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Sample records for sites fucose residues

  1. Anionic Sites, Fucose Residues and Class I Human Leukocyte Antigen Fate During Interaction of Toxoplasma gondii with Endothelial Cells

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    Stumbo Ana Carolina

    2002-01-01

    Full Text Available Toxoplasma gondii invades and proliferates in human umbilical vein endothelial cells where it resides in a parasitophorous vacuole. In order to analyze which components of the endothelial cell plasma membrane are internalized and become part of the parasitophorous vacuole membrane, the culture of endothelial cells was labeled with cationized ferritin or UEA I lectin or anti Class I human leukocytte antigen (HLA before or after infection with T. gondii. The results showed no cationized ferritin and UEA I lectin in any parasitophorous vacuole membrane, however, the Class I HLA molecule labeling was observed in some endocytic vacuoles containing parasite until 1 h of interaction with T. gondii. After 24 h parasite-host cell interaction, the labeling was absent on the vacuolar membrane, but presents only in small vesicles near parasitophorous vacuole. These results suggest the anionic site and fucose residues are excluded at the time of parasitophorous vacuole formation while Class I HLA molecules are present only on a minority of Toxoplasma-containig vacuoles.

  2. Six independent fucose-binding sites in the crystal structure of Aspergillus oryzae lectin

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    Makyio, Hisayoshi [Structural Biology Research Center, Photon Factory, Institute of Materials Structure Science, High Energy Accelerator Research Organization (KEK), 1-1 Oho, Tsukuba, Ibaraki, 305-0801 (Japan); Shimabukuro, Junpei; Suzuki, Tatsuya [Department of Applied Bioorganic Chemistry, Gifu University, 1-1 Yanagido, Gifu-shi, Gifu 501-1193 (Japan); Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Yoshida Ushinomiya-cho, Sakyo-ku, Kyoto 606-8501 (Japan); Imamura, Akihiro; Ishida, Hideharu [Department of Applied Bioorganic Chemistry, Gifu University, 1-1 Yanagido, Gifu-shi, Gifu 501-1193 (Japan); Kiso, Makoto [Department of Applied Bioorganic Chemistry, Gifu University, 1-1 Yanagido, Gifu-shi, Gifu 501-1193 (Japan); Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Yoshida Ushinomiya-cho, Sakyo-ku, Kyoto 606-8501 (Japan); Ando, Hiromune, E-mail: hando@gifu-u.ac.jp [Department of Applied Bioorganic Chemistry, Gifu University, 1-1 Yanagido, Gifu-shi, Gifu 501-1193 (Japan); Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Yoshida Ushinomiya-cho, Sakyo-ku, Kyoto 606-8501 (Japan); Kato, Ryuichi, E-mail: ryuichi.kato@kek.jp [Structural Biology Research Center, Photon Factory, Institute of Materials Structure Science, High Energy Accelerator Research Organization (KEK), 1-1 Oho, Tsukuba, Ibaraki, 305-0801 (Japan)

    2016-08-26

    The crystal structure of AOL (a fucose-specific lectin of Aspergillus oryzae) has been solved by SAD (single-wavelength anomalous diffraction) and MAD (multi-wavelength anomalous diffraction) phasing of seleno-fucosides. The overall structure is a six-bladed β-propeller similar to that of other fucose-specific lectins. The fucose moieties of the seleno-fucosides are located in six fucose-binding sites. Although the Arg and Glu/Gln residues bound to the fucose moiety are common to all fucose-binding sites, the amino-acid residues involved in fucose binding at each site are not identical. The varying peak heights of the seleniums in the electron density map suggest that each fucose-binding site has a different carbohydrate binding affinity. - Highlights: • The six-bladed β-propeller structure of AOL was solved by seleno-sugar phasing. • The mode of fucose binding is essentially conserved at all six binding sites. • The seleno-fucosides exhibit slightly different interactions and electron densities. • These findings suggest that the affinity for fucose is not identical at each site.

  3. Six independent fucose-binding sites in the crystal structure of Aspergillus oryzae lectin

    International Nuclear Information System (INIS)

    Makyio, Hisayoshi; Shimabukuro, Junpei; Suzuki, Tatsuya; Imamura, Akihiro; Ishida, Hideharu; Kiso, Makoto; Ando, Hiromune; Kato, Ryuichi

    2016-01-01

    The crystal structure of AOL (a fucose-specific lectin of Aspergillus oryzae) has been solved by SAD (single-wavelength anomalous diffraction) and MAD (multi-wavelength anomalous diffraction) phasing of seleno-fucosides. The overall structure is a six-bladed β-propeller similar to that of other fucose-specific lectins. The fucose moieties of the seleno-fucosides are located in six fucose-binding sites. Although the Arg and Glu/Gln residues bound to the fucose moiety are common to all fucose-binding sites, the amino-acid residues involved in fucose binding at each site are not identical. The varying peak heights of the seleniums in the electron density map suggest that each fucose-binding site has a different carbohydrate binding affinity. - Highlights: • The six-bladed β-propeller structure of AOL was solved by seleno-sugar phasing. • The mode of fucose binding is essentially conserved at all six binding sites. • The seleno-fucosides exhibit slightly different interactions and electron densities. • These findings suggest that the affinity for fucose is not identical at each site.

  4. Enhancing GDP-fucose production in recombinant Escherichia coli by metabolic pathway engineering.

    Science.gov (United States)

    Zhai, Yafei; Han, Donglei; Pan, Ying; Wang, Shuaishuai; Fang, Junqiang; Wang, Peng; Liu, Xian-wei

    2015-02-01

    Guanosine 5'-diphosphate (GDP)-fucose is the indispensible donor substrate for fucosyltransferase-catalyzed synthesis of fucose-containing biomolecules, which have been found involving in various biological functions. In this work, the salvage pathway for GDP-fucose biosynthesis from Bacterioides fragilis was introduced into Escherichia coli. Besides, the biosynthesis of guanosine 5'-triphosphate (GTP), an essential substrate for GDP-fucose biosynthesis, was enhanced via overexpression of enzymes involved in the salvage pathway of GTP biosynthesis. The production capacities of metabolically engineered strains bearing different combinations of recombinant enzymes were compared. The shake flask fermentation of the strain expressing Fkp, Gpt, Gmk and Ndk obtained the maximum GDP-fucose content of 4.6 ± 0.22 μmol/g (dry cell mass), which is 4.2 fold that of the strain only expressing Fkp. Through fed-batch fermentation, the GDP-fucose content further rose to 6.6 ± 0.14 μmol/g (dry cell mass). In addition to a better productivity than previous fermentation processes based on the de novo pathway for GDP-fucose biosynthesis, the established schemes in this work also have the advantage to be a potential avenue to GDP-fucose analogs encompassing chemical modification on the fucose residue. Copyright © 2014 Elsevier Inc. All rights reserved.

  5. Knowledge-based modeling of a legume lectin and docking of the carbohydrate ligand: the Ulex europaeus lectin I and its interaction with fucose.

    Science.gov (United States)

    Gohier, A; Espinosa, J F; Jimenez-Barbero, J; Carrupt, P A; Pérez, S; Imberty, A

    1996-12-01

    Ulex europaeus isolectin I is specific for fucose-containing oligosaccharide such as H type 2 trisaccharide alpha-L-Fuc (1-->2) beta-D-Gal (1-->4) beta-D-GlcNAc. Several legume lectins have been crystallized and modeled, but no structural data are available concerning such fucose-binding lectin. The three-dimensional structure of Ulex europaeus isolectin I has been constructed using seven legume lectins for which high-resolution crystal structures were available. Some conserved water molecules, as well as the structural cations, were taken into account for building the model. In the predicted binding site, the most probable locations of the secondary hydroxyl groups were determined using the GRID method. Several possible orientations could be determined for a fucose residue. All of the four possible conformations compatible with energy calculations display several hydrogen bonds with Asp-87 and Ser-132 and a stacking interaction with Tyr-220 and Phe-136. In two orientations, the O-3 and O-4 hydroxyl groups of fucose are the most buried ones, whereas two other, the O-2 and O-3 hydroxyl groups are at the bottom of the site. Possible docking modes are also studied by analysis of the hydrophobic and hydrophilic surfaces for both the ligand and the protein. The SCORE method allows for a quantitative evaluation of the complementarity of these surfaces, on the basis of molecular lipophilicity calculations. The predictions presented here are compared with known biochemical data.

  6. The crystal structure of human GDP-L-fucose synthase.

    Science.gov (United States)

    Zhou, Huan; Sun, Lihua; Li, Jian; Xu, Chunyan; Yu, Feng; Liu, Yahui; Ji, Chaoneng; He, Jianhua

    2013-09-01

    Human GDP-l-fucose synthase, also known as FX protein, synthesizes GDP-l-fucose from its substrate GDP-4-keto-6-deoxy-d-mannose. The reaction involves epimerization at both C-3 and C-5 followed by an NADPH-dependent reduction of the carbonyl at C-4. In this paper, the first crystal structure of human FX protein was determined at 2.37 Å resolution. The asymmetric unit of the crystal structure contains four molecules which form two homodimers. Each molecule consists of two domains, a Rossmann-fold NADPH-binding motif and a carboxyl terminal domain. Compared with the Escherichia coli GDP-l-fucose synthase, the overall structures of these two enzymes have four major differences. There are four loops in the structure of human FX protein corresponding to two α-helices and two β-sheets in that of the E. coli enzyme. Besides, there are seven different amino acid residues binding with NAPDH comparing human FX protein with that from E. coli. The structure of human FX reveals the key catalytic residues and could be useful for the design of drugs for the treatment of inflammation, auto-immune diseases, and possibly certain types of cancer.

  7. Contaminated Sites by Residues from Romanian Industry

    International Nuclear Information System (INIS)

    Aurelian, F.; Georgescu, D.; Vacariu, V.; Popescu, M.

    2001-01-01

    Full text: In Romania, the mining industries of ferrous ore, non-ferrous ore, bauxite, coal, chemical fertilisers, the extraction of natural gas and oil, etc. have a considerable weight in economy. The study lets us know that in these industries, in different by-products and waste from the process, some radionuclides come out. The preliminary data, which are determined in these industries, are compared with those data, which are published by the countries from EU. There were determined uranium, radium and thorium from ore and sterile (ferrous, non-ferrous, bauxite, talc, clay, asbestos, barytine, salt, coal and bituminous schist). Also, there have been determined the radium and the uranium content of waters from non-uraniferous mining sites. The level of radioactive noxa has been informative determined in each mine. The level of the concentrations requires a systematic research and an assessment of the risk and the impact against the environment, the workers, and the public. (author)

  8. Fucose-Mediated Transcriptional Activation of the fcs Operon by FcsR in Streptococcus pneumoniae.

    Science.gov (United States)

    Manzoor, Irfan; Shafeeq, Sulman; Afzal, Muhammad; Kuipers, Oscar P

    2015-01-01

    In this study, we explore the impact of fucose on the transcriptome of S. pneumoniae D39. The expression of various genes and operons, including the fucose uptake PTS and utilization operon (fcs operon) was altered in the presence of fucose. By means of quantitative RT-PCR and β-galactosidase analysis, we demonstrate the role of the transcriptional regulator FcsR, present upstream of the fcs operon, as a transcriptional activator of the fcs operon. We also predict a 19-bp putative FcsR regulatory site (5'-ATTTGAACATTATTCAAGT-3') in the promoter region of the fcs operon. The functionality of this predicted FcsR regulatory site was further confirmed by promoter-truncation experiments, where deletion of half of the FscR regulatory site or full deletion led to the abolition of expression of the fcs operon. © 2015 S. Karger AG, Basel.

  9. The Drosophila Neurally Altered Carbohydrate Mutant Has a Defective Golgi GDP-fucose Transporter*

    Science.gov (United States)

    Geisler, Christoph; Kotu, Varshika; Sharrow, Mary; Rendić, Dubravko; Pöltl, Gerald; Tiemeyer, Michael; Wilson, Iain B. H.; Jarvis, Donald L.

    2012-01-01

    Studying genetic disorders in model organisms can provide insights into heritable human diseases. The Drosophila neurally altered carbohydrate (nac) mutant is deficient for neural expression of the HRP epitope, which consists of N-glycans with core α1,3-linked fucose residues. Here, we show that a conserved serine residue in the Golgi GDP-fucose transporter (GFR) is substituted by leucine in nac1 flies, which abolishes GDP-fucose transport in vivo and in vitro. This loss of function is due to a biochemical defect, not to destabilization or mistargeting of the mutant GFR protein. Mass spectrometry and HPLC analysis showed that nac1 mutants lack not only core α1,3-linked, but also core α1,6-linked fucose residues on their N-glycans. Thus, the nac1 Gfr mutation produces a previously unrecognized general defect in N-glycan core fucosylation. Transgenic expression of a wild-type Gfr gene restored the HRP epitope in neural tissues, directly demonstrating that the Gfr mutation is solely responsible for the neural HRP epitope deficiency in the nac1 mutant. These results validate the Drosophila nac1 mutant as a model for the human congenital disorder of glycosylation, CDG-IIc (also known as LAD-II), which is also the result of a GFR deficiency. PMID:22745127

  10. Combining specificity determining and conserved residues improves functional site prediction

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    Gelfand Mikhail S

    2009-06-01

    Full Text Available Abstract Background Predicting the location of functionally important sites from protein sequence and/or structure is a long-standing problem in computational biology. Most current approaches make use of sequence conservation, assuming that amino acid residues conserved within a protein family are most likely to be functionally important. Most often these approaches do not consider many residues that act to define specific sub-functions within a family, or they make no distinction between residues important for function and those more relevant for maintaining structure (e.g. in the hydrophobic core. Many protein families bind and/or act on a variety of ligands, meaning that conserved residues often only bind a common ligand sub-structure or perform general catalytic activities. Results Here we present a novel method for functional site prediction based on identification of conserved positions, as well as those responsible for determining ligand specificity. We define Specificity-Determining Positions (SDPs, as those occupied by conserved residues within sub-groups of proteins in a family having a common specificity, but differ between groups, and are thus likely to account for specific recognition events. We benchmark the approach on enzyme families of known 3D structure with bound substrates, and find that in nearly all families residues predicted by SDPsite are in contact with the bound substrate, and that the addition of SDPs significantly improves functional site prediction accuracy. We apply SDPsite to various families of proteins containing known three-dimensional structures, but lacking clear functional annotations, and discusse several illustrative examples. Conclusion The results suggest a better means to predict functional details for the thousands of protein structures determined prior to a clear understanding of molecular function.

  11. Inventory of sites used to develop residual radioactivity criteria

    International Nuclear Information System (INIS)

    Ronca-Battista, M.; Hardeman, J.C. Jr.

    1986-01-01

    The US Environmental Protection Agency (EPA), in conjunction with the National Conference of Radiation Control Program Directors (CRCPD), has compiled an inventory of never licensed or otherwise poorly documented sites that may be contaminated with radioactive materials. This effort is in support of the EPA's development of radiation protection criteria for residual radioactivity at decommissioned sites. The inventory will help to establish the range of circumstances for which criteria are needed, as well as the suitability of candidate criteria for actual situations. The information will also be used to develop model sites and facilities for analyzing technical and economic feasibility of residual radioactivity criteria and to assess costs and benefits of alternate criteria. Relevant information about each site, such as radionuclides, waste forms, and quantities present will be included in the inventory when such information is available. The CRCPD has requested that each State radiation control agency furnish the information for the inventory. The inventory supplements the relatively extensive documentation of sites regulated by Federal or State agencies with information on old or unlicensed sites, such as old waste storage sites or radium ore processing facilities

  12. Residual herbicide study on selected Hanford Site roadsides

    Energy Technology Data Exchange (ETDEWEB)

    Smith, J.L.; Kemp, C.J.; Sackschewsky, M.R.

    1993-08-01

    Westinghouse Hanford Company routinely treats roadsides with herbicides to control undesirable plant growth. An experiment was conducted to test perennial grass germination in soils adjacent to roadways of the Hanford Site. The primary variable was the distance from the roadside. A simple germination test was executed in a controlled-environment chamber to determine the residual effects of these applications. As expected, the greatest herbicide activity was found directly adjacent to the roadway, approximately 0 to 20 ft (0 to 6.3 m) from the roadway.

  13. Characterization of Active Site Residues of Nitroalkane Oxidase†

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    Valley, Michael P.; Fenny, Nana S.; Ali, Shah R.; Fitzpatrick, Paul F.

    2010-01-01

    The flavoenzyme nitroalkane oxidase catalyzes the oxidation of primary and secondary nitrolkanes to the corresponding aldehydes and ketones plus nitrite. The structure of the enzyme shows that Serl71 forms a hydrogen bond to the flavin N5, suggesting that it plays a role in catalysis. Cys397 and Tyr398 were previously identified by chemical modification as potential active site residues. To more directly probe the roles of these residues, the S171A, S171V, S171T, C397S, and Y398F enzymes have been characterized with nitroethane as substrate. The C397S and Y398 enzymes were less stable than the wild-type enzyme, and the C397S enzyme routinely contained a substoichiometric amount of FAD. Analysis of the steady-state kinetic parameters for the mutant enzymes, including deuterium isotope effects, establishes that all of the mutations result in decreases in the rate constants for removal of the substrate proton by ~5-fold and decreases in the rate constant for product release of ~2-fold. Only the S171V and S171T mutations alter the rate constant for flavin oxidation. These results establish that these residues are not involved in catalysis, but rather are required for maintaining the protein structure. PMID:20056514

  14. Characterization of active site residues of nitroalkane oxidase.

    Science.gov (United States)

    Valley, Michael P; Fenny, Nana S; Ali, Shah R; Fitzpatrick, Paul F

    2010-06-01

    The flavoenzyme nitroalkane oxidase catalyzes the oxidation of primary and secondary nitroalkanes to the corresponding aldehydes and ketones plus nitrite. The structure of the enzyme shows that Ser171 forms a hydrogen bond to the flavin N5, suggesting that it plays a role in catalysis. Cys397 and Tyr398 were previously identified by chemical modification as potential active site residues. To more directly probe the roles of these residues, the S171A, S171V, S171T, C397S, and Y398F enzymes have been characterized with nitroethane as substrate. The C397S and Y398 enzymes were less stable than the wild-type enzyme, and the C397S enzyme routinely contained a substoichiometric amount of FAD. Analysis of the steady-state kinetic parameters for the mutant enzymes, including deuterium isotope effects, establishes that all of the mutations result in decreases in the rate constants for removal of the substrate proton by approximately 5-fold and decreases in the rate constant for product release of approximately 2-fold. Only the S171V and S171T mutations alter the rate constant for flavin oxidation. These results establish that these residues are not involved in catalysis, but rather are required for maintaining the protein structure. 2009 Elsevier Inc. All rights reserved.

  15. Genetic metabolic complementation establishes a requirement for GDP-fucose in Leishmania.

    Science.gov (United States)

    Guo, Hongjie; Novozhilova, Natalia M; Bandini, Giulia; Turco, Salvatore J; Ferguson, Michael A J; Beverley, Stephen M

    2017-06-23

    To survive in its sand fly vector, the trypanosomatid protozoan parasite Leishmania first attaches to the midgut to avoid excretion, but eventually it must detach for transmission by the next bite. In Leishmania major strain Friedlin, this is controlled by modifications of the stage-specific adhesin lipophosphoglycan (LPG). During differentiation to infective metacyclics, d-arabinopyranose (d-Ara p ) caps the LPG side-chain galactose residues, blocking interaction with the midgut lectin PpGalec, thereby leading to parasite detachment and transmission. Previously, we characterized two closely related L. major genes ( FKP40 and AFKP80 ) encoding bifunctional proteins with kinase/pyrophosphorylase activities required for salvage and conversion of l-fucose and/or d-Ara p into the nucleotide-sugar substrates required by glycosyltransferases. Whereas only AFKP80 yielded GDP-d-Ara p from exogenous d-Ara p , both proteins were able to salvage l-fucose to GDP-fucose. We now show that Δ afkp80 - null mutants ablated d-Ara p modifications of LPG as predicted, whereas Δ fkp40 - null mutants resembled wild type (WT). Fucoconjugates had not been reported previously in L. major , but unexpectedly, we were unable to generate fkp40 - / afkp80 - double mutants, unless one of the A/FKPs was expressed ectopically. To test whether GDP-fucose itself was essential for Leishmania viability, we employed "genetic metabolite complementation." First, the trypanosome de novo pathway enzymes GDP-mannose dehydratase (GMD) and GDP-fucose synthetase (GMER) were expressed ectopically; from these cells, the Δ fkp40 - /Δ afkp80 - double mutant was now readily obtained. As expected, the Δ fkp40 - /Δ afkp80 - /+ TbGMD-GMER line lacked the capacity to generate GDP-Ara p , while synthesizing abundant GDP-fucose. These results establish a requirement for GDP-fucose for L. major viability and predict the existence of an essential fucoconjugate(s). © 2017 by The American Society for Biochemistry and

  16. A tool for calculating binding-site residues on proteins from PDB structures

    Directory of Open Access Journals (Sweden)

    Hu Jing

    2009-08-01

    Full Text Available Abstract Background In the research on protein functional sites, researchers often need to identify binding-site residues on a protein. A commonly used strategy is to find a complex structure from the Protein Data Bank (PDB that consists of the protein of interest and its interacting partner(s and calculate binding-site residues based on the complex structure. However, since a protein may participate in multiple interactions, the binding-site residues calculated based on one complex structure usually do not reveal all binding sites on a protein. Thus, this requires researchers to find all PDB complexes that contain the protein of interest and combine the binding-site information gleaned from them. This process is very time-consuming. Especially, combing binding-site information obtained from different PDB structures requires tedious work to align protein sequences. The process becomes overwhelmingly difficult when researchers have a large set of proteins to analyze, which is usually the case in practice. Results In this study, we have developed a tool for calculating binding-site residues on proteins, TCBRP http://yanbioinformatics.cs.usu.edu:8080/ppbindingsubmit. For an input protein, TCBRP can quickly find all binding-site residues on the protein by automatically combining the information obtained from all PDB structures that consist of the protein of interest. Additionally, TCBRP presents the binding-site residues in different categories according to the interaction type. TCBRP also allows researchers to set the definition of binding-site residues. Conclusion The developed tool is very useful for the research on protein binding site analysis and prediction.

  17. Rescue of Notch signaling in cells incapable of GDP-L-fucose synthesis by gap junction transfer of GDP-L-fucose in Drosophila.

    Science.gov (United States)

    Ayukawa, Tomonori; Matsumoto, Kenjiroo; Ishikawa, Hiroyuki O; Ishio, Akira; Yamakawa, Tomoko; Aoyama, Naoki; Suzuki, Takuya; Matsuno, Kenji

    2012-09-18

    Notch (N) is a transmembrane receptor that mediates cell-cell interactions to determine many cell-fate decisions. N contains EGF-like repeats, many of which have an O-fucose glycan modification that regulates N-ligand binding. This modification requires GDP-L-fucose as a donor of fucose. The GDP-L-fucose biosynthetic pathways are well understood, including the de novo pathway, which depends on GDP-mannose 4,6 dehydratase (Gmd) and GDP-4-keto-6-deoxy-D-mannose 3,5-epimerase/4-reductase (Gmer). However, the potential for intercellularly supplied GDP-L-fucose and the molecular basis of such transportation have not been explored in depth. To address these points, we studied the genetic effects of mutating Gmd and Gmer on fucose modifications in Drosophila. We found that these mutants functioned cell-nonautonomously, and that GDP-L-fucose was supplied intercellularly through gap junctions composed of Innexin-2. GDP-L-fucose was not supplied through body fluids from different isolated organs, indicating that the intercellular distribution of GDP-L-fucose is restricted within a given organ. Moreover, the gap junction-mediated supply of GDP-L-fucose was sufficient to support the fucosylation of N-glycans and the O-fucosylation of the N EGF-like repeats. Our results indicate that intercellular delivery is a metabolic pathway for nucleotide sugars in live animals under certain circumstances.

  18. Prediction of Active Site and Distal Residues in E. coli DNA Polymerase III alpha Polymerase Activity.

    Science.gov (United States)

    Parasuram, Ramya; Coulther, Timothy A; Hollander, Judith M; Keston-Smith, Elise; Ondrechen, Mary Jo; Beuning, Penny J

    2018-02-20

    The process of DNA replication is carried out with high efficiency and accuracy by DNA polymerases. The replicative polymerase in E. coli is DNA Pol III, which is a complex of 10 different subunits that coordinates simultaneous replication on the leading and lagging strands. The 1160-residue Pol III alpha subunit is responsible for the polymerase activity and copies DNA accurately, making one error per 10 5 nucleotide incorporations. The goal of this research is to determine the residues that contribute to the activity of the polymerase subunit. Homology modeling and the computational methods of THEMATICS and POOL were used to predict functionally important amino acid residues through their computed chemical properties. Site-directed mutagenesis and biochemical assays were used to validate these predictions. Primer extension, steady-state single-nucleotide incorporation kinetics, and thermal denaturation assays were performed to understand the contribution of these residues to the function of the polymerase. This work shows that the top 15 residues predicted by POOL, a set that includes the three previously known catalytic aspartate residues, seven remote residues, plus five previously unexplored first-layer residues, are important for function. Six previously unidentified residues, R362, D405, K553, Y686, E688, and H760, are each essential to Pol III activity; three additional residues, Y340, R390, and K758, play important roles in activity.

  19. Fucose-Mediated Transcriptional Activation of the fcs Operon by FcsR in Streptococcus pneumoniae

    NARCIS (Netherlands)

    Manzoor, Irfan; Shafeeq, Sulman; Afzal, Muhammad; Kuipers, Oscar P

    2015-01-01

    In this study, we explore the impact of fucose on the transcriptome of S. pneumoniae D39. The expression of various genes and operons, including the fucose uptake PTS and utilization operon (fcs operon) was altered in the presence of fucose. By means of quantitative RT-PCR and β-galactosidase

  20. Assessment of residual DDE at four remediated Hanford waste sites, Richland, Washington

    International Nuclear Information System (INIS)

    Linville, J.K.

    1999-01-01

    The objectives of this study were to determine the extent and distribution of residual DDE, a metabolite of dichlorodiphenyltrichloroethane (DDT), across the four waste sites by sampling ground-dwelling insects and bird eggs, evaluating the use of insects for monitoring contamination pathways, and determining the species of passerine birds present and the number of nesting pairs utilizing the waste sites

  1. Derivation of uranium residual radioactive material guidelines for the former Alba Craft Laboratory site, Oxford, Ohio

    International Nuclear Information System (INIS)

    Nimmagadda, M.; Faillace, E.; Yu, C.

    1994-01-01

    Residual radioactive material guidelines for uranium were derived for the former Alba Craft Laboratory site in Oxford, Ohio. This site has been identified for remedial action under the Formerly Utilized Sites Remedial Action Program (FUSRAP) of the US Department of Energy (DOE). Single nuclide and total uranium guidelines were derived on the basis of the requirement that the 50-year committed effective dose equivalent to a hypothetical individual who lives or works in the immediate vicinity of the former Alba Craft Laboratory site should not exceed a dose of 30 mrem/yr following remedial action for the current use and likely future use scenarios or a dose of 100 mrem/yr for less likely future use scenarios (Yu et al. 1993). The DOE residual radioactive material guideline computer code, RESRAD, which implements the methodology described in the DOE manual for implementing residual radioactive material guidelines, was used in this evaluation

  2. Derivation of uranium residual radioactive material guidelines for the Ventron site

    International Nuclear Information System (INIS)

    Loureiro, C.; Yu, C.; Jones, L.

    1992-03-01

    Residual radioactive material guidelines for uranium were derived for the Ventron site in Beverly, Massachusetts. This site has been identified for remedial action under the Formerly Utilized Sites Remedial Action Program of the US Department of Energy (DOE). The derivations for the single radionuclides and the total uranium guidelines were based on the requirement that the 50-year committed effective dose equivalent to a hypothetical individual who lives or works in the immediate vicinity of the Ventron site should not exceed a dose of 100 mrem/yr following remedial action. The DOE residual radioactive material guideline computer code, RESRAD, which implements the methodology described in the DOE manual for implementing residual radioactive material guidelines, was used in this evaluation

  3. Depleted uranium residual radiological risk assessment for Kosovo sites

    International Nuclear Information System (INIS)

    Durante, Marco; Pugliese, Mariagabriella

    2003-01-01

    During the recent conflict in Yugoslavia, depleted uranium rounds were employed and were left in the battlefield. Health concern is related to the risk arising from contamination of areas in Kosovo with depleted uranium penetrators and dust. Although chemical toxicity is the most significant health risk related to uranium, radiation exposure has been allegedly related to cancers among veterans of the Balkan conflict. Uranium munitions are considered to be a source of radiological contamination of the environment. Based on measurements and estimates from the recent Balkan Task Force UNEP mission in Kosovo, we have estimated effective doses to resident populations using a well-established food-web mathematical model (RESRAD code). The UNEP mission did not find any evidence of widespread contamination in Kosovo. Rather than the actual measurements, we elected to use a desk assessment scenario (Reference Case) proposed by the UNEP group as the source term for computer simulations. Specific applications to two Kosovo sites (Planeja village and Vranovac hill) are described. Results of the simulations suggest that radiation doses from water-independent pathways are negligible (annual doses below 30 μSv). A small radiological risk is expected from contamination of the groundwater in conditions of effective leaching and low distribution coefficient of uranium metal. Under the assumptions of the Reference Case, significant radiological doses (>1 mSv/year) might be achieved after many years from the conflict through water-dependent pathways. Even in this worst-case scenario, DU radiological risk would be far overshadowed by its chemical toxicity

  4. Summary of the engineering assessment of radioactive sands and residues, Lowman Site, Lowman, Idaho

    International Nuclear Information System (INIS)

    1981-09-01

    Ford, Bacon and Davis Utah Inc. has reevaluated the Lowman site in order to revise the December 1977 engineering assessment of the problems resulting from the existence of radioactive sands and residues at Lowman, Idaho. This engineering assessment has included the preparation of topographic maps, the performance of core drillings and radiometric measurements sufficient to determine areas and volumes of radioactive sands and residues and radiation exposures of individuals and nearby populations, the investigations of site hydrology and meteorology, and the evaluation and costing of alternative corrective actions. Radon gas released from the 191,000 tons of radioactive sands, residues, and contaminated soils at the Lowman site constitutes the most significant environmental impact, although windblown radioactive sands and external gamma radiation also are factors

  5. Engineering assessment of radioactive sands and residues, Lowman Site, Lowman, Idaho

    International Nuclear Information System (INIS)

    1981-09-01

    Ford, Bacon and Davis Utah Inc. has reevaluated the Lowman site in order to revise the December 1977 engineering assessment of the problems resulting from the existence of radioactive sands and residues at Lowman, Idaho. This engineering assessment has included the preparation of topographic maps, the performance of core drillings and radiometric measurements sufficient to determine areas and volumes of radioactive sands and residues and radiation exposure of individuals and nearby populations, and investigations of site hydrology and meteorology, and the evaluation and costing of alternative corrective actions. Radon gas released from the 191,000 tons of radioactive sands, residues, and contaminated soils at the Lowman site constitutes the most significant environmental impact, although windblown radioactive sands and external gamma radiation also are factors

  6. Essential histidyl residues at the active site(s) of sucrose-phosphate synthase from Prosopis juliflora.

    Science.gov (United States)

    Sinha, A K; Pathre, U V; Sane, P V

    1998-11-10

    Chemical modification of sucrose-phosphate synthase (EC 2.4.1.14) from Prosopis juliflora by diethyl pyrocarbonate (DEP) and photo-oxidation in the presence of rose bengal (RB) which modify the histidyl residues of the protein resulted in the inactivation of the enzyme activity. This inactivation was dependent on the concentration of the modifying reagent and the time of incubation and followed pseudo-first order kinetics. For both the reagents, the inactivation was maximum at pH 7.5, which is consistent with the involvement and presence of histidine residues at the active site of the enzyme. Substrates, UDPG and F6P protected the enzyme against the inactivation by the modifying reagents suggesting that the histidine residues may be involved in the binding of these substrates and are essential for the catalytic activity. Specificity of DEP was indicated by an increase in absorbance at 240 nm along with concomitant inactivation of the enzyme and reactivation of the modified enzyme by hydroxylamine. These results strongly suggest the presence of histidine residue(s) at or near the active site of the enzyme.

  7. Phase II, Title I engineering assessment of radioactive sands and residues, Lowman Site, Lowman Idaho

    International Nuclear Information System (INIS)

    1977-12-01

    An engineering assessment was performed of the problems resulting from the existence of radioactive uranium sand residues at the Lowman, Idaho, site. Services normally include the preparation of topographic maps, the performance of core drillings and radiometric measurements sufficient to determine areas and volumes of tailings and other radium-contaminated materials, the evaluation of resulting investigations of site hydrology and meteorology, and the evaluation and costing of alternative corrective actions. Radon gas release from the 90,000 tons of sand residues at the Lowman site constitutes the most significant environmental impact, although external gamma radiation is also a factor. The two alternative actions presented are dike construction, fencing, and maintenance (Option I); and consolidation of the piles, addition of a 2-ft-thick stabilization cover, and on-site cleanup (Option II). Both options include remedial action at off-site structures. Cost estimates for the two options are $393,000 and $590,000

  8. Phase II, Title I engineering assessment of radioactive sands and residues, Lowman Site, Lowman Idaho

    Energy Technology Data Exchange (ETDEWEB)

    1977-12-01

    An engineering assessment was performed of the problems resulting from the existence of radioactive uranium sand residues at the Lowman, Idaho, site. Services normally include the preparation of topographic maps, the performance of core drillings and radiometric measurements sufficient to determine areas and volumes of tailings and other radium-contaminated materials, the evaluation of resulting investigations of site hydrology and meteorology, and the evaluation and costing of alternative corrective actions. Radon gas release from the 90,000 tons of sand residues at the Lowman site constitutes the most significant environmental impact, although external gamma radiation is also a factor. The two alternative actions presented are dike construction, fencing, and maintenance (Option I); and consolidation of the piles, addition of a 2-ft-thick stabilization cover, and on-site cleanup (Option II). Both options include remedial action at off-site structures. Cost estimates for the two options are $393,000 and $590,000.

  9. Methodology for determining acceptable residual radioactive contamination levels at decommissioned nuclear facilities/sites

    International Nuclear Information System (INIS)

    Watson, E.C.; Kennedy, W.E. Jr.; Hoenes, G.R.; Waite, D.A.

    1979-01-01

    The ultimate disposition of decommissioned nuclear facilities and their surrrounding sites depends upon the degree and type of residual contamination. Examination of existing guidelines and regulations has led to the conclusion that there is a need for a general method to derive residual radioactive contamination levels that are acceptable for public use of any decommissioned nuclear facility or site. This paper describes a methodology for determining acceptable residual radioactive contamination levels based on the concept of limiting the annual dose to members of the public. It is not the purpose of this paper to recommend or even propose dose limits for the exposure of the public to residual radioactive contamination left at decommissioned nuclear facilities or sites. Unrestricted release of facilities and/or land is based on the premise that the potential annual dose to any member of the public using this property from all possible exposure pathways will not exceed appropriate limits as may be defined by Federal regulatory agencies. For decommissioned land areas, consideration should be given to people living directly on previously contaminated areas, growing crops, grazing food animals and using well water. Mixtures of radionuclides in the residual contamination representative of fuel reprocessing plants, light water reactors and their respective sites are presented. These mixtures are then used to demonstrate the methodology. Example acceptable residual radioactive contamination levels, based on an assumed maximum annual dose of one millirem, are calculated for several selected times following shutdown of a facility. It is concluded that the methodology presented in this paper results in defensible acceptable residual contamination levels that are directly relatable to risk assessment with the proviso that an acceptable limit to the maximum annual dose will be established. (author)

  10. Derivation of residual radioactive material guidelines for the Laboratory for Energy-Related Health Research site

    International Nuclear Information System (INIS)

    Chapman, T.E.

    1993-11-01

    Residual radioactive material guidelines were derived for the Laboratory for Energy-Related Health Research (LEHR) Environmental Restoration (ER) site in Davis, California. The guideline derivation was based on a dose limit of 100 mrem/yr. The US Department of Energy (DOE) residual radioactive material guideline computer code was used in this evaluation. This code implements the methodology described in the DOE manual for implementing residual radioactive material guidelines. Three potential site utilization scenarios were considered with the assumption that following ER action, the site will be used without radiological restrictions. The defined scenarios vary with regard to use of the site, time spent at the site, and sources of food consumed. The results of the evaluation indicate that the basic dose limit of 100 mrem/yr will not be exceeded, provided that the soil concentrations of these radionuclides at the LEHR site do not exceed the scenario-specific values calculated by this study. Except for the extent of the contaminated zone (which is very conservative), assumptions used are as site-specific as possible, given available information. The derived guidelines are single- radionuclide guidelines and are linearly proportional to the dose limit used in the calculations. In setting the actual residual soil contamination guides for the LEHR site, DOE will apply the as low as reasonably achievable policy to the decision-making process, along with other factors such as whether a particular scenario is reasonable and appropriate, as well as using site-specific inputs to computer models based on data not yet fully determined

  11. Status policy and criteria for the remediation of sites with radioactive residues in China

    International Nuclear Information System (INIS)

    Yamin, Z.

    1999-01-01

    This paper introduces the status policy and criteria for remediation of sites with radioactive residues in China. We deal with the sites in difference patterns according to their circumstances. For the sites related with the decommissioning of the nuclear fuel cycle facilities, the programs for environmental restoration must be reviewed and approved by the State Environmental Protection Administration (SEPA). And the radioactive waste resulting from these facilities should be collected and eventually disposed of at the regional disposal repositories built by the country. For the sites related with use of radionuclides in medicine, research and industry, as well as the small scale extraction and processing of materials containing natural radionuclides, the provincial environmental authority is responsible for approving the restoration projects. These radioactive wastes should finally be sent to the special radioactive waste repositories built by the provincial environmental authorities. So far 21 waste repositories have been built in some provinces. More then 10 sites with radioactive residues have been restored. The standards for general public dose limit and soil residual radionuclides content have been established. (author)

  12. Derivation of uranium residual radioactive material guidelines for the Shpack site

    International Nuclear Information System (INIS)

    Cheng, J.J.; Yu, C.; Monette, F.; Jones, L.

    1991-08-01

    Residual radioactive material guidelines for uranium were derived for the Shpack site in Norton, Massachusetts. This site has been identified for remedial action under the Formerly Utilized Sites Remedial Action Program (FUSRAP) of the US Department of Energy (DOE). The uranium guidelines were derived on the basis of the requirement that the 50-year committed effective dose equivalent to a hypothetical individual who lives or works in the immediate vicinity of the Shpack site should not exceed a dose of 100 mrem/yr following decontamination. The DOE residual radioactive material guideline computer code, RESRAD, which implements the methodology described in the DOE manual for implementing residual radioactive material guidelines, was used in this evaluation. Three potential scenarios were considered for the site; the scenarios vary with regard to time spent at the site, sources of water used, and sources of food consumed. The results of the evaluation indicate that the basic dose limit of 100 mrem/yr will not be exceeded for uranium (including uranium-234, uranium-235, and uranium-238) within 1000 years, provided that the soil concentration of combined uranium (uranium-234 and uranium-238) at the Shpack site does not exceed the following levels: 2500 pCi/g for Scenario A (recreationist: the expected scenario); 1100 pCi/g for Scenario B (industrial worker: a plausible scenario); and 53 pCi/g for Scenario C (resident farmer using a well water as the only water source: a possible but unlikely scenario). The uranium guidelines derived in this report apply to the combined activity concentration of uranium-234 and uranium-238 and were calculated on the basis of a dose of 100 mrem/yr. In setting the actual uranium guidelines for the Shpack site, DOE will apply the as low as reasonably achievable (ALARA) policy to the decision-making process, along with other factors, such as whether a particular scenario is reasonable and appropriate. 8 refs., 2 figs., 8 tabs

  13. Derivation of cesium-137 residual radioactive material guidelines for the Peek Street site, Schenectady, New York

    International Nuclear Information System (INIS)

    Jones, L.; Nimmagadda, M.; Yu, C.

    1992-01-01

    Residual radioactive material guidelines for cesium-137 were derived for the Peek rk. The derivation was based on the requirement that the Street site in Schenectady, New York. The derivation was based on the requirement that the 50-year committed effective dose equivalent to a hypothetical individual who lives or works in the immediate vicinity of the Peek Street site should not exceed a dose of 100 mrem/yr following remedial action. The US Department of Energy (DOE) residual radioactive material guideline computer code, RESRAD was used in this evaluation. Three potential scenarios were considered for the site on the assumption that for a period of 1,000 years following remedial action, the site wig be utilized without radiological restrictions. The scenarios vary with regard to use of the site, time spent at the site, and sources of food consumed. Results indicate that the basic dose limit of 100 mrem/yr will not be exceeded for cesium-137 within 1,000 years, provided that the soil concentration of cesium-137 at the Peek Street site does not exceed the following levels: 98 pCi/g for Scenario A (industrial worker: the expected scenario), 240 pCi/g for Scenario B (recreationist: a plausible scenario), and 34 pCi/g for Scenario C (resident farmer ingesting food produced in the decontaminated area: a plausible scenario)

  14. The Arabidopsis Golgi-localized GDP-L-fucose transporter is required for plant development

    DEFF Research Database (Denmark)

    Rautengarten, Carsten; Ebert, Berit; Liu, Lifeng

    2016-01-01

    assays, we show that GFT preferentially transports GDP-L-fucose over other nucleotide sugars in vitro, while GFT1-silenced plants are almost devoid of L-fucose in cell wall-derived xyloglucan and rhamnogalacturonan II. Furthermore, these lines display reduced L-fucose content in N-glycan structures......Nucleotide sugar transport across Golgi membranes is essential for the luminal biosynthesis of glycan structures. Here we identify GDP-fucose transporter 1 (GFT1), an Arabidopsis nucleotide sugar transporter that translocates GDP-L-fucose into the Golgi lumen. Using proteo-liposome-based transport...... accompanied by severe developmental growth defects. We conclude that GFT1 is the major nucleotide sugar transporter for import of GDP-L-fucose into the Golgi and is required for proper plant growth and development....

  15. The Arabidopsis Golgi-localized GDP-L-fucose transporter is required for plant development.

    Science.gov (United States)

    Rautengarten, Carsten; Ebert, Berit; Liu, Lifeng; Stonebloom, Solomon; Smith-Moritz, Andreia M; Pauly, Markus; Orellana, Ariel; Scheller, Henrik Vibe; Heazlewood, Joshua L

    2016-07-06

    Nucleotide sugar transport across Golgi membranes is essential for the luminal biosynthesis of glycan structures. Here we identify GDP-fucose transporter 1 (GFT1), an Arabidopsis nucleotide sugar transporter that translocates GDP-L-fucose into the Golgi lumen. Using proteo-liposome-based transport assays, we show that GFT preferentially transports GDP-L-fucose over other nucleotide sugars in vitro, while GFT1-silenced plants are almost devoid of L-fucose in cell wall-derived xyloglucan and rhamnogalacturonan II. Furthermore, these lines display reduced L-fucose content in N-glycan structures accompanied by severe developmental growth defects. We conclude that GFT1 is the major nucleotide sugar transporter for import of GDP-L-fucose into the Golgi and is required for proper plant growth and development.

  16. Engineering evaluation of alternatives for the disposition of Niagara Falls Storage Site, its residues and wastes

    International Nuclear Information System (INIS)

    1984-01-01

    The final disposition scenarios selected by DOE for assessment in this document are consistent with those stated in the Notice of Intent to prepare an Environmental Impact Statement (EIS) for the Niagara Falls Storage Site (NFSS) (DOE, 1983d) and the modifications to the alternatives resulting from the public scoping process. The scenarios are: take no action beyond interim remedial measures other than maintenance and surveillance of the NFSS; retain and manage the NFSS as a long-term waste management facility for the wastes and residues on the site; decontaminate, certify, and release the NFSS for other use, with long-term management of the wastes and residues at other DOE sites; and partially decontaminate the NFSS by removal and transport off site of only the more radioactive residues, and upgrade containment of the remaining wastes and residues on site. The objective of this document is to present to DOE the conceptual engineering, occupational radiation exposure, construction schedule, maintenance and surveillance requirements, and cost information relevant to design and implementation of each of the four scenarios. The specific alternatives within each scenario used as the basis for discussion in this document were evaluated on the bases of engineering considerations, technical feasibility, and regulatory requirements. Selected alternatives determined to be acceptable for each of the four final disposition scenarios for the NFSS were approved by DOE to be assessed and costed in this document. These alternatives are also the subject of the EIS for the NFSS currently being prepared by Argonne National Laboratory (ANL). 40 figures, 38 tables

  17. Enhanced production of GDP-L-fucose by overexpression of NADPH regenerator in recombinant Escherichia coli.

    Science.gov (United States)

    Lee, Won-Heong; Chin, Young-Wook; Han, Nam Soo; Kim, Myoung-Dong; Seo, Jin-Ho

    2011-08-01

    Biosynthesis of guanosine 5'-diphosphate-L-fucose (GDP-L-fucose) requires NADPH as a reducing cofactor. In this study, endogenous NADPH regenerating enzymes such as glucose-6-phosphate dehydrogenase (G6PDH), isocitrate dehydrogenase (Icd), and NADP(+)-dependent malate dehydrogenase (MaeB) were overexpressed to increase GDP-L-fucose production in recombinant Escherichia coli. The effects of overexpression of each NADPH regenerating enzyme on GDP-L-fucose production were investigated in a series of batch and fed-batch fermentations. Batch fermentations showed that overexpression of G6PDH was the most effective for GDP-L-fucose production. However, GDP-L-fucose production was not enhanced by overexpression of G6PDH in the glucose-limited fed-batch fermentation. Hence, a glucose feeding strategy was optimized to enhance GDP-L-fucose production. Fed-batch fermentation with a pH-stat feeding mode for sufficient supply of glucose significantly enhanced GDP-L-fucose production compared with glucose-limited fed-batch fermentation. A maximum GDP-L-fucose concentration of 235.2 ± 3.3 mg l(-1), corresponding to a 21% enhancement in the GDP-L-fucose production compared with the control strain overexpressing GDP-L-fucose biosynthetic enzymes only, was achieved in the pH-stat fed-batch fermentation of the recombinant E. coli overexpressing G6PDH. It was concluded that sufficient glucose supply and efficient NADPH regeneration are crucial for NADPH-dependent GDP-L-fucose production in recombinant E. coli.

  18. Glycoproteins of axonal transport: affinity chromatography on fucose-specific lectins

    Energy Technology Data Exchange (ETDEWEB)

    Gustavsson, S.; Ohlson, C.; Karlsson, J.O.

    1982-03-01

    Rapidly transported fucose-labeled glycoproteins from axons of rabbit retinal ganglion cells were solubilized with nonionic detergents. The solubilized components were subjected to affinity chromatography on three different fucose-specific lectins. A recently characterized fucose-specific lectin from Aleuria aurantia bound reversibly approximately 60% of the applied protein-bound radioactivity. The lectins from Lotus tetragonolobus and Ulex europaeus bound are very small proportions of the labeled rapidly transported glycoproteins.

  19. Characterization of active-site residues of the NIa protease from tobacco vein mottling virus.

    Science.gov (United States)

    Hwang, D C; Kim, D H; Lee, J S; Kang, B H; Han, J; Kim, W; Song, B D; Choi, K Y

    2000-10-31

    Nuclear inclusion a (NIa) protease of tobacco vein mottling virus is responsible for the processing of the viral polyprotein into functional proteins. In order to identify the active-site residues of the TVMV NIa protease, the putative active-site residues, His-46, Asp-81 and Cys-151, were mutated individually to generate H46R, H46A, D81E, D81N, C151S, and C151A, and their mutational effects on the proteolytic activities were examined. Proteolytic activity was completely abolished by the mutations of H46R, H46A, D81N, and C151A, suggesting that the three residues are crucial for catalysis. The mutation of D81E decreased kcat marginally by about 4.7-fold and increased Km by about 8-fold, suggesting that the aspartic acid at position 81 is important for substrate binding but can be substituted by glutamate without any significant decrease in catalysis. The replacement of Cys-151 by Ser to mimic the catalytic triad of chymotrypsin-like serine protease resulted in the drastic decrease in kcat by about 1,260-fold. This result might be due to the difference of the active-site geometry between the NIa protease and chymotrypsin. The protease exhibited a bell-shaped pH-dependent profile with a maximum activity approximately at pH 8.3 and with the abrupt changes at the respective pKa values of approximately 6.6 and 9.2, implying the involvement of a histidine residue in catalysis. Taken together, these results demonstrate that the three residues, His-46, Asp-81, and Cys-151, play a crucial role in catalysis of the TVMV NIa protease.

  20. Roles of s3 site residues of nattokinase on its activity and substrate specificity.

    Science.gov (United States)

    Wu, Shuming; Feng, Chi; Zhong, Jin; Huan, Liandong

    2007-09-01

    Nattokinase (Subtilisin NAT, NK) is a bacterial serine protease with high fibrinolytic activity. To probe their roles on protease activity and substrate specificity, three residues of S3 site (Gly(100), Ser(101) and Leu(126)) were mutated by site-directed mutagenesis. Kinetics parameters of 20 mutants were measured using tetrapeptides as substrates, and their fibrinolytic activities were determined by fibrin plate method. Results of mutation analysis showed that Gly(100) and Ser(101) had reverse steric and electrostatic effects. Residues with bulky or positively charged side chains at position 100 decreased the substrate binding and catalytic activity drastically, while residues with the same characters at position 101 could obviously enhance protease and fibrinolytic activity of NK. Mutation of Leu(126) might impair the structure of the active cleft and drastically decreased the activity of NK. Kinetics studies of the mutants showed that S3 residues were crucial to keep protease activity while they moderately affected substrate specificity of NK. The present study provided some original insight into the P3-S3 interaction in NK and other subtilisins, as well as showed successful protein engineering cases to improve NK as a potential therapeutic agent.

  1. Modulation of guanosine nucleotides biosynthetic pathways enhanced GDP-L-fucose production in recombinant Escherichia coli.

    Science.gov (United States)

    Lee, Won-Heong; Shin, So-Yeon; Kim, Myoung-Dong; Han, Nam Soo; Seo, Jin-Ho

    2012-03-01

    Guanosine 5'-triphosphate (GTP) is the key substrate for biosynthesis of guanosine 5'-diphosphate (GDP)-L-fucose. In this study, improvement of GDP-L-fucose production was attempted by manipulating the biosynthetic pathway for guanosine nucleotides in recombinant Escherichia coli-producing GDP-L-fucose. The effects of overexpression of inosine 5'-monophosphate (IMP) dehydrogenase, guanosine 5'-monophosphate (GMP) synthetase (GuaB and GuaA), GMP reductase (GuaC) and guanosine-inosine kinase (Gsk) on GDP-L-fucose production were investigated in a series of fed-batch fermentations. Among the enzymes tested, overexpression of Gsk led to a significant improvement of GDP-L-fucose production. Maximum GDP-L-fucose concentration of 305.5 ± 5.3 mg l(-1) was obtained in the pH-stat fed-batch fermentation of recombinant E. coli-overexpressing Gsk, which corresponds to a 58% enhancement in the GDP-L-fucose production compared with the control strain overexpressing GDP-L-fucose biosynthetic enzymes. Such an enhancement of GDP-L-fucose production could be due to the increase in the intracellular level of GMP.

  2. Residual radioactivity in the soil of the Semipalatinsk Nuclear Test Site in the former USSR

    International Nuclear Information System (INIS)

    Yamamoto, Masayoshi; Tsukatani, Tsuneo; Katayama, Yukio

    1996-01-01

    This paper deals with our efforts to survey residual readioactivity in the soil sampled at teh Semipalatinsk Nuclear Test Site and at off-site areas in Kazakhstan. The soil sampled at the hypocenter where the first Soviet nuclear explosion was carried out on 29 August 1949, and at the bank of the crater called open-quotes Bolapan,close quotes which was formed by an underground nuclear detonation on 15 January 1965 along the Shagan River. As a comparison, other soil was also sampled in the cities of Kurchatov and Almaty. These data have allowed a preliminary evaluation of the contemporary radioactive contamination of the land in and around the test site. At the first nuclear explosion site and at Bolapan, higher than background levels of 239,240 Pu with weapons-grade plutonium were detected together with fission and activation products such as 137 Cs, 60 Co, 152 Eu, and 154 Eu. 20 refs., 3 figs., 5 tabs

  3. Residual radioactivity in the soil of the Semipalatinsk Nuclear Test Site in the former USSR.

    Science.gov (United States)

    Yamamoto, M; Tsukatani, T; Katayama, Y

    1996-08-01

    This paper deals with our efforts to survey residual radioactivity in the soil sampled at the Semipalatinsk Nuclear Test Site and at off-site areas in Kazakhstan. The soil was sampled at the hypocenter where the first Soviet nuclear explosion was carried out on 29 August 1949, and at the bank of the crater called "Bolapan," which was formed by an underground nuclear detonation on 15 January 1965 along the Shagan River. As a comparison, other soil was also sampled in the cities of Kurchatov and Almaty. These data have allowed a preliminary evaluation of the contemporary radioactive contamination of the land in and around the test site. At the first nuclear explosion site and at Bolapan, higher than background levels of 239,240Pu with weapons-grade plutonium were detected together with fission and activation products such as 137Cs, 60Co, 152Eu, and 154Eu.

  4. Structures of NodZ α1,6-fucosyltransferase in complex with GDP and GDP-fucose

    Energy Technology Data Exchange (ETDEWEB)

    Brzezinski, Krzysztof [Argonne National Laboratory, Argonne, IL 60439 (United States); Polish Academy of Sciences, 61-704 Poznan (Poland); Dauter, Zbigniew [Argonne National Laboratory, Argonne, IL 60439 (United States); Jaskolski, Mariusz, E-mail: mariuszj@amu.edu.pl [Polish Academy of Sciences, 61-704 Poznan (Poland); A. Mickiewicz University, 60-780 Poznan (Poland); Argonne National Laboratory, Argonne, IL 60439 (United States)

    2012-02-01

    Crystal structures of the bacterial α1,6-fucosyltransferase NodZ in complex with GDP and GDP-fucose are presented. Rhizobial NodZ α1,6-fucosyltransferase (α1,6-FucT) catalyzes the transfer of the fucose (Fuc) moiety from guanosine 5′-diphosphate-β-l-fucose to the reducing end of the chitin oligosaccharide core during Nod-factor (NF) biosynthesis. NF is a key signalling molecule required for successful symbiosis with a legume host for atmospheric nitrogen fixation. To date, only two α1,6-FucT structures have been determined, both without any donor or acceptor molecule that could highlight the structural background of the catalytic mechanism. Here, the first crystal structures of α1,6-FucT in complex with its substrate GDP-Fuc and with GDP, which is a byproduct of the enzymatic reaction, are presented. The crystal of the complex with GDP-Fuc was obtained through soaking of native NodZ crystals with the ligand and its structure has been determined at 2.35 Å resolution. The fucose residue is exposed to solvent and is disordered. The enzyme–product complex crystal was obtained by cocrystallization with GDP and an acceptor molecule, penta-N-acetyl-l-glucosamine (penta-NAG). The structure has been determined at 1.98 Å resolution, showing that only the GDP molecule is present in the complex. In both structures the ligands are located in a cleft formed between the two domains of NodZ and extend towards the C-terminal domain, but their conformations differ significantly. The structures revealed that residues in three regions of the C-terminal domain, which are conserved among α1,2-, α1,6- and protein O-fucosyltransferases, are involved in interactions with the sugar-donor molecule. There is also an interaction with the side chain of Tyr45 in the N-terminal domain, which is very unusual for a GT-B-type glycosyltransferase. Only minor conformational changes of the protein backbone are observed upon ligand binding. The only exception is a movement of the loop

  5. Contaminant Release from Residual Waste in Single Shell Tanks at the Hanford Site, Washington, USA - 9276

    International Nuclear Information System (INIS)

    Cantrell, Kirk J.; Krupka, Kenneth M.; Deutsch, William J.; Lindberg, Michael J.

    2009-01-01

    Determinations of elemental and solid-phase compositions, and contaminant release studies have been applied in an ongoing study of residual tank wastes (i.e., waste remaining after final retrieval operations) from five of 149 underground single-shell storage tanks (241-C-103, 241-C-106, 241-C-202, 241-C-203, and 241-S-112) at the U.S. Department of Energy's Hanford Site in Washington State. This work is being conducted to support performance assessments that will be required to evaluate long-term health and safety risks associated with tank site closure. The results of studies completed to date show significant variability in the compositions, solid phase properties, and contaminant release characteristics from these residual tank wastes. This variability is the result of differences in waste chemistry/composition of wastes produced from several different spent fuel reprocessing schemes, subsequent waste reprocessing to remove certain target constituents, tank farm operations that concentrated wastes and mixed wastes between tanks, and differences in retrieval processes used to remove the wastes from the tanks. Release models were developed based upon results of chemical characterization of the bulk residual waste, solid-phase characterization (see companion paper 9277 by Krupka et al.), leaching and extraction experiments, and geochemical modeling. In most cases empirical release models were required to describe contaminant release from these wastes. Release of contaminants from residual waste was frequently found to be controlled by the solubility of phases that could not be identified and/or for which thermodynamic data and/or dissolution rates have not been measured. For example, significant fractions of Tc-99, I-129, and Cr appear to be coprecipitated at trace concentrations in metal oxide phases that could not be identified unambiguously. In the case of U release from tank 241-C-103 residual waste, geochemical calculations indicated that leachate

  6. Identification of a highly reactive threonine residue at the active site of γ-glutamyl transpeptidase

    International Nuclear Information System (INIS)

    Stole, E.; Seddon, A.P.; Wellner, D.; Meister, A.

    1990-01-01

    γ-Glutamyl transpeptidase an enzyme of major importance in glutathione metabolism, was inactivated by treating it with L-(αS,5S)-α-amino-3-chloro-4,5-dihydro-5-[3- 14 C]isoxazoleacetic acid. This selective reagent binds stoichiometrically to the enzyme; more than 90% of the label was bound to its light subunit. Enzymatic digestion of the light subunit gave a 14 C-labeled peptide that corresponds to amino acid residues 517-527 of the enzyme and two incomplete digestion products that contain this labeled peptide moiety. The radioactivity associated with this peptide was released with threonine-523 during sequencing by the automated gas-phase Edman method. The light subunit contains 14 other threonine residues and a total of 19 serine residues; these were not labeled. Threonine-523 is situated in the enzyme in an environment that greatly increases its reactivity, indicating that other amino acid residues of the enzyme must also participate in the active-site chemistry of the enzyme

  7. On-site treatment and landfilling of MSWI air pollution control residues

    DEFF Research Database (Denmark)

    Lundtorp, Kasper; Jensen, Dorthe Lærke; Sørensen, Mette Abildgaard

    2003-01-01

    the process, collected through the drainage system, contained large concentrations of salts (Cl: 14–30 g/l, Na: 4–9 g/l, K: 5–11 g/l, Ca: 2–12 g/l) but low concentrations of trace metals (e.g. Pb: 14–100 μg/l, Cd: leaching......Air pollution control (APC) residues from municipal solid waste incineration (MSWI) are difficult to landfill due to substantial leaching of trace metals. An on-site pretreatment prior to landfilling of APC-residues was investigated in terms of bench-scale experiments with a semidry APC...... of the leaching, concentrations of trace metals were reduced by up to four orders of magnitude....

  8. Proposed plan for remedial action at the quarry residuals operable unit of the Weldon Spring Site

    International Nuclear Information System (INIS)

    1998-03-01

    This proposed plan addresses the management of contamination present in various components of the quarry residuals operable unit (QROU) of the Weldon Spring site, which is located in St. Charles County, Missouri. The QROU consists of (1) residual waste at the quarry proper; (2) the Femme Osage Slough, Little Femme Osage Creek, and Femme Osage Creek; and (3) quarry groundwater located primarily north of the slough. Potential impacts to the St. Charles County well field downgradient of the quarry area are also being addressed as part of the evaluations for this operable unit. Remedial activities for the QROU will be conducted by the US Department of Energy (DOE) in accordance with the Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA), as amended. As part of the remedial investigation/feasibility study (RI/FS) process required for the QROU under CERCLA, three major evaluation documents have been prepared to support cleanup decisions for this operable unit. decisions for this operable unit

  9. Derivation of uranium residual radioactive material guidelines for the Elza Gate Site

    International Nuclear Information System (INIS)

    Cheng, J.J.; Yu, C.; Devgun, J.S.

    1991-02-01

    Residual radioactive material guidelines for uranium were derived for a large, homogeneously contaminated area at the Elza Gate Site in Oak Ridge, Tennessee. The derivation of the single-nuclide and total uranium guidelines was based on the requirement that the 50-year committed effective dose equivalent to hypothetical individual who lives or works in the immediate vicinity of the Elza Gate Site should not exceed a dose of 100 mrem/yr following decontamination. The DOE residual radioactive guideline computer code RESRAD was used in this evaluation. Four potential scenarios were considered for the site; the scenarios vary with regard to time spent at the site, sources of water used, and sources of food consumed. The results of the evaluation indicate that the basic dose limit of 100 mrem/yr will not be exceeded for uranium within 1000 years, provided that the soil concentration of uranium at the Elza Gate Site does not exceed the following levels: 1800 pCi/g for Scenario A (industrial worker: the expected scenario); 4000 pCi/g for Scenario B (recreationist: a plausible scenario); 470 pCi/g for Scenario C (resident farmer using pond water as the only water source: a possible but unlikely scenario); and 120 pCi/g for Scenario D (resident farmer using well water as the only water source: a possible but unlikely scenario). The uranium guideline applies to the total activity concentration of uranium isotopes in their natural activity concentration ratio of 1:1: 0.046. These guidelines are calculated on the basis of a dose of 100 mrem/yr. In setting the actual uranium guideline for the Elza Gate Site, the DOE will apply the as low as reasonably achievable (ALARA) policy to the decision-making process, along with other factors, such as determining whether a particular scenario is reasonable and appropriate. 10 refs., 3 figs., 7 tabs

  10. Role of a cysteine residue in the active site of ERK and the MAPKK family

    International Nuclear Information System (INIS)

    Ohori, Makoto; Kinoshita, Takayoshi; Yoshimura, Seiji; Warizaya, Masaichi; Nakajima, Hidenori; Miyake, Hiroshi

    2007-01-01

    Kinases of mitogen-activated protein kinase (MAPK) cascades, including extracellular signal-regulated protein kinase (ERK), represent likely targets for pharmacological intervention in proliferative diseases. Here, we report that FR148083 inhibits ERK2 enzyme activity and TGFβ-induced AP-1-dependent luciferase expression with respective IC 50 values of 0.08 and 0.05 μM. FR265083 (1'-2' dihydro form) and FR263574 (1'-2' and 7'-8' tetrahydro form) exhibited 5.5-fold less and no activity, respectively, indicating that both the α,β-unsaturated ketone and the conformation of the lactone ring contribute to this inhibitory activity. The X-ray crystal structure of the ERK2/FR148083 complex revealed that the compound binds to the ATP binding site of ERK2, involving a covalent bond to Sγ of ERK2 Cys166, hydrogen bonds with the backbone NH of Met108, Nζ of Lys114, backbone C=O of Ser153, Nδ2 of Asn154, and hydrophobic interactions with the side chains of Ile31, Val39, Ala52, and Leu156. The covalent bond motif in the ERK2/FR148083 complex assures that the inhibitor has high activity for ERK2 and no activity for other MAPKs such as JNK1 and p38MAPKα/β/γ/δ which have leucine residues at the site corresponding to Cys166 in ERK2. On the other hand, MEK1 and MKK7, kinases of the MAPKK family which also can be inhibited by FR148083, contain a cysteine residue corresponding to Cys166 of ERK2. The covalent binding to the common cysteine residue in the ATP-binding site is therefore likely to play a crucial role in the inhibitory activity for these MAP kinases. These findings on the molecular recognition mechanisms of FR148083 for kinases with Cys166 should provide a novel strategy for the pharmacological intervention of MAPK cascades

  11. Biosynthesis of GDP-fucose and other sugar nucleotides in the blood stages of Plasmodium falciparum.

    Science.gov (United States)

    Sanz, Sílvia; Bandini, Giulia; Ospina, Diego; Bernabeu, Maria; Mariño, Karina; Fernández-Becerra, Carmen; Izquierdo, Luis

    2013-06-07

    Carbohydrate structures play important roles in many biological processes, including cell adhesion, cell-cell communication, and host-pathogen interactions. Sugar nucleotides are activated forms of sugars used by the cell as donors for most glycosylation reactions. Using a liquid chromatography-tandem mass spectrometry-based method, we identified and quantified the pools of UDP-glucose, UDP-galactose, UDP-N-acetylglucosamine, GDP-mannose, and GDP-fucose in Plasmodium falciparum intraerythrocytic life stages. We assembled these data with the in silico functional reconstruction of the parasite metabolic pathways obtained from the P. falciparum annotated genome, exposing new active biosynthetic routes crucial for further glycosylation reactions. Fucose is a sugar present in glycoconjugates often associated with recognition and adhesion events. Thus, the GDP-fucose precursor is essential in a wide variety of organisms. P. falciparum presents homologues of GDP-mannose 4,6-dehydratase and GDP-L-fucose synthase enzymes that are active in vitro, indicating that most GDP-fucose is formed by a de novo pathway that involves the bioconversion of GDP-mannose. Homologues for enzymes involved in a fucose salvage pathway are apparently absent in the P. falciparum genome. This is in agreement with in vivo metabolic labeling experiments showing that fucose is not significantly incorporated by the parasite. Fluorescence microscopy of epitope-tagged versions of P. falciparum GDP-mannose 4,6-dehydratase and GDP-L-fucose synthase expressed in transgenic 3D7 parasites shows that these enzymes localize in the cytoplasm of P. falciparum during the intraerythrocytic developmental cycle. Although the function of fucose in the parasite is not known, the presence of GDP-fucose suggests that the metabolite may be used for further fucosylation reactions.

  12. Biosynthesis of GDP-fucose and Other Sugar Nucleotides in the Blood Stages of Plasmodium falciparum*

    Science.gov (United States)

    Sanz, Sílvia; Bandini, Giulia; Ospina, Diego; Bernabeu, Maria; Mariño, Karina; Fernández-Becerra, Carmen; Izquierdo, Luis

    2013-01-01

    Carbohydrate structures play important roles in many biological processes, including cell adhesion, cell-cell communication, and host-pathogen interactions. Sugar nucleotides are activated forms of sugars used by the cell as donors for most glycosylation reactions. Using a liquid chromatography-tandem mass spectrometry-based method, we identified and quantified the pools of UDP-glucose, UDP-galactose, UDP-N-acetylglucosamine, GDP-mannose, and GDP-fucose in Plasmodium falciparum intraerythrocytic life stages. We assembled these data with the in silico functional reconstruction of the parasite metabolic pathways obtained from the P. falciparum annotated genome, exposing new active biosynthetic routes crucial for further glycosylation reactions. Fucose is a sugar present in glycoconjugates often associated with recognition and adhesion events. Thus, the GDP-fucose precursor is essential in a wide variety of organisms. P. falciparum presents homologues of GDP-mannose 4,6-dehydratase and GDP-l-fucose synthase enzymes that are active in vitro, indicating that most GDP-fucose is formed by a de novo pathway that involves the bioconversion of GDP-mannose. Homologues for enzymes involved in a fucose salvage pathway are apparently absent in the P. falciparum genome. This is in agreement with in vivo metabolic labeling experiments showing that fucose is not significantly incorporated by the parasite. Fluorescence microscopy of epitope-tagged versions of P. falciparum GDP-mannose 4,6-dehydratase and GDP-l-fucose synthase expressed in transgenic 3D7 parasites shows that these enzymes localize in the cytoplasm of P. falciparum during the intraerythrocytic developmental cycle. Although the function of fucose in the parasite is not known, the presence of GDP-fucose suggests that the metabolite may be used for further fucosylation reactions. PMID:23615908

  13. Rebaselining of the plutonium residue elimination project at Rocky Flats Environmental Technology Site

    International Nuclear Information System (INIS)

    Sailor, W.C.; Catlett, D.S.; Burns, T.P.

    1997-01-01

    Systems Engineering and Value Engineering principles were put into practice in rebaselining the Pu Residue Stabilization and Elimination Project at the Rocky Flats Environmental Technology Site. Tradeoff studies were conducted as to how to best rebaseline the system under the new Safeguards Termination Limits (STSs) issued by the Department of Energy. Through the use of a computerized database, the means by which Stakeholder values and other high-level requirements have been included in the tradeoff studies were documented. 13 refs., 2 figs., 1 tab

  14. Rebaselining of the plutonium residue elimination project at Rocky Flats Environmental Technology Site

    Energy Technology Data Exchange (ETDEWEB)

    Sailor, W.C.; Catlett, D.S.; Burns, T.P. [and others

    1997-03-01

    Systems Engineering and Value Engineering principles were put into practice in rebaselining the Pu Residue Stabilization and Elimination Project at the Rocky Flats Environmental Technology Site. Tradeoff studies were conducted as to how to best rebaseline the system under the new Safeguards Termination Limits (STSs) issued by the Department of Energy. Through the use of a computerized database, the means by which Stakeholder values and other high-level requirements have been included in the tradeoff studies were documented. 13 refs., 2 figs., 1 tab.

  15. A C. elegans-based foam for rapid on-site detection of residual live virus.

    Energy Technology Data Exchange (ETDEWEB)

    Negrete, Oscar A.; Branda, Catherine; Hardesty, Jasper O. E. (Sandia National Laboratories, Albuquerque, NM); Tucker, Mark David (Sandia National Laboratories, Albuquerque, NM); Kaiser, Julia N. (Global Product Management, Hilden, Germany); Kozina, Carol L.; Chirica, Gabriela S.

    2012-02-01

    In the response to and recovery from a critical homeland security event involving deliberate or accidental release of biological agents, initial decontamination efforts are necessarily followed by tests for the presence of residual live virus or bacteria. Such 'clearance sampling' should be rapid and accurate, to inform decision makers as they take appropriate action to ensure the safety of the public and of operational personnel. However, the current protocol for clearance sampling is extremely time-intensive and costly, and requires significant amounts of laboratory space and capacity. Detection of residual live virus is particularly problematic and time-consuming, as it requires evaluation of replication potential within a eukaryotic host such as chicken embryos. The intention of this project was to develop a new method for clearance sampling, by leveraging Sandia's expertise in the biological and material sciences in order to create a C. elegans-based foam that could be applied directly to the entire contaminated area for quick and accurate detection of any and all residual live virus by means of a fluorescent signal. Such a novel technology for rapid, on-site detection of live virus would greatly interest the DHS, DoD, and EPA, and hold broad commercial potential, especially with regard to the transportation industry.

  16. Cloning, Site-Directed Mutagenesis, and Functional Analysis of Active Residues in Lymantria dispar Chitinase.

    Science.gov (United States)

    Fan, Xiao-Jun; Yang, Chun; Zhang, Chang; Ren, Hui; Zhang, Jian-Dong

    2018-01-01

    Chitinases are glycosyl hydrolases that catalyze the hydrolysis of β-(1,4)-glycosidic bonds in chitin, the major structural polysaccharide presented in the cuticle and gut peritrophic matrix of insects. Two aspartate residues (D143, D145) and one tryptophan (W146) in the Lymantria dispar chitinase are highly conserved residues observed within the second conserved motif of the family 18 chitinase catalytic region. In this study, a chitinase cDNA, LdCht5, was cloned from L. dispar, and the roles of the three residues were investigated using site-directed mutagenesis and substituting them with three other amino acids. Seven mutant proteins, D143E, D145E, W146G, D143E/D145E, D143E/W146G, D145E/W146G, and D143E/D145E/W146G, as well as the wild-type enzyme, were produced using the baculovirus-insect cell line expression system. The enzymatic and kinetic properties of these mutant enzymes were measured using the oligosaccharide substrate MU-(GlcNAc) 3 . Among the seven mutants, the D145E, D143E/D145E, and D145E/W146G mutations kept some extant catalytic activity toward MU-(GlcNAc) 3 , while the D143E, W146G, D143E/W146G, and D143E/D145E/W146G mutant enzymes were inactivated. Compared with the mutant enzymes, the wild-type enzyme had higher values of k cat and k cat / K m . A study of the multiple point mutations in the second conserved catalytic region would help to elucidate the role of the critical residues and their relationships.

  17. Testing in support of on-site storage of residues in the Pipe Overpack Container

    International Nuclear Information System (INIS)

    Ammerman, D.J.; Bobbe, J.G.; Arviso, M.

    1997-02-01

    The disposition of the large back-log of plutonium residues at the Rocky Flats Environmental Technology Site (Rocky Flats) will require interim storage and subsequent shipment to a waste repository. Current plans call for disposal at the Waste Isolation Pilot Plant (WIPP) and the transportation to WIPP in the TRUPACT-II. The transportation phase will require the residues to be packaged in a container that is more robust than a standard 55-gallon waste drum. Rocky Flats has designed the Pipe Overpack Container to meet this need. It is desirable to use this same waste packaging for interim on-site storage in non-hardened buildings. To meet the safety concerns for this storage the Pipe Overpack Container has been subjected to a series of tests at Sandia National Laboratories in Albuquerque, New Mexico. In addition to the tests required to qualify the Pipe Overpack Container as a waste container for shipment in the TRUPACT-II several tests were performed solely for the purpose of qualifying the container for interim storage. This report will describe these tests and the packages response to the tests. 12 figs., 3 tabs

  18. A radiographic study on the prevalence of knife-edge residual alveolar ridge at proposed dental implant sites

    International Nuclear Information System (INIS)

    AlFaleh, Wafaa

    2009-01-01

    Dental implants are widely used in restoration of completely or partially edentulous dental arches. Before placement of endosseous implants in the jaws, both the quantity and quality of the residual ridge must be assessed radiographically. Remodeling activity after tooth extraction is localized primarily at the crestal area of the residual ridges, resulting in reduction of the height of bone and creation of various three-dimensional shapes of the residual ridges. When bone resorption at the lingual and buccal aspects is greater than that at the crestal area, a knife-edge type of residual ridge develops. The aim of this study was to evaluate the prevalence of the knife-edge morphology of the residual alveolar bone at proposed implant sites in partially or completely edentulous patients. Computed tomography (CT) cross-sectional images of the upper and lower jaws were assessed at the proposed sites before implant placement. Images of 258 proposed implant sites belonging to 30 patients were assessed radiographically. In 120 proposed implant sites out of 258 (46.5%), the residual alveolar ridge had a knife-edge configuration, the majority belonging to completely edentulous patients who lost their teeth more than ten years previously. High prevalence of knife-edge ridge was found, therefore, replacement of missing teeth by immediate implant is recommended to prevent atrophy or knife-edge morphology of the residual ridge. (author)

  19. Modulation of GDP-fucose level for generating proteins with reduced rate of fucosylation (WO2010141855).

    Science.gov (United States)

    Taupin, Philippe

    2011-09-01

    The application (WO2010141855) is in the field of glycobiology, and involves the control of the rate of fucosylation of proteins by exogenous factors. It aims at controlling the rate of protein fucosylation with inhibitors (drugs or nucleic acid antagonists) of enzymes involved in the synthesis of GDP-fucose. Mammalian cell lines were cultured in the presence of inhibitors, for example, siRNA. The rates of GDP-fucose in cells and during protein fucosylation were characterized. The level of protein fucosylation decreases rapidly in response to a decrease in GDP-fucose level. The relationship between the rate of fucosylation of proteins and the level of GDP-fucose in a cell is non-linear. Reduction in the rate of protein fucosylation can be achieved with a minimal reduction of the level of GDP-fucose in cells. The paradigm may be used to synthesize proteins and antibodies, with a reduced rate of fucosylation. The application claims that the use of drugs or nucleic acid antagonists that inhibit the enzymes involved in GDP-fucose biosynthesis optimizes the level of GDP-fucose present in cells, and reduces the rate of fucosylation of glycoproteins.

  20. Core-6 fucose and the oligomerization of the 1918 pandemic influenza viral neuraminidase

    International Nuclear Information System (INIS)

    Wu, Zhengliang L.; Zhou, Hui; Ethen, Cheryl M.; Reinhold, Vernon N.

    2016-01-01

    The 1918 H1N1 influenza virus was responsible for one of the most deadly pandemics in human history. Yet to date, the structure component responsible for its virulence is still a mystery. In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. In this report, the N-glycans from both forms were released and characterized by mass spectrometry. It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. Even more surprisingly, the stalk region of the active form was almost completely devoid of core-6-linked fucose. These findings were further supported by the results obtained from in vitro incorporation of azido fucose and "3H-labeled fucose using core-6 fucosyltransferase, FUT8. In addition, the incorporation of fucose did not change the enzymatic activity of the active form, implying that core-6 fucose is not directly involved in the enzymatic activity. It is postulated that core-6 fucose prohibits the oligomerization and subsequent activation of the enzyme. - Graphical abstract: Proposed mechanism for how core-fucose prohibits the tetramerization of the 1918 pandemic viral neuraminidase. Only the cross section of the stalk region with two N-linked glycans are depicted for clarity. (A) Carbohydrate–carbohydrate interaction on non-fucosylated monomer allows tetramerization. (B) Core-fucosylation disrupts the interaction and prevents the tetramerization. - Highlights: • Expressed 1918 pandemic influenza viral neuraminidase has inactive and active forms. • The inactive form contains high level of core-6 fucose, while the active form lacks such modification. • Core fucose could interfere the oligomerization of the neuraminidase and thus its activation. • This discovery may explain why

  1. Core-6 fucose and the oligomerization of the 1918 pandemic influenza viral neuraminidase

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Zhengliang L., E-mail: Leon.wu@bio-techne.com [Bio-Techne Inc., 614 McKinley Place NE, Minneapolis, MN 55413 (United States); Zhou, Hui [Gregg Hall, UNH Glycomics Center, University of New Hampshire (United States); Ethen, Cheryl M. [Bio-Techne Inc., 614 McKinley Place NE, Minneapolis, MN 55413 (United States); Reinhold, Vernon N., E-mail: Vernon.Reinhold@unh.edu [Gregg Hall, UNH Glycomics Center, University of New Hampshire (United States)

    2016-04-29

    The 1918 H1N1 influenza virus was responsible for one of the most deadly pandemics in human history. Yet to date, the structure component responsible for its virulence is still a mystery. In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. In this report, the N-glycans from both forms were released and characterized by mass spectrometry. It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. Even more surprisingly, the stalk region of the active form was almost completely devoid of core-6-linked fucose. These findings were further supported by the results obtained from in vitro incorporation of azido fucose and {sup 3}H-labeled fucose using core-6 fucosyltransferase, FUT8. In addition, the incorporation of fucose did not change the enzymatic activity of the active form, implying that core-6 fucose is not directly involved in the enzymatic activity. It is postulated that core-6 fucose prohibits the oligomerization and subsequent activation of the enzyme. - Graphical abstract: Proposed mechanism for how core-fucose prohibits the tetramerization of the 1918 pandemic viral neuraminidase. Only the cross section of the stalk region with two N-linked glycans are depicted for clarity. (A) Carbohydrate–carbohydrate interaction on non-fucosylated monomer allows tetramerization. (B) Core-fucosylation disrupts the interaction and prevents the tetramerization. - Highlights: • Expressed 1918 pandemic influenza viral neuraminidase has inactive and active forms. • The inactive form contains high level of core-6 fucose, while the active form lacks such modification. • Core fucose could interfere the oligomerization of the neuraminidase and thus its activation. • This discovery may explain

  2. Derivation of guidelines for uranium residual radioactive material in soil at the New Brunswick Site, Middlesex County, New Jersey

    International Nuclear Information System (INIS)

    Dunning, D.; Kamboj, S.; Nimmagadda, M.; Yu, C.

    1996-02-01

    Residual radioactive material guidelines for uranium in soil were derived for the New Brunswick Site, located in Middlesex County, New Jersey. This site has been designated for remedial action under the Formerly Utilized Sites Remedial Action Program of the US Department of Energy (DOE). Residual radioactive material guidelines for individual radionuclides of concern and total uranium were derived on the basis of the requirement that the 50-year committed effective dose equivalent to a hypothetical individual who lives or works in the immediate vicinity of the New Brunswick Site should not exceed a dose of 30 mrem/yr following remedial action for the current-use and likely future-use scenarios or a dose of 100 mrem/yr for less likely future-use scenarios. The DOE residual radioactive material guideline computer code, RESRAD, was used in this evaluation; RESRAD implements the methodology described in the DOE manual for establishing residual radioactive material guidelines. The guidelines derived in this report are intended to apply to the remediation of these remaining residual radioactive materials at the site. The primary radionuclides of concern in these remaining materials are expected to be radium-226 and, to a lesser extent, natural uranium and thorium. The DOE has established generic cleanup guidelines for radium and thorium in soil; however, cleanup guidelines for other radionuclides must be derived on a site-specific basis

  3. The behaviour of residual contaminants at a former station site, Antarctica

    International Nuclear Information System (INIS)

    Webster, Jenny; Webster, Kerry; Nelson, Peter; Waterhouse, Emma

    2003-01-01

    Minor contamination by metals, phosphorus, and fuel products were found at a former research station site in Antarctica. - In 1994, New Zealand's only mainland Antarctic base, Vanda Station, was removed from the shores of Lake Vanda, in the McMurdo Dry Valleys region of southern Victoria Land, Antarctica. Residual chemical contamination of the station site has been identified, in the form of discrete fuel spills, locally elevated Pb, Zn, Ag and Cd concentrations in soil and elevated Cu, Ni, Co and phosphate concentrations in suprapermafrost fluids in a gully formerly used for domestic washing water disposal. Pathways for contaminant transfer to Lake Vanda, potential environmental impacts and specific remediation/monitoring options are considered. While some contaminants (particularly Zn) could be selectively leached from flooded soil, during a period of rising lake level, the small area of contaminated soils exposed and low level of contamination suggests that this would not adversely affect either shallow lake water quality or the growth of cyanobacteria. Phosphate-enhanced growth of the latter may, however, be a visible consequence of the minor contamination occurring at this site

  4. Analysis and radiological assessment of residues containing NORM materials resulting from earlier activities including modelling of typical industrial residues. Pt. 1. Historical investigation of the radiological relevance of NORM residues and concepts for site identification

    International Nuclear Information System (INIS)

    Reichelt, Andreas; Niedermayer, Matthias; Sitte, Beate; Hamel, Peter Michael

    2007-01-01

    Natural radionuclides are part of the human environment and of the raw materials used. Technical processes may cause their accumulation in residues, and the result will be so-called NORM materials (Naturally occurring radioactive material). The amended Radiation Protection Ordinance (StrlSchV 2001) specifies how the public should be protected, but there are also residues dating back before the issuing of the StrlSchV 2001, the so-called NORM residues. The project intended to assess the risks resulting from these residues. It comprises four parts. Part 1 was for clarification of the radiological relevance of NORM residues and for the development of concepts to detect them. The criterion for their radiological relevance was their activity per mass unit and the material volume accumulated through the centuries. The former was calculated from a wide bibliographic search in the relevant literature on radiation protection, while the mass volume was obtained by a detailed historical search of the consumption of materials that may leave NORM residues. These are, in particular, residues from coal and ore mining and processing. To identify concrete sites, relevant data sources were identified, and a concept for identification of concrete NORM residues was developed on this basis. (orig.) [de

  5. Measurement of residual CO2 saturation at a geological storage site using hydraulic tests

    Science.gov (United States)

    Rötting, T. S.; Martinez-Landa, L.; Carrera, J.; Russian, A.; Dentz, M.; Cubillo, B.

    2012-12-01

    Estimating long term capillary trapping of CO2 in aquifers remains a key challenge for CO2 storage. Zhang et al. (2011) proposed a combination of thermal, tracer, and hydraulic experiments to estimate the amount of CO2 trapped in the formation after a CO2 push and pull test. Of these three types of experiments, hydraulic tests are the simplest to perform and possibly the most informative. However, their potential has not yet been fully exploited. Here, a methodology is presented to interpret these tests and analyze which parameters can be estimated. Numerical and analytical solutions are used to simulate a continuous injection in a porous medium where residual CO2 has caused a reduction in hydraulic conductivity and an increase in storativity over a finite thickness (a few meters) skin around the injection well. The model results are interpreted using conventional pressure build-up and diagnostic plots (a plot of the drawdown s and the logarithmic derivative d s / d ln t of the drawdown as a function of time). The methodology is applied using the hydraulic parameters estimated for the Hontomin site (Northern Spain) where a Technology Demonstration Plant (TDP) for geological CO2 storage is planned to be set up. The reduction of hydraulic conductivity causes an increase in observed drawdowns, the increased storativity in the CO2 zone causes a delay in the drawdown curve with respect to the reference curve measured before CO2 injection. The duration (characteristic time) of these effects can be used to estimate the radius of the CO2 zone. The effects of reduced permeability and increased storativity are well separated from wellbore storage and natural formation responses, even if the CO2-brine interface is inclined (i.e. the CO2 forms a cone around the well). We find that both skin hydraulic conductivity and storativity (and thus residual CO2 saturation) can be obtained from the water injection test provided that water flow rate is carefully controlled and head build

  6. Derivation of guidelines for uranium residual radioactive material in soil at the Colonie Site, Colonie, New York

    International Nuclear Information System (INIS)

    Dunning, D.

    1996-05-01

    Residual radioactive material guidelines for uranium in soil were derived for the Colonie site located in Colonie, New York. This site has been designated for remedial action under the Formerly Utilized Sites Remedial Action Program (FUSRAP) of the U.S. Department of Energy (DOE). The site became contaminated with radioactive material as a result of operations conducted by National Lead (NL) Industries from 1958 to 1984; these activities included brass foundry operations, electroplating of metal products, machining of various components using depleted uranium, and limited work with small amounts of enriched uranium and thorium. The Colonie site comprises the former NL Industries property, now designated the Colonie Interim Storage Site (CISS), and 56 vicinity properties contaminated by fallout from airborne emissions; 53 of the vicinity properties were previously remediated between 1984 and 1988. In 1984, DOE accepted ownership of the CISS property from NL Industries. Residual radioactive material guidelines for individual radionuclides and total uranium were derived on the basis of the requirement that the 50-year committed effective dose equivalent to a hypothetical individual who lives or works in the immediate vicinity of the site should not exceed a dose of 30 mrem/yr following remedial action for the current use and likely future use scenarios or a dose of 100 mrem/yr for less likely future use scenarios. The DOE residual radioactive material guideline computer code, RESRAD, was used in this evaluation; RESRAD implements the methodology described in the DOE manual for establishing residual radioactive material guidelines

  7. Structures of NodZ α1,6-fucosyltransferase in complex with GDP and GDP-fucose.

    Science.gov (United States)

    Brzezinski, Krzysztof; Dauter, Zbigniew; Jaskolski, Mariusz

    2012-02-01

    Rhizobial NodZ α1,6-fucosyltransferase (α1,6-FucT) catalyzes the transfer of the fucose (Fuc) moiety from guanosine 5'-diphosphate-β-L-fucose to the reducing end of the chitin oligosaccharide core during Nod-factor (NF) biosynthesis. NF is a key signalling molecule required for successful symbiosis with a legume host for atmospheric nitrogen fixation. To date, only two α1,6-FucT structures have been determined, both without any donor or acceptor molecule that could highlight the structural background of the catalytic mechanism. Here, the first crystal structures of α1,6-FucT in complex with its substrate GDP-Fuc and with GDP, which is a byproduct of the enzymatic reaction, are presented. The crystal of the complex with GDP-Fuc was obtained through soaking of native NodZ crystals with the ligand and its structure has been determined at 2.35 Å resolution. The fucose residue is exposed to solvent and is disordered. The enzyme-product complex crystal was obtained by cocrystallization with GDP and an acceptor molecule, penta-N-acetyl-L-glucosamine (penta-NAG). The structure has been determined at 1.98 Å resolution, showing that only the GDP molecule is present in the complex. In both structures the ligands are located in a cleft formed between the two domains of NodZ and extend towards the C-terminal domain, but their conformations differ significantly. The structures revealed that residues in three regions of the C-terminal domain, which are conserved among α1,2-, α1,6- and protein O-fucosyltransferases, are involved in interactions with the sugar-donor molecule. There is also an interaction with the side chain of Tyr45 in the N-terminal domain, which is very unusual for a GT-B-type glycosyltransferase. Only minor conformational changes of the protein backbone are observed upon ligand binding. The only exception is a movement of the loop located between strand βC2 and helix αC3. In addition, there is a shift of the αC3 helix itself upon GDP

  8. Networks of High Mutual Information Define the Structural Proximity of Catalytic Sites: Implications for Catalytic Residue Identification

    DEFF Research Database (Denmark)

    Buslje, Cristina Marino; Teppa, Elin; Di Doménico, Tomas

    2010-01-01

    other non-functional conserved residues. Using a data set of 434 Pfam families included in the catalytic site atlas (CSA) database, we tested this hypothesis and demonstrated that MI can complement amino acid conservation scores to detect CR. The Kullback-Leibler (KL) conservation measurement was shown.......90, the Cls method was found to have a sensitivity of 0.816. In summary, we demonstrate that networks of residues with high MI provide a distinct signature on CR and propose that such a signature should be present in other classes of functional residues where the requirement to maintain a particular function...

  9. Involvement of a bacterial microcompartment in the metabolism of fucose and rhamnose by Clostridium phytofermentans.

    Directory of Open Access Journals (Sweden)

    Elsa Petit

    Full Text Available Clostridium phytofermentans, an anaerobic soil bacterium, can directly convert plant biomass into biofuels. The genome of C. phytofermentans contains three loci with genes encoding shell proteins of bacterial microcompartments (BMC, organelles composed entirely of proteins.One of the BMC loci has homology to a BMC-encoding locus implicated in the conversion of fucose to propanol and propionate in a human gut commensal, Roseburia inulinivorans. We hypothesized that it had a similar role in C. phytofermentans. When C. phytofermentans was grown on fucose, the major products identified were ethanol, propanol and propionate. Transmission electron microscopy of fucose- and rhamnose-grown cultures revealed polyhedral structures, presumably BMCs. Microarray analysis indicated that during growth on fucose, operons coding for the BMC locus, fucose dissimilatory enzymes, and an ATP-binding cassette transporter became the dominant transcripts. These data are consistent with fucose fermentation producing a 1,2-propanediol intermediate that is further metabolized in the microcompartment encoded in the BMC locus. Growth on another deoxyhexose sugar, rhamnose, resulted in the expression of the same BMC locus and similar fermentation products. However, a different set of dissimilatory enzymes and transport system genes were induced. Quite surprisingly, growth on fucose or rhamnose also led to the expression of a diverse array of complex plant polysaccharide-degrading enzymes.Based on physiological, genomic, and microarray analyses, we propose a model for the fermentation of fucose and rhamnose in C. phytofermentans that includes enzymes encoded in the same BMC locus. Comparative genomic analysis suggests that this BMC may be present in other clostridial species.

  10. Site-directed Mutagenesis of Cysteine Residues in Phi-class Glutathione S-transferase F3 from Oryza sativa

    Energy Technology Data Exchange (ETDEWEB)

    Jo, Hyunjoo; Lee, Juwon; Noh, Jinseok; Kong, Kwanghoon [Chung-Ang Univ., Seoul (Korea, Republic of)

    2012-12-15

    To elucidate the roles of cysteine residues in rice Phi-class GST F3, in this study, all three cysteine residues were replaced with alanine by site-directed mutagenesis in order to obtain mutants C22A, C73A and C77A. Three mutant enzymes were expressed in Escherichia coli and purified to electrophoretic homogeneity by affinity chromatography on immobilized GSH. The substitutions of Cys73 and Cys77 residues in OsGSTF3 with alanine did not affect the glutathione conjugation activities, showing non-essentiality of these residues. On the other hand, the substitution of Cys22 residue with alanine resulted in approximately a 60% loss of specific activity toward ethacrynic acid. Moreover, the K{sub m}{sup CDNB} value of the mutant C22A was approximately 2.2 fold larger than that of the wild type. From these results, the evolutionally conserved cysteine 22 residue seems to participate rather in the structural stability of the active site in OsGSTF3 by stabilizing the electrophilic substrates-binding site's conformation than in the substrate binding directly.

  11. Coevolving residues of (beta/alpha)(8)-barrel proteins play roles in stabilizing active site architecture and coordinating protein dynamics.

    Science.gov (United States)

    Shen, Hongbo; Xu, Feng; Hu, Hairong; Wang, Feifei; Wu, Qi; Huang, Qiang; Wang, Honghai

    2008-12-01

    Indole-3-glycerol phosphate synthase (IGPS) is a representative of (beta/alpha)(8)-barrel proteins-the most common enzyme fold in nature. To better understand how the constituent amino-acids work together to define the structure and to facilitate the function, we investigated the evolutionary and dynamical coupling of IGPS residues by combining statistical coupling analysis (SCA) and molecular dynamics (MD) simulations. The coevolving residues identified by the SCA were found to form a network which encloses the active site completely. The MD simulations showed that these coevolving residues are involved in the correlated and anti-correlated motions. The correlated residues are within van der Waals contact and appear to maintain the active site architecture; the anti-correlated residues are mainly distributed on opposite sides of the catalytic cavity and coordinate the motions likely required for the substrate entry and product release. Our findings might have broad implications for proteins with the highly conserved (betaalpha)(8)-barrel in assessing the roles of amino-acids that are moderately conserved and not directly involved in the active site of the (beta/alpha)(8)-barrel. The results of this study could also provide useful information for further exploring the specific residue motions for the catalysis and protein design based on the (beta/alpha)(8)-barrel scaffold.

  12. Allowable Residual Contamination Levels in soil for decommissioning the Shippingport Atomic Power Station site

    International Nuclear Information System (INIS)

    Kennedy, W.E. Jr.; Napier, B.A.; Soldat, J.K.

    1983-09-01

    As part of decommissioning the Shippingport Atomic Power Station, a fundamental concern is the determination of Allowable Residual Contamination Levels (ARCL) for radionuclides in the soil at the site. The ARCL method described in this report is based on a scenario/exposure-pathway analysis and compliance with an annual dose limit for unrestricted use of the land after decommissioning. In addition to naturally occurring radionuclides and fallout from weapons testing, soil contamination could potentially come from five other sources. These include operation of the Shippingport Station as a pressurized water reactor, operations of the Shippingport Station as a light-water breeder, operation of the nearby Beaver Valley reactors, releases during decommissioning, and operation of other nearby industries, including the Bruce-Mansfield coal-fired power plants. ARCL values are presented for 29 individual radionculides and a worksheet is provided so that ARCL values can be determined for any mixture of the individual radionuclides for any annual dose limit selected. In addition, a worksheet is provided for calculating present time soil concentration value that will decay to the ARCL values after any selected period of time, such as would occur during a period of restricted access. The ARCL results are presented for both unconfined (surface) and confined (subsurface) soil contamination. The ARCL method and results described in this report provide a flexible means of determining unrestricted-use site release conditions after decommissioning the Shippingport Atomic Power Station

  13. Residual radioactivity in the vicinity of formerly utilized MED/AEC sites

    International Nuclear Information System (INIS)

    Haywood, F.F.; Goldsmith, W.A.

    1980-01-01

    As demand for uranium and thorium was accelerated during the 1940's, services of chemical and metallurgical firms and major research facilities were contracted as needed by the Manhattan Engineer District (MED). A lack of documentation of the radiological status at the time contracts were terminated at these facilities led the Department of Energy (DOE), and its predecessor the Energy Research and Development Administration (ERDA), to develop a major radiological resurvey program to fill this information void. A combination of aerial and ground-level radiological monitoring teams were utilized to identify and assess off-site radioactivity. Results from comprehensive aerial surveys provide the approximate areal extent of elevated radiation levels on the ground. These aerial survey results led to two types of ground-level surveys: (1) gamma-ray scanning on foot or from a motorized vehicle (mobile lab based system) to pinpoint the location of residual radioactivity; and (2) compehensive radiological surveys to determine the amount and type of materials present on specific parcels of private and public property identified during the scanning. This type of investigation was initiated in 1978 and has been successful in identifying and assessing the potential radiation hazard from property on which materials bearing natural radioactivity have been found. This paper contains a description of the techniques used to find and evaluate radioactive material displaced outside the boundaries of a formerly utilized site

  14. Functional expression of a human GDP-L-fucose transporter in Escherichia coli.

    Science.gov (United States)

    Förster-Fromme, Karin; Schneider, Sarah; Sprenger, Georg A; Albermann, Christoph

    2017-02-01

    To investigate the translocation of nucleotide-activated sugars from the cytosol across a membrane into the endoplasmatic reticulum or the Golgi apparatus which is an important step in the synthesis of glycoproteins and glycolipids in eukaryotes. The heterologous expression of the recombinant and codon-adapted human GDP-L-fucose antiporter gene SLC35C1 (encoding an N-terminal OmpA-signal sequence) led to a functional transporter protein located in the cytoplasmic membrane of Escherichia coli. The in vitro transport was investigated using inverted membrane vesicles. SLC35C1 is an antiporter specific for GDP-L-fucose and depending on the concomitant reverse transport of GMP. The recombinant transporter FucT1 exhibited an activity for the transport of 3 H-GDP-L-fucose with a V max of 8 pmol/min mg with a K m of 4 µM. The functional expression of SLC35C1 in GDP-L-fucose overproducing E. coli led to the export of GDP-L-fucose to the culture supernatant. The export of GDP-L-fucose by E. coli provides the opportunity for the engineering of a periplasmatic fucosylation reaction in recombinant bacterial cells.

  15. L-fucose utilization provides Campylobacter jejuni with a competitive advantage.

    Science.gov (United States)

    Stahl, Martin; Friis, Lorna M; Nothaft, Harald; Liu, Xin; Li, Jianjun; Szymanski, Christine M; Stintzi, Alain

    2011-04-26

    Campylobacter jejuni is a prevalent gastrointestinal pathogen in humans and a common commensal of poultry. When colonizing its hosts, C. jejuni comes into contact with intestinal carbohydrates, including L-fucose, released from mucin glycoproteins. Several strains of C. jejuni possess a genomic island (cj0480c-cj0490) that is up-regulated in the presence of both L-fucose and mucin and allows for the utilization of L-fucose as a substrate for growth. Strains possessing this genomic island show increased growth in the presence of L-fucose and mutation of cj0481, cj0486, and cj0487 results in the loss of the ability to grow on this substrate. Furthermore, mutants in the putative fucose permease (cj0486) are deficient in fucose uptake and demonstrate a competitive disadvantage when colonizing the piglet model of human disease, which is not paralleled in the colonization of poultry. This identifies a previously unrecorded metabolic pathway in select strains of C. jejuni associated with a virulent lifestyle.

  16. Derivation of residual radioactive material guidelines for uranium in soil at the Middlesex Sampling Plant Site, Middlesex, New Jersey

    International Nuclear Information System (INIS)

    Dunning, D.E.

    1995-02-01

    Residual radioactive material guidelines for uranium in soil were derived for the Middlesex Sampling Plant (MSP) site in Middlesex, New Jersey. This site has been designated for remedial action under the Formerly Utilized Sites Remedial Action Program (FUSRAP) of the US Department of Energy. The site became contaminated from operations conducted in support of the Manhattan Engineer District (MED) and the Atomic Energy Commission (AEC) between 1943 and 1967. Activities conducted at the site included sampling, storage, and shipment of uranium, thorium, and beryllium ores and residues. Uranium guidelines for single radioisotopes and total uranium were derived on the basis of the requirement that the 50-year committed effective dose equivalent to a hypothetical individual living or working in the immediate vicinity of the MSP site should not exceed a dose of 30 mrem/yr following remedial action for the current-use and likely future-use scenarios or a dose of 100 mrem/yr for less likely future-use scenarios. The RESRAD computer code, which implements the methodology described in the DOE manual for establishing residual radioactive material guidelines, was used in this evaluation. Four scenarios were considered for the site. These scenarios vary regarding future land use at the site, sources of water used, and sources of food consumed

  17. Identification of critical residues in loop E in the 5-HT3ASR binding site

    Directory of Open Access Journals (Sweden)

    Muthalagi Mani

    2002-06-01

    Full Text Available Abstract Background The serotonin type 3 receptor (5-HT3R is a member of a superfamily of ligand gated ion channels. All members of this family share a large degree of sequence homology and presumably significant structural similarity. A large number of studies have explored the structure-function relationships of members of this family, particularly the nicotinic and GABA receptors. This information can be utilized to gain additional insights into specific structural and functional features of other receptors in this family. Results Thirteen amino acids in the mouse 5-HT3ASR that correspond to the putative E binding loop of the nicotinic α7 receptor were chosen for mutagenesis. Due to the presence of a highly conserved glycine in this region, it has been suggested that this binding loop is comprised of a hairpin turn and may form a portion of the ligand-binding site in this ion channel family. Mutation of the conserved glycine (G147 to alanine eliminated binding of the 5-HT3R antagonist [3H]granisetron. Three tyrosine residues (Y140, Y142 and Y152 also significantly altered the binding of 5-HT3R ligands. Mutations in neighboring residues had little or no effect on binding of these ligands to the 5-HT3ASR. Conclusion Our data supports a role for the putative E-loop region of the 5-HT3R in the binding of 5-HT, mCPBG, d-tc and lerisetron. 5-HT and mCPBG interact with Y142, d-tc with Y140 and lerisetron with both Y142 and Y152. Our data also provides support for the hypothesis that this region of the receptor is present in a loop structure.

  18. Development of an on-site measurement method for residual stress in primary system piping of nuclear power plants

    International Nuclear Information System (INIS)

    Maekawa, Akira; Takahashi, Shigeru; Fujiwara, Masaharu

    2014-01-01

    In residual stress measurement for large-scale pipes and vessels in high radiation areas and highly contaminated areas of nuclear plants, it is difficult to bring the radioactivated pipes and vessels out of the areas as they are. If they can brought out, it is very burdensome to handle them for the measurement. Development of an on-site measurement method of residual stress which can be quickly applied and has sufficient measurement accuracy is desirable. In this study, a new method combining an electric discharge skim-cut method with a microscopic strain measurement method using markers was proposed to realize the on-site residual stress measurement on pipes in high radiation areas and highly contaminated areas. In the electric discharge skim-cut method, a boat-type sample is skimmed out of a pipe outer/inner surface using electric discharge machining and released residual stress is measured. The on-site measurement of residual stress by the method can be done using a small, portable electric discharge machine. In the microscopic strain measurement method using markers, the residual stress is estimated by microscopic measurement of the distance between markers after the stress release. The combination of both methods can evaluate the residual stress with the same accuracy as conventional methods offer and it can achieve reduction of radiation exposure in the measurement because the work is done simply and rapidly. In this study, the applicability of the electric discharge skim-cut method was investigated because the applicability of the microscopic strain measurement method using markers was confirmed previously. The experimental examination clarified the applicable conditions for the residual stress measurement with the same accuracy as the conventional methods. Furthermore, the electric discharge machining conditions using pure water as the machining liquid was found to eliminate the amount of liquid radioactive waste completely. (author)

  19. On using residual risk to assess the cost effectiveness and health protectiveness of remedy selection at superfund sites

    International Nuclear Information System (INIS)

    Katsumata, Peter T.; Kastenberg, William E.

    1998-01-01

    This article examines the importance of determining residual risk and its impact on remedy selection at Superfund Sites. Within this examination, risks are assessed using probabilistic models that incorporate the uncertainty and variability of the input parameters, and utilize parameter distributions based on current and applicable site-specific data. Monte Carlo methods are used to propagate these uncertainties and variabilities through the risk calculations resulting in a distribution for the estimate of both risk and residual risk. Such an approach permits an informed decision based on a broad information base which involves considering the entire uncertainty distribution of risk rather than a point estimate for each exposure scenario. Using the probabilistic risk estimates, with current and applicable site-specific data, alternative decisions regarding cleanup are obtained for two Superfund Sites

  20. Molecular Basis for Enzymatic Sulfite Oxidation -- HOW THREE CONSERVED ACTIVE SITE RESIDUES SHAPE ENZYME ACTIVITY

    Energy Technology Data Exchange (ETDEWEB)

    Bailey, Susan; Rapson, Trevor; Johnson-Winters, Kayunta; Astashkin, Andrei; Enemark, John; Kappler, Ulrike

    2008-11-10

    Sulfite dehydrogenases (SDHs) catalyze the oxidation and detoxification of sulfite to sulfate, a reaction critical to all forms of life. Sulfite-oxidizing enzymes contain three conserved active site amino acids (Arg-55, His-57, and Tyr-236) that are crucial for catalytic competency. Here we have studied the kinetic and structural effects of two novel and one previously reported substitution (R55M, H57A, Y236F) in these residues on SDH catalysis. Both Arg-55 and His-57 were found to have key roles in substrate binding. An R55M substitution increased Km(sulfite)(app) by 2-3 orders of magnitude, whereas His-57 was required for maintaining a high substrate affinity at low pH when the imidazole ring is fully protonated. This effect may be mediated by interactions of His-57 with Arg-55 that stabilize the position of the Arg-55 side chain or, alternatively, may reflect changes in the protonation state of sulfite. Unlike what is seen for SDHWT and SDHY236F, the catalytic turnover rates of SDHR55M and SDHH57A are relatively insensitive to pH (~;;60 and 200 s-1, respectively). On the structural level, striking kinetic effects appeared to correlate with disorder (in SDHH57A and SDHY236F) or absence of Arg-55 (SDHR55M), suggesting that Arg-55 and the hydrogen bonding interactions it engages in are crucial for substrate binding and catalysis. The structure of SDHR55M has sulfate bound at the active site, a fact that coincides with a significant increase in the inhibitory effect of sulfate in SDHR55M. Thus, Arg-55 also appears to be involved in enabling discrimination between the substrate and product in SDH.

  1. The disruption of GDP-fucose de novo biosynthesis suggests the presence of a novel fucose-containing glycoconjugate in Plasmodium asexual blood stages.

    Science.gov (United States)

    Sanz, Sílvia; López-Gutiérrez, Borja; Bandini, Giulia; Damerow, Sebastian; Absalon, Sabrina; Dinglasan, Rhoel R; Samuelson, John; Izquierdo, Luis

    2016-11-16

    Glycosylation is an important posttranslational protein modification in all eukaryotes. Besides glycosylphosphatidylinositol (GPI) anchors and N-glycosylation, O-fucosylation has been recently reported in key sporozoite proteins of the malaria parasite. Previous analyses showed the presence of GDP-fucose (GDP-Fuc), the precursor for all fucosylation reactions, in the blood stages of Plasmodium falciparum. The GDP-Fuc de novo pathway, which requires the action of GDP-mannose 4,6-dehydratase (GMD) and GDP-L-fucose synthase (FS), is conserved in the parasite genome, but the importance of fucose metabolism for the parasite is unknown. To functionally characterize the pathway we generated a PfGMD mutant and analyzed its phenotype. Although the labelling by the fucose-binding Ulex europaeus agglutinin I (UEA-I) was completely abrogated, GDP-Fuc was still detected in the mutant. This unexpected result suggests the presence of an alternative mechanism for maintaining GDP-Fuc in the parasite. Furthermore, PfGMD null mutant exhibited normal growth and invasion rates, revealing that the GDP-Fuc de novo metabolic pathway is not essential for the development in culture of the malaria parasite during the asexual blood stages. Nonetheless, the function of this metabolic route and the GDP-Fuc pool that is generated during this stage may be important for gametocytogenesis and sporogonic development in the mosquito.

  2. The disruption of GDP-fucose de novo biosynthesis suggests the presence of a novel fucose-containing glycoconjugate in Plasmodium asexual blood stages

    Science.gov (United States)

    Sanz, Sílvia; López-Gutiérrez, Borja; Bandini, Giulia; Damerow, Sebastian; Absalon, Sabrina; Dinglasan, Rhoel R.; Samuelson, John; Izquierdo, Luis

    2016-01-01

    Glycosylation is an important posttranslational protein modification in all eukaryotes. Besides glycosylphosphatidylinositol (GPI) anchors and N-glycosylation, O-fucosylation has been recently reported in key sporozoite proteins of the malaria parasite. Previous analyses showed the presence of GDP-fucose (GDP-Fuc), the precursor for all fucosylation reactions, in the blood stages of Plasmodium falciparum. The GDP-Fuc de novo pathway, which requires the action of GDP-mannose 4,6-dehydratase (GMD) and GDP-L-fucose synthase (FS), is conserved in the parasite genome, but the importance of fucose metabolism for the parasite is unknown. To functionally characterize the pathway we generated a PfGMD mutant and analyzed its phenotype. Although the labelling by the fucose-binding Ulex europaeus agglutinin I (UEA-I) was completely abrogated, GDP-Fuc was still detected in the mutant. This unexpected result suggests the presence of an alternative mechanism for maintaining GDP-Fuc in the parasite. Furthermore, PfGMD null mutant exhibited normal growth and invasion rates, revealing that the GDP-Fuc de novo metabolic pathway is not essential for the development in culture of the malaria parasite during the asexual blood stages. Nonetheless, the function of this metabolic route and the GDP-Fuc pool that is generated during this stage may be important for gametocytogenesis and sporogonic development in the mosquito. PMID:27849032

  3. Regulatory Closure Options for the Residue in the Hanford Site Single-Shell Tanks

    International Nuclear Information System (INIS)

    Cochran, J.R.; Shyr, L.J.

    1998-01-01

    Liquid, mixed, high-level radioactive waste (HLW) has been stored in 149 single-shell tanks (SSTS) located in tank farms on the U.S. Department of Energy's (DOE's) Hanford Site. The DOE is developing technologies to retrieve as much remaining HLW as technically possible prior to physically closing the tank farms. In support of the Hanford Tanks Initiative, Sandia National Laboratories has addressed the requirements for the regulatory closure of the radioactive component of any SST residue that may remain after physical closure. There is significant uncertainty about the end state of each of the 149 SSTS; that is, the nature and amount of wastes remaining in the SSTS after retrieval is uncertain. As a means of proceeding in the face of these uncertainties, this report links possible end-states with associated closure options. Requirements for disposal of HLW and low-level radioactive waste (LLW) are reviewed in detail. Incidental waste, which is radioactive waste produced incidental to the further processing of HLW, is then discussed. If the low activity waste (LAW) fraction from the further processing of HLW is determined to be incidental waste, then DOE can dispose of that incidental waste onsite without a license from the U.S. Nuclear Regulatory Commissions (NRC). The NRC has proposed three Incidental Waste Criteria for determining if a LAW fraction is incidental waste. One of the three Criteria is that the LAW fraction should not exceed the NRC's Class C limits

  4. Regulatory Closure Options for the Residue in the Hanford Site Single-Shell Tanks

    Energy Technology Data Exchange (ETDEWEB)

    Cochran, J.R. Shyr, L.J.

    1998-10-05

    Liquid, mixed, high-level radioactive waste (HLW) has been stored in 149 single-shell tanks (SSTS) located in tank farms on the U.S. Department of Energy's (DOE's) Hanford Site. The DOE is developing technologies to retrieve as much remaining HLW as technically possible prior to physically closing the tank farms. In support of the Hanford Tanks Initiative, Sandia National Laboratories has addressed the requirements for the regulatory closure of the radioactive component of any SST residue that may remain after physical closure. There is significant uncertainty about the end state of each of the 149 SSTS; that is, the nature and amount of wastes remaining in the SSTS after retrieval is uncertain. As a means of proceeding in the face of these uncertainties, this report links possible end-states with associated closure options. Requirements for disposal of HLW and low-level radioactive waste (LLW) are reviewed in detail. Incidental waste, which is radioactive waste produced incidental to the further processing of HLW, is then discussed. If the low activity waste (LAW) fraction from the further processing of HLW is determined to be incidental waste, then DOE can dispose of that incidental waste onsite without a license from the U.S. Nuclear Regulatory Commissions (NRC). The NRC has proposed three Incidental Waste Criteria for determining if a LAW fraction is incidental waste. One of the three Criteria is that the LAW fraction should not exceed the NRC's Class C limits.

  5. Networks of high mutual information define the structural proximity of catalytic sites: implications for catalytic residue identification.

    Directory of Open Access Journals (Sweden)

    Cristina Marino Buslje

    Full Text Available Identification of catalytic residues (CR is essential for the characterization of enzyme function. CR are, in general, conserved and located in the functional site of a protein in order to attain their function. However, many non-catalytic residues are highly conserved and not all CR are conserved throughout a given protein family making identification of CR a challenging task. Here, we put forward the hypothesis that CR carry a particular signature defined by networks of close proximity residues with high mutual information (MI, and that this signature can be applied to distinguish functional from other non-functional conserved residues. Using a data set of 434 Pfam families included in the catalytic site atlas (CSA database, we tested this hypothesis and demonstrated that MI can complement amino acid conservation scores to detect CR. The Kullback-Leibler (KL conservation measurement was shown to significantly outperform both the Shannon entropy and maximal frequency measurements. Residues in the proximity of catalytic sites were shown to be rich in shared MI. A structural proximity MI average score (termed pMI was demonstrated to be a strong predictor for CR, thus confirming the proposed hypothesis. A structural proximity conservation average score (termed pC was also calculated and demonstrated to carry distinct information from pMI. A catalytic likeliness score (Cls, combining the KL, pC and pMI measures, was shown to lead to significantly improved prediction accuracy. At a specificity of 0.90, the Cls method was found to have a sensitivity of 0.816. In summary, we demonstrate that networks of residues with high MI provide a distinct signature on CR and propose that such a signature should be present in other classes of functional residues where the requirement to maintain a particular function places limitations on the diversification of the structural environment along the course of evolution.

  6. Networks of high mutual information define the structural proximity of catalytic sites: implications for catalytic residue identification.

    Science.gov (United States)

    Marino Buslje, Cristina; Teppa, Elin; Di Doménico, Tomas; Delfino, José María; Nielsen, Morten

    2010-11-04

    Identification of catalytic residues (CR) is essential for the characterization of enzyme function. CR are, in general, conserved and located in the functional site of a protein in order to attain their function. However, many non-catalytic residues are highly conserved and not all CR are conserved throughout a given protein family making identification of CR a challenging task. Here, we put forward the hypothesis that CR carry a particular signature defined by networks of close proximity residues with high mutual information (MI), and that this signature can be applied to distinguish functional from other non-functional conserved residues. Using a data set of 434 Pfam families included in the catalytic site atlas (CSA) database, we tested this hypothesis and demonstrated that MI can complement amino acid conservation scores to detect CR. The Kullback-Leibler (KL) conservation measurement was shown to significantly outperform both the Shannon entropy and maximal frequency measurements. Residues in the proximity of catalytic sites were shown to be rich in shared MI. A structural proximity MI average score (termed pMI) was demonstrated to be a strong predictor for CR, thus confirming the proposed hypothesis. A structural proximity conservation average score (termed pC) was also calculated and demonstrated to carry distinct information from pMI. A catalytic likeliness score (Cls), combining the KL, pC and pMI measures, was shown to lead to significantly improved prediction accuracy. At a specificity of 0.90, the Cls method was found to have a sensitivity of 0.816. In summary, we demonstrate that networks of residues with high MI provide a distinct signature on CR and propose that such a signature should be present in other classes of functional residues where the requirement to maintain a particular function places limitations on the diversification of the structural environment along the course of evolution.

  7. Enzyme active site mimics based on TriAzaCyclophane (TAC)-scaffolded peptides and amino acid residues

    NARCIS (Netherlands)

    Albada, H.B.

    2009-01-01

    This thesis describes the scope and limitations of the application of TriAzaCyclophane (TAC)-scaffolded peptides or amino acid residues as enzyme active site mimics, as ligands in asymmetric catalysis and as hydrolysis catalysts attached to vancomycin. For the mimicry of functional group enzymes, of

  8. A Simple Surface-Enhanced Raman Spectroscopic Method for on-Site Screening of Tetracycline Residue in Whole Milk

    Directory of Open Access Journals (Sweden)

    Sagar Dhakal

    2018-02-01

    Full Text Available Therapeutic and subtherapeutic use of veterinary drugs has increased the risk of residue contamination in animal food products. Antibiotics such as tetracycline are used for mastitis treatment of lactating cows. Milk expressed from treated cows before the withdrawal period has elapsed may contain tetracycline residue. This study developed a simple surface-enhanced Raman spectroscopic (SERS method for on-site screening of tetracycline residue in milk and water. Six batches of silver colloid nanoparticles were prepared for surface enhancement measurement. Milk-tetracycline and water-tetracycline solutions were prepared at seven concentration levels (1000, 500, 100, 10, 1, 0.1, and 0.01 ppm and spiked with silver colloid nanoparticles. A 785 nm Raman spectroscopic system was used for spectral measurement. Tetracycline vibrational modes were observed at 1285, 1317 and 1632 cm−1 in water-tetracycline solutions and 1322 and 1621 cm−1 (shifted from 1317 and 1632 cm−1, respectively in milk-tetracycline solutions. Tetracycline residue concentration as low as 0.01 ppm was detected in both the solutions. The peak intensities at 1285 and 1322 cm−1 were used to estimate the tetracycline concentrations in water and milk with correlation coefficients of 0.92 for water and 0.88 for milk. Results indicate that this SERS method is a potential tool that can be used on-site at field production for qualitative and quantitative detection of tetracycline residues.

  9. A Simple Surface-Enhanced Raman Spectroscopic Method for on-Site Screening of Tetracycline Residue in Whole Milk.

    Science.gov (United States)

    Dhakal, Sagar; Chao, Kuanglin; Huang, Qing; Kim, Moon; Schmidt, Walter; Qin, Jianwei; Broadhurst, C Leigh

    2018-02-01

    Therapeutic and subtherapeutic use of veterinary drugs has increased the risk of residue contamination in animal food products. Antibiotics such as tetracycline are used for mastitis treatment of lactating cows. Milk expressed from treated cows before the withdrawal period has elapsed may contain tetracycline residue. This study developed a simple surface-enhanced Raman spectroscopic (SERS) method for on-site screening of tetracycline residue in milk and water. Six batches of silver colloid nanoparticles were prepared for surface enhancement measurement. Milk-tetracycline and water-tetracycline solutions were prepared at seven concentration levels (1000, 500, 100, 10, 1, 0.1, and 0.01 ppm) and spiked with silver colloid nanoparticles. A 785 nm Raman spectroscopic system was used for spectral measurement. Tetracycline vibrational modes were observed at 1285, 1317 and 1632 cm -1 in water-tetracycline solutions and 1322 and 1621 cm -1 (shifted from 1317 and 1632 cm -1 , respectively) in milk-tetracycline solutions. Tetracycline residue concentration as low as 0.01 ppm was detected in both the solutions. The peak intensities at 1285 and 1322 cm -1 were used to estimate the tetracycline concentrations in water and milk with correlation coefficients of 0.92 for water and 0.88 for milk. Results indicate that this SERS method is a potential tool that can be used on-site at field production for qualitative and quantitative detection of tetracycline residues.

  10. SigniSite: Identification of residue-level genotype-phenotype correlations in protein multiple sequence alignments

    DEFF Research Database (Denmark)

    Jessen, Leon Ivar; Hoof, Ilka; Lund, Ole

    2013-01-01

    Site does not require any pre-definition of subgroups or binary classification. Input is a set of protein sequences where each sequence has an associated real number, quantifying a given phenotype. SigniSite will then identify which amino acid residues are significantly associated with the data set......) using a set of human immunodeficiency virus protease-inhibitor genotype–phenotype data and corresponding resistance mutation scores from the Stanford University HIV Drug Resistance Database, and a data set of protein families with experimentally annotated SDPs. For both data sets, SigniSite was found...

  11. Mutational and structural analyses of Caldanaerobius polysaccharolyticus Man5B reveal novel active site residues for family 5 glycoside hydrolases.

    Science.gov (United States)

    Oyama, Takuji; Schmitz, George E; Dodd, Dylan; Han, Yejun; Burnett, Alanna; Nagasawa, Naoko; Mackie, Roderick I; Nakamura, Haruki; Morikawa, Kosuke; Cann, Isaac

    2013-01-01

    CpMan5B is a glycoside hydrolase (GH) family 5 enzyme exhibiting both β-1,4-mannosidic and β-1,4-glucosidic cleavage activities. To provide insight into the amino acid residues that contribute to catalysis and substrate specificity, we solved the structure of CpMan5B at 1.6 Å resolution. The structure revealed several active site residues (Y12, N92 and R196) in CpMan5B that are not present in the active sites of other structurally resolved GH5 enzymes. Residue R196 in GH5 enzymes is thought to be strictly conserved as a histidine that participates in an electron relay network with the catalytic glutamates, but we show that an arginine fulfills a functionally equivalent role and is found at this position in every enzyme in subfamily GH5_36, which includes CpMan5B. Residue N92 is required for full enzymatic activity and forms a novel bridge over the active site that is absent in other family 5 structures. Our data also reveal a role of Y12 in establishing the substrate preference for CpMan5B. Using these molecular determinants as a probe allowed us to identify Man5D from Caldicellulosiruptor bescii as a mannanase with minor endo-glucanase activity.

  12. Mutational and structural analyses of Caldanaerobius polysaccharolyticus Man5B reveal novel active site residues for family 5 glycoside hydrolases.

    Directory of Open Access Journals (Sweden)

    Takuji Oyama

    Full Text Available CpMan5B is a glycoside hydrolase (GH family 5 enzyme exhibiting both β-1,4-mannosidic and β-1,4-glucosidic cleavage activities. To provide insight into the amino acid residues that contribute to catalysis and substrate specificity, we solved the structure of CpMan5B at 1.6 Å resolution. The structure revealed several active site residues (Y12, N92 and R196 in CpMan5B that are not present in the active sites of other structurally resolved GH5 enzymes. Residue R196 in GH5 enzymes is thought to be strictly conserved as a histidine that participates in an electron relay network with the catalytic glutamates, but we show that an arginine fulfills a functionally equivalent role and is found at this position in every enzyme in subfamily GH5_36, which includes CpMan5B. Residue N92 is required for full enzymatic activity and forms a novel bridge over the active site that is absent in other family 5 structures. Our data also reveal a role of Y12 in establishing the substrate preference for CpMan5B. Using these molecular determinants as a probe allowed us to identify Man5D from Caldicellulosiruptor bescii as a mannanase with minor endo-glucanase activity.

  13. Allowable residual-contamination levels for decommissioning facilities in the 100 areas of the Hanford Site

    International Nuclear Information System (INIS)

    Kennedy, W.E. Jr.; Napier, B.A.

    1983-07-01

    This report contains the results of a study sponsored by UNC Nuclear Industries to determine Allowable Residual Contamination Levels (ARCL) for five generic categories of facilities in the 100 Areas of the Hanford Site. The purpose of this study is to provide ARCL data useful to UNC engineers in conducting safety and cost comparisons for decommissioning alternatives. The ARCL results are based on a scenario/exposure-pathway analysis and compliance with an annual dose limit for three specific modes of future use of the land and facilities. These modes of use are restricted, controlled, and unrestricted. The information on ARCL values for restricted and controlled use provided by this report is intended to permit a full consideration of decommissioning alternatives. ARCL results are presented both for surface contamination remaining in facilities (in dpm/100 cm 2 ), and for unconfined surface and confined subsurface soil conditions (in pCi/g). Two confined soil conditions are considered: contamination at depths between 1 and 4 m, and contamination at depths greater than or equal to 5 m. A set of worksheets are presented in an appendix for modifying the ARCL values to accommodate changes in the radionuclide mixture or concentrations, to consider the impacts of radioactive decay, and to predict instrument responses. Finally, a comparison is made between the unrestricted release ARCL values for the 100 Area facilities and existing decommissioning and land disposal regulations. For surface contamination, the comparison shows good agreement. For soil contamination, the comparison shows good agreement if reasonable modification factors are applied to account for the differences in modeling soil contamination and licensed low-level waste

  14. Pneumatic transport system development: residuals and releases program at Westinghouse Cheswick site

    International Nuclear Information System (INIS)

    Larouere, P.J.; Shoulders, J.L.

    1979-01-01

    Plutonium oxide and uranium oxide powders are processed within glove boxes or within confinement systems during the fabrication of mixed oxide (MOX) pellets for recycle fuel. The release of these powders to the glove box or to the confinement results in some airborne material that is deposited in the enclosure or is carried in the air streams to the effluent air filtration system. Release tests on simulated leaks in pneumatic transport equipment and release tests on simulated failures with powder blending equipment were conducted. A task to develop pneumatic transport for the movement of powders within an MOX fabrication plant has been underway at the Westinghouse Research Laboratories. While testing and evaluating selected pneumatic transport components on a full scale were in progress, it was deemed necessary that final verification of the technology would have to be performed with plutonium-bearing powders because of the marked differences in certain properties of plutonium from those of uranium oxides. A smaller was designed and constructed for the planned installation in glove boxes at the Westinghouse Plutonium Fuel Development Laboratory. However, prior to use with plutonium it was agreed that this system be set up and tested with uranium oxide powder. The test program conducted at the Westinghouse Cheswick site was divided into two major parts. The first of these examined the residuals left as a result of the pneumatic transport of nuclear fuel powders and verified the operability of this one-third scale system. The second part of the program studied the amount of powder released to the air when off-standard process procedures or maintenance operations were conducted on the pneumatic transport system. Air samplers located within the walk-in box housing the transport loop were used to measure the solids concentration in the air. From this information, the total amount of airborne powder was determined

  15. Computation Of The Residual Radionuclide Activity Within Three Natural Waterways At The Savannah River Site

    Energy Technology Data Exchange (ETDEWEB)

    Hiergesell, R. A.; Phifer, M. A.

    2014-01-07

    In 2010 a Composite Analysis (CA) of the U.S. Department of Energy’s (DOE’s) Savannah River Site (SRS) was completed. This investigation evaluated the dose impact of the anticipated SRS End State residual sources of radionuclides to offsite members of the public. Doses were assessed at the locations where SRS site streams discharge into the Savannah River at the perimeter of the SRS. Although the model developed to perform this computation indicated that the dose constraint of 0.3 mSv/yr (30 mrem/yr), associated with CA, was not approached at the Points of Assessment (POAs), a significant contribution to the total computed dose was derived from the radionuclides (primarily Cs-137) bound-up in the soil and sediment of the drainage corridors of several SRS streams. DOE’s Low Level Waste Federal Review Group (LFRG) reviewed the 2010 CA and identified several items to be addressed in the SRS Maintenance Program. One of the items recognized Cs-137 in the Lower Three Runs (LTR) Integrator Operable Unit (IOU), as a significant CA dose driver. The item made the recommendation that SRS update the estimated radionuclide inventory, including Cs-137, in the LTR IOU. That initial work has been completed and its radionuclide inventory refined. There are five additional streams at SRS and the next phase of the response to the LFRG concern was to obtain a more accurate inventory and distribution of radionuclides in three of those streams, Fourmile Branch (FMB), Pen Branch (PB) and Steel Creek (SC). Each of these streams is designated as an IOU, which are defined for the purpose of this investigation as the surface water bodies and associated wetlands, including the channel sediment, floodplain sed/soil, and related biota. If present, radionuclides associated with IOUs are adsorbed to the streambed sediment and soils of the shallow floodplains that lie immediately adjacent to stream channels. The scope of this effort included the evaluation of any previous sampling and

  16. An L-Fucose Operon in the Probiotic Lactobacillus rhamnosus GG Is Involved in Adaptation to Gastrointestinal Conditions.

    Science.gov (United States)

    Becerra, Jimmy E; Yebra, María J; Monedero, Vicente

    2015-06-01

    L-Fucose is a sugar present in human secretions as part of human milk oligosaccharides, mucins, and other glycoconjugates in the intestinal epithelium. The genome of the probiotic Lactobacillus rhamnosus GG (LGG) carries a gene cluster encoding a putative L-fucose permease (fucP), L-fucose catabolic pathway (fucI, fucK, fucU, and fucA), and a transcriptional regulator (fucR). The metabolism of L-fucose in LGG results in 1,2-propanediol production, and their fucI and fucP mutants displayed a severe and mild growth defect on L-fucose, respectively. Transcriptional analysis revealed that the fuc genes are induced by L-fucose and subject to a strong carbon catabolite repression effect. This induction was triggered by FucR, which acted as a transcriptional activator necessary for growth on L-fucose. LGG utilized fucosyl-α1,3-N-acetylglucosamine and contrarily to other lactobacilli, the presence of fuc genes allowed this strain to use the L-fucose moiety. In fucI and fucR mutants, but not in fucP mutant, L-fucose was not metabolized and it was excreted to the medium during growth on fucosyl-α1,3-N-acetylglucosamine. The fuc genes were induced by this fucosyl-disaccharide in the wild type and the fucP mutant but not in a fucI mutant, showing that FucP does not participate in the regulation of fuc genes and that L-fucose metabolism is needed for FucR activation. The l-fucose operon characterized here constitutes a new example of the many factors found in LGG that allow this strain to adapt to the gastrointestinal conditions. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  17. Build Rocky Flats Environmental Technology site production prototype modular treatment system for stand alone core capability for residue unpack, sort, assay, repack

    International Nuclear Information System (INIS)

    Hildner, R.A.; Zygmunt, S.J.

    1997-01-01

    This document describes a portable and modular suit of equipment that upfront and near-term accomplishes a sorting process that documents and removes Rocky Flats Environmental Technology Site (RFETS) residue and waste from site inventory

  18. Cesium residue leachate migration in the tailings management area of a mine site : predicted vs. actual

    Energy Technology Data Exchange (ETDEWEB)

    Solylo, P.; Ramsey, D. [Wardrop Engineering, Winnipeg, MB (Canada). Mining and Minerals Section

    2009-07-01

    This paper reported on a study at a cesium products facility (CPF) that manufactures a non-toxic cesium-formate drilling fluid. The facility operates adjacent to a pollucite/tantalum/spodumene mine. The CPF was developed as a closed system, with the residue tailings slurry from the CPF process discharged to doublelined containment cells. Groundwater monitoring has shown that leachate has affected near-surface porewater quality within the tailings management area (TMA). Elevated concentrations of calcium, sulphate, strontium, cesium, and rubidium were used to identify the leachate. Porewater at the base of the tailings and in the overburden beneath the tailings has not been affected. A geochemical investigation was initiated to determine how the leachate behaves in the groundwater/tailings porewater system. Over the past 7 years of residue placement in the TMA, the footprint of the residue placement area has changed, making the comparison of predicted versus actual rate of leachate migration very subjective and difficult to quantify. Based solely on the analytical data, the source of the leachate is unknown, either from the original residue pile or the 2007 residue placement area. For purposes of long term residue management, an investigation of the geochemical behaviour of residue leachate in the groundwater/tailings system of the TMA is currently underway. 5 refs., 1 tab., 2 figs.

  19. Three-dimensional modeling of the Nevada Test Site and vicinity from teleseismic p-wave residuals

    International Nuclear Information System (INIS)

    Monfort, M.E.; Evans, J.R.

    1982-01-01

    A teleseismic P-wave travel-time residual study is described which reveals the regional compressional-velocity structure of southern Nevada and neighboring parts of California to a depth of 280 km. During 1980, 98 teleseismic events were recorded at 53 sites. P-wave residuals were calculated relative to a network-wide average residual for each event and are displayed on maps of the stations for each of four event-azimuth quadrants. Fluctuations in these map-patterns of residuals with approach azimuth combined with results of linear, three-dimensional inversions of some 2887 residuals indicate the following characteristics of the velocity structure of the southern Nevada region: (1) a low-velocity body exists in the upper crust 50 km northeast of Beatty, Nevada, near the Miocene Timber Mountain-Silent Canyon caldera complex. Another highly localized low-velocity anomaly occurs near the southwest corner of the Nevada Test Site (NTS). These two anomalies seem to be part of a low-velocity trough extending from Death Valley, California, to about 50 km north of NTS; (2) there is a high-velocity body in the mantle between 81 and 131 km deep centered about 10 km north of the edge of the Timber Mountain caldera; (3) a broad low-velocity body is delineated between 81 and 131 km deep centered about 30 km north of Las Vegas; (4) there is a monotonic increase in travel-time delays from west to east across the region, probably indicating an eastward decrease in velocity, and lower than average velocities in southeastern Nevada below 31 km; and (5) considerable complexity in three-dimensional velocity structure exists in this part of the southern Great Basin. Inversions of teleseismic P-wave travel-time residuals were also performed on data from 12 seismometers in the immediate vicinity of the NTS to make good use of the closer station spacing in that area

  20. Residues in the H+ Translocation Site Define the pKa for Sugar Binding to LacY†

    Science.gov (United States)

    Smirnova, Irina; Kasho, Vladimir; Sugihara, Junichi; Choe, Jun-Yong; Kaback, H. Ronald

    2009-01-01

    A remarkably high pKa of approximately 10.5 has been determined for sugar-binding affinity to the lactose permease of Escherichia coli (LacY), indicating that, under physiological conditions, substrate binds to fully protonated LacY. We have now systematically tested site-directed replacements for the residues involved in sugar binding, as well as H+ translocation and coupling, in order to determine which residues may be responsible for this alkaline pKa. Mutations in the sugar-binding site (Glu126, Trp151, Glu269) markedly decrease affinity for sugar but do not alter the pKa for binding. In contrast, replacements for residues involved in H+ translocation (Arg302, Tyr236, His322, Asp240, Glu325, Lys319) exhibit pKa values for sugar binding that are either shifted toward neutral pH or independent of pH. Values for the apparent dissociation constant for sugar binding (Kdapp) increase greatly for all mutants except neutral replacements for Glu325 or Lys319, which are characterized by remarkably high affinity sugar binding (i.e., low Kdapp) from pH 5.5 to pH 11. The pH dependence of the on- and off-rate constants for sugar binding measured directly by stopped-flow fluorometry implicates koff as a major factor for the affinity change at alkaline pH and confirms the effects of pH on Kdapp inferred from steady-state fluorometry. These results indicate that the high pKa for sugar binding by wild-type LacY cannot be ascribed to any single amino acid residue but appears to reside within a complex of residues involved in H+ translocation. There is structural evidence for water bound in this complex, and the water could be the site of protonation responsible for the pH dependence of sugar binding. PMID:19689129

  1. Characterization of an fungal l-fucokinase involved in Mortierella alpina GDP-l-fucose salvage pathway.

    Science.gov (United States)

    Wang, Hongchao; Zhang, Chen; Chen, Haiqin; Yang, Qin; Zhou, Xin; Gu, Zhennan; Zhang, Hao; Chen, Wei; Chen, Yong Q

    2016-08-01

    GDP-l-fucose functions as a biological donor for fucosyltransferases, which are required for the catalysis of l-fucose to various acceptor molecules including oligosaccharides, glycoproteins and glycolipids. Mortierella alpina is one of the highest lipid-producing fungi and can biosynthesis GDP-l-fucose in the de novo pathway. Analysis of the M. alpina genome suggests that there is a gene encoding l-fucokinase (FUK) for the conversion of fucose to l-fucose-1-phosphate in the GDP-l-fucose salvage pathway, which has never been found in fungi before. This gene was characterized to explore its role in GDP-l-fucose synthesis. The yield of GDP-l-fucose is relatively higher in lipid accumulation phase (0.096 mg per g cell) than that in cell multiplication phase (0.074 mg per g cell) of M. alpina Additionally, the transcript level of FUK is up regulated by nitrogen exhaustion when M. alpina starts to accumulate lipid, highlights the functional significance of FUK in the GDP-l-fucose biosynthesis in M. alpina Gene encoding FUK was expressed heterologously in Escherichia coli and the resulting protein was purified to homogeneity. The product of FUK reaction was analyzed by liquid chromatography and mass spectrometry. Kinetic parameters and other properties of FUK were investigated. Comparative analyses between the FUK protein and other homologous proteins were performed. To our knowledge, this study is the first to report a comprehensive characterization of FUK in a fungus. Mortierella alpina could be used as an alternative source for the production of GDP-l-fucose. © The Author 2016. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  2. Identification of residues involved in nucleotidyltransferase activity of JHP933 from helicobacter pyloriby site-directed mutagenesis

    Directory of Open Access Journals (Sweden)

    Ye Xianren

    2016-01-01

    Full Text Available Helicobacter pylori is a well-known bacterial pathogen involved in the development of peptic ulcer, gastric adenocarcinoma and other forms of gastric cancer. Evidence has suggested that certain strain-specific genes in the plasticity region may play key roles in the pathogenesis of H. pylori-associated gastroduodenal diseases. Therefore there is considerable interest in the strain-specific genes located in the plasticity regions of H. pylori. JHP933 is encoded by the gene in the plasticity region of H. pylori strain J99. Recently, the crystal structure of JHP933 has confirmed it as a nucleotidyltransferase (NTase superfamily protein and a putative active site has been proposed. However, no evidence from direct functional assay has been presented to confirm the active site and little is known about the functional mechanism of JHP933. Here, through superimposition with Cid1/NTP complex structures, we modelled the complex structures of JHP933 with different NTPs. Based on the models and using rational site-directed mutagenesis combined with enzymatic activity assays, we confirm the active site and identify several residues important for the nucleotidyl transferring function of JHP933. Furthermore, mutations of these active site residues result in the abolishment of the nucleotidyltransferase activity of JHP933. This work provides preliminary insight into the molecular mechanism underlying the pathophysiological role in H. pylori infection of JHP933 as a novel NTase superfamily protein.

  3. Derivation of guidelines for uranium residual radioactive material in soil at the B ampersand T Metals Company site, Columbus, Ohio

    International Nuclear Information System (INIS)

    Kamboj, S.; Nimmagadda, Mm.; Yu, C.

    1996-01-01

    Guidelines for uranium residual radioactive material in soil were derived for the B ampersand T Metals Company site in Columbus, Ohio. This site has been identified for remedial action under the US Department of Energy's (DOE's) Formerly Utilized Sites Remedial Action Program (FUSRAP). Single-nuclide and total-uranium guidelines were derived on the basis of the requirement that following remedial action, the 50-year committed effective dose equivalent to a hypothetical individual living or working in the immediate vicinity of the site should not exceed a dose constraint of 30 mrem/yr for the current use and likely future use scenarios or a dose limit of 100 n-mrem/yr for less likely future use scenarios. The DOE residual radioactive material guideline computer code, RESRAD, was used in this evaluation. RESRAD implements the methodology described in the DOE manual for establishing residual radioactive material guidelines. Three scenarios were considered; each assumed that for a period of 1,000 years following remedial action, the site would be used without radiological restrictions. The three scenarios varied with regard to the type of site use, time spent at the site by the exposed individual, and sources of food and water consumed. The evaluations indicate that the dose constraint of 30 mrem/yr would not be exceeded for uranium (including uranium-234, uranium-235, and uranium-238) within 1,000 years, provided that the soil concentration of total uranium (uranium-234, uranium-235, and uranium-238) at the B ampersand T Metals site did not exceed 1, I 00 pCi/g for Scenario A (industrial worker, current use) or 300 pCi/g for Scenario B (resident with municipal water supply, a likely future use). The dose limit of 100 mrem/yr would not be exceeded at the site if the total uranium concentration of the soil did not exceed 880 pCi/g for Scenario C (resident with an on-site water well, a plausible but unlikely future use)

  4. Hanford Site Tank 241-C-108 Residual Waste Contaminant Release Models and Supporting Data

    Energy Technology Data Exchange (ETDEWEB)

    Cantrell, Kirk J.; Krupka, Kenneth M.; Geiszler, Keith N.; Arey, Bruce W.; Schaef, Herbert T.

    2010-06-18

    This report presents the results of laboratory characterization, testing, and analysis for a composite sample (designated 20578) of residual waste collected from single-shell tank C-108 during the waste retrieval process after modified sluicing. These studies were completed to characterize concentration and form of contaminant of interest in the residual waste; assess the leachability of contaminants from the solids; and develop release models for contaminants of interest. Because modified sluicing did not achieve 99% removal of the waste, it is expected that additional retrieval processing will take place. As a result, the sample analyzed here is not expected to represent final retrieval sample.

  5. A novel bicistronic gene design couples stable cell line selection with a fucose switch in a designer CHO host to produce native and afucosylated glycoform antibodies.

    Science.gov (United States)

    Roy, Gargi; Martin, Tom; Barnes, Arnita; Wang, Jihong; Jimenez, Rod Brian; Rice, Megan; Li, Lina; Feng, Hui; Zhang, Shu; Chaerkady, Raghothama; Wu, Herren; Marelli, Marcello; Hatton, Diane; Zhu, Jie; Bowen, Michael A

    2018-04-01

    The conserved glycosylation site Asn 297 of a monoclonal antibody (mAb) can be decorated with a variety of sugars that can alter mAb pharmacokinetics and recruitment of effector proteins. Antibodies lacking the core fucose at Asn 297 (afucosylated mAbs) show enhanced antibody-dependent cell-mediated cytotoxicity (ADCC) and increased efficacy. Here, we describe the development of a robust platform for the manufacture of afucosylated therapeutic mAbs by engineering a Chinese hamster ovary (CHO) host cell line to co-express a mAb with GDP-6-deoxy-D-lyxo-4-hexulose reductase (RMD), a prokaryotic enzyme that deflects an intermediate in the de novo synthesis of fucose to a dead-end product, resulting in the production of afucosylated mAb (GlymaxX™ Technology, ProBioGen). Expression of the mAb and RMD genes was coordinated by co-transfection of separate mAb and RMD vectors or use of an internal ribosome entry site (IRES) element to link the translation of RMD with either the glutamine synthase selection marker or the mAb light chain. The GS-IRES-RMD vector format was more suitable for the rapid generation of high yielding cell lines, secreting afucosylated mAb with titers exceeding 6.0 g/L. These cell lines maintained production of afucosylated mAb over 60 generations, ensuring their suitability for use in large-scale manufacturing. The afucosylated mAbs purified from these RMD-engineered cell lines showed increased binding in a CD16 cellular assay, demonstrating enhancement of ADCC compared to fucosylated control mAb. Furthermore, the afucosylation in these mAbs could be controlled by simple addition of L-fucose in the culture medium, thereby allowing the use of a single cell line for production of the same mAb in fucosylated and afucosylated formats for multiple therapeutic indications.

  6. Comparison of long-term stability of containment systems for residues and wastes contaminated with naturally occurring radionuclides at an arid site and two humid sites

    International Nuclear Information System (INIS)

    Winters, M.; Merry-Libby, P.; Hinchman, R.

    1985-01-01

    The long-term stability of near-surface containment systems designed for the management of radioactive wastes and residues contaminated with naturally occurring radionuclides are compared at the three different sites. The containment designs are: (1) a diked 8.9-m high mound, including a 3.2-m layered cap at a site (humid) near Lewiston, New York, (2) a 6.8-m-high mound, including a similar 3.2-m cap at a site (humid) near Oak Ridge, Tennessee, and (3) 4.8-m deep trenches with 3.0-m backfilled caps at a site (arid) near Hanford, Washington. Geological, hydrological, and biological factors affecting the long-term (1000-year) integrity of the containment systems at each site are examined, including: erosion, flooding, drought, wildfire, slope and cover failure, plant root penetration, burrowing animals, other soil-forming processes, and land-use changes. For the containment designs evaluated, releases of radon-222 at the arid site are predicted to be several orders of magnitude higher than at the two humid sites - upon initial burial and at 1000 years (after severe erosion). Transfer of wastes containing naturally occurring radionuclides from a humid to an arid environment offers little or no advantage relative to long-term stability of the containment system and has a definite disadvantage in terms of gaseous radioactive releases. 26 references, 3 figures, 4 tables

  7. Evaluation of serum sialic acid, fucose levels and their ratio in oral squamous cell carcinoma.

    Science.gov (United States)

    Chinnannavar, Sangamesh Ningappa; Ashok, Lingappa; Vidya, Kodige Chandrashekhar; Setty, Sunil Mysore Kantharaja; Narasimha, Guru Eraiah; Garg, Ranjana

    2015-01-01

    Detection of cancer at the early stage is of utmost importance to decrease the morbidity and mortality of the disease. Apart from the conventional biopsy, minimally invasive methods like serum evaluation are used for screening large populations. Thus, this study aimed to estimate serum levels of sialic acid and fucose and their ratio in oral cancer patients and in healthy control group to evaluate their role in diagnosis. Serum samples were collected from 52 healthy controls (group I) and 52 squamous cell carcinoma patients (group II). Estimation of serum levels of sialic acid and fucose and their ratio was performed. This was correlated histopathologically with the grades of carcinoma. Statistical analysis was done by using analysis of variance (ANOVA) test and unpaired "t" test. Results showed that serum levels of sialic acid and fucose were significantly higher in oral cancer patients compared to normal healthy controls (P ratio was significantly lower in cancer patients than in normal controls (P ratio showed decreasing trend from controls to malignant group. The ratio of sialic acid to fucose can be a useful diagnostic aid for oral cancer patients.

  8. GDP-L-fucose is required for boundary definition in plants.

    Science.gov (United States)

    Gonçalves, Beatriz; Maugarny-Calès, Aude; Adroher, Bernard; Cortizo, Millán; Borrega, Nero; Blein, Thomas; Hasson, Alice; Gineau, Emilie; Mouille, Grégory; Laufs, Patrick; Arnaud, Nicolas

    2017-12-16

    The CUP-SHAPED COTYLEDON (CUC) transcription factors control plant boundary formation, thus allowing the emergence of novel growth axes. While the developmental roles of the CUC genes in different organs and across species are well characterized, upstream and downstream events that contribute to their function are still poorly understood. To identify new players in this network, we performed a suppressor screen of CUC2g-m4, a line overexpressing CUC2 that has highly serrated leaves. We identified a mutation that simplifies leaf shape and affects MURUS1 (MUR1), which is responsible for GDP-L-fucose production. Using detailed morphometric analysis, we show that GDP-L-fucose has an essential role in leaf shape acquisition by sustaining differential growth at the leaf margins. Accordingly, reduced CUC2 expression levels are observed in mur1 leaves. Furthermore, genetic analyses reveal a conserved role for GDP-L-fucose in different developmental contexts where it contributes to organ separation in the same pathway as CUC2. Taken together, our results reveal that GDP-L-fucose is necessary for proper establishment of boundary domains in various developmental contexts. © The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  9. Trophic Interactions of Infant Bifidobacteria and Eubacterium hallii during L-Fucose and Fucosyllactose Degradation.

    Science.gov (United States)

    Schwab, Clarissa; Ruscheweyh, Hans-Joachim; Bunesova, Vera; Pham, Van Thanh; Beerenwinkel, Niko; Lacroix, Christophe

    2017-01-01

    Fucosyllactoses (2'- or 3'-FL) account for up to 20% of human milk oligosaccharides (HMOs). Infant bifidobacteria, such as Bifidobacterium longum subsp. infantis , utilize the lactose moiety to form lactate and acetate, and metabolize L-fucose to 1,2-propanediol (1,2-PD). Eubacterium hallii is a common member of the adult gut microbiota that can produce butyrate from lactate and acetate, and convert 1,2-PD to propionate. Recently, a Swiss cohort study identified E. hallii as one of the first butyrate producers in the infant gut. However, the global prevalence of E. hallii and its role in utilization of HMO degradation intermediates remains unexplored. Fecal 16S rRNA gene libraries ( n = 857) of humans of all age groups from Venezuela, Malawi, Switzerland, and the USA were screened for the occurrence of E. hallii . Single and co-culture experiments of B. longum subsp. infantis and E. hallii were conducted in modified YCFA containing acetate and glucose, L-fucose, or FL. Bifidobacterium spp. ( n = 56) of different origin were screened for the ability to metabolize L-fucose. Relative abundance of E. hallii was low (10 -5 -10 -3 %) during the first months but increased and reached adult levels (0.01-10%) at 5-10 years of age in all four populations. In single culture, B. longum subsp. infantis grew in the presence of all three carbohydrates while E. hallii was metabolically active only with glucose. In co-culture E. hallii also grew with L-fucose or FL. In co-cultures grown with glucose, acetate, and glucose were consumed and nearly equimolar proportions of formate and butyrate were formed. B. longum subsp. infantis used L-fucose and produced 1,2-PD, acetate and formate in a ratio of 1:1:1, while 1,2-PD was used by E. hallii to form propionate. E. hallii consumed acetate, lactate and 1,2-PD released by B. longum subsp. infantis from FL, and produced butyrate, propionate, and formate. Beside B. longum subsp. infantis, Bifidobacterium breve , and a strain of B. longum

  10. Important Determinants for Fucoidan Bioactivity: A Critical Review of Structure-Function Relations and Extraction Methods for Fucose-Containing Sulfated Polysaccharides from Brown Seaweeds

    Directory of Open Access Journals (Sweden)

    Anne S. Meyer

    2011-10-01

    Full Text Available Seaweeds—or marine macroalgae—notably brown seaweeds in the class Phaeophyceae, contain fucoidan. Fucoidan designates a group of certain fucose-containing sulfated polysaccharides (FCSPs that have a backbone built of (1→3-linked α-l-fucopyranosyl or of alternating (1→3- and (1→4-linked α-l-fucopyranosyl residues, but also include sulfated galactofucans with backbones built of (1→6-β-d-galacto- and/or (1→2-β-d-mannopyranosyl units with fucose or fuco-oligosaccharide branching, and/or glucuronic acid, xylose or glucose substitutions. These FCSPs offer several potentially beneficial bioactive functions for humans. The bioactive properties may vary depending on the source of seaweed, the compositional and structural traits, the content (charge density, distribution, and bonding of the sulfate substitutions, and the purity of the FCSP product. The preservation of the structural integrity of the FCSP molecules essentially depends on the extraction methodology which has a crucial, but partly overlooked, significance for obtaining the relevant structural features required for specific biological activities and for elucidating structure-function relations. The aim of this review is to provide information on the most recent developments in the chemistry of fucoidan/FCSPs emphasizing the significance of different extraction techniques for the structural composition and biological activity with particular focus on sulfate groups.

  11. Engineering feasibility analysis for in-situ stabilization of Burrell Township site residues

    International Nuclear Information System (INIS)

    1982-11-01

    The Burrell Township site, located in western Pennsylvania, received approximately 11,600 tons of radioactively-contaminated material in late 1956 and early 1957 from the Vitro Manufacturing Company's operations in Canonsburg, Pennsylvania. WESTON was requested to conduct an engineering study to determine the feasibility of stabilizing the site in accordance with the US Environmental Protection Agency's (EPA) interim and proposed standards (45 FR 27366--27368, April 22, 1980, and 46 FR 2556--2563, January 9, 1981). The scope of this study is limited to those alternatives that can be implemented on the site and will not require removal and offsite disposal of radioactively-contaminated material. Four alternatives for control of the radioactive material at the Burrell site were considered and evaluated, as follows: 1. Site stabilization and closure. 2. Site control and containment. 3. Waste excavation and encapsulation. 4. Waste excavation, incineration, and encapsulation. 2 refs., 32 figs., 12 tabs

  12. A tiered analytical protocol for the characterization of heavy oil residues at petroleum-contaminated hazardous waste sites

    International Nuclear Information System (INIS)

    Pollard, S.J.T.; Kenefick, S.L.; Hrudey, S.E.; Fuhr, B.J.; Holloway, L.R.; Rawluk, M.

    1994-01-01

    The analysis of hydrocarbon-contaminated soils from abandoned refinery sites in Alberta, Canada is used to illustrate a tiered analytical approach to the characterization of complex hydrocarbon wastes. Soil extracts isolated from heavy oil- and creosote-contaminated sites were characterized by thin layer chromatography with flame ionization detection (TLC-FID), ultraviolet fluorescence, simulated distillation (GC-SIMDIS) and chemical ionization GC-MS analysis. The combined screening and detailed analytical methods provided information essential to remedial technology selection including the extent of contamination, the class composition of soil extracts, the distillation profile of component classes and the distribution of individual class components within various waste fractions. Residual contamination was characteristic of heavy, degraded oils, consistent with documented site operations and length of hydrocarbon exposure at the soil surface

  13. Metabolic engineering of Corynebacterium glutamicum to produce GDP-L-fucose from glucose and mannose.

    Science.gov (United States)

    Chin, Young-Wook; Park, Jin-Byung; Park, Yong-Cheol; Kim, Kyoung Heon; Seo, Jin-Ho

    2013-06-01

    Wild-type Corynebacterium glutamicum was metabolically engineered to convert glucose and mannose into guanosine 5'-diphosphate (GDP)-L-fucose, a precursor of fucosyl-oligosaccharides, which are involved in various biological and pathological functions. This was done by introducing the gmd and wcaG genes of Escherichia coli encoding GDP-D-mannose-4,6-dehydratase and GDP-4-keto-6-deoxy-D-mannose-3,5-epimerase-4-reductase, respectively, which are known as key enzymes in the production of GDP-L-fucose from GDP-D-mannose. Coexpression of the genes allowed the recombinant C. glutamicum cells to produce GDP-L-fucose in a minimal medium containing glucose and mannose as carbon sources. The specific product formation rate was much higher during growth on mannose than on glucose. In addition, the specific product formation rate was further increased by coexpressing the endogenous phosphomanno-mutase gene (manB) and GTP-mannose-1-phosphate guanylyl-transferase gene (manC), which are involved in the conversion of mannose-6-phosphate into GDP-D-mannose. However, the overexpression of manA encoding mannose-6-phosphate isomerase, catalyzing interconversion of mannose-6-phosphate and fructose-6-phosphate showed a negative effect on formation of the target product. Overall, coexpression of gmd, wcaG, manB and manC in C. glutamicum enabled production of GDP-L-fucose at the specific rate of 0.11 mg g cell(-1) h(-1). The specific GDP-L-fucose content reached 5.5 mg g cell(-1), which is a 2.4-fold higher than that of the recombinant E. coli overexpressing gmd, wcaG, manB and manC under comparable conditions. Well-established metabolic engineering tools may permit optimization of the carbon and cofactor metabolisms of C. glutamicum to further improve their production capacity.

  14. Biochemical characterization of GDP-L-fucose de novo synthesis pathway in fungus Mortierella alpina

    International Nuclear Information System (INIS)

    Ren, Yan; Perepelov, Andrei V.; Wang, Haiyan; Zhang, Hao; Knirel, Yuriy A.; Wang, Lei; Chen, Wei

    2010-01-01

    Mortierella alpina is a filamentous fungus commonly found in soil, which is able to produce large amount of polyunsaturated fatty acids. L-Fucose is an important sugar found in a diverse range of organisms, playing a variety of biological roles. In this study, we characterized the de novo biosynthetic pathway of GDP-L-fucose (the nucleotide-activated form of L-fucose) in M. alpina. Genes encoding GDP-D-mannose 4,6-dehydratase (GMD) and GDP-keto-6-deoxymannose 3,5-epimerase/4-reductase (GMER) were expressed heterologously in Escherichia coli. The recombinant enzymes were produced as His-tagged fusion proteins. Conversion of GDP-mannose to GDP-4-keto-6-deoxy mannose by GMD and GDP-4-keto-6-deoxy mannose to GDP-L-fucose by GMER were analyzed by capillary electrophoresis, electro-spray ionization-mass spectrometry, and nuclear magnetic resonance spectroscopy. The k m values of GMD for GDP-mannose and GMER for GDP-4-keto-6-deoxy mannose were determined to be 0.77 mM and 1.047 mM, respectively. Both NADH and NADPH may be used by GMER as the coenzyme. The optimum temperature and pH were determined to be 37 o C and pH 9.0 (GMD) or pH 7.0 (GMER). Divalent cations are not required for GMD and GMER activity, and the activities of both enzymes may be enhanced by DTT. To our knowledge this is the first report on the characterization of GDP-L-fucose biosynthetic pathway in fungi.

  15. Sulfation pattern of fucose branches affects the anti-hyperlipidemic activities of fucosylated chondroitin sulfate.

    Science.gov (United States)

    Wu, Nian; Zhang, Yu; Ye, Xingqian; Hu, Yaqin; Ding, Tian; Chen, Shiguo

    2016-08-20

    Fucosylated chondroitin sulfates (fCSs) are glycosaminoglycans extracted from sea cucumbers, consisting of chondroitin sulfate E (CSE) backbones and sulfated fucose branches. The biological properties of fCSs could be affected by the sulfation pattern of their fucose branches. In the present study, two fCSs were isolated from sea cucumbers Isostichopus badionotus (fCS-Ib) and Pearsonothuria graeffei (fCS-Pg). Their monosaccharide compositions of glucuronic acid (GlcA), N-acetylgalactosamine (GalNAc), fucose (Fuc) and sulfate were at similar molar ratio with 1.0/0.7/0.9/3.1 for fCS-Ib and 1.0/0.8/1.5/2.6 for fCS-Pg. The two fCSs have different sulfation patterns on their fucose branches, fCS-Pg with 3,4-O-disulfation while fCS-Ib with 2,4-O-disulfation. Their antihyperlipidemic effects were compared using a high-fat high-fructose diet (HFFD)-fed C57BL/6J mice model. Both fCS-Ib and fCS-Pg had significant effects on lipid profile improvement, liver protection, blood glucose diminution and hepatic glycogen synthesis. Specifically, fCS-Pg with 3,4-O-disulfation fucose branches was more effective in reduction of blood cholesterol (TC), low density lipoprotein (LDL) and atherogenic index (AI). Our results indicate that both fCSs, especially fCS-Pg, could be used as a potential anti-hyperlipidemic drug. Copyright © 2016. Published by Elsevier Ltd.

  16. Physical stability comparisons of IgG1-Fc variants: effects of N-glycosylation site occupancy and Asp/Gln residues at site Asn 297.

    Science.gov (United States)

    Alsenaidy, Mohammad A; Okbazghi, Solomon Z; Kim, Jae Hyun; Joshi, Sangeeta B; Middaugh, C Russell; Tolbert, Thomas J; Volkin, David B

    2014-06-01

    The structural integrity and conformational stability of various IgG1-Fc proteins produced from the yeast Pichia pastoris with different glycosylation site occupancy (di-, mono-, and nonglycosylated) were determined. In addition, the physical stability profiles of three different forms of nonglycosylated Fc molecules (varying amino-acid residues at site 297 in the CH 2 domain due to the point mutations and enzymatic digestion of the Fc glycoforms) were also examined. The physical stability of these IgG1-Fc glycoproteins was examined as a function of pH and temperature by high-throughput biophysical analysis using multiple techniques combined with data visualization tools (three index empirical phase diagrams and radar charts). Across the pH range of 4.0-6.0, the di- and monoglycosylated forms of the IgG1-Fc showed the highest and lowest levels of physical stability, respectively, with the nonglycosylated forms showing intermediate stability depending on solution pH. In the aglycosylated Fc proteins, the introduction of Asp (D) residues at site 297 (QQ vs. DN vs. DD forms) resulted in more subtle changes in structural integrity and physical stability depending on solution pH. The utility of evaluating the conformational stability profile differences between the various IgG1-Fc glycoproteins is discussed in the context of analytical comparability studies. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

  17. Site directed mutagenesis of amino acid residues at the active site of mouse aldehyde oxidase AOX1.

    Directory of Open Access Journals (Sweden)

    Silvia Schumann

    Full Text Available Mouse aldehyde oxidase (mAOX1 forms a homodimer and belongs to the xanthine oxidase family of molybdoenzymes which are characterized by an essential equatorial sulfur ligand coordinated to the molybdenum atom. In general, mammalian AOs are characterized by broad substrate specificity and an yet obscure physiological function. To define the physiological substrates and the enzymatic characteristics of mAOX1, we established a system for the heterologous expression of the enzyme in Escherichia coli. The recombinant protein showed spectral features and a range of substrate specificity similar to the native protein purified from mouse liver. The EPR data of recombinant mAOX1 were similar to those of AO from rabbit liver, but differed from the homologous xanthine oxidoreductase enzymes. Site-directed mutagenesis of amino acids Val806, Met884 and Glu1265 at the active site resulted in a drastic decrease in the oxidation of aldehydes with no increase in the oxidation of purine substrates. The double mutant V806E/M884R and the single mutant E1265Q were catalytically inactive enzymes regardless of the aldehyde or purine substrates tested. Our results show that only Glu1265 is essential for the catalytic activity by initiating the base-catalyzed mechanism of substrate oxidation. In addition, it is concluded that the substrate specificity of molybdo-flavoenzymes is more complex and not only defined by the three characterized amino acids in the active site.

  18. Cys-X scanning for expansion of active-site residues and modulation of catalytic functions in a glutathione transferase.

    Science.gov (United States)

    Norrgård, Malena A; Hellman, Ulf; Mannervik, Bengt

    2011-05-13

    We propose Cys-X scanning as a semisynthetic approach to engineer the functional properties of recombinant proteins. As in the case of Ala scanning, key residues in the primary structure are identified, and one of them is replaced by Cys via site-directed mutagenesis. The thiol of the residue introduced is subsequently modified by alternative chemical reagents to yield diverse Cys-X mutants of the protein. This chemical approach is orthogonal to Ala or Cys scanning and allows the expansion of the repertoire of amino acid side chains far beyond those present in natural proteins. In its present application, we have introduced Cys-X residues in human glutathione transferase (GST) M2-2, replacing Met-212 in the substrate-binding site. To achieve selectivity of the modifications, the Cys residues in the wild-type enzyme were replaced by Ala. A suite of simple substitutions resulted in a set of homologous Met derivatives ranging from normethionine to S-heptyl-cysteine. The chemical modifications were validated by HPLC and mass spectrometry. The derivatized mutant enzymes were assayed with alternative GST substrates representing diverse chemical reactions: aromatic substitution, epoxide opening, transnitrosylation, and addition to an ortho-quinone. The Cys substitutions had different effects on the alternative substrates and differentially enhanced or suppressed catalytic activities depending on both the Cys-X substitution and the substrate assayed. As a consequence, the enzyme specificity profile could be changed among the alternative substrates. The procedure lends itself to large-scale production of Cys-X modified protein variants.

  19. Criteria and measurement techniques applicable to residual radioactivity on a decommissioned reactor site

    International Nuclear Information System (INIS)

    Woollam, P.B.

    1988-12-01

    This document summarises the radiological criteria which might be developed to cover the release of a partly decommissioned nuclear reactor site, then looks at the techniques available by which the site could be monitored to assure compliance with these criteria. In particular, the implications of existing levels of radioactive contamination resulting from airburst nuclear weapons tests and the Chernobyl accident are discussed. (author)

  20. Characterization of Solids in Residual Wastes from Single-Shell Tanks at the Hanford Site, Washington, USA - 9277

    International Nuclear Information System (INIS)

    Krupka, Kenneth M.; Cantrell, Kirk J.; Schaef, Herbert T.; Arey, Bruce W.; Heald, Steve M.; Deutsch, William J.; Lindberg, Michael J.

    2009-01-01

    Solid-phase characterization methods have been used in an ongoing study of residual wastes (i.e., waste remaining after final retrieval operations) from the underground single-shell storage tanks 241-C-103, 241-C-106, 241-C-202, 241-C-203, and 241-S-112 at the U.S. Department of Energy's Hanford Site in Washington State. The results of studies completed to date show significant variability in the compositions of those residual wastes and the compositions, morphologies, and crystallinities of the individual phases that make up these wastes. These differences undoubtedly result from the various waste types stored and transferred in and out each tank and the sluicing and retrieval operations used for waste retrieval. Our studies indicate that these residual wastes are chemically-complex assemblages of crystalline and amorphous solids that contain contaminants as discrete phases and/or co-precipitated within oxide phases. Depending on the specific tank, various solids (e.g., gibbsite; boehmite; dawsonite; cancrinite; Fe oxides such as hematite, goethite, and maghemite; rhodochrosite; lindbergite; whewellite; nitratine; and numerous amorphous or poorly crystalline phases) have been identified by X-ray diffraction and scanning electron microscopy/energy dispersive X-ray spectroscopy in residual wastes studied to date. Our studies also show that contact of residual wastes with Ca(OH)2- and CaCO3-saturated aqueous solutions, which were used as surrogates for the compositions of pore-fluid leachants derived from young and aged cements respectively, may alter the compositions of solid phases present in the contacted wastes. Fe oxides/hydroxides have been identified in all residual wastes studied to date. They occur in these wastes as discrete particles, particles intergrown within a matrix of other phases, and surface coatings on other particles or particle aggregates. These Fe oxides/hydroxides typically contain trace concentrations of other transition metals, such Cr, Mn

  1. Understanding Which Residues of the Active Site and Loop Structure of a Tyrosine Aminomutase Define Its Mutase and Lyase Activities.

    Science.gov (United States)

    Attanayake, Gayanthi; Walter, Tyler; Walker, Kevin D

    2018-05-30

    Site-directed mutations and substrate analogues were used to gain insights into the branch-point reaction of the 3,5-dihydro-5-methylidene-4 H-imidazol-4-one (MIO)-tyrosine aminomutase from Oryza sativa ( OsTAM). Exchanging the active residues of OsTAM (Y125C/N446K) for those in a phenylalanine aminomutase TcPAM altered its substrate specificity from tyrosine to phenylalanine. The aminomutase mechanism of OsTAM surprisingly changed almost exclusively to that of an ammonia lyase making cinnamic acid (>95%) over β-phenylalanine [Walter, T., et al. (2016) Biochemistry 55, 3497-3503]. We hypothesized that the missing electronics or sterics on the aryl ring of the phenylalanine substrate, compared with the sizable electron-donating hydroxyl of the natural tyrosine substrate, influenced the unexpected lyase reactivity of the OsTAM mutant. The double mutant was incubated with 16 α-phenylalanine substituent analogues of varying electronic strengths and sterics. The mutant converted each analogue principally to its acrylate with ∼50% conversion of the p-Br substrate, making only a small amount of the β-amino acid. The inner loop structure over the entrance to the active site was also mutated to assess how the lyase and mutase activities are affected. An OsTAM loop mutant, matching the loop residues of TcPAM, still chiefly made >95% of the acrylate from each substrate. A combined active site:loop mutant was most reactive but remained a lyase, making 10-fold more acrylates than other mutants did. While mutations within the active site changed the substrate specificity of OsTAM, continued exploration is needed to fully understand the interplay among the inner loop, the substrate, and the active site in defining the mutase and lyase activities.

  2. Feasibility study for remedial action for the Quarry Residuals Operable Unit at the Weldon Spring Site, Weldon Spring, Missouri

    International Nuclear Information System (INIS)

    1998-03-01

    The U.S. Department of Energy (DOE) is conducting cleanup activities at the Weldon Spring site, which is located in St. Charles County, Missouri, about 48 km (30 mi) west of St. Louis (Figure 1.1). Cleanup of the Weldon Spring site consists of several integrated components. The quarry residuals operable unit (QROU) is one of four operable units being evaluated. In accordance with the Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA), as amended, a remedial investigation/feasibility study (RI/FS) is being conducted to evaluate conditions and potential responses for the following areas and/or media that constitute the QROU: (1) the residual material (soil and sediment) remaining at the Weldon Spring quarry after removal of the bulk waste (about 11 million L [3 million gal] of uranium-contaminated ponded water was also addressed previous to bulk waste removal); (2) other media located in the surrounding vicinity of the quarry, including adjacent soil, surface water, and sediment in Femme Osage Slough and several creeks; and (3) quarry groundwater located primarily north of Femme Osage Slough. Potential impacts to the St. Charles County well field downgradient of the quarry area are also being addressed as part of QROU RI/FS evaluations. For remedial action sites, it is DOE policy to integrate values associated with the National Environmental Policy Act (NEPA) into the CERCLA decision-making process. The analyses contained herein address NEPA values as appropriate to the actions being considered for the QROU. A work plan summarizing initial site conditions and providing conceptual site hydrogeological and exposure models was published in January 1994. The RI and baseline risk assessment (BRA) reports have been completed. The RI discusses in detail the nature and extent and the fate and transport of contamination at the quarry area

  3. Feasibility study for remedial action for the Quarry Residuals Operable Unit at the Weldon Spring Site, Weldon Spring, Missouri

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1998-03-01

    The U.S. Department of Energy (DOE) is conducting cleanup activities at the Weldon Spring site, which is located in St. Charles County, Missouri, about 48 km (30 mi) west of St. Louis (Figure 1.1). Cleanup of the Weldon Spring site consists of several integrated components. The quarry residuals operable unit (QROU) is one of four operable units being evaluated. In accordance with the Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA), as amended, a remedial investigation/feasibility study (RI/FS) is being conducted to evaluate conditions and potential responses for the following areas and/or media that constitute the QROU: (1) the residual material (soil and sediment) remaining at the Weldon Spring quarry after removal of the bulk waste (about 11 million L [3 million gal] of uranium-contaminated ponded water was also addressed previous to bulk waste removal); (2) other media located in the surrounding vicinity of the quarry, including adjacent soil, surface water, and sediment in Femme Osage Slough and several creeks; and (3) quarry groundwater located primarily north of Femme Osage Slough. Potential impacts to the St. Charles County well field downgradient of the quarry area are also being addressed as part of QROU RI/FS evaluations. For remedial action sites, it is DOE policy to integrate values associated with the National Environmental Policy Act (NEPA) into the CERCLA decision-making process. The analyses contained herein address NEPA values as appropriate to the actions being considered for the QROU. A work plan summarizing initial site conditions and providing conceptual site hydrogeological and exposure models was published in January 1994. The RI and baseline risk assessment (BRA) reports have been completed. The RI discusses in detail the nature and extent and the fate and transport of contamination at the quarry area.

  4. Residual radioactivity guidelines for the heavy water components test reactor at the Savannah River Site

    International Nuclear Information System (INIS)

    Owen, M.B. Smith, R.; McNeil, J.

    1997-04-01

    Guidelines were developed for acceptable levels of residual radioactivity in the Heavy Water Components Test Reactor (HWCTR) facility at the conclusion of its decommissioning. Using source terms developed from data generated in a detailed characterization study, the RESRAD and RASRAD-BUILD computer codes were used to calculate derived concentration guideline levels (DCGLs) for the radionuclides that will remain in the facility. The calculated DCGLs, when compared to existing concentrations of radionuclides measured during a 1996 characterization program, indicate that no decontamination of concrete surfaces will be necessary. Also, based on the results of the calculations, activated concrete in the reactor biological shield does not have to be removed, and imbedded radioactive piping in the facility can remain in place. Viewed in another way, the results of the calculations showed that the present inventory of residual radioactivity in the facility (not including that associated with the reactor vessel and steam generators) would produce less than one millirem per year above background to a hypothetical individual on the property. The residual radioactivity is estimated to be approximately 0.04 percent of the total inventory in the facility as of March, 1997. According to the results, the only radionuclides that would produce greater than 0.0.1-millirem per year are Am-241 (0.013 mrem/yr at 300 years), C-14 (0.022 mrem/yr at 1000 years) and U-238 (0.034 mrem/yr at 6000 years). Human exposure would occur only through the groundwater pathways, that is, from water drawn from, a well on the property. The maximum exposure would be approximately one percent of the 4 millirem per year ground water exposure limit established by the U.S. Environmental Protection Agency. 11 refs., 13 figs., 15 tabs

  5. Radioactive residues at nuclear sites in the United States of America

    International Nuclear Information System (INIS)

    Fiore, J.; Rampertaap, A.; Greeves, J.; MacKinney, J.; Raguso, M.; Selstrom, J.

    2000-01-01

    The United States of America has a large number of sites where radioactive materials have been mined, processed, produced, or used. Upon completion of activities at these sites, some will be cleaned up completely and released for unrestricted future use. However, at other sites, contamination will remain on the site following cleanup, requiring long term institutional controls. Depending on the purpose or the activity conducted at a specific site, these locations are generally under the jurisdiction of a federal agency: the United States Department of Energy, the United States Nuclear Regulatory Commission, the United States Environmental Protection Agency, or the United States Department of Defense. However, under certain conditions, the Nuclear Regulatory Commission relinquishes regulatory authority to individual states to regulate the commercial uses of radioactive materials (except for nuclear reactors), so some locations with radioactive materials are under the jurisdiction of individual states. Other sites with naturally occurring radioactive contamination may also be controlled by individual states. One or two sites under the jurisdiction of each agency are discussed where radioactive materials either existed in the past or exist today, and the source of contamination, the anticipated end state, and the process by which the responsible agency did or will conduct cleanup and site closure are described. Several issues are reviewed that must be addressed in order to design and implement remediations that will ensure long term protection for the environment and future inhabitants. The role of citizens in environmental cleanups is examined, and how institutional controls may be applied to ensure long term protection of remediations that leave some contaminants in place. (author)

  6. Synthesis and evaluation of radioactive and fluorescent residualizing labels for identifying sites of plasma protein catabolism

    International Nuclear Information System (INIS)

    Maxwell, J.L.; Baynes, J.W.; Thorpe, S.R.

    1986-01-01

    Inulin and lactose were each coupled to tyramine by reductive amination with NaBH 3 CN and the tyramine then labeled with 125 I. Dilactitol- 125 I-tyramine (DLT) and inulin- 125 I-tyramine (InTn) were coupled by reductive amination and cyanuric chloride, respectively, to asialofetuin (ASF), fetuin and rat serum albumin (RSA). Attachment of either label had no effect on the circulating half-lives of the proteins. Radioactivity from labeled ASF was recovered in rat liver (> 90%) by 1 h post-injection and remained in liver with half-lives of 2 and 6 days, respectively, for the DLT and InTn labels. Whole body recoveries of radioactivity from DLT- and InTn labels. Whole body recoveries of radioactivity from DLT- and InTn-labeled RSA were 5 and 6.5 days, respectively, again indicating that the larger glycoconjugate label residualized more efficiently in cells following protein degradation. (Lactitol) 2 -N-CH 2 -CH 2 -NH-fluroescein (DLF) was also coupled to ASF by reductive amination and recovered quantitatively in liver at 1 h post-injection. Native ASF was an effective competitor for clearance of DLF-ASF from the circulation. Fluorescent degradation products were retained in liver with a half-life of 1.2 days. Residualizing fluorescent labels should be useful for identification and sorting of cells active in the degradation of plasma proteins

  7. Impacts of Residual Surfactant on Tetrachloroethene (PCE) Degradation Following Pilot-Scale SEAR Treatment at a Chloroethene-Impacted Site

    Science.gov (United States)

    Ramsburg, C. A.; Abriola, L. M.; Pennell, K. D.; Löffler, F. E.; Gamache, M.; Petrovskis, E. A.

    2003-04-01

    A pilot-scale surfactant-enhanced aquifer remediation (SEAR) demonstration was completed during the summer of 2000 at the Bachman Road site (Oscoda, MI USA). For this test, an aqueous solution of 60 g/L Tween 80 (polyoxyethylene (20) sorbitan monooleate) was used to recover tetrachloroethene (PCE) from a suspected source zone, located underneath a former dry-cleaning facility. Tween 80 was selected for use based upon its demonstrated capacity to solubilize PCE, “food-grade” status, and biodegradative potential. Hydraulic control was maintained throughout the test, with 95% of the injected surfactant mass recovered by a single extraction well. Source-zone monitoring conducted 15 months after SEAR treatment revealed the presence of previously undetected volatile fatty acids (acetate and formate) and PCE degradation products (trichloroethene, cis-1,2-dichloroethene, trans-1,2-dichlorethene, and vinyl chloride), in conjunction with PCE concentration reductions of approximately two orders-of-magnitude. The detection of volatile fatty acids is relevant, as they are likely fermentation products of residual Tween 80. Microbial reductive dechlorination is limited by available electron donors, and microcosm studies demonstrated that both acetate and formate support reductively dechlorinating populations present at the oligotrophic Bachman Road site aquifer. Surfactant transport simulations, using a regional flow model developed for the site, were employed to determine appropriate down-gradient monitoring locations. Drive point samples taken 15 months post-treatment in the vicinity of the simulated residual surfactant plume, contained elevated concentrations of acetate and PCE daughter products. Ongoing efforts include continued site-monitoring, and microcosm studies to corroborate a causal relationship between Tween 80 fermentation and PCE dechlorination.

  8. Inulin-125I-tyramine, an improved residualizing label for studies on sites of catabolism of circulating proteins

    International Nuclear Information System (INIS)

    Maxwell, J.L.; Baynes, J.W.; Thorpe, S.R.

    1988-01-01

    Residualizing labels for protein, such as dilactitol-125I-tyramine (125I-DLT) and cellobiitol-125I-tyramine, have been used to identify the tissue and cellular sites of catabolism of long-lived plasma proteins, such as albumin, immunoglobulins, and lipoproteins. The radioactive degradation products formed from labeled proteins are relatively large, hydrophilic, resistant to lysosomal hydrolases, and accumulate in lysosomes in the cells involved in degradation of the carrier protein. However, the gradual loss of the catabolites from cells (t1/2 approximately 2 days) has limited the usefulness of residualizing labels in studies on longer lived proteins. We describe here a higher molecular weight (Mr approximately 5000), more efficient residualizing glycoconjugate label, inulin-125I-tyramine (125I-InTn). Attachment of 125I-InTn had no effect on the plasma half-life or tissue sites of catabolism of asialofetuin, fetuin, or rat serum albumin in the rat. The half-life for hepatic retention of degradation products from 125I-InTn-labeled asialofetuin was 5 days, compared to 2.3 days for 125I-DLT-labeled asialofetuin. The whole body half-lives for radioactivity from 125I-InTn-, 125I-DLT-, and 125I-labeled rat serum albumin were 7.5, 4.3, and 2.2 days, respectively. The tissue distribution of degradation products from 125I-InTn-labeled proteins agreed with results of previous studies using 125I-DLT, except that a greater fraction of total degradation products was recovered in tissues. Kinetic analyses indicated that the average half-life for retention of 125I-InTn degradation products in tissues is approximately 5 days and suggested that in vivo there are both slow and rapid routes for release of degradation products from cells

  9. Designed optimization of a single-step extraction of fucose-containing sulfated polysaccharides from Sargassum sp

    DEFF Research Database (Denmark)

    Ale, Marcel Tutor; Mikkelsen, Jørn Dalgaard; Meyer, Anne S.

    2012-01-01

    Fucose-containing sulfated polysaccharides can be extracted from the brown seaweed, Sargassum sp. It has been reported that fucose-rich sulfated polysaccharides from brown seaweeds exert different beneficial biological activities including anti-inflammatory, anticoagulant, and anti-viral effects....... Classical extraction of fucose-containing sulfated polysaccharides from brown seaweed species typically involves extended, multiple-step, hot acid, or CaCl2 treatments, each step lasting several hours. In this work, we systematically examined the influence of acid concentration (HCl), time, and temperature...... on the yield of fucosecontaining sulfated polysaccharides (FCSPs) in statistically designed two-step and single-step multifactorial extraction experiments. All extraction factors had significant effects on the fucose-containing sulfated polysaccharides yield, with the temperature and time exerting positive...

  10. Pathways analysis and radiation-dose estimates for radioactive residues at formerly utilized MED/AEC sites

    International Nuclear Information System (INIS)

    Gilbert, T.L.; Chee, P.C.; Knight, M.J.; Peterson, J.M.; Roberts, C.J.; Robinson, J.E.; Tsai, S.Y.H.; Yuan, Y.C.

    1983-03-01

    Methods of analysis are developed for estimating the largest individual radiation dose that could result from residual radioactivity at sites identified by the Formerly Utilized Sites Remedial Action Program (FUSRAP) of the US Department of Energy. Two unique aspects of the methods are (1) a systematic structuring of the radiation pathways analysis into source terms, source-to-exposure analysis, and exposure-to-dose analysis, and (2) the systematic use of data on the average concentrations of naturally occurring radionuclides in soil, food, and the human body in order to assess the validity of model calculations and obtain more realistic values. The methods are applied to a typical FUSRAP site in order to obtain generic source-to-dose (D/S) conversion factors for estimating the radiation dose to the maximally exposed individual from a known concentration of radionuclides in the soil. The D/S factors are used to derive soil guidelines, i.e., the limiting concentrations of radionuclides at a typical FUSRAP site that are unlikely to result in individual dose limits that exceed generally accepted radiation protection standards. The results lead to the conclusion that the soil guidelines should not exceed 17, 75, and 300 pCi/g for Ra-226, U-238, and Th-230, respectively

  11. Pathways analysis and radiation-dose estimates for radioactive residues at formerly utilized MED/AEC sites

    Energy Technology Data Exchange (ETDEWEB)

    Gilbert, T.L.; Chee, P.C.; Knight, M.J.; Peterson, J.M.; Roberts, C.J.; Robinson, J.E.; Tsai, S.Y.H.; Yuan, Y.C.

    1983-03-01

    Methods of analysis are developed for estimating the largest individual radiation dose that could result from residual radioactivity at sites identified by the Formerly Utilized Sites Remedial Action Program (FUSRAP) of the US Department of Energy. Two unique aspects of the methods are (1) a systematic structuring of the radiation pathways analysis into source terms, source-to-exposure analysis, and exposure-to-dose analysis, and (2) the systematic use of data on the average concentrations of naturally occurring radionuclides in soil, food, and the human body in order to assess the validity of model calculations and obtain more realistic values. The methods are applied to a typical FUSRAP site in order to obtain generic source-to-dose (D/S) conversion factors for estimating the radiation dose to the maximally exposed individual from a known concentration of radionuclides in the soil. The D/S factors are used to derive soil guidelines, i.e., the limiting concentrations of radionuclides at a typical FUSRAP site that are unlikely to result in individual dose limits that exceed generally accepted radiation protection standards. The results lead to the conclusion that the soil guidelines should not exceed 17, 75, and 300 pCi/g for Ra-226, U-238, and Th-230, respectively.

  12. Structural characterization of the PTS IIA and IIB proteins associated with pneumococcal fucose utilization.

    Science.gov (United States)

    Higgins, Melanie A; Hamilton, Aileen M; Boraston, Alisdair B

    2017-05-01

    Streptococcus pneumoniae harbors a significant number of transporters, including phosphotransferase (PTS) systems, allowing the bacterium to utilize a number of different carbohydrates for metabolic and other purposes. The genes encoding for one PTS transport system in particular (EII fuc ) are found within a fucose utilization operon in S. pneumoniae TIGR4. Here, we report the three-dimensional structures of IIA fuc and IIB fuc providing evidence that this PTS system belongs to the EII man family. Additionally, the predicted metabolic pathway for this distinctive fucose utilization system suggests that EII fuc transports the H-disaccharide blood group antigen, which would represent a novel PTS transporter specificity. Proteins 2017; 85:963-968. © 2016 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  13. Fucose-containing sulfated polysaccharides from brown seaweed: Extraction technolgy and bioactivity assessment

    DEFF Research Database (Denmark)

    Ale, Marcel Tutor

    will generate new valuable products that may help lessen coastal pollution by seaweeds and create new seaweed-based resources. Thus, utilization of these natural resources is of great importance. The objectives of this PhD study were to develop a technology to extract bioactive compounds from nuisance brown...... seaweeds, and investigate their bioactivity. To this effect, designed optimized extraction of fucose-containing sulfated polysaccharides (FCSPs) and/or crude fucoidan from brown seaweed were performed, and the bioactivity of the isolated FCSPs was investigated. Moreover, to assess the potential of seaweed...... to assimilate nitrogen-based nutrients, a technology for accurate monitoring of differential seaweed growth responses to nutrient assimilation was also developed. Fucoidan is a term used to describe a class of sulfated polysaccharides extracted from brown seaweed, which contains substantial amounts of fucose...

  14. Full scale amendment of a contaminated wood impregnation site with iron water treatment residues

    DEFF Research Database (Denmark)

    Nielsen, Sanne Skov; Kjeldsen, Peter; Jakobsen, Rasmus

    2016-01-01

    amendment a 100 m2 test site and a control site (without amendment) were monitored for 14 months. Also soil analysis of Fe to evaluate the degree of soil and Fe-WTR mixing was done. Stabilization with Fe-WTR had a significant effect on leachable contaminants, reducing pore water As by 93%, Cu by 91% and Cr...... by 95% in the upper samplers. Dosage and mixing of Fe-WTR in the soil proved to be difficult in the deeper part of the field, and pore water concentrations of arsenic was generally higher. Despite water logged conditions no increase in dissolved iron or arsenic was observed in the amended soil. Our...... field scale amendment of contaminated soil was overall successful in decreasing leaching of As, Cr and Cu.With minor improvements in the mixing and delivery strategy, this stabilization method is suggested for use in cases, where leaching of Cu, Cr and As constitutes a risk for groundwater...

  15. One-pot synthesis of GDP-l-fucose by a four-enzyme cascade expressed in Lactococcus lactis.

    Science.gov (United States)

    Li, Ling; Kim, Seul-Ah; Heo, Ji Eun; Kim, Tae-Jip; Seo, Jin-Ho; Han, Nam Soo

    2017-12-20

    GDP-l-fucose is an l-fucose donor to synthesize fucosylated compounds such as human milk oligosaccharides or Lewis antigen. In this study, we used Lactococcus lactis subsp. cremoris NZ9000 to express 4 enzymes, ManB, ManC, Gmd, and WcaG and produced GDP-l-fucose by using one-pot synthesis method with mannose-6-phosphate as substrate and the enzymes as biocatalyst. For preparation of enzyme mixture, 4 genes (manB, manC, gmd, and wcaG) cloned from Escherichia coli were transformed into L. lactis strains using pNZ8008 and the recombinant cell lysates were obtained after cultivation. When mannose-6-phosphate was used as the substrate, the consecutive reactions with ManB, ManC, Gmd, and WcaG resulted in the successful production of GDP-l-fucose (0.13mM). When GDP-d-mannose was used as the substrate, it was entirely converted to GDP-l-fucose (0.2mM; 0.12g/L) via 2 enzymatic reactions mediated by Gmd and WcaG. This is the first report of GDP-l-fucose production by using multiple enzymes expressed in lactic acid bacteria. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Potential of L-fucose isolated from Brown Seaweeds as Promising Natural Emulsifier compare to Carboxymethyl Cellulose (CMC)

    Science.gov (United States)

    Al-Baarri, A. N.; Legowo, A. M.; Widayat; Abduh, S. B. M.; Lestari, F. P.; Desnasari, D.; Santoso, I. P. M.

    2018-02-01

    L-fucose has been understood as sulfated polysaccharides and it could be extracted and fractionated from brown algae. These polysaccharides contains carbohydrate, sulfate, and protein that may be used as emulsifier. This research was aimed to study the emulsification properties of L-fucose through the determination of total dissolved solids (TDS), color CIE L*a*b* and stability of oil-in-water emulsion. As much as 0.5% of high concentrated L-fucose and 0.5% of carboxymethyl cellulose (CMC) were used as emulsifier in a 10% (v/v) oil-in-water (O/W) emulsion. The emulsifier was added to O/W emulsions and then heated at 72°C. Result of stability emulsion and TDS showed that L-fucose was comparable to the CMC but remarkable changed the color of O/W emulsion. Heating process significantly reduced the stability O/W emulsion when L-fucose was applied. As conclusion, L-fucose might be used as natural emulsifier in O/W emulsion but in the low heat treatment of food processing. This study may provide valuable information for utilizing natural emulsifier from abundant resources from nature.

  17. 2-Fluoro-L-Fucose Is a Metabolically Incorporated Inhibitor of Plant Cell Wall Polysaccharide Fucosylation.

    Directory of Open Access Journals (Sweden)

    Jose A Villalobos

    Full Text Available The monosaccharide L-fucose (L-Fuc is a common component of plant cell wall polysaccharides and other plant glycans, including the hemicellulose xyloglucan, pectic rhamnogalacturonan-I (RG-I and rhamnogalacturonan-II (RG-II, arabinogalactan proteins, and N-linked glycans. Mutations compromising the biosynthesis of many plant cell wall polysaccharides are lethal, and as a result, small molecule inhibitors of plant cell wall polysaccharide biosynthesis have been developed because these molecules can be applied at defined concentrations and developmental stages. In this study, we characterize novel small molecule inhibitors of plant fucosylation. 2-fluoro-L-fucose (2F-Fuc analogs caused severe growth phenotypes when applied to Arabidopsis seedlings, including reduced root growth and altered root morphology. These phenotypic defects were dependent upon the L-Fuc salvage pathway enzyme L-Fucose Kinase/ GDP-L-Fucose Pyrophosphorylase (FKGP, suggesting that 2F-Fuc is metabolically converted to the sugar nucleotide GDP-2F-Fuc, which serves as the active inhibitory molecule. The L-Fuc content of cell wall matrix polysaccharides was reduced in plants treated with 2F-Fuc, suggesting that this molecule inhibits the incorporation of L-Fuc into these polysaccharides. Additionally, phenotypic defects induced by 2F-Fuc treatment could be partially relieved by the exogenous application of boric acid, suggesting that 2F-Fuc inhibits RG-II biosynthesis. Overall, the results presented here suggest that 2F-Fuc is a metabolically incorporated inhibitor of plant cellular fucosylation events, and potentially suggest that other 2-fluorinated monosaccharides could serve as useful chemical probes for the inhibition of cell wall polysaccharide biosynthesis.

  18. 2-Fluoro-L-Fucose Is a Metabolically Incorporated Inhibitor of Plant Cell Wall Polysaccharide Fucosylation

    Science.gov (United States)

    Wallace, Ian S.

    2015-01-01

    The monosaccharide L-fucose (L-Fuc) is a common component of plant cell wall polysaccharides and other plant glycans, including the hemicellulose xyloglucan, pectic rhamnogalacturonan-I (RG-I) and rhamnogalacturonan-II (RG-II), arabinogalactan proteins, and N-linked glycans. Mutations compromising the biosynthesis of many plant cell wall polysaccharides are lethal, and as a result, small molecule inhibitors of plant cell wall polysaccharide biosynthesis have been developed because these molecules can be applied at defined concentrations and developmental stages. In this study, we characterize novel small molecule inhibitors of plant fucosylation. 2-fluoro-L-fucose (2F-Fuc) analogs caused severe growth phenotypes when applied to Arabidopsis seedlings, including reduced root growth and altered root morphology. These phenotypic defects were dependent upon the L-Fuc salvage pathway enzyme L-Fucose Kinase/ GDP-L-Fucose Pyrophosphorylase (FKGP), suggesting that 2F-Fuc is metabolically converted to the sugar nucleotide GDP-2F-Fuc, which serves as the active inhibitory molecule. The L-Fuc content of cell wall matrix polysaccharides was reduced in plants treated with 2F-Fuc, suggesting that this molecule inhibits the incorporation of L-Fuc into these polysaccharides. Additionally, phenotypic defects induced by 2F-Fuc treatment could be partially relieved by the exogenous application of boric acid, suggesting that 2F-Fuc inhibits RG-II biosynthesis. Overall, the results presented here suggest that 2F-Fuc is a metabolically incorporated inhibitor of plant cellular fucosylation events, and potentially suggest that other 2-fluorinated monosaccharides could serve as useful chemical probes for the inhibition of cell wall polysaccharide biosynthesis. PMID:26414071

  19. Biosynthesis of fucose containing lacto-series glycolipids in human colonic adenocarcinoma Colo 205 cells.

    Science.gov (United States)

    Holmes, E H; Levery, S B

    1989-11-01

    Biosynthesis of fucose containing lacto-series glycolipids has been studied in human colonic adenocarcinoma Colo 205 cells. Transfer of fucose in both alpha 1----3 linkage to type 2 chain acceptors and alpha 1----4 linkage to type 1 chain acceptors was demonstrated with a Triton X-100 solubilized membrane fraction. The enzyme was found to be highly active over a broad pH range between 6.0 and 7.5. Kinetics of the transfer reactions were studied and indicated that the enzyme had an apparent Km for GDPfucose of 53 and 49 microM with acceptors nLc4 and Lc4, respectively. The apparent Km values for acceptors Lc4, nLc4, and IV3NeuAcnLc4 were determined to be 42, 18, and 26 microM, respectively. Transfer of fucose to the type 1 chain acceptor Lc4 alone and in the presence of increasing concentrations of the type 2 chain acceptor IV3NeuAcnLc4 or Gb3 suggested that both type 1 and 2 acceptors were alternate acceptors for a single enzyme. This was further established by the finding that IV3NeuAcnLc4 behaved as a competitive inhibitor of fucose transfer with respect to Lc4. Conditions were defined for preparative scale in vitro synthesis of fucosylated products of nLc6 catalyzed by the Colo 205 cell enzyme. Yields of the monofucosyl derivative of 2.5 mg (46%) and 1 mg (17%) of the difucosyl derivative were obtained from 5 mg of original nLc6. The structures of these biosynthetic products were carefully studied by 1H NMR, +FAB-MS, and methylation analysis. These studies revealed extremely high purity products composed of III3FucnLc6 and III3V3Fuc2nLc6. The significance of the nature of these products and enzymatic properties is discussed.

  20. Identification of Key Functional Residues in the Active Site of Human β1,4-Galactosyltransferase 7

    Science.gov (United States)

    Talhaoui, Ibtissam; Bui, Catherine; Oriol, Rafael; Mulliert, Guillermo; Gulberti, Sandrine; Netter, Patrick; Coughtrie, Michael W. H.; Ouzzine, Mohamed; Fournel-Gigleux, Sylvie

    2010-01-01

    Glycosaminoglycans (GAGs) play a central role in many pathophysiological events, and exogenous xyloside substrates of β1,4-galactosyltransferase 7 (β4GalT7), a major enzyme of GAG biosynthesis, have interesting biomedical applications. To predict functional peptide regions important for substrate binding and activity of human β4GalT7, we conducted a phylogenetic analysis of the β1,4-galactosyltransferase family and generated a molecular model using the x-ray structure of Drosophila β4GalT7-UDP as template. Two evolutionary conserved motifs, 163DVD165 and 221FWGWGREDDE230, are central in the organization of the enzyme active site. This model was challenged by systematic engineering of point mutations, combined with in vitro and ex vivo functional assays. Investigation of the kinetic properties of purified recombinant wild-type β4GalT7 and selected mutants identified Trp224 as a key residue governing both donor and acceptor substrate binding. Our results also suggested the involvement of the canonical carboxylate residue Asp228 acting as general base in the reaction catalyzed by human β4GalT7. Importantly, ex vivo functional tests demonstrated that regulation of GAG synthesis is highly responsive to modification of these key active site amino acids. Interestingly, engineering mutants at position 224 allowed us to modify the affinity and to modulate the specificity of human β4GalT7 toward UDP-sugars and xyloside acceptors. Furthermore, the W224H mutant was able to sustain decorin GAG chain substitution but not GAG synthesis from exogenously added xyloside. Altogether, this study provides novel insight into human β4GalT7 active site functional domains, allowing manipulation of this enzyme critical for the regulation of GAG synthesis. A better understanding of the mechanism underlying GAG assembly paves the way toward GAG-based therapeutics. PMID:20843813

  1. Baseline risk assessment for the quarry residuals operable unit of the Weldon Spring Site, Weldon Spring, Missouri

    International Nuclear Information System (INIS)

    1998-02-01

    The U.S. Department of Energy (DOE) is conducting cleanup activities at the Weldon Spring site, located in St. Charles County, Missouri, about 48 km (30 mi) west of St. Louis. Cleanup of the site consists of several integrated components. The quarry residuals operable unit (QROU), consisting of the Weldon Spring quarry and its surrounding area, is one of four operable units being evaluated. In accordance with requirements of the Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA), as amended, DOE is conducting a remedial investigation/feasibility study (RI/FS) to determine the proper response to address various contaminated media that constitute the QROU. Specifically, the operable unit consists of the following areas and media: the residual material remaining at the Weldon Spring quarry after removal of the pond water and the bulk waste; groundwater underlying the quarry and surrounding area; and other media located in the surrounding vicinity of the quarry, including surface water and sediment at Femme Osage Slough, Little Femme Osage Creek, and Femme Osage Creek. An initial evaluation of conditions at the quarry area identified remaining data requirements needed to support the conceptual site exposure and hydrogeological models. These data requirements are discussed in the RI/FS work plan issued in January 1994. Soil contamination located at a property adjacent to the quarry, referred to as Vicinity Property 9 (VP9), was originally part of the scope of the QROU, as discussed in the work plan. However, a decision was subsequently made to remediate this vicinity property as part of cleanup activities for the chemical plant operable unit, as provided for in the Record of Decision (ROD). Remediation of VP9 was completed in early 1996. Hence, this baseline risk assessment (BRA) does not address VP9

  2. Baseline risk assessment for the quarry residuals operable unit of the Weldon Spring Site, Weldon Spring, Missouri

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1998-02-01

    The U.S. Department of Energy (DOE) is conducting cleanup activities at the Weldon Spring site, located in St. Charles County, Missouri, about 48 km (30 mi) west of St. Louis. Cleanup of the site consists of several integrated components. The quarry residuals operable unit (QROU), consisting of the Weldon Spring quarry and its surrounding area, is one of four operable units being evaluated. In accordance with requirements of the Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA), as amended, DOE is conducting a remedial investigation/feasibility study (RI/FS) to determine the proper response to address various contaminated media that constitute the QROU. Specifically, the operable unit consists of the following areas and media: the residual material remaining at the Weldon Spring quarry after removal of the pond water and the bulk waste; groundwater underlying the quarry and surrounding area; and other media located in the surrounding vicinity of the quarry, including surface water and sediment at Femme Osage Slough, Little Femme Osage Creek, and Femme Osage Creek. An initial evaluation of conditions at the quarry area identified remaining data requirements needed to support the conceptual site exposure and hydrogeological models. These data requirements are discussed in the RI/FS work plan issued in January 1994. Soil contamination located at a property adjacent to the quarry, referred to as Vicinity Property 9 (VP9), was originally part of the scope of the QROU, as discussed in the work plan. However, a decision was subsequently made to remediate this vicinity property as part of cleanup activities for the chemical plant operable unit, as provided for in the Record of Decision (ROD). Remediation of VP9 was completed in early 1996. Hence, this baseline risk assessment (BRA) does not address VP9.

  3. Submicroscopic pores grafted using the residual sites produced by swift heavy ions

    International Nuclear Information System (INIS)

    Mazzei, R.; Betz, N.; Bermudez, G. Garcia; Massa, G.; Smolko, E.

    2005-01-01

    To produce nuclear track membranes (NTM) with submicroscopic pores poly(vinylidene difluoride) (PVDF) foils were irradiated with Cl, Ag and Pb ions. Then they were chemically etched for different times and grafted with acrylic acid. The grafting yields were determined by weight measurements as a function of ion fluence, etching time and also analysed using Fourier transform infrared spectroscopy. Both measurements suggest that the acrylic acid was grafted on the pore wall of the NTM using the active sites left by the ion beam

  4. European sites contaminated by residues from the ore extracting and processing industries

    International Nuclear Information System (INIS)

    Vandenhove, H.

    2000-01-01

    Activities linked with the ore extraction and processing industries may lead to enhanced levels of naturally occurring radionuclides (NORs) in products, by-products and waste and at the installations and in the surroundings of the facility. In the framework of the EC-DGXI CARE project (Common Approach for REstoration of contaminated sites) nine important categories of industries were identified and discussions were summarized on the industrial processes and the levels of NORs in parent material, waste and by-products. The most contaminating industries are uranium mining and milling, metal mining and smelting and the phosphate industry. Radionuclide levels in products and/or waste products from the oil and gas extraction industry and of the rare earth, zirconium and ceramics industries may be particularly elevated, but waste streams are limited. The impact on the public from coal mining and power production from coal is commonly considered low. No typical values are available for contaminant levels in materials, buildings and surroundings of radium extraction and luminizing plants, nor for thorium extraction and processing plants. An attempt to give an overview of sites in Europe contaminated with NORs, with emphasis on past practices, was only partly successful since information was often limited or unavailable. The most prominent case of environmental contamination due to mining and processing activities (uranium, metal and coal mining) is in eastern Germany. (author)

  5. Evidence for the existence of a tyrosyl residue in the nicotinamide adenine dinucleotide binding site of chicken liver xanthine dehydrogenase

    International Nuclear Information System (INIS)

    Nishino, T.; Nishino, T.

    1987-01-01

    Xanthine-NAD and NADH-methylene blue oxidoreductase activities of chicken liver xanthine dehydrogenase were inactivated by incubation with 5'-[p-(fluorosulfonyl)benzoyl]adenosine (5'-FSBA), an active site directed reagent for nucleotide binding sites. The inactivation reaction displayed pseudo-first-order kinetics. A double-reciprocal plot of inactivation velocity vs. 5'-FSBA concentration showed that 5'-FSBA and enzyme formed a complex prior to inactivation. NAD protected the enzyme from inactivation by 5'-FSBA in a competitive fashion. The modified enzyme had the same xanthine-dichlorophenolindophenol and xanthine-O 2 oxidoreductase activities as the native enzyme, and on addition of xanthine to the modified enzyme, bleaching of the spectrum occurred in the visible region. The amount of radioactivity incorporated into the enzyme by incubation with [ 14 C]-5'-FSBA was parallel to the loss of xanthine-NAD oxidoreductase activity, and the stoichiometry was 1 mol/mol of enzyme-bound FAD for complete inactivation. These results indicated that 5'-FSBA modified specifically the binding site for NAD of chicken liver xanthine dehydrogenase. The incorporated radioactivity was released slowly from 14 C-labeled enzyme by incubation with dithiothreitol with concomitant restoration of catalytic activity. The modified residue responsible for inactivation was identified as a tyrosine

  6. Derivation of residual radioactive material guidelines for uranium in soil at the Former Associate Aircraft Tool and Manufacturing Company Site, Fairfield, Ohio

    International Nuclear Information System (INIS)

    Faillace, E.R.; Nimmagadda, M.; Yu, C.

    1995-01-01

    Residual radioactive material guidelines for uranium in soil were derived for the former Associate Aircraft Tool and Manufacturing Company site in Fairfield, Ohio. This site has been identified for remedial action under the U.S. Department of Energy's (DOE's) Formerly Utilized Sites Remedial Action Program (FUSRAP). Single-nuclide and total-uranium guidelines were derived on the basis of the requirement that, after remedial action, the 50-year committed effective dose equivalent to a hypothetical individual living or working in the immediate vicinity of the site should not exceed (1) 30 mrem/yr for the current-use and likely future-use scenarios or (2) 100 mrem/yr for less likely future-use scenarios. The DOE residual radioactive material (RESRAD) computer code, which implements the methodology described in the DOE manual for establishing residual radioactive material guidelines, was used in this evaluation

  7. A single acidic residue can guide binding site selection but does not govern QacR cationic-drug affinity.

    Directory of Open Access Journals (Sweden)

    Kate M Peters

    Full Text Available Structures of the multidrug-binding repressor protein QacR with monovalent and bivalent cationic drugs revealed that the carboxylate side-chains of E90 and E120 were proximal to the positively charged nitrogens of the ligands ethidium, malachite green and rhodamine 6G, and therefore may contribute to drug neutralization and binding affinity. Here, we report structural, biochemical and in vivo effects of substituting these glutamate residues. Unexpectedly, substitutions had little impact on ligand affinity or in vivo induction capabilities. Structures of QacR(E90Q and QacR(E120Q with ethidium or malachite green took similar global conformations that differed significantly from all previously described QacR-drug complexes but still prohibited binding to cognate DNA. Strikingly, the QacR(E90Q-rhodamine 6G complex revealed two mutually exclusive rhodamine 6G binding sites. Despite multiple structural changes, all drug binding was essentially isoenergetic. Thus, these data strongly suggest that rather than contributing significantly to ligand binding affinity, the role of acidic residues lining the QacR multidrug-binding pocket is primarily to attract and guide cationic drugs to the "best available" positions within the pocket that elicit QacR induction.

  8. Transforming growth factor β signaling upregulates the expression of human GDP-fucose transporter by activating transcription factor Sp1.

    Science.gov (United States)

    Xu, Yu-Xin; Ma, Anna; Liu, Li

    2013-01-01

    GDP-fucose transporter plays a crucial role in fucosylation of glycoproteins by providing activated fucose donor, GDP-fucose, for fucosyltransferases in the lumen of the Golgi apparatus. Fucose-containing glycans are involved in many biological processes, which are essential for growth and development. Mutations in the GDP-fucose transporter gene cause leukocyte adhesion deficiency syndrome II, a disease characterized by slow growth, mental retardation and immunodeficiency. However, no information is available regarding its transcriptional regulation. Here, by using human cells, we show that TGF-β1 specifically induces the GDP-fucose transporter expression, but not other transporters tested such as CMP-sialic acid transporter, suggesting a diversity of regulatory pathways for the expression of these transporters. The regulatory elements that are responsive to the TGF-β1 stimulation are present in the region between bp -330 and -268 in the GDP-fucose transporter promoter. We found that this region contains two identical octamer GC-rich motifs (GGGGCGTG) that were demonstrated to be essential for the transporter expression. We also show that the transcription factor Sp1 specifically binds to the GC-rich motifs in vitro and Sp1 coupled with phospho-Smad2 is associated with the promoter region covering the Sp1-binding motifs in vivo using chromatin immunoprecipitation (ChIP) assays. In addition, we further confirmed that Sp1 is essential for the GDP-fucose transporter expression stimulated by TGF-β1 using a luciferase reporter system. These results highlight the role of TGF-β signaling in regulation of the GDP-fucose transporter expression via activating Sp1. This is the first transcriptional study for any nucleotide sugar transporters that have been identified so far. Notably, TGF-β1 receptor itself is known to be modified by fucosylation. Given the essential role of GDP-fucose transporter in fucosylation, the finding that TGF-β1 stimulates the expression of

  9. DAF in diabetic patients is subject to glycation/inactivation at its active site residues.

    Science.gov (United States)

    Flückiger, Rudolf; Cocuzzi, Enzo; Nagaraj, Ram H; Shoham, Menachem; Kern, Timothy S; Medof, M Edward

    2018-01-01

    Decay accelerating factor (DAF or CD55) is a cell associated C3 and C5 convertase regulator originally described in terms of protection of self-cells from systemic complement but now known to modulate adaptive T cell responses. It is expressed on all cell types. We investigated whether nonenzymatic glycation could impair its function and potentially be relevant to complications of diabetes mellitus and other conditions that result in nonenzymatic glycation including cancer, Alzheimer's disease, and aging. Immunoblots of affinity-purified DAF from erythrocytes of patients with diabetes showed pentosidine, glyoxal-AGEs, carboxymethyllysine, and argpyrimidine. HPLC/MS analyses of glucose modified DAF localized the sites of AGE modifications to K 125 adjacent to K 126 , K 127 at the junction of CCPs2-3 and spatially near R 96 , and R 100 , all identified as being critical for DAF's function. Functional analyses of glucose or ribose treated DAF protein showed profound loss of its regulatory activity. The data argue that de-regulated activation of systemic complement and de-regulated activation of T cells and leukocytes could result from non-enzymatic glycation of DAF. Copyright © 2017. Published by Elsevier Ltd.

  10. Site-directed mutagenesis under the direction of in silico protein docking modeling reveals the active site residues of 3-ketosteroid-Δ1-dehydrogenase from Mycobacterium neoaurum.

    Science.gov (United States)

    Qin, Ning; Shen, Yanbing; Yang, Xu; Su, Liqiu; Tang, Rui; Li, Wei; Wang, Min

    2017-07-01

    3-Ketosteroid-Δ 1 -dehydrogenases (KsdD) from Mycobacterium neoaurum could transform androst-4-ene-3,17-dione (AD) to androst-1,4-diene-3,17-dione. This reaction has a significant effect on the product of pharmaceutical steroid. The crystal structure and active site residues information of KsdD from Mycobacterium is not yet available, which result in the engineering of KsdD is tedious. In this study, by the way of protein modeling and site-directed mutagenesis, we find that, Y122, Y125, S138, E140 and Y541 from the FAD-binding domain and Y365 from the catalytic domain play a key role in this transformation. Compared with the wild type, the decline in AD conversion for mutants illustrated that Y125, Y365, and Y541 were essential to the function of KsdD. Y122, S138 and E140 contributed to the catalysis of KsdD. The following analysis revealed the catalysis mechanism of these mutations in KsdD of Mycobacterium. These information presented here facilitate the manipulation of the catalytic properties of the enzyme to improve its application in the pharmaceutical steroid industry.

  11. Lectin histochemistry of goblet cell sugar residues in the gut of the chick embryo and of the newborn.

    Science.gov (United States)

    Bryk, S G; Sgambati, E; Gheri Bryk, G

    1999-04-01

    The anlage of duodenum, ileum and colon were removed from chick embryos of day 8-21 of incubation and from 1-day-old chicks. A battery of seven different horseradish peroxidase-conjugated lectins (PNA, SBA, DBA, Con A, WGA, LTA and UEAI) was used to study the carbohydrate residues of the glycoconjugates in the goblet cells of the three parts of the intestine. The main results can be summarized as follows: differences in lectin binding were absent in the proximal and distal parts of the duodenum, ileum and colon. Lectin histochemistry showed differences among the three intestinal segments for the time of appearance of the oligosaccharides in the goblet mucus. In the colonic goblet cells of 1-day-old chicks, LTA and UEAI lectins showed two different types of linkage of alpha-L-fucose. This is the first demonstration of UEAI reactive sites in Gallus domesticus.

  12. Contribution of tryptophan residues to the combining site of a monoclonal anti dinitrophenyl spin-label antibody

    International Nuclear Information System (INIS)

    Anglister, J.; Bond, M.W.; Frey, T.; Leahy, D.; Levitt, M.; McConnell, H.M.; Rule, G.S.; Tomasello, J.; Whittaker, M.

    1987-01-01

    Two Fab fragments of the monoclonal anti dinitrophenyl (DNP) spin-label antibody AN02 were prepared by recombination of specifically deuterated heavy and light chains. In the recombinant H(I)L(II) all the tyrosines and phenylalanines were perdeuterated as were the tryptophan residues of the heavy chain. In the recombinant H(II)L(I) all the tyrosines and phenylalanines were perdeuterated as were the tryptophan residues of the light chain. Saturation of three resonances of H(I)L(II), assigned to tryptophan protons of the light chain, resulted in magnetization transfer to the aromatic proton at position 6 of the DNP ring and to the CH2 protons of the glycines linked to the DNP in a diamagnetic hapten (DNP-DG). Saturation of three resonances of H(II)L(I) assigned to tryptophan protons of the heavy chain resulted in magnetization transfer to the CH2 protons of the glycines in DNP-DG. From the dependence of the magnetization transfer on the irradiation time, the cross relaxation rates between the involved protons were estimated. The inferred distances between these protons of the hapten and certain tryptophan protons are 3-4 A. It is concluded that in the combining site of AN02 there is one tryptophan from the light chain and one tryptophan from the heavy chain that are very near the hapten. When all tyrosines and phenylalanines were perdeuterated and all tryptophan aromatic protons were deuterated except for the protons at positions 2 and 5, titration of the Fab fragments with variable amounts of paramagnetic hapten showed that one proton from the light chain tryptophan is near (less than 7 A) the unpaired electron and that three other protons are significantly closer than 15 A

  13. Contribution of tryptophan residues to the combining site of a monoclonal anti dinitrophenyl spin-label antibody

    Energy Technology Data Exchange (ETDEWEB)

    Anglister, J.; Bond, M.W.; Frey, T.; Leahy, D.; Levitt, M.; McConnell, H.M.; Rule, G.S.; Tomasello, J.; Whittaker, M.

    1987-09-22

    Two Fab fragments of the monoclonal anti dinitrophenyl (DNP) spin-label antibody AN02 were prepared by recombination of specifically deuterated heavy and light chains. In the recombinant H(I)L(II) all the tyrosines and phenylalanines were perdeuterated as were the tryptophan residues of the heavy chain. In the recombinant H(II)L(I) all the tyrosines and phenylalanines were perdeuterated as were the tryptophan residues of the light chain. Saturation of three resonances of H(I)L(II), assigned to tryptophan protons of the light chain, resulted in magnetization transfer to the aromatic proton at position 6 of the DNP ring and to the CH2 protons of the glycines linked to the DNP in a diamagnetic hapten (DNP-DG). Saturation of three resonances of H(II)L(I) assigned to tryptophan protons of the heavy chain resulted in magnetization transfer to the CH2 protons of the glycines in DNP-DG. From the dependence of the magnetization transfer on the irradiation time, the cross relaxation rates between the involved protons were estimated. The inferred distances between these protons of the hapten and certain tryptophan protons are 3-4 A. It is concluded that in the combining site of AN02 there is one tryptophan from the light chain and one tryptophan from the heavy chain that are very near the hapten. When all tyrosines and phenylalanines were perdeuterated and all tryptophan aromatic protons were deuterated except for the protons at positions 2 and 5, titration of the Fab fragments with variable amounts of paramagnetic hapten showed that one proton from the light chain tryptophan is near (less than 7 A) the unpaired electron and that three other protons are significantly closer than 15 A.

  14. Metabolic engineering of Escherichia coli to produce 2'-fucosyllactose via salvage pathway of guanosine 5'-diphosphate (GDP)-l-fucose.

    Science.gov (United States)

    Chin, Young-Wook; Seo, Nari; Kim, Jae-Han; Seo, Jin-Ho

    2016-11-01

    2'-Fucosyllactose (2-FL) is one of the key oligosaccharides in human milk. In the present study, the salvage guanosine 5'-diphosphate (GDP)-l-fucose biosynthetic pathway from fucose was employed in engineered Escherichia coli BL21star(DE3) for efficient production of 2-FL. Introduction of the fkp gene coding for fucokinase/GDP-l-fucose pyrophosphorylase (Fkp) from Bacteroides fragilis and the fucT2 gene encoding α-1,2-fucosyltransferase from Helicobacter pylori allows the engineered E. coli to produce 2-FL from fucose, lactose and glycerol. To enhance the lactose flux to 2-FL production, the attenuated, and deleted mutants of β-galactosidase were employed. Moreover, the 2-FL yield and productivity were further improved by deletion of the fucI-fucK gene cluster coding for fucose isomerase (FucI) and fuculose kinase (FucK). Finally, fed-batch fermentation of engineered E. coli BL21star(DE3) deleting lacZ and fucI-fucK, and expressing fkp and fucT2 resulted in 23.1 g/L of extracellular concentration of 2-FL and 0.39 g/L/h productivity. Biotechnol. Bioeng. 2016;113: 2443-2452. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  15. Assessment of alternatives for long-term management of uranium ore residues and contaminated soils located at DOE's Niagara Falls Storage Site

    International Nuclear Information System (INIS)

    Merry-Libby, P.

    1985-01-01

    About 11,000 m 3 of uranium ore residues and 180,000 m 3 of slightly contaminated soils (wastes) are consolidated within a diked containment area at the Niagara Falls Storage Site located about 30 km north of Buffalo, New York. The residues account for less than 6% of the total volume of contaminated materials but almost 99% of the radioactivity. The average radium-226 concentration in the residues is 67,000 pCi/g. The US Department of Energy is considering several alternatives for long-term management of the wastes and residues, including: improvement of the containment at NFSS, modification of the form of the residues, management of the residues separately from the wastes, management of the wastes and residues at another humid site (Oak Ridge, Tennessee) or an arid site (Hanford, Washington), and dispersal of the wastes in the ocean. Potential radiological risks associated with implementation of any of the alternatives are expected to be smaller than the nonradiological risks of occupational and transportation-related injuries and deaths. Dispersal of the slightly contaminated wastes in the ocean is not expected to result in any significant radiological risk to humans. The residues and wastes will remain hazardous for thousands of years. After controls cease, the radioactive materials will eventually be dispersed in the environment. Loss of the earthen covers over the buried materials is predicted to occur from several hundred to more than two million years, depending primarily on the use of the land surface. Groundwater will eventually be contaminated in all alternatives; however, the groundwater pathway is relatively insignificant with respect to radiological risks to the general population. 2 references, 2 figures, 6 tables

  16. Male-like sexual behavior of female mouse lacking fucose mutarotase

    Directory of Open Access Journals (Sweden)

    Lim Dae-sik

    2010-07-01

    Full Text Available Abstract Background Mutarotases are recently characterized family of enzymes that are involved in the anomeric conversions of monosaccharides. The mammalian fucose mutarotase (FucM was reported in cultured cells to facilitate fucose utilization and incorporation into protein by glycosylation. However, the role of this enzyme in animal has not been elucidated. Results We generated a mutant mouse specifically lacking the fucose mutarotase (FucM gene. The FucM knockout mice displayed an abnormal sexual receptivity with a drastic reduction in lordosis score, although the animals were fertile due to a rare and forced intromission by a typical male. We examined the anteroventral periventricular nucleus (AVPv of the preoptic region in brain and found that the mutant females showed a reduction in tyrosine hydoxylase positive neurons compared to that of a normal female. Furthermore, the mutant females exhibited a masculine behavior, such as mounting to a normal female partner as well as showing a preference to female urine. We found a reduction of fucosylated serum alpha-fetoprotein (AFP in a mutant embryo relative to that of a wild-type embryo. Conclusions The observation that FucM-/- female mouse exhibits a phenotypic similarity to a wild-type male in terms of its sexual behavior appears to be due to the neurodevelopmental changes in preoptic area of mutant brain resembling a wild-type male. Since the previous studies indicate that AFP plays a role in titrating estradiol that are required to consolidate sexual preference of female mice, we speculate that the reduced level of AFP in FucM-/- mouse, presumably resulting from the reduced fucosylation, is responsible for the male-like sexual behavior observed in the FucM knock-out mouse.

  17. L-fucose in crystal structures of IgG-Fc: Reinterpretation of experimental data

    Czech Academy of Sciences Publication Activity Database

    Kolenko, Petr; Skálová, Tereza; Dohnálek, Jan; Hašek, Jindřich

    2008-01-01

    Roč. 73, č. 5 (2008), s. 608-615 ISSN 0010-0765 R&D Projects: GA ČR GA305/07/1073 Grant - others:FP6-2005-LifeSciHealth-6(EU) LSHG-CT-2006-031220 Institutional research plan: CEZ:AV0Z40500505 Source of funding: R - rámcový projekt EK Keywords : immunoglobulin * Fc-fragment * fucose Subject RIV: CD - Macromolecular Chemistry Impact factor: 0.784, year: 2008

  18. Allowable residual contamination levels for decommissioning the 115-F and 117-F facilities at the Hanford Site

    International Nuclear Information System (INIS)

    Kennedy, W.E. Jr.; Napier, B.A.

    1983-07-01

    This report contains the results of a study sponsored by UNC Nuclear Industries to determine Allowable Residual Contamination Levels (ARCL) for the 115-F and 117-F facilities at the Hanford Site. The purpose of this study is to provide data useful to UNC engineers in conducting safety and cost comparisons for decommissioning alternatives. The ARCL results are based on a scenario/exposure-pathway analysis and compliance with an annual dose limit for three specific modes of future use of the land and facilities. These modes of use are restricted, controlled, and unrestricted. Information on restricted and controlled use is provided to permit a full consideration of decommissioning alternatives. Procedures are presented for modifying the ARCL values to accommodate changes in the radionuclide mixture or concentrations and to determine instrument responses for various mixtures of radionuclides. Finally, a comparison is made between existing decommissioning guidance and the ARCL values calculated for unrestricted release of the 115-F and 117-F facilities. The comparison shows a good agreement

  19. Variation in one residue associated with the metal ion-dependent adhesion site regulates αIIbβ3 integrin ligand binding affinity.

    Directory of Open Access Journals (Sweden)

    Joel Raborn

    Full Text Available The Asp of the RGD motif of the ligand coordinates with the β I domain metal ion dependent adhesion site (MIDAS divalent cation, emphasizing the importance of the MIDAS in ligand binding. There appears to be two distinct groups of integrins that differ in their ligand binding affinity and adhesion ability. These differences may be due to a specific residue associated with the MIDAS, particularly the β3 residue Ala(252 and corresponding Ala in the β1 integrin compared to the analogous Asp residue in the β2 and β7 integrins. Interestingly, mutations in the adjacent to MIDAS (ADMIDAS of integrins α4β7 and αLβ2 increased the binding and adhesion abilities compared to the wild-type, while the same mutations in the α2β1, α5β1, αVβ3, and αIIbβ3 integrins demonstrated decreased ligand binding and adhesion. We introduced a mutation in the αIIbβ3 to convert this MIDAS associated Ala(252 to Asp. By combination of this mutant with mutations of one or two ADMIDAS residues, we studied the effects of this residue on ligand binding and adhesion. Then, we performed molecular dynamics simulations on the wild-type and mutant αIIbβ3 integrin β I domains, and investigated the dynamics of metal ion binding sites in different integrin-RGD complexes. We found that the tendency of calculated binding free energies was in excellent agreement with the experimental results, suggesting that the variation in this MIDAS associated residue accounts for the differences in ligand binding and adhesion among different integrins, and it accounts for the conflicting results of ADMIDAS mutations within different integrins. This study sheds more light on the role of the MIDAS associated residue pertaining to ligand binding and adhesion and suggests that this residue may play a pivotal role in integrin-mediated cell rolling and firm adhesion.

  20. D-arabinose metabolism in Escherichia coli B: induction and cotransductional mapping of the L-fucose-D-arabinose pathway enzymes.

    Science.gov (United States)

    Elsinghorst, E A; Mortlock, R P

    1988-12-01

    D-Arabinose is degraded by Escherichia coli B via some of the L-fucose pathway enzymes and a D-ribulokinase which is distinct from the L-fuculokinase of the L-fucose pathway. We found that L-fucose and D-arabinose acted as the apparent inducers of the enzymes needed for their degradation. These enzymes, including D-ribulokinase, appeared to be coordinately regulated, and mutants which constitutively synthesized the L-fucose enzymes also constitutively synthesized D-ribulokinase. In contrast to D-arabinose-positive mutants of E. coli K-12, in which L-fuculose-1-phosphate and D-ribulose-1-phosphate act as inducers of the L-fucose pathway, we found that these intermediates did not act as inducers in E. coli B. To further characterize the E. coli B system, some of the L-fucose-D-arabinose genes were mapped by using bacteriophage P1 transduction. A transposon Tn10 insertion near the E. coli B L-fucose regulon was used in two- and three-factor reciprocal crosses. The gene encoding D-ribulokinase, designated darK, was found to map within the L-fucose regulon, and the partial gene order was found to be Tn10-fucA-darK-fucI-fucK-thyA.

  1. Measurement of the residual radiation intensity at the Hiroshima and Nagasaki atomic bomb sites. Penetration of weapons radiation: application to the Hiroshima and Nagasaki studies

    Energy Technology Data Exchange (ETDEWEB)

    Pace, N; Smith, R E; Ritchie, R H; Hurst, G S

    1959-01-01

    This document contains 2 reports. The first is on the measurement of residual radiation intensity at the Hiroshima and Nagasaki bomb sites, the second is on the penetration of weapons radiation at Hiroshima and Nagasaki. Separate abstracts have been prepared for each report for inclusion in the Energy Database. (DMC)

  2. Radiological dose assessment for residual radioactive material in soil at the clean slate sites 1, 2, and 3, Tonopah Test Range

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1997-06-01

    A radiological dose assessment has been performed for Clean Slate Sites 1, 2, and 3 at the Tonopah Test Range, approximately 390 kilometers (240 miles) northwest of Las Vegas, Nevada. The assessment demonstrated that the calculated dose to hypothetical individuals who may reside or work on the Clean Slate sites, subsequent to remediation, does not exceed the limits established by the US Department of Energy for protection of members of the public and the environment. The sites became contaminated as a result of Project Roller Coaster experiments conducted in 1963 in support of the US Atomic Energy Commission (Shreve, 1964). Remediation of Clean Slate Sites 1, 2, and 3 is being performed to ensure that the 50-year committed effective dose equivalent to a hypothetical individual who lives or works on a Clean Slate site should not exceed 100 millirems per year. The DOE residual radioactive material guideline (RESRAD) computer code was used to assess the dose. RESRAD implements the methodology described in the DOE manual for establishing residual radioactive material guidelines (Yu et al., 1993a). In May and June of 1963, experiments were conducted at Clean Slate Sites 1, 2, and 3 to study the effectiveness of earth-covered structures for reducing the dispersion of nuclear weapons material as a result of nonnuclear explosions. The experiments required the detonation of various simulated weapons using conventional chemical explosives (Shreve, 1964). The residual radioactive contamination in the surface soil consists of weapons grade plutonium, depleted uranium, and their radioactive decay products.

  3. Radiological dose assessment for residual radioactive material in soil at the clean slate sites 1, 2, and 3, Tonopah Test Range

    International Nuclear Information System (INIS)

    1997-06-01

    A radiological dose assessment has been performed for Clean Slate Sites 1, 2, and 3 at the Tonopah Test Range, approximately 390 kilometers (240 miles) northwest of Las Vegas, Nevada. The assessment demonstrated that the calculated dose to hypothetical individuals who may reside or work on the Clean Slate sites, subsequent to remediation, does not exceed the limits established by the US Department of Energy for protection of members of the public and the environment. The sites became contaminated as a result of Project Roller Coaster experiments conducted in 1963 in support of the US Atomic Energy Commission (Shreve, 1964). Remediation of Clean Slate Sites 1, 2, and 3 is being performed to ensure that the 50-year committed effective dose equivalent to a hypothetical individual who lives or works on a Clean Slate site should not exceed 100 millirems per year. The DOE residual radioactive material guideline (RESRAD) computer code was used to assess the dose. RESRAD implements the methodology described in the DOE manual for establishing residual radioactive material guidelines (Yu et al., 1993a). In May and June of 1963, experiments were conducted at Clean Slate Sites 1, 2, and 3 to study the effectiveness of earth-covered structures for reducing the dispersion of nuclear weapons material as a result of nonnuclear explosions. The experiments required the detonation of various simulated weapons using conventional chemical explosives (Shreve, 1964). The residual radioactive contamination in the surface soil consists of weapons grade plutonium, depleted uranium, and their radioactive decay products

  4. A remote palm domain residue of RB69 DNA polymerase is critical for enzyme activity and influences the conformation of the active site.

    Directory of Open Access Journals (Sweden)

    Agata Jacewicz

    Full Text Available Non-conserved amino acids that are far removed from the active site can sometimes have an unexpected effect on enzyme catalysis. We have investigated the effects of alanine replacement of residues distant from the active site of the replicative RB69 DNA polymerase, and identified a substitution in a weakly conserved palm residue (D714A, that renders the enzyme incapable of sustaining phage replication in vivo. D714, located several angstroms away from the active site, does not contact the DNA or the incoming dNTP, and our apoenzyme and ternary crystal structures of the Pol(D714A mutant demonstrate that D714A does not affect the overall structure of the protein. The structures reveal a conformational change of several amino acid side chains, which cascade out from the site of the substitution towards the catalytic center, substantially perturbing the geometry of the active site. Consistent with these structural observations, the mutant has a significantly reduced k pol for correct incorporation. We propose that the observed structural changes underlie the severe polymerization defect and thus D714 is a remote, non-catalytic residue that is nevertheless critical for maintaining an optimal active site conformation. This represents a striking example of an action-at-a-distance interaction.

  5. In vitro mutagenesis studies at the arginine residues of adenylate kinase. A revised binding site for AMP in the X-ray-deduced model

    International Nuclear Information System (INIS)

    Kim, Hyo Joon; Nishikawa, Satoshi; Tokutomi, Yuiko; Uesugi, Seiichi; Takenaka, Hitoshi; Hamada, Minoru; Kuby, S.A.

    1990-01-01

    Although X-ray crystallographic and NMR studies have been made on the adenylate kinases, the substrate-binding sites are not unequivocally established. In an attempt to shed light on the binding sites for MgATP 2- and for AMP 2- in human cytosolic adenylate kinase, the authors have investigated the enzymic effects of replacement of the arginine residues, which had been assumed by Pai et al. to interact with the phosphoryl groups of AMP 2- and MgATP 2- . With use of the site-directed mutagenesis method, point mutations were made in the artificial gene for hAK1 to replace these arginine residues with alanyl residues and yield the mutants R44A hAK1, R132A hAK1, R138A hAK1, and R149A hAK1. The resulting large increases in the K m,app values for AMP 2- of the mutant enzymes, the relatively small increases in the K m,app values for MgATP 2- , and the fact that the R132A, R138A, and R149A mutant enzymes proved to be very poor catalysts are consistent with the idea that the assigned substrate binding sites of Pai et al. have been reversed and that their ATP-binding site may be assigned as the AMP site

  6. Distribution of 3H-fucose in the transseptal ligament of the mouse

    International Nuclear Information System (INIS)

    Johnson, R.B.

    1988-01-01

    The present study demonstrated 3 H-fucose distribution in three regions of the transseptal ligament; the middle, mesial (adjacent to the first molar tooth) and distal (adjacent to the second molar tooth) thirds; og 6-week- and 6-month-old Swiss mice. Incorporation of the isotope was rapid and maximal peaks occurred at 3 hours in most regions of 6-month and at 12 h in most regions of 6-wk animals: highest grain counts were over the middle third of the ligament of 6-wk animals (p<0.001). During incorporation, mean grain counts were significantly different as a function of age (p<0.001), region (p<0.01) and time (p<0.001) and there was a significant interaction between region and time (p<0.005). Grain removal occurred at a slower rate: there were significant differences in mean grain counts as a function of age (p<0.001), region (p<0.001) and time (p<0.001) and a significant interaction between time and region (p<0.001) and between time, region and age (p<0.001). The highest rate of grain removal was in the middle third of the ligament of 6-wk animals. Half-life of labeled glycoproteins was shortest in the ligament of 6-wk animals. The study suggested regional differences in metabolism of fucose-containing glycoproteins of the transseptal ligament: highest turnover rates in the 1) middle third, and 2) young animals. (author)

  7. In situ chemical composition measurement of individual cloud residue particles at a mountain site, southern China

    Directory of Open Access Journals (Sweden)

    Q. Lin

    2017-07-01

    Full Text Available To investigate how atmospheric aerosol particles interact with chemical composition of cloud droplets, a ground-based counterflow virtual impactor (GCVI coupled with a real-time single-particle aerosol mass spectrometer (SPAMS was used to assess the chemical composition and mixing state of individual cloud residue particles in the Nanling Mountains (1690 m a. s. l. , southern China, in January 2016. The cloud residues were classified into nine particle types: aged elemental carbon (EC, potassium-rich (K-rich, amine, dust, Pb, Fe, organic carbon (OC, sodium-rich (Na-rich and Other. The largest fraction of the total cloud residues was the aged EC type (49.3 %, followed by the K-rich type (33.9 %. Abundant aged EC cloud residues that mixed internally with inorganic salts were found in air masses from northerly polluted areas. The number fraction (NF of the K-rich cloud residues increased within southwesterly air masses from fire activities in Southeast Asia. When air masses changed from northerly polluted areas to southwesterly ocean and livestock areas, the amine particles increased from 0.2 to 15.1 % of the total cloud residues. The dust, Fe, Pb, Na-rich and OC particle types had a low contribution (0.5–4.1 % to the total cloud residues. Higher fraction of nitrate (88–89 % was found in the dust and Na-rich cloud residues relative to sulfate (41–42 % and ammonium (15–23 %. Higher intensity of nitrate was found in the cloud residues relative to the ambient particles. Compared with nonactivated particles, nitrate intensity decreased in all cloud residues except for dust type. To our knowledge, this study is the first report on in situ observation of the chemical composition and mixing state of individual cloud residue particles in China.

  8. TmiRUSite and TmiROSite scripts: searching for mRNA fragments with miRNA binding sites with encoded amino acid residues.

    Science.gov (United States)

    Berillo, Olga; Régnier, Mireille; Ivashchenko, Anatoly

    2014-01-01

    microRNAs are small RNA molecules that inhibit the translation of target genes. microRNA binding sites are located in the untranslated regions as well as in the coding domains. We describe TmiRUSite and TmiROSite scripts developed using python as tools for the extraction of nucleotide sequences for miRNA binding sites with their encoded amino acid residue sequences. The scripts allow for retrieving a set of additional sequences at left and at right from the binding site. The scripts presents all received data in table formats that are easy to analyse further. The predicted data finds utility in molecular and evolutionary biology studies. They find use in studying miRNA binding sites in animals and plants. TmiRUSite and TmiROSite scripts are available for free from authors upon request and at https: //sites.google.com/site/malaheenee/downloads for download.

  9. Long-term management of the existing radioactive wastes and residues at the Niagara Falls Storage Site. Draft Environmental Impact Statement

    International Nuclear Information System (INIS)

    1984-08-01

    The statement assesses and compares several alternatives for long-term management of the existing radioactive wastes and residues at the Niagara Falls Storage Site (NFSS), Lewiston, New York. The alternatives include: (1) no action (continued interim storage at NFSS within a diked and capped containment area), (2) long-term management at NFSS (improved containment, with or without modified form of the residues), (3) long-term management at other DOE sites (Hanford, Washington, or Oak Ridge, Tennessee), and (4) offsite management of the residues at Hanford or Oak Ridge and either leaving the wastes at NFSS or removing them for disposal in the ocean. In addition to alternatives analyzed in depth, several options are also considered, including: other modifications of residue form, modification of the basic conceptual designs, other containment design options, transportation routes, and transportation modes. The radiological health effects (primarily increased risk of cancer) associated with long-term management of the wastes and residues are expected to be smaller than the nonradiological risks of occupational and transportation-related injuries and deaths. During the action period, the risk is highest for workers if all wastes and residues are moved to Hanford. The risk is highest for the general public if the residues are moved to Hanford and the wastes are moved to the ocean. Dispersal of the slightly contaminated wastes in the ocean is not expected to result in any significant impacts on the ocean environment or pose any significant radiological risk to humans. For all alternatives, if controls ceased, there would be eventual dispersion of the radioactive materials to the environment. If it is assumed that all controls cease, predicted time for loss of covers over the buried materials ranges from several hundred years to more than two million years, depending on the use of the land surface

  10. Mutations at the S1 sites of methionine aminopeptidases from Escherichia coli and Homo sapiens reveal the residues critical for substrate specificity.

    Science.gov (United States)

    Li, Jing-Ya; Cui, Yong-Mei; Chen, Ling-Ling; Gu, Min; Li, Jia; Nan, Fa-Jun; Ye, Qi-Zhuang

    2004-05-14

    Methionine aminopeptidase (MetAP) catalyzes the removal of methionine from newly synthesized polypeptides. MetAP carries out this cleavage with high precision, and Met is the only natural amino acid residue at the N terminus that is accepted, although type I and type II MetAPs use two different sets of residues to form the hydrophobic S1 site. Characteristics of the S1 binding pocket in type I MetAP were investigated by systematic mutation of each of the seven S1 residues in Escherichia coli MetAP type I (EcMetAP1) and human MetAP type I (HsMetAP1). We found that Tyr-65 and Trp-221 in EcMetAP1, as well as the corresponding residues Phe-197 and Trp-352 in HsMetAP1, were essential for the hydrolysis of a thiopeptolide substrate, Met-S-Gly-Phe. Mutation of Phe-191 to Ala in HsMetAP1 caused inactivity in contrast to the full activity of EcMetAP1(Y62A), which may suggest a subtle difference between the two type I enzymes. The more striking finding is that mutation of Cys-70 in EcMetAP1 or Cys-202 in HsMetAP1 opens up the S1 pocket. The thiopeptolides Leu-S-Gly-Phe and Phe-S-Gly-Phe, with previously unacceptable Leu or Phe as the N-terminal residue, became efficient substrates of EcMetAP1(C70A) and HsMetAP1(C202A). The relaxed specificity shown in these S1 site mutants for the N-terminal residues was confirmed by hydrolysis of peptide substrates and inhibition by reaction products. The structural features at the enzyme active site will be useful information for designing specific MetAP inhibitors for therapeutic applications.

  11. TmiRUSite and TmiROSite scripts: searching for mRNA fragments with miRNA binding sites with encoded amino acid residues

    OpenAIRE

    Berillo, Olga; Régnier, Mireille; Ivashchenko, Anatoly

    2014-01-01

    microRNAs are small RNA molecules that inhibit the translation of target genes. microRNA binding sites are located in the untranslated regions as well as in the coding domains. We describe TmiRUSite and TmiROSite scripts developed using python as tools for the extraction of nucleotide sequences for miRNA binding sites with their encoded amino acid residue sequences. The scripts allow for retrieving a set of additional sequences at left and at right from the binding site. The scripts presents ...

  12. Bromopyruvate, an active site-directed inactivator of E. coli 2-keto-4-hydroxyglutarate(KHG) aldolase, modifies glutamic acid residue-45

    International Nuclear Information System (INIS)

    Vlahos, C.J.; Dekker, E.E.

    1987-01-01

    E. coli KHG-aldolase (2-keto-4-hydroxyglutarate ↔ pyruvate + glyoxylate), a novel trimeric Class I aldolase, requires one active-site lysine residue (Lys 133)/subunit for Schiff-base formation as well as one arginine residue (Arg 49)/subunit for catalytic activity. The substrate analog, 3-bromopyruvate (BRPY), causes a time- and concentration-dependent loss of KHG-aldolase activity. This inactivation is regarded as active site-directed since: (a) BRPY modification results in complete loss of enzymatic activity; (b) saturation kinetics are exhibited, suggesting that a reversible complex is formed between the aldolase and BRPY prior to the rate-limiting inactivation step; (c) over 90% of the initial aldolase activity is protected by either substrate, pyruvate or KHG; (d) 1.1 mol of 14 C-BRPY is bound/enzyme subunit. Peptide isolation and sequencing show that the incorporated radioactivity is associated with residue Glu-45. Denaturation of the enzyme with guanidine x HCl following treatment with excess 14 C-BRPY allows for the incorporation of carbon-14 at Cys-159 and Cys-180 as well. The presence of pyruvate protects Glu-45 from being esterified but does not prevent the alkylation of the two cysteine residues. These results suggest that Glu-45 is essential for the catalytic activity of E. coli KHG-aldolase, most likely functioning as the active-site amphoteric proton donor/acceptor moiety that is involved in the overall mechanism of the reaction catalyzed by this enzyme

  13. A simple procedure for the isolation of L-fucose-binding lectins from Ulex europaeus and Lotus tetragonolobus.

    Science.gov (United States)

    Allen, H J; Johnson, E A

    1977-10-01

    L-Fucose-binding lectins from Ulex europeaus and Lotus tetragonolobus were isolated by affinity chromatography on columns of L-fucose-Sepharose 6B. L-Fucose was coupled to Sepharose 6B after divinyl sulfone-activation of the gel to give an affinity adsorbent capable of binding more than 1.2 mg of Ulex lextin/ml of gel, which could then be eluted with 0.1M or 0.05M L-fucose. Analysis of the isolated lectins by hemagglutination assay, by gel filtration, and polyacrylamide disc-electrophoresis revealed the presence of isolectins, or aggregated species, or both. The apparent mol. wt. of the major lectin fraction from Lotus was 35000 when determined on Sephadex G-200 or Ultrogel AcA 34. In contrast, the apparent mol. wt. of the major lectin fraction from Ulex was 68 000 when chromatographed on Sephadex G-200 and 45 000 when chromatographed on Ultrogel AcA 34. The yields of lectins were 4.5 mg/100 g of Ulex seeds and 394 mg/100 g of Lotus seeds.

  14. Comparison of the antimicrobial adhesion potential of human body fluid glycoconjugates using fucose-binding lectin (PA-IIL) of Pseudomonas aeruginosa and Ulex europaeus lectin (UEA-I).

    Science.gov (United States)

    Lerrer, Batia; Lesman-Movshovich, Efrat; Gilboa-Garber, Nechama

    2005-09-01

    Pseudomonas aeruginosa produces a fucose-binding lectin (PA-IIL) which strongly binds to human cells. This lectin was shown to be highly sensitive to inhibition by fucose-bearing human milk glycoproteins. Since the glycans of these glycoproteins mimic human cell receptors, they may function as decoys in blocking lectin-dependent pathogen adhesion to the host cells. Human saliva and seminal fluid also contain such compounds, and body fluids of individuals who are "secretors" express additional fucosylated (alpha 1,2) residues. The latter are selectively detected by Ulex europaeus lectin UEA-I. The aim of the present research was to compare the PA-IIL and UEA-I interactions with human salivas and seminal fluids of "secretors" and "nonsecretors" with those obtained with the respective milks. Using hemagglutination inhibition and Western blot analyses, we showed that PA-IIL interactions with the saliva and seminal fluid glycoproteins were somewhat weaker than those obtained with the milk and that "nonsecretor" body fluids were not less efficient than those of "secretors" in PA-IIL blocking. UEA-I, which interacted only with the "secretors" glycoproteins, was most sensitive to those of the seminal fluids.

  15. Strategy utilized for assessing baseline risks to human health from K-65 and metal oxide residues stored at the Fernald Site

    International Nuclear Information System (INIS)

    Harmon, J.E.; Janke, R.C.

    1995-01-01

    The U.S. Department of Energy (DOE) is responsible for cleanup activities at the Fernald Environmental Management Project (FEMP) site in southwestern Ohio. The 425-hectare site consists of a former 55-hectare Production Area, an adjacent Waste Storage Area and various support facilities. From 1952 until 1989, the FEMP processed uranium into metallic open-quotes feedclose quotes materials for other DOE facilities in the nation's defense program. In accordance with the Comprehensive Environmental Response, Compensation and Liability Act (CERCLA), the FEMP site is currently listed on the National Priorities List (NPL). To facilitate an expeditious cleanup effort, environmental issues associated with site cleanup are being managed under five operable units. This paper summarizes the risk assessment strategy employed to determine baseline human health risks associated with K-65 and metal oxide residues currently stored in Operable Unit 4. The K-65 and metal oxide residues were generated during the 1950s as a result of the extraction of uranium from uranium-bearing ores and concentrates. These residues are currently stored within Operable Unit 4 in concrete silos. Silos I and 2 contain approximately 6,120 cubic meters [m 3 ] (8,005 cubic yards [yd 3 ]) of K-65 residues, while silos 3 contains approximately 3890 m 3 (5,080 yd 3 ) of cold metal oxides. These concrete silos are beyond their design life and require remedial action. The risk assessment conducted for Operable Unit 4 constitutes the first detailed human health risk assessment to be approved by the Environmental Protection Agency (EPA) for the CERCLA clean-up effort at the FEMP Site. This paper discusses the FEMP's use of a Risk Information Quality Objective process in concert with the traditional risk assessment approach to determine baseline risk to human health and the environment posed by Operable Unit 4. A summary of the baseline risks to human health is also presented

  16. Strategy utilized for assessing baseline risks to human health from K-65 and metal oxide residues stored at the Fernald Site

    Energy Technology Data Exchange (ETDEWEB)

    Harmon, J.E. [FERMCO, Cincinnati, OH (United States). Fernald Environmental Management Project; Janke, R.C.

    1995-04-01

    The U.S. Department of Energy (DOE) is responsible for cleanup activities at the Fernald Environmental Management Project (FEMP) site in southwestern Ohio. The 425-hectare site consists of a former 55-hectare Production Area, an adjacent Waste Storage Area and various support facilities. From 1952 until 1989, the FEMP processed uranium into metallic {open_quotes}feed{close_quotes} materials for other DOE facilities in the nation`s defense program. In accordance with the Comprehensive Environmental Response, Compensation and Liability Act (CERCLA), the FEMP site is currently listed on the National Priorities List (NPL). To facilitate an expeditious cleanup effort, environmental issues associated with site cleanup are being managed under five operable units. This paper summarizes the risk assessment strategy employed to determine baseline human health risks associated with K-65 and metal oxide residues currently stored in Operable Unit 4. The K-65 and metal oxide residues were generated during the 1950s as a result of the extraction of uranium from uranium-bearing ores and concentrates. These residues are currently stored within Operable Unit 4 in concrete silos. Silos I and 2 contain approximately 6,120 cubic meters [m{sup 3}] (8,005 cubic yards [yd{sup 3}]) of K-65 residues, while silos 3 contains approximately 3890 m{sup 3} (5,080 yd{sup 3}) of cold metal oxides. These concrete silos are beyond their design life and require remedial action. The risk assessment conducted for Operable Unit 4 constitutes the first detailed human health risk assessment to be approved by the Environmental Protection Agency (EPA) for the CERCLA clean-up effort at the FEMP Site. This paper discusses the FEMP`s use of a Risk Information Quality Objective process in concert with the traditional risk assessment approach to determine baseline risk to human health and the environment posed by Operable Unit 4. A summary of the baseline risks to human health is also presented.

  17. Influence of temperature on the rheological behavior of a new fucose-containing bacterial exopolysaccharide.

    Science.gov (United States)

    Cruz, Madalena; Freitas, Filomena; Torres, Cristiana A V; Reis, Maria A M; Alves, Vítor D

    2011-05-01

    The effect of temperature on the rheology of a new fucose-containing extracellular polysaccharide (EPS) was evaluated. The steady state data revealed a shear-thinning behavior, with the viscosity being immediately recovered when the shear rate was decreased. The mechanical spectra indicated viscous solutions with entangled polymer molecules in the range of temperatures studied (from 15 °C to 65 °C). In addition, the Time-Temperature Superposition principle was successfully applied and the Cox-Merz rule was valid, reinforcing the idea of a thermorheologically simple behavior for the EPS in aqueous solution. Furthermore, the viscous and viscoelastic properties at 25 °C were maintained after consecutive heating and cooling cycles, indicating a good thermal stability under temperature fluctuations. Copyright © 2011 Elsevier B.V. All rights reserved.

  18. L-Fucose as a terminal sugar in cellular glycoconjugates of colonic carcinoma

    Directory of Open Access Journals (Sweden)

    Sargazei GH

    2008-10-01

    Full Text Available "n Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 st1":*{behavior:url(#ieooui } /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Arial; mso-bidi-theme-font:minor-bidi;} Background: Glycoconjugates are a class of cell surface glycoproteins, the terminal sugars of which are important indicators of neoplasia and the aberrant biological behavior of cancer cells. Lectins are a class of plant or animal glycoproteins that specifically bind to the terminal sugars of glycoconjugates. The aim of the present study is to identify the presence of L-fucose in cell surface glycoconjugates and extracellular matrix glycoconjugates of cancer cells of different grades of colonic adenocarcinoma."n"n Methods: Paraffin blocks of colonic adenocarcinoma tissue from 30 patients were collected from the Pathology Department of Khatam Al Anbia Hospital in Zahedan, Iran. Sections, 5-7μm thick, were prepared and stained using hematoxylin and eosin. Sections were graded histopathologically and then stained using the lectin Ulex europaeus agglutinin (UEA, 10μm/mL, which binds specifically to L-fucose, and Alcian blue (pH=2.5. Sections were graded blindly according to lectin staining intensity on a scale of 0-3. Collected data were analyzed using Kruskall-Wallis and Mann Whitney nonparametric tests with SPSS."n"n Results: Our results show that there is a significant difference in the

  19. Conserved Residues Lys57 and Lys401 of Protein Disulfide Isomerase Maintain an Active Site Conformation for Optimal Activity: Implications for Post-Translational Regulation

    Directory of Open Access Journals (Sweden)

    Cody Caba

    2018-02-01

    Full Text Available Despite its study since the 1960's, very little is known about the post-translational regulation of the multiple catalytic activities performed by protein disulfide isomerase (PDI, the primary protein folding catalyst of the cell. This work identifies a functional role for the highly conserved CxxC-flanking residues Lys57 and Lys401 of human PDI in vitro. Mutagenesis studies have revealed these residues as modulating the oxidoreductase activity of PDI in a pH-dependent manner. Non-conservative amino acid substitutions resulted in enzyme variants upwards of 7-fold less efficient. This attenuated activity was found to translate into a 2-fold reduction of the rate of electron shuttling between PDI and the intraluminal endoplasmic reticulum oxidase, ERO1α, suggesting a functional significance to oxidative protein folding. In light of this, the possibility of lysine acetylation at residues Lys57 and Lys401 was assessed by in vitro treatment using acetylsalicylic acid (aspirin. A total of 28 acetyllysine residues were identified, including acLys57 and acLys401. The kinetic behavior of the acetylated protein form nearly mimicked that obtained with a K57/401Q double substitution variant providing an indication that acetylation of the active site-flanking lysine residues can act to reversibly modulate PDI activity.

  20. Conserved Residues Lys57 and Lys401 of Protein Disulfide Isomerase Maintain an Active Site Conformation for Optimal Activity: Implications for Post-Translational Regulation.

    Science.gov (United States)

    Caba, Cody; Ali Khan, Hyder; Auld, Janeen; Ushioda, Ryo; Araki, Kazutaka; Nagata, Kazuhiro; Mutus, Bulent

    2018-01-01

    Despite its study since the 1960's, very little is known about the post-translational regulation of the multiple catalytic activities performed by protein disulfide isomerase (PDI), the primary protein folding catalyst of the cell. This work identifies a functional role for the highly conserved CxxC-flanking residues Lys 57 and Lys 401 of human PDI in vitro . Mutagenesis studies have revealed these residues as modulating the oxidoreductase activity of PDI in a pH-dependent manner. Non-conservative amino acid substitutions resulted in enzyme variants upwards of 7-fold less efficient. This attenuated activity was found to translate into a 2-fold reduction of the rate of electron shuttling between PDI and the intraluminal endoplasmic reticulum oxidase, ERO1α, suggesting a functional significance to oxidative protein folding. In light of this, the possibility of lysine acetylation at residues Lys 57 and Lys 401 was assessed by in vitro treatment using acetylsalicylic acid (aspirin). A total of 28 acetyllysine residues were identified, including acLys 57 and acLys 401 . The kinetic behavior of the acetylated protein form nearly mimicked that obtained with a K57/401Q double substitution variant providing an indication that acetylation of the active site-flanking lysine residues can act to reversibly modulate PDI activity.

  1. Influence of amino acid residues near the active site of cytochrome P450 from Bacillus megaterium on the selectivity of n-octane oxidation to octanol regioisomers

    Science.gov (United States)

    Miyaji, Akimitsu; Baba, Toshihide

    2017-09-01

    A mutant of cytochrome P450 from Bacillus megaterium (CYP450BM-3) was prepared by replacing two alanine residues around active site of the enzyme, alanine 328 and alanine 82, with leucine and tryptophan, respectively. The CYP450BM-3 mutant produced 2-octanol selectively from n-octane under atmospheric temperature and pressure; its selectivity was 74%. Furthermore, the mutant produced 1-octanol, which is not produced by wild-type enzyme.

  2. Analysis of the technical capabilities of DOE sites for disposal of residuals from the treatment of mixed low-level waste

    International Nuclear Information System (INIS)

    Waters, R.D.; Gruebel, M.M.; Langkopf, B.S.; Kuehne, P.B.

    1997-04-01

    The US Department of Energy (DOE) has stored or expects to generate over the next five years more than 130,000 m 3 of mixed low-level waste (MLLW). Before disposal, MLLW is usually treated to comply with the land disposal restrictions of the Resource Conservation and Recovery Act. Depending on the type of treatment, the original volume of MLLW and the radionuclide concentrations in the waste streams may change. These changes must be taken into account in determining the necessary disposal capacity at a site. Treatment may remove the characteristic in some waste that caused it to be classified as mixed. Treatment of some waste may, by reduction of the mass, increase the concentrations of some transuranic radionuclides sufficiently so that it becomes transuranic waste. In this report, the DOE MLLW streams were analyzed to determine after-treatment volumes and radionuclide concentrations. The waste streams were reclassified as residual MLLW or low-level or transuranic waste resulting from treatment. The volume analysis indicated that about 89,000 m 3 of waste will require disposal as residual MLLW. Fifteen DOE sites were then evaluated to determine their capabilities for hosting disposal facilities for some or all of the residual MLLW. Waste streams associated with about 90% of the total residual MLLW volume are likely to present no significant issues for disposal and require little additional analysis. Future studies should focus on the remaining waste streams that are potentially problematic by examining site-specific waste acceptance criteria, alternative treatment processes, alternative waste forms for disposal, and pending changes in regulatory requirements

  3. Magnesium-adenosine diphosphate binding sites in wild-type creatine kinase and in mutants: role of aromatic residues probed by Raman and infrared spectroscopies.

    Science.gov (United States)

    Hagemann, H; Marcillat, O; Buchet, R; Vial, C

    2000-08-08

    Two distinct methods were used to investigate the role of Trp residues during Mg-ADP binding to cytosolic creatine kinase (CK) from rabbit muscle: (1) Raman spectroscopy, which is very sensitive to the environment of aromatic side-chain residues, and (2) reaction-induced infrared difference spectroscopy (RIDS) and photolabile substrate (ADP[Et(PhNO(2))]), combined with site-directed mutagenesis on the four Trp residues of CK. Our Raman results indicated that the environment of Trp and of Tyr were not affected during Mg-ADP binding to CK. Analysis of RIDS of wild-type CK, inactive W227Y, and active W210,217,272Y mutants suggested that Trp227 was not involved in the stacking interactions. Results are consistent with Trp227 being essential to prevent water molecules from entering in the active site [as suggested by Gross, M., Furter-Graves, E. M., Wallimann, T., Eppenberger, H. M., and Furter, R. (1994) Protein Sci. 3, 1058-1068] and that another Trp could in addition help to steer the nucleotide in the binding site, although it is not essential for the activity of CK. Raman and infrared spectra indicated that Mg-ADP binding does not involve large secondary structure changes. Only 3-4 residues absorbing in the amide I region are directly implicated in the Mg-ADP binding (corresponding to secondary structure changes less than 1%), suggesting that movement of protein domains due to Mg-nucleotide binding do not promote large secondary structure changes.

  4. Influenza human monoclonal antibody 1F1 interacts with three major antigenic sites and residues mediating human receptor specificity in H1N1 viruses.

    Directory of Open Access Journals (Sweden)

    Tshidi Tsibane

    Full Text Available Most monoclonal antibodies (mAbs to the influenza A virus hemagglutinin (HA head domain exhibit very limited breadth of inhibitory activity due to antigenic drift in field strains. However, mAb 1F1, isolated from a 1918 influenza pandemic survivor, inhibits select human H1 viruses (1918, 1943, 1947, and 1977 isolates. The crystal structure of 1F1 in complex with the 1918 HA shows that 1F1 contacts residues that are classically defined as belonging to three distinct antigenic sites, Sa, Sb and Ca(2. The 1F1 heavy chain also reaches into the receptor binding site (RBS and interacts with residues that contact sialoglycan receptors and determine HA receptor specificity. The 1F1 epitope is remarkably similar to the previously described murine HC63 H3 epitope, despite significant sequence differences between H1 and H3 HAs. Both antibodies potently inhibit receptor binding, but only HC63 can block the pH-induced conformational changes in HA that drive membrane fusion. Contacts within the RBS suggested that 1F1 may be sensitive to changes that alter HA receptor binding activity. Affinity assays confirmed that sequence changes that switch the HA to avian receptor specificity affect binding of 1F1 and a mAb possessing a closely related heavy chain, 1I20. To characterize 1F1 cross-reactivity, additional escape mutant selection and site-directed mutagenesis were performed. Residues 190 and 227 in the 1F1 epitope were found to be critical for 1F1 reactivity towards 1918, 1943 and 1977 HAs, as well as for 1I20 reactivity towards the 1918 HA. Therefore, 1F1 heavy-chain interactions with conserved RBS residues likely contribute to its ability to inhibit divergent HAs.

  5. Newly identified essential amino acid residues affecting ^8-sphingolipid desaturase activity revealed by site-directed mutagenesis

    Science.gov (United States)

    In order to identify amino acid residues crucial for the enzymatic activity of ^8-sphingolipid desaturases, a sequence comparison was performed among ^8-sphingolipid desaturases and ^6-fatty acid desaturase from various plants. In addition to the known conserved cytb5 (cytochrome b5) HPGG motif and...

  6. Salmonella enterica serotype Typhimurium Std fimbriae bind terminal α (1,2)fucose residues in the cecal mucosa

    Science.gov (United States)

    Chessa, Daniela; Winter, Maria G.; Jakomin, Marcello; Bäumler, Andreas J.

    2013-01-01

    SUMMARY The std operon encodes a fimbrial adhesin of Salmonella enterica serotype Typhimurium that is required for attachment to intestinal epithelial cells and for cecal colonization in the mouse. To study the mechanism by which this virulence factor contributes to colonization we characterized its binding specificity. Std-mediated binding to human colonic epithelial (Caco-2) cells could be abrogated by removing N-linked glycans. Adherence of Std fimbriated S. Typhimurium to Caco-2 cells could be blocked by co-incubation with H type 2 oligosaccharide (Fucα1-2Galβ1-4GlcNAc) or by pretreatment of cells with α1-2 fucosidase. In contrast, pretreatment of Caco-2 cells with neuraminidase or co-incubation with the type 2 disaccharide precursor (Galβ1-4GlcNAc) did not reduce adherence of Std fimbriated S. Typhimurium. Binding of purified Std fimbriae to Fucα1-2Galβ1-4GlcNAc in a solid phase binding assay was competitively inhibited by Ulex europaeus agglutinin-I (UEA-I), a lectin specific for Fucα1-2 moieties. Purified Std fimbriae and UEA both bound to a receptor localized in the mucus layer of the murine cecum. These data suggest that the std operon encodes an adhesin that binds an α1-2 fucosylated receptor(s) present in the cecal mucosa. PMID:19183274

  7. Mass spectrometry and site-directed mutagenesis identify several autophosphorylated residues required for the activity of PrkC, a Ser/Thr kinase from Bacillus subtilis

    DEFF Research Database (Denmark)

    Madec, Edwige; Stensballe, Allan; Kjellström, Sven

    2003-01-01

    We have shown recently that PrkC, which is involved in developmental processes in Bacillus subtilis, is a Ser/Thr kinase with features of the receptor kinase family of eukaryotic Hanks kinases. In this study, we expressed and purified from Escherichia coli the cytoplasmic domain of PrkC containing...... the kinase and a short juxtamembrane region. This fragment, which we designate PrkCc, undergoes autophosphorylation in E.coli. PrkCc is further autophosphorylated in vitro, apparently through a trans-kinase, intermolecular reaction. PrkC also displays kinase activity with myelin basic protein. Using high...... mass accuracy electrospray tandem mass spectrometry (LC-MS/MS) and nanoelectrospray tandem mass spectrometry, we identified seven phosphorylated threonine and one serine residue in PrkCc. All the corresponding residues were replaced by systematic site-directed mutagenesis and the purified mutant...

  8. Site-Mutation of Hydrophobic Core Residues Synchronically Poise Super Interleukin 2 for Signaling: Identifying Distant Structural Effects through Affordable Computations

    Directory of Open Access Journals (Sweden)

    Longcan Mei

    2018-03-01

    Full Text Available A superkine variant of interleukin-2 with six site mutations away from the binding interface developed from the yeast display technique has been previously characterized as undergoing a distal structure alteration which is responsible for its super-potency and provides an elegant case study with which to get insight about how to utilize allosteric effect to achieve desirable protein functions. By examining the dynamic network and the allosteric pathways related to those mutated residues using various computational approaches, we found that nanosecond time scale all-atom molecular dynamics simulations can identify the dynamic network as efficient as an ensemble algorithm. The differentiated pathways for the six core residues form a dynamic network that outlines the area of structure alteration. The results offer potentials of using affordable computing power to predict allosteric structure of mutants in knowledge-based mutagenesis.

  9. Reduction of N-linked xylose and fucose by expression of rat beta1,4-N-acetylglucosaminyltransferase III in tobacco BY-2 cells depends on Golgi enzyme localization domain and genetic elements used for expression.

    Science.gov (United States)

    Karg, Saskia R; Frey, Alexander D; Kallio, Pauli T

    2010-03-01

    Plant-specific N-glycosylation, such as the introduction of core alpha1,3-fucose and beta1,2-xylose residues, is a major obstacle to the utilization of plant cell- or plant-derived recombinant therapeutic proteins. The beta1,4-N-acetylglucosaminyltransferase III (GnTIII) introduces a bisecting GlcNAc residue into N-glycans, which exerts a high level of substrate mediated control over subsequent modifications, for example inhibiting mammalian core fucosylation. Based on similar findings in plants, we used Nicotianatabacum BY-2 cells to study the effects of localization and expression levels of GnTIII in the remodeling of the plant N-glycosylation pathway. The N-glycans produced by the cells expressing GnTIII were partially bisected and practically devoid of the paucimannosidic type which is typical for N-glycans produced by wildtype BY-2 suspension cultured cells. The proportion of human-compatible N-glycans devoid of fucose and xylose could be increased from an average of 4% on secreted protein from wildtype cells to as high as 59% in cells expressing chimeric GnTIII, named GnTIII(A.th.) replacing its native localization domain with the cytoplasmic tail, transmembrane, and stem region of Arabidopsis thaliana mannosidase II. The changes in N-glycosylation observed were dependent on the catalytic activity of GnTIII, as the expression of catalytically inactive GnTIII mutants did not show a significant effect on N-glycosylation. Copyright 2010 Elsevier B.V. All rights reserved.

  10. Structural and metabolic studies of O-linked fucose-containing proteins of normal and virally-transformed rat fibroblasts

    International Nuclear Information System (INIS)

    Morton, P.A.

    1985-01-01

    Previous studies in this laboratory have demonstrated that cultured human and rodent cells contain a series of low molecular weight glycosylated amino acids of unusual structure, designated amino acid fucosides. The incorporation of radiolabelled-fucose into one of these components, designated FL4a (glucosylfucosylthreonine), is markedly-reduced in transformed epithelial and fibroblastic cells. The authors have examined fucose-labelled normal and virally-transformed rat fibroblast cell lines for glycoproteins which might be precursors to amino acid fucosides. Using milk alkaline/borohydride treatment (the beta-elimination reaction) to release O-linked oligosaccharides from proteins, they have isolated and partially characterized two low M/sub r/ reaction products (designated DS-ol and TS-ol) released from macromolecular cell material. The identity of one of these components (DS-ol, glucosylfucitol) suggested the existence in these cells of a direct protein precursor to FL4a. They examined fucose-labelled macromolecular cell material for proteins which release DS-ol (DS-proteins.). Using gel filtration chromatography and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) with subsequent autoradiography, they have observed DS-proteins which appear to exhibit a broad molecular weight size range, and are also present in culture medium from normal and transformed cells. The findings suggest that mammalian cells contain DS-proteins and TS-proteins with a novel carbohydrate-peptide linkage wherein L-fucose is O-linked to a polypeptide backbone. Metabolic studies were undertaken to examine both the relationship between DS-protein and FL4a and the biochemical basis for the decreased level of FL4a and the biochemical basis for the decreased level of FL4a observed in transformed cells

  11. Involved Node, Site, Field and Residual Volume Radiotherapy for Lymphoma: A Comparison of Organ at Risk Dosimetry and Second Malignancy Risks.

    Science.gov (United States)

    Murray, L; Sethugavalar, B; Robertshaw, H; Bayman, E; Thomas, E; Gilson, D; Prestwich, R J D

    2015-07-01

    Recent radiotherapy guidelines for lymphoma have included involved site radiotherapy (ISRT), involved node radiotherapy (INRT) and irradiation of residual volume after full-course chemotherapy. In the absence of late toxicity data, we aim to compare organ at risk (OAR) dose-metrics and calculated second malignancy risks. Fifteen consecutive patients who had received mediastinal radiotherapy were included. Four radiotherapy plans were generated for each patient using a parallel pair photon technique: (i) involved field radiotherapy (IFRT), (ii) ISRT, (iii) INRT, (iv) residual post-chemotherapy volume. The radiotherapy dose was 30 Gy in 15 fractions. The OARs evaluated were: breasts, lungs, thyroid, heart, oesophagus. Relative and absolute second malignancy rates were estimated using the concept of organ equivalent dose. Significance was defined as P risks of second cancers were significantly higher with IFRT compared with ISRT for lung, breast and thyroid; INRT and residual volume resulted in significantly lower relative risks compared with ISRT for lung, breast and thyroid. The median excess absolute risks of second cancers were consistently lowest for the residual technique and highest for IFRT in terms of thyroid, lung and breast cancers. The risk of oesophageal cancer was similar for all four techniques. Overall, the absolute risk of second cancers was very similar for ISRT and INRT. Decreasing treatment volumes from IFRT to ISRT, INRT or residual volume reduces radiation exposure to OARs. Second malignancy modelling suggests that this reduction in treatment volumes will lead to a reduction in absolute excess second malignancy. Little difference was observed in second malignancy risks between ISRT and INRT, supporting the use of ISRT in the absence of a pre-chemotherapy positron emission tomography scan in the radiotherapy treatment position. Copyright © 2015 The Royal College of Radiologists. Published by Elsevier Ltd. All rights reserved.

  12. Work plan for the remedial investigation/feasibility study-environmental assessment for the quarry residuals operable unit at the Weldon Spring Site

    International Nuclear Information System (INIS)

    1994-01-01

    The US Department of Energy (DOE) is conducting cleanup activities at the Weldon Spring site, which is located in St. Charles County, Missouri, about 48 km (30 mi) west of St. Louis. The Weldon Spring site consists of two noncontiguous areas -- the chemical plant area, which includes four raffinate pits, and the quarry. Cleanup activities at the Weldon Spring site are conducted in accordance with the Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA), as amended, incorporating the values of the National Environmental Policy Act (NEPA). The contents of the documents prepared for the project are not intended to represent a statement regarding the legal applicability of NEPA to remedial actions conducted under CERCLA. In accordance with the integrated CERCLA/NEPA approach, a remedial investigation/feasibility study-environmental assessment (RI/FS-EA) is being conducted to evaluate conditions and potential responses for the quarry residuals operable unit (QROU). This operable unit consists of the following areas and/or media: the residual material remaining at the Weldon Spring quarry after removal of the pond water and bulk waste; underlying groundwater; and other media located in the surrounding vicinity of the quarry, including adjacent soil, surface water, and sediment in Femme Osage Slough. This work plan identifies the activities within the RI/FS-EA process that are being proposed to address contamination remaining at the quarry area

  13. Work plan for the remedial investigation/feasibility study-environmental assessment for the quarry residuals operable unit at the Weldon Spring Site

    Energy Technology Data Exchange (ETDEWEB)

    1994-01-01

    The US Department of Energy (DOE) is conducting cleanup activities at the Weldon Spring site, which is located in St. Charles County, Missouri, about 48 km (30 mi) west of St. Louis. The Weldon Spring site consists of two noncontiguous areas -- the chemical plant area, which includes four raffinate pits, and the quarry. Cleanup activities at the Weldon Spring site are conducted in accordance with the Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA), as amended, incorporating the values of the National Environmental Policy Act (NEPA). The contents of the documents prepared for the project are not intended to represent a statement regarding the legal applicability of NEPA to remedial actions conducted under CERCLA. In accordance with the integrated CERCLA/NEPA approach, a remedial investigation/feasibility study-environmental assessment (RI/FS-EA) is being conducted to evaluate conditions and potential responses for the quarry residuals operable unit (QROU). This operable unit consists of the following areas and/or media: the residual material remaining at the Weldon Spring quarry after removal of the pond water and bulk waste; underlying groundwater; and other media located in the surrounding vicinity of the quarry, including adjacent soil, surface water, and sediment in Femme Osage Slough. This work plan identifies the activities within the RI/FS-EA process that are being proposed to address contamination remaining at the quarry area.

  14. Chemical-modification studies of a unique sialic acid-binding lectin from the snail Achatina fulica. Involvement of tryptophan and histidine residues in biological activity.

    Science.gov (United States)

    Basu, S; Mandal, C; Allen, A K

    1988-01-01

    A unique sialic acid-binding lectin, achatininH (ATNH) was purified in single step from the haemolymph of the snail Achatina fulica by affinity chromatography on sheep submaxillary-gland mucin coupled to Sepharose 4B. The homogeneity was checked by alkaline gel electrophoresis, immunodiffusion and immunoelectrophoresis. Amino acid analysis showed that the lectin has a fairly high content of acidic amino acid residues (22% of the total). About 1.3% of the residues are half-cystine. The glycoprotein contains 21% carbohydrate. The unusually high content of xylose (6%) and fucose (2.7%) in this snail lectin is quite interesting. The protein was subjected to various chemical modifications in order to detect the amino acid residues and carbohydrate residues present in its binding sites. Modification of tyrosine and arginine residues did not affect the binding activity of ATNH; however, modification of tryptophan and histidine residues led to a complete loss of its biological activity. A marked decrease in the fluorescence emission was found as the tryptophan residues of ATNH were modified. The c.d. data showed the presence of an identical type of conformation in the native and modified agglutinin. The modification of lysine and carboxy residues partially diminished the biological activity. The activity was completely lost after a beta-elimination reaction, indicating that the sugars are O-glycosidically linked to the glycoprotein's protein moiety. This result confirms that the carbohydrate moiety also plays an important role in the agglutination property of this lectin. Images Fig. 3. PMID:3140796

  15. [Identification of anopheles breeding sites in the residual foci of low malaria transmission «hotspots» in Central and Western Senegal].

    Science.gov (United States)

    Sy, O; Konaté, L; Ndiaye, A; Dia, I; Diallo, A; Taïrou, F; Bâ, E L; Gomis, J F; Ndiaye, J L; Cissé, B; Gaye, O; Faye, O

    2016-02-01

    Malaria incidence has markedly declined in the Mbour, Fatick, Niakhar and Bambey districts (central and western Senegal) thanks to a scaling up of effective control measures namely LLINs (Long Lasting Insecticide Treated Net), ACTs (Artesunate Combination Therapy) and promoting care seeking. However malaria cases are now maintained by foci of transmission called hotspots. We evaluate the role of anopheles breeding sites in the identification of malaria hotspots in the health districts of Mbour, Fatick, Niakhar and Bambey. Surveys of breeding sites were made in 6 hotspot villages and 4 non-hotspot villages. A sample was taken in each water point with mosquito larvae by dipping method and the collected specimens were identified to the genus level. Additional parameters as name of the village and breeding sites, type of collection, original water turbidity, presence of vegetation, proximity to dwellings, geographic coordinates, sizes were also collected. Sixty-two water collections were surveyed and monitored between 2013 and 2014. Temporary natural breeding sites were predominant regardless of the epidemiological status of the village. Among the 31 breeding sites located within 500 meters of dwellings in hotspots villages, 70% carried Anopheles larvae during the rainy season while 43% of the 21 breeding sites located at similar distances in non-hotspot villages carried Anopheles larvae during the same period (P = 0.042). At the end of the rainy season, the trend is the same with 27% of positive breeding sites in hotspots and 14% in non-hotspots villages. The breeding sites encountered in hotspots villages are mostly small to medium size and are more productive by Anopheles larvae than those found in non-hotspot area. This study showed that the high frequency of smallest and productive breeding sites around and inside the villages can create conditions of residual transmission.

  16. Catalytically important amino-acid residues of abalone alginate lyase HdAly assessed by site-directed mutagenesis

    OpenAIRE

    Yamamoto, Sayo; Sahara, Takehiko; Sato, Daisuke; Kawasaki, Kosei; Ohgiya, Satoru; Inoue, Akira; Ojima, Takao

    2008-01-01

    Alginate lyase is an enzyme that degrades alginate chains via β-elimination and has been used for the production of alginate oligosaccharides and protoplasts from brown algae. Previously, we deduced the amino-acid sequence of an abalone alginate lyase, HdAly, from its cDNA sequence and, through multiple amino-acid sequence alignment, found that several basic amino-acid residues were highly conserved among the polysaccharide-lyase family 14 (PL-14) enzymes including HdAly. In the present study...

  17. 1,3,4-Tri-O-acetyl-2-N-(trifluoroacetyl-β-l-fucose

    Directory of Open Access Journals (Sweden)

    David C. McCutcheon

    2014-02-01

    Full Text Available The title compound, C14H18F3NO8, was produced through conjugation of 1,3,4-tri-O-acetyl-2-azidodeoxy-α,β-l-fucose with trifluoroacetyl chloride in the presence of bis(diphenylphosphinoethane in tetrahydrofuran at room temperature. The X-ray crystal structure reveals that the β-anomer of the product mixture crystallizes from ethyl acetate/hexanes. The compound exists in a typical chair conformation with the maximum possible number of substituents, four out of five, located in the sterically preferred equatorial positions. The major directional force facilitating packing of the molecules are N—H...O hydrogen bonds involving the amide moieties of neighboring molecules, which connect molecules stacked along the a-axis direction into infinite strands with a C11(4 graph-set motif. Formation of the strands is assisted by a number of weaker C—H...O interactions involving the methine and methyl H atoms. These strands are connected through further C—H...O and C—H...F interactions into a three dimensional network

  18. Biodegradation of Aged Residues of Atrazine and Alachlor in a Mix-Load Site Soil by Fungal Enzymes

    OpenAIRE

    Chirnside, Anastasia E. M.; Ritter, William F.; Radosevich, Mark

    2011-01-01

    Soils from bulk pesticide mixing and loading (mix-load) sites are often contaminated with a complex mixture of pesticides, herbicides, and other organic compounds used in pesticide formulations that limits the success of remediation efforts. Therefore, there is a need to find remediation strategies that can successfully clean up these mix-load site soils. This paper examined the degradation of atrazine (2-chloro-4-ethylamino-6-isopropylamino-S-triazine; AT) and alachlor (2-chloro- 2  , 6  -...

  19. An efficient method to control high mannose and core fucose levels in glycosylated antibody production using deoxymannojirimycin.

    Science.gov (United States)

    Shalel Levanon, Sagit; Aharonovitz, Orit; Maor-Shoshani, Ayelet; Abraham, Gita; Kenett, Dan; Aloni, Yehoshua

    2018-06-20

    Glycosylation on the Fc region of recombinant Immunoglobulin G (IgG) therapeutic antibodies is a critical protein quality attribute which may affect the efficacy and safety of the molecule. During the development of biosimilar therapeutics, adjustment of the glycosylation profile is required in order to match the reference innovator profile. Deoxymannojirimycin (DMJ), a known inhibitor of mannosidase, was used in this study to modulate the glycosylation pattern of antibodies. The effect of DMJ, at concentrations of 5 μM - 500 μM, on non-fucosylated glycoform levels was tested in the biosynthesis processes of two different IgG1 (IgG1 #A and IgG1 #B) using two Chinese hamster ovary (CHO) cell lines (CHO-DXB-11 and CHOK1SV, respectively) in Erlenmeyer flasks and in lab scale bioreactors. DMJ affected glycan forms in a dose response manner. At the highest concentration tested, DMJ reduced N-linked complex glycoform and core fucose levels by 15 and 14 fold, respectively, and increased high mannose level by 21 fold. 10 μM DMJ decreased IgG1 #A core fucose level in CHO-DXB-11 from 92% to 73% and increased high mannose level from 4% to 22% in Erlenmeyer flasks. Furthermore, in lab scale bioreactors, 15 μM DMJ decreased IgG1 #A core fucose level from 95% to 84% and increased high mannose level from 3% to 13%. Core fucose level of IgG1 #B in CHOK1SV was decreased from 81% to 73% using 10 μM DMJ in lab scale bioreactors while high mannose was increased from 6% to 15%. While affecting core fucose and high mannose levels, DMJ decreased maximum viable cell concentration by 16% and did not significantly affect cell productivity (less than 10%). This study demonstrated that DMJ can enable the control of core fucosylated and high mannose levels of IgG1 antibodies in a defined range. Copyright © 2018 Elsevier B.V. All rights reserved.

  20. cDNA cloning of porcine brain prolyl endopeptidase and identification of the active-site seryl residue

    Energy Technology Data Exchange (ETDEWEB)

    Rennex, D.; Hemmings, B.A.; Hofsteenge, J.; Stone, S.R. (Friedrich Miescher-Institut, Basel (Switzerland))

    1991-02-26

    Prolyl endopeptidase is a cytoplasmic serine protease. The enzyme was purified from porcine kidney, and oligonucleotides based on peptide sequences from this protein were used to isolate a cDNA clone from a porcine brain library. This clone contained the complete coding sequence of prolyl endopeptidase and encoded a polypeptide with a molecular mass of 80751 Da. The deduced amino acid sequence of prolyl endopeptidase showed no sequence homology with other known serine proteases. ({sup 3}H)Diisopropyl fluorophosphate was used to identify the active-site serine of prolyl endopeptidase. One labeled peptide was isolated and sequenced. The sequence surrounding the active-site serine was Asn-Gly-Gly-Ser-Asn-Gly-Gly. This sequence is different from the active-site sequences of other known serine proteases. This difference and the lack of overall homology with the known families of serine proteases suggest that prolyl endopeptidase represents a new type of serine protease.

  1. Single residue mutation in active site of serine acetyltransferase isoform 3 from Entamoeba histolytica assists in partial regaining of feedback inhibition by cysteine.

    Directory of Open Access Journals (Sweden)

    Sudhir Kumar

    Full Text Available The cysteine biosynthetic pathway is essential for survival of the protist pathogen Entamoeba histolytica, and functions by producing cysteine for countering oxidative attack during infection in human hosts. Serine acetyltransferase (SAT and O-acetylserine sulfhydrylase (OASS are involved in cysteine biosynthesis and are present in three isoforms each. While EhSAT1 and EhSAT2 are feedback inhibited by end product cysteine, EhSAT3 is nearly insensitive to such inhibition. The active site residues of EhSAT1 and of EhSAT3 are identical except for position 208, which is a histidine residue in EhSAT1 and a serine residue in EhSAT3. A combination of comparative modeling, multiple molecular dynamics simulations and free energy calculation studies showed a difference in binding energies of native EhSAT3 and of a S208H-EhSAT3 mutant for cysteine. Mutants have also been generated in vitro, replacing serine with histidine at position 208 in EhSAT3 and replacing histidine 208 with serine in EhSAT1. These mutants showed decreased affinity for substrate serine, as indicated by K(m, compared to the native enzymes. Inhibition kinetics in the presence of physiological concentrations of serine show that IC50 of EhSAT1 increases by about 18 folds from 9.59 µM for native to 169.88 µM for H208S-EhSAT1 mutant. Similar measurements with EhSAT3 confirm it to be insensitive to cysteine inhibition while its mutant (S208H-EhSAT3 shows a gain of cysteine inhibition by 36% and the IC50 of 3.5 mM. Histidine 208 appears to be one of the important residues that distinguish the serine substrate from the cysteine inhibitor.

  2. Roles of the active site residues and metal cofactors in noncanonical base-pairing during catalysis by human DNA polymerase iota.

    Science.gov (United States)

    Makarova, Alena V; Ignatov, Artem; Miropolskaya, Nataliya; Kulbachinskiy, Andrey

    2014-10-01

    Human DNA polymerase iota (Pol ι) is a Y-family polymerase that can bypass various DNA lesions but possesses very low fidelity of DNA synthesis in vitro. Structural analysis of Pol ι revealed a narrow active site that promotes noncanonical base-pairing during catalysis. To better understand the structure-function relationships in the active site of Pol ι we investigated substitutions of individual amino acid residues in its fingers domain that contact either the templating or the incoming nucleotide. Two of the substitutions, Y39A and Q59A, significantly decreased the catalytic activity but improved the fidelity of Pol ι. Surprisingly, in the presence of Mn(2+) ions, the wild-type and mutant Pol ι variants efficiently incorporated nucleotides opposite template purines containing modifications that disrupted either Hoogsteen or Watson-Crick base-pairing, suggesting that Pol ι may use various types of interactions during nucleotide addition. In contrast, in Mg(2+) reactions, wild-type Pol ι was dependent on Hoogsteen base-pairing, the Y39A mutant was essentially inactive, and the Q59A mutant promoted Watson-Crick interactions with template purines. The results suggest that Pol ι utilizes distinct mechanisms of nucleotide incorporation depending on the metal cofactor and reveal important roles of specific residues from the fingers domain in base-pairing and catalysis. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Residues at a Single Site Differentiate Animal Cryptochromes from Cyclobutane Pyrimidine Dimer Photolyases by Affecting the Proteins' Preferences for Reduced FAD.

    Science.gov (United States)

    Xu, Lei; Wen, Bin; Wang, Yuan; Tian, Changqing; Wu, Mingcai; Zhu, Guoping

    2017-06-19

    Cryptochromes (CRYs) and photolyases belong to the cryptochrome/photolyase family (CPF). Reduced FAD is essential for photolyases to photorepair UV-induced cyclobutane pyrimidine dimers (CPDs) or 6-4 photoproducts in DNA. In Drosophila CRY (dCRY, a type I animal CRY), FAD is converted to the anionic radical but not to the reduced state upon illumination, which might induce a conformational change in the protein to relay the light signal downstream. To explore the foundation of these differences, multiple sequence alignment of 650 CPF protein sequences was performed. We identified a site facing FAD (Ala377 in Escherichia coli CPD photolyase and Val415 in dCRY), hereafter referred to as "site 377", that was distinctly conserved across these sequences: CPD photolyases often had Ala, Ser, or Asn at this site, whereas animal CRYs had Ile, Leu, or Val. The binding affinity for reduced FAD, but not the photorepair activity of E. coli photolyase, was dramatically impaired when replacing Ala377 with any of the three CRY residues. Conversely, in V415S and V415N mutants of dCRY, FAD was photoreduced to its fully reduced state after prolonged illumination, and light-dependent conformational changes of these mutants were severely inhibited. We speculate that the residues at site 377 play a key role in the different preferences of CPF proteins for reduced FAD, which differentiate animal CRYs from CPD photolyases. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Photoactivation of the BLUF protein PixD Probed by the Site-Specific Incorporation of Fluorotyrosine Residues

    KAUST Repository

    Gil, Agnieszka A.

    2017-09-06

    The flavin chromophore in blue light using FAD (BLUF) photoreceptors is surrounded by a hydrogen bond network that senses and responds to changes in the electronic structure of the flavin on the ultrafast time scale. The hydrogen bond network includes a strictly conserved Tyr residue, and previously we explored the role of this residue, Y21, in the photoactivation mechanism of the BLUF protein AppA by the introduction of fluorotyrosine (F-Tyr) analogs that modulated the pKa and reduction potential of Y21 by 3.5 pH units and 200 mV, respectively. Although little impact on the forward (dark to light adapted form) photoreaction was observed, the change in Y21 pKa led to a 4,000-fold increase in the rate of dark state recovery. In the present work we have extended these studies to the BLUF protein PixD, where, in contrast to AppA, modulation in the Tyr (Y8) pKa has a profound impact on the forward photoreaction. In particular, a decrease in Y8 pKa by 2 or more pH units prevents formation of a stable light state, consistent with a photoactivation mechanism that involves proton transfer or proton coupled electron transfer from Y8 to the electronically excited FAD. Conversely, the effect of pKa on the rate of dark recovery is markedly reduced in PixD. These observations highlight very significant differences between the photocycles of PixD and AppA, despite their sharing highly conserved FAD binding architectures.

  5. Photoactivation of the BLUF protein PixD Probed by the Site-Specific Incorporation of Fluorotyrosine Residues

    KAUST Repository

    Gil, Agnieszka A.; Laptenok, Sergey P.; Iuliano, James N.; Lukacs, Andras; Verma, Anil; Hall, Christopher R.; Yoon, EunBin; Brust, Richard; Greetham, Gregory M.; Towrie, Michael; French, Jarrod B.; Meech, Stephen R.; Tonge, Peter J

    2017-01-01

    The flavin chromophore in blue light using FAD (BLUF) photoreceptors is surrounded by a hydrogen bond network that senses and responds to changes in the electronic structure of the flavin on the ultrafast time scale. The hydrogen bond network includes a strictly conserved Tyr residue, and previously we explored the role of this residue, Y21, in the photoactivation mechanism of the BLUF protein AppA by the introduction of fluorotyrosine (F-Tyr) analogs that modulated the pKa and reduction potential of Y21 by 3.5 pH units and 200 mV, respectively. Although little impact on the forward (dark to light adapted form) photoreaction was observed, the change in Y21 pKa led to a 4,000-fold increase in the rate of dark state recovery. In the present work we have extended these studies to the BLUF protein PixD, where, in contrast to AppA, modulation in the Tyr (Y8) pKa has a profound impact on the forward photoreaction. In particular, a decrease in Y8 pKa by 2 or more pH units prevents formation of a stable light state, consistent with a photoactivation mechanism that involves proton transfer or proton coupled electron transfer from Y8 to the electronically excited FAD. Conversely, the effect of pKa on the rate of dark recovery is markedly reduced in PixD. These observations highlight very significant differences between the photocycles of PixD and AppA, despite their sharing highly conserved FAD binding architectures.

  6. Triazacyclophane (TAC)-scaffolded histidine and aspartic acid residues as mimics of non-heme metalloenzyme active sites

    NARCIS (Netherlands)

    Albada, H.B.; Soulimani, F.; Jacobs, H.J.F.; Versluis, C.; Weckhuysen, B.M.; Liskamp, R.M.J.

    2012-01-01

    We describe the synthesis and coordination behaviour to copper(II) of two close structural triazacyclophane-based mimics of two often encountered aspartic acid and histidine containing metalloenzyme active sites. Coordination of these mimics to copper(I) and their reaction with molecular oxygen

  7. An evaluation of 1957 to 1966 data to assess residual radiation from nuclear weapons sites in the U.S.

    OpenAIRE

    Cheng, Chih Jen

    2016-01-01

    Between 1950s and 1960s, active bomb tests produced many radionuclides and exposed radiation to the environment. This thesis addresses the radioactivity caused by strontium-90 (90Sr), a beta-only emitter and the health concerns it has caused. Through the bomb detonations, radioactive compounds were carried by atmospheric transport globally. This research is conducted to analyze the pattern of distribution and the inventory of 90Sr. Data from a total of 53 sites were collected from public reco...

  8. Biodegradation of Aged Residues of Atrazine and Alachlor in a Mix-Load Site Soil by Fungal Enzymes

    Directory of Open Access Journals (Sweden)

    Anastasia E. M. Chirnside

    2011-01-01

    Full Text Available Soils from bulk pesticide mixing and loading (mix-load sites are often contaminated with a complex mixture of pesticides, herbicides, and other organic compounds used in pesticide formulations that limits the success of remediation efforts. Therefore, there is a need to find remediation strategies that can successfully clean up these mix-load site soils. This paper examined the degradation of atrazine (2-chloro-4-ethylamino-6-isopropylamino-S-triazine; AT and alachlor (2-chloro-2, 6-diethyl-N-[methoxymethyl]-acetanilide in contaminated mix-load site soil utilizing an extracellular fungal enzyme solution derived from the white rot fungus, Phanerochaete chrysosporium, grown in a packed bed bioreactor. Thirty-two percent of AT and 54% of AL were transformed in the biometers. The pseudo first-order rate constant for AT and AL biodegradation was 0.0882 d−1 and 0.2504 d−1, respectively. The half-life (1/2 for AT and AL was 8.0 and 3.0 days, respectively. Compared to AT, the initial disappearance of AL proceeded at a faster rate and resulted in a greater amount of AL transformed. Based on the net Co2 evolved from the biometers, about 4% of the AT and AL initially present in the soil was completely mineralized.

  9. Localization of α1-2 Fucose Glycan in the Mouse Olfactory Pathway.

    Science.gov (United States)

    Kondoh, Daisuke; Kamikawa, Akihiro; Sasaki, Motoki; Kitamura, Nobuo

    2017-01-01

    Glycoconjugates in the olfactory system play critical roles in neuronal formation, and α1-2 fucose (α1-2Fuc) glycan mediates neurite outgrowth and synaptic plasticity. Histochemical findings of α1-2Fuc glycan in the mouse olfactory system detected using Ulex europaeus agglutinin-I (UEA-I) vary. This study histochemically assessed the main olfactory and vomeronasal pathways in male and female ICR and C57BL/6J mice aged 3-4 months using UEA-I. Ulex europaeus agglutinin-I reacted with most receptor cells arranged mainly at the basal region of the olfactory epithelium. The olfactory nerve layer and glomerular layer of the main olfactory bulb were speckled with positive UEA-I staining, and positive fibers were scattered from the glomerular to the internal plexiform layer. The lateral olfactory tract and rostral migratory stream were also positive for UEA-I. We identified superficial short-axon cells, interneurons of the external plexiform layer, external, middle and internal tufted cells, mitral cells and granule cells as the origins of the UEA-I-positive fibers in the main olfactory bulb. The anterior olfactory nucleus, anterior piriform cortex and olfactory tubercle were negative for UEA-I. Most receptor cells in the vomeronasal epithelium and most glomeruli of the accessory olfactory bulb were positive for UEA-I. Our findings indicated that α1-2Fuc glycan is located within the primary and secondary, but not the ternary, pathways of the main olfactory system, in local circuits of the main olfactory bulb and within the primary, but not secondary, pathway of the vomeronasal system. © 2016 S. Karger AG, Basel.

  10. Mechanism of Flavoprotein l-6-Hydroxynicotine Oxidase: pH and Solvent Isotope Effects and Identification of Key Active Site Residues.

    Science.gov (United States)

    Fitzpatrick, Paul F; Chadegani, Fatemeh; Zhang, Shengnan; Dougherty, Vi

    2017-02-14

    The flavoenzyme l-6-hydroxynicotine oxidase is a member of the monoamine oxidase family that catalyzes the oxidation of (S)-6-hydroxynicotine to 6-hydroxypseudooxynicotine during microbial catabolism of nicotine. While the enzyme has long been understood to catalyze oxidation of the carbon-carbon bond, it has recently been shown to catalyze oxidation of a carbon-nitrogen bond [Fitzpatrick, P. F., et al. (2016) Biochemistry 55, 697-703]. The effects of pH and mutagenesis of active site residues have now been utilized to study the mechanism and roles of active site residues. Asn166 and Tyr311 bind the substrate, while Lys287 forms a water-mediated hydrogen bond with flavin N5. The N166A and Y311F mutations result in ∼30- and ∼4-fold decreases in k cat /K m and k red for (S)-6-hydroxynicotine, respectively, with larger effects on the k cat /K m value for (S)-6-hydroxynornicotine. The K287M mutation results in ∼10-fold decreases in these parameters and a 6000-fold decrease in the k cat /K m value for oxygen. The shapes of the pH profiles are not altered by the N166A and Y311F mutations. There is no solvent isotope effect on the k cat /K m value for amines. The results are consistent with a model in which both the charged and neutral forms of the amine can bind, with the former rapidly losing a proton to a hydrogen bond network of water and amino acids in the active site prior to the transfer of hydride to the flavin.

  11. Role of allosteric switch residue histidine 195 in maintaining active-site asymmetry in presynaptic filaments of bacteriophage T4 UvsX recombinase.

    Science.gov (United States)

    Farb, Joshua N; Morrical, Scott W

    2009-01-16

    Recombinases of the highly conserved RecA/Rad51 family play central roles in homologous recombination and DNA double-stranded break repair. RecA/Rad51 enzymes form presynaptic filaments on single-stranded DNA (ssDNA) that are allosterically activated to catalyze ATPase and DNA strand-exchange reactions. Information is conveyed between DNA- and ATP-binding sites, in part, by a highly conserved glutamine residue (Gln194 in Escherichia coli RecA) that acts as an allosteric switch. The T4 UvsX protein is a divergent RecA ortholog and contains histidine (His195) in place of glutamine at the allosteric switch position. UvsX and RecA catalyze similar strand-exchange reactions, but differ in other properties. UvsX produces both ADP and AMP as products of its ssDNA-dependent ATPase activity--a property that is unique among characterized recombinases. Details of the kinetics of ssDNA-dependent ATP hydrolysis reactions indicate that UvsX-ssDNA presynaptic filaments are asymmetric and contain two classes of ATPase active sites: one that generates ADP, and another that generates AMP. Active-site asymmetry is reduced by mutations at the His195 position, since UvsX-H195Q and UvsX-H195A mutants both exhibit stronger ssDNA-dependent ATPase activity, with lower cooperativity and markedly higher ADP/AMP product ratios, than wild-type UvsX. Reduced active-site asymmetry correlates strongly with reduced ssDNA-binding affinity and DNA strand-exchange activity in both H195Q and H195A mutants. These and other results support a model in which allosteric switch residue His195 controls the formation of an asymmetric conformation of UvsX-ssDNA filaments that is active in DNA strand exchange. The implications of our findings for UvsX recombination functions, and for RecA functions in general, are discussed.

  12. The dapE-encoded N-succinyl-L,L-Diaminopimelic Acid Desuccinylase from Haemophilus influenzae Contains two Active Site Histidine Residues

    Science.gov (United States)

    Gillner, Danuta M.; Bienvenue, David L.; Nocek, Boguslaw P.; Joachimiak, Andrzej; Zachary, Vincentos; Bennett, Brian; Holz, Richard C.

    2009-01-01

    The catalytic and structural properties of the H67A and H349A altered dapE-encoded N-succinyl-l,l-diaminopimelic acid desuccinylase (DapE) from H. influenzae were investigated. Based on sequence alignment with CPG2 both H67 and H349 were predicted to be Zn(II) ligands. Catalytic activity was observed for the H67A altered DapE enzyme which exhibited kcat = 1.5 ± 0.5 sec−1 and Km = 1.4 ± 0.3 mM. No catalytic activity was observed for H349A under the experimental conditions used. The EPR and electronic absorption data indicate that the Co(II) ion bound to H349A-DapE is analogous to WT DapE after the addition of a single Co(II) ion. The addition of one equivalent of Co(II) to H67A altered DapE provides spectra that are very different from the first Co(II) binding site of the WT enzyme, but similar to the second binding site. The EPR and electronic absorption data, in conjunction with the kinetic data, are consistent with the assignment of H67 and H349 as active site metal ligands for the DapE from H. influenzae. Furthermore, the data suggest that H67 is a ligand in the first metal binding site while H349 resides in the second metal binding site. A three-dimensional homology structure of the DapE from H. influenzae was generated using the X-ray crystal structure of the DapE from N. meningitidis as a template and superimposed on the structure of AAP. This homology structure confirms the assignment of H67 and H349 as active site ligands. The superimposition of the homology model of DapE with the dizinc(II) structure of AAP indicates that within 4.0 Å of the Zn(II) binding sites of AAP, all of the amino acid residues of DapE are nearly identical. PMID:18712420

  13. The dapE-encoded N-succinyl-L,L-diaminopimelic acid desuccinylase from Haemophilus influenzae contains two active-site histidine residues.

    Science.gov (United States)

    Gillner, Danuta M; Bienvenue, David L; Nocek, Boguslaw P; Joachimiak, Andrzej; Zachary, Vincentos; Bennett, Brian; Holz, Richard C

    2009-01-01

    The catalytic and structural properties of the H67A and H349A dapE-encoded N-succinyl-L,L-diaminopimelic acid desuccinylase (DapE) from Haemophilus influenzae were investigated. On the basis of sequence alignment with the carboxypeptidase from Pseudomonas sp. strain RS-16, both H67 and H349 were predicted to be Zn(II) ligands. The H67A DapE enzyme exhibited a decreased catalytic efficiency (180-fold) compared with wild-type (WT) DapE towards N-succinyldiaminopimelic acid. No catalytic activity was observed for H349A under the experimental conditions used. The electronic paramagnetic resonance (EPR) and electronic absorption data indicate that the Co(II) ion bound to H349A-DapE is analogous to that of WT DapE after the addition of a single Co(II) ion. The addition of 1 equiv of Co(II) to H67A DapE provides spectra that are very different from those of the first Co(II) binding site of the WT enzyme, but that are similar to those of the second binding site. The EPR and electronic absorption data, in conjunction with the kinetic data, are consistent with the assignment of H67 and H349 as active-site metal ligands for the DapE from H. influenzae. Furthermore, the data suggest that H67 is a ligand in the first metal binding site, while H349 resides in the second metal binding site. A three-dimensional homology structure of the DapE from H. influenzae was generated using the X-ray crystal structure of the DapE from Neisseria meningitidis as a template and superimposed on the structure of the aminopeptidase from Aeromonas proteolytica (AAP). This homology structure confirms the assignment of H67 and H349 as active-site ligands. The superimposition of the homology model of DapE with the dizinc(II) structure of AAP indicates that within 4.0 A of the Zn(II) binding sites of AAP all of the amino acid residues of DapE are nearly identical.

  14. Conversion of cheese whey into a fucose- and glucuronic acid-rich extracellular polysaccharide by Enterobacter A47.

    Science.gov (United States)

    Antunes, Sílvia; Freitas, Filomena; Alves, Vítor D; Grandfils, Christian; Reis, Maria A M

    2015-09-20

    Cheese whey was used as the sole substrate for the production of extracellular polysaccharides (EPS) by Enterobacter A47. An EPS concentration of 6.40 g L(-1) was reached within 3.2 days of cultivation, corresponding to a volumetric productivity of 2.00 g L(-1) d(-1). The produced EPS was mainly composed of glucuronic acid (29 mol%) and fucose (29 mol%), with lower contents of glucose and galactose (21 mol% each) and a total acyl groups content of 32 wt.%. The polymer had an average molecular weight of 1.8×10(6) Da, with a polydispersity index of 1.2, and an intrinsic viscosity of 8.0 dL g(-1). EPS aqueous solutions (1.0 wt.% in 0.01 M NaCl, at pH 8.0) presented a shear thinning behavior with a viscosity of the first Newtonian plateau approaching 0.1 Pas. This novel glucuronic acid-rich polymer possesses interesting rheological properties, which, together with its high content of glucuronic acid and fucose, two bioactive sugar monomers, confers it a great potential for use in high-value applications, such as cosmetics and pharmaceuticals. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Effects of mannose, fructose, and fucose on the structure, stability, and hydration of lysozyme in aqueous solution

    DEFF Research Database (Denmark)

    Rahim, Abdoul; Peters, Günther H.J.; Jalkanen, Karl J.

    2013-01-01

    The bio-protective properties of monosaccharaides, namely mannose, fructose and fucose, on the stability and dynamical properties of the NMR determined hen egg-white lysozyme structure have been investigated by means of molecular dynamics simulations at room temperature in aqueous solution and in...... of the solvent and sugar distributions around lysozyme was used to investigate the interfacial solvent and sugar structure near the protein surface.......The bio-protective properties of monosaccharaides, namely mannose, fructose and fucose, on the stability and dynamical properties of the NMR determined hen egg-white lysozyme structure have been investigated by means of molecular dynamics simulations at room temperature in aqueous solution and in 7...... and 13 wt % concentrations of the three sugars. Results are discussed in the framework of the bio-protective phenomena. The three sugars show similar bio-protective behaviours at room temperature (300 K) in the concentration range studied as shown by the small RMSDs of the resulting MD structures from...

  16. A method for predicting individual residue contributions to enzyme specificity and binding-site energies, and its application to MTH1.

    Science.gov (United States)

    Stewart, James J P

    2016-11-01

    A new method for predicting the energy contributions to substrate binding and to specificity has been developed. Conventional global optimization methods do not permit the subtle effects responsible for these properties to be modeled with sufficient precision to allow confidence to be placed in the results, but by making simple alterations to the model, the precisions of the various energies involved can be improved from about ±2 kcal mol -1 to ±0.1 kcal mol -1 . This technique was applied to the oxidized nucleotide pyrophosphohydrolase enzyme MTH1. MTH1 is unusual in that the binding and reaction sites are well separated-an advantage from a computational chemistry perspective, as it allows the energetics involved in docking to be modeled without the need to consider any issues relating to reaction mechanisms. In this study, two types of energy terms were investigated: the noncovalent interactions between the binding site and the substrate, and those responsible for discriminating between the oxidized nucleotide 8-oxo-dGTP and the normal dGTP. Both of these were investigated using the semiempirical method PM7 in the program MOPAC. The contributions of the individual residues to both the binding energy and the specificity of MTH1 were calculated by simulating the effect of mutations. Where comparisons were possible, all calculated results were in agreement with experimental observations. This technique provides fresh insight into the binding mechanism that enzymes use for discriminating between possible substrates.

  17. Evolutionary conservativeness of electric field in the Cu,Zn superoxide dismutase active site. Evidence for co-ordinated mutation of charged amino acid residues.

    Science.gov (United States)

    Desideri, A; Falconi, M; Polticelli, F; Bolognesi, M; Djinovic, K; Rotilio, G

    1992-01-05

    Equipotential lines were calculated, using the Poisson-Boltzmann equation, for six Cu,Zn superoxide dismutases with different protein electric charge and various degrees of sequence homology, namely those from ox, pig, sheep, yeast, and the isoenzymes A and B from the amphibian Xenopus laevis. The three-dimensional structures of the porcine and ovine superoxide dismutases were obtained by molecular modelling reconstruction using the structure of the highly homologous bovine enzyme as a template. The three-dimensional structure of the evolutionary distant yeast Cu,Zn superoxide dismutase was recently resolved by us, while computer-modelled structures are available for X. laevis isoenzymes. The six proteins display large differences in the net protein charge and distribution of electrically charged surface residues but the trend of the equipotential lines in the proximity of the active sites was found to be constant in all cases. These results are in line with the very similar catlytic rate constants experimentally measured for the corresponding enzyme activities. This analysis shows that electrostatic guidance for the enzyme-substrate interaction in Cu,Zn superoxide dismutases is related to a spatial distribution of charges, arranged so as to maintain, in the area surrounding the active sites, an identical electrostatic potential distribution, which is conserved in the evolution of this protein family.

  18. Using residual stacking to mitigate site-specific errors in order to improve the quality of GNSS-based coordinate time series of CORS

    Science.gov (United States)

    Knöpfler, Andreas; Mayer, Michael; Heck, Bernhard

    2014-05-01

    Within the last decades, positioning using GNSS (Global Navigation Satellite Systems; e.g., GPS) has become a standard tool in many (geo-) sciences. The positioning methods Precise Point Positioning and differential point positioning based on carrier phase observations have been developed for a broad variety of applications with different demands for example on accuracy. In high precision applications, a lot of effort was invested to mitigate different error sources: the products for satellite orbits and satellite clocks were improved; the misbehaviour of satellite and receiver antennas compared to an ideal antenna is modelled by calibration values on absolute level, the modelling of the ionosphere and the troposphere is updated year by year. Therefore, within processing of data of CORS (continuously operating reference sites), equipped with geodetic hardware using a sophisticated strategy, the latest products and models nowadays enable positioning accuracies at low mm level. Despite the considerable improvements that have been achieved within GNSS data processing, a generally valid multipath model is still lacking. Therefore, site specific multipath still represents a major error source in precise GNSS positioning. Furthermore, the calibration information of receiving GNSS antennas, which is for instance derived by a robot or chamber calibration, is valid strictly speaking only for the location of the calibration. The calibrated antenna can show a slightly different behaviour at the CORS due to near field multipath effects. One very promising strategy to mitigate multipath effects as well as imperfectly calibrated receiver antennas is to stack observation residuals of several days, thereby, multipath-loaded observation residuals are analysed for example with respect to signal direction, to find and reduce systematic constituents. This presentation will give a short overview about existing stacking approaches. In addition, first results of the stacking approach

  19. Incorporation of carbohydrate residues into peroxidase isoenzymes in horseradish roots.

    Science.gov (United States)

    Lew, J Y; Shannon, L M

    1973-11-01

    Sliced root tissue of the horseradish plant (Armoracia rusticana), when incubated with mannose-U-(14)C, incorporated radioactivity into peroxidase isoenzymes. Over 90% of the radioactivity in the highly purified peroxidase isoenzymes was present in the neutral sugar residues of the molecule, i.e. fucose, arabinose, xylose, mannose. When the root slices were incubated simultaneously with leucine-4,5-(3)H and mannose-U-(14)C, cycloheximide strongly inhibited leucine incorporation into the peptide portion of peroxidase isoenzymes but had little effect on the incorporation of (14)C into the neutral sugars. These results indicated that synthesis of the peptide portion of peroxidase was completed before the monosaccharide residues were attached to the molecule. This temporal relationship between the synthesis of protein and the attachment of carbohydrate residues in the plant glycoprotein, horseradish peroxidase, appears to be similar to that reported for glycoprotein biosynthesis in many mammalian systems.

  20. Construction of a catalytically inactive cholesterol oxidase mutant: investigation of the interplay between active site-residues glutamate 361 and histidine 447.

    Science.gov (United States)

    Yin, Ye; Liu, Pingsheng; Anderson, Richard G W; Sampson, Nicole S

    2002-06-15

    Cholesterol oxidase catalyzes the oxidation of cholesterol to cholest-5-en-3-one and its subsequent isomerization into cholest-4-en-3-one. Two active-site residues, His447 and Glu361, are important for catalyzing the oxidation and isomerization reactions, respectively. Double-mutants were constructed to test the interplay between these residues in catalysis. We observed that the k(cat) of oxidation for the H447Q/E361Q mutant was 3-fold less than that for H447Q and that the k(cat) of oxidation for the H447E/E361Q mutant was 10-fold slower than that for H447E. Because both doubles-mutants do not have a carboxylate at position 361, they do not catalyze isomerization of the reaction intermediate cholest-5-en-3-one to cholest-4-en-3-one. These results suggest that Glu361 can compensate for the loss of histidine at position 447 by acting as a general base catalyst for oxidation of cholesterol. Importantly, the construction of the double-mutant H447E/E361Q yields an enzyme that is 31,000-fold slower than wild type in k(cat) for oxidation. The H447E/E361Q mutant is folded like native enzyme and still associates with model membranes. Thus, this mutant may be used to study the effects of membrane binding in the absence of catalytic activity. It is demonstrated that in assays with caveolae membrane fractions, the wild-type enzyme uncouples platelet-derived growth factor receptor beta (PDGFRbeta) autophosphorylation from tyrosine phosphorylation of neighboring proteins, and the H447E/E361Q mutant does not. Thus maintenance of membrane structure by cholesterol is important for PDGFRbeta-mediated signaling. The cholesterol oxidase mutant probe described will be generally useful for investigating the role of membrane structure in signal transduction pathways in addition to the PDGFRbeta-dependent pathway tested.

  1. HEU Measurements of Holdup and Recovered Residue in the Deactivation and Decommissioning Activities of the 321-M Reactor Fuel Fabrication Facility at the Savannah River Site

    Energy Technology Data Exchange (ETDEWEB)

    DEWBERRY, RAYMOND; SALAYMEH, SALEEM R.; CASELLA, VITO R.; MOORE, FRANK S.

    2005-03-11

    This paper contains a summary of the holdup and material control and accountability (MC&A) assays conducted for the determination of highly enriched uranium (HEU) in the deactivation and decommissioning (D&D) of Building 321-M at the Savannah River Site (SRS). The 321-M facility was the Reactor Fuel Fabrication Facility at SRS and was used to fabricate HEU fuel assemblies, lithium-aluminum target tubes, neptunium assemblies, and miscellaneous components for the SRS production reactors. The facility operated for more than 35 years. During this time thousands of uranium-aluminum-alloy (U-Al) production reactor fuel tubes were produced. After the facility ceased operations in 1995, all of the easily accessible U-Al was removed from the building, and only residual amounts remained. The bulk of this residue was located in the equipment that generated and handled small U-Al particles and in the exhaust systems for this equipment (e.g., Chip compactor, casting furnaces, log saw, lathes A & B, cyclone separator, Freon{trademark} cart, riser crusher, ...etc). The D&D project is likely to represent an important example for D&D activities across SRS and across the Department of Energy weapons complex. The Savannah River National Laboratory was tasked to conduct holdup assays to quantify the amount of HEU on all components removed from the facility prior to placing in solid waste containers. The U-235 holdup in any single component of process equipment must not exceed 50 g in order to meet the container limit. This limit was imposed to meet criticality requirements of the low level solid waste storage vaults. Thus the holdup measurements were used as guidance to determine if further decontamination of equipment was needed to ensure that the quantity of U-235 did not exceed the 50 g limit and to ensure that the waste met the Waste Acceptance Criteria (WAC) of the solid waste storage vaults. Since HEU is an accountable nuclear material, the holdup assays and assays of recovered

  2. Natural vitamin B12 and fucose supplementation of green smoothies with edible algae and related quality changes during their shelf life.

    Science.gov (United States)

    Castillejo, Noelia; Martínez-Hernández, Ginés Benito; Goffi, Valentina; Gómez, Perla A; Aguayo, Encarna; Artés, Francisco; Artés-Hernández, Francisco

    2018-04-01

    Some algae are an excellent sources of vitamin B12, of special interest for vegetarian/vegan consumers, and of fucose to supplement fruit and vegetable beverages such as smoothies. Nevertheless, supplementation of smoothies with algae may lead to possible quality changes during smoothie shelf life that need to be studied. Therefore, the quality changes in fresh green smoothies supplemented (2.2%) with nine edible algae (sea lettuce, kombu, wakame, thongweed, dulse, Irish moss, nori, Spirulina and Chlorella) were studied throughout 24 days at 5 °C. The initial vitamin C content - 238.7-326.0 mg kg -1 fresh weight (FW) - of a 200 g portion of any of the smoothies ensured full coverage of its recommended daily intake, and still supplying 50-60% of the recommended intake after 7 days. Chlorella and Spirulina smoothies showed the highest vitamin B12 content (33.3 and 15.3 µg kg -1 FW, respectively), while brown algae showed fucose content of 141.1-571.3 mg kg -1 FW. These vitamin B12 and fucose contents were highly maintained during shelf life. The Spirulina supplementation of a 200 g smoothie portion ensured full coverage of the recommended vitamin B12 intake, with lower vitamin C degradation, during a shelf life of 17 days. Furthermore, thongweed and kombu are also considered as excellent fucose sources with similar shelf life. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  3. Autoradiographic demonstration of 3H-labelled glycoproteins in 'light' and 'dark' neurons of different grisea of rat brain after intraventricular application of tritiated fucose

    International Nuclear Information System (INIS)

    Froehlich, J.; Rummelfaenger, H.; Pohle, W.

    1982-01-01

    The simultaneous demonstration of 3 H radioactivity and of 'light' and 'dark' neurons by histological staining revealed that the occurrence of 'light' and 'dark' neurons are morphological correlates of different activity stages of cell metabolism. In this connection it became evident that 'light' nerve cells incorporated significantly greater amounts of fucose and therefore should be regarded as metabolically more active. (author)

  4. Site-directed fluorescence labeling reveals a revised N-terminal membrane topology and functional periplasmic residues in the Escherichia coli cell division protein FtsK.

    Science.gov (United States)

    Berezuk, Alison M; Goodyear, Mara; Khursigara, Cezar M

    2014-08-22

    In Escherichia coli, FtsK is a large integral membrane protein that coordinates chromosome segregation and cell division. The N-terminal domain of FtsK (FtsKN) is essential for division, and the C terminus (FtsKC) is a well characterized DNA translocase. Although the function of FtsKN is unknown, it is suggested that FtsK acts as a checkpoint to ensure DNA is properly segregated before septation. This may occur through modulation of protein interactions between FtsKN and other division proteins in both the periplasm and cytoplasm; thus, a clear understanding of how FtsKN is positioned in the membrane is required to characterize these interactions. The membrane topology of FtsKN was initially determined using site-directed reporter fusions; however, questions regarding this topology persist. Here, we report a revised membrane topology generated by site-directed fluorescence labeling. The revised topology confirms the presence of four transmembrane segments and reveals a newly identified periplasmic loop between the third and fourth transmembrane domains. Within this loop, four residues were identified that, when mutated, resulted in the appearance of cellular voids. High resolution transmission electron microscopy of these voids showed asymmetric division of the cytoplasm in the absence of outer membrane invagination or visible cell wall ingrowth. This uncoupling reveals a novel role for FtsK in linking cell envelope septation events and yields further evidence for FtsK as a critical checkpoint of cell division. The revised topology of FtsKN also provides an important platform for future studies on essential interactions required for this process. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  5. Single molecule TPM analysis of the catalytic pentad mutants of Cre and Flp site-specific recombinases: contributions of the pentad residues to the pre-chemical steps of recombination

    Science.gov (United States)

    Fan, Hsiu-Fang; Cheng, Yong-Song; Ma, Chien-Hui; Jayaram, Makkuni

    2015-01-01

    Cre and Flp site-specific recombinase variants harboring point mutations at their conserved catalytic pentad positions were characterized using single molecule tethered particle motion (TPM) analysis. The findings reveal contributions of these amino acids to the pre-chemical steps of recombination. They suggest functional differences between positionally conserved residues in how they influence recombinase-target site association and formation of ‘non-productive’, ‘pre-synaptic’ and ‘synaptic’ complexes. The most striking difference between the two systems is noted for the single conserved lysine. The pentad residues in Cre enhance commitment to recombination by kinetically favoring the formation of pre-synaptic complexes. These residues in Flp serve a similar function by promoting Flp binding to target sites, reducing non-productive binding and/or enhancing the rate of assembly of synaptic complexes. Kinetic comparisons between Cre and Flp, and between their derivatives lacking the tyrosine nucleophile, are consistent with a stronger commitment to recombination in the Flp system. The effect of target site orientation (head-to-head or head-to-tail) on the TPM behavior of synapsed DNA molecules supports the selection of anti-parallel target site alignment prior to the chemical steps. The integrity of the synapse, whose establishment/stability is fostered by strand cleavage in the case of Flp but not Cre, appears to be compromised by the pentad mutations. PMID:25765648

  6. Effects of mutagenesis of aspartic acid residues in the putative phosphoribosyl diphosphate binding site of Escherichia coli phosphoribosyl diphosphate synthetase on metal ion specificity and ribose-5-phosphate binding

    DEFF Research Database (Denmark)

    Willemoës, Martin; Nilsson, Dan; Hove-Jensen, Bjarne

    1996-01-01

    The three conserved aspartic acid residues of the 5-phospho-d-ribosyl a-1-diphosphate binding site (213-GRDCVLVDDMIDTGGT-228) of Escherichia coli phosphoribosyl diphosphate synthetase were studied by analysis of the mutant enzymes D220E, D220F, D221A, D224A, and D224S. The mutant enzymes showed...... enzymes were dependent on the metal ion present, suggesting a function of the investigated aspartic acid residues both in the binding of ribose 5-phosphate, possibly via a divalent metal ion, and in the interaction with a divalent metal ion during catalysis....

  7. GDP-L-fucose: .beta.-D-galactoside 2-.alpha.-L-fucosyltransferases, DNA sequences encoding the same, method for producing the same and a method of genotyping a person

    Science.gov (United States)

    Lowe, John B.; Lennon, Gregory; Rouquier, Sylvie; Giorgi, Dominique; Kelly, Robert J.

    1998-01-01

    The gene encoding GDP-L-fucose: .beta.-D-Galactoside 2-.alpha.-L-fucosyltransferase has been cloned, and a mutation in this gene has been found to be responsible for an individual being a non-secretor.

  8. GDP-L-fucose: {beta}-D-galactoside 2-{alpha}-Lfucosyltransferases, DNA sequences encoding the same, method for producing the same and a method of genotyping a person

    Science.gov (United States)

    Lowe, J.B.; Lennon, G.; Rouquier, S.; Giorgi, D.; Kelly, R.J.

    1998-09-15

    The gene encoding GDP-L-fucose: {beta}-D-Galactoside 2-{alpha}-Lfucosyltransferase has been cloned, and a mutation in this gene has been found to be responsible for an individual being a non-secretor. 30 figs.

  9. Measures for radiation prevention and remediation of islightly radioactive contaminated sites by phytoremediation and subsequent utilization of the loaded plant residues (PHYTOREST). Final report

    International Nuclear Information System (INIS)

    Willscher, Sabine; Werner, Peter; Jablonski, Lukasz; Wittig, Juliane

    2013-01-01

    contaminated geosubstrates now can be carried out within the radiation protection regulations. Hence, the project provides a substantial contribution to the radiation protection of HM/R contaminated soils. Within the research project, ways for the utilization of HM/R- contaminated plant residuals were highlighted; this gives a substantial contribution for minimization of wastes, the winning of sustainable bioenergy and the recycling of materials. Here, different ways of solutions were investigated. The research project was carried out within the scientific funding program ''Closedown and decommissioning of nuclear facilities''. The results of the project will contribute to the development of a biologically benign, sustainable technique for the remediation of large contaminated areas that originate mostly from the legacy of the former U mining. As a general result of this comprehensive research project, a phytostabilization/ phytoextraction of such SM/R contaminated sites is feasible with a protection of ground water, and the plant crop from phytoremediation of the HM/R contaminated field site can be utilized for the winning of bioenergy (gaseous/ liquid products or thermal utilization). The beneficial combination of phytoremediation and subsequent utilization of the biomass can be further developed to an innovative and sustainable remediation technology with national and international application potential.

  10. Characterization Report on Sand, Slag, and Crucible Residues and on Fluoride Residues

    International Nuclear Information System (INIS)

    Murray, A.M.

    1999-01-01

    This paper reports on the chemical characterization of the sand, slag, and crucible (SS and C) residues and the fluoride residues that may be shipped from the Rocky Flats Environmental Technology Site (RFETS) to Savannah River Site (SRS)

  11. Oxidized amino acid residues in the vicinity of Q(A and Pheo(D1 of the photosystem II reaction center: putative generation sites of reducing-side reactive oxygen species.

    Directory of Open Access Journals (Sweden)

    Laurie K Frankel

    Full Text Available Under a variety of stress conditions, Photosystem II produces reactive oxygen species on both the reducing and oxidizing sides of the photosystem. A number of different sites including the Mn4O5Ca cluster, P680, PheoD1, QA, QB and cytochrome b559 have been hypothesized to produce reactive oxygen species in the photosystem. In this communication using Fourier-transform ion cyclotron resonance mass spectrometry we have identified several residues on the D1 and D2 proteins from spinach which are oxidatively modified and in close proximity to QA (D1 residues (239F, (241Q, (242E and the D2 residues (238P, (239T, (242E and (247M and PheoD1 (D1 residues (130E, (133L and (135F. These residues may be associated with reactive oxygen species exit pathways located on the reducing side of the photosystem, and their modification may indicate that both QA and PheoD1 are sources of reactive oxygen species on the reducing side of Photosystem II.

  12. O-Fucose Monosaccharide of Drosophila Notch Has a Temperature-sensitive Function and Cooperates with O-Glucose Glycan in Notch Transport and Notch Signaling Activation*

    Science.gov (United States)

    Ishio, Akira; Sasamura, Takeshi; Ayukawa, Tomonori; Kuroda, Junpei; Ishikawa, Hiroyuki O.; Aoyama, Naoki; Matsumoto, Kenjiroo; Gushiken, Takuma; Okajima, Tetsuya; Yamakawa, Tomoko; Matsuno, Kenji

    2015-01-01

    Notch (N) is a transmembrane receptor that mediates the cell-cell interactions necessary for many cell fate decisions. N has many epidermal growth factor-like repeats that are O-fucosylated by the protein O-fucosyltransferase 1 (O-Fut1), and the O-fut1 gene is essential for N signaling. However, the role of the monosaccharide O-fucose on N is unclear, because O-Fut1 also appears to have O-fucosyltransferase activity-independent functions, including as an N-specific chaperon. Such an enzymatic activity-independent function could account for the essential role of O-fut1 in N signaling. To evaluate the role of the monosaccharide O-fucose modification in N signaling, here we generated a knock-in mutant of O-fut1 (O-fut1R245A knock-in), which expresses a mutant protein that lacks O-fucosyltransferase activity but maintains the N-specific chaperon activity. Using O-fut1R245A knock-in and other gene mutations that abolish the O-fucosylation of N, we found that the monosaccharide O-fucose modification of N has a temperature-sensitive function that is essential for N signaling. The O-fucose monosaccharide and O-glucose glycan modification, catalyzed by Rumi, function redundantly in the activation of N signaling. We also showed that the redundant function of these two modifications is responsible for the presence of N at the cell surface. Our findings elucidate how different forms of glycosylation on a protein can influence the protein's functions. PMID:25378397

  13. Step-By-Step In Vitro Mutagenesis: Lessons From Fucose-Binding Lectin PA-IIL.

    Science.gov (United States)

    Mrázková, Jana; Malinovská, Lenka; Wimmerová, Michaela

    2017-01-01

    Site-directed mutagenesis is a powerful technique which is used to understand the basis of interactions between proteins and their binding partners, as well as to modify these interactions. Methods of rational design that are based on detailed knowledge of the structure of a protein of interest are often used for preliminary investigations of the possible outcomes which can result from the practical application of site-directed mutagenesis. Also, random mutagenesis can be used in tandem with site-directed mutagenesis for an examination of amino acid "hotspots."Lectins are sugar-binding proteins which, among other functions, mediate the recognition of host cells by a pathogen and its adhesion to the host cell surface. Hence, lectins and their binding properties are studied and engineered using site-directed mutagenesis.In this chapter, we describe a site-directed mutagenesis method used for investigating the sugar binding pattern of the PA-IIL lectin from the pathogenic bacterium Pseudomonas aeruginosa. Moreover, procedures for the production and purification of PA-IIL mutants are described, and several basic methods for characterizing the mutants are discussed.

  14. Measures of Malaria Burden after Long-Lasting Insecticidal Net Distribution and Indoor Residual Spraying at Three Sites in Uganda: A Prospective Observational Study

    Science.gov (United States)

    Katureebe, Agaba; Zinszer, Kate; Arinaitwe, Emmanuel; Charland, Katia; Kigozi, Ruth; Kilama, Maxwell; Nankabirwa, Joaniter; Yeka, Adoke; Mawejje, Henry; Mpimbaza, Arthur; Donnelly, Martin J.; Rosenthal, Philip J.; Lindsay, Steve W.; Staedke, Sarah G.; Smith, David L.; Kamya, Moses R.; Dorsey, Grant

    2016-01-01

    Background Long-lasting insecticidal nets (LLINs) and indoor residual spraying of insecticide (IRS) are the primary vector control interventions used to prevent malaria in Africa. Although both interventions are effective in some settings, high-quality evidence is rarely available to evaluate their effectiveness following deployment by a national malaria control program. In Uganda, we measured changes in key malaria indicators following universal LLIN distribution in three sites, with the addition of IRS at one of these sites. Methods and Findings Comprehensive malaria surveillance was conducted from October 1, 2011, to March 31, 2016, in three sub-counties with relatively low (Walukuba), moderate (Kihihi), and high transmission (Nagongera). Between 2013 and 2014, universal LLIN distribution campaigns were conducted in all sites, and in December 2014, IRS with the carbamate bendiocarb was initiated in Nagongera. High-quality surveillance evaluated malaria metrics and mosquito exposure before and after interventions through (a) enhanced health-facility-based surveillance to estimate malaria test positivity rate (TPR), expressed as the number testing positive for malaria/number tested for malaria (number of children tested for malaria: Walukuba = 42,833, Kihihi = 28,790, and Nagongera = 38,690); (b) cohort studies to estimate the incidence of malaria, expressed as the number of episodes per person-year [PPY] at risk (number of children observed: Walukuba = 340, Kihihi = 380, and Nagongera = 361); and (c) entomology surveys to estimate household-level human biting rate (HBR), expressed as the number of female Anopheles mosquitoes collected per house-night of collection (number of households observed: Walukuba = 117, Kihihi = 107, and Nagongera = 107). The LLIN distribution campaign substantially increased LLIN coverage levels at the three sites to between 65.0% and 95.5% of households with at least one LLIN. In Walukuba, over the 28-mo post-intervention period

  15. The organization of the fuc regulon specifying L-fucose dissimilation in Escherichia coli K12 as determined by gene cloning.

    Science.gov (United States)

    Chen, Y M; Zhu, Y; Lin, E C

    1987-12-01

    In Escherichia coli the six known genes specifying the utilization of L-fucose as carbon and energy source cluster at 60.2 min and constitute a regulon. These genes include fucP (encoding L-fucose permease), fucI (encoding L-fucose isomerase), fucK (encoding L-fuculose kinase), fucA (encoding L-fuculose 1-phosphate aldolase), fucO (encoding L-1,2-propanediol oxidoreductase), and fucR (encoding the regulatory protein). In this study the fuc genes were cloned and their positions on the chromosome were established by restriction endonuclease and complementation analyses. Clockwise, the gene order is: fucO-fucA-fucP-fucI-fucK-fucR. The operons comprising the structural genes and the direction of transcription were determined by complementation analysis and Southern blot hybridization. The fucPIK and fucA operons are transcribed clockwise. The fucO operon is transcribed counterclockwise. The fucR gene product activates the three structural operons in trans.

  16. Effect of colchicine on rat small intestinal absorptive cells. II. Distribution of label after incorporation of [3H]fucose into plasma membrane glycoproteins

    International Nuclear Information System (INIS)

    Ellinger, A.; Pavelka, M.; Gangl, A.

    1983-01-01

    By means of radioautography the influence was tested of various periods (5, 15, 30, 40 min, 2 hr) of pretreatment with colchicine, administered intraperitoneally to rats at a dosage of 0.5 mg/100 g of body weight, on the intracellular pathway of [ 3 H]fucose in absorptive cells of the small intestine. Administration of colchicine for 30 min and longer time intervals causes delay in the insertion of [ 3 H]fucose into the oligosaccharide chains of glycoconjugates in the Golgi apparatus, and results in redistribution of the label apparent over the different portions of the plasma membrane. In controls, at 2 and 4 hr after administration of [ 3 H]fucose the apical plasma membrane is strongly labeled. Colchicine causes equalization of the reaction of apical and basolateral regions of the plasma membrane: the number of silver grains attributable to the apical plasma membrane is reduced; following treatment with colchicine, apical portions of the plasma membrane comprise 31.6 +/- 1.8% of the silver grains, 38.6 +/- 3.8% are attributable to basolateral membrane regions. The colchicine-induced equalization of the density of label of apical and basolateral regions of the plasma membrane, in addition to the occurrence of basolateral microvillus borders, suggests microtubules to be important in the maintenance of the polar organization of small intestinal absorptive cells

  17. A real-time web-based optimal Biomass Site Assessment Tool (BioSAT): Module 1. An economic assessment of mill residues for the southern U.S.

    Science.gov (United States)

    Timothy M. Young; James H. Perdue; Andy Hartsell; Robert C. Abt; Donald Hodges; Timothy G. Rials

    2009-01-01

    Optimal locations for biomass facilities that use mill residues are identified for 13 southern U.S. states. The Biomass Site Assessment Tool (BioSAT) model is used to identify the top 20 locations for 13 southern U.S. states. The trucking cost model of BioSAT is used with Timber Mart South 2009 price data to estimate the total cost, average cost, and marginal costs for...

  18. Residual stresses

    International Nuclear Information System (INIS)

    Sahotra, I.M.

    2006-01-01

    The principal effect of unloading a material strained into the plastic range is to create a permanent set (plastic deformation), which if restricted somehow, gives rise to a system of self-balancing within the same member or reaction balanced by other members of the structure., known as residual stresses. These stresses stay there as locked-in stresses, in the body or a part of it in the absence of any external loading. Residual stresses are induced during hot-rolling and welding differential cooling, cold-forming and extruding: cold straightening and spot heating, fabrication and forced fitting of components constraining the structure to a particular geometry. The areas which cool more quickly develop residual compressive stresses, while the slower cooling areas develop residual tensile stresses, and a self-balancing or reaction balanced system of residual stresses is formed. The phenomenon of residual stresses is the most challenging in its application in surface modification techniques determining endurance mechanism against fracture and fatigue failures. This paper discusses the mechanism of residual stresses, that how the residual stresses are fanned and what their behavior is under the action of external forces. Such as in the case of a circular bar under limit torque, rectangular beam under limt moment, reclaiming of shafts welds and peening etc. (author)

  19. Optical constants and dispersion equations of lecithin, cholesterol, fucose, and chloroform: measurements in vacuum-ultraviolet to visible wavelength regions

    International Nuclear Information System (INIS)

    Andersen, M.; Nir, S.; Heller, J.M. Jr.; Painter, L.R.

    1978-01-01

    The optical constants, n and k, of solutions of lecithin, cholesterol, and fucose and of the solvent chloroform were measured for the spectral region 1348 to 6407 A by a reflectance method. Absorption peaks were found in chloroform at about 1393 and 1631 A and were attributed, respectively, to sigma → sigma/sup */ electron transitions at the C--H and C--Cl bonds and an n → sigma/sup */ electron transition at the C--Cl bond. A procedure for the determination of the optical constants of a solute from those of the solution and solvent is developed and applied. The procedure is also applied to calculate the optical constants of a solution from those of the components. From the values of the optical constants, dispersion equation parameters and van der Waals parameters were calculated for the compounds. The static electronic polarizabilities of the substances studies were found to depend little (less than 6%) on the concentration of solution used in the measurement. Values of polarizabilities obtained agreed closely with those obtained by the addition of bond polarizabilities

  20. Residual stresses

    International Nuclear Information System (INIS)

    Macherauch, E.

    1978-01-01

    Residual stresses are stresses which exist in a material without the influence of external powers and moments. They come into existence when the volume of a material constantly changes its form as a consequence of mechanical, thermal, and/or chemical processes and is hindered by neighbouring volumes. Bodies with residual stress are in mechanical balance. These residual stresses can be manifested by means of all mechanical interventions disturbing this balance. Acoustical, optical, radiological, and magnetical methods involving material changes caused by residual stress can also serve for determining residual stress. Residual stresses have an ambivalent character. In technical practice, they are feared and liked at the same time. They cause trouble because they can be the cause for unexpected behaviour of construction elements. They are feared since they can cause failure, in the worst case with catastrophical consequences. They are appreciated, on the other hand, because, in many cases, they can contribute to improvements of the material behaviour under certain circumstances. But they are especially liked for their giving convenient and (this is most important) mostly uncontrollable explanations. For only in very few cases we have enough knowledge and possibilities for the objective evaluation of residual stresses. (orig.) [de

  1. Early survival and height growth of Douglas-fir and lodgepole pine seedlings and variations in site factors following treatment of logging residues.

    Science.gov (United States)

    W. Lopushlnsky; D. Zabowskl; T.D. Anderson

    1992-01-01

    Logging residues were broadcast burned, piled and burned, removed, or left in place after clearcutting in a high-elevation subalpine fir (Abies lasiocarpa (Hook.) Nutt.) lodgepole pine (Pinus contorta Dougl. ex Loud.) forest in north-central Washington. Survival, height growth and foliar nutrient content of planted Douglas-fir...

  2. Synthesis of the oligosaccharides related to branching sites of fucosylated chondroitin sulfates from sea cucumbers.

    Science.gov (United States)

    Ustyuzhanina, Nadezhda E; Fomitskaya, Polina A; Gerbst, Alexey G; Dmitrenok, Andrey S; Nifantiev, Nikolay E

    2015-02-02

    Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. To determine pharmacophore fragments in FCS we have started systematic synthesis of oligosaccharides with well-defined structure related to various fragments of these polysaccharides. In this communication, the synthesis of non-sulfated and selectively O-sulfated di- and trisaccharides structurally related to branching sites of FCS is described. The target compounds are built up of propyl β-d-glucuronic acid residue bearing at O-3 α-l-fucosyl or α-l-fucosyl-(1→3)-α-l-fucosyl substituents. O-Sulfation pattern in the fucose units of the synthetic targets was selected according to the known to date holothurian FCS structures. Stereospecific α-glycoside bond formation was achieved using 2-O-benzyl-3,4-di-O-chloroacetyl-α-l-fucosyl trichloroacetimidate as a donor. Stereochemical outcome of the glycosylation was explained by the remote participation of the chloroacetyl groups with the formation of the stabilized glycosyl cations, which could be attacked by the glycosyl acceptor only from the α-side. The experimental results were in good agreement with the SCF/MP2 calculated energies of such participation. The synthesized oligosaccharides are regarded as model compounds for the determination of a structure-activity relationship in FCS.

  3. Analysis of myelomonocytic leukemic differentiation by a cell surface marker panel including a fucose-binding lectin from Lotus tetragonolobus.

    Science.gov (United States)

    Elias, L; Van Epps, D E

    1984-06-01

    The fucose-binding lectin from Lotus tetragonolobus ( FBL -L) has been previously shown to bind specifically to normal cells of the myeloid and monocytic lineages. The purpose of this study was to explore the utility of fluoresceinated FBL -L as a leukemia differentiation marker in conjunction with a panel of other frequently used surface markers (Fc receptor, HLA-DR, OKM1, and antimonocyte antibody). FBL -L reacted with leukemic cells in 8/9 cases of clinically recognized acute myeloid leukemia, including myeloid blast crisis of chronic granulocytic leukemia, 3/3 cases of chronic phase chronic myelogenous leukemia, and in 2/7 cases of clinically undifferentiated acute leukemia. Correlations were noted between reactivity with FBL -L, and DR and Fc receptor expression. Among continuous cell lines, FBL -L bound with high intensity to a majority of HL-60 and U937 cells. The less well differentiated myeloblast cell lines, KG-1, KG1a , and HL-60 blast II, exhibited less FBL -L binding than HL-60 and U937. A moderate proportion of K562 cells exhibited low level binding of FBL -L. Several lymphoblastic cell lines exhibited a pattern of low intensity binding that was distinguishable from the high intensity binding pattern of the myeloblastic lines. FBL -L reactivity of U937 was enhanced by induction of differentiation with leukocyte conditioned medium, but not dimethylsulfoxide. Such treatments induced contrasting patterns of change of HL-60 and U937 when labeled with OKM1, alpha-Mono, and HLA-DR. These studies demonstrate the application of FBL -L to analysis and quantitation of myelomonocytic leukemic differentiation.

  4. In silico analysis of the fucosylation-associated genome of the human blood fluke Schistosoma mansoni: cloning and characterization of the enzymes involved in GDP-L-fucose synthesis and Golgi import.

    Science.gov (United States)

    Peterson, Nathan A; Anderson, Tavis K; Wu, Xiao-Jun; Yoshino, Timothy P

    2013-07-09

    Carbohydrate structures of surface-expressed and secreted/excreted glycoconjugates of the human blood fluke Schistosoma mansoni are key determinants that mediate host-parasite interactions in both snail and mammalian hosts. Fucose is a major constituent of these immunologically important glycans, and recent studies have sought to characterize fucosylation-associated enzymes, including the Golgi-localized fucosyltransferases that catalyze the transfer of L-fucose from a GDP-L-fucose donor to an oligosaccharide acceptor. Importantly, GDP-L-fucose is the only nucleotide-sugar donor used by fucosyltransferases and its availability represents a bottleneck in fucosyl-glycotope expression. A homology-based genome-wide bioinformatics approach was used to identify and molecularly characterize the enzymes that contribute to GDP-L-fucose synthesis and Golgi import in S. mansoni. Putative functions were further investigated through molecular phylogenetic and immunocytochemical analyses. We identified homologs of GDP-D-mannose-4,6-dehydratase (GMD) and GDP-4-keto-6-deoxy-D-mannose-3,5-epimerase-4-reductase (GMER), which constitute a de novo pathway for GDP-L-fucose synthesis, in addition to a GDP-L-fucose transporter (GFT) that putatively imports cytosolic GDP-L-fucose into the Golgi. In silico primary sequence analyses identified characteristic Rossman loop and short-chain dehydrogenase/reductase motifs in GMD and GMER as well as 10 transmembrane domains in GFT. All genes are alternatively spliced, generating variants of unknown function. Observed quantitative differences in steady-state transcript levels between miracidia and primary sporocysts may contribute to differential glycotope expression in early larval development. Additionally, analyses of protein expression suggest the occurrence of cytosolic GMD and GMER in the ciliated epidermal plates and tegument of miracidia and primary sporocysts, respectively, which is consistent with previous localization of highly

  5. Field and analytical data relating to the 1972 and 1978 surveys of residual contamination of the Monte Bello Islands and Emu atomic weapons test sites

    International Nuclear Information System (INIS)

    Cooper, M.B.; Duggleby, J.C.

    1980-12-01

    Radiation surveys of the Monte Bello Islands test site in Western Australia, and the Emu test site in South Australia, were carried out in 1972 and 1978. The results have been published in ARL reports ARL/TR--010 and ARL/TR--012. The detailed field and analytical data which formed the basis of those publications are given

  6. Beta(1,2)-xylose and alpha(1,3)-fucose residues have a strong contribution in IgE binding to plant glycoallergens

    NARCIS (Netherlands)

    van Ree, R.; Cabanes-Macheteau, M.; Akkerdaas, J.; Milazzo, J. P.; Loutelier-Bourhis, C.; Rayon, C.; Villalba, M.; Koppelman, S.; Aalberse, R.; Rodriguez, R.; Faye, L.; Lerouge, P.

    2000-01-01

    Primary structures of the N-glycans of two major pollen allergens (Lol p 11 and Ole e 1) and a major peanut allergen (Ara h 1) were determined. Ole e 1 and Ara h 1 carried high mannose and complex N-glycans, whereas Lol p 11 carried only the complex. The complex structures all had a beta(1,2)-xylose

  7. Solid residues

    International Nuclear Information System (INIS)

    Mulder, E.; Duin, P.J. van; Grootenboer, G.J.

    1995-01-01

    A summary is presented of the many investigations that have been done on solid residues of atmospheric fluid bed combustion (AFBC). These residues are bed ash, cyclone ash and bag filter ash. Physical and chemical properties are discussed and then the various uses of residues (in fillers, bricks, gravel, and for recovery of aluminium) are summarised. Toxicological properties of fly ash and stack ash are discussed as are risks of pneumoconiosis for workers handling fly ash, and contamination of water by ashes. On the basis of present information it is concluded that risks to public health from exposure to emissions of coal fly ash from AFBC appear small or negligible as are health risk to workers in the coal fly ash processing industry. 35 refs., 5 figs., 12 tabs

  8. Identification of the srtC1 Transcription Start Site and Catalytically Essential Residues Required for Actinomyces oris T14V SrtC1 Activity

    Science.gov (United States)

    2011-07-27

    report the identification of the tran scription starting site of the srtC1 determined by rapid amplification of cDNA ends (RACE) method and several...When needed, kanamycin and trimethoprim were included in growth media at concentra tions of 50 and 100mg mL1, respectively. RNA isolation and...tation, resuspended in a small volume of RNase free water and stored at 80 1C. To determine the transcription start site(s) of A. oris srtC1, 50RACE PCR

  9. Fucose-Containing Sulfated Polysaccharides from Brown Seaweeds Inhibit Proliferation of Melanoma Cells and Induce Apoptosis by Activation of Caspase-3 in Vitro

    DEFF Research Database (Denmark)

    Ale, Marcel Tutor; Maruyama, Hiroko; Tamauchi, Hidekazu

    2011-01-01

    Fucose-containing sulfated polysaccharides (FCSPs) extracted from seaweeds, especially brown macro-algae, are known to possess essential bioactive properties, notably growth inhibitory effects on tumor cells. In this work, we conducted a series of in vitro studies to examine the influence of FCSPs...... of the FCSPs, particularly the presence of sulfated galactofucans (notably in S. henslowianum) and sulfated fucans (notably in F. vesiculosus). This study thus indicates that unfractionated FCSPs may exert bioactive effects on skin cancer cells via induction of apoptosis through cascades of reactions...

  10. An Examination by Site-Directed Mutagenesis of Putative Key Residues in the Determination of Coenzyme Specificity in Clostridial NAD+-Dependent Glutamate Dehydrogenase

    Directory of Open Access Journals (Sweden)

    Joanna Griffin

    2011-01-01

    Full Text Available Sequence and structure comparisons of various glutamate dehydrogenases (GDH and other nicotinamide nucleotide-dependent dehydrogenases have potentially implicated certain residues in coenzyme binding and discrimination. We have mutated key residues in Clostridium symbiosum NAD+-specific GDH to investigate their contribution to specificity and to enhance acceptance of NADPH. Comparisons with E. coli NADPH-dependent GDH prompted design of mutants F238S, P262S, and F238S/P262S, which were purified and assessed at pH 6.0, 7.0, and 8.0. They showed markedly increased catalytic efficiency with NADPH, especially at pH 8.0 (∼170-fold for P262S and F238S/P262S with relatively small changes for NADH. A positive charge introduced through the D263K mutation also greatly increased catalytic efficiency with NADPH (over 100-fold at pH 8 and slightly decreased activity with NADH. At position 242, “P6” of the “core fingerprint,” where NAD+- and NADP+-dependent enzymes normally have Gly or Ala, respectively, clostridial GDH already has Ala. Replacement with Gly produced negligible shift in coenzyme specificity.

  11. On the Formation and Properties of Interstrand DNA-DNA Cross-links Forged by Reaction of an Abasic Site With the Opposing Guanine Residue of 5′-CAp Sequences in Duplex DNA

    Science.gov (United States)

    Johnson, Kevin M.; Price, Nathan E.; Wang, Jin; Fekry, Mostafa I.; Dutta, Sanjay; Seiner, Derrick R.; Wang, Yinsheng; Gates, Kent S.

    2014-01-01

    We recently reported that the aldehyde residue of an abasic (Ap) site in duplex DNA can generate an interstrand cross-link via reaction with a guanine residue on the opposing strand. This finding is intriguing because the highly deleterious nature of interstrand cross-links suggests that even small amounts of Ap-derived cross-links could make a significant contribution to the biological consequences stemming from the generation of Ap sites in cellular DNA. Incubation of 21-bp duplexes containing a central 5′-CAp sequence under conditions of reductive amination (NaCNBH3, pH 5.2) generated much higher yields of cross-linked DNA than reported previously. At pH 7, in the absence of reducing agents, these Ap-containing duplexes also produced cross-linked duplexes that were readily detected on denaturing polyacrylamide gels. Cross-link formation was not highly sensitive to reaction conditions and, once formed, the cross-link was stable to a variety of work-up conditions. Results of multiple experiments including MALDI-TOF mass spectrometry, gel mobility, methoxyamine capping of the Ap aldehyde, inosine-for-guanine replacement, hydroxyl radical footprinting, and LCMS/MS were consistent with a cross-linking mechanism involving reversible reaction of the Ap aldehyde residue with the N2-amino group of the opposing guanine residue in 5′-CAp sequences to generate hemiaminal, imine, or cyclic hemiaminal cross-links (7-10) that were irreversibly converted under conditions of reductive amination (NaCNBH3/pH 5.2) to a stable amine linkage. Further support for the importance of the exocyclic N2-amino group in this reaction was provided by an experiment showing that installation of a 2-aminopurine-thymine base pair at the cross-linking site produced high yields (15-30%) of a cross-linked duplex at neutral pH, in the absence of NaCNBH3. PMID:23215239

  12. Extended hormone binding site of the human thyroid stimulating hormone receptor: distinctive acidic residues in the hinge region are involved in bovine thyroid stimulating hormone binding and receptor activation.

    Science.gov (United States)

    Mueller, Sandra; Kleinau, Gunnar; Jaeschke, Holger; Paschke, Ralf; Krause, Gerd

    2008-06-27

    The human thyroid stimulating hormone receptor (hTSHR) belongs to the glycoprotein hormone receptors that bind the hormones at their large extracellular domain. The extracellular hinge region of the TSHR connects the N-terminal leucine-rich repeat domain with the membrane-spanning serpentine domain. From previous studies we reasoned that apart from hormone binding at the leucine-rich repeat domain, additional multiple hormone contacts might exist at the hinge region of the TSHR by complementary charge-charge recognition. Here we investigated highly conserved charged residues in the hinge region of the TSHR by site-directed mutagenesis to identify amino acids interacting with bovine TSH (bTSH). Indeed, the residues Glu-297, Glu-303, and Asp-382 in the TSHR hinge region are essential for bTSH binding and partially for signal transduction. Side chain substitutions showed that the negative charge of Glu-297 and Asp-382 is necessary for recognition of bTSH by the hTSHR. Multiple combinations of alanine mutants of the identified positions revealed an increased negative effect on hormone binding. An assembled model suggests that the deciphered acidic residues form negatively charged patches at the hinge region resulting in an extended binding mode for bTSH on the hTSHR. Our data indicate that certain positively charged residues of bTSH might be involved in interaction with the identified negatively charged amino acids of the hTSHR hinge region. We demonstrate that the hinge region represents an extracellular intermediate connector for both hormone binding and signal transduction of the hTSHR.

  13. Residual neutron-induced radionuclides in a soil sample collected in the vicinity of the criticality accident site in Tokai-mura, Japan: A Progress Report

    International Nuclear Information System (INIS)

    Nakanishi, Takashi; Hosotani, Risa; Komura, Kazuhisa; Muroyama, Toshiharu; Kofuji, Hisaki; Murata, Yoshimasa; Kimura, Shinzo; Kumar Sahoo, Sarata; Yonehara, Hidenori; Watanabe, Yoshito; Ban-nai, Tada-aki

    2000-01-01

    Residual neutron-induced radionuclides were measured in a soil sample collected in the vicinity of the location where a criticality accident occurred (in Tokai-mura, from 30 September to 1 October, 1999). Concentrations of 24 Na, 140 La, 122 Sb, 59 Fe, 124 Sb, 46 Sc, 65 Zn, 134 Cs and 60 Co in the soil sample were determined by γ-ray spectrometry, and neutron activation analysis was carried out for selected target elements in the sample. Tentative estimates of the apparent thermal and epithermal neutron fluences which reached the sample were obtained through combined analyses of 59 Fe/ 58 Fe, 124 Sb/ 123 Sb, 46 Sc/ 45 Sc, 65 Zn/ 64 Zn, 134 Cs/ 133 Cs and 60 Co/ 59 Co

  14. Mechanism of thioredoxin-catalyzed disulfide reduction. Activation of the buried thiol and role of the variable active-site residues

    NARCIS (Netherlands)

    Carvalho, A.P.; Swart, M.; van Stralen, J.N.P.; Fernandes, P.A.; Ramos, M.E.; Bickelhaupt, F.M.

    2008-01-01

    Thioredoxins (Trx) are enzymes with a characteristic CXYC active-site motif that catalyze the reduction of disulfide bonds in other proteins. We have theoretically explored this reaction mechanism, both in the gas phase and in water, using density functional theory. The mechanism of disulfide

  15. Methodical investigations regarding the influence of narcosis on the distribution of L-3H-fucose in the brain of rats after intracerebral application

    International Nuclear Information System (INIS)

    Gutoehrle, P.

    1981-01-01

    The distribution of 3 H-fucose in the rat brain after intraventricular injection was investigated in wake and pentobarbital-anaesthetized test animals, using a method for autoradiography of water-soluble substances. By a specially developed procedure a director cannula was stereotactically implanted into a calotte through which, a few days later, an injection needle could be introduced into the lateral ventricle without narcosis. The autoradiogrammes were prepared from cryostat cuts of the cerebrum. The investigation proved the distribution of 3 H-fucose in the different sections of the brain to be very inhomogeneous and revealed that the tracer had mainly labelled the areas near the ventricle. With the aid of a micro-videomat, the blackened areas were measured in a certain frontal plane. The areas reached by the tracer, which were determined as parameters, were shown to increase as a function of the experimental time and to be always more extended in anaesthetized animals than in wake rats. (orig./MG) [de

  16. Residual basins

    International Nuclear Information System (INIS)

    D'Elboux, C.V.; Paiva, I.B.

    1980-01-01

    Exploration for uranium carried out over a major portion of the Rio Grande do Sul Shield has revealed a number of small residual basins developed along glacially eroded channels of pre-Permian age. Mineralization of uranium occurs in two distinct sedimentary units. The lower unit consists of rhythmites overlain by a sequence of black shales, siltstones and coal seams, while the upper one is dominated by sandstones of probable fluvial origin. (Author) [pt

  17. Protein and Site Specificity of Fucosylation in Liver-Secreted Glycoproteins

    Czech Academy of Sciences Publication Activity Database

    Pompach, Petr; Ashline, David J.; Brnáková, Z.; Benicky, J.; Sanda, M.; Goldman, R.

    2014-01-01

    Roč. 13, č. 12 (2014), s. 5561-5569 ISSN 1535-3893 R&D Projects: GA MŠk LH13051; GA ČR GAP206/12/0503 Grant - others:Charles Univ.(CZ) UNCE_204025/2012 Institutional support: RVO:61388971 Keywords : fucose * glycoproteins * liver * site specificity Subject RIV: CE - Biochemistry Impact factor: 4.245, year: 2014

  18. Site-Directed Mutagenesis Study Revealed Three Important Residues in Hc-DAF-22, a Key Enzyme Regulating Diapause of Haemonchus contortus

    Directory of Open Access Journals (Sweden)

    Yan Huang

    2017-11-01

    Full Text Available Haemonchus contortus (H. contortus is one of the most important parasites of small ruminants, especially goats and sheep. The complex life cycle of this nematode is a main obstacle for the control and prevention of haemonchosis. So far, a special form of arrested development called diapause different from the dauer stage in Caenorhabditis elegans (C. elegans has been found in many parasitic nematodes. In our previous study, we have characterized a novel gene Hc-daf-22 from H. contortus sharing high homology with Ce-daf-22 and functional analysis showed this gene has similar biological function with Ce-daf-22. In this study, Hc-daf-22 mutants were constructed using site-directed mutagenesis, and carried out rescue experiments, RNA interference (RNAi experiments and in vitro enzyme activity analysis with the mutants to further explore the precise function site of Hc-DAF-22. The results showed that Hc-daf-22 mutants could be expressed in the rescued ok693 worms and the expression positions were mainly in the intestine which was identical with that of Hc-daf-22 rescued worms. Through lipid staining we found that Hc-daf-22 could rescue daf-22 mutant (ok693 from the fatty acid metabolism deficiency while Hc-daf-22 mutants failed. Brood size and body length analyses in rescue experiment along with body length and life span analyses in RNAi experiment elucidated that Hc-daf-22 resembled Ce-daf-22 in effecting the development and capacity of C. elegans and mutants impaired the function of Hc-daf-22. Together with the protease activity assay, this research revealed three important active resides 84C/299H/349H in Hc-DAF-22 by site-directed mutagenesis.

  19. Site-Directed Mutagenesis Study Revealed Three Important Residues in Hc-DAF-22, a Key Enzyme Regulating Diapause of Haemonchus contortus.

    Science.gov (United States)

    Huang, Yan; Zheng, Xiuping; Zhang, Hongli; Ding, Haojie; Guo, Xiaolu; Yang, Yi; Chen, Xueqiu; Zhou, Qianjin; Du, Aifang

    2017-01-01

    Haemonchus contortus ( H. contortus ) is one of the most important parasites of small ruminants, especially goats and sheep. The complex life cycle of this nematode is a main obstacle for the control and prevention of haemonchosis. So far, a special form of arrested development called diapause different from the dauer stage in Caenorhabditis elegans ( C. elegans ) has been found in many parasitic nematodes. In our previous study, we have characterized a novel gene Hc-daf-22 from H. contortus sharing high homology with Ce-daf-22 and functional analysis showed this gene has similar biological function with Ce-daf-22 . In this study, Hc-daf-22 mutants were constructed using site-directed mutagenesis, and carried out rescue experiments, RNA interference (RNAi) experiments and in vitro enzyme activity analysis with the mutants to further explore the precise function site of Hc-DAF-22. The results showed that Hc-daf-22 mutants could be expressed in the rescued ok693 worms and the expression positions were mainly in the intestine which was identical with that of Hc-daf-22 rescued worms. Through lipid staining we found that Hc-daf-22 could rescue daf-22 mutant ( ok693 ) from the fatty acid metabolism deficiency while Hc-daf-22 mutants failed. Brood size and body length analyses in rescue experiment along with body length and life span analyses in RNAi experiment elucidated that Hc-daf-22 resembled Ce-daf-22 in effecting the development and capacity of C. elegans and mutants impaired the function of Hc-daf-22 . Together with the protease activity assay, this research revealed three important active resides 84C/299H/349H in Hc-DAF-22 by site-directed mutagenesis.

  20. H9N2 influenza virus acquires intravenous pathogenicity on the introduction of a pair of di-basic amino acid residues at the cleavage site of the hemagglutinin and consecutive passages in chickens

    Directory of Open Access Journals (Sweden)

    Sakoda Yoshihiro

    2011-02-01

    Full Text Available Abstract Background Outbreaks of avian influenza (AI caused by infection with low pathogenic H9N2 viruses have occurred in poultry, resulting in serious economic losses in Asia and the Middle East. It has been difficult to eradicate the H9N2 virus because of its low pathogenicity, frequently causing in apparent infection. It is important for the control of AI to assess whether the H9N2 virus acquires pathogenicity as H5 and H7 viruses. In the present study, we investigated whether a non-pathogenic H9N2 virus, A/chicken/Yokohama/aq-55/2001 (Y55 (H9N2, acquires pathogenicity in chickens when a pair of di-basic amino acid residues is introduced at the cleavage site of its HA molecule. Results rgY55sub (H9N2, which had four basic amino acid residues at the HA cleavage site, replicated in MDCK cells in the absence of trypsin after six consecutive passages in the air sacs of chicks, and acquired intravenous pathogenicity to chicken after four additional passages. More than 75% of chickens inoculated intravenously with the passaged virus, rgY55sub-P10 (H9N2, died, indicating that it is pathogenic comparable to that of highly pathogenic avian influenza viruses (HPAIVs defined by World Organization for Animal Health (OIE. The chickens inoculated with the virus via the intranasal route, however, survived without showing any clinical signs. On the other hand, an avirulent H5N1 strain, A/duck/Hokkaido/Vac-1/2004 (Vac1 (H5N1, acquired intranasal pathogenicity after a pair of di-basic amino acid residues was introduced into the cleavage site of the HA, followed by two passages by air sac inoculation in chicks. Conclusion The present results demonstrate that an H9N2 virus has the potential to acquire intravenous pathogenicity in chickens although the morbidity via the nasal route of infection is lower than that of H5N1 HPAIV.

  1. On-site monitoring of Hebei Spirit oil spill by fluorometric detection of oil residues in coastal waters off Taean, Korea

    Energy Technology Data Exchange (ETDEWEB)

    Kim, M.; Yim, U.H.; Hong, S.H.; Jung, J.H.; Won, J.; An, J.; Choi, H.W.; Shim, W.J. [Korea Ocean Research and Development Inst., Geoje (Korea, Republic of)

    2009-07-01

    This paper discussed activities conducted to monitor a crude oil spill that contaminated over 70 km of the western Korean coastline. Contamination levels and temporal variations of dissolved and dispersed oils in sea and pore water at 40 beaches were monitored using a portable fluorimeter for 10 months after the spill. More than 980 samples from the heavily-impacted Mallipo Beach area were analyzed. The analysis showed that oil concentrations in the sea water were as high as 16,600 {mu}g/L directly after the spill, and decreased to below the Korean marine water quality standard of 10 {mu}g/L at most sites 10 months after the spill. However, the oil content in pore water remained high, with levels of up to 2,320 {mu}g/L for the first few months following the spill. Higher oil contamination levels were observed at some sites for up to 10 months after the spill. Results of the study suggested that oil in pore water persisted in confined areas along the coastline. Results from the fluorescence detection technique were then compared with traditional gas chromatography (GC) techniques of total petroleum hydrocarbon analysis. It was concluded that fluorescence detection was capable of generating accurate results more quickly and cost-effectively than traditional GC techniques. 22 refs., 7 figs.

  2. Mapping Sites of O-Glycosylation and Fringe Elongation on Drosophila Notch*

    Science.gov (United States)

    Harvey, Beth M.; Rana, Nadia A.; Moss, Hillary; Leonardi, Jessica; Jafar-Nejad, Hamed; Haltiwanger, Robert S.

    2016-01-01

    Glycosylation of the Notch receptor is essential for its activity and serves as an important modulator of signaling. Three major forms of O-glycosylation are predicted to occur at consensus sites within the epidermal growth factor-like repeats in the extracellular domain of the receptor: O-fucosylation, O-glucosylation, and O-GlcNAcylation. We have performed comprehensive mass spectral analyses of these three types of O-glycosylation on Drosophila Notch produced in S2 cells and identified peptides containing all 22 predicted O-fucose sites, all 18 predicted O-glucose sites, and all 18 putative O-GlcNAc sites. Using semiquantitative mass spectral methods, we have evaluated the occupancy and relative amounts of glycans at each site. The majority of the O-fucose sites were modified to high stoichiometries. Upon expression of the β3-N-acetylglucosaminyltransferase Fringe with Notch, we observed varying degrees of elongation beyond O-fucose monosaccharide, indicating that Fringe preferentially modifies certain sites more than others. Rumi modified O-glucose sites to high stoichiometries, although elongation of the O-glucose was site-specific. Although the current putative consensus sequence for O-GlcNAcylation predicts 18 O-GlcNAc sites on Notch, we only observed apparent O-GlcNAc modification at five sites. In addition, we performed mass spectral analysis on endogenous Notch purified from Drosophila embryos and found that the glycosylation states were similar to those found on Notch from S2 cells. These data provide foundational information for future studies investigating the mechanisms of how O-glycosylation regulates Notch activity. PMID:27268051

  3. Important Determinants for Fucoidan Bioactivity: A Critical Review of Structure-Function Relations and Extraction Methods for Fucose-Containing Sulfated Polysaccharides from Brown Seaweeds

    DEFF Research Database (Denmark)

    Ale, Marcel Tutor; Mikkelsen, Jørn Dalgaard; Meyer, Anne S.

    2011-01-01

    Seaweeds—or marine macroalgae—notably brown seaweeds in the class Phaeophyceae, contain fucoidan. Fucoidan designates a group of certain fucose-containing sulfated polysaccharides (FCSPs) that have a backbone built of (1→3)-linked α-l-fucopyranosyl or of alternating (1→3)- and (1→4)-linked α...... for humans. The bioactive properties may vary depending on the source of seaweed, the compositional and structural traits, the content (charge density), distribution, and bonding of the sulfate substitutions, and the purity of the FCSP product. The preservation of the structural integrity of the FCSP...... on the most recent developments in the chemistry of fucoidan/FCSPs emphasizing the significance of different extraction techniques for the structural composition and biological activity with particular focus on sulfate groups....

  4. Residual nilpotence and residual solubility of groups

    International Nuclear Information System (INIS)

    Mikhailov, R V

    2005-01-01

    The properties of the residual nilpotence and the residual solubility of groups are studied. The main objects under investigation are the class of residually nilpotent groups such that each central extension of these groups is also residually nilpotent and the class of residually soluble groups such that each Abelian extension of these groups is residually soluble. Various examples of groups not belonging to these classes are constructed by homological methods and methods of the theory of modules over group rings. Several applications of the theory under consideration are presented and problems concerning the residual nilpotence of one-relator groups are considered.

  5. Size distributions of non-volatile particle residuals (Dp<800 nm at a rural site in Germany and relation to air mass origin

    Directory of Open Access Journals (Sweden)

    T. Tuch

    2007-11-01

    Full Text Available Atmospheric aerosol particle size distributions at a continental background site in Eastern Germany were examined for a one-year period. Particles were classified using a twin differential mobility particle sizer in a size range between 3 and 800 nm. As a novelty, every second measurement of this experiment involved the removal of volatile chemical compounds in a thermodenuder at 300°C. This concept allowed to quantify the number size distribution of non-volatile particle cores – primarily associated with elemental carbon, and to compare this to the original non-conditioned size distribution. As a byproduct of the volatility analysis, new particles originating from nucleation inside the thermodenuder can be observed, however, overwhelmingly at diameters below 6 nm. Within the measurement uncertainty, every particle down to particle sizes of 15 nm is concluded to contain a non-volatile core. The volume fraction of non-volatile particulate matter (non-conditioned diameter < 800 nm varied between 10 and 30% and was largely consistent with the experimentally determined mass fraction of elemental carbon. The average size of the non-volatile particle cores was estimated as a function of original non-conditioned size using a summation method, which showed that larger particles (>200 nm contained more non-volatile compounds than smaller particles (<50 nm, thus indicating a significantly different chemical composition. Two alternative air mass classification schemes based on either, synoptic chart analysis (Berliner Wetterkarte or back trajectories showed that the volume and number fraction of non-volatile cores depended less on air mass than the total particle number concentration. In all air masses, the non-volatile size distributions showed a more and a less volatile ("soot" mode, the latter being located at about 50 nm. During unstable conditions and in maritime air masses, smaller values were observed compared to stable or continental conditions

  6. Two-magnetization Nordheim model of randomly distributed Co local sites for the anomalous residual resistivity at the magnetic phase boundary of Y1-xRxCo2 system (R: rare earth)

    International Nuclear Information System (INIS)

    Yagasaki, K; Nakama, T; Takaesu, Y; Hedo, M; Uchima, K; Uwatoko, Y; Burkov, A

    2009-01-01

    The electrical resistivity ν of the Laves phase Y 1-x R x Co 2 compound system has been measured in magnetic fields up to 10 T and under pressures up to 8 GPa at temperatures from 1.5 to 300 K. The anomalous behavior of residual resistivity has been observed in a region x a , where x a is a critical concentration between inhomogeneously and homogeneously ordered phases, and which has a maximum at x c where T c ∼ 0 with a mean field acting on Co sub-lattice is equal to the itinerant Co metamagnetic critical field B c . In x c a , the magneto-resistivity and pressure resistivity are anomalously large with positive sign. However, in the paramagnetic region for x c , they are anomalously large but with negative sign. The anomalous behavior is attributed to the s-d scattering of conduction electrons due to statistically disordered Co magnetization. Those phenomena can be explained by a new scattering model of [Two magnetization Nordheim model for randomly distributed Co sites] introduced by us.

  7. Altered metabolic incorporation of fucose and leucine into PNS myelin of 25-week-old diabetic (C57BL/Ks [db/db]) mice: effects of untreated diabetes on nerve metabolism

    International Nuclear Information System (INIS)

    Chez, M.G.; Peterson, R.G.

    1983-01-01

    Sciatic nerves of 25-week-old genetically diabetic (C57BL/Ks [db/db]) mice and their litter-mate controls were removed, and their metabolic incorporation of [ 3 H]fucose and [ 14 C]leucine into myelin was studied in vitro. Untreated diabetic animals showed significant increases (p less than 0.05) in the fucose/leucine incorporation into myelin when compared to values found for their litter-mates. These results correlated well with previous experiments performed on alloxan or streptozotocin-diabetic rats and thus show the in vitro incubation procedure to be a good indicator of altered metabolic conditions in peripheral nerves due to diabetes mellitus. The resulting ratio increases seen in diabetic animals is at variance with the decrease in ratios found in animals undergoing typical Wallerian degeneration. These results suggest that different metabolic processes operate in untreated diabetics than in normals or in those undergoing other degenerative nerve processes

  8. A characteristic chondroitin sulfate trisaccharide unit with a sulfated fucose branch exhibits neurite outgrowth-promoting activity: Novel biological roles of fucosylated chondroitin sulfates isolated from the sea cucumber Apostichopus japonicus.

    Science.gov (United States)

    Shida, Miharu; Mikami, Tadahisa; Tamura, Jun-Ichi; Kitagawa, Hiroshi

    2017-06-03

    Chondroitin sulfate (CS) is a class of sulfated glycosaminoglycan (GAG) chains that consist of repeating disaccharide unit composed of glucuronic acid (GlcA) and N-acetylgalactosamine (GalNAc). CS chains are found throughout the pericellular and extracellular spaces and contribute to the formation of functional microenvironments for numerous biological events. However, their structure-function relations remain to be fully characterized. Here, a fucosylated CS (FCS) was isolated from the body wall of the sea cucumber Apostichopus japonicus. Its promotional effects on neurite outgrowth were assessed by using isolated polysaccharides and the chemically synthesized FCS trisaccharide β-D-GalNAc(4,6-O-disulfate) (1-4)[α-l-fucose (2,4-O-disulfate) (1-3)]-β-D-GlcA. FCS polysaccharides contained the E-type disaccharide unit GlcA-GalNAc(4,6-O-disulfate) as a CS major backbone structure and carried distinct sulfated fucose branches. Despite their relatively lower abundance of E unit, FCS polysaccharides exhibited neurite outgrowth-promoting activity comparable to squid cartilage-derived CS-E polysaccharides, which are characterized by their predominant E units, suggesting potential roles of the fucose branch in neurite outgrowth. Indeed, the chemically synthesized FCS trisaccharide was as effective as CS-E tetrasaccharide in stimulating neurite elongation in vitro. In conclusion, FCS trisaccharide units with 2,4-O-disulfated fucose branches may provide new insights into understanding the structure-function relations of CS chains. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. F-Type Lectins: A Highly Diversified Family of Fucose-Binding Proteins with a Unique Sequence Motif and Structural Fold, Involved in Self/Non-Self-Recognition

    Directory of Open Access Journals (Sweden)

    Gerardo R. Vasta

    2017-11-01

    Full Text Available The F-type lectin (FTL family is one of the most recent to be identified and structurally characterized. Members of the FTL family are characterized by a fucose recognition domain [F-type lectin domain (FTLD] that displays a novel jellyroll fold (“F-type” fold and unique carbohydrate- and calcium-binding sequence motifs. This novel lectin family comprises widely distributed proteins exhibiting single, double, or greater multiples of the FTLD, either tandemly arrayed or combined with other structurally and functionally distinct domains, yielding lectin subunits of pleiotropic properties even within a single species. Furthermore, the extraordinary variability of FTL sequences (isoforms that are expressed in a single individual has revealed genetic mechanisms of diversification in ligand recognition that are unique to FTLs. Functions of FTLs in self/non-self-recognition include innate immunity, fertilization, microbial adhesion, and pathogenesis, among others. In addition, although the F-type fold is distinctive for FTLs, a structure-based search revealed apparently unrelated proteins with minor sequence similarity to FTLs that displayed the FTLD fold. In general, the phylogenetic analysis of FTLD sequences from viruses to mammals reveals clades that are consistent with the currently accepted taxonomy of extant species. However, the surprisingly discontinuous distribution of FTLDs within each taxonomic category suggests not only an extensive structural/functional diversification of the FTLs along evolutionary lineages but also that this intriguing lectin family has been subject to frequent gene duplication, secondary loss, lateral transfer, and functional co-option.

  10. Fate and role of macromolecules synthesized during mammalian oocyte meiotic maturation. II. - Autoradiographic topography of (/sup 3/H)-fucose incorporation in pig oocytes cultured in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Pivko, J. (Animal Production Research Institute, Nitra (Czechoslovakia)); Motlik, J. (Institute of Animal Physiology and Genetics, Libechov (Czechoslovakia)); Kopecny, V. (University J.E. Purkyne, Brno (Czechoslovakia)); Flechon, J.E. (I.N.R.A., Station Centrale de Physiologie Animale, Jouy-en-Josas (France))

    1982-01-01

    Pig oocytes in different maturational stages--germinal vesicle (GV), metaphase I (MI) and metaphase II (MII)- were cultured in vitro with (/sup 3/H)-fucose. The incorporation of the precursor was followed by LM or EM autoradiography on air-dried preparations and on semithin or thin sections. The cumulus cells connected with oocytes at the GV stage were intensely labelled, while the labelling of the cumulus of MI and MII oocytes was lower. The cytoplasm of oocytes in the GV stage was characterized by nests of silver grains located mainly in a juxtanuclear position. The accumulation of label in the cortical region, observed in oocytes cultured with an intact cumulus, was less evident in cumulus-deprived oocytes. Lower labelling of the ooplasm, together with uniform distribution of the grains, was observed in later stages of meiosis. EM autoradiographs demonstrated the main localization, at the GV stage, of label in the Golgi apparatus and near the cell surface of oocytes and cumulus cells, as well as in the cytoplasmic processes of corona radiata cells. It is concluded that a relatively intense glycoprotein synthesis takes place in pig oocytes and cumulus cells during resumption of meiosis, at least before GV breakdown. Metabolic cooperation may occur as long as oocytes and cumulus cells keep membrane junctions.

  11. Nonencapsulated or nontypeable Haemophilus influenzae are more likely than their encapsulated or serotypeable counterparts to have mutations in their fucose operon.

    Science.gov (United States)

    Shuel, Michelle L; Karlowsky, Kathleen E; Law, Dennis K S; Tsang, Raymond S W

    2011-12-01

    Population biology of Haemophilus influenzae can be studied by multilocus sequence typing (MLST), and isolates are assigned sequence types (STs) based on nucleotide sequence variations in seven housekeeping genes, including fucK. However, the ST cannot be assigned if one of the housekeeping genes is absent or cannot be detected by the current protocol. Occasionally, strains of H. influenzae have been reported to lack the fucK gene. In this study, we examined the prevalence of this mutation among our collection of H. influenzae isolates. Of the 704 isolates studied, including 282 encapsulated and 422 nonencapsulated isolates, nine were not typeable by MLST owing to failure to detect the fucK gene. All nine fucK-negative isolates were nonencapsulated and belonged to various biotypes. DNA sequencing of the fucose operon region confirmed complete deletion of genes in the operon in seven of the nine isolates, while in the remaining two isolates, some of the genes were found intact or in parts. The significance of these findings is discussed.

  12. Presence of fucosyl residues on the oligosaccharide antennae of membrane glycopeptides of human neuroblastoma cells

    International Nuclear Information System (INIS)

    Santer, U.V.; Glick, M.C.

    1983-01-01

    Fucosyl residues linked alpha 1 leads to 3 or 4 to N-acetylglucosamine were found in large amounts on glycopeptides from the membranes of human tumor cells of neurectodermal origin but not on membrane glycopeptides from human fibroblasts. The fucosyl residues were detected by release of radioactive fucose from the glycopeptides with an almond alpha-L-fucosidase specific for fucosyl alpha 1 leads to 3(4)-N-acetylglucosamine. In other studies, the linkage was shown to be alpha 1 leads to 3 by nuclear magnetic resonance analysis. Glycopeptides containing these fucosyl residues from four human neuroblastoma cell lines were defined by binding to immobilized lectins. In addition, the glycopeptides from one human neuroblastoma cell line, CHP-134, were further characterized by enzyme degradation and columns calibrated for size and charge. The antennary position of fucosyl alpha 1 leads to 3-N-acetylglucosamine on the glycopeptides was demonstrated by the use of exoglycosidases and endoglycosidase D, since complete degradation to yield fucosyl-N-acetylglucosaminylasparagine was obtained only after treatment with almond alpha-L-fucosidase prior to the sequential degradation. Fucosyl alpha 1 leads to 3-N-acetylglucosamine was present on most size and charge classes of membrane glycopeptides and therefore was not limited to a few glycoproteins. Since the almond alpha-L-fucosidase cleaves fucosyl residues from glycoproteins, the physiological effects of the increased specific fucosylation on human tumors of neurectodermal origin can be examined

  13. Using cotton plant residue to produce briquettes

    Energy Technology Data Exchange (ETDEWEB)

    Coates, W. [University of Arizona, Tucson, AZ (United States). Bioresources Research Facility

    2000-07-01

    In Arizona, cotton (Gossypium) plant residue left in the field following harvest must be buried to prevent it from serving as an overwintering site for insects such as the pink bollworm. Most tillage operations employed to incorporate the residue into the soil are energy intensive and often degrade soil structure. Trials showed that cotton plant residue could be incorporated with pecan shells to produce commercially acceptable briquettes. Pecan shell briquettes containing cotton residue rather than waste paper were slightly less durable, when made using equivalent weight mixtures and moisture contents. Proximate and ultimate analyses showed the only difference among briquette samples to be a higher ash content in those made using cotton plant residue. Briquettes made with paper demonstrated longer flame out time, and lower ash percentage, compared to those made with cotton plant residue. (author)

  14. Minimization of zirconium chlorinator residues

    International Nuclear Information System (INIS)

    Green, G.K.; Harbuck, D.D.

    1995-01-01

    Zirconium chlorinator residues contain an array of rare earths, scandium, unreacted coke, and radioactive thorium and radium. Because of the radioactivity, the residues must be disposed in special waste containment facilities. As these sites become more congested, and with stricter environmental regulations, disposal of large volumes of wastes may become more difficult. To reduce the mass of disposed material, the US Bureau of Mines (USBM) developed technology to recover rare earths, thorium and radium, and unreacted coke from these residues. This technology employs an HCl leach to solubilize over 99% of the scandium and thorium, and over 90% of the rare earths. The leach liquor is processed through several solvent extraction stages to selectively recover scandium, thorium, and rare earths. The leach residue is further leached with an organic acid to solubilize radium, thus allowing unreacted coke to be recycled to the chlorinator. The thorium and radium waste products, which comprise only 2.1% of the original residue mass, can then be sent to the radioactive waste facility

  15. Residue management at Rocky Flats

    International Nuclear Information System (INIS)

    Olencz, J.

    1995-01-01

    Past plutonium production and manufacturing operations conducted at the Rocky Flats Environmental Technology Site (RFETS) produced a variety of plutonium-contaminated by-product materials. Residues are a category of these materials and were categorized as open-quotes materials in-processclose quotes to be recovered due to their inherent plutonium concentrations. In 1989 all RFETS plutonium production and manufacturing operations were curtailed. This report describes the management of plutonium bearing liquid and solid wastes

  16. Histochemistry of lectin-binding sites in Halicryptus spinulosus (Priapulida).

    Science.gov (United States)

    Busch, A; Schumacher, U; Storch, V

    2001-02-01

    Priapulida represent one of the phylogenetically oldest multicellular animal groups. In multicellular animals (Metazoa) cell-to-cell and cell-to-matrix interactions are often mediated by carbohydrate residues of glycoconjugates. To analyze the carbohydrate composition of a phylogenetically old species, lectin histochemistry was employed on 5 specimens of the priapulid Halicryptus spinulosus. Many lectins bound to the chitin-containing cuticle, including those specific for carbohydrates other than N-acetylglucosamine, the principle building block of chitin. The connective tissue of the animals contained both N-acetylglucosamine and N-acetylgalactosamine. Mannose residues were widely distributed with the exception of the cuticle, but complex type carbohydrates were not present in the entire animal. Sialic acid residues were only detected in the cuticle and brush border of the intestinal epithelium, while fucose was limited to the cuticle. Thus, the lectin-binding pattern indicated that sugars typical for the linking region of both N- and O-glycoproteins in mammals are also present in H. spinulosus. Carbohydrate residues that are typical for the complex type of N-linked glycans in vertebrates are not present as are carbohydrate residues typical for the termination of O-linked carbohydrate chains. Hence, a truncated form of both N- and O-linked glycosylation is present in H. spinulosus indicating that more complex patterns of glycosylation developed later during evolution.

  17. Stabilization of plutonium bearing residues at Lawrence Livermore National Laboratory

    International Nuclear Information System (INIS)

    Bronson, M.C.; Van Konynenburg, R.A.; Ebbinghaus, B.B.

    1995-01-01

    The US Department of Energy's (US DOE) Lawrence Livermore National Laboratory (LLNL) has plutonium holdings including metal, oxide and residue materials, all of which need stabilization of some type. Residue materials include calcined ash, calcined precipitates, pyrochemical salts, glove box sweepings, metallurgical samples, graphite, and pyrochemical ceramic crucibles. These residues are typical of residues stored throughout the US DOE plutonium sites. The stabilization process selected for each of these residues requires data on chemical impurities, physical attributes, and chemical forms of the plutonium. This paper outlines the characterization and stabilization of LLNL ash residues, pyrochemical salts, and graphite

  18. Calcination/dissolution residue treatment

    International Nuclear Information System (INIS)

    Knight, R.C.; Creed, R.F.; Patello, G.K.; Hollenberg, G.W.; Buehler, M.F.; O'Rourke, S.M.; Visnapuu, A.; McLaughlin, D.F.

    1994-09-01

    Currently, high-level wastes are stored underground in steel-lined tanks at the Hanford site. Current plans call for the chemical pretreatment of these wastes before their immobilization in stable glass waste forms. One candidate pretreatment approach, calcination/dissolution, performs an alkaline fusion of the waste and creates a high-level/low-level partition based on the aqueous solubilities of the components of the product calcine. Literature and laboratory studies were conducted with the goal of finding a residue treatment technology that would decrease the quantity of high-level waste glass required following calcination/dissolution waste processing. Four elements, Fe, Ni, Bi, and U, postulated to be present in the high-level residue fraction were identified as being key to the quantity of high-level glass formed. Laboratory tests of the candidate technologies with simulant high-level residues showed reductive roasting followed by carbonyl volatilization to be successful in removing Fe, Ni, and Bi. Subsequent bench-scale tests on residues from calcination/dissolution processing of genuine Hanford Site tank waste showed Fe was separated with radioelement decontamination factors of 70 to 1,000 times with respect to total alpha activity. Thermodynamic analyses of the calcination of five typical Hanford Site tank waste compositions also were performed. The analyses showed sodium hydroxide to be the sole molten component in the waste calcine and emphasized the requirement for waste blending if fluid calcines are to be achieved. Other calcine phases identified in the thermodynamic analysis indicate the significant thermal reconstitution accomplished in calcination

  19. Mutational properties of amino acid residues: implications for evolvability of phosphorylatable residues

    DEFF Research Database (Denmark)

    Creixell, Pau; Schoof, Erwin M.; Tan, Chris Soon Heng

    2012-01-01

    in terms of their mutational activity. Moreover, we highlight the importance of the genetic code and physico-chemical properties of the amino acid residues as likely causes of these inequalities and uncover serine as a mutational hot spot. Finally, we explore the consequences that these different......; it is typically assumed that all amino acid residues are equally likely to mutate or to result from a mutation. Here, by reconstructing ancestral sequences and computing mutational probabilities for all the amino acid residues, we refute this assumption and show extensive inequalities between different residues...... mutational properties have on phosphorylation site evolution, showing that a higher degree of evolvability exists for phosphorylated threonine and, to a lesser extent, serine in comparison with tyrosine residues. As exemplified by the suppression of serine's mutational activity in phosphorylation sites, our...

  20. Residual gas analysis

    International Nuclear Information System (INIS)

    Berecz, I.

    1982-01-01

    Determination of the residual gas composition in vacuum systems by a special mass spectrometric method was presented. The quadrupole mass spectrometer (QMS) and its application in thin film technology was discussed. Results, partial pressure versus time curves as well as the line spectra of the residual gases in case of the vaporization of a Ti-Pd-Au alloy were demonstrated together with the possible construction schemes of QMS residual gas analysers. (Sz.J.)

  1. 5,5'-Dithiobis-(2-nitrobenzoic acid) as a probe for a non-essential cysteine residue at the medium chain acyl-coenzyme A dehydrogenase binding site of the human 'electron transferring flavoprotein' (ETF).

    Science.gov (United States)

    Parker, A; Engel, P C

    1999-01-01

    Human 'electron transferring flavoprotein' (ETF) was inactivated by the thiol-specific reagent 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB). The kinetic profile showed the reaction followed pseudo-first-order kinetics during the initial phase of inactivation. Monitoring the release of 5-thio-2-nitrobenzoate (TNB) showed that modification of 1 cysteine residue was responsible for the loss of activity. The inactivation of ETF by DTNB could be reversed upon incubation with thiol-containing reagents. The loss of activity was prevented by the inclusion of medium chain acyl-CoA dehydrogenase (MCAD) and octanoyl-CoA. Cyanolysis of the DTNB modified-ETF with KCN led to the release of TNB accompanied presumably by the formation of the thio-cyano enzyme and with almost full recovery of activity. Conservation studies and the lack of 100% inactivation, however, suggested that this cysteine residue is not essential for the interaction with MCAD.

  2. Logging and Agricultural Residue Supply Curves for the Pacific Northwest

    Energy Technology Data Exchange (ETDEWEB)

    Kerstetter, James D.; Lyons, John Kim

    2001-01-01

    This report quantified the volume of logging residues at the county level for current timber harvests. The cost of recovering logging residues was determined for skidding, yearding, loading, chipping and transporting the residues. Supply curves were developed for ten candidate conversion sites in the Pacific Northwest Region. Agricultural field residues were also quantified at the county level using five-year average crop yields. Agronomic constraints were applied to arrive at the volumes available for energy use. Collection costs and transportation costs were determined and supply curves generated for thirteen candidate conversion sites.

  3. Handling of Solid Residues

    International Nuclear Information System (INIS)

    Medina Bermudez, Clara Ines

    1999-01-01

    The topic of solid residues is specifically of great interest and concern for the authorities, institutions and community that identify in them a true threat against the human health and the atmosphere in the related with the aesthetic deterioration of the urban centers and of the natural landscape; in the proliferation of vectorial transmitters of illnesses and the effect on the biodiversity. Inside the wide spectrum of topics that they keep relationship with the environmental protection, the inadequate handling of solid residues and residues dangerous squatter an important line in the definition of political and practical environmentally sustainable. The industrial development and the population's growth have originated a continuous increase in the production of solid residues; of equal it forms, their composition day after day is more heterogeneous. The base for the good handling includes the appropriate intervention of the different stages of an integral administration of residues, which include the separation in the source, the gathering, the handling, the use, treatment, final disposition and the institutional organization of the administration. The topic of the dangerous residues generates more expectation. These residues understand from those of pathogen type that are generated in the establishments of health that of hospital attention, until those of combustible, inflammable type, explosive, radio-active, volatile, corrosive, reagent or toxic, associated to numerous industrial processes, common in our countries in development

  4. Sustainable System for Residual Hazards Management

    International Nuclear Information System (INIS)

    Kevin M. Kostelnik; James H. Clarke; Jerry L. Harbour

    2004-01-01

    Hazardous, radioactive and other toxic substances have routinely been generated and subsequently disposed of in the shallow subsurface throughout the world. Many of today's waste management techniques do not eliminate the problem, but rather only concentrate or contain the hazardous contaminants. Residual hazards result from the presence of hazardous and/or contaminated material that remains on-site following active operations or the completion of remedial actions. Residual hazards pose continued risk to humans and the environment and represent a significant and chronic problem that require continuous long-term management (i.e. >1000 years). To protect human health and safeguard the natural environment, a sustainable system is required for the proper management of residual hazards. A sustainable system for the management of residual hazards will require the integration of engineered, institutional and land-use controls to isolate residual contaminants and thus minimize the associated hazards. Engineered controls are physical modifications to the natural setting and ecosystem, including the site, facility, and/or the residual materials themselves, in order to reduce or eliminate the potential for exposure to contaminants of concern (COCs). Institutional controls are processes, instruments, and mechanisms designed to influence human behavior and activity. System failure can involve hazardous material escaping from the confinement because of system degradation (i.e., chronic or acute degradation) or by external intrusion of the biosphere into the contaminated material because of the loss of institutional control. An ongoing analysis of contemporary and historic sites suggests that the significance of the loss of institutional controls is a critical pathway because decisions made during the operations/remedial action phase, as well as decisions made throughout the residual hazards management period, are key to the long-term success of the prescribed system. In fact

  5. Environmental dredging residual generation and management.

    Science.gov (United States)

    Patmont, Clay; LaRosa, Paul; Narayanan, Raghav; Forrest, Casey

    2018-05-01

    The presence and magnitude of sediment contamination remaining in a completed dredge area can often dictate the success of an environmental dredging project. The need to better understand and manage this remaining contamination, referred to as "postdredging residuals," has increasingly been recognized by practitioners and investigators. Based on recent dredging projects with robust characterization programs, it is now understood that the residual contamination layer in the postdredging sediment comprises a mixture of contaminated sediments that originate from throughout the dredge cut. This mixture of contaminated sediments initially exhibits fluid mud properties that can contribute to sediment transport and contamination risk outside of the dredge area. This article reviews robust dredging residual evaluations recently performed in the United States and Canada, including the Hudson River, Lower Fox River, Ashtabula River, and Esquimalt Harbour, along with other projects. These data better inform the understanding of residuals generation, leading to improved models of dredging residual formation to inform remedy evaluation, selection, design, and implementation. Data from these projects confirm that the magnitude of dredging residuals is largely determined by site conditions, primarily in situ sediment fluidity or liquidity as measured by dry bulk density. While the generation of dredging residuals cannot be avoided, residuals can be successfully and efficiently managed through careful development and implementation of site-specific management plans. Integr Environ Assess Manag 2018;14:335-343. © 2018 The Authors. Integrated Environmental Assessment and Management Published by Wiley Periodicals, Inc. on behalf of Society of Environmental Toxicology & Chemistry (SETAC). © 2018 The Authors. Integrated Environmental Assessment and Management Published by Wiley Periodicals, Inc. on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

  6. A manual for implementing residual radioactive material guidelines

    International Nuclear Information System (INIS)

    Gilbert, T.L.; Yu, C.; Yuan, Y.C.; Zielen, A.J.; Jusko, M.J.; Wallo, A. III; Argonne National Lab., IL; Dames and Moore, West Valley, NY; Argonne National Lab., IL; USDOE Assistant Secretary for Nuclear Energy, Washington, DC

    1989-06-01

    This manual presents information for implementing US Department of Energy (DOE) guidelines for residual radioactive material at sites identified by the Formerly Utilized Sites Remedial Action Program (FUSRAP) and the Surplus Facilities Management Program (SFMP). It describes the analysis and models used to derive site-specific guidelines for allowable residual concentrations of radionuclides in soil and the design and use of the RESRAD computer code for calculating guideline values. It also describes procedures for implementing DOE policy for reducing residual radioactivity to levels that are as low as reasonably achievable. 36 refs., 16 figs, 22 tabs

  7. Contaminant transport at a waste residue deposit

    DEFF Research Database (Denmark)

    Engesgaard, Peter Knudegaard; Traberg, Rikke

    1996-01-01

    Contaminant transport in an aquifer at an incinerator waste residue deposit in Denmark is simulated. A two-dimensional, geochemical transport code is developed for this purpose and tested by comparison to results from another code, The code is applied to a column experiment and to the field site...

  8. 76 FR 42074 - Consideration of Rulemaking To Address Prompt Remediation of Residual Radioactivity During...

    Science.gov (United States)

    2011-07-18

    ... Address Prompt Remediation of Residual Radioactivity During Operations AGENCY: Nuclear Regulatory... radioactivity during the operational phase of licensed material sites and nuclear reactors. The NRC has not... to the decommissioning planning process by addressing remediation of residual radioactivity during...

  9. Gold processing residue from Jacobina Basin: chemical and physical properties

    OpenAIRE

    Lima, Luiz Rogério Pinho de Andrade; Bernardez, Letícia Alonso; Barbosa, Luís Alberto Dantas

    2007-01-01

    p. 848-852 Gold processing residues or tailings are found in several areas in the Itapicuru River region (Bahia, Brazil), and previous studies indicated significant heavy metals content in the river sediments. The present work focused on an artisanal gold processing residue found in a site from this region. Samples were taken from the processing residue heaps and used to perform a physical and chemical characterization study using X-ray diffraction, scanning electron microscopy, neutron...

  10. DEVISING STRATEGIES FOR RECLAMATION OF DERELICT SITES DUE TO MINING OF RESIDUAL SOIL (“SAIBRO” AT UBATUBA, NORTH COAST OF SÃO PAULO STATE, BRAZIL: THE VIEWS AND ROLES OF THE STAKEHOLDERS

    Directory of Open Access Journals (Sweden)

    Cláudio José Ferreira

    2005-05-01

    Full Text Available Between the late 60's and early 90's intensive exploitation of residual soil for civil construction took place in the municipality of Ubatuba, North Coast of São Paulo State. Tropical climatic conditions associated with a hilly landscape bordered by the Atlantic Ocean gave rise to thick (10-30m weathering profiles derived from Precambrian granitic-gneissic rocks. Exploitation of large volumes of this material required only very simple technology, which on the other hand has caused highly adverse environmental impacts, such asdeforestation, soil erosion, land instability hazards, scenic eterioration, pollution and disturbance of local water flow regime. This paper deals with the devising of strategies thus identifying stakeholders and describing their roles both on the dereliction and reclamation processes related with residual soil exploitation. The evaluation approach focused on the analysis of the regulatory framework and its practical implications and issues in terms of licensing for mining operations, commercialization of construction materials, and the relations of such mining activities with regional and urban planning. A survey of stakeholderviewpoints was successfully achieved through a two-day workshop. The attendance included decision-makers, officials and representatives of governmental and regulatory bodies (Federal, State and Municipal, environmental research institutions, private miners (entrepreneurs and consultants, public attorney, and NGOs. The outcomes have ndicated that State and local authorities properly managed to stop illegal exploitation of residual soil in the early 90´s. However no land reclamation was undertaken until present. One of the main problems would be related with the procedures for mining and environmental licensing that were considered to be too complex and long (usually involving up to seven decisional instances and 2,5 years in average. Other key issues and needs identified included: aorganization of a

  11. Characterization of Hospital Residuals

    International Nuclear Information System (INIS)

    Blanco Meza, A.; Bonilla Jimenez, S.

    1997-01-01

    The main objective of this investigation is the characterization of the solid residuals. A description of the handling of the liquid and gassy waste generated in hospitals is also given, identifying the source where they originate. To achieve the proposed objective the work was divided in three stages: The first one was the planning and the coordination with each hospital center, in this way, to determine the schedule of gathering of the waste can be possible. In the second stage a fieldwork was made; it consisted in gathering the quantitative and qualitative information of the general state of the handling of residuals. In the third and last stage, the information previously obtained was organized to express the results as the production rate per day by bed, generation of solid residuals for sampled services, type of solid residuals and density of the same ones. With the obtained results, approaches are settled down to either determine design parameters for final disposition whether for incineration, trituration, sanitary filler or recycling of some materials, and storage politics of the solid residuals that allow to determine the gathering frequency. The study concludes that it is necessary to improve the conditions of the residuals handling in some aspects, to provide the cleaning personnel of the equipment for gathering disposition and of security, minimum to carry out this work efficiently, and to maintain a control of all the dangerous waste, like sharp or polluted materials. In this way, an appreciable reduction is guaranteed in the impact on the atmosphere. (Author) [es

  12. A putative carbohydrate-binding domain of the lactose-binding Cytisus sessilifolius anti-H(O) lectin has a similar amino acid sequence to that of the L-fucose-binding Ulex europaeus anti-H(O) lectin.

    Science.gov (United States)

    Konami, Y; Yamamoto, K; Osawa, T; Irimura, T

    1995-04-01

    The complete amino acid sequence of a lactose-binding Cytisus sessilifolius anti-H(O) lectin II (CSA-II) was determined using a protein sequencer. After digestion of CSA-II with endoproteinase Lys-C or Asp-N, the resulting peptides were purified by reversed-phase high performance liquid chromatography (HPLC) and then subjected to sequence analysis. Comparison of the complete amino acid sequence of CSA-II with the sequences of other leguminous seed lectins revealed regions of extensive homology. The amino acid sequence of a putative carbohydrate-binding domain of CSA-II was found to be similar to those of several anti-H(O) leguminous lectins, especially to that of the L-fucose-binding Ulex europaeus lectin I (UEA-I).

  13. Management of NORM Residues

    International Nuclear Information System (INIS)

    2013-06-01

    The IAEA attaches great importance to the dissemination of information that can assist Member States in the development, implementation, maintenance and continuous improvement of systems, programmes and activities that support the nuclear fuel cycle and nuclear applications, and that address the legacy of past practices and accidents. However, radioactive residues are found not only in nuclear fuel cycle activities, but also in a range of other industrial activities, including: - Mining and milling of metalliferous and non-metallic ores; - Production of non-nuclear fuels, including coal, oil and gas; - Extraction and purification of water (e.g. in the generation of geothermal energy, as drinking and industrial process water; in paper and pulp manufacturing processes); - Production of industrial minerals, including phosphate, clay and building materials; - Use of radionuclides, such as thorium, for properties other than their radioactivity. Naturally occurring radioactive material (NORM) may lead to exposures at some stage of these processes and in the use or reuse of products, residues or wastes. Several IAEA publications address NORM issues with a special focus on some of the more relevant industrial operations. This publication attempts to provide guidance on managing residues arising from different NORM type industries, and on pertinent residue management strategies and technologies, to help Member States gain perspectives on the management of NORM residues

  14. Differential expression and localization of glycosidic residues in in vitro- and in vivo-matured cumulus-oocyte complexes in equine and porcine species.

    Science.gov (United States)

    Accogli, Gianluca; Douet, Cécile; Ambruosi, Barbara; Martino, Nicola Antonio; Uranio, Manuel Filioli; Deleuze, Stefan; Dell'Aquila, Maria Elena; Desantis, Salvatore; Goudet, Ghylène

    2014-12-01

    Glycoprotein oligosaccharides play major roles during reproduction, yet their function in gamete interactions is not fully elucidated. Identification and comparison of the glycan pattern in cumulus-oocyte complexes (COCs) from species with different efficiencies of in vitro spermatozoa penetration through the zona pellucida (ZP) could help clarify how oligosaccharides affect gamete interactions. We compared the expression and localization of 12 glycosidic residues in equine and porcine in vitro-matured (IVM) and preovulatory COCs by means of lectin histochemistry. The COCs glycan pattern differed between animals and COC source (IVM versus preovulatory). Among the 12 carbohydrate residues investigated, the IVM COCs from these two species shared: (a) sialo- and βN-acetylgalactosamine (GalNAc)-terminating glycans in the ZP; (b) sialylated and fucosylated glycans in cumulus cells; and (c) GalNAc and N-acetylglucosamine (GlcNAc) glycans in the ooplasm. Differences in the preovulatory COCs of the two species included: (a) sialoglycans and GlcNAc terminating glycans in the equine ZP versus terminal GalNAc and internal GlcNAc in the porcine ZP; (b) terminal galactosides in equine cumulus cells versus terminal GlcNAc and fucose in porcine cohorts; and (c) fucose in the mare ooplasm versus lactosamine and internal GlcNAc in porcine oocyte cytoplasm. Furthermore, equine and porcine cumulus cells and oocytes contributed differently to the synthesis of ZP glycoproteins. These results could be attributed to the different in vitro fertilization efficiencies between these two divergent, large-animal models. © 2014 Wiley Periodicals, Inc.

  15. Residual-stress measurements

    Energy Technology Data Exchange (ETDEWEB)

    Ezeilo, A N; Webster, G A [Imperial College, London (United Kingdom); Webster, P J [Salford Univ. (United Kingdom)

    1997-04-01

    Because neutrons can penetrate distances of up to 50 mm in most engineering materials, this makes them unique for establishing residual-stress distributions non-destructively. D1A is particularly suited for through-surface measurements as it does not suffer from instrumental surface aberrations commonly found on multidetector instruments, while D20 is best for fast internal-strain scanning. Two examples for residual-stress measurements in a shot-peened material, and in a weld are presented to demonstrate the attractive features of both instruments. (author).

  16. Old radioactive waste storage sites

    International Nuclear Information System (INIS)

    2008-01-01

    After a recall of the regulatory context for the management of old sites used for the storage of radioactive wastes with respect with their activity, the concerned products, the disposal or storage type, this document describes AREVA's involvement in the radioactive waste management process in France. Then, for the different kinds of sites (currently operated sites having radioactive waste storage, storage sites for uranium mineral processing residues), it indicates their location and name, their regulatory status and their control authority, the reference documents. It briefly presents the investigation on the long term impact of uranium mineral processing residues on health and environment, evokes some aspects of public information transparency, and presents the activities of an expertise group on old uranium mines. The examples of the sites of Bellezane (uranium mineral processing residues) and COMURHEX Malvesi (assessment of underground and surface water quality at the vicinity of this installation) are given in appendix

  17. Functional Differentiation of Antiporter-Like Polypeptides in Complex I; a Site-Directed Mutagenesis Study of Residues Conserved in MrpA and NuoL but Not in MrpD, NuoM, and NuoN.

    Directory of Open Access Journals (Sweden)

    Eva Sperling

    Full Text Available It has long been known that the three largest subunits in the membrane domain (NuoL, NuoM and NuoN of complex I are homologous to each other, as well as to two subunits (MrpA and MrpD from a Na+/H+ antiporter, Mrp. MrpA and NuoL are more similar to each other and the same is true for MrpD and NuoN. This suggests a functional differentiation which was proven experimentally in a deletion strain model system, where NuoL could restore the loss of MrpA, but not that of MrpD and vice versa. The simplest explanation for these observations was that the MrpA and MrpD proteins are not antiporters, but rather single subunit ion channels that together form an antiporter. In this work our focus was on a set of amino acid residues in helix VIII, which are only conserved in NuoL and MrpA (but not in any of the other antiporter-like subunits. and to compare their effect on the function of these two proteins. By combining complementation studies in B. subtilis and 23Na-NMR, response of mutants to high sodium levels were tested. All of the mutants were able to cope with high salt levels; however, all but one mutation (M258I/M225I showed differences in the efficiency of cell growth and sodium efflux. Our findings showed that, although very similar in sequence, NuoL and MrpA seem to differ on the functional level. Nonetheless the studied mutations gave rise to interesting phenotypes which are of interest in complex I research.

  18. Designing with residual materials

    NARCIS (Netherlands)

    Walhout, W.; Wever, R.; Blom, E.; Addink-Dölle, L.; Tempelman, E.

    2013-01-01

    Many entrepreneurial businesses have attempted to create value based on the residual material streams of third parties. Based on ‘waste’ materials they designed products, around which they built their company. Such activities have the potential to yield sustainable products. Many of such companies

  19. ABH antigens as recognition sites for the activation of red blood cell anion exchange by the lectin ulex europaeus agglutinin I.

    Science.gov (United States)

    Engelmann, B

    1993-11-01

    The blood group antigen H (blood group O) and fucose-specific lectin Ulex europaeus agglutinin I (UEA1) (10 micrograms/ml) was found to increase the rate constant of Cl- efflux into 100 mM Na+ oxalate media by about 40% in erythrocytes taken from antigen H donors. In 100 mM K+ oxalate, 150 mM Na+ pyruvate and in 150 mM Na+ acetate media the lectin elevated the rate constant of Cl- efflux by 20-50%. The acceleration of Cl- efflux by UEA1 was completely blocked by 10 microM 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid (DIDS) indicating that the effect of the lectin is mediated by the anion exchanger of human erythrocytes (band 3 protein). In antigen A1 erythrocytes no significant stimulation of anion exchange by UEA1 was seen. The activation of Cl- efflux was completely prevented by addition of 1 mM fucose to the medium. These results suggest that the effect of UEA1 is mediated through interaction with the fucose residues of H antigens. Increasing extracellular Ca++ from 0.5 to 5 mM in Na+ pyruvate or Na+ acetate media slightly reduced the acceleration of anion exchange by the lectin. On the other hand, replacing part of extracellular chloride by bicarbonate did not considerably alter the (previously reported) stimulatory effect of UEA1 on red blood cell Ca++ uptake. This suggests that the acceleration of anion exchange and of Ca++ uptake by UEA1, respectively, are mediated by different mechanisms. It is concluded that UEA1 activates anion exchange of human erythrocytes most probably by a direct interaction with H antigens present on extracellular domains of the band 3 protein.

  20. Measuring site occupancy

    DEFF Research Database (Denmark)

    Rogowska-Wrzesinska, Adelina; Wojdyla, Katarzyna; Williamson, James

    2014-01-01

    occupancy of the modification site. We show that, on one hand, heavily modified cysteines are not necessarily involved in the response to oxidative stress. On the other hand residues with low modification level can be dramatically affected by mild oxidative imbalance. We make use of high resolution mass...... peptides corresponding to 90 proteins. Only 6 modified peptides changed significantly under mild oxidative stress. Quantitative information allowed us to determine relative modification site occupancy of each identified modified residue and pin point heavily modified ones. The method proved to be precise...... and sensitive enough to detect and quantify endogenous levels of oxidative stress on proteome-wide scale and brings a new perspective on the role of the modification site occupancy in cellular redox response....

  1. Evaluation of residue-residue contact predictions in CASP9

    KAUST Repository

    Monastyrskyy, Bohdan; Fidelis, Krzysztof; Tramontano, Anna; Kryshtafovych, Andriy

    2011-01-01

    This work presents the results of the assessment of the intramolecular residue-residue contact predictions submitted to CASP9. The methodology for the assessment does not differ from that used in previous CASPs, with two basic evaluation measures

  2. Sharing Residual Liability

    DEFF Research Database (Denmark)

    Carbonara, Emanuela; Guerra, Alice; Parisi, Francesco

    2016-01-01

    Economic models of tort law evaluate the efficiency of liability rules in terms of care and activity levels. A liability regime is optimal when it creates incentives to maximize the value of risky activities net of accident and precaution costs. The allocation of primary and residual liability...... for policy makers and courts in awarding damages in a large number of real-world accident cases....

  3. Detecting coevolving amino acid sites using Bayesian mutational mapping

    DEFF Research Database (Denmark)

    Dimmic, Matthew W.; Hubisz, Melissa J.; Bustamente, Carlos D.

    2005-01-01

    Motivation: The evolution of protein sequences is constrained by complex interactions between amino acid residues. Because harmful substitutions may be compensated for by other substitutions at neighboring sites, residues can coevolve. We describe a Bayesian phylogenetic approach to the detection...

  4. Manual for implementing residual radioactivity guidelines

    International Nuclear Information System (INIS)

    Gilbert, T.L.; Eckerman, K.F.; Hansen, W.R.; Healy, J.W.; Kennedy, W.E.; Napier, B.A.; Solday, J.K.

    1986-01-01

    The US Department of Energy (DOE) has recently issued guidelines for residual radioactivity at Formerly Utilized Sites Remedial Action Program (FUSRAP) and remote Surplus Facilities Management Program (SFMP) sites. A manual for implementing these guidelines has been prepared jointly by four DOE laboratories (ANL, LANL, ORNL, and PNL) and is being issued as a supplement to the guidelines. The manual presents procedures and tables for deriving site-specific guidelines for levels of residual radionuclide concentrations in soil that must not be exceeded if a site is to be released for unrestricted use. Guidance for implementing DOE ALARA policy for remedial actions is also included. The concentration factor method is used in the pathway analysis for deriving soil guidelines. The analysis has been structured in a manner that explicitly identifies all of the factors involved. Tables are provided for dose-conversion factors and pathway factors from which environmental transport factors for each radionuclide and pathway may be calculated. The scenarios used for deriving the environmental transport factors and dose conversion factors, and the manner in which the information provided in the manual is used to derive site-specific soil guidelines will be presented

  5. Machine for compacting solid residues

    International Nuclear Information System (INIS)

    Herzog, J.

    1981-11-01

    Machine for compacting solid residues, particularly bulky radioactive residues, constituted of a horizontally actuated punch and a fixed compression anvil, in which the residues are first compacted horizontally and then vertically. Its salient characteristic is that the punch and the compression anvil have embossments on the compression side and interpenetrating plates in the compression position [fr

  6. Quadratic residues and non-residues selected topics

    CERN Document Server

    Wright, Steve

    2016-01-01

    This book offers an account of the classical theory of quadratic residues and non-residues with the goal of using that theory as a lens through which to view the development of some of the fundamental methods employed in modern elementary, algebraic, and analytic number theory. The first three chapters present some basic facts and the history of quadratic residues and non-residues and discuss various proofs of the Law of Quadratic Reciprosity in depth, with an emphasis on the six proofs that Gauss published. The remaining seven chapters explore some interesting applications of the Law of Quadratic Reciprocity, prove some results concerning the distribution and arithmetic structure of quadratic residues and non-residues, provide a detailed proof of Dirichlet’s Class-Number Formula, and discuss the question of whether quadratic residues are randomly distributed. The text is a valuable resource for graduate and advanced undergraduate students as well as for mathematicians interested in number theory.

  7. Expression of Ulex europaeus agglutinin I lectin-binding sites in squamous cell carcinomas and their absence in basal cell carcinomas. Indicator of tumor type and differentiation.

    Science.gov (United States)

    Heng, M C; Fallon-Friedlander, S; Bennett, R

    1992-06-01

    Lectins bind tightly to carbohydrate moieties on cell surfaces. Alterations in lectin binding have been reported to accompany epidermal cell differentiation, marking alterations in membrane sugars during this process. The presence of UEA I (Ulex europaeus agglutinin I) L-fucose-specific lectin-binding sites has been used as a marker for terminally differentiated (committed) keratinocytes. In this article, we report the presence of UEA-I-binding sites on squamous keratinocytes of well-differentiated squamous cell carcinomas, with patchy loss of UEA I positivity on poorly differentiated cells of squamous cell carcinomas, suggesting a possible use for this technique in the rapid assessment of less differentiated areas within the squamous cell tumor. The absence of UEA-I-binding sites on basal cell carcinomas may be related to an inability of cells comprising this tumor to convert the L-D-pyranosyl moiety on basal cells to the L-fucose moiety, resulting in an inability of basal cell carcinoma cell to undergo terminal differentiation into a committed keratinocyte.

  8. Stress and Fatigue Life Modeling of Cannon Breech Closures Including Effects of Material Strength and Residual Stress

    National Research Council Canada - National Science Library

    Underwood, John

    2000-01-01

    ...; overload residual stress. Modeling of applied and residual stresses at the location of the fatigue failure site is performed by elastic-plastic finite element analysis using ABAQUS and by solid...

  9. Complete Deletion of the Fucose Operon in Haemophilus influenzae Is Associated with a Cluster in Multilocus Sequence Analysis-Based Phylogenetic Group II Related to Haemophilus haemolyticus: Implications for Identification and Typing.

    Science.gov (United States)

    de Gier, Camilla; Kirkham, Lea-Ann S; Nørskov-Lauritsen, Niels

    2015-12-01

    Nonhemolytic variants of Haemophilus haemolyticus are difficult to differentiate from Haemophilus influenzae despite a wide difference in pathogenic potential. A previous investigation characterized a challenging set of 60 clinical strains using multiple PCRs for marker genes and described strains that could not be unequivocally identified as either species. We have analyzed the same set of strains by multilocus sequence analysis (MLSA) and near-full-length 16S rRNA gene sequencing. MLSA unambiguously allocated all study strains to either of the two species, while identification by 16S rRNA sequence was inconclusive for three strains. Notably, the two methods yielded conflicting identifications for two strains. Most of the "fuzzy species" strains were identified as H. influenzae that had undergone complete deletion of the fucose operon. Such strains, which are untypeable by the H. influenzae multilocus sequence type (MLST) scheme, have sporadically been reported and predominantly belong to a single branch of H. influenzae MLSA phylogenetic group II. We also found evidence of interspecies recombination between H. influenzae and H. haemolyticus within the 16S rRNA genes. Establishing an accurate method for rapid and inexpensive identification of H. influenzae is important for disease surveillance and treatment. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  10. REPLACEMENT OF TRYPTOPHAN RESIDUES IN HALOALKANE DEHALOGENASE REDUCES HALIDE BINDING AND CATALYTIC ACTIVITY

    NARCIS (Netherlands)

    KENNES, C; PRIES, F; KROOSHOF, GH; BOKMA, E; Kingma, Jacob; JANSSEN, DB

    1995-01-01

    Haloalkane dehalogenase catalyzes the hydrolytic cleavage of carbon-halogen bonds in short-chain haloalkanes. Two tryptophan residues of the enzyme (Trp125 and Trp175) form a halide-binding site in the active-site cavity, and were proposed to play a role in catalysis. The function of these residues

  11. Endosulfan residues in fish from the MUDA rice agroecosystem

    International Nuclear Information System (INIS)

    Nashriyah Mat; Sabri Junoh; Nuriati Nurdin

    2002-01-01

    A study was carried out in the recycled and the non-recycled areas of the Muda rice agroecosystem to determine the residue level of endosulfan in rice field fish. Snakehead or haruan (Channa striata), climbing perch or puyu (Anabas testudineus), walking catfish or keli (Clariasspp.) and snakeskin gouramy or sepat siam (Trichogasterpectoralis) were sampled and the tissues analysed using Gas Liquid Chromatography. Endosulfan sulphate metabolite was a ubiquitous residue in all fish at levels ranging from 0.0065 μg/g to 0.9837 μg/g of tissue wet weight whilst α-isomer was detected at a much lower concentration followed by β-isomer. In Channa striata, endosulfan total residue (α+β+sulphate) was accumulated in fish tissues sampled at ACRBD4 (non-recycled) site followed by Blok 14 (recycled) site. Endosulfan residue concentration did not coincide with water recycling practice nor changes in EROD enzyme activity. (Author)

  12. Bioenergy from sisal residues

    Energy Technology Data Exchange (ETDEWEB)

    Jungersen, G. [Dansk Teknologisk Inst. (Denmark); Kivaisi, A.; Rubindamayugi, M. [Univ. of Dar es Salaam (Tanzania, United Republic of)

    1998-05-01

    The main objectives of this report are: To analyse the bioenergy potential of the Tanzanian agro-industries, with special emphasis on the Sisal industry, the largest producer of agro-industrial residues in Tanzania; and to upgrade the human capacity and research potential of the Applied Microbiology Unit at the University of Dar es Salaam, in order to ensure a scientific and technological support for future operation and implementation of biogas facilities and anaerobic water treatment systems. The experimental work on sisal residues contains the following issues: Optimal reactor set-up and performance; Pre-treatment methods for treatment of fibre fraction in order to increase the methane yield; Evaluation of the requirement for nutrient addition; Evaluation of the potential for bioethanol production from sisal bulbs. The processing of sisal leaves into dry fibres (decortication) has traditionally been done by the wet processing method, which consumes considerable quantities of water and produces large quantities of waste water. The Tanzania Sisal Authority (TSA) is now developing a dry decortication method, which consumes less water and produces a waste product with 12-15% TS, which is feasible for treatment in CSTR systems (Continously Stirred Tank Reactors). (EG)

  13. Remaining Sites Verification Package for the 128-B-3 Burn Pit Site, Waste Site Reclassification Form 2006-058

    Energy Technology Data Exchange (ETDEWEB)

    L. M. Dittmer

    2006-11-17

    The 128-B-3 waste site is a former burn and disposal site for the 100-B/C Area, located adjacent to the Columbia River. The 128-B-3 waste site has been remediated to meet the remedial action objectives specified in the Remaining Sites ROD. The results of verification sampling demonstrated that residual contaminant concentrations do not preclude any future uses and allow for unrestricted use of shallow zone soils. The results of sampling at upland areas of the site also showed that residual contaminant concentrations are protective of groundwater and the Columbia River.

  14. Distributional map of the terminal and sub-terminal sugar residues of the glycoconjugates in the prepubertal and postpubertal testis of a subject affected by complete androgen insensitivity syndrome (Morris's syndrome): lectin histochemical study.

    Science.gov (United States)

    Gheri, G; Vannelli, G B; Marini, M; Zappoli Thyrion, G D; Gheri, R G; Sgambati, E

    2004-01-01

    In the present research we have investigated the distribution of the sugar residues of the glycoconjugates in the prepubertal and postpubertal testes of a subject with Morris's syndrome (CAIS, Complete Androgen Insensitivity Syndrome). For this purpose a battery of six horseradish peroxidase-conjugated lectins was used (SBA, PNA, WGA, ConA, LTA and UEAI). We have obtained a complete distributional map of the terminal and sub-terminal oligosaccharides in the tunica albuginea, interstitial tissue, lamina propria of the seminiferous tubules, Leydig cells, Sertoli cells, spermatogonia, mastocytes and endothelial cells. Furthermore the present study has shown that a large amount of sugar residues were detectable in the prepubertal and postpubertal testes but that some differences exist with particular regard to the Sertoli cells. The Sertoli cells and the Leydig cells of the retained prepubertal testis of the patient affected by Morris's syndrome were characterized by the presence of alpha-L-fucose, which was absent in the retained prepubertal testis of the normal subjects. Comparing the results on the postpubertal testis with those obtained on the same aged testis of healthy subjects we have demonstrated that alpha-L-fucose in the Sertoli and Leydig cells and D-galactose-N-acetyl-D-galactosamine in the Leydig cells are a unique feature of the subject affected by Morris's syndrome. D-galactose (ss1,3)-N-acetyl-D-galactosamine and sialic acid, which are present in the Leydig cells of the normal testis were never observed in the same cells of the postpubertal testis of the CAIS patient.

  15. Immobilization of acid digestion residue

    International Nuclear Information System (INIS)

    Greenhalgh, W.O.; Allen, C.R.

    1983-01-01

    Acid digestion treatment of nuclear waste is similar to incineration processes and results in the bulk of the waste being reduced in volume and weight to some residual solids termed residue. The residue is composed of various dispersible solid materials and typically contains the resultant radioactivity from the waste. This report describes the immobilization of the residue in portland cement, borosilicate glass, and some other waste forms. Diagrams showing the cement and glass virtification parameters are included in the report as well as process steps and candidate waste product forms. Cement immobilization is simplest and probably least expensive; glass vitrification exhibits the best overall volume reduction ratio

  16. Relocation of radioactive residuals store: environment effects statement

    International Nuclear Information System (INIS)

    1984-11-01

    This Environment Effects Statement describes and assesses the likely environmental effects of the proposal to relocate the Health Commission's existing radioactive residuals store to a site within the established Dutson Downs waste disposal area, located 20 km south-east of Sale and 225 km east of Melbourne. The information presented demonstrates that the siting and construction of the proposed radioactive residuals store and the procedures to be adopted for the handling and storage of materials will not present an unacceptable risk to public health and safety, nor will it involve any significant adverse environmental effects

  17. Evaluation of residue-residue contact predictions in CASP9

    KAUST Repository

    Monastyrskyy, Bohdan

    2011-01-01

    This work presents the results of the assessment of the intramolecular residue-residue contact predictions submitted to CASP9. The methodology for the assessment does not differ from that used in previous CASPs, with two basic evaluation measures being the precision in recognizing contacts and the difference between the distribution of distances in the subset of predicted contact pairs versus all pairs of residues in the structure. The emphasis is placed on the prediction of long-range contacts (i.e., contacts between residues separated by at least 24 residues along sequence) in target proteins that cannot be easily modeled by homology. Although there is considerable activity in the field, the current analysis reports no discernable progress since CASP8.

  18. Multiple Site Damage in Flat Panel Testing

    National Research Council Canada - National Science Library

    Shrage, Daniel

    2000-01-01

    This report aimed to experimentally verify analytical models that predict the residual strength of representative aircraft structures, such as wide panels, that are subjected to Multiple Site Damage (MSD...

  19. Residual radioactive material guidelines: Methodology and applications

    International Nuclear Information System (INIS)

    Yu, C.; Yuan, Y.C.; Zielen, A.J.; Wallo, A. III.

    1989-01-01

    A methodology to calculate residual radioactive material guidelines was developed for the US Department of Energy (DOE). This methodology is coded in a menu-driven computer program, RESRAD, which can be run on IBM or IBM-compatible microcomputers. Seven pathways of exposure are considered: external radiation, inhalation, and ingestion of plant foods, meat, milk, aquatic foods, and water. The RESRAD code has been applied to several DOE sites to calculate soil cleanup guidelines. This experience has shown that the computer code is easy to use and very user-friendly. 3 refs., 8 figs

  20. Landfilling of waste incineration residues

    DEFF Research Database (Denmark)

    Christensen, Thomas Højlund; Astrup, Thomas; Cai, Zuansi

    2002-01-01

    Residues from waste incineration are bottom ashes and air-pollution-control (APC) residues including fly ashes. The leaching of heavy metals and salts from the ashes is substantial and a wide spectrum of leaching tests and corresponding criteria have been introduced to regulate the landfilling...

  1. Statistical inference on residual life

    CERN Document Server

    Jeong, Jong-Hyeon

    2014-01-01

    This is a monograph on the concept of residual life, which is an alternative summary measure of time-to-event data, or survival data. The mean residual life has been used for many years under the name of life expectancy, so it is a natural concept for summarizing survival or reliability data. It is also more interpretable than the popular hazard function, especially for communications between patients and physicians regarding the efficacy of a new drug in the medical field. This book reviews existing statistical methods to infer the residual life distribution. The review and comparison includes existing inference methods for mean and median, or quantile, residual life analysis through medical data examples. The concept of the residual life is also extended to competing risks analysis. The targeted audience includes biostatisticians, graduate students, and PhD (bio)statisticians. Knowledge in survival analysis at an introductory graduate level is advisable prior to reading this book.

  2. Residual Stresses and Other Properties of Teardrops

    Energy Technology Data Exchange (ETDEWEB)

    Stroud, Mary Ann [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Veirs, Douglas Kirk [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Berg, John M. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Hill, Mary Ann [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Rios, Daniel [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Duque, Juan [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2017-07-26

    The Department of Energy’s 3013 Standard for packaging plutonium-bearing materials for storage up to fifty years specifies a minimum of two individually welded, nested containers herein referred to as the 3013 outer and the 3013 inner.1 Stress corrosion cracking (SCC) is a potential failure mechanism for 3013 inner containers.2,3 The bagless transfer container (BTC), a 3013 inner container used by Hanford and Savanna River Site (SRS) made from 304L stainless steel (SS), poses the greatest concern for SCC.4,5 The Surveillance and Monitoring Program (SMP) use stressed metal samples known as teardrops as screening tools in SCC studies to evaluate factors that could result in cracks in the 3013 containers.6,7 This report provides background information on the teardrops used in the Los Alamos National Laboratory (LANL) SMP studies including method of construction, composition and variability. In addition, the report discusses measurements of residual stresses in teardrops and compares the results with residual stresses in BTCs reported previously.4 Factors affecting residual stresses, including teardrop dimensions and surface finish, are also discussed.

  3. Identification of NAD interacting residues in proteins

    Directory of Open Access Journals (Sweden)

    Raghava Gajendra PS

    2010-03-01

    Full Text Available Abstract Background Small molecular cofactors or ligands play a crucial role in the proper functioning of cells. Accurate annotation of their target proteins and binding sites is required for the complete understanding of reaction mechanisms. Nicotinamide adenine dinucleotide (NAD+ or NAD is one of the most commonly used organic cofactors in living cells, which plays a critical role in cellular metabolism, storage and regulatory processes. In the past, several NAD binding proteins (NADBP have been reported in the literature, which are responsible for a wide-range of activities in the cell. Attempts have been made to derive a rule for the binding of NAD+ to its target proteins. However, so far an efficient model could not be derived due to the time consuming process of structure determination, and limitations of similarity based approaches. Thus a sequence and non-similarity based method is needed to characterize the NAD binding sites to help in the annotation. In this study attempts have been made to predict NAD binding proteins and their interacting residues (NIRs from amino acid sequence using bioinformatics tools. Results We extracted 1556 proteins chains from 555 NAD binding proteins whose structure is available in Protein Data Bank. Then we removed all redundant protein chains and finally obtained 195 non-redundant NAD binding protein chains, where no two chains have more than 40% sequence identity. In this study all models were developed and evaluated using five-fold cross validation technique on the above dataset of 195 NAD binding proteins. While certain type of residues are preferred (e.g. Gly, Tyr, Thr, His in NAD interaction, residues like Ala, Glu, Leu, Lys are not preferred. A support vector machine (SVM based method has been developed using various window lengths of amino acid sequence for predicting NAD interacting residues and obtained maximum Matthew's correlation coefficient (MCC 0.47 with accuracy 74.13% at window length 17

  4. Residual stress by repair welds

    International Nuclear Information System (INIS)

    Mochizuki, Masahito; Toyoda, Masao

    2003-01-01

    Residual stress by repair welds is computed using the thermal elastic-plastic analysis with phase-transformation effect. Coupling phenomena of temperature, microstructure, and stress-strain fields are simulated in the finite-element analysis. Weld bond of a plate butt-welded joint is gouged and then deposited by weld metal in repair process. Heat source is synchronously moved with the deposition of the finite-element as the weld deposition. Microstructure is considered by using CCT diagram and the transformation behavior in the repair weld is also simulated. The effects of initial stress, heat input, and weld length on residual stress distribution are studied from the organic results of numerical analysis. Initial residual stress before repair weld has no influence on the residual stress after repair treatment near weld metal, because the initial stress near weld metal releases due to high temperature of repair weld and then stress by repair weld regenerates. Heat input has an effect for residual stress distribution, for not its magnitude but distribution zone. Weld length should be considered reducing the magnitude of residual stress in the edge of weld bead; short bead induces high tensile residual stress. (author)

  5. RESIDUAL RISK ASSESSMENT: ETHYLENE OXIDE ...

    Science.gov (United States)

    This document describes the residual risk assessment for the Ethylene Oxide Commercial Sterilization source category. For stationary sources, section 112 (f) of the Clean Air Act requires EPA to assess risks to human health and the environment following implementation of technology-based control standards. If these technology-based control standards do not provide an ample margin of safety, then EPA is required to promulgate addtional standards. This document describes the methodology and results of the residual risk assessment performed for the Ethylene Oxide Commercial Sterilization source category. The results of this analyiss will assist EPA in determining whether a residual risk rule for this source category is appropriate.

  6. Predicting logging residues: an interim equation for Appalachian oak sawtimber

    Science.gov (United States)

    A. Jeff Martin

    1975-01-01

    An equation, using dbh, dbh², bole length, and sawlog height to predict the cubic-foot volume of logging residue per tree, was developed from data collected on 36 mixed oaks in southwestern Virginia. The equation produced reliable results for small sawtimber trees, but additional research is needed for other species, sites, and utilization practices.

  7. Quantification and characterization of cotton crop biomass residue

    Science.gov (United States)

    Cotton crop residual biomass remaining in the field after mechanical seed cotton harvest is not typically harvested and utilized off-site thereby generating additional revenue for producers. Recently, interest has increased in utilizing biomass materials as feedstock for the production of fuel and ...

  8. CONSTRUCTION OF A FUNCTIONAL LACTOSE PERMEASE DEVOID OF CYSTEINE RESIDUES

    NARCIS (Netherlands)

    VANIWAARDEN, PR; PASTORE, JC; KONINGS, WN; KABACK, HR

    1991-01-01

    By use of oligonucleotide-directed, site-specific mutagenesis, a lactose (lac) permease molecule was constructed in which all eight cysteinyl residues were simultaneously mutagenized (C-less permease). Cys154 was replaced with valine, and Cys117, -148, -176, -234, -333, -353, and -355 were replaced

  9. Disposal of residue from uranium ore processing in France

    International Nuclear Information System (INIS)

    Crochon, Ph.

    2011-01-01

    Between 1949 and 2001, French mines produced 76, 000 metric tons of uranium and 50 million metric tons of ore, processing residues are stored at 17 sites (in ponds enclosed by dykes or in former open-cast mines) subject to ICPE (classified facility for environment protection) regulation. These disposal sites cover surface areas of between one and several tens of hectares and several thousands to several millions of metric tons of waste are stored at them. When uranium mining stopped in France, these sites were redeveloped, with caps placed over the residue to provide mechanical and radiological protection. All these sites are still monitored by AREVA. In the last fifteen years, these sites have been the subject of a number of studies, especially regarding the long-term evolution and impact of the residue. These studies are now being pursued within the framework of the national plan for the management of nuclear materials and waste (PNGMDR). A regulatory and institutional framework regarding long-term management of these disposal sites needs to be defined. (author)

  10. Investigating the properties of residues. Characterization of pellets from fermentation residues; Den Eigenschaften der Reststoffe auf der Spur. Untersuchung widmet sich der Charakterisierung von Pellets aus Gaerresten

    Energy Technology Data Exchange (ETDEWEB)

    Kratzeisen, Martin; Mueller, Joachim [Hohenheim Univ., Stuttgart (Germany). Inst. fuer Agrartechnik; Starcevic, Nikica [Hohenheim Univ., Stuttgart (Germany). Inst. fuer Agrartechnik; Strabag Umweltanlagen GmbH, Muenchen (Germany). Projekt Produktentwicklung/Schlammbehandlung

    2009-09-15

    Fermentation residues are by-products of the biogas process. Farmers use them as fertilizers, but as the size of biogas plants grows, so does the residues volume. It is now too much for local use, and transport to other sites is expensive. Fuel pellets production may be an alternative. Pellets from fermentation residues are not accepted as yet because too little is known about their characteristics. The contribution describes an investigation that intends to identify the fuel characteristics of pellets from fermentation residues. (orig.)

  11. Evaluation and optimization of nutritional and environmental impact of biogas residues

    International Nuclear Information System (INIS)

    Lichti, Fabian Heribert

    2013-01-01

    On the basis of the dynamic growth of biogas plants in Germany the fertilization with biogas residues has obtained an important role for recirculation of plant nutrients, particularly with regard to nitrogen. In this work the effect of N nutrition with biogas residues was assessed in a 3-year on-field trial conducted at four sites throughout Bavaria. The fertilizing effects were tested by varying rate and time of biogas residues application, using different application techniques and the addition of nitrification inhibitors on several crops. The biogas residues achieved mineral fertilizer equivalents of 30 - 45 %. Overall, the untreated biogas residues showed a slightly increased N efficiency compared to cattle manure, whereas particularly site-dependent differences resulted in large differences in N efficiency of biogas residues.

  12. Identification of Key Residues for Enzymatic Carboxylate Reduction

    Directory of Open Access Journals (Sweden)

    Holly Stolterfoht

    2018-02-01

    Full Text Available Carboxylate reductases (CARs, E.C. 1.2.1.30 generate aldehydes from their corresponding carboxylic acid with high selectivity. Little is known about the structure of CARs and their catalytically important amino acid residues. The identification of key residues for carboxylate reduction provides a starting point to gain deeper understanding of enzymatic carboxylate reduction. A multiple sequence alignment of CARs with confirmed activity recently identified in our lab and from the literature revealed a fingerprint of conserved amino acids. We studied the function of conserved residues by multiple sequence alignments and mutational replacements of these residues. In this study, single-site alanine variants of Neurospora crassa CAR were investigated to determine the contribution of conserved residues to the function, expressability or stability of the enzyme. The effect of amino acid replacements was investigated by analyzing enzymatic activity of the variants in vivo and in vitro. Supported by molecular modeling, we interpreted that five of these residues are essential for catalytic activity, or substrate and co-substrate binding. We identified amino acid residues having significant impact on CAR activity. Replacement of His 237, Glu 433, Ser 595, Tyr 844, and Lys 848 by Ala abolish CAR activity, indicating their key role in acid reduction. These results may assist in the functional annotation of CAR coding genes in genomic databases. While some other conserved residues decreased activity or had no significant impact, four residues increased the specific activity of NcCAR variants when replaced by alanine. Finally, we showed that NcCAR wild-type and mutants efficiently reduce aliphatic acids.

  13. Criteria for the restoration of mining residues in Germany

    International Nuclear Information System (INIS)

    Kraus, W.; Ettenhuber, E.; Gehrcke, K.; Przyborowski, S.

    2000-01-01

    Residues from uranium mines and mills and from the mining of silver, tin, cobalt, nickel and other ores, as well as of coal mineralized with uranium, are situated in densely populated regions of Germany. Social and political pressure required an urgent investigation and evaluation of these residues in order to identify relevant residues which could not be disregarded from the radiation protection point of view. There were two categories of residues. First, for huge former uranium mining and milling sites, the original owner could be made liable for restoration. A large Federal rehabilitation programme for the Wismut sites was started immediately after the political change in the former East Germany in 1990 and was based on radiological as well as on social and economic concerns. Secondly, for a large number of smaller residues, sometimes dating back to the middle ages, an evaluation of their radiological relevance was necessary before decisions could be taken on the justification of a restoration. This was the objective of a Federal programme on registration, investigation and evaluation of mining residues. Up to now only minor remedial activities have been carried out in cases where an urgent need had been detected. Criteria developed by the German Commission on radiological protection (SSK) were applied for the evaluation of the residues. The primary criterion for the justification of a restoration was an annual individual effective dose of 1 mSv for all exposure pathways except for the inhalation of radon. For inhalation of radon, the primary criterion for justification was a long term average outdoor radon concentration of 50 Bq/m 3 caused by the residues. Both levels were taken in addition to the natural background radiation level at a given site. These criteria were based on the upper end of the 'normal' range of naturally occurring exposure or concentration levels. SSK established reference levels in measurable quantities (activity concentration in soil

  14. Nitrogen availability of biogas residues

    Energy Technology Data Exchange (ETDEWEB)

    El-Sayed Fouda, Sara

    2011-09-07

    The objectives of this study were to characterize biogas residues either unseparated or separated into a liquid and a solid phase from the fermentation of different substrates with respect to their N and C content. In addition, short and long term effects of the application of these biogas residues on the N availability and N utilization by ryegrass was investigated. It is concluded that unseparated or liquid separated biogas residues provide N at least corresponding to their ammonium content and that after the first fertilizer application the C{sub org}:N{sub org} ratio of the biogas residues was a crucial factor for the N availability. After long term application, the organic N accumulated in the soil leads to an increased release of N.

  15. Site Features

    Data.gov (United States)

    U.S. Environmental Protection Agency — This dataset consists of various site features from multiple Superfund sites in U.S. EPA Region 8. These data were acquired from multiple sources at different times...

  16. Vesícula residual

    Directory of Open Access Journals (Sweden)

    Júlio C. U. Coelho

    Full Text Available Our objective is to report three patients with recurrent severe upper abdominal pain secondary to residual gallbladder. All patients had been subjected to cholecystectomy from 1 to 20 years before. The diagnosis was established after several episodes of severe upper abdominal pain by imaging exams: ultrasonography, tomography, or endoscopic retrograde cholangiography. Removal of the residual gallbladder led to complete resolution of symptoms. Partial removal of the gallbladder is a very rare cause of postcholecystectomy symptoms.

  17. Residual number processing in dyscalculia ?

    OpenAIRE

    Cappelletti, Marinella; Price, Cathy J.

    2013-01-01

    Developmental dyscalculia – a congenital learning disability in understanding numerical concepts – is typically associated with parietal lobe abnormality. However, people with dyscalculia often retain some residual numerical abilities, reported in studies that otherwise focused on abnormalities in the dyscalculic brain. Here we took a different perspective by focusing on brain regions that support residual number processing in dyscalculia. All participants accurately performed semantic and ca...

  18. Americium recovery from reduction residues

    Science.gov (United States)

    Conner, W.V.; Proctor, S.G.

    1973-12-25

    A process for separation and recovery of americium values from container or bomb'' reduction residues comprising dissolving the residues in a suitable acid, adjusting the hydrogen ion concentration to a desired level by adding a base, precipitating the americium as americium oxalate by adding oxalic acid, digesting the solution, separating the precipitate, and thereafter calcining the americium oxalate precipitate to form americium oxide. (Official Gazette)

  19. Site decontamination

    International Nuclear Information System (INIS)

    Bicker, A.E.

    1981-01-01

    Among the several DOE sites that have been radiologically decontaminated under the auspices of the Nevada Operations Office are three whose physical characteristics are unique. These are the Tatum Dome Test Site (TDTS) near Hattiesburg, Mississippi; a location of mountainous terrain (Pahute Mesa) on the Nevada Test Site; and the GNOME site near Carlsbad, New Mexico. In each case the contamination, the terrain, and the climate conditions were different. This presentation includes a brief description of each site, the methods used to perform radiological surveys, the logistics required to support the decontamination (including health physics and sample analysis), and the specific techniques used to reduce or remove the contamination

  20. Evaluation of residue-residue contact prediction in CASP10

    KAUST Repository

    Monastyrskyy, Bohdan

    2013-08-31

    We present the results of the assessment of the intramolecular residue-residue contact predictions from 26 prediction groups participating in the 10th round of the CASP experiment. The most recently developed direct coupling analysis methods did not take part in the experiment likely because they require a very deep sequence alignment not available for any of the 114 CASP10 targets. The performance of contact prediction methods was evaluated with the measures used in previous CASPs (i.e., prediction accuracy and the difference between the distribution of the predicted contacts and that of all pairs of residues in the target protein), as well as new measures, such as the Matthews correlation coefficient, the area under the precision-recall curve and the ranks of the first correctly and incorrectly predicted contact. We also evaluated the ability to detect interdomain contacts and tested whether the difficulty of predicting contacts depends upon the protein length and the depth of the family sequence alignment. The analyses were carried out on the target domains for which structural homologs did not exist or were difficult to identify. The evaluation was performed for all types of contacts (short, medium, and long-range), with emphasis placed on long-range contacts, i.e. those involving residues separated by at least 24 residues along the sequence. The assessment suggests that the best CASP10 contact prediction methods perform at approximately the same level, and comparably to those participating in CASP9.

  1. Characterizing the glycocalyx of poultry spermatozoa: I. Identification and distribution of carbohydrate residues using flow cytometry and epifluorescence microscopy.

    Science.gov (United States)

    Peláez, Jesús; Long, Julie A

    2007-01-01

    The aim of the present work was to use a battery of lectins to 1) delineate the carbohydrate content of sperm glycocalyx in the turkey and chicken using flow cytometry analysis, and 2) evaluate the distribution of existing sugars over the sperm plasma membrane surface with epifluorescent microscopy. Carbohydrate groups (corresponding lectins) that were investigated included galactose (GS-I, Jacalin, RCA-I, PNA), glucose and/or mannose (Con A, PSA, GNA), N-acetyl-glucosamine (GS-II, s-WGA, STA), N-acetyl-galactosamine (SBA, WFA), fucose (Lotus, UEA-I), sialic acid (LFA, LPA), and N-acetyl-lactosamine (ECA). Spermatozoa were assessed before and after treatment with neuraminidase to remove sialic acid. Mean fluorescence intensity (MnFI) was used as indicator of lectin binding for flow cytometry analysis. Nontreated spermatozoa from both species showed high MnFI when incubated with RCA-I, Con A, LFA, and LPA, as did chicken spermatozoa incubated with s-WGA. Neuraminidase treatment increased the MnFI for most lectins except LFA and LPA, as expected. Differences in MnFI between species included higher values for s-WGA and ECA in chicken spermatozoa and for WFA in turkey spermatozoa. Microscopy revealed segregation of some sugar residues into membrane-specific domains; however, the 2 staining techniques (cell suspension vs fixed preparation) differed in identifying lectin binding patterns, with fixed preparations yielding a high degree of nonspecific binding. We conclude that 1) the glycocalyx of turkey and chicken spermatozoa contains a diversity of carbohydrate groups, 2) these residues are extensively masked by sialic acid, 3) the glycocalyx composition is species-specific, and 4) some glycoconjugates appear to be segregated into membrane-specific domains. Characterization of the poultry sperm glycocalyx is the first step in identifying the physiological impact of semen storage on sperm function.

  2. Remaining Sites Verification Package for 100-F-38 Stained Soil Site. Attachment to Waste Site Reclassification Form 2004-093

    International Nuclear Information System (INIS)

    Carlson, R.A.

    2006-01-01

    The 100-F-38 Stained Soil site was an area of yellow stained soil that was discovered while excavating a trench for the placement of electrical conduit. The 100-F-38 Stained Soil site meets the remedial action objectives specified in the Remaining Sites ROD. The results of verification sampling show demonstrate that residual contaminant concentrations support future unrestricted land uses that can be represented by a rural-residential scenario. The results also show that residual contaminant concentrations do not preclude any future uses and allow for unrestricted use of shallow zone soils and the contaminant concentrations remaining in the soil are protective of groundwater and the Columbia River

  3. Productivity and cost analysis of a mobile pyrolysis system deployed to convert mill residues into biochar

    Science.gov (United States)

    Woodam Chung; Dongyeob Kim; Nathaniel Anderson

    2012-01-01

    Forest and mill residues are a promising source of biomass feedstock for the production of bioenergy, biofuels and bioproducts. However, high costs of transportation and handling of feedstock often make utilization of forest residues, such as logging slash, financially unviable. As a result, these materials are often considered waste and left on site to decompose or...

  4. Diversity in crop residue management across an intensification gradient in southern Africa

    NARCIS (Netherlands)

    Rusinamhodzi, Leonard; Corbeels, Marc; Giller, Ken E.

    2016-01-01

    Crop residues are important for livestock feed and nutrient cycling among many other functions on smallholder farming systems of sub-Saharan Africa. The objective of this study was to assess differences in resource endowment, crop productivity and crop residue management in selected sites in

  5. Site organization and site arrangement

    International Nuclear Information System (INIS)

    Boissonnet, B.; Macqueron, J.F.

    1976-01-01

    The present paper deals with criteria for the choice of a production unit or power plant site, the organization and development of a site in terms of its particular characteristics and takes into account personnel considerations in site organizations as well as the problem of integrating the architecture into the environment. (RW) [de

  6. Measures for radiation prevention and remediation of islightly radioactive contaminated sites by phytoremediation and subsequent utilization of the loaded plant residues (PHYTOREST). Final report; Massnahmen zur Strahlenschutzvorsorge radioaktiv belasteter Grossflaechen durch Sanierung mittels Phytoremediation und anschliessende Verwertung der belasteten Pflanzenreststoffe (PHYTOREST). Abschlussbericht

    Energy Technology Data Exchange (ETDEWEB)

    Willscher, Sabine; Werner, Peter [TU Dresden, Pirna (Germany). Inst. fuer Abfallwirtschaft und Altlasten; Jablonski, Lukasz; Wittig, Juliane

    2013-12-30

    contaminated geosubstrates now can be carried out within the radiation protection regulations. Hence, the project provides a substantial contribution to the radiation protection of HM/R contaminated soils. Within the research project, ways for the utilization of HM/R- contaminated plant residuals were highlighted; this gives a substantial contribution for minimization of wastes, the winning of sustainable bioenergy and the recycling of materials. Here, different ways of solutions were investigated. The research project was carried out within the scientific funding program ''Closedown and decommissioning of nuclear facilities''. The results of the project will contribute to the development of a biologically benign, sustainable technique for the remediation of large contaminated areas that originate mostly from the legacy of the former U mining. As a general result of this comprehensive research project, a phytostabilization/ phytoextraction of such SM/R contaminated sites is feasible with a protection of ground water, and the plant crop from phytoremediation of the HM/R contaminated field site can be utilized for the winning of bioenergy (gaseous/ liquid products or thermal utilization). The beneficial combination of phytoremediation and subsequent utilization of the biomass can be further developed to an innovative and sustainable remediation technology with national and international application potential.

  7. LigandRFs: random forest ensemble to identify ligand-binding residues from sequence information alone

    KAUST Repository

    Chen, Peng; Huang, Jianhua Z; Gao, Xin

    2014-01-01

    Protein-ligand binding is important for some proteins to perform their functions. Protein-ligand binding sites are the residues of proteins that physically bind to ligands. Despite of the recent advances in computational prediction

  8. Residual stresses around Vickers indents

    International Nuclear Information System (INIS)

    Pajares, A.; Guiberteau, F.; Steinbrech, R.W.

    1995-01-01

    The residual stresses generated by Vickers indentation in brittle materials and their changes due to annealing and surface removal were studied in 4 mol% yttria partially stabilized zirconia (4Y-PSZ). Three experimental methods to gain information about the residual stress field were applied: (i) crack profile measurements based on serial sectioning, (ii) controlled crack propagation in post indentation bending tests and (iii) double indentation tests with smaller secondary indents located around a larger primary impression. Three zones of different residual stress behavior are deduced from the experiments. Beneath the impression a crack free spherical zone of high hydrostatic stresses exists. This core zone is followed by a transition regime where indentation cracks develop but still experience hydrostatic stresses. Finally, in an outward third zone, the crack contour is entirely governed by the tensile residual stress intensity (elastically deformed region). Annealing and surface removal reduce this crack driving stress intensity. The specific changes of the residual stresses due to the post indentation treatments are described and discussed in detail for the three zones

  9. Site operations

    International Nuclear Information System (INIS)

    House, W.B.; Ebenhack, D.G.

    1989-01-01

    This chapter is a discussion of the management and operations practices used at the Barnwell Waste Management Facility in Barnwell, SC. The following topics are discussed: (1) Waste receiving and inspection, including manifest and certificates of compliance, radiological surveys, disposition of nonconforming items, and decontamination and disposition of secondary waste streams; (2) Waste disposal, including Title 10 CFR 61 requirements, disposal area evaluations, shipment offloading, container emplacement, and radiation protection; (3) Trench closure, including trench backfilling, trench capping, and permanent markers; (4) Site maintenance and stabilization, including trench maintenance, surface water management, and site closure activities; (5) Site monitoring programs, including operational monitoring, and environmental monitoring program; (6) Personnel training and qualifications, including basic training program, safety training program, special skills training, and physical qualifications; (7) Records management, including waste records, personnel training records, personnel dosimetry records, site monitoring records, trench qualification and construction records, and site drawings and stabilization records; (8) Site security; (9) Emergency response plans; and (10) Quality assurance

  10. Hazardous properties of paint residues from the furniture industry.

    Science.gov (United States)

    Vaajasaari, Kati; Kulovaara, Maaret; Joutti, Anneli; Schultz, Eija; Soljamo, Kari

    2004-01-30

    The objective of this study was to screen nine excess paint residues for environmental hazard and to evaluate their disposability in a non-hazardous or hazardous-waste landfill. These residues were produced in the process of spray-painting furniture. Residues were classified according to their leaching and ecotoxicological properties. Leaching properties were determined with the European standard SFS-EN 12457-2 leaching-test. The toxicity of the leaching-test eluates was measured with plant-, bacteria- and enzyme-inhibition bioassays. Total organic carbon, formaldehyde and solvent concentrations in the solid wastes and in the leaching-test eluates were analysed. It seemed likely that leached formaldehyde caused very high acute toxicity in leaching-test eluates of the dry-booth residues. This hypothesis was based on the fact that the formaldehyde concentrations in the leaching-test eluates of the dry-booth residues were 62-75 times higher than the EC50 value reported in the literature for formaldehyde. The results of the water-curtain booth residues showed that the samples with the highest TOC and aromatic solvent concentrations were also the most toxic. The studied excess paint residues were complex organic mixtures and contained large amounts of compounds not identifiable from chemical data. Therefore, the evaluation of the hazard based solely on available chemical data is unlikely to be sufficient, as evidenced by our study. Our results show that harmful compounds remain in the solid waste and the toxicity results of their leaching-test eluates show that toxicity may leach from residues in contact with water at landfill sites. They also confirm the benefit of combining chemical and ecotoxicological assays in assessing the potential environmental hazard of complex organic mixtures found in wastes. Copyright 2003 Elsevier B.V.

  11. Actinide recovery from pyrochemical residues

    International Nuclear Information System (INIS)

    Avens, L.R.; Clifton, D.G.; Vigil, A.R.

    1984-01-01

    A new process for recovery of plutonium and americium from pyrochemical waste has been demonstrated. It is based on chloride solution anion exchange at low acidity, which eliminates corrosive HCl fumes. Developmental experiments of the process flowsheet concentrated on molten salt extraction (MSE) residues and gave >95% plutonium and >90% americium recovery. The recovered plutonium contained 6 = from high chloride-low acid solution. Americium and other metals are washed from the ion exchange column with 1N HNO 3 -4.8M NaCl. The plutonium is recovered, after elution, via hydroxide precipitation, while the americium is recovered via NaHCO 3 precipitation. All filtrates from the process are discardable as low-level contaminated waste. Production-scale experiments are now in progress for MSE residues. Flow sheets for actinide recovery from electrorefining and direct oxide reduction residues are presented and discussed

  12. Actinide recovery from pyrochemical residues

    International Nuclear Information System (INIS)

    Avens, L.R.; Clifton, D.G.; Vigil, A.R.

    1985-05-01

    We demonstrated a new process for recovering plutonium and americium from pyrochemical waste. The method is based on chloride solution anion exchange at low acidity, or acidity that eliminates corrosive HCl fumes. Developmental experiments of the process flow chart concentrated on molten salt extraction (MSE) residues and gave >95% plutonium and >90% americium recovery. The recovered plutonium contained 6 2- from high-chloride low-acid solution. Americium and other metals are washed from the ion exchange column with lN HNO 3 -4.8M NaCl. After elution, plutonium is recovered by hydroxide precipitation, and americium is recovered by NaHCO 3 precipitation. All filtrates from the process can be discardable as low-level contaminated waste. Production-scale experiments are in progress for MSE residues. Flow charts for actinide recovery from electro-refining and direct oxide reduction residues are presented and discussed

  13. Coking of residue hydroprocessing catalysts

    Energy Technology Data Exchange (ETDEWEB)

    Gray, M.R.; Zhao, Y.X. [Alberta Univ., Edmonton, AB (Canada). Dept. of Chemical Engineering; McKnight, C.A. [Syncrude Canada Ltd., Edmonton, AB (Canada); Komar, D.A.; Carruthers, J.D. [Cytec Industries Inc., Stamford, CT (United States)

    1997-11-01

    One of the major causes of deactivation of Ni/Mo and Co/Mo sulfide catalysts for hydroprocessing of heavy petroleum and bitumen fractions is coke deposition. The composition and amount of coke deposited on residue hydroprocessing catalysts depends on the composition of the liquid phase of the reactor. In the Athabasca bitumen, the high molecular weight components encourage coke deposition at temperatures of 430 to 440 degrees C and at pressures of 10 to 20 MPa hydrogen pressure. A study was conducted to determine which components in the heavy residual oil fraction were responsible for coking of catalysts. Seven samples of Athabasca vacuum residue were prepared by supercritical fluid extraction with pentane before being placed in the reactor. Carbon content and hydrodesulfurization activity was measured. It was concluded that the deposition of coke depended on the presence of asphaltenes and not on other compositional variables such as content of nitrogen, aromatic carbon or vanadium.

  14. Text mining improves prediction of protein functional sites.

    Directory of Open Access Journals (Sweden)

    Karin M Verspoor

    Full Text Available We present an approach that integrates protein structure analysis and text mining for protein functional site prediction, called LEAP-FS (Literature Enhanced Automated Prediction of Functional Sites. The structure analysis was carried out using Dynamics Perturbation Analysis (DPA, which predicts functional sites at control points where interactions greatly perturb protein vibrations. The text mining extracts mentions of residues in the literature, and predicts that residues mentioned are functionally important. We assessed the significance of each of these methods by analyzing their performance in finding known functional sites (specifically, small-molecule binding sites and catalytic sites in about 100,000 publicly available protein structures. The DPA predictions recapitulated many of the functional site annotations and preferentially recovered binding sites annotated as biologically relevant vs. those annotated as potentially spurious. The text-based predictions were also substantially supported by the functional site annotations: compared to other residues, residues mentioned in text were roughly six times more likely to be found in a functional site. The overlap of predictions with annotations improved when the text-based and structure-based methods agreed. Our analysis also yielded new high-quality predictions of many functional site residues that were not catalogued in the curated data sources we inspected. We conclude that both DPA and text mining independently provide valuable high-throughput protein functional site predictions, and that integrating the two methods using LEAP-FS further improves the quality of these predictions.

  15. Text Mining Improves Prediction of Protein Functional Sites

    Science.gov (United States)

    Cohn, Judith D.; Ravikumar, Komandur E.

    2012-01-01

    We present an approach that integrates protein structure analysis and text mining for protein functional site prediction, called LEAP-FS (Literature Enhanced Automated Prediction of Functional Sites). The structure analysis was carried out using Dynamics Perturbation Analysis (DPA), which predicts functional sites at control points where interactions greatly perturb protein vibrations. The text mining extracts mentions of residues in the literature, and predicts that residues mentioned are functionally important. We assessed the significance of each of these methods by analyzing their performance in finding known functional sites (specifically, small-molecule binding sites and catalytic sites) in about 100,000 publicly available protein structures. The DPA predictions recapitulated many of the functional site annotations and preferentially recovered binding sites annotated as biologically relevant vs. those annotated as potentially spurious. The text-based predictions were also substantially supported by the functional site annotations: compared to other residues, residues mentioned in text were roughly six times more likely to be found in a functional site. The overlap of predictions with annotations improved when the text-based and structure-based methods agreed. Our analysis also yielded new high-quality predictions of many functional site residues that were not catalogued in the curated data sources we inspected. We conclude that both DPA and text mining independently provide valuable high-throughput protein functional site predictions, and that integrating the two methods using LEAP-FS further improves the quality of these predictions. PMID:22393388

  16. Leaching From Biomass Gasification Residues

    DEFF Research Database (Denmark)

    Allegrini, Elisa; Boldrin, Alessio; Polletini, A.

    2011-01-01

    The aim of the present work is to attain an overall characterization of solid residues from biomass gasification. Besides the determination of chemical and physical properties, the work was focused on the study of leaching behaviour. Compliance and pH-dependence leaching tests coupled with geoche......The aim of the present work is to attain an overall characterization of solid residues from biomass gasification. Besides the determination of chemical and physical properties, the work was focused on the study of leaching behaviour. Compliance and pH-dependence leaching tests coupled...

  17. Carbaryl residues in maize products

    International Nuclear Information System (INIS)

    Zayed, S.M.A.D.; Mansour, S.A.; Mostafa, I.Y.; Hassan, A.

    1976-01-01

    The 14 C-labelled insecticide carbaryl was synthesized from [1- 14 C]-1-naphthol at a specific activity of 3.18mCig -1 . Maize plants were treated with the labelled insecticide under simulated conditions of agricultural practice. Mature plants were harvested and studied for distribution of total residues in untreated grains as popularly roasted and consumed, and in the corn oil and corn germ products. Total residues found under these conditions in the respective products were 0.2, 0.1, 0.45 and 0.16ppm. (author)

  18. Combinatorial construction of toric residues

    OpenAIRE

    Khetan, Amit; Soprounov, Ivan

    2004-01-01

    The toric residue is a map depending on n+1 semi-ample divisors on a complete toric variety of dimension n. It appears in a variety of contexts such as sparse polynomial systems, mirror symmetry, and GKZ hypergeometric functions. In this paper we investigate the problem of finding an explicit element whose toric residue is equal to one. Such an element is shown to exist if and only if the associated polytopes are essential. We reduce the problem to finding a collection of partitions of the la...

  19. Research items regarding seismic residual risk evaluation

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2013-08-15

    After learning the Fukushima Dai-ichi NPP severe accidents in 2011, the government investigation committee proposed the effective use of probabilistic safety assessment (PSA), and now it is required to establish new safety rules reflecting the results of probabilistic risk assessment (PRA) and proposed severe accident measures. Since the Seismic Design Guide has been revised on September 19, 2006, JNES has been discussing seismic PRA (Levels 1-3) methods to review licensees' residual risk assessment while preparing seismic PRA models. Meanwhile, new safety standards for light water reactors are to be issued and enforced on July 2013, which require the residual risk of tsunami, in addition to earthquakes, should be lowered as much as possible. The Fukushima accidents raised the problems related to risk assessment, e.g. approaches based on multi-hazard (earthquake and tsunami), multi-unit, multi-site, and equipment's common cause failure. This fiscal year, while performing seismic and/or tsunami PRA to work on these problems, JNES picked up the equipment whose failure greatly contribute to core damage, surveyed accident management measures on those equipment as well as effectiveness to reduce core damage probability. (author)

  20. Lead sorption-desorption from organic residues.

    Science.gov (United States)

    Duarte Zaragoza, Victor M; Carrillo, Rogelio; Gutierrez Castorena, Carmen M

    2011-01-01

    Sorption and desorption are mechanisms involved in the reduction of metal mobility and bioavailability in organic materials. Metal release from substrates is controlled by desorption. The capacity of coffee husk and pulp residues, vermicompost and cow manure to adsorb Pb2+ was evaluated. The mechanisms involved in the sorption process were also studied. Organic materials retained high concentrations of lead (up to 36,000 mg L(-1)); however, the mechanisms of sorption varied according to the characteristics of each material: degree of decomposition, pH, cation exchange capacity and percentage of organic matter. Vermicompost and manure removed 98% of the Pb from solution. Lead precipitated in manure and vermicompost, forming lead oxide (PbO) and lead ferrite (PbFe4O7). Adsorption isotherms did not fit to the typical Freundlich and Langmuir equations. Not only specific and non-specific adsorption was observed, but also precipitation and coprecipitation. Lead desorption from vermicompost and cow manure was less than 2%. For remediation of Pb-polluted sites, the application of vermicompost and manure is recommended in places with alkaline soils because Pb precipitation can be induced, whereas coffee pulp residue is recommended for acidic soils where Pb is adsorbed.

  1. Alternatives to crop residues for soil amendment

    OpenAIRE

    Powell, J.M.; Unger, P.W.

    1997-01-01

    Metadata only record In semiarid agroecosystems, crop residues can provide important benefits of soil and water conservation, nutrient cycling, and improved subsequent crop yields. However, there are frequently multiple competing uses for residues, including animal forage, fuel, and construction material. This chapter discusses the various uses of crop residues and examines alternative soil amendments when crop residues cannot be left on the soil.

  2. Residual Structures in Latent Growth Curve Modeling

    Science.gov (United States)

    Grimm, Kevin J.; Widaman, Keith F.

    2010-01-01

    Several alternatives are available for specifying the residual structure in latent growth curve modeling. Two specifications involve uncorrelated residuals and represent the most commonly used residual structures. The first, building on repeated measures analysis of variance and common specifications in multilevel models, forces residual variances…

  3. Computing Decoupled Residuals for Compact Disc Players

    DEFF Research Database (Denmark)

    Odgaard, Peter Fogh; Stoustrup, Jakob; Andersen, Palle

    2006-01-01

    a pair of residuals generated by Compact Disc Player. However, these residuals depend on the performance of position servos in the Compact Disc Player. In other publications of the same authors a pair of decoupled residuals is derived. However, the computation of these alternative residuals has been...

  4. Plant residues: short term effect on sulphate, borate, zinc and copper adsorption by an acid oxisol

    Directory of Open Access Journals (Sweden)

    Dias Ana Cristi Basile

    2003-01-01

    Full Text Available Laboratory experiments were carried out to examine the effects of plant residues on Cu, Zn, B and S adsorption by an acidic oxisol. The plant residues were: black oats (Avena strigosa, oil seed radish(Raphanus sativus, velvet beans (Stizolobium cinereum, and pigeon pea (Cajanus cajan collected at flowering stage. Plant residues increased Cu and Zn adsorptions and decreased B and S adsorptions. The results indicated that for short term effect plant residues decreased the availabilities of Cu and Zn through metal organic complex reactions and increased availabilities of S and B through competition with organic anions by the adsorption sites on soil.

  5. Remaining Sites Verification Package for the 600-233 Waste Site, Vertical Pipe Near 100-B Electrical Laydown Area. Attachment to Waste Site Reclassification Form 2005-041

    International Nuclear Information System (INIS)

    Carlson, R.A.

    2005-01-01

    The 600-233 waste site consisted of three small-diameter pipelines within the 600-232 waste site, including previously unknown diesel fuel supply lines discovered during site remediation. The 600-233 waste site has been remediated to achieve the remedial action objectives specified in the Remaining Sites ROD. The results of verification sampling show that residual contaminant concentrations do not preclude any future uses and allow for unrestricted use of shallow zone soils. The results also demonstrate that residual contaminant concentrations are protective of groundwater and the Columbia River

  6. RESRAD, Residual Radioactive Material Guideline Implementation

    International Nuclear Information System (INIS)

    1998-01-01

    This code system is designed to calculate site-specific residual radioactive material guidelines, and radiation dose and excess cancer risk to an on-site resident (maximally exposed individual). A guideline is a radionuclide concentration or level of radioactivity that is acceptable if a site is to be used without radiological restrictions. Guidelines are expressed as concentrations of residual radionuclides in soil. Soil is unconsolidated earth material, including rubble and debris that may be present. The guidelines are based on the following principles: (1) the total effective dose equivalent should not exceed 100 mrem/yr for all plausible land uses and 30 mrem/yr for current and likely future land uses and (2) doses should be kept as low as reasonably achievable (ALARA). Nine environmental pathways are considered: direct exposure, inhalation of dust and radon, and ingestion of plant foods, meat, milk, aquatic foods, soil, and water. CCC-0552/04: A - Description of program or function: RESRAD-BUILD Version 2.36 is a pathway analysis model designed to evaluate the potential radiological dose incurred by an individual who works or lives in a building contaminated with radioactive material. The radioactive material in the building structure can be released into the indoor air by mechanisms such as diffusion (radon gas), mechanical removal (decontamination activities), or erosion (removable surface contamination). In the June 1998 update, RESRAD was updated to Version 5.82 and RESRAD-BUILD was updated to version 2.36. The following notes highlight new features: RESRAD5.82 (4/30/98): - Allow plot data to be exported to tab-delimited text file - Corrected Installation problem to Windows 3.1 - Corrected plotting problem for soil guidelines RESRAD-BUILD2.36 (4/9/98): - Corrected problem with simultaneously changing number of wall regions and their parameters - Added OK button to uncertainty window - Made sure first uncertainty variable in added on first try See the

  7. Managing woodwaste: Yield from residue

    Energy Technology Data Exchange (ETDEWEB)

    Nielson, E. [LNS Services, Inc., North Vancouver, British Columbia (Canada); Rayner, S. [Pacific Waste Energy Inc., Burnaby, British Columbia (Canada)

    1993-12-31

    Historically, the majority of sawmill waste has been burned or buried for the sole purpose of disposal. In most jurisdictions, environmental legislation will prohibit, or render uneconomic, these practices. Many reports have been prepared to describe the forest industry`s residue and its environmental effect; although these help those looking for industry-wide or regional solutions, such as electricity generation, they have limited value for the mill manager, who has the on-hands responsibility for generation and disposal of the waste. If the mill manager can evaluate waste streams and break them down into their usable components, he can find niche market solutions for portions of the plant residue and redirect waste to poor/no-return, rather than disposal-cost, end uses. In the modern mill, residue is collected at the individual machine centre by waste conveyors that combine and mix sawdust, shavings, bark, etc. and send the result to the hog-fuel pile. The mill waste system should be analyzed to determine the measures that can improve the quality of residues and determine the volumes of any particular category before the mixing, mentioned above, occurs. After this analysis, the mill may find a niche market for a portion of its woodwaste.

  8. Residual stress in polyethylene pipes

    Czech Academy of Sciences Publication Activity Database

    Poduška, Jan; Hutař, Pavel; Kučera, J.; Frank, A.; Sadílek, J.; Pinter, G.; Náhlík, Luboš

    2016-01-01

    Roč. 54, SEP (2016), s. 288-295 ISSN 0142-9418 R&D Projects: GA MŠk LM2015069; GA MŠk(CZ) LQ1601 Institutional support: RVO:68081723 Keywords : polyethylene pipe * residual stress * ring slitting method * lifetime estimation Subject RIV: JL - Materials Fatigue, Friction Mechanics Impact factor: 2.464, year: 2016

  9. Solow Residuals Without Capital Stocks

    DEFF Research Database (Denmark)

    Burda, Michael C.; Severgnini, Battista

    2014-01-01

    We use synthetic data generated by a prototypical stochastic growth model to assess the accuracy of the Solow residual (Solow, 1957) as a measure of total factor productivity (TFP) growth when the capital stock in use is measured with error. We propose two alternative measurements based on curren...

  10. Solidification process for sludge residue

    International Nuclear Information System (INIS)

    Pearce, K.L.

    1998-01-01

    This report investigates the solidification process used at 100-N Basin to solidify the N Basin sediment and assesses the N Basin process for application to the K Basin sludge residue material. This report also includes a discussion of a solidification process for stabilizing filters. The solidified matrix must be compatible with the Environmental Remediation Disposal Facility acceptance criteria

  11. Leptogenesis and residual CP symmetry

    International Nuclear Information System (INIS)

    Chen, Peng; Ding, Gui-Jun; King, Stephen F.

    2016-01-01

    We discuss flavour dependent leptogenesis in the framework of lepton flavour models based on discrete flavour and CP symmetries applied to the type-I seesaw model. Working in the flavour basis, we analyse the case of two general residual CP symmetries in the neutrino sector, which corresponds to all possible semi-direct models based on a preserved Z 2 in the neutrino sector, together with a CP symmetry, which constrains the PMNS matrix up to a single free parameter which may be fixed by the reactor angle. We systematically study and classify this case for all possible residual CP symmetries, and show that the R-matrix is tightly constrained up to a single free parameter, with only certain forms being consistent with successful leptogenesis, leading to possible connections between leptogenesis and PMNS parameters. The formalism is completely general in the sense that the two residual CP symmetries could result from any high energy discrete flavour theory which respects any CP symmetry. As a simple example, we apply the formalism to a high energy S 4 flavour symmetry with a generalized CP symmetry, broken to two residual CP symmetries in the neutrino sector, recovering familiar results for PMNS predictions, together with new results for flavour dependent leptogenesis.

  12. Site Management Guide (Blue Book)

    International Nuclear Information System (INIS)

    2014-01-01

    The U.S. Department of Energy (Department) Office of Legacy Management (LM), established in 2003, manages the Department's postclosure responsibilities and ensures the future protection of human health and the environment. During World War II and the Cold War, the Federal government developed and operated a vast network of industrial facilities for the research, production, and testing of nuclear weapons, as well as other scientific and engineering research. These processes left a legacy of radioactive and chemical waste, environmental contamination, and hazardous facilities and materials at well over 100 sites. Since 1989, the Department has taken an aggressive accelerated cleanup approach to reduce risks and cut costs. At most Departmental sites undergoing cleanup, some residual hazards will remain at the time cleanup is completed due to financial and technical impracticality. However, the Department still has an obligation to protect human health and the environment after cleanup is completed. LM fulfills DOE's postclosure obligation by providing long-term management of postcleanup sites which do not have continuing missions. LM is also responsible for sites under the Formerly Utilized Sites Remedial Action Program (FUSRAP). Currently, the U.S. Army Corps of Engineers (USACE) is responsible for site surveys and remediation at FUSRAP sites. Once remediation is completed, LM becomes responsible for long-term management. LM also has responsibility for uranium processing sites addressed by Title II of the Uranium Mill Tailings Radiation Control Act (UMTRCA). UMTRCA Title II sites are sites that were commercially owned and are regulated under a U.S. Nuclear Regulatory Commission (NRC) license. For license termination, the owner must conduct an NRC-approved cleanup of any on-site radioactive waste remaining from former uranium ore-processing operations. The site owner must also provide full funding for inspections and, if necessary, ongoing maintenance. Once site

  13. Comparison of neutron and synchrotron diffraction measurements of residual stress in bead-on-plate weldments

    International Nuclear Information System (INIS)

    Paradowska, A.M.; Price, J.W.; Finlayson, T.R.; Lienert, U.; Ibrahim, R.

    2010-01-01

    This paper explores the use of neutron and synchrotron diffractions for the evaluation of residual stresses in welded components. It has been shown that it is possible to achieve very good agreement between the two independent diffraction techniques. This study shows the significance of the weld start and end sites on the residual strain/stress distribution. Quantitative evaluation of the residual stress development process for multibead weldments has been presented. Some measurements were also taken before and after postweld stress relieving to establish the reduction and redistribution of the residual stress. The detailed measurements of residual stress around the weld achieved in this work significantly improve the knowledge and understanding of residual stress in welded components.

  14. Radioactive material in residues of health services residues

    International Nuclear Information System (INIS)

    Costa R, A. Jr.; Recio, J.C.

    2006-01-01

    The work presents the operational actions developed by the one organ responsible regulator for the control of the material use radioactive in Brazil. Starting from the appearance of coming radioactive material of hospitals and clinical with services of nuclear medicine, material that that is picked up and transported in specific trucks for the gathering of residuals of hospital origin, and guided one it manufactures of treatment of residuals of services of health, where they suffer radiological monitoring before to guide them for final deposition in sanitary embankment, in the city of Sao Paulo, Brazil. The appearance of this radioactive material exposes a possible one violation of the norms that govern the procedures and practices in that sector in the country. (Author)

  15. Superfund Sites

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This layer represents active Superfund Sites published by the Environmental Protection Agency (EPA). These data were extracted from the Superfund Enterprise...

  16. Site development

    International Nuclear Information System (INIS)

    Noack, J.

    1975-01-01

    The subject of this paper is a general view over all necessary considerations to develop the site after it has been chosen and before starting with the construction of a nuclear power plant. (orig./RW) [de

  17. Site selection

    International Nuclear Information System (INIS)

    Olsen, C.W.

    1983-07-01

    The conditions and criteria for selecting a site for a nuclear weapons test at the Nevada Test Site are summarized. Factors considered are: (1) scheduling of drill rigs, (2) scheduling of site preparation (dirt work, auger hole, surface casing, cementing), (3) schedule of event (when are drill hole data needed), (4) depth range of proposed W.P., (5) geologic structure (faults, Pz contact, etc.), (6) stratigraphy (alluvium, location of Grouse Canyon Tuff, etc.), (7) material properties (particularly montmorillonite and CO 2 content), (8) water table depth, (9) potential drilling problems (caving), (10) adjacent collapse craters and chimneys, (11) adjacent expended but uncollapsed sites, (12) adjacent post-shot or other small diameter holes, (13) adjacent stockpile emplacement holes, (14) adjacent planned events (including LANL), (15) projected needs of Test Program for various DOB's and operational separations, and (16) optimal use of NTS real estate

  18. Subsurface characterization and geohydrologic site evaluation West Chestnut Ridge site

    International Nuclear Information System (INIS)

    1984-01-01

    The West Chestnut Ridge Site at the Oak Ridge National Laboratory is being considered for use as a repository for low-level radioactive waste. The purposes of this study were to provide a geohydrological characterization of the site for use in pathways analysis, and to provide preliminary geotechnical recommendations that would be used for development of a site utilization plan. Subsurface conditions were investigated at twenty locations and observation wells were installed. Field testing at each location included the Standard Penetration Test and permeability tests in soil and rock. A well pumping test was ocmpleted at one site. Laboratory testing included permeability, deformability, strength and compaction tests, as well as index and physical property tests. The field investigations showed that the subsurface conditions include residual soil overlying a weathered zone of dolomite which grades into relatively unweathered dolomite at depth. The thickness of residual soil is typically 80 ft (24 m) on the ridges, but can be as little as 10 ft (3 m) in the valleys. Trench excavations to depths of 30 ft (9 m) should not present serious slope stability problems above the water table. On-site soils can be used for liners or trench backfill but these soils may require moisture conditioning to achieve required densities. 19 figures, 8 tables

  19. Release behavior of triazine residues in stabilised contaminated soils

    International Nuclear Information System (INIS)

    Ying, G.G.; Kookana, R.S.; Mallavarpu, M.

    2005-01-01

    This paper reports the release behavior of two triazines (atrazine and simazine) in stabilised soils from a pesticide-contaminated site in South Australia. The soils were contaminated with a range of pesticides, especially with triazine herbicides. With multiple extractions of each soil sample with deionised water (eight in total), 15% of atrazine and 4% of simazine residues were recovered, resulting in very high concentrations of the two herbicides in leachate. The presence of small fractions of surfactants was found to further enhance the release of the residues. Methanol content up to 10% did not substantially influence the concentration of simazine and atrazine released. The study demonstrated that while the stabilisation of contaminated soil with particulate activated carbon (5%) and cement mix (15%) was effective in locking the residues of some pesticides, it failed to immobilise triazine herbicides residues completely. Given the higher water solubility of these herbicides than other compounds more effective strategies to immobilise their residues is needed. - Stabilisation of contaminated soil with a mix of activated carbon and cement may fail to immobilise some contaminants like triazines

  20. Systems for harvesting and handling cotton plant residue

    Energy Technology Data Exchange (ETDEWEB)

    Coates, W. [Univ. of Arizona, Tucson, AZ (United States)

    1993-12-31

    In the warmer regions of the United States, cotton plant residue must be buried to prevent it from serving as an overwintering site for insect pests such as the pink bollworm. Most of the field operations used to bury the residue are high energy consumers and tend to degrade soil structure, thereby increasing the potential for erosion. The residue is of little value as a soil amendment and consequently is considered a negative value biomass. A commercial system to harvest cotton plant residue would be of both economic and environmental benefit to cotton producers. Research has been underway at the University of Arizona since the spring of 1991 to develop a commercially viable system for harvesting cotton plant residue. Equipment durability, degree of densification, energy required, cleanliness of the harvested material, and ease of product handling and transport are some of the performance variables which have been measured. Two systems have proven superior. In both, the plants are pulled from the ground using an implement developed specifically for the purpose. In one system, the stalks are baled using a large round baler, while in the other the stalks are chopped with a forage harvester, and then made into packages using a cotton module maker. Field capacities, energy requirements, package density and durability, and ease of handling with commercially available equipment have been measured for both systems. Selection of an optimum system for a specific operation depends upon end use of the product, and upon equipment availability.

  1. Tyrosine Residues Regulate Multiple Nuclear Functions of P54nrb.

    Science.gov (United States)

    Lee, Ahn R; Hung, Wayne; Xie, Ning; Liu, Liangliang; He, Leye; Dong, Xuesen

    2017-04-01

    The non-POU-domain-containing octamer binding protein (NONO; also known as p54nrb) has various nuclear functions ranging from transcription, RNA splicing, DNA synthesis and repair. Although tyrosine phosphorylation has been proposed to account for the multi-functional properties of p54nrb, direct evidence on p54nrb as a phosphotyrosine protein remains unclear. To investigate the tyrosine phosphorylation status of p54nrb, we performed site-directed mutagenesis on the five tyrosine residues of p54nrb, replacing the tyrosine residues with phenylalanine or alanine, and immunoblotted for tyrosine phosphorylation. We then preceded with luciferase reporter assays, RNA splicing minigene assays, co-immunoprecipitation, and confocal microscopy to study the function of p54nrb tyrosine residues on transcription, RNA splicing, protein-protein interaction, and cellular localization. We found that p54nrb was not phosphorylated at tyrosine residues. Rather, it has non-specific binding affinity to anti-phosphotyrosine antibodies. However, replacement of tyrosine with phenylalanine altered p54nrb activities in transcription co-repression and RNA splicing in gene context-dependent fashions by means of differential regulation of p54nrb protein association with its interacting partners and co-regulators of transcription and splicing. These results demonstrate that tyrosine residues, regardless of phosphorylation status, are important for p54nrb function. J. Cell. Physiol. 232: 852-861, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  2. Analysis of fenbendazole residues in bovine milk by ELISA.

    Science.gov (United States)

    Brandon, David L; Bates, Anne H; Binder, Ronald G; Montague, William C; Whitehand, Linda C; Barker, Steven A

    2002-10-09

    Fenbendazole residues in bovine milk were analyzed by ELISAs using two monoclonal antibodies. One monoclonal antibody (MAb 587) bound the major benzimidazole anthelmintic drugs, including fenbendazole, oxfendazole, and fenbendazole sulfone. The other (MAb 591) was more specific for fenbendazole, with 13% cross-reactivity with the sulfone and no significant binding to the sulfoxide metabolite. The limit of detection of the ELISA method in the milk matrix was 7 ppb for MAb 587 and 3 ppb for MAb 591. Fenbendazole was administered in feed, drench, and paste form to three groups of dairy cattle. Milk was collected immediately before dosing and then every 12 h for 5 days. The ELISA indicated that residue levels varied widely among individual cows in each group. Fenbendazole levels peaked at approximately 12-24 h and declined rapidly thereafter. Metabolites were detected at much higher levels than the parent compound, peaked at approximately 24-36 h, and declined gradually. Residue levels were undetectable by 72 h. The ELISA data correlated well with the total residues determined by chromatographic analysis, but the use of the two separate ELISAs did not afford an advantage over ELISA with the single, broadly reactive MAb 587. The ELISA method could be used to flag high-residue samples in on-site monitoring of fenbendazole in milk and is a potential tool for studying drug pharmacokinetics.

  3. Hanford tank residual waste - Contaminant source terms and release models

    International Nuclear Information System (INIS)

    Deutsch, William J.; Cantrell, Kirk J.; Krupka, Kenneth M.; Lindberg, Michael L.; Jeffery Serne, R.

    2011-01-01

    Highlights: → Residual waste from five Hanford spent fuel process storage tanks was evaluated. → Gibbsite is a common mineral in tanks with high Al concentrations. → Non-crystalline U-Na-C-O-P ± H phases are common in the U-rich residual. → Iron oxides/hydroxides have been identified in all residual waste samples. → Uranium release is highly dependent on waste and leachant compositions. - Abstract: Residual waste is expected to be left in 177 underground storage tanks after closure at the US Department of Energy's Hanford Site in Washington State, USA. In the long term, the residual wastes may represent a potential source of contamination to the subsurface environment. Residual materials that cannot be completely removed during the tank closure process are being studied to identify and characterize the solid phases and estimate the release of contaminants from these solids to water that might enter the closed tanks in the future. As of the end of 2009, residual waste from five tanks has been evaluated. Residual wastes from adjacent tanks C-202 and C-203 have high U concentrations of 24 and 59 wt.%, respectively, while residual wastes from nearby tanks C-103 and C-106 have low U concentrations of 0.4 and 0.03 wt.%, respectively. Aluminum concentrations are high (8.2-29.1 wt.%) in some tanks (C-103, C-106, and S-112) and relatively low ( 2 -saturated solution, or a CaCO 3 -saturated water. Uranium release concentrations are highly dependent on waste and leachant compositions with dissolved U concentrations one or two orders of magnitude higher in the tests with high U residual wastes, and also higher when leached with the CaCO 3 -saturated solution than with the Ca(OH) 2 -saturated solution. Technetium leachability is not as strongly dependent on the concentration of Tc in the waste, and it appears to be slightly more leachable by the Ca(OH) 2 -saturated solution than by the CaCO 3 -saturated solution. In general, Tc is much less leachable (<10 wt.% of the

  4. French uranium mining sites remediation

    International Nuclear Information System (INIS)

    Roche, M.

    2002-01-01

    Following a presentation of the COGEMA's general policy for the remediation of uranium mining sites and the regulatory requirements, the current phases of site remediation operations are described. Specific operations for underground mines, open pits, milling facilities and confining the milled residues to meet long term public health concerns are detailed and discussed in relation to the communication strategies to show and explain the actions of COGEMA. A brief review of the current remediation situation at the various French facilities is finally presented. (author)

  5. Flanking signal and mature peptide residues influence signal peptide cleavage

    Directory of Open Access Journals (Sweden)

    Ranganathan Shoba

    2008-12-01

    Full Text Available Abstract Background Signal peptides (SPs mediate the targeting of secretory precursor proteins to the correct subcellular compartments in prokaryotes and eukaryotes. Identifying these transient peptides is crucial to the medical, food and beverage and biotechnology industries yet our understanding of these peptides remains limited. This paper examines the most common type of signal peptides cleavable by the endoprotease signal peptidase I (SPase I, and the residues flanking the cleavage sites of three groups of signal peptide sequences, namely (i eukaryotes (Euk (ii Gram-positive (Gram+ bacteria, and (iii Gram-negative (Gram- bacteria. Results In this study, 2352 secretory peptide sequences from a variety of organisms with amino-terminal SPs are extracted from the manually curated SPdb database for analysis based on physicochemical properties such as pI, aliphatic index, GRAVY score, hydrophobicity, net charge and position-specific residue preferences. Our findings show that the three groups share several similarities in general, but they display distinctive features upon examination in terms of their amino acid compositions and frequencies, and various physico-chemical properties. Thus, analysis or prediction of their sequences should be separated and treated as distinct groups. Conclusion We conclude that the peptide segment recognized by SPase I extends to the start of the mature protein to a limited extent, upon our survey of the amino acid residues surrounding the cleavage processing site. These flanking residues possibly influence the cleavage processing and contribute to non-canonical cleavage sites. Our findings are applicable in defining more accurate prediction tools for recognition and identification of cleavage site of SPs.

  6. Development of irradiation technique on controlling food contamination residue

    International Nuclear Information System (INIS)

    Liu Bin; Xiong Shanbai; Xiong Guangquan; Cheng Wei; Chen Yuxia; Liao Tao; Li Xin; Lin Ruotai

    2010-01-01

    The current state of the researches of irradiation technology on controlling food mycotoxin, pesticide, veterinary drugs and fishery drugs residue was summarized. And the degradation rate, mechanism, products and toxicities of food contamination were expatiated. The free radical from irradiation attack the site of weaker bond, and the less or more toxic substances were produced, which lead to the degradation of the food contamination. The limitations and future application of irradiation technique on controlling food contamination were also analyzed. (authors)

  7. The Cauchy method of residues

    CERN Document Server

    Mitrinović, Dragoslav S

    1993-01-01

    Volume 1, i. e. the monograph The Cauchy Method of Residues - Theory and Applications published by D. Reidel Publishing Company in 1984 is the only book that covers all known applications of the calculus of residues. They range from the theory of equations, theory of numbers, matrix analysis, evaluation of real definite integrals, summation of finite and infinite series, expansions of functions into infinite series and products, ordinary and partial differential equations, mathematical and theoretical physics, to the calculus of finite differences and difference equations. The appearance of Volume 1 was acknowledged by the mathematical community. Favourable reviews and many private communications encouraged the authors to continue their work, the result being the present book, Volume 2, a sequel to Volume 1. We mention that Volume 1 is a revised, extended and updated translation of the book Cauchyjev raeun ostataka sa primenama published in Serbian by Nau~na knjiga, Belgrade in 1978, whereas the greater part ...

  8. Residual life management. Maintenance improvement

    International Nuclear Information System (INIS)

    Sainero Garcia, J.; Hevia Ruperez, F.

    1995-01-01

    The terms Residual Life Management, Life Cycle Management and Long-Term Management are synonymous with a concept which aims to establish efficient maintenance for the profitable and safe operation of a power plant for as long as possible. A Residual Life Management programme comprises a number of stages, of which Maintenance Evaluation focuses on how power plant maintenance practices allow the mitigation and control of component ageing. with this objective in mind, a methodology has been developed for the analysis of potential degradative phenomena acting on critical components in terms of normal power plant maintenance practices. This methodology applied to maintenance evaluation enables the setting out of a maintenance programme based on the Life Management concept, and the programme's subsequent up-dating to allow for new techniques and methods. Initial applications have shown that although, in general terms, power plant maintenance is efficient, the way in which Residual Life Management is approached requires changes in maintenance practices. These changes range from modifications to existing inspection and surveillance methods or the establishment of new ones, to the monitoring of trends or the performance of additional studies, the purpose of which is to provide an accurate evaluation of the condition of the installations and the possibility of life extension. (Author)

  9. Site development

    International Nuclear Information System (INIS)

    Gaynor, R.K.

    1989-01-01

    Development of a low-level radioactive waste land disposal facility is little different than any industrial development of similar scope. Consideration must be made for normal business and operations management, security, facility maintenance, traffic control and necessary amenities for personnel. The item specific to the low-level waste site is the handling of radioactive waste materials and the regulatory and environmental protection procedures that must be planned for and accomodated in the site design and development. Each of these elements and the facility as a whole must be designed to be compatible with local land use plans, available transportation and support services, and the social and economic goals of the local community. Plans should also be made for quality control and orderly construction. This chapter deals with those aspects of the facility, its design and construction which are integral parts to the overall performance of the site

  10. Soil carbon sequestration and changes in fungal and bacterial biomass following incorporation of forest residues

    Science.gov (United States)

    Matt D. Busse; Felipe G. Sanchez; Alice W. Ratcliff; John R. Butnor; Emily A. Carter; Robert F. Powers

    2009-01-01

    Sequestering carbon (C) in forest soils can benefit site fertility and help offset greenhouse gas emissions. However, identifying soil conditions and forest management practices which best promote C accumulation remains a challenging task. We tested whether soil incorporation of masticated woody residues alters short-term C storage at forested sites in western and...

  11. Site Practice

    DEFF Research Database (Denmark)

    Wahedi, Haseebullah

    2016-01-01

    different practices in the construction phase. The research is based on an ethnographic study of a case in Denmark. The empirical data were collected through direct observations and semi-structured interviews with site managers, contract managers, foremen and craftsmen. Findings revealed...... that the construction phase comprises several communities and practices, leading to various uses of the drawings. The results indicated that the craftsmen used drawings to position themselves in the correct location, and that the site managers and contract managers used them as management tools and legal documents...

  12. Characterisation and management of concrete grinding residuals.

    Science.gov (United States)

    Kluge, Matt; Gupta, Nautasha; Watts, Ben; Chadik, Paul A; Ferraro, Christopher; Townsend, Timothy G

    2018-02-01

    Concrete grinding residue is the waste product resulting from the grinding, cutting, and resurfacing of concrete pavement. Potential beneficial applications for concrete grinding residue include use as a soil amendment and as a construction material, including as an additive to Portland cement concrete. Concrete grinding residue exhibits a high pH, and though not hazardous, it is sufficiently elevated that precautions need to be taken around aquatic ecosystems. Best management practices and state regulations focus on reducing the impact on such aquatic environment. Heavy metals are present in concrete grinding residue, but concentrations are of the same magnitude as typically recycled concrete residuals. The chemical composition of concrete grinding residue makes it a useful product for some soil amendment purposes at appropriate land application rates. The presence of unreacted concrete in concrete grinding residue was examined for potential use as partial replacement of cement in new concrete. Testing of Florida concrete grinding residue revealed no dramatic reactivity or improvement in mortar strength.

  13. FINITE ELEMENT MODEL FOR PREDICTING RESIDUAL ...

    African Journals Online (AJOL)

    FINITE ELEMENT MODEL FOR PREDICTING RESIDUAL STRESSES IN ... the transverse residual stress in the x-direction (σx) had a maximum value of 375MPa ... the finite element method are in fair agreement with the experimental results.

  14. Polychlorinated Biphenyls (PCB) Residue Effects Database

    Data.gov (United States)

    U.S. Environmental Protection Agency — The PCB Residue Effects (PCBRes) Database was developed to assist scientists and risk assessors in correlating PCB and dioxin-like compound residues with toxic...

  15. Functional Sites Induce Long-Range Evolutionary Constraints in Enzymes.

    Directory of Open Access Journals (Sweden)

    Benjamin R Jack

    2016-05-01

    Full Text Available Functional residues in proteins tend to be highly conserved over evolutionary time. However, to what extent functional sites impose evolutionary constraints on nearby or even more distant residues is not known. Here, we report pervasive conservation gradients toward catalytic residues in a dataset of 524 distinct enzymes: evolutionary conservation decreases approximately linearly with increasing distance to the nearest catalytic residue in the protein structure. This trend encompasses, on average, 80% of the residues in any enzyme, and it is independent of known structural constraints on protein evolution such as residue packing or solvent accessibility. Further, the trend exists in both monomeric and multimeric enzymes and irrespective of enzyme size and/or location of the active site in the enzyme structure. By contrast, sites in protein-protein interfaces, unlike catalytic residues, are only weakly conserved and induce only minor rate gradients. In aggregate, these observations show that functional sites, and in particular catalytic residues, induce long-range evolutionary constraints in enzymes.

  16. Site selection

    CERN Multimedia

    CERN PhotoLab

    1968-01-01

    To help resolve the problem of site selection for the proposed 300 GeV machine, the Council selected "three wise men" (left to right, J H Bannier of the Netherlands, A Chavanne of Switzerland and L K Boggild of Denmark).

  17. Site Restoration

    Energy Technology Data Exchange (ETDEWEB)

    Noynaert, L.; Bruggeman, A.; Cornelissen, R.; Massaut, V.; Rahier, A

    2001-04-01

    The objectives, the programme, and the achievements of the Site Restoration Department of SCK-CEN in 2000 are summarised. Main activities include the decommissioning of the BR3 PWR-reactor as well as other clean-up activities, projects on waste minimisation and activities related to the management of decommissioning projects. The department provides consultancy and services to external organisations.

  18. Site Restoration

    International Nuclear Information System (INIS)

    Noynaert, L.; Bruggeman, A.; Cornelissen, R.; Massaut, V.; Rahier, A.

    2001-01-01

    The objectives, the programme, and the achievements of the Site Restoration Department of SCK-CEN in 2000 are summarised. Main activities include the decommissioning of the BR3 PWR-reactor as well as other clean-up activities, projects on waste minimisation and activities related to the management of decommissioning projects. The department provides consultancy and services to external organisations

  19. Progress in stabilization of plutonium and residues since DNFSB recommendation 94-1

    International Nuclear Information System (INIS)

    Ball, J.M.; Dustin, D.F.

    1998-01-01

    There are approximately 100 metric tons of residues at the Rocky Flats Environmental Technology Site containing approximately 3 metric tons of plutonium. The residues are byproducts of past plutonium operations incinerator ash; pyrochemical salts; graphite; sand, slag, and crucible; and miscellaneous forms of combustibles, glass, metal, and sludges. In September 1993, a report was released (Reference 1) which identified concerns with the chemical stability of the residues and with the integrity of packaging. In May 1997, the Defense Nuclear Facility Safety Board published recommendation 94-1 citing a concern for the residue stability and requiring that the possibly unstable residues be processed within 3 years and all others within 5 years. A risk categorization scheme was developed which assigned a numerical risk to each residue type based on the probability and consequence of occurrence of failures associated with the hazards identified. The residues were ranked for priority of stabilization actions. Urgent concerns were resolved. All residue drums were vented to eliminate the potential for hydrogen and other explosive gas accumulation. Leaded rubber gloves and ion exchange resins were washed to eliminate the explosion potential. An aggressive characterization program was implemented to search for any additional safety or environmental concerns and to gain more definitive information concerning the choice of processes for stabilization and disposition of the residues. This paper provides background on the safety issues and summarizes recent characterization data. The residue processing and disposition plans, including schedule and cost, are also summarized in the paper. Finally, the paper addresses initiatives undertaken by Safe Sites of Colorado to accelerate the residue program

  20. 9 CFR 311.39 - Biological residues.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Biological residues. 311.39 Section... Biological residues. Carcasses, organs, or other parts of carcasses of livestock shall be condemned if it is determined that they are adulterated because of the presence of any biological residues. ...

  1. Cycling of grain legume residue nitrogen

    DEFF Research Database (Denmark)

    Jensen, E.S.

    1995-01-01

    Symbiotic nitrogen fixation by legumes is the main input of nitrogen in ecological agriculture. The cycling of N-15-labelled mature pea (Pisum sativum L.) residues was studied during three years in small field plots and lysimeters. The residual organic labelled N declined rapidly during the initial...... management methods in order to conserve grain legume residue N sources within the soil-plant system....

  2. Neutron residual stress measurements in linepipe

    International Nuclear Information System (INIS)

    Law, Michael; Gnaepel-Herold, Thomas; Luzin, Vladimir; Bowie, Graham

    2006-01-01

    Residual stresses in gas pipelines are generated by manufacturing and construction processes and may affect the subsequent pipe integrity. In the present work, the residual stresses in eight samples of linepipe were measured by neutron diffraction. Residual stresses changed with some coating processes. This has special implications in understanding and mitigating stress corrosion cracking, a major safety and economic problem in some gas pipelines

  3. Dissipation and Residue Level of Thifluzamide in Rice Field Ecosystem

    Directory of Open Access Journals (Sweden)

    Weitao Chen

    2015-01-01

    Full Text Available An efficient modified QuEChERS method combined with high performance liquid chromatography-tandem mass spectrometry detection (HPLC-MS/MS was established and evaluated for the residue analysis of thifluzamide in rice grain, husk, straw, seedling, paddy water, and soil. Thifluzamide residues were extracted with acetonitrile, cleaned up with primary secondary amine (PSA, and then determined by HPLC-MS/MS. The fortified recoveries were 76%–106% with RSDs of 3%–13%. The results of the supervised field trials at two experiment sites showed that thifluzamide dissipated rapidly in paddy fields, and the half-lives in paddy water, soil, and rice seedling were 0.3–0.6 d, 1.8–3.6 d, and 4.3–13.9 d, respectively. At harvest time, when the preharvest interval (PHI was set as 21 d, the final residues of thifluzamide in rice grains were below the maximum residue limit (MRL of 0.5 mg/kg set by Japan, whereas the final residues in rice husk and straw were still high (the highest value reached 1.36 mg/kg in rice husk and 0.83 mg/kg in rice straw. The results indicated that the highest residue in rice grain was 0.23 mg/kg when PHI was 21 d, and only 6.9–11.0% of acute risk quotient of thifluzamide was occupied by the dietary daily intake in Chinese population consuming rice.

  4. Cellulosic ethanol production from agricultural residues in Nigeria

    International Nuclear Information System (INIS)

    Iye, Edward; Bilsborrow, Paul

    2013-01-01

    Nigeria′s Biofuels Policy introduced in 2007 mandates a 10% blend (E10) of bioethanol with gasoline. This study investigates the potential for the development of a cellulosic ethanol industry based on the availability of agricultural residues and models the number of commercial processing facilities that could be sited in the six Geo-political zones. The potential for cellulosic ethanol production from agricultural residues in Nigeria is 7556 km 3 per annum exceeding the mandate of 10% renewable fuel required and providing the potential for 12 large- and 11 medium-scale processing facilities based on the use of a single feedstock. Cassava and yam peelings provided in excess of 80% of the process residues available with enough feedstock to supply 10 large-scale facilities with a fairly even distribution across the zones. Sorghum straw, millet straw and maize stalks represented 75% of the potential resource available from field residues with the potential to supply 2 large- and 7 medium-scale processing facilities, all of which would be located in the north of the country. When a multi-feedstock approach is used, this provides the potential for either 29 large- or 58 medium-scale facilities based on outputs of 250 and 125 km 3 per annum respectively. - Highlights: • Nigeria′s Biofuels Policy mandates a 10% blend of bioethanol with gasoline. • Total bioethanol production from agricultural residues was 7556 km 3 per annum. • Process residues offer the greatest potential accounting for 62% of production. • Nigeria has the potential for 12 large- and 11 medium scale commercial. • The use of mixed feedstocks significantly increases the potential for production

  5. Natural radioactivity in petroleum residues

    International Nuclear Information System (INIS)

    Gazineu, M.H.P.; Gazineu, M.H.P.; Hazin, C.A.; Hazin, C.A.

    2006-01-01

    The oil extraction and production industry generates several types of solid and liquid wastes. Scales, sludge and water are typical residues that can be found in such facilities and that can be contaminated with Naturally Occurring Radioactive Material (N.O.R.M.). As a result of oil processing, the natural radionuclides can be concentrated in such residues, forming the so called Technologically Enhanced Naturally Occurring Radioactive Material, or T.E.N.O.R.M.. Most of the radionuclides that appear in oil and gas streams belong to the 238 U and 232 Th natural series, besides 40 K. The present work was developed to determine the radionuclide content of scales and sludge generated during oil extraction and production operations. Emphasis was given to the quantification of 226 Ra, 228 Ra and 40 K since these radionuclides,are responsible for most of the external exposure in such facilities. Samples were taken from the P.E.T.R.O.B.R.A.S. unity in the State of Sergipe, in Northeastern Brazil. They were collected directly from the inner surface of water pipes and storage tanks, or from barrels stored in the waste storage area of the E and P unit. The activity concentrations for 226 Ra, 228 Ra and 40 K were determined by using an HP Ge gamma spectrometric system. The results showed concentrations ranging from 42.7 to 2,110.0 kBq/kg for 226 Ra, 40.5 to 1,550.0 kBq/kg for 228 Ra, and 20.6 to 186.6 kBq/kg for 40 K. The results highlight the importance of determining the activity concentration of those radionuclides in oil residues before deciding whether they should be stored or discarded to the environment. (authors)

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

    Science.gov (United States)

    Genkov, Todor; Spreitzer, Robert J

    2009-10-30

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

  7. Residual Liquefaction under Standing Waves

    DEFF Research Database (Denmark)

    Kirca, V.S. Ozgur; Sumer, B. Mutlu; Fredsøe, Jørgen

    2012-01-01

    This paper summarizes the results of an experimental study which deals with the residual liquefaction of seabed under standing waves. It is shown that the seabed liquefaction under standing waves, although qualitatively similar, exhibits features different from that caused by progressive waves....... The experimental results show that the buildup of pore-water pressure and the resulting liquefaction first starts at the nodal section and spreads towards the antinodal section. The number of waves to cause liquefaction at the nodal section appears to be equal to that experienced in progressive waves for the same...

  8. Process to recycle shredder residue

    Science.gov (United States)

    Jody, Bassam J.; Daniels, Edward J.; Bonsignore, Patrick V.

    2001-01-01

    A system and process for recycling shredder residue, in which separating any polyurethane foam materials are first separated. Then separate a fines fraction of less than about 1/4 inch leaving a plastics-rich fraction. Thereafter, the plastics rich fraction is sequentially contacted with a series of solvents beginning with one or more of hexane or an alcohol to remove automotive fluids; acetone to remove ABS; one or more of EDC, THF or a ketone having a boiling point of not greater than about 125.degree. C. to remove PVC; and one or more of xylene or toluene to remove polypropylene and polyethylene. The solvents are recovered and recycled.

  9. Residual Stresses In 3013 Containers

    International Nuclear Information System (INIS)

    Mickalonis, J.; Dunn, K.

    2009-01-01

    The DOE Complex is packaging plutonium-bearing materials for storage and eventual disposition or disposal. The materials are handled according to the DOE-STD-3013 which outlines general requirements for stabilization, packaging and long-term storage. The storage vessels for the plutonium-bearing materials are termed 3013 containers. Stress corrosion cracking has been identified as a potential container degradation mode and this work determined that the residual stresses in the containers are sufficient to support such cracking. Sections of the 3013 outer, inner, and convenience containers, in both the as-fabricated condition and the closure welded condition, were evaluated per ASTM standard G-36. The standard requires exposure to a boiling magnesium chloride solution, which is an aggressive testing solution. Tests in a less aggressive 40% calcium chloride solution were also conducted. These tests were used to reveal the relative stress corrosion cracking susceptibility of the as fabricated 3013 containers. Significant cracking was observed in all containers in areas near welds and transitions in the container diameter. Stress corrosion cracks developed in both the lid and the body of gas tungsten arc welded and laser closure welded containers. The development of stress corrosion cracks in the as-fabricated and in the closure welded container samples demonstrates that the residual stresses in the 3013 containers are sufficient to support stress corrosion cracking if the environmental conditions inside the containers do not preclude the cracking process.

  10. Residual Fragments after Percutaneous Nephrolithotomy

    Directory of Open Access Journals (Sweden)

    Kaan Özdedeli

    2012-09-01

    Full Text Available Clinically insignificant residual fragments (CIRFs are described as asymptomatic, noninfectious and nonobstructive stone fragments (≤4 mm remaining in the urinary system after the last session of any intervention (ESWL, URS or PCNL for urinary stones. Their insignificance is questionable since CIRFs could eventually become significant, as their presence may result in recurrent stone growth and they may cause pain and infection due to urinary obstruction. They may become the source of persistent infections and a significant portion of the patients will have a stone-related event, requiring auxilliary interventions. CT seems to be the ultimate choice of assessment. Although there is no concensus about the timing, recent data suggests that it may be performed one month after the procedure. However, imaging can be done in the immediate postoperative period, if there are no tubes blurring the assessment. There is some evidence indicating that selective medical therapy may have an impact on decreasing stone formation rates. Retrograde intrarenal surgery, with its minimally invasive nature, seems to be the best way to deal with residual fragments.

  11. Residual number processing in dyscalculia.

    Science.gov (United States)

    Cappelletti, Marinella; Price, Cathy J

    2014-01-01

    Developmental dyscalculia - a congenital learning disability in understanding numerical concepts - is typically associated with parietal lobe abnormality. However, people with dyscalculia often retain some residual numerical abilities, reported in studies that otherwise focused on abnormalities in the dyscalculic brain. Here we took a different perspective by focusing on brain regions that support residual number processing in dyscalculia. All participants accurately performed semantic and categorical colour-decision tasks with numerical and non-numerical stimuli, with adults with dyscalculia performing slower than controls in the number semantic tasks only. Structural imaging showed less grey-matter volume in the right parietal cortex in people with dyscalculia relative to controls. Functional MRI showed that accurate number semantic judgements were maintained by parietal and inferior frontal activations that were common to adults with dyscalculia and controls, with higher activation for participants with dyscalculia than controls in the right superior frontal cortex and the left inferior frontal sulcus. Enhanced activation in these frontal areas was driven by people with dyscalculia who made faster rather than slower numerical decisions; however, activation could not be accounted for by response times per se, because it was greater for fast relative to slow dyscalculics but not greater for fast controls relative to slow dyscalculics. In conclusion, our results reveal two frontal brain regions that support efficient number processing in dyscalculia.

  12. Residual number processing in dyscalculia

    Directory of Open Access Journals (Sweden)

    Marinella Cappelletti

    2014-01-01

    Full Text Available Developmental dyscalculia – a congenital learning disability in understanding numerical concepts – is typically associated with parietal lobe abnormality. However, people with dyscalculia often retain some residual numerical abilities, reported in studies that otherwise focused on abnormalities in the dyscalculic brain. Here we took a different perspective by focusing on brain regions that support residual number processing in dyscalculia. All participants accurately performed semantic and categorical colour-decision tasks with numerical and non-numerical stimuli, with adults with dyscalculia performing slower than controls in the number semantic tasks only. Structural imaging showed less grey-matter volume in the right parietal cortex in people with dyscalculia relative to controls. Functional MRI showed that accurate number semantic judgements were maintained by parietal and inferior frontal activations that were common to adults with dyscalculia and controls, with higher activation for participants with dyscalculia than controls in the right superior frontal cortex and the left inferior frontal sulcus. Enhanced activation in these frontal areas was driven by people with dyscalculia who made faster rather than slower numerical decisions; however, activation could not be accounted for by response times per se, because it was greater for fast relative to slow dyscalculics but not greater for fast controls relative to slow dyscalculics. In conclusion, our results reveal two frontal brain regions that support efficient number processing in dyscalculia.

  13. Residual number processing in dyscalculia☆

    Science.gov (United States)

    Cappelletti, Marinella; Price, Cathy J.

    2013-01-01

    Developmental dyscalculia – a congenital learning disability in understanding numerical concepts – is typically associated with parietal lobe abnormality. However, people with dyscalculia often retain some residual numerical abilities, reported in studies that otherwise focused on abnormalities in the dyscalculic brain. Here we took a different perspective by focusing on brain regions that support residual number processing in dyscalculia. All participants accurately performed semantic and categorical colour-decision tasks with numerical and non-numerical stimuli, with adults with dyscalculia performing slower than controls in the number semantic tasks only. Structural imaging showed less grey-matter volume in the right parietal cortex in people with dyscalculia relative to controls. Functional MRI showed that accurate number semantic judgements were maintained by parietal and inferior frontal activations that were common to adults with dyscalculia and controls, with higher activation for participants with dyscalculia than controls in the right superior frontal cortex and the left inferior frontal sulcus. Enhanced activation in these frontal areas was driven by people with dyscalculia who made faster rather than slower numerical decisions; however, activation could not be accounted for by response times per se, because it was greater for fast relative to slow dyscalculics but not greater for fast controls relative to slow dyscalculics. In conclusion, our results reveal two frontal brain regions that support efficient number processing in dyscalculia. PMID:24266008

  14. Residual radioactive contamination at Maralinga and Emu, 1985

    International Nuclear Information System (INIS)

    Lokan, K.H.

    1985-04-01

    An account is provided of residual contamination at Maralinga and Emu, in South Australia, where the United Kingdom Atomic Weapons Research Establishment conducted nuclear weapons development trials between 1953 and 1963. Detailed information is presented about contamination levels at sites on the range where radioactive materials were dispersed. Some of these were associated with trials involving natural uranium or short-lived isotopes which are no longer present. There are four sites where plutonium-239 was dispersed in substantial quantities from minor trials and information is presented about its distribution. Much of this material has been diluted by mixing with local soil, but there is a significant quantity of material present in the form of contaminated fragments, particularly at Taranaki. A considerable quantity of uranium-235 is also present at Taranaki. An assessment is made of the radiological significance of the dispersed plutonium and it is concluded that the material represents a potential long term hazard while it remains in its present form. Residual radioactivity associated with all but one of the seven major trial sites involving nuclear explosions continues to decay in a predictable way and will in the worst case, fall below levels considered safe for continuous occupancy within about fifty years. One site, Tadje, contains significant concentrations of plutonium over a small area and onsidered to be an additional plutonium-contaminated locality. Measurements of beryllium concentrations in soil are presented

  15. Effects of logging residue management on the growth and nutriend distribution of a pinus taeda plantation in central Louisiana, USA

    Science.gov (United States)

    A. Tiarks; M. Elliot-Smith; R. Stagg

    2004-01-01

    A 37-year-old pine plantation was harvested. An experiment was established at the site with three levels of logging residue retention and two levels of weed control. By age 10 years retaining harvest residue increased pine volumes by 10 m3 ha-land weed control increased production by another 20 m3 ha

  16. The myeloperoxidase-derived oxidant hypothiocyanous acid inhibits protein tyrosine phosphatases via oxidation of key cysteine residues

    DEFF Research Database (Denmark)

    Cook, Naomi L.; Moeke, Cassidy H.; Fantoni, Luca I.

    2016-01-01

    Phosphorylation of protein tyrosine residues is critical to cellular processes, and is regulated by kinases and phosphatases (PTPs). PTPs contain a redox-sensitive active site Cys residue, which is readily oxidized. Myeloperoxidase, released from activated leukocytes, catalyzes thiocyanate ion (SCN...

  17. Microwave calcination for plutonium immobilization and residue stabilization

    Energy Technology Data Exchange (ETDEWEB)

    Harris, M.J.; Rising, T.L.; Roushey, W.J.; Sprenger, G.S. [Kaiser-Hill Co., Golden, CO (United States)

    1995-12-01

    In the late 1980`s development was begun on a process using microwave energy to vitrify low level mixed waste sludge and transuranic mixed waste sludge generated in Building 374 at Rocky Flats. This process was shown to produce a dense, highly durable waste form. With the cessation of weapons production at Rocky Flats, the emphasis has changed from treatment of low level and TRU wastes to stabilizaiton of plutonium oxide and residues. This equipment is versatile and can be used as a heat source to calcine, react or vitrify many types of residues and oxides. It has natural economies in that it heats only the material to be treated, significantly reducing cycle times over conventional furnaces. It is inexpensive to operate in that most of the working components remain outside of any necessary contamination enclosure and therefore can easily be maintained. Limited testing has been successfully performed on cerium oxide (as a surrogate for plutonium oxide), surrogate electrorefining salts, surrogate residue sludge and residue ash. Future plans also include tests on ion exchange resins. In an attempt to further the usefullness of this technology, a mobile, self-contained microwave melting system is currently under development and expected to be operational at Rocky Flats Enviromental Technology Site by the 4th quarter of FY96.

  18. Detection of antibiotic residues in food by Charm II test

    International Nuclear Information System (INIS)

    Addali, Mohamed

    2014-01-01

    Antibiotics are used in food to: -therapy and prophylaxis, -increase the productivity of the food producing animals. The presence of antimicrobial residues: -constitutes a potential human health hazard. has significant impact on international food trade. has implications on technological process in dairy industry. Detection of antibiotic residues is of great interest. It helps protect humans against the effects of such residues, the more it can support the participation of our country in international trade. Charm II test is one of the methods of detection of antimicrobial residues. The tests utilize microbial or antibody receptor assay technology. The sample is incubated with a binding agent (microbial cells with specific receptor sites or with specific antibodies attached) and a tracer (the radio-labeled version of the antibiotic to be detected). The amount of tracer on the binding agent is measured using a scintillation counter and is compared to a pre-determined cut-off or control point. If contaminating antibiotic is present, it will prevent the binding of the tracer by occupying the receptors on the binding agent. The less labeled tracer detected, the more contaminating antibiotic there is present in the sample. This work, carried out at the Radiochemical Laboratory of the National Centre of Nuclear Science and Technology, has two parts: 1/ The first is reserved to a literature review provides an overview on antibiotics and the charm II method. 2/ The second is devoted to the experimental study and presentation of results.

  19. Microwave calcination for plutonium immobilization and residue stabilization

    International Nuclear Information System (INIS)

    Harris, M.J.; Rising, T.L.; Roushey, W.J.; Sprenger, G.S.

    1995-01-01

    In the late 1980's development was begun on a process using microwave energy to vitrify low level mixed waste sludge and transuranic mixed waste sludge generated in Building 374 at Rocky Flats. This process was shown to produce a dense, highly durable waste form. With the cessation of weapons production at Rocky Flats, the emphasis has changed from treatment of low level and TRU wastes to stabilizaiton of plutonium oxide and residues. This equipment is versatile and can be used as a heat source to calcine, react or vitrify many types of residues and oxides. It has natural economies in that it heats only the material to be treated, significantly reducing cycle times over conventional furnaces. It is inexpensive to operate in that most of the working components remain outside of any necessary contamination enclosure and therefore can easily be maintained. Limited testing has been successfully performed on cerium oxide (as a surrogate for plutonium oxide), surrogate electrorefining salts, surrogate residue sludge and residue ash. Future plans also include tests on ion exchange resins. In an attempt to further the usefullness of this technology, a mobile, self-contained microwave melting system is currently under development and expected to be operational at Rocky Flats Enviromental Technology Site by the 4th quarter of FY96

  20. IMPACT OF COSOLVENT FLUSHING ON SUBSURFACE MICROBIAL ECOLOGY AT THE FORMER SAGE'S DRY CLEANER SITE

    Science.gov (United States)

    The Solvent Extraction Residual Biotreatment (SERB) technology was evaluated at the former Sage's Dry Cleaner site in Jacksonville, FL where an area of tetrachloroethylene (PCE) contamination was identified. The SERB technology is a treatment train approach to complete site rest...

  1. Gas-Phase Hydrogen-Deuterium Exchange Labeling of Select Peptide Ion Conformer Types: a Per-Residue Kinetics Analysis.

    Science.gov (United States)

    Khakinejad, Mahdiar; Kondalaji, Samaneh Ghassabi; Tafreshian, Amirmahdi; Valentine, Stephen J

    2015-07-01

    The per-residue, gas-phase hydrogen deuterium exchange (HDX) kinetics for individual amino acid residues on selected ion conformer types of the model peptide KKDDDDDIIKIIK have been examined using ion mobility spectrometry (IMS) and HDX-tandem mass spectrometry (MS/MS) techniques. The [M + 4H](4+) ions exhibit two major conformer types with collision cross sections of 418 Å(2) and 446 Å(2); the [M + 3H](3+) ions also yield two different conformer types having collision cross sections of 340 Å(2) and 367 Å(2). Kinetics plots of HDX for individual amino acid residues reveal fast- and slow-exchanging hydrogens. The contributions of each amino acid residue to the overall conformer type rate constant have been estimated. For this peptide, N- and C-terminal K residues exhibit the greatest contributions for all ion conformer types. Interior D and I residues show decreased contributions. Several charge state trends are observed. On average, the D residues of the [M + 3H](3+) ions show faster HDX rate contributions compared with [M + 4H](4+) ions. In contrast the interior I8 and I9 residues show increased accessibility to exchange for the more elongated [M + 4H](4+) ion conformer type. The contribution of each residue to the overall uptake rate showed a good correlation with a residue hydrogen accessibility score model calculated using a distance from charge site and initial incorporation site for nominal structures obtained from molecular dynamic simulations (MDS).

  2. Decommissioning of Malargue site: Engineering solution

    International Nuclear Information System (INIS)

    Liseno, Aldo; Giordano, Nolberto L.

    2000-01-01

    This paper provides a brief comment related with a projected repository to managed tailings of Malargue Plant, located in the city of Malargue (Mendoza province). The approved plan for restoration of Malargue consist of on site relocation of tailings to an engineered repository. The Site is located too close of Malargue city. To isolate tailings from the environment a multilayer system has been projected, in order to keep humidity of residues and multilayer, to avoid migration of contaminant to the environment. (author)

  3. Chloride binding site of neurotransmitter sodium symporters

    DEFF Research Database (Denmark)

    Kantcheva, Adriana Krassimirova; Quick, Matthias; Shi, Lei

    2013-01-01

    Neurotransmitter:sodium symporters (NSSs) play a critical role in signaling by reuptake of neurotransmitters. Eukaryotic NSSs are chloride-dependent, whereas prokaryotic NSS homologs like LeuT are chloride-independent but contain an acidic residue (Glu290 in LeuT) at a site where eukaryotic NSSs...

  4. MORTAR WITH UNSERVICEABLE TIRE RESIDUES

    Directory of Open Access Journals (Sweden)

    J. A. Canova

    2009-01-01

    Full Text Available This study analyzes the effects of unserviceable tire residues on rendering mortar using lime and washed sand at a volumetric proportion of 1:6. The ripened composite was dried in an oven and combined with both cement at a volumetric proportion of 1:1.5:9 and rubber powder in proportional aggregate volumes of 6, 8, 10, and 12%. Water exudation was evaluated in the plastic state. Water absorption by capillarity, fresh shrinkage and mass loss, restrained shrinkage and mass loss, void content, flexural strength, and deformation energy under compression were evaluated in the hardened state. There was an improvement in the water exudation and water absorption by capillarity and drying shrinkage, as well as a reduction of the void content and flexural strength. The product studied significantly aided the water exudation from mortar and, capillary elevation in rendering.

  5. MORTAR WITH UNSERVICEABLE TIRE RESIDUES

    Directory of Open Access Journals (Sweden)

    José Aparecido Canova

    2009-12-01

    Full Text Available This study analyzes the effects of unserviceable tire residues on rendering mortar using lime and washed sand at a volumetric proportion of 1:6. The ripened composite was dried in an oven and combined with both cement at a volumetric proportion of 1:1.5:9 and rubber powder in proportional aggregate volumes of 6, 8, 10, and 12%. Water exudation was evaluated in the plastic state. Water absorption by capillarity, fresh shrinkage and mass loss, restrained shrinkage and mass loss, void content, flexural strength, and deformation energy under compression were evaluated in the hardened state. There was an improvement in the water exudation and water absorption by capillarity and drying shrinkage, as well as a reduction of the void content and flexural strength. The product studied significantly aided the water exudation from mortar and, capillary elevation in rendering.

  6. Upgraded wood residue fuels 1995

    International Nuclear Information System (INIS)

    Vinterbaeck, J.

    1995-01-01

    The Swedish market for upgraded residue fuels, i.e. briquettes, pellets and wood powder, has developed considerably during the nineties. The additional costs for the upgrading processes are regained and create a surplus in other parts of the system, e.g. in the form of higher combustion efficiencies, lower investment costs for burning equipment, lower operation costs and a diminished environmental impact. All these factors put together have resulted in a rapid growth of this part of the energy sector. In 1994 the production was 1.9 TWh, an increase of 37 % compared to the previous year. In the forthcoming heating season 1995/96 the production may reach 4 TWh. 57 refs, 11 figs, 6 tabs

  7. Forest residues in cattle feed

    Directory of Open Access Journals (Sweden)

    João Elzeário Castelo Branco Iapichini

    2012-12-01

    Full Text Available The ruminants are capable of converting low-quality food, when they are complementes with high-energy source. Through the use of regional agricultural residues is possible to conduct more economical production systems, since energetic foods have high cost in animal production. There is very abundant availability of residues in agroforestry activities worldwide, so that if a small fraction of them were used with appropriate technical criteria they could largely meet the needs of existing herds in the world and thus meet the demands of consumption of protein of animal origin. The Southwest Region of São Paulo State has large area occupied by reforestation and wide availability of non-timber forest residues, which may represent more concentrated energetic food for ruminant production. This experiment aimed to evaluate the acceptability of ground pine (20, 30 and 40%, replacing part of the energetic food (corn, present in the composition of the concentrate and was performed at the Experimental Station of Itapetininga - Forest Institute / SMA, in the dry season of 2011. It were used four crossbred steers, mean 18 months old, average body weight of 250 kg, housed in a paddock provided with water ad libitum and covered troughs for supplementation with the experimental diet. The adjustment period of the animals was of 07 days and the measurement of the levels of consumption, physiological changes, acceptability and physiological parameters were observed during the following 25 days. The concentrate supplement was formulated based on corn (76.2%, Soybean Meal (20%, urea (2%, Ammonium sulfate (0.4%, calcite (1.4%, Mineral Core (1% and finely ground Pine Cone, replacing corn. In preparing food, the formulas were prepared to make them isoproteic/energetic, containing the following nutrient levels: 22% Crude Protein (CP and 79% of Total Nutrients (TDN. The animals received the supplement in three steps for each level of cone replaced, being offered in the

  8. Landfill Mining of Shredder Residues

    DEFF Research Database (Denmark)

    Hansen, Jette Bjerre; Hyks, Jiri; Shabeer Ahmed, Nassera

    In Denmark, shredder residues (SR) are classified as hazardous waste and until January 2012 the all SR were landfilled. It is estimated that more than 1.8 million tons of SR have been landfilled in mono cells. This paper describes investigations conducted at two Danish landfills. SR were excavated...... from the landfills and size fractionated in order to recover potential resources such as metal and energy and to reduce the amounts of SR left for re-landfilling. Based on the results it is estimated that 60-70% of the SR excavated could be recovered in terms of materials or energy. Only a fraction...... with particle size less than 5 mm needs to be re-landfilled at least until suitable techniques are available for recovery of materials with small particle sizes....

  9. Computational learning on specificity-determining residue-nucleotide interactions

    KAUST Repository

    Wong, Ka-Chun; Li, Yue; Peng, Chengbin; Moses, Alan M.; Zhang, Zhaolei

    2015-01-01

    The protein–DNA interactions between transcription factors and transcription factor binding sites are essential activities in gene regulation. To decipher the binding codes, it is a long-standing challenge to understand the binding mechanism across different transcription factor DNA binding families. Past computational learning studies usually focus on learning and predicting the DNA binding residues on protein side. Taking into account both sides (protein and DNA), we propose and describe a computational study for learning the specificity-determining residue-nucleotide interactions of different known DNA-binding domain families. The proposed learning models are compared to state-of-the-art models comprehensively, demonstrating its competitive learning performance. In addition, we describe and propose two applications which demonstrate how the learnt models can provide meaningful insights into protein–DNA interactions across different DNA binding families.

  10. Computational learning on specificity-determining residue-nucleotide interactions

    KAUST Repository

    Wong, Ka-Chun

    2015-11-02

    The protein–DNA interactions between transcription factors and transcription factor binding sites are essential activities in gene regulation. To decipher the binding codes, it is a long-standing challenge to understand the binding mechanism across different transcription factor DNA binding families. Past computational learning studies usually focus on learning and predicting the DNA binding residues on protein side. Taking into account both sides (protein and DNA), we propose and describe a computational study for learning the specificity-determining residue-nucleotide interactions of different known DNA-binding domain families. The proposed learning models are compared to state-of-the-art models comprehensively, demonstrating its competitive learning performance. In addition, we describe and propose two applications which demonstrate how the learnt models can provide meaningful insights into protein–DNA interactions across different DNA binding families.

  11. Residues and duality for projective algebraic varieties

    CERN Document Server

    Kunz, Ernst; Dickenstein, Alicia

    2008-01-01

    This book, which grew out of lectures by E. Kunz for students with a background in algebra and algebraic geometry, develops local and global duality theory in the special case of (possibly singular) algebraic varieties over algebraically closed base fields. It describes duality and residue theorems in terms of K�hler differential forms and their residues. The properties of residues are introduced via local cohomology. Special emphasis is given to the relation between residues to classical results of algebraic geometry and their generalizations. The contribution by A. Dickenstein gives applications of residues and duality to polynomial solutions of constant coefficient partial differential equations and to problems in interpolation and ideal membership. D. A. Cox explains toric residues and relates them to the earlier text. The book is intended as an introduction to more advanced treatments and further applications of the subject, to which numerous bibliographical hints are given.

  12. Remaining Sites Verification Package for the 100-D-2 Lead Sheeting Waste Site. Attachment to Waste Site Reclassification Form 2007-030

    International Nuclear Information System (INIS)

    Dittmer, L.M.

    2008-01-01

    The 100-D-2 Lead Sheeting waste site was located approximately 50 m southwest of the 185-D Building and approximately 16 m north of the east/west oriented road. The site consisted of a lead sheet covering a concrete pad. In accordance with this evaluation, the verification sampling results support a reclassification of this site to Interim Closed Out. The results of verification sampling show that residual contaminant concentrations do not preclude any future uses and allow for unrestricted use of shallow zone soils. The results also demonstrate that residual contaminant concentrations are protective of groundwater and the Columbia River

  13. Natively oxidized amino acid residues in the spinach cytochrome b 6 f complex.

    Science.gov (United States)

    Taylor, Ryan M; Sallans, Larry; Frankel, Laurie K; Bricker, Terry M

    2018-01-29

    The cytochrome b 6 f complex of oxygenic photosynthesis produces substantial levels of reactive oxygen species (ROS). It has been observed that the ROS production rate by b 6 f is 10-20 fold higher than that observed for the analogous respiratory cytochrome bc 1 complex. The types of ROS produced (O 2 •-, 1 O 2 , and, possibly, H 2 O 2 ) and the site(s) of ROS production within the b 6 f complex have been the subject of some debate. Proposed sources of ROS have included the heme b p , PQ p •- (possible sources for O 2 •- ), the Rieske iron-sulfur cluster (possible source of O 2 •- and/or 1 O 2 ), Chl a (possible source of 1 O 2 ), and heme c n (possible source of O 2 •- and/or H 2 O 2 ). Our working hypothesis is that amino acid residues proximal to the ROS production sites will be more susceptible to oxidative modification than distant residues. In the current study, we have identified natively oxidized amino acid residues in the subunits of the spinach cytochrome b 6 f complex. The oxidized residues were identified by tandem mass spectrometry using the MassMatrix Program. Our results indicate that numerous residues, principally localized near p-side cofactors and Chl a, were oxidatively modified. We hypothesize that these sites are sources for ROS generation in the spinach cytochrome b 6 f complex.

  14. Site Restoration

    Energy Technology Data Exchange (ETDEWEB)

    Noynaert, L.; Bruggeman, A.; Cornelissen, R.; Massaut, V.; Rahier, A

    2002-04-01

    The objectives, the programme, and the achievements of SCK-CEN's Site Restoration Department for 2001 are described. Main activities include the decommissioning of the BR3 PWR-reactor as well as other clean-up activities, projects on waste minimisation and the management of spent fuel and the flow of dismantled materials and the recycling of materials from decommissioning activities based on the smelting of metallic materials in specialised foundries. The department provides consultancy and services to external organisations and performs R and D on new techniques including processes for the treatment of various waste components including the reprocessing of spent fuel, the treatment of tritium, the treatment of liquid alkali metals into cabonates through oxidation, the treatment of radioactive organic waste and the reconditioning of bituminised waste products.

  15. Mochovce site

    International Nuclear Information System (INIS)

    1997-01-01

    In Mochovce site the construction of four units of WWER 440 NPP with V-213 type of reactor is being carried out. The financing of Mochovce units completion was resolved in April 1996. The completion work commenced at the construction site under leadership of SKODA Prague, the general supplier. The completion work on building part and tests of constructional electric distributions and lightning constructors started. The revisions in technological part were finished, and final protocols from revisions are the basis for starting of completion work. The assembly of transport container anchorage,ventilation system in hermetic areas and hermetic coverage of pools for stored spent nuclear fuel is being carried out. The pre-completion tests of instrumentation and control of ventilation systems, individual dosimetric control in medical station, and tests of nuclear programme according to commissioning and assembling work schedule at the equipment for physical protection of the NPP area started. Inspection activities at Mochovce were performed in accordance with inspection plan for 1996. Evaluation of routine inspections was performed by means of quarterly protocols. Main findings from the inspections performed in Mochovce were in the following areas: (a) deficiencies in the knowledge of the respective regulation and conditions from the Resolution of the state regulatory body, concerning selected employees; (b) training of the selected employees; (c) aim of the measures imposes by inspectors is to eliminate deficiencies in preparation of programmes for pre-completion and completion testing. NRA SR assessment activities at Mochovce NPP were focused mainly on approving and inspecting of design modification to approving programmes for pre-completion and completion testing of system s and equipment and on approving quality assurance programmes. The suggestions of international missions, which reviewed Mochovce safety in the years, were taken into consideration in the programme

  16. Distribution of residues and primitive roots

    Indian Academy of Sciences (India)

    Replacing the function f by g, we get the required estimate for N(p, N). D. Proof of Theorem 1.1. When p = 7, we clearly see that (1, 2) is a consecutive pair of quadratic residue modulo 7. Assume that p ≥ 11. If 10 is a quadratic residue modulo p, then we have (9, 10) as a consecutive pair of quadratic residues modulo p, ...

  17. Residual analysis for spatial point processes

    DEFF Research Database (Denmark)

    Baddeley, A.; Turner, R.; Møller, Jesper

    We define residuals for point process models fitted to spatial point pattern data, and propose diagnostic plots based on these residuals. The techniques apply to any Gibbs point process model, which may exhibit spatial heterogeneity, interpoint interaction and dependence on spatial covariates. Ou...... or covariate effects. Q-Q plots of the residuals are effective in diagnosing interpoint interaction. Some existing ad hoc statistics of point patterns (quadrat counts, scan statistic, kernel smoothed intensity, Berman's diagnostic) are recovered as special cases....

  18. Carbaryl residues in maize and processed products

    International Nuclear Information System (INIS)

    Qureshi, M.J.; Sattar, A. Jr.; Naqvi, M.H.

    1981-01-01

    Carbaryl residues in two local maize varieties were determined using a colorimetric method. No significant differences were observed for residues of the two varieties which ranged between 12.0 to 13.75 mg/kg in the crude oil, and averaged 1.04 and 0.67 mg/kg in the flour and cake respectively. In whole maize plants, carbaryl residues declined to approximately 2 mg/kg 35 days after treatment. Cooking in aqueous, oil or aqueous-oil media led to 63-83% loss of carbaryl residues, after 30 minutes. (author)

  19. Biogas and mineral fertiliser production from plant residues of phytoremediation

    Energy Technology Data Exchange (ETDEWEB)

    Chu, Thi Thu Ha

    2011-07-01

    The former uranium mining site in Ronneburg, Thuringia, Germany was known as a big source of uranium with more than 113,000 tons of uranium mined from 1946 to 1990. This area has been remediated since the nineties of the last century. However, nowadays the site in Ronneburg is still specially considered because of the heterogeneous contamination by many heavy metals and the vegetation can be affected. Three plant species including Indian mustard - Brassica juncea L., triticale - x. Triticosecale Wittmaek and sunflower - Helianthus annuus L. were seeded as accumulators of heavy metals and radionuclides in the phytoremediation process in 2009 and 2010 in Ronneburg. The subsequent utilization of the plant residues after phytoremediation is of special consideration. Batch fermentation of harvested plant materials under the mesophilic condition showed that all of the investigated plant materials had much higher biogas production than liquid cow manure except triticale root, of which biogas yield per volatile solid was not significantly higher than the one of sludge. The highest biogas yields (311 L{sub N}/kg FM and 807 L{sub N}/kg VS) were achieved from the spica of triticale after 42 days of retention of anaerobic digestion. Triticale shoot residues generated higher biogas and methane yields than the previously reported triticale materials that were harvested from the uncontaminated soil Triticale was considered as the highest potential species in biogas production, beside the best growth ability on the acidic soil at the test field site with highest biomass production. Biogas yield of Indian mustard shoot was also high but dramatically varied from 2009 to 2010. Digestates after anaerobic digestion of plant residues contained various macronutrients such as nitrogen, potassium, phosphorus and sulphur, and various micronutrients such as iron, manganes, zinc, etc. The accumulation levels of heavy metals in the investigated plant materials were not the hindrance factors

  20. Report on residual radioactive materials on public or acquired lands of the United States

    Energy Technology Data Exchange (ETDEWEB)

    None

    1979-07-01

    This report identifies sites located on public or acquired lands of the United States containing residual radioactive materials and other radioactive waste (excluding waste resulting from the production of electric energy) and was developed in accordance with the provisions of Section 114(b) of Public Law 95-604, "Uranium Mill Tailings Radiation Control Act of 1978," enacted on November 8, 1978. Additionally, the report specifies which Federal agency has jurisdiction over such sites and, where appropriate data were available, provides a description of the radiological status of each of the sites reported. For purpose of providing a timely report t o t h e Congress, a termination date of May 31, 1979 was established for the receipt, correlation, and analysis of the input data. As of this date, residual radioactive materials and other radioactive waste have been identified by six Federal agencies at 48 sites throughout the United States. Table 1 on page vi provides a summary listing of the number of sites under the jurisdiction of each of these reporting agencies. A cross listing in tabular form by affected state is presented in Table 2 on page viii. Of the 48 sites reported, 36 are located i n three western states - Colorado (27 sites), Wyoming (5 sites), and Utah (4 sites). Based upon t h e data submitted, the sites were categorized into three broad radiological status categories -- controlled, unstabilized, and risk to the public. At controlled sites, the residue is stabilized, access t o t h e site is controlled, the s i t e is well monitored, and does not currently constitute a risk to the public. At sites in the unstabilized category, a probability exists for the spread of contamination. Sites in the risk category contain residue which represents a long-term risk to the public under present conditions. Of the 48 reported sites,. 9 (approximately 19%) could be classified in the controlled category; 38 (approximately 79%) were in the unstabilized category and

  1. Computer Simulations Reveal Multiple Functions for Aromatic Residues in Cellulase Enzymes (Fact Sheet)

    Energy Technology Data Exchange (ETDEWEB)

    2012-07-01

    NREL researchers use high-performance computing to demonstrate fundamental roles of aromatic residues in cellulase enzyme tunnels. National Renewable Energy Laboratory (NREL) computer simulations of a key industrial enzyme, the Trichoderma reesei Family 6 cellulase (Cel6A), predict that aromatic residues near the enzyme's active site and at the entrance and exit tunnel perform different functions in substrate binding and catalysis, depending on their location in the enzyme. These results suggest that nature employs aromatic-carbohydrate interactions with a wide variety of binding affinities for diverse functions. Outcomes also suggest that protein engineering strategies in which mutations are made around the binding sites may require tailoring specific to the enzyme family. Cellulase enzymes ubiquitously exhibit tunnels or clefts lined with aromatic residues for processing carbohydrate polymers to monomers, but the molecular-level role of these aromatic residues remains unknown. In silico mutation of the aromatic residues near the catalytic site of Cel6A has little impact on the binding affinity, but simulation suggests that these residues play a major role in the glucopyranose ring distortion necessary for cleaving glycosidic bonds to produce fermentable sugars. Removal of aromatic residues at the entrance and exit of the cellulase tunnel, however, dramatically impacts the binding affinity. This suggests that these residues play a role in acquiring cellulose chains from the cellulose crystal and stabilizing the reaction product, respectively. These results illustrate that the role of aromatic-carbohydrate interactions varies dramatically depending on the position in the enzyme tunnel. As aromatic-carbohydrate interactions are present in all carbohydrate-active enzymes, the results have implications for understanding protein structure-function relationships in carbohydrate metabolism and recognition, carbon turnover in nature, and protein engineering

  2. Structural basis of carbohydrate recognition by lectin II from Ulex europaeus, a protein with a promiscuous carbohydrate-binding site.

    Science.gov (United States)

    Loris, R; De Greve, H; Dao-Thi, M H; Messens, J; Imberty, A; Wyns, L

    2000-08-25

    Protein-carbohydrate interactions are the language of choice for inter- cellular communication. The legume lectins form a large family of homologous proteins that exhibit a wide variety of carbohydrate specificities. The legume lectin family is therefore highly suitable as a model system to study the structural principles of protein-carbohydrate recognition. Until now, structural data are only available for two specificity families: Man/Glc and Gal/GalNAc. No structural data are available for any of the fucose or chitobiose specific lectins. The crystal structure of Ulex europaeus (UEA-II) is the first of a legume lectin belonging to the chitobiose specificity group. The complexes with N-acetylglucosamine, galactose and fucosylgalactose show a promiscuous primary binding site capable of accommodating both N-acetylglucos amine or galactose in the primary binding site. The hydrogen bonding network in these complexes can be considered suboptimal, in agreement with the low affinities of these sugars. In the complexes with chitobiose, lactose and fucosyllactose this suboptimal hydrogen bonding network is compensated by extensive hydrophobic interactions in a Glc/GlcNAc binding subsite. UEA-II thus forms the first example of a legume lectin with a promiscuous binding site and illustrates the importance of hydrophobic interactions in protein-carbohydrate complexes. Together with other known legume lectin crystal structures, it shows how different specificities can be grafted upon a conserved structural framework. Copyright 2000 Academic Press.

  3. Process for measuring residual stresses

    International Nuclear Information System (INIS)

    Elfinger, F.X.; Peiter, A.; Theiner, W.A.; Stuecker, E.

    1982-01-01

    No single process can at present solve all problems. The complete destructive processes only have a limited field of application, as the component cannot be reused. However, they are essential for the basic determination of stress distributions in the field of research and development. Destructive and non-destructive processes are mainly used if investigations have to be carried out on original components. With increasing component size, the part of destructive tests becomes smaller. The main applications are: quality assurance, testing of manufactured parts and characteristics of components. Among the non-destructive test procedures, X-raying has been developed most. It gives residual stresses on the surface and on surface layers near the edges. Further development is desirable - in assessment - in measuring techniques. Ultrasonic and magnetic crack detection processes are at present mainly used in research and development, and also in quality assurance. Because of the variable depth of penetration and the possibility of automation they are gaining in importance. (orig./RW) [de

  4. Residuals Management and Water Pollution Control Planning.

    Science.gov (United States)

    Environmental Protection Agency, Washington, DC. Office of Public Affairs.

    This pamphlet addresses the problems associated with residuals and water quality especially as it relates to the National Water Pollution Control Program. The types of residuals and appropriate management systems are discussed. Additionally, one section is devoted to the role of citizen participation in developing management programs. (CS)

  5. Tank 12H residuals sample analysis report

    Energy Technology Data Exchange (ETDEWEB)

    Oji, L. N. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL); Shine, E. P. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL); Diprete, D. P. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL); Coleman, C. J. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL); Hay, M. S. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2015-06-11

    The Savannah River National Laboratory (SRNL) was requested by Savannah River Remediation (SRR) to provide sample preparation and analysis of the Tank 12H final characterization samples to determine the residual tank inventory prior to grouting. Eleven Tank 12H floor and mound residual material samples and three cooling coil scrape samples were collected and delivered to SRNL between May and August of 2014.

  6. Soil water evaporation and crop residues

    Science.gov (United States)

    Crop residues have value when left in the field and also when removed from the field and sold as a commodity. Reducing soil water evaporation (E) is one of the benefits of leaving crop residues in place. E was measured beneath a corn canopy at the soil suface with nearly full coverage by corn stover...

  7. Densification of FL Chains via Residuated Frames

    Czech Academy of Sciences Publication Activity Database

    Baldi, Paolo; Terui, K.

    2016-01-01

    Roč. 75, č. 2 (2016), s. 169-195 ISSN 0002-5240 R&D Projects: GA ČR GAP202/10/1826 Keywords : densifiability * standard completeness * residuated lattices * residuated frames * fuzzy logic Subject RIV: BA - General Mathematics Impact factor: 0.625, year: 2016

  8. Does Bt Corn Really Produce Tougher Residues

    Science.gov (United States)

    Bt corn hybrids produce insecticidal proteins that are derived from a bacterium, Bacillus thuringiensis. There have been concerns that Bt corn hybrids produce residues that are relatively resistant to decomposition. We conducted four experiments that examined the decomposition of corn residues und...

  9. Semantic Tagging with Deep Residual Networks

    NARCIS (Netherlands)

    Bjerva, Johannes; Plank, Barbara; Bos, Johan

    2016-01-01

    We propose a novel semantic tagging task, semtagging, tailored for the purpose of multilingual semantic parsing, and present the first tagger using deep residual networks (ResNets). Our tagger uses both word and character representations and includes a novel residual bypass architecture. We evaluate

  10. Cement production from coal conversion residues

    International Nuclear Information System (INIS)

    Brown, L.D.; Clavenna, L.R.; Eakman, J.M.; Nahas, N.C.

    1981-01-01

    Cement is produced by feeding residue solids containing carbonaceous material and ash constituents obtained from converting a carbonaceous feed material into liquids and/or gases into a cement-making zone and burning the carbon in the residue solids to supply at least a portion of the energy required to convert the solids into cement

  11. Residual stress concerns in containment analysis

    International Nuclear Information System (INIS)

    Costantini, F.; Kulak, R. F.; Pfeiffer, P. A.

    1997-01-01

    The manufacturing of steel containment vessels starts with the forming of flat plates into curved plates. A steel containment structure is made by welding individual plates together to form the sections that make up the complex shaped vessels. The metal forming and welding process leaves residual stresses in the vessel walls. Generally, the effect of metal forming residual stresses can be reduced or virtually eliminated by thermally stress relieving the vesseL In large containment vessels this may not be practical and thus the residual stresses due to manufacturing may become important. The residual stresses could possibly tiect the response of the vessel to internal pressurization. When the level of residual stresses is significant it will affect the vessel's response, for instance the yielding pressure and possibly the failure pressure. The paper will address the effect of metal forming residual stresses on the response of a generic pressure vessel to internal pressurization. A scoping analysis investigated the effect of residual forming stresses on the response of an internally pressurized vessel. A simple model was developed to gain understanding of the mechanics of the problem. Residual stresses due to the welding process were not considered in this investigation

  12. Electrodialytic remediation of air pollution control residues

    DEFF Research Database (Denmark)

    Jensen, Pernille Erland

    Air pollution control (APC) residue from municipal solid waste incineration (MSWI) consists of the fly ash, and, in dry and semi-dry systems, also the reaction products from the flue gas cleaning process. APC residue is considered a hazardous waste due to its high alkalinity, high content of salt...

  13. Residuals and the Residual-Based Statistic for Testing Goodness of Fit of Structural Equation Models

    Science.gov (United States)

    Foldnes, Njal; Foss, Tron; Olsson, Ulf Henning

    2012-01-01

    The residuals obtained from fitting a structural equation model are crucial ingredients in obtaining chi-square goodness-of-fit statistics for the model. The authors present a didactic discussion of the residuals, obtaining a geometrical interpretation by recognizing the residuals as the result of oblique projections. This sheds light on the…

  14. Functional validation of Ca2+-binding residues from the crystal structure of the BK ion channel.

    Science.gov (United States)

    Kshatri, Aravind S; Gonzalez-Hernandez, Alberto J; Giraldez, Teresa

    2018-04-01

    BK channels are dually regulated by voltage and Ca 2+ , providing a cellular mechanism to couple electrical and chemical signalling. Intracellular Ca 2+ concentration is sensed by a large cytoplasmic region in the channel known as "gating ring", which is formed by four tandems of regulator of conductance for K + (RCK1 and RCK2) domains. The recent crystal structure of the full-length BK channel from Aplysia californica has provided new information about the residues involved in Ca 2+ coordination at the high-affinity binding sites located in the RCK1 and RCK2 domains, as well as their cooperativity. Some of these residues have not been previously studied in the human BK channel. In this work we have investigated, through site directed mutagenesis and electrophysiology, the effects of these residues on channel activation by voltage and Ca 2+ . Our results demonstrate that the side chains of two non-conserved residues proposed to coordinate Ca 2+ in the A. californica structure (G523 and E591) have no apparent functional role in the human BK Ca 2+ sensing mechanism. Consistent with the crystal structure, our data indicate that in the human channel the conserved residue R514 participates in Ca 2+ coordination in the RCK1 binding site. Additionally, this study provides functional evidence indicating that R514 also interacts with residues E902 and Y904 connected to the Ca 2+ binding site in RCK2. Interestingly, it has been proposed that this interaction may constitute a structural correlate underlying the cooperative interactions between the two high-affinity Ca 2+ binding sites regulating the Ca 2+ dependent gating of the BK channel. This article is part of a Special Issue entitled: Beyond the Structure-Function Horizon of Membrane Proteins edited by Ute Hellmich, Rupak Doshi and Benjamin McIlwain. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. LigandRFs: random forest ensemble to identify ligand-binding residues from sequence information alone

    KAUST Repository

    Chen, Peng

    2014-12-03

    Background Protein-ligand binding is important for some proteins to perform their functions. Protein-ligand binding sites are the residues of proteins that physically bind to ligands. Despite of the recent advances in computational prediction for protein-ligand binding sites, the state-of-the-art methods search for similar, known structures of the query and predict the binding sites based on the solved structures. However, such structural information is not commonly available. Results In this paper, we propose a sequence-based approach to identify protein-ligand binding residues. We propose a combination technique to reduce the effects of different sliding residue windows in the process of encoding input feature vectors. Moreover, due to the highly imbalanced samples between the ligand-binding sites and non ligand-binding sites, we construct several balanced data sets, for each of which a random forest (RF)-based classifier is trained. The ensemble of these RF classifiers forms a sequence-based protein-ligand binding site predictor. Conclusions Experimental results on CASP9 and CASP8 data sets demonstrate that our method compares favorably with the state-of-the-art protein-ligand binding site prediction methods.

  16. Recipe for residual oil saturation determination

    Energy Technology Data Exchange (ETDEWEB)

    Guillory, A.J.; Kidwell, C.M.

    1979-01-01

    In 1978, Shell Oil Co., in conjunction with the US Department of Energy, conducted a residual oil saturation study in a deep, hot high-pressured Gulf Coast Reservoir. The work was conducted prior to initiation of CO/sub 2/ tertiary recovery pilot. Many problems had to be resolved prior to and during the residual oil saturation determination. The problems confronted are outlined such that the procedure can be used much like a cookbook in designing future studies in similar reservoirs. Primary discussion centers around planning and results of a log-inject-log operation used as a prime method to determine the residual oil saturation. Several independent methods were used to calculate the residual oil saturation in the subject well in an interval between 12,910 ft (3935 m) and 12,020 ft (3938 m). In general, these numbers were in good agreement and indicated a residual oil saturation between 22% and 24%. 10 references.

  17. Harvesting and handling agricultural residues for energy

    Energy Technology Data Exchange (ETDEWEB)

    Jenkins, B.M.; Summer, H.R.

    1986-05-01

    Significant progress in understanding the needs for design of agricultural residue collection and handling systems has been made but additional research is required. Recommendations are made for research to (a) integrate residue collection and handling systems into general agricultural practices through the development of multi-use equipment and total harvest systems; (b) improve methods for routine evaluation of agricultural residue resources, possibly through remote sensing and image processing; (c) analyze biomass properties to obtain detailed data relevant to engineering design and analysis; (d) evaluate long-term environmental, social, and agronomic impacts of residue collection; (e) develop improved equipment with higher capacities to reduce residue collection and handling costs, with emphasis on optimal design of complete systems including collection, transportation, processing, storage, and utilization; and (f) produce standard forms of biomass fuels or products to enhance material handling and expand biomass markets through improved reliability and automatic control of biomass conversion and other utilization systems. 118 references.

  18. Computational prediction of protein hot spot residues.

    Science.gov (United States)

    Morrow, John Kenneth; Zhang, Shuxing

    2012-01-01

    Most biological processes involve multiple proteins interacting with each other. It has been recently discovered that certain residues in these protein-protein interactions, which are called hot spots, contribute more significantly to binding affinity than others. Hot spot residues have unique and diverse energetic properties that make them challenging yet important targets in the modulation of protein-protein complexes. Design of therapeutic agents that interact with hot spot residues has proven to be a valid methodology in disrupting unwanted protein-protein interactions. Using biological methods to determine which residues are hot spots can be costly and time consuming. Recent advances in computational approaches to predict hot spots have incorporated a myriad of features, and have shown increasing predictive successes. Here we review the state of knowledge around protein-protein interactions, hot spots, and give an overview of multiple in silico prediction techniques of hot spot residues.

  19. Computational Prediction of Hot Spot Residues

    Science.gov (United States)

    Morrow, John Kenneth; Zhang, Shuxing

    2013-01-01

    Most biological processes involve multiple proteins interacting with each other. It has been recently discovered that certain residues in these protein-protein interactions, which are called hot spots, contribute more significantly to binding affinity than others. Hot spot residues have unique and diverse energetic properties that make them challenging yet important targets in the modulation of protein-protein complexes. Design of therapeutic agents that interact with hot spot residues has proven to be a valid methodology in disrupting unwanted protein-protein interactions. Using biological methods to determine which residues are hot spots can be costly and time consuming. Recent advances in computational approaches to predict hot spots have incorporated a myriad of features, and have shown increasing predictive successes. Here we review the state of knowledge around protein-protein interactions, hot spots, and give an overview of multiple in silico prediction techniques of hot spot residues. PMID:22316154

  20. Co-benefits of utilizing logging residues for bioenergy production: The case for East Texas, USA

    International Nuclear Information System (INIS)

    Gan, Jianbang; Smith, C.T.

    2007-01-01

    This study evaluated the co-benefits associated with the utilization of logging residues for electricity production in East Texas, USA. The benefits evaluated included the value of CO 2 emissions displaced due to substituting logging residues for coal in power generation, reductions in site preparation costs during forest regeneration, and creation of jobs and income in local communities. Based on the 2004 Forest Inventory Analysis data and a 70% biomass recovery rate, annual recoverable logging residues in East Texas were estimated at 1.3 Mt (dry). These residues, if used for electricity production, would displace about 2.44 Mt of CO 2 , valued at some 9M$ at the current CO 2 price traded at the Chicago Climate Exchange (accounting for about 2% of the stumpage value). Removing logging residues would also save $200-250 ha -1 in site preparation costs. In addition, input-output modeling revealed that logging residue procurement and electricity generation together would have a stronger ripple effect on employment than on output or value-added, with about 1340 new jobs created and 215M$ in value-added generated annually. These results offer new insights into the cost-competitiveness of forest biomass and bioenergy production. (author)

  1. Identification of key residues for protein conformational transition using elastic network model.

    Science.gov (United States)

    Su, Ji Guo; Xu, Xian Jin; Li, Chun Hua; Chen, Wei Zu; Wang, Cun Xin

    2011-11-07

    Proteins usually undergo conformational transitions between structurally disparate states to fulfill their functions. The large-scale allosteric conformational transitions are believed to involve some key residues that mediate the conformational movements between different regions of the protein. In the present work, a thermodynamic method based on the elastic network model is proposed to predict the key residues involved in protein conformational transitions. In our method, the key functional sites are identified as the residues whose perturbations largely influence the free energy difference between the protein states before and after transition. Two proteins, nucleotide binding domain of the heat shock protein 70 and human/rat DNA polymerase β, are used as case studies to identify the critical residues responsible for their open-closed conformational transitions. The results show that the functionally important residues mainly locate at the following regions for these two proteins: (1) the bridging point at the interface between the subdomains that control the opening and closure of the binding cleft; (2) the hinge region between different subdomains, which mediates the cooperative motions between the corresponding subdomains; and (3) the substrate binding sites. The similarity in the positions of the key residues for these two proteins may indicate a common mechanism in their conformational transitions.

  2. Salmon Site Remedial Investigation Report

    International Nuclear Information System (INIS)

    1999-01-01

    This Salmon Site Remedial Investigation Report provides the results of activities initiated by the U.S. Department of Energy (DOE) to determine if contamination at the Salmon Site poses a current or future risk to human health and the environment. These results were used to develop and evaluate a range of risk-based remedial alternatives. Located in Lamar County, Mississippi, the Salmon Site was used by the U.S. Atomic Energy Commission (predecessor to the DOE) between 1964 and 1970 for two nuclear and two gas explosions conducted deep underground in a salt dome. The testing resulted in the release of radionuclides into the salt dome. During reentry drilling and other site activities, liquid and solid wastes containing radioactivity were generated resulting in surface soil and groundwater contamination. Most of the waste and contaminated soil and water were disposed of in 1993 during site restoration either in the cavities left by the tests or in an injection well. Other radioactive wastes were transported to the Nevada Test Site for disposal. Nonradioactive wastes were disposed of in pits at the site and capped with clean soil and graded. The preliminary investigation showed residual contamination in the Surface Ground Zero mud pits below the water table. Remedial investigations results concluded the contaminant concentrations detected present no significant risk to existing and/or future land users, if surface institutional controls and subsurface restrictions are maintained. Recent sampling results determined no significant contamination in the surface or shallow subsurface. The test cavity resulting from the experiments is contaminated and cannot be economically remediated with existing technologies. The ecological sampling did not detect biological uptake of contaminants in the plants or animals sampled. Based on the current use of the Salmon Site, the following remedial actions were identified to protect both human health and the environment: (1) the

  3. Residual stresses in zircaloy welds

    International Nuclear Information System (INIS)

    Santisteban, J. R.; Fernandez, L; Vizcaino, P.; Banchik, A.D.; Samper, R; Martinez, R. L; Almer, J; Motta, A.T.; Colas, K.B; Kerr, M.; Daymond, M.R

    2009-01-01

    Welds in Zirconium-based alloys are susceptible to hydrogen embrittlement, as H enters the material due to dissociation of water. The yield strain for hydride cracking has a complex dependence on H concentration, stress state and texture. The large thermal gradients produced by the applied heat; drastically changes the texture of the material in the heat affected zone, enhancing the susceptibility to delayed hydride cracking. Normally hydrides tend to form as platelets that are parallel to the normal direction, but when welding plates, hydride platelets may form on cooling with their planes parallel to the weld and through the thickness of the plates. If, in addition to this there are significant tensile stresses, the susceptibility of the heat affected zone to delayed hydride cracking will be increased. Here we have measured the macroscopic and microscopic residual stressed that appear after PLASMA welding of two 6mm thick Zircaloy-4 plates. The measurements were based on neutron and synchrotron diffraction experiments performed at the Isis Facility, UK, and at Advanced Photon Source, USA, respectively. The experiments allowed assessing the effect of a post-weld heat treatment consisting of a steady increase in temperature from room temperature to 450oC over a period of 4.5 hours; followed by cooling with an equivalent cooling rate. Peak tensile stresses of (175± 10) MPa along the longitudinal direction were found in the as-welded specimen, which were moderately reduced to (150±10) MPa after the heat-treatment. The parent material showed intergranular stresses of (56±4) MPa, which disappeared on entering the heat-affected zone. In-situ experiments during themal cyclong of the material showed that these intergranular stresses result from the anisotropy of the thermal expansion coefficient of the hexagonal crystal lattice. [es

  4. Radiological survey of the radioactive sands and residues at Lowman, Idaho

    International Nuclear Information System (INIS)

    Haywood, F.F.; Burden, J.E.; Ellis, B.S.; Loy, E.T.; Shinpaugh, W.H.

    1980-08-01

    No uranium ore milling was performed at the Lowman site, which is located approximately 0.8 km northeast of the town of Lowman, Idaho. Nevertheless, approximately 80,000 metric tons of radioactive sands and residues from upgrading of heavy minerals by physical processing methods remain on the site grounds. Measurements of external gamma radiation 1 m above the surface showed exposure rates up to 2.4 mR/hr on site, but the exposure rate off site quickly dropped to the background level in all directions. Analysis of surface soil and sediment samples for 226 Ra and 232 Th indicated a limited spread of radioactive material

  5. LIBRA: LIgand Binding site Recognition Application.

    Science.gov (United States)

    Hung, Le Viet; Caprari, Silvia; Bizai, Massimiliano; Toti, Daniele; Polticelli, Fabio

    2015-12-15

    In recent years, structural genomics and ab initio molecular modeling activities are leading to the availability of a large number of structural models of proteins whose biochemical function is not known. The aim of this study was the development of a novel software tool that, given a protein's structural model, predicts the presence and identity of active sites and/or ligand binding sites. The algorithm implemented by ligand binding site recognition application (LIBRA) is based on a graph theory approach to find the largest subset of similar residues between an input protein and a collection of known functional sites. The algorithm makes use of two predefined databases for active sites and ligand binding sites, respectively, derived from the Catalytic Site Atlas and the Protein Data Bank. Tests indicate that LIBRA is able to identify the correct binding/active site in 90% of the cases analyzed, 90% of which feature the identified site as ranking first. As far as ligand binding site recognition is concerned, LIBRA outperforms other structure-based ligand binding sites detection tools with which it has been compared. The application, developed in Java SE 7 with a Swing GUI embedding a JMol applet, can be run on any OS equipped with a suitable Java Virtual Machine (JVM), and is available at the following URL: http://www.computationalbiology.it/software/LIBRAv1.zip. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  6. CHARACTERIZATION OF AGROINDUSTRIAL RESIDUES WITH A VIEW TO ITS EXPLOITATION

    Directory of Open Access Journals (Sweden)

    Emir Cabrera Rodríguez

    2016-10-01

    Full Text Available Agroindustrial residues are considered one of the most important renewable resources to obtain products of economic and social interest. This study assesses the potential use of coconut husk, sugar cane straw and sugar cane bagasse marrow through their chemical characterization. For this, the total solid contents, the ash, lignin, holocellulose and extractive contents are determined, as well as the zero charge Ph and the total acid and basic groups of materials. Results show a large lignin percentage in coconut husk composition (42.3 ± 1.03%, which is significantly higher than that of the other residues. That is why this material is a promising feedstock to obtain high-added value chemical products like phenolic compounds, which can replace those derived from crude oil. Straw and marrow have high holocellulose percentages of 61.1 ± 1.0% and 60.3 ± 1.3%, respectively, deserving attention as feedstock for biofuels production and chemical compound synthesis. On the other hand, all residues have higher number of acid sites with a predominance of carboxyl and hydroxyl groups according to infrared spectroscopy (FTIR. Therefore, all materials display potentialities for heavy metals and dissolving colorants biosorption.

  7. Chemical modelling of pore water composition from PFBC residues

    International Nuclear Information System (INIS)

    Karlsson, L.G.

    1991-01-01

    The concentration of trace elements varies depending on the source of the coal and also due to the combustion process used. Mercury is one important element among the trace elements in the coal residues, generally recognised as potentially harmful to the biological system. To predict the pore water concentrations of mercury and other important constituents leached from coal combustion residues disposal sites, mechanistic data on chemical reactions are required. The present study is an application of a basially thermodynamical approach using the geochemical code EQ3NR. The presence of discrete solid phases that control the aqueous concentrations of major elements such as aluminium, calcium and silicon are identified. Solid phases are modelled in equilibrium with a hypothetical pore water at a pH range of 7-11. In this study the thermodynamic database of EQ3NR has been complemented with data for cadmium, mercury and lead taken from the OECD/NEA Thermodynamic Database and from a compilation made by Lindsay. Possible solubility limiting phases for the important trace elements arsenic, cadmium, chromium, copper, mercury, nickel and lead have been identified. Concentrations of these trace elements as a function of pH in the hypothetical pore water were calculated using mechanistic thermodynamial data. The thermodynamical approach in this study seems justified because most solid residues that are either present or expected to form during weathering have relatively fast precipitation/dissolution kinetics. (21 refs., 18 figs., 5 tabs.)

  8. Firework displays as sources of particles similar to gunshot residue.

    Science.gov (United States)

    Grima, Matthew; Butler, Mark; Hanson, Robert; Mohameden, Ahmed

    2012-03-01

    In light of past research being targeted to find specific particles which may be similar to gunshot residue (GSR), this project was formulated to detect any possible particulate by random particle fallout onto substrates at firework displays and to assess the impact this may have on GSR evidence. Firework residue was collected at a display site, from amongst spectators as well as from the author's hair 90min after the display. SEM-EDX analysis has detected such particulate in all three scenarios, with the firework particle population at large providing a solid ground for discrimination from GSR. Wind dispersal was found to decrease the particle population and subsequently, the latter's discriminatory power. Some particles, if treated individually were found to be indistinguishable from GSR. Findings also include residues which may mimic strontium based GSR as well as GSR which may be mixed with that from previous firings. The continuous changes made to primer and propellant compositions by manufacturers also call for greater consideration when classifying particles as originating from pyrotechnic devices. Furthermore, authorities such as police forces should be made more aware about the incidence of such particle transfer in firework related periods. Copyright © 2011 Forensic Science Society. Published by Elsevier Ireland Ltd. All rights reserved.

  9. Applied exposure modeling for residual radioactivity and release criteria

    International Nuclear Information System (INIS)

    Lee, D.W.

    1989-01-01

    The protection of public health and the environment from the release of materials with residual radioactivity for recycle or disposal as wastes without radioactive contents of concern presents a formidable challenge. Existing regulatory criteria are based on technical judgment concerning detectability and simple modeling. Recently, exposure modeling methodologies have been developed to provide a more consistent level of health protection. Release criteria derived from the application of exposure modeling methodologies share the same basic elements of analysis but are developed to serve a variety of purposes. Models for the support of regulations for all applications rely on conservative interpretations of generalized conditions while models developed to show compliance incorporate specific conditions not likely to be duplicated at other sites. Research models represent yet another type of modeling which strives to simulate the actual behavior of released material. In spite of these differing purposes, exposure modeling permits the application of sound and reasoned principles of radiation protection to the release of materials with residual levels of radioactivity. Examples of the similarities and differences of these models are presented and an application to the disposal of materials with residual levels of uranium contamination is discussed. 5 refs., 2 tabs

  10. Residual concentrations of micropollutants in benthic mussels in the coastal areas of Bohai Sea, North China

    International Nuclear Information System (INIS)

    Liu Wenxin; Chen Jianglin; Lin Xiumei; Fan Yongsheng; Tao Shu

    2007-01-01

    Studies of heavy metals and organic pollutants in different benthic mussel species from Bohai Sea show that concentrations of Cd in mussels commonly exceed national biological quality standards. In addition, a site located in Laizhou Bay exhibits higher average concentrations of As, Hg and Pb with respect to the other sites. Residual levels of petroleum hydrocarbons at several sites in Liaodong Bay also exceed quality guidelines. Contents and compositional characteristics of DDT and its metabolites in mussels suggest the probability of recent inputs and potential ecological risks to the local benthic environment. - Residual Cd contents of mussels at many sites in Bohai Sea exceed national marine biological quality standard and compositions of DDT metabolites indicate recent inputs and potential risk

  11. Location of 3-hydroxyproline residues in collagen types I, II, III, and V/XI implies a role in fibril supramolecular assembly.

    Science.gov (United States)

    Weis, Mary Ann; Hudson, David M; Kim, Lammy; Scott, Melissa; Wu, Jiann-Jiu; Eyre, David R

    2010-01-22

    Collagen triple helices are stabilized by 4-hydroxyproline residues. No function is known for the much less common 3-hydroxyproline (3Hyp), although genetic defects inhibiting its formation cause recessive osteogenesis imperfecta. To help understand the pathogenesis, we used mass spectrometry to identify the sites and local sequence motifs of 3Hyp residues in fibril-forming collagens from normal human and bovine tissues. The results confirm a single, essentially fully occupied 3Hyp site (A1) at Pro(986) in A-clade chains alpha1(I), alpha1(II), and alpha2(V). Two partially modified sites (A2 and A3) were found at Pro(944) in alpha1(II) and alpha2(V) and Pro(707) in alpha2(I) and alpha2(V), which differed from A1 in sequence motif. Significantly, the distance between sites 2 and 3, 237 residues, is close to the collagen D-period (234 residues). A search for additional D-periodic 3Hyp sites revealed a fourth site (A4) at Pro(470) in alpha2(V), 237 residues N-terminal to site 3. In contrast, human and bovine type III collagen contained no 3Hyp at any site, despite a candidate proline residue and recognizable A1 sequence motif. A conserved histidine in mammalian alpha1(III) at A1 may have prevented 3-hydroxylation because this site in chicken type III was fully hydroxylated, and tyrosine replaced histidine. All three B-clade type V/XI collagen chains revealed the same three sites of 3Hyp but at different loci and sequence contexts from those in A-clade collagen chains. Two of these B-clade sites were spaced apart by 231 residues. From these and other observations we propose a fundamental role for 3Hyp residues in the ordered self-assembly of collagen supramolecular structures.

  12. Quantification of residual stress from photonic signatures of fused silica

    International Nuclear Information System (INIS)

    Cramer, K. Elliott; Yost, William T.; Hayward, Maurice

    2014-01-01

    A commercially available grey-field polariscope (GFP) instrument for photoelastic examination is used to assess impact damage inflicted upon the outer-most pane of Space Shuttle windows made from fused silica. A method and apparatus for calibration of the stress-optic coefficient using four-point bending is discussed. The results are validated on known material (acrylic) and are found to agree with literature values to within 6%. The calibration procedure is then applied to fused-silica specimens and the stress-optic coefficient is determined to be 2.43 ± 0.54 × 10 −12 Pa −1 . Fused silica specimens containing impacts artificially made at NASA’s Hypervelocity Impact Technology Facility (HIT-F), to simulate damage typical during space flight, are examined. The damage sites are cored from fused silica window carcasses and examined with the GFP. The calibrated GFP measurements of residual stress patterns surrounding the damage sites are presented

  13. Effect of texture and grain size on the residual stress of nanocrystalline thin films

    Science.gov (United States)

    Cao, Lei; Sengupta, Arkaprabha; Pantuso, Daniel; Koslowski, Marisol

    2017-10-01

    Residual stresses develop in thin film interconnects mainly as a result of deposition conditions and multiple thermal loading cycles during the manufacturing flow. Understanding the relation between the distribution of residual stress and the interconnect microstructure is of key importance to manage the nucleation and growth of defects that can lead to failure under reliability testing and use conditions. Dislocation dynamics simulations are performed in nanocrystalline copper subjected to cyclic loading to quantify the distribution of residual stresses as a function of grain misorientation and grain size distribution. The outcomes of this work help to evaluate the effect of microstructure in thin films failure by identifying potential voiding sites. Furthermore, the simulations show how dislocation structures are influenced by texture and grain size distribution that affect the residual stress. For example, when dislocation loops reach the opposite grain boundary during loading, these dislocations remain locked during unloading.

  14. Positive-Unlabeled Learning for Pupylation Sites Prediction

    Directory of Open Access Journals (Sweden)

    Ming Jiang

    2016-01-01

    Full Text Available Pupylation plays a key role in regulating various protein functions as a crucial posttranslational modification of prokaryotes. In order to understand the molecular mechanism of pupylation, it is important to identify pupylation substrates and sites accurately. Several computational methods have been developed to identify pupylation sites because the traditional experimental methods are time-consuming and labor-sensitive. With the existing computational methods, the experimentally annotated pupylation sites are used as the positive training set and the remaining nonannotated lysine residues as the negative training set to build classifiers to predict new pupylation sites from the unknown proteins. However, the remaining nonannotated lysine residues may contain pupylation sites which have not been experimentally validated yet. Unlike previous methods, in this study, the experimentally annotated pupylation sites were used as the positive training set whereas the remaining nonannotated lysine residues were used as the unlabeled training set. A novel method named PUL-PUP was proposed to predict pupylation sites by using positive-unlabeled learning technique. Our experimental results indicated that PUL-PUP outperforms the other methods significantly for the prediction of pupylation sites. As an application, PUL-PUP was also used to predict the most likely pupylation sites in nonannotated lysine sites.

  15. Predictive hydrogeochemical modelling of bauxite residue sand in field conditions.

    Science.gov (United States)

    Wissmeier, Laurin; Barry, David A; Phillips, Ian R

    2011-07-15

    The suitability of residue sand (the coarse fraction remaining from Bayer's process of bauxite refining) for constructing the surface cover of closed bauxite residue storage areas was investigated. Specifically, its properties as a medium for plant growth are of interest to ensure residue sand can support a sustainable ecosystem following site closure. The geochemical evolution of the residue sand under field conditions, its plant nutrient status and soil moisture retention were studied by integrated modelling of geochemical and hydrological processes. For the parameterization of mineral reactions, amounts and reaction kinetics of the mineral phases natron, calcite, tricalcium aluminate, sodalite, muscovite and analcime were derived from measured acid neutralization curves. The effective exchange capacity for ion adsorption was measured using three independent exchange methods. The geochemical model, which accounts for mineral reactions, cation exchange and activity corrected solution speciation, was formulated in the geochemical modelling framework PHREEQC, and partially validated in a saturated-flow column experiment. For the integration of variably saturated flow with multi-component solute transport in heterogeneous 2D domains, a coupling of PHREEQC with the multi-purpose finite-element solver COMSOL was established. The integrated hydrogeochemical model was applied to predict water availability and quality in a vertical flow lysimeter and a cover design for a storage facility using measured time series of rainfall and evaporation from southwest Western Australia. In both scenarios the sand was fertigated and gypsum-amended. Results show poor long-term retention of fertilizer ions and buffering of the pH around 10 for more than 5 y of leaching. It was concluded that fertigation, gypsum amendment and rainfall leaching alone were insufficient to render the geochemical conditions of residue sand suitable for optimal plant growth within the given timeframe. The

  16. A look at worldwide usage of residual wood for energy

    International Nuclear Information System (INIS)

    Ekstrom, H.; Hall, M.M.

    2007-01-01

    Wood Resources International was established in 1987, offering on-site evaluation services of forest resources and forest industry developments in over 20 countries worldwide. This presentation reviewed residual wood markets in North America and Europe. Wood chip trade and wood pellet markets were also reviewed. It is estimated that more than 50 per cent of the wood harvested worldwide is used for heating and cooking. Although sawmill wood residue has been typically used for particle board manufacturing, the energy sector in North America and Europe is now competing for low cost residuals, including sawdust, shavings and wood chips. With demand for renewable resources increasing, district heating plants have revived an interest in collecting the nearly 35 per cent of biomass left behind after traditional clear cutting. This biomass represents branches, tops and stumps left behind after the roundwood has been removed. In Canada, demand for mill residuals has grown and wood pellet manufacturers have the opportunity to invest in capacity while continuing to produce competitively priced pellets for the European market. It is anticipated that in the next decade, large volumes of beetle-killed wood are going to be available in British Columbia for energy consumption, including wood pellet production. Prices for sawdust have doubled over the past 3 years as a result of increased competition. The biomass supply potential in the United States is 7 times the current consumption. There is an increased interest in bioenergy in California due to the declining lumber sector. As such, the use of forest and agricultural waste is on the rise, along with prices for wood residues. There has also been a large increase in demand for wood biomass in Europe over the past 5 years, resulting in higher costs of all wood fiber sources used for energy. By 2020, Europe has set a target that all energy should come from renewable energy sources, with a minimum of 10 per cent being biofuel for

  17. Residues from waste incineration. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Astrup, T.; Juul Pedersen, A.; Hyks, J.; Frandsen, F.J.

    2009-08-15

    The overall objective of the project was to improve the understanding of the formation and characteristics of residues from waste incineration. This was done focusing on the importance of the waste input and the operational conditions of the furnace. Data and results obtained from the project have been discussed in this report according to the following three overall parts: i) mass flows and element distribution, ii) flue gas/particle partitioning and corrosion/deposition aspects, and iii) residue leaching. This has been done with the intent of structuring the discussion while tacitly acknowledging that these aspects are interrelated and cannot be separated. Overall, it was found that the waste input composition had significant impact of the characteristics of the generated residues. A similar correlation between operational conditions and residue characteristics could not be observed. Consequently, the project recommend that optimization of residue quality should focus on controlling the waste input composition. The project results showed that including specific waste materials (and thereby also excluding the same materials) may have significant effects on the residue composition, residue leaching, aerosol and deposit formation.It is specifically recommended to minimize Cl in the input waste. Based on the project results, it was found that a significant potential for optimization of waste incineration exist. (author)

  18. ENTRIA 2014. Memorandum on the disposal of high-level radioactive residuals

    International Nuclear Information System (INIS)

    Roehlig, Klaus-Juergen; Walther, Clemens; Bach, Friedrich-Wilhelm

    2014-01-01

    The memorandum on the disposal of high-level radioactive residuals covers the following issues: description of the problem: a ''wicked problem'', risks and NIMBY, the site selection law, international boundary conditions; disposal strategy and types of facilities: safety and reversibility, long-term surface storage, deep storage; risk and safety; procedural justice and the site selection process; social innovations and the requirement of long-term institutions; conclusion - central stress fields.

  19. Use of ultrasound in petroleum residue upgradation

    Energy Technology Data Exchange (ETDEWEB)

    Sawarkar, A.N.; Pandit, A.B.; Samant, S.D.; Joshi, J.B. [Mumbai Univ., Mumbai (India). Inst. of Chemical Technology

    2009-06-15

    The importance of bottom-of-the barrel upgrading has increased in the current petroleum refining scenario because of the progressively heavier nature of crude oil. Heavy residues contain large concentrations of metals such as vanadium and nickel which foul catalysts and reduce the potential effect of residue fluidized catalytic cracking. This study showed that the cavitational energy induced by ultrasound be be successfully used to upgrade hydrocarbon mixtures. Conventional processes for the upgrading of residual feedstocks, such as thermal cracking and catalytic cracking, were carried out in the temperature range of 400-520 degrees C. Experiments were performed on 2 vacuum residues, Arabian mix vacuum residue (AMVR) and Bombay high vacuum residue (BHVR) and 1 Haldia asphalt (HA). These were subjected to acoustic cavitation for different reaction times from 15 to 120 minutes at ambient temperature and pressure. Two acoustic cavitation devices were compared, namely the ultrasonic bath and ultrasonic horn. In particular, this study compared the ability of these 2 devices to upgrade the petroleum residues to lighter, more value-added products. Different surfactants were used to examine the effect of ultrasound on upgrading the residue when emulsified in water. In order to better understand the reaction mechanism, a kinetic model was developed based on the constituents of the residue. The ultrasonic horn was found to be more effective in bringing about the upgrading than ultrasonic bath. The study also showed that the acoustic cavitation of the aqueous emulsified hydrocarbon mixture could reduce the asphaltenes content to a greater extent than the acoustic cavitation of non-emulsified hydrocarbon mixture. 20 refs., 11 tabs., 17 figs.

  20. Crop residue harvest for bioenergy production and its implications on soil functioning and plant growth: A review

    Directory of Open Access Journals (Sweden)

    Maurício Roberto Cherubin

    Full Text Available ABSTRACT: The use of crop residues as a bioenergy feedstock is considered a potential strategy to mitigate greenhouse gas (GHG emissions. However, indiscriminate harvesting of crop residues can induce deleterious effects on soil functioning, plant growth and other ecosystem services. Here, we have summarized the information available in the literature to identify and discuss the main trade-offs and synergisms involved in crop residue management for bioenergy production. The data consistently showed that crop residue harvest and the consequent lower input of organic matter into the soil led to C storage depletions over time, reducing cycling, supply and availability of soil nutrients, directly affecting the soil biota. Although the biota regulates key functions in the soil, crop residue can also cause proliferation of some important agricultural pests. In addition, crop residues act as physical barriers that protect the soil against raindrop impact and temperature variations. Therefore, intensive crop residue harvest can cause soil structure degradation, leading to soil compaction and increased risks of erosion. With regard to GHG emissions, there is no consensus about the potential impact of management of crop residue harvest. In general, residue harvest decreases CO2 and N2O emissions from the decomposition process, but it has no significant effect on CH4 emissions. Plant growth responses to soil and microclimate changes due to crop residue harvest are site and crop specific. Adoption of the best management practices can mitigate the adverse impacts of crop residue harvest. Longterm experiments within strategic production regions are essential to understand and monitor the impact of integrated agricultural systems and propose customized solutions for sustainable crop residue management in each region or landscape. Furthermore, private and public investments/cooperations are necessary for a better understanding of the potential environmental

  1. Quantification of Residual Stress from Photonic Signatures of Fused Silica

    Science.gov (United States)

    Cramer, K. Elliott; Hayward, Maurice; Yost, William E.

    2013-01-01

    A commercially available grey-field polariscope (GFP) instrument for photoelastic examination is used to assess impact damage inflicted upon the outer-most pane of Space Shuttle windows made from fused silica. A method and apparatus for calibration of the stress-optic coefficient using four-point bending is discussed. The results are validated on known material (acrylic) and are found to agree with literature values to within 6%. The calibration procedure is then applied to fused-silica specimens and the stress-optic coefficient is determined to be 2.43 +/- 0.54 x 10(exp -12)/Pa. Fused silica specimens containing impacts artificially made at NASA's Hypervelocity Impact Technology Facility (HIT-F), to simulate damage typical during space flight, are examined. The damage sites are cored from fused silica window carcasses and examined with the GFP. The calibrated GFP measurements of residual stress patterns surrounding the damage sites are presented. Keywords: Glass, fused silica, photoelasticity, residual stress

  2. Residual stress analysis in thick uranium films

    International Nuclear Information System (INIS)

    Hodge, A.M.; Foreman, R.J.; Gallegos, G.F.

    2005-01-01

    Residual stress analysis was performed on thick, 1-25 μm, depleted uranium (DU) films deposited on an Al substrate by magnetron sputtering. Two distinct characterization techniques were used to measure substrate curvature before and after deposition. Stress evaluation was performed using the Benabdi/Roche equation, which is based on beam theory of a bi-layer material. The residual stress evolution was studied as a function of coating thickness and applied negative bias voltage (0, -200, -300 V). The stresses developed were always compressive; however, increasing the coating thickness and applying a bias voltage presented a trend towards more tensile stresses and thus an overall reduction of residual stresses

  3. Residues in food derived from animals

    International Nuclear Information System (INIS)

    Grossklaus, D.

    1989-01-01

    The first chapter presents a survey of fundamentals and methods of the detection and analysis of residues in food derived from animals, also referring to the resulting health hazards to man, and to the relevant legal provisions. The subsequent chapters have been written by experts of the Federal Health Office, each dealing with particular types of residues such as those of veterinary drugs, additives to animal feeds, pesticide residues, and with environmental pollutants and the contamination of animal products with radionuclides. (MG) With 35 figs., 61 tabs [de

  4. Probing binding hot spots at protein-RNA recognition sites.

    Science.gov (United States)

    Barik, Amita; Nithin, Chandran; Karampudi, Naga Bhushana Rao; Mukherjee, Sunandan; Bahadur, Ranjit Prasad

    2016-01-29

    We use evolutionary conservation derived from structure alignment of polypeptide sequences along with structural and physicochemical attributes of protein-RNA interfaces to probe the binding hot spots at protein-RNA recognition sites. We find that the degree of conservation varies across the RNA binding proteins; some evolve rapidly compared to others. Additionally, irrespective of the structural class of the complexes, residues at the RNA binding sites are evolutionary better conserved than those at the solvent exposed surfaces. For recognitions involving duplex RNA, residues interacting with the major groove are better conserved than those interacting with the minor groove. We identify multi-interface residues participating simultaneously in protein-protein and protein-RNA interfaces in complexes where more than one polypeptide is involved in RNA recognition, and show that they are better conserved compared to any other RNA binding residues. We find that the residues at water preservation site are better conserved than those at hydrated or at dehydrated sites. Finally, we develop a Random Forests model using structural and physicochemical attributes for predicting binding hot spots. The model accurately predicts 80% of the instances of experimental ΔΔG values in a particular class, and provides a stepping-stone towards the engineering of protein-RNA recognition sites with desired affinity. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

  5. Report of the Siting Policy Task Force

    International Nuclear Information System (INIS)

    1979-08-01

    In August 1978, the Nuclear Regulatory Commission directed the staff to develop a general policy statement on nuclear power reactor siting. A Task Force was formed for that purpose and has prepared a statement of current NRC policy and practice and has recommended a number of changes to current policy. Recommendations were made to accomplish the following goals: (1) To strengthen siting as a factor in defense in depth by establishing requirements for site approval that are independent of plant design consideration. The present policy of permitting plant design features to compensate for unfavorable site characteristics has resulted in improved designs but has tended to deemphasize site isolation. (2) To take into consideration in siting the risk associated with accidents beyond the design basis (Class 9) by establishing population density and distribution criteria. Plant design improvements have reduced the probability and consequences of design basis accidents, but there remains the residual risk from accidents not considered in the design basis. Although this risk cannot be completely reduced to zero, it can be significantly reduced by selective siting. (3) To require that sites selected will minimize the risk from energy generation. The selected sites should be among the best available in the region where new generating capacity is needed. Siting requirements should be stringent enough to limit the residual risk of reactor operation but not so stringent as to eliminate the nuclear option from large regions of the country. This is because energy generation from any source has its associated risk, with risks from some energy sources being greater than that of the nuclear option

  6. Guidelines for selection and presentation of residue values of pesticides

    NARCIS (Netherlands)

    Velde-Koerts T van der; Hoeven-Arentzen PH van; Ossendorp BC; RIVM-SIR

    2004-01-01

    Pesticide residue assessments are executed to establish legal limits, called Maximum Residue Limits (MRLs). MRLs are derived from the results of these pesticide residue trials, which are performed according to critical Good Agricultural Practice. Only one residue value per residue trial may be

  7. Integrating Residual Stress Analysis of Critical Fastener Holes into USAF Depot Maintenance

    Science.gov (United States)

    2014-11-02

    Residual Stress Final Report Contract# FA9453-12-C-0218 Figure 6 shows a typical optical fractograph. The particle at the crack nucleation site is...with a relative humidity < 40% (air conditioned) and a temperature of approximately 75°F. Frequency for the spectrum was 8 Hz, which was governed

  8. Sources of manganese in the residue from a water treatment plant ...

    African Journals Online (AJOL)

    Disposal of water treatment residue (WTR), the by-product from the production of potable water, has traditionally been to landfill. The shortage of suitable landfill sites has led to the proposal that WTR be applied to land. Such disposal is only possible if the WTR contains no toxic elements that may contaminate soil, water or ...

  9. The hydrogen bonds between Arg423 and Glu472 and other key residues, Asp443, Ser477, and Pro489, are responsible for the formation and a different positioning of TNP-ATP and ATP within the nucleotide-binding site of Na(+)/K(+)-ATPase

    Czech Academy of Sciences Publication Activity Database

    Lánský, Zdeněk; Kubala, Martin; Ettrich, Rüdiger; Kutý, Michal; Plášek, J.; Teisinger, Jan; Schoner, W.; Amler, Evžen

    2004-01-01

    Roč. 43, č. 26 (2004), s. 8303-8311 ISSN 0006-2960 R&D Projects: GA MŠk LN00A141; GA ČR GP206/03/D082; GA ČR GA309/02/1479; GA ČR GD305/03/H148 Institutional research plan: CEZ:AV0Z5011922; CEZ:MSM 113100001; CEZ:MSM 111300002 Keywords : sodium pump * ATP-binding site * TNP-ATP Subject RIV: CE - Biochemistry Impact factor: 4.008, year: 2004

  10. Evaluation and optimization of nutritional and environmental impact of biogas residues; Bewertung und Optimierung der Naehrstoff- und Umweltwirkung von Gaerrueckstaenden aus der Biogasgewinnung

    Energy Technology Data Exchange (ETDEWEB)

    Lichti, Fabian Heribert

    2013-04-29

    On the basis of the dynamic growth of biogas plants in Germany the fertilization with biogas residues has obtained an important role for recirculation of plant nutrients, particularly with regard to nitrogen. In this work the effect of N nutrition with biogas residues was assessed in a 3-year on-field trial conducted at four sites throughout Bavaria. The fertilizing effects were tested by varying rate and time of biogas residues application, using different application techniques and the addition of nitrification inhibitors on several crops. The biogas residues achieved mineral fertilizer equivalents of 30 - 45 %. Overall, the untreated biogas residues showed a slightly increased N efficiency compared to cattle manure, whereas particularly site-dependent differences resulted in large differences in N efficiency of biogas residues.

  11. Contaminated Sites in Iowa

    Data.gov (United States)

    Iowa State University GIS Support and Research Facility — Sites contaminated by hazardous materials or wastes. These sites are those administered by the Contaminated Sites Section of Iowa DNR. Many are sites which are...

  12. Radiation doses from residual radioactivity

    International Nuclear Information System (INIS)

    Okajima, Shunzo; Fujita, Shoichiro; Harley, John H.

    1987-01-01

    requires knowing the location of the person to within about 200 m from the time of the explosion to a few weeks afterwards. This is an effort that might be comparable to the present shielding study for survivors. The sizes of the four exposed groups are relatively small; however, the number has been estimated only for those exposed to fallout in the Nishiyama district of Nagasaki. Okajima listed the population of Nishiyama as about 600 at the time of the bomb. No figures are available for the other three groups. The individual exposures from residual radiation may not be significant compared with the direct radiation at the time of the bomb. On the other hand, individuals with potential exposure from these sources are dubious candidates for inclusion in a cohort that was presumably not exposed. For comparison with organ doses estimated in other parts of this program, the exposure estimates are converted to absorbed dose in tissue. The first conversion of exposure to absorbed dose in air uses the factor rad in air 0.87 x exposure in R. UNSCEAR uses an average combined factor of 0.7 to convert absorbed dose in air to absorbed dose in tissue for the whole body. This factor accounts for the change in material (air to tissue) and for backscatter and the shielding afforded by other tissues of the body. No allowance for shielding by buildings has been included here. The cumulative fallout exposures given above become absorbed doses in tissue of 12 to 24 rad for Nagasaki and 0.6 to 2 rad for Hiroshima. The cumulative exposures from induced radioactivity become absorbed doses in tissue of 18 to 24 rad for Nagasaki and about 50 rad for Hiroshima. (author)

  13. Liquefied Residue of Kenaf Core Wood Produced at Different Phenol-Kenaf Ratio

    International Nuclear Information System (INIS)

    Saiful Bahari Bakarudin; Sarani Zakaria; Chia, C.H.; Jani, S.M.

    2012-01-01

    Liquefactions of kenaf core wood were carried out at different phenol-kenaf (P/ k) ratios. Characterizations of kenaf core wood liquefied residue were carried out to measure the degree of liquefaction. This provides a new approach to understand some fundamental aspects of the liquefaction reaction. Functional groups on the raw kenaf core wood and liquefied residue were examined using Fourier transform infrared spectroscopy (FTIR). The crystallinity index of the kenaf wood liquefied residue, which represents crystallinity changes of the cellulose component after the liquefaction process, was studied using X-ray diffraction (XRD). The surface morphology of the wood residue was observed using scanning electron microscopy (SEM). The thermal behavior of the residues was analyzed using thermogravimetric analysis (TGA). Abroad peak around 3450-3400 cm -1 representing OH stretching in lignin start to disappear as P/K ratio increases. The results showed that the higher the P/K ratio the greater the liquefaction of the lignin component in the kenaf core wood. The crystallinity index (CrI) on the kenaf liquefied residues increased with the increase in P/K ratio. SEM images showed that the small fragments attached on the liquefied kenaf residue surface were gradually removed as the P/K ratio was increased from 1.5/ 1.0 to 2.5/ 1.0, which is mainly attributed to the greater chemical penetration toward reactive site of the kenaf fibres. Residue content decreased as the P/K ratio increased from 1.5/ 1.0 to 2.5/ 1.0. TGA results showed the increase of heat resistance in the residue as the P/K ratio was increased. (author)

  14. Cyolane residues in milk of lactating goats

    International Nuclear Information System (INIS)

    Zayed, S.M.A.D.; Osman, A.; Fakhr, I.M.I.

    1981-01-01

    Consecutive feeding of lactating goats with 14 C-alkyl labelled cyolane for 5 days at dietary levels 8 and 16 ppm resulted in the appearance of measurable insecticide residues in milk (0.02-0.04 mg/kg). The residue levels were markedly reduced after a withdrawal period of 7 days. Analysis of urine and milk residues showed the presence of similar metabolites in addition to the parent compound. The major part of the residue consisted of mono-, diethyl phosphate and 2 hydrophilic unknown metabolites. The erythrocyte cholinesterase activity was reduced to about 50% after 24 hours whereas the plasma enzyme was only slightly affected. The animals remained symptom-free during the experimental period. (author)

  15. Surgical treatment for residual or recurrent strabismus

    Directory of Open Access Journals (Sweden)

    Tao Wang

    2014-12-01

    Full Text Available Although the surgical treatment is a relatively effective and predictable method for correcting residual or recurrent strabismus, such as posterior fixation sutures, medial rectus marginal myotomy, unilateral or bilateral rectus re-recession and resection, unilateral lateral rectus recession and adjustable suture, no standard protocol is established for the surgical style. Different surgical approaches have been recommended for correcting residual or recurrent strabismus. The choice of the surgical procedure depends on the former operation pattern and the surgical dosages applied on the patients, residual or recurrent angle of deviation and the operator''s preference and experience. This review attempts to outline recent publications and current opinion in the management of residual or recurrent esotropia and exotropia.

  16. Recovery of transuranics from process residues

    International Nuclear Information System (INIS)

    Gray, J.H.; Gray, L.W.

    1987-01-01

    Process residues are generated at both the Rocky Flats Plant (RFP) and the Savannah River Plant (SRP) during aqueous chemical and pyrochemical operations. Frequently, process operations will result in either impure products or produce residues sufficiently contaminated with transuranics to be nondiscardable as waste. Purification and recovery flowsheets for process residues have been developed to generate solutions compatible with subsequent Purex operations and either solid or liquid waste suitable for disposal. The ''scrub alloy'' and the ''anode heel alloy'' are examples of materials generated at RFP which have been processed at SRP using the developed recovery flowsheets. Examples of process residues being generated at SRP for which flowsheets are under development include LECO crucibles and alpha-contaminated hydraulic oil

  17. U.S. Isostatic Residual Gravity Grid

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — isores.bin - standard grid containing isostatic residual gravity map for U.S. Grid interval = 4 km. Projection is Albers (central meridian = 96 degrees West; base...

  18. Management of stormwater facility maintenance residuals

    Science.gov (United States)

    1998-06-01

    Current research on stormwater maintenance residuals has revealed that the source and nature of these materials is extremely variable, that regulation can be ambiguous, and handling can be costly and difficult. From a regulatory perspective, data ind...

  19. Residual stresses in Inconel 718 engine disks

    Directory of Open Access Journals (Sweden)

    Dahan Yoann

    2014-01-01

    Full Text Available Aubert&Duval has developed a methodology to establish a residual stress model for Inconel 718 engine discs. To validate the thermal, mechanical and metallurgical parts of the model, trials on lab specimens with specific geometry were carried out. These trials allow a better understanding of the residual stress distribution and evolution during different processes (quenching, ageing, machining. A comparison between experimental and numerical results reveals the residual stresses model accuracy. Aubert&Duval has also developed a mechanical properties prediction model. Coupled with the residual stress prediction model, Aubert&Duval can now propose improvements to the process of manufacturing in Inconel 718 engine disks. This model enables Aubert&Duval customers and subcontractors to anticipate distortions issues during machining. It could also be usedt to optimise the engine disk life.

  20. [Development of residual voltage testing equipment].

    Science.gov (United States)

    Zeng, Xiaohui; Wu, Mingjun; Cao, Li; He, Jinyi; Deng, Zhensheng

    2014-07-01

    For the existing measurement methods of residual voltage which can't turn the power off at peak voltage exactly and simultaneously display waveforms, a new residual voltage detection method is put forward in this paper. First, the zero point of the power supply is detected with zero cross detection circuit and is inputted to a single-chip microcomputer in the form of pulse signal. Secend, when the zero point delays to the peak voltage, the single-chip microcomputer sends control signal to power off the relay. At last, the waveform of the residual voltage is displayed on a principal computer or oscilloscope. The experimental results show that the device designed in this paper can turn the power off at peak voltage and is able to accurately display the voltage waveform immediately after power off and the standard deviation of the residual voltage is less than 0.2 V at exactly one second and later.


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