WorldWideScience

Sample records for site-specific labeling method

  1. Electron paramagnetic resonance spin label titration: a novel method to investigate random and site-specific immobilization of enzymes onto polymeric membranes with different properties

    International Nuclear Information System (INIS)

    Butterfield, D. Allan; Colvin, Joshua; Liu Jiangling; Wang Jianquan; Bachas, Leonidas; Bhattacharrya, Dibakar

    2002-01-01

    The immobilization of biological molecules onto polymeric membranes to produce biofunctional membranes is used for selective catalysis, separation, analysis, and artificial organs. Normally, random immobilization of enzymes onto polymeric membranes leads to dramatic reduction in activity due to chemical reactions involved in enzyme immobilization, multiple-point binding, etc., and the extent of activity reduction is a function of membrane hydrophilicity (e.g. activity in cellulosic membrane >> polysulfone membrane). We have used molecular biology to effect site-specific immobilization of enzymes in a manner that orients the active site away from the polymeric membrane surface, thus resulting in higher enzyme activity that approaches that in solution and in increased stability of the enzyme relative to the enzyme in solution. A prediction of this site-specific method of enzyme immobilization, which in this study with subtilisin and organophosphorus hydrolase consists of a fusion tag genetically added to these enzymes and subsequent immobilization via the anti-tag antibody and membrane-bound protein A, is that the active site conformation will more closely resemble that of the enzyme in solution than is the case for random immobilization. This hypothesis was confirmed using a new electron paramagnetic resonance (EPR) spin label active site titration method that determines the amount of spin label bound to the active site of the immobilized enzyme. This value nearly perfectly matched the enzyme activity, and the results suggested: (a) a spectroscopic method for measuring activity and thus the extent of active enzyme immobilization in membrane, which may have advantages in cases where optical methods can not be used due to light scattering interference; (b) higher spin label incorporation (and hence activity) in enzymes that had been site-specifically immobilized versus random immobilization; (c) higher spin label incorporation in enzymes immobilized onto hydrophilic

  2. Site-Specific Biomolecule Labeling with Gold Clusters

    Science.gov (United States)

    Ackerson, Christopher J.; Powell, Richard D.; Hainfeld, James F.

    2013-01-01

    Site-specific labeling of biomolecules in vitro with gold clusters can enhance the information content of electron cryomicroscopy experiments. This chapter provides a practical overview of well-established techniques for forming biomolecule/gold cluster conjugates. Three bioconjugation chemistries are covered: Linker-mediated bioconjugation, direct gold–biomolecule bonding, and coordination-mediated bonding of nickel(II) nitrilotriacetic acid (NTA)-derivatized gold clusters to polyhistidine (His)-tagged proteins. PMID:20887859

  3. Site-Specific Biomolecule Labeling with Gold Clusters

    OpenAIRE

    Ackerson, Christopher J.; Powell, Richard D.; Hainfeld, James F.

    2010-01-01

    Site-specific labeling of biomolecules in vitro with gold clusters can enhance the information content of electron cryomicroscopy experiments. This chapter provides a practical overview of well-established techniques for forming biomolecule/gold cluster conjugates. Three bioconjugation chemistries are covered: Linker-mediated bioconjugation, direct gold–biomolecule bonding, and coordination-mediated bonding of nickel(II) nitrilotriacetic acid (NTA)-derivatized gold clusters to polyhistidine (...

  4. Site-Specific Bioorthogonal Labeling for Fluorescence Imaging of Intracellular Proteins in Living Cells.

    Science.gov (United States)

    Peng, Tao; Hang, Howard C

    2016-11-02

    Over the past years, fluorescent proteins (e.g., green fluorescent proteins) have been widely utilized to visualize recombinant protein expression and localization in live cells. Although powerful, fluorescent protein tags are limited by their relatively large sizes and potential perturbation to protein function. Alternatively, site-specific labeling of proteins with small-molecule organic fluorophores using bioorthogonal chemistry may provide a more precise and less perturbing method. This approach involves site-specific incorporation of unnatural amino acids (UAAs) into proteins via genetic code expansion, followed by bioorthogonal chemical labeling with small organic fluorophores in living cells. While this approach has been used to label extracellular proteins for live cell imaging studies, site-specific bioorthogonal labeling and fluorescence imaging of intracellular proteins in live cells is still challenging. Herein, we systematically evaluate site-specific incorporation of diastereomerically pure bioorthogonal UAAs bearing stained alkynes or alkenes into intracellular proteins for inverse-electron-demand Diels-Alder cycloaddition reactions with tetrazine-functionalized fluorophores for live cell labeling and imaging in mammalian cells. Our studies show that site-specific incorporation of axial diastereomer of trans-cyclooct-2-ene-lysine robustly affords highly efficient and specific bioorthogonal labeling with monosubstituted tetrazine fluorophores in live mammalian cells, which enabled us to image the intracellular localization and real-time dynamic trafficking of IFITM3, a small membrane-associated protein with only 137 amino acids, for the first time. Our optimized UAA incorporation and bioorthogonal labeling conditions also enabled efficient site-specific fluorescence labeling of other intracellular proteins for live cell imaging studies in mammalian cells.

  5. Sensing site-specific structural characteristics and chirality using vibrational circular dichroism of isotope labeled peptides.

    Science.gov (United States)

    Keiderling, Timothy A

    2017-12-01

    Isotope labeling has a long history in chemistry as a tool for probing structure, offering enhanced sensitivity, or enabling site selection with a wide range of spectroscopic tools. Chirality sensitive methods such as electronic circular dichroism are global structural tools and have intrinsically low resolution. Consequently, they are generally insensitive to modifications to enhance site selectivity. The use of isotope labeling to modify vibrational spectra with unique resolvable frequency shifts can provide useful site-specific sensitivity, and these methods have been recently more widely expanded in biopolymer studies. While the spectral shifts resulting from changes in isotopic mass can provide resolution of modes from specific parts of the molecule and can allow detection of local change in structure with perturbation, these shifts alone do not directly indicate structure or chirality. With vibrational circular dichroism (VCD), the shifted bands and their resultant sign patterns can be used to indicate local conformations in labeled biopolymers, particularly if multiple labels are used and if their coupling is theoretically modeled. This mini-review discusses selected examples of the use of labeling specific amides in peptides to develop local structural insight with VCD spectra. © 2017 Wiley Periodicals, Inc.

  6. Site-specific fluorescent labeling of nascent proteins on the translating ribosome.

    Science.gov (United States)

    Saraogi, Ishu; Zhang, Dawei; Chandrasekaran, Sandhya; Shan, Shu-ou

    2011-09-28

    As newly synthesized proteins emerge from the ribosome, they interact with a variety of cotranslational cellular machineries that facilitate their proper folding, maturation, and localization. These interactions are essential for proper function of the cell, and the ability to study these events is crucial to understanding cellular protein biogenesis. To this end, we have developed a highly efficient method to generate ribosome-nascent chain complexes (RNCs) site-specifically labeled with a fluorescent dye on the nascent polypeptide. The fluorescent RNC provides real-time, quantitative information on its cotranslational interaction with the signal recognition particle and will be a valuable tool in elucidating the role of the translating ribosome in numerous biochemical pathways.

  7. 40 CFR 170.232 - Knowledge of labeling and site-specific information.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Knowledge of labeling and site-specific information. 170.232 Section 170.232 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS WORKER PROTECTION STANDARD Standard for Pesticide Handlers § 170.232 Knowledge...

  8. Site-specifically {sup 89}Zr-labeled monoclonal antibodies for ImmunoPET

    Energy Technology Data Exchange (ETDEWEB)

    Tinianow, Jeff N.; Gill, Herman S.; Ogasawara, Annie; Flores, Judith E.; Vanderbilt, Alexander N.; Luis, Elizabeth; Vandlen, Richard; Darwish, Martine; Junutula, Jagath R.; Williams, Simon-P. [Genentech Research and Early Development, Genentech Inc., South San Francisco, CA 94080 (United States); Marik, Jan [Genentech Research and Early Development, Genentech Inc., South San Francisco, CA 94080 (United States)], E-mail: marik.jan@gene.com

    2010-04-15

    Three thiol reactive reagents were developed for the chemoselective conjugation of desferrioxamine (Df) to a monoclonal antibody via engineered cysteine residues (thio-trastuzumab). The in vitro stability and in vivo imaging properties of site-specifically radiolabeled {sup 89}Zr-Df-thio-trastuzumab conjugates were investigated. Methods: The amino group of desferrioxamine B was acylated by bromoacetyl bromide, N-hydroxysuccinimidyl iodoacetate, or N-hydroxysuccinimidyl 4-[N-maleimidomethyl]cyclohexane-1-carboxylate to obtain thiol reactive reagents bromoacetyl-desferrioxamine (Df-Bac), iodoacetyl-desferrioxamine (Df-Iac) and maleimidocyclohexyl-desferrioxamine (Df-Chx-Mal), respectively. Df-Bac and Df-Iac alkylated the free thiol groups of thio-trastuzumab by nucleophilic substitution forming Df-Ac-thio-trastuzumab, while the maleimide reagent Df-Chx-Mal reacted via Michael addition to provide Df-Chx-Mal-thio-trastuzumab. The conjugates were radiolabeled with {sup 89}Zr and evaluated for serum stability, and their positron emission tomography (PET) imaging properties were investigated in a BT474M1 (HER2-positive) breast tumor mouse model. Results: The chemoselective reagents were obtained in 14% (Df-Bac), 53% (Df-Iac) and 45% (Df-Chx-Mal) yields. Site-specific conjugation of Df-Chx-Mal to thio-trastuzumab was complete within 1 h at pH 7.5, while Df-Iac and Df-Bac respectively required 2 and 5 h at pH 9. Each Df modified thio-trastuzumab was chelated with {sup 89}Zr in yields exceeding 75%. {sup 89}Zr-Df-Ac-thio-trastuzumab and {sup 89}Zr-Df-Chx-Mal-thio-trastuzumab were stable in mouse serum and exhibited comparable PET imaging capabilities in a BT474M1 (HER2-positive) breast cancer model reaching 20-25 %ID/g of tumor uptake and a tumor to blood ratio of 6.1-7.1. Conclusions: The new reagents demonstrated good reactivity with engineered thiol groups of trastuzumab and very good chelation properties with {sup 89}Zr. The site-specifically {sup 89}Zr-labeled thio

  9. Site-specific labeling of proteins with NMR-active unnatural amino acids

    International Nuclear Information System (INIS)

    Jones, David H.; Cellitti, Susan E.; Hao Xueshi; Zhang Qiong; Jahnz, Michael; Summerer, Daniel; Schultz, Peter G.; Uno, Tetsuo; Geierstanger, Bernhard H.

    2010-01-01

    A large number of amino acids other than the canonical amino acids can now be easily incorporated in vivo into proteins at genetically encoded positions. The technology requires an orthogonal tRNA/aminoacyl-tRNA synthetase pair specific for the unnatural amino acid that is added to the media while a TAG amber or frame shift codon specifies the incorporation site in the protein to be studied. These unnatural amino acids can be isotopically labeled and provide unique opportunities for site-specific labeling of proteins for NMR studies. In this perspective, we discuss these opportunities including new photocaged unnatural amino acids, outline usage of metal chelating and spin-labeled unnatural amino acids and expand the approach to in-cell NMR experiments.

  10. In vivo tumor angiogenesis imaging with site-specific labeled 99mTc-HYNIC-VEGF

    International Nuclear Information System (INIS)

    Blankenberg, Francis G.; Backer, Marina V.; Patel, Vimalkumar; Backer, Joseph M.; Levashova, Zoia

    2006-01-01

    We recently developed a cysteine-containing peptide tag (C-tag) that allows for site-specific modification of C-tag-containing fusion proteins with a bifunctional chelator, HYNIC (hydrazine nicotinamide)-maleimide. We then constructed and expressed C-tagged vascular endothelial growth factor (VEGF) and labeled it with HYNIC. We wished to test 99m Tc-HYNIC-C-tagged VEGF ( 99m Tc-HYNIC-VEGF) for the imaging of tumor vasculature before and after antiangiogenic (low continuous dosing, metronomic) and tumoricidal (high-dose) cyclophosphamide treatment. HYNIC-maleimide was reacted with the two thiol groups of C-tagged VEGF without any effect on biologic activity in vitro. 99m Tc-HYNIC-VEGF was prepared using tin/tricine as an exchange reagent, and injected via the tail vein (200-300 μCi, 1-2 μg protein) followed by microSPECT imaging 1 h later. Sequencing analysis of HYNIC-containing peptides obtained after digestion confirmed the site-specific labeling of the two accessible thiol groups of C-tagged VEGF. Tumor vascularity was easily visualized with 99m Tc/VEGF in Balb/c mice with 4T1 murine mammary carcinoma 10 days after implantation into the left axillary fat pad in controls (12.3±5.0 tumor/bkg, n=27) along with its decrease following treatment with high (150 mg/kg q.o.d. x 4; 1.14±0.48 tumor/bkg, n=9) or low (25 mg/kg q.d. x 7; 1.03±0.18 tumor/bkg, n=9) dose cyclophosphamide. Binding specificity was confirmed by observing a 75% decrease in tumor uptake of 99m Tc/biotin-inactivated VEGF, as compared with 99m Tc-HYNIC-VEGF. 99m Tc can be loaded onto C-tagged VEGF in a site-specific fashion without reducing its bioactivity. 99m Tc-HYNIC-VEGF can be rapidly prepared for the imaging of tumor vasculature and its response to different types of chemotherapy. (orig.)

  11. Completion of proteomic data sets by Kd measurement using cell-free synthesis of site-specifically labeled proteins.

    Directory of Open Access Journals (Sweden)

    Paul Majkut

    Full Text Available The characterization of phosphotyrosine mediated protein-protein interactions is vital for the interpretation of downstream pathways of transmembrane signaling processes. Currently however, there is a gap between the initial identification and characterization of cellular binding events by proteomic methods and the in vitro generation of quantitative binding information in the form of equilibrium rate constants (Kd values. In this work we present a systematic, accelerated and simplified approach to fill this gap: using cell-free protein synthesis with site-specific labeling for pull-down and microscale thermophoresis (MST we were able to validate interactions and to establish a binding hierarchy based on Kd values as a completion of existing proteomic data sets. As a model system we analyzed SH2-mediated interactions of the human T-cell phosphoprotein ADAP. Putative SH2 domain-containing binding partners were synthesized from a cDNA library using Expression-PCR with site-specific biotinylation in order to analyze their interaction with fluorescently labeled and in vitro phosphorylated ADAP by pull-down. On the basis of the pull-down results, selected SH2's were subjected to MST to determine Kd values. In particular, we could identify an unexpectedly strong binding of ADAP to the previously found binding partner Rasa1 of about 100 nM, while no evidence of interaction was found for the also predicted SH2D1A. Moreover, Kd values between ADAP and its known binding partners SLP-76 and Fyn were determined. Next to expanding data on ADAP suggesting promising candidates for further analysis in vivo, this work marks the first Kd values for phosphotyrosine/SH2 interactions on a phosphoprotein level.

  12. METHOD FOR THE MEASUREMENT OF SITE-SPECIFIC TAUTOMERIC AND ZWITTERIONIC MICROSPECIES EQUILIBRIUM CONSTANTS

    Science.gov (United States)

    We describe a method for the individual measurement of simultaneously occurring, unimolecular, site-specific “microequilibrium” constants as in, for example, prototropic tautomerism and zwitterionic equilibria. Our method represents an elaboration of that of Nygren et al. (Anal. ...

  13. Site-Specific Antibody Labeling by Covalent Photoconjugation of Z Domains Functionalized for Alkyne-Azide Cycloaddition Reactions.

    Science.gov (United States)

    Perols, Anna; Arcos Famme, Melina; Eriksson Karlström, Amelie

    2015-11-01

    Antibodies are extensively used in research, diagnostics, and therapy, and for many applications the antibodies need to be labeled. Labeling is typically performed by using amine-reactive probes that target surface-exposed lysine residues, resulting in heterogeneously labeled antibodies. An alternative labeling strategy is based on the immunoglobulin G (IgG)-binding protein domain Z, which binds to the Fc region of IgG. Introducing the photoactivable amino acid benzoylphenylalanine (BPA) into the Z domain makes it possible for a covalent bond to be be formed between the Z domain and the antibody on UV irradiation, to produce a site-specifically labeled product. Z32 BPA was synthesized by solid-phase peptide synthesis and further functionalized to give alkyne-Z32 BPA and azide-Z32 BPA for Cu(I) -catalyzed cycloaddition, as well as DBCO-Z32 BPA for Cu-free strain-promoted cycloaddition. The Z32 BPA variants were conjugated to the human IgG1 antibody trastuzumab and site-specifically labeled with biotin or fluorescein. The fluorescently labeled trastuzumab showed specific staining of the membranes of HER2-expressing cells in immunofluorescence microscopy. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Differential Isotope Labeling of Glycopeptides for Accurate Determination of Differences in Site-Specific Glycosylation.

    Science.gov (United States)

    Pabst, Martin; Benešová, Iva; Fagerer, Stephan R; Jacobsen, Mathias; Eyer, Klaus; Schmidt, Gregor; Steinhoff, Robert; Krismer, Jasmin; Wahl, Fabian; Preisler, Jan; Zenobi, Renato

    2016-01-04

    We introduce a stable isotope labeling approach for glycopeptides that allows a specific glycosylation site in a protein to be quantitatively evaluated using mass spectrometry. Succinic anhydride is used to specifically label primary amino groups of the peptide portion of the glycopeptides. The heavy form (D4(13)C4) provides an 8 Da mass increment over the light natural form (H4(12)C4), allowing simultaneous analysis and direct comparison of two glycopeptide profiles in a single MS scan. We have optimized a protocol for an in-solution trypsin digestion, a one-pot labeling procedure, and a post-labeling solid-phase extraction to obtain purified and labeled glycopeptides. We provide the first demonstration of this approach by comparing IgG1 Fc glycopeptides from polyclonal IgG samples with respect to their galactosylation and sialylation patterns using MALDI MS and LC-ESI-MS.

  15. Site-Specific Protein Labeling Utilizing Lipoic Acid Ligase (LplA) and Bioorthogonal Inverse Electron Demand Diels-Alder Reaction.

    Science.gov (United States)

    Baalmann, Mathis; Best, Marcel; Wombacher, Richard

    2018-01-01

    Here, we describe a two-step protocol for selective protein labeling based on enzyme-mediated peptide labeling utilizing lipoic acid ligase (LplA) and bioorthogonal chemistry. The method can be applied to purified proteins, protein in cell lysates, as well as living cells. In a first step a W37V mutant of the lipoic acid ligase (LplA W37V ) from Escherichia coli is utilized to ligate a synthetic chemical handle site-specifically to a lysine residue in a 13 amino acid peptide motif-a short sequence that can be genetically expressed as a fusion with any protein of interest. In a second step, a molecular probe can be attached to the chemical handle in a bioorthogonal Diels-Alder reaction with inverse electron demand (DA inv ). This method is a complementary approach to protein labeling using genetic code expansion and circumvents larger protein tags while maintaining label specificity, providing experimental flexibility and straightforwardness.

  16. A method to determine site-specific, anisotropic fracture toughness in biological materials

    International Nuclear Information System (INIS)

    Bechtle, Sabine; Özcoban, Hüseyin; Yilmaz, Ezgi D.; Fett, Theo; Rizzi, Gabriele; Lilleodden, Erica T.; Huber, Norbert; Schreyer, Andreas; Swain, Michael V.; Schneider, Gerold A.

    2012-01-01

    Many biological materials are hierarchically structured, with highly anisotropic structures and properties on several length scales. To characterize the mechanical properties of such materials, detailed testing methods are required that allow precise and site-specific measurements on several length scales. We propose a fracture toughness measurement technique based on notched focused ion beam prepared cantilevers of lower and medium micron size scales. Using this approach, site-specific fracture toughness values in dental enamel were determined. The usefulness and challenges of the method are discussed.

  17. Structure-guided approach to site-specific fluorophore labeling of the lac repressor LacI.

    Directory of Open Access Journals (Sweden)

    Kalle Kipper

    Full Text Available The lactose operon repressor protein LacI has long served as a paradigm of the bacterial transcription factors. However, the mechanisms whereby LacI rapidly locates its cognate binding site on the bacterial chromosome are still elusive. Single-molecule fluorescence imaging approaches are well suited for the study of these mechanisms but rely on a functionally compatible fluorescence labeling of LacI. Particularly attractive for protein fluorescence labeling are synthetic fluorophores due to their small size and favorable photophysical characteristics. Synthetic fluorophores are often conjugated to natively occurring cysteine residues using maleimide chemistry. For a site-specific and functionally compatible labeling with maleimide fluorophores, the target protein often needs to be redesigned to remove unwanted native cysteines and to introduce cysteines at locations better suited for fluorophore attachment. Biochemical screens can then be employed to probe for the functional activity of the redesigned protein both before and after dye labeling. Here, we report a mutagenesis-based redesign of LacI to enable a functionally compatible labeling with maleimide fluorophores. To provide an easily accessible labeling site in LacI, we introduced a single cysteine residue at position 28 in the DNA-binding headpiece of LacI and replaced two native cysteines with alanines where derivatization with bulky substituents is known to compromise the protein's activity. We find that the redesigned LacI retains a robust activity in vitro and in vivo, provided that the third native cysteine at position 281 is retained in LacI. In a total internal reflection microscopy assay, we observed individual Cy3-labeled LacI molecules bound to immobilized DNA harboring the cognate O1 operator sequence, indicating that the dye-labeled LacI is functionally active. We have thus been able to generate a functional fluorescently labeled LacI that can be used to unravel mechanistic

  18. Enhanced tumor retention of a radiohalogen label for site-specific modification of antibodies.

    Science.gov (United States)

    Boswell, C Andrew; Marik, Jan; Elowson, Michael J; Reyes, Noe A; Ulufatu, Sheila; Bumbaca, Daniela; Yip, Victor; Mundo, Eduardo E; Majidy, Nicholas; Van Hoy, Marjie; Goriparthi, Saritha N; Trias, Anthony; Gill, Herman S; Williams, Simon P; Junutula, Jagath R; Fielder, Paul J; Khawli, Leslie A

    2013-12-12

    A known limitation of iodine radionuclides for labeling and biological tracking of receptor targeted proteins is the tendency of iodotyrosine to rapidly diffuse from cells following endocytosis and lysosomal degradation. In contrast, radiometal-chelate complexes such as indium-111-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (In-111-DOTA) accumulate within target cells due to the residualizing properties of the polar, charged metal-chelate-amino acid adduct. Iodine radionuclides boast a diversity of nuclear properties and chemical means for incorporation, prompting efforts to covalently link radioiodine with residualizing molecules. Herein, we describe the Ugi-assisted synthesis of [I-125]HIP-DOTA, a 4-hydroxy-3-iodophenyl (HIP) derivative of DOTA, and demonstration of its residualizing properties in a murine xenograft model. Overall, this study displays the power of multicomponent synthesis to yield a versatile radioactive probe for antibodies across multiple therapeutic areas with potential applications in both preclinical biodistribution studies and clinical radioimmunotherapies.

  19. Nanobodies: site-specific labeling for super-resolution imaging, rapid epitope-mapping and native protein complex isolation

    Science.gov (United States)

    Pleiner, Tino; Bates, Mark; Trakhanov, Sergei; Lee, Chung-Tien; Schliep, Jan Erik; Chug, Hema; Böhning, Marc; Stark, Holger; Urlaub, Henning; Görlich, Dirk

    2015-01-01

    Nanobodies are single-domain antibodies of camelid origin. We generated nanobodies against the vertebrate nuclear pore complex (NPC) and used them in STORM imaging to locate individual NPC proteins with nanobody sequence and labeled the resulting proteins with fluorophore-maleimides. As nanobodies are normally stabilized by disulfide-bonded cysteines, this appears counterintuitive. Yet, our analysis showed that this caused no folding problems. Compared to traditional NHS ester-labeling of lysines, the cysteine-maleimide strategy resulted in far less background in fluorescence imaging, it better preserved epitope recognition and it is site-specific. We also devised a rapid epitope-mapping strategy, which relies on crosslinking mass spectrometry and the introduced ectopic cysteines. Finally, we used different anti-nucleoporin nanobodies to purify the major NPC building blocks – each in a single step, with native elution and, as demonstrated, in excellent quality for structural analysis by electron microscopy. The presented strategies are applicable to any nanobody and nanobody-target. DOI: http://dx.doi.org/10.7554/eLife.11349.001 PMID:26633879

  20. Bacterial production of site specific {sup 13}C labeled phenylalanine and methodology for high level incorporation into bacterially expressed recombinant proteins

    Energy Technology Data Exchange (ETDEWEB)

    Ramaraju, Bhargavi; McFeeters, Hana; Vogler, Bernhard; McFeeters, Robert L., E-mail: robert.mcfeeters@uah.edu [University of Alabama in Huntsville, Department of Chemistry (United States)

    2017-01-15

    Nuclear magnetic resonance spectroscopy studies of ever larger systems have benefited from many different forms of isotope labeling, in particular, site specific isotopic labeling. Site specific {sup 13}C labeling of methyl groups has become an established means of probing systems not amenable to traditional methodology. However useful, methyl reporter sites can be limited in number and/or location. Therefore, new complementary site specific isotope labeling strategies are valuable. Aromatic amino acids make excellent probes since they are often found at important interaction interfaces and play significant structural roles. Aromatic side chains have many of the same advantages as methyl containing amino acids including distinct {sup 13}C chemical shifts and multiple magnetically equivalent {sup 1}H positions. Herein we report economical bacterial production and one-step purification of phenylalanine with {sup 13}C incorporation at the Cα, Cγ and Cε positions, resulting in two isolated {sup 1}H-{sup 13}C spin systems. We also present methodology to maximize incorporation of phenylalanine into recombinantly overexpressed proteins in bacteria and demonstrate compatibility with ILV-methyl labeling. Inexpensive, site specific isotope labeled phenylalanine adds another dimension to biomolecular NMR, opening new avenues of study.

  1. Applications of site-specific labeling to study HAMLET, a tumoricidal complex of α-lactalbumin and oleic acid.

    Science.gov (United States)

    Mercer, Natalia; Ramakrishnan, Boopathy; Boeggeman, Elizabeth; Qasba, Pradman K

    2011-01-01

    Alpha-lactalbumin (α-LA) is a calcium-bound mammary gland-specific protein that is found in milk. This protein is a modulator of β1,4-galactosyltransferase enzyme, changing its acceptor specificity from N-acetyl-glucosamine to glucose, to produce lactose, milk's main carbohydrate. When calcium is removed from α-LA, it adopts a molten globule form, and this form, interestingly, when complexed with oleic acid (OA) acquires tumoricidal activity. Such a complex made from human α-LA (hLA) is known as HAMLET (Human A-lactalbumin Made Lethal to Tumor cells), and its tumoricidal activity has been well established. In the present work, we have used site-specific labeling, a technique previously developed in our laboratory, to label HAMLET with biotin, or a fluoroprobe for confocal microscopy studies. In addition to full length hLA, the α-domain of hLA (αD-hLA) alone is also included in the present study. We have engineered these proteins with a 17-amino acid C-terminal extension (hLA-ext and αD-hLA-ext). A single Thr residue in this extension is glycosylated with 2-acetonyl-galactose (C2-keto-galactose) using polypeptide-α-N-acetylgalactosaminyltransferase II (ppGalNAc-T2) and further conjugated with aminooxy-derivatives of fluoroprobe or biotin molecules. We found that the molten globule form of hLA and αD-hLA proteins, with or without C-terminal extension, and with and without the conjugated fluoroprobe or biotin molecule, readily form a complex with OA and exhibits tumoricidal activity similar to HAMLET made with full-length hLA protein. The confocal microscopy studies with fluoroprobe-labeled samples show that these proteins are internalized into the cells and found even in the nucleus only when they are complexed with OA. The HAMLET conjugated with a single biotin molecule will be a useful tool to identify the cellular components that are involved with it in the tumoricidal activity.

  2. Chemoenzymatic site-specific labeling of influenza glycoproteins as a tool to observe virus budding in real time.

    Directory of Open Access Journals (Sweden)

    Maximilian Wei-Lin Popp

    Full Text Available The influenza virus uses the hemagglutinin (HA and neuraminidase (NA glycoproteins to interact with and infect host cells. While biochemical and microscopic methods allow examination of the early steps in flu infection, the genesis of progeny virions has been more difficult to follow, mainly because of difficulties inherent in fluorescent labeling of flu proteins in a manner compatible with live cell imaging. We here apply sortagging as a chemoenzymatic approach to label genetically modified but infectious flu and track the flu glycoproteins during the course of infection. This method cleanly distinguishes influenza glycoproteins from host glycoproteins and so can be used to assess the behavior of HA or NA biochemically and to observe the flu glycoproteins directly by live cell imaging.

  3. Site-Specific Hydrogen Isotope Composition of Propane: Mass spectrometric methods, equilibrium temperature dependence, and kinetics of exchange

    Science.gov (United States)

    Xie, H.; Ponton, C.; Kitchen, N.; Lloyd, M. K.; Lawson, M.; Formolo, M. J.; Eiler, J. M.

    2016-12-01

    Intramolecular isotope ordering can constrain temperatures of synthesis, mechanisms of formation, and/or source substrates of organic compounds. Here we explore site-specific hydrogen isotope variations of propane. Statistical thermodynamic models predict that at equilibrium methylene hydrogen (-CH2-) in propane will be 10's of per mil higher in D/H ratio than methyl hydrogen (-CH3) at geologically relevant temperatures, and that this difference is highly temperature dependent ( 0.5-1 ‰/°C). Chemical-kinetic controls on site-specific D/H in propane could constrain the mechanisms, conditions and extents of propane synthesis or destruction. We have developed a method for measuring the difference in D/H ratio between methylene and methyl hydrogen in propane by gas source mass spectrometry. The data were measured using the Thermo Fisher Double Focusing Sector high resolution mass spectrometer (DFS), and involve comparison of the D/H ratios of molecular ion (C3H8+) and the ethyl fragmental ion (C2H5+). We demonstrate the accuracy and precision of this method through analysis of D-labeled and independently analyzed propanes. In the exchange experiments, propane was heated (100-200 oC) either alone or in the presence of D-enriched water (δD=1,1419 ‰ SMOW), with or without one of several potentially catalytic substrates for hours to weeks. Propane was found to exchange hydrogen with water vigorously at 200 °C in the presence of metal catalysts. In the presence of Ni catalyst, methylene hydrogen exchanges 2.5 times faster than methyl hydrogen. Hydrogen exchange in the presence of Pd catalyst is more effective and can equilibrate hydrogen isotope distribution on propane on the order of 7 days. Isotopic exchange in the presence of natural materials have also been tested, but is only measurable in the methylene group at 200 °C. High catalytic activity of Pd permits attainment of a bracketed, time-invariant equilibrium state that we use to calibrate the site-specific

  4. Dual Mode Fluorophore-Doped Nickel Nitrilotriacetic Acid-Modified Silica Nanoparticles Combine Histidine-Tagged Protein Purification with Site-Specific Fluorophore Labeling

    OpenAIRE

    Kim, Sung Hoon; Jeyakumar, M.; Katzenellenbogen, John A.

    2007-01-01

    We present the first example of a fluorophore-doped nickel chelate surface- modified silica nanoparticle that functions in a dual mode, combining histidine-tagged protein purification with site-specific fluorophore labeling. Tetramethylrhodamine (TMR)-doped silica nanoparticles, estimated to contain 700–900 TMRs per ca. 23-nm particle, were surface modified with nitrilotriacetic acid (NTA), producing TMR-SiO2-NTA-Ni+2. Silica-embedded TMR retains very high quantum yield, is resistant to quenc...

  5. Site-Specific Three-Color Labeling of α-Synuclein via Conjugation to Uniquely Reactive Cysteines during Assembly by Native Chemical Ligation.

    Science.gov (United States)

    Lee, Taehyung C; Moran, Crystal R; Cistrone, Philip A; Dawson, Philip E; Deniz, Ashok A

    2018-04-12

    Single-molecule fluorescence is widely used to study conformational complexity in proteins, and has proven especially valuable with intrinsically disordered proteins (IDPs). Protein studies using dual-color single-molecule Förster resonance energy transfer (smFRET) are now quite common, but many could benefit from simultaneous measurement of multiple distances through multi-color labeling. Such studies, however, have suffered from limitations in site-specific incorporation of more than two dyes per polypeptide. Here we present a fully site-specific three-color labeling scheme for α-synuclein, an IDP with important putative functions and links to Parkinson disease. The convergent synthesis combines native chemical ligation with regiospecific cysteine protection of expressed protein fragments to permit highly controlled labeling via standard cysteine-maleimide chemistry, enabling more global smFRET studies. Furthermore, this modular approach is generally compatible with recombinant proteins and expandable to accommodate even more complex experiments, such as by labeling with additional colors. Copyright © 2018 Elsevier Ltd. All rights reserved.

  6. Dual-mode fluorophore-doped nickel nitrilotriacetic acid-modified silica nanoparticles combine histidine-tagged protein purification with site-specific fluorophore labeling.

    Science.gov (United States)

    Kim, Sung Hoon; Jeyakumar, M; Katzenellenbogen, John A

    2007-10-31

    We present the first example of a fluorophore-doped nickel chelate surface-modified silica nanoparticle that functions in a dual mode, combining histidine-tagged protein purification with site-specific fluorophore labeling. Tetramethylrhodamine (TMR)-doped silica nanoparticles, estimated to contain 700-900 TMRs per ca. 23 nm particle, were surface modified with nitrilotriacetic acid (NTA), producing TMR-SiO2-NTA-Ni2+. Silica-embedded TMR retains very high quantum yield, is resistant to quenching by buffer components, and is modestly quenched and only to a certain depth (ca. 2 nm) by surface-attached Ni2+. When exposed to a bacterial lysate containing estrogen receptor alpha ligand binding domain (ERalpha) as a minor component, these beads showed very high specificity binding, enabling protein purification in one step. The capacity and specificity of these beads for binding a his-tagged protein were characterized by electrophoresis, radiometric counting, and MALDI-TOF MS. ERalpha, bound to TMR-SiO2-NTA-Ni++ beads in a site-specific manner, exhibited good activity for ligand binding and for ligand-induced binding to coactivators in solution FRET experiments and protein microarray fluorometric and FRET assays. This dual-mode type TMR-SiO2-NTA-Ni2+ system represents a powerful combination of one-step histidine-tagged protein purification and site-specific labeling with multiple fluorophore species.

  7. Assessment of Wind Turbine for Site-Specific Conditions using Probabilistic Methods

    DEFF Research Database (Denmark)

    Heras, Enrique Gómez de las; Gutiérrez, Roberto; Azagra, Elena

    2013-01-01

    turbines, helping to the decision making during the site assessment phase of wind farm designs. First, the design equation for the failure mode of interest is defined, where the loads associated to the site-specific wind conditions are compared with the design limits of the structural component. A limit...... be very dependent on the site. The uncertainties on the wind properties depend on issues like the available wind data, the quality of the measurement sensors, the type of terrain or the accuracy of the engineering models for horizontal and vertical spatial extrapolation. An example is included showing two...

  8. Site-specific Orientation of an α-helical Peptide Ovispirin-1 from Isotope Labeled SFG Spectroscopy

    Science.gov (United States)

    Ding, Bei; Laaser, Jennifer E.; Liu, Yuwei; Wang, Pengrui; Zanni, Martin T.; Chen, Zhan

    2013-01-01

    Sum-frequency generation (SFG) vibrational spectroscopy is often used to probe the backbone structures and orientations of polypeptides at surfaces. Using the ovispirin-1 polypeptide at the solid/liquid interface of polystyrene, we demonstrate for the first time that SFG can probe the polarization response of a single isotope labeled residue. To interpret the spectral intensities, we simulated the spectra using an excitonic Hamiltonian approach. We show that the polarization dependence of either the label or the unlabeled amide I band alone does not provide sufficient structural constraints to obtain both the tilt and the twist of the ovispirin helix at a solid/liquid interface, but that both can be determined from the polarization dependence of the complete spectrum. For ovispirin, the detailed analysis of the polarized SFG experimental data shows that the helix axis is tilted at roughly 138 degrees from the surface normal, and the transition dipole of the isotope labeled C=O group is tilted at 23 degrees from the surface normal, with the hydrophobic region facing the polystyrene surface. We further demonstrated that the Hamiltonian approach is able to address the coupling effect and the structural disorder. For comparison, we also collected the FTIR spectrum of ovispirin under similar conditions, which reveals the enhanced sensitivity of SFG for structural studies of single monolayer peptide surfaces. Our study provides insight into how structural and environmental effects appear in SFG spectra of the amide I band and establishes that SFG of isotope labeled peptides will be a powerful technique for elucidating secondary structures with residue-by-residue resolution. PMID:24228619

  9. Site-specific orientation of an α-helical peptide ovispirin-1 from isotope-labeled SFG spectroscopy.

    Science.gov (United States)

    Ding, Bei; Laaser, Jennifer E; Liu, Yuwei; Wang, Pengrui; Zanni, Martin T; Chen, Zhan

    2013-11-27

    Sum-frequency generation (SFG) vibrational spectroscopy is often used to probe the backbone structures and orientations of polypeptides at surfaces. Using the ovispirin-1 polypeptide at the solid/liquid interface of polystyrene, we demonstrate for the first time that SFG can probe the polarization response of a single-isotope-labeled residue. To interpret the spectral intensities, we simulated the spectra using an excitonic Hamiltonian approach. We show that the polarization dependence of either the label or the unlabeled amide I band alone does not provide sufficient structural constraints to obtain both the tilt and the twist of the ovispirin helix at a solid/liquid interface, but that both can be determined from the polarization dependence of the complete spectrum. For ovispirin, the detailed analysis of the polarized SFG experimental data shows that the helix axis is tilted at roughly 138° from the surface normal, and the transition dipole of the isotope-labeled C═O group is tilted at 23° from the surface normal, with the hydrophobic region facing the polystyrene surface. We further demonstrate that the Hamiltonian approach is able to address the coupling effect and the structural disorder. For comparison, we also collected the FTIR spectrum of ovispirin under similar conditions, which reveals the enhanced sensitivity of SFG for structural studies of single monolayer peptide surfaces. Our study provides insight into how structural and environmental effects appear in SFG spectra of the amide I band and establishes that SFG of isotope-labeled peptides will be a powerful technique for elucidating secondary structures with residue-by-residue resolution.

  10. Site-specific quantification of lysine acetylation in the N-terminal tail of histone H4 using a double-labelling, targeted UHPLC MS/MS approach

    NARCIS (Netherlands)

    D'Urzo, Annalisa; Boichenko, Alexander P.; van den Bosch, Thea; Hermans, Jos; Dekker, Frank; Andrisano, Vincenza; Bischoff, Rainer

    We developed a targeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the site-specific quantification of lysine acetylation in the N-terminal region of histone H4 by combining chemical derivatization at the protein and peptide levels with digestion using chymotrypsin and

  11. Site-SpecificCu Labeling of the Serine Protease, Active Site Inhibited Factor Seven Azide (FVIIai-N), Using Copper Free Click Chemistry

    DEFF Research Database (Denmark)

    Jeppesen, Troels E; Kristensen, Lotte K; Nielsen, Carsten H

    2018-01-01

    A method for site-specific radiolabeling of the serine protease active site inhibited factor seven (FVIIai) with64Cu has been applied using a biorthogonal click reaction. FVIIai binds to tissue factor (TF), a trans-membrane protein involved in hemostasis, angiogenesis, proliferation, cell migrati...

  12. Introducing site-specific cysteines into nanobodies for mercury labelling allows de novo phasing of their crystal structures

    DEFF Research Database (Denmark)

    Hansen, Simon Boje; Laursen, Nick Stub; Andersen, Gregers Rom

    2017-01-01

    of the presence of free cysteines in the target protein could considerably facilitate the process of obtaining unbiased experimental phases. Nanobodies (single-domain antibodies) have recently been shown to promote the crystallization and structure determination of flexible proteins and complexes. To extend...... phased using single-wavelength anomalous dispersion (SAD) and single isomorphous replacement with anomalous signal (SIRAS), taking advantage of radiation-induced changes in Cys-Hg bonding. Importantly, Hg labelling influenced neither the interaction of Nb36 with its antigen complement C5 nor its...

  13. Neuroleptic binding sites: specific labeling in mice with [18F]haloperidol, a potential tracer for positron emission tomography

    International Nuclear Information System (INIS)

    Zanzonico, P.B.; Bigler, R.E.; Schmall, B.

    1983-01-01

    Haloperidol labeled with fluorine- 18 (T 1/2 . 110 min, positron emission 97%), prepared yielding .04 Ci/millimole by the Balz-Schiemann reaction, was evaluated in a murine model as a potential radiotracer for noninvasive determination, by positron-emission tomography, of regional concentrations of brain dopamine receptors in patients. As the haloperidol dose in mice was increased from 0.01 to 1000 micrograms/kg, the relative concentration of [ 18 F]haloperidol (microCi per g specimen/microCi per g of body mass), at one hour after injection decreased from 30 to 1.0 in the striatum and from 8.0 to 1.0 in the cerebellum. The striatal radioactivity, plotted as relative concentration against log of dose, decreased sigmoidally, presumably reflecting competition between labeled and unlabeled haloperidol for a single class of accessible binding sites. Because the cerebellum is relatively deficient in dopamine receptors, the observed decrease in cerebellar radioactivity may reflect a saturable component of haloperidol transport into brain. The high brain concentrations and the unexpectedly high striatum-to-cerebellum concentration ratios (greater than 4 at haloperidol doses less than or equal to 1 microgram/kg) suggest that [ 18 F]haloperidol warrants further investigation as a potential radiotracer for dopamine receptors

  14. Broad substrate tolerance of tubulin tyrosine ligase enables one-step site-specific enzymatic protein labeling.

    Science.gov (United States)

    Schumacher, Dominik; Lemke, Oliver; Helma, Jonas; Gerszonowicz, Lena; Waller, Verena; Stoschek, Tina; Durkin, Patrick M; Budisa, Nediljko; Leonhardt, Heinrich; Keller, Bettina G; Hackenberger, Christian P R

    2017-05-01

    The broad substrate tolerance of tubulin tyrosine ligase is the basic rationale behind its wide applicability for chemoenzymatic protein functionalization. In this context, we report that the wild-type enzyme enables ligation of various unnatural amino acids that are substantially bigger than and structurally unrelated to the natural substrate, tyrosine, without the need for extensive protein engineering. This unusual substrate flexibility is due to the fact that the enzyme's catalytic pocket forms an extended cavity during ligation, as confirmed by docking experiments and all-atom molecular dynamics simulations. This feature enabled one-step C-terminal biotinylation and fluorescent coumarin labeling of various functional proteins as demonstrated with ubiquitin, an antigen binding nanobody, and the apoptosis marker Annexin V. Its broad substrate tolerance establishes tubulin tyrosine ligase as a powerful tool for in vitro enzyme-mediated protein modification with single functional amino acids in a specific structural context.

  15. Facile method for the site-specific, covalent attachment of full-length IgG onto nanoparticles.

    Science.gov (United States)

    Hui, James Zhe; Al Zaki, Ajlan; Cheng, Zhiliang; Popik, Vladimir; Zhang, Hongtao; Luning Prak, Eline T; Tsourkas, Andrew

    2014-08-27

    Antibodies, most commonly IgGs, have been widely used as targeting ligands in research and therapeutic applications due to their wide array of targets, high specificity and proven efficacy. Many of these applications require antibodies to be conjugated onto surfaces (e.g. nanoparticles and microplates); however, most conventional bioconjugation techniques exhibit low crosslinking efficiencies, reduced functionality due to non-site-specific labeling and random surface orientation, and/or require protein engineering (e.g. cysteine handles), which can be technically challenging. To overcome these limitations, we have recombinantly expressed Protein Z, which binds the Fc region of IgG, with an UV active non-natural amino acid benzoylphenyalanine (BPA) within its binding domain. Upon exposure to long wavelength UV light, the BPA is activated and forms a covalent link between the Protein Z and the bound Fc region of IgG. This technology was combined with expressed protein ligation (EPL), which allowed for the introduction of a fluorophore and click chemistry-compatible azide group onto the C-terminus of Protein Z during the recombinant protein purification step. This enabled the crosslinked-Protein Z-IgG complexes to be efficiently and site-specifically attached to aza-dibenzocyclooctyne-modified nanoparticles, via copper-free click chemistry. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Development of an efficient signal amplification strategy for label-free enzyme immunoassay using two site-specific biotinylated recombinant proteins

    International Nuclear Information System (INIS)

    Tang, Jin-Bao; Tang, Ying; Yang, Hong-Ming

    2015-01-01

    Highlights: • An efficient signal amplification strategy for label-free EIA is proposed. • Divalent biotinylated AP and monovalent biotinylated ZZ were prepared via Avitag–BirA system. • The above site-specific biotinylated fusion proteins form complex via SA–biotin interaction. • The mechanism relies on the ZZ–Avi-B/SA/AP–(Avi-B) 2 complex. • The analytical signals are enhanced (32-fold) by the proposed strategy. - Abstract: Constructing a recombinant protein between a reporter enzyme and a detector protein to produce a homogeneous immunological reagent is advantageous over random chemical conjugation. However, the approach hardly recombines multiple enzymes in a difunctional fusion protein, which results in insufficient amplification of the enzymatic signal, thereby limiting its application in further enhancement of analytical signal. In this study, two site-specific biotinylated recombinant proteins, namely, divalent biotinylated alkaline phosphatase (AP) and monovalent biotinylated ZZ domain, were produced by employing the Avitag–BirA system. Through the high streptavidin (SA)–biotin interaction, the divalent biotinylated APs were clustered in the SA–biotin complex and then incorporated with the biotinylated ZZ. This incorporation results in the formation of a functional macromolecule that involves numerous APs, thereby enhancing the enzymatic signal, and in the production of several ZZ molecules for the interaction with immunoglobulin G (IgG) antibody. The advantage of this signal amplification strategy is demonstrated through ELISA, in which the analytical signal was substantially enhanced, with a 32-fold increase in the detection sensitivity compared with the ZZ–AP fusion protein approach. The proposed immunoassay without chemical modification can be an alternative strategy to enhance the analytical signals in various applications involving immunosensors and diagnostic chips, given that the label-free IgG antibody is suitable for

  17. Development of an efficient signal amplification strategy for label-free enzyme immunoassay using two site-specific biotinylated recombinant proteins

    Energy Technology Data Exchange (ETDEWEB)

    Tang, Jin-Bao [School of Pharmacy, Weifang Medical University, Weifang 261053 (China); Tang, Ying [Affiliated Hospital of Weifang Medical University, Weifang 261041 (China); Yang, Hong-Ming, E-mail: yanghongming2006@sohu.com [School of Pharmacy, Weifang Medical University, Weifang 261053 (China)

    2015-02-15

    Highlights: • An efficient signal amplification strategy for label-free EIA is proposed. • Divalent biotinylated AP and monovalent biotinylated ZZ were prepared via Avitag–BirA system. • The above site-specific biotinylated fusion proteins form complex via SA–biotin interaction. • The mechanism relies on the ZZ–Avi-B/SA/AP–(Avi-B){sub 2} complex. • The analytical signals are enhanced (32-fold) by the proposed strategy. - Abstract: Constructing a recombinant protein between a reporter enzyme and a detector protein to produce a homogeneous immunological reagent is advantageous over random chemical conjugation. However, the approach hardly recombines multiple enzymes in a difunctional fusion protein, which results in insufficient amplification of the enzymatic signal, thereby limiting its application in further enhancement of analytical signal. In this study, two site-specific biotinylated recombinant proteins, namely, divalent biotinylated alkaline phosphatase (AP) and monovalent biotinylated ZZ domain, were produced by employing the Avitag–BirA system. Through the high streptavidin (SA)–biotin interaction, the divalent biotinylated APs were clustered in the SA–biotin complex and then incorporated with the biotinylated ZZ. This incorporation results in the formation of a functional macromolecule that involves numerous APs, thereby enhancing the enzymatic signal, and in the production of several ZZ molecules for the interaction with immunoglobulin G (IgG) antibody. The advantage of this signal amplification strategy is demonstrated through ELISA, in which the analytical signal was substantially enhanced, with a 32-fold increase in the detection sensitivity compared with the ZZ–AP fusion protein approach. The proposed immunoassay without chemical modification can be an alternative strategy to enhance the analytical signals in various applications involving immunosensors and diagnostic chips, given that the label-free IgG antibody is suitable

  18. Separation of soil respiration: a site-specific comparison of partition methods

    Science.gov (United States)

    Comeau, Louis-Pierre; Lai, Derrick Y. F.; Jinglan Cui, Jane; Farmer, Jenny

    2018-06-01

    Without accurate data on soil heterotrophic respiration (Rh), assessments of soil carbon (C) sequestration rate and C balance are challenging to produce. Accordingly, it is essential to determine the contribution of the different sources of the total soil CO2 efflux (Rs) in different ecosystems, but to date, there are still many uncertainties and unknowns regarding the soil respiration partitioning procedures currently available. This study compared the suitability and relative accuracy of five different Rs partitioning methods in a subtropical forest: (1) regression between root biomass and CO2 efflux, (2) lab incubations with minimally disturbed soil microcosm cores, (3) root exclusion bags with hand-sorted roots, (4) root exclusion bags with intact soil blocks and (5) soil δ13C-CO2 natural abundance. The relationship between Rh and soil moisture and temperature was also investigated. A qualitative evaluation table of the partition methods with five performance parameters was produced. The Rs was measured weekly from 3 February to 19 April 2017 and found to average 6.1 ± 0.3 Mg C ha-1 yr-1. During this period, the Rh measured with the in situ mesh bags with intact soil blocks and hand-sorted roots was estimated to contribute 49 ± 7 and 79 ± 3 % of Rs, respectively. The Rh percentages estimated with the root biomass regression, microcosm incubation and δ13C-CO2 natural abundance were 54 ± 41, 8-17 and 61 ± 39 %, respectively. Overall, no systematically superior or inferior Rs partition method was found. The paper discusses the strengths and weaknesses of each technique with the conclusion that combining two or more methods optimizes Rh assessment reliability.

  19. Developing site-specific interactive environmental management tools: An exciting method of communicating training, procedures, and other information

    Energy Technology Data Exchange (ETDEWEB)

    Jaeckels, J.M.

    1999-07-01

    Environmental managers are faced with numerous programs that must be communicated throughout their organizations. Among these are regulatory training programs, internal environmental policy, regulatory guidance/procedures and internal guidance/procedures. Traditional methods of delivering this type of information are typically confined to written materials and classroom training. There are many challenges faced by environmental managers with these traditional approaches including: determining if recipients of written plans or procedures are reading and comprehending the information; scheduling training sessions to reach all affected people across multiple schedules/shifts; and maintaining adequate training records. In addition, current trends toward performance-based or competency-based training requires a more consistent method of measuring and documenting performance. The use of interactive computer applications to present training or procedural information is a new and exciting tool for delivering environmental information to employees. Site-specific pictures, text, sound, and even video can be combined with multimedia software to create informative and highly interactive applications. Some of the applications that can be produced include integrated environmental training, educational pieces, and interactive environmental procedures. They can be executed from a CD-ROM, hard drive, network or a company Intranet. Collectively, the authors refer to these as interactive environmental management tools (IEMTs). This paper focuses on site-specific, interactive training as an example of an IEMT. Interactive training not only delivers a highly effective message, but can also be designed to focus on site-specific environmental issues that are unique to each company. Interactive training also lends itself well to automated record keeping functions and to reaching all affected employees.

  20. Statistical methods for site-specific analysis of cancer among the A-bomb survivors

    International Nuclear Information System (INIS)

    Pierce, D.A.; Preston, D.L.

    1992-01-01

    Statistical methods are presented for joint, or simultaneous, analysis of the risks of several types of cancer for the A-bomb survivors. Previous analyses have been made either for all cancers except leukemia together, or have been done separately by cancer type. Either of these approaches has serious limitations, and the aim of joint analysis is to overcome these, while taking advantage of the strengths of each. The primary advantage of joint analysis is that models for risks of various cancer types can have some parameters in common, and others which are type-specific. This serves to overcome difficulties due to the limited data on specific cancer types. It also provides for significant tests comparing both type-specific risks and type-specific effects of modifying factors such as sex and age. These methods are exemplified here by joint analysis of three classes of cancer considered by the BEIR-V committee: (i) respiratory, (ii) digestive, and (iii) other cancers, excluding leukemia and breast cancer. The primary aim is to illustrate the general advantages of joint analyses, but in addition some comparison is made between the results of such joint analyses and the conclusions drawn by BEIR-V committee from separate analyses. (author)

  1. Testing of toxicity based methods to develop site specific clean up objectives - phase 1: Toxicity protocol screening and applicability

    International Nuclear Information System (INIS)

    Hamilton, H.; Kerr, D.; Thorne, W.; Taylor, B.; Zadnik, M.; Goudey, S.; Birkholz, D.

    1994-03-01

    A study was conducted to develop a cost-effective and practical protocol for using bio-assay based toxicity assessment methods for remediation of decommissioned oil and gas production, and processing facilities. The objective was to generate site-specific remediation criteria for contaminated sites. Most companies have used the chemical-specific approach which, however, did not meet the ultimate land use goal of agricultural production. The toxicity assessment method described in this study dealt with potential impairment to agricultural crop production and natural ecosystems. Human health concerns were not specifically addressed. It was suggested that chemical-specific methods should be used when human health concerns exist. . Results showed that toxicity tests will more directly identify ecological stress caused by site contamination than chemical-specific remediation criteria, which can be unnecessarily protective. 11 refs., 7 tabs., 6 figs

  2. Site Specific Vendor's License

    Data.gov (United States)

    Montgomery County of Maryland — This dataset contains information of a site-specific vendor's license which is required if an individual sells or offers to sell goods or services from a stationary...

  3. Site-Specific PEGylation of Therapeutic Proteins

    Directory of Open Access Journals (Sweden)

    Jonathan K. Dozier

    2015-10-01

    Full Text Available The use of proteins as therapeutics has a long history and is becoming ever more common in modern medicine. While the number of protein-based drugs is growing every year, significant problems still remain with their use. Among these problems are rapid degradation and excretion from patients, thus requiring frequent dosing, which in turn increases the chances for an immunological response as well as increasing the cost of therapy. One of the main strategies to alleviate these problems is to link a polyethylene glycol (PEG group to the protein of interest. This process, called PEGylation, has grown dramatically in recent years resulting in several approved drugs. Installing a single PEG chain at a defined site in a protein is challenging. Recently, there is has been considerable research into various methods for the site-specific PEGylation of proteins. This review seeks to summarize that work and provide background and context for how site-specific PEGylation is performed. After introducing the topic of site-specific PEGylation, recent developments using chemical methods are described. That is followed by a more extensive discussion of bioorthogonal reactions and enzymatic labeling.

  4. Site-Specific Innovation

    DEFF Research Database (Denmark)

    Reeh, Henrik; Hemmersam, Peter

    2015-01-01

    Currently, cities across the Northern European region are actively redeveloping their former industrial harbours. Indeed, harbours areas are essential in the long-term transition from industrial to information and experience societies; harbours are becoming sites for new businesses and residences...... question is how innovation may contribute to urban life and site-specific qualities....

  5. Linerless label device and method

    KAUST Repository

    Binladen, Abdulkari

    2016-01-14

    This apparatus and method for applying a linerless label to an end user product includes a device with a printer for printing on a face surface of a linerless label, and a release coat applicator for applying a release coat to the face surface of the label; another device including an unwinder unit (103) to unwind a roll of printed linerless label; a belt (108); a glue applicator (102) for applying glue to the belt; a nip roller (106) for contacting and applying pressure to the face surface of the linerless label such that the glue on the belt transfers to the back surface of the linerless label; at least one slitting knife 105) positioned downstream the belt and a rewinder unit (104) positioned downstream the slitting knife; and a third device which die cuts and applies the linerless label to an end user object.

  6. Site-specific conjugation and labelling of prostate antibody 7E11C5.3 (CYT-351) with technetium-99m

    International Nuclear Information System (INIS)

    Stalteri, M.A.; Mather, S.J.; Belinka, B.A.; Coughlin, D.J.; Chengazi, V.U.; Britton, K.E.

    1997-01-01

    Attachment of chelating agents to the sugar residues of antibodies for subsequent radiolabelling is an attractive approach since it may have less effect on the immunoreactivity than attachment through lysine residues, which are distributed throughout the antibody and may be present near the antigen binding site. We have attached a new hydrazide-linked chelator CYT-395 (Cytogen Corp., Princeton, N.J.) to the sugar residues of the anti-prostate monoclonal antibody 7E11C5.3 and optimised the conditions for labelling the conjugate with technetium-99m in order to compare the conjugate to 7E11C5.3 antibody labelled directly with technetium using a mercaptoethanol reduction technique. Labelling yields of 70%-90% were obtained at specific activities up to 2000 MBq/mg antibody. The stability of the technetium-labelled conjugate in plasma or to a challenge with 0.1 or 1.0 mM cysteine was similar to that of direct-labelled antibody. In nine patients with prostate cancer, the plasma clearance of the labelled conjugate followed a two-compartment model, with an average β-phase half-life of 31.4±3.9 h. The average urinary clearance at 24 h was 15.3±5.0% of the injected dose. In this group of patients there was no significant difference between the blood and urine clearance of the labelled conjugate, and the clearances of the direct-labelled antibody. (orig.). With 5 figs

  7. Linerless label device and method

    KAUST Repository

    Binladen, Abdulkari

    2016-01-01

    This apparatus and method for applying a linerless label to an end user product includes a device with a printer for printing on a face surface of a linerless label, and a release coat applicator for applying a release coat to the face surface

  8. Site specific plan

    International Nuclear Information System (INIS)

    Hutchison, J.; Jernigan, G.

    1989-12-01

    The Environmental Restoration and Waste Management Five-Year Plan (FYP) covers the period for FY 1989 through FY 1995. The plan establishes a Department of Energy -- Headquarters (DOE-HQ) agenda for cleanup and compliance against which overall progress can be measured. The FYP covers three areas: Corrective Activities, Environmental Restoration, and Waste Management Operations. Corrective Activities are those activities necessary to bring active or standby facilities into compliance with local, state, and federal environmental regulations. Environmental restoration activities include the assessment and cleanup of surplus facilities and inactive waste sites. Waste management operations includes the treatment, storage, and disposal of wastes which are generated as a result of ongoing operations. This Site Specific Plan (SSP) has been prepared by the Savannah River Site (SRS) in order to show how environmental restoration and waste management activities that were identified during the preparation of the FYP will be implemented, tracked, and reported. The SSP describes DOE Savannah River (DOE-SR) and operating contractor, Westinghouse Savannah River Company (WSRC), organizations that are responsible, for undertaking the activities identified in this plan. The SSP has been prepared in accordance with guidance received from DOE-HQ. DOE-SR is accountable to DOE-HQ for the implementation of this plan. 8 refs., 46 figs., 23 tabs

  9. Site Specific Discrete PEGylation of 124I-Labeled mCC49 Fab′ Fragments Improves Tumor MicroPET/CT Imaging in Mice

    Science.gov (United States)

    Ding, Haiming; Carlton, Michelle M.; Povoski, Stephen P.; Milum, Keisha; Kumar, Krishan; Kothandaraman, Shankaran; Hinkle, George H.; Colcher, David; Brody, Rich; Davis, Paul D.; Pokora, Alex; Phelps, Mitchell; Martin, Edward W.; Tweedle, Michael F.

    2014-01-01

    The tumor-associated glycoprotein-72 (TAG-72) antigen is highly overexpressed in various human adenocarcinomas and anti-TAG-72 monoclonal antibodies, and fragments are therefore useful as pharmaceutical targeting vectors. In this study, we investigated the effects of site-specific PEGylation with MW 2–4 kDa discrete, branched PEGylation reagents on mCC49 Fab′ (MW 50 kDa) via in vitro TAG72 binding, and in vivo blood clearance kinetics, biodistribution, and mouse tumor microPET/CT imaging. mCC49Fab′ (Fab′-NEM) was conjugated at a hinge region cysteine with maleimide-dPEG12-(dPEG24COOH)3 acid (Mal-dPEG-A), maleimide-dPEG12-(dPEG12COOH)3 acid (Mal-dPEG-B), or maleimide-dPEG12-(m-dPEG24)3 (Mal-dPEG-C), and then radiolabeled with iodine-124 (124I) in vitro radioligand binding assays and in vivo studies used TAG-72 expressing LS174T human colon carcinoma cells and xenograft mouse tumors. Conjugation of mCC49Fab′ with Mal-dPEG-A (Fab′-A) reduced the binding affinity of the non PEGylated Fab′ by 30%; however, in vivo, Fab′-A significantly lengthened the blood retention vs Fab′-NEM (47.5 vs 28.1%/ID at 1 h, 25.1 vs 8.4%/ID at 5 h, p Fab′-NEM by 70%, blood retention, microPET/CT imaging tumor signal intensity, and residual 72 h tumor concentration by 49% (3.83 ± 1.50 vs 1.97 ± 0.29%ID/g, p < 0.05) and 63% (3.83 ± 1.50 vs 1.42 ± 0.35%ID/g, p < 0.05), respectively. We conclude that remarkably subtle changes in the structure of the PEGylation reagent can create significantly altered biologic behavior. Further study is warranted of conjugates of the triple branched, negatively charged Mal-dPEG-A. PMID:24175669

  10. Methods to develop site specific spectra and a review of the important parameters that influence the spectra

    International Nuclear Information System (INIS)

    Bernreuter, D.L.

    1979-05-01

    Problems with using risk analysis methodologies to estimate the seismic hazard at a site are discussed in the context of the U.S. Nuclear Regulatory Commission's Systematic Evaluation Program (SEP). Various methodologies that may reasonably define seismic hazard are outlined. The major assumptions that can lead to significant variations in the predicted hazard are identified. Guidance is provided to appropriate choices of parameters, and possible corrections that can extend the meager earthquake data base for sites located in the eastern United States are presented. A method that incorporates various interpretations of the same data is recommended

  11. Prediction of site-specific interactions in antibody-antigen complexes: the proABC method and server.

    KAUST Repository

    Olimpieri, Pier Paolo

    2013-06-26

    MOTIVATION: Antibodies or immunoglobulins are proteins of paramount importance in the immune system. They are extremely relevant as diagnostic, biotechnological and therapeutic tools. Their modular structure makes it easy to re-engineer them for specific purposes. Short of undergoing a trial and error process, these experiments, as well as others, need to rely on an understanding of the specific determinants of the antibody binding mode. RESULTS: In this article, we present a method to identify, on the basis of the antibody sequence alone, which residues of an antibody directly interact with its cognate antigen. The method, based on the random forest automatic learning techniques, reaches a recall and specificity as high as 80% and is implemented as a free and easy-to-use server, named prediction of Antibody Contacts. We believe that it can be of great help in re-design experiments as well as a guide for molecular docking experiments. The results that we obtained also allowed us to dissect which features of the antibody sequence contribute most to the involvement of specific residues in binding to the antigen. AVAILABILITY: http://www.biocomputing.it/proABC. CONTACT: anna.tramontano@uniroma1.it or paolo.marcatili@gmail.com SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

  12. Radioisotope methods for leucocyte labelling

    International Nuclear Information System (INIS)

    Kostadinova, I.; Kovacheva, S.

    1988-01-01

    A review is made of the labelling methods with the following tracers: 3 H-thymidine, 32 P-DP, 111 In (oxine, tropolon, acetylacetone, MERC), 99m Tc (reduced 99m Tc, lypophyl 99m Tc-complexes and 99m Tc-colloids). The main diagnosis areas are mentioned: abdominal abscesses and inflammations, inflammation foci of skeleton or of implanted prosthesis; acute myocardial infarction, bacterial endocarditis, rejection of kydney transplantations or vascular grafts. It is concluded that labelled leucocytes are very reliable for noninvasive diagnosis of inflammation foci with unclear localization

  13. Radioisotope methods for leucocyte labelling

    Energy Technology Data Exchange (ETDEWEB)

    Kostadinova, I; Kovacheva, S [Meditsinska Akademiya, Sofia (Bulgaria). Katedra po Rentgenologiya i Radiologiya

    1988-01-01

    A review is made of the labelling methods with the following tracers: {sup 3}H-thymidine, {sup 32}P-DP, {sup 111}In (oxine, tropolon, acetylacetone, MERC), {sup 99m}Tc (reduced {sup 99m}Tc, lypophyl {sup 99m}Tc-complexes and {sup 99m}Tc-colloids). The main diagnosis areas are mentioned: abdominal abscesses and inflammations, inflammation foci of skeleton or of implanted prosthesis; acute myocardial infarction, bacterial endocarditis, rejection of kydney transplantations or vascular grafts. It is concluded that labelled leucocytes are very reliable for noninvasive diagnosis of inflammation foci with unclear localization.

  14. Female site-specific transposase-induced recombination: a high-efficiency method for fine mapping mutations on the X chromosome in Drosophila.

    OpenAIRE

    Marcus, Jeffrey M

    2003-01-01

    P-element transposons in the Drosophila germline mobilize only in the presence of the appropriate transposase enzyme. Sometimes, instead of mobilizing completely, P elements will undergo site-specific recombination with the homologous chromosome. Site-specific recombination is the basis for male recombination mapping, since the male germline does not normally undergo recombination. Site-specific recombination also takes place in females, but this has been difficult to study because of the obs...

  15. Statistical and Economic Techniques for Site-specific Nematode Management.

    Science.gov (United States)

    Liu, Zheng; Griffin, Terry; Kirkpatrick, Terrence L

    2014-03-01

    Recent advances in precision agriculture technologies and spatial statistics allow realistic, site-specific estimation of nematode damage to field crops and provide a platform for the site-specific delivery of nematicides within individual fields. This paper reviews the spatial statistical techniques that model correlations among neighboring observations and develop a spatial economic analysis to determine the potential of site-specific nematicide application. The spatial econometric methodology applied in the context of site-specific crop yield response contributes to closing the gap between data analysis and realistic site-specific nematicide recommendations and helps to provide a practical method of site-specifically controlling nematodes.

  16. METHOD AND MODULE FOR OPTICAL SUBCARRIER LABELLING

    DEFF Research Database (Denmark)

    2004-01-01

    The present invention relates to optical labelling in WDM networks, in that it provides a method and a module to be used in subcarrier label generation and switching in network edge nodes and core switch nodes. The methods and modules are typically employed in Optical Subcarrier Multiplexing (OSCM......) transmitters. The payload and the label are encoded independently on optical carrier and subcarrier signals respectively, using electro-optical modulators. The invention applies single or double sideband carrier-suppressed modulation to generate subcarrier signals for encoding of the label. Thereby the payload...... encoded carrier signal and the label encoded subcarrier signal can be coupled directly without prior filtering....

  17. Bifunctional chelating agent for the design and development of site specific radiopharmaceuticals and biomolecule conjugation strategy

    Science.gov (United States)

    Katti, Kattesh V.; Prabhu, Kandikere R.; Gali, Hariprasad; Pillarsetty, Nagavara Kishore; Volkert, Wynn A.

    2003-10-21

    There is provided a method of labeling a biomolecule with a transition metal or radiometal in a site specific manner to produce a diagnostic or therapeutic pharmaceutical compound by synthesizing a P.sub.2 N.sub.2 -bifunctional chelating agent intermediate, complexing the intermediate with a radio metal or a transition metal, and covalently linking the resulting metal-complexed bifunctional chelating agent with a biomolecule in a site specific manner. Also provided is a method of synthesizing the --PR.sub.2 containing biomolecules by synthesizing a P.sub.2 N.sub.2 -bifunctional chelating agent intermediate, complexing the intermediate with a radiometal or a transition metal, and covalently linking the resulting radio metal-complexed bifunctional chelating agent with a biomolecule in a site specific manner. There is provided a therapeutic or diagnostic agent comprising a --PR.sub.2 containing biomolecule.

  18. Nanoparticles for Site Specific Genome Editing

    Science.gov (United States)

    McNeer, Nicole Ali

    Triplex-forming peptide nucleic acids (PNAs) can be used to coordinate the recombination of short 50-60 by "donor DNA" fragments into genomic DNA, resulting in site-specific correction of genetic mutations or the introduction of advantageous genetic modifications. Site-specific gene editing in hematopoietic stem and progenitor cells (HSPCs) could result in treatment or cure of inherited disorders of the blood such as beta-thalassemia. Gene editing in HSPCs and differentiated T cells could help combat HIV/AIDs by modifying receptors, such as CCR5, necessary for R5-tropic HIV entry. However, translation of genome modification technologies to clinical practice is limited by challenges in intracellular delivery, especially in difficult-to-transfect hematolymphoid cells. In vivo gene editing could also provide novel treatment for systemic monogenic disorders such as cystic fibrosis, an autosomal recessive disorder caused by mutations in the cystic fibrosis transmembrane receptor. Here, we have engineered biodegradable nanoparticles to deliver oligonucleotides for site-specific genome editing of disease-relevant genes in human cells, with high efficiency, low toxicity, and editing of clinically relevant cell types. We designed nanoparticles to edit the human beta-globin and CCR5 genes in hematopoietic cells. We show that poly(lactic-co-glycolic acid) (PLGA) nanoparticles can delivery PNA and donor DNA for site-specific gene modification in human hematopoietic cells in vitro and in vivo in NOD-scid IL2rgammanull mice. Nanoparticles delivered by tail vein localized to hematopoietic compartments in the spleen and bone marrow of humanized mice, resulting in modification of the beta-globin and CCR5 genes. Modification frequencies ranged from 0.005 to 20% of cells depending on the organ and cell type, without detectable toxicity. This project developed highly versatile methods for delivery of therapeutics to hematolymphoid cells and hematopoietic stem cells, and will help to

  19. Multiple tag labeling method for DNA sequencing

    Science.gov (United States)

    Mathies, R.A.; Huang, X.C.; Quesada, M.A.

    1995-07-25

    A DNA sequencing method is described which uses single lane or channel electrophoresis. Sequencing fragments are separated in the lane and detected using a laser-excited, confocal fluorescence scanner. Each set of DNA sequencing fragments is separated in the same lane and then distinguished using a binary coding scheme employing only two different fluorescent labels. Also described is a method of using radioisotope labels. 5 figs.

  20. Evaluation of three methods of platelet labelling

    International Nuclear Information System (INIS)

    Mortelmans, L.; Verbruggen, A.; Roo, M. de; Vermylen, J.

    1986-01-01

    The study of the kinetics of labelled platelets makes sense only when the platelets preserve their viability after separation and labelling. The separation and labelling procedures described in the manual of two producers of 111 In-oxinate (Amersham, Mallinckrodt) have been evaluated by in vitro aggregation tests. The method of Mallinckrodt diminished the aggregation capacities of the thrombocytes. The labelled platelets with normal in vitro aggregation response (Amersham) were tested in vivo in 11 patients who underwent peripheral bypass surgery. The platelet half-life and the platelet accumulation on bypass grafts were checked one week post-operatively. Because of the poor in vivo response of both methods (exponential half-life curve and bad graft visualization), a third method based on that described by W.A. Heaton et al. 1979 was optimized in the authors' laboratory with good in vitro and in vivo results in 12 patients. (author)

  1. Evaluation of three methods of platelet labelling.

    Science.gov (United States)

    Mortelmans, L; Verbruggen, A; De Roo, M; Vermylen, J

    1986-07-01

    The study of the kinetics of labelled platelets makes sense only when the platelets preserve their viability after separation and labelling. The separation and labelling procedures described in the manual of two producers of 111In-oxinate (Amersham, Mallinckrodt) have been evaluated by in vitro aggregation tests. The method of Mallinckrodt diminished the aggregation capacities of the thrombocytes. The labelled platelets with normal in vitro aggregation response (Amersham) were tested in vivo in 11 patients who underwent peripheral bypass surgery. The platelet half-life and the platelet accumulation on bypass grafts were checked one week post-operatively. Because of the poor in vivo response of both methods (exponential half-life curve and bad graft visualization), a third method was optimized in our laboratory with good in vitro and in vivo results in 12 patients.

  2. Reductive methods for isotopic labeling of antibiotics

    International Nuclear Information System (INIS)

    Champney, W.S.

    1989-01-01

    Methods for the reductive methylation of the amino groups of eight different antibiotics using 3 HCOH or H 14 COH are presented. The reductive labeling of an additional seven antibiotics by NaB 3 H 4 is also described. The specific activity of the methyl-labeled drugs was determined by a phosphocellulose paper binding assay. Two quantitative assays for these compounds based on the reactivity of the antibiotic amino groups with fluorescamine and of the aldehyde and ketone groups with 2,4-dinitrophenylhydrazine are also presented. Data on the cellular uptake and ribosome binding of these labeled compounds are also presented

  3. Position for Site-Specific Attachment of a DOTA Chelator to Synthetic Affibody Molecules Has a Different Influence on the Targeting Properties of 68Ga-Compared to 111In-Labeled Conjugates

    Directory of Open Access Journals (Sweden)

    Hadis Honarvar

    2014-12-01

    Full Text Available Affibody molecules, small (7 kDa scaffold proteins, are a promising class of probes for radionuclide molecular imaging. Radiolabeling of Affibody molecules with the positron-emitting nuclide 68Ga would permit the use of positron emission tomography (PET, providing better resolution, sensitivity, and quantification accuracy than single-photon emission computed tomography (SPECT. The synthetic anti-HER2 ZHER2:S1 Affibody molecule was conjugated with DOTA at the N-terminus, in the middle of helix 3, or at the C-terminus. The biodistribution of 68Ga- and 111In-labeled Affibody molecules was directly compared in NMRI nu/nu mice bearing SKOV3 xenografts. The position of the chelator strongly influenced the biodistribution of the tracers, and the influence was more pronounced for 68Ga-labeled Affibody molecules than for the 111In-labeled counterparts. The best 68Ga-labeled variant was 68Ga-[DOTA-A1]-ZHER2:S1 which provided a tumor uptake of 13 ± 1 %ID/g and a tumor to blood ratio of 39 ± 12 at 2 hours after injection. 111In-[DOTA-A1]-ZHER2:S1 and 111In-[DOTA-K58]-ZHER2:S1 were equally good at this time point, providing a tumor uptake of 15 to 16 %ID/g and a tumor to blood ratio in the range of 60 to 80. In conclusion, the selection of the best position for a chelator in Affibody molecules can be used for optimization of their imaging properties. This may be important for the development of Affibody-based and other protein-based imaging probes.

  4. Site-specific weed control technologies

    DEFF Research Database (Denmark)

    Christensen, Svend; Søgaard, Henning Tangen; Kudsk, Per

    2009-01-01

    Site-specific weed control technologies are defined as machinery or equipment embedded with technologies that detect weeds growing in a crop and, taking into account predefined factors such as economics, takes action to maximise the chances of successfully controlling them. In the article, we...... describe the basic parts of site specific weed control technologies, comprising of weed sensing systems, weed management models and precision weed control implements. A review of state-of-the-art technologies shows that several weed sensing systems and precision implements have been developed over the last...... of knowledge about the economic and environmental potential for increasing the resolution of weed control. The integration of site-specific information on weed distribution, weed species composition and density, and the effect on crop yield, is decisive for successful site-specific weed management.   Keywords...

  5. Comparative evaluation of synthetic anti-HER2 Affibody molecules site-specifically labelled with {sup 111}In using N-terminal DOTA, NOTA and NODAGA chelators in mice bearing prostate cancer xenografts

    Energy Technology Data Exchange (ETDEWEB)

    Malmberg, Jennie; Varasteh, Zohreh; Orlova, Anna [Uppsala University, Preclinical PET Platform, Department of Medicinal Chemistry, Uppsala (Sweden); Perols, Anna; Braun, Alexis; Eriksson Karlstroem, Amelie [AlbaNova University Centre, Division of Molecular Biotechnology, School of Biotechnology, KTH Royal Institute of Technology, Stockholm (Sweden); Altai, Mohamed; Tolmachev, Vladimir [Uppsala University, Unit of Biomedical Radiation Sciences, Rudbeck Laboratory, Uppsala (Sweden); Sandstroem, Mattias [Uppsala University Hospital, Section of Medical Physics, Department of Oncology, Uppsala (Sweden); Garske, Ulrike [Uppsala University Hospital, Department of Medical Sciences, Section of Nuclear Medicine, Uppsala (Sweden)

    2012-03-15

    In disseminated prostate cancer, expression of human epidermal growth factor receptor type 2 (HER2) is one of the pathways to androgen independence. Radionuclide molecular imaging of HER2 expression in disseminated prostate cancer might identify patients for HER2-targeted therapy. Affibody molecules are small (7 kDa) targeting proteins with high potential as tracers for radionuclide imaging. The goal of this study was to develop an optimal Affibody-based tracer for visualization of HER2 expression in prostate cancer. A synthetic variant of the anti-HER2 Z{sub HER2:342} Affibody molecule, Z{sub HER2:S1}, was N-terminally conjugated with the chelators DOTA, NOTA and NODAGA. The conjugated proteins were biophysically characterized by electrospray ionization mass spectroscopy (ESI-MS), circular dichroism (CD) spectroscopy and surface plasmon resonance (SPR)-based biosensor analysis. After labelling with {sup 111}In, the biodistribution was assessed in normal mice and the two most promising conjugates were further evaluated for tumour targeting in mice bearing DU-145 prostate cancer xenografts. The HER2-binding equilibrium dissociation constants were 130, 140 and 90 pM for DOTA-Z{sub HER2:S1}, NOTA-Z{sub HER2:S1} and NODAGA-Z{sub HER2:S1}, respectively. A comparative study of {sup 111}In-labelled DOTA-Z{sub HER2:S1}, NOTA-Z{sub HER2:S1} and NODAGA-Z{sub HER2:S1} in normal mice demonstrated a substantial influence of the chelators on the biodistribution properties of the conjugates. {sup 111}In-NODAGA-Z{sub HER2:S1} had the most rapid clearance from blood and healthy tissues. {sup 111}In-NOTA-Z{sub HER2:S1} showed high hepatic uptake and was excluded from further evaluation. {sup 111}In-DOTA-Z{sub HER2:S1} and {sup 111}In-NODAGA-Z{sub HER2:S1} demonstrated specific uptake in DU-145 prostate cancer xenografts in nude mice. The tumour uptake of {sup 111}In-NODAGA-Z{sub HER2:S1}, 5.6 {+-} 0.4%ID/g, was significantly lower than the uptake of {sup 111}In-DOTA-Z{sub HER2:S1

  6. Site-Specific Infrared Probes of Proteins

    Science.gov (United States)

    Ma, Jianqiang; Pazos, Ileana M.; Zhang, Wenkai; Culik, Robert M.; Gai, Feng

    2015-01-01

    Infrared spectroscopy has played an instrumental role in studying a wide variety of biological questions. However, in many cases it is impossible or difficult to rely on the intrinsic vibrational modes of biological molecules of interest, such as proteins, to reveal structural and/or environmental information in a site-specific manner. To overcome this limitation, many recent efforts have been dedicated to the development and application of various extrinsic vibrational probes that can be incorporated into biological molecules and used to site-specifically interrogate their structural and/or environmental properties. In this Review, we highlight some recent advancements of this rapidly growing research area. PMID:25580624

  7. DOE site-specific threat assessment

    International Nuclear Information System (INIS)

    West, D.J.; Al-Ayat, R.A.; Judd, B.R.

    1985-01-01

    A facility manager faced with the challenges of protecting a nuclear facility against potential threats must consider the likelihood and consequences of such threats, know the capabilities of the facility safeguards and security systems, and make informed decisions about the cost-effectivness of safeguards and security upgrades. To help meet these challenges, the San Francisco Operations Office of the Department of Energy, in conjunction with the Lawrence Livermore Laboratory, has developed a site-specific threat assessment approach and a quantitative model to improve the quality and consistency of site-specific threat assessment and resultant security upgrade decisions at sensitive Department of Energy facilities. 5 figs

  8. Prospects for site specific weed management

    DEFF Research Database (Denmark)

    Christensen, Svend; Rasmussen, Jesper; Pedersen, Søren Marcus

    2014-01-01

    Research on Site Specific Weed Management (SSWM) started in the late 80's. Since that moment, considerable research has been conducted on different aspects of SSWM, from fundamental studies on the spatial ecology of weeds to the applied development and testing of new technologies for weed detection...

  9. [Progress in stable isotope labeled quantitative proteomics methods].

    Science.gov (United States)

    Zhou, Yuan; Shan, Yichu; Zhang, Lihua; Zhang, Yukui

    2013-06-01

    Quantitative proteomics is an important research field in post-genomics era. There are two strategies for proteome quantification: label-free methods and stable isotope labeling methods which have become the most important strategy for quantitative proteomics at present. In the past few years, a number of quantitative methods have been developed, which support the fast development in biology research. In this work, we discuss the progress in the stable isotope labeling methods for quantitative proteomics including relative and absolute quantitative proteomics, and then give our opinions on the outlook of proteome quantification methods.

  10. Technetium-99m labeled radiodiagnostic agents and method of preparation

    International Nuclear Information System (INIS)

    1976-01-01

    A method of preparing improved technetium-99m labelled radiodiagnostic agents by reducing sup(99m)Tc-pertechnetate with stannous tartrate is given. Human serum albumine (HSA) and 1-hydroxyethylidene-1,1-disodiumphosphonate (HEDSPA), which are useful in scintigraphic examinations of the lung and bone, were labelled in this way

  11. Stem cell monitoring with a direct or indirect labeling method

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Min Hwan; Lee, Yong Jin [Molecular Imaging Research Center, Korea Institute of Radiological and Medical Sciences (KIRAMS), Seoul (Korea, Republic of)

    2016-12-15

    The molecular imaging techniques allow monitoring of the transplanted cells in the same individuals over time, from early localization to the survival, migration, and differentiation. Generally, there are two methods of stem cell labeling: direct and indirect labeling methods. The direct labeling method introduces a labeling agent into the cell, which is stably incorporated or attached to the cells prior to transplantation. Direct labeling of cells with radionuclides is a simple method with relatively fewer adverse events related to genetic responses. However, it can only allow short-term distribution of transplanted cells because of the decreasing imaging signal with radiodecay, according to the physical half-lives, or the signal becomes more diffuse with cell division and dispersion. The indirect labeling method is based on the expression of a reporter gene transduced into the cell before transplantation, which is then visualized upon the injection of an appropriate probe or substrate. In this review, various imaging strategies to monitor the survival and behavior change of transplanted stem cells are covered. Taking these new approaches together, the direct and indirect labeling methods may provide new insights on the roles of in vivo stem cell monitoring, from bench to bedside.

  12. Savannah River Site's Site Specific Plan

    International Nuclear Information System (INIS)

    1991-01-01

    This Site Specific Plan (SSP) has been prepared by the Savannah River Site (SRS) in order to show the Environmental Restoration and Waste Management activities that were identified during the preparation of the Department of Energy-Headquarters (DOE-HQ) Environmental Restoration and Waste Management Five-Year Plan (FYP) for FY 1992--1996. The SSP has been prepared in accordance with guidance received from DOE-HQ. DOE-SR is accountable to DOE-HQ for the implementation of this plan. The purpose of the SSP is to develop a baseline for policy, budget, and schedules for the DOE Environmental Restoration and Waste Management activities. The plan explains accomplishments since the Fiscal Year (FY) 1990 plan, demonstrates how present and future activities are prioritized, identifies currently funded activities and activities that are planned to be funded in the upcoming fiscal year, and describes future activities that SRS is considering

  13. Temporally-controlled site-specific recombination in zebrafish.

    Directory of Open Access Journals (Sweden)

    Stefan Hans

    Full Text Available Conventional use of the site-specific recombinase Cre is a powerful technology in mouse, but almost absent in other vertebrate model organisms. In zebrafish, Cre-mediated recombination efficiency was previously very low. Here we show that using transposon-mediated transgenesis, Cre is in fact highly efficient in this organism. Furthermore, temporal control of recombination can be achieved by using the ligand-inducible CreER(T2. Site-specific recombination only occurs upon administration of the drug tamoxifen (TAM or its active metabolite, 4-hydroxy-tamoxifen (4-OHT. Cre-mediated recombination is detectable already 4 or 2 hours after administration of TAM or 4-OHT, demonstrating fast recombination kinetics. In addition, low doses of TAM allow mosaic labeling of single cells. Combined, our results show that conditional Cre/lox will be a valuable tool for both, embryonic and adult zebrafish studies. Furthermore, single copy insertion transgenesis of Cre/lox constructs suggest a strategy suitable also for other organisms.

  14. SITE-94. Site specific base data for the performance assessment

    International Nuclear Information System (INIS)

    Geier, J.; Tiren, S.; Dverstorp, B.; Glynn, P.

    1996-06-01

    This report documents the site specific base data that were available, and the utilization of these data within SITE-94. A brief summary is given of SKB's preliminary site investigations for the Aespoe Hard Rock Laboratory (HRL), which were the main source of site-specific data for SITE-94, and an overview is given of the field methods and instrumentation for the preliminary investigations. A compilation is given of comments concerning the availability and quality of the data for Aespoe, and specific recommendations are given for future site investigations. It was found that the HRL pre-investigations produced a large quantity of data which were, for the most part, of sufficient quality to be valuable for a performance assessment. However, some problems were encountered regarding documentation, procedural consistency, positional information, and storage of the data from the measurements. 77 refs, 4 tabs

  15. Exploration of new tritium labelling methods

    International Nuclear Information System (INIS)

    Andres, H.; Jaiswal, D.K.; Morimoto, H.; Saljoughian, M.; Than, C.; Willimas, P.G.; Zippi, E.M.

    1997-01-01

    Full text: A great deal of elegant chemistry is available for hydride transfer reactions, and could be adapted for tritium labelling. Nevertheless, most high level tritiation reactions still involve either hydrogenation (alkene or alkyne precursor) or catalytic dehalogenation. In the last decade we have endeavored to propose and popularize alternative labelling techniques and reagents, including: i) the synthesis of new precursors for the production of methyl iodide; ii) the synthesis of methylene diiodide; iii) the production and use of T 2 O, and solvents made from it, eg. CH 3 COOT, CF 3 COOT; iv) high specific activity hydride reagents, eg. LiAIT 4 , LiEt 3 BT; (Bu n ) 3 SnT, ZrCp 2 CIT, LiT, Li(OCH 3 ) 3 BT, Ph 2 SiT 2 , BT 3 -THF, Li/Na/KBT 4 ; v) reduction with diimide; vi) use of T 2 O in special reactions such as the Shapiro reaction and Brook rearrangement; and vii) developments of a new acetylation reagent. We have also initiated and continued a number of innovative applications of tritium NMR spectroscopy. Many of these projects have grown out of User or Collaborator requirements at the NTLF. We regard this stimulus to develop and refine both tritiation and NMR techniques as healthy and challenging

  16. Versatile and Efficient Site-Specific Protein Functionalization by Tubulin Tyrosine Ligase.

    Science.gov (United States)

    Schumacher, Dominik; Helma, Jonas; Mann, Florian A; Pichler, Garwin; Natale, Francesco; Krause, Eberhard; Cardoso, M Cristina; Hackenberger, Christian P R; Leonhardt, Heinrich

    2015-11-09

    A novel chemoenzymatic approach for simple and fast site-specific protein labeling is reported. Recombinant tubulin tyrosine ligase (TTL) was repurposed to attach various unnatural tyrosine derivatives as small bioorthogonal handles to proteins containing a short tubulin-derived recognition sequence (Tub-tag). This novel strategy enables a broad range of high-yielding and fast chemoselective C-terminal protein modifications on isolated proteins or in cell lysates for applications in biochemistry, cell biology, and beyond, as demonstrated by the site-specific labeling of nanobodies, GFP, and ubiquitin. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Technetium-99m labeled radiodiagnostic agents and method of preparation

    International Nuclear Information System (INIS)

    Molinski, V.J.; Wilczewski, J.A.

    1977-01-01

    A method of preparing improved technetium-99m labeled radiodiagnostic agents by reducing technetium-99m with stannous tartrate is described. Such radiodiagnostic agents are useful in scintigraphic examinations of the bone and lung

  18. Recent advances in covalent, site-specific protein immobilization [version 1; referees: 3 approved

    Directory of Open Access Journals (Sweden)

    Morten Meldal

    2016-09-01

    Full Text Available The properties of biosensors, biomedical implants, and other materials based on immobilized proteins greatly depend on the method employed to couple the protein molecules to their solid support. Covalent, site-specific immobilization strategies are robust and can provide the level of control that is desired in this kind of application. Recent advances include the use of enzymes, such as sortase A, to couple proteins in a site-specific manner to materials such as microbeads, glass, and hydrogels. Also, self-labeling tags such as the SNAP-tag can be employed. Last but not least, chemical approaches based on bioorthogonal reactions, like the azide–alkyne cycloaddition, have proven to be powerful tools. The lack of comparative studies and quantitative analysis of these immobilization methods hampers the selection process of the optimal strategy for a given application. However, besides immobilization efficiency, the freedom in selecting the site of conjugation and the size of the conjugation tag and the researcher’s expertise regarding molecular biology and/or chemical techniques will be determining factors in this regard.

  19. Site specific information in site selection

    International Nuclear Information System (INIS)

    Aeikaes, T.; Hautojaervi, A.

    1998-01-01

    The programme for the siting of a deep repository for final disposal of spent nuclear fuel was started already in 1983 and is carried out today by Posiva Oy which continues the work started by Teollisuuden Voima Oy. The programme aims at site selection by the end of the year 2000. The programme has progressed in successive interim stages with defined goals. After an early phase for site identification, five sites were selected in 1987 for preliminary site characterisation. Three of these were selected and judged to be best suited for the more detailed characterisation in 1992. An additional new site was included into the programme based on a separate feasibility study in the beginning of 1997. Since the year 1983 several safety assessments together with technical plans of the facility have been completed. When approaching the site selection the needs for more detailed consideration of the site specific properties in the safety assessment have been increased. The Finnish regulator STUK has published a proposal for general safety requirements for the final disposal of spent nuclear fuel in Finland. This set of requirements has been projected to be used in conjunction of the decision making by the end 2000. Based on the site evaluation all sites can provide a stable environment and there is evidence that the requirements for the longevity of the canister can be fulfilled at each site. In this manner the four candidate sites do not differ too much from each other. The main difference between the sites is in the salinity of the deep groundwater. The significance of differences in the salinity for the long-term safety cannot be defined yet. The differences may contribute to the discussion of the longevity of the bentonite buffer and also to the modelling of the groundwater flow and transport. The use of the geosphere as a transport barrier is basically culminated on the questions about sparse but fast flow routes and 'how bad channeling can be'. To answer these questions

  20. A diagram retrieval method with multi-label learning

    Science.gov (United States)

    Fu, Songping; Lu, Xiaoqing; Liu, Lu; Qu, Jingwei; Tang, Zhi

    2015-01-01

    In recent years, the retrieval of plane geometry figures (PGFs) has attracted increasing attention in the fields of mathematics education and computer science. However, the high cost of matching complex PGF features leads to the low efficiency of most retrieval systems. This paper proposes an indirect classification method based on multi-label learning, which improves retrieval efficiency by reducing the scope of compare operation from the whole database to small candidate groups. Label correlations among PGFs are taken into account for the multi-label classification task. The primitive feature selection for multi-label learning and the feature description of visual geometric elements are conducted individually to match similar PGFs. The experiment results show the competitive performance of the proposed method compared with existing PGF retrieval methods in terms of both time consumption and retrieval quality.

  1. Taxon- and Site-Specific Melatonin Catabolism

    Directory of Open Access Journals (Sweden)

    Rüdiger Hardeland

    2017-11-01

    Full Text Available Melatonin is catabolized both enzymatically and nonenzymatically. Nonenzymatic processes mediated by free radicals, singlet oxygen, other reactive intermediates such as HOCl and peroxynitrite, or pseudoenzymatic mechanisms are not species- or tissue-specific, but vary considerably in their extent. Higher rates of nonenzymatic melatonin metabolism can be expected upon UV exposure, e.g., in plants and in the human skin. Additionally, melatonin is more strongly nonenzymatically degraded at sites of inflammation. Typical products are several hydroxylated derivatives of melatonin and N1-acetyl-N2-formyl-5-methoxykynuramine (AFMK. Most of these products are also formed by enzymatic catalysis. Considerable taxon- and site-specific differences are observed in the main enzymatic routes of catabolism. Formation of 6-hydroxymelatonin by cytochrome P450 subforms are prevailing in vertebrates, predominantly in the liver, but also in the brain. In pineal gland and non-mammalian retina, deacetylation to 5-methoxytryptamine (5-MT plays a certain role. This pathway is quantitatively prevalent in dinoflagellates, in which 5-MT induces cyst formation and is further converted to 5-methoxyindole-3-acetic acid, an end product released to the water. In plants, the major route is catalyzed by melatonin 2-hydroxylase, whose product is tautomerized to 3-acetamidoethyl-3-hydroxy-5-methoxyindolin-2-one (AMIO, which exceeds the levels of melatonin. Formation and properties of various secondary products are discussed.

  2. A convenient method to synthesize specifically labelled cholesterol with tritium

    International Nuclear Information System (INIS)

    Malik, S.; Kenny, M.; Ahmad, S.; Washington Univ., Seattle, WA

    1992-01-01

    A simple method is described to label cholesterol with tritium. Cholesterol was first oxidized to 5-cholesten-3-one which was then purified by HPLC. Its structure was established by electron impact (EI) mass spectrometry and 1 H-NMR spectroscopy. The ketone was reduced with NaB 3 H 4 to give specifically labelled cholesterol (C-3 3 H) at low specific activity. (author)

  3. Development of site specific response spectra

    International Nuclear Information System (INIS)

    Bernreuter, D.L.; Chen, J.C.; Savy, J.B.

    1987-03-01

    For a number of years the US Nuclear Regulatory Commission (NRC) has employed site specific spectra (SSSP) in their evaluation of the adequacy of the Safe Shutdown Earthquake (SSE). These spectra were developed only from the spectra of the horizontal components of the ground motion and from a very limited data set. As the data set has considerably increased for Eastern North America (ENA) and as more relevant data has become available from earthquakes occurring in other parts of the world (e.g., Italy), together with the fact that recent data indicated the importance of the vertical component, it became clear that an update of the SSSP's for ENA was desirable. The methodology used in this study is similar to the previous ones in that it used actual earthquake ground motion data with magnitudes within a certain range and recorded at distances and at sites similar to those that would be chosen for the definition of an SSE. An extensive analysis of the origin and size of the uncertainty is an important part of this study. The results of this analysis of the uncertainties is used to develop criteria for selecting the earthquake records to be used in the derivation of the SSSP's. We concluded that the SSSPs were not very sensitive to the distribution of the source to site distance of the earthquake records used in the analysis. That is, the variability (uncertainty) introduced by the range of distances was relatively small compared to the variability introduced by other factors. We also concluded that the SSSP are somewhat sensitive to the distribution of the magnitudes of these earthquakes, particularly at rock sites and, by inference, at shallow soil sites. We found that one important criterion in selecting records to generate SSSP is the depth of soil at the site

  4. Site-Specific, Climate-Friendly Farming

    Science.gov (United States)

    Brown, D. J.; Brooks, E. S.; Eitel, J.; Huggins, D. R.; Painter, K.; Rupp, R.; Smith, J. L.; Stockle, C.; Vierling, L. A.

    2011-12-01

    Of the four most important atmospheric greenhouse gasses (GHG) enriched through human activities, only nitrous oxide (N2O) emissions are due primarily to agriculture. However, reductions in the application of synthetic N fertilizers could have significant negative consequences for a growing world population given the crucial role that these fertilizers have played in cereal yield increases since WWII. Increasing N use efficiency (NUE) through precision management of agricultural N in space and time will therefore play a central role in the reduction of agricultural N2O emissions. Precision N management requires a greater understanding of the spatio-temporal variability of factors supporting N management decisions such as crop yield, water and N availability, utilization and losses. We present an overview of a large, collaborative, multi-disciplinary project designed to improve our basic understanding of nitrogen (N), carbon (C) and water (H2O) spatio-temporal dynamics for wheat-based cropping systems on complex landscapes, and develop management tools to optimize water- and nitrogen-use efficiency for these systems and landscapes. Major components of this project include: (a) cropping systems experiments addressing nitrogen application rate and seeding density for different landscape positions; (b) GHG flux experiments and monitoring; (c) soil microbial genetics and stable isotope analyses to elucidate biochemical pathways for N2O production; (d) proximal soil sensing for construction of detailed soil maps; (e) LiDAR and optical remote sensing for crop growth monitoring; (f) hydrologic experiments, monitoring, and modeling; (g) refining the CropSyst simulation model to estimate biophysical processes and GHG emissions under a variety of management and climatic scenarios; and (h) linking farm-scale enterprise budgets to simulation modeling in order to provide growers with economically viable site-specific climate-friendly farming guidance.

  5. Inter-labeler and intra-labeler variability of condition severity classification models using active and passive learning methods.

    Science.gov (United States)

    Nissim, Nir; Shahar, Yuval; Elovici, Yuval; Hripcsak, George; Moskovitch, Robert

    2017-09-01

    Labeling instances by domain experts for classification is often time consuming and expensive. To reduce such labeling efforts, we had proposed the application of active learning (AL) methods, introduced our CAESAR-ALE framework for classifying the severity of clinical conditions, and shown its significant reduction of labeling efforts. The use of any of three AL methods (one well known [SVM-Margin], and two that we introduced [Exploitation and Combination_XA]) significantly reduced (by 48% to 64%) condition labeling efforts, compared to standard passive (random instance-selection) SVM learning. Furthermore, our new AL methods achieved maximal accuracy using 12% fewer labeled cases than the SVM-Margin AL method. However, because labelers have varying levels of expertise, a major issue associated with learning methods, and AL methods in particular, is how to best to use the labeling provided by a committee of labelers. First, we wanted to know, based on the labelers' learning curves, whether using AL methods (versus standard passive learning methods) has an effect on the Intra-labeler variability (within the learning curve of each labeler) and inter-labeler variability (among the learning curves of different labelers). Then, we wanted to examine the effect of learning (either passively or actively) from the labels created by the majority consensus of a group of labelers. We used our CAESAR-ALE framework for classifying the severity of clinical conditions, the three AL methods and the passive learning method, as mentioned above, to induce the classifications models. We used a dataset of 516 clinical conditions and their severity labeling, represented by features aggregated from the medical records of 1.9 million patients treated at Columbia University Medical Center. We analyzed the variance of the classification performance within (intra-labeler), and especially among (inter-labeler) the classification models that were induced by using the labels provided by seven

  6. Inter-Labeler and Intra-Labeler Variability of Condition Severity Classification Models Using Active and Passive Learning Methods

    Science.gov (United States)

    Nissim, Nir; Shahar, Yuval; Boland, Mary Regina; Tatonetti, Nicholas P; Elovici, Yuval; Hripcsak, George; Moskovitch, Robert

    2018-01-01

    Background and Objectives Labeling instances by domain experts for classification is often time consuming and expensive. To reduce such labeling efforts, we had proposed the application of active learning (AL) methods, introduced our CAESAR-ALE framework for classifying the severity of clinical conditions, and shown its significant reduction of labeling efforts. The use of any of three AL methods (one well known [SVM-Margin], and two that we introduced [Exploitation and Combination_XA]) significantly reduced (by 48% to 64%) condition labeling efforts, compared to standard passive (random instance-selection) SVM learning. Furthermore, our new AL methods achieved maximal accuracy using 12% fewer labeled cases than the SVM-Margin AL method. However, because labelers have varying levels of expertise, a major issue associated with learning methods, and AL methods in particular, is how to best to use the labeling provided by a committee of labelers. First, we wanted to know, based on the labelers’ learning curves, whether using AL methods (versus standard passive learning methods) has an effect on the Intra-labeler variability (within the learning curve of each labeler) and inter-labeler variability (among the learning curves of different labelers). Then, we wanted to examine the effect of learning (either passively or actively) from the labels created by the majority consensus of a group of labelers. Methods We used our CAESAR-ALE framework for classifying the severity of clinical conditions, the three AL methods and the passive learning method, as mentioned above, to induce the classifications models. We used a dataset of 516 clinical conditions and their severity labeling, represented by features aggregated from the medical records of 1.9 million patients treated at Columbia University Medical Center. We analyzed the variance of the classification performance within (intra-labeler), and especially among (inter-labeler) the classification models that were induced by

  7. Site Specific Probable Maximum Precipitation Estimates and Professional Judgement

    Science.gov (United States)

    Hayes, B. D.; Kao, S. C.; Kanney, J. F.; Quinlan, K. R.; DeNeale, S. T.

    2015-12-01

    State and federal regulatory authorities currently rely upon the US National Weather Service Hydrometeorological Reports (HMRs) to determine probable maximum precipitation (PMP) estimates (i.e., rainfall depths and durations) for estimating flooding hazards for relatively broad regions in the US. PMP estimates for the contributing watersheds upstream of vulnerable facilities are used to estimate riverine flooding hazards while site-specific estimates for small water sheds are appropriate for individual facilities such as nuclear power plants. The HMRs are often criticized due to their limitations on basin size, questionable applicability in regions affected by orographic effects, their lack of consist methods, and generally by their age. HMR-51 for generalized PMP estimates for the United States east of the 105th meridian, was published in 1978 and is sometimes perceived as overly conservative. The US Nuclear Regulatory Commission (NRC), is currently reviewing several flood hazard evaluation reports that rely on site specific PMP estimates that have been commercially developed. As such, NRC has recently investigated key areas of expert judgement via a generic audit and one in-depth site specific review as they relate to identifying and quantifying actual and potential storm moisture sources, determining storm transposition limits, and adjusting available moisture during storm transposition. Though much of the approach reviewed was considered a logical extension of HMRs, two key points of expert judgement stood out for further in-depth review. The first relates primarily to small storms and the use of a heuristic for storm representative dew point adjustment developed for the Electric Power Research Institute by North American Weather Consultants in 1993 in order to harmonize historic storms for which only 12 hour dew point data was available with more recent storms in a single database. The second issue relates to the use of climatological averages for spatially

  8. Document clustering methods, document cluster label disambiguation methods, document clustering apparatuses, and articles of manufacture

    Science.gov (United States)

    Sanfilippo, Antonio [Richland, WA; Calapristi, Augustin J [West Richland, WA; Crow, Vernon L [Richland, WA; Hetzler, Elizabeth G [Kennewick, WA; Turner, Alan E [Kennewick, WA

    2009-12-22

    Document clustering methods, document cluster label disambiguation methods, document clustering apparatuses, and articles of manufacture are described. In one aspect, a document clustering method includes providing a document set comprising a plurality of documents, providing a cluster comprising a subset of the documents of the document set, using a plurality of terms of the documents, providing a cluster label indicative of subject matter content of the documents of the cluster, wherein the cluster label comprises a plurality of word senses, and selecting one of the word senses of the cluster label.

  9. Immunoreactivity of 125I-papain labelled by different methods

    International Nuclear Information System (INIS)

    Rauch, P.; Fukal, L.; Kas, J.; Tykva, R.

    1984-01-01

    Three different methods of papain iodination (with chloramine-T, lactoperoxidase and conjugation with Bolton-Hunter reagent) have been compared. The highest yield of 125 I-papain could be obtained using lactoperoxidase which enabled to achieve the highest immunoreactivity. 125 I-papain, labelled this way, is suitable for the radioimmunoassay of papain. (author)

  10. Modern spectrometric methods for the analysis of labelled compounds

    International Nuclear Information System (INIS)

    Kaspersen, F.M.; Funke, C.W.; Wagenaars, G.N.; Jacobs, P.L.

    1988-01-01

    A proper analysis of chemical compounds should give information about the chemical identity (not only the structure but also enantiomeric form), the chemical purity and chemical composition (e.g. giving information about counter-ions, solvents of crystallization). For labelled compounds information is also needed about isotopic purity (defined as the % of isotope present in the compound), the position/distribution of the isotope in the molecule and degree of labelling/specific activity. In the past ten years the possibilities for spectrometric analyses of labelled compounds have increased enormously and this chapter will give an overview of these methods with the exception of (radio)chromatography that will be dealt with in another chapter. (author)

  11. Community Mining Method of Label Propagation Based on Dense Pairs

    Directory of Open Access Journals (Sweden)

    WENG Wei

    2014-03-01

    Full Text Available In recent years, with the popularity of handheld Internet equipments like mobile phones, increasing numbers of people are becoming involved in the virtual social network. Because of its large amount of data and complex structure, the network faces new challenges of community mining. A label propagation algorithm with low time complexity and without prior parameters deals easily with a large networks. This study explored a new method of community mining, based on label propagation with two stages. The first stage involved identifying closely linked nodes according to their local adjacency relations that gave rise to a micro-community. The second stage involved expanding and adjusting this community through a label propagation algorithm (LPA to finally obtain the community structure of the entire social network. This algorithm reduced the number of initial labels and avoided the merging of small communities in general LPAs. Thus, the quality of community discovery was improved, and the linear time complexity of the LPA was maintained.

  12. Development of radio-labeling method for natural juvenile hormone

    International Nuclear Information System (INIS)

    Kurata, Keiji; Shiozuki, Takahiro; Kotaki, Toyomi

    1997-01-01

    The aim of this study was to develop a new method for quantitative determination of juvenile hormone (JH) based on the principle for radioimmuno assay. Using JH-binding protein (JHBP), the discrimination of the L-form of JH from D-form not occurring naturally was attempted to establish a measuring method for JH. First, the corpus allatum, the JH-producing endocrine organ was cultured and both L-form JH and 3 H or 14 C-labelled JH could be obtained easily. There are several homologs of JH and it is necessary to establish the respective labelling methods for the homologues. Since different species of insects produce JH with its specific structure, each homologue could be produced by selecting an appropriate species. The capacity of JH production was compared among five insects. The biosynthetic ability by the corpus allatum from migratory locust was highest among them and 1 μg of labelled JH type 3 could be obtained from the culture with about 50 corpus allata for 3 hrs. Since other materials than JH were also released into the culture medium, establishment of the effective method for isolation and purification of JH is necessary to use it for bioassay. (M.N.)

  13. Innovation and Diffusion of Site-specific Crop Management

    DEFF Research Database (Denmark)

    Pedersen, Søren Marcus; Pedersen, Jørgen Lindgaard

    2006-01-01

    Site-specific crop management or precision farming is a highly complex managementsystem for site-specific input application of lime, fertilizers and pesticides in arable farming. The Global Positioning System (GPS)is the backbone of the system. To conduct precision farming several technical systems...

  14. Innovation and diffusion of site-specific crop management

    DEFF Research Database (Denmark)

    Pedersen, Søren Marcus; Pedersen, Jørgen Lindgaard

    2004-01-01

    Site-specific crop management or precision farming (PF) is a highly complex management system for site-specific input application of lime, fertilizers and pesticides in arable farming. The Global Positioning System (GPS) is the backbone of the system. To conduct PF several technical systems...

  15. Validation of single-sample doubly labeled water method

    International Nuclear Information System (INIS)

    Webster, M.D.; Weathers, W.W.

    1989-01-01

    We have experimentally validated a single-sample variant of the doubly labeled water method for measuring metabolic rate and water turnover in a very small passerine bird, the verdin (Auriparus flaviceps). We measured CO 2 production using the Haldane gravimetric technique and compared these values with estimates derived from isotopic data. Doubly labeled water results based on the one-sample calculations differed from Haldane values by less than 0.5% on average (range -8.3 to 11.2%, n = 9). Water flux computed by the single-sample method differed by -1.5% on average from results for the same birds based on the standard, two-sample technique (range -13.7 to 2.0%, n = 9)

  16. Site-Specific Antibody Functionalization Using Tetrazine-Styrene Cycloaddition.

    Science.gov (United States)

    Umlauf, Benjamin J; Mix, Kalie A; Grosskopf, Vanessa A; Raines, Ronald T; Shusta, Eric V

    2018-05-03

    Biologics, such as antibody-drug conjugates, are becoming mainstream therapeutics. Consequently, methods to functionalize biologics without disrupting their native properties are essential for identifying, characterizing, and translating candidate biologics from the bench to clinical practice. Here, we present a method for site-specific, carboxy-terminal modification of single-chain antibody fragments (scFvs). ScFvs displayed on the surface of yeast were isolated and functionalized by combining intein-mediated expressed protein ligation (EPL) with inverse electron-demand Diels-Alder (IEDDA) cycloaddition using a styrene-tetrazine pair. The high thiol concentration required to trigger EPL can hinder the subsequent chemoselective ligation reactions; therefore, the EPL reaction was used to append styrene to the scFv, limiting tetrazine exposure to damaging thiols. Subsequently, the styrene-functionalized scFv was reacted with tetrazine-conjugated compounds in an IEDDA cycloaddition to generate functionalized scFvs that retain their native binding activity. Rapid functionalization of yeast surface-derived scFv in a site-directed manner could find utility in many downstream laboratory and preclinical applications.

  17. Alternative methods of synthesizing 99Tcm-labelled ciprofloxacin

    International Nuclear Information System (INIS)

    Kumar, V.; Choong, K.K.L.; Evans, S.; Olma, T.R.

    1999-01-01

    Full text: 99 Tc m -labelled ciprofloxacin (Infecton) is a new class of radiopharmaceutical designed for imaging live bacterial infection. We synthesized Infecton by modifying the procedure described by Keith Britton's group (Lancet 1996; 347: 233-235) and reported our findings at the ANZSNM meeting last year. Since the methodology was cumbersome, we investigated simpler alternative ways of labelling ciprofloxacin with 99 Tc m -pertechnetate for routine imaging. There were several limitations in the previously described method: (1) Need to prepare pure ciprofloxacin which was unstable on storage. (2) Synthetic procedure using formimidine sulphinic acid (FSA) was complicated and required boiling step. (3) The radiochemical purity (RCP) of the product was low (45-50%) requiring purification. (4) Biodistribution studies showed a marked uptake by the liver which could interfere with scan interpretation in this region. The results of our present studies showed that Infecton could be prepared by a simple two-step method: (1) Reduce 99 Tc m -pertechnetate with stannous salt (SnCl 2 or Sn-tartrate). (2) Mix with Ciproxin IV-100. The RCP of the product was up to 98%, which obviates the need for further purification. Infecton synthesized by the above method showed avid localization in abscesses induced with Staphylococcus aureus in rats. The biodistribution studies showed that Infecton was renally excreted with minimal accumulation in the liver or other organs

  18. Labelled antibody assays for measuring free triiodothyronine: evaluation and comparison with a labelled analog method

    International Nuclear Information System (INIS)

    Sapin, R.; Gasser, F.; Schlienger, J.L.; Chambron, J.

    1993-01-01

    We evaluated analytically and clinically two new one-step labelled antibody assays for measuring free triiodothyronine (FT3): the first, radiolabelled with 125 I, Amerlex-MAB (MAB) from Kodak diagnostic, and the second, labelled with peroxidase, Enzymum-test FT3 (BM) from Boehringer Mannheim adapted for the Boehringer ES 600 analyzer. The clinical results were compared with those obtained with a radiolabelled analog tracer kit, Amerlex-M (M) from Kodak diagnostic. The latter kit is known to give low FT3 results in sera with low albumin concentrations. Analytical performances of the automated method (BM) were better than those obtained with the manual method (MAB): intra-assay reproducibility (CV<3% vs CV about 5%), inter-assay reproducibility (CV<4% vs CV between 4 and 8%) and mean drift (+1.25% vs -4.3%). The detection limit was low for both kits (<1 pmol/l). In the euthyroid reference group (n = 98) we observed a significant difference between outpatient and hospitalized patient FT3 concentrations as measured with the M kit only. Clinical sensitivity for hyperthyroidism (n = 38) was better for the MAB (92%) than for the BM kit (76%). Specificity in euthyroid L-thyroxine (T4) treated patients (n = 26) was good for both kits (MAB: 92%; BM: 88%) . Hypoalbuminemia (n = 8) decreased FT3 results as follows: M, very significantly; BM, significantly; MAB, only slightly. In patients treated with amiodarone (n = 5), a drug known to lower the metabolic conversion of T4 to T3, the increase of the MAB FT3 results contrasted with the decrease of the BM and M results. In conclusion, results of the two new kits were not strongly influenced by hypoalbuminemia. The MAB results showing lack of decrease in patients with non-thyroidal illness without hypoalbuminemia and in amiodarone-treated patients were unexpected

  19. Efficient method of enzymatic synthesis of nucleosides labelled with 14C and 3H

    International Nuclear Information System (INIS)

    Nejedly, Z.; Filip, J.

    1988-01-01

    The method is presented of enzymatic synthesis of nucleosides labelled with 14 C or 3 H either uniformly or specifically in the base or the deoxyribosyl or ribosyl moiety. The method is based on the ribosylation or deoxyribosylation of the nucleic acid bases (non-labelled or labelled with 14 C or 3 H) by the catalytic effect of enzymes occurring in the supernatant fractions of non-purified homogenates of Escherichia coli B. bacteria. The non-labelled and labelled nucleosides are used as donors of ribosyl or deoxyribosyl groups. The HPLC method is used for separating labelled nucleosides. The radiochemical purity of the labelled nucleosides is higher than 98%, molar activity ranges from 9.2 to 18.5 GBq.mmol -1 ( 14 C-labelled compounds) and from 0.6 to 1.9 TBq.mmol -1 (3H-labelled compounds). (author). 4 figs., 8 refs

  20. Micro-tattoo guided OCT imaging of site specific inflammation

    Science.gov (United States)

    Phillips, Kevin G.; Choudhury, Niloy; Samatham, Ravikant V.; Singh, Harvinder; Jacques, Steven L.

    2010-02-01

    Epithelial biologists studying human skin diseases such as cancer formation and psoriasis commonly utilize mouse models to characterize the interplay among cells and intracellular signal transduction pathways that result in programmed changes in gene expression and cellular behaviors. The information obtained from animal models is useful only when phenotypic presentations of disease recapitulate those observed in humans. Excision of tissues followed by histochemical analysis is currently the primary means of establishing the morphological presentation. Non invasive imaging of animal models provides an alternate means to characterize tissue morphology associated with the disease of interest in vivo. While useful, the ability to perform in vivo imaging at different time points in the same tissue location has been a challenge. This information is key to understanding site specific changes as the imaged tissue can now be extracted and analyzed for mRNA expression. We present a method employing a micro-tattoo to guide optical coherence tomography (OCT) imaging of ultraviolet induced inflammation over time in the same tissue locations.

  1. Assay for vitamin B12 absorption and method of making labeled vitamin B12

    Science.gov (United States)

    Anderson, Peter J [Davis, CA; Dueker, Stephen [Davis, CA; Miller, Joshua [Davis, CA; Green, Ralph [Elmacero, CA; Roth, John [Davis, CA; Carkeet, Colleen [Silver Spring, MD; Buchholz,; Bruce, A [Orinda, CA

    2012-06-19

    The invention provides methods for labeling vitamin B12 with .sup.14C, .sup.13C, tritium, and deuterium. When radioisotopes are used, the invention provides for methods of labeling B12 with high specific activity. The invention also provides labeled vitamin B12 compositions made in accordance with the invention.

  2. Labeling and stability of radiolabeled antibody fragments by a direct 99mTc-labeling method

    International Nuclear Information System (INIS)

    Pak, K.Y.; Nedelman, M.A.; Tam, S.H.; Wilson, E.; Daddona, P.E.

    1992-01-01

    The in vitro labeling and stability of 99m Tc-labeled antibody Fab' fragments prepared by a direct labeling technique were evaluated. Eight antibody fragments derived from murine IgG1 (N = 5), IgG2a (N = 2) and IgG3 (N = 1) isotypes were labeled with a preformed 99m Tc-D-glucarate complex. No loss of radioactivity incorporation was observed for all the 99m Tc-labeled antibody fragments after 24 h incubation at 37 o C. 99m Tc-labeled antibody fragments (IgG1, N = 2; IgG2a, n = 2; IgG3, N = 1) were stable upon challenge with DTPA, EDTA or acidic pH. Using the affinity chromatography technique, two of the 99m Tc-labeled antibody fragments displayed no loss of immunoreactivity after prolonged incubation in phosphate buffer up to 24 h at 37 o C. Bonding between 99m Tc and antibody fragments was elucidated by challenging with a diamide ditholate (N 2 S 2 ) compound. The Fab' with IgG2a isotype displayed tighter binding to 99m Tc in comparison to Fab' from IgG1 and IgG3 isotype in N 2 S 2 challenge and incubation with human plasma. The in vivo biodistribution of five 99m Tc-labeled fragments were evaluated in normal mice. (Author)

  3. Prediction of site specific ground motion for large earthquake

    International Nuclear Information System (INIS)

    Kamae, Katsuhiro; Irikura, Kojiro; Fukuchi, Yasunaga.

    1990-01-01

    In this paper, we apply the semi-empirical synthesis method by IRIKURA (1983, 1986) to the estimation of site specific ground motion using accelerograms observed at Kumatori in Osaka prefecture. Target earthquakes used here are a comparatively distant earthquake (Δ=95 km, M=5.6) caused by the YAMASAKI fault and a near earthquake (Δ=27 km, M=5.6). The results obtained are as follows. 1) The accelerograms from the distant earthquake (M=5.6) are synthesized using the aftershock records (M=4.3) for 1983 YAMASAKI fault earthquake whose source parameters have been obtained by other authors from the hypocentral distribution of the aftershocks. The resultant synthetic motions show a good agreement with the observed ones. 2) The synthesis for a near earthquake (M=5.6, we call this target earthquake) are made using a small earthquake which occurred in the neighborhood of the target earthquake. Here, we apply two methods for giving the parameters for synthesis. One method is to use the parameters of YAMASAKI fault earthquake which has the same magnitude as the target earthquake, and the other is to use the parameters obtained from several existing empirical formulas. The resultant synthetic motion with the former parameters shows a good agreement with the observed one, but that with the latter does not. 3) We estimate the source parameters from the source spectra of several earthquakes which have been observed in this site. Consequently we find that the small earthquakes (M<4) as Green's functions should be carefully used because the stress drops are not constant. 4) We propose that we should designate not only the magnitudes but also seismic moments of the target earthquake and the small earthquake. (J.P.N.)

  4. IL-2 labeled with 99mTechnetium by an indirect method

    International Nuclear Information System (INIS)

    Bocco, R.; Obenaus, Esteban; Rabiller, Graciela; Castiglia, Silvia G. de

    2003-01-01

    IL-2 and the other cytokines labeled with 99m Tc are an interesting option to early diagnosis of autoimmune diseases and monitoring with nuclear medicine images. The aim of this study was to obtain by indirect method IL-2 labeled with 99m Technetium using Benzoyl MAG3 chelating agent, for in vivo diagnosis of lymphocytic infiltration. IL-2 is a small, relatively fragile protein, and it is essential to retain its receptor binding capacity after labeling. Two different methods of labeling have been proven: Pre-conjugation labeling method: we used NHS- Hynic as a chelator agent and labeled this conjugated protein with 99m Tc using tricine as coligand. Albumin was used as a model for the conjugation and labeling steps. The albumin was labeled by this method with a good labeling efficiency but the IL-2 protein could not be labeled with this approach. Post- conjugation labeling method: we used the bifunctional chelating agent, benzoyl MAG3, this ligand is first labeled with 99m Tc and then is conjugated to the protein. The N 3 S Logan complex was incubated for 30 minutes and then measured by RP- HPLC. An active ester of the labeled Logan was formed and was incubated with Il-2 at room temperature and basic pH to promote the conjugation between the active ester and the protein. The labeling efficiency was determined by RP-HPLC using a C 18 column. The albumin protein was also labeled by this method and the radiochemical purity was measured by RP- HPLC using a GPC column and the labeling efficiency was 80%. The next objectives are to explore different strategies for purifying the 99m Tc-IL-2 and to evaluate the capacity of IL-2 to bind to its receptor after labeling. (author)

  5. Some methods for labelling organic compounds by deuterium

    International Nuclear Information System (INIS)

    Moustapha, C.

    1988-01-01

    The rapid growth of knowledge in the fields of biochemistry, physiology, and molecular biology reflects to a considerable degree the utilization of stable isotopes (specially deuterium) in the study of chemical reactions and fragmentation mechanisms in mass spectrometry, as well as in the pharmacological and biological studies. Organic compounds maybe labelled by deuterium through classic organic reactions by using special deuterated solvents and reagents. This article discusses some reactions, with examples on how to prepare labelled compounds with high isotopic purety. These reactions are: exchange reactions in acid and alkaline media (the exchange in the chromatographic column in liquid and gas phases, the exchange in homogenous medium), reduction reactions of functional groups as well as saturation of the double bounds by deuterium using hydrogenation catalystes, electrochemical reactions using KOLBE, and photochemical reactions. This article also deals with spectroscopic properties of deuterium and the methods which are used to identify its compounds such as infrared, nuclear magnetic resonance, and mass spectroscopy. 37 refs., 2 figs

  6. Evaluation of potential water conservation using site-specific irrigation

    Science.gov (United States)

    With the advent of site-specific variable-rate irrigation (VRI) systems, irrigation can be spatially managed within sub-field-sized zones. Spatial irrigation management can optimize spatial water use efficiency and may conserve water. Spatial VRI systems are currently being managed by consultants ...

  7. 77 FR 22772 - Environmental Management Site-Specific Advisory Board

    Science.gov (United States)

    2012-04-17

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board AGENCY: Office of Environmental Management, Department of Energy. ACTION: Notice of renewal. SUMMARY: Pursuant to Section 14(a)(2... Secretariat, General Services Administration, notice is hereby given that the Environmental Management Site...

  8. Determination of site-specific glycan heterogeneity on glycoproteins

    DEFF Research Database (Denmark)

    Kolarich, Daniel; Jensen, Pia Hønnerup; Altmann, Friedrich

    2012-01-01

    and the determination of site-specific glycan heterogeneity. The described workflow takes approximately 3-5 d, including sample preparation and data analysis. The data obtained from analyzing released glycans of rHuEPO and IgG, described in the second protocol of this series (10.1038/nprot.2012.063), provide...

  9. Precision agriculture - from mapping to site-specific application

    DEFF Research Database (Denmark)

    Pedersen, Søren Marcus; Lind, Kim Martin Hjorth

    2017-01-01

    of each chapter in the book. Each chapter address a different topic starting with an overview of technologies that are currently available, followed by specific Variable-Rate Technologies such as VRT fertilizer application, VRT pesticide application, site-specific irrigation management, Auto...

  10. Development of method of tritium labeling of pharmacological preparate of drotaverine hydrochloride (NOSPA)

    International Nuclear Information System (INIS)

    Kim, A.A.; Djuraeva, G.T.; Shukurov, B.V.

    2004-01-01

    Full text: The method for tritium labeling of pharmacological preparate of drotaverine hydrochloride (no spa) was developed. Drotaverine hydrochloride was labeled by thermally activated tritium in apparatus for tritium labeling. The optimum regime of labeling was selected. The system of purification of tritium labeled drotaverine hydrochloride by thin layer chromatography (TLC) has been developed. The TLC system of purification of tritium labeled drotaverine hydrochloride was developed. Tritium labeled preparation of drotaverine hydrochloride was purified by TLC on silicagel in system isopropanol: ammonia: water (8:1:1). We found appearance of additional fractions in tritium labeled preparation of drotaverine hydrochloride that testifies to partial transformation of drotaverine hydrochloride during procedure of labeling. Application of TLC for purification of tritium labeled preparation allows to purify completely drotaverine hydrochloride of by-products. The output of purified tritium labeled preparation of drotaverine hydrochloride was about 25 %. The received preparation had specific radioactivity - 3,2 MBq/mg, radiochemical purity of a preparation was 95 %. TLC purification seems inexpensive, fast and suitable for purification of tritium-labeled drotaverine hydrochloride. Thus developed method allows obtain tritium labeled preparation of drotaverine hydrochloride (no - spa), suitable for medical and biologic researches

  11. ETAC reagents: A new class of sulfhydryl site-specific radiolabelling probes for antibodies

    International Nuclear Information System (INIS)

    del Rosario, R.B.; Brocchini, S.J.; Baron, L.A.; Smith, R.H.; Lawton, R.G.; Wahl, R.L.

    1990-01-01

    A new class of bis-alkylating Michael reagents, equilibrium transfer crosslink reagents, 'ETAC', which combine the techniques of crosslinking with tethering have been synthesized. Following a succession of Michael and retro-Michael additions and elimination of the arylsulfone groups, reduced heavy-heavy and heavy-light disulfide links of an anti-ovarian IgG2a monoclonal antibody, 5G6.4, were site-specifically re-annealed via a 3-carbon bridge having a tether branch containing a designated label

  12. 16 CFR 301.27 - Label and method of affixing.

    Science.gov (United States)

    2010-01-01

    ... all times during the marketing of a fur product the required label shall have a minimum dimension of one and three-fourths (13/4) inches by two and three-fourths (23/4) inches (4.5 cm × 7 cm). Such label...

  13. Improving Site-Specific Radiological Performance Assessments - 13431

    International Nuclear Information System (INIS)

    Tauxe, John; Black, Paul; Catlett, Kate; Lee, Robert; Perona, Ralph; Stockton, Tom; Sully, Mike

    2013-01-01

    An improved approach is presented for conducting complete and defensible radiological site-specific performance assessments (PAs) to support radioactive waste disposal decisions. The basic tenets of PA were initiated some thirty years ago, focusing on geologic disposals and evaluating compliance with regulations. Some of these regulations were inherently probabilistic (i.e., addressing uncertainty in a quantitative fashion), such as the containment requirements of the U.S. Environmental Protection Agency's (EPA's) 40 CFR 191, Environmental Radiation Protection Standards for Management and Disposal of Spent Nuclear Fuel, High-Level and Transuranic Radioactive Wastes, Chap. 191.13 [1]. Methods of analysis were developed to meet those requirements, but at their core early PAs used 'conservative' parameter values and modeling approaches. This limited the utility of such PAs to compliance evaluation, and did little to inform decisions about optimizing disposal, closure and long-term monitoring and maintenance, or, in general, maintaining doses 'as low as reasonably achievable' (ALARA). This basic approach to PA development in the United States was employed essentially unchanged through the end of the 20. century, principally by the U.S. Department of Energy (DOE). Performance assessments developed in support of private radioactive waste disposal operations, regulated by the U.S. Nuclear Regulatory Commission (NRC) and its agreement states, were typically not as sophisticated. Discussion of new approaches to PA is timely, since at the time of this writing, the DOE is in the midst of revising its Order 435.1, Radioactive Waste Management [2], and the NRC is revising 10 CFR 61, Licensing Requirements for Land Disposal of Radioactive Waste [3]. Over the previous decade, theoretical developments and improved computational technology have provided the foundation for integrating decision analysis (DA) concepts and objective-focused thinking, plus a Bayesian approach to

  14. Improving Site-Specific Radiological Performance Assessments - 13431

    Energy Technology Data Exchange (ETDEWEB)

    Tauxe, John; Black, Paul; Catlett, Kate; Lee, Robert; Perona, Ralph; Stockton, Tom; Sully, Mike [Neptune and Company, Inc., Los Alamos, New Mexico 87544 (United States)

    2013-07-01

    An improved approach is presented for conducting complete and defensible radiological site-specific performance assessments (PAs) to support radioactive waste disposal decisions. The basic tenets of PA were initiated some thirty years ago, focusing on geologic disposals and evaluating compliance with regulations. Some of these regulations were inherently probabilistic (i.e., addressing uncertainty in a quantitative fashion), such as the containment requirements of the U.S. Environmental Protection Agency's (EPA's) 40 CFR 191, Environmental Radiation Protection Standards for Management and Disposal of Spent Nuclear Fuel, High-Level and Transuranic Radioactive Wastes, Chap. 191.13 [1]. Methods of analysis were developed to meet those requirements, but at their core early PAs used 'conservative' parameter values and modeling approaches. This limited the utility of such PAs to compliance evaluation, and did little to inform decisions about optimizing disposal, closure and long-term monitoring and maintenance, or, in general, maintaining doses 'as low as reasonably achievable' (ALARA). This basic approach to PA development in the United States was employed essentially unchanged through the end of the 20. century, principally by the U.S. Department of Energy (DOE). Performance assessments developed in support of private radioactive waste disposal operations, regulated by the U.S. Nuclear Regulatory Commission (NRC) and its agreement states, were typically not as sophisticated. Discussion of new approaches to PA is timely, since at the time of this writing, the DOE is in the midst of revising its Order 435.1, Radioactive Waste Management [2], and the NRC is revising 10 CFR 61, Licensing Requirements for Land Disposal of Radioactive Waste [3]. Over the previous decade, theoretical developments and improved computational technology have provided the foundation for integrating decision analysis (DA) concepts and objective-focused thinking, plus

  15. Recent advances in covalent, site-specific protein immobilization [version 1; referees

    DEFF Research Database (Denmark)

    Meldal, Morten Peter; Schoffelen, Sanne

    2016-01-01

    The properties of biosensors, biomedical implants, and other materials based on immobilized proteins greatly depend on the method employed to couple the protein molecules to their solid support. Covalent, site-specific immobilization strategies are robust and can provide the level of control...

  16. Site specific study for possible ongoing salt dome movement

    International Nuclear Information System (INIS)

    Thoms, R.L.; Manning, T.A.; Paille, L.K.; Gehle, R.M.

    1977-01-01

    U.S. Gulf Coast salt domes, among other geologic structures, currently are being considered for storage of commercial radioactive wastes. A major concern with dome storage of long lived radioactive wastes lies with the possible tectonic movement of the host dome. Any ongoing movement of a salt dome can be monitored with a site specific complementary system of field instrumentation and finite element modelling. Field instrumentation and accompanying finite element analyses for a study dome in northwest Louisiana are described. Site specific data and early experience associated with tiltmeters over the dome are presented. Also, recommendations are made for modifications and extensions of the field instrumentation and finite element modelling appropriate to the specific site under study

  17. Pinellas Plant FY1990 site specific implementation plan

    International Nuclear Information System (INIS)

    Klein, R.D.

    1990-02-01

    This Site Specific Implementation Plan describes the Corrective Action, Environmental Restoration, and Waste Management activities to be performed at the Pinellas Plant in FY1990 (October 1, 1989 to September 30, 1989). These FY1990 activities are described in the Pinellas Plant FY1991--95 Five-Year Plan. The information used to prepare this plan reflects the best estimate of the project scope, schedules, regulatory, and funding requirements at the time of plan preparation. The Environmental Restoration/Waste Management Five-Year Plan is a dynamic document and will be modified each year; the Site Specific Implementation Plan will, in turn, be modified each year to reflect new findings, information, and knowledge of the various projects. 4 figs., 11 tabs

  18. Site-specific design optimization of wind turbines

    DEFF Research Database (Denmark)

    Fuglsang, P.; Bak, C.; Schepers, J.G.

    2002-01-01

    This article reports results from a European project, where site characteristics were incorporated into the design process of wind turbines, to enable site-specific design. Two wind turbines of different concept were investigated at six different sites comprising normal flat terrain, offshore...... and complex terrain wind farms. Design tools based on numerical optimization and aeroelastic calculations were combined with a cost model to allow optimization for minimum cost of energy. Different scenarios were optimized ranging from modifications of selected individual components to the complete design...... of a new wind turbine. Both annual energy yield and design-determining loads depended on site characteristics, and this represented a potential for site-specific design. The maximum variation in annual energy yield was 37% and the maximum variation in blade root fatigue loads was 62%. Optimized site...

  19. Intruder scenarios for site-specific waste classification

    International Nuclear Information System (INIS)

    Kennedy, W.E. Jr.

    1988-01-01

    The US Department of Energy (DOE) is currently revising its low-level radioactive waste (LLW) management requirements and guidelines for waste generated at its facilities that support defense missions. Specifically, draft DOE 5820.2A, Chapter 3, describes the purpose, policy, and requirements necessary for the management of defense LLW. The draft DOE policy calls for DOE LLW operations to be managed to protect the health and safety of the public, preserve the environment, and ensure that no remedial action will be necessary after termination of operations. The requirements and guidelines apply to radioactive wastes but are also intended to apply to mixed hazardous and radioactive wastes as defined in draft DOE 5400.5, Hazardous and Radioactive Mixed Waste. The basic approach used by DOE is to establish overall performance objectives in terms of ground-water protection and public radiation dose limits and to require site-specific performance assessments to determine compliance. As a result of these performance assessments, each site shall develop waste acceptance criteria that define the allowable quantities and concentrations of specific radioisotopes. Additional limitations on waste disposal design, waste form, and waste treatment shall also be developed on a site-specific basis. As a key step in the site-specific performance assessments, an evaluation must be conducted of potential radiation doses to intruders who may inadvertently move onto a closed DOE LLW disposal site after loss of institutional controls must be conducted. This paper describes the types of intruder scenarios that should be considered when performing this step of the site-specific performance assessment

  20. Site-specific Probabilistic Analysis of DCGLs Using RESRAD Code

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Jeongju; Yoon, Suk Bon; Sohn, Wook [KHNP CRI, Daejeon (Korea, Republic of)

    2016-10-15

    In general, DCGLs can be conservative (screening DCGL) if they do not take into account site specific factors. Use of such conservative DCGLs can lead to additional remediation that would not be required if the effort was made to develop site-specific DCGLs. Therefore, the objective of this work is to provide an example on the use of the RESRAD 6.0 probabilistic (site-specific) dose analysis to compare with the screening DCGL. Site release regulations state that a site will be considered acceptable for unrestricted use if the residual radioactivity that is distinguishable from background radiation results in a Total Effective Dose Equivalent (TEDE) to an average member of the critical group of less than the site release criteria, for example 0.25 mSv per year in U.S. Utilities use computer dose modeling codes to establish an acceptable level of contamination, the derived concentration guideline level (DCGL) that will meet this regulatory limit. Since the DCGL value is the principal measure of residual radioactivity, it is critical to understand the technical basis of these dose modeling codes. The objective this work was to provide example on nuclear power plant decommissioning dose analysis in a probabilistic analysis framework. The focus was on the demonstration of regulatory compliance for surface soil contamination using the RESRAD 6.0 code. Both the screening and site-specific probabilistic dose analysis methodologies were examined. Example analyses performed with the screening probabilistic dose analysis confirmed the conservatism of the NRC screening values and indicated the effectiveness of probabilistic dose analysis in reducing the conservatism in DCGL derivation.

  1. Site specific atomic polarizabilities in endohedral fullerenes and carbon onions

    Energy Technology Data Exchange (ETDEWEB)

    Zope, Rajendra R., E-mail: rzope@utep.edu; Baruah, Tunna [Department of Physics, The University of Texas at El Paso, El Paso, Texas 79958 (United States); Computational Science Program, The University of Texas at El Paso, El Paso, Texas 79958 (United States); Bhusal, Shusil; Basurto, Luis [Department of Physics, The University of Texas at El Paso, El Paso, Texas 79958 (United States); Jackson, Koblar [Physics Department and Science of Advanced Materials Ph.D. Program, Central Michigan University, Mt. Pleasant, Michigan 48859 (United States)

    2015-08-28

    We investigate the polarizability of trimetallic nitride endohedral fullerenes by partitioning the total polarizability into site specific components. This analysis indicates that the polarizability of the endohedral fullerene is essentially due to the outer fullerene cage and has insignificant contribution from the encapsulated unit. Thus, the outer fullerene cages effectively shield the encapsulated clusters and behave like Faraday cages. The polarizability of endohedral fullerenes is slightly smaller than the polarizability of the corresponding bare carbon fullerenes. The application of the site specific polarizabilities to C{sub 60}@C{sub 240} and C{sub 60}@C{sub 180} onions shows that, compared to the polarizability of isolated C{sub 60} fullerene, the encapsulation of the C{sub 60} in C{sub 240} and C{sub 180} fullerenes reduces its polarizability by 75% and 83%, respectively. The differences in the polarizability of C{sub 60} in the two onions is a result of differences in the bonding (intershell electron transfer), fullerene shell relaxations, and intershell separations. The site specific analysis further shows that the outer atoms in a fullerene shell contribute most to the fullerene polarizability.

  2. Site specific atomic polarizabilities in endohedral fullerenes and carbon onions

    International Nuclear Information System (INIS)

    Zope, Rajendra R.; Baruah, Tunna; Bhusal, Shusil; Basurto, Luis; Jackson, Koblar

    2015-01-01

    We investigate the polarizability of trimetallic nitride endohedral fullerenes by partitioning the total polarizability into site specific components. This analysis indicates that the polarizability of the endohedral fullerene is essentially due to the outer fullerene cage and has insignificant contribution from the encapsulated unit. Thus, the outer fullerene cages effectively shield the encapsulated clusters and behave like Faraday cages. The polarizability of endohedral fullerenes is slightly smaller than the polarizability of the corresponding bare carbon fullerenes. The application of the site specific polarizabilities to C 60 @C 240 and C 60 @C 180 onions shows that, compared to the polarizability of isolated C 60 fullerene, the encapsulation of the C 60 in C 240 and C 180 fullerenes reduces its polarizability by 75% and 83%, respectively. The differences in the polarizability of C 60 in the two onions is a result of differences in the bonding (intershell electron transfer), fullerene shell relaxations, and intershell separations. The site specific analysis further shows that the outer atoms in a fullerene shell contribute most to the fullerene polarizability

  3. Site specific atomic polarizabilities in endohedral fullerenes and carbon onions

    Science.gov (United States)

    Zope, Rajendra R.; Bhusal, Shusil; Basurto, Luis; Baruah, Tunna; Jackson, Koblar

    2015-08-01

    We investigate the polarizability of trimetallic nitride endohedral fullerenes by partitioning the total polarizability into site specific components. This analysis indicates that the polarizability of the endohedral fullerene is essentially due to the outer fullerene cage and has insignificant contribution from the encapsulated unit. Thus, the outer fullerene cages effectively shield the encapsulated clusters and behave like Faraday cages. The polarizability of endohedral fullerenes is slightly smaller than the polarizability of the corresponding bare carbon fullerenes. The application of the site specific polarizabilities to C60@C240 and C60@C180 onions shows that, compared to the polarizability of isolated C60 fullerene, the encapsulation of the C60 in C240 and C180 fullerenes reduces its polarizability by 75% and 83%, respectively. The differences in the polarizability of C60 in the two onions is a result of differences in the bonding (intershell electron transfer), fullerene shell relaxations, and intershell separations. The site specific analysis further shows that the outer atoms in a fullerene shell contribute most to the fullerene polarizability.

  4. A microautoradiographic method permitting the study of 99m-technetium labelled leukocytes

    International Nuclear Information System (INIS)

    Colas-Linhart, N.; Perianin, A.; Petiet, A.; Bretillon, A.; Bok, B.

    1985-01-01

    Cell migration studies are very important in inflammatory phenomena. Methods currently used are not quantitative and have been subject to much controversy. Homogeneity of sup(99m)Tc leukocyte labelling was verified by a microautoradiographic method (MAR), which was performed in our laboratory. This method was used in cell migration studies to verify if the migrating cells were indeed the labelled cells [fr

  5. Labelling of blood cells with radioactive indium-201: method, results, indications

    International Nuclear Information System (INIS)

    Ducassou, D.; Brendel, A.; Nouel, J.P.

    1978-01-01

    A modification of the method of Thakur et al. for labelling polynuclear cells with 8-hydroxyquinolein-indium-complexe utilising the water soluble sulfate of the substance was applied. The labelling procedure gave a yield over 98% with erthrocytes and over 80% with platelets and polynuclear cells using at least 1 x 10 8 plasma free cells. Functional capacity of the labelled cells remained unaltered. Injection double labelled ( 111 In, 51 Cr) red cells correlation of values for the red cell volume amounted to r = 0,98 (n=20); red cell life-spane measurements gave comparable results in 5 patients. After injecting labelled platelets a life-spane between 6,5 and 11 days was measured. Scintigraphic visualisation of pulmonary embolism was obtained 30 minutes after injecting labelled platelets. Injection of labelled polynuclear cells allows life-spane measurements as well as detection of abscesses. (author)

  6. The fluorodediazonation - a method for n.c.a.-18F-labelling of aromatic substrates

    International Nuclear Information System (INIS)

    Zwernemann, O.

    1991-06-01

    For the positron emission tomography (PET) applications, radiopharmaceuticals are required that are labelled with short-lived positron emitters. Fluorine-18 has become the leading radionuclide used for PET, due to its favourable physical properties. However, the labelling of aromatic substances with fluorine-18 with the methods available presents problems not encountered with aliphatic compounds. The decomposition of aromatic diazonium salts opens up feasible ways of preparing a broad range of labelled compounds. The dissertation investigated the possibilities of labelling with fluorine-18 by way of dediazonation on the standard substrate p-Toluidyl diazonium ion. The results reported show that the method of fluorodediazonation is an interesting further method for F-18 labelling of aromatic substrates in addition to the hitherto applied techniques. It allows carrier-free labelling of a large group of substances which cannot be fluorinated via direct nucleophilicity. (BBR) [de

  7. A novel dual-isotope labelling method for distinguishing between soil sources of N2O

    NARCIS (Netherlands)

    Wrage, N.; Groenigen, van J.W.; Oenema, O.; Baggs, E.M.

    2005-01-01

    We present a novel O-18-N-15-enrichment method for the distinction between nitrous oxide (N2O) from nitrification, nitrifier denitrification and denitrification based on a method with single- and double-N-15-labelled ammonium nitrate. We added a new treatment with O-18-labelled water to quantify N2O

  8. Statistical methods for quantitative mass spectrometry proteomic experiments with labeling

    Directory of Open Access Journals (Sweden)

    Oberg Ann L

    2012-11-01

    Full Text Available Abstract Mass Spectrometry utilizing labeling allows multiple specimens to be subjected to mass spectrometry simultaneously. As a result, between-experiment variability is reduced. Here we describe use of fundamental concepts of statistical experimental design in the labeling framework in order to minimize variability and avoid biases. We demonstrate how to export data in the format that is most efficient for statistical analysis. We demonstrate how to assess the need for normalization, perform normalization, and check whether it worked. We describe how to build a model explaining the observed values and test for differential protein abundance along with descriptive statistics and measures of reliability of the findings. Concepts are illustrated through the use of three case studies utilizing the iTRAQ 4-plex labeling protocol.

  9. Statistical methods for quantitative mass spectrometry proteomic experiments with labeling.

    Science.gov (United States)

    Oberg, Ann L; Mahoney, Douglas W

    2012-01-01

    Mass Spectrometry utilizing labeling allows multiple specimens to be subjected to mass spectrometry simultaneously. As a result, between-experiment variability is reduced. Here we describe use of fundamental concepts of statistical experimental design in the labeling framework in order to minimize variability and avoid biases. We demonstrate how to export data in the format that is most efficient for statistical analysis. We demonstrate how to assess the need for normalization, perform normalization, and check whether it worked. We describe how to build a model explaining the observed values and test for differential protein abundance along with descriptive statistics and measures of reliability of the findings. Concepts are illustrated through the use of three case studies utilizing the iTRAQ 4-plex labeling protocol.

  10. Simple, rapid method for the preparation of isotopically labeled formaldehyde

    Science.gov (United States)

    Hooker, Jacob Matthew [Port Jefferson, NY; Schonberger, Matthias [Mains, DE; Schieferstein, Hanno [Aabergen, DE; Fowler, Joanna S [Bellport, NY

    2011-10-04

    Isotopically labeled formaldehyde (*C.sup..sctn.H.sub.2O) is prepared from labeled methyl iodide (*C.sup..sctn.H.sub.3I) by reaction with an oxygen nucleophile having a pendant leaving group. The mild and efficient reaction conditions result in good yields of *C.sup..sctn.H.sub.2O with little or no *C isotopic dilution. The simple, efficient production of .sup.11CH.sub.2O is described. The use of the .sup.11CH.sub.2O for the formation of positron emission tomography tracer compounds is described. The reaction can be incorporated into automated equipment available to radiochemistry laboratories. The isotopically labeled formaldehyde can be used in a variety of reactions to provide radiotracer compounds for imaging studies as well as for scintillation counting and autoradiography.

  11. Site specific modification of the human plasma proteome by methylglyoxal

    International Nuclear Information System (INIS)

    Kimzey, Michael J.; Kinsky, Owen R.; Yassine, Hussein N.; Tsaprailis, George; Stump, Craig S.; Monks, Terrence J.; Lau, Serrine S.

    2015-01-01

    Increasing evidence identifies dicarbonyl stress from reactive glucose metabolites, such as methylglyoxal (MG), as a major pathogenic link between hyperglycemia and complications of diabetes. MG covalently modifies arginine residues, yet the site specificity of this modification has not been thoroughly investigated. Sites of MG adduction in the plasma proteome were identified using LC–MS/MS analysis in vitro following incubation of plasma proteins with MG. Treatment of plasma proteins with MG yielded 14 putative MG hotspots from five plasma proteins (albumin [nine hotspots], serotransferrin, haptoglobin [2 hotspots], hemopexin, and Ig lambda-2 chain C regions). The search results revealed two versions of MG-arginine modification, dihydroxyimidazolidine (R + 72) and hydroimidazolone (R + 54) adducts. One of the sites identified was R257 in human serum albumin, which is a critical residue located in drug binding site I. This site was validated as a target for MG modification by a fluorescent probe displacement assay, which revealed significant drug dissociation at 300 μM MG from a prodan–HSA complex (75 μM). Moreover, twelve human plasma samples (six male, six female, with two type 2 diabetic subjects from both genders) were analyzed using multiple reaction monitoring (MRM) tandem mass spectrometry and revealed the presence of the MG-modified albumin R257 peptide. These data provide insights into the nature of the site-specificity of MG modification of arginine, which may be useful for therapeutic treatments that aim to prevent MG-mediated adverse responses in patients. - Highlights: • Methylglyoxal (MG) selectively modifies arginine sites in human plasma proteome. • Dihydroxyimidazolidine and hydroimidazolone adducts on serum albumin identified • MG modification on albumin R257 associated with loss of drug site I binding capacity • MRM-tandem mass spectrometry enables sensitive detection of albumin MG-R257. • Site-specific MG modification may

  12. Site specific modification of the human plasma proteome by methylglyoxal

    Energy Technology Data Exchange (ETDEWEB)

    Kimzey, Michael J.; Kinsky, Owen R. [Southwest Environmental Health Sciences Center, Department of Pharmacology & Toxicology, College of Pharmacy, The University of Arizona, Tucson, AZ 85721 (United States); Yassine, Hussein N. [Department of Medicine, The University of Arizona, Tucson, AZ 85721 (United States); Tsaprailis, George [Southwest Environmental Health Sciences Center, Department of Pharmacology & Toxicology, College of Pharmacy, The University of Arizona, Tucson, AZ 85721 (United States); Stump, Craig S. [Department of Medicine, The University of Arizona, Tucson, AZ 85721 (United States); Southern Arizona VA Health Care System, Tucson, AZ 85723 (United States); Monks, Terrence J. [Southwest Environmental Health Sciences Center, Department of Pharmacology & Toxicology, College of Pharmacy, The University of Arizona, Tucson, AZ 85721 (United States); Lau, Serrine S., E-mail: lau@pharmacy.arizona.edu [Southwest Environmental Health Sciences Center, Department of Pharmacology & Toxicology, College of Pharmacy, The University of Arizona, Tucson, AZ 85721 (United States)

    2015-12-01

    Increasing evidence identifies dicarbonyl stress from reactive glucose metabolites, such as methylglyoxal (MG), as a major pathogenic link between hyperglycemia and complications of diabetes. MG covalently modifies arginine residues, yet the site specificity of this modification has not been thoroughly investigated. Sites of MG adduction in the plasma proteome were identified using LC–MS/MS analysis in vitro following incubation of plasma proteins with MG. Treatment of plasma proteins with MG yielded 14 putative MG hotspots from five plasma proteins (albumin [nine hotspots], serotransferrin, haptoglobin [2 hotspots], hemopexin, and Ig lambda-2 chain C regions). The search results revealed two versions of MG-arginine modification, dihydroxyimidazolidine (R + 72) and hydroimidazolone (R + 54) adducts. One of the sites identified was R257 in human serum albumin, which is a critical residue located in drug binding site I. This site was validated as a target for MG modification by a fluorescent probe displacement assay, which revealed significant drug dissociation at 300 μM MG from a prodan–HSA complex (75 μM). Moreover, twelve human plasma samples (six male, six female, with two type 2 diabetic subjects from both genders) were analyzed using multiple reaction monitoring (MRM) tandem mass spectrometry and revealed the presence of the MG-modified albumin R257 peptide. These data provide insights into the nature of the site-specificity of MG modification of arginine, which may be useful for therapeutic treatments that aim to prevent MG-mediated adverse responses in patients. - Highlights: • Methylglyoxal (MG) selectively modifies arginine sites in human plasma proteome. • Dihydroxyimidazolidine and hydroimidazolone adducts on serum albumin identified • MG modification on albumin R257 associated with loss of drug site I binding capacity • MRM-tandem mass spectrometry enables sensitive detection of albumin MG-R257. • Site-specific MG modification may

  13. Site-Specific Covalent Conjugation of Modified mRNA by tRNA Guanine Transglycosylase.

    Science.gov (United States)

    Ehret, Fabian; Zhou, Cun Yu; Alexander, Seth C; Zhang, Dongyang; Devaraj, Neal K

    2018-03-05

    Modified mRNA (mod-mRNA) has recently been widely studied as the form of RNA useful for therapeutic applications due to its high stability and lowered immune response. Herein, we extend the scope of the recently established RNA-TAG (transglycosylation at guanosine) methodology, a novel approach for genetically encoded site-specific labeling of large mRNA transcripts, by employing mod-mRNA as substrate. As a proof of concept, we covalently attached a fluorescent probe to mCherry encoding mod-mRNA transcripts bearing 5-methylcytidine and/or pseudouridine substitutions with high labeling efficiencies. To provide a versatile labeling methodology with a wide range of possible applications, we employed a two-step strategy for functionalization of the mod-mRNA to highlight the therapeutic potential of this new methodology. We envision that this novel and facile labeling methodology of mod-RNA will have great potential in decorating both coding and noncoding therapeutic RNAs with a variety of diagnostic and functional moieties.

  14. Methods for the synthesis of tritium labelled steroids

    International Nuclear Information System (INIS)

    Volkova, V.S.; Tatarkina, F.V.; Kaklyushkina, L.N.; Ignat'eva, N.A.; Tupitsyn, I.F.; Efimova, Ts.I.

    1977-01-01

    The catalytic substitution of bromine for tritium in corresponding bromo-derivatives of steroids was used for the preparation of 4 steroids labelled with tritium at position 7. The bromination of the starting steroids was effected with N-bromosuccinimide. Ten steroids labelled with tritium at the positions 1, 2, 6 and 7 were prepared by reduction of the unsaturated derivatives with gaseous tritium in the presence of either the heterogeneous Pd/C catalyst, or the homogeneous chloride of tris(triphenylphosphine)rhodium

  15. Optimization under Uncertainty of Site-Specific Turbine Configurations: Preprint

    Energy Technology Data Exchange (ETDEWEB)

    Quick, Julian; Dykes, Katherine; Graf, Peter; Zahle, Frederik

    2016-11-01

    Uncertainty affects many aspects of wind energy plant performance and cost. In this study, we explore opportunities for site-specific turbine configuration optimization that accounts for uncertainty in the wind resource. As a demonstration, a simple empirical model for wind plant cost of energy is used in an optimization under uncertainty to examine how different risk appetites affect the optimal selection of a turbine configuration for sites of different wind resource profiles. If there is unusually high uncertainty in the site wind resource, the optimal turbine configuration diverges from the deterministic case and a generally more conservative design is obtained with increasing risk aversion on the part of the designer.

  16. Optimization Under Uncertainty of Site-Specific Turbine Configurations

    Science.gov (United States)

    Quick, J.; Dykes, K.; Graf, P.; Zahle, F.

    2016-09-01

    Uncertainty affects many aspects of wind energy plant performance and cost. In this study, we explore opportunities for site-specific turbine configuration optimization that accounts for uncertainty in the wind resource. As a demonstration, a simple empirical model for wind plant cost of energy is used in an optimization under uncertainty to examine how different risk appetites affect the optimal selection of a turbine configuration for sites of different wind resource profiles. If there is unusually high uncertainty in the site wind resource, the optimal turbine configuration diverges from the deterministic case and a generally more conservative design is obtained with increasing risk aversion on the part of the designer.

  17. Site-Specific ecological risk assessment. Case-study 2

    DEFF Research Database (Denmark)

    Jensen, John

    “Development of a decision support system for sustainable management of contaminated land by linking bioavailability, ecological risk and ground water pollution of organic pollutants”or in short “LIBERATION”. The presentation includes examples on how to scale and integrate the results from various scientific......The decision supporting and integrating assessment tool, TRIAD, is used site-specific on PAH- and heavy metal contaminated sites in Denmark. The various aspects of the TRIAD approach are used on a set of chemistry-, ecotoxicology- and ecology related data collected among others in the EU project...

  18. Appreciating Site-Specific Qualities in Urban Harbours

    DEFF Research Database (Denmark)

    Reeh, Henrik

    2015-01-01

    of observa-tions from Marseille in southern France. After modernization and dislocation of its harbor territories in the early 20th century already, this city is currently taking important steps from industrial urbanism into cultural planning. This transformation allows for new and unprogrammed experiences......When “site-specificity” becomes a central value in city and harbor transformation, it soon proves necessary to address the ways in which scholars and professionals actually determine site-specific qualities in urban fabrics and social life. This paper delves into the above questions by means...

  19. Optimization of in vitro cell labeling methods for human umbilical cord-derived mesenchymal stem cells.

    Science.gov (United States)

    Tao, R; Sun, T-J; Han, Y-Q; Xu, G; Liu, J; Han, Y-F

    2014-01-01

    Human umbilical cord-derived mesenchymal stem cells (hUCMSCs) are a novel source of seed cells for cell therapy and tissue engineering. However, in vitro labeling methods for hUCMSCs need to be optimized for better detection of transplanted cells. To identify the most stable and efficient method for labeling hUCMSCs in vitro. hUCMSCs were isolated using a modified enzymatic digestion procedure and cultured. hUCMSCs of passage three (P3) were then labeled with BrdU, PKH26, or lentivirus-GFP and passaged further. Cells from the first labeled passage (LP1), the fourth labeled passage (LP4) and later passages were observed using a fluorescence microscope. The differentiation potential of LP4 cells was assessed by induction with adipogenic and osteogenic medium. Flow cytometry was used to measure the percentage of labeled cells and the percentage of apoptotic or dead cells. The labeling efficiencies of the three hUCMSC-labeling methods were compared in vitro. BrdU, PKH26, and lentivirus-GFP all labeled LP1 cells with high intensity and clarity. However, the BrdU labeling of the LP4 cells was vague and not localized to the cell nuclei; LP9 cells were not detected under a fluorescence microscope. There was also a significant decrease in the fluorescence intensity of PKH26-labeled LP4 cells, and LP11 cells were not detected under a fluorescence microscope. However, the fluorescence of LP4 cells labeled with lentivirus-GFP remained strong, and cells labeled with lentivirus-GFP were detected up to LP14 under a fluorescence microscope. Statistical analyses indicated that percentages of LP1 cells labeled with PKH26 and lentivirus-GFP were significantly higher than that of cells labeled with BrdU (p 0.05) was observed between the death rates of labeled and unlabeled cells. Lentivirus-GFP is a valid method for long-term in vitro labeling, and it may be used as a long-term hUCMSC tracker following transplantation in vivo.

  20. Detection of site specific glycosylation in proteins using flow cytometry†

    Science.gov (United States)

    Jayakumar, Deepak; Marathe, Dhananjay D.; Neelamegham, Sriram

    2009-01-01

    We tested the possibility that it is possible to express unique peptide probes on cell surfaces and detect site-specific glycosylation on these peptides using flow cytometry. Such development can enhance the application of flow cytometry to detect and quantify post-translational modifications in proteins. To this end, the N-terminal section of the human leukocyte glycoprotein PSGL-1 (P-selectin glycoprotein ligand-1) was modified to contain a poly-histidine tag followed by a proteolytic cleavage site. Amino acids preceding the cleavage site have a single O-linked glycosylation site. The recombinant protein called PSGL-1 (HT) was expressed on the surface of two mammalian cell lines, CHO and HL-60, using a lentiviral delivery approach. Results demonstrate that the N-terminal portion of PSGL-1 (HT) can be released from these cells by protease, and the resulting peptide can be readily captured and detected using cytometry-bead assays. Using this strategy, the peptide was immunoprecipitated onto beads bearing mAbs against either the poly-histidine sequence or the human PSGL-1. The carbohydrate epitope associated with the released peptide was detected using HECA-452 and CSLEX-1, monoclonal antibodies that recognize the sialyl Lewis-X epitope. Finally, the peptide released from cells could be separated and enriched using nickel chelate beads. Overall, such an approach that combines recombinant protein expression with flow cytometry, may be useful to quantify changes in site-specific glycosylation for basic science and clinical applications. PMID:19735085

  1. Site-specific calibration of the Hanford personnel neutron dosimeter

    International Nuclear Information System (INIS)

    Endres, A.W.; Brackenbush, L.W.; Baumgartner, W.V.; Rathbone, B.A.

    1994-10-01

    A new personnel dosimetry system, employing a standard Hanford thermoluminescent dosimeter (TLD) and a combination dosimeter with both CR-39 nuclear track and TLD-albedo elements, is being implemented at Hanford. Measurements were made in workplace environments in order to verify the accuracy of the system and establish site-specific factors to account for the differences in dosimeter response between the workplace and calibration laboratory. Neutron measurements were performed using sources at Hanford's Plutonium Finishing Plant under high-scatter conditions to calibrate the new neutron dosimeter design to site-specific neutron spectra. The dosimeter was also calibrated using bare and moderated 252 Cf sources under low-scatter conditions available in the Hanford Calibration Laboratory. Dose equivalent rates in the workplace were calculated from spectrometer measurements using tissue equivalent proportional counter (TEPC) and multisphere spectrometers. The accuracy of the spectrometers was verified by measurements on neutron sources with calibrations directly traceable to the National Institute of Standards and Technology (NIST)

  2. Towards soft robotic devices for site-specific drug delivery.

    Science.gov (United States)

    Alici, Gursel

    2015-01-01

    Considerable research efforts have recently been dedicated to the establishment of various drug delivery systems (DDS) that are mechanical/physical, chemical and biological/molecular DDS. In this paper, we report on the recent advances in site-specific drug delivery (site-specific, controlled, targeted or smart drug delivery are terms used interchangeably in the literature, to mean to transport a drug or a therapeutic agent to a desired location within the body and release it as desired with negligibly small toxicity and side effect compared to classical drug administration means such as peroral, parenteral, transmucosal, topical and inhalation) based on mechanical/physical systems consisting of implantable and robotic drug delivery systems. While we specifically focus on the robotic or autonomous DDS, which can be reprogrammable and provide multiple doses of a drug at a required time and rate, we briefly cover the implanted DDS, which are well-developed relative to the robotic DDS, to highlight the design and performance requirements, and investigate issues associated with the robotic DDS. Critical research issues associated with both DDSs are presented to describe the research challenges ahead of us in order to establish soft robotic devices for clinical and biomedical applications.

  3. Methods for labeling .beta.-amyloid plaques and neurofibrillary tangles

    Science.gov (United States)

    Barrio, Jorge R.; Petric, Andrej; Satyamurthy, Nagichettiar; Small, Gary W.; Cole, Gregory M.; Huang, Sung-Cheng

    2001-01-01

    A method for labeling .beta.-amyloid plaques and neurofibrillary tangles in vivo and in vitro, comprises contacting a compound of formula (I): ##STR1## with mammalian tissue. In formula (I), R.sub.1 is selected from the group consisting of --C(O)-alkyl, --C(O)-alkylenyl-R.sub.4, --C(O)O-alkyl, --C(O)O-alkylenyl-R.sub.4, --C.dbd.C(CN).sub.2 -alkyl, --C.dbd.C(CN).sub.2 -alkylenyl-R.sub.4 , ##STR2## R.sub.4 is a radical selected from the group consisting of alkyl, substituted alkyl, aryl and substituted aryl; R.sub.5, is a radical selected from the group consisting of --NH.sub.2, --OH, --SH, --NH-alkyl, --NHR.sub.4, --NH-alkylenyl-R.sub.4, --O-alkyl, --O-alkylenyl-R.sub.4, --S-alkyl, and --S-alkylenyl-R.sub.4 ; R.sub.6 is a radical selected from the group consisting of --CN, --COOH, --C(O)O-alkyl, --C(O)O-alkylenyl-R.sub.4, --C(O)-alkyl, --C(O)-alkylenyl-R.sub.4, --C(O)-halogen, --C(O)NH , --C(O)NH-alkyl, --C(O)NH-alkylenyl-R.sub.4 ; R.sub.7 is a radical selected from the group consisting of O, NH, and S; and R.sub.8 is N, O or S. R.sub.2 and R.sub.3 are each independently selected from the group consisting of alkyl and alkylenyl-R.sub.10, wherein R.sub.10 is selected from the group consisting of --OH, --OTs, halogen, spiperone, spiperone ketal and spiperone-3-yl. Alternatively, R.sub.2 and R.sub.3 together form a heterocyclic ring, optionally substituted with at least one radical selected from the group consisting of alkyl, alkoxy, OH, OTs, halogen, alkylenyl-R.sub.10, carbonyl, spiperone, spiperone ketal and spiperone-3-yl. In the compounds of formula (I), one or more of the hydrogen, halogen or carbon atoms can, optionally, be replaced with a radiolabel.

  4. Site Specific Waste Management Instructions for loading and shipment of category 3 investigation derived waste to ERDF

    International Nuclear Information System (INIS)

    Corriveau, C.E.; Wolf, D.M.

    1996-08-01

    This Site Specific Waste Management Instruction (SSWMI) provides guidance for management of containerized investigation-derived waste being loaded and transported to the Environmental Restoration Disposal Facility. The SSWMI outlines the waste management practices that will be performed in the field to implement federal, state, and US Department of Energy requirements. Additional guidance for waste packaging, marking, labeling and shipping is provided (US DOT rules in 49 CFR have precedence)

  5. A revised method of labeling mouse IgG with yttrium-90

    International Nuclear Information System (INIS)

    Arbab, A.S.; Koizumi, Kiyoshi; Araki, Tsutomu

    1996-01-01

    We report the successful labeling of mouse IgG with yttrium-90 (Y-90) using isothiocyanatobenzyl-ethylenediaminetetraacetic acid (SCN-Bz-EDTA) as a chelating agent and compared the result with labeling by indium-111 (In-111). After conjugating IgG with SCN-Bz-EDTA, a predetermined volume of conjugated IgG was mixed with different volumes of either Y-90 or In-111 acetate and incubated at 37degC. Labeling efficiency was assessed at specific intervals upto 3 hr. After 3 hr, the mixtures were challenged with Na 2 EDTA to evaluate the transchelation of labeled Y-90 or In-111. All mixtures showed labeling efficiency of around 50% with Y-90 and the leveling was fairly preserved even after Na 2 EDTA challenge. However, labeling with In-111 was unsuccessful when conjugated IgG was not separated from the unconjugated form. When separated, however, In-111 showed more than 80% labeling efficiency though labeling with In-111 could not tolerate Na 2 EDTA challenge. In conclusion IgG was efficiently labeled by Y-90 using SCN-Bz-EDTA though labeling with In-111 showed some problems associated with this method. (author)

  6. A simple method for stem cell labeling with fluorine 18

    International Nuclear Information System (INIS)

    Ma Bing; Hankenson, Kurt D.; Dennis, James E.; Caplan, Arnold I.; Goldstein, Steven A.; Kilbourn, Michael R.

    2005-01-01

    Hexadecyl-4-[ 18 F]fluorobenzoate ([ 18 F]HFB), a long chain fluorinated benzoic acid ester, was prepared in a one-step synthesis by aromatic nucleophilic substitution of [ 18 F]fluoride ion on hexadecyl-4-(N,N,N-trimethylammonio)benzoate. The radiolabeled ester was obtained in good yields (52% decay corrected) and high purity (97%). [ 18 F]HFB was used to radiolabel rat mesenchymal stem cells (MSCs) by absorption into cell membranes. MicroPET imaging of [ 18 F]HFB-labeled MSCs following intravenous injection into the rat showed the expected high and persistent accumulation of radioactivity in the lungs. [ 18 F]HFB is thus simple to prepare and uses labeling agent for short-term distribution studies of injected stem cells

  7. Intrinsic mineral labeling of edible plants: methods and uses

    International Nuclear Information System (INIS)

    Weaver, C.M.

    1985-01-01

    The fate of minerals can be conveniently studied through intrinsic labeling techniques. The mineral of interest is biologically incorporated into the food in a form that can be distinguished analytically from the natural form of the element. Radiolabels have traditionally been used to study such problems as the uptake of minerals by plants, the gross and subcellular mineral distribution in plant tissues, the form and associations of the deposited mineral, and the bioavailability of minerals to animals and humans. The use of stable (nonradioactive) isotopes as a label offers the potential of safely studying bioavailability of minerals from individual foods in human population groups of all ages using foods processed in normal food handling and processing facilities. 114 references

  8. A simple method for stem cell labeling with fluorine 18

    Energy Technology Data Exchange (ETDEWEB)

    Ma Bing [Department of Radiology, Division of Nuclear Medicine, University of Michigan Medical School, Ann Arbor, MI 48109 (United States); Hankenson, Kurt D. [Department of Biology, Case Western Reserve University, Cleveland, OH 44106 (United States); Dennis, James E. [Department of Biology, Case Western Reserve University, Cleveland, OH 44106 (United States); Caplan, Arnold I. [Department of Biology, Case Western Reserve University, Cleveland, OH 44106 (United States); Goldstein, Steven A. [Department of Orthopaedic Surgery, University of Michigan Medical School, Ann Arbor, MI 48109 (United States); Kilbourn, Michael R. [Department of Radiology, Division of Nuclear Medicine, University of Michigan Medical School, Ann Arbor, MI 48109 (United States)

    2005-10-01

    Hexadecyl-4-[{sup 18}F]fluorobenzoate ([{sup 18}F]HFB), a long chain fluorinated benzoic acid ester, was prepared in a one-step synthesis by aromatic nucleophilic substitution of [{sup 18}F]fluoride ion on hexadecyl-4-(N,N,N-trimethylammonio)benzoate. The radiolabeled ester was obtained in good yields (52% decay corrected) and high purity (97%). [{sup 18}F]HFB was used to radiolabel rat mesenchymal stem cells (MSCs) by absorption into cell membranes. MicroPET imaging of [{sup 18}F]HFB-labeled MSCs following intravenous injection into the rat showed the expected high and persistent accumulation of radioactivity in the lungs. [{sup 18}F]HFB is thus simple to prepare and uses labeling agent for short-term distribution studies of injected stem cells.

  9. Site specific health and safety plan for drilling in support of in situ redox manipulation

    International Nuclear Information System (INIS)

    Tuttle, B.G.

    1997-02-01

    This document contains the Site Specific Health and Safety Plan for Drilling in support of the In Situ REDOX Manipulation in the 100-HR-3 Operable Unit. Approximately eight wells will be drilled in the 100-D/DR Area using rotary, sonic, or cable tool drilling methods. Split-spoon sampling will be done in conjunction with the drilling. The drilling may be spread out over several months. Included in this document are checklists for health and safety procedures

  10. Bioorthogonal fluorescent labeling of functional G-protein-coupled receptors

    DEFF Research Database (Denmark)

    Tian, He; Naganathan, Saranga; Kazmi, Manija A

    2014-01-01

    Novel methods are required for site-specific, quantitative fluorescent labeling of G-protein-coupled receptors (GPCRs) and other difficult-to-express membrane proteins. Ideally, fluorescent probes should perturb the native structure and function as little as possible. We evaluated bioorthogonal...

  11. Characterization of reference and site specific humic acids

    International Nuclear Information System (INIS)

    Kim, J.I.; Buckau, G.

    1988-11-01

    As a contribution to the interlaboratory exercise for the complexation of humic acid and colloid generation (COCO-Club activities) in the CEC project MIRAGE-II, the characterization of selected humic acids have been carried out at TU Muenchen, regarding their elemental compositions, inorganic impurities, spectroscopic properties, size distributions and proton exchange capacities. The commercial humic acid (Na salt) from Aldrich Co. is purified to a protonated form and used as reference material. Furthermore two humic acids extracted from groundwaters from Gorleben (FRG) and Boom Clay (B) are purified to protonated forms and taken as site specific materials. These three humic acids, together with the original Na salt from Aldrich Co., are included in the present characterization exercise. The results of characterization provide basic knowledge supporting the forthcoming study of complexation of actinides and fission products with humic acid and their migration processes in the geosphere. (orig.)

  12. Site-Specific Genome Engineering in Human Pluripotent Stem Cells.

    Science.gov (United States)

    Merkert, Sylvia; Martin, Ulrich

    2016-06-24

    The possibility to generate patient-specific induced pluripotent stem cells (iPSCs) offers an unprecedented potential of applications in clinical therapy and medical research. Human iPSCs and their differentiated derivatives are tools for diseases modelling, drug discovery, safety pharmacology, and toxicology. Moreover, they allow for the engineering of bioartificial tissue and are promising candidates for cellular therapies. For many of these applications, the ability to genetically modify pluripotent stem cells (PSCs) is indispensable, but efficient site-specific and safe technologies for genetic engineering of PSCs were developed only recently. By now, customized engineered nucleases provide excellent tools for targeted genome editing, opening new perspectives for biomedical research and cellular therapies.

  13. Savannah River Site's Site Specific Plan

    Energy Technology Data Exchange (ETDEWEB)

    1991-08-01

    This Site Specific Plan (SSP) has been prepared by the Savannah River Site (SRS) in order to show the Environmental Restoration and Waste Management activities that were identified during the preparation of the Department of Energy-Headquarters (DOE-HQ) Environmental Restoration and Waste Management Five-Year Plan (FYP) for FY 1992--1996. The SSP has been prepared in accordance with guidance received from DOE-HQ. DOE-SR is accountable to DOE-HQ for the implementation of this plan. The purpose of the SSP is to develop a baseline for policy, budget, and schedules for the DOE Environmental Restoration and Waste Management activities. The plan explains accomplishments since the Fiscal Year (FY) 1990 plan, demonstrates how present and future activities are prioritized, identifies currently funded activities and activities that are planned to be funded in the upcoming fiscal year, and describes future activities that SRS is considering.

  14. Drainage filter technologies to mitigate site-specific phosphorus losses

    DEFF Research Database (Denmark)

    Kjærgaard, Charlotte; Heckrath, Goswin Johann; Iversen, Bo Vangsø

    2014-01-01

    -specific nutrient losses in drainage. The “SUPREME-TECH” project (2010-2015), funded by the Danish Strategic Research Council, aims at providing the scientific basis for developing cost-effective drainage filter technologies to retain P in agricultural drainage waters. The project studies different approaches...... high risks areas of P loss and applying site-specific measures therefore seems a more cost-efficient approach. The Danish Commission for Nature and Agriculture has now called for a shift of paradigm towards targeted mitigation and development of new, cost-efficient technologies to mitigate site......-scale surface-flow constructed wetland. In the former, various natural and industrial P filter substrates have been tested for their ability to reduce inlet P concentrations to below environmental threshold values (

  15. Characterization of reference and site specific human acids

    International Nuclear Information System (INIS)

    Kim, J.I.; Buckau, G.

    1988-01-01

    As a part of the interlaboratory exercise for the complexation of humic acid and colloid generation (COCO-Club activities) in the CEC project MIRAGE-II, the characterization of humic acids have been carried out, as for their elemental compositions, inorganic impurities, spectroscopic properties, size distributions and proton exchange capacities. The commercial humic acid (Na salt) from Aldrich Co. is purified to a protonated form and used as a reference material, and the humic acid extracted from one of Gorleben groundwaters is also purified to a protonated form and taken as a site specific material. These two humic acids, together with the original Na salt from Aldrich Co., are included for the characterization exercise. The results of characterization provide a basic knowledge that supports the forthcoming study of complexation of humic acids with actinides and fission products in their migration processes in the geosphere. (orig.)

  16. Site-specific DNA Inversion by Serine Recombinases

    Science.gov (United States)

    2015-01-01

    Reversible site-specific DNA inversion reactions are widely distributed in bacteria and their viruses. They control a range of biological reactions that most often involve alterations of molecules on the surface of cells or phage. These programmed DNA rearrangements usually occur at a low frequency, thereby preadapting a small subset of the population to a change in environmental conditions, or in the case of phages, an expanded host range. A dedicated recombinase, sometimes with the aid of additional regulatory or DNA architectural proteins, catalyzes the inversion of DNA. RecA or other components of the general recombination-repair machinery are not involved. This chapter discusses site-specific DNA inversion reactions mediated by the serine recombinase family of enzymes and focuses on the extensively studied serine DNA invertases that are stringently controlled by the Fis-bound enhancer regulatory system. The first section summarizes biological features and general properties of inversion reactions by the Fis/enhancer-dependent serine invertases and the recently described serine DNA invertases in Bacteroides. Mechanistic studies of reactions catalyzed by the Hin and Gin invertases are then discussed in more depth, particularly with regards to recent advances in our understanding of the function of the Fis/enhancer regulatory system, the assembly of the active recombination complex (invertasome) containing the Fis/enhancer, and the process of DNA strand exchange by rotation of synapsed subunit pairs within the invertasome. The role of DNA topological forces that function in concert with the Fis/enhancer controlling element in specifying the overwhelming bias for DNA inversion over deletion and intermolecular recombination is emphasized. PMID:25844275

  17. Site Specific Analyses of a Spent Nuclear Fuel Transportation Accident

    International Nuclear Information System (INIS)

    Biwer, B. M.; Chen, S. Y.

    2003-01-01

    The number of spent nuclear fuel (SNF) shipments is expected to increase significantly during the time period that the United States' inventory of SNF is sent to a final disposal site. Prior work estimated that the highest accident risks of a SNF shipping campaign to the proposed geologic repository at Yucca Mountain were in the corridor states, such as Illinois. The largest potential human health impacts would be expected to occur in areas with high population densities such as urban settings. Thus, our current study examined the human health impacts from the most plausible severe SNF transportation accidents in the Chicago metropolitan area. The RISKIND 2.0 program was used to model site-specific data for an area where the largest impacts might occur. The results have shown that the radiological human health consequences of a severe SNF rail transportation accident on average might be similar to one year of exposure to natural background radiation for those persons living a nd working in the most affected areas downwind of the actual accident location. For maximally exposed individuals, an exposure similar to about two years of exposure to natural background radiation was estimated. In addition to the accident probabilities being very low (approximately 1 chance in 10,000 or less during the entire shipping campaign), the actual human health impacts are expected to be lower if any of the accidents considered did occur, because the results are dependent on the specific location and weather conditions, such as wind speed and direction, that were selected to maximize the results. Also, comparison of the results of longer duration accident scenarios against U.S. Environmental Protection Agency guidelines was made to demonstrate the usefulness of this site-specific analysis for emergency planning purposes

  18. SORIOS – A method for evaluating and selecting environmental certificates and labels

    DEFF Research Database (Denmark)

    Kikkenborg Pedersen, Dennis; Dukovska-Popovska, Iskra; Ola Strandhagen, Jan

    2012-01-01

    This paper presents a general method for evaluating and selecting environmental certificates and labels for companies to use on products and services. The method is developed based on a case study using a Grounded Theory approach. The result is a generalized six-step method that features an initial...... searching strategy and an evaluation model that weighs the prerequisites, rewards and the organization of certificate or label against the strategic needs of a company....

  19. Technetium 99-m labeled radio-diagnostic agents employing stannous tartrate and method of preparation

    International Nuclear Information System (INIS)

    Molinski, V.J.; Wilczewski, J.A.

    1976-01-01

    A method of preparing improved technetium-99m labeled radiodiagnostic agents by reducing technetium-99m with stannous tartrate. Such radiodiagnostic agents are useful in scintigraphic examinations of the bone and lung. 31 claims, no drawings

  20. A comparative study on the iodine-labeled methods of protein and polypeptide

    International Nuclear Information System (INIS)

    Li Huaifen; Niu Huisheng; Yuan Mingyue; Yu Jinghua

    1994-01-01

    There are three methods: chloramine-T, Iodogen and lactoperoxidase(LPO). 125 I-ACTH, 125 I-insulin and 125 I-HSA are prepared by these techniques. The results show that lactoperoxidase is isolated and purified from fresh milk, meanwhile, the enzyme is used in experiments of 125 I-labeled protein, peptide hormone and mono-clone antibody, etc. LPO is a very successful method for it's mild, complete reaction, controllable, high labelling yield, higher purity of iodine-labeled compound and so on. It remains biological activation and stable character more than other two techniques

  1. A comparative study on the iodine-labeled methods of protein and polypeptide

    Energy Technology Data Exchange (ETDEWEB)

    Huaifen, Li; Huisheng, Niu; Mingyue, Yuan; Jinghua, Yu [Chinese Academy of Medical Sciences, Tianjin (China). Inst. of Radiation Medicine

    1994-02-01

    There are three methods: chloramine-T, Iodogen and lactoperoxidase(LPO). [sup 125]I-ACTH, [sup 125]I-insulin and [sup 125]I-HSA are prepared by these techniques. The results show that lactoperoxidase is isolated and purified from fresh milk, meanwhile, the enzyme is used in experiments of [sup 125]I-labeled protein, peptide hormone and mono-clone antibody, etc. LPO is a very successful method for it's mild, complete reaction, controllable, high labelling yield, higher purity of iodine-labeled compound and so on. It remains biological activation and stable character more than other two techniques.

  2. Site-specific estimates of water yield applied in regional acid sensitivity surveys across western Canada

    Directory of Open Access Journals (Sweden)

    Patrick D. SHAW

    2010-08-01

    Full Text Available Runoff or water yield is an important input to the Steady-State Water Chemistry (SSWC model for estimating critical loads of acidity. Herein, we present site-specific water yield estimates for a large number of lakes (779 across three provinces of western Canada (Manitoba, Saskatchewan, and British Columbia using an isotope mass balance (IMB approach. We explore the impact of applying site-specific hydrology as compared to use of regional runoff estimates derived from gridded datasets in assessing critical loads of acidity to these lakes. In general, the average water yield derived from IMB is similar to the long-term average runoff; however, IMB results suggest a much larger range in hydrological settings of the lakes, attributed to spatial heterogeneity in watershed characteristics and landcover. The comparison of critical loads estimates from the two methods suggests that use of average regional runoff data in the SSWC model may overestimate critical loads for the majority of lakes due to systematic skewness in the actual runoff distributions. Implications for use of site-specific hydrology in regional critical loads assessments across western Canada are discussed.

  3. Site-specific cancer risk in the Baltic cohort of Chernobyl cleanup workers, 1986–2007

    Science.gov (United States)

    Rahu, Kaja; Hakulinen, Timo; Smailyte, Giedre; Stengrevics, Aivars; Auvinen, Anssi; Inskip, Peter D.; Boice, John D.; Rahu, Mati

    2013-01-01

    Objective To assess site-specific cancer risk in the Baltic cohort of Chernobyl cleanup workers 1986–2007. Methods The Baltic cohort includes 17,040 men from Estonia, Latvia and Lithuania who participated in the environmental cleanup after the accident at the Chernobyl Nuclear Power Station in 1986–1991, and who were followed for cancer incidence until the end of 2007. Cancer cases diagnosed in the cohort and in the male population of each country were identified from the respective national cancer registers. The proportional incidence ratio (PIR) with 95% confidence interval (CI) was used to estimate the site-specific cancer risk in the cohort. For comparison and as it was possible, the site-specific standardized incidence ratio (SIR) was calculated for the Estonian sub-cohort, which was not feasible for the other countries. Results Overall, 756 cancer cases were reported during 1986–2007. A higher proportion of thyroid cancers in relation to the male population was found (PIR=2.76; 95%CI 1.63–4.36), especially among those who started their mission shortly after the accident, in April–May 1986 (PIR=6.38; 95% CI 2.34–13.89). Also, an excess of oesophageal cancers was noted (PIR=1.52; 95% CI 1.06–2.11). No increased PIRs for leukaemia or radiation-related cancer sites combined were observed. PIRs and SIRs for the Estonian sub-cohort demonstrated the same site-specific cancer risk pattern. Conclusion Consistent evidence of an increase in radiation-related cancers in the Baltic cohort was not observed with the possible exception of thyroid cancer, where conclusions are hampered by known medical examination including thyroid screening among cleanup workers. PMID:23683549

  4. DAFS study of site-specific local structure of Mn in manganese ferrite films

    International Nuclear Information System (INIS)

    Kravtsov, E.; Haskel, D.; Cady, A.; Yang, A.; Vittoria, C.; Zuo, X.; Harris, V.G.

    2006-01-01

    Manganese ferrite (MnFe 2 O 4 ) is a well-known magnetic material widely used in electronics for many years. It is well established that its magnetic behavior is strongly influenced by local structural properties of Mn ions, which are distributed between crystallographically inequivalent tetrahedral and octahedral sites in the unit cell. In order to understand and be able to tune properties of these structures, it is necessary to have detailed site-specific structural information on the system. Here we report on the application of diffraction-anomalous fine structure (DAFS) spectroscopy to resolve site-specific Mn local structures in manganese ferrite films. The DAFS measurements were done at undulator beamline 4-ID-D of the Advanced Photon Source at Argonne National Laboratory. The DAFS spectra (Fig. 1) were measured at several Bragg reflections in the vicinity of the Mn absorption K-edge, having probed separately contributions from tetrahedrally and octahedrally coordinated Mn sites. The DAFS data analysis done with an iterative Kramers-Kroenig algorithm made it possible to solve separately the local structure around different inequivalent Mn sites in the unit cell. The reliability of the data treatment was checked carefully, and it was showed that the site-specific structural parameters obtained with DAFS allow us to describe fluorescence EXAFS spectrum measured independently. Fig. 2 shows individual site contributions to the imaginary part of the resonant scattering amplitude obtained from the treatment of the data of Fig. 1. The analysis of the refined site-specific absorption spectra was done using EXAFS methods based on theoretical standards. We provided direct evidence for the tetrahedral Mn-O bond distance being increased relative to the corresponding Fe-O distance in bulk manganese ferrites. The first coordination shell number was found to be reduced significantly for Mn atoms at these sites. This finding is consistent with the well-known tendency of Mn

  5. The method of obtaining of sodium orthoiodohippurate labelled with iodine-131

    International Nuclear Information System (INIS)

    Aripov, D.; Abdukayumov, M.; Shukurov, A.Sh.

    1994-01-01

    The method of labelling of sodium orthoiodohippurate was elaborated with the purpose of increasing the preparation quality. Method includes the reaction of isotopic exchange between orthoiodhippur acid and sodium iodide solution labelled with iodine-131 with volume activity 150-200 mCu/mL and pH=6,5-7,0. Reaction occurs at temperature 120-130 C during 1,1-1,3 hours and the compound obtained is dissolved in 1% sodium bicarbonate solution. (author)

  6. Phosphine reduced IgG. A new method for 99mTc labeling immunoglobulins

    International Nuclear Information System (INIS)

    Arteaga de Murphy, C.; Melendez-Alafort, L.; Martinez-Rivero, O.; Gomez, E.; Ferro-Flores, G.

    1997-01-01

    A new technetium labeling method for immunoglobulins reduced with tris(2-carboxy-ethyl)phosphine hydrochloride is presented. The Sandoglobulina IgG source was assayed for purity and optimum reagent's concentration and incubation times were determined. It was purified by column chromatography and labelled with Sn 2+ reduced technetium in the presence of MDP. The kit is easy to prepare, labeling efficiency is >(97±1.9)% and stable for 6 hours.The immunoreactivity of the 99 Tc-IgG was verified by electrophoresis and Western blot tests. The IgG retained its structure after both the reducing and labeling processes and it was the only labeled species. (author)

  7. Phosphine reduced IgG. A new method for {sup 99m}Tc labeling immunoglobulins

    Energy Technology Data Exchange (ETDEWEB)

    Arteaga de Murphy, C; Melendez-Alafort, L [Radiofarmacia Departamento de Medicina Nuclear, Instituto Nacional de Nutricion Salvador Zubiran, Mexico (Mexico); Martinez-Rivero, O [Laboratorio de Quimica Organica, Facultad de Quimica, Universidad de la Habana, Habana (Cuba); Gomez, E [Departamento de Fisiologia de la Nutricion, Instituto Nacional de Nutricion Salvador Zubiran, Mexico (Mexico); Ferro-Flores, G [Depeartamento del Reactor y Materiales Radioactivos, Instituto Nacional de Investigaciones Nucleares, Mexico (Mexico)

    1997-09-01

    A new technetium labeling method for immunoglobulins reduced with tris(2-carboxy-ethyl)phosphine hydrochloride is presented. The Sandoglobulina IgG source was assayed for purity and optimum reagent`s concentration and incubation times were determined. It was purified by column chromatography and labelled with Sn{sup 2+} reduced technetium in the presence of MDP. The kit is easy to prepare, labeling efficiency is >(97{+-}1.9)% and stable for 6 hours.The immunoreactivity of the {sup 99}Tc-IgG was verified by electrophoresis and Western blot tests. The IgG retained its structure after both the reducing and labeling processes and it was the only labeled species. (author). 11 refs.

  8. Polymerase synthesis of oligonucleotides containing a single chemically modified nucleobase for site-specific redox labelling

    Czech Academy of Sciences Publication Activity Database

    Ménová, Petra; Cahová, Hana; Plucnara, Medard; Havran, Luděk; Fojta, Miroslav; Hocek, Michal

    2013-01-01

    Roč. 49, č. 41 (2013), s. 4652-4654 ISSN 1359-7345 R&D Projects: GA ČR GBP206/12/G151 Institutional support: RVO:61388963 ; RVO:68081707 Keywords : cross - coupling reactions * DNA-protein interactions * nucleoside triphosphates * enzymatic incorporation Subject RIV: CC - Organic Chemistry Impact factor: 6.718, year: 2013

  9. The Application of TAPM for Site Specific Wind Energy Forecasting

    Directory of Open Access Journals (Sweden)

    Merlinde Kay

    2016-02-01

    Full Text Available The energy industry uses weather forecasts for determining future electricity demand variations due to the impact of weather, e.g., temperature and precipitation. However, as a greater component of electricity generation comes from intermittent renewable sources such as wind and solar, weather forecasting techniques need to now also focus on predicting renewable energy supply, which means adapting our prediction models to these site specific resources. This work assesses the performance of The Air Pollution Model (TAPM, and demonstrates that significant improvements can be made to only wind speed forecasts from a mesoscale Numerical Weather Prediction (NWP model. For this study, a wind farm site situated in North-west Tasmania, Australia was investigated. I present an analysis of the accuracy of hourly NWP and bias corrected wind speed forecasts over 12 months spanning 2005. This extensive time frame allows an in-depth analysis of various wind speed regimes of importance for wind-farm operation, as well as extreme weather risk scenarios. A further correction is made to the basic bias correction to improve the forecast accuracy further, that makes use of real-time wind-turbine data and a smoothing function to correct for timing-related issues. With full correction applied, a reduction in the error in the magnitude of the wind speed by as much as 50% for “hour ahead” forecasts specific to the wind-farm site has been obtained.

  10. Site-specific meteorology identification for DOE facility accident analysis

    International Nuclear Information System (INIS)

    Rabin, S.B.

    1995-01-01

    Currently, chemical dispersion calculations performed for safety analysis of DOE facilities assume a Pasquill D-Stability Class with a 4.5 m/s windspeed. These meteorological conditions are assumed to conservatively address the source term generation mechanism as well as the dispersion mechanism thereby resulting in a net conservative downwind consequence. While choosing this Stability Class / Windspeed combination may result in an overall conservative consequence, the level of conservative can not be quantified. The intent of this paper is to document a methodology which incorporates site-specific meteorology to determine a quantifiable consequence of a chemical release. A five-year meteorological database, appropriate for the facility location, is utilized for these chemical consequence calculations, and is consistent with the approach used for radiological releases. The hourly averages of meteorological conditions have been binned into 21 groups for the chemical consequence calculations. These 21 cases each have a probability of occurrence based on the number of times each case has occurred over the five year sampling period. A code has been developed which automates the running of all the cases with a commercially available air modeling code. The 21 cases are sorted by concentration. A concentration may be selected by the user for a quantified level of conservatism. The methodology presented is intended to improve the technical accuracy and defensability of Chemical Source Term / Dispersion Safety Analysis work. The result improves the quality of safety analyses products without significantly increasing the cost

  11. A new method for the labelling of proteins with radioactive arsenic isotopes

    Energy Technology Data Exchange (ETDEWEB)

    Jennewein, M. [Institute of Nuclear Chemistry, Johannes Gutenberg University of Mainz, Fritz-Strassmann-Weg 2, 55128 Mainz (Germany); Hermanne, A. [VUB Cyclotron, University of Brussels, Laarbeeklaan 103, 1090 Brussels (Belgium); Mason, R.P. [Department of Radiology, Advanced Radiological Sciences, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas (United States); Thorpe, P.E. [Department of Pharmacology and Simmons and Hamon Cancer Centers, University of Texas Southwestern Medical Center at Dallas, Dallas, TX (United States); Roesch, F. [Institute of Nuclear Chemistry, Johannes Gutenberg University of Mainz, Fritz-Strassmann-Weg 2, 55128 Mainz (Germany)]. E-mail: frank.roesch@uni-mainz.de

    2006-12-20

    Radioarsenic labelled radiopharmaceuticals could be a valuable asset to positron emission tomography. In particular, the long half-lives of {sup 72}As (T{sub 1/2}=26h) and {sup 74}As (T{sub 1/2}=17.8d) allow to investigate slow physiological or metabolical processes, like the enrichment and distribution of monoclonal antibodies (mab) in tumour tissue. In this work, a new method for the labelling of proteins with various radioactive arsenic isotopes was developed. For this purpose, two proteins, namely a chimeric IgG{sub 3} monoclonal antibody, ch3G4, directed against anionic phospholipids, and Rituxan (Rituximab), were labelled as a proof of principle with no-carrier-added radioarsenic isotopes ({sup 74}As and {sup 77}As). The developed labelling chemistry gives high yields (>99.9%), is reliable and could easily be transferred to automated labelling systems in a clinical environment. At least for the mab used in this work, this route of radioarsenic labelling does not affect the immunoreactivity of the product. The arsenic label stays stable for up to 72h at the molecular mass of the monoclonal antibody, which is in particular relevant to follow the pharmacology and pharmacokinetics of the labelled mab for several days.

  12. Alcohol Warning Label Awareness and Attention: A Multi-method Study.

    Science.gov (United States)

    Pham, Cuong; Rundle-Thiele, Sharyn; Parkinson, Joy; Li, Shanshi

    2018-01-01

    Evaluation of alcohol warning labels requires careful consideration ensuring that research captures more than awareness given that labels may not be prominent enough to attract attention. This study investigates attention of current in market alcohol warning labels and examines whether attention can be enhanced through theoretically informed design. Attention scores obtained through self-report methods are compared to objective measures (eye-tracking). A multi-method experimental design was used delivering four conditions, namely control, colour, size and colour and size. The first study (n = 559) involved a self-report survey to measure attention. The second study (n = 87) utilized eye-tracking to measure fixation count and duration and time to first fixation. Analysis of Variance (ANOVA) was utilized. Eye-tracking identified that 60% of participants looked at the current in market alcohol warning label while 81% looked at the optimized design (larger and red). In line with observed attention self-reported attention increased for the optimized design. The current study casts doubt on dominant practices (largely self-report), which have been used to evaluate alcohol warning labels. Awareness cannot be used to assess warning label effectiveness in isolation in cases where attention does not occur 100% of the time. Mixed methods permit objective data collection methodologies to be triangulated with surveys to assess warning label effectiveness. Attention should be incorporated as a measure in warning label effectiveness evaluations. Colour and size changes to the existing Australian warning labels aided by theoretically informed design increased attention. © The Author 2017. Medical Council on Alcohol and Oxford University Press. All rights reserved.

  13. Site specific incorporation of heavy atom-containing unnatural amino acids into proteins for structure determination

    Science.gov (United States)

    Xie, Jianming [San Diego, CA; Wang, Lei [San Diego, CA; Wu, Ning [Boston, MA; Schultz, Peter G [La Jolla, CA

    2008-07-15

    Translation systems and other compositions including orthogonal aminoacyl tRNA-synthetases that preferentially charge an orthogonal tRNA with an iodinated or brominated amino acid are provided. Nucleic acids encoding such synthetases are also described, as are methods and kits for producing proteins including heavy atom-containing amino acids, e.g., brominated or iodinated amino acids. Methods of determining the structure of a protein, e.g., a protein into which a heavy atom has been site-specifically incorporated through use of an orthogonal tRNA/aminoacyl tRNA-synthetase pair, are also described.

  14. Localized conformational interrogation of antibody and antibody-drug conjugates by site-specific carboxyl group footprinting.

    Science.gov (United States)

    Pan, Lucy Yan; Salas-Solano, Oscar; Valliere-Douglass, John F

    Establishing and maintaining conformational integrity of monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs) during development and manufacturing is critical for ensuring their clinical efficacy. As presented here, we applied site-specific carboxyl group footprinting (CGF) for localized conformational interrogation of mAbs. The approach relies on covalent labeling that introduces glycine ethyl ester tags onto solvent-accessible side chains of protein carboxylates. Peptide mapping is used to monitor the labeling kinetics of carboxyl residues and the labeling kinetics reflects the conformation or solvent-accessibility of side chains. Our results for two case studies are shown here. The first study was aimed at defining the conformational changes of mAbs induced by deglycosylation. We found that two residues in C H 2 domain (D268 and E297) show significantly enhanced side chain accessibility upon deglycosylation. This site-specific result highlighted the advantage of monitoring the labeling kinetics at the amino acid level as opposed to the peptide level, which would result in averaging out of highly localized conformational differences. The second study was designed to assess conformational effects brought on by conjugation of mAbs with drug-linkers. All 59 monitored carboxyl residues displayed similar solvent-accessibility between the ADC and mAb under native conditions, which suggests the ADC and mAb share similar side chain conformation. The findings are well correlated and complementary with results from other assays. This work illustrated that site-specific CGF is capable of pinpointing local conformational changes in mAbs or ADCs that might arise during development and manufacturing. The methodology can be readily implemented within the industry to provide comprehensive conformational assessment of these molecules.

  15. Improvement of organic compounds labelling method with the use of thermally activated tritium gas

    International Nuclear Information System (INIS)

    Nejman, L.A.; Smolyakov, V.S.; Antropova, L.P.

    1982-01-01

    Use of a support (various types of papers) is recommended for organic compounds labelling by tritium gas activated at a hot tungsten filament. This improvement increases chemical and radiochemical yields and makes the experiment simpler and faster. Generally labelled triethyloxonium tetra-fluoroborate, ethyl-p-aminobenzoate, p-aminobenzoic acid (Na-salt), A-factor (a natural regulator of streptomycin biosynthesis), decapeptide angiotensin I, phospholipid 1, 2 - dimyristoyl-sn-glycero-3--phosphocholine and E. coli tRNAs have been prepared by this method. Molar radioactivity of the labelled compounds is in the range of 1-200 GBg/mmole [ru

  16. Site Specific Nutrient Management for Maize on Ultisols Lampung

    Directory of Open Access Journals (Sweden)

    Andarias Makka Murni

    2010-01-01

    Full Text Available Lampung is the third major maize producing province in Indonesia after East Java and Central Java. In Lampungmaize is cultivated mainly in upland areas with ultisols and only some cultivated on paddy field as a secondary cropin the dry season. The average maize yield in Lampung is still 3.4 Mg ha-1 bellow yield potential of 7 - 10 Mg ha-1. Toincrease the productivity of maize through site-specific nutrient management (SSNM, on-farm trials were conductedin five locations in Lampung i.e. four locations in Central Lampung District (Sidowaras, Binjai Ngagung, Watu Agungand Balai Rejo and one location in South Lampung District (Trimulyo, Tegineneng Sub District during the 2004/2005,2005/2006 and 2006/2007 rainy seasons. The experimental setup followed a standard protocol at all sites and includednutrient omission plots (PK, NK, NP to estimate indigenous nutrient supplies, an NPK plot to measure yield responseto fertilizer application, and a farmers’ fertilizer practice (FFP plot in each farmer’s field. An SSNM treatment plot wasincluded in the second and third seasons. Each of the above treatments was paralleled by a plot with improved cropmanagement practice (ICM, i.e. higher planting density, addition of lime, and addition of magnesium. Results showedthat yield response to fertilizer N, P and K application in these sites were: N = 2.3 - 4.1 Mg ha-1; P = 0.6 - 2.0 Mg ha-1;K = 0.3-2.4 Mg ha-1. Attainable yield in the three seasons on average ranged from 7.6 Mg ha-1 to 10.6 Mg ha-1. Yield inthe SSNM treatment (with or without ICM was significantly higher than the FFP indicating great opportunities forfarmers to increase productivity and profitability with improved nutrient and crop management

  17. A comparative study of fat storage quantitation in nematode Caenorhabditis elegans using label and label-free methods.

    Directory of Open Access Journals (Sweden)

    Kelvin Yen

    to the staining of fat stores, but rather the sequestration of dyes in lysosome-related organelles. In contrast, fixative staining methods provide reproducible data but are prone to errors due to the interference of autofluorescent species and the non-specific staining of cellular structures other than fat stores. Importantly, both growth conditions and developmental stage should be considered when comparing methods of C. elegans lipid storage. Taken together, we confirm that CARS microscopy provides a direct, non-invasive, and label-free means to quantitatively analyze fat storage in living C. elegans.

  18. A SIMPLE FLUORESCENT LABELING METHOD FOR STUDIES OF PROTEIN OXIDATION, PROTEIN MODIFICATION, AND PROTEOLYSIS

    Science.gov (United States)

    Pickering, Andrew. M.; Davies, Kelvin. J. A.

    2014-01-01

    Proteins are sensitive to oxidation, and oxidized proteins are excellent substrates for degradation by proteolytic enzymes such as the Proteasome and the mitochondrial Lon protease. Protein labeling is required for studies of protein turnover. Unfortunately, most labeling techniques involve 3H or 14C methylation which is expensive, exposes researchers to radioactivity, generates large amounts of radioactive waste, and allows only single-point assays because samples require acid-precipitation. Alternative labeling methods, have largely proven unsuitable, either because the probe itself is modified by the oxidant(s) being studied, or because the alternative labeling techniques are too complex or too costly for routine use. What is needed is a simple, quick, and cheap labeling technique that uses a non-radioactive marker, that binds strongly to proteins, is resistant to oxidative modification, and emits a strong signal. We have devised a new reductive method for labeling free carboxyl groups of proteins with the small fluorophore 7-amino-4-methycoumarin (AMC). When bound to target proteins, AMC fluoresces very weakly but when AMC is released by proteinases, proteases, or peptidases, it fluoresces strongly. Thus, without acid-precipitation, the proteolysis of any target protein can be studied continuously, in multiwell plates. In direct comparisons, 3H-labeled proteins and AMC-labeled proteins exhibited essentially identical degradation patterns during incubation with trypsin, cell extracts, and purified proteasome. AMC-labeled proteins are well-suited to study increased proteolytic susceptibility following protein modification, since the AMC-protein bond is resistant to oxidizing agents such as hydrogen peroxide and peroxynitrite, and is stable over time and to extremes of pH, temperature (even boiling), freeze-thawing, mercaptoethanol, and methanol. PMID:21988844

  19. 75 FR 7577 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2010-02-22

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Portsmouth AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act... areas of environmental restoration, waste management and related activities. Tentative Agenda: Call to...

  20. 75 FR 65615 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2010-10-26

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Portsmouth AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act... areas of environmental restoration, waste management and related activities. Tentative Agenda Call to...

  1. 76 FR 57981 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2011-09-19

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Portsmouth AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act... environmental restoration, waste management and related activities. Tentative Agenda Call to Order...

  2. 77 FR 2283 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2012-01-17

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Portsmouth AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act... environmental restoration, waste management and related activities. Tentative Agenda Call to Order...

  3. 76 FR 36100 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2011-06-21

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Portsmouth AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act... environmental restoration, waste management and related activities. Tentative Agenda Call to Order...

  4. 76 FR 17118 - Environmental Management Site-Specific Advisory Board Chairs

    Science.gov (United States)

    2011-03-28

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board Chairs AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB) Chairs. The Federal Advisory Committee Act (Pub... areas of environmental restoration, waste management, and related activities. Tentative Agenda Topics...

  5. 76 FR 62054 - Environmental Management Site-Specific Advisory Board Chairs

    Science.gov (United States)

    2011-10-06

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board Chairs AGENCY... of the Environmental Management Site-Specific Advisory Board (EM SSAB) Chairs. The Federal Advisory... environmental restoration, waste management, and related activities. Tentative Agenda Topics [cir] EM Program...

  6. 77 FR 29997 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2012-05-21

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Portsmouth AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act... environmental restoration, waste management and related activities. Tentative Agenda Call to Order...

  7. 77 FR 37390 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2012-06-21

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Portsmouth AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act... environmental restoration, waste management and related activities. Tentative Agenda: Call to Order...

  8. 75 FR 82003 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2010-12-29

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Portsmouth AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act... areas of environmental restoration, waste management and related activities. Tentative Agenda: Call to...

  9. 75 FR 19379 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2010-04-14

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Portsmouth AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act... areas of environmental restoration, waste management and related activities. Tentative Agenda Call to...

  10. 76 FR 78909 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2011-12-20

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Portsmouth AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act... the areas of environmental restoration, waste management, and related activities. Tentative Agenda...

  11. 76 FR 50204 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2011-08-12

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Nevada AGENCY...-Wide Environmental Impact Statement (EIS) Committee of the Environmental Management Site- Specific... management in the areas of environmental restoration, waste management, and related activities. Purpose of...

  12. 77 FR 6790 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2012-02-09

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Portsmouth AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act... environmental restoration, waste management and related activities. Tentative Agenda Call to Order...

  13. 76 FR 55370 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2011-09-07

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Nevada AGENCY...-Wide Environmental Impact Statement (EIS) Committee of the Environmental Management Site- Specific... the areas of environmental restoration, waste management, and related activities. Purpose of the...

  14. 75 FR 51026 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2010-08-18

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Portsmouth AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act... the areas of environmental restoration, waste management and related activities. Tentative Agenda...

  15. 77 FR 51789 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2012-08-27

    ... management and related activities. Tentative Agenda Call to Order, Introductions, Review of Agenda... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Paducah AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act...

  16. Site-Specific Seismic Site Response Model for the Waste Treatment Plant, Hanford, Washington

    Energy Technology Data Exchange (ETDEWEB)

    Rohay, Alan C.; Reidel, Steve P.

    2005-02-24

    This interim report documents the collection of site-specific geologic and geophysical data characterizing the Waste Treatment Plant site and the modeling of the site-specific structure response to earthquake ground motions.

  17. Extraction method for the determination of inorganic iodides in Rose Bengal labelled with 131I

    International Nuclear Information System (INIS)

    Lengyel, J.; Krtil, J.; Vecernik, J.

    1982-01-01

    An extraction method for the determination of inorganic iodides in Rose Bengal preparations labelled with 131 I is described. The method is based on the quantitative extraction of Rose Bengal into chloroform from acidic medium while the inorganic iodides remain in the aqueous phase. The method is simple, rapid, and reproducible. (author)

  18. Site-specific electronic structure analysis by channeling EELS and first-principles calculations.

    Science.gov (United States)

    Tatsumi, Kazuyoshi; Muto, Shunsuke; Yamamoto, Yu; Ikeno, Hirokazu; Yoshioka, Satoru; Tanaka, Isao

    2006-01-01

    Site-specific electronic structures were investigated by electron energy loss spectroscopy (EELS) under electron channeling conditions. The Al-K and Mn-L(2,3) electron energy loss near-edge structure (ELNES) of, respectively, NiAl2O4 and Mn3O4 were measured. Deconvolution of the raw spectra with the instrumental resolution function restored the blunt and hidden fine features, which allowed us to interpret the experimental spectral features by comparing with theoretical spectra obtained by first-principles calculations. The present method successfully revealed the electronic structures specific to the differently coordinated cationic sites.

  19. A rapid and convenient method for specific 11C-labelling of synthetic polypeptides containing methionine

    International Nuclear Information System (INIS)

    Laengstroem, B.; Sjoeberg, S.; Ragnarsson, U.

    1981-01-01

    11 C-labelling of methionine residues in a synthetic peptide via the preparation of the corresponding protected, pure homocysteine peptide has been investigated. Complete deprotection of the peptide and specific methylation of the homocysteine residue can be performed in one step in liquid ammonia. As a first application of this method the synthesis of the tripeptide, Z-Gly-L-Hcy(Bzl)-Gly-O-Bzl, and its conversion to Gly-Met-Gly and the corresponding labelled Gly-([ 11 C]-methyl)-Met-Gly, is reported. Starting with the protected peptide the labelling was performed in 20 +- 5 min (starting with 11 CO 2 ), yielding the labelled peptide in 92 +- 5 % radiochemical yield. Analyses and preparative LC can be performed within 6 min. (author)

  20. Chemical method of labelling proteins with the radionuclides of technetium at physiological condition

    International Nuclear Information System (INIS)

    Wong, D.W.

    1983-01-01

    A novel rapid chemical method of labeling plasma proteins, other compounds and/or substances containing protein with radionuclides of technetium such as sup(95m)Tc, sup(99m)Tc or sup(99)Tc at physiologic pH 7.4 condition, producing a sterile non-pyrogenic radioactive tracer material suitable for biological and medical uses. These radiolabeled protein substances are not denatured by the labeling process but retain their natural physiological and immunological properties. This novel labeling technique provides a simple and rapid means of labeling plasma proteins such as human serum albumin, fibrinogen, antibodies, hormones and enzymes with sup(95m)Tc or sup(99m)Tc for scintigraphic imaging which may allow visualization of thrombi, emboli, myocardial infarcts, infectious lesions or tumors

  1. A rapid method for the preparation of 99Tcm hexametazime-labelled leucocytes

    International Nuclear Information System (INIS)

    Solanki, K.K.; Mather, S.J.; Janabi, M.A.; Britton, K.E.

    1988-01-01

    99 Tc m (±)-hexamethylpropyleneamineoxime (HMPAO) ( 99 Tc m Hexametazime) has been recently reported as an alternative for labelling leucocytes. This technique has been modified to give a simpler routine in-house labelling technique. It has three advantages: only about 20 ml of blood is required, the labelling time is just under 1 h and high yields of labelled leucocytes are obtained (mean of 500 MBq per injection dose). The properties of labelled leucocytes using this modified method are; 80% granulocyte-bound radioactivity, a rapid lung transit and a blood granulocyte recovery of 40% at 30 min similar to those described previously. The viability of the labelled leucocytes was tested and confirmed in vitro using a migration technique and in vivo by showing no lung retention on early imaging and high splenic uptake. A rapid in-process chromatography assessment procedure for regulating the protocol has been developed. Successful abscess imaging by 4 h has been achieved in 21 patients with normal results in another 22 patients without abscesses. This simpler method should encourage a more widespread application of scintigraphy using radiolabelled granulocytes. (author)

  2. Option-4 algorithm for Florida pocket depth probe: reduction in the variance of site-specific probeable crevice depth measurements.

    Science.gov (United States)

    Breen, H J; Rogers, P; Johnson, N W; Slaney, R

    1999-08-01

    Clinical periodontal measurement is plagued by many sources of error which result in aberrant values (outliers). This study sets out to compare probeable crevice depth measurements (PCD) selected by the option-4 algorithm against those recorded with a conventional double-pass method and to quantify any reduction in site-specific PCD variances. A single clinician recorded full-mouth PCD at 1 visit in 32 subjects (mean age 45.5 years) with moderately advanced chronic adult periodontitis. PCD was recorded over 2 passes at 6 sites per tooth with the Florida Pocket Depth Probes, a 3rd generation probe. The option-4 algorithm compared the 1st pass site-specific PCD value (PCD1) to the 2nd pass site-specific PCD value (PCD2) and, if the difference between these values was >1.00 mm, allowed the recording of a maximum of 2 further measurements (3rd and 4th pass measurements PCD3 and PCD4): 4 site-specific measure-meets were considered to be the maximum subject and tissue tolerance. The algorithm selected the 1st 2 measurements whose difference was difference Y) (Y=[(A-B)/A]X 100) and a 75% reduction in the median site-specific variance of PCD1/PCD2.

  3. A comparison of two methods of labelling autologous platelets with 111In-oxine in five different species

    International Nuclear Information System (INIS)

    Christenson, J.T.; Arvidsson, D.; Thoerne, J.; Norgren, L.; Olsson, P.I.; Strand, S.E.

    1983-01-01

    Several different methods for labelling autologous platelets with 111 In-oxine have been described. However, no comparative study has been reported. In the present investigation two different labelling methods were compared in terms of labelling efficiency and platelet function in five species: human, dog, pig, rabbit and rat. One of the labelling methods, utilising among other things a serum albumin gradient separation fo platelets and incubation of 111 In-oxine in a water bath at 37 0 C, was superior in all species with significantly higher labelling efficiency and unchanged platelet function. (orig.)

  4. Doubly labeled water (3HH18O) method: a guide to its use

    International Nuclear Information System (INIS)

    Nagy, K.A.

    1983-06-01

    The doubly labelled water method for measuring CO 2 production and water flux rates is used in studies of energy and material balance in animals living in their natural habitats. This report presents guidelines for the use of this method, including preparation of materials, field and laboratory procedures, and tritium and 18 O analysis techniques

  5. A Mild Method for Regioselective Labeling of Aromatics with Radioactive Iodine

    DEFF Research Database (Denmark)

    Rønnest, Mads Holger; Nissen, Felix; Pedersen, Palle Jacob

    2013-01-01

    A novel technique to label ortho‐, meta‐, and para‐trimethylsilyl‐substituted aryl substituents with radioactive iodide is described. The method takes advantage of the ipso‐directing and activating properties of trimethylsilyl substituents on the arenes. The method was demonstrated on a griseoful...

  6. Validity of the remote food photography method against doubly labeled water among minority preschoolers

    Science.gov (United States)

    The aim of this study was to determine the validity of energy intake (EI) estimations made using the remote food photography method (RFPM) compared to the doubly labeled water (DLW) method in minority preschool children in a free-living environment. Seven days of food intake and spot urine samples...

  7. A new method for tritium labelling of neuraminidase from Vibrio cholerae

    International Nuclear Information System (INIS)

    Keune, D.

    1981-01-01

    This research work related to the radioactive labelling with tritium of the enzyme neuraminidase from Vibrio cholerae by an easily handled method. The reactive compound N-propionyloxysuccinimide, the ester of propionic acid and N-hydroxysuccinimide, offered a suitable labelling reagent. For comparison purposes an already known method of labelling neuraminidase with tritium by the oxidation of hydroxyl groups of the hydrocarbon chain of the enzymal protein and subsequent reduction of the aldehyde groups formed with tritiated sodium borhydride, was also carried out. The advantages and disadvantages of both methods are described in detail, in particular with regard to yields of radioactivity and the influence on enzyme activity. The fact that only 1 mg enzymal protein was available for each modification of the enzyme molecule posed particular problems and, as a consequence, extensive preliminary experiments had to be carried with another protein (beef serum album) in the same concentration range. (orig./MG) [de

  8. Sensory evaluation method of building materials for labelling purposes

    Energy Technology Data Exchange (ETDEWEB)

    Tirkkonen, T.; Saarela, K. [VTT Building and Transport, Espoo (Finland); Kukkonen, E. [Ausum (Finland)

    2004-07-01

    This thesis presents a distributed architecture for the use of wireless sensors in the management of electrical distribution systems. Although the general concept of using wireless sensors for measuring quantities of power lines has previously been introduced, the proposed solutions have not been well integrated within the power system equipment and the automation system. This has severely compromised the applicability of the sensors in the field. However, wireless sensors have several features that make them an attractive instrumentation solution in the harsh environment of electrical distribution networks. Wireless sensors do not need signal or power cables and they are therefore easy to install and use in system refurbishment. They thus provide an interesting and cost effective alternative that is worth studying. In this thesis, the concept developed for enhancing the applicability and integration of wireless sensors within the power distribution system is based on an architecture that uses the properties of the system itself to preserve functionality and to minimise the power consumption and thus the size and weight of the wireless sensors. In this architecture the following essential components and aspects are identified. First, a time synchronisation method for wireless sensors is developed. The main goal with this method is to provide energy optimised means to determine the phase angle in wireless sensor cells where several sensors measure phase current and one sensor measures voltage. Secondly, a fault detection and location method with wireless sensors is proposed. This method enables implementation of fault management using wireless sensors that operate autonomously and asynchronously and measure only phase current. With this approach the hardware and software implementation of a sensor can be simplified compared to previous solutions. The combination of these developed methods enables the construction of lighter, smaller and more flexible wireless

  9. Safe genetic modification of cardiac stem cells using a site-specific integration technique.

    Science.gov (United States)

    Lan, Feng; Liu, Junwei; Narsinh, Kazim H; Hu, Shijun; Han, Leng; Lee, Andrew S; Karow, Marisa; Nguyen, Patricia K; Nag, Divya; Calos, Michele P; Robbins, Robert C; Wu, Joseph C

    2012-09-11

    Human cardiac progenitor cells (hCPCs) are a promising cell source for regenerative repair after myocardial infarction. Exploitation of their full therapeutic potential may require stable genetic modification of the cells ex vivo. Safe genetic engineering of stem cells, using facile methods for site-specific integration of transgenes into known genomic contexts, would significantly enhance the overall safety and efficacy of cellular therapy in a variety of clinical contexts. We used the phiC31 site-specific recombinase to achieve targeted integration of a triple fusion reporter gene into a known chromosomal context in hCPCs and human endothelial cells. Stable expression of the reporter gene from its unique chromosomal integration site resulted in no discernible genomic instability or adverse changes in cell phenotype. Namely, phiC31-modified hCPCs were unchanged in their differentiation propensity, cellular proliferative rate, and global gene expression profile when compared with unaltered control hCPCs. Expression of the triple fusion reporter gene enabled multimodal assessment of cell fate in vitro and in vivo using fluorescence microscopy, bioluminescence imaging, and positron emission tomography. Intramyocardial transplantation of genetically modified hCPCs resulted in significant improvement in myocardial function 2 weeks after cell delivery, as assessed by echocardiography (P=0.002) and MRI (P=0.001). We also demonstrated the feasibility and therapeutic efficacy of genetically modifying differentiated human endothelial cells, which enhanced hind limb perfusion (Pmodification system is a safe, efficient tool to enable site-specific integration of reporter transgenes in progenitor and differentiated cell types.

  10. Tub-Tag Labeling; Chemoenzymatic Incorporation of Unnatural Amino Acids.

    Science.gov (United States)

    Helma, Jonas; Leonhardt, Heinrich; Hackenberger, Christian P R; Schumacher, Dominik

    2018-01-01

    Tub-tag labeling is a chemoenzymatic method that enables the site-specific labeling of proteins. Here, the natural enzyme tubulin tyrosine ligase incorporates noncanonical tyrosine derivatives to the terminal carboxylic acid of proteins containing a 14-amino acid recognition sequence called Tub-tag. The tyrosine derivative carries a unique chemical reporter allowing for a subsequent bioorthogonal modification of proteins with a great variety of probes. Here, we describe the Tub-tag protein modification protocol in detail and explain its utilization to generate labeled proteins for advanced applications in cell biology, imaging, and diagnostics.

  11. The method of determination of micro quantities of labeled iodide in carrier free Na125 solution

    International Nuclear Information System (INIS)

    Kholbaev, A.Kh.; Shilin, E.A.

    1996-01-01

    The method of determination of microquantities of labelled iodide in Na 125 carrier-free solution was elaborated. This method permits to increase the sensitivity and radiation protection of the determination of labeled iodide. It includes oxidation of iodide by iodate in diluted sulphuric acid with molar concentration 0,03-0,04 mole/l. The extraction of I 2 is made by toluene. The coloured solution is made and optical density is measured at λ=640 nm at the 10 mm optical path .(A.A.D.)

  12. Hemoglobin radiolabeling: in vitro and in vivo comparison of iodine labeling with iodogen and a new method for technetium labeling

    International Nuclear Information System (INIS)

    Bleeker, W.K.; Feitsma, R.I.J.; Pauwels, E.K.J.; Plas, J. van der; Agterberg, J.; Rigter, G.; Bakker, J.C.

    1989-01-01

    The present investigation compares the suitability of two radiolabeling techniques for hemoglobin. 125 I labeling of hemoglobin with Iodogen as iodinating agent caused major changes in the chromatographic behaviour and an accelerated plasma clearance of the labeled hemoglobin in rats. A recently developed two-step procedure for 99m Tc labeling gave better results. The label had only minimal influence on the chromatographic behaviour of hemoglobin. In vivo, no free label occurred in the circulation and no transfer of the label to other plasma proteins took place. The plasma clearance of 99m Tc-labeled hemoglobin in rats was slowed. However, this could be explained entirely by diminishing glomerular filtration, probably by inhibition of the dissociation of the hemoglobin molecule into dimers. The plasma clearance of hemoglobin modified by intramolecular cross-linking, which prevents dissociation of the molecule into dimers and thus excretion by the kidney, was not influenced by the label. We conclude that the 99m Tc labeling procedure is suitable for in vivo distribution studies of hemoglobin when it is taken into account that the urinary excretion is underestimated. For cross-linked hemoglobin, which is more promising as plasma expander, no such restriction exists. (author)

  13. Site specific incorporation of keto amino acids into proteins

    Science.gov (United States)

    Schultz, Peter G [La Jolla, CA; Wang, Lei [San Diego, CA

    2008-10-07

    Compositions and methods of producing components of protein biosynthetic machinery that include orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, and orthogonal pairs of tRNAs/synthetases, which incorporate keto amino acids into proteins are provided. Methods for identifying these orthogonal pairs are also provided along with methods of producing proteins with keto amino acids using these orthogonal pairs.

  14. Site-specific mouth rinsing can improve oral odor by altering bacterial counts. Blind crossover clinical study.

    Science.gov (United States)

    Alqumber, Mohammed A; Arafa, Khaled A

    2014-11-01

    To determine whether site-specific mouth rinsing with oral disinfectants can improve oral odor beyond the traditional panoral mouth disinfection with mouth rinses by targeting specifically oral malodor implicated anaerobic bacteria. Twenty healthy fasting subjects volunteered for a blinded prospective, descriptive correlational crossover cross-section clinical trial conducted during the month of Ramadan between July and August 2013 in Albaha province in Saudi Arabia involving the application of Listerine Cool Mint mouth rinse by either the traditional panoral rinsing method, or a site-specific disinfection method targeting the subgingival and supragingival plaque and the posterior third of the tongue dorsum, while avoiding the remaining locations within the oral cavity. The viable anaerobic and aerobic bacterial counts, volatile sulfur compounds (VSCs) levels, organoleptic assessment of oral odor, and the tongue-coating index were compared at baseline, one, 5, and 9 hours after the treatment. The site-specific disinfection method reduced the VSCs and anaerobic bacterial loads while keeping the aerobic bacterial numbers higher than the traditional panoral rinsing method. Site-specific disinfection can more effectively maintain a healthy oral cavity by predominantly disinfecting the niches of anaerobic bacteria within the oral cavity.

  15. Development of methods of labeling pentavalent DMSA with 99mTc and 188Re

    International Nuclear Information System (INIS)

    Brambilla, Tania de Paula

    2009-01-01

    Technetium-99 m is the most useful radionuclide in diagnostic imaging procedures in Nuclear Medicine, more than 80 percent of radiopharmaceuticals are 99m Tc-labeled compounds. 99m Tc-DMSA(V) has been used for imaging of soft tissue, head and neck tumors. It shows a particularly high specificity for medullary thyroid carcinoma and bone metastases in a variety of cancers. Biodistribution studies of 188 Re-DMSA(V) have shown that its general pharmacokinetic properties are similar to that of 99m Tc-DMSA(V), so this agent could be used for targeted radiotherapy of these tumors. The aim of this work is the development of methods of labeling DMSA(V) with 99m Tc and 188 Re. 99m Tc-DMSA(V) can be prepared by two methods. One of them is the indirect one, through the use of a commercial kit of DMSA (III), by adjusting the pH from 2.5 to ∼ 8.5 with NaHCO 3 . This method was evaluated and optimized presenting high labeling yields. The other method is the direct one, through the preparation of a lyophilised kit ready for labeling with 99m Tc, being the method of interest of this work, due to the easy of its clinical use. The most adequate formulation of the kit was: 1.71 mg of DMSA, 0.53 mg of SnCl 2 .2H 2 O and 0.83 mg of ascorbic acid (pH 9). Labeling yields higher than 95% were achieved labeling this kit with 1 to 2 m L of 99m Tc with activities up to 4736 MBq (128 mCi). The kit was stable up to 6 months and biodistribution studies confirmed the quality of the DMSA (V) labeled with 99m Tc using this kit. The reduction potential of Re is lower than the one for Tc, so the labeling conditions of 188 Re-DMSA(V) are different from the ones used for 99m Tc- DMSA(V). 188 Re-DMSA(V) is prepared in acid solution, that makes it possible to use the DMSA (III) commercial kit developed for labeling with 99m Tc, prepared in pH 2.5, for labeling with 188 Re. Labeling yields higher than 95% were achieved with this methodology, with a rection time of 30 minutes at 100 deg C using no more

  16. Assessment of inflammatory bowel disease with two different 99mTc-leucocytes labelling methods

    International Nuclear Information System (INIS)

    Cardoso, V.N.; Plaza, P.J.L.; Roca, M.; Armero, F.; Martin-Comin, J.

    2002-01-01

    Aim of this study was to retrospectively compare the diagnostic accuracy of 99mTc-HMPAO white blood cell scintigraphy using two different cell suspension mediums: leukocyte poor plasma (LPP) and Hanks' Balanced Salt Solution (HBSS) in patients with suspicion of active inflammatory bowel disease. Materials and Methods: Leukocytes from 30 patients were labelled using LPP and in 28 using HBSS . In LPP method the leukocytes were resuspended in 0,5 ml cell-free plasma while in HBSS method the cells were resuspended in 0,5 ml HBSS. Scintigraphic images were obtained at 30 min and 2 h after injection of 185-200 MBq 99mTc-HMPAO leukocytes. Results: Leukocytes labelling efficiency were 65,5%, and 89,0%, respectively for LPP and HBSS methods. There were 22 true-positive, 7 true-negative and 1 false-negative results in the LPP group, while in the HBSS group results were 18, 10 and 0, respectively. Diagnostic accuracy was similar with both methods though sensitivity was slightly higher in the HBSS group. Conclusion These date indicate that leukocytes scintigraphy labelled using HBSS as resuspension medium should be used as first option method for WBC labelling and diagnosis of inflammatory bowel disease

  17. Site-specific photoconjugation of antibodies using chemically synthesized IgG-binding domains.

    Science.gov (United States)

    Perols, Anna; Karlström, Amelie Eriksson

    2014-03-19

    Site-specific labeling of antibodies can be performed using the immunoglobulin-binding Z domain, derived from staphylococcal protein A (SpA), which has a well-characterized binding site in the Fc region of antibodies. By introducing a photoactivable probe in the Z domain, a covalent bond can be formed between the Z domain and the antibody by irradiation with UV light. The aim of this study was to improve the conjugation yield for labeling of different subclasses of IgG having different sequence composition, using a photoactivated Z domain variant. Four different variants of the Z domain (Z5BPA, Z5BBA, Z32BPA, and Z32BBA) were synthesized to investigate the influence of the position of the photoactivable probe and the presence of a flexible linker between the probe and the protein. For two of the variants, the photoreactive benzophenone group was introduced as part of an amino acid side chain by incorporation of the unnatural amino acid benzoylphenylalanine (BPA) during peptide synthesis. For the other two variants, the photoreactive benzophenone group was attached via a flexible linker by coupling of benzoylbenzoic acid (BBA) to the ε-amino group of a selectively deprotected lysine residue. Photoconjugation experiments using human IgG1, mouse IgG1, and mouse IgG2A demonstrated efficient conjugation for all antibodies. It was shown that differences in linker length had a large impact on the conjugation efficiency for labeling of mouse IgG1, whereas the positioning of the photoactivable probe in the sequence of the protein had a larger effect for mouse IgG2A. Conjugation to human IgG1 was only to a minor extent affected by position or linker length. For each subclass of antibody, the best variant tested using a standard conjugation protocol resulted in conjugation efficiencies of 41-66%, which corresponds to on average approximately one Z domain attached to each antibody. As a combination of the two best performing variants, Z5BBA and Z32BPA, a Z domain variant with

  18. A novel facile method of labeling octreotide with (18)F-fluorine.

    Science.gov (United States)

    Laverman, Peter; McBride, William J; Sharkey, Robert M; Eek, Annemarie; Joosten, Lieke; Oyen, Wim J G; Goldenberg, David M; Boerman, Otto C

    2010-03-01

    Several methods have been developed to label peptides with (18)F. However, in general these are laborious and require a multistep synthesis. We present a facile method based on the chelation of (18)F-aluminum fluoride (Al(18)F) by 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA). The method is characterized by the labeling of NOTA-octreotide (NOTA-d-Phe-cyclo[Cys-Phe-d-Trp-Lys-Thr-Cys]-Throl (MH(+) 1305) [IMP466]) with (18)F. Octreotide was conjugated with the NOTA chelate and labeled with (18)F in a 2-step, 1-pot method. The labeling procedure was optimized with regard to the labeling buffer, peptide, and aluminum concentration. Radiochemical yield, specific activity, in vitro stability, and receptor affinity were determined. Biodistribution of (18)F-IMP466 was studied in AR42J tumor-bearing mice and compared with that of (68)Ga-labeled IMP466. In addition, small-animal PET/CT images were acquired. IMP466 was labeled with Al(18)F in a single step with 50% yield. The labeled product was purified by high-performance liquid chromatography to remove unbound Al(18)F and unlabeled peptide. The radiolabeling, including purification, was performed in 45 min. The specific activity was 45,000 GBq/mmol, and the peptide was stable in serum for 4 h at 37 degrees C. Labeling was performed at pH 4.1 in sodium citrate, sodium acetate, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, and 2-(N-morpholino)ethanesulfonic acid buffer and was optimal in sodium acetate buffer. The apparent 50% inhibitory concentration of the (19)F-labeled IMP466 determined on AR42J cells was 3.6 nM. Biodistribution studies at 2 h after injection showed a high tumor uptake of (18)F-IMP466 (28.3 +/- 5.2 percentage injected dose per gram [%ID/g]; tumor-to-blood ratio, 300 +/- 90), which could be blocked by an excess of unlabeled peptide (8.6 +/- 0.7 %ID/g), indicating that the accumulation in the tumor was receptor-mediated. Biodistribution of (68)Ga-IMP466 was similar to that of (18)F-IMP466. (18)F

  19. Site-specific accumulation and dynamic change of flavonoids in Apocyni Veneti Folium.

    Science.gov (United States)

    Chen, Cui-Hua; Xu, Hu; Liu, Xun-Hong; Zou, Li-Si; Wang, Mei; Liu, Zi-Xiu; Fu, Xing-Sheng; Zhao, Hui; Yan, Ying

    2017-12-01

    Site-specific accumulation of flavonoids in Apocyni Veneti Folium was determined by laser scanning confocal microscope (LSCM) and the localization of catechins also was observed via vanillin-HCl staining under the conventional optical microscope. The contents of five flavonoids in Apocyni Veneti Folium from different harvest times and growth parts were measured using HPLC method. LSCM observation showed that flavonoids are accumulated in cuticle of epidermal cells and vessel walls, especially in protoplasts and nucleolus of the collenchyma cells and the epidermal cells. Catechins are localized in the palisade parenchyma cells and vessel walls, particularly in the laticifers found in the phloem. On the basis of the difference of the maximal emission wavelength between quercetin and kaempferol derivatives which have fluorescence behavior by appropriate treatment, kaempferol and its derivatives are localized exclusively in the cuticle. Results showed that the content of astragalin in Apocyni Veneti Folium from different parts revealed the decreasing trend, while hyperin and isoquercitrin were higher in June and July analyzed by HPLC. In summary, the site-specific accumulation of flavonoids in Apocyni Veneti Folium can be determined by LSCM and vanillin-HCl staining. The contents of flavonoids in Apocyni Veneti Folium are correlated with harvest times and growth parts. © 2017 Wiley Periodicals, Inc.

  20. Determining site-specific background level with geostatistics for remediation of heavy metals in neighborhood soils

    Directory of Open Access Journals (Sweden)

    Tammy M. Milillo

    2017-03-01

    Full Text Available The choice of a relevant, uncontaminated site for the determination of site-specific background concentrations for pollutants is critical for planning remediation of a contaminated site. The guidelines used to arrive at concentration levels vary from state to state, complicating this process. The residential neighborhood of Hickory Woods in Buffalo, NY is an area where heavy metal concentrations and spatial distributions were measured to plan remediation. A novel geostatistics based decision making framework that relies on maps generated from indicator kriging (IK and indicator co-kriging (ICK of samples from the contaminated site itself is shown to be a viable alternative to the traditional method of choosing a reference site for remediation planning. GIS based IK and ICK, and map based analysis are performed on lead and arsenic surface and subsurface datasets to determine site-specific background concentration levels were determined to be 50 μg/g for lead and 10 μg/g for arsenic. With these results, a remediation plan was proposed which identified regions of interest and maps were created to effectively communicate the results to the environmental agencies, residents and other interested parties.

  1. New developments for the site-specific attachment of protein to surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Camarero, J A

    2005-05-12

    Protein immobilization on surfaces is of great importance in numerous applications in biology and biophysics. The key for the success of all these applications relies on the immobilization technique employed to attach the protein to the corresponding surface. Protein immobilization can be based on covalent or noncovalent interaction of the molecule with the surface. Noncovalent interactions include hydrophobic interactions, hydrogen bonding, van der Waals forces, electrostatic forces, or physical adsorption. However, since these interactions are weak, the molecules can get denatured or dislodged, thus causing loss of signal. They also result in random attachment of the protein to the surface. Site-specific covalent attachment of proteins onto surfaces, on the other hand, leads to molecules being arranged in a definite, orderly fashion and uses spacers and linkers to help minimize steric hindrances between the protein surface. This work reviews in detail some of the methods most commonly used as well as the latest developments for the site-specific covalent attachment of protein to solid surfaces.

  2. Site-Specific Incorporation of Functional Components into RNA by an Unnatural Base Pair Transcription System

    Directory of Open Access Journals (Sweden)

    Rie Kawai

    2012-03-01

    Full Text Available Toward the expansion of the genetic alphabet, an unnatural base pair between 7-(2-thienylimidazo[4,5-b]pyridine (Ds and pyrrole-2-carbaldehyde (Pa functions as a third base pair in replication and transcription, and provides a useful tool for the site-specific, enzymatic incorporation of functional components into nucleic acids. We have synthesized several modified-Pa substrates, such as alkylamino-, biotin-, TAMRA-, FAM-, and digoxigenin-linked PaTPs, and examined their transcription by T7 RNA polymerase using Ds-containing DNA templates with various sequences. The Pa substrates modified with relatively small functional groups, such as alkylamino and biotin, were efficiently incorporated into RNA transcripts at the internal positions, except for those less than 10 bases from the 3′-terminus. We found that the efficient incorporation into a position close to the 3′-terminus of a transcript depended on the natural base contexts neighboring the unnatural base, and that pyrimidine-Ds-pyrimidine sequences in templates were generally favorable, relative to purine-Ds-purine sequences. The unnatural base pair transcription system provides a method for the site-specific functionalization of large RNA molecules.

  3. Site-specific confocal fluorescence imaging of biological microstructures in a turbid medium

    International Nuclear Information System (INIS)

    Saloma, Caesar; Palmes-Saloma, Cynthia; Kondoh, Hisato

    1998-01-01

    Normally transparent biological structures in a turbid medium are imaged using a laser confocal microscope and multiwavelength site-specific fluorescence labelling. The spatial filtering capability of the detector pinhole in the confocal microscope limits the number of scattered fluorescent photons that reach the photodetector. Simultaneous application of different fluorescent markers on the same sample site minimizes photobleaching by reducing the excitation time for each marker. A high-contrast grey-level image is also produced by summing confocal images of the same site taken at different fluorescence wavelengths. Monte Carlo simulations are performed to obtain the quantitative behaviour of confocal fluorescence imaging in turbid media. Confocal images of the following samples were also obtained: (i) 15 μm diameter fluorescent spheres placed 1.16 mm deep beneath an aqueous suspension of 0.0823 μm diameter polystyrene latex spheres, and (ii) hindbrain of a whole-mount mouse embryo (age 10 days) that was stained to fluoresce at 515 nm and 580 nm peak wavelengths. Expression of RNA transcripts of a gene within the embryo hindbrain was detected by a fluorescence-based whole-mount in situ hybridization procedure that we recently tested. (author)

  4. Preparation of tritium- or deuterium-labeled vitamin D analogs by a convenient general method

    International Nuclear Information System (INIS)

    Paaren, H.E.; Fivizzani, M.A.; Schnoes, H.K.; DeLuca, H.F.

    1981-01-01

    The three-step conversion of vitamin D analogs to 6-oxo-3,5-cyclovitamin D derivatives followed by reduction with a tritide or deuteride reagent and subsequent cycloreversion gives 6-tritio(deutero)vitamin D derivatives and corresponding 5,6-trans-analogs. The method is general and affords the 6-labeled-vitamin D analogs in approx. =20% overall yield

  5. Study on labelling methods of 125I-RC-160 and its biodistribution in animals

    International Nuclear Information System (INIS)

    Wang Jing; Wang Xiqing; Wang Liangang; Li Fujun; Deng Jinglan

    2002-01-01

    A method for the iodination of peptide RC-160 with high efficiency was developed. RC-160 was iodinated with N-bromosuccinimide (NBS) as oxidant, the conventional chloramine T (Ch-T) method was used as control. The labelling condition of NBS method was optimized and radiolabelled conjugate 125 I-RC-160 was assessed as follows: no further purification was needed, the measured labelling yield of 125 I-RC-160 was 92% and the specific activity was 1.95 x 10 12 Bq/m mol. The yield increased as the amount of NBS increased. The optimal ratio of RC-160 (μg): 125 I (MBq): NBS(μg) was 3:7.4:1. For Ch-T method, the labelling yield is 56% and specific activity was 0.65 x 10 12 Bq/m mol; but after purification by SepPak-C 18 , the labelling yield may reach as high as 92%. 1h after injection, radioactivity in blood decreased by 87.2%. No obvious concentration of 125 I-RC-160 in thyroid or kidney was observed

  6. Double-label autoradiographic deoxyglucose method for sequential measurement of regional cerebral glucose utilization

    Energy Technology Data Exchange (ETDEWEB)

    Redies, C; Diksic, M; Evans, A C; Gjedde, A; Yamamoto, Y L

    1987-08-01

    A new double-label autoradiographic glucose analog method for the sequential measurement of altered regional cerebral metabolic rates for glucose in the same animal is presented. This method is based on the sequential injection of two boluses of glucose tracer labeled with two different isotopes (short-lived /sup 18/F and long-lived /sup 3/H, respectively). An operational equation is derived which allows the determination of glucose utilization for the time period before the injection of the second tracer; this equation corrects for accumulation and loss of the first tracer from the metabolic pool occurring after the injection of the second tracer. An error analysis of this operational equation is performed. The double-label deoxyglucose method is validated in the primary somatosensory (''barrel'') cortex of the anesthetized rat. Two different rows of whiskers were stimulated sequentially in each rat; the two periods of stimulation were each preceded by an injection of glucose tracer. After decapitation, dried brain slices were first exposed, in direct contact, to standard X-ray film and then to uncoated, ''tritium-sensitive'' film. Results show that the double-label deoxyglucose method proposed in this paper allows the quantification and complete separation of glucose utilization patterns elicited by two different stimulations sequentially applied in the same animal.

  7. Tritium labelling and characterization of the antimalarial drug (+/-)-chloroquine by several methods

    Energy Technology Data Exchange (ETDEWEB)

    Egan, J.A.Judith A.; Laseter, Anne G; Filer, C.N.Crist N. E-mail: crist.filer@perkinelmer.com

    2002-09-01

    To study its mechanism of antimalarial action, a tritium labelled analogue of (+/-)-chloroquine was required at high specific activity. Two synthetic methods were successfully employed. [3-{sup 3}H] (+/-)-Chloroquine 2 was prepared by the catalytic tritium dehalogenation of an iodo precursor and [N-ethyl-{sup 3}H] (+/-)-chloroquine 4 was synthesized by the alkylation of (+/-)-desethylchloroquine with [{sup 3}H] ethyl iodide.

  8. Tritium labelling and characterization of the antimalarial drug (+/-)-chloroquine by several methods

    International Nuclear Information System (INIS)

    Egan, J.A.Judith A.; Laseter, Anne G.; Filer, C.N.Crist N.

    2002-01-01

    To study its mechanism of antimalarial action, a tritium labelled analogue of (+/-)-chloroquine was required at high specific activity. Two synthetic methods were successfully employed. [3- 3 H] (+/-)-Chloroquine 2 was prepared by the catalytic tritium dehalogenation of an iodo precursor and [N-ethyl- 3 H] (+/-)-chloroquine 4 was synthesized by the alkylation of (+/-)-desethylchloroquine with [ 3 H] ethyl iodide

  9. Physical activity assessment : comparison between movement registration and doubly labeled water method

    NARCIS (Netherlands)

    Westerterp, K.; Bouten, C.V.C.

    1997-01-01

    The doubly labeled water method for the measurement of average daily metabolic rate (ADMR), combined with a measurement of resting metabolic rate, permits the calculation of energy expenditure for physical activity under normal daily living conditions. This procedure was used to evaluate the use of

  10. Technetium-99m labeled radiodiagnostic agents for liver and bone marrow scanning and method of preparation

    International Nuclear Information System (INIS)

    Molinski, V.J.; Peacock, F.R.

    1977-01-01

    An improved technetium-99m labeled colloid and method of preparation comprising reducing technetium-99m with stannous oxalate and stabilizing with sodium phytate are described. This radiodiagnostic agent is useful in the scintigraphic examination of the reticuloendothelial system, particularly the liver. In addition, by autoclaving this product with saline, it becomes a superior bone marrow scanning agent

  11. A comparison of labelled antibody methods for the detection of virus antigens in cell monolayers

    International Nuclear Information System (INIS)

    Oram, J.D.; Crooks, A.J.

    1979-01-01

    A number of labelled antibody methods have been applied to the detection of Semliki Forest virus antigens after replication of the virus in monolayers of host cells in multi-well polystyrene plates. The importance of several reaction variables has been investigated and the sensitivity of the methods compared for different periods of virus replication. Direct assays with radio-labelled antibody (RLA) and indirect assays peroxidase-antiperoxidase complexes (PAP) were equally sensitive. Direct and indirect assays using enzyme-linked antibodies (ELA) were slightly less sensitive than the direct RLA and PAP methods but were more sensitive than the indirect RLA or fluorescent antibody (FLA) methods. Direct assays using ELA were more rapid and easier to perform than the other assay methods. (Auth.)

  12. Site-specific fab fragment biotinylation at the conserved nucleotide binding site for enhanced Ebola detection.

    Science.gov (United States)

    Mustafaoglu, Nur; Alves, Nathan J; Bilgicer, Basar

    2015-07-01

    The nucleotide binding site (NBS) is a highly conserved region between the variable light and heavy chains at the Fab domains of all antibodies, and a small molecule that we identified, indole-3-butyric acid (IBA), binds specifically to this site. Fab fragment, with its small size and simple production methods compared to intact antibody, is good candidate for use in miniaturized diagnostic devices and targeted therapeutic applications. However, commonly used modification techniques are not well suited for Fab fragments as they are often more delicate than intact antibodies. Fab fragments are of particular interest for sensor surface functionalization but immobilization results in damage to the antigen binding site and greatly reduced activity due to their truncated size that allows only a small area that can bind to surfaces without impeding antigen binding. In this study, we describe an NBS-UV photocrosslinking functionalization method (UV-NBS(Biotin) in which a Fab fragment is site-specifically biotinylated with an IBA-EG11-Biotin linker via UV energy exposure (1 J/cm(2)) without affecting its antigen binding activity. This study demonstrates successful immobilization of biotinylated Ebola detecting Fab fragment (KZ52 Fab fragment) via the UV-NBS(Biotin) method yielding 1031-fold and 2-fold better antigen detection sensitivity compared to commonly used immobilization methods: direct physical adsorption and NHS-Biotin functionalization, respectively. Utilization of the UV-NBS(Biotin) method for site-specific conjugation to Fab fragment represents a proof of concept use of Fab fragment for various diagnostic and therapeutic applications with numerous fluorescent probes, affinity molecules and peptides. © 2015 Wiley Periodicals, Inc.

  13. Preparation of a pure 99mTc-F(ab')2 radioimmunoconjugate by direct labeling methods

    International Nuclear Information System (INIS)

    Griffiths, G.L.; Jones, A.L.; Hansen, H.J.; Goldenberg, D.M.

    1994-01-01

    Intact IgG and Fab' can be labeled directly with 99m Tc to give quantitative incorporation of radioactivity into the protein. With F(ab') 2 the reductive conditions yield a mixture of 99m Tc-F(ab') 2 and 99m Tc-Fab'. We now report a direct labeling method to produce only 99m Tc-F(ab') 2 in quantitative yield and contaminated with 99m Tc-Fab'. The properties, stability and biodistribution of the 99m Tc-F(ab') 2 have been compared to 99m Tc-Fab'. This new technology will allow us to compare technetium direct-labeled IgG, F(ab') 2 and Fab' derivatives of the same antibody for radioimmunodetection. (author)

  14. Biosynthetic incorporation of [75Se]selenomethionine: a new method for labelling lymphocyte membrane antigens

    International Nuclear Information System (INIS)

    Dosseto, M.; Rohner, C.; Pierres, M.; Goridis, C.

    1981-01-01

    A novel approach for radiolabelling lymphocyte membrane antigens is described. This technique is based on the use of the γ-emitting amino acid analogue [ 75 Se]selenomethionine. Human HLA-A, B, C and DR heavy and light chains and mouse Ia antigens were efficiently labelled by this technique and were precipitated with monoclonal antibodies. Approximately the same radioactivity was incorporated into the HLA-A, B, C chains whether [ 75 Se]selenomethionine, [ 35 S]methionine or [ 3 H]leucine were used as precursors. Easily detectable as a γ-emitter, [ 75 Se]selenomethionine thus constitutes a useful biosynthetic label of lymphocyte surface antigens. The same method was used to label immunoglobulins produced by hybridomas and to determine the nature of the secreted light chains. (Auth.)

  15. A novel colloidal gold labeled antigen for the detection of Deoxynivalenol using an immunochromatographic assay method

    Science.gov (United States)

    Jin, Yu; Liu, Renrong; Zhu, Lixin; Chen, Zhenzhen

    2017-11-01

    In this paper, an immunochromatographic assay card was developed for the detection of DON in feed and cereals using a novel colloidal gold labeling method. For the colloidal gold immunochromatographic rapid detection (GICD) card, a monoclonal antibody DON-mAb and a goat anti-chicken IgY were drawn on NC membrane as the test line (T line) and the control line (C line) respectively. A gold labeled DON-CBSA conjugate and a gold labeled chicken IgY were sprayed onto the conjugate pad. The GICD card has cut-off levels of 50ng/mL for DON, which is invulnerable to matrix interference, and applicable to a wide range of samples. The GICD detecting results of feed and grain samples were compared with the results of ELISA testing, which showed good consistency.

  16. A general method for tritium labelling of benzimidazole carbamates by catalytic exchange in dioxane solutions

    Energy Technology Data Exchange (ETDEWEB)

    Lacey, E [Commonwealth Scientific and Industrial Research Organization, Glebe, NSW (Australia). Div. of Animal Health, McMaster Lab.; Dawson, M [Sydney Univ. (Australia). Dept. of Pharmacy; Long, M A; Than, C [New South Wales Univ., Kensington (Australia). School of Chemistry

    1989-12-01

    Benzimidazole carbamates (BZCs) act as inhibitors of the tubulin-microtubule equilibria in eukaryotic organisms. Recently drug resistance to this class of compounds in helminth parasites has been shown to be due to a reduced ability of resistant tubulin to bind BZCs. In order to quantitate the nature of the tubulin-BZC interaction a general method for the specific tritium labelling of BZCs has been developed. The BZCs: mebendazole, oxfendazole, parbendazole, oxibendazole, albendazole and fenbendazole were labelled by catalytic exchange using palladium on calcium carbonate in pure dioxane at 60{sup 0}C under tritium gas. The position of label incorporation for tritiated albendazole was determined by tritium-NMR as the 4-position of benzimadazole nucleus. The yields for individual BZCs varied from 8 to 68% for a range of specific activity of 0.44 to 13.4 Ci/mmole. (author).

  17. A general method for tritium labelling of benzimidazole carbamates by catalytic exchange in dioxane solutions

    International Nuclear Information System (INIS)

    Lacey, E.; Dawson, M.; Long, M.A.; Than, C.

    1989-01-01

    Benzimidazole carbamates (BZCs) act as inhibitors of the tubulin-microtubule equilibria in eukaryotic organisms. Recently drug resistance to this class of compounds in helminth parasites has been shown to be due to a reduced ability of resistant tubulin to bind BZCs. In order to quantitate the nature of the tubulin-BZC interaction a general method for the specific tritium labelling of BZCs has been developed. The BZCs: mebendazole, oxfendazole, parbendazole, oxibendazole, albendazole and fenbendazole were labelled by catalytic exchange using palladium on calcium carbonate in pure dioxane at 60 0 C under tritium gas. The position of label incorporation for tritiated albendazole was determined by tritium-NMR as the 4-position of benzimadazole nucleus. The yields for individual BZCs varied from 8 to 68% for a range of specific activity of 0.44 to 13.4 Ci/mmole. (author)

  18. Practical Application of Site-Specific Earthquake Early Warning (EEW) System

    International Nuclear Information System (INIS)

    Kanda, Katsuhisa

    2014-01-01

    The development of an on-site warning system was reported. This system improves the timing of warnings and reduces the number of false alarms by improving the method of estimating the JMA seismic intensity using earthquake early warning system information based on site-specific data. Moreover, the development of an application for practical use in a construction company and an integrated system for realizing system shutdown was also reported. The concept of this system is based on the following. Seismic intensity is not distributed concentrically, and the attenuation relationship cannot explain the distribution of seismic intensity precisely. The standard method of seismic intensity prediction is construed as 'attenuation relationship + soil amplification factor', but this may be improved in the reformulation 'original attenuation relationship for each site + correction factors dependent on the epicenter location and depth' using a seismic intensity database that includes data on recent and historical earthquakes. (authors)

  19. Carbohydrates on Proteins: Site-Specific Glycosylation Analysis by Mass Spectrometry

    Science.gov (United States)

    Zhu, Zhikai; Desaire, Heather

    2015-07-01

    Glycosylation on proteins adds complexity and versatility to these biologically vital macromolecules. To unveil the structure-function relationship of glycoproteins, glycopeptide-centric analysis using mass spectrometry (MS) has become a method of choice because the glycan is preserved on the glycosylation site and site-specific glycosylation profiles of proteins can be readily determined. However, glycopeptide analysis is still challenging given that glycopeptides are usually low in abundance and relatively difficult to detect and the resulting data require expertise to analyze. Viewing the urgent need to address these challenges, emerging methods and techniques are being developed with the goal of analyzing glycopeptides in a sensitive, comprehensive, and high-throughput manner. In this review, we discuss recent advances in glycoprotein and glycopeptide analysis, with topics covering sample preparation, analytical separation, MS and tandem MS techniques, as well as data interpretation and automation.

  20. Molecular characterization of a complex site-specific radiation-induced DNA double-strand break

    International Nuclear Information System (INIS)

    Datta, K.; Dizdaroglu, M.; Jaruga, P.; Neumann, R.D.; Winters, T.A.

    2003-01-01

    Radiation lethality is a function of radiation-induced DNA double-strand breaks (DSB). Current models propose the lethality of a DSB to be a function of its structural complexity. We present here for the first time a map of damage associated with a site-specific double-strand break produced by decay of 125 I in a plasmid bound by a 125 I-labeled triplex forming oligonucleotide ( 125 I-TFO). The E. coli DNA repair enzymes, endonuclease IV (endo IV), endonuclease III (endo III), and formamidopyrimidine-DNA glycosylase (Fpg), which recognize AP sites, and pyrimidine and purine base damage respectively, were used as probes in this study. 125 I-TFO bound plasmid was incubated with and without DMSO at -80 deg C for 1 month. No significant difference in DSB yield was observed under these conditions. A 32 base pair fragment from the upstream side of the decay site was isolated by restriction digestion and enzymatically probed to identify damage sites. Endo IV treatment of the 5'-end labeled upper strand indicated clustering of AP sites within 3 bases downstream and 7 bases upstream of the targeted base. Also, repeated experiments consistently detected an AP site 4 bases upstream of the 125 Itarget base. This was further supported by complementary results with the 3'-end labeled upper strand. Endo IV analysis of the lower strand also shows clustering of AP sites near the DSB end. Endo III and Fpg probing demonstrated that base damage is also clustered near the targeted break site. DSBs produced in the absence of DMSO displayed a different pattern of enzyme sensitive damage than those produced in the presence of DMSO. Identification of specific base damage types within the restriction fragment containing the DSB end was achieved with GC/MS. Base damage consisted of 8-hydroguanine, 8-hydroxyadenine, and 5-hydroxycytosine. These lesions were observed at relative yields of 8-hydroguanine and 5-hydroxycytosine to 8-hydroxyadenine of 7.4:1 and 4.7:1, respectively, in the absence

  1. Method of preparing tritium-labelled thymidine-5'-monophosphates of high specific activity

    International Nuclear Information System (INIS)

    Filip, J.; Vesely, J.; Cihak, A.

    1976-01-01

    A method is described of preparing thymidine-5'-monophosphates labelled with tritium of high specific activity based on enzyme synthesis in vitro. Phosphorylation was carried out using the catalytic effect of an enzyme contained in the supernatant fraction prepared from Yoshida ascites carcinoma in rats. The course of the enzyme reaction can be controlled by the concentration of the individual reaction mixture components. The method described allows obtaining thymidine-5'-monophosphate of radiochemical purity better than 95%. (J.B.)

  2. Site-specific semisynthetic variant of human hemoglobin

    International Nuclear Information System (INIS)

    Hefta, S.A.; Lyle, S.B.; Busch, M.R.; Harris, D.E.; Matthew, J.B.; Gurd, F.R.N.

    1988-01-01

    A single round of Edman degradation was employed to remove the NH 2 -terminal valine from isolated α chains of human hemoglobin. Reconstitution of normal β chains with truncated or substituted α chains was used to form truncated (des-Val 1 -α1) and substituted ([[1- 13 C]Gly 1 ]α1) tetrameric hemoglobin analogs. Structural homology of the analogs with untreated native hemoglobin was established by using several spectroscopic and physical methods. Functional studies indicate that the reconstituted tetrameric protein containing des-Val 1 -α chains has a higher affinity for oxygen, is less influenced by chloride ions or 2,3-biphosphoglycerate, and shows lower cooperativity than native hemoglobin. These results confirm the key functional role of the α-chain NH 2 terminus in mediating cooperative oxygen binding across the dimer interface. The NH 2 -terminal pK/sub 1/2/ value was determined for the [ 13 C]glycine-substituted analog to be 7.46 +/- 0.09 at 15 0 C in the carbon monoxide-liganded form. This value, measured directly by 13 C NMR, agrees with the determination made by the less-direct 13 CO 2 method and confirms the role of this residue as a contributor to the alkaline Bohr effect; however, it is consistent with the presence of an NH 2 -terminal salt bridge to the carboxylate of Arg-141 of the α chain in the liganded form

  3. Indium-111 labeling of leukocytes: a detrimental effect on neutrophil and lymphocyte function and an improved method of cell labelling

    International Nuclear Information System (INIS)

    Segal, A.W.; Deteix, P.; Garcia, R.; Tooth, P.; Zanelli, G.D.; Allison, A.C.

    1978-01-01

    A technique for the labeling of cells with the gamma emitter indium-111 has recently been developed. In this study the effects of the labeling procedure on some in vitro functions of human neutrophils and lymphocytes were investigated. With the standard labeling procedure, neutrophil chemotaxis was reduced to approximately 50% of normal and lymphocytes lost surface receptors and failed to respond to stimulation with phytohemagglutinin. The 8-hydroxyquinoline that is used to chelate the indium is toxic to lymphocytes; accordingly the relationship between the quantity of oxine, the chelation of indium, and cell labeling were investigated. Optimal conditions for In-111 cell labeling were established: 100 million cells in 10 ml Hanks' balanced salt solution are mixed with 5 μg of oxine in a mixture of 50 μl of ethanol and 200 μl of saline; they are incubated at 37 0 C for 10 min and then washed. Initially, neutrophils and lymphocytes appear functionally normal, but after 24 to 48 hr lymphocyte function is impaired as a result of radiation damage. This toxicity may limit studies by external scanning on the distribution and kinetics of lymphocytes labeled with In-111

  4. Increase in the specific radioactivity of tritium-labeled compounds obtained by tritium thermal activation method

    International Nuclear Information System (INIS)

    Badun, G.A.; Chernysheva, M.G.; Ksenofontov, A.L.

    2012-01-01

    A method of tritium introduction into different types of organic molecules that is based on the interaction of atomic tritium with solid organic target is described. Tritium atoms are formed on the hot W-wire, which is heated by the electric current. Such an approach is called 'tritium thermal activation method'. Here we summarize the results of labeling globular proteins (lysozyme, human and bovine serum albumins); derivatives of pantothenic acid and amino acids; ionic surfactants (sodium dodecylsulfate and alkyltrimethylammonium bromides) and nonionic high-molecular weight surfactants - pluronics. For the first time it is observed that if the target-compound is fixed and its radicals are stable the specific radioactivity of the labeled product can be drastically increased (up to 400 times) when the target temperature is ca. 295 K compared with the results obtained at 77 K. The influence of labeling parameters as tritium gas pressure, exposure time and W-wire temperature was tested for each target temperature that results in the optimum labeling conditions with high specific radioactivity and chemical yield of the resulting compound. (orig.)

  5. Does labelling frequency affect N rhizodeposition assessment using the cotton-wick method?

    DEFF Research Database (Denmark)

    Mahieu, S.; Fustec, J.; Jensen, Erik Steen

    2009-01-01

    The aim of the present study was to test and improve the reliability of the 15N cotton-wick method for measuring soil N derived from plant rhizodeposition, a critical value for assessing belowground nitrogen input in field-grown legumes. The effects of the concentration of the 15N labelling...... solution and the feeding frequency on assessment of nitrogen rhizodeposition were studied in two greenhouse experiments using the field pea (Pisum sativum L.). Neither the method nor the feeding frequency altered plant biomass and N partitioning, and the method appeared well adapted for assessing...... the belowground contribution of field-grown legumes to the soil N pool. However, nitrogen rhizodeposition assessment was strongly influenced by the feeding frequency and the concentration of labelling solution. At pod-filling and maturity, despite similar root 15N enrichment, the fraction of plants' belowground...

  6. Site-specific cassette exchange systems in the Aedes aegypti mosquito and the Plutella xylostella moth.

    Directory of Open Access Journals (Sweden)

    Roya Elaine Haghighat-Khah

    Full Text Available Genetically engineered insects are being evaluated as potential tools to decrease the economic and public health burden of mosquitoes and agricultural pest insects. Here we describe a new tool for the reliable and targeted genome manipulation of pest insects for research and field release using recombinase mediated cassette exchange (RMCE mechanisms. We successfully demonstrated the established ΦC31-RMCE method in the yellow fever mosquito, Aedes aegypti, which is the first report of RMCE in mosquitoes. A new variant of this RMCE system, called iRMCE, combines the ΦC31-att integration system and Cre or FLP-mediated excision to remove extraneous sequences introduced as part of the site-specific integration process. Complete iRMCE was achieved in two important insect pests, Aedes aegypti and the diamondback moth, Plutella xylostella, demonstrating the transferability of the system across a wide phylogenetic range of insect pests.

  7. Site-Specific Cassette Exchange Systems in the Aedes aegypti Mosquito and the Plutella xylostella Moth

    Science.gov (United States)

    Haghighat-Khah, Roya Elaine; Scaife, Sarah; Martins, Sara; St John, Oliver; Matzen, Kelly Jean; Morrison, Neil; Alphey, Luke

    2015-01-01

    Genetically engineered insects are being evaluated as potential tools to decrease the economic and public health burden of mosquitoes and agricultural pest insects. Here we describe a new tool for the reliable and targeted genome manipulation of pest insects for research and field release using recombinase mediated cassette exchange (RMCE) mechanisms. We successfully demonstrated the established ΦC31-RMCE method in the yellow fever mosquito, Aedes aegypti, which is the first report of RMCE in mosquitoes. A new variant of this RMCE system, called iRMCE, combines the ΦC31-att integration system and Cre or FLP-mediated excision to remove extraneous sequences introduced as part of the site-specific integration process. Complete iRMCE was achieved in two important insect pests, Aedes aegypti and the diamondback moth, Plutella xylostella, demonstrating the transferability of the system across a wide phylogenetic range of insect pests. PMID:25830287

  8. Nanostructured interfaces with site-specific bioreceptors for immunosensing

    Energy Technology Data Exchange (ETDEWEB)

    Paiva, Telmo O.; Almeida, Inês; Marquês, Joaquim T. [Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, Edifício C8, 1749-016 Lisboa (Portugal); Liu, Wei [NML, Institute of Mechanics, Chinese Academy of Sciences, Beijing, 100190 (China); Institute of Microelectronics, Tsinghua University, Beijing 100084 (China); Niu, Yu [NML, Institute of Mechanics, Chinese Academy of Sciences, Beijing, 100190 (China); Jin, Gang, E-mail: gajin@imech.ac.cn [NML, Institute of Mechanics, Chinese Academy of Sciences, Beijing, 100190 (China); School of Engineering Science, University of Chinese Academy of Sciences, Beijing 100049 (China); Viana, Ana S., E-mail: anaviana@fc.ul.pt [Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, Edifício C8, 1749-016 Lisboa (Portugal)

    2017-08-01

    Highlights: • Innovative and simple strategy to create sensitive immunosensing platforms. • Gold surface modification with dithiocarbamate nanoconjugates of protein A. • CS{sub 2} strongly adsorbed on gold able to block protein nonspecific adsorption. • High performance for antigen detection by properly oriented antibodies. - Abstract: In this work, we propose a simple and effective approach to build nanostructured immunosensor platforms. The one-step strategy relies on i) the in situ formation of dithiocarbamates from the reaction between carbon disulfide and amine groups, present on protein A, ii) their attachment to gold nanoparticles (AuNPs), and iii) the linkage of the modified AuNPs to the electrode surface, which depends on the strong interaction between gold substrates and sulfur moieties. AuNPs and protein A are used to increase the surface coverage of Immunoglobulin G (IgG) and promote the oriented immobilization of the antibodies on the immunosensing interface. The modified gold surfaces with biomolecules were thoroughly characterized by a combination of techniques: UV–vis spectrophotometry, conventional ellipsometry and atomic force microscopy. The immunosensor performance was assessed in real-time, by surface plasmon resonance and by the highly sensitive total internal reflection imaging ellipsometry, through the specific biorecognition between anti-IgG and the immobilized IgG molecules. We demonstrate that the presence of AuNPs improves the sensitivity of the anti-IgG specific detection, whereas the presence of co-adsorbed CS{sub 2} is responsible for blocking the undesired protein nonspecific adsorption to the gold substrate. Overall, we report a simple and innovative one-step method, to chemically modify gold surfaces with protein A and AuNPs, able to specifically detect antigen/antibody interactions with capability of preventing protein nonspecific adsorption.

  9. Selection of site specific vibration equation by using analytic hierarchy process in a quarry

    Energy Technology Data Exchange (ETDEWEB)

    Kalayci, Ulku, E-mail: ukalayci@istanbul.edu.tr; Ozer, Umit, E-mail: uozer@istanbul.edu.tr

    2016-01-15

    This paper presents a new approach for the selection of the most accurate SSVA (Site Specific Vibration Attenuation) equation for blasting processes in a quarry located near settlements in Istanbul, Turkey. In this context, the SSVA equations obtained from the same study area in the literature were considered in terms of distance between the shot points and buildings and the amount of explosive charge. In this purpose, 11 different SSVA equations obtained from the study area in the past 12 years, forecasting capabilities according to designated new conditions, using 102 vibration records as test data obtained from the study area was investigated. In this study, AHP (Analytic Hierarchy Process) was selected as an analysis method in order to determine the most accurate equation among 11 SSAV equations, and the parameters such as year, distance, charge, and r{sup 2} of the equations were used as criteria for AHP. Finally, the most appropriate equation was selected among the existing ones, and the process of selecting according to different target criteria was presented. Furthermore, it was noted that the forecasting results of the selected equation is more accurate than that formed using the test results. - Highlights: • The optimum Site Specific Vibration Attenuation equation for blasting in a quarry located near settlements was determined. • It is indicated that SSVA equations changing over the years don’t give always accurate estimates at changing conditions. • Selection of the blast induced SSVA equation was made using AHP. • Equation selection method was highlighted based on parameters such as charge, distance, and quarry geometry changes (year).

  10. Selection of site specific vibration equation by using analytic hierarchy process in a quarry

    International Nuclear Information System (INIS)

    Kalayci, Ulku; Ozer, Umit

    2016-01-01

    This paper presents a new approach for the selection of the most accurate SSVA (Site Specific Vibration Attenuation) equation for blasting processes in a quarry located near settlements in Istanbul, Turkey. In this context, the SSVA equations obtained from the same study area in the literature were considered in terms of distance between the shot points and buildings and the amount of explosive charge. In this purpose, 11 different SSVA equations obtained from the study area in the past 12 years, forecasting capabilities according to designated new conditions, using 102 vibration records as test data obtained from the study area was investigated. In this study, AHP (Analytic Hierarchy Process) was selected as an analysis method in order to determine the most accurate equation among 11 SSAV equations, and the parameters such as year, distance, charge, and r"2 of the equations were used as criteria for AHP. Finally, the most appropriate equation was selected among the existing ones, and the process of selecting according to different target criteria was presented. Furthermore, it was noted that the forecasting results of the selected equation is more accurate than that formed using the test results. - Highlights: • The optimum Site Specific Vibration Attenuation equation for blasting in a quarry located near settlements was determined. • It is indicated that SSVA equations changing over the years don’t give always accurate estimates at changing conditions. • Selection of the blast induced SSVA equation was made using AHP. • Equation selection method was highlighted based on parameters such as charge, distance, and quarry geometry changes (year).

  11. 29 CFR 1926.752 - Site layout, site-specific erection plan and construction sequence.

    Science.gov (United States)

    2010-07-01

    ... 29 Labor 8 2010-07-01 2010-07-01 false Site layout, site-specific erection plan and construction... Steel Erection § 1926.752 Site layout, site-specific erection plan and construction sequence. (a... strength or sufficient strength to support the loads imposed during steel erection. (c) Site layout. The...

  12. 76 FR 5147 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2011-01-28

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Paducah AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act... recommendations to DOE-EM and site management in the areas of environmental restoration, waste management and...

  13. 77 FR 59598 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2012-09-28

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Paducah AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act... recommendations to DOE-EM and site management in the areas of environmental restoration, waste management and...

  14. 75 FR 13269 - Environmental Management Site-Specific Advisory Board, Hanford

    Science.gov (United States)

    2010-03-19

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Hanford AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Hanford. The Federal Advisory Committee Act... is to make recommendations to DOE-EM and site management in the areas of environmental restoration...

  15. 75 FR 65310 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2010-10-22

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Nevada AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Nevada Test Site. The Federal Advisory... Board is to make recommendations to DOE-EM and site management in the areas of environmental restoration...

  16. 75 FR 54600 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2010-09-08

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Paducah AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act... recommendations to DOE-EM and site management in the areas of environmental restoration, waste management and...

  17. 75 FR 66074 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2010-10-27

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Paducah AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act... recommendations to DOE-EM and site management in the areas of environmental restoration, waste management and...

  18. 75 FR 8050 - Environmental Management Site-Specific Advisory Board, Hanford

    Science.gov (United States)

    2010-02-23

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Hanford AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Hanford. The Federal Advisory Committee Act... is to make recommendations to DOE-EM and site management in the areas of environmental restoration...

  19. 75 FR 24686 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2010-05-05

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Paducah AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act... recommendations to DOE-EM and site management in the areas of environmental restoration, waste management and...

  20. 76 FR 80355 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2011-12-23

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Paducah AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act... make recommendations to DOE-EM and site management in the areas of environmental restoration, waste...

  1. 75 FR 9404 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2010-03-02

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Paducah AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act... recommendations to DOE-EM and site management in the areas of environmental restoration, waste management and...

  2. 75 FR 56526 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2010-09-16

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Portsmouth AGENCY... Initiative Workshop of the Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The... recommendations to DOE-EM and site management in the areas of environmental restoration, waste management and...

  3. 75 FR 82004 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2010-12-29

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Nevada AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Nevada. The Federal Advisory Committee Act (Pub... recommendations to DOE-EM and site management in the areas of environmental restoration, waste management, and...

  4. 77 FR 4027 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2012-01-26

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Nevada AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Nevada. The Federal Advisory Committee Act (Pub... recommendations to DOE-EM and site management in the areas of environmental restoration, waste management, and...

  5. 77 FR 43583 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2012-07-25

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Paducah AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act... recommendations to DOE-EM and site management in the areas of environmental restoration, waste management and...

  6. 75 FR 61711 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2010-10-06

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Paducah AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act... recommendations to DOE-EM and site management in the areas of environmental restoration, waste management and...

  7. 76 FR 80354 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2011-12-23

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Nevada AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Nevada. The Federal Advisory Committee Act (Pub... recommendations to DOE-EM and site management in the areas of environmental restoration, waste management, and...

  8. 75 FR 82002 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2010-12-29

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Paducah AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act... recommendations to DOE-EM and site management in the areas of environmental restoration, waste management and...

  9. 76 FR 61350 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2011-10-04

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Paducah AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act... make recommendations to DOE-EM and site management in the areas of environmental restoration, waste...

  10. 76 FR 4645 - Environmental Management Site-Specific Advisory Board, Hanford

    Science.gov (United States)

    2011-01-26

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Hanford AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Hanford. The Federal Advisory Committee Act... recommendations to DOE-EM and site management in the areas of environmental restoration, waste management, and...

  11. 77 FR 2282 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2012-01-17

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Paducah AGENCY... the Environmental Management Site-Specific Advisory Board, Paducah. This notice announces the... Management Officer. [FR Doc. 2012-831 Filed 1-12-12; 4:15 pm] BILLING CODE 6405-01-P ...

  12. 76 FR 20651 - Environmental Management Site-Specific Advisory Board Chairs

    Science.gov (United States)

    2011-04-13

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board Chairs AGENCY... a meeting on April 13-14, 2011 of the Environmental Management Site-Specific Advisory Board Chairs... R. Butler, Acting Deputy Committee Management Officer. [FR Doc. 2011-8970 Filed 4-8-11; 4:15 pm...

  13. 75 FR 6018 - Environmental Management Site-Specific Advisory Board, Hanford

    Science.gov (United States)

    2010-02-05

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Hanford AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Hanford (known locally as the Hanford Advisory... and site management in the areas of environmental restoration, waste management, and related...

  14. 77 FR 12044 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2012-02-28

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Nevada AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Nevada. The Federal Advisory Committee Act (Pub... Board is to make recommendations to DOE-EM and site management in the areas of environmental restoration...

  15. 76 FR 48148 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2011-08-08

    ... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Paducah AGENCY... Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act... recommendations to DOE-EM and site management in the areas of environmental restoration, waste management and...

  16. FLP recombinase-mediated site-specific recombination in silkworm, Bombyx mori

    Science.gov (United States)

    A comprehensive understanding of gene function and the production of site-specific genetically modified mutants are two major goals of genetic engineering in the post-genomic era. Although site-specific recombination systems have been powerful tools for genome manipulation of many organisms, they h...

  17. New labeling and separation methods for in vivo and in vitro diagnostics in Hungary

    International Nuclear Information System (INIS)

    Veres, A.; Toth, G.; Zsinka, L.; Miller, J.

    1986-01-01

    Three methods have been developed: 1. An adsorption chromatographic method for the separation of iodine-125-labeled compounds applied as tracers in the radioimmunoassay; 2. A portable sublimation generator for the separation of technetium-99m from low or medium specific activity molybdenum-99 using titanium molybdate as a new target material; 3. A novel dry distillation method for the production of iodine-131 from melted, pile-irradiated TeO 2 . The method renders possible to get rid of liquid radioactive wastes. 1 reference

  18. Saturation recovery EPR spin-labeling method for quantification of lipids in biological membrane domains.

    Science.gov (United States)

    Mainali, Laxman; Camenisch, Theodore G; Hyde, James S; Subczynski, Witold K

    2017-12-01

    The presence of integral membrane proteins induces the formation of distinct domains in the lipid bilayer portion of biological membranes. Qualitative application of both continuous wave (CW) and saturation recovery (SR) electron paramagnetic resonance (EPR) spin-labeling methods allowed discrimination of the bulk, boundary, and trapped lipid domains. A recently developed method, which is based on the CW EPR spectra of phospholipid (PL) and cholesterol (Chol) analog spin labels, allows evaluation of the relative amount of PLs (% of total PLs) in the boundary plus trapped lipid domain and the relative amount of Chol (% of total Chol) in the trapped lipid domain [ M. Raguz, L. Mainali, W. J. O'Brien, and W. K. Subczynski (2015), Exp. Eye Res., 140:179-186 ]. Here, a new method is presented that, based on SR EPR spin-labeling, allows quantitative evaluation of the relative amounts of PLs and Chol in the trapped lipid domain of intact membranes. This new method complements the existing one, allowing acquisition of more detailed information about the distribution of lipids between domains in intact membranes. The methodological transition of the SR EPR spin-labeling approach from qualitative to quantitative is demonstrated. The abilities of this method are illustrated for intact cortical and nuclear fiber cell plasma membranes from porcine eye lenses. Statistical analysis (Student's t -test) of the data allowed determination of the separations of mean values above which differences can be treated as statistically significant ( P ≤ 0.05) and can be attributed to sources other than preparation/technique.

  19. Design of site specific radiopharmaceuticals for tumor imaging. (Parts I and II)

    International Nuclear Information System (INIS)

    Van Dort, M.E.

    1983-01-01

    Part I. Synthetic methods were developed for the preparation of several iodinated benzoic acid hydrazides as labeling moieties for indirect tagging of carbonyl-containing bio-molecules and potential tumor-imaging agents. Biodistribution studies conducted in mice on the derivatives having the I-125 label ortho to a phenolic OH demonstrated a rapid in vivo deiodination. Part II. The reported high melanin binding affinity of quinoline and other heterocyclic antimalarial drugs led to the development of many analogues of such molecules as potential melanoma-imaging agents. Once such analogue iodochloroquine does exhibit high melanin binding, but has found limited clinical use due to appreciable accumulation in non-target tissues such as the adrenal cortex and inner ear. This project developed a new series of candidate melanoma imaging agents which would be easier to radio-label, could yield higher specific activity product, and which might demonstrate more favorable pharmacokinetic and dosimetric characteristics compared to iodochloroquine

  20. Site-Specific Difference of Bone Geometry Indices in Hypoparathyroid Patients

    Directory of Open Access Journals (Sweden)

    Hye-Sun Park

    2017-02-01

    Full Text Available BackgroundHypoparathyroid patients often have a higher bone mineral density (BMD than the general population. However, an increase in BMD does not necessarily correlate with a solid bone microstructure. This study aimed to evaluate the bone microstructure of hypoparathyroid patients by using hip structure analysis (HSA.MethodsNinety-five hypoparathyroid patients >20 years old were enrolled and 31 of them had eligible data for analyzing bone geometry parameters using HSA. And among the control data, we extracted sex-, age-, and body mass index-matched three control subjects to each patient. The BMD data were reviewed retrospectively and the bone geometry parameters of the patients were analyzed by HSA.ResultsThe mean Z-scores of hypoparathyroid patients at the lumbar spine, femoral neck, and total hip were above zero (0.63±1.17, 0.48±1.13, and 0.62±1.10, respectively. The differences in bone geometric parameters were site specific. At the femoral neck and intertrochanter, the cross-sectional area (CSA and cortical thickness (C.th were higher, whereas the buckling ratio (BR was lower than in controls. However, those trends were opposite at the femoral shaft; that is, the CSA and C.th were low and the BR was high.ConclusionOur study shows the site-specific effects of hypoparathyroidism on the bone. Differences in bone components, marrow composition, or modeling based bone formation may explain these findings. However, further studies are warranted to investigate the mechanism, and its relation to fracture risk.

  1. Characterization and validation of new tools for measuring site-specific cardiac troponin I phosphorylation.

    Science.gov (United States)

    Thoemmes, Stephen F; Stutzke, Crystal A; Du, Yanmei; Browning, Michael D; Buttrick, Peter M; Walker, Lori A

    2014-01-31

    Phosphorylation of cardiac troponin I is a well established mechanism by which cardiac contractility is modulated. However, there are a number of phosphorylation sites on TnI which contribute singly or in combination to influence cardiac function. Accordingly, methods for accurately measuring site-specific TnI phosphorylation are needed. Currently, two strategies are employed: mass spectrometry, which is costly, difficult and has a low throughput; and Western blotting using phospho-specific antibodies, which is limited by the availability of reagents. In this report, we describe a cohort of new site-specific TnI phosphoantibodies, generated against physiologically relevant phosphorylation sites, that are superior to the current commercially available antibodies: to phospho-serine 22/23 which shows a >5-fold phospho-specificity for phosphorylated TnI; to phospho-serine 43, which has >3-fold phospho-specificity for phosphorylated TnI; and phospho-serine 150 which has >2-fold phospho-specificity for phosphorylated TnI. These new antibodies demonstrated greater sensitivity and specificity for the phosphorylated TnI than the most widely used commercially available reagents. For example, at a protein load of 20 μg of total cardiac extract, a commercially available antibody recognized both phosphorylated and dephosphorylated TnI to the same degree. At the same protein load our phospho-serine 22/23 antibody exhibited no cross-reactivity with dephosphorylated TnI. These new tools should allow a more accurate assessment and a better understanding of the role of TnI phosphorylation in the response of the heart to pathologic stress. Copyright © 2013 Elsevier B.V. All rights reserved.

  2. A simple method for labelling proteins with 211At via diazotized aromatic diamine

    International Nuclear Information System (INIS)

    Wunderlich, G.; Franke, W.-G.; Fischer, S.; Dreyer, R.

    1987-01-01

    A simple and rapid method for labelling proteins with 211 At by means of a 1,4-diaminobenzene link is described. This link is transformed into the diazonium salt and subsequently reactions of both 211 At and proteins with the diazonium salt take place simultaneously. For possibly high yields of astatized protein an appropriate temperature of 273 K was found. The results demonstrate the difference between the reaction mechanisms of iodine and astatine with proteins. (author)

  3. Energy expenditure and socioeconomic status in Guatemala as measured by the doubly labelled water method

    International Nuclear Information System (INIS)

    Stein, T.P.; Johnston, F.E.; Greiner, L.

    1988-01-01

    The energy expenditure of lower (group 1) and upper socioeconomic group females (group 2) from a marginal community in Guatemala City was determined by using the doubly labelled water method. Energy expenditure values were 1925 +/- 66 (mean, SEM) kcal/d (group 1) and 2253 +/- 145 kcal/d group 2 (p less than 0.03). About half of this difference can be attributed to size

  4. Tumor scintigraphy by the method for subtracting the initial image with technetium-99m labeled antibody

    International Nuclear Information System (INIS)

    Karube, Yoshiharu; Katsuno, Kentaro; Ito, Sanae; Matsunaga, Kazuhisa; Takata, Jiro; Kuroki, Masahide; Murakami, Masaaki; Matsuoka, Yuji

    1999-01-01

    The method for subtracting the initial image from the localization image was evaluated for radioimmunoscintigraphy of tumors with technetium-99m (Tc-99m) labeled antibodies. Monoclonal antibodies were parental mouse and mouse-human chimeric antibodies to carcinoembryonic antigen (CEA), designated F11-39 and ChF11-39, respectively, both of which have been found to discriminate CEA in tumor tissues from the CEA-related antigens. After reduction of the intrinsic disulfide bonds, these antibodies were labeled with Tc-99m. In vivo studies were performed on athymic nude mice bearing the human CEA-producing gastric carcinoma xenografts. Though biodistribution results showed selective and progressive accumulation of Tc-99m labeled antibodies at the tumor site, high radioactivity in blood was inappropriate for scintigraphic visualization of the tumors within a few hours. We examined the subtraction of the initial Tc-99m image from the Tc-99m localization image after a few hours. Subtracted images of the same count reflected the in vivo behavior of the Tc-99m radioactivity. The subtracted scintigrams revealed excellent tumor images with no significant extrarenal background. Visualization of the tumor site was dependent on antigen-specific binding and nonspecific exudation. These results demonstrate that a method of subtraction of the initial image may serve as a potentially useful diagnostic method for an abnormal site for agents with a low pharmacokinetic value. (author)

  5. Multimodal nonlinear microscopy: A powerful label-free method for supporting standard diagnostics on biological tissues

    Directory of Open Access Journals (Sweden)

    Riccardo Cicchi

    2014-09-01

    Full Text Available The large use of nonlinear laser scanning microscopy in the past decade paved the way for potential clinical application of this imaging technique. Modern nonlinear microscopy techniques offer promising label-free solutions to improve diagnostic performances on tissues. In particular, the combination of multiple nonlinear imaging techniques in the same microscope allows integrating morphological with functional information in a morpho-functional scheme. Such approach provides a high-resolution label-free alternative to both histological and immunohistochemical examination of tissues and is becoming increasingly popular among the clinical community. Nevertheless, several technical improvements, including automatic scanning and image analysis, are required before the technique represents a standard diagnostic method. In this review paper, we highlight the capabilities of multimodal nonlinear microscopy for tissue imaging, by providing various examples on colon, arterial and skin tissues. The comparison between images acquired using multimodal nonlinear microscopy and histology shows a good agreement between the two methods. The results demonstrate that multimodal nonlinear microscopy is a powerful label-free alternative to standard histopathological methods and has the potential to find a stable place in the clinical setting in the near future.

  6. Validity of the Remote Food Photography Method against Doubly Labeled Water among Minority Preschoolers

    OpenAIRE

    Nicklas, Theresa; Saab, Rabab; Islam, Noemi G.; Wong, William; Butte, Nancy; Schulin, Rebecca; Liu, Yan; Apolzan, John W.; Myers, Candice A.; Martin, Corby K.

    2017-01-01

    Objective To determine the validity of energy intake (EI) estimations made using the Remote Food Photography Method (RFPM) compared to the doubly-labeled water (DLW) method in minority preschool children in a free-living environment. Methods Seven days of food intake and spot urine samples excluding first void collections for DLW analysis were obtained on 39 3-to-5 year old Hispanic and African American children. Using an iPhone, caregivers captured before and after pictures of the child’s in...

  7. A flexible fluorescence correlation spectroscopy based method for quantification of the DNA double labeling efficiency with precision control

    International Nuclear Information System (INIS)

    Hou, Sen; Tabaka, Marcin; Sun, Lili; Trochimczyk, Piotr; Kaminski, Tomasz S; Kalwarczyk, Tomasz; Zhang, Xuzhu; Holyst, Robert

    2014-01-01

    We developed a laser-based method to quantify the double labeling efficiency of double-stranded DNA (dsDNA) in a fluorescent dsDNA pool with fluorescence correlation spectroscopy (FCS). Though, for quantitative biochemistry, accurate measurement of this parameter is of critical importance, before our work it was almost impossible to quantify what percentage of DNA is doubly labeled with the same dye. The dsDNA is produced by annealing complementary single-stranded DNA (ssDNA) labeled with the same dye at 5′ end. Due to imperfect ssDNA labeling, the resulting dsDNA is a mixture of doubly labeled dsDNA, singly labeled dsDNA and unlabeled dsDNA. Our method allows the percentage of doubly labeled dsDNA in the total fluorescent dsDNA pool to be measured. In this method, we excite the imperfectly labeled dsDNA sample in a focal volume of <1 fL with a laser beam and correlate the fluctuations of the fluorescence signal to get the FCS autocorrelation curves; we express the amplitudes of the autocorrelation function as a function of the DNA labeling efficiency; we perform a comparative analysis of a dsDNA sample and a reference dsDNA sample, which is prepared by increasing the total dsDNA concentration c (c > 1) times by adding unlabeled ssDNA during the annealing process. The method is flexible in that it allows for the selection of the reference sample and the c value can be adjusted as needed for a specific study. We express the precision of the method as a function of the ssDNA labeling efficiency or the dsDNA double labeling efficiency. The measurement precision can be controlled by changing the c value. (letter)

  8. FLP recombinase-mediated site-specific recombination in silkworm, Bombyx mori.

    Directory of Open Access Journals (Sweden)

    Ding-Pei Long

    Full Text Available A comprehensive understanding of gene function and the production of site-specific genetically modified mutants are two major goals of genetic engineering in the post-genomic era. Although site-specific recombination systems have been powerful tools for genome manipulation of many organisms, they have not yet been established for use in the manipulation of the silkworm Bombyx mori genome. In this study, we achieved site-specific excision of a target gene at predefined chromosomal sites in the silkworm using a FLP/FRT site-specific recombination system. We first constructed two stable transgenic target silkworm strains that both contain a single copy of the transgene construct comprising a target gene expression cassette flanked by FRT sites. Using pre-blastoderm microinjection of a FLP recombinase helper expression vector, 32 G3 site-specific recombinant transgenic individuals were isolated from five of 143 broods. The average frequency of FLP recombinase-mediated site-specific excision in the two target strains genome was approximately 3.5%. This study shows that it is feasible to achieve site-specific recombination in silkworms using the FLP/FRT system. We conclude that the FLP/FRT system is a useful tool for genome manipulation in the silkworm. Furthermore, this is the first reported use of the FLP/FRT system for the genetic manipulation of a lepidopteran genome and thus provides a useful reference for the establishment of genome manipulation technologies in other lepidopteran species.

  9. Studies on the clinical application of MR perfusion image using arterial spin labeling method

    International Nuclear Information System (INIS)

    Miyasaka, Kenji

    1999-01-01

    A new technique for imaging brain perfusion, arterial spin labeling method was applied in clinic. Brain perfusion was imaged by FAIR and EPISTAR both of which using arterial spin labeling (ASL) method. Suitable parameters for small contamination were examined using a imaging phantom. Then normal volunteers were examined for imaging timing. Suitable time between labeling pulse and imaging pulse for brain capillary and parenchyma was 1.0 sec. For clinical application study, total 48 patients with brain diseases were examined by FAIR and/or EPISTAR. A lesion/white matter signal intensity ratio was calculated in all clinical cases. Average of signal intensity ratio in infarction, tumor and arteriovenous malformation (AVM) were 0.8, 2.2 and 18.6 at FAIR, and 0.6, 2.2 and 12.8 at EPISTAR, respectively. Low perfusion diseases such as cerebral infarction have low signal intensity ratio and high perfusion diseases such as AVM have high signal intensity ratio in both FAIR and EPISTAR. Brain lesions were imaged similarly in FAIR and EPISTAR, and no remarkable difference was found between FAIR and EPISTAR. As a result of diagnostic trial by signal intensity ratio in operated tumor, hemorrhagic cases could be diagnosed by accuracies of 75% in FAIR and 100% in EPISTAR, respectively. (author)

  10. One-carbon 13C-labeled synthetic intermediates. Comparison and evaluation of preparative methods

    International Nuclear Information System (INIS)

    Ott, D.G.

    1978-01-01

    Frequently the biggest stumbling block to the synthesis of a structurally complex labeled compound is obtaining the required low molecular weight, structurally simple, isotopic intermediates. Selection of a particular scheme from various alternatives depends on the available capabilities and quantity of product desired, as well as on anticipated future requirements and need for related compounds. Many of the newer reagents for organic synthesis can be applied effectively to isotopic preparations with improvements of yields and simplification of procedures compared to established classical methods. New routes developed for higher molecular weight compounds are sometimes not directly adaptable to the one-carbon analogs, either because of isolation difficulties occasioned by physical properties or by chemical reactivities peculiar to their being first members of homologous series. Various routes for preparation of carbon-13 labeled methanol, formaldehyde, and cyanide are compared

  11. Radioactive indium labelling of the figured elements of blood. Method, results, applications

    International Nuclear Information System (INIS)

    Ducassou, D.; Nouel, J.P.

    Following the work of Thakur et al. the authors became interested in red corpuscle, leucocyte and platelet labelling with indium 111 or 113m (8 hydroxyquinolein-indium). For easier labelling of the figured elements of blood the technique described was modified. The chelate is prepared by simple contact at room temperature of indium 111 or 113m chloride and water-soluble 8 hydroxyquinolein sulphate, in the presence of 0.2M TRIS buffer. The figured element chosen suspended in physiological serum is added directly to the solution obtained, the platelets and leucocytes being separated out beforehand by differential centrifugation. While it gives results similar to those of Thabur et al. the method proposed avoids the chloroform extraction of the radioactive chelate and the use of alcohol, liable to impair the platelet regation capacity [fr

  12. Optimization of photoactive protein Z for fast and efficient site-specific conjugation of native IgG.

    Science.gov (United States)

    Hui, James Z; Tsourkas, Andrew

    2014-09-17

    Antibody conjugates have been used in a variety of applications from immunoassays to drug conjugates. However, it is becoming increasingly clear that in order to maximize an antibody's antigen binding ability and to produce homogeneous antibody-conjugates, the conjugated molecule should be attached onto IgG site-specifically. We previously developed a facile method for the site-specific modification of full length, native IgGs by engineering a recombinant Protein Z that forms a covalent link to the Fc domain of IgG upon exposure to long wavelength UV light. To further improve the efficiency of Protein Z production and IgG conjugation, we constructed a panel of 13 different Protein Z variants with the UV-active amino acid benzoylphenylalanine (BPA) in different locations. By using this panel of Protein Z to cross-link a range of IgGs from different hosts, including human, mouse, and rat, we discovered two previously unknown Protein Z variants, L17BPA and K35BPA, that are capable of cross-linking many commonly used IgG isotypes with efficiencies ranging from 60% to 95% after only 1 h of UV exposure. When compared to existing site-specific methods, which often require cloning or enzymatic reactions, the Protein Z-based method described here, utilizing the L17BPA, K35BPA, and the previously described Q32BPA variants, represents a vastly more accessible and efficient approach that is compatible with nearly all native IgGs, thus making site-specific conjugation more accessible to the general research community.

  13. Site-specific RNase A activity was dramatically reduced in serum from multiple types of cancer patients.

    Directory of Open Access Journals (Sweden)

    Weiyan Huang

    Full Text Available Potent RNase activities were found in the serum of mammals but the physiological function of the RNases was never well illustrated, largely due to the caveats in methods of RNase activity measurement. None of the existing methods can distinguish between RNases with different target specificities. A systematic study was recently carried out in our lab to investigate the site-specificity of serum RNases on double-stranded RNA substrates, and found that serum RNases cleave double-stranded RNAs predominantly at 5'-U/A-3' and 5'-C/A-3' dinucleotide sites, in a manner closely resembling RNase A. Based on this finding, a FRET assay was developed in the current study to measure this site-specific serum RNase activity in human samples using a double stranded RNA substrate. We demonstrated that the method has a dynamic range of 10(-5 mg/ml- 10(-1 mg/ml using serial dilution of RNase A. The sera of 303 cancer patients were subjected to comparison with 128 healthy controls, and it was found that serum RNase activities visualized with this site-specific double stranded probe were found to be significantly reduced in patients with gastric cancer, liver cancer, pancreatic cancer, esophageal cancer, ovary cancer, cervical cancer, bladder cancer, kidney cancer and lung cancer, while only minor changes were found in breast and colon cancer patients. This is the first report using double stranded RNA as probe to quantify site-specific activities of RNase A in a serum. The results illustrated that RNase A might be further evaluated to determine if it can serve as a new class of biomarkers for certain cancer types.

  14. Site-Specific RNase A Activity Was Dramatically Reduced in Serum from Multiple Types of Cancer Patients

    Science.gov (United States)

    Huang, Weiyan; Zhao, Mei; Wei, Na; Wang, Xiaoxia; Cao, Huqing; Du, Quan; Liang, Zicai

    2014-01-01

    Potent RNase activities were found in the serum of mammals but the physiological function of the RNases was never well illustrated, largely due to the caveats in methods of RNase activity measurement. None of the existing methods can distinguish between RNases with different target specificities. A systematic study was recently carried out in our lab to investigate the site-specificity of serum RNases on double-stranded RNA substrates, and found that serum RNases cleave double-stranded RNAs predominantly at 5′-U/A-3′ and 5′-C/A-3′ dinucleotide sites, in a manner closely resembling RNase A. Based on this finding, a FRET assay was developed in the current study to measure this site-specific serum RNase activity in human samples using a double stranded RNA substrate. We demonstrated that the method has a dynamic range of 10−5 mg/ml- 10−1 mg/ml using serial dilution of RNase A. The sera of 303 cancer patients were subjected to comparison with 128 healthy controls, and it was found that serum RNase activities visualized with this site-specific double stranded probe were found to be significantly reduced in patients with gastric cancer, liver cancer, pancreatic cancer, esophageal cancer, ovary cancer, cervical cancer, bladder cancer, kidney cancer and lung cancer, while only minor changes were found in breast and colon cancer patients. This is the first report using double stranded RNA as probe to quantify site-specific activities of RNase A in a serum. The results illustrated that RNase A might be further evaluated to determine if it can serve as a new class of biomarkers for certain cancer types. PMID:24805924

  15. Nucleophilic Fluorination Reactions in Novel Reaction Media for 18F-Fluorine Labeling Method

    International Nuclear Information System (INIS)

    Kim, Dong Wook; Jeong, Hwan Jeong; Lim, Seok Tae; Sohn, Myung Hee

    2009-01-01

    Noninvasive imaging of molecular and biological processes in living subjects with positron emission tomography (PET) provides exciting opportunities to monitor metabolism and detect diseases in humans. Measuring these processes with PET requires the preparation of specific molecular imaging probes labeled with 18F-fluorine. In this review we describe recent methods and novel trends for the introduction of 18 F-fluorine into molecules which in turn are intended to serve as imaging agents for PET study. Nucleophilic 18 F-fluorination of some halo- and mesyloxyalkanes to the corresponding 18 F-fluoroalkanes with 18 F-fluoride obtained from an 18 O(p,n) 18 F reaction, using novel reaction media system such as an ionic liquidor tert-alcohol, has been studied as a new method for 18 F-fluorine labeling. Ionic liquid method is rapid and particularly convenient because 18 F-fluoride in H 2 O can be added directly to the reaction media, obviating the careful drying that is typically required for currently used radiofluorination methods. The nonpolar protic tert-alcohol enhances the nucleophilicity of the fluoride ion dramatically in the absence of any kind of catalyst, greatly increasing the rate of the nucleophilic fluorination and reducing formation of byproducts compared with conventional methods using dipolar aprotic solvents. The great efficacy of this method is a particular advantage in labeling radiopharmaceuticals with 18 F-fluorine for PET imaging, and it is illustrated by the synthesis of 18 F-fluoride radiolabeled molecular imaging probes, such as 18 F-FDG, 18 F-FLT, 18 F-FP-CIT, and 18 F-FMISO, in high yield and purity and in shorter times compared to conventional syntheses

  16. A multi-label learning based kernel automatic recommendation method for support vector machine.

    Science.gov (United States)

    Zhang, Xueying; Song, Qinbao

    2015-01-01

    Choosing an appropriate kernel is very important and critical when classifying a new problem with Support Vector Machine. So far, more attention has been paid on constructing new kernels and choosing suitable parameter values for a specific kernel function, but less on kernel selection. Furthermore, most of current kernel selection methods focus on seeking a best kernel with the highest classification accuracy via cross-validation, they are time consuming and ignore the differences among the number of support vectors and the CPU time of SVM with different kernels. Considering the tradeoff between classification success ratio and CPU time, there may be multiple kernel functions performing equally well on the same classification problem. Aiming to automatically select those appropriate kernel functions for a given data set, we propose a multi-label learning based kernel recommendation method built on the data characteristics. For each data set, the meta-knowledge data base is first created by extracting the feature vector of data characteristics and identifying the corresponding applicable kernel set. Then the kernel recommendation model is constructed on the generated meta-knowledge data base with the multi-label classification method. Finally, the appropriate kernel functions are recommended to a new data set by the recommendation model according to the characteristics of the new data set. Extensive experiments over 132 UCI benchmark data sets, with five different types of data set characteristics, eleven typical kernels (Linear, Polynomial, Radial Basis Function, Sigmoidal function, Laplace, Multiquadric, Rational Quadratic, Spherical, Spline, Wave and Circular), and five multi-label classification methods demonstrate that, compared with the existing kernel selection methods and the most widely used RBF kernel function, SVM with the kernel function recommended by our proposed method achieved the highest classification performance.

  17. Site-specific tagging proteins with a rigid, small and stable transition metal chelator, 8-hydroxyquinoline, for paramagnetic NMR analysis

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Yin; Huang, Feng [Nankai University, State Key Laboratory of Elemento-Organic Chemistry, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) (China); Huber, Thomas [Australian National University, Research School of Chemistry (Australia); Su, Xun-Cheng, E-mail: xunchengsu@nankai.edu.cn [Nankai University, State Key Laboratory of Elemento-Organic Chemistry, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) (China)

    2016-02-15

    Design of a paramagnetic metal binding motif in a protein is a valuable way for understanding the function, dynamics and interactions of a protein by paramagnetic NMR spectroscopy. Several strategies have been proposed to site-specifically tag proteins with paramagnetic lanthanide ions. Here we report a simple approach of engineering a transition metal binding motif via site-specific labelling of a protein with 2-vinyl-8-hydroxyquinoline (2V-8HQ). The protein-2V-8HQ adduct forms a stable complex with transition metal ions, Mn(II), Co(II), Ni(II), Cu(II) and Zn(II). The paramagnetic effects generated by these transition metal ions were evaluated by NMR spectroscopy. We show that 2V-8HQ is a rigid and stable transition metal binding tag. The coordination of the metal ion can be assisted by protein sidechains. More importantly, tunable paramagnetic tensors are simply obtained in an α-helix that possesses solvent exposed residues in positions i and i + 3, where i is the residue to be mutated to cysteine, i + 3 is Gln or Glu or i − 4 is His. The coordination of a sidechain carboxylate/amide or imidazole to cobalt(II) results in different structural geometries, leading to different paramagnetic tensors as shown by experimental data.

  18. Quantitative chemoproteomics for site-specific analysis of protein alkylation by 4-hydroxy-2-nonenal in cells.

    Science.gov (United States)

    Yang, Jing; Tallman, Keri A; Porter, Ned A; Liebler, Daniel C

    2015-03-03

    Protein alkylation by 4-hydroxy-2-nonenal (HNE), an endogenous lipid derived electrophile, contributes to stress signaling and cellular toxicity. Although previous work has identified protein targets for HNE alkylation, the sequence specificity of alkylation and dynamics in a cellular context remain largely unexplored. We developed a new quantitative chemoproteomic platform, which uses isotopically tagged, photocleavable azido-biotin reagents to selectively capture and quantify the cellular targets labeled by the alkynyl analogue of HNE (aHNE). Our analyses site-specifically identified and quantified 398 aHNE protein alkylation events (386 cysteine sites and 12 histidine sites) in intact cells. This data set expands by at least an order of magnitude the number of such modification sites previously reported. Although adducts formed by Michael addition are thought to be largely irreversible, we found that most aHNE modifications are lost rapidly in situ. Moreover, aHNE adduct turnover occurs only in intact cells and loss rates are site-selective. This quantitative chemoproteomics platform provides a versatile general approach to map bioorthogonal-chemically engineered post-translational modifications and their cellular dynamics in a site-specific and unbiased manner.

  19. N-[3H]acetyl-labeling, a convenient method for radiolabeling of glycosaminoglycans

    International Nuclear Information System (INIS)

    Hook, M.; Riesenfeld, J.; Lindahl, U.

    1982-01-01

    A method for the introduction of N-[ 3 H]acetyl groups into glycosaminoglycans is described. The procedure is based on [ 3 H]acetylation of N-unsubstituted hexosamine residues by treating the polysaccharides with [ 3 H]acetic anhydride. Preparations of heparin and heparin sulfate were found to contain significant numbers of N-unsubstituted hexosamine residues, as isolates. In contrast, such units could not be detected in chondroitin sulfate, dermatan sulfate, or hyaluronic acid. These polysaccharides were therefore subjected to partial N-deacetylation by reaction with hydrazine in the presence of hydrazine sulfate. After treatment with [ 3 H]acetic anhydride, the specific activities of the resulting labeled polysaccharide preparations ranged between 0.1 X 10 6 and 0.6 X 10 6 cpm 3 H/μg of uronic acid. The 3 H-labeled polysaccharide preparations did not differ significantly from the corresponding unlabeled starting materials with regard to polyanion properties (chromatography on DEAE-cellulose) or polymer chain size (gel chromatography). Further, the radiolabeled polysaccharide derivatives were susceptible to specific enzymatic degradation (chondroitinase ABC and mammalian heparitinase) and retained their ability to interact specifically with certain proteins - for example, [ 3 H]heparin with antithrombin [ 3 H]hyaluronic acid oligosaccharides with chondroitin sulfate proteoglycan. These findings indicate that the labeling procedures did not induce any major structural derangement of the polysaccharide molecules. The method developed should be useful in providing labeled glycosaminoglycans for metabolic and enzymatic experiments as well as for studies on the interacion between glycosaminoglycans and other bilogical macromolecules

  20. Quantitative phosphoproteomics using acetone-based peptide labeling: Method evaluation and application to a cardiac ischemia/reperfusion model

    Science.gov (United States)

    Wijeratne, Aruna B.; Manning, Janet R.; Schultz, Jo El J.; Greis, Kenneth D.

    2013-01-01

    Mass spectrometry (MS) techniques to globally profile protein phosphorylation in cellular systems that are relevant to physiological or pathological changes have been of significant interest in biological research. In this report, an MS-based strategy utilizing an inexpensive acetone-based peptide labeling technique known as reductive alkylation by acetone (RABA) for quantitative phosphoproteomics was explored to evaluate its capacity. Since the chemistry for RABA-labeling for phosphorylation profiling had not been previously reported, it was first validated using a standard phosphoprotein and identical phosphoproteomes from cardiac tissue extracts. A workflow was then utilized to compare cardiac tissue phosphoproteomes from mouse hearts not expressing FGF2 vs. hearts expressing low molecular weight fibroblast growth factor-2 (LMW FGF2) to relate low molecular weight fibroblast growth factor-2 (LMW FGF2) mediated cardioprotective phenomena induced by ischemia/reperfusion (I/R) injury of hearts, with downstream phosphorylation changes in LMW FGF2 signaling cascades. Statistically significant phosphorylation changes were identified at 14 different sites on 10 distinct proteins including some with mechanisms already established for LMW FGF2-mediated cardioprotective signaling (e.g. connexin-43), some with new details linking LMW FGF2 to the cardioprotective mechanisms (e.g. cardiac myosin binding protein C or cMyBPC), and also several new downstream effectors not previously recognized for cardio-protective signaling by LMW FGF2. Additionally, one of the phosphopeptides, cMyBPC/pSer-282, identified was further verified with site-specific quantification using an SRM (selected reaction monitoring)-based approach that also relies on isotope labeling of a synthetic phosphopeptide with deuterated acetone as an internal standard. Overall, this study confirms that the inexpensive acetone-based peptide labeling can be used in both exploratory and targeted quantification

  1. Predicting human splicing branchpoints by combining sequence-derived features and multi-label learning methods.

    Science.gov (United States)

    Zhang, Wen; Zhu, Xiaopeng; Fu, Yu; Tsuji, Junko; Weng, Zhiping

    2017-12-01

    Alternative splicing is the critical process in a single gene coding, which removes introns and joins exons, and splicing branchpoints are indicators for the alternative splicing. Wet experiments have identified a great number of human splicing branchpoints, but many branchpoints are still unknown. In order to guide wet experiments, we develop computational methods to predict human splicing branchpoints. Considering the fact that an intron may have multiple branchpoints, we transform the branchpoint prediction as the multi-label learning problem, and attempt to predict branchpoint sites from intron sequences. First, we investigate a variety of intron sequence-derived features, such as sparse profile, dinucleotide profile, position weight matrix profile, Markov motif profile and polypyrimidine tract profile. Second, we consider several multi-label learning methods: partial least squares regression, canonical correlation analysis and regularized canonical correlation analysis, and use them as the basic classification engines. Third, we propose two ensemble learning schemes which integrate different features and different classifiers to build ensemble learning systems for the branchpoint prediction. One is the genetic algorithm-based weighted average ensemble method; the other is the logistic regression-based ensemble method. In the computational experiments, two ensemble learning methods outperform benchmark branchpoint prediction methods, and can produce high-accuracy results on the benchmark dataset.

  2. Development of a novel DDS for site-specific PEGylated proteins

    Directory of Open Access Journals (Sweden)

    Yoshioka Yasuo

    2011-05-01

    Full Text Available Abstract Because of the shifted focus in life science research from genome analyses to genetic and protein function analyses, we now know functions of numerous proteins. These analyses, including those of newly identified proteins, are expected to contribute to the identification of proteins of therapeutic value in various diseases. Consequently, pharmacoproteomic-based drug discovery and development of protein therapies attracted a great deal of attention in recent years. Clinical applications of most of these proteins are, however, limited because of their unexpectedly low therapeutic effects, resulting from the proteolytic degradation in vivo followed by rapid removal from the circulatory system. Therefore, frequent administration of excessively high dose of a protein is required to observe its therapeutic effect in vivo. This often results in impaired homeostasis in vivo and leads to severe adverse effects. To overcome these problems, we have devised a method for chemical modification of proteins with polyethylene glycol (PEGylation and other water-soluble polymers. In addition, we have established a method for creating functional mutant proteins (muteins with desired properties, and developed a site-specific polymer-conjugation method to further improve their therapeutic potency. In this review, we are introducing our original protein-drug innovation system mentioned above.

  3. Labeling method of 17-allylamino, 17-demethoxygeldanamycin with 131I and its biodistribution in experimental animals

    International Nuclear Information System (INIS)

    Jiang Xinyu; Liu Lu; Gao Wen; Chen Daozhen; Huang Ying; Yang Min; Luo Shineng

    2008-01-01

    Objective: The aims of the study were to find out the optimal 131 I labeling method with 17-allylamino, 17-demethoxygeldanamycin (17-AAG) and also to study its biodistribution in animals. Methods: 131 I-17-AAG was prepared by the reaction of 17-AAG with Na 131 I in the presence of hydrogen peroxide. The labeling efficiency and the stability of 131 I-17-AAG were measured by paper chromatograph. The biodistribution in the ICR normal mice was observed by the blood samplings and major organs that were taken out from mice at 0.5, 1, 4, 8, 24 h after 131 I-17-AAG injection through tail veins. VX2 tumor was also implanted in rabbit liver for in vivo imaging with SPECT. Results: The optimal labeling conditions of 17-AAG with mi were determined. The labeling efficiency was 85.65%. The radiochemical purity of 131 I- 17-AAG in acetoacetate solution was (96.51 ± 0.80)% after purification and its radiochemical purity in normal saline solution was (95.57 ± 0.09)%. The radiochemical purity could keep to 90% in normal saline after 5 d at 4 degree C. The biodistribution study in normal mice showed that the uptake (percentage activity of injection dose per gram of tissue, % ID/g) in liver and kidney was less than that in cholecyst [(3.0963 ± 1.3394) %ID/g] at 0.5 h post-injection, and the uptake in stomach and intestine reached to the highest level at 4 h post-injection. The SPECT images showed that the 131 I-17-AAG was obviously concentrated in the tumor after injection at 2 h and 4 d, 6 d, 14 d with the highest tumor to non-tumor (T/NT) radioactivity ratio of 10.36. Conclusions: The labeling method of 17-AAG with 131 I was successfully established. The 131 I-17-AAG in normal saline had a good stability. The main biodistribution in mice was in digestive system and was excreted through the intestinal tract. The SPECT images showed that 131 I-17-AAG might be a potential target-directed agent to the tumor. (authors)

  4. A site specific model and analysis of the neutral somatic mutation rate in whole-genome cancer data.

    Science.gov (United States)

    Bertl, Johanna; Guo, Qianyun; Juul, Malene; Besenbacher, Søren; Nielsen, Morten Muhlig; Hornshøj, Henrik; Pedersen, Jakob Skou; Hobolth, Asger

    2018-04-19

    Detailed modelling of the neutral mutational process in cancer cells is crucial for identifying driver mutations and understanding the mutational mechanisms that act during cancer development. The neutral mutational process is very complex: whole-genome analyses have revealed that the mutation rate differs between cancer types, between patients and along the genome depending on the genetic and epigenetic context. Therefore, methods that predict the number of different types of mutations in regions or specific genomic elements must consider local genomic explanatory variables. A major drawback of most methods is the need to average the explanatory variables across the entire region or genomic element. This procedure is particularly problematic if the explanatory variable varies dramatically in the element under consideration. To take into account the fine scale of the explanatory variables, we model the probabilities of different types of mutations for each position in the genome by multinomial logistic regression. We analyse 505 cancer genomes from 14 different cancer types and compare the performance in predicting mutation rate for both regional based models and site-specific models. We show that for 1000 randomly selected genomic positions, the site-specific model predicts the mutation rate much better than regional based models. We use a forward selection procedure to identify the most important explanatory variables. The procedure identifies site-specific conservation (phyloP), replication timing, and expression level as the best predictors for the mutation rate. Finally, our model confirms and quantifies certain well-known mutational signatures. We find that our site-specific multinomial regression model outperforms the regional based models. The possibility of including genomic variables on different scales and patient specific variables makes it a versatile framework for studying different mutational mechanisms. Our model can serve as the neutral null model

  5. 76 FR 59392 - Environmental Management Site-Specific Advisory Board, Northern New Mexico

    Science.gov (United States)

    2011-09-26

    ... Welcome and Introductions, Committee Business Items: [cir] Approve October 12, 2011, Meeting Agenda, [cir... DEPARTMENT OF ENERGY Environmental Management Site-Specific Advisory Board, Northern New Mexico... meeting of the Environmental Monitoring, Surveillance and Remediation Committee and Waste Management...

  6. Design and integration of components for site specific control of fertilizer application

    NARCIS (Netherlands)

    Bergeijk, van J.

    2001-01-01

    Keywords: Precision Agriculture, Site Specific Agriculture, Global Positioning System, GPS, Fertilizer Application, Information System.

    Spatial and temporal variability in soil, crop and climate characteristics results in non optimal use of fertilizers when the application

  7. 78 FR 14088 - Environmental Management Site-Specific Advisory Board, Savannah River Site

    Science.gov (United States)

    2013-03-04

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Savannah River Site. The Federal Advisory Committee Act requires that public notice of this meeting be announced in the Federal Register.

  8. Risks of all-cause and site-specific fractures among hospitalized patients with COPD

    OpenAIRE

    Liao, Kuang-Ming; Liang, Fu-Wen; Li, Chung-Yi

    2016-01-01

    Abstract Patients with chronic obstructive pulmonary disease (COPD) have a high prevalence of osteoporosis. The clinical sequel of osteoporosis is fracture. Patients with COPD who experience a fracture also have increased morbidity and mortality. Currently, the types of all-cause and site-specific fracture among patients with COPD are unknown. Thus, we elucidated the all-cause and site-specific fractures among patients with COPD. A retrospective, population-based, cohort study was conducted u...

  9. Site-specific data confirm arsenic exposure predicted by the U.S. Environmental Protection Agency.

    OpenAIRE

    Walker, S; Griffin, S

    1998-01-01

    The EPA uses an exposure assessment model to estimate daily intake to chemicals of potential concern. At the Anaconda Superfund site in Montana, the EPA exposure assessment model was used to predict total and speciated urinary arsenic concentrations. Predicted concentrations were then compared to concentrations measured in children living near the site. When site-specific information on concentrations of arsenic in soil, interior dust, and diet, site-specific ingestion rates, and arsenic abso...

  10. The doubly-labelled water method for measuring energy expenditure. Technical recommendations for use in humans

    International Nuclear Information System (INIS)

    Prentice, A.M.

    1990-01-01

    The doubly-labelled water method using stable isotopes of hydrogen and oxygen, is rapidly becoming established as an important new tool for investigating energy metabolism. It is the first genuinely non-invasive method for measuring energy expenditure in free-living people, providing estimates of habitual expenditure over a time period of 10-20 days. The accuracy and precision of these estimates should be superior to those obtained by traditional factorial methods. The DLW method involves several assumptions about the behaviour of the isotopes, the body water pool and the exchange rates within that pool in the labelled animal. These assumptions are: (1) The volume of the body water pool remains constant throughout the measurement period. (2) The rates of water influx, and water and CO 2 efflux are constant throughout the measurement period. (3) The isotopes label only the H 2 O and CO 2 in the body. (4) The isotopes leave the body only in the form of H 2 O and CO 2 . (5) The concentrations of the isotopes in H 2 O and CO 2 leaving the body are the same as those in body water at that time (i.e. there is no isotopic fractionation). (6) No H 2 O or CO 2 that has left the body re-enters the body. (7) The natural abundance, or ''background'' levels of the isotopes remain constant during the measurement interval. The recommendations presented in this document are based on a variety of criteria including: (a) which procedure among several is theoretically correct in a given application; (b) which procedure is simplest and least prone to methodological errors; and (c) which procedure yields the lowest error in validation studies. Refs, figs and tabs

  11. Microbial profile comparisons of saliva, pooled and site-specific subgingival samples in periodontitis patients.

    Directory of Open Access Journals (Sweden)

    Daniel Belstrøm

    Full Text Available The purpose of this study was to compare microbial profiles of saliva, pooled and site-specific subgingival samples in patients with periodontitis. We tested the hypotheses that saliva can be an alternative to pooled subgingival samples, when screening for presence of periopathogens.Site specific subgingival plaque samples (n = 54, pooled subgingival plaque samples (n = 18 and stimulated saliva samples (n = 18 were collected from 18 patients with generalized chronic periodontitis. Subgingival and salivary microbiotas were characterized by means of HOMINGS (Human Oral Microbe Identification using Next Generation Sequencing and microbial community profiles were compared using Spearman rank correlation coefficient.Pronounced intraindividual differences were recorded in site-specific microbial profiles, and site-specific information was in general not reflected by pooled subgingival samples. Presence of Porphyromonas gingivalis, Treponema denticola, Prevotella intermedia, Filifactor alocis, Tannerella forsythia and Parvimona micra in site-specific subgingival samples were detected in saliva with an AUC of 0.79 (sensitivity: 0.61, specificity: 0.94, compared to an AUC of 0.76 (sensitivity: 0.56, specificity: 0.94 in pooled subgingival samples.Site-specific presence of periodontal pathogens was detected with comparable accuracy in stimulated saliva samples and pooled subgingival plaque samples. Consequently, saliva may be a reasonable surrogate for pooled subgingival samples when screening for presence of periopathogens. Future large-scale studies are needed to confirm findings from this study.

  12. Production, Isolation and Radiolabeling Methods for 211AT- Labeling of Biomolecules

    International Nuclear Information System (INIS)

    Wilbur, D.S.; Hamlin, D.K.; Chyan, M.

    2009-01-01

    Targeted alpha therapy with 211 At-labeled compounds holds great promise for treatment of cancer, particularly compartmentalized cancer (e.g. ovarian cancer), minimal residual cancer after surgery and metastatic disease. Unfortunately, 211 At has limited availability and, due to its unique nature, has the potential to be readily dissociated from the cancer-targeting agents used in vivo. Finding methods to circumvent these two problems has occupied a large amount of our efforts over the past few years. 211 At is produced at the University of Washington on a Scanditronix MC-50 using a 28 MeV alpha beam. Our initial preclinical studies were conducted using a small target assembly with irradiations of a 10 □ A alpha beam, but our desire to ultimately conduct clinical studies led to the design and installation of a new target assembly that had much larger irradiation surface and would withstand beam energies of 50 □A or more. Prior to this upgrade, 211 At was efficiently isolated (60-80%) from the irradiated aluminum-backed bismuth targets by dry distillation at 650 o C. However, the dry distillation method gave low recovery yields (e.g. 10-40%) when the much larger new targets were used. After some attempts to improve the distillation yields, we have more recently conducted a wet chemistry approach to the 211 At isolation. While this method still needs to be optimized, it has provided good recovery (60-90%) of the 211 At. Our radiolabeling methods have undergone a similar transition in the past few years. Until recently our 211 At studies were limited to the use of intact monoclonal antibodies (MAb) labeled using conjugates containing aryl-astatine derivatives due to the deastatination of more rapidly metabolized targeting biomolecules. This limitation made it all but impossible to label important biomolecules such as MAb fragments, engineered proteins, peptides and small molecules. This critical shortcoming of labeling methods for 211 At led to our investigating

  13. Site-specific genomic (SSG and random domain-localized (RDL mutagenesis in yeast

    Directory of Open Access Journals (Sweden)

    Honigberg Saul M

    2004-04-01

    Full Text Available Abstract Background A valuable weapon in the arsenal available to yeast geneticists is the ability to introduce specific mutations into yeast genome. In particular, methods have been developed to introduce deletions into the yeast genome using PCR fragments. These methods are highly efficient because they do not require cloning in plasmids. Results We have modified the existing method for introducing deletions in the yeast (S. cerevisiae genome using PCR fragments in order to target point mutations to this genome. We describe two PCR-based methods for directing point mutations into the yeast genome such that the final product contains no other disruptions. In the first method, site-specific genomic (SSG mutagenesis, a specific point mutation is targeted into the genome. In the second method, random domain-localized (RDL mutagenesis, a mutation is introduced at random within a specific domain of a gene. Both methods require two sequential transformations, the first transformation integrates the URA3 marker into the targeted locus, and the second transformation replaces URA3 with a PCR fragment containing one or a few mutations. This PCR fragment is synthesized using a primer containing a mutation (SSG mutagenesis or is synthesized by error-prone PCR (RDL mutagenesis. In SSG mutagenesis, mutations that are proximal to the URA3 site are incorporated at higher frequencies than distal mutations, however mutations can be introduced efficiently at distances of at least 500 bp from the URA3 insertion. In RDL mutagenesis, to ensure that incorporation of mutations occurs at approximately equal frequencies throughout the targeted region, this region is deleted at the same time URA3 is integrated. Conclusion SSG and RDL mutagenesis allow point mutations to be easily and efficiently incorporated into the yeast genome without disrupting the native locus.

  14. Labelled agents for PET studies of the dopaminergic system -some quality assurance methods, experience and issues

    International Nuclear Information System (INIS)

    Pike, V.W.; Kensett, M.J.; Turton, D.R.; Waters, S.L.; Silvester, D.J.

    1990-01-01

    Practical methods are described for the quality assurance of three labelled agents (L-6-[ 18 F]fluoro-DOPA, S-[N-methyl- 11 C]nomifensine and [O-methyl- 11 C]raclopride) now produced regularly for PET studies of the dopaminergic system in man. These include indirect methods for the initial determination of label position (e.g. 13 C-NMR spectroscopy) and also direct methods for the assessment of chiral purity (TLC and HPLC) and the routine determination of radiochemical purity, chemical purity and specific activity (HPLC). Mass spectrometry has been used to identify some impurities. L-6-hydroxy-DOPA (a precursor in vivo of the neurotoxin, L-6-hydroxydopamine) has been detected by HPLC in some preparations of L-6-[ 18 F]fluoro-DOPA. Formulated S-[N-methyl- 11 C]nomifensine has been found to be stable. Some quality assurance issues are discussed in relation to experience in the application of the described methods and the obtained results. (author)

  15. New labeling methods via organometallic species: new synthesis of a chiral methyl group

    International Nuclear Information System (INIS)

    Faucher, Nicolas

    2000-01-01

    Chapter 1: New labeling methods via organometallic species. In the first part of this work, we have developed a new labeling strategy based on the hydrogenolysis of organolithium compounds with tritium gas or deuterium gas. This reaction is catalyzed with palladium on charcoal and leads to the labelled compounds with direct replacement of the proton by its isotopes ("2H or "3H) without further chemical modification of the target molecule. Using this strategy, tritium or deuterium atoms can be introduced in a region but also in a stereoselective fashion with more than 90% ee. The former result was obtained using (-)-sparteine during the lithiation step. Chapter II: New synthesis of a chiral methyl group. In the second part of this work, we have developed a new synthetic method to prepare chiral ditosyl-methylamine using 4,5-disubstituted oxazolidines. Dia-stereoselective substitution of the methoxy group of a 2-alkoxy-oxazolidine by a deuteride in the presence of a Lewis acid leads to the 2-deutero-oxazolidine in a highly stereoselective fashion (de = 100%). Still using a lewis acid, a tritiated hydride open the former 2-deutero-oxazolidine to afford chiral methyl group borne by the nitrogen. Further de-protection and re-protection steps lead to the ditosyl-methylamine with an ee of 65% (RIS= 83/17). Nowadays, this is the best known synthetic method, not only in terms of enantioselectivity but also in terms of chemical yield and number of radioactive steps. As NTs_2 is a fairly good leaving group, the ditosyl-methylamine offers the possibility of introducing chiral methyl group in many substrates using a S_N2 reaction with various nucleophiles. This last point leads to many potential applications in the field of biochemistry or for mechanical studies. (author) [fr

  16. Use of toxicity assessment to develop site specific remediation criteria for oil and gas facilities : guidance manual

    International Nuclear Information System (INIS)

    1996-01-01

    The results of a two year study into the evaluation of toxicity-based methods to develop site-specific, risk-based cleanup objectives for the decommissioning of oil and gas facilities were compiled into a manual of guidance. The two basic approaches used in determining remediation criteria for contaminated sites are: (1) comparison of the concentrations of chemicals found on-site with broad regional or national soil and water quality objectives developed for the chemicals involved, and (2) site-specific risk assessment. Toxicity tests are used to test organisms such as earthworms, lettuce seeds, or larval fish directly in the soil, water or sediment suspected of being contaminated. The effects of any contamination on the survival, growth, reproduction, and behaviour of the test organisms are then evaluated. The manual provides guidance in: (1) using toxicity assessments within the regulatory framework of site decommissioning, (2) performing a toxicity assessment, and (3) developing site-specific criteria for a risk assessment. 18 refs., 3 tabs., 5 figs

  17. CRISPR/Cas9-loxP-Mediated Gene Editing as a Novel Site-Specific Genetic Manipulation Tool.

    Science.gov (United States)

    Yang, Fayu; Liu, Changbao; Chen, Ding; Tu, Mengjun; Xie, Haihua; Sun, Huihui; Ge, Xianglian; Tang, Lianchao; Li, Jin; Zheng, Jiayong; Song, Zongming; Qu, Jia; Gu, Feng

    2017-06-16

    Cre-loxP, as one of the site-specific genetic manipulation tools, offers a method to study the spatial and temporal regulation of gene expression/inactivation in order to decipher gene function. CRISPR/Cas9-mediated targeted genome engineering technologies are sparking a new revolution in biological research. Whether the traditional site-specific genetic manipulation tool and CRISPR/Cas9 could be combined to create a novel genetic tool for highly specific gene editing is not clear. Here, we successfully generated a CRISPR/Cas9-loxP system to perform gene editing in human cells, providing the proof of principle that these two technologies can be used together for the first time. We also showed that distinct non-homologous end-joining (NHEJ) patterns from CRISPR/Cas9-mediated gene editing of the targeting sequence locates at the level of plasmids (episomal) and chromosomes. Specially, the CRISPR/Cas9-mediated NHEJ pattern in the nuclear genome favors deletions (64%-68% at the human AAVS1 locus versus 4%-28% plasmid DNA). CRISPR/Cas9-loxP, a novel site-specific genetic manipulation tool, offers a platform for the dissection of gene function and molecular insights into DNA-repair pathways. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

  18. Method of preparation of technetium-99m labelled radio-diagnostic agents and a stable non radio-active carrier

    International Nuclear Information System (INIS)

    1975-01-01

    A method of preparing improved technetium-99m labeled radiodiagnostic agents is described by reducing technetium-99m with stannous tartrate. Such radiodiagnostic agents are useful in scintigraphic examinations of the bone and lung

  19. Comparison of serum fractionation methods by data independent label-free proteomics

    Directory of Open Access Journals (Sweden)

    D. Baiwir

    2015-12-01

    Full Text Available Off-line sample prefractionations applied prior to biomarker discovery proteomics are options to enable more protein identifications and detect low-abundance proteins. This work compared five commercial methods efficiency to raw serum analysis using label-free proteomics. The variability of the protein quantities determined for each process was similar to the unprefractionated serum. A 49% increase in protein identifications and 12.2% of reliable quantification were obtained. A 61 times lower limit of protein quantitation was reached compared to protein concentrations observed in raw serum. The concentrations of detected proteins were confronted to estimated reference values.

  20. In silico toxicology: comprehensive benchmarking of multi-label classification methods applied to chemical toxicity data

    KAUST Repository

    Raies, Arwa B.

    2017-12-05

    One goal of toxicity testing, among others, is identifying harmful effects of chemicals. Given the high demand for toxicity tests, it is necessary to conduct these tests for multiple toxicity endpoints for the same compound. Current computational toxicology methods aim at developing models mainly to predict a single toxicity endpoint. When chemicals cause several toxicity effects, one model is generated to predict toxicity for each endpoint, which can be labor and computationally intensive when the number of toxicity endpoints is large. Additionally, this approach does not take into consideration possible correlation between the endpoints. Therefore, there has been a recent shift in computational toxicity studies toward generating predictive models able to predict several toxicity endpoints by utilizing correlations between these endpoints. Applying such correlations jointly with compounds\\' features may improve model\\'s performance and reduce the number of required models. This can be achieved through multi-label classification methods. These methods have not undergone comprehensive benchmarking in the domain of predictive toxicology. Therefore, we performed extensive benchmarking and analysis of over 19,000 multi-label classification models generated using combinations of the state-of-the-art methods. The methods have been evaluated from different perspectives using various metrics to assess their effectiveness. We were able to illustrate variability in the performance of the methods under several conditions. This review will help researchers to select the most suitable method for the problem at hand and provide a baseline for evaluating new approaches. Based on this analysis, we provided recommendations for potential future directions in this area.

  1. In silico toxicology: comprehensive benchmarking of multi-label classification methods applied to chemical toxicity data

    KAUST Repository

    Raies, Arwa B.; Bajic, Vladimir B.

    2017-01-01

    One goal of toxicity testing, among others, is identifying harmful effects of chemicals. Given the high demand for toxicity tests, it is necessary to conduct these tests for multiple toxicity endpoints for the same compound. Current computational toxicology methods aim at developing models mainly to predict a single toxicity endpoint. When chemicals cause several toxicity effects, one model is generated to predict toxicity for each endpoint, which can be labor and computationally intensive when the number of toxicity endpoints is large. Additionally, this approach does not take into consideration possible correlation between the endpoints. Therefore, there has been a recent shift in computational toxicity studies toward generating predictive models able to predict several toxicity endpoints by utilizing correlations between these endpoints. Applying such correlations jointly with compounds' features may improve model's performance and reduce the number of required models. This can be achieved through multi-label classification methods. These methods have not undergone comprehensive benchmarking in the domain of predictive toxicology. Therefore, we performed extensive benchmarking and analysis of over 19,000 multi-label classification models generated using combinations of the state-of-the-art methods. The methods have been evaluated from different perspectives using various metrics to assess their effectiveness. We were able to illustrate variability in the performance of the methods under several conditions. This review will help researchers to select the most suitable method for the problem at hand and provide a baseline for evaluating new approaches. Based on this analysis, we provided recommendations for potential future directions in this area.

  2. Occupational radiation dose assessment for a non site specific spent fuel storage facility

    International Nuclear Information System (INIS)

    Hadley, J.; Eble, R.G. Jr.

    1997-01-01

    To expedite the licensing process of the non site specific Centralized Interim Storage Facility (CISF) the Department of Energy has completed a phase I CISF Topical Safety Analysis Report (TSAR). The TSAR will be used in licensing the phase I CISF if a site is designated. An occupational radiation does assessment of the facility operations is performed as part of the phase I CISF design. The first phase of the CISF has the capability to receive, transfer, and store SNF in dual-purpose cask/canister systems (DPC's). Currently there are five vendor technologies under consideration. The preliminary dose assessment is based on estimated occupational exposures using traditional power plant ISFSI and transport cask handling processes. The second step in the process is to recommend ALARA techniques to reduce potential exposures. A final dose assessment is completed implementing the ALARA techniques and a review is performed to ensure that the design is in compliance with regulatory criteria. The dose assessment and ALARA evaluation are determined using the following input information: Dose estimates from vendor SAR's; ISFSI experience with similar systems; Traditional methods of operations; Expected CISF cask receipt rates; and feasible ALARA techniques. 5 refs., 1 tab

  3. Site Specific Effect of Tobacco Addiction in Upper Aerodigestive Tract Tumors: A Retrospective Clinicopathological Study

    Directory of Open Access Journals (Sweden)

    Ashok Kumar

    2014-01-01

    Full Text Available An institutional study was carried out in 102 patients to investigate the site specific effect of addictions, that is, tobacco smoking and tobacco chewing (smokeless, both independently and synergistically in development of malignancies in upper aerodigestive tract through retrograde questionnaire. The histopathologically proven cases were interviewed regarding different forms of addictions followed by clinical examination and investigations for grading (according to Modified Broadmann’s method and TNM staging (according to UICC according to the tumor site. Statistical analysis was done by Pearson test. Out of all proven cases of cancers, 29.4% were only tobacco chewers (smokeless, 25.5% were only smokers, 42.2% were having both types of tobacco addictions (smoke and smokeless, and only 2.9% were having no addiction. Out of only tobacco chewers (smokeless, 83.3% were of oral cavity cancers, 6.7% were of oro- and hypopharynx and the rest were of others. Among only smokers, 69.2% cases were of laryngeal and oro- and hypopharynx as compared to 11.5% of oral cavity cancers (nearly 6 times. Tobacco (smokeless chewing is associated with oral cancers whereas tobacco smoking is associated with laryngeal and hypopharyngeal carcinoma. Both smoking and smokeless tobacco act in synergy with each other.

  4. Purification and characterization of VDE, a site-specific endonuclease from the yeast Saccharomyces cerevisiae.

    Science.gov (United States)

    Gimble, F S; Thorner, J

    1993-10-15

    The 119-kDa primary translation product of the VMA1 gene of Saccharomyces cerevisiae undergoes a self-catalyzed rearrangement ("protein splicing") that excises an internal 50-kDa segment of the polypeptide and joins the amino-terminal and carboxyl-terminal segments to generate the 69-kDa subunit of the vacuolar membrane-associated H(+)-ATPase. We have shown previously that the internal segment is a site-specific endonuclease (Gimble, F. S., and Thorner, J. (1992) Nature 357, 301-306). Here we describe methods for the high level expression and purification to near homogeneity of both the authentic VMA1-derived endonuclease (or VDE) from yeast (yield 18%) and a recombinant form of VDE made in bacteria (yield 29%). Detailed characterization of these preparations demonstrated that the yeast-derived and bacterially produced enzymes were indistinguishable, as judged by: (a) behavior during purification; (b) apparent native molecular mass (50 kDa); (c) immunological reactivity; and (d) catalytic properties (specific activity; cleavage site recognition; and optima for pH, temperature, divalent cation and ionic strength). The minimal site required for VDE cleavage was delimited to a 30-base pair sequence within its specific substrate (the VMA1 delta vde allele).

  5. Sex- and Site-Specific Normative Data Curves for HR-pQCT.

    Science.gov (United States)

    Burt, Lauren A; Liang, Zhiying; Sajobi, Tolulope T; Hanley, David A; Boyd, Steven K

    2016-11-01

    The purpose of this study was to develop age-, site-, and sex-specific centile curves for common high-resolution peripheral quantitative computed tomography (HR-pQCT) and finite-element (FE) parameters for males and females older than 16 years. Participants (n = 866) from the Calgary cohort of the Canadian Multicentre Osteoporosis Study (CaMos) between the ages of 16 and 98 years were included in this study. Participants' nondominant radius and left tibia were scanned using HR-pQCT. Standard and automated segmentation methods were performed and FE analysis estimated apparent bone strength. Centile curves were generated for males and females at the tibia and radius using the generalized additive models for location, scale, and shape (GAMLSS) package in R. After GAMLSS analysis, age-, sex-, and site-specific centiles (10th, 25th, 50th, 75th, 90th) for total bone mineral density and trabecular number as well as failure load have been calculated. Clinicians and researchers can use these reference curves as a tool to assess bone health and changes in bone quality. © 2016 American Society for Bone and Mineral Research. © 2016 American Society for Bone and Mineral Research.

  6. Joint analysis of site-specific cancer risks for the atomic bomb survivors

    International Nuclear Information System (INIS)

    Pierce, D.A.; Preston, D.L.

    1993-01-01

    Statistical methods are presented for joint analysis of site-specific cancer risks for the atomic-bomb survivors. Previous analyses of these data, aside from those on leukemia, have been made either without regard to cancer type, or separately for types or classes of cancers. Clearly, analyses without regard to cancer type are less than satisfactory. The primary advantages of joint, rather than separate, analyses are that (1) models can be fitted with parameters common to cancer types, which can allow more-precise estimation of effects of interest, (2) significance tests can be used to compare type-specific risks, and (3) a clearer understanding may be obtained of risk-modification factors such as sex, age at exposure, and time since exposure. Joint analysis is straightforward, entailing primarily the incorporation of another factor for cancer type in the usual cross-tabulation of the data for analysis. The use of these methods is illustrated in an analysis of the three categories of cancer studied by the fifth Advisory Committee on the Biological Effects of Ionizing Radiation (BEIR V): digestive, respiratory, and other solid tumors. Based on this analysis, some criticism is made of the BEIR V-preferred models. Since the proposed methods are applicable to models for either relative or absolute risks, some comments on the use of explicit models for the absolute excess risk are also given. Although some of the gains from joint analysis are apparent from the results here, it will be important to use these methods with a more suitable choice of cancer classes and for cancer incidence data in which the diagnoses are more accurate. (author)

  7. Evaluation of polymorphonuclear leukocyte chemotaxis of adult and neonatal rhesus monkeys using 51-chromium labeling method

    International Nuclear Information System (INIS)

    Kinoshita, Yo; Masuda, Kiyokazu; Kobayashi, Yohnosuke

    1987-01-01

    Chemotaxis of polymorphonuclear leukocytes (PMN) from heparinized venous blood of 8 adult rhesus monkeys (Macaca Mulatta) and 13 rhesus monkey neonates within 48 hours of birth were evaluated by using 51-chromium labeling method. PMNs were prepared by Ficoll-Hypaque gradient and dextran sedimentation procedures and the final 51-chromium uptake was 3.21 ± 1.27 % to original count. PMN chemotaxis was succeeded by using two different chemotaxis filters (Nuclepore filter on top of Millipore filter) with incubation at 37 deg C for 90 min. The mean value of target: non target ratio (CPM in lower filter with chemoattractant/CPM in lower filter without chemoattractant) of 3.56 ± 2.49 from neonates showed no significant difference from that of 4.44 ± 1.24 from adults. Only about 30 % of neonates showed an impaired chemotaxis, but others showed similar chemotactic activity as adults. The results show that the 51-chromium labeling method is useful to assess neutrophil functions in rhesus monkey species and suggest that host defense mechanism of the rhesus monkey may differ from that of human in neonatal period. (author)

  8. A systematic evaluation of normalization methods in quantitative label-free proteomics.

    Science.gov (United States)

    Välikangas, Tommi; Suomi, Tomi; Elo, Laura L

    2018-01-01

    To date, mass spectrometry (MS) data remain inherently biased as a result of reasons ranging from sample handling to differences caused by the instrumentation. Normalization is the process that aims to account for the bias and make samples more comparable. The selection of a proper normalization method is a pivotal task for the reliability of the downstream analysis and results. Many normalization methods commonly used in proteomics have been adapted from the DNA microarray techniques. Previous studies comparing normalization methods in proteomics have focused mainly on intragroup variation. In this study, several popular and widely used normalization methods representing different strategies in normalization are evaluated using three spike-in and one experimental mouse label-free proteomic data sets. The normalization methods are evaluated in terms of their ability to reduce variation between technical replicates, their effect on differential expression analysis and their effect on the estimation of logarithmic fold changes. Additionally, we examined whether normalizing the whole data globally or in segments for the differential expression analysis has an effect on the performance of the normalization methods. We found that variance stabilization normalization (Vsn) reduced variation the most between technical replicates in all examined data sets. Vsn also performed consistently well in the differential expression analysis. Linear regression normalization and local regression normalization performed also systematically well. Finally, we discuss the choice of a normalization method and some qualities of a suitable normalization method in the light of the results of our evaluation. © The Author 2016. Published by Oxford University Press.

  9. Theoretical Proof and Empirical Confirmation of a Continuous Labeling Method Using Naturally 13C-Depleted Carbon Dioxide

    Institute of Scientific and Technical Information of China (English)

    Weixin Cheng; Feike A. Dijkstra

    2007-01-01

    Continuous isotope labeling and tracing is often needed to study the transformation, movement, and allocation of carbon in plant-soil systems. However, existing labeling methods have numerous limitations. The present study introduces a new continuous labeling method using naturally 13C-depleted CO2. We theoretically proved that a stable level of 13C-CO2 abundance In a labeling chamber can be maintained by controlling the rate of CO2-free air injection and the rate of ambient airflow with coupling of automatic control of CO2 concentration using a CO2 analyzer. The theoretical results were tested and confirmed in a 54 day experiment in a plant growth chamber. This new continuous labeling method avoids the use of radioactive 14C or expensive 13C-enriched CO2 required by existing methods and therefore eliminates issues of radiation safety or unaffordable isotope cost, as well as creating new opportunities for short- or long-term labeling experiments under a controlled environment.

  10. Site-Specific, Covalent Immobilization of Dehalogenase ST2570 Catalyzed by Formylglycine-Generating Enzymes and Its Application in Batch and Semi-Continuous Flow Reactors

    Directory of Open Access Journals (Sweden)

    Hui Jian

    2016-07-01

    Full Text Available Formylglycine-generating enzymes can selectively recognize and oxidize cysteine residues within the sulfatase sub motif at the terminus of proteins to form aldehyde-bearing formylglycine (FGly residues, and are normally used in protein labeling. In this study, an aldehyde tag was introduced to proteins using formylglycine-generating enzymes encoded by a reconstructed set of the pET28a plasmid system for enzyme immobilization. The haloacid dehalogenase ST2570 from Sulfolobus tokodaii was used as a model enzyme. The C-terminal aldehyde-tagged ST2570 (ST2570CQ exhibited significant enzymological properties, such as new free aldehyde groups, a high level of protein expression and improved enzyme activity. SBA-15 has widely been used as an immobilization support for its large surface and excellent thermal and chemical stability. It was functionalized with amino groups by aminopropyltriethoxysilane. The C-terminal aldehyde-tagged ST2570 was immobilized to SBA-15 by covalent binding. The site-specific immobilization of ST2570 avoided the chemical denaturation that occurs in general covalent immobilization and resulted in better fastening compared to physical adsorption. The site-specific immobilized ST2570 showed 3-fold higher thermal stability, 1.2-fold higher catalytic ability and improved operational stability than free ST2570. The site-specific immobilized ST2570 retained 60% of its original activity after seven cycles of batch operation, and it was superior to the ST2570 immobilized to SBA-15 by physical adsorption, which loses 40% of its original activity when used for the second time. It is remarkable that the site-specific immobilized ST2570 still retained 100% of its original activity after 10 cycles of reuse in the semi-continuous flow reactor. Overall, these results provide support for the industrial-scale production and application of site-specific, covalently immobilized ST2570.

  11. Comparison of a Label-Free Quantitative Proteomic Method Based on Peptide Ion Current Area to the Isotope Coded Affinity Tag Method

    Directory of Open Access Journals (Sweden)

    Young Ah Goo

    2008-01-01

    Full Text Available Recently, several research groups have published methods for the determination of proteomic expression profiling by mass spectrometry without the use of exogenously added stable isotopes or stable isotope dilution theory. These so-called label-free, methods have the advantage of allowing data on each sample to be acquired independently from all other samples to which they can later be compared in silico for the purpose of measuring changes in protein expression between various biological states. We developed label free software based on direct measurement of peptide ion current area (PICA and compared it to two other methods, a simpler label free method known as spectral counting and the isotope coded affinity tag (ICAT method. Data analysis by these methods of a standard mixture containing proteins of known, but varying, concentrations showed that they performed similarly with a mean squared error of 0.09. Additionally, complex bacterial protein mixtures spiked with known concentrations of standard proteins were analyzed using the PICA label-free method. These results indicated that the PICA method detected all levels of standard spiked proteins at the 90% confidence level in this complex biological sample. This finding confirms that label-free methods, based on direct measurement of the area under a single ion current trace, performed as well as the standard ICAT method. Given the fact that the label-free methods provide ease in experimental design well beyond pair-wise comparison, label-free methods such as our PICA method are well suited for proteomic expression profiling of large numbers of samples as is needed in clinical analysis.

  12. Doubly labeled water method: in vivo oxygen and hydrogen isotope fractionation

    International Nuclear Information System (INIS)

    Schoeller, D.A.; Leitch, C.A.; Brown, C.

    1986-01-01

    The accuracy and precision of the doubly labeled water method for measuring energy expenditure are influenced by isotope fractionation during evaporative water loss and CO 2 excretion. To characterize in vivo isotope fractionation, we collected and isotopically analyzed physiological fluids and gases. Breath and transcutaneous water vapor were isotopically fractionated. The degree of fractionation indicated that the former was fractionated under equilibrium control at 37 0 C, and the latter was kinetically fractionated. Sweat and urine were unfractionated. By use of isotopic balance models, the fraction of water lost via fractionating routes was estimated from the isotopic abundances of body water, local drinking water, and dietary solids. Fractionated water loss averaged 23% (SD = 10%) of water turnover, which agreed with our previous estimates based on metabolic rate, but there was a systematic difference between the results based on O 2 and hydrogen. Corrections for isotopic fractionation of water lost in breath and (nonsweat) transcutaneous loss should be made when using labeled water to measure water turnover or CO 2 production

  13. Study on labelling conditions of 125I-synkavit by the iodogen method

    International Nuclear Information System (INIS)

    Oezdemir, D.; Uenak, P.

    1994-01-01

    Labeling conditions of synkavit (2-methyl-1,4-naphtoquinol disodium phosphate) with iodine-125 have been studied. Labeling temperature, reaction time, successive using of iodogen coated tubes, iodogen amount and synkavit concentrations have been determined to get optimum conditions for maximum labeling. Final results showed that when the labeling temperature, reaction time, synkavit concentration and iodogen amount were, at room temperature, 15 min (in the case of successive using of three iodogen coated tubes), 2 mg ml -1 and 5 mg, respectively; labeling yield was 90% and specific activities of the order of 555 GBq mmol -1 (15 Ci mmol -1 have been obtained. (author) 14 refs.; 4 figs

  14. Site-Specific Atmospheric Dispersion Characteristics of Korean Nuclear Power Plant Sites

    International Nuclear Information System (INIS)

    Han, M. H.; Kim, E. H.; Suh, K. S.; Hwang, W. T.; Choi, Y. G.

    2001-01-01

    Site-specific atmospheric dispersion characteristics have been analyzed. The northwest and the southwest wind prevail on nuclear sites of Korea. The annual isobaric surface averaged for twenty years around Korean peninsula shows that west wind prevails. The prevailing west wind is profitable in the viewpoint of radiation protection because three of four nuclear sites are located in the east side. Large scale field tracer experiments over nuclear sites have been conducted for the purpose of analyzing the atmospheric dispersion characteristics and validating a real-time atmospheric dispersion and dose assessment system FADAS. To analyze the site-specific atmospheric dispersion characteristics is essential for making effective countermeasures against a nuclear emergency

  15. Drainage filter technologies to mitigate site-specific phosphorus losses in agricultural drainage discharge

    DEFF Research Database (Denmark)

    Kjærgaard, Charlotte; Heckrath, Goswin Johann; Canga, Eriona

    in drainage. The Danish “SUPREME-TECH” project (2010-2016) (www.supreme-tech.dk) aims at providing the scientific basis for developing cost-effective filter technologies for P in agricultural drainage waters. The project studies different approaches of implementing filter technologies including drainage well....... Targeting high risk areas of P loss and applying site-specific measures promises to be a cost-efficient approach. The Danish Commission for Nature and Agriculture has, therefore, now called for a paradigm shift towards targeted, cost-efficient technologies to mitigate site-specific nutrient losses...... environmental threshold values (

  16. Accounting for both local aquatic community composition and bioavailability in setting site-specific quality standards for zinc.

    Science.gov (United States)

    Peters, Adam; Simpson, Peter; Moccia, Alessandra

    2014-01-01

    Recent years have seen considerable improvement in water quality standards (QS) for metals by taking account of the effect of local water chemistry conditions on their bioavailability. We describe preliminary efforts to further refine water quality standards, by taking account of the composition of the local ecological community (the ultimate protection objective) in addition to bioavailability. Relevance of QS to the local ecological community is critical as it is important to minimise instances where quality classification using QS does not reconcile with a quality classification based on an assessment of the composition of the local ecology (e.g. using benthic macroinvertebrate quality assessment metrics such as River InVertebrate Prediction and Classification System (RIVPACS)), particularly where ecology is assessed to be at good or better status, whilst chemical quality is determined to be failing relevant standards. The alternative approach outlined here describes a method to derive a site-specific species sensitivity distribution (SSD) based on the ecological community which is expected to be present at the site in the absence of anthropogenic pressures (reference conditions). The method combines a conventional laboratory ecotoxicity dataset normalised for bioavailability with field measurements of the response of benthic macroinvertebrate abundance to chemical exposure. Site-specific QSref are then derived from the 5%ile of this SSD. Using this method, site QSref have been derived for zinc in an area impacted by historic mining activities. Application of QSref can result in greater agreement between chemical and ecological metrics of environmental quality compared with the use of either conventional (QScon) or bioavailability-based QS (QSbio). In addition to zinc, the approach is likely to be applicable to other metals and possibly other types of chemical stressors (e.g. pesticides). However, the methodology for deriving site-specific targets requires

  17. SU-F-BRD-07: Empirical Derivation of Site-Specific Margin Formulas

    International Nuclear Information System (INIS)

    Conroy, L; Smith, W; Quirk, S

    2014-01-01

    systematic error. Site-specific margin formulas may be better suited to patient populations than theoretically derived, ideal geometry general margin formulas

  18. Global site-specific analysis of glycoprotein N-glycan processing.

    Science.gov (United States)

    Cao, Liwei; Diedrich, Jolene K; Ma, Yuanhui; Wang, Nianshuang; Pauthner, Matthias; Park, Sung-Kyu Robin; Delahunty, Claire M; McLellan, Jason S; Burton, Dennis R; Yates, John R; Paulson, James C

    2018-06-01

    N-glycans contribute to the folding, stability and functions of the proteins they decorate. They are produced by transfer of the glycan precursor to the sequon Asn-X-Thr/Ser, followed by enzymatic trimming to a high-mannose-type core and sequential addition of monosaccharides to generate complex-type and hybrid glycans. This process, mediated by the concerted action of multiple enzymes, produces a mixture of related glycoforms at each glycosite, making analysis of glycosylation difficult. To address this analytical challenge, we developed a robust semiquantitative mass spectrometry (MS)-based method that determines the degree of glycan occupancy at each glycosite and the proportion of N-glycans processed from high-mannose type to complex type. It is applicable to virtually any glycoprotein, and a complete analysis can be conducted with 30 μg of protein. Here, we provide a detailed description of the method that includes procedures for (i) proteolytic digestion of glycoprotein(s) with specific and nonspecific proteases; (ii) denaturation of proteases by heating; (iii) sequential treatment of the glycopeptide mixture with two endoglycosidases, Endo H and PNGase F, to create unique mass signatures for the three glycosylation states; (iv) LC-MS/MS analysis; and (v) data analysis for identification and quantitation of peptides for the three glycosylation states. Full coverage of site-specific glycosylation of glycoproteins is achieved, with up to thousands of high-confidence spectra hits for each glycosite. The protocol can be performed by an experienced technician or student/postdoc with basic skills for proteomics experiments and takes ∼7 d to complete.

  19. Application of site-specific natural isotope fractionation (SNIF-NMR) of hydrogen to the characterization of European beers

    International Nuclear Information System (INIS)

    Martin, G.J.; Benbernou, M.; Lantier, F.

    1985-01-01

    More than one hundred samples of European beers have been investigated by the new SNIF-NMR method which is based on 2H NMR at the natural abundance level and enables site-specific natural isotope fractionation factors in ethanol to be determined. The relative (R) and absolute (D/H)sub(i) parameters are shown to be characteristic of the country where the beers are brewed and the observed variations are explained in terms of cereal composition, water resource, and manufacturing processing (fermentation, yeast, temperature cycle). These new parameters find analytical and mechanistic applications in the identification of a beer and in the investigation of a fermentation process. (author)

  20. Development of methods for the purification of 67Ga and 68Ga for biomolecules labeling

    International Nuclear Information System (INIS)

    Costa, Renata Ferreira

    2012-01-01

    For more than fifty years, the long-lived 68 Ge/ 68 Ga generators have been in development, obtaining 68 Ga without the need of having in house cyclotron, which is a considerable convenience for PET centers that have no nearby cyclotrons. 68 Ga decays 89% by positron emission and low photon emission (1077 keV) and the physical half life of 67.7 minutes is compatible with the pharmacokinetics of low biomolecular weight substances like peptides and antibody fragments. Moreover, its established metallic chemistry allows it to be stably bound to the carrier peptide sequence via a suitable bifunctional chelator, such as DOTA. All these reasons together with the technology of PET/CT allowed advances in molecular imaging, in particular in the diagnosis of neuroendocrine diseases. However, the eluate from the commercial 68 Ge/ 68 Ga generators still contains high levels of long lived 68 Ge, besides other metallic impurities, which competes with 68 Ga with a consequent reduction of the labeling yield of biomolecules, such as Fe 3+ and Zn 2+ . Thus, the lower the amount of impurities in the eluate, the competition between the radiolabeled and unlabeled peptide by the receptor will be smaller and the quality of imaging will be better, a subsequent purification step is needed after the generator elution. The aim of this work is to evaluate different purifications methods of 68 Ga to label biomolecules, with emphasis on the study of the chemical impurities contained in the eluate and to develop a new purification method. Several purification methods were studied. Many cationic resin were tested simulating the commercial process. 68 Ga is adsorbed in cationic resin, which is not commercial available and eluted in acid/acetone solution. The use of minor particles of cationic resin AG50W-X4 (200-400 mesh) showed the best results. An innovate method was the extraction chromatography, which is based on the absorption of diisopropyl ether in XAD 16 and 68 Ga recovery in deionized

  1. Overexpression of a homogeneous oligosaccharide with {sup 13}C labeling by genetically engineered yeast strain

    Energy Technology Data Exchange (ETDEWEB)

    Kamiya, Yukiko; Yamamoto, Sayoko [National Institutes of Natural Sciences, Okazaki Institute for Integrative Bioscience and Institute for Molecular Science (Japan); Chiba, Yasunori; Jigami, Yoshifumi [National Institute of Advanced Industrial Science and Technology, Research Center for Medical Glycoscience (Japan); Kato, Koichi, E-mail: kkatonmr@ims.ac.jp [National Institutes of Natural Sciences, Okazaki Institute for Integrative Bioscience and Institute for Molecular Science (Japan)

    2011-08-15

    This report describes a novel method for overexpression of {sup 13}C-labeled oligosaccharides using genetically engineered Saccharomyces cerevisiae cells, in which a homogeneous high-mannose-type oligosaccharide accumulates because of deletions of genes encoding three enzymes involved in the processing pathway of asparagine-linked oligosaccharides in the Golgi complex. Using uniformly {sup 13}C-labeled glucose as the sole carbon source in the culture medium of these engineered yeast cells, high yields of the isotopically labeled Man{sub 8}GlcNAc{sub 2} oligosaccharide could be successfully harvested from glycoprotein extracts of the cells. Furthermore, {sup 13}C labeling at selected positions of the sugar residues in the oligosaccharide could be achieved using a site-specific {sup 13}C-enriched glucose as the metabolic precursor, facilitating NMR spectral assignments. The {sup 13}C-labeling method presented provides the technical basis for NMR analyses of structures, dynamics, and interactions of larger, branched oligosaccharides.

  2. Determination of blood circulation in oral formations using Rb86 distribution method and labelled micropearl method

    International Nuclear Information System (INIS)

    Fazekas, A.; Posch, E.; Harsing, L.

    1979-01-01

    The blood circulation of incisors, dental pulp and tongue was detemined using the measurement of 86 Rb distribution in rats. The results were compared with those obtained by a simultaneous micropearl method. It was found that 37 per cent of 86 Rb in dental tissues is localized in the hard propiodentium, with a high proportion diffusing from the periodontium. The 86 Rb fraction localized in the tongue represents its blood circulation. (author)

  3. The method for production of high purity carrier free ortophosphoric acid labeled with isotopes Phosphorus-32 and Phosphorus-33

    International Nuclear Information System (INIS)

    Abdukayumov, M.N.; Abdusalyamov, A.N.; Chistyakov, P.G.; Yuldashev, B.S.

    2001-01-01

    Extensive application for various radioactive isotopes was found in an extremity of the 20-Th century in a science and production. Labeled compounds are used with growing effectiveness in a molecular biology, gene engineering, medicine and other areas. Phosphorus-32 and Phosphorus-33 isotopes as a different labeled compounds that are used mainly in molecular biology are produced at the Radiopreparat enterprise of the Institute of Nuclear Physics of Academy of Sciences of Uzbekistan Republic. The quality of labeled preparations is very high. The specifications for above mentioned preparations corresponds to demands most of customers in different countries. P-32 or P-33 labeled orthophosphoric acid has high radiochemical purity (more than 99 %) and specific radioactivity close to theoretical. Orthophosphoric acid prepared by the described above method has radiochemical purity about 95 % and output of the target product 99%

  4. Method of preparing thymidine-5'-monophosphate specifically or nonspecifically labelled with 14C or with 3H

    International Nuclear Information System (INIS)

    Nejedly, Z.; Filip, J.; Ekl, J.; Kolina, J.; Votruba, I.; Skoda, J.

    1977-01-01

    The invention claims a method for labelled thymidine-5'-monophosphate preparation by cultivating a special thymine-dependent Escherichia coli SPT - strain in the optimum synthetic culture medium containing 0.8 to 1.2 g/ml of labelled thymine. Practically the whole amount of labelled thymine is utilized for cellular deoxyribonucleic acid synthesis. The radioactive biomass obtained is processed using such chemical and enzymatic decomposition procedures as to allow separating the labelled thymidine-5'-monophosphate as the only thymine reaction product. Experiments conducted showed that the radiochemical purity of the thymidine-5'-monophosphate obtained was better than 98%. The absence of other nonactive substances was confirmed by spectrophotometric analysis. The overall product activity was 92.3% of the activity of thymine-2- 14 C introduced in the reaction. (Ha)

  5. A new biological method for preparing certain sulphurated substances labelled with S35

    International Nuclear Information System (INIS)

    Chapeville, F.; Maier-Huser, H.; Fromageot, P.

    1962-01-01

    Previous investigations have shown that the yolk-sac of embryonic bird's eggs can be used to produce the following reactions: (a) reduction of sulphate to sulphite; (b) fixation of the sulphite on the carbon chain produced by the desulf-hydration of l-cysteine, with formation of l-cysteic acid; (c) decarboxylation of the l-cysteine acid into taurine. The enzymatic system which causes reaction (b) has been purified. It also acts as a catalyst in the sulphur-exchange between the cysteine and the mineral sulphide. The authors have utilized these data in preparing sulphurated substances labelled with S 35 : taurine S 35 , l-cysteine S 35 and l-cysteic acid S 35 . For each of the three, they discuss the chemical reactions involved, the methods of preparation, the experimental conditions of extraction and purity-control, together with the yields and specific activities obtained. (authors) [fr

  6. A modified method for the in vivo labeling of red blood cells with /sup 99m/Tc: concise communication

    International Nuclear Information System (INIS)

    Callahan, R.J.; Froelich, J.W.; McKusick, K.A.; Leppo, J.; Strauss, H.W.

    1982-01-01

    The rate of incorporation of /sup 99m/Tc into red blood cells pretinned in vivo was measured by collecting blood samples in stannous DTPA solution, which served as a competing ligand for /sup 99m/Tc. This collection technique permitted a measurement of high-affinity red-cell labeling efficiency at the instant of sampling. At 0.5 min after injection only 62% of technetium is tightly bound to the red cell; this rises to 94.5% at 10 min. Based on the graded labeling of the red cells, the in vivo labeling procedure was modified by isolating pertechnetate and red blood cells tinned in vivo in a syringe during the first 10 min of labeling. The pertechnetate is thus prevented from distributing to extravascular compartments, and 90% of the injected /sup 99m/Tc is firmly bound to red blood cells at the time of injection. In a series of 23 patients, seven were tested with the in vivo method and seven with the modified in vivo method, and nine patients were tested with each method on separate occasions. A decrease in gastric activity and improved image quality were found with the modified method compared with the standard method of in vivo red-cell labeling

  7. Evaluating the results of a site-specific PSHA from the perspective of a risk analyst

    Science.gov (United States)

    Klügel, Jens-Uwe

    2016-04-01

    From 1998 till 2015 Swiss Nuclear Power Plants sponsored a set of comprehensive site-specific PSHA-studies (PEGASOS, PEGASOS Refinement Project) to obtain the requested input for their plant specific probabilistic risk assessments following the US SSHAC procedures at their most elaborated level 4. The studies were performed by well-known earth scientists working completely independent from sponsors under participatory review of the Swiss Nuclear Safety Inspectorate. Risk analysts of Swiss Nuclear Power Plants recently have been mandated to implement the final results of the studies in their risk assessment studies. This triggered an in depth assessment of the results focussed on their practical applicability for risk studies. This assessment resulted in some important insights that are of interest for future PSHA studies performed for new nuclear power plants. The assessment included a review of the completeness of results with respect to risk applications as well as plausibility checks of hazard results based on Black Swan Theory and known historical events. The key lessons and recommendations for more detailed project output specifications for future projects are presented in the paper. It was established that future PSHA projects shall provide the joint probability distribution of ground motion hazard and the associated strong motion duration as the output to allow for a technically meaningful risk assessment. The recommendation of WENRA (West European Nuclear Regulators) published in their reference levels to perform natural hazard assessment preferably based on physical grounds (deterministic method) is also rationalized by recommending an holistic approach to hazard analysis comparing PSHA insights with the results of modelling deterministic Seismic Hazard Analysis.

  8. Structural basis for the site-specific incorporation of lysine derivatives into proteins.

    Directory of Open Access Journals (Sweden)

    Veronika Flügel

    Full Text Available Posttranslational modifications (PTMs of proteins determine their structure-function relationships, interaction partners, as well as their fate in the cell and are crucial for many cellular key processes. For instance chromatin structure and hence gene expression is epigenetically regulated by acetylation or methylation of lysine residues in histones, a phenomenon known as the 'histone code'. Recently it was shown that these lysine residues can furthermore be malonylated, succinylated, butyrylated, propionylated and crotonylated, resulting in significant alteration of gene expression patterns. However the functional implications of these PTMs, which only differ marginally in their chemical structure, is not yet understood. Therefore generation of proteins containing these modified amino acids site specifically is an important tool. In the last decade methods for the translational incorporation of non-natural amino acids using orthogonal aminoacyl-tRNA synthetase (aaRS:tRNAaaCUA pairs were developed. A number of studies show that aaRS can be evolved to use non-natural amino acids and expand the genetic code. Nevertheless the wild type pyrrolysyl-tRNA synthetase (PylRS from Methanosarcina mazei readily accepts a number of lysine derivatives as substrates. This enzyme can further be engineered by mutagenesis to utilize a range of non-natural amino acids. Here we present structural data on the wild type enzyme in complex with adenylated ε-N-alkynyl-, ε-N-butyryl-, ε-N-crotonyl- and ε-N-propionyl-lysine providing insights into the plasticity of the PylRS active site. This shows that given certain key features in the non-natural amino acid to be incorporated, directed evolution of this enzyme is not necessary for substrate tolerance.

  9. 76 FR 24831 - Site-Specific Analyses for Demonstrating Compliance With Subpart C Performance Objectives

    Science.gov (United States)

    2011-05-03

    ...-level radioactive waste disposal facilities to conduct site-specific analyses to demonstrate compliance... public health and safety, these amendments would enhance the safe disposal of low-level radioactive waste... would be to enhance the safe disposal of low-level radioactive waste. The NRC is also proposing...

  10. Hellsgate Big Game Winter Range Wildlife Mitigation Site Specific Management Plan for the Hellsgate Project.

    Energy Technology Data Exchange (ETDEWEB)

    Berger, Matthew T.; Judd, Steven L.

    1999-01-01

    This report contains a detailed site-specific management plan for the Hellsgate Winter Range Wildlife Mitigation Project. The report provides background information about the mitigation process, the review process, mitigation acquisitions, Habitat Evaluation Procedures (HEP) and mitigation crediting, current habitat conditions, desired future habitat conditions, restoration/enhancements efforts and maps.

  11. 30 CFR 46.11 - Site-specific hazard awareness training.

    Science.gov (United States)

    2010-07-01

    ... Section 46.11 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR EDUCATION AND... workers; (4) Customers, including commercial over-the-road truck drivers; (5) Construction workers or... procedures. The training must address site-specific health and safety risks, such as unique geologic or...

  12. Environmental Restoration Site-Specific Plan for the Portsmouth Gaseous Diffusion Plant, FY 93

    International Nuclear Information System (INIS)

    1993-01-01

    The purpose of this Site-Specific Plan (SSP) is to describe past, present, and future activities undertaken to implement Environmental Restoration and Waste Management goals at the Portsmouth Gaseous Diffusion Plant (PORTS). The SSP is presented in sections emphasizing Environmental Restoration description of activities, resources, and milestones

  13. 75 FR 64718 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2010-10-20

    ... Test Site including decontamination, closure, re-use and/or demolition. Purpose of the Soils Committee: The purpose of the Committee is to focus on issues related to soil contamination at the Nevada Test... Industrial Sites and Soils Committees of the Environmental Management Site-Specific Advisory Board (EM SSAB...

  14. 76 FR 5365 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2011-01-31

    ... Industrial Sites and Soils Committees of the Environmental Management Site-Specific Advisory Board (EM SSAB... sites at the Nevada National Security Site including decontamination, closure, re-use and/or demolition. Purpose of the Soils Committee: The purpose of the Committee is to focus on issues related to soil...

  15. 75 FR 71677 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2010-11-24

    ... Industrial Sites and Soils Committees of the Environmental Management Site-Specific Advisory Board (EM SSAB... sites at the Nevada Test Site including decontamination, closure, re-use and/or demolition. Purpose of the Soils Committee: The purpose of the Committee is to focus on issues related to soil contamination...

  16. Deconstructing thermodynamic parameters of a coupled system from site-specific observables.

    Science.gov (United States)

    Chowdhury, Sandipan; Chanda, Baron

    2010-11-02

    Cooperative interactions mediate information transfer between structural domains of a protein molecule and are major determinants of protein function and modulation. The prevalent theories to understand the thermodynamic origins of cooperativity have been developed to reproduce the complex behavior of a global thermodynamic observable such as ligand binding or enzyme activity. However, in most cases the measurement of a single global observable cannot uniquely define all the terms that fully describe the energetics of the system. Here we establish a theoretical groundwork for analyzing protein thermodynamics using site-specific information. Our treatment involves extracting a site-specific parameter (defined as χ value) associated with a structural unit. We demonstrate that, under limiting conditions, the χ value is related to the direct interaction terms associated with the structural unit under observation and its intrinsic activation energy. We also introduce a site-specific interaction energy term (χ(diff)) that is a function of the direct interaction energy of that site with every other site in the system. When combined with site-directed mutagenesis and other molecular level perturbations, analyses of χ values of site-specific observables may provide valuable insights into protein thermodynamics and structure.

  17. Integration of aerial imaging and variable-rate technology for site-specific aerial herbicide application

    Science.gov (United States)

    As remote sensing and variable rate technology are becoming more available for aerial applicators, practical methodologies on effective integration of these technologies are needed for site-specific aerial applications of crop production and protection materials. The objectives of this study were to...

  18. Microbial profile comparisons of saliva, pooled and site-specific subgingival samples in periodontitis patients

    DEFF Research Database (Denmark)

    Belstrøm, Daniel; Sembler-Møller, Maria Lynn; Grande, Maria Anastasia

    2017-01-01

    by pooled subgingival samples. Presence of Porphyromonas gingivalis, Treponema denticola, Prevotella intermedia, Filifactor alocis, Tannerella forsythia and Parvimona micra in site-specific subgingival samples were detected in saliva with an AUC of 0.79 (sensitivity: 0.61, specificity: 0.94), compared...

  19. Potentials for site-specific design of MW sized wind turbines

    DEFF Research Database (Denmark)

    Thomsen, K.; Fuglsang, P.; Schepers, G.

    2001-01-01

    The potential for site specific design of MW sized wind turbines is quantified by comparing design loads for wind turbines installed at a range of different sites. The sites comprise on-shore normal flat terrain stand-alone conditions and wind farm conditions together with offshore and mountainous...

  20. 78 FR 26005 - Environmental Management Site-Specific Advisory Board, Savannah River Site

    Science.gov (United States)

    2013-05-03

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Savannah River Site. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  1. 78 FR 73519 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2013-12-06

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  2. 78 FR 45518 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2013-07-29

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  3. 78 FR 58294 - Environmental Management Site-Specific Advisory Board, Idaho National Laboratory

    Science.gov (United States)

    2013-09-23

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Idaho National Laboratory. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  4. 78 FR 38969 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2013-06-28

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  5. 78 FR 23760 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2013-04-22

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Nevada. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  6. 78 FR 61348 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2013-10-03

    ...On September 16, 2013, in FR Doc. 2013-22453, on page 56871, the Department of Energy (DOE) published a notice of open meeting announcing a meeting on October 2, 2013 of the Environmental Management Site-Specific Advisory Board, Portsmouth (78 FR 56871). This notice announces the cancellation of this meeting.

  7. 78 FR 65979 - Environmental Management Site-Specific Advisory Board, Savannah River Site

    Science.gov (United States)

    2013-11-04

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Savannah River Site. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  8. 77 FR 50488 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2012-08-21

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  9. 78 FR 12746 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Science.gov (United States)

    2013-02-25

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  10. 78 FR 40130 - Environmental Management Site-Specific Advisory Board, Savannah River Site

    Science.gov (United States)

    2013-07-03

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Savannah River Site. The Federal Advisory Committee Act (Pub. L. No. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  11. 78 FR 75552 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Science.gov (United States)

    2013-12-12

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  12. 77 FR 65374 - Environmental Management Site-Specific Advisory Board, Idaho National Laboratory

    Science.gov (United States)

    2012-10-26

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Idaho National Laboratory. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  13. 78 FR 17192 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2013-03-20

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Nevada. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  14. 78 FR 44942 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Science.gov (United States)

    2013-07-25

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  15. 77 FR 31837 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2012-05-30

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act (Pub. L. No. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  16. 78 FR 63172 - Environmental Management Site-Specific Advisory Board, Paducah; Meeting

    Science.gov (United States)

    2013-10-23

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  17. 77 FR 2713 - Environmental Management Site-Specific Advisory Board, Hanford

    Science.gov (United States)

    2012-01-19

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Hanford. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  18. 77 FR 26273 - Environmental Management Site-Specific Advisory Board, Northern New Mexico

    Science.gov (United States)

    2012-05-03

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Northern New Mexico. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  19. 77 FR 24695 - Environmental Management Site-Specific Advisory Board, Savannah River Site

    Science.gov (United States)

    2012-04-25

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Savannah River Site. The Federal Advisory Committee Act (Pub. L. . 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  20. 77 FR 55813 - Environmental Management Site-Specific Advisory Board Chairs

    Science.gov (United States)

    2012-09-11

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB) Chairs. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  1. 77 FR 47047 - Environmental Management Site-Specific Advisory Board, Northern New Mexico

    Science.gov (United States)

    2012-08-07

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Northern New Mexico. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  2. 78 FR 49738 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Science.gov (United States)

    2013-08-15

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  3. 78 FR 64208 - Environmental Management Site-Specific Advisory Board Chairs

    Science.gov (United States)

    2013-10-28

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB) Chairs. The Federal Advisory Committee Act (Pub. L. No. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  4. 77 FR 28368 - Environmental Management Site-Specific Advisory Board, Hanford

    Science.gov (United States)

    2012-05-14

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Hanford. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  5. 77 FR 18243 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Science.gov (United States)

    2012-03-27

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  6. 78 FR 54460 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2013-09-04

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  7. 77 FR 60688 - Environmental Management Site-Specific Advisory Board, Savannah River Site

    Science.gov (United States)

    2012-10-04

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Savannah River Site. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  8. 78 FR 17648 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Science.gov (United States)

    2013-03-22

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  9. 77 FR 18242 - Environmental Management Site-Specific Advisory Board Chairs

    Science.gov (United States)

    2012-03-27

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB) Chairs. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  10. 78 FR 23241 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Science.gov (United States)

    2013-04-18

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  11. 78 FR 46330 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2013-07-31

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Nevada. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  12. 78 FR 25064 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2013-04-29

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  13. 78 FR 38305 - Environmental Management Site-Specific Advisory Board, Northern New Mexico

    Science.gov (United States)

    2013-06-26

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Northern New Mexico. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  14. 78 FR 69657 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2013-11-20

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  15. 77 FR 49442 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Science.gov (United States)

    2012-08-16

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  16. 77 FR 13104 - Environmental Management Site-Specific Advisory Board, Savannah River Site

    Science.gov (United States)

    2012-03-05

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Savannah River Site. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  17. 77 FR 53192 - Environmental Management Site-Specific Advisory Board, Idaho National Laboratory

    Science.gov (United States)

    2012-08-31

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Idaho National Laboratory. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  18. 77 FR 39235 - Environmental Management Site-Specific Advisory Board, Savannah River Site

    Science.gov (United States)

    2012-07-02

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Savannah River Site. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  19. 77 FR 58364 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Science.gov (United States)

    2012-09-20

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  20. 78 FR 78952 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2013-12-27

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  1. 77 FR 16021 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2012-03-19

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  2. 78 FR 16260 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2013-03-14

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act (Pub. L. No. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  3. 77 FR 76475 - Environmental Management Site-Specific Advisory Board, Northern New Mexico

    Science.gov (United States)

    2012-12-28

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Northern New Mexico. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  4. 78 FR 30911 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Science.gov (United States)

    2013-05-23

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub. L. No. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  5. 78 FR 22255 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2013-04-15

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  6. 78 FR 716 - Environmental Management Site-Specific Advisory Board, Savannah River Site

    Science.gov (United States)

    2013-01-04

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Savannah River Site. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  7. 78 FR 32640 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2013-05-31

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  8. 77 FR 45345 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Science.gov (United States)

    2012-07-31

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  9. 77 FR 24694 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2012-04-25

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Nevada. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  10. 78 FR 49739 - Environmental Management Site-Specific Advisory Board, Northern New Mexico

    Science.gov (United States)

    2013-08-15

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Northern New Mexico. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  11. 78 FR 16260 - Environmental Management Site-Specific Advisory Board, Savannah River Site

    Science.gov (United States)

    2013-03-14

    ...On March 4, 2013, the Department of Energy (DOE) published a notice of open meeting announcing a meeting on March 25-26, 2013 of the Environmental Management Site-Specific Advisory Board, Savannah River Site (78 FR 14088). This document makes a correction to that notice.

  12. 78 FR 10611 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2013-02-14

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  13. 78 FR 68431 - Environmental Management Site-Specific Advisory Board, Hanford

    Science.gov (United States)

    2013-11-14

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Hanford. The Federal Advisory Committee Act (Pub. L. No. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  14. 78 FR 54461 - Environmental Management Site-Specific Advisory Board, Savannah River Site

    Science.gov (United States)

    2013-09-04

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Savannah River Site. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  15. 77 FR 2714 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Science.gov (United States)

    2012-01-19

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  16. 77 FR 4799 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2012-01-31

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  17. 77 FR 53193 - Environmental Management Site-Specific Advisory Board, Savannah River Site

    Science.gov (United States)

    2012-08-31

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Savannah River Site. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  18. 78 FR 26635 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2013-05-07

    ...On April 29, 2013, the Department of Energy (DOE) published a notice of open meeting announcing a meeting on May 16, 2013 of the Environmental Management Site-Specific Advisory Board, Paducah (78 FR 25064). This document makes a correction to that notice.

  19. 78 FR 7767 - Environmental Management Site-Specific Advisory Board, Paducah

    Science.gov (United States)

    2013-02-04

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Paducah. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  20. 78 FR 53135 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2013-08-28

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Nevada. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  1. 77 FR 49442 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2012-08-16

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Nevada. The Federal Advisory Committee Act (Pub. L. No. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  2. 77 FR 63300 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2012-10-16

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  3. 78 FR 3890 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Science.gov (United States)

    2013-01-17

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  4. 78 FR 4139 - Environmental Management Site-Specific Advisory Board, Hanford

    Science.gov (United States)

    2013-01-18

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Hanford. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  5. 78 FR 20311 - Environmental Management Site-Specific Advisory Board Chairs

    Science.gov (United States)

    2013-04-04

    ...This notice announces a webinar of the Environmental Management Site-Specific Advisory Board (EM SSAB) Chairs. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this webinar be announced in the Federal Register.

  6. 77 FR 64112 - Environmental Management Site-Specific Advisory Board, Hanford

    Science.gov (United States)

    2012-10-18

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Hanford. The Federal Advisory Committee Act (Pub. L. No. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  7. 78 FR 63171 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation; Meeting

    Science.gov (United States)

    2013-10-23

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  8. 77 FR 39234 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2012-07-02

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Nevada. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  9. 78 FR 58292 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Science.gov (United States)

    2013-09-23

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  10. 77 FR 74836 - Environmental Management Site-Specific Advisory Board, Oak Ridge Reservation

    Science.gov (United States)

    2012-12-18

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Oak Ridge Reservation. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  11. 77 FR 22566 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2012-04-16

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  12. 78 FR 64932 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2013-10-30

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Nevada. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  13. 78 FR 63171 - Environmental Management Site-Specific Advisory Board, Northern New Mexico; Meeting

    Science.gov (United States)

    2013-10-23

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Northern New Mexico. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  14. 78 FR 28207 - Environmental Management Site-Specific Advisory Board, Hanford

    Science.gov (United States)

    2013-05-14

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Hanford. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  15. 78 FR 49738 - Environmental Management Site-Specific Advisory Board, Hanford

    Science.gov (United States)

    2013-08-15

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Hanford. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  16. 77 FR 39234 - Environmental Management Site-Specific Advisory Board, Northern New Mexico

    Science.gov (United States)

    2012-07-02

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Northern New Mexico. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  17. 78 FR 30910 - Environmental Management Site-Specific Advisory Board, Idaho National Laboratory

    Science.gov (United States)

    2013-05-23

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Idaho National Laboratory. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  18. 78 FR 36543 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2013-06-18

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  19. 77 FR 16021 - Environmental Management Site-Specific Advisory Board, Nevada

    Science.gov (United States)

    2012-03-19

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Nevada. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  20. 78 FR 23759 - Environmental Management Site-Specific Advisory Board, Northern New Mexico

    Science.gov (United States)

    2013-04-22

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Northern New Mexico. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  1. 78 FR 59012 - Environmental Management Site-Specific Advisory Board Chairs

    Science.gov (United States)

    2013-09-25

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB) Chairs. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  2. 78 FR 56871 - Environmental Management Site-Specific Advisory Board, Portsmouth

    Science.gov (United States)

    2013-09-16

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Portsmouth. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  3. 77 FR 76475 - Environmental Management Site-Specific Advisory Board, Idaho National Laboratory

    Science.gov (United States)

    2012-12-28

    ...This notice announces a meeting of the Environmental Management Site-Specific Advisory Board (EM SSAB), Idaho National Laboratory. The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  4. 76 FR 18921 - Land Disposal Restrictions: Nevada and California; Site Specific Treatment Variances for...

    Science.gov (United States)

    2011-04-06

    ... final actions to both issue a site- specific treatment variance to U.S. Ecology Nevada (USEN) in Beatty... Facility is open from 8:30 a.m. to 4:30 p.m., Monday through Friday, excluding legal holidays. The... action and anticipate no adverse comment. Based on the information and data submitted by the petitioner...

  5. Creating prescription maps from satellite imagery for site-specific management of cotton root rot

    Science.gov (United States)

    Cotton root rot is a century-old cotton disease that can now be controlled with Topguard Terra Fungicide. However, as this disease tends to occur in the same general areas within fields year after year, site-specific treatment can be more effective and economical. The objective of this study was to ...

  6. Site-specific management of cotton root rot using historical remote sensing imagery

    Science.gov (United States)

    Cotton root rot can now be effectively controlled with Topguard Terra Fungicide, but site-specific application of the fungicide can greatly reduce treatment cost as only portions of the field are infested with the disease. The overall goal of this three-year project was to demonstrate how to use his...

  7. Summary of some feasibility studies for site-specific solar industrial process heat

    Energy Technology Data Exchange (ETDEWEB)

    None

    1982-01-01

    Some feasibility studies for several different site specific solar industrial process heat applications are summarized. The followng applications are examined. Leather Tanning; Concrete Production: Lumber and Paper Processing; Milk Processing; Molding, Curing or Drying; Automobile Manufacture; and Food Processing and Preparation. For each application, site and process data, system design, and performance and cost estimates are summarized.

  8. Site Specific Waste Management Instruction for the 116-F-4 soil storage unit

    International Nuclear Information System (INIS)

    Hopkins, G.G.

    1996-08-01

    This Site Specific Waste Management Instruction provides guidance for management of waste generated during the excavation and remediation of soil and debris from the 116-4 soil storage unit located at the Hanford Site in Richland, Washington. This document outlines the waste management practices that will be performed in the field to implement federal, state, and US Department of Energy requirements

  9. Development and application of bioassays for a site-specific risk assessment of contaminated soil

    NARCIS (Netherlands)

    Rila, J.-P.

    2008-01-01

    Soil risk assessment based on generic approaches is accompanied by a large number of uncertainties. In site-specific risk assessment aimed at identifying the actual effects on the ecosystem by using e.g. bioassays in soil elutriates and taking into account land-use these uncertainties can be largely

  10. Appreciating "Thirdspace": An Alternative Way of Viewing and Valuing Site-Specific Dance Performance

    Science.gov (United States)

    Munjee, Tara

    2014-01-01

    Site-specific dance performance involves the presentation of choreography in connection with a site. The context of the site combined with a viewer's personal history, beliefs, and identity impact the reading and appreciation of the performance. Although both stage and site dance performance valuing elicit multiple interpretations of artistic…

  11. Selective pressures to maintain attachment site specificity of integrative and conjugative elements.

    Directory of Open Access Journals (Sweden)

    Kayla L Menard

    Full Text Available Integrative and conjugative elements (ICEs are widespread mobile genetic elements that are usually found integrated in bacterial chromosomes. They are important agents of evolution and contribute to the acquisition of new traits, including antibiotic resistances. ICEs can excise from the chromosome and transfer to recipients by conjugation. Many ICEs are site-specific in that they integrate preferentially into a primary attachment site in the bacterial genome. Site-specific ICEs can also integrate into secondary locations, particularly if the primary site is absent. However, little is known about the consequences of integration of ICEs into alternative attachment sites or what drives the apparent maintenance and prevalence of the many ICEs that use a single attachment site. Using ICEBs1, a site-specific ICE from Bacillus subtilis that integrates into a tRNA gene, we found that integration into secondary sites was detrimental to both ICEBs1 and the host cell. Excision of ICEBs1 from secondary sites was impaired either partially or completely, limiting the spread of ICEBs1. Furthermore, induction of ICEBs1 gene expression caused a substantial drop in proliferation and cell viability within three hours. This drop was dependent on rolling circle replication of ICEBs1 that was unable to excise from the chromosome. Together, these detrimental effects provide selective pressure against the survival and dissemination of ICEs that have integrated into alternative sites and may explain the maintenance of site-specific integration for many ICEs.

  12. Validity of the Remote Food Photography Method Against Doubly Labeled Water Among Minority Preschoolers.

    Science.gov (United States)

    Nicklas, Theresa; Saab, Rabab; Islam, Noemi G; Wong, William; Butte, Nancy; Schulin, Rebecca; Liu, Yan; Apolzan, John W; Myers, Candice A; Martin, Corby K

    2017-09-01

    The aim of this study was to determine the validity of energy intake (EI) estimations made using the remote food photography method (RFPM) compared to the doubly labeled water (DLW) method in minority preschool children in a free-living environment. Seven days of food intake and spot urine samples excluding first void collections for DLW analysis were obtained on thirty-nine 3- to 5-year-old Hispanic and African American children. Using an iPhone, caregivers captured before and after pictures of each child's intake, pictures were wirelessly transmitted to trained raters who estimated portion size using existing visual estimation procedures, and energy and macronutrients were calculated. Paired t tests, mean differences, and Bland-Altman limits of agreement were performed. The mean EI was 1,191 ± 256 kcal/d using the RFPM and 1,412 ± 220 kcal/d using the DLW method, resulting in a mean underestimate of 222 kcal/d (-15.6%; P < 0.0001) that was consistent regardless of intake. The RFPM underestimated EI by -28.5% in 34 children and overestimated EI by 15.6% in 5 children. The RFPM underestimated total EI when compared to the DLW method among preschoolers. Further refinement of the RFPM is needed for assessing the EI of young children. © 2017 The Obesity Society.

  13. Comparing model-based and model-free analysis methods for QUASAR arterial spin labeling perfusion quantification.

    Science.gov (United States)

    Chappell, Michael A; Woolrich, Mark W; Petersen, Esben T; Golay, Xavier; Payne, Stephen J

    2013-05-01

    Amongst the various implementations of arterial spin labeling MRI methods for quantifying cerebral perfusion, the QUASAR method is unique. By using a combination of labeling with and without flow suppression gradients, the QUASAR method offers the separation of macrovascular and tissue signals. This permits local arterial input functions to be defined and "model-free" analysis, using numerical deconvolution, to be used. However, it remains unclear whether arterial spin labeling data are best treated using model-free or model-based analysis. This work provides a critical comparison of these two approaches for QUASAR arterial spin labeling in the healthy brain. An existing two-component (arterial and tissue) model was extended to the mixed flow suppression scheme of QUASAR to provide an optimal model-based analysis. The model-based analysis was extended to incorporate dispersion of the labeled bolus, generally regarded as the major source of discrepancy between the two analysis approaches. Model-free and model-based analyses were compared for perfusion quantification including absolute measurements, uncertainty estimation, and spatial variation in cerebral blood flow estimates. Major sources of discrepancies between model-free and model-based analysis were attributed to the effects of dispersion and the degree to which the two methods can separate macrovascular and tissue signal. Copyright © 2012 Wiley Periodicals, Inc.

  14. Points of Convergence in Music Education: The Use of Data Labels as a Strategy for Mixed Methods Integration

    Science.gov (United States)

    Fitzpatrick, Kate R.

    2016-01-01

    Although the mixing of quantitative and qualitative data is an essential component of mixed methods research, the process of integrating both types of data in meaningful ways can be challenging. The purpose of this article is to describe the use of data labels in mixed methods research as a technique for the integration of qualitative and…

  15. Spiked proteomic standard dataset for testing label-free quantitative software and statistical methods

    Directory of Open Access Journals (Sweden)

    Claire Ramus

    2016-03-01

    Full Text Available This data article describes a controlled, spiked proteomic dataset for which the “ground truth” of variant proteins is known. It is based on the LC-MS analysis of samples composed of a fixed background of yeast lysate and different spiked amounts of the UPS1 mixture of 48 recombinant proteins. It can be used to objectively evaluate bioinformatic pipelines for label-free quantitative analysis, and their ability to detect variant proteins with good sensitivity and low false discovery rate in large-scale proteomic studies. More specifically, it can be useful for tuning software tools parameters, but also testing new algorithms for label-free quantitative analysis, or for evaluation of downstream statistical methods. The raw MS files can be downloaded from ProteomeXchange with identifier http://www.ebi.ac.uk/pride/archive/projects/PXD001819. Starting from some raw files of this dataset, we also provide here some processed data obtained through various bioinformatics tools (including MaxQuant, Skyline, MFPaQ, IRMa-hEIDI and Scaffold in different workflows, to exemplify the use of such data in the context of software benchmarking, as discussed in details in the accompanying manuscript [1]. The experimental design used here for data processing takes advantage of the different spike levels introduced in the samples composing the dataset, and processed data are merged in a single file to facilitate the evaluation and illustration of software tools results for the detection of variant proteins with different absolute expression levels and fold change values.

  16. An automated pulse labelling method for structure-activity relationship studies with antibacterial oxazolidinones.

    Science.gov (United States)

    Eustice, D C; Brittelli, D R; Feldman, P A; Brown, L J; Borkowski, J J; Slee, A M

    1990-01-01

    The 3-aryl-2-oxooxazolidinones are a new class of synthetic antibacterial agents that potently inhibit protein synthesis. An automated pulse labelling method with [3H]-lysine was developed with Bacillus subtilis to obtain additional quantitative activity data for structure-activity relationship studies with the oxazolidinones. Inhibition constants were calculated after a Logit fit of the data into the formula: % of control = 100/(1 + e[-B(X - A)]), where B is the slope of the model, X is the natural log of the inhibitor concentration and A is the natural log of the inhibitor concentration required to inhibit protein synthesis by 50% (ln IC50). When substituents at the 5-methyl position of the heterocyclic ring (B-substituent) were NHCOCH3, OH or Cl, the correlation coefficient was 0.87 between the MIC and IC50 values (for all compounds with MICs less than or equal to 16 micrograms/ml). The D-isomers of DuP 721 (A-substituent = CH3CO) and DuP 105 (A-substituent = CH3SO) gave MICs of 128 micrograms/ml and IC50s of greater than or equal to 50 micrograms/ml for protein synthesis, showing that only the L-isomers were active. By MIC testing, oxazolidinones with the B-substituent of NHCOCH3 and the A-substituent of CH3CO, NO2, CH3S, CH3SO2 or (CH3)2CH had comparable antibacterial potency; however, pulse labelling analysis showed that compounds with an A-substituent of CH3CO or NO2 were more potent inhibitors of protein synthesis.

  17. Controlled, Site-Specific Functionalization of Carbon Nanotubes with Diazonium Salts

    Science.gov (United States)

    Tour, James M.

    2013-01-01

    capacities and uses. Site-specific functionalization may enable the use of nanotubes in molecular electronic applications because device functionality is critical at the cross points.

  18. Improving 1D Site Specific Velocity Profiles for the Kik-Net Network

    Science.gov (United States)

    Holt, James; Edwards, Benjamin; Pilz, Marco; Fäh, Donat; Rietbrock, Andreas

    2017-04-01

    Ground motion predication equations (GMPEs) form the cornerstone of modern seismic hazard assessments. When produced to a high standard they provide reliable estimates of ground motion/spectral acceleration for a given site and earthquake scenario. This information is crucial for engineers to optimise design and for regulators who enforce legal minimum safe design capacities. Classically, GMPEs were built upon the assumption that variability around the median model could be treated as aleatory. As understanding improved, it was noted that the propagation could be segregated into the response of the average path from the source and the response of the site. This is because the heterogeneity of the near-surface lithology is significantly different from that of the bulk path. It was then suggested that the semi-ergodic approach could be taken if the site response could be determined, moving uncertainty away from aleatory to epistemic. The determination of reliable site-specific response models is therefore becoming increasingly critical for ground motion models used in engineering practice. Today it is common practice to include proxies for site response within the scope of a GMPE, such as Vs30 or site classification, in an effort to reduce the overall uncertainty of the predication at a given site. However, these proxies are not always reliable enough to give confident ground motion estimates, due to the complexity of the near-surface. Other approaches of quantifying the response of the site include detailed numerical simulations (1/2/3D - linear, EQL, non-linear etc.). However, in order to be reliable, they require highly detailed and accurate velocity and, for non-linear analyses, material property models. It is possible to obtain this information through invasive methods, but is expensive, and not feasible for most projects. Here we propose an alternative method to derive reliable velocity profiles (and their uncertainty), calibrated using almost 20 years of

  19. LC/MS Method for the Determination of Stable Isotope Labeled Promethazine in Human Plasma

    Science.gov (United States)

    Zuwei, Wang; Boyd, Jason; Berens, Kurt L.; Putcha, Lakshmi

    2004-01-01

    Promethazine (PMZ) is taken by astronauts orally (PO), intramuscularly (IM) or rectally (PR) for space motion sickness. LC/MS method was developed with off-line solid phase extraction to measure plasma concentrations of PMZ given as stable isotope-labeled (SIL) formulations by the three different routes of administration simultaneously. Samples (0.5ml) were loaded on to Waters Oasis HLB co-polymer cartridges and eluted with 1.0 mL methanol. HPLC separation of the eluted sample was performed using an Agilent Zorbax SB-CN column (50 x 2.1 mm) at a flow rate of 0.2 mL/min for 6 min. Acetonitrile/ ammonium acetate (30 mM) in water (3:2, v/v), pH 5.6 plus or minus 0.1, was used as the mobile phase for separation. Concentrations of PMZ, PMZ-d4 and PMZ-d7 and chlorpromazine (internal standard) were determined using a Micromass ZMD single quadrupole mass spectrometer with Electrospray Ionization (ESI). ESI mass spectra were acquired in positive ion mode with selected ion monitoring of [M+ H]dot plus. The method is rapid, reproducible and the assay specific parameters are listed in a table. A novel, sensitive and specific method for the measurement of PMZ and SIL PMZ in human plasma is reported.

  20. Clustered Regularly Interspaced Short Palindromic Repeats/Cas9 Triggered Isothermal Amplification for Site-Specific Nucleic Acid Detection.

    Science.gov (United States)

    Huang, Mengqi; Zhou, Xiaoming; Wang, Huiying; Xing, Da

    2018-02-06

    A novel CRISPR/Cas9 triggered isothermal exponential amplification reaction (CAS-EXPAR) strategy based on CRISPR/Cas9 cleavage and nicking endonuclease (NEase) mediated nucleic acids amplification was developed for rapid and site-specific nucleic acid detection. CAS-EXPAR was primed by the target DNA fragment produced by cleavage of CRISPR/Cas9, and the amplification reaction performed cyclically to generate a large number of DNA replicates which were detected using a real-time fluorescence monitoring method. This strategy that combines the advantages of CRISPR/Cas9 and exponential amplification showed high specificity as well as rapid amplification kinetics. Unlike conventional nucleic acids amplification reactions, CAS-EXPAR does not require exogenous primers, which often cause target-independent amplification. Instead, primers are first generated by Cas9/sgRNA directed site-specific cleavage of target and accumulated during the reaction. It was demonstrated this strategy gave a detection limit of 0.82 amol and showed excellent specificity in discriminating single-base mismatch. Moreover, the applicability of this method to detect DNA methylation and L. monocytogenes total RNA was also verified. Therefore, CAS-EXPAR may provide a new paradigm for efficient nucleic acid amplification and hold the potential for molecular diagnostic applications.

  1. Site-specific deletions of chromosomally located DNA segments with the multimer resolution system of broad-host-range plasmid RP4

    DEFF Research Database (Denmark)

    Sternberg, Claus; Eberl, Leo; Sanchezromero, Juan M.

    1995-01-01

    The multimer resolution system (mrs) of the broad-host-range plasmid RP4 has been exploited to develop a general method that permits the precise excision of chromosomal segments in a variety of gram-negative bacteria. The procedure is based on the site-specific recombination between two directly ...

  2. Comparison of three fluorescence labeling and tracking methods of endothelial progenitor cells in laser-injured retina

    Directory of Open Access Journals (Sweden)

    Hui Shi

    2018-04-01

    Full Text Available AIM: To compare three kinds of fluorescent probes for in vitro labeling and in vivo tracking of endothelial progenitor cells (EPCs in a mouse model of laser-induced retinal injury. METHODS: EPCs were isolated from human umbilical cord blood mononuclear cells and labeled with three different fluorescent probes: 5-(and-6-carboxyfluorescein diacetate succinimidyl ester (CFSE, 1,1′-dilinoleyl-3,3,3′,3′-tetramethylindo-carbocyanine perchlorate linked acetylated low-density lipoprotein (DiI-AcLDL, and green fluorescent protein (GFP. The fluorescent intensity of EPCs was examined by confocal microscopy. Survival rate of labeled EPCs was calculated with trypan blue staining, and their adhesive capability was assessed. A mouse model of retinal injury was induced by laser, and EPCs were injected into the vitreous cavity. Frozen section and fluorescein angiography on flat-mounted retinal samples was employed to track the labeled EPCs in vivo. RESULTS: EPCs labeled with CFSE and DiI-AcLDL exhibited an intense green and red fluorescence at the beginning; the fluorescence intensity decreased gradually to 20.23% and 49.99% respectively, after 28d. On the contrary, the florescent intensity of GFP-labeled EPCs increased in a time-dependent manner. All labeled EPCs showed normal morphology and no significant change in survival and adhesive capability. In the mouse model, transplantation of EPCs showed a protective effect against retinal injury. EPCs labeled with CFSE and DiI-AcLDL were successfully tracked in mice during the development of retinal injury and repair; however, GFP-labeled EPCs were not detected in the laser-injured mouse retina. CONCLUSION: The three fluorescent markers used in this study have their own set of advantages and disadvantages. CFSE and DiI-AcLDL are suitable for short-term EPC-labeling, while GFP should be used for long-term labeling. The choice of fluorescent markers should be guided by the purpose of the study.

  3. Modified method for labeling human platelets with indium-111 oxine using albumin density-gradient separation

    International Nuclear Information System (INIS)

    Bunting, R.W.; Callahan, R.J.; Finkelstein, S.; Lees, R.S.; Strauss, H.W.

    1982-01-01

    When labeling platelets with indium-111 oxine, albumin density-gradient separation minimizes the time spent to resuspend those platelets that have been centrifuged against a hard surface. Labeling efficiency or platelet viability, as measured by platelet survival or aggregation with adenosine diphosphate, are not adversely affected

  4. Toxicity of ammonia, cadmium, and nitrobenzene to four local fishes in the Liao River, China and the derivation of site-specific water quality criteria.

    Science.gov (United States)

    Liu, Zhihong; Li, Xiaojun; Tai, Peidong; Sun, Lizong; Yuan, Honghong; Yang, Xiaonan

    2018-01-01

    Water quality criteria (WQC) are considered to be an effective management tool for protecting aquatic environments. To derive site-specific WQC for an area, local data based on local species are essential to improve the applicability of WQC derived. Due to the paucity of local fish data available for the development of site-specific WQC for the Liao River, China, four local and widespread fishes (Pseudorasbora parva, Abbottina liaoningensis, Ctenogobius giurinus, and Misgurnus anguillicaudatus) were chosen to test their sensitivities to ammonia, cadmium and nitrobenzene. These compounds are common and regularly-measured pollutants in Chinese rivers. In addition to the published data for species resident in the Liao River, site-specific WQC for the three chemicals were derived using both a log-logistic species sensitivity distribution (SSD) and the method recommended by the USEPA, in line with current best practice, which were then compared with Chinese national WQC. It was found that A. liaoningensis was the most sensitive, followed, in order, by P. parva, C. giurinus and M. anguillicaudatus was the least sensitive, and this trend was the same to all three chemicals tested. When comparing the SSD derived solely from previously-published data with that including our data on local fish, there were significant differences identified among parameters describing the SSD curves for ammonia and nitrobenzene and significant differences were detected for site-specific WQC derived for all of the three chemicals. Based on the dataset with local fish data taxa, site-specific WQC of Liao River for ammonia, cadmium, and nitrobenzene were derived to be 20.53mg/L (at a pH of 7.0 and temperature of 20°C), 3.76μg/L (at a hardness of 100mg/L CaCO 3 ), and 0.49mg/L, respectively. Using the same deriving method for each chemical, the national Chinese WQC were higher than site-specific WQC derived in this study for ammonia (national WQC of 25.16mg/L) and nitrobenzene (national WQC

  5. Development of a facile and sensitive HPLC-FLD method via fluorescence labeling for triterpenic acid bioavailability investigation.

    Science.gov (United States)

    You, Jinmao; Wu, Di; Zhao, Mei; Li, Guoliang; Gong, Peiwei; Wu, Yueyue; Guo, Yu; Chen, Guang; Zhao, Xianen; Sun, Zhiwei; Xia, Lian; Wu, Yongning

    2017-06-01

    Triterpenic acids are widely distributed in many fruits and are known for their medicinal benefits. The study of bioavailability has been an important task for a better understanding of the triterpenic acids. Although many methods based on fluorescence labeling for triterpenic acid determination have been established, these reported methods needed anhydrous conditions, which are not suitable for the convenient study of triterpenic acid bioavailability. Inspired by that, a versatile method, which overcomes the difficulty of the reported methods, has been first developed in this study. The novel method using 2-[12-benzo[b]acridin-5- (12H)-yl]-acetohydrazide (BAAH) as the fluorescence labeling reagent coupled with high-performance liquid chromatography with fluorescence detection was first developed for the study of triterpenic acid bioavailability. Furthermore, the labeling conditions have been optimized in order to achieve the best fluorescence labeling yield. Under the optimal conditions, the quantitative linear range of analytes was 2-1000 ng mL -1 , and the correlation coefficients were >0.9998. The detection limits for all triterpenic acid derivatives were achieved within the range of 0.28-0.29 ng mL -1 . The proposed method was successfully applied to the study of triterpenic acid bioavailability with excellent applicability and good reproducibility. Copyright © 2016 John Wiley & Sons, Ltd.

  6. Site-specific seismic probabilistic tsunami hazard analysis: performances and potential applications

    Science.gov (United States)

    Tonini, Roberto; Volpe, Manuela; Lorito, Stefano; Selva, Jacopo; Orefice, Simone; Graziani, Laura; Brizuela, Beatriz; Smedile, Alessandra; Romano, Fabrizio; De Martini, Paolo Marco; Maramai, Alessandra; Piatanesi, Alessio; Pantosti, Daniela

    2017-04-01

    Seismic Probabilistic Tsunami Hazard Analysis (SPTHA) provides probabilities to exceed different thresholds of tsunami hazard intensity, at a specific site or region and in a given time span, for tsunamis caused by seismic sources. Results obtained by SPTHA (i.e., probabilistic hazard curves and inundation maps) represent a very important input to risk analyses and land use planning. However, the large variability of source parameters implies the definition of a huge number of potential tsunami scenarios, whose omission could lead to a biased analysis. Moreover, tsunami propagation from source to target requires the use of very expensive numerical simulations. At regional scale, the computational cost can be reduced using assumptions on the tsunami modeling (i.e., neglecting non-linear effects, using coarse topo-bathymetric meshes, empirically extrapolating maximum wave heights on the coast). On the other hand, moving to local scale, a much higher resolution is required and such assumptions drop out, since detailed inundation maps require significantly greater computational resources. In this work we apply a multi-step method to perform a site-specific SPTHA which can be summarized in the following steps: i) to perform a regional hazard assessment to account for both the aleatory and epistemic uncertainties of the seismic source, by combining the use of an event tree and an ensemble modeling technique; ii) to apply a filtering procedure which use a cluster analysis to define a significantly reduced number of representative scenarios contributing to the hazard of a specific target site; iii) to perform high resolution numerical simulations only for these representative scenarios and for a subset of near field sources placed in very shallow waters and/or whose coseismic displacements induce ground uplift or subsidence at the target. The method is applied to three target areas in the Mediterranean located around the cities of Milazzo (Italy), Thessaloniki (Greece) and

  7. Energy costs of surgery as measured by the doubly labeled water (2H218O) method

    International Nuclear Information System (INIS)

    Novick, W.M.; Nusbaum, M.; Stein, T.P.

    1988-01-01

    Energy expenditure before and after surgery was determined in seven patients by the doubly labeled water ( 2 H 2 18 O) method (DLW). The values were compared with values obtained by respiratory gas exchange by means of a metabolic measuring cart (MMC). Patients were maintained on total parenteral nutrition before and after trauma. The principal finding was an increase in the rate of CO 2 production of 11.9 +/- 5.0% after surgery. This corresponds to a 267 +/- increase in energy expenditure (p less than 0.05). No trauma-associated change in energy expenditure was found with the MMC. The correlation of preoperative values from MMC and DLW was not statistically significant (r = 0.25), nor was the correlation of MMC and the Harris-Benedict equation, but the correlation of DLW with Harris-Benedict equation was statistically significant (r = 0.73, p less than 0.05). We suggest that the discrepancy is because the DLW method measures the cumulative energy expenditure over a period, whereas the MMC gives a spot measurement

  8. Preparation of carbon 11-labelled radiopharmaceuticals by the use of HPLC method

    International Nuclear Information System (INIS)

    Berget, G.; Maziere, M.; Godot, J.M.; Sastre, J.; Prenant, C.; Comar, D.

    1982-06-01

    Various medical examinations and metabolic studies are carried out with carbon 11-labelled radiopharmaceuticals. This radioelement offers a number of advantages: it can be introduced into an organic molecule without changing its properties; the radiation dose delivered to the patient is low (T = 20 mn); since the specific activity obtained is high (0.5 to 2 Ci/μ mole) the injected masses are very small; finally, tomographic images of the distribution of the product in the body may be obtained by the use of positron cameras. However in view of the radioactivities handled on a routine basis the preparations must be carried out without manual intervention, in closed shielded hoods. Synthesis methods and special equipment have been developed. In all cases the reaction mixtures are purified by HPLC, a method chosen for its speed, efficiency, ease of automation and adaptation to any product with a suitable choice of column and eluant. The radiopharmaceuticals are obtained in injectable solution (ethanol-physiological serum, buffered physiological serum) or in a mixture of volatile solvents which are evaporated by nitrogen bubbling and finally sterilised by passage over millipore filter. About ten different radiopharmaceuticals are prepared in this way in the laboratory [fr

  9. Development of a site specific environmental radiation assessment model for Youngkwang NPPs

    Energy Technology Data Exchange (ETDEWEB)

    Chung, Yang Geun; Song, Myung Jae; Shon, Soon Hwan; Shin, Sang Woon; Lee, Gap Bock; Yang, Kyung Hwa [Korea Electric Power Research Institute, Taejon (Korea, Republic of); Kim, Seong Pyung; Kim, Jeong Kyu; Shin, Dae Yoon; Kim, Hyun Ku; Lee, Kyung Jin [Chosun University, Kwangju (Korea, Republic of)

    1996-12-31

    During the normal operation of nuclear power plants, a very small amount of radionuclide materials is released to the environment. The radionuclide releases to the environment may give rise to some additional radiation dose through a number of pathways. With a few exception, radionuclide transports are highly dependent on the environmental conditions such as topological features, atmospheric and oceanic conditions and socio-environmental characteristics, etc. The dose calculation in Korea are presently being performed using the models in US NRC Reg. Guide. However, these models which were originally released by the US NRC in 1977 are inadequate to deal with environmental movement of radionuclides in consideration of complex terrain, tidal condition of the Yellow Sea, and Youngkwang site specific conditions. KEPRI has developed the models to consider the site specific characteristics around Youngkwang NPPs, to give the realistic dose assessment, and to improve assurance and reliability in public dose calculation. (author). 41 figs., 70 refs.

  10. Recent Developments in the Site-Specific Immobilization of Proteins onto Solid Supports

    Energy Technology Data Exchange (ETDEWEB)

    Camarero, J A

    2007-02-21

    Immobilization of proteins onto surfaces is of great importance in numerous applications, including protein analysis, drug screening, and medical diagnostics, among others. The success of all these technologies relies on the immobilization technique employed to attach a protein to the corresponding surface. Non-specific physical adsorption or chemical cross-linking with appropriate surfaces results in the immobilization of the protein in random orientations. Site-specific covalent attachment, on the other hand, leads to molecules being arranged in a definite, orderly fashion and allows the use of spacers and linkers to help minimize steric hindrances between the protein and the surface. The present work reviews the latest chemical and biochemical developments for the site-specific covalent attachment of proteins onto solid supports.

  11. A Molecular Toolbox to Engineer Site-Specific DNA Replication Perturbation.

    Science.gov (United States)

    Larsen, Nicolai B; Hickson, Ian D; Mankouri, Hocine W

    2018-01-01

    Site-specific arrest of DNA replication is a useful tool for analyzing cellular responses to DNA replication perturbation. The E. coli Tus-Ter replication barrier can be reconstituted in eukaryotic cells as a system to engineer an unscheduled collision between a replication fork and an "alien" impediment to DNA replication. To further develop this system as a versatile tool, we describe a set of reagents and a detailed protocol that can be used to engineer Tus-Ter barriers into any locus in the budding yeast genome. Because the Tus-Ter complex is a bipartite system with intrinsic DNA replication-blocking activity, the reagents and protocols developed and validated in yeast could also be optimized to engineer site-specific replication fork barriers into other eukaryotic cell types.

  12. A site-specific slurry application technique on grassland and on arable crops.

    Science.gov (United States)

    Schellberg, Jürgen; Lock, Reiner

    2009-01-01

    There is evidence that unequal slurry application on agricultural land contributes to N losses to the environment. Heterogeneity within fields demands adequate response by means of variable rate application. A technique is presented which allows site-specific application of slurry on grassland and arable land based on pre-defined application maps. The system contains a valve controlling flow rate by an on-board PC. During operation, flow rate is measured and scaled against set point values given in the application map together with the geographic position of the site. The systems worked sufficiently precise at a flow rate between 0 and 25 l s(-1) and an offset of actual slurry flow from set point values between 0.33 and 0.67 l s(-1). Long-term experimentation is required to test if site-specific application de facto reduces N surplus within fields and so significantly contributes to the unloading of N in agricultural areas.

  13. Tus-Ter as a tool to study site-specific DNA replication perturbation in eukaryotes

    DEFF Research Database (Denmark)

    Larsen, Nicolai B; Hickson, Ian D; Mankouri, Hocine W

    2014-01-01

    The high-affinity binding of the Tus protein to specific 21-bp sequences, called Ter, causes site-specific, and polar, DNA replication fork arrest in E coli. The Tus-Ter complex serves to coordinate DNA replication with chromosome segregation in this organism. A number of recent and ongoing studies...... have demonstrated that Tus-Ter can be used as a heterologous tool to generate site-specific perturbation of DNA replication when reconstituted in eukaryotes. Here, we review these recent findings and explore the molecular mechanism by which Tus-Ter mediates replication fork (RF) arrest in the budding...... yeast, S. cerevisiae. We propose that Tus-Ter is a versatile, genetically tractable, and regulatable RF blocking system that can be utilized for disrupting DNA replication in a diverse range of host cells....

  14. Tus-Ter as a tool to study site-specific DNA replication perturbation in eukaryotes.

    Science.gov (United States)

    Larsen, Nicolai B; Hickson, Ian D; Mankouri, Hocine W

    2014-01-01

    The high-affinity binding of the Tus protein to specific 21-bp sequences, called Ter, causes site-specific, and polar, DNA replication fork arrest in E coli. The Tus-Ter complex serves to coordinate DNA replication with chromosome segregation in this organism. A number of recent and ongoing studies have demonstrated that Tus-Ter can be used as a heterologous tool to generate site-specific perturbation of DNA replication when reconstituted in eukaryotes. Here, we review these recent findings and explore the molecular mechanism by which Tus-Ter mediates replication fork (RF) arrest in the budding yeast, S. cerevisiae. We propose that Tus-Ter is a versatile, genetically tractable, and regulatable RF blocking system that can be utilized for disrupting DNA replication in a diverse range of host cells.

  15. Proposed Site-Specific Response Spectra for Surabaya-Madura Bridge

    Directory of Open Access Journals (Sweden)

    Dyah Kusumastuti

    2008-01-01

    Full Text Available This paper presents a site-specific seismic hazard study to determine the recommended seismic design criteria for Suramadu Bridge. The study is performed using probabilistic seismic hazard approach to determine maximum acceleration and response spectra at bedrock and followed by local site effect analysis to determine maximum acceleration and response spectra at ground surface. The probabilistic seismic hazard analysis (PSHA is carried out using 3-dimension (3-D seismic source models (fault source model. Two hazard levels are analysed to represent 150 and 3,300 years return period of ground motion around site location. The local site effect analysis is performed using 1-dimension (1-D shear wave propagation theory to obtain peak ground acceleration and response spectra at ground surface. Finally, the site-specific surface response spectra with 5 percent damping are developed based on the mean plus one standard deviation concept from the result of local site effect analysis.

  16. Site-specifically modified oligodeoxyribonucleotides as templates for Escherichia coli DNA polymerase I

    International Nuclear Information System (INIS)

    O'Connor, D.; Stoehrer, G.

    1985-01-01

    Oligodeoxyribonucleotides with site-specific modifications have been used as substrates for Escherichia coli DNA polymerase I holoenzyme and Klenow fragment. Modifications included the bulky guanine-8-aminofluorene adduct and a guanine oxidation product resembling the product of photosensitized DNA oxidation. By a combination of primers and nick-mers, conditions of single-strand-directed DNA synthesis and nick-translation could be created. The results show that the polymerase can bypass both types of lesions. Bypass occurs on a single-stranded template but is facilitated on a nicked, double-stranded template. Only purines, with guanine more favored than adenine, are incorporated across both lesions. The results indicate that site-specifically modified oligonucleotides can be sensitive probes for the action of polymerases on damaged templates. They also suggest a function for polymerase I, in its nick-translation capacity, during DNA repair and mutagenesis

  17. Site-specific growth of Au particles on ZnO nanopyramids under ultraviolet illumination

    KAUST Repository

    Yao, Kexin

    2011-01-01

    In this work, wurtzite ZnO nanocrystals with unique "pyramid" morphology were firstly prepared via solvothermal synthesis. It was determined that the ZnO nanopyramids are grown along the polar c-axis with the vertexes pointing to the [001] direction. When the mixture of ZnO nanopyramids and Au precursor (HAuCl4) was exposed to ultraviolet (UV) illumination, Au particles were site-specifically formed on the vertexes of ZnO nanopyramids. The obtained Au/ZnO nanocomposite showed significantly enhanced photocatalytic activity as compared to the bare ZnO nanopyramids. First-principles based calculations well explained the formation of ZnO nanopyramids as well as the site-specific growth of Au, and revealed that during the photocatalysis process the Au particles can accommodate photoelectrons and thus facilitate the charge separation. © 2011 The Royal Society of Chemistry.

  18. Site-Specific Integration of Exogenous Genes Using Genome Editing Technologies in Zebrafish

    Directory of Open Access Journals (Sweden)

    Atsuo Kawahara

    2016-05-01

    Full Text Available The zebrafish (Danio rerio is an ideal vertebrate model to investigate the developmental molecular mechanism of organogenesis and regeneration. Recent innovation in genome editing technologies, such as zinc finger nucleases (ZFNs, transcription activator-like effector nucleases (TALENs and the clustered regularly interspaced short palindromic repeats (CRISPR/CRISPR associated protein 9 (Cas9 system, have allowed researchers to generate diverse genomic modifications in whole animals and in cultured cells. The CRISPR/Cas9 and TALEN techniques frequently induce DNA double-strand breaks (DSBs at the targeted gene, resulting in frameshift-mediated gene disruption. As a useful application of genome editing technology, several groups have recently reported efficient site-specific integration of exogenous genes into targeted genomic loci. In this review, we provide an overview of TALEN- and CRISPR/Cas9-mediated site-specific integration of exogenous genes in zebrafish.

  19. A Molecular Toolbox to Engineer Site-Specific DNA Replication Perturbation

    DEFF Research Database (Denmark)

    Larsen, Nicolai B; Hickson, Ian D; Mankouri, Hocine W

    2018-01-01

    " impediment to DNA replication. To further develop this system as a versatile tool, we describe a set of reagents and a detailed protocol that can be used to engineer Tus-Ter barriers into any locus in the budding yeast genome. Because the Tus-Ter complex is a bipartite system with intrinsic DNA replication......Site-specific arrest of DNA replication is a useful tool for analyzing cellular responses to DNA replication perturbation. The E. coli Tus-Ter replication barrier can be reconstituted in eukaryotic cells as a system to engineer an unscheduled collision between a replication fork and an "alien......-blocking activity, the reagents and protocols developed and validated in yeast could also be optimized to engineer site-specific replication fork barriers into other eukaryotic cell types....

  20. Long-range chromosome organization in E. coli: a site-specific system isolates the Ter macrodomain.

    Science.gov (United States)

    Thiel, Axel; Valens, Michèle; Vallet-Gely, Isabelle; Espéli, Olivier; Boccard, Frédéric

    2012-01-01

    The organization of the Escherichia coli chromosome into a ring composed of four macrodomains and two less-structured regions influences the segregation of sister chromatids and the mobility of chromosomal DNA. The structuring of the terminus region (Ter) into a macrodomain relies on the interaction of the protein MatP with a 13-bp target called matS repeated 23 times in the 800-kb-long domain. Here, by using a new method that allows the transposition of any chromosomal segment at a defined position on the genetic map, we reveal a site-specific system that restricts to the Ter region a constraining process that reduces DNA mobility and delays loci segregation. Remarkably, the constraining process is regulated during the cell cycle and occurs only when the Ter MD is associated with the division machinery at mid-cell. The change of DNA properties does not rely on the presence of a trans-acting mechanism but rather involves a cis-effect acting at a long distance from the Ter region. Two specific 12-bp sequences located in the flanking Left and Right macrodomains and a newly identified protein designated YfbV conserved with MatP through evolution are required to impede the spreading of the constraining process to the rest of the chromosome. Our results unravel a site-specific system required to restrict to the Ter region the consequences of anchoring the Ter MD to the division machinery.

  1. Site-specific and multielement approach to the determination of liquid-vapor isotope fractionation parameters. The case of alcohols

    International Nuclear Information System (INIS)

    Moussa, I.; Naulet, N.; Martin, M.L.; Martin, G.J.

    1990-01-01

    Isotope fractionation phenomena occurring at the natural abundance level in the course of liquid-vapor transformation have been investigated by using the SNIF-NMR method (site-specific natural isotope fractionation studied by NMR) which has a unique capability of providing simultaneous access to fractionation parameters associated with different molecular isotopomers. This new approach has been combined with the determination of overall carbon and hydrogen fractionation effects by isotope ratio mass spectrometry (IRMS). The results of distillation and evaporation experiments of alcohols performed in technical conditions of practical interest have been analyzed according to the Rayleigh-type model. In order to check the performance of the column, unit fractionation factors were measured beforehand for water and for the hydroxylic sites of methanol and ethanol for which liquid-vapor equilibrium constants were already known. Inverse isotope effects are determined in distillation experiments for the overall carbon isotope ratio and for the site-specific hydrogen isotope ratios associated with the methyl and methylene sites of methanol and ethanol. In contrast, normal isotope effects are produced by distillation for the hydroxylic sites and by evaporation for all the isotopic ratios

  2. New miRNA labeling method for bead-based quantification

    Directory of Open Access Journals (Sweden)

    Lanfranchi Gerolamo

    2010-06-01

    Full Text Available Abstract Background microRNAs (miRNAs are small single-stranded non-coding RNAs that act as crucial regulators of gene expression. Different methods have been developed for miRNA expression profiling in order to better understand gene regulation in normal and pathological conditions. miRNAs expression values obtained from large scale methodologies such as microarrays still need a validation step with alternative technologies. Results Here we have applied with an innovative approach, the Luminex® xMAP™ technology validate expression data of differentially expressed miRNAs obtained from high throughput arrays. We have developed a novel labeling system of small RNA molecules (below 200 nt, optimizing the sensitive cloning method for miRNAs, termed miRNA amplification profiling (mRAP. The Luminex expression patterns of three miRNAs (miR-23a, miR-27a and miR-199a in seven different cell lines have been validated by TaqMan miRNA assay. In all cases, bead-based meas were confirmed by the data obtained by TaqMan and microarray technologies. Conclusions We demonstrate that the measure of individual miRNA by the bead-based method is feasible, high speed, sensitive and low cost. The Luminex® xMAP™ technology also provides flexibility, since the central reaction can be scaled up with additional miRNA capturing beads, allowing validation of many differentially expressed miRNAs obtained from microarrays in a single experiment. We propose this technology as an alternative method to qRT-PCR for validating miRNAs expression data obtained with high-throughput technologies.

  3. Site-Specific Waste Management Instruction - 100-DR-1 Group 2 Sites

    International Nuclear Information System (INIS)

    Jackson, R.W.

    1998-01-01

    This site-specific waste management instruction (SSWMI) provides guidance for the management of wastes that may be generated during the excavation and remediation of the 100-DR-1 Group 2 sites. The management of waste generated as a result of these activities will be as directed in this SSWMI. This SSWMI will be revised to incorporate guidance for management of wastes encountered that are not addressed in this SSWMI

  4. Using a site-specific technical error to establish training responsiveness: a preliminary explorative study.

    Science.gov (United States)

    Weatherwax, Ryan M; Harris, Nigel K; Kilding, Andrew E; Dalleck, Lance C

    2018-01-01

    Even though cardiorespiratory fitness (CRF) training elicits numerous health benefits, not all individuals have positive training responses following a structured CRF intervention. It has been suggested that the technical error (TE), a combination of biological variability and measurement error, should be used to establish specific training responsiveness criteria to gain further insight on the effectiveness of the training program. To date, most training interventions use an absolute change or a TE from previous findings, which do not take into consideration the training site and equipment used to establish training outcomes or the specific cohort being evaluated. The purpose of this investigation was to retrospectively analyze training responsiveness of two CRF training interventions using two common criteria and a site-specific TE. Sixteen men and women completed two maximal graded exercise tests and verification bouts to identify maximal oxygen consumption (VO 2 max) and establish a site-specific TE. The TE was then used to retrospectively analyze training responsiveness in comparison to commonly used criteria: percent change of >0% and >+5.6% in VO 2 max. The TE was found to be 7.7% for relative VO 2 max. χ 2 testing showed significant differences in all training criteria for each intervention and pooled data from both interventions, except between %Δ >0 and %Δ >+7.7% in one of the investigations. Training nonresponsiveness ranged from 11.5% to 34.6%. Findings from the present study support the utility of site-specific TE criterion to quantify training responsiveness. A similar methodology of establishing a site-specific and even cohort specific TE should be considered to establish when true cardiorespiratory training adaptations occur.

  5. Development of the NUMO pre-selection, site-specific safety case

    International Nuclear Information System (INIS)

    Fujiyama, Tetsuo; Suzuki, Satoru; Deguchi, Akira; Umeki, Hiroyuki

    2016-01-01

    Key conclusions: ◆ “The NUMO pre-selection, site-specific safety case” provides the basic structure for subsequent safety cases that will be applied to any selected site, emphasising practical approaches and methodology which will be applicable for the conditions/constraints during an actual siting process. ◆ The preliminary results of the design and safety assessment would underpin the feasibility and safety of geological disposal in Japan.

  6. Site-specific waste management instruction for the 100-KR-4 Operable Unit drilling

    International Nuclear Information System (INIS)

    Hadley, J.T.

    1996-07-01

    This site-specific waste management instruction provides guidance for the management of waste generated as a result of groundwater well installations in the 100-KR-4 Operable Unit (OU). The well installations are necessary to implement the Remedial Action (RA) option (pump-and-treat using ion exchange) to prevent discharge of hexavalent chromium at levels above those considered protective of aquatic life in the Columbia River and riverbed sediments

  7. Site-specific waste management instruction for the 100-KR-4 Operable Unit drilling. Revision 1

    International Nuclear Information System (INIS)

    Hadley, J.T.

    1996-08-01

    This site-specific waste management instruction provides guidance for the management of waste generated as a result of groundwater well installations in the 100-KR-4 Operable Unit (OU). The well installations are necessary to implement the Remedial Action (RA) option (pump-and-treat using ion exchange) to prevent discharge of hexavalent chromium at levels above those considered protective of aquatic life in the Columbia River and riverbed sediments

  8. 77 FR 74838 - Environmental Management Site-Specific Advisory Board, Northern New Mexico

    Science.gov (United States)

    2012-12-18

    ...This notice announces a combined meeting of the Environmental Monitoring, Surveillance and Remediation Committee and Waste Management Committee of the Environmental Management Site-Specific Advisory Board (EM SSAB), Northern New Mexico (known locally as the Northern New Mexico Citizens' Advisory Board [NNMCAB]). The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  9. 78 FR 10612 - Environmental Management Site-Specific Advisory Board, Northern New Mexico

    Science.gov (United States)

    2013-02-14

    ...This notice announces a combined meeting of the Environmental Monitoring, Surveillance and Remediation Committee and Waste Management Committee of the Environmental Management Site-Specific Advisory Board (EM SSAB), Northern New Mexico (known locally as the Northern New Mexico Citizens' Advisory Board [NNMCAB]). The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  10. 77 FR 64800 - Environmental Management Site-Specific Advisory Board, Northern New Mexico

    Science.gov (United States)

    2012-10-23

    ...This notice announces a combined meeting of the Environmental Monitoring, Surveillance and Remediation Committee and Waste Management Committee of the Environmental Management Site-Specific Advisory Board (EM SSAB), Northern New Mexico (known locally as the Northern New Mexico Citizens' Advisory Board [NNMCAB]). The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  11. 78 FR 4140 - Environmental Management Site-Specific Advisory Board, Northern New Mexico

    Science.gov (United States)

    2013-01-18

    ...This notice announces a combined meeting of the Environmental Monitoring, Surveillance and Remediation Committee and Waste Management Committee of the Environmental Management Site-Specific Advisory Board (EM SSAB), Northern New Mexico (known locally as the Northern New Mexico Citizens' Advisory Board [NNMCAB]). The Federal Advisory Committee Act (Pub. L. 92-463, 86 Stat. 770) requires that public notice of this meeting be announced in the Federal Register.

  12. Site-specific cancer risk in the Baltic cohort of Chernobyl cleanup workers, 1986-2007.

    Science.gov (United States)

    Rahu, Kaja; Hakulinen, Timo; Smailyte, Giedre; Stengrevics, Aivars; Auvinen, Anssi; Inskip, Peter D; Boice, John D; Rahu, Mati

    2013-09-01

    To assess site-specific cancer risk in the Baltic cohort of Chernobyl cleanup workers, 1986-2007. The Baltic cohort includes 17,040 men from Estonia, Latvia and Lithuania who participated in the environmental cleanup after the accident at the Chernobyl Nuclear Power Station in 1986-1991 and who were followed up for cancer incidence until the end of 2007. Cancer cases diagnosed in the cohort and in the male population of each country were identified from the respective national cancer registers. The proportional incidence ratio (PIR) with 95% confidence interval (CI) was used to estimate the site-specific cancer risk in the cohort. For comparison and as it was possible, the site-specific standardised incidence ratio (SIR) was calculated for the Estonian sub-cohort, which was not feasible for the other countries. Overall, 756 cancer cases were reported during 1986-2007. A higher proportion of thyroid cancers in relation to the male population was found (PIR=2.76; 95%CI 1.63-4.36), especially among those who started their mission shortly after the accident, in April-May 1986 (PIR=6.38; 95%CI 2.34-13.89). Also, an excess of oesophageal cancers was noted (PIR=1.52; 95% CI 1.06-2.11). No increased PIRs for leukaemia or radiation-related cancer sites combined were observed. PIRs and SIRs for the Estonian sub-cohort demonstrated the same site-specific cancer risk pattern. Consistent evidence of an increase in radiation-related cancers in the Baltic cohort was not observed with the possible exception of thyroid cancer, where conclusions are hampered by known medical examination including thyroid screening among cleanup workers. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Determining site-specific background level with geostatistics for remediation of heavy metals in neighborhood soils

    OpenAIRE

    Tammy M. Milillo; Gaurav Sinha; Joseph A. Gardella Jr.

    2017-01-01

    The choice of a relevant, uncontaminated site for the determination of site-specific background concentrations for pollutants is critical for planning remediation of a contaminated site. The guidelines used to arrive at concentration levels vary from state to state, complicating this process. The residential neighborhood of Hickory Woods in Buffalo, NY is an area where heavy metal concentrations and spatial distributions were measured to plan remediation. A novel geostatistics based decision ...

  14. 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

  15. Micro-Droplet Detection Method for Measuring the Concentration of Alkaline Phosphatase-Labeled Nanoparticles in Fluorescence Microscopy

    Directory of Open Access Journals (Sweden)

    Rufeng Li

    2017-11-01

    Full Text Available This paper developed and evaluated a quantitative image analysis method to measure the concentration of the nanoparticles on which alkaline phosphatase (AP was immobilized. These AP-labeled nanoparticles are widely used as signal markers for tagging biomolecules at nanometer and sub-nanometer scales. The AP-labeled nanoparticle concentration measurement can then be directly used to quantitatively analyze the biomolecular concentration. Micro-droplets are mono-dispersed micro-reactors that can be used to encapsulate and detect AP-labeled nanoparticles. Micro-droplets include both empty micro-droplets and fluorescent micro-droplets, while fluorescent micro-droplets are generated from the fluorescence reaction between the APs adhering to a single nanoparticle and corresponding fluorogenic substrates within droplets. By detecting micro-droplets and calculating the proportion of fluorescent micro-droplets to the overall micro-droplets, we can calculate the AP-labeled nanoparticle concentration. The proposed micro-droplet detection method includes the following steps: (1 Gaussian filtering to remove the noise of overall fluorescent targets, (2 a contrast-limited, adaptive histogram equalization processing to enhance the contrast of weakly luminescent micro-droplets, (3 an red maximizing inter-class variance thresholding method (OTSU to segment the enhanced image for getting the binary map of the overall micro-droplets, (4 a circular Hough transform (CHT method to detect overall micro-droplets and (5 an intensity-mean-based thresholding segmentation method to extract the fluorescent micro-droplets. The experimental results of fluorescent micro-droplet images show that the average accuracy of our micro-droplet detection method is 0.9586; the average true positive rate is 0.9502; and the average false positive rate is 0.0073. The detection method can be successfully applied to measure AP-labeled nanoparticle concentration in fluorescence microscopy.

  16. Micro-Droplet Detection Method for Measuring the Concentration of Alkaline Phosphatase-Labeled Nanoparticles in Fluorescence Microscopy.

    Science.gov (United States)

    Li, Rufeng; Wang, Yibei; Xu, Hong; Fei, Baowei; Qin, Binjie

    2017-11-21

    This paper developed and evaluated a quantitative image analysis method to measure the concentration of the nanoparticles on which alkaline phosphatase (AP) was immobilized. These AP-labeled nanoparticles are widely used as signal markers for tagging biomolecules at nanometer and sub-nanometer scales. The AP-labeled nanoparticle concentration measurement can then be directly used to quantitatively analyze the biomolecular concentration. Micro-droplets are mono-dispersed micro-reactors that can be used to encapsulate and detect AP-labeled nanoparticles. Micro-droplets include both empty micro-droplets and fluorescent micro-droplets, while fluorescent micro-droplets are generated from the fluorescence reaction between the APs adhering to a single nanoparticle and corresponding fluorogenic substrates within droplets. By detecting micro-droplets and calculating the proportion of fluorescent micro-droplets to the overall micro-droplets, we can calculate the AP-labeled nanoparticle concentration. The proposed micro-droplet detection method includes the following steps: (1) Gaussian filtering to remove the noise of overall fluorescent targets, (2) a contrast-limited, adaptive histogram equalization processing to enhance the contrast of weakly luminescent micro-droplets, (3) an red maximizing inter-class variance thresholding method (OTSU) to segment the enhanced image for getting the binary map of the overall micro-droplets, (4) a circular Hough transform (CHT) method to detect overall micro-droplets and (5) an intensity-mean-based thresholding segmentation method to extract the fluorescent micro-droplets. The experimental results of fluorescent micro-droplet images show that the average accuracy of our micro-droplet detection method is 0.9586; the average true positive rate is 0.9502; and the average false positive rate is 0.0073. The detection method can be successfully applied to measure AP-labeled nanoparticle concentration in fluorescence microscopy.

  17. Non-site-specific allosteric effect of oxygen on human hemoglobin under high oxygen partial pressure.

    Science.gov (United States)

    Takayanagi, Masayoshi; Kurisaki, Ikuo; Nagaoka, Masataka

    2014-04-08

    Protein allostery is essential for vital activities. Allosteric regulation of human hemoglobin (HbA) with two quaternary states T and R has been a paradigm of allosteric structural regulation of proteins. It is widely accepted that oxygen molecules (O2) act as a "site-specific" homotropic effector, or the successive O2 binding to the heme brings about the quaternary regulation. However, here we show that the site-specific allosteric effect is not necessarily only a unique mechanism of O2 allostery. Our simulation results revealed that the solution environment of high O2 partial pressure enhances the quaternary change from T to R without binding to the heme, suggesting an additional "non-site-specific" allosteric effect of O2. The latter effect should play a complementary role in the quaternary change by affecting the intersubunit contacts. This analysis must become a milestone in comprehensive understanding of the allosteric regulation of HbA from the molecular point of view.

  18. Comparisons of CAP88PC version 2.0 default parameters to site specific inputs

    International Nuclear Information System (INIS)

    Lehto, M. A.; Courtney, J. C.; Charter, N.; Egan, T.

    2000-01-01

    The effects of varying the input for the CAP88PC Version 2.0 program on the total effective dose equivalents (TEDEs) were determined for hypothetical releases from the Hot Fuel Examination Facility (HFEF) located at the Argonne National Laboratory site on the Idaho National Engineering and Environmental Laboratory (INEEL). Values for site specific meteorological conditions and agricultural production parameters were determined for the 80 km radius surrounding the HFEF. Four nuclides, 3 H, 85 Kr, 129 I, and 137 Cs (with its short lived progeny, 137m Ba) were selected for this study; these are the radioactive materials most likely to be released from HFEF under normal or abnormal operating conditions. Use of site specific meteorological parameters of annual precipitation, average temperature, and the height of the inversion layer decreased the TEDE from 137 Cs- 137m Ba up to 36%; reductions for other nuclides were less than 3%. Use of the site specific agricultural parameters reduced TEDE values between 7% and 49%, depending on the nuclide. Reductions are associated with decreased committed effective dose equivalents (CEDEs) from the ingestion pathway. This is not surprising since the HFEF is located well within the INEEL exclusion area, and the surrounding area closest to the release point is a high desert with limited agricultural diversity. Livestock and milk production are important in some counties at distances greater than 30 km from the HFEF

  19. Site-specific parameter values for the Nuclear Regulatory Commission's food pathway dose model

    International Nuclear Information System (INIS)

    Hamby, D.M.

    1992-01-01

    Routine operations at the Savannah River Site (SRS) in Western South Carolina result in radionuclide releases to the atmosphere and to the Savannah River. The resulting radiation doses to the off-site maximum individual and the off-site population within 80 km of the SRS are estimated on a yearly basis. These estimates are currently generated using dose models prescribed for the commercial nuclear power industry by the Nuclear Regulatory Commission (NRC). The NRC provides default values for dose-model parameters for facilities without resources to develop site-specific values. A survey of land- and water-use characteristics for the Savannah River area has been conducted to determine site-specific values for water recreation, consumption, and agricultural parameters used in the NRC Regulatory Guide 1.109 (1977) dosimetric models. These site parameters include local characteristics of meat, milk, and vegetable production; recreational and commercial activities on the Savannah River; and meat, milk, vegetable, and seafood consumption rates. This paper describes how parameter data were obtained at the Savannah River Site and the impacts of such data on off-site dose. Dose estimates using site-specific parameter values are compared to estimates using the NRC default values

  20. Site-Specific Multilevel Modeling of Potato Response to Nitrogen Fertilization

    Directory of Open Access Journals (Sweden)

    Serge-Étienne Parent

    2017-12-01

    Full Text Available Technologies of precision agriculture, digital soil maps, and meteorological stations provide a minimum data set to guide precision farming operations. However, determining optimal nutrient requirements for potato (Solanum tuberosum L. crops at subfield scale remains a challenge given specific climatic, edaphic, and managerial conditions. Multilevel modeling can generalize yield response to fertilizer additions using data easily accessible to growers. Our objective was to elaborate a multilevel N fertilizer response model for potato crops using the Mitscherlich equation and a core data set of 93 N fertilizer trials conducted in Québec, Canada. Daily climatic data were collected at 10 × 10 km resolution. Soils were characterized by organic matter content, pH, and texture in the arable layer, and by texture and tools of pedometrics across a gleization-podzolization continuum in subsoil layers. There were five categories of preceding crops and five cultivar maturity orders. The three Mitscherlich parameters (Asymptote, Rate, and Environment were most often site-specific. Sensitivity analysis showed that optimum N dosage increased with non-leguminous high-residue preceding crops, coarser soils, podzolization, drier climatic condition, and late cultivar maturity. The inferential model could guide site-specific N fertilization using an accessible minimum data set to support fertilization decisions. As decision-support system, the model could also provide a range of optimum N doses across a large spectrum of site-specific conditions including climate change.