nm silica layer we observe a 56-fold rate improvement compared to bare-TiO2 for H2O2 synthesis from the proton-assisted reduction of O2. Addition of metal-SiO2 thermocatalysts (metal=Ti, Nb, or Ta) with sites needed for H2O2 activation creates a tandem system wherein the H2O2 produced in situ is utilized for alkene epoxidation. Compared to a thermocatalytic-only system, the tandem system accelerates epoxidation for cis-cyclooctene(11x faster), styrene(20x) and 1-octene(30x). This approach demonstrates a means for epoxidation with O2 that avoids H2O2 purification and transport, simplifies the total process, provides new opportunities for control by independent H2O2 production and consumption in the same reactor, and enhances rates relative to thermocatalytic-only epoxidation by intimately coupling H2O2 generation and consumption. Critically, establishment of titanium-silica synthesis-structure-function relationships enables the design of new catalysts and systems that are less energy- and material-intensive, leading towards more sustainable chemistry.

  17. Seismic design of circular-section concrete-lined underground openings: Preclosure performance considerations for the Yucca Mountain Site

    SciTech Connect

    Richardson, A.M.; Blejwas, T.E.

    1992-07-01

    Yucca Mountain, the potential site of a repository for high-level radioactive waste, is situated in a region of natural and man-made seismicity. Underground openings excavated at this site must be designed for worker safety in the seismic environment anticipated for the preclosure period. This includes accesses developed for site characterization regardless of the ultimate outcome of the repository siting process. Experience with both civil and mining structures has shown that underground openings are much more resistant to seismic effects than surface structures, and that even severe dynamic strains can usually be accommodated with proper design. This paper discusses the design and performance of lined openings in the seismic environment of the potential site. The types and ranges of possible ground motions (seismic loads) are briefly discussed. Relevant historical records of underground opening performance during seismic loading are reviewed. Simple analytical methods of predicting liner performance under combined in situ, thermal, and seismic loading are presented, and results of calculations are discussed in the context of realistic performance requirements for concrete-lined openings for the preclosure period. Design features that will enhance liner stability and mitigate the impact of the potential seismic load are reviewed. The paper is limited to preclosure performance concerns involving worker safety because present decommissioning plans specify maintaining the option for liner removal at seal locations, thus decoupling liner design from repository postclosure performance issues.

  18. Remedial action plan and site design for stabilization of the inactive uranium mill tailings site at Lakeview, Oregon: Volume 1, Text and appendices A through D

    SciTech Connect

    Chernoff, A R; Ervin, C

    1992-07-01

    The Lakeview inactive uranium processing site is in Lake County, Oregon, approximately one mile northwest of the town of Lakeview, sixteen miles north of the California-Oregon border, and 96 miles east of Klamath Falls. The total designated site covers an area of 258 acres consisting of a tailings pile (30 acres). seven evaporation ponds (69 acres), the mill buildings, and related structures. The mill buildings and other structures have been decontaminated and are currently being used by Goose Lake Lumber Company. The tailings pile at the processing site was originally stabilized by Atlantic Richfield with an earthen cover 18--24 inches thick. The average depth of the tailings, including the cover, varied from six to eight feet. There were estimated to be 662,000 cubic yards of tailings, windblown contaminated materials, and vicinity property materials. During remedial action under the Uranium Mill Tailings Remedial Action (UMTRA) Project, approximately 264,000 cubic yards of additional contaminated materials were identified from excavations required to remove thorium- and arsenic-contaminated soils. The remedial action for the Lakeview site consisted of the cleanup, relocation, consolidation, and stabilization of all residual radioactive materials and thorium- and arsenic-contaminated materials in a partially below-grade disposal cell at a location approximately seven miles northwest of the tailings site, identified as the Collins Ranch site. A cover, including a radon/infiltration barrier and rock layer for protection from erosion, was Placed on top of the tailings. A rock-soil matrix covers the topslope and provides a growth medium for vegetation. The US Department of Energy (DOE) will retain the license and surveillance and maintenance responsibilities for the final restricted site of 13 acres.

  19. Pilot-scale demonstration of surfactant-enhanced PCE solubilization at the Bachman Road site. 1. Site characterization and test design.

    PubMed

    Abriola, Linda M; Drummond, Chad D; Hahn, Ernest J; Hayes, Kim F; Kibbey, Tohren C G; Lemke, Lawrence D; Pennell, Kurt D; Petrovskis, Erik A; Ramsburg, C Andrew; Rathfelder, Klaus M

    2005-03-15

    A pilot-scale demonstration of surfactant-enhanced aquifer remediation (SEAR) was conducted to recover dense nonaqueous phase liquid (DNAPL) tetrachloroethene (PCE) from a sandy glacial outwash aquifer underlying a former dry cleaning facility at the Bachman Road site in Oscoda, MI. Part one of this two-part paper describes site characterization efforts and a comprehensive approach to SEAR test design, effectively integrating laboratory and modeling studies. Aquifer coring and drive point sampling suggested the presence of PCE-DNAPL in a zone beneath an occupied building. A narrow PCE plume emanating from the vicinity of this building discharges into Lake Huron. The shallow unconfined aquifer, characterized by relatively homogeneous fine-medium sand deposits, an underlying clay layer, and the absence of significant PCE transformation products, was judged suitable for the demonstration of SEAR. Tween 80 was selected for application based upon its favorable solubilization performance in batch and two-dimensional sand tank treatability studies, biodegradation potential, and regulatory acceptance. Three-dimensional flow and transport models were employed to develop a robust design for surfactant delivery and recovery. Physical and fiscal constraints led to an unusual hydraulic design, in which surfactant was flushed across the regional groundwater gradient, facilitating the delivery of concentrations of Tween 80 exceeding 1% (wt) throughout the treatment zone. The potential influence of small-scale heterogeneity on PCE-DNAPL distribution and SEAR performance was assessed through numerical simulations incorporating geostatistical permeability fields based upon available core data. For the examined conditions simulated PCE recoveries ranged from 94to 99%. The effluent treatment system design consisted of low-profile air strippers coupled with carbon adsorption to trap off-gas PCE and discharge of treated aqueous effluent to a local wastewater treatment plant. The

  20. Wind load comparison for the ASCE standard 7 and the Hanford site design criteria

    SciTech Connect

    Giller, R.A., Westinghouse Hanford

    1996-07-16

    This document provides calculations and discussions to compare Hanford Site wind load criteria with the current national standard for wind loads (ASCE 7, 1995). Site criteria uses the 1988 edition ASCE 7.

  1. Paleolimnology Web Portal: A Web Site Designed to Increase Paleolimnology Data Availability

    NASA Astrophysics Data System (ADS)

    Eakin, C. M.; Moy, C. M.; Habermann, T.; Gross, W. S.; Keltner, J. M.

    2001-12-01

    Despite widespread use of lacustrine records to interpret paleolimnologic and paleoclimatic change, there is a large gap between the data published in peer-reviewed journals and those submitted for archive and available to other researchers online. A primary goal of the World Data Center (WDC) for paleoclimatology and the International Geosphere-Biosphere Programme (IGBP) - Past Global Changes (PAGES) core programme is to have full and open sharing of all data sets needed for global change studies. To help improve the quantity and quality of data submitted to the WDC for Paleoclimatology, we are developing online data submission and advanced interactive browse and access tools. Our poster presents a new web-site designed to make paleolimnology data more accessible by incorporating web-based data submission forms, a multi-proxy relational database, and interactive mapping tools. The WDC for Paleoclimatology is currently designing intuitive and streamlined web-based submission forms, which will allow investigators to quickly submit their data and metadata on-line. We are also importing all existing data and metadata in our archives into a multiproxy relational database that will allow users to quickly query and retrieve paleolimnological data, as well as display the data in various formats. Furthermore, we are implementing two Paleolimnology mapping tools that will allow users to search, display, and query data in a geographical format. The first tool, WebMapper, uses a Java applet to draw maps and display metadata. This will be supplemented by a plotting tool that will provide basic plotting functions to allow users to examine data before downloading them. The second mapping tool, ArcIMS, allows users to overlay paleoclimatic data with various GIS data sets in addition to providing basic spatial analysis functions. We believe that these new web-based features will encourage more extensive data sharing and submission, making paleolimnological data more available and

  2. Site study plan for Exploratory shaft facilities design foundation boreholes (shaft surface facility foundation borings), Deaf Smith County Site, Texas: Surface-based geotechnical field program: Preliminary draft

    SciTech Connect

    Not Available

    1987-12-01

    This site study plan describes the Exploratory Shaft Facilities (ESF) Design Foundation Boreholes field activities to be conducted during early stages of Site Characterization at the Deaf Smith County, Texas, site. The field program has been designed to provide data useful in addressing information/data needs resulting from federal/state/local regulations, and repository program requirements. Approximately 50 foundation boreholes will be drilled within the ESP location to provide data necessary for design of the ESF and to satisfy applicable shaft permitting requirements. Soils and subsurface rock will be sampled as the foundation boreholes are advanced. Soil samples or rock core will be taken through the Blackwater Draw and Ogallala Formations and the Dockum Group. Hydrologic testing will be performed in boreholes that penetrates the water table. In-situ elastic properties will be determined from both the soil strata and rock units along the length of the boreholes. Field methods/tests are chosen that provide the best or only means of obtaining the required data. The Salt Repository Project (SRP) Networks specify the schedule under which the program will operate. Drilling will not begin until after site ground water baseline conditions have been established. The Technical Field Services Contractor is responsible for conducting the field program of drilling and testing. Samples and data will be handled and reported in accordance with established SRP procedures. A quality assurance program will be utilized to assure that activities affecting quality are performed correctly and that the appropriate documentation is maintained. 25 refs., 10 figs., 6 tabs.

  3. Site-specific integration and tailoring of cassette design for sustainable gene transfer.

    PubMed

    Lombardo, Angelo; Cesana, Daniela; Genovese, Pietro; Di Stefano, Bruno; Provasi, Elena; Colombo, Daniele F; Neri, Margherita; Magnani, Zulma; Cantore, Alessio; Lo Riso, Pietro; Damo, Martina; Pello, Oscar M; Holmes, Michael C; Gregory, Philip D; Gritti, Angela; Broccoli, Vania; Bonini, Chiara; Naldini, Luigi

    2011-08-21

    Integrative gene transfer methods are limited by variable transgene expression and by the consequences of random insertional mutagenesis that confound interpretation in gene-function studies and may cause adverse events in gene therapy. Site-specific integration may overcome these hurdles. Toward this goal, we studied the transcriptional and epigenetic impact of different transgene expression cassettes, targeted by engineered zinc-finger nucleases to the CCR5 and AAVS1 genomic loci of human cells. Analyses performed before and after integration defined features of the locus and cassette design that together allow robust transgene expression without detectable transcriptional perturbation of the targeted locus and its flanking genes in many cell types, including primary human lymphocytes. We thus provide a framework for sustainable gene transfer in AAVS1 that can be used for dependable genetic manipulation, neutral marking of the cell and improved safety of therapeutic applications, and demonstrate its feasibility by rapidly generating human lymphocytes and stem cells carrying targeted and benign transgene insertions.

  4. Chemical reactivity at an antibody binding site elicited by mechanistic design of a synthetic antigen.

    PubMed Central

    Tramontano, A; Janda, K D; Lerner, R A

    1986-01-01

    Monoaryl phosphonate esters, designated as analogs of the transition state in the hydrolysis of carboxylic esters, were synthesized and used as haptens to generate specific monoclonal antibodies. Some of these antibodies react with cognate aryl carboxylic esters to release a fluorescent alcohol. The reaction appears to be stoichiometric; however, the activity is slowly regenerated under alkaline conditions or by treatment with hydroxylamine. Specificity is rigorous for esters of p-trifluoroacetamidophenylacetic acid, demonstrating a structural correspondence with the phosphonate hapten. Saturation kinetics are observed and kinetic parameters (kmax, Vmax, and Km) are reported. The haptenic phosphonate is a competitive inhibitor of the reaction (Ki, 35 nM); whereas the carboxylate product of ester hydrolysis is a less effective inhibitor (Ki, ca. 7500 nM). Chemical modification of side chain groups in the protein show a partial reduction in activity on acylation of lysine or nitration of tyrosine and a dramatic quenching upon modification of histidine. The evidence is discussed in terms of a mechanism in which amino acids of the antibody combining site participate in nucleophilic and/or general base catalysis. The properties of this system suggest that it is an example of enzymic transacylation where a deacylation step is not catalyzed. The possibility of deriving enzymic function from immunological specificity through this approach is advanced. PMID:3462723

  5. Geotechnical, Hydrogeologic and Vegetation Data Package for 200-UW-1 Waste Site Engineered Surface Barrier Design

    SciTech Connect

    Ward, Andy L.

    2007-11-26

    Fluor Hanford (FH) is designing and assessing the performance of engineered barriers for final closure of 200-UW-1 waste sites. Engineered barriers must minimize the intrusion and water, plants and animals into the underlying waste to provide protection for human health and the environment. The Pacific Northwest National Laboratory (PNNL) developed Subsurface Transport Over Multiple Phases (STOMP) simulator is being used to optimize the performance of candidate barriers. Simulating barrier performance involves computation of mass and energy transfer within a soil-atmosphere-vegetation continuum and requires a variety of input parameters, some of which are more readily available than others. Required input includes parameter values for the geotechnical, physical, hydraulic, and thermal properties of the materials comprising the barrier and the structural fill on which it will be constructed as well as parameters to allow simulation of plant effects. This report provides a data package of the required parameters as well as the technical basis, rationale and methodology used to obtain the parameter values.

  6. Designing an in-flight airborne calibration site using experience from vicarious radiometric satellite calibration

    NASA Astrophysics Data System (ADS)

    Livens, Stefan; Debruyn, Walter; Sterckx, Sindy; Reusen, Ils

    2011-11-01

    Laboratory calibration of electro-optical sensors is preferably complemented by regular in-flight verification. This checks whether the lab calibration parameters remain valid or recalibration is necessary. In-flight verification can be achieved by vicarious calibration using in-flight measurements of calibration targets. We intend to identify and design a set of suitable radiometric calibration targets. For this, we borrow from expertise gained with the PROBA-V satellite calibration system, which uses multiple vicarious methods relying on diverse natural on-ground targets. Besides reflectance based calibration using ground measurements, the PROBA-V calibration methods are unproven for use in airborne calibration. The selected targets should be suitable for the calibration of both multispectral and hyperspectral imagers. We start from general requirements for radiometric targets and investigate their applicability to airborne calibration. From this we identify two possible sets of natural calibration sites in Belgium. One set, located in the Campine region, contains small water bodies and sandy lakesides. Another set is located in the Westhoek region near the Belgian coast. It offers better suitable water bodies, as well as sandy areas, grass fields and dark targets. Airborne calibration lends itself to the use of smaller artifical targets. We propose to complement the natural targets with a portable target consisting of agricultural nets with different densities. The definition of sets of calibration targets, both natural and artificial can facilitate the investigation of the usability of vicarious targets and method for inflight radiometric verification.

  7. Preliminary design study of underground pumped hydro and compressed-air energy storage in hard rock. Volume 5: Site selection

    NASA Astrophysics Data System (ADS)

    1981-04-01

    A six-step site selection process undertaken to identify and subsequently rank potential sites suitable for either an underground pumped hydroelectric (UPH) facility, or a water-compensated hard-rock cavern compressed air energy storage (CAES) facility is described. The region of study was confined to the service area of the Potomac Electric Power Company (PEPCO) and contiguous areas. Overriding considerations related to geology, environmental impact and transmission-line routing were studies within the context of minimizing plant costs. The selection process led to the identification of several sites suitable for the development of either a CAES or an UPH facility. Design development and site exploration at the selected site are described.

  8. Subsurface moisture regimes and tracer movement under two types of trench-cap designs for shallow land burial sites

    SciTech Connect

    Perkins, B.A.; Cokal, E.J.

    1986-03-01

    The Los Alamos work has focused on proper design of shallow land burial (SLB) sites in arid and semiarid regions and on applying corrective measures to existing sites. One of the most important design features affecting the probability of movement of radionuclides in SLB sites is the type of trench cap placed over the waste. The cap influences such interdependent parameters as erosion, water infiltration and percolation, and biointrusion. To obtain experimental data for arid and semiarid sites, two different designs of trench caps, one with topsoil underlain with a cobble/gravel biobarrier and one with topsoil underlain with crushed tuff, were compared with respect to (1) seasonal changes in volumetric soil water content, and (2) downward migration of tracers emplaced directly below each type of trench cap. The causes for the large differences in concentrations found in this experiment need to be investigated further. Problems in environmental modeling and monitoring of arid and semiarid SLB sites because of heterogeneities in the soil profiles and their implications for SLB waste management need to be better understood. More work in trench-cap design and its influence on the many pathways available for mobilization is needed.

  9. Site Selection for the European ELT: working package included in the European FP6 ``ELT design study'' contract

    NASA Astrophysics Data System (ADS)

    Muñoz-Tuñón, C.; Sarazin, M.; Vernin, J.

    2007-10-01

    The site selection for the future European Large Telescope (E-ELT) is a key issue within the European proposal funded by the European Union (EU), within the ``ELT design study'' proposal. The organization, working scheme and baseline frameworks are reviewed. For the definition of the working package WP12000 ``Site Characterization'', important use has been done of previous work in the definition of techniques and tools for the study of the atmosphere above observing sites. We have also taken advantage of the number of data already available which have naturally defined a ranking among the known places which have also been taken as a base line for pre-selecting the candidate sites. The work will last 4 years, it started in 2005 and is organized in subtasks, working packages WP, whose main objectives are the following: WP12100: to characterize two top astronomical sites (ORM and North-Paranal) and to explore three other alternatives (Macon in Argentina, Izaña in Spain and Aklim in Morocco) suitable to install an ELT under the best conditions (Dome C is been currently under investigation, and no particular effort will be put in this site, but rather its atmospheric properties will be compared to the above mentioned sites). WP12200 is dedicated to design, build and operate a set of standard equipment in all the sites and to perform long term campaign. WP12300 will investigate wavefront properties over large baselines (50-100 m) corresponding to the size of the future ELT, as well as the fine characterization of the optical turbulence within the boundary layer. A similar plan is being carried out by the Thirty Meter Telescope (TMT) site selection team. For the sake of saving resources (budget and people), the TMT preselected sites (all in the American Continent) are not included in our European study.

  10. Modification No. 2 to the remedial action plan and site design for stabilization of the inactive uranium mill tailings site at Green River, Utah: Final

    SciTech Connect

    1996-11-01

    Portions of the final Remedial Action Plan (RAP) for the Green River site, Volumes 1 and 2, Appendix B of the Cooperative Agreement No. DE-FC04-81AL16257, March 1991 (DOE, 1991) have been modified. The changes to the RAP are designated as RAP Modification No. 2. These changes have been placed in a three-ring binder that will supplement the original RAP (DOE, 1991), and include the following: addendum to the Executive Summary; Section 3.5 (Ground Water part of the Site Characterization Summary); Section 4.0 (Site Design); Section5.0 (Water Resources Protection Strategy Summary); Appendix D.5 (Ground Water Hydrology); and Appendix E (Ground Water Protection Strategy). In addition to these revisions, there have been editorial changes that clarify the text, but do not change the meaning. Also, certain sections of the document, which are included in the submittal for ease of review and continuity, have been updated to reflect the final ground water protection standards and the current UMTRA Project format and content of RAPs.

  11. Remedial Action Plan and site design for stabilization of the inactive uranium mill tailings site at Durango, Colorado: Remedial action selection report

    SciTech Connect

    Not Available

    1991-12-01

    The uranium mill tailings site near Durango, Colorado, was one of 24 inactive uranium mill sites designated to be remediated by the US Department of Energy (DOE) under the Uranium Mill Tailings Radiation Control Act of 1978 (UMTRCA). Part of the UMTRCA requires that the US Nuclear Regulatory Commission (NRC) concur with the DOE's Remedial Action Plan (RAP) and certify that the remedial action conducted at the site complies with the standards promulgated by the US Environmental Protection Agency (EPA). Included in the RAP is this Remedial Action Selection Report (RAS), which has been developed to serve a two-fold purpose. First, it describes the activities that have been conducted by the DOE to accomplish remediation and long-term stabilization and control of the radioactive materials at the inactive uranium mill processing site near Durango, Colorado. Secondly, this document and the rest of the RAP, upon concurrence and execution by the DOE, the State of Colorado, and the NRC, become Appendix B of the Cooperative Agreement between the DOE and the State of Colorado.

  12. Remedial Action Plan and site design for stabilization of the inactive uranium mill tailings site at Durango, Colorado: Remedial action selection report. Revised final report

    SciTech Connect

    Not Available

    1991-12-01

    The uranium mill tailings site near Durango, Colorado, was one of 24 inactive uranium mill sites designated to be remediated by the US Department of Energy (DOE) under the Uranium Mill Tailings Radiation Control Act of 1978 (UMTRCA). Part of the UMTRCA requires that the US Nuclear Regulatory Commission (NRC) concur with the DOE`s Remedial Action Plan (RAP) and certify that the remedial action conducted at the site complies with the standards promulgated by the US Environmental Protection Agency (EPA). Included in the RAP is this Remedial Action Selection Report (RAS), which has been developed to serve a two-fold purpose. First, it describes the activities that have been conducted by the DOE to accomplish remediation and long-term stabilization and control of the radioactive materials at the inactive uranium mill processing site near Durango, Colorado. Secondly, this document and the rest of the RAP, upon concurrence and execution by the DOE, the State of Colorado, and the NRC, become Appendix B of the Cooperative Agreement between the DOE and the State of Colorado.

  13. Remedial Action Plan and site design for stabilization of the inactive uranium mill tailings site at Falls City, Texas. [Uranium Mill Tailings Remedial Action (UMTRA) Project

    SciTech Connect

    Chernoff, A.R. . Uranium Mill Tailings Remedial Action Project Office); Lacker, D.K. . Bureau of Radiation Control)

    1992-09-01

    The uranium processing site near Falls City, Texas, was one of 24 inactive uranium mill sites designated to be remediated by the US Department of Energy (DOE) under Title I of the Uranium Mill Tailings Radiation Control Act of 1978 (UMTRCA). The UMTRCA requires that the US Nuclear Regulatory Commission (NRC) concur with the DOE's remedial action plan (RAP) and certify that the remedial action conducted at the site complies with the standards promulgated by the US Environmental Protection Agency (EPA). The RAP, which includes this summary remedial action selection report (RAS), serves a two-fold purpose. First, it describes the activities proposed by the DOE to accomplish long-term stabilization and control of the residual radioactive materials at the inactive uranium processing site near Falls City, Texas. Second, this document and the remainder of the RAP, upon concurrence and execution by the DOE, the State of Texas, and the NRC, becomes Appendix B of the Cooperative Agreement between the DOE and the State of Texas.

  14. Remedial action plan and site design for stabilization of the inactive uranium processing site at Naturita, Colorado. Remedial action selection report, Attachment 2, Geology report: Preliminary final

    SciTech Connect

    Not Available

    1993-08-01

    The uranium processing site near Naturita, Colorado, is one of 24 inactive uranium mill sites designated to be cleaned up by the US Department of Energy (DOE) under the Uranium Mill Tailings Radiation Control Act of 1978 (UMTRCA), Public Law 95-604. Part of the UMTRCA requires that the US Nuclear Regulatory Commission (NRC) concur with the DOE`s remedial action plan (RAP) and certify that the remedial action conducted at the site complies with the standards promulgated by the US Environmental Protection Agency (EPA). Included in the RAP is this Remedial Action Selection Report (RAS), which serves two purposes. First, it describes the activities that are proposed by the DOE to accomplish remediation and long-term stabilization and control of the radioactive materials at the inactive uranium processing site near Naturita, Colorado. Second, this document and the rest of the RAP, upon concurrence and execution by the DOE, the state of Colorado, and the NRC, become Appendix B of the cooperative agreement between the DOE and the State of Colorado.

  15. A terrain-based paired-site sampling design to assess biodiversity losses from eastern hemlock decline.

    PubMed

    Young, John A; Smith, David R; Snyder, Craig D; Lemarie, David P

    2002-06-01

    Biodiversity surveys are often hampered by the inability to control extraneous sources of variability introduced into comparisons of populations across a heterogenous landscape. If not specifically accounted for a priori, this noise can weaken comparisons between sites, and can make it difficult to draw inferences about specific ecological processes. We developed a terrain-based, paired-site sampling design to analyze differences in aquatic biodiversity between streams draining eastern hemlock (Tsuga canadensis) forests, and those draining mixed hardwood forests in Delaware Water Gap National Recreation Area (USA). The goal of this design was to minimize variance due to terrain influences on stream communities, while representing the range of hemlock dominated stream environments present in the park. We used geographic information systems (GIS) and cluster analysis to define and partition hemlock dominated streams into terrain types based on topographic variables and stream order. We computed similarity of forest stands within terrain types and used this information to pair hemlock-dominated streams with hardwood counterparts prior to sampling. We evaluated the effectiveness of the design through power analysis and found that power to detect differences in aquatic invertebrate taxa richness was highest when sites were paired and terrain type was included as a factor in the analysis. Precision of the estimated difference in mean richness was nearly doubled using the terrain-based, paired site design in comparison to other evaluated designs. Use of this method allowed us to sample stream communities representative of park-wide forest conditions while effectively controlling for landscape variability.

  16. A terrain-based paired-site sampling design to assess biodiversity losses from eastern hemlock decline

    USGS Publications Warehouse

    Young, J.A.; Smith, D.R.; Snyder, C.D.; Lemarie, D.P.

    2002-01-01

    Biodiversity surveys are often hampered by the inability to control extraneous sources of variability introduced into comparisons of populations across a heterogenous landscape. If not specifically accounted for a priori, this noise can weaken comparisons between sites, and can make it difficult to draw inferences about specific ecological processes. We developed a terrain-based, paired-site sampling design to analyze differences in aquatic biodiversity between streams draining eastern hemlock (Tsuga canadensis) forests, and those draining mixed hardwood forests in Delaware Water Gap National Recreation Area (USA). The goal of this design was to minimize variance due to terrain influences on stream communities, while representing the range of hemlock dominated stream environments present in the park. We used geographic information systems (GIS) and cluster analysis to define and partition hemlock dominated streams into terrain types based on topographic variables and stream order. We computed similarity of forest stands within terrain types and used this information to pair hemlock-dominated streams with hardwood counterparts prior to sampling. We evaluated the effectiveness of the design through power analysis and found that power to detect differences in aquatic invertebrate taxa richness was highest when sites were paired and terrain type was included as a factor in the analysis. Precision of the estimated difference in mean richness was nearly doubled using the terrain-based, paired site design in comparison to other evaluated designs. Use of this method allowed us to sample stream communities representative of park-wide forest conditions while effectively controlling for landscape variability.

  17. Multistepped Drill Design for Single-Stage Implant Site Preparation: Experimental Study in Type 2 Bone.

    PubMed

    Abboud, Marcus; Delgado-Ruiz, Rafael Arcesio; Kucine, Allan; Rugova, Sihana; Balanta, Julian; Calvo-Guirado, Jose Luis

    2015-10-01

    To evaluate an experimental multistepped drill for single-stage implant site preparation by means of real-time analysis of thermal variations during and postdrilling, and by implant stability evaluation. Temperature and time were recorded in real time by paired microprobe thermocouples during simulated osteotomy in type 2 bone similes at the cortical and cancellous zones. Three different drilling groups with a new multistepped drill design were compared: Control (2-mm diameter pilot drill + 3.3-mm three-stepped drill + 4.1-mm three-stepped drill); Test A (3.3-mm three-stepped drill); and Test B (4.1-mm three-stepped drill). Implants were inserted, and implant stability was evaluated with the Perio Test Value (PTV). Two-way anova was used to test the independent effects of osteotomy and implant diameter on temperature and stability. All the drills induced thermal changes without significant differences between groups (p > .05). Drilling in cortical bone produced significant increase of the temperatures in a range of 1.8 ± 0.9°C compared with drilling in cancellous bone (p < .05). ΔT temperatures were significantly higher for test groups in cortical and cancellous bone (p < .05); ΔT10 for all groups showed a reduction of the temperature in a range of 1.7 ± 0.3°C without significant differences between groups (p > .05); the mean time to accomplish drilling was significantly longer in the control group (p < .05); test groups took 10 ± 0.3 seconds less to reach the required drilling depth. PTV values were higher in test groups compared with controls (p < .05). The multistepped drills used for single-stage implant site preparation Increase temperature as in comparison with a conventional incremental protocol; Induce the temperature increment in cortical bone compared with the cancellous bone; Reduce drilling time when a multistepped drill is used alone; and Increase implant stability twofold compared with a conventional

  18. Human Apurinic/Apyrimidinic Endonuclease siRNA Inhibits the Angiogenesis Induced by X-Ray Irradiation in Lung Cancer Cells

    PubMed Central

    Gu, Xianqing; Cun, Yanping; Li, Mengxia; Qing, Yi; Jin, Feng; Zhong, Zhaoyang; Dai, Nan; Qian, Chengyuan; Sui, Jiangdong; Wang, Dong

    2013-01-01

    Objective: Radiotherapy is an important and effective treatment method for non-small cell lung cancer (NSCLC). Nonetheless, radiotherapy can alter the expression of proangiogenic molecules and induce angiogenesis. Human apurinic/apyrimidinic endonuclease (APE1) is a multifunctional protein, which has DNA repair and redox function. Our previous studies indicated APE1 is also a crucial angiogenic regulator. Thus, we investigated the effect of APE1 on radiation-induced angiogenesis in lung cancer and its underlying mechanism. Methods: Tumor specimens of 136 patients with NSCLC were obtained from 2003 to 2008. The APE1 and vascular endothelial growth factor (VEGF) expression, as well as microvessel density (MVD) were observed with immunohistochemistry in tumor samples. Human lung adenocarcinoma A549 cells were treated with Ad5/F35-APE1 siRNA and/or irradiation, and then the cells were used for APE1 analysis by Western blot and VEGF analysis by RT-PCR and ELISA. To elucidate the underline mechanism of APE1 on VEGF expression, HIF-1α protein level was determined by Western blot, and the DNA binding activity of HIF-1α was detected by EMSA. Transwell migration assay and capillary-like structure assay were used to observe the migration and capillary-like structure formation ability of human umbilical veins endothelial cells (HUVECs) that were co-cultured with Ad5/F35-APE1 siRNA and (or) irradiation treated A549 cells culture medium. Results: The high expression rates of APE1 and VEGF in NSCLC were 77.94% and 66.18%, respectively. The expressions of APE1 was significantly correlated with VEGF and MVD (r=0.369, r=0.387). APE1 and VEGF high expression were significantly associated with reduced disease free survival (DFS) time. The high expressions of APE1 and VEGF on A549 cells were concurrently induced by X-ray irradiation in a dose-dependent manner. Silencing of APE1 by Ad5/F35-APE1 siRNA significantly decreased DNA binding activity of HIF-1α and suppressed the expression

  19. LEAD (Pb) IN BIOTA AND PERCEPTIONS OF Pb EXPOSURE AT A RECENTLY DESIGNATED SUPERFUND BEACH SITE IN NEW JERSEY

    PubMed Central

    Burger, Joanna; Gochfeld, Michael; Jeitner, Christian; Donio, Mark; Pittfield, Taryn

    2014-01-01

    The Raritan Bay Slag Site (New Jersey) was designated a Superfund site in 2009 because the seawall, jetties, and sediment contained lead (Pb). Our objective was to compare Pb and mercury (Hg) levels in biota and public perceptions of exposure at the Superfund and reference sites. Samples (algae, invertebrates, fish) were collected from the Raritan Bay Slag Site and reference sites and analyzed for Pb and Hg. Waterfront users were interviewed using a standard questionnaire. Levels of Pb in aquatic organisms were compared to ecological and human health safety standards. Lead levels were related to location, trophic level, and mobility. Lead levels in biota were highest at the western side of the West Jetty. Mean Pb levels were highest for algae (Fucus = 53,600 ± 6990 ng/g = ppb [wet weight], Ulva = 23,900 ± 2430 ppb), intermediate for grass shrimp (7270 ± 1300 ppb, 11,600 ± 3340 ppb), and lowest for fish (Atlantic silversides 218 ± 44 ppb). Within species, Pb levels varied significantly across the sampling sites. Lead levels in algae, sometimes ingested by individuals, were sufficiently high to exceed human safety levels. Mercury levels did not differ between the Superfund and reference sites. Despite the fence and warnings, people (1) used the Superfund and reference sites similarly, (2) had similar fish consumption rates, and (3) were not concerned about Pb, although most individuals knew the metal was present. The fish sampled posed no apparent risk for human consumers, but the algae did. PMID:22409490

  20. Lead (Pb) in biota and perceptions of Pb exposure at a recently designated Superfund beach site in New Jersey.

    PubMed

    Burger, Joanna; Gochfeld, Michael; Jeitner, Christian; Donio, Mark; Pittfield, Taryn

    2012-01-01

    The Raritan Bay Slag Site (New Jersey) was designated a Superfund site in 2009 because the seawall, jetties, and sediment contained lead (Pb). Our objective was to compare Pb and mercury (Hg) levels in biota and public perceptions of exposure at the Superfund and reference sites. Samples (algae, invertebrates, fish) were collected from the Raritan Bay Slag Site and reference sites and analyzed for Pb and Hg. Waterfront users were interviewed using a standard questionnaire. Levels of Pb in aquatic organisms were compared to ecological and human health safety standards. Lead levels were related to location, trophic level, and mobility. Lead levels in biota were highest at the western side of the West Jetty. Mean Pb levels were highest for algae (Fucus = 53,600 ± 6990 ng/g = ppb [wet weight], Ulva = 23,900 ± 2430 ppb), intermediate for grass shrimp (7270 ± 1300 ppb, 11,600 ± 3340 ppb), and lowest for fish (Atlantic silversides 218 ± 44 ppb). Within species, Pb levels varied significantly across the sampling sites. Lead levels in algae, sometimes ingested by individuals, were sufficiently high to exceed human safety levels. Mercury levels did not differ between the Superfund and reference sites. Despite the fence and warnings, people (1) used the Superfund and reference sites similarly, (2) had similar fish consumption rates, and (3) were not concerned about Pb, although most individuals knew the metal was present. The fish sampled posed no apparent risk for human consumers, but the algae did.

  1. Preliminary thermal/thermomechanical analyses of the Site Characterization Plan's Conceptual Design for a repository containing horizontally emplaced waste packages at the Deaf Smith County site

    SciTech Connect

    Ghantous, N.Y.; Raines, G.E.

    1987-10-01

    This report presents thermal/thermomechanical analyses of the Site Characterization Plan Conceptual Design for horizontal package emplacement at the Deaf Smith County site, Texas. The repository was divided into three geometric regions. Then two-dimensional finite-element models were set up to approximate the three-dimensional nature of each region. Thermal and quasistatic thermomechanical finite-element analyses were performed to evaluate the thermal/thermomechanical responses of the three regions. The exponential-time creep law was used to represent the creep behavior of salt rock. The repository design was evaluated by comparing the thermal/thermomechanical responses obtained for the three regions with interim performance constraints. The preliminary results show that all the performance constraints are met except for those of the waste package. The following factors were considered in interpreting these results: (1) the qualitative description of the analytical responses; (2) the limitations of the analyses; and (3) either the conclusions based on overall evaluation of limitations and analytical results or the conclusions based on the fact that the repository design may be evaluated only after further analyses. Furthermore, a parametric analysis was performed to estimate the effect of material parameters on the predicted thermal/thermomechanical response. 23 refs., 34 figs., 9 tabs.

  2. Hydration in drug design. 3. Conserved water molecules at the ligand-binding sites of homologous proteins.

    PubMed

    Poornima, C S; Dean, P M

    1995-12-01

    Water molecules are known to play an important rôle in mediating protein-ligand interactions. If water molecules are conserved at the ligand-binding sites of homologous proteins, such a finding may suggest the structural importance of water molecules in ligand binding. Structurally conserved water molecules change the conventional definition of 'binding sites' by changing the shape and complementarity of these sites. Such conserved water molecules can be important for site-directed ligand/drug design. Therefore, five different sets of homologous protein/protein-ligand complexes have been examined to identify the conserved water molecules at the ligand-binding sites. Our analysis reveals that there are as many as 16 conserved water molecules at the FAD binding site of glutathione reductase between the crystal structures obtained from human and E. coli. In the remaining four sets of high-resolution crystal structures, 2-4 water molecules have been found to be conserved at the ligand-binding sites. The majority of these conserved water molecules are either bound in deep grooves at the protein-ligand interface or completely buried in cavities between the protein and the ligand. All these water molecules, conserved between the protein/protein-ligand complexes from different species, have identical or similar apolar and polar interactions in a given set. The site residues interacting with the conserved water molecules at the ligand-binding sites have been found to be highly conserved among proteins from different species; they are more conserved compared to the other site residues interacting with the ligand. These water molecules, in general, make multiple polar contacts with protein-site residues.

  3. Assessment, approval, design and construction of a facility on a Superfund site in 36 months

    SciTech Connect

    Drag, D.J.; Webb, C.K.; Luenenborg, G.W.

    1996-11-01

    The Union Pacific Resources Corporation (UPRC) owned and operated a 600-acre crude oil production field in Wilmington, California. UPRC granted a lease to the TCL Corporation in 1951 for the disposal of oil and gas drilling field wastes. In the 1950s and 1960s, waste materials consisting of oil-free rotary mud, as well as rotary mud containing oil and crude oil tank bottoms were accepted at the site. Site testing and record investigations have shown that some other wastes, inconsistent with those permitted by the agreement between UPRC and TCL, were also disposed of at the site. Soil samples collected from the site in 1981 showed moderately high levels of metals in the soils at the site. In 1983, the site was included on the California State Superfund list of hazardous waste contaminated sites. In 1988, UPRC signed a Consent order Agreement with the Department of Toxic Substance Control (DTSC) whereby UPRC agreed to investigate a 31-acre area of the Study Area and develop a Remedial Action Plan (RAP) with DTSC oversight. The Port of Long Beach (POLB) is assuming the responsibility for implementing subsequent investigative and remedial activities at the portions of the site which they have since purchased from UPRC. The overall project objective was to investigate a 31-acre parcel within the Study Area (TCL site), develop a RAP, remediate the oil sump soils, and develop the parcel as an automobile distribution facility to be used by Toyota Motor Sales (TMS).

  4. Designing personable and informative job recruiting web sites: testing the effect of the design on attractiveness and intent to apply.

    PubMed

    Thoms, Peg; Chinn, Susan J; Goodrich, Jana; Howard, Gary

    2004-06-01

    To date, most of the limited research conducted about the efficacy of corporate job recruiting web sites has been either anecdotal or based on field surveys. In this study, the effects of using a photograph, a friendly text message, and a list of job tasks in job descriptions were measured on undergraduates' ratings of the Personableness and Informativeness of recruiting web sites. In addition, the relationships between ratings of Personableness and Informativeness on perceptions of organizational Attractiveness and Intent to Apply were tested. Use of a photograph increased ratings of both Personableness and Informativeness. Additional results are provided and directions for research suggested.

  5. Benchmarking a computational design method for the incorporation of metal ion-binding sites at symmetric protein interfaces.

    PubMed

    Hansen, William A; Khare, Sagar D

    2017-08-01

    The design of novel metal-ion binding sites along symmetric axes in protein oligomers could provide new avenues for metalloenzyme design, construction of protein-based nanomaterials and novel ion transport systems. Here, we describe a computational design method, symmetric protein recursive ion-cofactor sampling (SyPRIS), for locating constellations of backbone positions within oligomeric protein structures that are capable of supporting desired symmetrically coordinated metal ion(s) chelated by sidechains (chelant model). Using SyPRIS on a curated benchmark set of protein structures with symmetric metal binding sites, we found high recovery of native metal coordinating rotamers: in 65 of the 67 (97.0%) cases, native rotamers featured in the best scoring model while in the remaining cases native rotamers were found within the top three scoring models. In a second test, chelant models were crossmatched against protein structures with identical cyclic symmetry. In addition to recovering all native placements, 10.4% (8939/86013) of the non-native placements, had acceptable geometric compatibility scores. Discrimination between native and non-native metal site placements was further enhanced upon constrained energy minimization using the Rosetta energy function. Upon sequence design of the surrounding first-shell residues, we found further stabilization of native placements and a small but significant (1.7%) number of non-native placement-based sites with favorable Rosetta energies, indicating their designability in existing protein interfaces. The generality of the SyPRIS approach allows design of novel symmetric metal sites including with non-natural amino acid sidechains, and should enable the predictive incorporation of a variety of metal-containing cofactors at symmetric protein interfaces. © 2017 The Protein Society.

  6. Mapping polyamide-DNA interactions in human cells reveals a new design strategy for effective targeting of genomic sites.

    PubMed

    Erwin, Graham S; Bhimsaria, Devesh; Eguchi, Asuka; Ansari, Aseem Z

    2014-09-15

    Targeting the genome with sequence-specific synthetic molecules is a major goal at the interface of chemistry, biology, and personalized medicine. Pyrrole/imidazole-based polyamides can be rationally designed to target specific DNA sequences with exquisite precision in vitro; yet, the biological outcomes are often difficult to interpret using current models of binding energetics. To directly identify the binding sites of polyamides across the genome, we designed, synthesized, and tested polyamide derivatives that enabled covalent crosslinking and localization of polyamide-DNA interaction sites in live human cells. Bioinformatic analysis of the data reveals that clustered binding sites, spanning a broad range of affinities, best predict occupancy in cells. In contrast to the prevailing paradigm of targeting single high-affinity sites, our results point to a new design principle to deploy polyamides and perhaps other synthetic molecules to effectively target desired genomic sites in vivo. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Development of in-structure design spectra for dome mounted equipment on underground waste storage tanks at the Hanford Site

    SciTech Connect

    Julyk, L.J.

    1995-09-01

    In-structure response spectra for dome mounted equipment on underground waste storage tanks at the Hanford Site are developed on the basis of recent soil-structure-interaction analyses. Recommended design spectra are provided for various locations on the tank dome.

  8. User-Interface Design Characteristics of Fortune 500 B2C E-Commerce Sites and Industry Differences

    ERIC Educational Resources Information Center

    Zhao, Jensen J.; Truell, Allen D.; Alexander, Melody W.

    2006-01-01

    This study examined the user-interface design characteristics of 107 Fortune 500 B2C e-commerce Web sites and industry differences. Data were collected from corporate homepages, B2C product/service pages, B2C interactive shopping pages, as well as customer satisfaction of 321 online shoppers. The findings indicate that (a) to attract online…

  9. 41 CFR 102-76.55 - What sustainable development principles must Federal agencies apply to the siting, design, and...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... and Construction Sustainable Development § 102-76.55 What sustainable development principles must... Acquisition,” Federal agencies must apply sustainable development principles to the siting, design, and... 41 Public Contracts and Property Management 3 2010-07-01 2010-07-01 false What...

  10. Long-Range School Site Location Plan: PROJECT DESIGN. Interagency Planning for Urban Educational Needs, Number 36.

    ERIC Educational Resources Information Center

    Tokmakian, Harold

    This report, written as part of Phase II of PROJECT DESIGN, an ESEA Title III project administered by the Fresno City Unified School District, contains an analysis of the factors that must be considered in locating new school sites for the Fresno district as well as the principles and standards related to community planning and school site…

  11. Independent Design Review: Grants Chlorinated Solvents Plume, Superfund Site, Grants, Cibola County, New Mexico, EPA Region 6

    EPA Pesticide Factsheets

    The Grants Chlorinated Solvents Plume Superfund Site in Grants, Cibola County, New Mexico was selected by EPA OSRTI based on a nomination from EPA Region 6. The remedy is in the early design stage and has an estimated cost of $29.5 million.

  12. User-Interface Design Characteristics of Fortune 500 B2C E-Commerce Sites and Industry Differences

    ERIC Educational Resources Information Center

    Zhao, Jensen J.; Truell, Allen D.; Alexander, Melody W.

    2006-01-01

    This study examined the user-interface design characteristics of 107 Fortune 500 B2C e-commerce Web sites and industry differences. Data were collected from corporate homepages, B2C product/service pages, B2C interactive shopping pages, as well as customer satisfaction of 321 online shoppers. The findings indicate that (a) to attract online…

  13. 41 CFR 102-76.20 - What issues must Federal agencies consider in providing site planning and landscape design services?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 41 Public Contracts and Property Management 3 2013-07-01 2013-07-01 false What issues must Federal... What issues must Federal agencies consider in providing site planning and landscape design services? In... architectural characteristics of the building; (f) Comply with the requirements of the National Environmental...

  14. Making the Most of Your School Site. School Buildings Planning, Design, and Construction Series No. 2.

    ERIC Educational Resources Information Center

    Odell, John H.

    A school construction guide offers key personnel in school development projects information on the complex task of master planning and construction of schools in Australia. This chapter of the guide provides advice on site selection covering selection criteria; traffic issues; and site services, such as water, power, and sewer. Additionally…

  15. Remedial action plan and site design for stabilization of the inactive Uranium Mill Tailings Site at Lowman, Idaho. Attachment 4, Water resources protection strategy: Final report

    SciTech Connect

    Not Available

    1991-09-01

    The DOE proposes to achieve compliance with the proposed EPA groundwater protection standards (Subparts A and B of 40 CFR 192) by meeting the EPA maximum concentration limits (MCLs) or background concentrations for designated hazardous constituents in groundwater in the uppermost aquifer (alluvium/weathered granodiorite) at the point of compliance (POC) at the Lowman disposal site near Lowman, Idaho. The proposed remedial action in conjunction with existing hydrogeological conditions at the Lowman site will ensure sufficient protection of human health and the environment. The DOE has concluded that the EPA groundwater protection standards will be met at the POC because, with the exception of antimony, none of the hazardous constituents that exceed laboratory method detection limits within the radioactive sand pore fluids were above the proposed concentration limits. The DOE has demonstrated that antimony will meet the proposed concentration limits at the POC through attenuation in subsoils beneath the disposal cell and by dilution in groundwater underflow. The Lowman processing site is in compliance with Subpart B of 40 CFR 192 because statistical analyses of groundwater samples indicate no groundwater contamination.

  16. Examination of CRISPR/Cas9 design tools and the effect of target site accessibility on Cas9 activity.

    PubMed

    Lee, Ciaran M; Davis, Timothy H; Bao, Gang

    2017-03-16

    The recent adaptation of the CRISPR/Cas9 system for targeted genome engineering has led to its widespread applications in many fields worldwide. In order to better understand the design rules of CRISPR/Cas9 systems, several groups have carried out large library-based screens leading to some insight into sequence preferences among highly active target sites. To facilitate CRISPR/Cas9 design these studies have spawned a plethora of gRNA design tools with algorithms based solely on direct or indirect sequence features. Here we demonstrate that the predictive power of these tools is poor, suggesting that sequence features alone cannot accurately inform the cutting efficiency of a particular CRISPR/Cas9 gRNA design. Furthermore we demonstrate that DNA target site accessibility influences the activity of CRISPR/Cas9. With further optimisation we hypothesise that it will be possible to increase the predictive power of gRNA design tools by including both sequence and target site accessibility metrics. This article is protected by copyright. All rights reserved.

  17. Remedial Action Plan and site design for stabilization of the inactive uranium mill tailings site at Mexican Hat, Utah: Text, Appendices A--C. Final report

    SciTech Connect

    1988-07-01

    This Remedial Action Plan (RAP) has been developed to serve a two- fold purpose. It presents the activities proposed by the Department of Energy (DOE) to accomplish long-term stabilization and control of the residual radioactive materials (RRM) from Monument Valley Arizona, and Mexican Hat, Utah, at the Mexican Hat disposal site. It also serves to document the concurrence of both the Navajo Nation and the Nuclear Regulatory Commission (NRC) in the remedial action. This agreement, upon execution by DOE and the Navajo Nation and concurrence by the NRC, becomes Appendix B of the Cooperative Agreement. This document has been structured to provide a comprehensive understanding of the remedial action proposed for the Monument Valley and Mexican Hat sites. It includes specific design and construction requirements for the remedial action. Pertinent information and data are included with reference given to the supporting documents. Appendices A,B, and C are provided as part of this document. Appendix A presents regulatory compliance issues, Appendix B provides details of the engineering design, and Appendix C presents the radiological support plan.

  18. Data for the geochemical investigation of UMTRAP designated site at Ambrosia Lake, New Mexico

    SciTech Connect

    Markos, G.; Bush, K.J.

    1983-09-01

    This report contains the geochemical data and the methods of data collection from the former tailings site at Ambrosia Lake, New Mexico. Data are from a one-time sampling of waters and solid material from the background, the area adjacent to the site, and the site. Selected solid samples are water extracted to remove easily soluble salts and acid extracted to remove carbonates and hydroxides. The waters, extracts, and solid samples were analyzed for selected major and trace elements. 3 refs., 2 figs., 1 tab.

  19. Data for the geochemical investigation of UMTRAP designated site at Tuba City, Arizona

    SciTech Connect

    Markos, G.; Bush, K.J.

    1983-09-01

    This report contains the geochemical data and the methods of data collection from the former tailings site at Tuba City, Arizona. Data are from a one-time sampling of waters and solid material from the background, the area adjacent to the site, and the site. Selected solid samples are water extracted to remove easily soluble salts and acid extracted to remove carbonates and hydroxides. The waters, extracts, and solid samples were analyzed for selected major and trace elements. 4 refs., 2 figs., 1 tab.

  20. Rational design of a functional metalloenzyme: introduction of a site for manganese binding and oxidation into a heme peroxidase.

    PubMed

    Wilcox, S K; Putnam, C D; Sastry, M; Blankenship, J; Chazin, W J; McRee, D E; Goodin, D B

    1998-12-01

    The design of a series of functionally active models for manganese peroxidase (MnP) is described. Artificial metal binding sites were created near the heme of cytochrome c peroxidase (CCP) such that one of the heme propionates could serve as a metal ligand. At least two of these designs, MP6.1 and MP6.8, bind Mn2+ with Kd congruent with 0.2 mM, react with H2O2 to form stable ferryl heme species, and catalyze the steady-state oxidation of Mn2+ at enhanced rates relative to WT CCP. The kinetic parameters for this activity vary considerably in the presence of various dicarboxylic acid chelators, suggesting that the similar features displayed by native MnP are largely intrinsic to the manganese oxidation reaction rather than due to a specific interaction between the chelator and enzyme. Analysis of pre-steady-state data shows that electron transfer from Mn2+ to both the Trp-191 radical and the ferryl heme center of compound ES is enhanced by the metal site mutations, with transfer to the ferryl center showing the greatest stimulation. These properties are perplexingly similar to those reported for an alternate model for this site (1), despite rather distinct features of the two designs. Finally, we have determined the crystal structure at 1.9 A of one of our designs, MP6.8, in the presence of MnSO4. A weakly occupied metal at the designed site appears to coordinate two of the proposed ligands, Asp-45 and the heme 7-propionate. Paramagnetic nuclear magnetic resonance spectra also suggest that Mn2+ is interacting with the heme 7-propionate in MP6.8. The structure provides a basis for understanding the similar results of Yeung et al. (1), and suggests improvements for future designs.

  1. Site characterization plan: Conceptual design report: Volume 4, Appendices F-O: Nevada Nuclear Waste Storage Investigations Project

    SciTech Connect

    MacDougall, H R; Scully, L W; Tillerson, J R

    1987-09-01

    The site for the prospective repository is located at Yucca Mountain in southwestern Nevada, and the waste emplacement area will be constructed in the underlying volcanic tuffs. The target horizon for waste emplacement is a sloping bed of densely welded tuff more than 650 ft below the surface and typically more than 600 ft above the water table. The conceptual design described in this report is unique among repository designs in that it uses ramps in addition to shafts to gain access to the underground facility, the emplacement horizon is located above the water table, and it is possible that 300- to 400-ft-long horizontal waste emplacement boreholes will be used. This report summarizes the design bases, design and performance criteria, and the design analyses performed. The current status of meeting the preclosure performance objectives for licensing and of resolving the repository design and preclosure issues is presented. The repository design presented in this report will be expanded and refined during the advanced conceptual design, the license application design, and the final procurement and construction design phases. Volume 4 contains Appendices F to O.

  2. 40 CFR 228.15 - Dumping sites designated on a final basis.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Rockaway Inlet, Long Island, New York. (5) Shark River, New Jersey Dredged Material Disposal Site. (i...: Disposal shall be limited to dredged material from Shark River Inlet, New Jersey. (6) Historical Area...

  3. System-wide design issues for the stellar interferometer technology experiment (SITE)

    NASA Astrophysics Data System (ADS)

    Miller, David W.; Crawford, Samuel L.; Hyde, Tristram T.; Masters, B. P.; Crawley, Edward F.; Blackwood, Gary H.; Colavita, M. Mark; Yu, Jeffrey W.; Shao, Michael; Laskin, Robert A.

    1995-06-01

    The Stellar Interferometer Technology Experiment (SITE) is a near-term precursor mission for spaceborne optical interferometry. Proposed by the MIT Space Engineering Research Center and NASA's Jet Propulsion Laboratory, SITE is a two-aperture stellar interferometer located in the payload bay of the Space Shuttle. It has a baseline of four meters, operates with a detection bandwidth of 300 nanometers in the visible spectrum, and consists of three optical benches kinematically mounted inside a precision truss structure. The objective of SITE is to demonstrate system-level functionality of a space-based stellar interferometer through the use of enabling and enhancing Controlled Structures Technologies such as vibration isolation and suppression. Moreover, SITE will validate, in the space environment, technologies such as optical delay lines, laser metrology systems, fringe detectors, active fringe trackers, and high- bandwidth pointing control systems which are critical for realizing future space-based astrometric and imaging interferometers.

  4. Final Report: Independent Design Review, Celanese Fiber Operations Superfund Site, Shelby, North Carolina, EPA Region 4

    EPA Pesticide Factsheets

    The Celanese site is located in Shelby, North Carolina. Operation of the P&T system was discontinued on a trial basis for two years to evaluate monitored natural attenuation as a potential ground water remedy.

  5. 24 CFR 3285.103 - Site suitability with design zone maps.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... is to be installed. The design zone maps are those identified in part 3280 of this chapter. (a) Wind zone. Manufactured homes must not be installed in a wind zone that exceeds the design wind loads for which the home has been designed, as evidenced by the wind zone indicated on the home's data plate and...

  6. Cognitive Issues in the Design and Deployment of Interactive Hypermedia: Implications for Authoring WWW Sites.

    ERIC Educational Resources Information Center

    Ebersole, Samuel

    1997-01-01

    In designing effective interactive media, cognitive psychology, human factors, psycholinguistics, semiotics, and communication theory should be considered. This article defines interactive media, considers interactive hypermedia design and the cognitive processes of designer and user, and examines the World Wide Web and Netscape Navigator, a…

  7. Trends in Culturally Relevant Interface Design Features for Latino Web Site Users

    ERIC Educational Resources Information Center

    Sachau, Lori L.; Hutchinson, Susan R.

    2012-01-01

    There is a lack of published research on designing Web-based instruction for the adult U.S. Latino population. Instructional designers need guidance on how to design culturally relevant learning environments for this audience, particularly for Latino people from Mexican heritage. The authors used content analysis to investigate the extent to which…

  8. Trends in Culturally Relevant Interface Design Features for Latino Web Site Users

    ERIC Educational Resources Information Center

    Sachau, Lori L.; Hutchinson, Susan R.

    2012-01-01

    There is a lack of published research on designing Web-based instruction for the adult U.S. Latino population. Instructional designers need guidance on how to design culturally relevant learning environments for this audience, particularly for Latino people from Mexican heritage. The authors used content analysis to investigate the extent to which…

  9. Site-specific investigations and preliminary design for the Gorleben repository

    SciTech Connect

    Brennecke, P.; Kranz, H.; Schneider, H. )

    1992-01-01

    In the Federal Republic of Germany, the disposal of radioactive waste is being planned for deep geologic formations only. A repository will be constructed in the Gorleben salt dome for all kinds of radioactive waste, in particular, heat-generating waste from reprocessing and spent fuel. Pusuant to German safety criteria, a detailed site characterization program including above-ground and underground investigations must be performed to provide all necessary data for the site-specific safety assessment.

  10. Self-catalyzed site-specific depurination of G residues mediated by cruciform extrusion in closed circular DNA plasmids.

    PubMed

    Amosova, Olga; Kumar, Veena; Deutsch, Aaron; Fresco, Jacques R

    2011-10-21

    A major variety of "spontaneous" genomic damage is endogenous generation of apurinic sites. Depurination rates vary widely across genomes, occurring with higher frequency at "depurination hot spots." Recently, we discovered a site-specific self-catalyzed depurinating activity in short (14-18 nucleotides) DNA stem-loop-forming sequences with a 5'-G(T/A)GG-3' loop and T·A or G·C as the first base pair at the base of the loop; the 5'-G residue of the loop self-depurinates at least 10(5)-fold faster than random "spontaneous" depurination at pH 5. Formation of the catalytic intermediate for self-depurination in double-stranded DNA requires a stem-loop to extrude as part of a cruciform. In this study, evidence is presented for self-catalyzed depurination mediated by cruciform formation in plasmid DNA in vitro. Cruciform extrusion was confirmed, and its extent was quantitated by digestion of the plasmid with single strand-specific mung bean endonuclease, followed by restriction digestion and sequencing of resulting mung bean-generated fragments. Appearance of the apurinic site in the self-depurinating stem-loop was confirmed by digestion of plasmid DNA with apurinic endonuclease IV, followed by primer extension and/or PCR amplification to detect the endonuclease-generated strand break and identify its location. Self-catalyzed depurination was contingent on the plasmid being supercoiled and was not observed in linearized plasmids, consistent with the presence of the extruded cruciform in the supercoiled plasmid and not in the linear one. These results indicate that self-catalyzed depurination is not unique to single-stranded DNA; rather, it can occur in stem-loop structures extruding from double-stranded DNA and therefore could, in principle, occur in vivo.

  11. Comparison of protein active site structures for functional annotation of proteins and drug design.

    PubMed

    Powers, Robert; Copeland, Jennifer C; Germer, Katherine; Mercier, Kelly A; Ramanathan, Viswanathan; Revesz, Peter

    2006-10-01

    Rapid and accurate functional assignment of novel proteins is increasing in importance, given the completion of numerous genome sequencing projects and the vastly expanding list of unannotated proteins. Traditionally, global primary-sequence and structure comparisons have been used to determine putative function. These approaches, however, do not emphasize similarities in active site configurations that are fundamental to a protein's activity and highly conserved relative to the global and more variable structural features. The Comparison of Protein Active Site Structures (CPASS) database and software enable the comparison of experimentally identified ligand-binding sites to infer biological function and aid in drug discovery. The CPASS database comprises the ligand-defined active sites identified in the protein data bank, where the CPASS program compares these ligand-defined active sites to determine sequence and structural similarity without maintaining sequence connectivity. CPASS will compare any set of ligand-defined protein active sites, irrespective of the identity of the bound ligand. Proteins 2006. (c) 2006 Wiley-Liss, Inc.

  12. ICRPfinder: a fast pattern design algorithm for coding sequences and its application in finding potential restriction enzyme recognition sites

    PubMed Central

    Li, Chao; Li, Yuhua; Zhang, Xiangmin; Stafford, Phillip; Dinu, Valentin

    2009-01-01

    Background Restriction enzymes can produce easily definable segments from DNA sequences by using a variety of cut patterns. There are, however, no software tools that can aid in gene building -- that is, modifying wild-type DNA sequences to express the same wild-type amino acid sequences but with enhanced codons, specific cut sites, unique post-translational modifications, and other engineered-in components for recombinant applications. A fast DNA pattern design algorithm, ICRPfinder, is provided in this paper and applied to find or create potential recognition sites in target coding sequences. Results ICRPfinder is applied to find or create restriction enzyme recognition sites by introducing silent mutations. The algorithm is shown capable of mapping existing cut-sites but importantly it also can generate specified new unique cut-sites within a specified region that are guaranteed not to be present elsewhere in the DNA sequence. Conclusion ICRPfinder is a powerful tool for finding or creating specific DNA patterns in a given target coding sequence. ICRPfinder finds or creates patterns, which can include restriction enzyme recognition sites, without changing the translated protein sequence. ICRPfinder is a browser-based JavaScript application and it can run on any platform, in on-line or off-line mode. PMID:19747395

  13. Catalytic residues in hydrolases: analysis of methods designed for ligand-binding site prediction

    PubMed Central

    Jadczyk, Tomasz; Roterman, Irena

    2010-01-01

    The comparison of eight tools applicable to ligand-binding site prediction is presented. The methods examined cover three types of approaches: the geometrical (CASTp, PASS, Pocket-Finder), the physicochemical (Q-SiteFinder, FOD) and the knowledge-based (ConSurf, SuMo, WebFEATURE). The accuracy of predictions was measured in reference to the catalytic residues documented in the Catalytic Site Atlas. The test was performed on a set comprising selected chains of hydrolases. The results were analysed with regard to size, polarity, secondary structure, accessible solvent area of predicted sites as well as parameters commonly used in machine learning (F-measure, MCC). The relative accuracies of predictions are presented in the ROC space, allowing determination of the optimal methods by means of the ROC convex hull. Additionally the minimum expected cost analysis was performed. Both advantages and disadvantages of the eight methods are presented. Characterization of protein chains in respect to the level of difficulty in the active site prediction is introduced. The main reasons for failures are discussed. Overall, the best performance offers SuMo followed by FOD, while Pocket-Finder is the best method among the geometrical approaches. Electronic supplementary material The online version of this article (doi:10.1007/s10822-010-9402-0) contains supplementary material, which is available to authorized users. PMID:21104192

  14. Catalytic residues in hydrolases: analysis of methods designed for ligand-binding site prediction

    NASA Astrophysics Data System (ADS)

    Prymula, Katarzyna; Jadczyk, Tomasz; Roterman, Irena

    2011-02-01

    The comparison of eight tools applicable to ligand-binding site prediction is presented. The methods examined cover three types of approaches: the geometrical (CASTp, PASS, Pocket-Finder), the physicochemical (Q-SiteFinder, FOD) and the knowledge-based (ConSurf, SuMo, WebFEATURE). The accuracy of predictions was measured in reference to the catalytic residues documented in the Catalytic Site Atlas. The test was performed on a set comprising selected chains of hydrolases. The results were analysed with regard to size, polarity, secondary structure, accessible solvent area of predicted sites as well as parameters commonly used in machine learning (F-measure, MCC). The relative accuracies of predictions are presented in the ROC space, allowing determination of the optimal methods by means of the ROC convex hull. Additionally the minimum expected cost analysis was performed. Both advantages and disadvantages of the eight methods are presented. Characterization of protein chains in respect to the level of difficulty in the active site prediction is introduced. The main reasons for failures are discussed. Overall, the best performance offers SuMo followed by FOD, while Pocket-Finder is the best method among the geometrical approaches.

  15. Work-site physical fitness programs. Comparing the impact of different program designs on cardiovascular risks.

    PubMed

    Heirich, M A; Foote, A; Erfurt, J C; Konopka, B

    1993-05-01

    The relative impact of three different approaches to physical fitness at the work site on cardiovascular risk reduction is examined, based on before/after health screening of employees, and employees' reports of participation in physical exercise activities. The three approaches tested were: 1) a staffed physical fitness facility, 2) one-to-one counseling with at-risk employees, and 3) a combination of one-to-one counseling with employees plus organization of the work site to encourage peer support and mutual exercise activity at work. A fourth site is used as a control site. The program that was centered around a physical fitness facility had little measurable impact on cardiovascular risks, and showed results similar to those at the control site. Both of the other programs were more effective, with the combination of counseling and plant organization providing the best health outcomes in terms of frequency of exercise, adequacy of blood pressure control (among hypertensives), weight loss (among the overweight), and smoking cessation. These results indicate that systematic, ongoing outreach to enlist employees in various types of exercise programs is more effective than the presence of fitness facilities without such outreach. Moreover, significant increases in frequency of exercise can be sustained without a substantial investment in facilities.

  16. Design of the aerosol sampling manifold for the Southern Great Plains site

    SciTech Connect

    Leifer, R.; Knuth, R.H.; Guggenheim, S.F.; Lee, H.N.

    1995-04-01

    To meet the needs of the ARM program, the Environmental Measurements Laboratory (EML) has the responsibility to establish a surface aerosol measurements program at the Southern Great Plains (SGP) site in Lamont, OK. At the present time, EML has scheduled installation of five instruments at SGP: a single wavelength nephelometer, an optical particle counter (OPC), a condensation particle counter (CPC), an optical absorption monitor (OAM), and an ozone monitor. ARM`s operating protocol requires that all the observational data be placed online and sent to the main computer facility in real time. EML currently maintains a computer file containing back trajectory (BT) analyses for the SGP site. These trajectories are used to characterize air mass types as they pass over the site. EML is continuing to calculate and store the resulting trajectory analyses for future use by the ARM science team.

  17. Bifunctional chelating agent for the design and development of site specific radiopharmaceuticals and biomolecule conjugation strategy

    DOEpatents

    Katti, Kattesh V.; Prabhu, Kandikere R.; Gali, Hariprasad; Pillarsetty, Nagavara Kishore; Volkert, Wynn A.

    2003-10-21

    There is provided a method of labeling a biomolecule with a transition metal or radiometal in a site specific manner to produce a diagnostic or therapeutic pharmaceutical compound by synthesizing a P.sub.2 N.sub.2 -bifunctional chelating agent intermediate, complexing the intermediate with a radio metal or a transition metal, and covalently linking the resulting metal-complexed bifunctional chelating agent with a biomolecule in a site specific manner. Also provided is a method of synthesizing the --PR.sub.2 containing biomolecules by synthesizing a P.sub.2 N.sub.2 -bifunctional chelating agent intermediate, complexing the intermediate with a radiometal or a transition metal, and covalently linking the resulting radio metal-complexed bifunctional chelating agent with a biomolecule in a site specific manner. There is provided a therapeutic or diagnostic agent comprising a --PR.sub.2 containing biomolecule.

  18. The design and implementation of web mining in web sites security

    NASA Astrophysics Data System (ADS)

    Li, Jian; Zhang, Guo-Yin; Gu, Guo-Chang; Li, Jian-Li

    2003-06-01

    The backdoor or information leak of Web servers can be detected by using Web Mining techniques on some abnormal Web log and Web application log data. The security of Web servers can be enhanced and the damage of illegal access can be avoided. Firstly, the system for discovering the patterns of information leakages in CGI scripts from Web log data was proposed. Secondly, those patterns for system administrators to modify their codes and enhance their Web site security were provided. The following aspects were described: one is to combine web application log with web log to extract more information, so web data mining could be used to mine web log for discovering the information that firewall and Information Detection System cannot find. Another approach is to propose an operation module of web site to enhance Web site security. In cluster server session, Density-Based Clustering technique is used to reduce resource cost and obtain better efficiency.

  19. A new design of highly reversible LiNiO2: Defect formation in transition metal site

    NASA Astrophysics Data System (ADS)

    Kobayashi, Yo; Tabuchi, Mitsuharu; Miyashiro, Hajime; Kuriyama, Nobuhiro

    2017-10-01

    LiNiO2 with Ni defects in the 3b sites (named "Ni defected" LiNiO2) was prepared and characterized using electrochemical calorimetry. Different from conventionally prepared "Standard" LiNiO2, "Ni defected" LiNiO2 exhibited high reversibility. The 1st coulombic efficiency was recorded to be 101% due to minimization from the Ni ion substitution at the Li 3a sites. The reversible capacity was 212 mAh g-1 in the 1st cycle with 87% capacity retention after 80 cycles because Li+ ordering and Jahn-Teller distortion during the charge and discharge cycle were suppressed by introducing the Ni defect 3b sites. The proposed simple defect design concept introduced the development of a high capacity and highly reversible LiNiO2 material without metal substitutions and without increasing the charge cut-off voltage.

  20. Development of Site-Specific Soil Design Basis Earthquake (DBE) Parameters for the Integrated Waste Treatment Unit (IWTU)

    SciTech Connect

    Payne, Suzette

    2008-08-01

    Horizontal and vertical PC 3 (2,500 yr) Soil Design Basis Earthquake (DBE) 5% damped spectra, corresponding time histories, and strain-compatible soil properties were developed for the Integrated Waste Treatment Unit (IWTU). The IWTU is located at the Idaho Nuclear Technology and Engineering Center (INTEC) at the Idaho National Laboratory (INL). Mean and 84th percentile horizontal DBE spectra derived from site-specific site response analyses were evaluated for the IWTU. The horizontal and vertical PC 3 (2,500 yr) Soil DBE 5% damped spectra at the 84th percentile were selected for Soil Structure Interaction (SSI) analyses at IWTU. The site response analyses were performed consistent with applicable Department of Energy (DOE) Standards, recommended guidance of the Nuclear Regulatory Commission (NRC), American Society of Civil Engineers (ASCE) Standards, and recommendations of the Blue Ribbon Panel (BRP) and Defense Nuclear Facilities Safety Board (DNFSB).

  1. Design of protease-resistant myelin basic protein-derived peptides by cleavage site directed amino acid substitutions.

    PubMed

    Burster, Timo; Marin-Esteban, Viviana; Boehm, Bernhard O; Dunn, Shannon; Rotzschke, Olaf; Falk, Kirsten; Weber, Ekkehard; Verhelst, Steven H L; Kalbacher, Hubert; Driessen, Christoph

    2007-11-15

    Multiple Sclerosis (MS) is considered to be a T cell-mediated autoimmune disease. An attractive strategy to prevent activation of autoaggressive T cells in MS, is the use of altered peptide ligands (APL), which bind to major histocompatibility complex class II (MHC II) molecules. To be of clinical use, APL must be capable of resisting hostile environments including the proteolytic machinery of antigen presenting cells (APC). The current design of APL relies on cost- and labour-intensive strategies. To overcome these major drawbacks, we used a deductive approach which involved modifying proteolytic cleavage sites in APL. Cleavage site-directed amino acid substitution of the autoantigen myelin basic protein (MBP) resulted in lysosomal protease-resistant, high-affinity binding peptides. In addition, these peptides mitigated T cell activation in a similar fashion as conventional APL. The strategy outlined allows the development of protease-resistant APL and provides a universal design strategy to improve peptide-based immunotherapeutics.

  2. Teaching E-Commerce Web Page Evaluation and Design: A Pilot Study Using Tourism Destination Sites

    ERIC Educational Resources Information Center

    Susser, Bernard; Ariga, Taeko

    2006-01-01

    This study explores a teaching method for improving business students' skills in e-commerce page evaluation and making Web design majors aware of business content issues through cooperative learning. Two groups of female students at a Japanese university studying either tourism or Web page design were assigned tasks that required cooperation to…

  3. Teaching E-Commerce Web Page Evaluation and Design: A Pilot Study Using Tourism Destination Sites

    ERIC Educational Resources Information Center

    Susser, Bernard; Ariga, Taeko

    2006-01-01

    This study explores a teaching method for improving business students' skills in e-commerce page evaluation and making Web design majors aware of business content issues through cooperative learning. Two groups of female students at a Japanese university studying either tourism or Web page design were assigned tasks that required cooperation to…

  4. Design & Technology and the Historic Environment: A Teacher's Guide. Education on Site.

    ERIC Educational Resources Information Center

    Barnes, Jonathan

    Design and technology are subjects that are about the real world, involving the application of scientific and related knowledge to a problem. The built environment is an excellent context for work in this area, as every building involves both design ideas and technological skills. This guide suggests practical activities and ideas for using the…

  5. Two Simple Leg Net Devices Designed to Protect Lower-Extremity Skin Grafts and Donor Sites and Prevent Decubitus Ulcer

    DTIC Science & Technology

    2007-02-01

    Two Simple Leg Net Devices Designed to Protect Lower-Extremity Skin Grafts and Donor Sites and Prevent Decubitus Ulcer Travis L. Hedman, MPT, OCS... decubitus . Pressure ulcer is a serious health prob- lem and can cause pain, suffering, disability, and even death.1,2 The cost of treatment for a...single pressure decubitus has been estimated to be as high as $70,000.3 Therefore, prevention is paramount. The prevention of pressure ulcers is far less

  6. Sampling design and procedures for fixed surface-water sites in the Georgia-Florida coastal plain study unit, 1993

    USGS Publications Warehouse

    Hatzell, H.H.; Oaksford, E.T.; Asbury, C.E.

    1995-01-01

    The implementation of design guidelines for the National Water-Quality Assessment (NAWQA) Program has resulted in the development of new sampling procedures and the modification of existing procedures commonly used in the Water Resources Division of the U.S. Geological Survey. The Georgia-Florida Coastal Plain (GAFL) study unit began the intensive data collection phase of the program in October 1992. This report documents the implementation of the NAWQA guidelines by describing the sampling design and procedures for collecting surface-water samples in the GAFL study unit in 1993. This documentation is provided for agencies that use water-quality data and for future study units that will be entering the intensive phase of data collection. The sampling design is intended to account for large- and small-scale spatial variations, and temporal variations in water quality for the study area. Nine fixed sites were selected in drainage basins of different sizes and different land-use characteristics located in different land-resource provinces. Each of the nine fixed sites was sampled regularly for a combination of six constituent groups composed of physical and chemical constituents: field measurements, major ions and metals, nutrients, organic carbon, pesticides, and suspended sediments. Some sites were also sampled during high-flow conditions and storm events. Discussion of the sampling procedure is divided into three phases: sample collection, sample splitting, and sample processing. A cone splitter was used to split water samples for the analysis of the sampling constituent groups except organic carbon from approximately nine liters of stream water collected at four fixed sites that were sampled intensively. An example of the sample splitting schemes designed to provide the sample volumes required for each sample constituent group is described in detail. Information about onsite sample processing has been organized into a flowchart that describes a pathway for each of

  7. Developing Educational Materials about Risks on Social Network Sites: A Design Based Research Approach

    ERIC Educational Resources Information Center

    Vanderhoven, Ellen; Schellens, Tammy; Vanderlinde, Ruben; Valcke, Martin

    2016-01-01

    Nearly all of today's Western teenagers have a profile on a social network site (SNS). As many risks have been reported, researchers and governments have emphasized the role of school education to teach teenagers how to deal safely with SNSs. However, little is known about the specific characteristics which would make interventions effective.…

  8. 10 CFR 61.51 - Disposal site design for land disposal.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... extent practicable water infiltration, to direct percolating or surface water away from the disposed... must direct surface water drainage away from disposal units at velocities and gradients which will not result in erosion that will require ongoing active maintenance in the future. (6) The disposal site...

  9. Developing Educational Materials about Risks on Social Network Sites: A Design Based Research Approach

    ERIC Educational Resources Information Center

    Vanderhoven, Ellen; Schellens, Tammy; Vanderlinde, Ruben; Valcke, Martin

    2016-01-01

    Nearly all of today's Western teenagers have a profile on a social network site (SNS). As many risks have been reported, researchers and governments have emphasized the role of school education to teach teenagers how to deal safely with SNSs. However, little is known about the specific characteristics which would make interventions effective.…

  10. Software Design for Wireless Sensor-based Site-specific Irrigation

    USDA-ARS?s Scientific Manuscript database

    In-field sensor-based site-specific irrigation management is of benefit to producers for efficient water management. Integration of the decision making process with the controls is a viable option for determining when and where to irrigate, and how much water to apply. This research presents the des...

  11. Design, development and evaluation of a tree planting-site-specific fumigant applicator

    USDA-ARS?s Scientific Manuscript database

    The goal of this research was to use recent advances in the global positioning system (GPS) and computer technology to apply just the right amount of fumigant where it is most needed (i.e., tree-planting-site-specific application) to decrease the incidence of replant disease, and achieve the environ...

  12. A rationally designed fluorescence chemosensor for on-site monitoring of carbon monoxide in air.

    PubMed

    Yan, Tengfei; Chen, Jin; Wu, Song; Mao, Zhiqiang; Liu, Zhihong

    2014-06-20

    A fluorescence chemsensor for carbon monoxide (CO), based on transformation of weakly fluorescent iodide to strong fluorescent amino product upon reacting with CO, shows abilities of quantitative measurement of CO in air at a level of 50-1000 ppm and real-time and on-site monitoring for CO flammation/explosion.

  13. Effects of sampling design on age ratios of migrants captured at stopover sites

    Treesearch

    Jeffrey F. Kelly; Deborah M. Finch

    2000-01-01

    Age classes of migrant songbirds often differ in migration timing. This difference creates the potential for age-ratios recorded at stopover sites to vary with the amount and distribution of sampling effort used. To test for these biases, we sub-sampled migrant capture data from the Middle Rio Grande Valley of New Mexico. We created data sets that reflected the age...

  14. Environmental design criteria for the 1/3 scale OTEC cold water pipe at-sea test site off Honolulu, Hawaii

    NASA Astrophysics Data System (ADS)

    1983-01-01

    In order to expedite development of the design of the 1/3 scale At-Sea Test CWP/Platform/Mooring System, environmental design criteria data at the proposed At-Sea Test site evaluated from available historic data is provided. An evaluation of the available data to date is described. It is expected that as the design develops actual site specific observation will be obtained to verify these critical design criteria.

  15. Computational design of an endo-1,4-[beta]-xylanase ligand binding site

    SciTech Connect

    Morin, Andrew; Kaufmann, Kristian W.; Fortenberry, Carie; Harp, Joel M.; Mizoue, Laura S.; Meiler, Jens

    2012-09-05

    The field of computational protein design has experienced important recent success. However, the de novo computational design of high-affinity protein-ligand interfaces is still largely an open challenge. Using the Rosetta program, we attempted the in silico design of a high-affinity protein interface to a small peptide ligand. We chose the thermophilic endo-1,4-{beta}-xylanase from Nonomuraea flexuosa as the protein scaffold on which to perform our designs. Over the course of the study, 12 proteins derived from this scaffold were produced and assayed for binding to the target ligand. Unfortunately, none of the designed proteins displayed evidence of high-affinity binding. Structural characterization of four designed proteins revealed that although the predicted structure of the protein model was highly accurate, this structural accuracy did not translate into accurate prediction of binding affinity. Crystallographic analyses indicate that the lack of binding affinity is possibly due to unaccounted for protein dynamics in the 'thumb' region of our design scaffold intrinsic to the family 11 {beta}-xylanase fold. Further computational analysis revealed two specific, single amino acid substitutions responsible for an observed change in backbone conformation, and decreased dynamic stability of the catalytic cleft. These findings offer new insight into the dynamic and structural determinants of the {beta}-xylanase proteins.

  16. Computational design of an endo-1,4-β-xylanase ligand binding site

    PubMed Central

    Morin, Andrew; Kaufmann, Kristian W.; Fortenberry, Carie; Harp, Joel M.; Mizoue, Laura S.; Meiler, Jens

    2011-01-01

    The field of computational protein design has experienced important recent success. However, the de novo computational design of high-affinity protein–ligand interfaces is still largely an open challenge. Using the Rosetta program, we attempted the in silico design of a high-affinity protein interface to a small peptide ligand. We chose the thermophilic endo-1,4-β-xylanase from Nonomuraea flexuosa as the protein scaffold on which to perform our designs. Over the course of the study, 12 proteins derived from this scaffold were produced and assayed for binding to the target ligand. Unfortunately, none of the designed proteins displayed evidence of high-affinity binding. Structural characterization of four designed proteins revealed that although the predicted structure of the protein model was highly accurate, this structural accuracy did not translate into accurate prediction of binding affinity. Crystallographic analyses indicate that the lack of binding affinity is possibly due to unaccounted for protein dynamics in the ‘thumb’ region of our design scaffold intrinsic to the family 11 β-xylanase fold. Further computational analysis revealed two specific, single amino acid substitutions responsible for an observed change in backbone conformation, and decreased dynamic stability of the catalytic cleft. These findings offer new insight into the dynamic and structural determinants of the β-xylanase proteins. PMID:21349882

  17. Remedial Action Plan and site design for stabilization of the inactive uranium mill tailings site at Gunnison, Colorado. Attachment 5, Supplemental radiological data: Final report

    SciTech Connect

    Not Available

    1992-10-01

    Diffusion coefficients for radon gas in earthen materials are required to design suitable radon-barrier covers for uranium tailings impoundments and other materials that emit radon gas. Many early measurements of radon diffusion coefficients relied on the differences in steady-state radon fluxes measured from radon source before and after installation of a cover layer of the material being tested. More recent measurements have utilized the small-sample transient (SST) technique for greater control on moistures and densities of the test soils, greater measurement precision, and reduced testing time and costs. Several of the project sites for the US Department of Energy`s Uranium Mill Tailings Remedial Action (UMTRA) Program contain radiologically contaminated subsurface material composed predominantly of cobbles, gravels andsands. Since remedial action designs require radon diffusion coefficients for the source materials as well as the cover materials, these cobbly and gravelly materials also must be tested. This report contains the following information: a description of the test materials used and the methods developed to conduct the SST radon diffusion measurements on cobbly soils; the protocol for conducting radon diffusion tests oncobbly soils; the results of measurements on the test samples; and modifications to the FITS computer code for analyzing the time-dependent radon diffusion data.

  18. Environmental monitoring at designed geopressured-geothermal well sites, Louisiana and Texas

    SciTech Connect

    Not Available

    1990-01-01

    This document covers the activities of monitoring environmental aspects at designated geothermal wells in Texas and Louisiana during the second quarter of 1990 by the Louisiana Geological Survey, Louisiana State University under contract with US DOE. 1 fig. (FSD)

  19. Use of technical and economic analysis for optimizing technology selection and remedial design for contaminated sites

    SciTech Connect

    Hardisty, P.E.; Brown, A.

    1996-12-01

    The decision to remediate a contaminated site can be seen from the macroeconomic and microeconomic viewpoints. Macroeconomics can be used to plan and account for the overall cost of pollution as part of a firm`s production, and thus make overall decisions on the real cost of pollution and the level of clean-up which may be called for. Valuation of damaged resources, option values and intrinsic worth is an important part of this process. Once the decision to remediate has been taken, the question becomes how best to remediate. Microeconomic analysis deals with providing efficient allocative decisions for reaching specified goals. it is safe to say that cost is one of the single most important factors in site clean-up decision making. A basic rule of remediation is often taken to be the maximization of contaminant mass removed per dollar spent. However, remediation may also be governed by other objectives and constraints. In some situations, minimization of time, rather than cost, could be the constraint. Or perhaps the objective could be to achieve a set level of clean-up for the lowest possible cost, even if a large program would result in unit-cost reductions. Evaluation of the economics of a clean-up project is directly linked to the objectives of the site owner, and the constraints within which the remediation is to be performed. Economic analysis of remedial options for containment of a 350,000 L hydrocarbon spill migrating through fractured rock into a river in Alberta, Canada, clear direction to the site owner.

  20. Synthetic design of heterometallic cluster compounds with site-selective and stepwise substitution of bridging carboxylates.

    PubMed

    Ikegami, Atsushi; Abe, Masaaki; Inatomi, Atsushi; Hisaeda, Yoshio

    2010-04-19

    At the right places: Owing to substitution-lability difference in disparate metal centers, an oxo-centered heterometallic trinuclear acetato complex [Ru2MgO(CH3COO)6(py)3] (py = pyridine; see picture) undergoes site-selective substitution of propionates and ferrocenycarboxylates in a stepwise manner. The new substituted families have been successfully separated by chromatography and fully characterized. Copyright © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Remedial Action Plan and Site Design for stabilization of the inactive Uranium Mill Tailings sites at Slick Rock, Colorado: Appendix C to Attachment 3, Calculations. Final

    SciTech Connect

    1995-09-01

    This volume contains calculations for: Slick Rock processing sites background ground water quality; Slick Rock processing sites lysimeter water quality; Slick Rock processing sites on-site and downgradient ground water quality; Slick Rock disposal site background water quality; Burro Canyon disposal site, Slick Rock, Colorado, average hydraulic gradients and average liner ground water velocities in the upper, middle, and lower sandstone units of the Burro Canyon formation; Slick Rock--Burro Canyon disposal site, Burro Canyon pumping and slug tests--analyses; water balance and surface contours--Burro Canyon disposal cell; and analytical calculation of drawdown in a hypothetical well completed in the upper sandstone unit of the Burro Canyon formation.

  2. Remedial action and site design for stabilization of the inactive uranium mill tailings sites at Slick Rock, Colorado. Attachment 3, Groundwater hydrology report

    SciTech Connect

    Not Available

    1993-07-01

    The US Environmental Protection Agency (EPA) has established health and environmental protection regulations to correct and prevent groundwater contamination resulting from processing activities at inactive uranium milling sites (EPA, 1987). According to the Uranium Mill Tailings Radiation Control Act (UMTRCA) of 1978 Public Law (PL) 95-604 (PL 95-604), the US Department of Energy (DOE) is responsible for assessing the inactive uranium processing sites. The DOE has determined that for Slick Rock, this assessment shall include hydrogeologic site characterization for two separate uranium processing sites, the Union Carbide (UC) site and the North Continent (NC) site, and for the proposed Burro Canyon disposal site.

  3. Environmental Aspects of Two Volatile Organic Compound Groundwater Treatment Designs at the Rocky Flats Site - 13135

    SciTech Connect

    Michalski, Casey C.; DiSalvo, Rick; Boylan, John

    2013-07-01

    DOE's Rocky Flats Site in Colorado is a former nuclear weapons production facility that began operations in the early 1950's. Because of releases of hazardous substances to the environment, the federally owned property and adjacent offsite areas were placed on the CERCLA National Priorities List in 1989. The final remedy was selected in 2006. Engineered components of the remedy include four groundwater treatment systems that were installed before closure as CERCLA-accelerated actions. Two of the systems, the Mound Site Plume Treatment System and the East Trenches Plume Treatment System, remove low levels of volatile organic compounds using zero-valent iron media, thereby reducing the loading of volatile organic compounds in surface water resulting from the groundwater pathway. However, the zero-valent iron treatment does not reliably reduce all volatile organic compounds to consistently meet water quality goals. While adding additional zero-valent iron media capacity could improve volatile organic compound removal capability, installation of a solar powered air-stripper has proven an effective treatment optimization in further reducing volatile organic compound concentrations. A comparison of the air stripper to the alternative of adding additional zero-valent iron capacity to improve Mound Site Plume Treatment System and East Trenches Plume Treatment System treatment based on several key sustainable remediation aspects indicates the air stripper is also more 'environmentally friendly'. These key aspects include air pollutant emissions, water quality, waste management, transportation, and costs. (authors)

  4. Demonstration of the importance and usefulness of manipulating non-active-site residues in protein design.

    PubMed

    Shimotohno, A; Oue, S; Yano, T; Kuramitsu, S; Kagamiyama, H

    2001-06-01

    Do non-active-site residues participate in protein function in a more direct way than just by holding the static framework of the protein molecule? If so, how important are they? As a model to answer these questions, ATB17, which is a mutant of aspartate aminotransferase created by directed evolution, is an ideal system because it shows a 10(6)-fold increase in the catalytic efficiency for valine but most of its 17 mutated residues are non-active-site residues. To analyze the roles of the mutations in the altered function, we divided the mutations into four groups, namely, three clusters and the remainder, based on their locations in the three-dimensional structure. Mutants with various combinations of the clusters were constructed and analyzed, and the data were interpreted in the context of the structure-function relationship of this enzyme. Each cluster shows characteristic effects: for example, one cluster appears to enhance the catalytic efficiency by fixing the conformation of the enzyme to that of the substrate-bound form. The effects of the clusters are largely additive and independent of each other. The present results illustrate how a protein function is dramatically modified by the accumulation of many seemingly inert mutations of non-active-site residues.

  5. "Less Clicking, More Watching": Results from the User-Centered Design of a Multi-Institutional Web Site for Art and Culture.

    ERIC Educational Resources Information Center

    Vergo, John; Karat, Clare-Marie; Karat, John; Pinhanez, Claudio; Arora, Renee; Cofino, Thomas; Riecken, Doug; Podlaseck, Mark

    This paper summarizes a 10-month long research project conducted at the IBM T.J. Watson Research Center aimed at developing the design concept of a multi-institutional art and culture web site. The work followed a user-centered design (UCD) approach, where interaction with prototypes and feedback from potential users of the web site were sought…

  6. HPC Analysis of Multiple Binding Sites Communication and Allosteric Modulations in Drug Design: The HSP Case Study.

    PubMed

    Chiappori, Federica; Milanesi, Luciano; Merelli, Ivan

    2016-01-01

    Allostery is a long-range macromolecular mechanism of internal regulation, in which the binding of a ligand in an allosteric site induces distant conformational changes in a distant portion of the protein, modifying its activity. From the drug design point of view, this mechanism can be exploited to achieve important therapeutic effects, since ligands able to bind allosteric sites may be designed to regulate target proteins. Computational tools are a valid support in this sense, since they allow the characterization of allosteric communications within proteins, which are essential to design modulator ligands. While considering long-range interactions in macromolecules, the principal drug design tool available to researcher is molecular dynamics, and related applications, since it allows the evaluation of conformational changes of a protein bound to a ligand. In particular, all-atoms molecular dynamics is suitable to verify the internal mechanisms that orchestrate allosteric communications, in order to identify key residues and internal pathways that modify the protein behaviour. The problem is that these techniques are heavily time-consuming and computationally intensive, thus high performance computing systems, including parallel computing and GPU-accelerated computations, are necessary to achieve results in a reasonable time. In this review, we will discuss how it is possible to exploit in silico approaches to characterize allosteric modulations and long-range interactions within proteins, describing the case study of the Heat Shock Proteins, a class of chaperons regulated by stress conditions, which is particularly important since it is involved in many cancers and neurodegenerative diseases.

  7. Design.

    ERIC Educational Resources Information Center

    Online-Offline, 1998

    1998-01-01

    Provides an annotated bibliography of resources on this month's theme "Design" for K-8 language arts, art and architecture, music and dance, science, math, social studies, health, and physical education. Includes Web sites, CD-ROMs and software, videos, books, audiotapes, magazines, professional resources and classroom activities.…

  8. Bayesian Geostatistical Design: Optimal Site Investigation When the Geostatistical Model is Uncertain

    NASA Astrophysics Data System (ADS)

    Nowak, W.; de Barros, F. P. J.; Rubin, Y.

    2009-04-01

    Geostatistical optimal design optimizes subsurface exploration for maximum information towards task-specific prediction goals. Until recently, geostatistical design studies have assumed that the geostatistical description (i.e., the mean, trends, covariance models and their parameters) is given a priori, even if only few or no data offer support for such assumptions. This is in contradiction with the fact that the bulk of data acquisition is merely being planned at this stage. We believe that geostatistical design should comply with the following four guidelines: 1. Avoid unjustified a priori assumptions on the geostatistical description such as claiming certainty in the geostatistical model, but to acknowledge the inevitable uncertainty of geostatistical descriptions, 2. Reduce geostatistical model uncertainty as secondary design objective, 3. Rate this secondary objective optimal for the overall prediction goal and 4. Be robust even under inaccurate geostatistical assumptions. Bayesian Geostatistical Design (Diggle und Lophaven, 2006) follows the above four guidelines by considering uncertain covariance model parameters. These authors considered a kriging-like prediction task, using the spatial average of the estimation variance as objective function for the design. We transfer their concept from kriging-like applications to geostatistical inverse problems, thus generalizing towards arbitrary hydrogeological or geophysical data and prediction goals. A remaining concern is that we deem it inappropriate to consider parametric uncertainty only within a single covariance model. The Matérn family of covariance functions has an additional shape parameter, and so allows for uncertain smoothness and shape of the covariance function (Zhang and Rubin, submitted to WRR). Controlling model shape by a parameter converts covariance model selection to parameter identification and resembles Bayesian model averaging over a continuous spectrum of covariance models. We illustrate

  9. Engineered materials characterization report for the Yucca Mountain Site Characterization Project. Volume 2, Design data

    SciTech Connect

    Konynenburg, R.A.; McCright, R.D.; Roy, A.K.; Jones, D.A.

    1995-08-01

    This is Volume 2 of the Engineered Materials Characterization Report which presents the design data for candidate materials needed in fabricating different components for both large and medium multi-purpose canister (MPC) disposal containers, waste packages for containing uncanistered spent fuel (UCF), and defense high-level waste (HLW) glass disposal containers. The UCF waste package consists of a disposal container with a basket therein. It is assumed that the waste packages will incorporate all-metallic multibarrier disposal containers to accommodate medium and large MPCs, ULCF, and HLW glass canisters. Unless otherwise specified, the disposal container designs incorporate an outer corrosion-allowance metal barrier over an inner corrosion-resistant metal barrier. The corrosion-allowance barrier, which will be thicker than the inner corrosion-resistant barrier, is designed to undergo corrosion-induced degradation at a very low rate, thus providing the inner barrier protection from the near-field environment for a prolonged service period.

  10. Biological Characterization of an Improved Pyrrole-Based Colchicine Site Agent Identified through Structure-Based Design

    PubMed Central

    Rohena, Cristina C.; Telang, Nakul S.; Da, Chenxiao; Risinger, April L.; Sikorski, James A.; Kellogg, Glen E.; Gupton, John T.

    2016-01-01

    A refined model of the colchicine site on tubulin was used to design an improved analog of the pyrrole parent compound, JG-03-14. The optimized compound, NT-7-16, was evaluated in biological assays that confirm that it has potent activities as a new colchicine site microtubule depolymerizer. NT-7-16 exhibits antiproliferative and cytotoxic activities against multiple cancer cell lines, with IC50 values of 10–16 nM, and it is able to overcome drug resistance mediated by the expression of P-glycoprotein and the βIII isotype of tubulin. NT-7-16 initiated the concentration-dependent loss of cellular microtubules and caused the formation of abnormal mitotic spindles, leading to mitotic accumulation. The direct interaction of NT-7-16 with purified tubulin was confirmed, and it was more potent than combretastatin A-4 in these assays. Binding studies verified that NT-7-16 binds to tubulin within the colchicine site. The antitumor effects of NT-7-16 were evaluated in an MDA-MB-435 xenograft model and it had excellent activity at concentrations that were not toxic. A second compound, NT-9-21, which contains dichloro moieties in place of the 3,5-dibromo substituents of NT-7-16, had a poorer fit within the colchicine site as predicted by modeling and the Hydropathic INTeractions score. Biological evaluations showed that NT-9-21 has 10-fold lower potency than NT-7-16, confirming the modeling predictions. These studies highlight the value of the refined colchicine-site model and identify a new pyrrole-based colchicine-site agent with potent in vitro activities and promising in vivo antitumor actions. PMID:26655304

  11. Fly ash design manual for road and site applications. Volume 1. Dry or conditioned placement

    SciTech Connect

    DiGioia, A.M.; McLaren, R.J.; Burns, D.L.; Miller, D.E.

    1986-02-01

    This updated design manual describes the use of fly ash as a construction material for use as structural and nonstructural fills, backfills, embankments, base courses, soil stabilization, land reclamation and other high volume uses. The manual details the physical, engineering, and chemical properties of bituminous, subbituminous and lignite fly ash. Included are field and laboratory testing methods, design data, procedures and examples, specifications, quality control, and pre- and post-construction monitoring. Volume 1 describes uses where fly ash is used dry or conditioned with small amounts of moisture. Volume 2 describes uses where fly ash is placed as a slurry with relatively large amounts of water.

  12. The Phylogeny and Active Site Design of Eukaryotic Copper-only Superoxide Dismutases

    SciTech Connect

    Peterson, Ryan L.; Galaleldeen, Ahmad; Villarreal, Johanna; Taylor, Alexander B.; Cabelli, Diane E.; Hart, P. John; Culotta, Valeria C.

    2016-08-17

    In eukaryotes the bimetallic Cu/Zn superoxide dismutase (SOD) enzymes play important roles in the biology of reactive oxygen species by disproportionating superoxide anion. We reported that the fungal pathogen Candida albicans expresses a novel copper-only SOD, known as SOD5, that lacks the zinc cofactor and electrostatic loop (ESL) domain of Cu/Zn-SODs for substrate guidance. In spite of these abnormalities, C. albicans SOD5 can disproportionate superoxide at rates limited only by diffusion. Here we demonstrate that this curious copper-only SOD occurs throughout the fungal kingdom as well as in phylogenetically distant oomycetes or “pseudofungi” species. It is the only form of extracellular SOD in fungi and oomycetes, in stark contrast to the extracellular Cu/Zn-SODs of plants and animals. Through structural biology and biochemical approaches we demonstrate that these copper-only SODs have evolved with a specialized active site consisting of two highly conserved residues equivalent to SOD5 Glu-110 and Asp-113. The equivalent positions are zinc binding ligands in Cu/Zn-SODs and have evolved in copper-only SODs to control catalysis and copper binding in lieu of zinc and the ESL. Similar to the zinc ion in Cu/Zn-SODs, SOD5 Glu-110 helps orient a key copper-coordinating histidine and extends the pH range of enzyme catalysis. Furthermore, SOD5 Asp-113 connects to the active site in a manner similar to that of the ESL in Cu/Zn-SODs and assists in copper cofactor binding. Copper-only SODs are virulence factors for certain fungal pathogens; thus this unique active site may be a target for future anti-fungal strategies.

  13. The Phylogeny and Active Site Design of Eukaryotic Copper-only Superoxide Dismutases.

    PubMed

    Peterson, Ryan L; Galaleldeen, Ahmad; Villarreal, Johanna; Taylor, Alexander B; Cabelli, Diane E; Hart, P John; Culotta, Valeria C

    2016-09-30

    In eukaryotes the bimetallic Cu/Zn superoxide dismutase (SOD) enzymes play important roles in the biology of reactive oxygen species by disproportionating superoxide anion. Recently, we reported that the fungal pathogen Candida albicans expresses a novel copper-only SOD, known as SOD5, that lacks the zinc cofactor and electrostatic loop (ESL) domain of Cu/Zn-SODs for substrate guidance. Despite these abnormalities, C. albicans SOD5 can disproportionate superoxide at rates limited only by diffusion. Here we demonstrate that this curious copper-only SOD occurs throughout the fungal kingdom as well as in phylogenetically distant oomycetes or "pseudofungi" species. It is the only form of extracellular SOD in fungi and oomycetes, in stark contrast to the extracellular Cu/Zn-SODs of plants and animals. Through structural biology and biochemical approaches we demonstrate that these copper-only SODs have evolved with a specialized active site consisting of two highly conserved residues equivalent to SOD5 Glu-110 and Asp-113. The equivalent positions are zinc binding ligands in Cu/Zn-SODs and have evolved in copper-only SODs to control catalysis and copper binding in lieu of zinc and the ESL. Similar to the zinc ion in Cu/Zn-SODs, SOD5 Glu-110 helps orient a key copper-coordinating histidine and extends the pH range of enzyme catalysis. SOD5 Asp-113 connects to the active site in a manner similar to that of the ESL in Cu/Zn-SODs and assists in copper cofactor binding. Copper-only SODs are virulence factors for certain fungal pathogens; thus this unique active site may be a target for future anti-fungal strategies. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  14. The Screening India's Twin Epidemic: Study design and methodology (SITE-1).

    PubMed

    Joshi, Shashank R; Vadivale, Muruga; Dalal, Jamshed J; Das, Ashok Kumar

    2011-10-01

    The recent years have seen a surge in the prevalence of both diabetes and hypertension. Significant demographic variations reported on the prevalence patterns of diabetes and hypertension in India establish a clear need for a nation-wide surveillance study. The Screening India's Twin Epidemic (SITE) study aimed at collecting information on the prevalence of diagnosed and undiagnosed diabetes and hypertension cases in outpatient settings in major Indian states to better understand disease management, as well as to estimate the extent of underlying risk factors. During 2009-2010, SITE was conducted in eight states, in waves - one state at a time. It was planned to recruit about 2000 patients from 100 centers per wave. Each center enrolled the first 10 eligible patients (≥18 years of age, not pregnant, signed data release consent form, and ready to undergo screening tests) per day on two consecutive days. Patient demographics, medical history, and laboratory investigation results were collected and statistically interpreted. The protocol defined diabetes and hypertension as per the American Diabetes Association (ADA) and Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC 7) recommendations, respectively. After the first two pilot phases in Maharashtra and Delhi, the protocol was refined and the laboratory investigations were simplified to be further employed for all other states, namely, Tamil Nadu, West Bengal, Karnataka, Andhra Pradesh, Madhya Pradesh, and Gujarat. SITE's nation-wide approach will provide a real-world perspective on diabetes and hypertension and its contributing risk factors. Results from the study will raise awareness on the need for early diagnosis and management of these diseases to reduce complications.

  15. The Screening India's Twin Epidemic: Study design and methodology (SITE-1)

    PubMed Central

    Joshi, Shashank R.; Vadivale, Muruga; Dalal, Jamshed J.; Das, Ashok Kumar

    2011-01-01

    Objectives: The recent years have seen a surge in the prevalence of both diabetes and hypertension. Significant demographic variations reported on the prevalence patterns of diabetes and hypertension in India establish a clear need for a nation-wide surveillance study. The Screening India's Twin Epidemic (SITE) study aimed at collecting information on the prevalence of diagnosed and undiagnosed diabetes and hypertension cases in outpatient settings in major Indian states to better understand disease management, as well as to estimate the extent of underlying risk factors. Materials and Methods: During 2009–2010, SITE was conducted in eight states, in waves – one state at a time. It was planned to recruit about 2000 patients from 100 centers per wave. Each center enrolled the first 10 eligible patients (≥18 years of age, not pregnant, signed data release consent form, and ready to undergo screening tests) per day on two consecutive days. Patient demographics, medical history, and laboratory investigation results were collected and statistically interpreted. The protocol defined diabetes and hypertension as per the American Diabetes Association (ADA) and Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC 7) recommendations, respectively. Results: After the first two pilot phases in Maharashtra and Delhi, the protocol was refined and the laboratory investigations were simplified to be further employed for all other states, namely, Tamil Nadu, West Bengal, Karnataka, Andhra Pradesh, Madhya Pradesh, and Gujarat. Conclusion: SITE's nation-wide approach will provide a real-world perspective on diabetes and hypertension and its contributing risk factors. Results from the study will raise awareness on the need for early diagnosis and management of these diseases to reduce complications. PMID:22145145

  16. The Phylogeny and Active Site Design of Eukaryotic Copper-only Superoxide Dismutases

    SciTech Connect

    Peterson, Ryan L.; Galaleldeen, Ahmad; Villarreal, Johanna; Taylor, Alexander B.; Cabelli, Diane E.; Hart, P. John; Culotta, Valeria C.

    2016-08-17

    In eukaryotes the bimetallic Cu/Zn superoxide dismutase (SOD) enzymes play important roles in the biology of reactive oxygen species by disproportionating superoxide anion. We reported that the fungal pathogen Candida albicans expresses a novel copper-only SOD, known as SOD5, that lacks the zinc cofactor and electrostatic loop (ESL) domain of Cu/Zn-SODs for substrate guidance. In spite of these abnormalities, C. albicans SOD5 can disproportionate superoxide at rates limited only by diffusion. Here we demonstrate that this curious copper-only SOD occurs throughout the fungal kingdom as well as in phylogenetically distant oomycetes or “pseudofungi” species. It is the only form of extracellular SOD in fungi and oomycetes, in stark contrast to the extracellular Cu/Zn-SODs of plants and animals. Through structural biology and biochemical approaches we demonstrate that these copper-only SODs have evolved with a specialized active site consisting of two highly conserved residues equivalent to SOD5 Glu-110 and Asp-113. The equivalent positions are zinc binding ligands in Cu/Zn-SODs and have evolved in copper-only SODs to control catalysis and copper binding in lieu of zinc and the ESL. Similar to the zinc ion in Cu/Zn-SODs, SOD5 Glu-110 helps orient a key copper-coordinating histidine and extends the pH range of enzyme catalysis. Furthermore, SOD5 Asp-113 connects to the active site in a manner similar to that of the ESL in Cu/Zn-SODs and assists in copper cofactor binding. Copper-only SODs are virulence factors for certain fungal pathogens; thus this unique active site may be a target for future anti-fungal strategies.

  17. The Phylogeny and Active Site Design of Eukaryotic Copper-only Superoxide Dismutases

    DOE PAGES

    Peterson, Ryan L.; Galaleldeen, Ahmad; Villarreal, Johanna; ...

    2016-08-17

    In eukaryotes the bimetallic Cu/Zn superoxide dismutase (SOD) enzymes play important roles in the biology of reactive oxygen species by disproportionating superoxide anion. We reported that the fungal pathogen Candida albicans expresses a novel copper-only SOD, known as SOD5, that lacks the zinc cofactor and electrostatic loop (ESL) domain of Cu/Zn-SODs for substrate guidance. In spite of these abnormalities, C. albicans SOD5 can disproportionate superoxide at rates limited only by diffusion. Here we demonstrate that this curious copper-only SOD occurs throughout the fungal kingdom as well as in phylogenetically distant oomycetes or “pseudofungi” species. It is the only form ofmore » extracellular SOD in fungi and oomycetes, in stark contrast to the extracellular Cu/Zn-SODs of plants and animals. Through structural biology and biochemical approaches we demonstrate that these copper-only SODs have evolved with a specialized active site consisting of two highly conserved residues equivalent to SOD5 Glu-110 and Asp-113. The equivalent positions are zinc binding ligands in Cu/Zn-SODs and have evolved in copper-only SODs to control catalysis and copper binding in lieu of zinc and the ESL. Similar to the zinc ion in Cu/Zn-SODs, SOD5 Glu-110 helps orient a key copper-coordinating histidine and extends the pH range of enzyme catalysis. Furthermore, SOD5 Asp-113 connects to the active site in a manner similar to that of the ESL in Cu/Zn-SODs and assists in copper cofactor binding. Copper-only SODs are virulence factors for certain fungal pathogens; thus this unique active site may be a target for future anti-fungal strategies.« less

  18. Seismic design spectra 200 West and East Areas DOE Hanford Site, Washington

    SciTech Connect

    Tallman, A.M.

    1995-12-31

    This document presents equal hazard response spectra for the W236A project for the 200 East and West new high-level waste tanks. The hazard level is based upon WHC-SD-W236A-TI-002, Probabilistic Seismic Hazard Analysis, DOE Hanford Site, Washington. Spectral acceleration amplification is plotted with frequency (Hz) for horizontal and vertical motion and attached to this report. The vertical amplification is based upon the preliminary draft revision of Standard ASCE 4-86. The vertical spectral acceleration is equal to the horizontal at frequencies above 3.3Hz because of near-field, less than 15 km, sources.

  19. Development of molecular probes for the identification of extra interaction sites in the mid-gorge and peripheral sites of butyrylcholinesterase (BuChE). Rational design of novel, selective, and highly potent BuChE inhibitors.

    PubMed

    Campiani, Giuseppe; Fattorusso, Caterina; Butini, Stefania; Gaeta, Alessandra; Agnusdei, Marianna; Gemma, Sandra; Persico, Marco; Catalanotti, Bruno; Savini, Luisa; Nacci, Vito; Novellino, Ettore; Holloway, Harold W; Greig, Nigel H; Belinskaya, Tatyana; Fedorko, James M; Saxena, Ashima

    2005-03-24

    Tacrine heterobivalent ligands were designed as novel and reversible inhibitors of cholinesterases. On the basis of the investigation of the active site gorge topology of butyrylcholinesterase (BuChE) and acetylcholinesterase (AChE) and by using flexible docking procedures, molecular modeling studies formulated the hypothesis of extra interaction sites in the active gorge of hBuChE, namely, a mid-gorge interaction site and a peripheral interaction site. The design strategy led to novel BuChE inhibitors, balancing potency and selectivity. Among the compounds identified, the heterobivalent ligand 4m, containing an amide nitrogen and a sulfur atom at the 8-membered tether level, is one of the most potent and selective BuChE inhibitors described to date. The novel inhibitors, bearing postulated key features, validated the hypothesis of the presence of extra interaction sites within the hBuChE active site gorge.

  20. Evaluation of an Inverse Molecular Design Algorithm in a Model Binding Site

    PubMed Central

    Huggins, David J.; Altman, Michael D.; Tidor, Bruce

    2008-01-01

    Computational molecular design is a useful tool in modern drug discovery. Virtual screening is an approach that docks and then scores individual members of compound libraries. In contrast to this forward approach, inverse approaches construct compounds from fragments, such that the computed affinity, or a combination of relevant properties, is optimized. We have recently developed a new inverse approach to drug design based on the dead-end elimination and A* algorithms employing a physical potential function. This approach has been applied to combinatorially constructed libraries of small-molecule ligands to design high-affinity HIV-1 protease inhibitors [M. D. Altman et al. J. Am. Chem. Soc. 130: 6099–6013, 2008]. Here we have evaluated the new method using the well studied W191G mutant of cytochrome c peroxidase. This mutant possesses a charged binding pocket and has been used to evaluate other design approaches. The results show that overall the new inverse approach does an excellent job of separating binders from non-binders. For a few individual cases, scoring inaccuracies led to false positives. The majority of these involve erroneous solvation energy estimation for charged amines, anilinium ions and phenols, which has been observed previously for a variety of scoring algorithms. Interestingly, although inverse approaches are generally expected to identify some but not all binders in a library, due to limited conformational searching, these results show excellent coverage of the known binders while still showing strong discrimination of the non-binders. PMID:18831031

  1. Hybrid Placemaking in the Library: Designing Digital Technology to Enhance Users' On-Site Experience

    ERIC Educational Resources Information Center

    Bilandzic, Mark; Johnson, Daniel

    2013-01-01

    This paper presents research findings and design strategies that illustrate how digital technology can be applied as a tool for "hybrid" placemaking in ways that would not be possible in purely digital or physical spaces. Digital technology has revolutionised the way people learn and gather new information. This trend has challenged the…

  2. Making Creative Spaces: The Art and Design Classroom as a Site of Performativity

    ERIC Educational Resources Information Center

    Wild, Carol

    2011-01-01

    Rather than taking a transformational role in schools, new art and design teachers quickly become subject to "school art" orthodoxy. Theories of subjectivity and the development of professional identity within communities of practice can feel far removed from the classroom. This article seeks to make clearer the processes by which teacher identity…

  3. Evaluation of an inverse molecular design algorithm in a model binding site.

    PubMed

    Huggins, David J; Altman, Michael D; Tidor, Bruce

    2009-04-01

    Computational molecular design is a useful tool in modern drug discovery. Virtual screening is an approach that docks and then scores individual members of compound libraries. In contrast to this forward approach, inverse approaches construct compounds from fragments, such that the computed affinity, or a combination of relevant properties, is optimized. We have recently developed a new inverse approach to drug design based on the dead-end elimination and A* algorithms employing a physical potential function. This approach has been applied to combinatorially constructed libraries of small-molecule ligands to design high-affinity HIV-1 protease inhibitors (Altman et al., J Am Chem Soc 2008;130:6099-6013). Here we have evaluated the new method using the well-studied W191G mutant of cytochrome c peroxidase. This mutant possesses a charged binding pocket and has been used to evaluate other design approaches. The results show that overall the new inverse approach does an excellent job of separating binders from nonbinders. For a few individual cases, scoring inaccuracies led to false positives. The majority of these involve erroneous solvation energy estimation for charged amines, anilinium ions, and phenols, which has been observed previously for a variety of scoring algorithms. Interestingly, although inverse approaches are generally expected to identify some but not all binders in a library, due to limited conformational searching, these results show excellent coverage of the known binders while still showing strong discrimination of the nonbinders. (c) 2008 Wiley-Liss, Inc.

  4. Web-Based Learning: How Task Scaffolding and Web Site Design Support Knowledge Acquisition

    ERIC Educational Resources Information Center

    MacGregor, S. Kim; Lou, Yiping

    2005-01-01

    Using WebQuests for inquiry-based learning represents a higher-order use of technology requiring students to exercise information seeking, analyzing, and synthesizing strategies. This research was designed to obtain a better understanding of how to enhance the pedagogical effectiveness of WebQuests and of how students interact with the various…

  5. Hybrid Placemaking in the Library: Designing Digital Technology to Enhance Users' On-Site Experience

    ERIC Educational Resources Information Center

    Bilandzic, Mark; Johnson, Daniel

    2013-01-01

    This paper presents research findings and design strategies that illustrate how digital technology can be applied as a tool for "hybrid" placemaking in ways that would not be possible in purely digital or physical spaces. Digital technology has revolutionised the way people learn and gather new information. This trend has challenged the…

  6. Making Creative Spaces: The Art and Design Classroom as a Site of Performativity

    ERIC Educational Resources Information Center

    Wild, Carol

    2011-01-01

    Rather than taking a transformational role in schools, new art and design teachers quickly become subject to "school art" orthodoxy. Theories of subjectivity and the development of professional identity within communities of practice can feel far removed from the classroom. This article seeks to make clearer the processes by which teacher identity…

  7. X-ray structures of a designed binding site in trypsin show metal-dependent geometry.

    PubMed

    Brinen, L S; Willett, W S; Craik, C S; Fletterick, R J

    1996-05-14

    The three-dimensional structures of complexes of trypsin N143H, E151H bound to ecotin A86H are determined at 2.0 A resolution via X-ray crystallography in the absence and presence of the transition metals Zn2+, Ni2+, and Cu2+. The binding site for these transition metals was constructed by substitution of key amino acids with histidine at the trypsin-ecotin interface in the S2'/P2' pocket. Three histidine side chains, two on trypsin at positions 143 and 151 and one on ecotin at position 86, anchor the metals and provide extended catalytic recognition for substrates with His in the P2' pocket. Comparisons of the three-dimensional structures show the different geometries that result upon the binding of metal in the engineered tridentate site and suggest a structural basis for the kinetics of the metal-regulated catalysis. Of the three metals, the binding of zinc results in the most favorable binding geometry, not dissimilar to those observed in naturally occurring zinc binding proteins.

  8. Biomimetic Single-Site Heterogeneous Catalysts: Design Strategies and Catalytic Potential

    NASA Astrophysics Data System (ADS)

    Xuereb, David; Dzierzak, Joanna; Raja, Robert

    Enzymes catalyze the most fundamental reactions in organic chemistry from simple oxidations of straight chain alkanes to complex C-C bond forming reactions with exceptional selectivity. Mimicking the active site of an enzyme by immobilising a well defined amino acid containing transition-metal centre on a robust inorganic framework, provides a powerful catalyst that can be utilized in the production of fine chemicals and complicated drug molecules. Porous aluminosilicates and mesoporous silicas offer suitable supports for single-site bio-derived catalysts. These materials can be created from a range of methodologies and the different strategies used for immobilisation can greatly affect the nature of the active catalyst. The routes by which these catalysts are immobilised have also given the potential to derivatize inorganic structures with amino acids, not just for complexation to metal centres but for use as organocatalysts as well. These metal free bio-derivatized frameworks offer advantages over their homogeneous counterparts and can carry out stereoselective reactions with great effectiveness. Herein, the routes to heterogenizing biomimetic catalysts will be critically assessed and depending on the methods used, suitable active catalysts for use in chemo- and stereoselective transformations can be developed.

  9. Design of Visco-Elastic Dampers for RC Frame for Site-Specific Earthquake

    NASA Astrophysics Data System (ADS)

    Kamatchi, P.; Rama Raju, K.; Ravisankar, K.; Iyer, Nagesh R.

    2016-12-01

    Number of Reinforced Concrete (RC) framed buildings have got damaged at Ahmedabad city, India located at about 240 km away from epicenter during January 2001, 7.6 moment magnitude (Mw) Bhuj earthquake. In the present study, two dimensional nonlinear time history dynamic analyses of a typical 13 storey frame assumed to be located at Ahmedabad is carried out with the rock level and surface level site-specific ground motion for scenario earthquake of Mw 7.6 from Bhuj. Artificial ground motions are generated using extended finite source stochastic model with seismological parameters reported in literature for 2001 Bhuj earthquake. Surface level ground motions are obtained for a typical soil profile of 100 m depth reported in literature through one dimensional equivalent linear wave propagation analyses. From the analyses, failure of frame is observed for surface level ground motions which indicates that, in addition to the in-adequacy of the cross sections and reinforcement of the RC members of the frame chosen, the rich energy content of the surface level ground motion near the fundamental time period of the frame has also contributed for the failure of frame. As a part of retrofitting measure, five Visco-elastic Dampers (VED) in chevron bracing are added to frame. For the frame considered in the present study, provision of VED is found to be effective to mitigate damage for the soil site considered.

  10. Remedial action plan and site design for stabilization of the inactive uranium mill tailings site at Lowman, Idaho: Remedial action selection report for the Lowman UMTRA project site, Idaho

    SciTech Connect

    Matthews, M.L. . Uranium Mill Tailings Remedial Action Project Office); Nagel, J. . Div. of Environmental Quality)

    1991-09-01

    The inactive uranium mill tailings site near Lowman, Idaho, was designated as one of 24 abandoned uranium tailings sites to be remediated by the US Department of Energy (DOE) under the Uranium Mill Tailings Radiation Control Act of 1978 (UMTRCA). The UMTRCA requires that the US Nuclear Regulatory Commission (NRC) concur with the DOE's remedial action plan and certify that the remedial action complies with the standards promulgated by the US Environmental Protection Agency (EPA). The remedial action plan (RAP), which includes this remedial action selection report (RAS), has been developed to serve a two-fold purpose. First, it describes the activities that are proposed by the DOE to accomplish long-term stabilization and control of residual radioactive materials at the inactive uranium processing site near Lowman, Idaho. Second, this document and the remainder of the RAP, upon concurrence and execution by the DOE, the State of Idaho, and the NRC, becomes Appendix B of the Cooperative Agreement (No. DE-FC04-85AL20535) between the DOE and the State of Idaho.

  11. Remedial action plan and site design for stabilization of the inactive uranium mill tailings site at Lowman, Idaho: Remedial action selection report for the Lowman UMTRA project site, Idaho. Final report

    SciTech Connect

    Matthews, M.L.; Nagel, J.

    1991-09-01

    The inactive uranium mill tailings site near Lowman, Idaho, was designated as one of 24 abandoned uranium tailings sites to be remediated by the US Department of Energy (DOE) under the Uranium Mill Tailings Radiation Control Act of 1978 (UMTRCA). The UMTRCA requires that the US Nuclear Regulatory Commission (NRC) concur with the DOE`s remedial action plan and certify that the remedial action complies with the standards promulgated by the US Environmental Protection Agency (EPA). The remedial action plan (RAP), which includes this remedial action selection report (RAS), has been developed to serve a two-fold purpose. First, it describes the activities that are proposed by the DOE to accomplish long-term stabilization and control of residual radioactive materials at the inactive uranium processing site near Lowman, Idaho. Second, this document and the remainder of the RAP, upon concurrence and execution by the DOE, the State of Idaho, and the NRC, becomes Appendix B of the Cooperative Agreement (No. DE-FC04-85AL20535) between the DOE and the State of Idaho.

  12. A Pilot Study of the Interface Design of Cross-Cultural Web Sites through Usability Testing of Multilanguage Web Sites and Determining the Preferences of Taiwanese and American Users

    ERIC Educational Resources Information Center

    Ku, David Tawei; Chang, Chia-Chi

    2014-01-01

    By conducting usability testing on a multilanguage Web site, this study analyzed the cultural differences between Taiwanese and American users in the performance of assigned tasks. To provide feasible insight into cross-cultural Web site design, Microsoft Office Online (MOO) that supports both traditional Chinese and English and contains an almost…

  13. A Pilot Study of the Interface Design of Cross-Cultural Web Sites through Usability Testing of Multilanguage Web Sites and Determining the Preferences of Taiwanese and American Users

    ERIC Educational Resources Information Center

    Ku, David Tawei; Chang, Chia-Chi

    2014-01-01

    By conducting usability testing on a multilanguage Web site, this study analyzed the cultural differences between Taiwanese and American users in the performance of assigned tasks. To provide feasible insight into cross-cultural Web site design, Microsoft Office Online (MOO) that supports both traditional Chinese and English and contains an almost…

  14. Site-directed mutagenesis of the T4 endonuclease V gene: role of tyrosine-129 and -131 in pyrimidine dimer-specific binding

    SciTech Connect

    Stump, D.G.; Lloyd, R.S.

    1988-03-22

    T4 endonuclease V incises DNA at the sites of pyrimidine dimers through a two-step mechanism. These breakage reactions are preceded by the scanning of nontarget DNA and binding to pyrimidine dimers. In analogy to the synthetic tripeptides Lys-Trp-Lys and Lys-Tyr-Lys, which have been shown to be capable of producing single-strand scissions in DNA containing apurinic sites, endonuclease V has the amino acid sequence Trp-Tyr-Lys-Tyr-Tyr (128-132). Site-directed mutagenesis of the endonuclease V gene, denV, was performed at the Tyr-129 and at the Tyr-129 and Tyr-131 positions in order to convert the Tyr residues to nonaromatic amino acids to test their role in dimer-specific binding. The UV survival of repair-deficient (uvrA recA) Escherichia coli cells harboring the denV N-129 construction was dramatically reduced relative to wild-type denV+ cells. The survival of denV N-129,131 cells was indistinguishable from that of the parental strain lacking the denV gene. The mutant endonuclease V proteins were then characterized with regard to (1) dimer-specific nicking activity, (2) apurinic nicking activity, and (3) binding affinity to UV-irradiated DNA. Dimer-specific nicking activity and dimer-specific binding for both denV N-129 and N-129,131 were abolished, while apurinic-specific nicking was substantially retained in denV N-129,131 but was abolished in denV N-129. These results indicate that Tyr-129 and Tyr-131 positions of endonuclease V are at least important in pyrimidine dimer-specific binding and possibly nicking activity.

  15. Remedial action plan and site design for stabilization of the Inactive Uranium Mill Tailings Site, Maybell, Colorado. Final report, Appendixes to attachment 3

    SciTech Connect

    Not Available

    1994-06-01

    This document contains supporting appendices to attachment 3 for the remedial action and site stabilization plan for Maybell, Colorado UMTRA site. Appendix A includes the Hydrological Services Calculations and Appendix B contains Ground Water Quality by Location data.

  16. A designed DNA binding motif that recognizes extended sites and spans two adjacent major grooves†

    PubMed Central

    Rodríguez, Jéssica; Mosquera, Jesús; García-Fandiño, Rebeca; Vázquez, M. Eugenio; Mascareñas, José L.

    2016-01-01

    We report the rational design of a DNA-binding peptide construct composed of the DNA-contacting regions of two transcription factors (GCN4 and GAGA) linked through an AT-hook DNA anchor. The resulting chimera, which represents a new, non-natural DNA binding motif, binds with high affinity and selectivity to a long composite sequence of 13 base pairs (TCAT-AATT-GAGAG). PMID:27252825

  17. Active site similarity between human and Plasmodium falciparum phosphodiesterases: considerations for antimalarial drug design

    NASA Astrophysics Data System (ADS)

    Howard, Brittany L.; Thompson, Philip E.; Manallack, David T.

    2011-08-01

    The similarity between Plasmodium falciparum phosphodiesterase enzymes ( PfPDEs) and their human counterparts have been examined and human PDE9A was found to be a suitable template for the construction of homology models for each of the four PfPDE isoforms. In contrast, the architecture of the active sites of each model was most similar to human PDE1. Molecular docking was able to model cyclic guanosine monophosphate (cGMP) substrate binding in each case but a docking mode supporting cyclic adenosine monophosphate (cAMP) binding could not be found. Anticipating the potential of PfPDE inhibitors as anti-malarial drugs, a range of reported PDE inhibitors including zaprinast and sildenafil were docked into the model of PfPDEα. The results were consistent with their reported biological activities, and the potential of PDE1/9 inhibitor analogues was also supported by docking.

  18. Environmental monitoring at designed geopressured-geothermal well sites, Louisiana and Texas

    SciTech Connect

    Groat, C.; Stevenson, D.

    1990-01-01

    The research objectives of this report are to: implement and maintain the ongoing environmental monitoring program around DOE geopressured-geothermal test well in Louisiana and Texas; analyze and interpret collected data for evidence of subsidence and induced microearthquakes which may be brought about by geopressured-geothermal well testing and development; continue geological-geophysical studies of the Hulin and Gladys McCall sites incorporating new seismic data; continue review of previously identified and tested geopressured-geothermal prospects in Louisiana to determine if any link exists between such reservoirs and the existence of free gas in commercial or subcommercial quantities; and initiate review of geology, co-location and properties of geopressured brines with medium and heavy oil reservoirs in Louisiana utilizing existing maps, databases, reports, and journal articles.

  19. Environmental monitoring at designed geopressured-geothermal well sites, Louisiana and Texas

    SciTech Connect

    Not Available

    1991-01-01

    The research objectives of this report are to: implement and maintain the ongoing environmental monitoring program around DOE geopressured-geothermal test wells in Louisiana and Texas; analyze and interpret collected data for evidence of subsidence and induced microearthquakes which may be brought about by geopressured-geothermal well testing and development; continue geological-geophysical studies of the Hulin and Gladys McCall sites incorporating new seismic data; continue review of previously identified and tested geopressured-geothermal prospects in Louisiana to determine if any link exists between such reservoirs and the existence of free gas in commercial or subcommercial quantities; and initiate review of geology, co-location and properties of geopressured brines with medium and heavy oil reservoirs in Louisiana utilizing existing maps, databases, reports, and journal articles. 2 figs.

  20. Design, technology, and science: Sites for learning, resistance, and social reproduction in urban schools

    NASA Astrophysics Data System (ADS)

    Seiler, Gale; Tobin, Kenneth; Sokolic, Joseph

    2001-09-01

    The teaching of science through activities that emphasize design and technology has been advocated as a vehicle for accomplishing science for all students. This study was situated in an inner7-city neighborhood school populated mainly by African American students from life worlds characterized by poverty. The article explores the discourse and practices of students and three coteachers as a curriculum was enacted to provide opportunities for students to learn about the physics of motion through designing, building, and testing a model car. Some students participated in ways that led to their building viable model cars and interacting with one another in ways that suggest design and technological competence. However, there also was evidence of resistance from students who participated sporadically and refused to cooperate with teachers as they endeavored to structure the environment in ways that would lead to a deeper understanding of science. Analysis of in-class interactions reveals an untapped potential for the emergence of a sciencelike discourse and diverse outcomes. Among the challenges explored in this article is a struggle for respect that permeates the students' lives on the street and bleeds into the classroom environment. Whereas teachers enacted the curriculum as if learning was the chief goal for students, it is apparent that students used the class opportunistically to maintain and earn the respect of peers.

  1. Eggs in the Freezer: Energetic Consequences of Nest Site and Nest Design in Arctic Breeding Shorebirds

    PubMed Central

    Tulp, Ingrid; Schekkerman, Hans; de Leeuw, Joep

    2012-01-01

    Birds construct nests for several reasons. For species that breed in the Arctic, the insulative properties of nests are very important. Incubation is costly there and due to an increasing surface to volume ratio, more so in smaller species. Small species are therefore more likely to place their nests in thermally favourable microhabitats and/or to invest more in nest insulation than large species. To test this hypothesis, we examined characteristics of nests of six Arctic breeding shorebird species. All species chose thermally favourable nesting sites in a higher proportion than expected on the basis of habitat availability. Site choice did not differ between species. Depth to frozen ground, measured near the nests, decreased in the course of the season at similar non-species-specific speeds, but this depth increased with species size. Nest cup depth and nest scrape depth (nest cup without the lining) were unrelated to body mass (we applied an exponent of 0.73, to account for metabolic activity of the differently sized species). Cup depth divided by diameter2 was used as a measure of nest cup shape. Small species had narrow and deep nests, while large species had wide shallow nests. The thickness of nest lining varied between 0.1 cm and 7.6 cm, and decreased significantly with body mass. We reconstruct the combined effect of different nest properties on the egg cooling coefficient using previously published quantitative relationships. The predicted effect of nest cup depth and lining depth on heat loss to the frozen ground did not correlate with body mass, but the sheltering effect of nest cup diameter against wind and the effects of lining material on the cooling coefficient increased with body mass. Our results suggest that small arctic shorebirds invest more in the insulation of their nests than large species. PMID:22701596

  2. In silico designing of a new cysteine analogue of hirudin variant 3 for site specific PEGylation

    PubMed Central

    Sajjadi, Seyed Mehdi; Rahimi, Hamzeh; Mohammadi, Saeed; Faranoush, Mohammad; Mirzahoseini, Hasan; Toogeh, Gholamreza

    2017-01-01

    Hirudin is an anticoagulant agent of the salivary glands of the medicinal leech. Recombinant hirudin (r-Hir) displays certain drawbacks including bleeding and immunogenicity. To solve these problems, cysteine-specific PEGylation has been proposed as a successful technique. However, proper selection of the appropriate cysteine residue for substitution is a critical step. This study has, for the first time, used a computational approach aimed at identifying a single potential PEGylation site for replacement by cysteine residue in the hirudin variant 3 (HV3). Homology modeling (HM) was performed using MODELLER. All non-cysteine residues of the HV3 were replaced with the cysteine. The best model was selected based on the results of discrete optimized protein energy score, PROCHECK software, and Verify3D. The receptor binding was investigated using protein-protein docking by ClusPro web tool which was then visualized using LigPlot+ software and PyMOL. Finally, multiple sequence alignment (MSA) using ClustalW software and disulfide bond prediction were performed. According to the results of HM and docking, Q33C, which was located on the surface of the protein, was the best site for PEGylation. Furthermore, MSA showed that Q33 was not a conserved residue and LigPlot+ software showed that it is not involved in the hirudin-thrombin binding pocket. Moreover, prediction softwares established that it is not involved in disulfide bond formation. In this study, for the first time, the utility of the in silico approach for creating a cysteine analogue of HV3 was introduced. Our study demonstrated that the substitution of Q33 by cysteine probably has no effect on the biological activity of the HV3. However, experimental analyses are required to confirm the results. PMID:28255315

  3. Elucidating the design principles of photosynthetic electron-transfer proteins by site-directed spin labeling EPR spectroscopy.

    PubMed

    Ishara Silva, K; Jagannathan, Bharat; Golbeck, John H; Lakshmi, K V

    2016-05-01

    Site-directed spin labeling electron paramagnetic resonance (SDSL EPR) spectroscopy is a powerful tool to determine solvent accessibility, side-chain dynamics, and inter-spin distances at specific sites in biological macromolecules. This information provides important insights into the structure and dynamics of both natural and designed proteins and protein complexes. Here, we discuss the application of SDSL EPR spectroscopy in probing the charge-transfer cofactors in photosynthetic reaction centers (RC) such as photosystem I (PSI) and the bacterial reaction center (bRC). Photosynthetic RCs are large multi-subunit proteins (molecular weight≥300 kDa) that perform light-driven charge transfer reactions in photosynthesis. These reactions are carried out by cofactors that are paramagnetic in one of their oxidation states. This renders the RCs unsuitable for conventional nuclear magnetic resonance spectroscopy investigations. However, the presence of native paramagnetic centers and the ability to covalently attach site-directed spin labels in RCs makes them ideally suited for the application of SDSL EPR spectroscopy. The paramagnetic centers serve as probes of conformational changes, dynamics of subunit assembly, and the relative motion of cofactors and peptide subunits. In this review, we describe novel applications of SDSL EPR spectroscopy for elucidating the effects of local structure and dynamics on the electron-transfer cofactors of photosynthetic RCs. Because SDSL EPR Spectroscopy is uniquely suited to provide dynamic information on protein motion, it is a particularly useful method in the engineering and analysis of designed electron transfer proteins and protein networks. This article is part of a Special Issue entitled Biodesign for Bioenergetics--the design and engineering of electronic transfer cofactors, proteins and protein networks, edited by Ronald L. Koder and J.L. Ross Anderson.

  4. Rational design of carbon nitride photocatalysts by identification of cyanamide defects as catalytically relevant sites

    PubMed Central

    Lau, Vincent Wing-hei; Moudrakovski, Igor; Botari, Tiago; Weinberger, Simon; Mesch, Maria B.; Duppel, Viola; Senker, Jürgen; Blum, Volker; Lotsch, Bettina V.

    2016-01-01

    The heptazine-based polymer melon (also known as graphitic carbon nitride, g-C3N4) is a promising photocatalyst for hydrogen evolution. Nonetheless, attempts to improve its inherently low activity are rarely based on rational approaches because of a lack of fundamental understanding of its mechanistic operation. Here we employ molecular heptazine-based model catalysts to identify the cyanamide moiety as a photocatalytically relevant ‘defect'. We exploit this knowledge for the rational design of a carbon nitride polymer populated with cyanamide groups, yielding a material with 12 and 16 times the hydrogen evolution rate and apparent quantum efficiency (400 nm), respectively, compared with the unmodified melon. Computational modelling and material characterization suggest that this moiety improves coordination (and, in turn, charge transfer kinetics) to the platinum co-catalyst and enhances the separation of the photogenerated charge carriers. The demonstrated knowledge transfer for rational catalyst design presented here provides the conceptual framework for engineering high-performance heptazine-based photocatalysts. PMID:27387536

  5. Rational design of carbon nitride photocatalysts by identification of cyanamide defects as catalytically relevant sites

    NASA Astrophysics Data System (ADS)

    Lau, Vincent Wing-Hei; Moudrakovski, Igor; Botari, Tiago; Weinberger, Simon; Mesch, Maria B.; Duppel, Viola; Senker, Jürgen; Blum, Volker; Lotsch, Bettina V.

    2016-07-01

    The heptazine-based polymer melon (also known as graphitic carbon nitride, g-C3N4) is a promising photocatalyst for hydrogen evolution. Nonetheless, attempts to improve its inherently low activity are rarely based on rational approaches because of a lack of fundamental understanding of its mechanistic operation. Here we employ molecular heptazine-based model catalysts to identify the cyanamide moiety as a photocatalytically relevant `defect'. We exploit this knowledge for the rational design of a carbon nitride polymer populated with cyanamide groups, yielding a material with 12 and 16 times the hydrogen evolution rate and apparent quantum efficiency (400 nm), respectively, compared with the unmodified melon. Computational modelling and material characterization suggest that this moiety improves coordination (and, in turn, charge transfer kinetics) to the platinum co-catalyst and enhances the separation of the photogenerated charge carriers. The demonstrated knowledge transfer for rational catalyst design presented here provides the conceptual framework for engineering high-performance heptazine-based photocatalysts.

  6. CRISPRdirect: software for designing CRISPR/Cas guide RNA with reduced off-target sites

    PubMed Central

    Naito, Yuki; Hino, Kimihiro; Bono, Hidemasa; Ui-Tei, Kumiko

    2015-01-01

    Summary: CRISPRdirect is a simple and functional web server for selecting rational CRISPR/Cas targets from an input sequence. The CRISPR/Cas system is a promising technique for genome engineering which allows target-specific cleavage of genomic DNA guided by Cas9 nuclease in complex with a guide RNA (gRNA), that complementarily binds to a ∼20 nt targeted sequence. The target sequence requirements are twofold. First, the 5′-NGG protospacer adjacent motif (PAM) sequence must be located adjacent to the target sequence. Second, the target sequence should be specific within the entire genome in order to avoid off-target editing. CRISPRdirect enables users to easily select rational target sequences with minimized off-target sites by performing exhaustive searches against genomic sequences. The server currently incorporates the genomic sequences of human, mouse, rat, marmoset, pig, chicken, frog, zebrafish, Ciona, fruit fly, silkworm, Caenorhabditis elegans, Arabidopsis, rice, Sorghum and budding yeast. Availability: Freely available at http://crispr.dbcls.jp/. Contact: y-naito@dbcls.rois.ac.jp Supplementary information: Supplementary data are available at Bioinformatics online. PMID:25414360

  7. Does wine glass size influence sales for on-site consumption? A multiple treatment reversal design.

    PubMed

    Pechey, Rachel; Couturier, Dominique-Laurent; Hollands, Gareth J; Mantzari, Eleni; Munafò, Marcus R; Marteau, Theresa M

    2016-06-07

    Wine glass size can influence both perceptions of portion size and the amount poured, but its impact upon purchasing and consumption is unknown. This study aimed to examine the impact of wine glass size on wine sales for on-site consumption, keeping portion size constant. In one establishment (with separate bar and restaurant areas) in Cambridge, England, wine glass size (Standard; Larger; Smaller) was changed over eight fortnightly periods. The bar and restaurant differ in wine sales by the glass vs. by the bottle (93 % vs. 63 % by the glass respectively). Daily wine volume purchased was 9.4 % (95 % CI: 1.9, 17.5) higher when sold in larger compared to standard-sized glasses. This effect seemed principally driven by sales in the bar area (bar: 14.4 % [3.3, 26.7]; restaurant: 8.2 % [-2.5, 20.1]). Findings were inconclusive as to whether sales were different with smaller vs. standard-sized glasses. The size of glasses in which wine is sold, keeping the portion size constant, can affect consumption, with larger glasses increasing consumption. The hypothesised mechanisms for these differential effects need to be tested in a replication study. If replicated, policy implications could include considering glass size amongst alcohol licensing requirements. ISRCTN registry: ISRCTN12018175 . Registered 12(th) May 2015.

  8. Design of a soil cutting resistance sensor for application in site-specific tillage.

    PubMed

    Agüera, Juan; Carballido, Jacob; Gil, Jesús; Gliever, Chris J; Perez-Ruiz, Manuel

    2013-05-10

    One objective of precision agriculture is to provide accurate information about soil and crop properties to optimize the management of agricultural inputs to meet site-specific needs. This paper describes the development of a sensor equipped with RTK-GPS technology that continuously and efficiently measures soil cutting resistance at various depths while traversing the field. Laboratory and preliminary field tests verified the accuracy of this prototype soil strength sensor. The data obtained using a hand-operated soil cone penetrometer was used to evaluate this field soil compaction depth profile sensor. To date, this sensor has only been tested in one field under one gravimetric water content condition. This field test revealed that the relationships between the soil strength profile sensor (SSPS) cutting force and soil cone index values are assumed to be quadratic for the various depths considered: 0-10, 10-20 and 20-30 cm (r2 = 0.58, 0.45 and 0.54, respectively). Soil resistance contour maps illustrated its practical value. The developed sensor provides accurate, timely and affordable information on soil properties to optimize resources and improve agricultural economy.

  9. Design of a Soil Cutting Resistance Sensor for Application in Site-Specific Tillage

    PubMed Central

    Agüera, Juan; Carballido, Jacob; Gil, Jesús; Gliever, Chris J.; Perez-Ruiz, Manuel

    2013-01-01

    One objective of precision agriculture is to provide accurate information about soil and crop properties to optimize the management of agricultural inputs to meet site-specific needs. This paper describes the development of a sensor equipped with RTK-GPS technology that continuously and efficiently measures soil cutting resistance at various depths while traversing the field. Laboratory and preliminary field tests verified the accuracy of this prototype soil strength sensor. The data obtained using a hand-operated soil cone penetrometer was used to evaluate this field soil compaction depth profile sensor. To date, this sensor has only been tested in one field under one gravimetric water content condition. This field test revealed that the relationships between the soil strength profile sensor (SSPS) cutting force and soil cone index values are assumed to be quadratic for the various depths considered: 0–10, 10–20 and 20–30 cm (r2 = 0.58, 0.45 and 0.54, respectively). Soil resistance contour maps illustrated its practical value. The developed sensor provides accurate, timely and affordable information on soil properties to optimize resources and improve agricultural economy. PMID:23666127

  10. Design of the primary pre-TRMM and TRMM ground truth site

    NASA Technical Reports Server (NTRS)

    Garstang, Michael

    1988-01-01

    The primary objective of the Tropical Rain Measuring Mission (TRMM) were to: integrate the rain gage measurements with radar measurements of rainfall using the KSFC/Patrick digitized radar and associated rainfall network; delineate the major rain bearing systems over Florida using the Weather Service reported radar/rainfall distributions; combine the integrated measurements with the delineated rain bearing systems; use the results of the combined measurements and delineated rain bearing systems to represent patterns of rainfall which actually exist and contribute significantly to the rainfall to test sampling strategies and based on the results of these analyses decide upon the ground truth network; and complete the design begun in Phase 1 of a multi-scale (space and time) surface observing precipitation network centered upon KSFC. Work accomplished and in progress is discussed.

  11. Design of accurate predictors for DNA-binding sites in proteins using hybrid SVM-PSSM method.

    PubMed

    Ho, Shinn-Ying; Yu, Fu-Chieh; Chang, Chia-Yun; Huang, Hui-Ling

    2007-01-01

    In this paper, we investigate the design of accurate predictors for DNA-binding sites in proteins from amino acid sequences. As a result, we propose a hybrid method using support vector machine (SVM) in conjunction with evolutionary information of amino acid sequences in terms of their position-specific scoring matrices (PSSMs) for prediction of DNA-binding sites. Considering the numbers of binding and non-binding residues in proteins are significantly unequal, two additional weights as well as SVM parameters are analyzed and adopted to maximize net prediction (NP, an average of sensitivity and specificity) accuracy. To evaluate the generalization ability of the proposed method SVM-PSSM, a DNA-binding dataset PDC-59 consisting of 59 protein chains with low sequence identity on each other is additionally established. The SVM-based method using the same six-fold cross-validation procedure and PSSM features has NP=80.15% for the training dataset PDNA-62 and NP=69.54% for the test dataset PDC-59, which are much better than the existing neural network-based method by increasing the NP values for training and test accuracies up to 13.45% and 16.53%, respectively. Simulation results reveal that SVM-PSSM performs well in predicting DNA-binding sites of novel proteins from amino acid sequences.

  12. MDC-Analyzer: a novel degenerate primer design tool for the construction of intelligent mutagenesis libraries with contiguous sites.

    PubMed

    Tang, Lixia; Wang, Xiong; Ru, Beibei; Sun, Hengfei; Huang, Jian; Gao, Hui

    2014-06-01

    Recent computational and bioinformatics advances have enabled the efficient creation of novel biocatalysts by reducing amino acid variability at hot spot regions. To further expand the utility of this strategy, we present here a tool called Multi-site Degenerate Codon Analyzer (MDC-Analyzer) for the automated design of intelligent mutagenesis libraries that can completely cover user-defined randomized sequences, especially when multiple contiguous and/or adjacent sites are targeted. By initially defining an objective function, the possible optimal degenerate PCR primer profiles could be automatically explored using the heuristic approach of Greedy Best-First-Search. Compared to the previously developed DC-Analyzer, MDC-Analyzer allows for the existence of a small amount of undesired sequences as a tradeoff between the number of degenerate primers and the encoded library size while still providing all the benefits of DC-Analyzer with the ability to randomize multiple contiguous sites. MDC-Analyzer was validated using a series of randomly generated mutation schemes and experimental case studies on the evolution of halohydrin dehalogenase, which proved that the MDC methodology is more efficient than other methods and is particularly well-suited to exploring the sequence space of proteins using data-driven protein engineering strategies.

  13. A versatile photoactivatable probe designed to label the diphosphate binding site of farnesyl diphosphate utilizing enzymes.

    PubMed

    Henry, Olivier; Lopez-Gallego, Fernando; Agger, Sean A; Schmidt-Dannert, Claudia; Sen, Stephanie; Shintani, David; Cornish, Katrina; Distefano, Mark D

    2009-07-01

    Farnesyl diphosphate (FPP) is a substrate for a diverse number of enzymes found in nature. Photoactive analogues of isoprenoid diphosphates containing either benzophenone, diazotrifluoropropionate or azide groups have been useful for studying both the enzymes that synthesize FPP as well as those that employ FPP as a substrate. Here we describe the synthesis and properties of a new class of FPP analogues that links an unmodified farnesyl group to a diphosphate mimic containing a photoactive benzophenone moiety; thus, importantly, these compounds are photoactive FPP analogues that contain no modifications of the isoprenoid portion of the molecule that may interfere with substrate binding in the active site of an FPP utilizing enzyme. Two isomeric compounds containing meta- and para-substituted benzophenones were prepared. These two analogues inhibit Saccharomyces cerevisiae protein farnesyltransferase (ScPFTase) with IC(50) values of 5.8 (meta isomer) and 3.0 microM (para isomer); the more potent analogue, the para isomer, was shown to be a competitive inhibitor of ScPFTase with respect to FPP with a K(I) of 0.46 microM. Radiolabeled forms of both analogues selectively labeled the beta-subunit of ScPFTase. The para isomer was also shown to label Escherichia coli farnesyl diphosphate synthase and Drosophila melanogaster farnesyl diphosphate synthase. Finally, the para isomer was shown to be an alternative substrate for a sesquiterpene synthase from Nostoc sp. strain PCC7120, a cyanobacterial source; the compound also labeled the purified enzyme upon photolysis. Taken together, these results using a number of enzymes demonstrate that this new class of probes should be useful for a plethora of studies of FPP-utilizing enzymes.

  14. Use of Ground Penetrating Radar for Site Investigation of Low-Volume Roadways and Design Recommendations

    NASA Astrophysics Data System (ADS)

    Scullion, T.; Saarenketo, T.

    2002-07-01

    This report will present several case studies describing the use of ground penetrating radar (GPR) technology for site investigations. Two types of GPR will be described-the air-launched and ground-coupled systems. The use of air-launched radar is well established within the Texas Department of Transportation (TxDOT). The limitation of this technology is its depth of penetration. While providing very useful information on the surface and base layers, it provides little information on the sub-grade soils. The use of low-frequency ground-coupled radar systems will provide little useful near-surface information but it can provide data on sub-grade properties and how they vary along a project. Combining both radar types can potentially provide a comprehensive subsurface investigative tool for both new pavement construction and for major pavement rehabilitation projects. In this report a brief description will be provided of the different systems together with the software used to process the GPR signals. Air-launched data are processed with the COLORMAP system developed by the Texas Transportation Institute. The ground-coupled data are processed using the Road Doctor system developed by Roadscanners, Inc. of Finland. The case studies presented were collected on actual TxDOT evaluation projects mainly in the Bryan District. They range from near-surface applications where the goal was to identify changes in pavement structure which were not available in construction records to identifying the areas beneath the pavement subsidence associated with strip mining activities.

  15. Energy conserving site design: Greenbrier case study, Chesapeake, Virginia. Final report

    SciTech Connect

    Not Available

    1980-04-01

    A specific case study of project planning for energy conservation for a major planned unit development at the 3000-acre Greenbrier development site in Chesapeake, Virginia, is summarized. The research suggests that very considerable reductions in energy conservation can be achieved within the confines of private-sector land development and residential construction with increased incremental costs of $200.00 to $3150.00 per dwelling unit. It is hypothesized that energy consumption at Greenbrier can be reduced by one-half with an average annual savings of 21,275 kWh per residential unit, using state-of-the-art technology with careful planning and control. This represents an annual savings $750.00 per unit at the current utility rate of 3.5 cents per kWh. These savings can be achieved through reduction in heating and cooling loads and application of more-efficient heating and cooling of the remaining loads. The reduction in loads are achieved by redesign of the land plan to include a higher percentage of south-facing lots, use of vegetation to modify microclimate, decreases in air infiltration, the use of 2 x 6 framing, better insulation, and the use of an insulated slab-on-grade foundation. Further energy savings can be expected by increased efficiencies in mechanical systems used for space heating and cooling and domestic hot water. When applied to the single-family portion of Greenbrier, containing 541 dwelling units, these options reduce the total end-use energy consumption 54.7%. This reduction represents an annual savings of $432,800.00 for an initial capital investment of $1.7 million.

  16. A Versatile Photoactivatable Probe Designed to Label the Diphosphate Binding Site of Farnesyl Diphosphate Utilizing Enzymes

    PubMed Central

    Henry, Olivier; Lopez-Gallego, Fernando; Agger, Sean A.; Schmidt-Dannert, Claudia; Sen, Stephanie; Shintani, David; Cornish, Katrina; Distefano, Mark D.

    2009-01-01

    Farnesyl diphosphate (FPP) is a substrate for a diverse number of enzymes found in nature. Photoactive analogues of isoprenoid diphosphates containing either benzophenone, diazotrifluropropionate or azide groups have been useful for studying both the enzymes that synthesize FPP as well as those that employ FPP as a substrate. Here we describe the synthesis and properties of a new class of FPP analogues that links an unmodified farnesyl group to a diphosphate mimic containing a photoactive benzophenone moiety; thus, importantly, these compounds are photoactive FPP analogues that contain no modifications of the isoprenoid portion of the molecule that may interfere with substrate binding in the active site of an FPP utilizing enzyme. Two isomeric compounds containing meta- and para-substituted benzophenones were prepared. These two analogues inhibit S. cerevisiae protein farnesyltransferase (ScPFTase) with IC50 values of 5.8 (meta isomer) and 3.0 µM (para isomer); the more potent analogue, the para isomer, was shown to be a competitive inhibitor of ScPFTase with respect to FPP with a KI of 0.46 µM. Radiolabeled forms of both analogues selectively labelled the β-subunit of ScPFTase. The para isomer was also shown to label E. coli farnesyl diphosphate synthase and Drosophila melanogaster farnesyl diphosphate synthase. Finally, the para isomer was shown to be an alternative substrate for a sesquiterpene synthase from Nostoc sp. strain PCC7120, a cyanobacterial source; the compound also labeled the purified enzyme upon photolysis. Taken together, these results using a number of enzymes demonstrate that this new class of probes should be useful for a plethora of studies of FPP-utilizing enzymes. PMID:19447628

  17. "Intelligent design" of a 3D reflection survey for the SAFOD drill-hole site

    NASA Astrophysics Data System (ADS)

    Alvarez, G.; Hole, J. A.; Klemperer, S. L.; Biondi, B.; Imhof, M.

    2003-12-01

    SAFOD seeks to better understand the earthquake process by drilling though the San Andreas fault (SAF) to sample an earthquake in situ. To capitalize fully on the opportunities presented by the 1D drill-hole into a complex fault zone we must characterize the surrounding 3D geology at a scale commensurate with the drilling observations, to provide the structural context to extrapolate 1D drilling results along the fault plane and into the surrounding 3D volume. Excellent active-2D and passive-3D seismic observations completed and underway lack the detailed 3D resolution required. Only an industry-quality 3D reflection survey can provide c. 25 m subsurface sample-spacing horizontally and vertically. A 3D reflection survey will provide subsurface structural and stratigraphic control at the 100-m level, mapping major geologic units, structural boundaries, and subsurface relationships between the many faults that make up the SAF fault system. A principal objective should be a reflection-image (horizon-slice through the 3D volume) of the near-vertical fault plane(s) to show variations in physical properties around the drill-hole. Without a 3D reflection image of the fault zone, we risk interpreting drilled anomalies as ubiquitous properties of the fault, or risk missing important anomalies altogether. Such a survey cannot be properly costed or technically designed without major planning. "Intelligent survey design" can minimize source and receiver effort without compromising data-quality at the fault target. Such optimization can in principal reduce the cost of a 3D seismic survey by a factor of two or three, utilizing the known surface logistic constraints, partially-known sub-surface velocity field, and the suite of scientific targets at SAFOD. Our methodology poses the selection of the survey parameters as an optimization process that allows the parameters to vary spatially in response to changes in the subsurface. The acquisition geometry is locally optimized for

  18. Design and Analysis of Map Relative Localization for Access to Hazardous Landing Sites on Mars

    NASA Technical Reports Server (NTRS)

    Johnson, Andrew E.; Aaron, Seth; Cheng, Yang; Montgomery, James; Trawny, Nikolas; Tweddle, Brent; Vaughan, Geoffrey; Zheng, Jason

    2016-01-01

    Human and robotic planetary lander missions require accurate surface relative position knowledge to land near science targets or next to pre-deployed assets. In the absence of GPS, accurate position estimates can be obtained by automatically matching sensor data collected during descent to an on-board map. The Lander Vision System (LVS) that is being developed for Mars landing applications generates landmark matches in descent imagery and combines these with inertial data to estimate vehicle position, velocity and attitude. This paper describes recent LVS design work focused on making the map relative localization algorithms robust to challenging environmental conditions like bland terrain, appearance differences between the map and image and initial input state errors. Improved results are shown using data from a recent LVS field test campaign. This paper also fills a gap in analysis to date by assessing the performance of the LVS with data sets containing significant vertical motion including a complete data set from the Mars Science Laboratory mission, a Mars landing simulation, and field test data taken over multiple altitudes above the same scene. Accurate and robust performance is achieved for all data sets indicating that vertical motion does not play a significant role in position estimation performance.

  19. Designating Earth's Moon as a United Nations World Heritage Site - Permanently Protected from Commercial or Military Uses

    NASA Astrophysics Data System (ADS)

    Steiner, R. G.

    2002-01-01

    This paper proposes that Earth's Moon, in its entirety, be designated a United Nations World Heritage Site (WHS), permanently protected from any and all commercial or military utilization and reserved exclusively for scientific and aesthetic purposes. The paper discusses: 1) the extraordinary importance of the Moon for science, culture, and religion - past, present and future; 2) the history of proposals to exploit the Moon for commercial and military purposes and the shortcomings of this colonial, exploitation paradigm; and 3) the necessity, policy mechanisms, and political dynamics of designating the Moon as a World Heritage Site, permanently protected from commercial and/or military uses. The first part of the paper discusses the extraordinary importance of the Moon as it exists today - as a scientific laboratory, a source of beauty and inspiration throughout human evolution, a source for artistic expression, and as an object that is considered sacred by many cultures. Next, the paper traces the history of specific proposals for the exploitation of the Moon for commercial and/or military purposes - including plans by the U.S. Air Force in 1959 to detonate a nuclear explosion on the Moon, proposals to strip-mine the lunar regolith for helium-3 and rocket-fuel hydrogen; construction of solar power plants to transmit energy to Earth, and proposals to use the lunar surface as a billboard upon which to project commercial advertisements visible from Earth. The profound ethical, legal, and scientific shortcomings of this exploitation paradigm are described as an emerging Extraterrestrial Manifest Destiny that we have a collective obligation to challenge and constrain. The paper proposes that space exploration be infused with an ethical commitment to compassion, reverence, conservation, and non-interference to abiotic and biotic systems alike; as opposed to the expansion and extraterrestrial imposition of the colonization, exploitation, domination, and despoliation

  20. Compressed air energy storage: Preliminary design and site development program in an aquifer. Task 1: Establish facility design criteria and utility benefits

    NASA Astrophysics Data System (ADS)

    1980-10-01

    Compressed air energy storage (CAES) stores mechanical energy in the form of compressed air during off-peak hours, using power supplied by a large, high efficiency baseload power plant. At times of high electrical demand, the compressed air is drawn from storage and is heated in a combustor by the burning of fuel oil, after which the air is expanded in a turbine. Essentially all of the turbine output can be applied to the generation of electricity, unlike a conventional gas turbine which expends approximately two-thirds of the turbine shaft power in driving the air compressor. The separation of the compression and generation modes in the system results in increased net generation and greater premium fuel economy. Work performed in establishing facility design criteria for a CAES system with aquifer storage includes: determination of initial design bases; preliminary analysis of the CAES system; development of data for site-specific analysis of the CAES system; detailed analysis of the CAES system for three selected heat cycles; CAES power plant design; and an economic analysis of CAES.

  1. Compressed air energy storage: preliminary design and site development program in an aquifer. Final draft, Task 1: establish facility design criteria and utility benefits

    SciTech Connect

    1980-10-01

    Compressed air energy storage (CAES) has been identified as one of the principal new energy storage technologies worthy of further research and development. The CAES system stores mechanical energy in the form of compressed air during off-peak hours, using power supplied by a large, high-efficiency baseload power plant. At times of high electrical demand, the compressed air is drawn from storage and is heated in a combustor by the burning of fuel oil, after which the air is expanded in a turbine. In this manner, essentially all of the turbine output can be applied to the generation of electricity, unlike a conventional gas turbine which expends approximately two-thirds of the turbine shaft power in driving the air compressor. The separation of the compression and generation modes in the CAES system results in increased net generation and greater premium fuel economy. The use of CAES systems to meet the utilities' high electrical demand requirements is particularly attractive in view of the reduced availability of premium fuels such as oil and natural gas. This volume documents the Task 1 work performed in establishing facility design criteria for a CAES system with aquifer storage. Information is included on: determination of initial design bases; preliminary analysis of the CAES system; development of data for site-specific analysis of the CAES system; detailed analysis of the CAES system for three selected heat cycles; CAES power plant design; and an economic analysis of CAES.

  2. Remedial action plan and site design for stabilization of the inactive uranium mill tailings sites at Slick Rock, Colorado. Attachment 3, Ground water hydrology report: Preliminary final

    SciTech Connect

    Not Available

    1994-03-04

    The US Environmental Protection Agency (EPA) has established health and environmental protection regulations to correct and prevent ground water contamination resulting from processing activities at inactive uranium milling sites (52 FR 36000 (1987)). According to the Uranium Mill Tailings Radiation Control Act (UMTRCA) of 1978, 42 USC {section}7901 et seq., the US Department of Energy (DOE) is responsible for assessing the inactive uranium processing sites. The DOE has determined that for Slick Rock, this assessment shall include hydrogeologic site characterization for two separate uranium processing sites, the Union Carbide (UC) site and the North Continent (NC) site, and for the proposed Burro Canyon disposal site. The water resources protection strategy that describes how the proposed action will comply with the EPA ground water protection standards is presented in Attachment 4. The following site characterization activities are discussed in this attachment: Characterization of the hydrogeologic environment, including hydrostratigraphy, ground water occurrence, aquifer parameters, and areas of recharge and discharge. Characterization of existing ground water quality by comparison with background water quality and the maximum concentration limits (MCL) of the proposed EPA ground water protection standards. Definition of physical and chemical characteristics of the potential contaminant source, including concentration and leachability of the source in relation to migration in ground water and hydraulically connected surface water. Description of local water resources, including current and future use, availability, and alternative supplies.

  3. Reports of Public Scoping Meetings for the Supplemental Environmental Impact Statement for the Designation of Dredged Material Disposal Sites in Eastern Long Island Sound

    EPA Pesticide Factsheets

    These reports provide summaries of the scoping meetings as part of the Supplemental Environmental Impact Statement (SEIS) process for the designation of dredged material disposal sites in Eastern Long Island Sound.

  4. Bioinspired Design and Oriented Synthesis of Immunogenic Site-Specifically Penicilloylated Peptides.

    PubMed

    Scornet, Noémie; Delarue-Cochin, Sandrine; Azoury, Marie Eliane; Le Mignon, Maxime; Chemelle, Julie-Anne; Nony, Emmanuel; Maillère, Bernard; Terreux, Raphaël; Pallardy, Marc; Joseph, Delphine

    2016-11-16

    β-Lactam antibiotics allergy is recognized as a public health concern. By covalently binding to serum proteins, penicillins are known to form immunogenic complexes. The latter are recognized and digested by antigen-presenting cells into drug-hapten peptides leading to the immunization of treated persons and IgE-mediated hypersensitivity reactions encompassing anaphylaxis. If type I allergic reactions to drugs are often unpredictable, they are known to be dependent on CD4(+) T-cells. This fundamental study revisits the chemical basis of the benzylpenicillin (BP) allergy with the aim of identifying immunologically relevant biomimetic benzylpenicilloylated peptides through the analysis of BP-conjugated human serum albumin (BP-HSA) profile and the evaluation of the naı̈ve CD4(+) T-cell responses to candidate BP-HSA-derived peptides. The chemical structures of BP-HSA bioconjugates synthesized in vitro at both physiological and basic pH were investigated by mass spectrometry. From the ten most representative lysine residues grafted by BP-hapten, HSA-bioinspired 15-mer peptide sequences were designed and the potential T-cell epitope profile of each peptide was predicted using two complementary in silico approaches, i.e., HLA class II binding prediction tools from the Immune Epitope Database and Analysis Resource (IEDB) and computational alanine scanning mutagenesis. Twelve structurally diversified benzylpenicilloylated peptides (BP-Ps) were selected and synthesized with the aid of a flexible synthesis pathway using an original benzylpenicilloylated lysine monomer as common precursor. In order to corroborate their predicted "epitope" profile, the naı̈ve CD4(+) T-cell response specific to BP was evaluated through a coculture approach. To our knowledge, this study showed for the first time the ability of bioinspired peptides structurally stemming from BP-HSA to be recognized by naı̈ve CD4(+) T-cells thus identifying a pre-existing T-cell repertoire for penicillin

  5. Fragment-Based Design of Ligands Targeting a Novel Site on the Integrase Enzyme of Human Immunodeficiency Virus;#8197;1

    SciTech Connect

    Wielens, Jerome; Headey, Stephen J.; Deadman, John J.; Rhodes, David I.; Parker, Michael W.; Chalmers, David K.; Scanlon, Martin J.

    2011-08-17

    Fragment-based screening has been used to identify a novel ligand binding site on HIV-1 integrase. Crystal structures of fragments bound at this site (shown) have been used to design elaborated second-generation compounds that bind with higher affinity and good ligand efficiency.

  6. Learning to Design and Implement Educational Web Sites within Pre-Service Training: a Project-Based Learning Environment and its Impact on Student Teachers

    ERIC Educational Resources Information Center

    Papastergiou, Marina

    2005-01-01

    This paper reports on a project-based learning (PBL) environment targeted towards introducing student teachers to the design and development of educational web sites. The participants were 46 student teachers who constructed 16 web sites for primary school. A survey of their views, before and after the projects, and an evaluation of the web sites…

  7. Learning to Design and Implement Educational Web Sites within Pre-Service Training: A Project-Based Learning Environment and Its Impact on Student Teachers

    ERIC Educational Resources Information Center

    Papastergiou, Marina

    2005-01-01

    This paper reports on a project-based learning (PBL) environment targeted towards introducing student teachers to the design and development of educational web sites. The participants were 46 student teachers who constructed 16 web sites for primary school. A survey of their views, before and after the projects, and an evaluation of the web sites…

  8. Learning to Design and Implement Educational Web Sites within Pre-Service Training: A Project-Based Learning Environment and Its Impact on Student Teachers

    ERIC Educational Resources Information Center

    Papastergiou, Marina

    2005-01-01

    This paper reports on a project-based learning (PBL) environment targeted towards introducing student teachers to the design and development of educational web sites. The participants were 46 student teachers who constructed 16 web sites for primary school. A survey of their views, before and after the projects, and an evaluation of the web sites…

  9. Learning to Design and Implement Educational Web Sites within Pre-Service Training: a Project-Based Learning Environment and its Impact on Student Teachers

    ERIC Educational Resources Information Center

    Papastergiou, Marina

    2005-01-01

    This paper reports on a project-based learning (PBL) environment targeted towards introducing student teachers to the design and development of educational web sites. The participants were 46 student teachers who constructed 16 web sites for primary school. A survey of their views, before and after the projects, and an evaluation of the web sites…

  10. Annual survival estimation of migratory songbirds confounded by incomplete breeding site-fidelity: Study designs that may help

    USGS Publications Warehouse

    Marshall, M.R.; Diefenbach, D.R.; Wood, L.A.; Cooper, R.J.

    2004-01-01

    Many species of bird exhibit varying degrees of site-fidelity to the previous year's territory or breeding area, a phenomenon we refer to as incomplete breeding site-fidelity. If the territory they occupy is located beyond the bounds of the study area or search area (i.e., they have emigrated from the study area), the bird will go undetected and is therefore indistinguishable from dead individuals in capture-mark-recapture studies. Differential emigration rates confound inferences regarding differences in survival between sexes and among species if apparent survival rates are used as estimates of true survival. Moreover, the bias introduced by using apparent survival rates for true survival rates can have profound effects on the predictions of population persistence through time, source/sink dynamics, and other aspects of life-history theory. We investigated four study design and analysis approaches that result in apparent survival estimates that are closer to true survival estimates. Our motivation for this research stemmed from a multi-year capture-recapture study of Prothonotary Warblers (Protonotaria citrea) on multiple study plots within a larger landscape of suitable breeding habitat where substantial inter-annual movements of marked individuals among neighboring study plots was documented. We wished to quantify the effects of this type of movement on annual survival estimation. The first two study designs we investigated involved marking birds in a core area and resighting them in the core as well as an area surrounding the core. For the first of these two designs, we demonstrated that as the resighting area surrounding the core gets progressively larger, and more "emigrants" are resighted, apparent survival estimates begin to approximate true survival rates (bias < 0.01). However, given observed inter-annual movements of birds, it is likely to be logistically impractical to resight birds on sufficiently large surrounding areas to minimize bias. Therefore

  11. HotSpot Wizard 2.0: automated design of site-specific mutations and smart libraries in protein engineering.

    PubMed

    Bendl, Jaroslav; Stourac, Jan; Sebestova, Eva; Vavra, Ondrej; Musil, Milos; Brezovsky, Jan; Damborsky, Jiri

    2016-07-08

    HotSpot Wizard 2.0 is a web server for automated identification of hot spots and design of smart libraries for engineering proteins' stability, catalytic activity, substrate specificity and enantioselectivity. The server integrates sequence, structural and evolutionary information obtained from 3 databases and 20 computational tools. Users are guided through the processes of selecting hot spots using four different protein engineering strategies and optimizing the resulting library's size by narrowing down a set of substitutions at individual randomized positions. The only required input is a query protein structure. The results of the calculations are mapped onto the protein's structure and visualized with a JSmol applet. HotSpot Wizard lists annotated residues suitable for mutagenesis and can automatically design appropriate codons for each implemented strategy. Overall, HotSpot Wizard provides comprehensive annotations of protein structures and assists protein engineers with the rational design of site-specific mutations and focused libraries. It is freely available at http://loschmidt.chemi.muni.cz/hotspotwizard. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

  12. HotSpot Wizard 2.0: automated design of site-specific mutations and smart libraries in protein engineering

    PubMed Central

    Bendl, Jaroslav; Stourac, Jan; Sebestova, Eva; Vavra, Ondrej; Musil, Milos; Brezovsky, Jan; Damborsky, Jiri

    2016-01-01

    HotSpot Wizard 2.0 is a web server for automated identification of hot spots and design of smart libraries for engineering proteins’ stability, catalytic activity, substrate specificity and enantioselectivity. The server integrates sequence, structural and evolutionary information obtained from 3 databases and 20 computational tools. Users are guided through the processes of selecting hot spots using four different protein engineering strategies and optimizing the resulting library's size by narrowing down a set of substitutions at individual randomized positions. The only required input is a query protein structure. The results of the calculations are mapped onto the protein's structure and visualized with a JSmol applet. HotSpot Wizard lists annotated residues suitable for mutagenesis and can automatically design appropriate codons for each implemented strategy. Overall, HotSpot Wizard provides comprehensive annotations of protein structures and assists protein engineers with the rational design of site-specific mutations and focused libraries. It is freely available at http://loschmidt.chemi.muni.cz/hotspotwizard. PMID:27174934

  13. Balancing Energy, Food Security, and Critical Ecosystems: Dam Siting, Design and Operations in the Mekong River Basin

    NASA Astrophysics Data System (ADS)

    Wild, T. B.; Reed, P. M.; Loucks, D.

    2016-12-01

    The Mekong River basin in Southeast Asia is one of several river basins with exceptionally high biodiversity value where intensive hydropower dam development is anticipated. In the Mekong basin, over 100 dams are planned to be constructed in the next 20-30 years. As planned, these dams will alter the river's natural water, sediment and nutrient flows, and will fragment fish migration pathways. In doing so, they will degrade one of the world's most productive freshwater fish habitats, upon which some 60 million people depend for food and income security. For those dams that have not yet been constructed, there still exist opportunities to modify their siting, design and operation (SDO) to achieve a more balanced set of tradeoffs among hydropower production, sediment/nutrient passage and adult fish/larvae passage. We present a successful case study wherein we explored such alternative SDO opportunities in partnership with the Government of Cambodia for Sambor Dam, planned to be built on the main stem of the Mekong. Sambor would be one of the world's longest dams, spanning 18 km across the river just upstream of (1) Tonle Sap Lake, which supplies 70% of Cambodians' protein, and (2) the Mekong Delta in Vietnam, responsible for 50% of Vietnam's rice production. We will describe key dam siting and design modifications required to mitigate ecological impacts. We will then focus on the most promising alternative dam siting/design concept, exploring the reservoir operations space to demonstrate that a complex set of tradeoffs exist among a diverse set of energy and ecosystem objectives. Results indicate that even for a hydrologically small reservoir, a wide array of potential reservoir operating policies exist that have vastly different food-energy implications. While some policies would significantly mitigate ecological impacts, many policies exist that would pose a severe threat to the sustainability of the fishery. Failure to sample the reservoir operations space at

  14. A novel approach for determination of fundamental physical transport processes in natural channel design restoration sites with river steering structures

    NASA Astrophysics Data System (ADS)

    Robinson, J.; Endreny, T. A.; Becker, J. F.; Kroll, C.

    2012-12-01

    River restoration projects in the United States are frequently proposed and constructed with the intention of improving water quality, yet relatively little evidence exists regarding the success of these efforts. Many projects use an approach known as natural channel design (NCD), and include river steering structures. Prior assessment of water quality improvements within NCD sites has involved hydrologic retention modeling using a non-reactive tracer, with the goal of separately identifying hyporheic and surface transient storage (STS). A comparative approach involving NCD and non-NCD sites used by the authors yielded mixed results: although physically-based assessments of STS profiles in many NCD sites support larger STS zones than non-NCD sites, these differences are not apparent when examining common transient storage metrics. Inverse modeling within nine NCD sites reveals additional obstacles, including generation of spurious lateral inflow/outflow values, limited detection of hyporheic processes due to strong surface transient storage, shear and Taylor dispersion, and divergent temporal patterns of solute flux over channel cross sections bounding structures. To overcome the obstacles encountered with 1D inverse modeling, data is presented from a new approach used in NCD reaches. This approach involves deriving a mass flux signature via pairing velocity and channel geometry with multiple electrical conductivity (EC) loggers deployed laterally at control cross sections (CCS). These CCS bound sub-reach segments (15 total across four NCD reaches) that include river steering structures and intermediate geomorphic features. Velocity and geometry measurements yield discharge values surrounding each EC logger which are used to weight a composite mass flux breakthrough curve above, within, and below each segment. Composite mass flux signatures reflect exchange processes that are not fully integrated laterally immediately below structures, and can be analyzed via

  15. Site-specifically modified oligodeoxynucleotides as probes for the structural and biological effect of DNA-damaging agents

    SciTech Connect

    Basu, A.K.; Essigmann, J.M.

    1988-01-01

    This review critically analyzes the state of knowledge on the preparation, characterization, and uses of site-specifically modified DNA segments. Although these substrates have begun to have an impact upon several fields, the review focuses upon their applications in site-directed mutagenesis studies and for defining the effect of chemical damage upon DNA structure. Oligonucleotides have been synthesized containing alkylated DNA bases, aromatic amine adducts, base oxidation products, cyclic nucleic acid adducts, model apurinic/apyrimidinic sites, UV and psoralen photoadducts, and several antitumor drug-DNA covalent complexes. Below, the authors shall describe the progress to date on synthesis of site-specifically modified DNA segments and how these oligonucleotides have been used to further their understanding of the roles of individual DNA adducts in toxicology. The structures of the DNA adducts and adduct-derived products discussed in this review are presented. 168 references.

  16. Remedial action and site design for stabilization of the inactive uranium mill tailings sites at Slick Rock, Colorado. Attachment 2, Geology report

    SciTech Connect

    Not Available

    1993-07-01

    This report presents geologic considerations that are pertinent to the Remedial Action Plan for Slick Rock mill tailings. Topics covered include regional geology, site geology, geologic stability, and geologic suitability.

  17. Remedial Action Plan and site design for stabilization of the inactive uranium mill tailings site at Durango, Colorado: Attachment 3, Groundwater hydrology report. Revised final report

    SciTech Connect

    Not Available

    1991-12-01

    The US Environmental Protection Agency (EPA) has established health and environmental protection regulations to correct and prevent groundwater contamination resulting from processing activities at inactive uranium milling sites. According to the Uranium Mill Tailings Radiation Control Act of 1978, (UMTRCA) the US Department of Energy (DOE) is responsible for assessing the inactive uranium processing sites. The DOE has determined this assessment shall include information on hydrogeologic site characterization. The water resources protection strategy that describes how the proposed action will comply with the EPA groundwater protection standards is presented in Attachment 4. Site characterization activities discussed in this section include: Characterization of the hydrogeologic environment; characterization of existing groundwater quality; definition of physical and chemical characteristics of the potential contaminant source; and description of local water resources.

  18. Comment and response document on the final remedial action plan and site design for stabilization of the inactive Uranium Mill Tailings Site at Lakeview, Oregon

    SciTech Connect

    Not Available

    1991-10-01

    This report contains comments provided by the Oregon Department of Energy and responses to these comments on the final remedial action plan for the inactive uranium mill tailings site at Lakeview, Oregon.

  19. Volunteers Oriented Interface Design for the Remote Navigation of Rescue Robots at Large-Scale Disaster Sites

    NASA Astrophysics Data System (ADS)

    Yang, Zhixiao; Ito, Kazuyuki; Saijo, Kazuhiko; Hirotsune, Kazuyuki; Gofuku, Akio; Matsuno, Fumitoshi

    This paper aims at constructing an efficient interface being similar to those widely used in human daily life, to fulfill the need of many volunteer rescuers operating rescue robots at large-scale disaster sites. The developed system includes a force feedback steering wheel interface and an artificial neural network (ANN) based mouse-screen interface. The former consists of a force feedback steering control and a six monitors’ wall. It provides a manual operation like driving cars to navigate a rescue robot. The latter consists of a mouse and a camera’s view displayed in a monitor. It provides a semi-autonomous operation by mouse clicking to navigate a rescue robot. Results of experiments show that a novice volunteer can skillfully navigate a tank rescue robot through both interfaces after 20 to 30 minutes of learning their operation respectively. The steering wheel interface has high navigating speed in open areas, without restriction of terrains and surface conditions of a disaster site. The mouse-screen interface is good at exact navigation in complex structures, while bringing little tension to operators. The two interfaces are designed to switch into each other at any time to provide a combined efficient navigation method.

  20. Design, synthesis and antiviral activity of entry inhibitors that target the CD4-binding site of HIV-1

    PubMed Central

    Curreli, Francesca; Choudhury, Spreeha; Pyatkin, Ilya; Zagorodnikov, Victor P.; Bulay, Anna Khulianova; Altieri, Andrea; Kwon, Young Do; Kwong, Peter D.; Debnath, Asim K.

    2012-01-01

    The CD4 binding site on HIV-1 gp120 has been validated as a drug target to prevent HIV-1 entry to cells. Previously, we identified two small molecule inhibitors consisting of a 2,2,6,6-tetramethylpiperidine ring linked by an oxalamide to a p-halide-substituted phenyl group, which target this site, specifically, a cavity termed “Phe43 cavity”. Here we use synthetic chemistry, functional assessment and structure-based analysis to explore variants of each region of these inhibitors for improved antiviral properties. Alterations of the phenyl group and of the oxalamide linker indicated that these regions were close to optimal in the original lead compounds. Design of a series of compounds, where the tetramethylpiperidine ring was replaced with new scaffolds, lead to improved antiviral activity. These new scaffolds provide insight into the surface chemistry at the entrance of the cavity and offer additional opportunities by which to optimize further these potential-next-generation therapeutics and microbicides against HIV-1. PMID:22524483

  1. On the Proper Estimation of the Confidence Interval for the Design Formula of Blast-Induced Vibrations with Site Records

    NASA Astrophysics Data System (ADS)

    Yan, W. M.; Yuen, Ka-Veng

    2015-01-01

    Blast-induced ground vibration has received much engineering and public attention. The vibration is often represented by the peak particle velocity (PPV) and the empirical approach is employed to describe the relationship between the PPV and the scaled distance. Different statistical methods are often used to obtain the confidence level of the prediction. With a known scaled distance, the amount of explosives in a planned blast can then be determined by a blast engineer when the PPV limit and the confidence level of the vibration magnitude are specified. This paper shows that these current approaches do not incorporate the posterior uncertainty of the fitting coefficients. In order to resolve this problem, a Bayesian method is proposed to derive the site-specific fitting coefficients based on a small amount of data collected at an early stage of a blasting project. More importantly, uncertainty of both the fitting coefficients and the design formula can be quantified. Data collected from a site formation project in Hong Kong is used to illustrate the performance of the proposed method. It is shown that the proposed method resolves the underestimation problem in one of the conventional approaches. The proposed approach can be easily conducted using spreadsheet calculation without the need for any additional tools, so it will be particularly welcome by practicing engineers.

  2. Design of a carbonic anhydrase IX active-site mimic to screen inhibitors for possible anticancer properties.

    PubMed

    Genis, Caroli; Sippel, Katherine H; Case, Nicolette; Cao, Wengang; Avvaru, Balendu Sankara; Tartaglia, Lawrence J; Govindasamy, Lakshmanan; Tu, Chingkuang; Agbandje-McKenna, Mavis; Silverman, David N; Rosser, Charles J; McKenna, Robert

    2009-02-17

    Recently, a convincing body of evidence has accumulated suggesting that the overexpression of carbonic anhydrase isozyme IX (CA IX) in some cancers contributes to the acidification of the extracellular matrix, which in turn promotes the growth and metastasis of the tumor. These observations have made CA IX an attractive drug target for the selective treatment of certain cancers. Currently, there is no available X-ray crystal structure of CA IX, and this lack of availability has hampered the rational design of selective CA IX inhibitors. In light of these observations and on the basis of structural alignment homology, using the crystal structure of carbonic anhydrase II (CA II) and the sequence of CA IX, a double mutant of CA II with Ala65 replaced by Ser and Asn67 replaced by Gln has been constructed to resemble the active site of CA IX. This CA IX mimic has been characterized kinetically using (18)O-exchange and structurally using X-ray crystallography, alone and in complex with five CA sulfonamide-based inhibitors (acetazolamide, benzolamide, chlorzolamide, ethoxzolamide, and methazolamide), and compared to CA II. This structural information has been evaluated by both inhibition studies and in vitro cytotoxicity assays and shows a correlated structure-activity relationship. Kinetic and structural studies of CA II and CA IX mimic reveal chlorzolamide to be a more potent inhibitor of CA IX, inducing an active-site conformational change upon binding. Additionally, chlorzolamide appears to be cytotoxic to prostate cancer cells. This preliminary study demonstrates that the CA IX mimic may provide a useful model to design more isozyme-specific CA IX inhibitors, which may lead to development of new therapeutic treatments of some cancers.

  3. sgRNAcas9: a software package for designing CRISPR sgRNA and evaluating potential off-target cleavage sites.

    PubMed

    Xie, Shengsong; Shen, Bin; Zhang, Chaobao; Huang, Xingxu; Zhang, Yonglian

    2014-01-01

    Although the CRISPR/Cas9/sgRNA system efficiently cleaves intracellular DNA at desired target sites, major concerns remain on potential "off-target" cleavage that may occur throughout the whole genome. In order to improve CRISPR-Cas9 specificity for targeted genome editing and transcriptional control, we describe a bioinformatics tool "sgRNAcas9", which is a software package developed for fast design of CRISPR sgRNA with minimized off-target effects. This package consists of programs to perform a search for CRISPR target sites (protospacers) with user-defined parameters, predict genome-wide Cas9 potential off-target cleavage sites (POT), classify the POT into three categories, batch-design oligonucleotides for constructing 20-nt (nucleotides) or truncated sgRNA expression vectors, extract desired length nucleotide sequences flanking the on- or off-target cleavage sites for designing PCR primer pairs to validate the mutations by T7E1 cleavage assay. Importantly, by identifying potential off-target sites in silico, the sgRNAcas9 allows the selection of more specific target sites and aids the identification of bona fide off-target sites, significantly facilitating the design of sgRNA for genome editing applications. sgRNAcas9 software package is publicly available at BiooTools website (www.biootools.com) under the terms of the GNU General Public License.

  4. REVIEW REPORT: BUILDING C-400 THERMAL TREATMENT 90 PERCENT REMEDIAL DESIGN REPORT AND SITE INVESTIGATION, PGDP, PADUCAH, KENTUCKY

    SciTech Connect

    Looney, B; Jed Costanza, J; Eva Davis, E; Joe Rossabi, J; Lloyd Stewart, L; Hans Stroo, H

    2007-08-15

    On 9 April 2007, the U.S. Department of Energy (DOE) Headquarters, Office of Soil and Groundwater Remediation (EM-22) initiated an Independent Technical Review (ITR) of the 90% Remedial Design Report (RDR) and Site Investigation (RDSI) for thermal treatment of trichloroethylene (TCE) in the soil and groundwater in the vicinity of Building C-400 at the Paducah Gaseous Diffusion Plant (PGDP). The general ITR goals were to assess the technical adequacy of the 90% RDSI and provide recommendations sufficient for DOE to determine if modifications are warranted pertaining to the design, schedule, or cost of implementing the proposed design. The ultimate goal of the effort was to assist the DOE Paducah/Portsmouth Project Office (PPPO) and their contractor team in ''removing'' the TCE source zone located near the C-400 Building. This report provides the ITR findings and recommendations and supporting evaluations as needed to facilitate use of the recommendations. The ITR team supports the remedial action objective (RAO) at C-400 to reduce the TCE source area via subsurface Electrical Resistance Heating (ERH). Further, the ITR team commends PPPO, their contractor team, regulators, and stakeholders for the significant efforts taken in preparing the 90% RDR. To maximize TCE removal at the target source area, several themes emerge from the review which the ITR team believes should be considered and addressed before implementing the thermal treatment. These themes include the need for: (1) Accurate and site-specific models as the basis to verify the ERH design for full-scale implementation for this challenging hydrogeologic setting; (2) Flexible project implementation and operation to allow the project team to respond to observations and data collected during construction and operation; (3) Defensible performance metrics and monitoring, appropriate for ERH, to ensure sufficient and efficient clean-up; and (4) Comprehensive (creative and diverse) contingencies to address the

  5. Investigation of the histamine H3 receptor binding site. Design and synthesis of hybrid agonists with a lipophilic side chain.

    PubMed

    Ishikawa, Makoto; Watanabe, Takashi; Kudo, Toshiaki; Yokoyama, Fumikazu; Yamauchi, Miki; Kato, Kazuhiko; Kakui, Nobukazu; Sato, Yasuo

    2010-09-09

    As a part of our search for novel histamine H3 receptor agonists, we designed and synthesized hybrid compounds in which the lipophilic (4'-alkylphenylthio)ethyl moiety of a novel H3 receptor agonist, 4-(2-(4'-tert-butylphenylthio)ethyl)-1H-imidazole (1), was incorporated into N(alpha)-methylhistamine, immepip, and immethridine derivatives. These hybrid compounds were expected to interact concurrently with the histamine-binding site and a putative hydrophobic region in the H3 receptor. Among them, piperidine- and pyridine-type derivatives displayed partial agonist activity, and (S)-4-(1-(1H-imidazol-4-yl)-2-(4-(trifluoromethyl)phenylthio)ethyl)piperidine (36) was identified as a potent H3 agonist. We performed computational docking studies to examine the binding mode of the agonists. The results indicated that immepip interacts with the key residues, Asp114 and Glu206, in a different manner from histamine. The binding mode of 36 to these residues is similar to that of immepip, and the lipophilic tail of 36 has an additional interaction with a hydrophobic region in transmembrane helix 6 of the receptor. These results indicated that 36 served as a useful tool for studies on receptor-agonist interactions and drug design.

  6. Use of Mathematical Models in the Design and Performance Evaluation of a Surfactant Flushing Demonstration at the Bachman Road Site

    NASA Astrophysics Data System (ADS)

    Abriola, L. M.; Drummond, C. D.; Lemke, L. D.; Rathfelder, K. M.; Pennell, K. D.

    2001-05-01

    This presentation provides an overview of the design and performance evaluation of a surfactant enhanced remediation pilot demonstration conducted in the summer of 2000 at a former dry cleaning facility in Oscoda, Michigan, USA. The unconfined contaminated formation is composed of relatively homogeneous glacial outwash sands, underlain by a thick clay layer. Core samples have revealed the presence of a reasonably persistent coarse sand and gravel layer at a depth of 11-16 feet and a sand/silt/clay transition zone at the base of the aquifer. A narrow tetrachloroethylene (PCE) plume emanates from the suspected source area, beneath the former dry cleaning building, and discharges into Lake Huron, approximately 700 feet down gradient. There is little evidence of microbial plume attenuation at the site. Aqueous samples from multilevel piezometers installed beneath the building have confirmed the presence of residual PCE within the coarse sand and gravel layer and have detected consistently high PCE concentrations at the base of the aquifer. The actual distribution and volume of entrapped PCE, however, is unknown. A surfactant injection and recovery scheme was designed and implemented to effectively flush the identified source area beneath the building. In this scheme, a line of water injection wells was installed behind the surfactant injection points to control surfactant delivery and maximize solubilized plume capture. Prior to surfactant injection, conservative and partitioning tracer tests were also conducted to confirm sweep and estimate source zone mass. Mass recovery calculations indicate that more than 94% of the injected surfactant and approximately 19 liters of PCE were recovered during the test. This volume of DNAPL is consistent with estimated low saturations within the swept zone. Single and multiphase transport models were employed to aid in remedial design and predict system performance. For the model simulations, input parameters were determined from

  7. Hyperglycosylated Stable Core Immunogens Designed To Present the CD4 Binding Site Are Preferentially Recognized by Broadly Neutralizing Antibodies

    PubMed Central

    Ingale, Jidnyasa; Tran, Karen; Kong, Leopold; Dey, Barna; McKee, Krisha; Schief, William; Kwong, Peter D.; Mascola, John R.

    2014-01-01

    ABSTRACT The HIV-1 surface envelope glycoprotein (Env) trimer mediates entry into CD4+ CCR5+ host cells. Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. Env also contains variable regions and protein surfaces occluded within the trimer that elicit nonneutralizing antibodies. Here we engineered additional N-linked glycans onto a cysteine-stabilized gp120 core (0G) deleted of its major variable regions to preferentially expose the conformationally fixed CD4bs. Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. Following purification, most glycoproteins, except for 10G, were recognized by broadly neutralizing CD4bs-directed antibodies. Gel and glycan mass spectrometry confirmed that additional N-glycans were posttranslationally added to the redesigned cores. Binding kinetics revealed high-affinity recognition by seven broadly neutralizing CD4bs-directed antibodies and low to no binding by non-broadly neutralizing CD4bs-directed antibodies. Rabbits inoculated with the hyperglycosylated cores elicited IgM and IgG responses to each given protein that were similar in their neutralization characteristics to those elicited by parental 0G. Site-specific glycan masking effects were detected in the elicited sera, and the antisera competed with b12 for CD4bs-directed binding specificity. However, the core-elicited sera showed limited neutralization activity. Trimer priming or boosting of the core immunogens elicited tier 1-level neutralization that mapped to both the CD4bs and V3 and appeared to be trimer dependent. Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. IMPORTANCE

  8. Remedial Action Plan and site design for stabilization of the inactive uranium mill tailings site at Gunnison, Colorado. Attachment 4, Water resources protection strategy: Final report

    SciTech Connect

    Not Available

    1992-10-01

    To achieve compliance with the proposed US Environmental Protection Agency (EPA) groundwater protection standards the US Department of Energy (DOE) proposes to meet background concentrations or the EPA maximum concentration limits (MCLS) for hazardous constituents in groundwater in the uppermost aquifer at the point of compliance (POC) at the Gunnison Uranium Mill Tailings Remedial Action (UMTRA) Project disposal site near Gunnison, Colorado. The proposed remedial action will ensure protection of human health and the environment. A summary of the principal features of the water resources protection strategy for the Gunnison disposal site is included in this report.

  9. Site-Directed Mutagenesis of a Hyperthermophilic Endoglucanase Cel12B from Thermotoga maritima Based on Rational Design

    PubMed Central

    Zhang, Jinfeng; Shi, Hao; Xu, Linyu; Zhu, Xiaoyan; Li, Xiangqian

    2015-01-01

    To meet the demand for the application of high activity and thermostable cellulases in the production of new-generation bioethanol from nongrain-cellulose sources, a hyperthermostable β-1,4-endoglucase Cel12B from Thermotoga maritima was selected for further modification by gene site-directed mutagenesis method in the present study, based on homology modeling and rational design. As a result, two recombinant enzymes showed significant improvement in enzyme activity by 77% and 87%, respectively, higher than the parental enzyme TmCel12B. Furthermore, the two mutants could retain 80% and 90.5% of their initial activity after incubation at 80°C for 8 h, while only 45% for 5 h to TmCel12B. The Km and Vmax of the two recombinant enzymes were 1.97±0.05 mM, 4.23±0.15 μmol·mg-1·min-1 of TmCel12B-E225H-K207G-D37V, and 2.97±0.12 mM, 3.15±0.21 μmol·mg-1·min-1 of TmCel12B-E225H-K207G, respectively, when using CMC-Na as the substrate. The roles of the mutation sites were also analyzed and evaluated in terms of electron density, hydrophobicity of the modeled protein structures. The recombinant enzymes may be used in the hydrolysis of cellulose at higher temperature in the future. It was concluded that the gene mutagenesis approach of a certain active residues may effectively improve the performance of cellulases for the industrial applications and contribute to the study the thermostable mechanism of thermophilic enzymes. PMID:26218520

  10. Rational Design of Fatty Acid Amide Hydrolase Inhibitors that Act by Covalently Bonding to Two Active Site Residues

    PubMed Central

    Otrubova, Katerina; Brown, Monica; McCormick, Michael S.; Han, Gye W.; O’Neal, Scott T.; Cravatt, Benjamin F.; Stevens, Raymond C.; Lichtman, Aron H.; Boger, Dale L.

    2013-01-01

    The design and characterization of α-ketoheterocycle fatty acid amide hydrolase (FAAH) inhibitors are disclosed that additionally and irreversibly target a cysteine (Cys269) found in the enzyme cytosolic port while maintaining the reversible covalent Ser241 attachment responsible for their rapid and initially reversible enzyme inhibition. Two α-ketooxazoles (3 and 4) containing strategically placed electrophiles at the C5 position of the pyridyl substituent of 2 (OL-135) were prepared and examined as inhibitors of FAAH. Consistent with the observed time-dependent non-competitive inhibition, the co-crystal X-ray structure of 3 bound to a humanized variant of rat FAAH revealed that 3 was not only covalently bound to the active site catalytic nucleophile Ser241 as a deprotonated hemiketal, but also to Cys269 through the pyridyl C5-substituent, thus providing an inhibitor with dual covalent attachment in the enzyme active site. In vivo characterization of the prototypical inhibitors in mice demonstrate that they raise endogenous brain levels of FAAH substrates to a greater extent and for a much longer duration (>6 h) than the reversible inhibitor 2, indicating that the inhibitors accumulate and persist in the brain to completely inhibit FAAH for a prolonged period. Consistent with this behavior and the targeted irreversible enzyme inhibition, 3 reversed cold allodynia in the chronic constriction injury model of neuropathic pain in mice for a sustained period (>6 h) beyond that observed with the reversible inhibitor 2, providing effects that were unchanged over the 1–6 h time course monitored. PMID:23581831

  11. Site-specific non-LTR retrotransposons.

    PubMed

    Fujiwara, Haruhiko

    2015-04-01

    Although most of non-long terminal repeat (non-LTR) retrotransposons are incorporated in the host genome almost randomly, some non-LTR retrotransposons are incorporated into specific sequences within a target site. On the basis of structural and phylogenetic features, non-LTR retrotransposons are classified into two large groups, restriction enzyme-like endonuclease (RLE)-encoding elements and apurinic/apyrimidinic endonuclease (APE)-encoding elements. All clades of RLE-encoding non-LTR retrotransposons include site-specific elements. However, only two of more than 20 APE-encoding clades, Tx1 and R1, contain site-specific non-LTR elements. Site-specific non-LTR retrotransposons usually target within multi-copy RNA genes, such as rRNA gene (rDNA) clusters, or repetitive genomic sequences, such as telomeric repeats; this behavior may be a symbiotic strategy to reduce the damage to the host genome. Site- and sequence-specificity are variable even among closely related non-LTR elements and appeared to have changed during evolution. In the APE-encoding elements, the primary determinant of the sequence- specific integration is APE itself, which nicks one strand of the target DNA during the initiation of target primed reverse transcription (TPRT). However, other factors, such as interaction between mRNA and the target DNA, and access to the target region in the nuclei also affect the sequence-specificity. In contrast, in the RLE-encoding elements, DNA-binding motifs appear to affect their sequence-specificity, rather than the RLE domain itself. Highly specific integration properties of these site-specific non-LTR elements make them ideal alternative tools for sequence-specific gene delivery, particularly for therapeutic purposes in human diseases.

  12. Theoretical and experimental design of site-specific applicators and heating protocols for interstitial ultrasound thermal therapy

    NASA Astrophysics Data System (ADS)

    Tyreus, Per Daniel; Nau, William H.; Wu, Alex; Diederich, Chris J.

    2003-06-01

    Theoretical and experimental approaches were used to develop and evaluate site-specific designs of internally cooled direct coupled (ICDC) and catheter-cooled (CC) ultrasound applicators for thermal coagulation of disease in the prostate, liver, brain, and uterus. The diameter of an interstitial applicator can influence its clinical practicality and effectiveness as well as application site. One purpose of this study was to determine whether the use of larger ultrasound transducers and the inherent increase in applicator size could be justified by potentially producing larger lesion diameters. A second purpose was to explore how the response of tissue acoustic attenuation to heating effects lesion size and preferred applicator configuration. Four applicator configurations and sizes were studied using ex vivo tissue experiments in liver and beef and using acoustic and biothermal simulations. Transmission attenuation measurements showed a 6 to 8 fold increase in baseline tissue attenution inside interstitial ultrasound lesions. Formation of these high attenuation zones in lesions reduced potential lesion size. Larger applicators produced lesions with radial penetration depths superior to their smaller counterparts at power levels in the 20-40W /cm range. The higher cooling rates along the outer surface of the larger diameter applicators due to their greater surface area was a dominant factor in increasing lesion size. The higher cooling rates pushed the maximum temperature farther from the applicator surface and reduced the formation of high acoustic attenuation tissue zones. Acoustic and biothermal simulations matched the experimental data well and were applied to model these applicators within sites of clinical interest such as prostate, uterine fibroid, brain, and normal liver. Lesions of 3.9 to 4.7cm diameter were predicted for moderately perfused tissues such as prostate and fibroid and 2.8 to 3.2cm for highly perfused tissues such as normal liver. Feedback

  13. Tunable, Chemo- and Site-Selective Nitrene Transfer Reactions through the Rational Design of Silver(I) Catalysts.

    PubMed

    Alderson, Juliet M; Corbin, Joshua R; Schomaker, Jennifer M

    2017-08-08

    Carbon-nitrogen (C-N) bonds are ubiquitous in pharmaceuticals, agrochemicals, diverse bioactive natural products, and ligands for transition metal catalysts. An effective strategy for introducing a new C-N bond into a molecule is through transition metal-catalyzed nitrene transfer chemistry. In these reactions, a metal-supported nitrene can either add across a C═C bond to form an aziridine or insert into a C-H bond to furnish the corresponding amine. Typical catalysts for nitrene transfer include Rh2Ln and Ru2Ln complexes supported by bridging carboxylate and related ligands, as well as complexes based on Cu, Co, Ir, Fe, and Mn supported by porphyrins and related ligands. A limitation of metal-catalyzed nitrene transfer is the ability to predictably select which specific site will undergo amination in the presence of multiple reactive groups; thus, many reactions rely primarily on substrate control. Achieving true catalyst-control over nitrene transfer would open up exciting possibilities for flexible installation of new C-N bonds into hydrocarbons, natural product-inspired scaffolds, existing pharmaceuticals or biorenewable building blocks. Silver-catalyzed nitrene transfer enables flexible control over the position at which a new C-N bond is introduced. Ag(I) supported by simple N-donor ligands accommodates a diverse range of coordination geometries, from linear to tetrahedral to seesaw, enabling the electronic and steric parameters of the catalyst to be tuned independently. In addition, the ligand, Ag salt counteranion, Ag/ligand ratio and the solvent all influence the fluxional and dynamic behavior of Ag(I) complexes in solution. Understanding the interplay of these parameters to manipulate the behavior of Ag-nitrenes in a predictable manner is a key design feature of our work. In this Account, we describe successful applications of a variety of design principles to tunable, Ag-catalyzed aminations, including (1) changing Ag/ligand ratios to influence

  14. A Multi-site, Two-Phase, Prescription Opioid Addiction Treatment Study (POATS): Rationale, Design, and Methodology

    PubMed Central

    Weiss, Roger D.; Potter, Jennifer Sharpe; Provost, Scott E.; Huang, Zhen; Jacobs, Petra; Hasson, Albert; Lindblad, Robert; Connery, Hilary Smith; Prather, Kristi; Ling, Walter

    2010-01-01

    The National Institute on Drug Abuse Clinical Trials Network launched the Prescription Opioid Addiction Treatment Study (POATS) in response to rising rates of prescription opioid dependence and gaps in understanding the optimal course of treatment for this population. POATS employed a multi-site, two-phase adaptive, sequential treatment design to approximate clinical practice. The study took place at 10 community treatment programs around the United States. Participants included men and women age ≥18 who met Diagnostic and Statistical Manual, 4th Edition criteria for dependence upon prescription opioids, with physiologic features; those with a prominent history of heroin use (according to pre-specified criteria) were excluded. All participants received buprenorphine/naloxone (bup/nx). Phase 1 consisted of 4 weeks of bup/nx treatment, including a 14-day dose taper, with 8 weeks of follow-up. Phase 1 participants were monitored for treatment response during these 12 weeks. Those who relapsed to opioid use, as defined by pre-specified criteria, were invited to enter Phase 2; Phase 2 consisted of 12 weeks of bup/nx stabilization treatment, followed by a 4-week taper and 8 weeks of post-treatment follow-up. Participants were randomized at the beginning of Phase 1 to receive bup/nx, paired with either Standard Medical Management (SMM) or Enhanced Medical Management (EMM; defined as SMM plus individual drug counseling). Eligible participants entering Phase 2 were re-randomized to either EMM or SMM. POATS was developed to determine what benefit, if any, EMM offers over SMM in short-term and longer-term treatment paradigm. This paper describes the rationale and design of the study. PMID:20116457

  15. 75 FR 38129 - Freescale Semiconductor, Inc., Hardware/Software Design and Manufacturing A Including On-Site...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-01

    ... Design, Design Solutions, Inc., Veriseo, SiliconElite and MicroLogic, Inc., Austin, TX; Amended..., GDA Technologies, Inc., Manpower, Ion Design, Design Solutions, Inc., Veriseo, SilconElite and Micro... Technologies, Inc., Manpower, Ion Design, Design Solutions, Inc., Veriseo, SiliconElite and MicroLogic,...

  16. Conserving and Enhancing the Natural Environment: A Guide for Planning, Design, Construction, and Maintenance on New & Existing School Sites.

    ERIC Educational Resources Information Center

    Maryland State Dept. of Education, Baltimore. School Facilities Branch.

    Natural environments on school sites provide considerable multi-disciplinary educational opportunities, many of which are "hands-on" experiences that stimulate learning. This document presents guidelines on conserving and enhancing the natural environment on school sites. It provides guidance for developing the site requirements in…

  17. Computerized stratified random site-selection approaches for design of a ground-water-quality sampling network

    USGS Publications Warehouse

    Scott, J.C.

    1990-01-01

    Computer software was written to randomly select sites for a ground-water-quality sampling network. The software uses digital cartographic techniques and subroutines from a proprietary geographic information system. The report presents the approaches, computer software, and sample applications. It is often desirable to collect ground-water-quality samples from various areas in a study region that have different values of a spatial characteristic, such as land-use or hydrogeologic setting. A stratified network can be used for testing hypotheses about relations between spatial characteristics and water quality, or for calculating statistical descriptions of water-quality data that account for variations that correspond to the spatial characteristic. In the software described, a study region is subdivided into areal subsets that have a common spatial characteristic to stratify the population into several categories from which sampling sites are selected. Different numbers of sites may be selected from each category of areal subsets. A population of potential sampling sites may be defined by either specifying a fixed population of existing sites, or by preparing an equally spaced population of potential sites. In either case, each site is identified with a single category, depending on the value of the spatial characteristic of the areal subset in which the site is located. Sites are selected from one category at a time. One of two approaches may be used to select sites. Sites may be selected randomly, or the areal subsets in the category can be grouped into cells and sites selected randomly from each cell.

  18. A Hierarchical Modeling Approach to Data Analysis and Study Design in a Multi-Site Experimental fMRI Study

    ERIC Educational Resources Information Center

    Zhou, Bo; Konstorum, Anna; Duong, Thao; Tieu, Kinh H.; Wells, William M.; Brown, Gregory G.; Stern, Hal S.; Shahbaba, Babak

    2013-01-01

    We propose a hierarchical Bayesian model for analyzing multi-site experimental fMRI studies. Our method takes the hierarchical structure of the data (subjects are nested within sites, and there are multiple observations per subject) into account and allows for modeling between-site variation. Using posterior predictive model checking and model…

  19. 77 FR 55144 - Ocean Dumping; Designation of Ocean Dredged Material Disposal Sites Offshore of Yaquina Bay, Oregon

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-07

    ... higher cost of monitoring sites in deeper waters and further offshore. Historic disposal has occurred at... continental shelf and other such sites where historical disposal has occurred (40 CFR 228.5(e)). Disposal... Proximity to the Site of any Significant Natural or Cultural Feature of Historical Importance (40 CFR...

  20. Detailed computational study of the active site of the hepatitis C viral RNA polymerase to aid novel drug design.

    PubMed

    Barakat, Khaled H; Law, John; Prunotto, Alessio; Magee, Wendy C; Evans, David H; Tyrrell, D Lorne; Tuszynski, Jack; Houghton, Michael

    2013-11-25

    The hepatitis C virus (HCV) RNA polymerase, NS5B, is a leading target for novel and selective HCV drug design. The enzyme has been the subject of intensive drug discovery aimed at developing direct acting antiviral (DAA) agents that inhibit its activity and hence prevent the virus from replicating its genome. In this study, we focus on one class of NS5B inhibitors, namely nucleos(t)ide mimetics. Forty-one distinct nucleotide structures have been modeled within the active site of NS5B for the six major HCV genotypes. Our comprehensive modeling protocol employed 287 different molecular dynamics simulations combined with the molecular mechanics/Poisson-Boltzmann surface area (MM-PBSA) methodology to rank and analyze these structures for all genotypes. The binding interactions of the individual compounds have been investigated and reduced to the atomic level. The present study significantly refines our understanding of the mode of action of NS5B-nucleotide-inhibitors, identifies the key structural elements necessary for their activity, and implements the tools for ranking the potential of additional much needed novel inhibitors of NS5B.